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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

http://aem.asm.org/cgi/content/full/66/1/439/F1

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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