Diagnóstico das condições higienicossanitárias do processo de fabricação de queijo de coalho no sertão paraibano

As condições gerais e adequações quanto às Boas Práticas de Fabricação (BPF) foram averiguadas em oito queijarias artesanais de queijo de coalho no sertão Paraibano, com o principal objetivo de investigar as conformidades e não conformidades relativas a sua implementação. Para tanto, usou-se como critério avaliativo um checklist baseado na Resolução RDC n? 275 da ANVISAlMS. Os resultados mostraram que das oito queijarias, apenas uma classificou-se como de baixo risco, ou seja, com mais de 76% de adequações. Nos itens edificações e instalações, controle de pragas e da água, higiene dos manipuladores, ausência de tratamento térmico da matéria-prima e documentação, as queijarias D e E apresentaram os maiores níveis de não conformidade. Conclui-se que as queijarias artesanais apresentam um alto risco para veiculação de Doenças Transmissíveis por Alimentos e, se faz necessária a adoção das BPF pelas queijarias e uma maior fiscalização pelos órgãos oficiais, pois as inadequações observadas podem comprometer a qualidade dos produtos e a segurança dos consumidores.(AU)

The general conditions and adequacies regarding the Good Manufacturing Practices (GMP) were verified in eight artisanal curdled cheese factories with the objective to investigate the conformities and non-conformities related to its implementation. Therefore, it was used as evaluation criteria, a checklist based on the Resolution - RDC number 27J of ANVISA/MS. The results showed that from the eight evaluated cheese factories, only one was classified as low risk, that is, with more than 76% of adequacies. On the edification and facilities aspects, plague and water control, handlers' hygiene, lack of thermal treatment of raw material and documents, cheese factories D and E showed the highest levels of non-conformity. It can be concluded that artisanal cheese factories present a high risk for the propagate of Foodborne Diseases, so it is necessary the adoption of GMP by the cheese factories and a stricter supervision made by the official bodies since the verified inadequacies could compromise the quality of the products as well as the safety of consumers.(AU)

RASTREABILIDADE DAS CONDIÇÕES HIGIÊNICO-SANITÁRIAS DO PROCESSO DE FABRICAÇÃO DO QUEIJO DE COALHO ARTESANAL

Esta tese é composta por três artigos, que estão divididos em capítulos. No primeiro capítulo foi abordado as condições gerais e adequações quanto às Boas Práticas de Fabricação (BPF) de oito queijarias artesanais de queijo de coalho, com o principal objetivo de investigar as conformidades e não conformidades relativas a sua implementação. Para tanto, usou-se como critério avaliativo um cheklist baseado na Resolução RDC nº 275 da ANVISA/MS, onde os resultados mostraram que das oito queijarias, apenas uma classificou-se como de baixo risco, ou seja, com mais de 76% de adequações. As principais não conformidades foram: instalações, controle de pragas e da água, higiene dos manipuladores, ausência de tratamento térmico da matéria prima e documentação. O segundo capítulo relata os principais pontos de contaminação do queijo de coalho em queijarias artesanais do Sertão paraibano, avaliando a qualidade higiênica do processamento através da quantificação de coliformes, mesófilos, Escherichia coli e identificação de Staphylococcus spp. Em amostras de leite, queijos, swabs do tanque de fabricação, mesa, fôrma e mãos dos manipuladores, além da água utilizada na fabricação e limpeza dos equipamentos, onde os resultados demonstraram que o leite cru de todas as queijarias apresentaram alta contaminação microbiológica, estando em desacordo com a IN 62 do Ministério da Agricultura, Pecuária e Abastecimento. A superfície da mesa, tanque, fôrma e mãos dos manipuladores também apresentaram altas contagens para todos os micro-organismos. No queijo detectou-se coliformes a 30º C, termotolerantes e E. coli. As contagens de Staphylococcus spp. apresentaram acima de 1,3 x 105UFC/g, podendo favorecer a produção de enterotoxinas sob condições adequadas e os principais agentes isolados nas amostras foram S. aureus, S. epidermidis, S. haemolyticus, S. lugdunensis. O leite apresentou S. aureus em todas as queijarias, porem, no queijo houve predomínio de espécies coagulase negativa, sendo também encontradas nos utensílios e mãos dos manipuladores. O terceiro capítulo aborda a identificação dos genes FemA, Coa com sequenciamento das amostras positivas para esse gene e de enterotoxinas clássicas nas 51 cepas de Staphylococcus spp isoladas a partir de amostras de leite, queijo de coalho e utensílios obtidas em três queijarias artesanais. De acordo com a identificação das espécies, das 51 cepas obtidas, 16 foram identificadas como S. aureus, porém, todas as amostras de Staphylococcus spp. foram submetidas a técnica de PCR para presença do gene femA e coa, onde houve amplificação do gene femA em 40 amostras e apenas 20 amostras, cujos tamanhos variaram de aproximadamente 500 a 900 pb. Na pesquisa das enterotoxinas, nenhum dos isolados analisados possuía os genes sec e see. Houve apenas a amplificação para o gene sea e seb em uma mesma amostra e sed em outra. As amostras positivas para o gene Coa foram submetidas ao Blast e exibiram grande homologia com sequencias codificadoras do gene da estafilocoagulase de S. aureus depositadas no NCBI. Foi construída uma árvore filogenética baseada nas sequencias, onde foi apresentada a relação entre as amostras. As cepas oriundas do leite, queijo, mãos dos manipuladores da queijaria A apresentaram bastante similaridade, sugerindo que o leite cru e as mãos contribuíram para a contaminação do queijo. O mesmo ocorreu com as amostras das mãos e tanque da queijaria C. Este estudo mostrou que a identificação genotípica de Staphylococcus spp. pode ser feita pela amplificação do gene femA e do gene Coa, com apresentação de grande polimorfismo genético. A identificação do gene para enterotoxinas mostrou-se útil para avaliar um possível perfil dos S. aureus e o sequenciamento confirmou os pontos de contaminação bacteriana no processamento de fabricação do queijo de coalho.

This thesis is composed by three articles which are divided into chapters. On the first chapter, it was discussed the general conditions and adequacies regarding the Good Manufacturing Practices (GMP) of eight artisanal curdled cheese factories and which the main objective was to investigate the conformities and non-conformities related to its implementation. Therefore, it was used, as evaluation criteria, a checklist based on the resolution RDC number 275 of ANVISA/MS, and the results showed that from the eight evaluated cheese factories, only one was classified as low risk, that is with more than 76% of adequacies. The main non-conformities were: facilities, water and plague control, handlers hygiene, lack of thermal treatment of raw material and documents. The second chapter reports the main contamination spots of curdled cheese in artisanal cheese factories in the sertão of Paraíba, evaluating the hygienic quality of processing through the quantification of coliforms, mesophilic, Escherichia coli and identification of Staphylococcus spp. In milk samples, cheese, swabs from the fabrication tank, table, mold and handlers hands, and also on water used in the fabrication and cleaning of the equipment, the results showed that the raw milk from all the cheese factories was highly contaminated with microorganisms, therefore being in discordance with the Normative Instruction (IN) 62 of the Ministry of Agriculture, Livestock and Food Supply. The surface of the table, mold and handlers hands also showed high rate of all microorganisms. On the cheese, it was detected thermotolerant coliforms at 30º C as well as E. coli. The counts for Staphylococcus spp. showed more than 1,3 x 105UFC/g, which favors the production of enterotoxins under adequate conditions and the main isolated agents on the samples were S. aureus, S. epidermidis, S. haemolyticus, S. lugdunensis. The milk indicated S. aureus in all cheese factories. However, on the cheese there was a predominance of coagulase-negative specimens, which were also found in utensils and handlers hands. The third chapter discusses the identification of FemA, Coa genes, with sequencing of positive samples for this gene as well as the classical enterotoxins on the 51 strains of Staphylococcus spp isolated from milk samples, curdled cheese and utensils from three artisanal cheese factories. According to the identification of the specimens, of the 51 obtained strains, 16 were identified as S. aureus. However, all Staphylococcus spp. samples were submitted to PCR technique to detect the presence of femA and coa gene, where there was amplification of the femA gene in 40 samples and only in 20 samples the size varied from approximately 500 to 900 pb. In the research of the enterotoxins, none of the analyzed which were isolated had the sec and see genes. There was only the amplification for the sea and seb gene on the same sample and for the sed gene in another. The samples which were positive for the Coa gene were submitted to Blast and showed great equivalence with coding sequences of the Staphylococci coagulase of S. aureus stored in NCBI. A phylogenetic tree was built based on the sequences which presented relation with samples. The strains originated from milk, cheese, handlers hands showed great similarity, suggesting that the raw milk and the hands contributed for the cheese contamination. The same situation occurred with the samples from hands and from tank of cheese factory C. This study showed that the genotypic identification of Staphylococcus spp. can be done by the amplification of femA and Coa genes, with abundant genetic polymorphism. The identification of the gene for enterotoxins resulted to be useful to evaluate a possible S. aureus profile and the sequencing confirmed the bacterial contamination spots in the processing of curdled cheese fabrication.

Ocorrência de leite ácido e de resíduos de antimicrobianos no leite cru entregue em laticínio na região do Vale do Taquari, RS, Brasil / Occurrence of Acidity and Antimicrobial Residues in Raw Milk Delivered at a Dairy Plant in Vale do Taquari, RS, Brazil

Milk is considered one of the most important sources of nutrients. It contains proteins, fat acids, vitamins and minerals. Its activity has a significant socio-economic impact in all over the world and in Brazil. The presence of bacterial inhibitors in milk for human consumption is a great concern for the dairy products industry and public health. With regard to the production of dairy products, mainly cheeses and fermented milks, this affects the growth of starter cultures in ripening processes of cheese and fermented milk. The risks for consumer health are represented by allergic reactions, which are usually manifested as urticarias, dermatitides or rhinitis and bronchial asthma, and that can result in anaphylactic shock in susceptible individuals. Another risk to be considered is the consumption by pregnant women of milk containing high levels of antimicrobial residues, considering the teratogenic potential presented by some of them. By the other hand, milk acidity is one of the biggest problems encountered at milk processing plants and, for that reason, the control of the storage temperature of products and the adoption of adequate hygienic practices must be prioritized. The objective of this study was to verify the incidence of acidity and antimicrobial residues in milk, at a dairy factory from Vale do Taquari, RS.

Ocorrência de leite ácido e de resíduos de antimicrobianos no leite cru entregue em laticínio na região do Vale do Taquari, RS, Brasil / Occurrence of Acidity and Antimicrobial Residues in Raw Milk Delivered at a Dairy Plant in Vale do Taquari, RS, Brazil

Milk is considered one of the most important sources of nutrients. It contains proteins, fat acids, vitamins and minerals. Its activity has a significant socio-economic impact in all over the world and in Brazil. The presence of bacterial inhibitors in milk for human consumption is a great concern for the dairy products industry and public health. With regard to the production of dairy products, mainly cheeses and fermented milks, this affects the growth of starter cultures in ripening processes of cheese and fermented milk. The risks for consumer health are represented by allergic reactions, which are usually manifested as urticarias, dermatitides or rhinitis and bronchial asthma, and that can result in anaphylactic shock in susceptible individuals. Another risk to be considered is the consumption by pregnant women of milk containing high levels of antimicrobial residues, considering the teratogenic potential presented by some of them. By the other hand, milk acidity is one of the biggest problems encountered at milk processing plants and, for that reason, the control of the storage temperature of products and the adoption of adequate hygienic practices must be prioritized. The objective of this study was to verify the incidence of acidity and antimicrobial residues in milk, at a dairy factory from Vale do Taquari, RS.(AU)

Evaluation of hygienic-sanitary conditions of the minas frescal and mozzarella cheese produced in Morrinhos city -GO / Avaliação das condições higiênico-sanitárias do queijo minas frescal e mussarela produzido na cidade de Morrinhos – GO

Microbiological analyses were performed in 60 samples of Minas Frescal cheese and 60 samples of Mozzarella cheese. Samples were collected and produced in a dairy factory in Morrinhos-GO from April to September, for quantitative analysis of total coliforms, fecal coliforms, Staphylococcus, mold and yeast. Concerning the presence of total coliforms the highest score to Minas Frescal cheese which was impossible to count; on the other hand, to Mozzarella cheese the highest score was 1.7.101 UFC/g, to fecal coliforms analyses, Minas Frescal Cheese presented under 1.0.101 UFC/g in all samples and the Mozzarella type showed 1.7.101 UFC/g. Analyses of Staphylococcus demonstrated the higher score for Minas Frescal which was 3.5.103 UFC/g; whereas Mozzarella cheese score was under 1.0.101 UFC/g; mold and yeast score was 3.4.102 UFC/g to Minas Frescal and 9.3.102 UFC/g to Mozzarella. Thus, results showed different scores for both cheeses. Minas Frescal cheese demonstrated higher indexes than Mozzarella when compared total coliforms and Staphylococcus, and Mozzarella presented higher fecal coliforms and moulds and yeasts analyses. According to described above, it is important to verify the hygienic-sanitary care in the foods manipulation.

Foram realizadas análises microbiológicas em 60 amostras de queijo Minas Frescal e 60 amostras de queijo Mussarela produzidas em Morrinhos - GO. As amostras foram coletadas em um laticínio de Morrinhos entre os meses de abril a setembro, avaliando de forma quantitativa, coliformes totais, coliformes fecais, Staphylococcus, bolores e leveduras. Com relação a presença de coliformes totais o resultado mais elevado para Minas frescal foi de incontáveis UFC/g, enquanto para o queijo Mussarela foi 1,7.101 UFC/g, nas análises de coliformes fecais o queijo Minas Frescal apresentou resultados 1,0.101 UFC/g em todas as amostras e o tipo Mussarela apresentou 1,7.101 UFC/g. Com relação as análises de Staphylococcus o índice mais elevado para o queijo Minas Frescal foi 3,5.103 UFC/g, enquanto para o queijo Mussarela obtivemos 1,0.101 UFC/g em todas as amostras, os resultados de bolores e leveduras foi 3,4.102 UFC/g para o Minas Frescal e 9,3.102 UFC/g para as amostras do queijo Mussarela. Os resultados obtidos apresentaram alterações para os dois tipos de queijo, Minas Frescal apresentou índices superiores quando comparados coliformes totais e Staphylococcus, o queijo Mussarela apresentou resultados mais elevados com as análises de coliformes fecais e bolores e leveduras. Palavras-chave: Queijo Minas Frescal; Queijo Mussarela; Avaliação Microbiológica.

Evaluation of hygienic-sanitary conditions of the minas frescal and mozzarella cheese produced in Morrinhos city -GO / Avaliação das condições higiênico-sanitárias do queijo minas frescal e mussarela produzido na cidade de Morrinhos GO

Microbiological analyses were performed in 60 samples of Minas Frescal cheese and 60 samples of Mozzarella cheese. Samples were collected and produced in a dairy factory in Morrinhos-GO from April to September, for quantitative analysis of total coliforms, fecal coliforms, Staphylococcus, mold and yeast. Concerning the presence of total coliforms the highest score to Minas Frescal cheese which was impossible to count; on the other hand, to Mozzarella cheese the highest score was 1.7.101 UFC/g, to fecal coliforms analyses, Minas Frescal Cheese presented under 1.0.101 UFC/g in all samples and the Mozzarella type showed 1.7.101 UFC/g. Analyses of Staphylococcus demonstrated the higher score for Minas Frescal which was 3.5.103 UFC/g; whereas Mozzarella cheese score was under 1.0.101 UFC/g; mold and yeast score was 3.4.102 UFC/g to Minas Frescal and 9.3.102 UFC/g to Mozzarella. Thus, results showed different scores for both cheeses. Minas Frescal cheese demonstrated higher indexes than Mozzarella when compared total coliforms and Staphylococcus, and Mozzarella presented higher fecal coliforms and moulds and yeasts analyses. According to described above, it is important to verify the hygienic-sanitary care in the foods manipulation.

Foram realizadas análises microbiológicas em 60 amostras de queijo Minas Frescal e 60 amostras de queijo Mussarela produzidas em Morrinhos - GO. As amostras foram coletadas em um laticínio de Morrinhos entre os meses de abril a setembro, avaliando de forma quantitativa, coliformes totais, coliformes fecais, Staphylococcus, bolores e leveduras. Com relação a presença de coliformes totais o resultado mais elevado para Minas frescal foi de incontáveis UFC/g, enquanto para o queijo Mussarela foi 1,7.101 UFC/g, nas análises de coliformes fecais o queijo Minas Frescal apresentou resultados 1,0.101 UFC/g em todas as amostras e o tipo Mussarela apresentou 1,7.101 UFC/g. Com relação as análises de Staphylococcus o índice mais elevado para o queijo Minas Frescal foi 3,5.103 UFC/g, enquanto para o queijo Mussarela obtivemos 1,0.101 UFC/g em todas as amostras, os resultados de bolores e leveduras foi 3,4.102 UFC/g para o Minas Frescal e 9,3.102 UFC/g para as amostras do queijo Mussarela. Os resultados obtidos apresentaram alterações para os dois tipos de queijo, Minas Frescal apresentou índices superiores quando comparados coliformes totais e Staphylococcus, o queijo Mussarela apresentou resultados mais elevados com as análises de coliformes fecais e bolores e leveduras. Palavras-chave: Queijo Minas Frescal; Queijo Mussarela; Avaliação Microbiológica.