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Master's Dissertation
DOI
https://doi.org/10.11606/D.18.2018.tde-01092020-170215
Document
Author
Full name
Janaína Fagundes Malta
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Carlos, 2018
Supervisor
Committee
Cunha, Davi Gasparini Fernandes (President)
Mota Filho, Cesar Rossas
Seleghim, Mirna Helena Regali
Title in Portuguese
Degradação biológica de microcistina em laboratório: subsídios para o tratamento avançado de águas de abastecimento
Keywords in Portuguese
16S rRNA
Biodegradação
Cianotoxinas
Comamonadaceae
Microcistina
Tratamento Avançado de águas de abastecimento
Abstract in Portuguese
Frequentemente encontrada nas florações de cianobactérias tóxicas em mananciais e reservatórios de água eutrofizados, a microcistina-LR (MIC-LR) é uma hepatotoxina persistente e de difícil remoção em Estações de Tratamento de Água (ETAs). O tratamento biológico combinado com o processo convencional de tratamento tem demonstrado resultados satisfatórios na remoção de MIC-LR, por meio da utilização de microrganismos na biodegradação da cianotoxina. Esta pesquisa avaliou, em diversos experimentos laboratoriais com duração de 10 dias, a biodegradação de MIC-LR por diferentes inóculos, como culturas purificadas de Sphingomonas sp. e Brevundimonas sp., comunidade bacteriana autóctone isolada de um sistema de abastecimento de água e do próprio extrato da toxina. Foram testadas diferentes condições de concentração inicial (C0) de MIC-LR, temperatura (T) e diferentes volumes de microrganismos inoculados (v/v). As análises das concentrações residuais da toxina foram realizadas pelo método Elisa (Enzyme-Linked Immunosorbent Assay) e a análise filogenética dos microrganismos, pelo sequenciamento do gene 16S rRNA. Os resultados obtidos foram estatisticamente significativos para C0, T e inóculos avaliados, nos quais influenciaram na redução substancial da toxina. Em C0 = 100 µg.L-1 , as menores concentrações residuais foram observadas pela comunidade bacteriana autóctone presente na ETA, que juntas degradaram mais de 90% da toxina, quando comparado as culturas purificadas de Sphingomonas sp. e Brevundimonas sp., que reduziram em até 60% a C0. Quando expostos a C0 = 20 µg.L-1 , os resultados demonstraram redução de 96% da concentração da toxina pela (SF); 90% na entrada e saída do filtro rápido (EF e SF) e 80%, pelos inóculos de culturas purificadas de Sphingomonas sp. e Brevundimonas sp. na concentração de 10% (v/v), nas T = 22°C. Este é, possivelmente, o primeiro relato no Brasil da capacidade de cepas bacterianas Brevundimonas sp. em degradar MIC-LR. O acréscimo de T foi acompanhado pelos melhores resultados obtidos nesta pesquisa, com a biodegradação de 99% e 97% da toxina (C0 = 100 µg.L-1) pelos inóculos da SF e comunidade bacteriana autóctone do extrato de MIC-LR. Um total de 464 OTU (Operational Taxonomic Unit) foi obtido da comunidade bacteriana autóctone presente no extrato da toxina, EF e SF, com predominância de quatro filos: Proteobacteria, Actinobacteria, Bacteroidetes e Firmicutes (abundâncias relativas de 51,7, 13,4, 6,5 e 2,8%, respectivamente). Além disso, foi verificada a presença da família Comamonadacea em todas as amostras analisadas, sugerindo-se a participação de cepas dessa família na biodegradação da toxina. Esta pesquisa demonstrou, por meio da utilização de microrganismos e diferentes fatores bióticos e abióticos, resultados promissores ao desenvolvimento dos processos biológicos como etapa adicional ao tratamento avançado de águas contaminadas por MIC-LR.
Title in English
Biological degradation of microcystin in laboratory conditions: contributions to drinking water advanced treatment
Keywords in English
16S rRNA
Biodegradation
Comamonadacea
Cyanotoxins
Drinking water advanced treatment
Microcystin
Abstract in English
Often found in toxic cyanobacterial blooms in eutrophic water sources and reservoirs, microcystin-LR (MIC-LR) is a persistent hepatotoxin that is difficult to be removed in drinking water treatment plants (DWTPs). Biological treatment combined with conventional treatment process has shown satisfactory results in the removal of MIC-LR, through the use of microorganisms to biodegrade cyanotoxin. This study evaluated the biodegradation of MIC-LR by different inocula, such as purified cultures of Sphingomonas sp. and Brevundimonas sp., bacterial community isolated from a water supply system and the toxin extract itself. Several laboratory experiments were conducted with a duration of 10 days each. The biodegradation tests were carried out considering different initial concentrations (C0) of MIC-LR, temperature (T) and different volumes of inoculated microorganisms (v/v). Residual toxin concentrations analyses were performed by ELISA (Enzyme-Linked Immunosorbent Assay) and for phylogenetic analysis of bacterial communities 16S rRNA gene sequencing was applied. The results were statistically significant for C0, T and the evaluated inocula, these factors influenced the substantial toxin reduction. With C0 = 100 µg.L-1, the lowest residual concentrations were observed by the bacterial community present in the DWTP, which degraded more than 90% of the toxin when compared to the purified cultures of Sphingomonas sp. and Brevundimonas sp., which reduced C0 up to 60%. When exposed to C0 = 20 µg.L-1, the results demonstrated a 96% reduction in toxin concentration by the bacterial community posfiltration of the rapid sand filter (PF); 90% at the prefiltration and posfiltration of the rapid sand filter (PR and PF) and 80% by the purified cultures inocula of Sphingomonas sp. and Brevundimonas sp. at the concentration of 10% (v/v) at T = 22°C. This is possibly the first report in Brazil of the capacity of Brevundimonas sp. bacterial strains in degrading MIC-LR. The addition of T was followed by the best results obtained in this research, with the toxin biodegradation of 99% and 97% (C0 = 100 µg.L-1) by PF inoculums and bacterial community from MIC-LR extract. A total of 464 OUT (Operational Taxonomic Unit) was obtained from the bacterial community present in the toxin extract, PR and PF, with four predominant phyla: Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes (relative abundances of 51.7, 13.4, 6.5 and 2.8%, respectively). In addition, the presence of the Comamonadacea family was verified in all analyzed samples, suggesting the participation of this family strains in the toxin biodegradation. This research demonstrated, by using microorganisms and different biotic and abiotic factors, promising results to the development of biological processes as an additional step to the advanced treatment of water contaminated with MIC-LR.
 
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Publishing Date
2021-06-22
 
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