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ALMEIDA, B.C. Bacterial diversity in seawater samples at São Sebastião Channel. 2009. 198 f. Ph.D. Thesis (Microbiology) Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, 2009. The marine environment is characterized by typical ecological setting, as high pressure, salinity, temperature, light, and others and by sustains a vast bacterial diversity. The study of seawater bacterial diversity is so much important to understand the structure and the patterns distribution of the communities and this could be done combining conventional and contemporary methodologies. Determine the concentration of viable heterotrophic bacteria, total bacterial communities, and the bacterial identification were the objectives of this study. During 20 consecutive months (August 2005 to March 2007) marine viable bacteria were counting and isolated from two points of collection at São Sebastião Channel using plate count and marine agar. The concentration of marine viable bacteria varied from 2.0 x 10 1 to 3.4 x 10 3 CFU/mL in 40 seawater samples collected and 295 isolates were obtained. Preliminary viable bacterial characterization was done using Gram staining and only the Phylum Proteobacteria was identified by 16S rRNA partial sequencing. The molecular genotyping was done by BOX-PCR e ERIC-PCR. One hundred and sixty eight marine viable bacteria were sequenced and 86.9% of them belonging to Vibrio genus, however the other were homologous with Photobacterium, Pseudomonas, Aeromonas, Alteromonas, Pseudoalteromonas, Shewanella, Alcaligenes, Yangia, Paracoccus and Thalassobius genus. The concentration of proteobacteria communities was verified using the fluorescence in situ hybridization (FISH) methodology for the Alpha-, Beta-, Gammaand Deltaproteobacteria classes and the frequencies were 9.5%, 10.2%, 8.1% e 6.8%, respectively when related to total cells count using DAPI staining that varied from 7.0 x 10 6 to 2.3 x 10 7 cells/mL. Although the Betaproteobacteria showed high frequency, the values between the Alpha-, Betaand Gammaproteobacteria classes were not different statistically. The total community was identified by construction of three bacterial libraries of 16S rDNA gene clones. The library 1, corresponding to seawater sample collected in November 2006 was compound by 98 clones. The Library 2, month February 2006 included 93 clones and the library 3, February 2007, was represented by 94 clones. The diversity was defined in terms of phylotypes or operational taxonomic units (OTUs). The phyla Proteobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, Fusobacteria, Verrucomicrobia and Chloroflexi were identified in the three libraries of seawater samples. The diversity indices and richness were determined using Chao1, ACE and Shannon index, however, the clones number sequenced were not enough to determine the bacterial richness at São Sebastião Channel.

Diversidade de bactérias em amostras de água do mar no canal de São Sebastião