Microbial community of a volcanic mudspring in the Philippines as revealed by 16S rDNA sequence analysis and fluorescence in situ hybridization |
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Authors: | Nacita B Lantican Ma Genaleen Q Diaz J Jason L Cantera III" target="_blank">Francis L de los ReyesIII Asuncion K Raymundo |
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Institution: | 1.Institute of Biological Sciences, College of Arts and Sciences,University of the Philippines Los Ba?os, College,Los Ba?os,Philippines;2.Environmental Sciences Department,University of California,Riverside,USA;3.Civil, Construction, and Environmental Engineering,North Carolina State University,Raleigh,USA;4.Dean’s Office, College of Arts and Sciences,University of the Philippines Los Ba?os, College,Los Ba?os,Philippines |
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Abstract: | Mt. Makiling Mudspring in Laguna, Philippines is a thermophilic, acidophilic environment that previously has been shown to
harbor novel microorganisms. We assessed the microbial community that exists at this volcanic mudspring using 16S rRNA-based
approaches. DNA was extracted from solfataric soils and sediments taken from Mudspring. The 16S rDNA was PCR amplified using
universal (519F-1392R) and archaeal-specific (23FPL-1391R) primer pairs, cloned, and sequenced. Phylogenetic analysis of the
cloned 16S rDNA showed that eleven clones clustered with, and therefore related to Sulfolobus tokodaii 7 and two clones clustered with S. solfataricu, S. shibatae and S. islandicus. Three clone sequences were related to those found in thermophilic chalcopyrite (CuFeS2), a copper sulfuric ore from bioleaching reactors. One clone had low similarity (95% identity) with uncultured archaeon clone
KOZ184. Fluorescence in situ hybridization (FISH) analysis revealed that about 71% of the microbial community present in the
Mudspring belong to domain Archaea of which 63% were Crenarchaeota and 8% were Euryarchaeota. Seventeen percent (17%) of the
population consisted of bacteria as indicated by the positive hybridization with the BACT338 probe, and the remaining 12%
are unidentified. This study is the first attempt to use molecular techniques in any environment in the Philippines. |
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