Isolation and 16S rRNA characterization of culturable bacteria derived from fire coral Millepora intricata in Morela Coastal Waters, Maluku, Indonesia
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Abstract
Abstract. Agung MUK, Khairunnisa N, Astuty S, Lewaru MW, Mulyani Y. 2021. Isolation and 16S rRNA characterization of culturable bacteria derived from fire coral Millepora intricata in Morela Coastal Waters, Maluku, Indonesia. Indo Pac J Ocean Life 4: 1-7. Millepora, well recognized as fire coral, has nematocytes that contain toxins as self-defense, allowing the associated bacteria to have a high level of adaptation to survive in this extreme environment. This research aims to isolate and characterize fire corals Millepora intricata-associated bacteria obtained from Morela coastal waters, Maluku, Indonesia using classical culture method. Sampling of M. intricata was carried out in August 2017 from the depth of 2-3 meters. The bacteria were grown in sea water-Nutrient Agar (NA) medium, streak plate method then used until pure isolates were obtained. Molecular identification of isolates was performed using 16S rRNA gene marker. The results indicated that nine culturable pure isolates were successfully obtained from M. intricata. Three isolates (MI.P1.B; MI.P2.C; and MI.P2.D2.K) then further proceed into molecular identification and sequencing. Alignment result based on the 16S rRNA databases repositored in the genebank (NCBI) showed that isolate MI P1.B has similarity to registered sequence of Pseudoalteromonas arabiensis strain k53 (NCBI acc no. NR_113220.1) with a sequence identity of 98%. Isolate MI.P2.C has a close relationship (98%) to registered sequence of Halomonas aquamarina strain DSM 30161 (NCBI acc no. NR_042063.1). Meanwhile, isolate MI.P2.D2.K has similarity to registered sequence of Halomonas zhanjiangensis strain JSM 078169 (NCBI acc no. NR_104283.1) with a sequence identity of 99%.
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Alvarado CI, J Alejandro G, Manuel BA, Alejandra R, Andres F, Edgar PHC. 2007. Biochemical and pharmacological characterization of toxins obtained from the fire coral Millepora complanata. Comp Biochem and Physiol Part C 146: 511-518.
Arahal DR, Wolfgang L, Karl HS, Antonio V. 2002. Phylogeny of the family Halomonadaceae based on 23S and 16S rDNA sequences analyses. Int J of Syst and Evol Microbiol 52: 241-249.
Atencio LA, Francesco DG, Giannina OY, Ronnie G, Hector MG, Imke S, Luis CM, Marcelino G. 2018. Antimicrobial-producing Pseudoalteromonas from the marine environment of Panama shows a high hylogenetic diversity and clonal structure. J Basic Microbiol 58: 747-769 doi: 10.1002/jobm.201800087
Cappuccino JG, Welsh C. 2018. Microbiology a Laboratory Manual. Eleventh Edition. Pearson, England.
Chen YG, Yu-Qin Z, Heng-Yu H, Hans-Peter K, Shu-Kun T, Ke H, Qi-Hui C, Xiao-Long C, Wen-Jun L. 2009. Halomonas zhanjiangensis sp. nov., a halophilic bacterium isolated from a sea urchin. Int J of Syst and Evol Microbiol 59: 2888-2893 doi: 10.1099/ijs.0.010173-0
Coenye T, Vandamme P. 2003. Diversity and Significance of Burkholderia Species Occupying Diverse Ecological Niches. Environmental Microbiology. Vol 5 (9) : 719-729 doi:10.1046/j.1462-2920.2003.00471.x
Devereux, R., Wilkinson SS. 2004. Amplification of Ribosomal RNA Secquences. Kluwer Academic Publisher : Netherlands
Dobson SJ, Franzmann PD. 1996. Unification of the genera Deleya (Baumann et al. 1983), Halomonas (Vreeland et al. 1980), and Halovibrio (Fendrich 1988) and the species Paracoccus halodenitrificans (Robinson and Gibbons 1952) into a singel genus, Halomonas, and placement of the genus Zymobacter in the family Halomonadaceae. Int J of Syst Bacteriol 46(2): 550-558.
Espiritu RA. 2016. Preliminary Bioactivity Screening of Dichloromethane and Methanol Extracts from the Fire Coral Millepora dichotoma, Der Pharmacia Lettre, 8 (12) : 105-109
Espiritu RA, Raymond SM, Consolacion YR. 2016. Chemical constituents of Millepora dichotoma. Der Pharm Chem 8(3): 245-248 ISSN 0975-413X CODEN (USA): PCHHAX.
Fatkhurrochman. 2017. Aktivitas Antibakteri dari Bakteri Asosiasi Karang Api (Millepora sp.) Terhadap Bakteri Patogen pada Karang yang Terkena Black Band Disease (BBD). [Undergraduate Thesis/ Unpublished]. Universitas Padjadjaran, Jatinangor [Indonesian]
Frank JA, Claudia IR, Shobha S, Jon SW, Brenda AW, Gary JO. 2008. Critical evaluation of two primers commonly used for amplification of bacterial 16S rRNA genes. Appl and Environ Microbiol 2461-2470.
Hillis DM, James JB. 1993. An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Syst Biol 42(2): 182-192
Hutchings P, Saenger P. 1987. Ecology of Mangroves. University of Queensland Press, St Lucia, Qld.
Integrated Taxonomic Information System (ITIS). Available: http://www.itis.gov. [28 September 2018]
Lane DJ. 1991. 16S/23S rRNA Sequencing. In: Stackebrandt, E., Goodfellow, M., editors. Nucleic Acid Techniques in Bacterial Systematics. Wiley & Sons, New York p. 115-76
Leboffe MJ, Pierce BE. 2012. Microbiology Laboratory Theory & Application: Brief. Second Edition. Morton Publishing Company, Englewood, 642 pp
Li S, Dennis KP, Hani D. 1996. Phylogenetic tree construction using markov chain monte carlo. https://pdfs.semanticscholar.org/7cfb/3b0dfde70d1b25c97d888a908e5a637b678b.pdf
Madigan MT,Martinko JM, Bender KS, Buckley DH, Stahl DA. 2015. Brock Biology of Microorganisms. 14th ed. Pearson, England.
Mata JA, Jose MC, Emilia Q, Victoria B. 2002. A detailed phenotypic characterisation of the type strains of Halomonas species. Syst Appl Microbiol 25: 360-375.
Matsuyama H, Hideki M, Hirokazu K, Yoshihisa K, Masafumi M, Isao Y. 2013. Pseudoalteromonas arabiensis sp. nov., a marine polysaccharide-producing bacterium. Int J of Syst and Evol Microbiol 63: 1805-1809 doi: 10.1099/ijs.0.043604-0.
Moats W.E. 1992. Fire Coral Envenomation, Journal of Wilderness Medicine, 3, p 284-287
Morrow KM, Moss AG, Chadwick NE, Liles MR. 2012. Bacterial associates of two Caribbean coral species reveal species-specific distribution and geographic variability. Appl Environ Microbiol. 78: 6438–6449. doi:10.1128/AEM.01162-12
Nybakken JW. 1992. Biologi Laut Suatu Pendekatan Ekologis. Alih Bahasa: HM Eidman, Koesoebiono, DG Bengen, M Hutomo & Sukardjo. PT. Gramedia, Jakarta 459 hal.
Pelczar MJ, Chan ECS. 1986. Dasar Dasar Mikrobiologi Jilid 1. Penerjemah: Hadioetomo RS, T Imas, SS Tjitrosomo, SL Angka. UI Press, Jakarta.
Razak, TB, Hoeksema BW. 2003. The Hydrocoral genus Millepora in Indonesia. Zool. Verh. Leiden 345 : 313-336
Radjasa, OK, Sabdono A. 200. Screening of Secondary Metabolite-Producing Bacteria Associated with Corals Using 16S rDNA-Based Approach, Journal of Coastal Development Vol. 7 (1) : 11-19
Sidharta BR. 2000. Pengantar Mikrobiologi Kelautan. Universitas Atmajaya, Yogyakarta 122 hal.
Suantika G, Pingkan A, Dea IA, Zarah FK. 2013. The use of indigenous probiotic Halomonas Aquamarina and Shewanella algae for White Shrimp (Littopenaeus vannamei Boone) hatchery productivity in zero water discharge system. J Aquac Res Dev 4: 194 DOI: 10.4172/2155-9546.1000194.
Suharsono. 2008. Jenis-jenis Karang di Indonesia. Coremap Program. LIPI Press, Jakarta 344 hal.
Veron J. 2000. Corals of the World Vol. 3. Australian Institute of Marine Science, Townsville Australia.
Woese CR, Fox GE. 1977. Phylogenetic Structure of the Prokaryotic Domain : The Primary Kingdoms. Proc. Natl. Acad.Sci.USA. Vol. 74 (11) : 5088-5090
Wu Y, Hong C, Lin X, Xiong-Bin J, Chun-Sheng W, Xue-Wei X. 2017. Physiological and genomic features of a novel violacein-producing bacterium isolated from surface seawater. PloS ONE 12(6): e0179997.
Zhou Y, Rui L, Xiao-Yang G, Alla L, James H, Matthew H, Elizabeth L, Marcel H, Amrita P, Natalia NI, Manfred R, Konstantinos M, Brian JT, Victor M, Tanja W, Hans-Peter K, Nikos CK, Wen-Jun L. 2014. High quality draft genome sequence of the slightly halophilic bacterium Halomonas zhanjiangensis type strain JSM 078169T (DSM 21076T) from a Sea Urchin in southern china. Stand in Genom Sci 9: 1020-1030 DOI: 10.4056/sigs.5449586.