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Indigenous
Hydrocarbonoclastic Bacteriain Jakarta Bay and SeribuIsland Waters
Yeti Darmayati, Shigeaki Harayama*, AtsushiYamazoe*, Ariani Hatmanti, Sulistiani,
Ruyitno and Djoko Hadi Kunarso
(LIPI,Indonesia and NITE, Japan)
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Preliminary study
Jakarta Bay polluted byoil (14 78 ppm)
Potential sources of oilpollution in Seribu islands
: activities in Jakarta Bayand Seribu islands
Oil pollution controlshould be done
Bioremediation :1st Understand ecologicalprocess
Study Purpose :
To isolate and identifyindigenoushidrocarbonoclastic bacteriain Jakarta Bay and
surroundings
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METHOD AND MATERIALS
Location
106.5E 106.6E 106.7E 106.8E 106.9E 107E
-6.15S
-6.10S
-6.05S
-6.00S
-5.95S
-5.90S
-5.85S
-5.80S
1
2
3
4
5
J A K A R T A B A Y
.Rambut Island
.Pari Island
.Pramuka Island
T A N G E R A N G
J A K A R T A
N
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Enrichment and Isolation
Oil degrader
Sublimation method
EnrichmentPAHs degrader
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Poly-aromatic Hydrocarbons
Dibenzothiophene (100oC, 5 min)
Phenanthrene (100oC, 5 min)
Pyrene (130oC, 10 min)
Naftalene (75oC, 1 min)
Fluoranthene (95oC, 10 min)
Fluorene (100oC, 5 min)
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16S rRNA gene sequencedetermination
Step:Collect cell from pure cultures
Extract DNA
Amplify 16S rRNA genes with universalprimers
Purify the PCR products
Sequence the product
Detect and discard putative chimera
Align the sequences to database
Primer27 F: 5-AGRGTTTGATCMTGGCTCAG-31492R: 5-TACGGYTACCTTGTTAYGACTT-3
PCR cycle95C 3 min95C 1 sec55C 1 min72C 2 min72C 7 min4C 5 min ---
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RESULT
Oil polluted waters Unpolluted waters
Visually, the fastest degradation rate may occurred inMarina Port.
131 isolat hydrocarbonoclastic bacteria consist of 65
different strains.
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Clone library result : 171 from Pari island waters
139 from Jakarta Bay
17
29
17
15
9
0
5
10
15
20
25
30
Marina Muara baruoffshore
Rambut Island Pari Island Pramuka Island
Strain Number in Different Sites of Jakarta Bay and
Surrounding Waters
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Hydrocarbonoclastic bacteria: Achromobacter,Acinetobacter, Alcaligenes, Arthrobacter, Bacillus,Brevibacterium, Corynebacterium, Flavobacterium,
Nocardia, pseudomonasand Vibrio
Indonesia Pseudomonas cepacia and P. gladioli(East
Kalimantan, 2003)
Achromobacter putrefaciens, Acinetobacterhaemolyticusand Vibrio alginolyticus( Javasea, 2004)
Acinetobacter, Arhtrobacter, MicrococcusandBacillus( Malaka straits, 1998)
Alcanivorax, Marinobacter, BacillusandAchromobacter (Jakarta Bay and SeribuIsland,2005)
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Alcanivoraxand Marinobacter:cosmopolitan
Bacillus : clean water
Achromobacter : oil polluted area
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12.98
3.82
8.40
41.98
7.63
16.03
9.16
DBT FLU FRT OIL NAF PHE PYR
Alcanivorax,Bacillus
Sphingopixis,Bordetella
AlcanivoraxKoordiimonas
Bordetella, AlcanivoraxCytophoga, Alcanivorax
Tistrella,Alcanivorax
Percentage of Isolates Number on Degrading Oil andPAH Capability
Brevibacterium
Marinobacter
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Hidrocarbonoclastic bacteria indifferent sites
0
5
10
15
20
25
Water Sediment Water Sediment Water Sediment
Marina Muara Baru offshore Rambut Island Pari Island Pramuka
Island
DBT
PHE
FLU
NAF
PYR
FRT
Oil
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Conclusion
Oil pollution level may not impact the numberof strain hydrocarbonoclastic bacteria isolated.
Oil composition may impact communitystructure of hydrocarbonoclastic bacteria
Alcanivoraxspp. is not only important oil andPAH-degrader bacteria in temperate area, butits may also in tropical areas.
Alcanivorax, Marinobacter, Achromobacterand
Bacillusare common hydrocarbonoclasticgenera bacteria in Jakarta Bay and itssurrounding waters.
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Acknowledgment
National Institute of TechnologyEvaluation, Japan
NEDO, Japan
Indonesian Institute of Sciences,Indonesia
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THANK YOU FOR YOUR
ATTENTION