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BURBANO-ROSERO, E. M. Frequency and diversity of somatic coliphages isolated from seawater, plankton and bivalves samples from Baixada Santista, Canal de São Sebastião and Ubatuba. 2009. 155 p. Ph. D. Thesis (Science) Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, 2009. The somatic coliphages (SC) were indicated as the better indicator for fecal pollution in the microbiological quality of the marine ecosystem. However, in spite of the abundance of tailed coliphages, very little is known about its diversity. In this research, it was obtained the abundance of somatic coliphages in seawater (73), plankton (46) and bivalves (11) samples collected from 3 coastal regions of São Paulo: Santos, São Sebastiâo and Ubatuba. Somatic coliphages counts were correlated with marine viable bacteria (p=0.0001), thermotolerant coliforms (p=0.0001), E. coli (p=0.0001) and intestinal enterococci (r=0.247, p=0.035). Regarding the physico-chemical parameters, it was found a negative correlation with temperature (p=0004). In Ubatuba, the seawater samples no showed somatic coliphages. However, there were low concentration in plankton samples (<1 to 10 PFU/g) and bivalves (<1 to 14 PFU/g). In São Sebastião, the counts in seawater samples ranged from <1 to 15 PFU/100 mL, in plankton from <1 to 4.2 x10 2 PFU/g and in bivalves from <1 to 2 PFU/g. In seawater samples collected from Santos region were found the highest counts of coliphages (<1 to 3.4 x10 3 PFU/100 mL) compared to the plankton (<1 to 4.7 X10 2 PFU/g) and bivalves samples ( <1 to 22 PFU/g). In all, three regions, four morphotypes were observed: A1 (3%), B1 (63%), C1 (21%) and D1 (13%). The frequency of taxonomic SC families found in seawater and plankton samples were: Siphoviridae (50% and 65.8%), Podoviridae (36% and 15.8%), Microviridae (9% and 15.8%), and Myoviridae (5%, 2.6%), respectively. In bivalves, only the family Siphoviridae was observed. In all cases, especially those with flexible and curled tails, Siphoviridae was more abundant. The sixty two coliphages isolated were DNA and were characterized by RFLP technique using the following restriction enzymes: BamHI, BglII, DraI, EcoRI, EcoRV, HindIII, HinfI, HaeIII and RsaI. Better efficiency of digestion was obtained with: DraI (54.8%), EcoRI (40.3%), EcoRV (19.4%) and BgIII (14.5%). Among the rep-PCR techniques used verified that the BOX-PCR allowed a better molecular characterization. However, none of the methods used was useful to group according to sample type, source of isolation and/or taxonomic classification. It was also investigated the presence of the genes encoding CT, TCP, LT and ST toxins, and 9,6% of coliphages contained genes codifying for thermostable toxin (ST) and/or thermolabil toxin (LT). This study is identifying the coliphages as microbial hazard in the coastal region that could affect the animal and human health, giving support to later studies for microbial risk assessment of marine ecosystem.

Frequência e diversidade de colifagos somáticos isolados de amostras de água do mar, plâncton e bivalves da baixada santista, canal de São Sebastião e Ubatuba.