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Background and Aim: Flemish cattle in Brazil are on the brink of extinction and are found only in one herd in Lages, Santa Catarina State. This study aimed to uncover the reasons for the recurring abortions in the Flemish cattle herd. Materials and Methods: Seventeen Flemish fetuses underwent postmortem examinations, with samples collected for histopathology and microbiology culture tests, polymerase chain reaction (PCR) test for Neospora caninum, and reverse transcription-PCR (RT-PCR) test for bovine viral diarrhea virus (BVDV) from 2015 to 2020. Results: Of the 17 fetuses, N. caninum was the most common diagnosis and was found in 88% (15/17). One fetus (5.8%) had a coinfection with N. caninum and Citrobacter amalonaticus, leading to fibrinonecrotic pericarditis. All fetuses tested negative for BVDV by RT-PCR. Of the 107 dams tested by indirect immunofluorescence assay, 26 (25.2%) were anti-N. caninum seropositive, with 17 (65.4%) aborting and 5 (19.2%) having estrus repetition. Reverse transcription-PCR results showed that 9 (8.4%) of the serum samples collected from dams tested positive, which tested follow-up test 3 months later, indicating a BVDV transient infection. The factors that contributed to neosporosis included dogs’ access to pastures and improper disposal of fetal remains, which made it easier for dogs to consume them. Conclusion: This study warns the occurrence of N. caninum as a cause of reproductive disorders that can lead to abortion in the studied Flemish cattle herd.

The serrano artisanal cheese is a typical product from South region of Brazil, which is produced by skilled cheesemakers using raw milk. The contamination of this food by Escherichia coli has a great impact on public health, since it could threat the consumers’ health. The study evaluated the presence of virulence genes, antimicrobial susceptibility profiles and bofilm-production ability of Escherichia coli isolates obtained from raw milk and artisanal cheese produced in Southern Brazil. A total of 117 isolates of E. coli were characterized by multiplex PCR to detect the following virulence genes: eae for enteropatogenic E. coli (EPEC), lt and st for enterotoxigenic E. coli (ETEC), stx for shiga toxin-producing E. coli (STEC), stx and eae for enterohemorrhagic E. coli (EHEC), ipaH for enteroinvasive E. coli (EIEC) and aggR for enteroaggregative E. coli (EAEC). In addition, antimicrobial susceptibility profile to 22 antimicrobial agents was also performed by disk diffusion method, and we searched for extended-spectrum beta-lactamases (ESBL) and/or carbapenemase- producing isolates. Isolates that were positive for ESBL and carbapenemase were further investigated for the presence of the genes: blaTEM, blaSHV, blaOXA, blaCTX-M, for ESBL and blaOXA-48 for carbapenemase. Further, isolates had their ability to form biofilms investigated by the red Congo agar method. Virulence genes of E. coli were identified in 21.37% of the tested isolates, which were classified as EPEC (the most prevalent pathotype) and ETEC or EAEC. Ten (8.55%) of the total studied E. coli isolates revealed a multidrug-resistant profile, since they were resistant to three or more antimicrobial classes; whereas four isolates (3.42%) were classified as ESBL-producers and showed the presence of blaTEM gene. None of the isolates exhibited carbapenemase activity nor did they carry carbapenemase genes. From the total of E. coli isolates, 79 (67.52%) were considered potential biofilm producers. These results address a serious public health issue, since artisanal cheeses pose a risk to consumers’ health, since may be sources of dissemination of diarrheogenic E. coli, that can cause from subclinical to severe and fatal infections in children and adults, and also emphasize the need to improve adaptations/adjustments in the manufacturing processes of these products.

O queijo artesanal serrano (QAS) é produzido a partir de leite cru, tornando-se um meio propício para a multiplicação de micro-organismos patogênicos. O processo de maturação envolve fatores físicos, químicos e microbiológicos que desfavorecem o desenvolvimento de patógenos. Diante disso, a legislação exige um período mínimo de 60 dias de maturação, porém, de acordo com a IN nº 30, de 7 de agosto de 2013 e com o Decreto nº 9.013, de 29 de março de 2017, pode-se considerar a possibilidade de reduzir esse tempo, desde que estudos científicos comprovem que não há alteração da inocuidade do produto. O objetivo do trabalho foi avaliar a qualidade microbiológica de QAS, em 19 queijarias da Serra Catarinense, com 14, 21, 28 e 35 dias de maturação e verificar qual período atenderia aos limites microbiológicos. Foram objetos de estudo 76 amostras de QAS, 19 amostras de água e 19 amostras de leite cru. Os micro-organismos pesquisados seguiram a Portaria nº 146, de 07 de março de 1996. A pesquisa e quantificação de coliformes a 30°C, E. coli e Staphylococcus coagulase-positiva, seguiram protocolos de Silva et al. (2010), utilizando Petrifilm™. A pesquisa de Salmonella spp. seguiu Silva et al. (2010) e a pesquisa de Listeria monocytogenese e a quantificação de aeróbios mesófilos seguiram a Instrução Normativa nº 30, de 26 de junho de 2018. Para avaliação da potabilidade da água foi realizada a técnica do Número Mais Provável (NMP), seguindo a IN nº 30/2018 e a Portaria de Consolidação nº 5, de 28 de setembro de 2017. Os resultados passaram por teste de Análise de Variância e Regressão Linear do pacote estatístico SAS com significância (p < 0.05). Nesse estudo, não foi possível determinar um período de maturação seguro para comercialização. Aos 35 dias de maturação, 14 propriedades ainda apresentaram quantificações acima do limite para os micro-organismos descritos na legislação. Em uma das propriedades, foi detectada a presença de L. monocytogenes nos quatro períodos avaliados, demonstrando que a maturação não foi eficiente para eliminar o patógeno. A qualidade microbiológica do leite não foi satisfatória, sendo que 63,16% das amostras estavam fora dos padrões. Com relação à potabilidade da água das queijarias, 52,63% das amostras analisadas apresentaram resultados insatisfatórios. A pesquisa indicou heterogeneidade nos padrões higiênico-sanitários das queijarias, demonstrando que são necessárias capacitações em Boas Práticas Agropecuárias e Boas Práticas de Fabricação, apoio de instituições, fiscalização periódica e comprometimento dos produtores para padronização do processo.

The serrano artisan cheese produced from raw milk of dairy cattle is a typical product of high-altitude farms in the states of Santa Catarina and Rio Grande do Sul. However, marketing of the cheeses occurs illegally because they lack the minimum maturation period required for cheese produced from raw milk. The production of artisan cheeses is required to follow strict hygiene standards. This study aimed to test the quality and safety of cheeses that were produced in 31 farms of the Serrana region in Santa Catarina after 14 and 28 days of maturation. Coliform count was measured at 35 °C, and presence of other microorganisms such as Escherichia coli, Staphylococcus, Listeria spp., and Salmonella spp. were also tested. Fat and protein percentages, acidity, salt content, and humidity were also evaluated. Data were subjected to statistical analyses using the SAS® software. After 14 and 28 days of maturation, 74.19% (23/31) and 64.52% (20/31) of samples, respectively, showed higher numbers of coliforms at 35 °C than those permissible by law. Higher than permissible numbers of E. coli were observed in 45.16% (14/31) and 48.39% (15/31) of the samples analyzed after 14 and 28 days of maturation, respectively. Coagulase-positive staphylococci values above 103 CFU/g were observed in 54.84% (17/31) and 51.61% (16/31) of cheese samples after 14 and 28 days of maturation, respectively. Contamination with Salmonella spp. was not detected. However, Listeria monocytogenes serovar 4b was isolated in 3.23% (1/31) and 6.45% (2/31) of samples after 14 and 28 days of maturation, respectively. The results of humidity tests classified the cheese samples into three categories: low, medium, and high humidity. Semi fat cheeses were predominant in both maturation periods, although the samples were classified in thin, semi fat, and fat cheeses. The main variations in the compositions of analyzed samples occurred for salt and acidity levels. The maturation process has not proven to be effective in reducing microbiological contamination to compliance levels. Considering the heterogeneity of the analyzed cheese samples, the frequency of non-conformities with respect to microorganisms and pathogens present in the samples, this study indicates the necessity to improve the Serrano artisan cheese production system through adoption of good manufacturing practice measures.

Among multiresistant Gram-positive microorganisms, stands out methicillin-resistant Staphylococcus (MRS), an opportunistic pathogen associated with hospital acquired and community infections reported in medicine and large increase in reports of veterinary medicine. In veterinary medicine, numerous reports regarding several species of animals have been described. MRS is intrinsically resistant to all ?-lactam drugs. In veterinary medicine, numerous reports regarding several species of animals have been described, but Staphylococcus aureus with intermediate resistance and resistant to vancomycin (VISA/VRSA) has not yet been reported in veterinary medicine, still need further study. Staphylococcus spp. are also related to antimicrobial resistance of macrolides, lincosamides, and streptogramin B (MLSB) group, that has the same mechanism of action, although the drugs belong to different classes. In veterinary medicine, clindamycin (lincosamide class) is widely used for skin infections, wounds, bone infections, pneumonia, infections of the oral cavity, and infections caused by anaerobic bacteria, besides being used for treatments of MRS infections. Enterococcus is another resistant Gram-positive microorganism, from which vancomycin-resistant enterococci (VREs) are the most important strains. There are several reports of VREs in veterinary medicine due the use of a similar antimicrobial (avoparcin) in livestock; therefore this group of microorganisms has now acquired great prominence since vancomycin is considered as the last resort for the treatment of MRS and Enterococcus associated with nosocomial infections in humans. The biggest problem these microorganisms and their resistance mechanisms cause is related to its huge impact on public health due to the increasing close contact between animals and humans. The objective of this review was to identify the main Gram-positive microorganisms associated with animals, describing their mechanisms of action that lead to antimicrobial resistance, as well as their impact on public health through their zoonotic and anthropozoonotic potential.

Vacuum-packed and frozen of ostrich (Struthio camelus), alligator (Caiman latirostris) and wild boar (Sus scrofa) meat samples were obtained in an authorized commercial store in Maringá/Paraná. Of each kind meat were analyzed 6 samples, where were studied counts of mesophilic aerobic bacteria, aerobic bacteria psichrophilic, coliforms, Escherichia coli and Staphylococcus spp., and protein and fat analysis. The results of aerobic mesophilic bacteria ranged between