Anderson Carlos Camargo

The aim of this study was to identify virulence and antimicrobial resistance profiles and determine the sequence type (ST) by multilocus sequence typing (MLST) of Salmonella enterica isolates from bovine carcasses from slaughterhouse located in Minas Gerais state, Brazil, and its relationship with bovine isolates obtained on the American continent based on sequence type profile. The MLST results were compared with all Salmonella STs associated with cattle on American continent, and a multi-locus sequence tree (MS tree) was built. Among the 17 S. enterica isolates, five ST profiles identified, and ST10 were the most frequent, grouping seven (41.2%) isolates. The isolates presented 11 different profiles of virulence genes, and six different antibiotics resistance profiles. The survey on Enterobase platform showed 333 Salmonella STs from American continent, grouped into four different clusters. Most of the isolates in the present study (13/17), were concentrated in a single cluster (L4) composed by 74 STs. As a conclusion, five different STs were identified, with ST10 being the most common. The isolates showed great diversity of virulence genes and antibiotics resistance profiles. Most of the isolates of this study were grouped into a single cluster composed by 74 STs formed by bovine isolates obtained on the American continent.

The goals of this study were to evaluate the persistence and the virulence potential of Listeria monocytogenes isolated from beef carcasses obtained in processing facilities in the Southern region of Rio Grande do Sul, Brazil, based on pulsed field gel electrophoresis (PFGE), invasion ability in human colorectal carcinoma cells (HCT-116), InlA expression by western blot (WB) and identification of mutation points in the inlA . PFGE profiles demonstrated that L. monocytogenes isolates were grouped based on their previously identified lineages and serogroups (lineage I: serogroups IIb, n = 2, and IVb, n = 5; lineage II, serogroup IIc, n = 5), isolates with indistinguishable genetic profiles by this method were obtained from different slaughterhouses and sampling steps, with up to 3-year interval. Seven isolates showed high invasion ability (2.4 to 7.4%, lineage I, n = 6, lineage II, n = 1) in HCT and expressed InlA. Five isolates showed low cell invasion ability (0.6 to 1.4%, lineage I, n = 1, lineage II, n = 4) and did not express InlA, and two of them (lineage II, serogroup IIc) presented mutations in inlA leading to a premature stop codon (PMSC) type 19, at position 326 (GAA → TAA). The results demonstrated that most of L. monocytogenes isolates from Lineage I expressed InlA and were the most invasive in HCT indicating their high virulence potential, while most isolates from Lineage II showed attenuated invasion due to non-expression of InlA and the presence of PMSC type 19 in inlA . The obtained results demonstrated that L. monocytogenes with indistinguishable PFGE profiles can be persisting or being reintroduced in beef processing facilities in the studied region and differences on their virulence potential based on their lineages and serogroups.

The establishment of norms that regulates the production and trade of Brazilian Artisanal Cheeses (BAC) has been stimulating many small farmers for this activity. The predominance of lactic acid bacteria (LAB) is a typical characteristic of BAC, which confers desirable attributes to artisanal cheeses. However, these products can be contaminated by other microbial groups, including those that indicate hygienic failures during production and may cause spoilage, or even microorganisms that pose risks to consumers’ health. A systematic review of the literature published from January 1996 to November 2020 was carried out to identify scientific data about production characteristics and microbiological aspects of BAC, with a major focus on quality and safety status of these traditional products. Studies that fulfilled the inclusion criteria indicated that artisanal chesses produced in Brazil still do not satisfactorily meet the microbiological criteria established by the national laws, mainly due to the high counts of coagulase-positive Staphylococcus and coliforms. Despite low prevalence, pathogens such as Salmonella and Listeria monocytogenes were isolated in some BAC. This review contributed to better understanding microbiological aspects of BAC, the data compiled by the authors highlight the need to improve hygiene practices along the production chain of these traditional cheeses.

RESUMO Diferentes tipos de queijos artesanais são produzidos, comercializados e consumidos no Brasil, o que impulsiona o constante desenvolvimento de normas por órgãos oficiais, como o Mapa. A criação do Suasa e do Sisbi-POA foi fundamental para esse setor, por permitir um sistema de equivalência na fiscalização e por ampliar a distribuição. Ainda, o Mapa passou a permitir que queijos artesanais produzidos com leite cru pudessem ser maturados em um período inferior a 60 dias, desde que comprovada sua inocuidade. A redução do tempo de maturação é um tema controverso e polêmico, já que não há critérios específicos que estudos científicos devem contemplar, o que permite múltiplas interpretações de dados. Com a criação e a regulamentação do selo Arte, a fiscalização dos produtos artesanais foi designada aos órgãos de agricultura, pecuária e de saúde pública, em complementação à atribuição já prevista pelo Mapa e pelo Sisbi-POA. Ainda, o selo Arte atribui aos órgãos de inspeção uma função orientadora, atividade que deveria ser prioritariamente executada por agências de extensão e associações. As normas que balizam a produção e comercialização de produtos artesanais devem ser frequentemente atualizadas, devido aos constantes avanços científicos na área e para assegurar a oferta de produtos com qualidade e inócuos aos consumidores.

Control of Salmonella spp. in food production chains is very important to ensure safe foods and minimize the risks of foodborne disease occurrence. This study aimed to identify the prevalence and main contamination sources of Salmonella spp. in a pig production chain in southern Brazil. Six lots of piglets produced at different farms were tracked until their slaughter, and samples were subjected to Salmonella spp. detection. The obtained isolates were serotyped, subjected to antimicrobial resistance testing, and pulsed field gel electrophoresis (PFGE). Salmonella spp. was detected in 160 (10.2%) samples, and not detected in pig carcasses after final washing or chilling. Among the 210 Salmonella spp. isolates, S. Typhimurium was the most prevalent (n = 101) and resistant to at least one antimicrobial. High resistance rates were detected against tetracycline (83.8%), chloramphenicol (54.3%), and trimethoprim-sulfamethoxazole (33.3%). The isolates that were non-susceptible to three or more classes of antimicrobials (n = 60) were considered multidrug-resistant (MDR), and isolates resistant to up to six of the tested antimicrobials were found. PFGE allowed the identification of genetic diversity and demonstrated that farm environment and feed supply may be sources for the dissemination of Salmonella spp. along the production chain. The results revealed the sources of Salmonella contamination in the pig production chain and highlighted the risks of antimicrobial resistance spread.

Summary: Advances in whole‐genome sequencing (WGS) technologies have documented genetic diversity and epidemiology of the major foodborne pathogen Listeria monocytogenes ( Lm ) in Europe and North America, but data concerning South America are scarce. Here, we examined the population structure and genetic diversity of this major foodborne pathogen collected in Brazil. Based on core genome multilocus sequence typing (cgMLST), isolates from lineages I ( n = 22; 63%) and II ( n = 13; 37%) were distributed into 10 different sublineages (SLs) and represented 31 new cgMLST types (CTs). The most prevalent SLs were SL9 ( n = 9; 26%), SL3 ( n = 6; 17%) and SL2 and SL218 ( n = 5; 14%). Isolates belonging to CTs L2‐SL9‐ST9‐CT4420 and L1‐SL315‐ST520‐CT4429 were collected 3 and 9 years apart, respectively, revealing long‐term persistence of Lm in Brazil. Genetic elements associated with stress survival were present in 60% of isolates (57% SSI‐1 and 3% SSI‐2). Pathogenic islands were present in 100% (LIPI‐1), 43% (LIPI‐3) and 6% (LIPI‐4) of the isolates. Mutations leading to premature stop codons were detected in the prfA and inlA virulence genes. This study is an important contribution to understanding the genomic diversity and epidemiology of Lm in South America. In addition, the results highlight the importance of using WGS to reveal Lm long‐term persistence.

Microbiological testing is an important quality management tool in the food industry. In this study, the hygiene status of beef carcasses sampled in eight Brazilian slaughterhouses was assessed by enumeration of different hygiene indicator microorganisms, and a model to establish potential associations among these counts was proposed. The carcasses (n = 464) were surface sampled at four slaughtering steps (step 1: Hide after bleeding; step 2: Carcass after hide removal; step 3: Carcass after evisceration; step 4: Carcass after end washing) and subjected to a counting of mesophilic aerobes (MA), Enterobacteriaceae (EB), total coliforms (TC), and Escherichia coli (EC) using Petrifilm™ plates. Among the sampled beef carcasses (step 4), 32 (6.9%) and 71 (15.3%) presented counts above the microbiological criteria established by (EC) No. 1441/2007 for MA and EB, respectively. Thus, indicating that improvements in slaughter hygiene and a review of process controls are demanded in some of the studied slaughterhouses. The log count differences of EC, TC, and EB from MA were considered as response variables as a function of the slaughtering steps. Differential log counts changed consistently with the steps. The measurements, including the patterns in their inherently random variability, were fairly predictable from steps 1 and 4. The results indicated that differential log counts for TC and EC are not relevant, as their concentrations and random pattern can be inferred from counts of MA and EB. The proposed model can be used as a valuable tool for the design and adoption of feasible quality control programs in beef industries. The adoption of such a tool should have a positive contribution on consumers’ health and enhance product quality.