Comparative genomics and antibiotic resistance of Yersinia enterocolitica obtained from a pork production chain and human clinical cases in Brazil.

Previous work found a high similarity of macro-restriction patterns for isolates of Yersinia enterocolitica 4/O:3 obtained at a pork production chain from Minas Gerais, Brazil. Herein we aimed to determine the clonality and the antibiotic resistance profiles of a subset of these isolates (n = 23) and human clinical isolates (n = 3). Analysis based on whole genome sequencing (WGS) showed that the isolates were distributed into two major clades based on single nucleotide polymorphisms (SNP) with one isolate defining Clade A (isolate R31) and remaining isolates (n = 25, 96.2%) defining Clade B. Seven clonal groups were identified. The inclusion of isolate R31 as a distinct clonal group was due to the presence of several phage-related genes, allowing its characterization as serotype O:5 by WGS. Disk-diffusion assays (14 antibiotics) identified 13 multidrug resistant isolates (50.0%). Subsequent sequence analysis identified 17 different antibiotic resistance related genes. All isolates harbored blaA (y56 beta-lactamase), vatF, rosA, rosB and crp, while nine isolates harbored a high diversity of antibiotic resistance related genes (n = 13). The close genetic relationship among Y. enterocolitica obtained from a pork production chain and human clinical isolates in Brazil was confirmed, and we can highlight the role of swine in the potential transmission of an antibiotic-resistant clones of a pathogenic bio-serotype to humans, or the transmission of these resistant bacteria from people to animals. The role of veterinary antibiotic use in this process is unclear.

Pathogenic potential and antibiotic resistance of Yersinia enterocolitica, a foodborne pathogen limited to swine tonsils in a pork production chain from Southern Brazil.

In this study, we aimed to characterize the distribution of Yersinia enterocolitica in a pork production chain in Brazil, as well as the virulence profile and antibiotic resistance of the obtained isolates. Samples from 10 pig lots obtained from finishing farms (water, feed, and barn floors, n = 30), slaughterhouse (lairage floors, carcasses at four processing steps, tonsils, and mesenteric lymph nodes, n = 610), and processing (end cuts, processing environment, n = 160) were obtained in Paraná state, Brazil, and subjected to Y. enterocolitica detection by ISO 10,273. The obtained isolates were identified based on biochemical and molecular features (16 s rRNA, inv, bioserotyping) and subjected to PCR assays to detect virulence (ail, ystA, ystB, virF, myfA, fepA, fepD, fes, tccC, ymoA, hreP, and sat) and multidrug resistance-related genes (emrD, yfhD, and marC). Also, isolates were subjected to disk diffusion test to characterize their resistance against 17 antibiotics from 11 classes and to pulsed field gel electrophoresis (PFGE) after XbaI macro-restriction. Y. enterocolitica was detected in a single sample (tonsil), and the obtained three isolates were characterized as serotype O:3, harboring ail, ystA, virF, myfA, tccC, ymoA, hreP, emrD, yfhD, and marC, and resistant to all tested antibiotics. The three isolates presented identical macro-restriction profiles by PFGE, also identical to isolates obtained from Minas Gerais, other Brazilian state; one selected isolate was identified as biotype 4. Despite the low occurrence of Y. enterocolitica in the studied pork production, the virulence potential and the antibiotic resistance profiles of the isolates demonstrated their pathogenic potential, and the macro-restriction profiles indicate strains descending from a common subtype in the pork production chain of two Brazilian States.

Persistence of Yersinia enterocolitica bio-serotype 4/O:3 in a pork production chain in Minas Gerais, Brazil.

Yersinia enterocolitica bio-serotype 4/O:3 was previously identified in a pork production chain in Brazil and the obtained isolates presented high identity by pulsed-field gel electrophoresis (PFGE, XbaI). For the current study, an additional 147 porcine samples (tonsils = 100, palate = 30, head meat = 17) were collected from the same pork production chain 2-years later and 14 (9.5%) tested positive for Y. enterocolitica. Isolates (n = 24, 1 to 2 per positive sample) were bio-serotype 4/O:3 and harbored virulence genes ail, inv, wbbU, virF, myfA, ystA, ymoA, hreP and sat, and the multidrug resistance related genes emrD, marC and yfhD. PFGE (XbaI) demonstrated no differences among isolates (100% similarity) and were identical to some Y. enterocolitica isolates (n = 13) obtained previously from the same pork chain. A second PFGE analysis (NotI) confirmed the high degree of similarity among isolates obtained over time, demonstrating the persistence of an apparent clonal Y. enterocolitica bio-serotype 4/O:3 in this particular pork production chain in Brazil.

Multidrug Resistance and Virulence Profiles of Salmonella Isolated from Swine Lymph Nodes.

Salmonella spp. is a foodborne pathogen present in the pork production chain, leading to potential contamination of end products and causing salmonellosis cases and outbreaks worldwide. The emergence of multidrug-resistant (MDR) Salmonella spp., especially isolates obtained from animal origin food, is a global concern. This study aimed to isolate Salmonella from swine mesenteric lymph nodes (MLN) and to characterize the virulence and antibiotic resistance profiles. MLN samples were obtained from a swine slaughterhouse and subjected to Salmonella spp. isolation. Ten MLN samples were positive and 29 isolates were identified based on PCR (invA and ompC) and serotyping: Derby, Cerro, and Give. Pulsed-field gel electrophoresis allowed to group the isolates based on their serotypes, resulting in three major clusters. All isolates presented the virulence-related genes pefA, sipA, sopB, spaN, and pagC. Relatively high numbers of Salmonella spp. were resistant to neomycin, polymyxin B, ciprofloxacin, tetracycline, and nalidixic acid. Furthermore, 25 isolates presented simultaneous resistance to three or more antibiotic classes, being characterized as MDR. The obtained results confirmed the relevance of swine as reservoirs of Salmonella spp. in the pork production chain and demonstrated the MDR profiles of isolates. Proper control and surveillance are required to avoid the contamination of end products.

Occurrence, quantification, pulse types, and antimicrobial susceptibility of Salmonella sp. isolated from chicken meat in the state of Paraná, Brazil.

The aim of this work was to verify the occurrence, quantification, pulse types, and antimicrobial susceptibility profiles of Salmonella sp. isolated from chicken meat produced and marketed in the state of Paraná, considered to be the state with the highest production of poultry meat in Brazil. Ninety-five of 300 (31.5%) frozen cuts of chicken were found to contain Salmonella sp., and 98 different isolates of Salmonella sp. were cultured from the positive samples. Quantification showed low Salmonella sp. loading, ranging from 0.12 to 6.4 MPN/g. The antimicrobial resistance test was performed against 16 agents from 6 different classes. All isolates were sensitive to meropenem, imipenem, chloramphenicol, and amikacin. The highest resistance rates were observed for nalidixic acid (95%), tetracycline (94%), doxycycline (94%), ampicillin (87%), amoxicillin with clavulanic acid (84%), ceftriaxone (79%), and ciprofloxacin (76%). A total of 84 (85.7%) of the isolates were identified with a multidrug resistant profile, 13 of which were found to have encoding genes extended-spectrum beta-lactamase (ESBL), especially blaCTX-M-2 e blaTEM-1. The major serovars identified were S. Typhimurium (43%) and S. Heidelberg (39%). The third most isolated serovar was S. Ndolo (6%), without previous reports of its presence in poultry meat in Brazil. Molecular characterization of S. Typhimurium and S. Heidelberg isolates by pulsed field gel electrophoresis (PFGE) showed a clonal relationship between all isolates of the same serovar (genetic similarity greater than 80%). Isolates of S. Typhimurium and S. Heidelberg with 100% similarity were found in up to five different geographic regions of the state, showing the potential for the spread of this pathogen in the Paraná poultry chain. Epidemiological surveys like this are important to understand the dynamics of dissemination and to monitor the prevalence of pathogens in the final products of poultry chains. In addition, to know the resistance profile of strains of Salmonella sp. present in food that contributes to the adoption of faster and more effective therapeutic measures, when necessary.

Yersinia enterocolitica in a Brazilian pork production chain: Tracking of contamination routes, virulence and antimicrobial resistance.

This study aimed to track Yersinia enterocolitica contamination in a pork production chain in Minas Gerais, Brazil, and to characterize the virulence and antibiotic resistance of isolates. Samples were collected from four different steps of the pork production chain (pig farm, carcass, processing environment and end product; n = 870), and tested for the presence of Y. enterocolitica. The pathogen was detected in 8 samples (palatine tonsils = 5; mesenteric lymph nodes = 2; carcass after bleeding = 1), from which 16 isolates were obtained and identified as Y. enterocolitica bioserotype 4/O:3. XbaI macrorestriction allowed the clustering of isolates in 5 pulsetypes, and the identification of identical profiles of Y. enterocolitca isolated from different samples. All isolates were positive for the virulence related genes ail, virF, myfA, ystA, tccC, ymoA, hreP and sat, and negative for ystB, ystC, fepA, fepD and fes. Considering 17 antibiotics from 11 classes, only ciprofloxacin and kanamycin were effective against all isolates, and three multidrug resistance profiles were identified among them, with simultaneous resistance to 9 of 11 classes. All isolates presented positive results for emrD, yfhD and marC, related to multidrug resistance. The results of this study demonstrated the contamination routes of Y. enterocolitica within the assessed pork production chain, and highlighted the pathogenic potential and antibiotic resistance of this foodborne pathogen.