ABSTRACT
Para minimizar os problemas relacionados à ocorrência de cianobactéria em águas destinadas ao consumo humano há necessidade de se realizar estudos de alternativas técnicas de tratamento com destaque aos biofilmes com potencial de degradação de microcistinas (MC). O presente trabalho teve como objetivo avaliar o potencial de degradação de MC pela bactéria Sphingosinicella microcystinivorans B9, diferentes cepas de leveduras e bactérias probióticas. O teste foi efetuado com extrato de MC e diferentes quantidades de biovolume e densidade celular dos microrganismos. Os tratamentos foram mantidos a 27ºC com rotação de 100 rpm e as amostras para análise de MC e contagem dos microrganismos foram retiradas após 0 e 96 horas de contato. A bactéria B9 apresentou maior degradação de MC, chegando a 98% após 96 horas.
To minimize problems related to the occurrence of cyanobacteria in water for human consumption there is need to investigate alternative treatment techniques with emphasis on biofilms with the potential degradation of microcystins (MC). This study aimed to evaluate the potential degradation of MC by bacteria Sphingosinicella microcystinivorans B9, different strains of yeast and probiotic bacteria. The test was carried out with the extract obtained from strain Microcystis sp. In the tests biomass and cultures of microorganisms were used and the treatments were maintained at 27ºC with 100 rpm. Samples for analysis of MC and for counting the microorganisms were collected at 0 and 96 hours. The bacterium B9 presented the highest potential of degradation of MC reaching 98% after 96 hours.
ABSTRACT
This work investigated the effects of co-occurring aflatoxin B1 (AFB1) and microcystin (MC) in aquaculture, using immunohistochemistry and genotoxicity methods. Tilapia (Oreochromis niloticus) were exposed to AFB1 by intraperitoneal and MC (cell extract of Microcystis aeruginosa) by intraperitoneal and immersion routes. The interaction of MC-AFB1 was evaluated co-exposing the intraperitoneal doses. Blood samples were collected after 8, 24, and 48h to analyze the micronucleus frequency and comet score. The interaction of MC-AFB1 showed a synergic mutagenic response by higher micronucleus frequency of co-exposed group. A slight genotoxic synergism was also observed in the comet score. Immunohistochemistry detected MC in al lthe fish liver tissues exposed to MC by intraperitoneal route, and only the immersed group with the highest dose of MC showed a positive response. Although MC was non-detectable in the edible muscle, the combination of immunohistochemistry with genotoxicity assay was an attractive biomonitoring tool in aquaculture, where the animals were frequently exposed to co-occurring synergic hazards.
ABSTRACT
A deterioração da qualidade de água pela piscicultura associa-se à eutrofização, com florescimento de cianobactérias. Microcystis aeruginosa destaca-se como principal produtora de microcistinas (MCs), grupo de hepatotoxinas com potencial promotor de tumor. No presente trabalho desenvolveu-se método imunoistoquímico para a detecção de MC em tilápias (Oreochromis niloticus) submetidas à injeção intraperitoneal (i.p.) ou imersão em extrato de M. aeruginosa BCCBUSP 262, empregando anticorpo monoclonal anti-MC (M8H5) e sistema polímero-peroxidase. As tilápias (N=42) foram submetidas a sete tratamentos, sendo três grupos inoculados i.p. com 2,0x105, 4,0x105 e 1,0x106 cels.Kg-1 de M. aeruginosa BCCBUSP 262 e quatro submetidos à imersão em diferentes concentrações do extrato da cianobactéria (variando de 1,0x104 a 1,0x105cel.mL-1). Analisando fígado e tecido muscular pelo ensaio imunoistoquímico, não se detectou marcação em tecido muscular. Todos os animais inoculados i.p. apresentaram marcação positiva para MC no fígado, mas em teste de imersão, apenas os expostos a maior dose (1,0x105 cels.mL- 1) apresentaram marcação positiva. Embora MC não seja detectada em tecido muscular, assim como no fígado de animais imersos em extrato de M. aeruginosa CCBUSP 262 em concentrações menores que 1,0x105 cels.mL-1, os resultados constituíram-se base para o desenvolvimento metodológico objetivando a aplicação da imunoistoquímica no diagnóstico rápido no controle de qualidade de pescados.
The deterioration of the water quality due to aquaculture is associated with eutrophication, with bloom of cyanobacteria. Microcystis aeruginosa is distinguished as main producer of microcystins (MCs), group of hepatotoxins with tumor promoter potential. In the present work immunohistochemical method for detection of MC in tilápia (Oreochromis niloticus), fish submitted to intraperitoneal injection (i.p.) or immersion in extract of M. aeruginosa BCCBUSP 262 was developed, using monoclonal antibody anti- MC (M8H5) and polymer peroxidase system. The tilápias (N=42) had been submitted to the seven treatments, three groups inoculated i.p. with 2.0x105, 4.0x105 and 1.0x106 cells. Kg-1 of M. aeruginosa BCCBUSP 262 and four groups exposed to the immersion in different extract concentrations of cyanobacterium. Analyzing liver and muscular tissue for immunohistochemical assay, muscular tissue was not stained. All the animals inoculated i.p. presented positive marking for MC in the liver, but in immersion test, only the ones exposed in the highest dose (1,0x105 cels.mL-1) presented positive marking. Although MC was not detected in muscular tissue, as well as in the liver of animals immersed in extract of M. aeruginosa BCCBUSP 262 in concentrations less than 1.0x105 cels.mL-1, the results would constitute in the base for the methodological development aiming the application of the immunohistochemistry in the rapid diagnosis in quality control of fish.