Parasitological diagnosis in ornamental freshwater fish from different fish farmers of five Brazilian states.

This study aimed to search parasites in 333 ornamental fish from five Brazilian states (Ceará, Minas Gerais, São Paulo, Paraná and Santa Catarina). Fish were sent from eight farms located in the municipalities of Fortaleza, Patrocínio do Muriaé, São Francisco do Glória, Cascavel, Timbó, Iguape, Jacareí and Mairinque. All fish received anesthesia earlier to euthanasia procedures. After the search for parasites, it was verified that 70.6% (235/333) of fishes were infected by at least one type of parasite, being 12 types of parasites identified: monogeneans, digenean metacercariae, cestodes, nematodes, Lernaea cyprinacea, trichodinids, Piscinoodinium pillulare, Ichthyophthirius multifiliis, diplomonad flagellates, Ichthyobodo sp., Chilodonella sp., and Tetrahymena sp. The proportion of infected fishes among the farms is compared through statistical tests, besides, animal handling adopted in each farm is also discussed. The importance of ensuring fish health in order to make the ornamental freshwater fish industry economically viable and reduce losses in production is highlighted.

Isolation of potential zoonotic Mycobacterium spp. from diseased freshwater angelfish (Pterophyllum scalare) from an aquarium.

Nontuberculous mycobacteria infection is one of the most common chronic bacterial diseases in ornamental aquarium fish and appears to be directly related to stressful husbandry practices. Furthermore, it also represents zoonotic potential. Here we present the isolation and characterization of non-tuberculous mycobacteria from diseased freshwater angelfish (Pterophyllum scalare) in São Paulo, Brazil. Nine discarded breeding females with signs of disease were evaluated. The fish exhibited lethargy, loss of appetite, cachexia, skin ulcers, and exophthalmia. At necropsy, four fishes presented macroscopic granulomas in the spleen. Mycobacterium chelonae, M. fortuitum, M. gordonae, M. intracellulare and M. peregrinum were isolated and identified by hsp65 PCR restriction analysis. Histopathological analysis revealed microscopic lesions compatible with mycobacteriosis, and Mycobacterium bacillus were observed by Ziehl-Neelsen stain. Notably, all Mycobacterium species identified in this study have already been reported in human patients; therefore, diseased animals may be a source of infection for people who handle fish and aquariums.

Methods for thermal inactivation of pathogens in mozzarella: a comparison between stretching and pasteurization / Métodos para inativação térmica de patógenos em mozarela: comparação entre filagem e pasteurização

This study aimed to evaluate the efficiency of stretching in the reduction of pathogens when compared to milk pasteurization, the official method to ensure safe cheese production. Whole buffalo milk was contaminated with Mycobacterium fortuitum, Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus. Part of the milk was used in mozzarella production and the other part was submitted to holder pasteurization. Pathogens were quantified before and after thermal processing (mozzarella stretching and milk pasteurization). Pasteurization and stretching led to the following reductions in log cycles, respectively: 4.0 and 6.3 for Mycobacterium sp.; 6.0 and 8.4 for Listeria sp.; >6.8 and 4.5 for Staphylococcus sp.; and >8.2 and 7.5 for Salmonella sp.

Este estudo teve como objetivo avaliar a eficácia da filagem na redução de patógenos,em comparação coma pasteurizaçãodo leite, que é o método oficialpara garantir aprodução de queijos seguros. Leite de búfala integral foi contaminado com Mycobacterium fortuitum, Listeria monocytogenes, Salmonella typhimurium e Staphylococcus aureus. Parte desse leite foi empregada na fabricação da mozarela e outra parte foi submetida à pasteurização lenta. Os patógenosforam quantificadosantes e após os processos térmicos (filagem da mozarela e pasteurização do leite). As reduções, em ciclos logarítmicos, causadas pela pasteurização e pela filagem, respectivamente, foram: 4,0 e 6,3 de Mycobacterium sp., 6,0 e 8,4 de Listeria sp., >6,8 e 4,5 de Staphylococcus sp. e >8,2 e 7,5 de Salmonella sp.

Identification of Mycobacterium avium genotypes with distinctive traits by combination of IS1245-based restriction fragment length polymorphism and restriction analysis of hsp65.

One-hundred eight Mycobacterium avium isolates from pigs, humans, birds, and bovines were typed by the IS1245-based restriction fragment length polymorphism (RFLP) method and PCR-restriction enzyme analysis (PRA) of hsp65. Nine clusters of isolates showing more than 80% similarity in their RFLP profiles were detected. The largest cluster (cluster B) included 32 of 79 pig isolates (40.5%), 3 of 25 human isolates (12%), and 1 of 2 bovine isolates, comprising 33% of all isolates. The second largest cluster (cluster A) included 18 pig isolates (22.8%) and 6 human isolates (24%). Six smaller clusters included six pig isolates (clusters C and D), four and two human isolates (clusters E and F, respectively), two pig isolates (cluster I), and two pig isolates plus one bovine isolate and the avian purified protein derivative strain (cluster H). Cluster G represented the "bird-type" profile and included the bird isolate in this series, one pig isolate, plus reference strain R13. PRA revealed four allelic variants. Seventy-seven isolates were identified as M. avium PRA variant I, 24 were identified as M. avium PRA variant II, 6 were identified as M. avium PRA variant III, and 1 was identified as M. avium PRA variant IV. Except for three isolates from cluster B, each of the RFLP clusters was associated with a single PRA pattern. Isolates with unique (nonclustered) RFLP profiles were distributed between PRA variants I and II, and there was one unique isolate of PRA variant IV. These observations are consistent with divergent evolution within M. avium, resulting in the emergence of distinct lineages with particular competence to infect animals and humans.

Identification of two novel Mycobacterium avium allelic variants in pig and human isolates from Brazil by PCR-restriction enzyme analysis.

Mycobacterium avium complex (MAC) is composed of environmental mycobacteria found widely in soil, water, and aerosols that can cause disease in animals and humans, especially disseminated infections in AIDS patients. MAC consists of two closely related species, M. avium and M. intracellulare, and may also include other, less-defined groups. The precise differentiation of MAC species is a fundamental step in epidemiological studies and for the evaluation of possible reservoirs for MAC infection in humans and animals. In this study, which included 111 pig and 26 clinical MAC isolates, two novel allelic M. avium PCR-restriction enzyme analysis (PRA) variants were identified, differing from the M. avium PRA prototype in the HaeIII digestion pattern. Mutations in HaeIII sites were confirmed by DNA sequencing. Identification of these isolates as M. avium was confirmed by PCR with DT1-DT6 and IS1245 primers, nucleic acid hybridization with the AccuProbe system, 16S ribosomal DNA sequencing, and biochemical tests. The characterization of M. avium PRA variants can be useful in the elucidation of factors involved in mycobacterial virulence and routes of infection and also has diagnostic significance, since they can be misidentified as M. simiae II and M. kansasii I if the PRA method is used in the clinical laboratory for identification of mycobacteria.

Isolamento de Niocardia asteroides de lesäo granulomatosa em carne bovina / Isolation of Niocardia asteroides of granulomatous lesions in bovine meat

Os produtos de origem animal desempenham importante papel na alimentaçäo humana devido ao seu valor protéico, porém deve-se evitar que sirvam de vias de transmissäo de doenças aos seus consumidores. A interaçäo dinâmica entre o serviço de inspeçäo de carnes e as técnicas laboratoriais diagnósticas deve ser mantida na profilaxia das doenças veiculadas por alimentos. Relata um caso de contaminaçäo bacteriana de tecido muscular de bovino destinado ao consumo humano. Macroscopicamente observaram-se lesöes granulomatosas semelhantes àquelas associadas à tuberculose. Colheu-se 2,0g do material e, utilizando-se a técnica de descontaminaçäo de Petroff (OPS 1993), tentou-se o isolamento de Mycobacterium ssp, a partir dos meios de Lowenstein-Jensen e de Stonebrink-Lesslie a 37§C. Outro fragmento foi colhido e fixado em formol a 10 por cento para exame histopatológico. Após sete dias de cultivo observou-se no meio de Lowenstein-Jensen a presença de colônias rugosas com pigmento alaranjado. No exame bacteriológico observou-se micélios que se fragmentavam formando bacilos Gram positivos. No corte histológico, corado em Hematoxilina-Eosina, verificaram-se estruturas granulomatosas. Foi identificado Nocardia asteroides a partir das colônias isoladas. Estes microrganismos säo considerados saprófitas, telúricos capazes de causar infecçöes inicialmente pulmonares, disseminando-se posteriormente para outros órgäos, como: rins, baço, fígado, cérebro e ocasionalmente de desenvolver micetomas. Sem auxílio de profissionais integrados e capacitados para diagnosticar esta patologia, haveria maior risco para o consumidor. Conclui-se, portanto, que a crescente integraçäo entre os profissionais de saúde pública e saúde animal permite o aprimoramento das práticas diagnósticas e de prevençäo das zoonoses.

[Tuberculosis lymphadenitis in slaughtered swine from the State of São Paulo, Brazil: microscopic histopathology and demonstration of mycobacteria]. / Linfadenites tuberculóides em suínos abatidos no estado de São Paulo, Brasil: aspectos macroscópicos histopatológicos e pesquisa de micobactérias.

INTRODUCTION: As the occurrence of tuberculosis lymphadenitis in swine constitutes a public health risk, especially in immunosuppressed individuals, the distribution of tuberculoid lesions and the presence of microbacteria in lymph nodes and hepatic and muscular tissue in swine for slaughter, in the State of S. Paulo, SP (Brazil), in the period from 1993 to 1994, were studied. MATERIAL E METHOD: Tuberculous lesions in 60 carcasses of swine, slaughtered at abattoirs during the period 1993-1994, were studied. When macroscopic lesions were observed, a representative sampling of lymph nodes, hepatic and muscular tissues (masseter and diaphragm) were examined using histopathological and microbacterial isolation techniques. RESULTS: The macroscopic lesions were found predominantly in mesenteric lymph nodes. The MAI Complex (M.avium-intracellulare) was isolated mainly in the carcass group that showed lesions and was not found in the control group. Microorganisms of the MAI Complex were not isolated from hepatic and muscular tissues. CONCLUSION: No clear relationship between the type of mycobacteria isolated and the macroscopic lesions observed during the carcass inspection was found.