Isolation and characterization of pathology in case of massive mortality by Photobacterium damselae subsp. piscicida in Rachycentron canadum / Isolamento e caracterização da patologia em caso de mortalidade maciça por Photobacterium damselae subsp. piscicida em Rachycentron canadum

This study aimed to investigate outbreak with high mortality in cultured juvenile cobiaoccurred in Southeast Brazil in 2011. Fish displayed retarded growth rates, lethargy, fin ulceration, skin depigmentation, corneal opacity, and physical deformities. Internally, livers were increased in volume and pale in different degrees. Firm whitish nodules were disseminated in the liver, kidney and spleen. A moderate number of parasites identified as Neobenedenia melleni were recovered from the body surface. Microscopically, severe hepatic steatosis and extensive granulomatous lesion were identified in all fish sampled. Microbiological analysis of moribund fish revealed the presence in pure culture of a Gram-negative bacterium identified as Photobacterium damselae subsp. piscicida using biochemical and molecular characteristics. Analysis of the partial 16S rRNA sequences confirmed the results demonstrating high identity (98%). The isolates were sensitive to chloramphenicol and enrofloxacin and resistant to ciprofloxacin, florfenicol, doxycycline hydrochloride, norfloxacin, oxytetracycline, and tetracycline. Chronic pasteurellosis was considered as the main problem in the farm, while hepatic steatosis and parasitic infestation may have contributed to the development of the process.

Este estudo objetivou investigar um surto com alta mortalidade em cobia juvenis cultivadas na região Sudeste do Brasil em 2011. Os peixes apresentavam baixa taxa de crescimento, letargia, ulceração nas nadadeiras, despigmentação da pele, opacidade da córnea e deformidades físicas. Internamente o fígado apresentava aumentado e pálido em diferentes graus, com nódulos esbranquiçados e firmes disseminados no fígado, rins e baço. Na superfície corporal dos peixes foram observados moderado número de parasitas identificados como Neobenedenia melleni. Microscopicamente verificou-se esteatose hepática grave e extensa lesão granulomatosa em todos os peixes amostrados. A análise microbiológica dos peixes moribundos revelou a presença, em cultura pura de uma bactéria Gram-negativa identificada como Photobacterium damselae subsp. piscicida usando características bioquímicas e moleculares. A análise das sequências parciais de 16S rRNA confirmou os resultados demonstrando alta identidade (98%). Os isolados foram sensíveis a cloranfenicol e enrofloxacina e resistente a ciprofloxacina, florfenicol, cloridrato de doxiciclina, norfloxacina, oxitetraciclina e tetraciclina. A pasteurelose crônica foi considerada como o principal problema na maricultura, enquanto a esteatose hepática e a infestação parasitária podem ter contribuído para o desenvolvimento do processo.

Characterization and utilization of monoclonal antibodies reactive to Yersinia pseudotuberculosis.

The 3 murine monoclonal antibodies, Yps1, Yps2 and Yps3 reactive to Y. pseudotuberculosis can be stabilized and all were found to be of IgG type. Monoclonal antibody, Yps1, recognized a glycoprotein antigen of the organism with reactivity at the 55-75 kDa region, while Yps2 and Yps3 recognized protein antigens of Y. pseudotuberculosis 65 kDa and 26-28 kDa molecular weight regions, respectively. The specificity of monoclonal antibodies was tested using dot ELISA and Western blotting with whole cell organisms or whole cell sonicated soluble antigens of different Yersinia species, Salmonella typhi, Klebsiella pnemoniae, Streptococcus abortus-equi and Escherichia coli. Monoclonal antibody, Yps1 exhibited cross-reactivity with soluble antigens and whole cell preparations of Y. pestis. Yps2 cross-reacted to soluble antigens of all the tested bacteria. Reactivity of monoclonal antibody, Yps3 was restricted to Y. pseudotuberculosis and Y. pestis with soluble antigen preparations. No reaction was observed with Yps2 and Yps3 to whole cell organism preparations from tested bacteria including Y. pseudotuberculosis. The co-agglutination reagent prepared by sensitizing staphylococcal cells with Yps1 monoclonal antibody produced a positive agglutination with all the 4 Y. pseudotuberculosis isolates and the 3 Y. pestis strains tested. Sandwich dot ELISA using monospecific antisera as a capture antibody and a monoclonal antibody, and Yps3 as a revealing antibody had a high level of specificity in detecting Y. pseudotuberculosis antigens.

Molecular Relatedness between Isolates Yersinia pseudotuberculosis from a Patient and an Isolate from Mountain Spring Water

A 40-yr-old buddhist monk was admitted to the hospital with abdominal pain, fever, and confusion. He had a history of drinking untreated mountain spring water in his temple, and experienced the above symptoms for several days before admission. In past medical history, he had suffered from hepatic cirrhosis. Yersinia pseudotuberculosis was isolated from his blood and ascitic fluid. The mountain spring water that he had ingested was cultivated and Y. pseudotuberculosis was also isolated. For identification of pathogenic Y. pseudotuberculosis, each isolate from the three sources (blood, ascitic fluid, and drinking water) was also analysed for the inv gene for Y. pseudotuberculosis and the virF gene for virulent plasmid by PCR. All strains were positive for both the virF and the inv genes and also positive for autoagglutination test. For relationship study, each isolate from the three sources was also analysed with serotyping and restriction endonuclease analysis of virulence plasmid DNA (REAP) using BamHI. All belonged to the serotype 4b and REAP pattern D. Thus, all these findings supported that the mountain spring water was the source of the Y. pseudotuberculosis infection in this case.

Fatores contemporâneos que afetam a incidência de patógenos causadores de doenças de origem alimentar / Contemporary factors that affect the incidence of foodborne pathogens

A modernizaçäo da sociedade e os avanços na tecnologia alimentar resultaram em mudanças na formulaçäo, produçäo e distribuiçäo dos alimentos criando, consequentemente, situaçöes anteriormente imprevistas, que se constituem em novos desafios microbiológicos. Inúmeros fatores inter-relacionados contribuem para agravar o risco de doenças de origem alimentar, dentre os quais se destacam, as inovaçöes na agropecuária, mudanças nos hábitos alimentares e o aumento da suscetibilidade a infecçöes. Esta revisäo focaliza a situaçäo atual das doenças de origem alimentar, considerando os principais agentes, os fatores de risco e a importância da conscientizaçäo da populaçäo para a prevençäo de surtos epidêmicos.

Yersinia pseudotuberculosis Septicemia Report of a Case

Yersinia pseudotuberculosis is known to cause septicemia, mesenteric lymphadenitis enteritis and erythema nodosum. Most of the infections were found in European countries, but none in Korea ti11 now. For the first time in Korea Y. pseudotuberculosis was isolated form a 51-year-old ma1e with liver cirrhosis. The patient showed chills, abdominal pain and diarrhea followed by a comatose state. The organism was isolated from both blood and peritoneal fluid. The isolation and identification were difficult as the organism grew slowly and many of the characteristics were similar to other enteric bacilli. The isolate was susceptible to all antibiotics tested in vitro, but our chemotherapy with ampicillin and kanamycin did not save the patient's life.