Outbreaks of an ulcerative and haemorrhagic disease in Arctic char Salvelinus alpinus caused by Aeromonas salmonicida subsp. smithia

Arctic char Salvelinus alpinus farmed in different places in Austria and free of the viral diseases viral haemorrhagic septcaemia (VHS), infectious haematopoietic necrosis (IHN) and infectious pancreatic necrosis (IPN) experienced disease and mortality. Diseased fish showed skin ulceration and pathological signs of sepsis. Aeromonas sp. was isolated as pure culture from the kidney of freshly euthanized diseased fish. Three independent isolates from outbreaks that occurred on 2 of the affected farms were analyzed phylogenetically by DNA sequence analysis of the rrs and gyrB genes and phenotypically with biochemical reactions. All 3 isolates were identified as Aeromonas salmonicida subsp. smithia. Analysis of virulence genes in these isolates revealed the presence of a Type III secretion system as well as several related virulence effector genes including aexT, encoding the Aeromonas exotoxin AexT, aopP and aopH. These genes are characteristic for virulent strains of typical and atypical subspecies of A. salmonicida.

Role of Aeromonas hydrophila in bacterial septicemia of cultured carps in Khouzestan Province and investigation on protectivity of bacterin (s) prepared from acute isolate(s) of it

Motile Aeromonas are the most common bacteria of freshwater in the world that cause disease in fish and other cold-blooded and warm-blooded hosts. Among this group of bacteria, Aeromonas hydrophila is important in causing complications such as fin rot, skin ulcers and lethal hemorrhagic septicemia in fish. Several virulence factors involved in the pathogenesis of Aeromonas hydrophila, including extracellular enzymes (protease, lipase, elastase, gelatinase and nuclease) and toxins. From the exotoxins, hemolysin, aerolysin and cytolytic enterotoxin play an important role in pathogenesis. Detection of virulence markers by PCR as a key component of determining the pathogenesis of the bacteria and using indigenous vaccines for better immunization against this disease is important. In this study, a total of 200 fanned carps (126 common carp. 39 silver carp and 35 of grass carp) with symptoms suspected aeromonas septicemia were isolated from Khouzestan province farms. 125 bacteria belong to Aeromonas genus detected by biochemical and PCR methods. 31 of all isolates recognized as Aeromonas hydrophila with biochemical methods, I6srRNA detection and Lipase genes. Results showed that the role of Aeromonas sp. and Aeromonas hydrophila in fish with disease symptoms were 62.5% and 15.5% respectively. By using specific primers, three virulence genes including hemolysin, aerolysine and cytolytic enterotoxin were detected in these confirmed isolates, that 18 isolates (58/06%) hemolysin positive (hlyA +), 16 isolates (51/61%) aerolysine positive (aerA+) and 23 isolates (74/19%) for cytolytic enterotoxin gene (act+) were positive. The result of present study showed that most of the confirmed isolates genotype was hlyA+ act- with frequency equal to 51/61%. For investigating the protection effect of acut strain of bacteria, UV inactivated bacterin was used.

Deterioração bacteriológica do jaraqui Semaprochilodus spp. capturado no estado do Amazonas e conservado em gelo

Neste trabalho foram analisados 73 espécimes de jaraqui Semaprochilodus spp. conservados em caixas de poliestireno expandido entre camadas de gelo. Foram realizadas as seguintes análises: avaliação sensorial pela tabela de Torry modificada e pelo índice de qualidade por deméritos; determinação do pH e das bases voláteis totais (N-BVT); contagem total dos microrganismos aeróbios psicrófilos a 20 ºC por 4 dias, psicrotróficos a 7 ºC por 10 dias, dos mesófilos a 37 ºC por 2 dias; contagem, isolamento e identificação das bactérias Aeromonas sp. Bacillus sp. e Pseudomonas sp. a 20 ºC por 24 horas e de Plesiomonas sp. a 37 ºC por 2 dias. O jaraqui se manteve em condições de consumo, pela avaliação sensorial, por 18 e 21 dias. O pH e as bases voláteis totais não foram bons indicadores de qualidade; as contagens totais de psicrófilos, psicrotróficos e mesófilos não apresentaram diferença significativa e as bactérias não apresentaram comportamento deteriorador pela ausência da produção de H2S.

Antibody Based Diagnostic for Detection Of Vibrios and their Biological Control using Antagonistic Bacteria in Macrobrachium Rosenbergii Larval Rearing Systems

The main objective of the work undertaken here was to develop an appropriate microbial technology to protect the larvae of M.rosenbergii in hatchery from vibriosis. This technology precisely is consisted of a rapid detection system of vibrios and effective antagonistic probiotics for the management of vibrios. The present work was undertaken with the realizations that to stabilize the production process of commercial hatcheries an appropriate, comprehensive and fool proof technology is required primarily for the rapid detection of Vibrio and subsequently for its management. Nine species of Vibrio have been found to be associated with larvae of M. rosenbergii in hatchery. Haemolytic assay of the Vibrio and Aeromonas on prawn blood agar showed that all isolates of V. alginolyticus and Aeromonas sp., from moribund, necrotized larve were haemolytic and the isolates of V.cholerae, V.splendidus II, V.proteolyticus and V.fluvialis from the larvae obtained from apparently healthy larval rearing systems were non-haemolytic. Hydrolytic enzymes such as lipase, chitinase and gelatinase were widespread amongst the Vibrio and Aeromonas isolates. Dominance of V.alginolyticus among the isolates from necrotic larvae and the failure in isolating them from rearing water strongly suggest that they infect larvae and multiply in the larval body and cause mortality in the hatchery. The observation suggested that the isolate V. alginolyticus was a pathogen to the larvae of M.rosenbergii. To sum up, through this work, nine species of Vibrio and genus Aeromonas associated with M.rosenbergii larval rearing systems could be isolated and segregated based on the haemolytic activity and the antibodies (PA bs) for use in diagnosis or epidemiological studies could be produced, based on a virulent culture of V.alginolyticus. This could possibly replace the conventional biochemical tests for identification. As prophylaxis to vibriosis, four isolates of Micrococcus spp. and an isolate of Pseudomonas sp. could be obtained which could possibly be used as antagonistic probiotics in the larval rearing system of M.rosenbergii.

A marine bacterium, Micrococcus MCCB 104, antagonistic to vibrios in prawn larval rearing systems

marine bacterium, Micrococcus MCCB 104, isolated from hatchery water, demonstrated extracellular antagonistic properties against Vibrio alginolyticus, V. parahaemolyticus, V. vulnificus, V. fluviallis, V. nereis, V. proteolyticus, V. mediterranei, V. cholerae and Aeromonas sp., bacteria associated with Macrobrachium rosenbergii larval rearing systems. The isolate inhibited the growth of V. alginolyticus during co-culture. The antagonistic component of the extracellular product was heat-stable and insensitive to proteases, lipase, catalase and α-amylase. Micrococcus MCCB 104 was demonstrated to be non-pathogenic to M. rosenbergii larvae

Culex pipiens development is greatly influenced by native bacteria and exogenous yeast

Culex pipiens is the most cosmopolitan mosquito of the Pipiens Assemblage. By studying the nature of interactions between this species and microorganisms common to its breeding environment we can unravel important pitfalls encountered during development. We tested the survival rate of larval stages, pupae and adults of a Cx. pipiens colony exposed to a variety of microorganisms in laboratory conditions and assessed the transmission to offspring (F1) by those organisms that secured development up to adulthood. Three complementary experiments were designed to: 1) explore the nutritional value of yeasts and other microorganisms during Cx. pipiens development; 2) elucidate the transstadial transmission of yeast to the host offspring; and 3) to examine the relevance of all these microorganisms in female choice for oviposition-substratum. The yeast Saccharomyces cerevisiae proved to be the most nutritional diet, but despite showing the highest survival rates, vertical transmission to F1 was never confirmed. In addition, during the oviposition trials, none of the gravid females was attracted to the yeast substratum. Notably, the two native bacterial strains, Klebsiella sp. and Aeromonas sp., were the preferred oviposition media, the same two bacteria that managed to feed neonates until molting into 2nd instar larvae. Our results not only suggest that Klebsiella sp. or Aeromonas sp. serve as attractants for oviposition habitat selection, but also nurture the most fragile instar, L1, to assure molting into a more resilient stage, L2, while yeast proves to be the most supportive diet for completing development. These experiments unearthed survival traits that might be considered in the future development of strategies of Cx. pipiens control. These studies can be extended to other members of the Pipiens Assemblage

Microbiological quality of salmon (Salmo salar) sold in cities of the state of São Paulo, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Características microbiológicas de salmão (Salmo salar) comercializado em algumas cidades da região nordeste do estado de São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Infecções parasitárias e microbianas na produção do pacu Piaractus mesopotamicus e do híbrido patinga procedentes da região noroeste do Estado de São Paulo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ecotoxicidade, segurança clínica e eficácia de fármacos em jovens de pacu (Piaractus mesopotamicus)

Pós-graduação em Aquicultura - FCAV

Effectiveness of therapeutic agents in disease treatment in Piaractus mesopotamicus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

De um subproduto industrial para um produto destinado à alimentação animal: produção de biomassa bacteriana

A grande diversidade das atividades industriais ocasiona durante o processo produtivo a geração de resíduos sólidos, líquidos e gasosos, que podem poluir e/ou contaminar o solo, a água e o ar. Embora existam regulamentos para o descarte desses resíduos, a inobservância às regras, a ineficiência dos tratamentos despoluentes e a inoperância de órgãos fiscalizadores permitem que, ainda hoje, sejam lançados ao ambiente grandes cargas de poluentes. Isso se aplica às indústrias de alimentos de origem animal. Felizmente, a visão dos empresários do setor vem mudando, na medida em que a imagem de uma empresa que conta com produtos e processos ambientalmente responsáveis representa parte das estratégias competitivas atuais. O objetivo deste trabalho foi demonstrar como um efluente industrial pode ser transformado em matéria prima para a obtenção de um produto com valor comercial, utilizando tecnologias reconhecidas pela indústria e a atividade microbiana. O efluente foi obtido em indústria de abate e processamento de tilápias, apresentado (valores médios) pH = 9,4, DQO = 1.127 mg/L, óleos e graxas = 1.166 mg/L, nitrogênio total = 813 mg/L, coliformes a 30–35o C = 1,0x105 NMP/mL, coliformes a 45o C = 0,41 NMP/mL, bolores e leveduras = 4,6x103 UFC/mL e, ocasionalmente, contendo Salmonella sp e Aeromonas sp. Os tratamentos físicos aplicados ao efluente incluíram gradeamento, filtração (50µm) e pasteurização (65o C/30 min). O cultivo de Rubrivivax gelatinosus foi realizado sob anaerobiose em reatores de vidro durante 7 dias, em temperatura ambiente (30±5o C) e 2.000±500 lux. A recuperação da biomassa foi feita por filtração tangencial (0,2 µm; 1,5 bar), centrifugação (3.400 g/30 min; 5o C) e liofilização (-40o C) e a pulverização foi realizada manualmente. A produção de massa celular atingiu 0,18 g/L, com produtividade de 0,0634 g/L.dia. O processo promoveu redução de 52% na DQO, 48% em óleos e graxas e 22% no nitrogênio total, gerando um resíduo com pH 7,9, livre de bactérias patogênicas e, portanto, apto ao descarte. O produto obtido apresentou cor vermelho escuro (L = 22,42; C = 14,22; h = 25,48), 4,55% de umidade, 57,39% de proteína, 11,08% de extrato etéreo, 4,05% de matéria mineral, 3,03 mg/g de oxicarotenóides, 20,27 NMP/g de coliformes a 30–35o C, <1,0 NMP/g de coliformes a 45o C, 1,2x103 UFC/g de bolores e leveduras e ausência de microrganismos patogênicos. Essas características apontam para um potencial pigmentante e nutricional positivo no produto, que pode encontrar aplicação na produção animal, com segurança microbiológica. Dessa forma, fecha-se um ciclo autosustentável que pode ser adotado na própria fonte geradora do resíduo industrial, permitindo a reversão de gastos com tratamento despoluente em receitas advindas da comercialização de um novo produto.

Protocolo para el uso y aplicación racional de productos químicos, microbiológicos y antibióticos en la producción de camarón marino de cultivo en El Salvador

Este protocolo tiene por objeto establecer los requisitos y medidas sanitarias de prevención y control de la dispersión de enfermedades de alto impacto y para el uso adecuado y racional de fármacos de uso específico en la acuicultura, específicamente para la producción camaronera y de los laboratorios de producción de nauplios y post larvas del país. Este documento es resultado del proyecto de investigación denominado Muestreo y diagnóstico de la calidad microbiológica del agua durante un ciclo de cultivo de camarón marino en estanques de cooperativas de San Hilario, Bahía de Jiquilisco, Usulután, ejecutado por la Escuela Especializada en Ingeniería ITCA-FEPADE, Centro Regional La Unión. En este proyecto se determinó cualitativa y cuantitativamente la presencia de coliformes totales, coliformes fecales (Es-cherichia coli) y bacterias Heterótrofas que constituyen los bioindicadores estándar de contaminación orgánica del agua. Se determinó además la presencia de Vibrio sp., Pseudomonas sp., y Aeromonas sp. en el área de estudio identificados como los principales agentes causantes de enfermedades infecciosas en el camarón marino. El muestreo ayudó a conocer la influencia de parámetros físico-químicos en la presencia de los microorganismos estudiados y servirá de marco para elaborar una propuesta para un programa de monitoreo de la acuicultura en todas sus etapas del encadenamiento productivo. Se realizó un taller con la Cooperativa Senderos de Paz sobre los resultados de TCBS de la calidad del agua en el canal reservorio y el estanque número 3 de dicha cooperativa.

Common fish diseases and parasites affecting wild and farmed tilapia and catfish in central and western Uganda

Intensification of aquaculture production in Uganda is likely to result into disease out-breaks leading to economic losses to commercial fish farms and associated natural aquatic ecosystems. This survey assessed health profiles of selected commercial fish farms and adjacent natural aquatic ecosystemsto identify fish diseases and parasites affecting Nile tilapia (Oreochromis niloticus) and African catfish (Clarias gariepinus) in aquaculture systems in Uganda. Fish farms encounter disease out-breaks that cause low survival rates (0 - 30%), especially catfish hatcheries. Health management issues are not well understood by fish farmers, with some unable to detect diseased fish. Current control strategies to control aquatic pathogens include use of chemotherapeutants and antibiotics. Bacterial pathogens isolated included Flavobacterium columnare, Aeromonas sp., Edwardsiella sp., Psuedomonus sp., Steptococcus sp., Staphylococcus sp., Proteus sp., and Vibrio sp. A high occurrence of Flavobacterium columnare exists in both asymptomatic and symptomatic fish was observed. Parasites included protozoans (Ichthyopthirius multiphilis, Trichodina sp. and Icthyobodo sp.) and trematodes (Cleidodiscus sp. and Gyrodactylus sp.). Diagnosis and control of diseases and parasites in aquaculture production systems requires adoption of a regional comprehensive biosecurity strategy: the East African (EAC) region unto which this study directly contributes.

Characterization and virulence potential of phenotypically diverse Aeromonas veronii isolates recovered from moribund freshwater ornamental fishes of Kerala, India

In the present study, we investigated the involvement of Aeromonas spp. in eliciting disease outbreaks in freshwater ornamental fishes across the state of Kerala, India. We investigated three incidences of disease, in which the moribund fishes exhibited clinical signs such as haemorrhagic septicemia (in gouramy, Trichogaster sp.), dropsy (in Oscar, Astronotus ocellatus) and tail rot/fin rot (in gold fish, Carassius carassius). Pure cultures (n = 20 from each fish; 60 in total) of Aeromonas spp. were recovered from the abdominal fluid as well as from internal organs of affected fishes, although they could not be identified to species level because of the variations in their phenotypic characters. The molecular fingerprinting of the isolates using Enterobacterial Repetitive Intergenic Consensus PCR proved the genetic diversity of the isolates from the three sites. The phylogenetic trees constructed using concatenated sequences (using 16S rRNA, gyrA, gyrB and rpoD genes) indicated that they were related to Aeromonas veronii. They exhibited marked cytotoxic and haemolytic activity, which were responsible for the pathogenic potential of the isolates. The isolates possessed multiple virulence genes such as enterotoxins (act and alt), haemolytic toxins (aerA and hlyA), genes involved in type III secretion system (ascV, aexT and ascF–ascG), glycerophospholipid-cholesterol acyltransferase (gcat) and a type IV pilus (tapA) gene, as determined by PCR. Virulence of representative isolates to goldfish was also tested, and we found LD50 values of 104.07–105.35 cfu/fish. Furthermore, the organisms could be recovered as pure cultures from the lesions as well as from the internal organs.