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Accumulation of Tributyltin (TBT) and Intersex Occurrence as TBT-contaminated indicator on

the marine snail Littoraria scabra(Gastropoda: Mollusca)

Laboratory of Toxicology and Marine PharmaceuticsFaculty of Fisheries & Marine Sciences

Sam Ratulangi University, Manado, North Sulawesi, Indonesia

By Markus T. Lasut

TBT-contaminated indicator

Presented in:

International Conference & Workshop XI of “Tropical Marine Mollusc Programme”, sponsored by DANIDA at Kodaikal, Rameswaran and

Tuticorin-Tamilnadu, India. 28 September – 8 Oktober, 2000

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The role of molluscs in control & monitoring programme of marine environmental quality Toxic Substances

Discharge as residues

Introduced in purposes

MARINE ENVIRONMENTEffects:•Biological,•Instability,•Reduce species

BIVALVIA•Shell thickening

GASTROPODS•Imposex, Intersex

CONTROL & MONITORING PROGRAMME

OrganotinTributyltin

(TBT)•Measurement on the concentration•Biomonitor using

MOLLUSCS

Accumulation

As antifoulingpaints

INTRODUCTION

INTERSEX

MOLLUSCS

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INTERSEX:

1. Pathological alteration in the oviduct of littorinids, and2. Replacement of female by male organs

Has been proven (together with imposex) as a highly sensitive biomarker for the determination of the degree of environmental organotin and especially tributyltin (TBT)

Atlantic, North Sea, Baltic Sea

Littorina littorea

(continued)INTRODUCTION

Littorina littorea

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Littorina littorea

Littorina scabraspecies complex

Reid (1986) divided and reallocated this species into 16 species of the genus Littoraria

1 species of the genus Littoraria is L. scabra

(continued)INTRODUCTION

Littoraria scabra

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OBJECTIVES

1. Accumulation of TBTto understand two different pathways of TBT accumulation (via TBT-contaminated water & TBT-contaminated sediment and food)

2. Intersex occurrenceto measure and compare intersex stage between two different locations

Residues of TBT accumulated in the body of the animals were measured qualitatively using Tin-Layer Chromatography technique

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MATERIALS AND METHODS

1. Accumulation of TBTLittoraria scabra• collected at intertidal area• 15 animals per treatment• Shell length of 17.46 – 20.16 mm

TBT• Pure TBT + tween 80

a. Accumulation via TBT-contaminated water

TBT : 1, 2 and 3 ppb + control (treatment)Seawater : vol. 250 ml, 22.6 ppt, 28-30 oC, pH 7.50-7.70

b. Accumulation via TBT-contaminated sediment and food

TBT : 1, 2 and 3 ppb + control (treatment)Seawater : vol. 250 ml, 22.6 ppt, 28-30 oC, pH 7.50-7.70

• Daily observation was carried out on each treatment along 15 days

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MATERIALS AND METHODS

2. Intersex Occurrence Measurement on Intersex Stage has been done by calculating Intersex Index (ISI) as the mean value of the Intersex Stage in a population

ISI > 1.0 indicate that at least some of the females in the sample are sterilized due to intersex development

ISI = Intersex Stage values of all females sampled / number of females

Two different locations were chosen:• Amurang Bay : indicated by small scale/traditional harbour• Bitung Coastal Waters : indicated by large scale harbour

(this location then is divided by 5 sub-locations (A, B, C, D and E) to see the gradient of TBT contamination)

(continued)

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MATERIALS AND METHODS

1. Intersex Occurrence

Littoraria scabra• Shell length : 17.46 – 20.16 mm• Sampled during low-tide at the area• Anesthetic in 7% MgCl2 for maximum relaxation• Observation under microscope

Tin-layer Chromatography techniquewas applied to confirmed residues of TBT accumulated in the body of animals

(continued)

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RESULTS

yes (before 9 days)

yes, very rough

produce mucus

Behaviour : yes (before 9days)

Water condition: very rough

Other : produce mucus

3 ppb TBT

yes (before 9 days)

yes, rough

produce mucus

Behaviour : yes (before 9days)

Water condition: yes, rough

Other : produce mucus

2 ppb TBT

no (after 9 days)

no

--

Behaviour : no (after 9 days)

Water condition: no

Other : --

1 ppb TBT

no

no

--

Behaviour : no

Water condition: no

Other : --

Control

TBT-contaminated sediment & food

TBT-contaminated waterTreatments

Pathways

Visual Observation on L. scabra contaminated to TBT at different pathways

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RESULTS

Qualitative measurement of TBT residue accumulated in the body (soft part) of L. scabra

yesyesno

TBT-contaminated sediment & food

yesyesno

TBT-contaminated water

321ways

TBT Concentration (ppb)Path-

(continued)

no : undetected residuesyes : detected residues

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0.27929E

0.501728D

0.932231C

0.761330B

0.15526A

Bitung Coastal Water (BCW)

0.08536Amurang Bay

ISIIntersex

(%)

Number of Samples (Female)

Locations

Intersex Determination in Littoraria scabra

RESULTS (continued)

ISI > 1.0 indicate that at least some of the females in the sample are sterilized due to intersex development

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0

0.2

0.4

0.6

0.8

1

Am. Bay A B C D E

Locations

ISIValues

Intersex Intensity Comparison

Qualitative residue of TBT in body of L. scabra

noyesyesyesnono

EDCBABay

Bitung Coastal WatersAmurang

RESULTS (continued)

no : undetected residues, yes : detected residues

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DISCUSSION

Organotin(TBT)

Availability•Water•Sediment & foodBody system of

Littoraria scabra

Effects

Accumulation•Uptake•diffusions

Sublethal:•Spend energy•Intersex•behavioral

Lethal:•mortality

Depuration

If Accumulation > depuration = Effects

Contamination

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CONCLUSION

TBT compound was accumulated in both pathways (via TBT- contaminated water and TBT-contaminated Sediment & food) by the marine snail Littoraria scabra, start from 2 ppb of experimental concentration

Bitung Coastal Waters (BCW) is more pollute by TBT than Amurang Bay (AB), although it is not in dangerous stage of intersex

Intersex is found as a sensitive indicator for TBT contamination in low concentration Intersex occurrence in marine snail Littoraria scabra can be used in Control & Monitoring Program for TBT contamination

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ACKNOWLEDGEMENT

I am greatly indebted to the DANIDA sponsored Tropical Marine Mollusc Programme (TMMP) for the possibility to present this paper at the 11th TMMP Conference/Workshop at Kodaikanal, Tamilnadu, India, to Dr. Jorgen Hylleberg for his kindly help and advice.

I am particularly grateful to the Organizing Committee,Dr. P. Edward and Dr. A. Murugan for their warm welcome and hospitality during the congress.

“Thank you”