Determination of pathogens and contamination indicatorsin fresh pork sausages in the state of São Paulo / Determinação de patógenos e indicadores de contaminação em linguiças frescais de origem suína no estado de São Paulo

The highest cost of beef and chicken meat justifies higher consumption of fresh sausage products, especially linguiça, due to their easy preparation and affordability for consumers, making necessary an evaluation of sanitary hygienic conditions of these products. The objective was to investigate the presence of pathogens such as Salmonella spp., coagulase-positive Staphylococcus, thermotolerant coliforms, Escherichia coli and Campylobacter spp. in artisanal and inspected fresh pork sausages. It was found that 12% (6/50) of the artisanal sausage samples were contaminated with Salmonellaspp., 58% (29/50) presented coagulase-positive Staphylococcus levels above the acceptable limits for consumption and 76% (38/50) presented thermotolerant coliform levels above the acceptable limits. In sausage samples produced under inspected conditions, 6% (3/50) were contaminated with Salmonella spp., 24% (12/50) presented thermotolerant coliform levels above the acceptable limits, 2% (1/50) presented enteropathogenic E. coli. None samples showed coagulase-positive Staphylococcus counts above the limits, or presence of Campylobacter spp. Sensitizing traders and consumers about the importance of inspection service in food of animal origin is urgent for a sanitary acceptable production, since foodborne diseases continue to be a public health problem.

O alto custo das carnes de frango e bovina justifica maior consumo de produtos embutidos frescais, especialmente a linguiça, devido ao preço acessível ao consumidor e fácil preparo, tornando necessário estudos para avaliar as condições higiênico-sanitárias. Os objetivos foram verificar a presença de Salmonella spp., Staphylococcus coagulase positiva, coliformes termotolerantes, Escherichia colipatogênicas e Campylobacter spp. em linguiças suínas tipo frescal artesanais e fiscalizadas. Os resultados obtidos em linguiças artesanais foram de 12% (6/50) contaminadas com Salmonella spp., 58% (29/50) de Staphylococcus coagulase positiva e 76% (38/50) nas quantificações de coliformes termotolerantes, dados que apresentam níveis de contaminação superiores aos da legislação vigente. Nas linguiças produzidas sob fiscalização detectou-se 6% (3/50) de contaminação por Salmonella spp.; 24% (12/50) de quantificação de coliformes termotolerantes acima dos limites aceitáveis, e 2% (1/50) de E. coli enteropatogênica. Nenhuma amostra apresentou contagens de Staphylococcus coagulase positiva fora dos padrões, ou contaminação por Campylobacter spp. Sensibilizar comerciantes e consumidores sobre a importância do serviço de inspeção em alimentos de origem animal é premente para que haja uma produção sanitária aceitável, pois doenças transmitidas por alimentos continuam sendo um problema à saúde pública.

Survey for Leptospira and Brucella in Amazonian manatees, Amazon river dolphins, and a tucuxi in the Brazilian Amazon.

Leptospirosis and brucellosis are zoonotic diseases with global distributions that represent severe hazards to humans and animals. We investigated exposure to Leptospira spp. and Brucella spp. in samples from Amazonian manatees Trichechus inunguis, Amazon river dolphins Inia geoffrensis, and a tucuxi Sotalia fluviatilis. The animals were free-ranging or undergoing in situ rehabilitation in the mid-Solimões River region, Brazilian Amazon. Serum samples from 19 Amazonian manatees were tested by microscopic agglutination test, Rose Bengal test, and 2-mercaptoethanol Brucella agglutination test. Antibodies against Leptospira spp. were detected in 63% of the manatees tested and serovar Patoc was considered the infecting serovar in all positive samples. Titers were generally low, indicating chronic exposure, but higher titers indicative of an active infection were detected in 3 animals. Anti-Brucella spp. antibodies were not detected. Tissue and/or body fluid samples from 12 Amazon river dolphins, a tucuxi, and 2 Amazonian manatees were investigated by multiplex PCR and bacteriology for Leptospira spp. and Brucella spp. All samples were negative. However, Enterococcus faecalis was isolated from uterine fluid, lymph node, and lung of 3 Amazon river dolphins. Bacillus spp. were isolated from milk and synovial fluid from 2 Amazon river dolphins and from a milk sample from 1 Amazonian manatee. Knowledge of the pathogens present in Amazonian manatees, Amazon river dolphins, and tucuxis is of great relevance to species conservation and environmental health. Although no clinical signs were noted, further research is needed to elucidate the clinical relevance of infection by Leptospira sp. serovar Patoc in Amazonian aquatic mammals.

Detection of virulence factors in coagulase-negative Staphylococcus spp. strains isolated from Emmental cheese / Detecção de fatores de virulência em cepas de Staphylococcus spp. coagulase negativos isoladas de queijo Emmental

Food prepared with products derived from animals are involved in most cases of staphylococcal poisoning; therefore, the research of Staphylococcus spp. in Emmental cheese is more applicable. The objective of this study was to identify coagulase-negative Staphylococcus spp. (CNS) in cheese using biochemical and molecular techniques to detect the presence of nine genes responsible for the production of enterotoxins. From 180 samples analyzed, 204 CNS strains were obtained and identified as being 46 (22.6%) S. saprophyticus strains, 27 (13.2%) S. hominis spp. hominis strains, 22 (10.8%) S. sciuri strains, 21 (10.3%) S. xylosus strains, 19 (9.3%) S. epidermidis strains, 19 (9.3%) S. haemolyticus strains, 17 (8.3%) S. lentus strains, 17 (8.3%) S. warneri strains, 11 (5.4%) S. equorum strains and 5 (2.5%) S. cohnni . Using the PCRm protocol, 14 (6.9%) strains with the presence of the genes on the enterotoxin E (SEE)11 (78.6%), J (SEJ) 1 (7%), C (SEC) 1 (7%) and I (SEI) 1 (7%) were detected. Based on the results, the type of package is not interfered of growth and isolated that Staphylococcus spp. in cheese. It was observed that bacteria capacity to produce coagulase cannot be understood as an indicative of enterotoxigenicity; therefore, the CNS should be considered as a target of importance in the epidemiology of staphylococcal intoxications. It can be concluded that CNS need to be included in bacterial foodborne disease research, since the genes responsible for the production of toxins were detected and none of the studied samples presented Staphylococcus spp. counting above the limits allowed by legislation.(AU)

Os alimentos preparados com produtos de origem animal são os mais envolvidos em casos de intoxicação alimentar estafilocócica; portanto a pesquisa do Staphylococcus spp. em queijos tipo Emmental é relevante. O objetivo foi isolar e identificar espécies de Staphylococcus coagulase negativas (CNS)de queijo Emmental acondicionado em vários tipos de embalagem, por meio de técnicas bacteriológicas e bioquímicas e detectar, por PCR, a presença de nove genes responsáveis pela produção de enterotoxinas. Das 180 amostras, foram isoladas 204 cepas de CNS, que foram identificadas por provas bioquímicas como: 46 (22,6%) S. saprophyticus, 27 (13,2%) S. hominis spp. hominis, 22 (10,8%) S. sciuri, 21 (10,3%) S. xylosus, 19 (9,3%) S. epidermidis , 19 (9,3%) S. haemolyticus , 17 (8,3%) S. lentus , 17 (8,3%) S. warneri , 11(5,4%) S. equorum e 5 (2,5%) S. cohnii . Na PCR multiplex, em 14 (6,9%) isolados foi detectada a presença dos genes para enterotoxina E (SEE), em 11 (78,6%) J (SEJ), em 1 (7%) C (SEC) e em 1 (7%) I (SEI). Com base nos resultados, o tipo de embalagem não interferiu na multiplicação dos Staphylococcus spp. isolados dos queijos. Neste estudo, verificou-se que a capacidade para a produção de coagulase pela bactéria não pode ser concebida como indicativa de enterotoxigenicidade, portanto devem-se considerar os CNS como objeto de importância na epidemiologia das intoxicações estafilocócicas, fazendo-se necessária a atenção com relação à pesquisa dos CNS nos alimentos, uma vez que foram detectados genes responsáveis pela produção de toxinas, e nenhuma das amostras apresentou contagem para Staphylococcus spp. acima do limite permitido pela legislação.(AU)

Causes of equine abortion, stillbirth, and perinatal mortality in Brazil / Causas de abortamento, natimortalidade e mortalidade em equinos no Brasil

Abortion and complications in reproduction are important causes of economic loss in horse breeding. Studies of its causal agents can help to identify the primary pathogens or other factors involved and define appropriate measures to reduce its occurrence. This research aimed to investigate the primary causes of equine abortion, stillbirth, and perinatal mortality in regions of Brazil. Tissue from aborted fetuses, stillbirths, neonates and foals submitted to the Biological Institute of São Paulo, Brazil, from January 2010 to July 2013 were processed for viral and bacterial isolation, polymerase chain reaction (PCR), histology, and immunohistochemistry. Bacterial infection was the primary detected cause of abortion, found in 16 of the 53 animals submitted for bacterial analysis followed by viruses analysis in 2 of 105 animals, and noninfectious causes (neonatal isoerythrolysis) in 2 of 105 animals. Fungi were found in a single sample of 53 tested. The most frequent bacteria recovered were Escherichia coli, Enterobacter aerogenes, combined E. coli and Streptococcus spp., Staphylococcus spp., and Bacillus spp. The following agents were each observed in a single sample: Arcanobacterium pyogenes, Streptococcus spp., Corynebacterium spp., Actinobacillus spp., and Rhodococcus equi. The predominant identification of fecal and other opportunistic bacteria as opposed to pathogens commonly associated with equine abortion, such as Leptospira spp. and equine herpesvirus type 1 (EHV-1), suggests the need of improving hygiene management of breeding mares to prevent bacterial infection that may cause fetal loss, stillbirth, and perinatal mortality.(AU)

Abortamento e complicações na reprodução são importantes causas de perda econômica na equideocultura. Estudos dos agentes causais podem ajudar a identificar patógenos ou outros fatores envolvidos e definir medidas apropriadas para reduzir sua ocorrência. Esta pesquisa investigou as causas primárias de aborto, natimortalidade e mortalidade perinatal em equinos de diversas regiões do Brasil. Tecidos de fetos abortados, natimortos e potros submetidos ao Instituto Biológico de São Paulo, Brasil, no período de janeiro de 2010 a julho de 2013, foram processados por meio de técnicas de isolamento viral e bacteriano, PCR, histologia e imuno-histoquímica. Infecção bacteriana foi a causa mais detectada, encontrada em 16 de 53 amostras submetidas à análise bacteriana, seguida de causa viral em 2 de 105 amostras, e causas não infecciosas (isoeritrólise neonatal) em 2 de 105 amostras. Fungo foi encontrado em uma única amostra de 53 testadas. As bactérias isoladas mais frequentemente foram Escherichia coli, Enterobacter aerogenes, E. coli associada a Streptococcus spp., Staphylococcus spp. associado a Bacillus spp. Os seguintes agentes foram observados em uma única amostra cada: Arcanobacterium pyogenes, Streptococcus spp., Corynebacterium spp., Actinobacillus spp. e Rhodococcus equi. A identificação predominante de bactérias fecais e outras bactérias oportunistas, ao invés de outros patógenos comumente associados a quadros de abortamento equino, tais como Leptospira spp. e Herpesvírus equino tipo 1, sugere a necessidade de maior atenção no manejo higiênico das éguas em reprodução, a fim de prevenir infecções bacterianas que possam causar perda fetal, natimortalidade e mortalidade perinatal.(AU)

Demodex phylloides infection in swine reared in a peri-urban family farm located on the outskirts of the Metropolitan Region of São Paulo, Brazil.

This paper reports the occurrence of porcine demodicosis caused by the mite Demodex phylloides in hogs reared in a peri-urban family farm located in Francisco Morato, a municipality of the Metropolitan Region of São Paulo, capital city of the state of São Paulo, Brazil. In a parcel of forty Landrace - Large White cross hogs, approximately four months old, four animals presented severe skin lesions in the form of small nodules over their entire body, especially in the periocular region, snout, lower abdomen and flanks. Two hogs had to be euthanized for animal welfare reasons, which enabled post-mortem examination. Skin scrapings revealed eggs, larvae, nymphs and adults of D. phylloides. Purulent subcutaneous nodules with intense parasitic folliculitis and intense perifollicular inflammatory reaction were present. Enterobacteria and coagulase-positive Staphylococcus spp. were isolated from the skin pustules. Necropsy revealed milk spotted liver, enteritis and enlargement of mesenteric lymph nodes. Protozoa (Eimeria spp. and Balantidium sp.), helminth eggs (Ascaris suum, Trichuris suis and strongyles) and Brachyspira spp. were found in faeces. Staphylococcus spp. and enterobacteria were isolated from internal organs. All remaining hogs were treated with ivermectin at a daily oral dose of 0.45g/kg of feed, during seven days. Fifteen days after treatment, remission of symptoms was observed in the surviving animals with demodicosis; absence of mites was confirmed by skin scraping examinations. The hogs were reared under poor environmental, nutritional and sanitary conditions, resulting in multimorbidity and immunosuppression. Severe clinical porcine demodicosis was triggered when the animals were castrated. Family pig farmers had been suffering economic losses due to the stunted growth of the herd. In addition to that, the lesions found on the skin and in the internal organs would result in condemnation of meat and viscera for human consumption. As part of a Public Policies Project, farm facilities were renovated with governmental aid, while family farmers received training. Good management practices and biosecurity measures were introduced in the herd. Educative policies and financial support were important to guide family pig farmers towards better husbandry practices, allowing them to raise healthy hogs in compliance with market demands.

Ocorrência de anticorpos anti-Brucella ovis em ovinos com histórico de distúrbios reprodutivos no estado de São Paulo, Brasil / Occurrence of antibodies anti-Brucella ovis in ovine with history of reproductive disorders in São Paulo, Brazil

Com base no histórico de distúrbios reprodutivos, foram realizados inquéritos sorológicos e tentativas de isolamento de Brucella ovis em 28 propriedades do estado de São Paulo, totalizando 294 ovinos. Os soros colhidos por ocasião das visitas aos estabelecimentos rurais foram submetidos ao teste de fixação do complemento. Durante os exames clínicos, também foram colhidas amostras de materiais biológicos suspeitos para posterior cultivo bacteriológico em meio Brucella ágar, num total de 16 fetos abortados, um útero, seis secundinas, 13 secreções uterinas, seis zaragatoas de muco vaginal, 17 amostras de sêmen e três zaragatoas prepuciais. A proporção de ovinos com histórico de distúrbios reprodutivos e sororreativos para B. ovis foi de 1,7% (5/294), sendo um macho e quatro fêmeas, com títulos variando de 800 UI a 1.600 UI. Quatro rebanhos dos 28 pesquisados apresentaram animais sororreagentes (14,3%). Todos os cultivos das amostras biológicas foram negativos, inclusive sêmen e órgãos reprodutivos de carneiro da raça Texel, positivo em dois testes de fixação de complemento com intervalo de seis meses. Nos rebanhos de ovinos do estado de São Paulo examinados, não foi possível relacionar os distúrbios reprodutivos à sorologia positiva para B. ovis. No entanto, foi detectada presença de focos, fator de risco para a disseminação da bactéria nos rebanhos.(AU)

Based upon a history of reproductive disorders, a serological survey and attempts to isolate Brucella ovis were performed in 28 farms in the State of São Paulo, in a total of 294 sheep. The sera sampled on the occasion of the visit to the farms were subject to complement fixation. During clinical examination, samples of several suspicious biological materials were collected for further bacterial culture in Brucella agar medium, in a total of 16 aborted foetuses, one uterus, one placenta, 13 uterine discharge, six vaginal swabs, 17 semen samples and three preputial swabs. The proportion of sheep with a history of reproductive disorders and serum reactive for B. ovis was of 1.7% (5/294), including one ram and four ewes, with titres ranging from 800 IU to 1600 IU. Four of the 28 sheep herds surveyed had serum reactive animals (14,3%). All cultures of biological samples were negative, including semen and swabs from the reproductive organs of a Texel ram, positive in two complement fixation tests, six months apart. In the sheep herds surveyed in the State of São Paulo, it was not possible to relate the reproductive disorders to the positive serology for B. ovis, however, foci were detected, representing a risk factor for bacterial dissemination in the herds.(AU)

Brucella canis infection in dogs attended in veterinary clinics from patos, Paraíba state, Brazil

To determine the frequency of anti-Brucella canis antibodies in dogs attended in veterinary clinics from Patos, Paraíba State, Brazil, as well as to identify risk factors and to isolate and identify the agent, 193 dogs were used. Agar gel immunodiffusion test (AGID) was used to detect B. canis antibodies in sera. Isolation of B. canis was carried out in blood and bone marrow from seropositive animals. Six animals tested seropositive in AGID, resulting in a frequency of 3.11 percent. B. canis was isolated from bone marrow of one seropositive animal, with confirmation by PCR. Lack of cleaning of the dog's environment was identified as risk factor (odds ratio = 7.91). This is the first report of isolation of B. canis in dogs from the Northeast region of Brazil.

Brucella canis infection in dogs attended in veterinary clinics from patos, ParabaIBA state, Brazil.

To determine the frequency of anti-Brucella canis antibodies in dogs attended in veterinary clinics from Patos, Paraíba State, Brazil, as well as to identify risk factors and to isolate and identify the agent, 193 dogs were used. Agar gel immunodiffusion test (AGID) was used to detect B. canis antibodies in sera. Isolation of B. canis was carried out in blood and bone marrow from seropositive animals. Six animals tested seropositive in AGID, resulting in a frequency of 3.11%. B. canis was isolated from bone marrow of one seropositive animal, with confirmation by PCR. Lack of cleaning of the dog's environment was identified as risk factor (odds ratio = 7.91). This is the first report of isolation of B. canis in dogs from the Northeast region of Brazil.

Acceptable microorganisms concentration in a semen sample for in vitro embryo production

The aim of this study was to report that the acceptable concentration of microorganisms in a semen sample for insemination may not be safe for an in vitro fertilization procedure. It seems that the semen sample should be completely germ-free, because of the excellent microorganism proliferation condition promoted by the in vitro environment.

O objetivo foi relatar que a concentração de microrganismos aceitável para uma amostra de sêmen utilizada para inseminação pode não ser segura para a realização de fertilização in vitro. Aparentemente a amostra deve ser isenta da presença de contaminantes, pois a condição in vitro promove ambiente favorável para seu crescimento.

Pesquisa de Salmonella spp, coliformes totais, coliformes fecais, mesófilos, em carcaças e cortes de frango / Researches of Salmonella spp, enterobacteriaceae, mesofilos, in carcasses of chicken

A qualidade microbiológica da carne de frango é fundamental para a saúde pública, pois esta carne deve apresentar uma carga bacteriana baixa. Alguns microorganismos presentes em carne de frango podem ocasionar toxinfecções alimentares, especialmente os pertencentes aos gêneros Salmonella. A contagem padrão de bactérias mesófilas é usada como indicador da qualidade higiênica dos alimentos. O índice de coliformes totais avalia condições higiênicas e o de coliformes fecais é empregado como indicador de contaminação fecal. O presente trabalho teve como objetivo verificar a qualidade microbiológica de carcaças e cortes de frango oriundos de um abatedouro avícola localizado no Estado de São Paulo. Foram analisadas 29 amostras de carcaças e 35 cortes de frango, através das seguintes análises microbiológicas: determinação do número mais provável de coliformes totais e fecais, contagem de mesófilos e pesquisa de Salmonella spp. Os resultados obtidos indicaram que em 6 (20,7 por cento) carcaças de frango e 4(11,4 por cento) corte de frango analisados foi isolados Salmonella Enteritidis. Na contagem de coliformes fecais, das 29 amostras de carcaças de frango, 23 (79,3 por cento) apresentaram-se aceitáveis para o consumo humano, 4 (13,8 por cento) apresentaram-se em condições insatisfatórias para o consumo e 2 (6,9 por cento) apresentaram-se impróprias para o consumo. Das 35 amostras de cortes de frango, 26 (74,3 por cento) apresentaram-se aceitáveis para o consumo humano, 8 (22,8 por cento) apresentaram-se em condições insatisfatórias para o consumo e 1 (29 por cento) apresentou-se imprópria para o consumo. Em todas amostras analisadas, a contagem de mesófilos apresentaram-se de acordo com a legislação vigente.