Esta pesquisa ocorreu em uma comunidade rural do Mato Grosso (34 propriedades) e foi dividida em quatro etapas: 1) Palestra sobre “A qualidade e segurança microbiológica de leite e derivados”; 2) Questionário (16 produtores) sobre o manejo de ordenha e produção de lácteos; 3) Visita técnica em cinco propriedades produtoras de leite e derivados e coleta de amostras; 4) Pesquisa (10 amostras) de micro-organismos indicadores de qualidade e deteriorantes. Na palestra (etapa 1) notou-se uma lacuna na compreensão da relevância das boas práticas na qualidade e segurança alimentar. O questionário (etapa 2) apontou para ausência de assistência técnica em 81% das propriedades; produção e comercialização de queijos (com leite cru) (56%) e leite (37%); higienização das mãos com água e sabão (87%) e pré e pós dipping (56%) como rotina na ordenha. Das Boas Práticas de Produção (BPP), 67% realizavam a higienização das mãos e 50% usavam vestimentas exclusivas/ adequadas. Na etapa 3, foi constatada a presença de uma infraestrutura inadequada e a comercialização dos lácteos ocorria na comunidade e feiras livres. Os resultados microbiológicos indicaram falhas na higiene de produção, contaminação de origem fecal e risco de intoxicação estafilocócica. Pseudomonas spp. (149) apresentaram predomínio da capacidade lipolítica, com alto potencial deteriorante lipoproteolítico e lipolítico em leite e queijos, respectivamente. Deve-se considerar a necessidade de um olhar mais atento à comunidade, com o objetivo de proporcionar apoio técnico que assegure a qualidade e a segurança dos lácteos produzidos, contribuindo para maior lucratividade e a sustentabilidade dos produtores na região. Palavras-chave: Segurança Alimentar. Saúde Única. Leite. Queijo. Abstract This research was carried out in a rural community in Mato Grosso (34 farms) and was divided into four phases: 1) Lecture on "The Quality and Microbiological Safety of Milk and Dairy Products"; 2) Questionnaire (16 farmers) on milking management and dairy production; 3) Technical visit to five dairy farms and sampling; 4) Survey (10 samples) of quality indicators and spoilage microorganisms. The lecture (stage 1) revealed a gap in understanding the importance of good practices for food quality and safety. The questionnaire (stage 2) showed the absence of technical assistance in 81% of the farms, the production and marketing of cheese (with raw milk) (56%) and milk (37%), hand hygiene with soap and water (87%) and pre and post dipping (56%) as routine milking. Of the good production practices (GPP), 67% practiced hand hygiene and 50% wore exclusive/appropriate clothing. In stage 3, inadequate infrastructure was found and dairy products were sold in the community and at street markets. The microbiological results showed deficiencies in production hygiene, fecal contamination, and the risk of staphylococcal poisoning. Pseudomonas spp. (149) showed a predominance of lipolytic capacity, with high lipoproteolytic and lipolytic spoilage potential in milk and cheese, respectively. The need for a closer look at the community should be considered, to provide technical support to ensure the quality and safety of the dairy products produced, contributing to greater profitability and sustainability for farmers in the region. Keywords: Food Safety. One Health. Milk. Cheese.
Salmonella spp. is a pathogen associated with foodborne infections, mainly in foods of animal origin. In this context, the present study investigated the occurrence of Salmonella serotypes, genotypes and the antimicrobial resistance profiles of strains in fresh beef produced in Mato Grosso, Brazil.A total of 107 samples from 13 different slaughterhouses in the Mato Grosso were analyzed. Suggestive Salmonella spp. colonies detected during the biochemical screening were submitted to DNA extraction, and hilA gene amplification was used for the PCR reaction. Antimicrobial resistance analyses were performed using 17 antimicrobial agents from eight different classes by the disk diffusion method. Strains exhibiting multiple drug resistances were submitted to PCR genotyping based on repetitive sequences (rep-PCR), using a commercial semiautomatic DiversiLab® system.A total of 5.6% (6/107) of the samples tested positive by the conventional method and were confirmed by PCR, namely two S. Akuafo, two non-typable Salmonella enterica strains, one Salmonella O:16 serovar, and one S. Schwarzengrund. The antimicrobial resistance profiles indicated resistance to gentamicin (30%), tetracycline, nitrofurantoin, and trimethoprim + sulfamethoxazole (16%). Genotyping indicated a 70% difference between S. Schwarzengrund and the non-typable Salmonella strains. No genetic similarities were observed between the six Salmonella isolates based on rep-PCR, including two S. Akuafo.The results obtained herein corroborate that Salmonella serovar Schwarzengrund is commonly isolated in animal products in the state of Mato Grosso, Brazil, also highlighting the presence of two unusual Salmonella serovars in beef (Akuafo and O:16).
ABSTRACT: Brazil is the largest exporter of beef of the world and Mato Grosso State is the highest beef producer in this country. To maintain product competitiveness and market expansion, sanitary hygienic control of the entire process is indispensable to ensure the attainment of harmless beef and quality. The objective of this study was to evaluate the hygienic sanitary conditions of vacuum-packed beef produced by establishments qualified for export in the state of Mato Grosso, Brazil. A total of 60 samples were submitted to coliforms counts at 35°C and at 45°C and E. coli. The mean contamination by at 35°C and coliforms at 45°C were 3,1 x 102MPN/g and 7.7MPN/g respectively. The presence of E. coli was verified in five samples, representing an occurrence of 8.3% (5/60), and Salmonella spp. in 5% (03/60) of the analyzed samples. The MPN (Most Probable Number) average of coliforms at 35°C and 45°C are in accordance to national and international legislation; however, the presence of Samonella spp., E. coli in some sample indicates a low risk of occurrence of salmonellosis and colibacillosis transmitted by the evaluated beef. However, transmission risk of these diseases cannot be ruled out, since the presence of E. coli does not depend on the amount of coliforms and national legal standards established for the group of thermotolerant coliforms.
A pandemia da COVID-19 causada pelo vírus SARS-CoV-2 trouxe desafios adicionais ao Serviço de Alimentação e Nutrição (SAN), pois, além de atender as condições higiênicas sanitárias dos alimentos, as empresas agora precisam incorporar novas práticas que visem preservar a saúde das pessoas. Nesse contexto, esse estudo buscou realizar uma revisão da literatura sobre as recomendações técnicas de boas práticas de fabricação de alimentos relacionadas com a prevenção da COVID-19 e elaborar uma lista de verificações (check-list) para facilitar a identificação de falhas, avaliar risco de transmissão da COVID-19 nos serviços de alimentação e orientar as adaptações dos manuais de boas práticas de fabricação. Com base nas notas técnicas vigentes no Brasil e baseadas nas diretrizes da Organização Mundial de Saúde (OMS), foi elaborado um questionário contendo 100 perguntas nas diversas categorias do setor de produção de alimentos. Dessa maneira, entendemos que medidas de biossegurança devem ser adotadas a partir da atualização dos manuais de boas práticas de fabricação nos serviços de alimentação, e, para apoiar essa ação, propomos a utilização do checklist em apêndice para identificar as inconformidades relacionadas a prevenção da COVID-19.
Introduction: Mycobacterium bovis is a member of the M. tuberculosis complex (MTC), a group of species with a high genetic homology. Is a causative agent of bovine tuberculosis (BTB), zoonosis that causes economic and public-health problems in many countries. Non-pasteurized milk and milk produtcts are an important transmission route, resulting in extra-lung presentation of the illness. The definitive diagnosis is realized by traditional culture and biochemical methods. This procedure is cumbersome and time-consuming. We evaluated a PCR assay for the direct detection of M. bovis DNA in naturally contaminated milk, using primers that were previously tested and proven reliable to target the IS6110 element. Methodology: Milk previously seeded with M. bovis was used as the starting material, for padronization of the technique. The DNA was extracted from milk samples by a modification of a QIAamp Blood and Tissue Kit (Qiagen). Primers INS1 (5'-CGTGAGGGCATCGAGGTGGC-3') and INS2 (5'-GCGTAGGCGTCGGTGACAAA-3') to IS6110 element were used to amplification of region genomic present only in members of Mycobacterium tuberculosis Complex (MTC). A total of 150 adult cows which were reactive to cervical comparative intradermal tuberculin test (CITT) from various Brazilian dairy herd with a persistent previous history of bovine tuberculosis were used in this study. Milk samples were collected and subjected to bacteriological culture and PCR assay. Results e discussion: The PCR assay allowed us to detect BTB in artificially contaminated milk, with a detection limit of 100 CFU/mL, and was also able to detect the bacillus in 50% (75/150) of samples from naturally infected animals. Although we detected by PCR the DNA of Bacillus in the milk of 75 cows (50%), the confirmation of viability was observed only for 38 animals (25%). The results confirmed that milk was a potential source of infection for cattle and also to humans, and thus a public health risk consumption of milk and dairy products do not treated. Conclusion: The PCR assay allowed the detection of M. bovis DNA in artificially contaminated milk and also showed the ability to detect the bacilli of bovine tuberculosis in naturally infected milkand could be an alternative option for implementation in epidemiological studies of BTB transmission and to prevent contaminated milk from entering the food supply.
Abstract Historically, Escherichia coli is among the most studied organisms and serves as the basis for understanding many fundamental biochemical and genetic concepts. In addition, it displays 9 pathogenesis groups, with the Shiga toxin‐producing (STEC) group being the main representative regarding foodborne pathogenesis. Its typical characteristic is the presence of 2 distinct toxins and variants: stx1 ( stx1a , stx1c , and stx1d ), and stx2 ( stx2a , stx2b , stx2c , stx2d , stx2e , stx2f , and stx2g ). The main challenge regarding the study of E. coli is the standardization of a high sensitivity method including all pathotypes, that allows for enrichment of STEC cells and a decrease of background microbiota. The ability of some E. coli cells belonging to other pathogenic groups, such as O104:H4, to acquire genes unique to the STEC group, increases the pathogenic power and the risk of new outbreaks related to these bacteria. In addition, animals with a high concentration of pathogenic E. coli cells present in feces (above 10 4 CFU/g), designated as supershedding animals, may be the primary transmission factor among ruminants. Therefore, the purpose of this review is to address pathogenicity factors and the importance of supershedding animals in the transmission of this pathogen, discussing the main methods currently applied, to focus on the occurrence of STEC in beef.
Introduction: Salmonella spp. is often associated with food borne outbreaks in several countries around the world, causing serious public health problems. The diagnosis of this species is usually performed by classical bacteriological cultures and serological testing, which are laborious and time-consuming methods. Aiming at an effective and rapid diagnosis, molecular techniques, such as PCR, allow for the rapid and specific detection of the pathogen. In this context, the present study aimed to perform a PCR standardization for the rapid and specific identification of Salmonella spp.. Methods: Primers designed for hilA gene, previously described in the literature, hilA2-F (5''-CTGCCGCAGTGTTAAGGATA-3'') and hilA2-R (5''-CTGTCGCCTTAATCGCATGT-3'') were selected and used for the internal standardization of optimal Salmonella spp.-specific PCR conditions. Thermal lysis was used to extract DNA from several Salmonella strains, used as positive reference controls, from Escherichia coli, Staphylococcus aureus, Enterobacter aerogenes, Klebsiella pneumoniae, used as negative controls, and from 16 environmental Salmonella spp. strains. The amplification conditions were adjusted in order to obtain the specific amplification of the fragment (496 bp) from the smallest possible amount of DNA. The resulting PCR products were analyzed by electrophoresis on ultrapure 1.5% agarose gels stained with ethidium bromide (10 mg mL-1). Results and Discussion: The optimum conditions determined in this study were: initial denaturation at 94°C for 5 min followed by 30 cycles at 94°C for 1 min, 58°C for 1 min, 72°C for 1 min and a final extension at 72°C for 10 min, using a mixture of 5 Andamp;#956;L reaction buffer (Invitrogen), 0.2 mM dNTP (Fermentas), 1.5 U recombinant Taq polymerase (Taq Platinum® - Invitrogen), 5 mM MgCl2 (Invitrogen) and 25 pmol of each primer (Invitrogen). The PCR amplified the expected fragment (496 bp) in these conditions only for the positive control and the previously identified environmental Salmonella spp. strains, showing 100% sensitivity and specificity and a limit of detection of 50 ng target DNA. Conclusion: It was possible to identify isolated Salmonella spp. colonies with 100% sensitivity and specificity using PCR with the optimum conditions standardized in the present study, reducing pathogen detection time to 5 hours.
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