i
TEMPORAL VARIATION IN DINOFLAGELLATE SPECIES COMPOSITION AT FISH CAGES FARM AREA ALONG
SANTUBONG RIVER
Nurul Nadia Binti Rajami
QL 368 D6 Bachelor of Science with Honours N974 (Aquatic Resource Science and Management) 2012 2012
i Pusat Khidmat Maklumat Akademik VNlVERSm MALAYSIA SARAWAK
TEMPORAL VARIATION IN DINOFLAGELLATE SPECIES COMPOSITION IN FISH CAGES FARM AREA ALONG SANTUBONG RIVER
PKHIDMAT MAKLUMAT AKADEMIK
111111111 fonr IIII 11111 1000235641
NURUL NADIA BINTI RAJAMI
Report Submitted in partial fulfilment for the degree of Bachelor of Science with Honours (Aquatic Resource Science and Management)
t Faculty of Resource Science and Technology
University Malaysia Sarawak 2012
I
I
bull
DECLARATION
I hereby declare that the work in this project is my own except for qoutations and summaries
which have been duly acknowledged No portion of the work referred to in this dissertation
has been summitted in support of an application for another degree qualification of this or any
other university or institution of higher learning
~
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Programme
Faculty of Resource Science and Technology
University Malaysia Sarawak
bull ~ f i
ACKNOWLEDGEMENT
Alhamdulillah Thanks to ALLAH SWT for His blessings that I finally completed my final
year project This final year project report was prepared for the partial fulfilment for the
degree of Bachelor of Science in Aquatic Resource Science and Management
I would like to express my deepest thanks to many individuals who assisted me in the
completion of this project especially to my supervisor Assoc Prof Dr Othman Bojo for his
advice motivation guidance encouragement and critics My appreciation also goes to En
Zaidi Ibrahim and other lab assistants who had helped me a lot during my sampling trips I
also want to thanks the lecturers and staffs of Faculty of Resource Science and Technology
for their cooperation during the completion of the final year project They that had given me
valuable information suggestions and guidance in the compilation and preparation this final
year project report To all my course mates thanks for the cooperation helpful suggestion and
full of support for the report completion from the beginning till the end
Finally to my beloved parents Mr Rajami bin Bujang and Mrs Sarimah binti Fadzil for being
patient thoughtful and supportive Last but not least to any individual that involved directly
and indirectly in the making of this final year project report
Thank you all
I
bull r Pusat Khidmat MakJumat Akademik UNlVERSm MALAYSIA SARAWAK
Table of Contents
ACKNOWLEDGEMENT I
T ABLE OF CONTENTS II
LIST OF ABBREVIATIONS III
LIST OF TABLES IV
LIST OF FIGURES V
ABSTRACT VI
10 INTRODUCTION
20 LITERATURE REVIEW
21 Taxonomy of Dinoflagellates 3 211 Dinoflagellates morphology 3
22 Distribution of Dinoflagellates 5 23 Ecology and Growth of Dinoflagellates 6 24 Harmful Algal Blooms (HABs) 6
241 Impacts of HABs 8
30 MATERIALS AND METHODS
31 Study Areas 9 32 Phytoplankton Sampling and Water Quality Collection 11 33 Phytoplankton identification and enumeration 12 34 Statistical Data Analysis 12
40 RESULTS
41 Qualitative Phytoplankton Composition 13 42 Dinoflagellate Assemblages 14
421 Dinoflagellates Taxonomic description 24 )43 Diatom Assemblages 30
431 Diatom Taxonomic Description 31 44 Hydro-physical Characteristics
441 pH 32 442 Temperature 33 443 Salinity 34 444 Dissolved Oxygen (DO) 35 445 Turbidity 36
50 DISCUSSION 37
60 CONCLUSIONS 43
REFERENCES 45
APPENDIX 49
I i
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LIST OF ABBREVIATIONS
~m Micro Metre
ANOVA Analysis Of Variance
APHA American Public Health Association
CFP Ciguatera Fish Poisoning
Chl-a Chlorophyll A
DNA Deoxyribonucleic Acid
DO Dissolved Oxygen
DSP Diarrhetic Shellfish Poisoning
DTXs Dinophysis Toxins
GPS Global Positioning System
HABs Harmful Algal Blooms
L Litre
Mgll Milligram Per Litre
NSP Neurotoxic Shellfish Poisoning
NTU Nephelometric Turbidity Units
PbTxs Brevetoxins
PSP Paralytic Shellfish Poisoning
PSU Practical Salinity Units
PTXs Pectenotoxins
SPSS Statistical Package For Social Science
STXs Saxitoxin
YTXs Yessotoxins
L--_ J--__ _____ _
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
List Of Tables
Sampling Sites (Station) Along The Santubong River
Taxonomic List And Occurrence Of Dinoflagellates In Santubong River
Other Phytoplankton Identified In Samples Along Santubong River
ANOVA Table For Physico-Chemical Parameters
Multiple Comparison For Physic-Chemical Parameters
The Depth Of Water Column And Its Transparency
Pages
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50
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List Of Figures
Figure I Basic Anatomy of Thecate Dinokont Dinoflagellates
Figure 2 Maps of Santubong River
Figure 3 Percentage Distribution of Phytoplanktons
Figure 4 Number of Dinoflagellates Species Identified in Monthly Sampling
Figure 5 Number oflndividual per ml of Dinoflagellates along Santubong River
Figure 6a Representative Dinoflagellates
Figure 6b Representative Dinoflagellates
Figure 7a Representative Centric Diatoms
Figure 7b Representative Pennate Diatoms
Figure 8 Percentage Abundance of Diatoms along Santubong River
Figure 9 The Mean of Ph Value in Three Sampling Stations from December 2011 to April 2012
Figure 10 The Mean Value of Temperature in Three Sampling Stations from December 2011 to April 2012
Figure 11 The Mean Value of Salinity in Three Sampling Stations from December 2011 to April 2012
Figure 12 The Mean Value of Dissolved Oxygen CDO) in Three Sampling Stations from December 2011 to April 2012
Figure 13 The Mean Value of Turbidity in Three Sampling Stations from December 2011 to April 2012
Pages
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Temporal Variation of Dinoflagellates Species Composition at Fish Cages Area along Santubong River
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Faculty of Resources Science and Technology
University Malaysia Sarawak
ABSTRACT
Dinoflagellates were identified and counted from preserved net haul samples collected from three different stations along Santubong River The samples were collected monthly from December 20 II to Apri l 20 12 Eight dinoflagellate genera namely Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis and Pyrophacus were recorded Another 72 genera were belongs to diatoms Ceratium furca and a diatom Chaetoceros are the most common species found A few potentially toxic dinoflagellates species have been detected They were Dinophysis Prorocentrum and Gymnodinium The water physico-chemical parameters range from 291-322degC for temperature 26-313 PSU for salinity 73-80 for pH 21-82mgL-1 for dissolved I oxygen (DO) and 59-64 NTU for turbidity of Santubong River Result from this study showed that diatoms were dominated and dinoflagellates species composition is well distributed in all of the sampling stations
Keywords Dinoflagellates Composition Diatoms Toxic dinoflagellates Santubong River
ABSTRAK
Spesies dinojlagelat telah dikenalpasti dan dihitung melalui sampel yang telah diawet dari tiga stesen yang berlainan di sepanjang Sungai Santubong Sampel di ambil dari bulan Disember 2011 sehingga April 2012 Lapan genera dinojlagelat iaitu Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis dan Pyrophacus telah direkod Selebihnya 72 taxa merupakan diatom Spesies Ceratium furca dari dinojlagelat mendominasi kawasan tersebut diikuti oleh spesis diatom Chaetoceros Melalui kajian ini juga beberapa spesies dinojlagelat yang berpotensi untuk mengandungi toksin telah dikenalpasti Conlohnya Dinophysis Prorocentrum dan Gymnodinium Bacaan pengukur Jizikal-kimia bagi suhu ialah 291shy322 t 26-313 PSU bagi kemasinan 73-80 bagi pH 21-82mgD i bagi kandungan oksigen terlarut dan 59shy64 NTU bagi kekeruhan juga direkod di Sungai Santubong Hasil kajian mendapati diatom telah mendominasi kesemua stesen dan komposisi spesies dinojlagelat mempunyai taburan yang serata di semua stesen pensampelan
Kala kunci Komposisi Dinojlagelat Diatom Dinojlagelat bertoksik Sungai Santubong
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
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either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
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20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
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diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
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23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
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iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
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32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
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33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
i Pusat Khidmat Maklumat Akademik VNlVERSm MALAYSIA SARAWAK
TEMPORAL VARIATION IN DINOFLAGELLATE SPECIES COMPOSITION IN FISH CAGES FARM AREA ALONG SANTUBONG RIVER
PKHIDMAT MAKLUMAT AKADEMIK
111111111 fonr IIII 11111 1000235641
NURUL NADIA BINTI RAJAMI
Report Submitted in partial fulfilment for the degree of Bachelor of Science with Honours (Aquatic Resource Science and Management)
t Faculty of Resource Science and Technology
University Malaysia Sarawak 2012
I
I
bull
DECLARATION
I hereby declare that the work in this project is my own except for qoutations and summaries
which have been duly acknowledged No portion of the work referred to in this dissertation
has been summitted in support of an application for another degree qualification of this or any
other university or institution of higher learning
~
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Programme
Faculty of Resource Science and Technology
University Malaysia Sarawak
bull ~ f i
ACKNOWLEDGEMENT
Alhamdulillah Thanks to ALLAH SWT for His blessings that I finally completed my final
year project This final year project report was prepared for the partial fulfilment for the
degree of Bachelor of Science in Aquatic Resource Science and Management
I would like to express my deepest thanks to many individuals who assisted me in the
completion of this project especially to my supervisor Assoc Prof Dr Othman Bojo for his
advice motivation guidance encouragement and critics My appreciation also goes to En
Zaidi Ibrahim and other lab assistants who had helped me a lot during my sampling trips I
also want to thanks the lecturers and staffs of Faculty of Resource Science and Technology
for their cooperation during the completion of the final year project They that had given me
valuable information suggestions and guidance in the compilation and preparation this final
year project report To all my course mates thanks for the cooperation helpful suggestion and
full of support for the report completion from the beginning till the end
Finally to my beloved parents Mr Rajami bin Bujang and Mrs Sarimah binti Fadzil for being
patient thoughtful and supportive Last but not least to any individual that involved directly
and indirectly in the making of this final year project report
Thank you all
I
bull r Pusat Khidmat MakJumat Akademik UNlVERSm MALAYSIA SARAWAK
Table of Contents
ACKNOWLEDGEMENT I
T ABLE OF CONTENTS II
LIST OF ABBREVIATIONS III
LIST OF TABLES IV
LIST OF FIGURES V
ABSTRACT VI
10 INTRODUCTION
20 LITERATURE REVIEW
21 Taxonomy of Dinoflagellates 3 211 Dinoflagellates morphology 3
22 Distribution of Dinoflagellates 5 23 Ecology and Growth of Dinoflagellates 6 24 Harmful Algal Blooms (HABs) 6
241 Impacts of HABs 8
30 MATERIALS AND METHODS
31 Study Areas 9 32 Phytoplankton Sampling and Water Quality Collection 11 33 Phytoplankton identification and enumeration 12 34 Statistical Data Analysis 12
40 RESULTS
41 Qualitative Phytoplankton Composition 13 42 Dinoflagellate Assemblages 14
421 Dinoflagellates Taxonomic description 24 )43 Diatom Assemblages 30
431 Diatom Taxonomic Description 31 44 Hydro-physical Characteristics
441 pH 32 442 Temperature 33 443 Salinity 34 444 Dissolved Oxygen (DO) 35 445 Turbidity 36
50 DISCUSSION 37
60 CONCLUSIONS 43
REFERENCES 45
APPENDIX 49
I i
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LIST OF ABBREVIATIONS
~m Micro Metre
ANOVA Analysis Of Variance
APHA American Public Health Association
CFP Ciguatera Fish Poisoning
Chl-a Chlorophyll A
DNA Deoxyribonucleic Acid
DO Dissolved Oxygen
DSP Diarrhetic Shellfish Poisoning
DTXs Dinophysis Toxins
GPS Global Positioning System
HABs Harmful Algal Blooms
L Litre
Mgll Milligram Per Litre
NSP Neurotoxic Shellfish Poisoning
NTU Nephelometric Turbidity Units
PbTxs Brevetoxins
PSP Paralytic Shellfish Poisoning
PSU Practical Salinity Units
PTXs Pectenotoxins
SPSS Statistical Package For Social Science
STXs Saxitoxin
YTXs Yessotoxins
L--_ J--__ _____ _
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
List Of Tables
Sampling Sites (Station) Along The Santubong River
Taxonomic List And Occurrence Of Dinoflagellates In Santubong River
Other Phytoplankton Identified In Samples Along Santubong River
ANOVA Table For Physico-Chemical Parameters
Multiple Comparison For Physic-Chemical Parameters
The Depth Of Water Column And Its Transparency
Pages
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List Of Figures
Figure I Basic Anatomy of Thecate Dinokont Dinoflagellates
Figure 2 Maps of Santubong River
Figure 3 Percentage Distribution of Phytoplanktons
Figure 4 Number of Dinoflagellates Species Identified in Monthly Sampling
Figure 5 Number oflndividual per ml of Dinoflagellates along Santubong River
Figure 6a Representative Dinoflagellates
Figure 6b Representative Dinoflagellates
Figure 7a Representative Centric Diatoms
Figure 7b Representative Pennate Diatoms
Figure 8 Percentage Abundance of Diatoms along Santubong River
Figure 9 The Mean of Ph Value in Three Sampling Stations from December 2011 to April 2012
Figure 10 The Mean Value of Temperature in Three Sampling Stations from December 2011 to April 2012
Figure 11 The Mean Value of Salinity in Three Sampling Stations from December 2011 to April 2012
Figure 12 The Mean Value of Dissolved Oxygen CDO) in Three Sampling Stations from December 2011 to April 2012
Figure 13 The Mean Value of Turbidity in Three Sampling Stations from December 2011 to April 2012
Pages
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Temporal Variation of Dinoflagellates Species Composition at Fish Cages Area along Santubong River
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Faculty of Resources Science and Technology
University Malaysia Sarawak
ABSTRACT
Dinoflagellates were identified and counted from preserved net haul samples collected from three different stations along Santubong River The samples were collected monthly from December 20 II to Apri l 20 12 Eight dinoflagellate genera namely Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis and Pyrophacus were recorded Another 72 genera were belongs to diatoms Ceratium furca and a diatom Chaetoceros are the most common species found A few potentially toxic dinoflagellates species have been detected They were Dinophysis Prorocentrum and Gymnodinium The water physico-chemical parameters range from 291-322degC for temperature 26-313 PSU for salinity 73-80 for pH 21-82mgL-1 for dissolved I oxygen (DO) and 59-64 NTU for turbidity of Santubong River Result from this study showed that diatoms were dominated and dinoflagellates species composition is well distributed in all of the sampling stations
Keywords Dinoflagellates Composition Diatoms Toxic dinoflagellates Santubong River
ABSTRAK
Spesies dinojlagelat telah dikenalpasti dan dihitung melalui sampel yang telah diawet dari tiga stesen yang berlainan di sepanjang Sungai Santubong Sampel di ambil dari bulan Disember 2011 sehingga April 2012 Lapan genera dinojlagelat iaitu Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis dan Pyrophacus telah direkod Selebihnya 72 taxa merupakan diatom Spesies Ceratium furca dari dinojlagelat mendominasi kawasan tersebut diikuti oleh spesis diatom Chaetoceros Melalui kajian ini juga beberapa spesies dinojlagelat yang berpotensi untuk mengandungi toksin telah dikenalpasti Conlohnya Dinophysis Prorocentrum dan Gymnodinium Bacaan pengukur Jizikal-kimia bagi suhu ialah 291shy322 t 26-313 PSU bagi kemasinan 73-80 bagi pH 21-82mgD i bagi kandungan oksigen terlarut dan 59shy64 NTU bagi kekeruhan juga direkod di Sungai Santubong Hasil kajian mendapati diatom telah mendominasi kesemua stesen dan komposisi spesies dinojlagelat mempunyai taburan yang serata di semua stesen pensampelan
Kala kunci Komposisi Dinojlagelat Diatom Dinojlagelat bertoksik Sungai Santubong
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
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either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
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20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
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diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
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23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
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iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
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32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
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33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
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DECLARATION
I hereby declare that the work in this project is my own except for qoutations and summaries
which have been duly acknowledged No portion of the work referred to in this dissertation
has been summitted in support of an application for another degree qualification of this or any
other university or institution of higher learning
~
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Programme
Faculty of Resource Science and Technology
University Malaysia Sarawak
bull ~ f i
ACKNOWLEDGEMENT
Alhamdulillah Thanks to ALLAH SWT for His blessings that I finally completed my final
year project This final year project report was prepared for the partial fulfilment for the
degree of Bachelor of Science in Aquatic Resource Science and Management
I would like to express my deepest thanks to many individuals who assisted me in the
completion of this project especially to my supervisor Assoc Prof Dr Othman Bojo for his
advice motivation guidance encouragement and critics My appreciation also goes to En
Zaidi Ibrahim and other lab assistants who had helped me a lot during my sampling trips I
also want to thanks the lecturers and staffs of Faculty of Resource Science and Technology
for their cooperation during the completion of the final year project They that had given me
valuable information suggestions and guidance in the compilation and preparation this final
year project report To all my course mates thanks for the cooperation helpful suggestion and
full of support for the report completion from the beginning till the end
Finally to my beloved parents Mr Rajami bin Bujang and Mrs Sarimah binti Fadzil for being
patient thoughtful and supportive Last but not least to any individual that involved directly
and indirectly in the making of this final year project report
Thank you all
I
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Table of Contents
ACKNOWLEDGEMENT I
T ABLE OF CONTENTS II
LIST OF ABBREVIATIONS III
LIST OF TABLES IV
LIST OF FIGURES V
ABSTRACT VI
10 INTRODUCTION
20 LITERATURE REVIEW
21 Taxonomy of Dinoflagellates 3 211 Dinoflagellates morphology 3
22 Distribution of Dinoflagellates 5 23 Ecology and Growth of Dinoflagellates 6 24 Harmful Algal Blooms (HABs) 6
241 Impacts of HABs 8
30 MATERIALS AND METHODS
31 Study Areas 9 32 Phytoplankton Sampling and Water Quality Collection 11 33 Phytoplankton identification and enumeration 12 34 Statistical Data Analysis 12
40 RESULTS
41 Qualitative Phytoplankton Composition 13 42 Dinoflagellate Assemblages 14
421 Dinoflagellates Taxonomic description 24 )43 Diatom Assemblages 30
431 Diatom Taxonomic Description 31 44 Hydro-physical Characteristics
441 pH 32 442 Temperature 33 443 Salinity 34 444 Dissolved Oxygen (DO) 35 445 Turbidity 36
50 DISCUSSION 37
60 CONCLUSIONS 43
REFERENCES 45
APPENDIX 49
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i
LIST OF ABBREVIATIONS
~m Micro Metre
ANOVA Analysis Of Variance
APHA American Public Health Association
CFP Ciguatera Fish Poisoning
Chl-a Chlorophyll A
DNA Deoxyribonucleic Acid
DO Dissolved Oxygen
DSP Diarrhetic Shellfish Poisoning
DTXs Dinophysis Toxins
GPS Global Positioning System
HABs Harmful Algal Blooms
L Litre
Mgll Milligram Per Litre
NSP Neurotoxic Shellfish Poisoning
NTU Nephelometric Turbidity Units
PbTxs Brevetoxins
PSP Paralytic Shellfish Poisoning
PSU Practical Salinity Units
PTXs Pectenotoxins
SPSS Statistical Package For Social Science
STXs Saxitoxin
YTXs Yessotoxins
L--_ J--__ _____ _
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
List Of Tables
Sampling Sites (Station) Along The Santubong River
Taxonomic List And Occurrence Of Dinoflagellates In Santubong River
Other Phytoplankton Identified In Samples Along Santubong River
ANOVA Table For Physico-Chemical Parameters
Multiple Comparison For Physic-Chemical Parameters
The Depth Of Water Column And Its Transparency
Pages
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List Of Figures
Figure I Basic Anatomy of Thecate Dinokont Dinoflagellates
Figure 2 Maps of Santubong River
Figure 3 Percentage Distribution of Phytoplanktons
Figure 4 Number of Dinoflagellates Species Identified in Monthly Sampling
Figure 5 Number oflndividual per ml of Dinoflagellates along Santubong River
Figure 6a Representative Dinoflagellates
Figure 6b Representative Dinoflagellates
Figure 7a Representative Centric Diatoms
Figure 7b Representative Pennate Diatoms
Figure 8 Percentage Abundance of Diatoms along Santubong River
Figure 9 The Mean of Ph Value in Three Sampling Stations from December 2011 to April 2012
Figure 10 The Mean Value of Temperature in Three Sampling Stations from December 2011 to April 2012
Figure 11 The Mean Value of Salinity in Three Sampling Stations from December 2011 to April 2012
Figure 12 The Mean Value of Dissolved Oxygen CDO) in Three Sampling Stations from December 2011 to April 2012
Figure 13 The Mean Value of Turbidity in Three Sampling Stations from December 2011 to April 2012
Pages
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Temporal Variation of Dinoflagellates Species Composition at Fish Cages Area along Santubong River
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Faculty of Resources Science and Technology
University Malaysia Sarawak
ABSTRACT
Dinoflagellates were identified and counted from preserved net haul samples collected from three different stations along Santubong River The samples were collected monthly from December 20 II to Apri l 20 12 Eight dinoflagellate genera namely Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis and Pyrophacus were recorded Another 72 genera were belongs to diatoms Ceratium furca and a diatom Chaetoceros are the most common species found A few potentially toxic dinoflagellates species have been detected They were Dinophysis Prorocentrum and Gymnodinium The water physico-chemical parameters range from 291-322degC for temperature 26-313 PSU for salinity 73-80 for pH 21-82mgL-1 for dissolved I oxygen (DO) and 59-64 NTU for turbidity of Santubong River Result from this study showed that diatoms were dominated and dinoflagellates species composition is well distributed in all of the sampling stations
Keywords Dinoflagellates Composition Diatoms Toxic dinoflagellates Santubong River
ABSTRAK
Spesies dinojlagelat telah dikenalpasti dan dihitung melalui sampel yang telah diawet dari tiga stesen yang berlainan di sepanjang Sungai Santubong Sampel di ambil dari bulan Disember 2011 sehingga April 2012 Lapan genera dinojlagelat iaitu Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis dan Pyrophacus telah direkod Selebihnya 72 taxa merupakan diatom Spesies Ceratium furca dari dinojlagelat mendominasi kawasan tersebut diikuti oleh spesis diatom Chaetoceros Melalui kajian ini juga beberapa spesies dinojlagelat yang berpotensi untuk mengandungi toksin telah dikenalpasti Conlohnya Dinophysis Prorocentrum dan Gymnodinium Bacaan pengukur Jizikal-kimia bagi suhu ialah 291shy322 t 26-313 PSU bagi kemasinan 73-80 bagi pH 21-82mgD i bagi kandungan oksigen terlarut dan 59shy64 NTU bagi kekeruhan juga direkod di Sungai Santubong Hasil kajian mendapati diatom telah mendominasi kesemua stesen dan komposisi spesies dinojlagelat mempunyai taburan yang serata di semua stesen pensampelan
Kala kunci Komposisi Dinojlagelat Diatom Dinojlagelat bertoksik Sungai Santubong
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
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either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
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20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
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diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
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23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
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iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
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32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
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33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
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ACKNOWLEDGEMENT
Alhamdulillah Thanks to ALLAH SWT for His blessings that I finally completed my final
year project This final year project report was prepared for the partial fulfilment for the
degree of Bachelor of Science in Aquatic Resource Science and Management
I would like to express my deepest thanks to many individuals who assisted me in the
completion of this project especially to my supervisor Assoc Prof Dr Othman Bojo for his
advice motivation guidance encouragement and critics My appreciation also goes to En
Zaidi Ibrahim and other lab assistants who had helped me a lot during my sampling trips I
also want to thanks the lecturers and staffs of Faculty of Resource Science and Technology
for their cooperation during the completion of the final year project They that had given me
valuable information suggestions and guidance in the compilation and preparation this final
year project report To all my course mates thanks for the cooperation helpful suggestion and
full of support for the report completion from the beginning till the end
Finally to my beloved parents Mr Rajami bin Bujang and Mrs Sarimah binti Fadzil for being
patient thoughtful and supportive Last but not least to any individual that involved directly
and indirectly in the making of this final year project report
Thank you all
I
bull r Pusat Khidmat MakJumat Akademik UNlVERSm MALAYSIA SARAWAK
Table of Contents
ACKNOWLEDGEMENT I
T ABLE OF CONTENTS II
LIST OF ABBREVIATIONS III
LIST OF TABLES IV
LIST OF FIGURES V
ABSTRACT VI
10 INTRODUCTION
20 LITERATURE REVIEW
21 Taxonomy of Dinoflagellates 3 211 Dinoflagellates morphology 3
22 Distribution of Dinoflagellates 5 23 Ecology and Growth of Dinoflagellates 6 24 Harmful Algal Blooms (HABs) 6
241 Impacts of HABs 8
30 MATERIALS AND METHODS
31 Study Areas 9 32 Phytoplankton Sampling and Water Quality Collection 11 33 Phytoplankton identification and enumeration 12 34 Statistical Data Analysis 12
40 RESULTS
41 Qualitative Phytoplankton Composition 13 42 Dinoflagellate Assemblages 14
421 Dinoflagellates Taxonomic description 24 )43 Diatom Assemblages 30
431 Diatom Taxonomic Description 31 44 Hydro-physical Characteristics
441 pH 32 442 Temperature 33 443 Salinity 34 444 Dissolved Oxygen (DO) 35 445 Turbidity 36
50 DISCUSSION 37
60 CONCLUSIONS 43
REFERENCES 45
APPENDIX 49
I i
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i
LIST OF ABBREVIATIONS
~m Micro Metre
ANOVA Analysis Of Variance
APHA American Public Health Association
CFP Ciguatera Fish Poisoning
Chl-a Chlorophyll A
DNA Deoxyribonucleic Acid
DO Dissolved Oxygen
DSP Diarrhetic Shellfish Poisoning
DTXs Dinophysis Toxins
GPS Global Positioning System
HABs Harmful Algal Blooms
L Litre
Mgll Milligram Per Litre
NSP Neurotoxic Shellfish Poisoning
NTU Nephelometric Turbidity Units
PbTxs Brevetoxins
PSP Paralytic Shellfish Poisoning
PSU Practical Salinity Units
PTXs Pectenotoxins
SPSS Statistical Package For Social Science
STXs Saxitoxin
YTXs Yessotoxins
L--_ J--__ _____ _
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
List Of Tables
Sampling Sites (Station) Along The Santubong River
Taxonomic List And Occurrence Of Dinoflagellates In Santubong River
Other Phytoplankton Identified In Samples Along Santubong River
ANOVA Table For Physico-Chemical Parameters
Multiple Comparison For Physic-Chemical Parameters
The Depth Of Water Column And Its Transparency
Pages
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20
21
49
50
52
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List Of Figures
Figure I Basic Anatomy of Thecate Dinokont Dinoflagellates
Figure 2 Maps of Santubong River
Figure 3 Percentage Distribution of Phytoplanktons
Figure 4 Number of Dinoflagellates Species Identified in Monthly Sampling
Figure 5 Number oflndividual per ml of Dinoflagellates along Santubong River
Figure 6a Representative Dinoflagellates
Figure 6b Representative Dinoflagellates
Figure 7a Representative Centric Diatoms
Figure 7b Representative Pennate Diatoms
Figure 8 Percentage Abundance of Diatoms along Santubong River
Figure 9 The Mean of Ph Value in Three Sampling Stations from December 2011 to April 2012
Figure 10 The Mean Value of Temperature in Three Sampling Stations from December 2011 to April 2012
Figure 11 The Mean Value of Salinity in Three Sampling Stations from December 2011 to April 2012
Figure 12 The Mean Value of Dissolved Oxygen CDO) in Three Sampling Stations from December 2011 to April 2012
Figure 13 The Mean Value of Turbidity in Three Sampling Stations from December 2011 to April 2012
Pages
4
10
13
15
15
16
17 ~
18
19
30
32
33
34
35
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Temporal Variation of Dinoflagellates Species Composition at Fish Cages Area along Santubong River
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Faculty of Resources Science and Technology
University Malaysia Sarawak
ABSTRACT
Dinoflagellates were identified and counted from preserved net haul samples collected from three different stations along Santubong River The samples were collected monthly from December 20 II to Apri l 20 12 Eight dinoflagellate genera namely Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis and Pyrophacus were recorded Another 72 genera were belongs to diatoms Ceratium furca and a diatom Chaetoceros are the most common species found A few potentially toxic dinoflagellates species have been detected They were Dinophysis Prorocentrum and Gymnodinium The water physico-chemical parameters range from 291-322degC for temperature 26-313 PSU for salinity 73-80 for pH 21-82mgL-1 for dissolved I oxygen (DO) and 59-64 NTU for turbidity of Santubong River Result from this study showed that diatoms were dominated and dinoflagellates species composition is well distributed in all of the sampling stations
Keywords Dinoflagellates Composition Diatoms Toxic dinoflagellates Santubong River
ABSTRAK
Spesies dinojlagelat telah dikenalpasti dan dihitung melalui sampel yang telah diawet dari tiga stesen yang berlainan di sepanjang Sungai Santubong Sampel di ambil dari bulan Disember 2011 sehingga April 2012 Lapan genera dinojlagelat iaitu Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis dan Pyrophacus telah direkod Selebihnya 72 taxa merupakan diatom Spesies Ceratium furca dari dinojlagelat mendominasi kawasan tersebut diikuti oleh spesis diatom Chaetoceros Melalui kajian ini juga beberapa spesies dinojlagelat yang berpotensi untuk mengandungi toksin telah dikenalpasti Conlohnya Dinophysis Prorocentrum dan Gymnodinium Bacaan pengukur Jizikal-kimia bagi suhu ialah 291shy322 t 26-313 PSU bagi kemasinan 73-80 bagi pH 21-82mgD i bagi kandungan oksigen terlarut dan 59shy64 NTU bagi kekeruhan juga direkod di Sungai Santubong Hasil kajian mendapati diatom telah mendominasi kesemua stesen dan komposisi spesies dinojlagelat mempunyai taburan yang serata di semua stesen pensampelan
Kala kunci Komposisi Dinojlagelat Diatom Dinojlagelat bertoksik Sungai Santubong
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
It bull
either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
I
20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
l
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
I
bull r Pusat Khidmat MakJumat Akademik UNlVERSm MALAYSIA SARAWAK
Table of Contents
ACKNOWLEDGEMENT I
T ABLE OF CONTENTS II
LIST OF ABBREVIATIONS III
LIST OF TABLES IV
LIST OF FIGURES V
ABSTRACT VI
10 INTRODUCTION
20 LITERATURE REVIEW
21 Taxonomy of Dinoflagellates 3 211 Dinoflagellates morphology 3
22 Distribution of Dinoflagellates 5 23 Ecology and Growth of Dinoflagellates 6 24 Harmful Algal Blooms (HABs) 6
241 Impacts of HABs 8
30 MATERIALS AND METHODS
31 Study Areas 9 32 Phytoplankton Sampling and Water Quality Collection 11 33 Phytoplankton identification and enumeration 12 34 Statistical Data Analysis 12
40 RESULTS
41 Qualitative Phytoplankton Composition 13 42 Dinoflagellate Assemblages 14
421 Dinoflagellates Taxonomic description 24 )43 Diatom Assemblages 30
431 Diatom Taxonomic Description 31 44 Hydro-physical Characteristics
441 pH 32 442 Temperature 33 443 Salinity 34 444 Dissolved Oxygen (DO) 35 445 Turbidity 36
50 DISCUSSION 37
60 CONCLUSIONS 43
REFERENCES 45
APPENDIX 49
I i
I
i
LIST OF ABBREVIATIONS
~m Micro Metre
ANOVA Analysis Of Variance
APHA American Public Health Association
CFP Ciguatera Fish Poisoning
Chl-a Chlorophyll A
DNA Deoxyribonucleic Acid
DO Dissolved Oxygen
DSP Diarrhetic Shellfish Poisoning
DTXs Dinophysis Toxins
GPS Global Positioning System
HABs Harmful Algal Blooms
L Litre
Mgll Milligram Per Litre
NSP Neurotoxic Shellfish Poisoning
NTU Nephelometric Turbidity Units
PbTxs Brevetoxins
PSP Paralytic Shellfish Poisoning
PSU Practical Salinity Units
PTXs Pectenotoxins
SPSS Statistical Package For Social Science
STXs Saxitoxin
YTXs Yessotoxins
L--_ J--__ _____ _
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
List Of Tables
Sampling Sites (Station) Along The Santubong River
Taxonomic List And Occurrence Of Dinoflagellates In Santubong River
Other Phytoplankton Identified In Samples Along Santubong River
ANOVA Table For Physico-Chemical Parameters
Multiple Comparison For Physic-Chemical Parameters
The Depth Of Water Column And Its Transparency
Pages
9
20
21
49
50
52
I
bull t f
List Of Figures
Figure I Basic Anatomy of Thecate Dinokont Dinoflagellates
Figure 2 Maps of Santubong River
Figure 3 Percentage Distribution of Phytoplanktons
Figure 4 Number of Dinoflagellates Species Identified in Monthly Sampling
Figure 5 Number oflndividual per ml of Dinoflagellates along Santubong River
Figure 6a Representative Dinoflagellates
Figure 6b Representative Dinoflagellates
Figure 7a Representative Centric Diatoms
Figure 7b Representative Pennate Diatoms
Figure 8 Percentage Abundance of Diatoms along Santubong River
Figure 9 The Mean of Ph Value in Three Sampling Stations from December 2011 to April 2012
Figure 10 The Mean Value of Temperature in Three Sampling Stations from December 2011 to April 2012
Figure 11 The Mean Value of Salinity in Three Sampling Stations from December 2011 to April 2012
Figure 12 The Mean Value of Dissolved Oxygen CDO) in Three Sampling Stations from December 2011 to April 2012
Figure 13 The Mean Value of Turbidity in Three Sampling Stations from December 2011 to April 2012
Pages
4
10
13
15
15
16
17 ~
18
19
30
32
33
34
35
36
bull bull
Temporal Variation of Dinoflagellates Species Composition at Fish Cages Area along Santubong River
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Faculty of Resources Science and Technology
University Malaysia Sarawak
ABSTRACT
Dinoflagellates were identified and counted from preserved net haul samples collected from three different stations along Santubong River The samples were collected monthly from December 20 II to Apri l 20 12 Eight dinoflagellate genera namely Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis and Pyrophacus were recorded Another 72 genera were belongs to diatoms Ceratium furca and a diatom Chaetoceros are the most common species found A few potentially toxic dinoflagellates species have been detected They were Dinophysis Prorocentrum and Gymnodinium The water physico-chemical parameters range from 291-322degC for temperature 26-313 PSU for salinity 73-80 for pH 21-82mgL-1 for dissolved I oxygen (DO) and 59-64 NTU for turbidity of Santubong River Result from this study showed that diatoms were dominated and dinoflagellates species composition is well distributed in all of the sampling stations
Keywords Dinoflagellates Composition Diatoms Toxic dinoflagellates Santubong River
ABSTRAK
Spesies dinojlagelat telah dikenalpasti dan dihitung melalui sampel yang telah diawet dari tiga stesen yang berlainan di sepanjang Sungai Santubong Sampel di ambil dari bulan Disember 2011 sehingga April 2012 Lapan genera dinojlagelat iaitu Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis dan Pyrophacus telah direkod Selebihnya 72 taxa merupakan diatom Spesies Ceratium furca dari dinojlagelat mendominasi kawasan tersebut diikuti oleh spesis diatom Chaetoceros Melalui kajian ini juga beberapa spesies dinojlagelat yang berpotensi untuk mengandungi toksin telah dikenalpasti Conlohnya Dinophysis Prorocentrum dan Gymnodinium Bacaan pengukur Jizikal-kimia bagi suhu ialah 291shy322 t 26-313 PSU bagi kemasinan 73-80 bagi pH 21-82mgD i bagi kandungan oksigen terlarut dan 59shy64 NTU bagi kekeruhan juga direkod di Sungai Santubong Hasil kajian mendapati diatom telah mendominasi kesemua stesen dan komposisi spesies dinojlagelat mempunyai taburan yang serata di semua stesen pensampelan
Kala kunci Komposisi Dinojlagelat Diatom Dinojlagelat bertoksik Sungai Santubong
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
It bull
either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
I
20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
l
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
I i
I
i
LIST OF ABBREVIATIONS
~m Micro Metre
ANOVA Analysis Of Variance
APHA American Public Health Association
CFP Ciguatera Fish Poisoning
Chl-a Chlorophyll A
DNA Deoxyribonucleic Acid
DO Dissolved Oxygen
DSP Diarrhetic Shellfish Poisoning
DTXs Dinophysis Toxins
GPS Global Positioning System
HABs Harmful Algal Blooms
L Litre
Mgll Milligram Per Litre
NSP Neurotoxic Shellfish Poisoning
NTU Nephelometric Turbidity Units
PbTxs Brevetoxins
PSP Paralytic Shellfish Poisoning
PSU Practical Salinity Units
PTXs Pectenotoxins
SPSS Statistical Package For Social Science
STXs Saxitoxin
YTXs Yessotoxins
L--_ J--__ _____ _
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
List Of Tables
Sampling Sites (Station) Along The Santubong River
Taxonomic List And Occurrence Of Dinoflagellates In Santubong River
Other Phytoplankton Identified In Samples Along Santubong River
ANOVA Table For Physico-Chemical Parameters
Multiple Comparison For Physic-Chemical Parameters
The Depth Of Water Column And Its Transparency
Pages
9
20
21
49
50
52
I
bull t f
List Of Figures
Figure I Basic Anatomy of Thecate Dinokont Dinoflagellates
Figure 2 Maps of Santubong River
Figure 3 Percentage Distribution of Phytoplanktons
Figure 4 Number of Dinoflagellates Species Identified in Monthly Sampling
Figure 5 Number oflndividual per ml of Dinoflagellates along Santubong River
Figure 6a Representative Dinoflagellates
Figure 6b Representative Dinoflagellates
Figure 7a Representative Centric Diatoms
Figure 7b Representative Pennate Diatoms
Figure 8 Percentage Abundance of Diatoms along Santubong River
Figure 9 The Mean of Ph Value in Three Sampling Stations from December 2011 to April 2012
Figure 10 The Mean Value of Temperature in Three Sampling Stations from December 2011 to April 2012
Figure 11 The Mean Value of Salinity in Three Sampling Stations from December 2011 to April 2012
Figure 12 The Mean Value of Dissolved Oxygen CDO) in Three Sampling Stations from December 2011 to April 2012
Figure 13 The Mean Value of Turbidity in Three Sampling Stations from December 2011 to April 2012
Pages
4
10
13
15
15
16
17 ~
18
19
30
32
33
34
35
36
bull bull
Temporal Variation of Dinoflagellates Species Composition at Fish Cages Area along Santubong River
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Faculty of Resources Science and Technology
University Malaysia Sarawak
ABSTRACT
Dinoflagellates were identified and counted from preserved net haul samples collected from three different stations along Santubong River The samples were collected monthly from December 20 II to Apri l 20 12 Eight dinoflagellate genera namely Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis and Pyrophacus were recorded Another 72 genera were belongs to diatoms Ceratium furca and a diatom Chaetoceros are the most common species found A few potentially toxic dinoflagellates species have been detected They were Dinophysis Prorocentrum and Gymnodinium The water physico-chemical parameters range from 291-322degC for temperature 26-313 PSU for salinity 73-80 for pH 21-82mgL-1 for dissolved I oxygen (DO) and 59-64 NTU for turbidity of Santubong River Result from this study showed that diatoms were dominated and dinoflagellates species composition is well distributed in all of the sampling stations
Keywords Dinoflagellates Composition Diatoms Toxic dinoflagellates Santubong River
ABSTRAK
Spesies dinojlagelat telah dikenalpasti dan dihitung melalui sampel yang telah diawet dari tiga stesen yang berlainan di sepanjang Sungai Santubong Sampel di ambil dari bulan Disember 2011 sehingga April 2012 Lapan genera dinojlagelat iaitu Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis dan Pyrophacus telah direkod Selebihnya 72 taxa merupakan diatom Spesies Ceratium furca dari dinojlagelat mendominasi kawasan tersebut diikuti oleh spesis diatom Chaetoceros Melalui kajian ini juga beberapa spesies dinojlagelat yang berpotensi untuk mengandungi toksin telah dikenalpasti Conlohnya Dinophysis Prorocentrum dan Gymnodinium Bacaan pengukur Jizikal-kimia bagi suhu ialah 291shy322 t 26-313 PSU bagi kemasinan 73-80 bagi pH 21-82mgD i bagi kandungan oksigen terlarut dan 59shy64 NTU bagi kekeruhan juga direkod di Sungai Santubong Hasil kajian mendapati diatom telah mendominasi kesemua stesen dan komposisi spesies dinojlagelat mempunyai taburan yang serata di semua stesen pensampelan
Kala kunci Komposisi Dinojlagelat Diatom Dinojlagelat bertoksik Sungai Santubong
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
It bull
either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
I
20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
l
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
List Of Tables
Sampling Sites (Station) Along The Santubong River
Taxonomic List And Occurrence Of Dinoflagellates In Santubong River
Other Phytoplankton Identified In Samples Along Santubong River
ANOVA Table For Physico-Chemical Parameters
Multiple Comparison For Physic-Chemical Parameters
The Depth Of Water Column And Its Transparency
Pages
9
20
21
49
50
52
I
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List Of Figures
Figure I Basic Anatomy of Thecate Dinokont Dinoflagellates
Figure 2 Maps of Santubong River
Figure 3 Percentage Distribution of Phytoplanktons
Figure 4 Number of Dinoflagellates Species Identified in Monthly Sampling
Figure 5 Number oflndividual per ml of Dinoflagellates along Santubong River
Figure 6a Representative Dinoflagellates
Figure 6b Representative Dinoflagellates
Figure 7a Representative Centric Diatoms
Figure 7b Representative Pennate Diatoms
Figure 8 Percentage Abundance of Diatoms along Santubong River
Figure 9 The Mean of Ph Value in Three Sampling Stations from December 2011 to April 2012
Figure 10 The Mean Value of Temperature in Three Sampling Stations from December 2011 to April 2012
Figure 11 The Mean Value of Salinity in Three Sampling Stations from December 2011 to April 2012
Figure 12 The Mean Value of Dissolved Oxygen CDO) in Three Sampling Stations from December 2011 to April 2012
Figure 13 The Mean Value of Turbidity in Three Sampling Stations from December 2011 to April 2012
Pages
4
10
13
15
15
16
17 ~
18
19
30
32
33
34
35
36
bull bull
Temporal Variation of Dinoflagellates Species Composition at Fish Cages Area along Santubong River
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Faculty of Resources Science and Technology
University Malaysia Sarawak
ABSTRACT
Dinoflagellates were identified and counted from preserved net haul samples collected from three different stations along Santubong River The samples were collected monthly from December 20 II to Apri l 20 12 Eight dinoflagellate genera namely Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis and Pyrophacus were recorded Another 72 genera were belongs to diatoms Ceratium furca and a diatom Chaetoceros are the most common species found A few potentially toxic dinoflagellates species have been detected They were Dinophysis Prorocentrum and Gymnodinium The water physico-chemical parameters range from 291-322degC for temperature 26-313 PSU for salinity 73-80 for pH 21-82mgL-1 for dissolved I oxygen (DO) and 59-64 NTU for turbidity of Santubong River Result from this study showed that diatoms were dominated and dinoflagellates species composition is well distributed in all of the sampling stations
Keywords Dinoflagellates Composition Diatoms Toxic dinoflagellates Santubong River
ABSTRAK
Spesies dinojlagelat telah dikenalpasti dan dihitung melalui sampel yang telah diawet dari tiga stesen yang berlainan di sepanjang Sungai Santubong Sampel di ambil dari bulan Disember 2011 sehingga April 2012 Lapan genera dinojlagelat iaitu Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis dan Pyrophacus telah direkod Selebihnya 72 taxa merupakan diatom Spesies Ceratium furca dari dinojlagelat mendominasi kawasan tersebut diikuti oleh spesis diatom Chaetoceros Melalui kajian ini juga beberapa spesies dinojlagelat yang berpotensi untuk mengandungi toksin telah dikenalpasti Conlohnya Dinophysis Prorocentrum dan Gymnodinium Bacaan pengukur Jizikal-kimia bagi suhu ialah 291shy322 t 26-313 PSU bagi kemasinan 73-80 bagi pH 21-82mgD i bagi kandungan oksigen terlarut dan 59shy64 NTU bagi kekeruhan juga direkod di Sungai Santubong Hasil kajian mendapati diatom telah mendominasi kesemua stesen dan komposisi spesies dinojlagelat mempunyai taburan yang serata di semua stesen pensampelan
Kala kunci Komposisi Dinojlagelat Diatom Dinojlagelat bertoksik Sungai Santubong
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
It bull
either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
I
20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
l
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
I
bull t f
List Of Figures
Figure I Basic Anatomy of Thecate Dinokont Dinoflagellates
Figure 2 Maps of Santubong River
Figure 3 Percentage Distribution of Phytoplanktons
Figure 4 Number of Dinoflagellates Species Identified in Monthly Sampling
Figure 5 Number oflndividual per ml of Dinoflagellates along Santubong River
Figure 6a Representative Dinoflagellates
Figure 6b Representative Dinoflagellates
Figure 7a Representative Centric Diatoms
Figure 7b Representative Pennate Diatoms
Figure 8 Percentage Abundance of Diatoms along Santubong River
Figure 9 The Mean of Ph Value in Three Sampling Stations from December 2011 to April 2012
Figure 10 The Mean Value of Temperature in Three Sampling Stations from December 2011 to April 2012
Figure 11 The Mean Value of Salinity in Three Sampling Stations from December 2011 to April 2012
Figure 12 The Mean Value of Dissolved Oxygen CDO) in Three Sampling Stations from December 2011 to April 2012
Figure 13 The Mean Value of Turbidity in Three Sampling Stations from December 2011 to April 2012
Pages
4
10
13
15
15
16
17 ~
18
19
30
32
33
34
35
36
bull bull
Temporal Variation of Dinoflagellates Species Composition at Fish Cages Area along Santubong River
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Faculty of Resources Science and Technology
University Malaysia Sarawak
ABSTRACT
Dinoflagellates were identified and counted from preserved net haul samples collected from three different stations along Santubong River The samples were collected monthly from December 20 II to Apri l 20 12 Eight dinoflagellate genera namely Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis and Pyrophacus were recorded Another 72 genera were belongs to diatoms Ceratium furca and a diatom Chaetoceros are the most common species found A few potentially toxic dinoflagellates species have been detected They were Dinophysis Prorocentrum and Gymnodinium The water physico-chemical parameters range from 291-322degC for temperature 26-313 PSU for salinity 73-80 for pH 21-82mgL-1 for dissolved I oxygen (DO) and 59-64 NTU for turbidity of Santubong River Result from this study showed that diatoms were dominated and dinoflagellates species composition is well distributed in all of the sampling stations
Keywords Dinoflagellates Composition Diatoms Toxic dinoflagellates Santubong River
ABSTRAK
Spesies dinojlagelat telah dikenalpasti dan dihitung melalui sampel yang telah diawet dari tiga stesen yang berlainan di sepanjang Sungai Santubong Sampel di ambil dari bulan Disember 2011 sehingga April 2012 Lapan genera dinojlagelat iaitu Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis dan Pyrophacus telah direkod Selebihnya 72 taxa merupakan diatom Spesies Ceratium furca dari dinojlagelat mendominasi kawasan tersebut diikuti oleh spesis diatom Chaetoceros Melalui kajian ini juga beberapa spesies dinojlagelat yang berpotensi untuk mengandungi toksin telah dikenalpasti Conlohnya Dinophysis Prorocentrum dan Gymnodinium Bacaan pengukur Jizikal-kimia bagi suhu ialah 291shy322 t 26-313 PSU bagi kemasinan 73-80 bagi pH 21-82mgD i bagi kandungan oksigen terlarut dan 59shy64 NTU bagi kekeruhan juga direkod di Sungai Santubong Hasil kajian mendapati diatom telah mendominasi kesemua stesen dan komposisi spesies dinojlagelat mempunyai taburan yang serata di semua stesen pensampelan
Kala kunci Komposisi Dinojlagelat Diatom Dinojlagelat bertoksik Sungai Santubong
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
It bull
either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
I
20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
l
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
bull bull
Temporal Variation of Dinoflagellates Species Composition at Fish Cages Area along Santubong River
Nurul Nadia binti Rajami
Aquatic Resources Science and Management Faculty of Resources Science and Technology
University Malaysia Sarawak
ABSTRACT
Dinoflagellates were identified and counted from preserved net haul samples collected from three different stations along Santubong River The samples were collected monthly from December 20 II to Apri l 20 12 Eight dinoflagellate genera namely Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis and Pyrophacus were recorded Another 72 genera were belongs to diatoms Ceratium furca and a diatom Chaetoceros are the most common species found A few potentially toxic dinoflagellates species have been detected They were Dinophysis Prorocentrum and Gymnodinium The water physico-chemical parameters range from 291-322degC for temperature 26-313 PSU for salinity 73-80 for pH 21-82mgL-1 for dissolved I oxygen (DO) and 59-64 NTU for turbidity of Santubong River Result from this study showed that diatoms were dominated and dinoflagellates species composition is well distributed in all of the sampling stations
Keywords Dinoflagellates Composition Diatoms Toxic dinoflagellates Santubong River
ABSTRAK
Spesies dinojlagelat telah dikenalpasti dan dihitung melalui sampel yang telah diawet dari tiga stesen yang berlainan di sepanjang Sungai Santubong Sampel di ambil dari bulan Disember 2011 sehingga April 2012 Lapan genera dinojlagelat iaitu Ceratium Dinophysis Gymnodinium Gonyaulax Prorocentrum Protoperidinium Pyrocystis dan Pyrophacus telah direkod Selebihnya 72 taxa merupakan diatom Spesies Ceratium furca dari dinojlagelat mendominasi kawasan tersebut diikuti oleh spesis diatom Chaetoceros Melalui kajian ini juga beberapa spesies dinojlagelat yang berpotensi untuk mengandungi toksin telah dikenalpasti Conlohnya Dinophysis Prorocentrum dan Gymnodinium Bacaan pengukur Jizikal-kimia bagi suhu ialah 291shy322 t 26-313 PSU bagi kemasinan 73-80 bagi pH 21-82mgD i bagi kandungan oksigen terlarut dan 59shy64 NTU bagi kekeruhan juga direkod di Sungai Santubong Hasil kajian mendapati diatom telah mendominasi kesemua stesen dan komposisi spesies dinojlagelat mempunyai taburan yang serata di semua stesen pensampelan
Kala kunci Komposisi Dinojlagelat Diatom Dinojlagelat bertoksik Sungai Santubong
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
It bull
either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
I
20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
l
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
10 INTRODUCTION
Dinoflagellates are one of the most important members of the phytoplankton in
marine and freshwater ecosystems which represent a major component in food webs
(Saldiarriaga amp Hoppenrath 2010) Their cell sizes were range from 2 Jlm to 20 Jlm More
than 2000 existing species have been described only half of which are photosynthetic
They include autotrophs mixotrophs and grazers (Taylor et ai 2007) Most of the
photosynthetic zooxanthellae of invertebrate hosts are mutualistic dinoflagellate
symbionts plus all those crucial to reef-building corals (Taylor et ai 2007) Some of them
are also the important sources of bioluminescence in the ocean which can light up at night
(Hallegraeff 1995) Beside diatoms over half of the present-day dinoflagellates are
photosynthetic and are at the base of marine food web (Spector 1984) Furthermore
dinoflagellate gives food to filter-feeding bivalve shellfish such as oysters mussels
scallops and clams (Faust 2002)
Dinoflagellates can be identified by the presence of dimorphic flagella one
flagellum oriented around the cell and the other directed posterior which allow them to
swim freely in water with a forward spiralling motion (Faust 2002) Another characteristic
of the dinoflagellates is the wall configuration and arrangement that was based on the
presence (armoured) or absence (unarmoured) of a rigid outer cell covering or also known
as theca The dinoflagellates have unique characteristics It contains a distinctive nucleus
named the dinokaryon The DNA is not systematized around nucleosomal histones as a
substitute it forming fibrillar chromosomes that are always condensed and that divide via a
closed mitosis with an external spindle (Taylor et ai 2007)
When dinoflagellate proliferates to higher than normal concentrations the water
may discolour and appears red This condition is called red tides The blooms may be
1
It bull
either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
I
20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
l
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
It bull
either toxic or noxious which they often accumulate in shellfish or fish and when these are
eaten by humans they cause diseases like paralytic shellfish poisoning (PSP) neurotoxic
shellfish poisoning (NSP) diarrhetic shellfish poisoning (DSP) and ciguatera (HaUegraff
2004) Besides human health Hannful Algal Blooms (HABs) also give negative impact to
aquaculture tourism industries and to the ecology of the water system Various factors
have been associated to the occurrence of HABs phenomenon such as wind and currents
light temperature oxygen pH and nutrient uptake (Burkholder et ai 2006) Some species
were not bloom fonning but have potential toxin producers A few studies have done on
dinoflagellate in Sarawak River including Santubong River The study provides
infonnation on the species that available at Sarawak River but the composition of
dinoflagellates in the area of aquaculture is limited The aquaculture effluent might have an
influence to the species composition of dinoflagellates in that area and the potentially toxin
producer will gives negative impact not only to the river system but also to the consumer
who consume the seafood on that area
The objective of this study was to evaluate the occurrence of dinoflagellates species
composition at aquaculture area along Santubong River The composition of
dinoflagellates in Santubong River were also identified and documented This study could
provide an understanding on the documentation and all taxa of dinoflagellates presence on
the study area More over the possible toxic producers and bloom fonning dinoflagellates
can be identified
2
I
20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
l
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
I
20 LITERATURE REVIEW
Dinoflagellates are one of the most important components of phytoplankton in
marine and freshwater ecosystem (Spector 1984) The phytoplankton represent major
constituent of food webs and carry out photosynthesis to dependent aquatic ecosystem The
dinoflagellates are very abundant in all type of aquatic ecosystems and can be found living
as plankton or attached to sediments sand coral or on the other aquatic plants (Hoek et aI
1995)
21 Taxonomy of Dinoflagellates
Dinoflageliates are classified as Protists under the division Dinophyta in botanical
system and the order Dinoflagellida or Dinoflagellata in zoological systems (Hoppenrath amp
Saldarriaga 201 0) This is because some of the species are motile and heterotrophic while
other species have cell walls and are photosynthetic (Cabrini et aI 2010) Approximately
about 4500 species 550 genera taxonomic data of dinoflagellates have been described and
more than half of the species are in fossil fonn From 2000 living species more than 1700
are marine and about 220 are freshwater (Taylor et al 2007) Dinoflagellates show a great
range of fonns but most of this diversity can be resolved to five basic types (Taylor 1980)
including the hannful species
211 Dinoflagellates Morphology
Dinoflagellates are predominantly unicellular eukaryotic flagellated organisms that
posses both photosynthetic and non photosynthetic characters Their morphology is very
3
l
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
-----~------------~---------------------------
diverse They can be divided into two types depending on the point of insertion of the
flagella namely desmokont and dinokont (Taylor et aI 2007) Desmokont is a
dinoflagellate where the two flagella emerge at the cell apex While dinokont the flagella
emerge from the middle of the cell (Bellinger amp Sigee 2010) The two flageHa are inserted
ventrally one flagellum is longitudinal and housed in a sulcus The transverse flagellum
provides propulsion and the longitudinal flagellum provides direction (Tomas 1997) In
desmokonts the pole are directed towards the swimming direction is anterior the opposite
pole is posterior and the broad surfaces of the cell are lateral In dinokonts the side form
which the flagella arise is ventral the opposite surface is dorsal Some examples of
dinokont are Gymnodinium and Dinophysis
The other group desmokont is Prorocentrum (Taylor 1987) A dinoflagellate
lacking cellulose plates is said to be athecate and cell covered only by a membrane
(Cabrini et aI 2010) The plates may be thick or so thin that they cannot be seen with the
light microscope There is generally an inverse relationship between the number and the
thickness of the plates (Taylor 1980) The number shape and arrangement of these plates
form a distinctive geometry known as plate tabulation which is the main means for
classification and they are a most important taxonomic criterion (Taylor 1980)
wntnIlvlew dorsal view
Figure 1 Basic Anatomy of Thecate Dinokont Dinoflagellate Modified After Evitt 1985 (Macrae 2007)
4
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
Pusat Khidmat Maklumat Akademik UNlVERSm MALAYSIA SAKAWAK
Dinoflagellate usually has a cellulose cell wall perforated by many pores Most forms
have an equatorial groove that contains a ribbon flagellum This groove separates the
dinophytes cellulose cell wall into two portions the epicone and hypocone The number
arrangement and thickness of the thecal plates can vary significantly The cell surface
ornamentation can be observed to identify the species such as pores depressions spines
ridges and reticulations (Faust 2002) Spines wings and horns may decorate the cell
wall The thecate forms have covering composed of three parts epitheca (covering
epicone) girdle plate (covering the girdle) and hypotheca (covering the hypocone)
(Lebour 1925) Dinoflagellates are mostly having a single nucleus called the dinokaryon
The DNA is not organizing around nucleosomal histones instead forming fibrillar
chromosomes that are always condensed and divide through mitosis with an external
spindle (Spector 1984)
22 Distribution of Dinoflagellates
Dinoflagellates can be found with 90 in marine and 10 in freshwater in the aquatic
environment (Taylor et ai 2007) The distribution of dinoflagellates depends on wind and
currents light temperature oxygen pH and nutrient uptake Winds play an important role
in the development of freshwater dinoflagellate blooms whether in their vertical and
horizontal distribution (Taylor 1987) Dinoflagellates are commonly studied during their
motile planktonic stage cyst-forming dinoflagellates which are known from all oceanic
habitats Temperature can influence photosynthetic rates and division rates uptake and
respiration rates and cell size of the dinoflage1lates (Steidinger 1997)
5
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
tmiddot
23 Ecology and Growth of Dinoflagellates
Approximately 13-16 of living dinoflagellates produce a donnant resting cyst
(Head 1996) Cyst fonnation for dinoflagellates was basicaHy related to the fonnation of
seed to initiate red tides or Hannful Algal Blooms (HABs) existence approach due to
environmental stress resources for genetic recombination direct source of toxicity and
factor in bloom tennination (Anderson et ai 2003) Therefore dinoflagellates cysts
provide infonnation of mechanisms of spreading and reoccurrence of HAB (Sidharta et aI
2008)
Dinoflagellates are the second most abundant fonn of autotrophic life in the marine
ecosystem As such they are at the base of the food chain and provide food for herbivorous
zooplankton and sessile benthic suspension feeders They can be found either as free-
floating planktonic dinoflagellate to benthic habitats as attached with the bottom from
freshwater to estuaries and to hypersaline waters (Faust 2002) Dinoflagellates are
typically large-celled organisms such as Ceratium Peridinium and Peridiniopsis
Dinoflagellate can vertica]ly migrate up during the day when the light is strongest and
down at night (Rapport 1996) Dinoflagellates are the only photosynthetic organisms
capable of bioluminescence (Taylor 1987) Some species will act as parasite on other )
organisms and some species also support coral reef ecosystems through symbiotic
associations This type of dinoflagellate is called zooxanthellae
24 Harmful Algal Blooms (HABs)
Mass productions of phytoplankton are known as algal blooms Such blooms contain
- high concentrations of algal biomass HABs causing discoloration of water are commonly
6
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
known as red tides (Halle graeff 1995) Dinoflagellate species such as Dinophysis
Alexandrium and Prodinium can contaminate shellfish with toxins even at very low cell
concentration (Hallegraeff 2004) Harmful dinoflagellates are basically indistinguishable
from other dinoflagellates from the same habitats in their responses to temperature
salinity and light (Taylor 1987) Freshwater dinoflagellate has poor salinity tolerance
where estuarine and some marine species can tolerate a wide salinity range (Taylor 1987)
There are different types of harmful dinoflagellate blooms Most of the information is
adapted from Camacho et al (2006)
a) Species which produce basically harmless irritation conditions (odours andor
discoloration of water) in sheltered bays that causes oxygen depletion Example
Gonyaulax polygramma Gymnodinium sanguinum Noctiluca scintilans
b) Species that produce potent toxins that can find their way through the food chain
to humans causing a variety of gastrointestinal and neurological illnesses Some
of the species can contaminated the shellfish at very low cell concentrations
1 Diarrhetic Shellfish Poisoning (DSP) Mostly found in mussles
scallops clams and gastropod The main toxins were Okadaic acid
dinophysis toxins (DTXs) yessotoxins (YTXs) and pectenotoxins
(PTXs) Example Dinophysis acuta D acuminata D rotundata
Prorocentrum belizeanum P faustiae Plima
II Ciguatera Fish Poisoning (CFP)
Ciguatera fish poisoning is the most common marine toxin disease
worldwide The primary toxin involve is ciguatoxin and mostly been
contaminated if consume reef fish such as barracuda grouper and
snapper Example Gambierdiscus toxicus Ostreopsis
mascarenensis Prorocentrum sp
7
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
bull r
iii Paralytic Shellfish Poisoning (PSP) It is associated with saxitoxin
(STXs) Example Alexandrium acatenella A catenella A
cohorticula A fundyense A fraterculua A minutum A tamarense
Gymnodinium catenatum Pyrodinium bahamense var compressum
iv Neurotoxin Shellfish Poisoning (NSP) the toxins contain was
brevetoxins (PbTxs) Detected to who consume oyster clams
mussels cockles and whelks Example Karenina breve and K cf
breve from New Zealand
c) Species which are nontoxic to humans but harmful to fish and manne
invertebrates especially in intensive aquaculture systems The cells may cause
damage or clog the gills of these animals It is mostly attacked mussels and oyster
Example Gymnodinium mikimotoi
241 Impacts of HABs
HABs are natural phenomena but their occurrence geographic range and intensity
appear to have increased since the 1970s and their economic impact is larger now
compare to the past (Camacho et al 2006) The increased economic impact of HABs is
probably linked with the increased consumption of seafood and growth in coastal
populations (Anderson et al 2003) Symptoms occur generally as a consequence of
consumption of contaminated seafood and direct human exposure to HABs
8
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
30 MATERIALS AND METHODS
31 Study areas
The study was carried out at Santubong River between December 2011 and April 2012
Santubong River is an important river that located in Santubong Village which is about 35
km from Kuching City There is a lot of other village which depend on aquaculture
industries and also transportation in Santubong area Phytoplankton samples were collected
from three stations along the Santubong River The following months the sampling was
done with two other stations The stations were fish cages culture shrimp aquaculture
discharge and jetty of Fisheries Department The location of each site was coordinated
using Global Positioning System (GPS) All the locations were recorded in Table 1
Table 1 Sampjing sites (station) along the Santubong River
Station Location Coordinate
Jetty of Fisheries Department N 01 deg42 85 E 110deg19270
2 Fish Cage Culture
3 Shrimp aquaculture discharge
9
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
Figure 2 Maps of Santubong River
10
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
I shy
32 Phytoplankton Sampling and Water Quality Collection
Eighteen water samples were collected using Van Dom water sampler with 20llm
mesh size plankton net at the surface of the water column Three stations were selected
jetty of Fisheries Department shrimp fann discharge and fish cage culture On the first two
months of sampling December 2011 and January 2012 only one stations of sampling were
done The next sampling February to April 2012 were done in three stations with varies
tidal range The water samples are kept in the 1 L Whirl-Pak and kept in the cooler box Inshy
situ water quality namely temperature and pH using pH meter turbidity using turbidity
meter salinity was estimated using salinometer and dissolved oxygen (DO) using DO
meter The transparency of water column was detennined with a Secchi disk The depth of
the river is measured in every sampling using depth finder The means of temperature pH
salinity DO turbidity for each sampling station were obtain from three replicate readings
The water samples were transported to the laboratory and preserved with Lugol s
acidic solution Some fresh samples were observed under the compound microscope
immediately after field sampling Lugols solution is added at a concentration of 1 mL per
100 mL sample (APHA 1976) An advantage of Lugols solution is that flagellates
preserved with it retain their flagella (Hotzel amp Croome 1999) The water samples were let
to settle for at least 24 hour After that the upper layer of sediment samples were siphoned
off from the top layer to final volume of 100ml and 1ml aliquot was used for enumeration
(Bangheri et aI 2010) The nutrients of the water samples are refer to the previous study
done by Ling et al (2010)
11
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
I bull
33 Phytoplankton identification and enumeration
One ml aliquot from each sample was poured in the counting slide with coverslips
(24x20) and observed under compound microscope with 4X and lOX objectives (Liliana
2005) Phytoplankton were counted and identified to the lowest taxa under compound
microscope Magnus Live model equip with connection to the laptop to capture the picture
ofphytoplankton found
34 Statistical Data Analysis
Statistical analyses of data obtained from each station among months were determined
using SPSS version 20 for Windows Significant differences of these parameters at each
sampling station monthly were tested with one-way ANOV A with P005 using Tukey
statistical test (Liliana 2005)
)
12
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
40 RESULTS
41 Qualitative Phytoplankton Composition
The phytoplankton checklist and contribution of different taxonomic groups to the
total phytoplankton are presented in Table 2 A total of 89 taxa were identified in this
study From the list 8 genera consists of 17 species of dinoflagellates 69 species of
diatoms from 20 genera and blue-green algae Diatoms were most occurred taxa during
every months of sampling compare to dinoflagellates Family Chaetoceraceae from centric
diatom made up the higher number (2 genera 13 species) followed by family from
pennate diatom Bacillariaceae (6 genera 10 species)
=Dinoflagellate
~ Pennate Diatom
bull Centric Diatom
December January February March April
Month
Figure 3 Percentage Distribution of Phytoplankton
13
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
absent
four
42 Dinoflagellates Assemblages
The species composition of dinoflagellates was lower than diatoms throughout the study
period at all sampled locations Some were recorded throughout the months and some were
Seventeen species of planktonic dinoflagellates belonging to 8 genera were
recorded during this study They occurred throughout the period of study with distinct
monthly sampling Ceratium is the common and distributed genus in the study areas with
species of were identified namely Ceratium jurca Ceratium fosus Ceratium
trichoceros and Ceratium tripos Meanwhile the Dinophysis and Protoperidinium with
three species each The cell counts of C jurca were the highest during December 2011 and
those Protoperidinium on April 2012 off Santubong River
Many species of potentially toxic dinoflagellates were found in this study All of
them occurred in low cell numbers C jurca Cjusus Prorocentrum sigm 0 ides P micans
and Protoperidinium sp can be found in every months of the sampling location Individual
oPyrophacus Pyrocystis lunula and Dmile were less detected at all locations
)
14
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
i ll50 iI e ~
18
16
14 tj
12I 0 0 IOc- 0
8 v 0
~ 6 Z
4
2
0
tshy__________________________________________LI_______
I middot
J------------------------------------li --shy I r--shy
1---------------------------------lJ-e-shy ~ middot 1
-+
i I
r j ~ J= I~ ~
I
December January February March April
-
-
lr I
o Ceratiaceae
E3 Dinophysiaceae
I Gymnodiniaceae
11 Prorocentraceae
El Protoperidiniaceae
II I Pyrocystaceae
~ Pyrophacaceae
Month
Figure 4 Number of Dinoflagellates Species identified in Monthly Sampling
400 ~-------------------------------
350
300
250
200
100 II Number of Individual
50 o
IV b-IV IV IV IV IVb- b- b- b- bshyilJ bull IV IV ~
IV IV IV ~1gt 1gt 1gt~~ iY~ IVlt iY~ ~1gt~o~ ~o ZJ~ o~ampv ~oQ-lty ()4-$i ~o ~o~ ~ ~4
~lto
Family of Dinoflagellates
Figure 5 Number of dinoflagellates individual per ml along Santubong River
15
