Effects of chelating calcium in cryopreservation extender on frozen-thawed dog semen / [Efeitos da quelação de cálcio no meio de criopreservação no sêmen descongelado de cão]

We evaluated the effect of reducing free calcium in the cryopreservation medium, using the calcium chelator ethylene diamine tetracetic acid (EDTA) at 0.3% and 0.5% concentrations. Three male mixed breed dogs were subjected to semen collection by digital manipulation (n=16). Each ejaculate was divided in three aliquots, and each one was diluted in TRIS-glucose-egg yolk extender with 6% glycerol and 0.5% Equex STM Paste® (TGE, control); and added with 0.3% EDTA (EDTA 0.3) or 0.5% EDTA (EDTA 0.5). Calcium concentration reduced in EDTA 0.3 and all the calcium ions were chelated in EDTA 0.5. The EDTA addition did not affect sperm morphology or plasma membrane integrity; however, by removing all free calcium (EDTA 0.5), the sperm motility reduced (64.7% in TGE and 45% in EDTA 0.5; p<0.05). Acrosome integrity and sperm binding ability were not improved by calcium chelation. The failure to prevent the premature AR may explain why sperm longevity was not affected by calcium removal. Thus, the partial or complete calcium removal, through EDTA addition, is not able to prevent acrosomal damage or premature acrosomal reaction, and therefore does not improve the dog sperm binding ability.(AU)

Avaliou-se o efeito da redução do cálcio livre no meio de congelamento, usando-se o quelante de cálcio etilenodiaminotetracético (EDTA) a 0,3% e 0,5%. Três cães machos sem raça definida foram submetidos à coleta de sêmen por manipulação digital (n=16). Cada ejaculado foi diluído em diluidor controle com TRIS-glicose - gema de ovo (TGE, controle), ou em diluidor TGE enriquecido com 0,3% (EDTA 0,3) ou 0,5% de EDTA (EDTA 0,5). A concentração de cálcio reduziu no meio EDTA 0,3, e todos os íons de cálcio foram quelados no meio EDTA 0,5. A adição do EDTA e a consequente quelação do cálcio não afetaram a morfologia espermática ou a integridade da membrana plasmática, no entanto, ao remover todo o cálcio do meio (EDTA 0,5), a motilidade espermática se reduziu (64,7% no TGE e 45% no EDTA 0,5; P<0,05). A integridade do acrossoma e a capacidade de ligação do espermatozoide não melhoraram com a quelação do cálcio. Apesar da influência da concentração de cálcio sobre a motilidade espermática após o descongelamento, a falha em prever a reação acrossomal prematura pode explicar por que a longevidade espermática não foi afetada pela remoção do cálcio no meio. Dessa forma, a remoção parcial ou total do cálcio, por meio da adição de EDTA, não é capaz de prevenir o dano no acrossoma ou a reação acrossomal prematura e, portanto, não aumenta a capacidade do espermatozoide de se ligar ao oócito.(AU)

Desordens hematológicas em onças-pardas (Puma concolor; Linnaeus, 1771) infectadas por Citauxzoon felis / [Hematological disorders in Brown jaguars (Puma concolor; Linnaeus 1771) infected by Citauxzoon felis]

Este estudo objetivou descrever o aspecto hematológico de seis onças-pardas (Puma concolor) infectadas pelo Cytauxzoon felis. Os seis casos de infecção foram identificados durante o manejo sanitário de 11 animais de um centro de reabilitação de animais silvestres. Estruturas compatíveis com piroplasmídeos foram observadas durante a avaliação do esfregaço sanguíneo e confirmadas como Cytauxzoon felis pela técnica de PCR. A análise estatística demonstrou diferença significativa (P<0,05) no número absoluto dos linfócitos entre os grupos dos animais infectados e não infectados. Assim, expressivas alterações hematológicas e bioquímicas entre os grupos investigados alertam para a dificuldade de identificação de onças-pardas infectadas por C. felis, apoiada apenas em exames de rotina, bem como para o risco, sobretudo, da reintrodução desses animais na natureza.(AU)

This Cytauxzoon felis by the PCR technique. Statistical analysis showed a significant difference is study aimed to describe the hematological appearance of six puma (puma concolor) infected with cytauxzoon felis. The six cases of infection were identified during the sanitary management of 11 animals from a wild animal rehabilitation center. Piroplasmid compatible structures were observed during the blood smear evaluation and confirmed as (P<0.05) in the absolute number of lymphocytes between the groups of infected and uninfected animals. Thus expressive hematological and biochemical alterations between the groups investigated alert to the difficulty of identifying infected brown jaguars by C. felis, supported only by routine examinations, and the risk especially when aiming at the reintroduction of these animals in the wild.(AU)

Avaliação da qualidade microbiológica do queijo-de-minas artesanal do Serro-MG / Microbiological quality of artisanal minas cheese, manufactured in the region of Serro-MG

Avaliaram-se a qualidade microbiológica do queijo-de-minas artesanal do Serro e a variação da microbiota do queijo recém-fabricado e no último dia de validade, após estocagem sob as condições recomendadas no rótulo. Foram analisadas 40 amostras de queijo, sendo 20 analisadas frescas, imediatamente após a coleta, e 20 no último dia de validade, após estocagem em câmara fria a 10ºC. As amostras foram analisadas para coliformes a 35ºC e 45ºC, Staphylococcus coagulase positiva, Salmonella spp. e Listeria monocytogenes. Trinta e sete amostras (92,5 por cento) encontraram-se impróprias para o consumo humano, de acordo com os parâmetros estabelecidos pela resolução: RDC ANVISA nº12/01, sendo a principal causa de condenação a contagem de Staphylococcus coagulase positiva. Nenhuma das amostras analisadas apresentou contaminação por Salmonella sp. ou Listeria monocytogenes. As contagens de Staphylococcus coagulase positiva dos queijos frescos foram mais altas (P<0,01) que as dos queijos analisados no último dia da validade

The microbiological quality of artisanal minas cheese produced in the region of Serro-MG was evaluated. Cheeses were analyzed fresh and at the end of the shelf life, under specific storage conditions (30 days, 10ºC). Forty samples of minas cheese were analyzed, twenty still fresh, and 20 after storage at 10ºC. The microbiological analyses were counts of fecal and total coliforms, and Staphylococcus coagulase positive; and search of Salmonella sp. and Listeria monocytogenes. Twenty-seven samples (92.5 percent) were in disagreement with the Brazilian Standard, Resolution nº 12 of January 12, 2001, from ANVISA (National Health Surveillance Agency), and Staphylococcus coagulase positive was the major contaminant. No sample was contaminated by Salmonella sp. or Listeria monocytogenes. During the storage, there was a decrease of countings in the population of Staphylococcus coagulase positive (P<0.01)

Mortalidade por "septicemia dos peixes tropicais" em tilápias criadas em consorciação com suínos

A rural property was observed for one year to obtain seasonal observations of Edwardsiella septicemia in tilapias (Oreochromis sp.) from an integrated fish farm (using pig excrements as food). Fifty tilapias on summer, forty-five on autumn, fifty-five on winter and one hundred and five on spring were collected and evaluated microbiologically for the presence of Edwardsiella tarda. Samples of the external surface (skin, gills and fin), intestines and muscle were analyzed. Fishes were transported alive to the laboratory, and maintained for five days for observation. The external signs observed were opacity of cornea, respiratory difficulty, disordered swimming, nodules on gills, hemorrhagic lesions under the skin and fins and incoordination of the posterior part of the body. In general, after three days of observation, the tilapias died. At necropsy, ascites with the distension of the celomatic cavity, hemorrhagic and necrotic lesions in the liver, spleen and kidneys were found. Hemorrhagic enteritis was observed in the gut tube. Edwardsiella tarda isolated presented the following biochemical profile: negative urea, sucrose, phenylalanine desaminase and malonate utilization, and positive indole, methyl red, H2S, motility, glucose with gas production and lysine decarboxylase. The mortality rates were 69.9% on spring, 63.6% on winter, 48.0% on summer and 40.0% on autumn, highest rates occurring on spring and winter (P 0,05).

Com o objetivo de estudar a variação sazonal da septicemia dos peixes tropicais provocada por Edwardsiella tarda foram observadas 50 tilápias (Oreochromis sp.) no verão, 45 no outono, 55 no inverno e 105 na primavera. Os peixes doentes apresentavam opacidade de córnea, dificuldade respiratória, nado desordenado, nódulos nas brânquias e lesões hemorrágicas na pele e nadadeiras. Após três dias de observação as tilápias morriam naturalmente. Nas necrópsias foi encontrado ascite com distensão da cavidade celomática e lesões hemorrágicas e necróticas no fígado, baço e rins. No interior do tubo digestivo observou-se enterite hemorrágica. Isolou-se Edwardsiella tarda a partir de provas positivas para H2S, lisina, indol, motilidade e glicose com gás. Os índices de mortalidade foram: na primavera 69,9%, no inverno 63,6%, no verão 48,0% e no outono 40,0%.

Hematological changes induced by Bothrops jararaca venom in dogs

The mechanism of consumption coagulopathy observed in cases of human envenomation by Bothrops jararaca is well established. However, this mechanism may vary according to the animal species studied. In order to study both the clinical and laboratory aspcts of bothropic envenomation in dogs, a sublethal defibrinating dose of venom (100 µg/kg) was given intravenously. A coagulopathy similar to that observed in humans - including fibronogen depletion, consumption of factors II, X, V and antithrombin III, and moderate thrombocytopenia -was observed. The presence of circulatin activated platelets was also noted. Neutrophilic leukocytosis, lymphopenia, and monocytosis occurred at different times. Erythrocytic values remained normal in dogs treated with B. jararaca venom compared with those treated with saline alone. The erythrocyte sedimentation rate fell rapidly after venom administration and this fall was correlated logarithmically with fibrinogen concentration. Since the effect of envenomation in dogs is similar to that in humans, it was concluded that the dog can be used as a good animal model for studying human venom-induced coagulopathy