Rastreabilidade das condições higiênico-sanitárias do processo de fabricação do queijo de coalho artesanal.

Abstract

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.