Resumo: O objetivo deste trabalho foi avaliar o efeito do uso de substrato artificial para o crescimento de biofilme perifítico sobre a composição microbiológica do biofilme, a qualidade da água e o desempenho zootécnico de tilápia-do-nilo (Oreochromis niloticus), em tanques de cultivo. O experimento consistiu de dois tratamentos: presença e ausência de substrato artificial para o crescimento de perifíton, com cinco repetições. Os parâmetros de qualidade de água foram avaliados durante todo o período experimental. A cada duas semanas, foram coletadas amostras do perifíton para análise da sua composição microbiológica e realizadas medidas biométricas dos peixes. A quantidade de bactérias heterotróficas no biofilme aumentou durante o período experimental, mas a de Aeromonas reduziu com o uso do substrato artificial. O nitrogênio amoniacal total dos tanques com perifíton reduziu 30% em relação ao do controle. As tilápias criadas com o biofilme apresentaram peso final 2,4 vezes maior que às submetidas ao controle, além de aumento de 59,19% na sua produtividade. O uso de substrato artificial para a formação de biofilme perifítico em cultivo de tilápia em tanques favorece a manutenção da qualidade da água de cultivo, a proteção dos peixes contra patógenos e a melhora do desempenho zootécnico dos peixes.

Abstract: The objective of this work was to evaluate the effect of the use of an artificial substrate for periphytic biofilm growth on the microbiological composition of the biofilm, water quality, and zootechnical performance of Nile tilapia (Oreochromis niloticus) in rearing tanks. The experiment consisted of two treatments: presence and absence of artificial substrate for the growth of periphyton, with five replicates. Water quality parameters were evaluated throughout the entire experimental period. Every two weeks, samples of the periphyton were collected for the analysis of its microbiological composition and biometric measurements of the fish were performed. The number of heterotrophic bacteria in the biofilm increased during the experimental period, but that of Aeromonas decreased with the use of the artificial substrate. The total ammonia nitrogen in the tanks with periphyton reduced in 30% compared with that of the control. Tilapia reared with the biofilm showed a final weight 2.4 times greater than that of those subjected to the control, as well as an increase of 59.19% in their productivity. The use of an artificial substrate for periphytic biofilm in Nile tilapia rearing tanks favors the maintenance of the quality of the culture water, the protection of the fish against pathogens, and the improvement of the zootechnical performance of the fish.

ABSTRACT. This study aims at investigating to follow the formation and development of biofloc aggregates in a system with the introduction of an in vitro selected bacterial consortium (Bacillus thuringiensis, Bacillus sp., Staphylococcus cohnii) in order to induce fast formation of biofloc and to compare it to the development of spontaneous formation biofloc. Two experimental groups were evaluated for biofloc formation, SFT and IFT. The first refers to spontaneous (conventional) formation of the flocs and the second to induced formation (IFT), achieved through the consortium of potentially inducing bacteria. Both treatments presented a constant increase of bioflocs, however, in the IFT treatment, the microbial aggregates were larger and more uniform. By the end of the experiment, we verified that the aggregates formed in the IFT showed higher volume and lower sedimentation rate in comparison to the spontaneously formed ones. The results show that domestication in microbial communities is efficient as related to bioflocs, reducing instability during its formation and development.

ABSTRACT Microbiota, the microorganisms that colonize living organisms. Oysters rely exclusively on an innate immune system. This study characterized the cultivable gastrointestinal tract microbiota (GTM) of the oyster Crassostrea gasar and evaluated their influence on immune responses. Antibiotics were used to reduce or alter the microbiota during in vitro and in vivo assays. Haemocyte viability, concentration, and phagocytic capacity, as well as the production of reactive oxygen species (ROS), were evaluated in antibiotic-treated and control oysters. Microbiological analysis of the gastrointestinal tract was also performed; bacteria were selected using culture media and were identified by 16S ribosomal DNA. The results showed that the antibiotics eliminated bacteria in vitro but increased their concentrations in vivo. The GTM was composed of 26% amylolytic bacteria, 21% lipolytic bacteria, 18% proteolytic bacteria, 18% cellulolytic bacteria and 17% lactic acid bacteria; there were no differences in the amounts of these bacterial types between the control and treated oysters. Three major bacterial phyla, Proteobacteria, Firmicutes and Actinobacteria, and seven genera, Labrenzia, Pseudomonas, Halomonas, Shewanella, Vibrio, Bacillus, and Micrococcus, were detected. The concentration, viability, and phagocytic capacity of hemocytes and the production of ROS were not significantly altered by antibiotic treatment. In conclusion, the antibiotics altered the number of heterotrophic bacteria without changing hemocyte function, suggesting that this assay could be useful for verifying the influence of microbiota in host-parasite interactions.

RESUMO A geração de efluentes e o custo com a alimentação do camarão são os principais desafios enfrentados pela carcinicultura. Uma solução conjunta para ambos os problemas é a produção de bioflocos em um sistema de lodo ativado para tratamento de efluentes. Neste trabalho, o afluente ao sistema estudado era composto pelas águas residuais do cultivo de camarão suplementadas com ureia e melaço de cana-de-açúcar. Os resultados mostraram que a remoção média de demanda química de oxigênio foi de 71% e a produção média de bioflocos no reator foi de aproximadamente 1,5 g.L-1. A adição de melaço ao afluente contribuiu para o aumento da quantidade e da diversidade de microrganismos benéficos para a produção de camarão. Houve remoção de amônia e nitrificação confirmada pelo balanço de massa. Portanto, a utilização do sistema de lodos ativados é uma alternativa viável e ambientalmente adequada para produzir bioflocos e tratar efluentes de cultivo de camarão.

ABSTRACT The release of wastewater and the shrimp feed cost are the main challenges faced by the shrimp farming industry. An alternative solution to both problems is biofloc production in a unit external to the farm, in an activated sludge system for effluent treatment. The treatment system’s influent was composed of the shrimp farm wastewater supplemented with urea and sugarcane molasses. The results show that the average removal of chemical oxygen demand was 71% and the average biofloc production in the reactor was approximately 1.5g.L-1. Adding molasses to the influent contributed to the increase in the quantity and diversity of existing microorganisms that are beneficial to cultured shrimp. The mass balance of nitrogen compounds confirmed that nitrification occurred in the system. Therefore, the use of the activated sludge system is a viable and environmentally suitable alternative to produce bioflocs and shrimp farming effluent treatment.

RESUMO A presente pesquisa isolou e identificou os principais grupos bacterianos presentes no cultivo de juvenis de tilápia do Nilo na presença de bioflocos e/ou perifíton. Verificou-se a produção de exoenzimas como indicadoras de potenciais fatores de virulência e a capacidade de formação de biofilme. As amostras de água foram retiradas do cultivo das tilápias na presença de bioflocos (T1), cultivo na presença de bioflocos e perifíton (T2), cultivo tradicional (T3) e cultivo na presença de perifíton (T4). Para o crescimento e seleção dos grupos bacterianos foi utilizada a técnica da semeadura em profundidade com os seguintes meios de cultura: Ágar para Contagem em Placas (PCA - DIFCO), Aero Pseudo Seletive Ágar (GSP - Himedia) e Ágar Nutriente (AN - Merck). Foram isoladas 46 cepas, distribuídas da seguinte forma: T1 (n = 12); T2 (n = 10); T3 (n = 14) e T4 (n = 10). Entre os isolados, os gêneros mais frequentes foram: Pseudomonas spp., Aeromonas spp., Staphylococcus spp., Bacillus spp., Mycobacterium spp., Micrococcus spp., e Corybacterium spp. Os isolados bacterianos dos tratamentos T1 e T3 expressaram cinco perfis de virulência enquanto aqueles isolados dos tratamentos T2 e T4 apresentaram dois e três perfis de virulência, respectivamente. Os tratamentos com presença de bioflocos e perifíton (T2) ou somente perifíton (T4) apresentaram bactérias com menor potencial patogênico. Em relação ao crescimento dos peixes, o T1 e T4 possibilitaram maior ganho em peso final.

ABSTRACT The following research isolated and identified the main bacterial groups present in the culture of juvenile Nile tilapia in the presence of bioflocs and/or periphyton. The strains were also tested for the production of exoenzymes, indicative of potential virulence factors, and ability to form biofilm. The water samples were taken from tilapia cultured in the presence of bioflocs (T1), in the presence of bioflocs and periphyton (T2), from traditional culture (T3) and from culture in the presence of periphyton (T4). In the growth and selection of the bacterial groups, pour plate method was used, along with the following media: Plate Count Agar (PCA - DIFCO), Aero Pseudo Selective Agar (GSP - Himedia) and Nutrient Agar (AN - Merck). 46 strains were isolated in the following distribution: T1 (n = 12); T2 (n = 10); T3 (n = 14) and T4 (n = 10). Among the isolates, the most frequent genera were: Pseudomonas spp., Aeromonas spp., Staphylococcus spp., Bacillus spp., Mycobacterium spp., Micrococcus spp., and Corybacterium spp. Bacterial isolates in treatments T1 and T3 tested positive for five virulence profiles each, while those isolated from T2 and T4 for two and three virulence profiles, respectively. Treatments in bioflocs and periphyton (T2) or only periphyton (T4) yielded bacteria of less pathogenic potentials. In relation to the fish growth, T1 and T4 resulted in a higher final weight.

RESUMO: Nos últimos anos o surgimento de patógenos resistentes tem dificultado o tratamento de infecções bacterianas, tanto na produção animal como na área médica, em decorrência do desenvolvimento de mecanismos de resistência pelos micro-organismos. O objetivo deste estudo foi traçar o perfil de resistência antimicrobiana em cepas de Salmonella spp. isoladas de moluscos bivalves (ostras e sururu) e da agua de áreas estuarinas em duas regiões da Bahia, Brasil. Um total de 27 cepas previamente isoladas foram testadas, sendo 12 provenientes de moluscos bivalves e 15 de amostras de água. Para o teste de suscetibilidade foram utilizados oito agentes antimicrobianos (classes fenicóis, betalactâmicos, tetraciclina, quinolonas e fluoroquinolonas), Concentração Inibitória Mínima (CIM) e a produção de Betalactamases de Espectro Ampliado (ESBL). Os isolados apresentaram alta suscetibilidade as classes de antimicrobianos testadas, com resistência apenas ao ácido nalidíxico (27%), ampicilina (25%) e tetraciclina (25%). A resistência bacteriana era de origem cromossômica e o índice de multirresistência (MAR) entre os isolados de molusco (sururu in natura) foi de 0,25. A CIM encontrada foi de 100 µg/mL, 500 µg/mL e 350 µg/mL para o ácido nalidixíco, ampicilina e tetraciclina, respectivamente. Nenhum dos isolados apresentou produção de ESBL. A presença de Salmonella spp. multirresistente e com elevada CIM está sendo veiculada em áreas de extração de moluscos bivalves no Estado da Bahia, Brasil.

ABSTRACT: In recent years, the emergence of resistant pathogens has complicated the treatment of bacterial infections in livestock production as well as in the medical field, due to the development of resistance mechanisms by microorganisms. The objective of this study was to delineate the antimicrobial resistance profile of Salmonella spp. strains isolated from bivalve mollusks (oysters and mussels) and from estuarine environment water of two regions of Bahia, Brazil. Twenty-seven strains, 12 isolated from bivalve mollusks and 15 from estuarine water, were tested. Eight antimicrobial agents (phenicol, beta-lactams, tetracyclines, quinolones and fluoroquinolones classes) were used for a susceptibility test, Minimum Inhibitory Concentration (MIC) and extended-spectrum beta-lactamases (ESBLs) production. Isolates showed high susceptibility to the classes of antimicrobial agents tested, with resistance only to nalidixic acid (27%), ampicillin (25%) and tetracycline (25%). Bacterial resistance was of chromosomal origin and the multidrug resistance index (MAR) among isolates of shellfish (mussels in natura) was 0.25. The MIC was found to be 100 µg/mL, 500 µg/mL and 350 µg/mL to nalidixic acid, ampicillin and tetracycline, respectively. None of the isolates presented ESBLs production. The presence of multidrug-resistant and high MIC Salmonella spp. is being conveyed in extraction areas of bivalve mollusks in the State of Bahia, Brazil.

O objetivo deste estudo foi detectar a presença potencial virulência de Vibrio cholerae isolado de estuários do Nordeste do Brasil. Amostras de água e sedimento foram coletadas e inoculadas sobre meio seletivo para víbrios (TCBS) e colônias com características morfológicas de V. cholerae foram isoladas. A identificação fenotípica seguiu chave dicotômica baseada em caraterísticas bioquímicas. Foram empregadas as técnicas de amplificação da polimerase em cadeia (PCR) utilizando o gene ompW e a de multiplex PCR para detecção de genes de virulência (ctx, tcp, zot e rfbO1). Os resultados da identificação das diferentes abordagens foram comparados. Nove cepas de V. cholerae foram identificadas fenotipicamente e cinco confirmadas através da detecção do gene ompW. A chave dicotômica utilizada foi eficiente para a confirmação da espécie. Os quatro estuários analisados apresentaram estirpes de V. cholerae, e nenhuma das cepas isoladas apresentaram genes de virulência.

The objectives of this study were to detect the presence of Vibrio cholerae in tropical estuaries (Northeastern Brazil) and to search for virulence factors in the environmental isolates. Water and sediment samples were inoculated onto a vibrio-selective medium (TCBS), and colonies with morphological resemblance to V. cholerae were isolated. The cultures were identified phenotypically using a dichotomous key based on biochemical characteristics. The total DNA extracted was amplified by PCR to detect ompW and by multiplex PCR to detect the virulence genes ctx, tcp, zot and rfbO1. The results of the phenotypic and genotypic identification were compared. Nine strains of V. cholerae were identified phenotypically, five of which were confirmed by detection of the species-specific gene ompW. The dichotomous key was efficient at differentiating environmental strains of V. cholerae. Strains of V. cholerae were found in all four estuaries, but none possessed virulence genes.

Relato de surto familiar de botulismo por intoxicação alimentar, envolvendo um óbito, onde foram encontradas lacunas no preenchimento do prontuário. O objetivo foi descrever a patologia chamando a atenção dos profissionais de saúde para o fornecimento adequado de informações relevantes para a investigação epidemiológica de doenças de notificação compulsória.

Report of a family outbreak of botulism food poisoning involving a death, where gaps in the completion of medical records were identified. The study aimed to describe the pathology and emphasize to health professionals the need to provide adequate information relevant to epidemiological investigation of compulsory notification diseases.

The aims of this study were to count and identify sucrose positive and negative vibrios isolated from cultivated Crassostrea rhizophorae oysters during their growing cycle. Every month for 12 months, 10 to 18 oysters were collected for study. Collections occurred at the Center for Studies of Coastal Aquaculture (CSCA), which is associated with the Institute of Marine Science, Labomar, located in Euzebio, Ceará, Brazil. Approximately 150 oysters and their intervalvular liquor were studied. Vibrio Standard Plates Counts (SPC) from oyster meat and their intervalvular liquor varied from 25 to 59,000,000 CFU/g. For most of the 12 months of the oysters' life, it was possible to identify Vibrio parahaemolyticus. Vibrio carchariae was identified in four collections. Among other isolated species, the most important, considering public health risks, was V. vulnificus, although only one strain was confirmed. We concluded that retail purchased oysters should never be eaten raw or undercooked because many species of the genus Vibrio are known to be pathogenic to humans and live naturally on and in shellfish throughout their life cycle.

O chá de folhas de goiaba Psidium guajava Linnaeus é comumente usado como remédio nas gastrenterites e diarréias infantis por aqueles que não têm acesso a antibióticos. Esta pesquisa estudou o efeito antibacteriano sobre bactérias causadoras de diarréias, do óleo essencial e do extrato de folhas de goiabeira usando como diluente: metanol, hexano e acetato de etila. Os extratos foram testados sobre Staphylococcus aureus, Salmonella spp. e Escherichia coli. As bactérias testadas foram isoladas de camarão sete-barbas Xiphopenaeus kroyeri (Heller) usando-se como controle cepas padrão, de cada espécie. Das bactérias testadas, o melhor efeito inibitório foi observado sobre Staphylococcus aureus. O extrato de metanol apresentou maior inibição bacteriana. O óleo essencial mostrou ação inibitória contra S. aureus e Salmonella spp. Foi demonstrada alguma resistência das cepas isoladas de camarão aos antibióticos comerciais testados. Estes dados confirmam que remédios à base de folhas de goiabeira podem ser utilizados em casos de diarréia provocada por essas bactérias, quando o uso de antibiótico for restrito. Conclui-se que extratos de folhas de goiaba e o seu óleo essencial são muito ativos contra S. aureus, tornando-os importantes fontes em potencial de novos compostos antimicrobianos.

Guava leaf tea of Psidium guajava Linnaeus is commonly used as a medicine against gastroenteritis and child diarrhea by those who cannot afford or do not have access to antibiotics. This study screened the antimicrobial effect of essential oils and methanol, hexane, ethyl acetate extracts from guava leaves. The extracts were tested against diarrhea-causing bacteria: Staphylococcus aureus, Salmonella spp. and Escherichia coli. Strains that were screened included isolates from seabob shrimp, Xiphopenaeus kroyeri (Heller) and laboratory-type strains. Of the bacteria tested, Staphylococcus aureus strains were most inhibited by the extracts. The methanol extract showed greatest bacterial inhibition. No statistically significant differences were observed between the tested extract concentrations and their effect. The essential oil extract showed inhibitory activity against S. aureus and Salmonella spp. The strains isolated from the shrimp showed some resistance to commercially available antibiotics. These data support the use of guava leaf-made medicines in diarrhea cases where access to commercial antibiotics is restricted. In conclusion, guava leaf extracts and essential oil are very active against S. aureus, thus making up important potential sources of new antimicrobial compounds.

As ostras são alimentos marinhos freqüentemente ingeridos crus ou parcialmente cozidos. Por esta razão, o risco para a saúde dos consumidores desses produtos é muito elevado, principalmente, quando são de regiões tropicais. Foi estudada a presença de Vibrio cholerae e Vibrio parahaemolyticus em ostras de um estuário na região Nordeste do Brasil. Trezentas ostras foram analisadas, em um período de 8 meses. A salinidade da água, no local de coleta, variou de 3 a 27‰. V. cholerae foi o vibrio mais freqüentemente detectado (33,3% das amostras). Dos 22 isolados, 20 foram identificados como V. cholerae não-O1/não-O139, duas delas apresentando forma rugosa sendo a maioria das demais pertencente ao tipo fermentativo Heiberg II. V. parahaemolyticus foi isolado em apenas umas das coletas. Foram, também, identificadas nas amostras isolados de Providencia spp., Klebsiella spp., Proteus spp. e Morganella morganii.

Oysters are edible organisms that are often ingested partially cooked or even raw, presenting therefore a very high risk to the consumers' health, especially in tropical regions. The presence of Vibrio cholerae and Vibrio parahaemolyticus in oysters sampled at an estuary in the Brazilian northeastern region was studied, with 300 oysters tested through an 8-months period. The salinity of the water at the sampling point varied between 3% and 27‰. V. cholerae was the most frequently detected species (33.3% of the samples), and of the 22 V. cholerae isolates, 20 were identified as non-O1/non-O139, with two of the colonies presenting a rough surface and most of remaining ones belonging to the Heiberg II fermentation group. V. parahaemolyticus was isolated from just one of the samples. Other bacteria such as Providencia spp., Klebsiella spp. and Morganella morganii were also isolated.

Foram coletadas doze amostras de camarão e peixes nas imediações do interceptor oceânico, em Fortaleza e igual número na Feira de pescado do Mucuripe, Fortaleza, para isolamento de E. coli e Staphylococcus aureus, respectivamente. Extratos aquosos, alcoólicos e cetônicos de broto de goiabeira e de folha de mamão foram testados frente às bactérias para se verificar suas ações antibióticas. As cepas de E. coli utilizadas nos ensaios foram as classificadas como LT positivas. Os extratos de folhas de mamão (Carica papaya Linn) não revelaram quaisquer atividades antibióticas enquanto que os preparados com broto de goiabeira (Psidium guajava Linn) apresentaram halos sempre >13 mm para as duas espécies, considerados como de inibição pelo método empregado. Os extratos de broto de goiabeira que apresentaram melhores resultados frente às cepas de E. coli ETEC foram os alcoólicos a 50% seguido do cetônico também a 50%. Concluímos que nos tratamentos de diarréias causadas por E. coli ou por toxinas elaboradas por S. aureus o extrato de brotos de goiabeira é uma opção devido a sua pronta ação curativa, seu fácil cultivo nos países tropicais e ao seu baixo valor aquisitivo.

Out of the twenty-four samples of shrimp and fish muscle used for this study, twelve were collected near a large marine sewer for waste disposal, 3 km off the coast of Fortaleza (Brazil) and used for the isolation of E. coli. Other twelve were collected at the Mucuripe fresh fish market (Fortaleza, Brazil) and used for the isolation of Staphylococcus aureus. Ethanol, water and acetone-diluted extracts of guava and papaya leaf sprouts were tested on the bacteria in order to verify their microbicidal potential. The E. coli strains used in the trials were rated LT positive. The papaya leaf extracts (Carica papaya Linn) showed no microbicidal activity while the guava sprout extracts (Psidium guajava Linn) displayed halos exceeding 13 mm for both species, an effect considered to be inhibitory by the method employed. Guava sprout extracts by 50% diluted ethanol most effectively inhibited E. coli (EPEC), while those in 50% acetone were less effective. It may be concluded that guava sprout extracts constitute a feasible treatment option for diarrhea caused by E. coli or by S. aureus-produced toxins, due to their quick curative action, easy availability in tropical countries and low cost to the consumer.