Retail Pork Research Articles

An Investigation into the Prevalence of Clostridioides difficile in Irish Pig Abattoirs and Pork Meat Products as a Potential Source of Human Infection

Clostridioides difficile (C. difficile), once considered a predominantly nosocomial pathogen, is increasingly implicated in community-acquired infections (CA-CDIs). This study investigates the prevalence, ribotypes, and antimicrobial susceptibility of C. difficile in Irish pork products and abattoirs, with a focus on the potential public health implications. A total of 180 retail pork products and 150 pig carcase swabs from three abattoirs were examined, alongside 30 environmental lairage samples. The C. difficile isolates were characterised through ribotyping and tested in terms of antimicrobial susceptibility. No C. difficile was isolated from the retail pork, while the carcase swabs yielded a low recovery rate (0.66%). However, the lairage areas were contaminated with C. difficile (33%), and six different ribotypes were identified, including the clinically relevant RT078. The ribotypes exhibited susceptibility to the antibiotics used to treat C. difficile infection (CDI) (fidaxomicin, vancomycin, and metronidazole) but showed resistance to tetracycline (9%) and ciprofloxacin (100%). These findings align with the international findings on antimicrobial resistance in C. difficile and suggest that strict EU food safety standards could mitigate retail pork contamination risks. Nevertheless, the environmental exposure during slaughtering and handling processes presents potential transmission risks for workers.

Characterization of novel mutations involved in the development of resistance to colistin in Salmonella isolates from retail pork in Shanghai, China.

Salmonella is an important foodborne pathogen that poses a significant threat to food safety. This study aims to assess the prevalence, genomic features, and colistin-resistant mechanisms of Salmonella isolates collected from 118 retail pork samples from January 2021 to January 2022 in Shanghai, China. Overall, 46 (39.0%, 46/118) Salmonella isolates were collected, which were identified as 12 serotypes by genomic analysis, including Salmonella Typhimurium (n=17) and Salmonella London (n=6). Antimicrobial resistance profiling revealed that the resistance rate of these isolates to colistin was 13.0% (6/46), while 60.9% (28/46) exhibited multidrug-resistant. It was found that there were 51 distinct antimicrobial resistance genes in these 46 isolates, which were predominantly associated with resistance to aminoglycosides, fluoroquinolones, and β-lactams. More importantly, among six colistin-resistant isolates, two isolates (Salmonella Schwarzengrund and Salmonella Indiana) were found to carry the mcr-1 gene. The mechanism of resistance in the remaining four colistin-resistant isolates was further studied, and it was found that there were nine amino acid substitutions in PmrAB. It was demonstrated by site-directed mutagenesis that novel substitutions G53W in PmrA and I83V in PmrB led to colistin resistance in Salmonella (MIC=2 or 4μg/mL). Analysis results by real-time quantitative PCR and mass spectrometry indicated that the mutants PmrAG53W and PmrBI83V displayed higher expression levels of the gene pmrE than in the parental strain. This upregulation resulted in an increase in the production of 4-amino-4-deoxy-l-arabinose (L-Ara4N) that modified lipid A, thereby conferring resistance to colistin. These findings demonstrated that there was a high prevalence of MDR Salmonella isolates in retail pork in Shanghai, and the substitution G53W in PmrA and I83V in PmrB were independent factors contributing to the development of resistance to colistin in Salmonella via modification of lipid A with L-Ara4N.

P-1328. Contribution of Antimicrobial Resistance Data of NARMS Retail Meat in the Investigation of Persistent Strain of Salmonella Hadar Infections, Tennessee, 2021-2023

Abstract Background Reoccurring, Emerging, and Persisting (REP) strains of Salmonella have caused illness for several years and are linked to multiple sources. Some REP isolates have been detected in food products like MDR Salmonella Infantis (REPJFX01) in retail chicken. Tennessee participates in the FDA’s National Antimicrobial Resistance Monitoring System (NARMS) Retail Meat Surveillance to monitor the antimicrobial resistance of retail meat enteric bacteria isolates. We reviewed the contribution of TN NARMS data in the investigations of multistate (MS) outbreaks caused by persistent strain of S. Hadar (REPTDK01). Methods Tennessee submits every 20thSalmonella isolates from humans statewide, and all Salmonella isolates from retail meat (retail chicken, ground turkey, ground beef and pork products) purchased from 13 counties to NARMS. Sequenced isolates are uploaded to PulseNet, and resistance determinants are monitored. TN responds to CDC/FDA requests on isolates with persisting strains matching MS outbreaks. Results REPTDK01 strain was detected in 4 ground turkey (GT) isolates from TN as part of 2 multistate outbreaks in 2021 (2102MLTDK-1) and 2023 (2302MLTDK-1). Over 95% of human and non-human isolates in both outbreaks were resistant to tetracycline, streptomycin, or both, and 0.2-0.4 % to ciprofloxacin and ceftriaxone respectively. Three of the 4 GT samples were purchased from grocery stores in different counties in 2021, and two of them had the same producer establishment number. USDA issued a public health consumer alert for over 211,406 pounds of raw GT products produced by the establishment. An additional GT sample was purchased in 2023 with a different establishment number. Consumption of GT was reported by 20 (60%) of 33 cases in 2021, while no signal of GT consumption among 55 cases in 2023. TN had no human cases in both outbreaks. However, a total of 49 cases from TN were linked to another three MS outbreaks of REPTDK01 associated with backyard poultry contact between 2019 and 2023 with one death in 2022. Conclusion NARMS retail meat routine surveillance data contributes to a better understanding of source attributions of REP bacterial strains. Sharing of TN data with federal partners helped in traceback efforts, regulatory actions, and alerting the public. Disclosures All Authors: No reported disclosures

Niche-specific evolution and gene exchange of Salmonella in retail pork and chicken

Salmonella exhibits extensive genetic diversity, facilitated by horizontal gene transfer occurring within and between species, playing a pivotal role in this diversification. Nevertheless, most studies focus on clinical and farm animal isolates, and research on the pangenome dynamics of Salmonella isolates from retail stage of the animal food supply chain is limited. Here, we investigated the genomes of 950 Salmonella isolates recovered from retail chicken and pork meats in seven provinces and one municipality of China in 2018. We observed a strong correlation between Salmonella sublineage diversity and the accessory genome with meat type, revealing reduced diversity associated with increased resistance. Importantly, genes associated with antibiotic, biocide, and heavy metal resistance were unevenly distributed in Salmonella from retail chicken and pork. Pork Salmonella isolates showed a higher prevalence of copper and silver resistance genes, while chicken Salmonella isolates displayed a significant predominance of genetic determinants associated with cephalosporin and ciprofloxacin resistance. Moreover, co-occurrence patterns of resistance determinants and their interaction with mobile genetic elements also correlated with meat type. In summary, our findings shed light on how Salmonella achieves their ecological niche success driven by evolution and gene changes in the retail stage of the animal food supply chain.

Bacterial and Genetic Features of Raw Retail Pork Meat: Integrative Analysis of Antibiotic Susceptibility, Whole-Genome Sequencing, and Metagenomics.

The global antibiotic resistance crisis, driven by overuse and misuse of antibiotics, is multifaceted. This study aimed to assess the microbiological and genetic characteristics of raw retail pork meat through various methods, including the isolation, antibiotic susceptibility testing (AST), whole-genome sequencing (WGS) of selected indicator bacteria, antibiotic residue testing, and metagenomic sequencing. Samples were purchased from 10 pre-selected retail stores in Gauteng, South Africa. The samples were aseptically separated, with portions sent to an external laboratory for isolating indicator bacteria and testing for antibiotic residues. Identification of the isolated bacteria was reconfirmed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). AST was performed using the Microscan Walkaway system (Beckman Coulter, Brea, CA, USA). WGS and metagenomic sequencing were performed using the Illumina NextSeq 550 instrument (San Diego, CA, USA). The isolated E. coli and E. faecalis exhibited minimal phenotypic resistance, with WGS revealing the presence of tetracycline resistance genes. Both the isolated bacteria and meat samples harboured tetracycline resistance genes and the antibiotic residue concentrations were within acceptable limits for human consumption. In the metagenomic context, most identified bacteria were of food/meat spoilage and environmental origin. The resistome analysis primarily indicated beta-lactam, tetracycline and multidrug resistance genes. Further research is needed to understand the broader implications of these findings on environmental health and antibiotic resistance.

Collusion and Price Behavior in the U.S. Pork Industry

Abstract This research analyzes the U.S. pork industry dynamics in light of the alleged output price-fixing cartel of the largest pork processors revealed in the antitrust lawsuits filed by pork buyers beginning in 2018. The empirical evidence may suggest that pork production, the farm, wholesale, and retail values of pork, and marketing margins increased, but the farm sector share decreased in the cartel period (2009–2019), as compared with the pre-cartel period (1998–2008). The wholesale pork pricing by pork processors is consistent with oligopoly pricing in both the pre-cartel and cartel periods. While the estimated decrease in cost pass-through between these two periods may reflect an increase in the seller (oligopoly) market power of pork processors in the cartel period, this cost pass-through decrease is not statistically significant from zero. The retail pork pricing by food retailers is consistent with perfectly competitive pricing in both the pre-cartel and cartel periods.

Genome sequence, antibiotic resistance genes, and plasmids in a monophasic variant of Salmonella typhimurium isolated from retail pork.

A Salmonella isolate from retail pork was whole genome sequenced using Illumina NovaSeq6000, with a 5,320,119 bp genome and 51.06% GC content. Several antibiotic resistance genes and plasmids, including blaTEM-1, aac(6')-IIc, IncHI2, and p0111 were obtained from subsequent analysis. These findings provide vital insights into generic determinants of antimicrobial resistance (AMR) in this foodborne pathogen.

Characteristics of antimicrobial resistance in Escherichia coli isolated from retail meat products in North Carolina.

Escherichia coli is commonly used as an indicator for antimicrobial resistance (AMR) in food, animal, environment, and human surveillance systems. Our study aimed to characterize AMR in E. coli isolated from retail meat purchased from grocery stores in North Carolina, USA as part of the National Antimicrobial Resistance Monitoring System (NARMS). Retail chicken (breast, n = 96; giblets, n = 24), turkey (n = 96), and pork (n = 96) products were purchased monthly from different counties in North Carolina during 2022. Label claims on packages regarding antibiotic use were recorded at collection. E. coli was isolated from meat samples using culture-based methods and isolates were characterized for antimicrobial resistance using whole genome sequencing. Multi-locus sequence typing, phylogroups, and a single nucleotide polymorphism (SNP)-based maximum-likelihood phylogenic tree was generated. Data were analyzed statistically to determine differences between antibiotic use claims and meat type. Of 312 retail meat samples, 138 (44.2%) were positive for E. coli, with turkey (78/138; 56.5%) demonstrating the highest prevalence. Prevalence was lower in chicken (41/138; 29.7%) and pork (19/138;13.8%). Quality sequence data was available from 84.8% (117/138) of the E. coli isolates, which included 72 (61.5%) from turkey, 27 (23.1%) from chicken breast, and 18 (15.4%) from pork. Genes associated with AMR were detected in 77.8% (91/117) of the isolates and 35.9% (42/117) were defined as multidrug resistant (MDR: being resistant to ≥3 distinct classes of antimicrobials). Commonly observed AMR genes included tetB (35%), tetA (24.8%), aph(3'')-lb (24.8%), and blaTEM-1 (20.5%), the majority of which originated from turkey isolates. Antibiotics use claims had no statistical effect on MDR E. coli isolates from the different meat types (X2 = 2.21, p = 0.33). MDR was observed in isolates from meat products with labels indicating "no claims" (n = 29; 69%), "no antibiotics ever" (n = 9; 21.4%), and "organic" (n = 4; 9.5%). Thirty-four different replicon types were observed. AMR genes were carried on plasmids in 17 E. coli isolates, of which 15 (88.2%) were from turkey and two (11.8%) from chicken. Known sequence types (STs) were described for 81 E. coli isolates, with ST117 (8.5%), ST297 (5.1%), and ST58 (3.4%) being the most prevalent across retail meat types. The most prevalent phylogroups were B1 (29.1%) and A (28.2%). Five clonal patterns were detected among isolates. E. coli prevalence and the presence of AMR and MDR were highest in turkey retail meat. The lack of an association between MDR E. coli in retail meat and antibiotic use claim, including those with no indication of antimicrobial use, suggests that additional research is required to understand the origin of resistance. The presence of ST117, an emerging human pathogen, warrants further surveillance. The isolates were distinctly diverse suggesting an instability in population dynamics.

Deciphering mechanisms of bla NDM gene transmission between human and animals: a genomics study of bacterial isolates from various sources in China, 2015 to 2017.

BackgroundIn China, the bla NDM gene has been recovered from human bacterial isolates since 2011. After 2014, detections of this gene in animal and food bacterial isolates have increasingly been reported.AimWe aimed to understand how bla NDM-bearing bacteria could spread between humans, animals, and animal-derived food.MethodsA total of 288 non-duplicate Escherichia coli strains, including 130 bla NDM-carrying and 158 bla NDM-negative strains were collected from clinical (humans), food-producing animals (pigs) and food (retail pork) sources between 2015 and 2017. The strains were whole genome sequenced. Core-genome-multilocus-sequence-typing was conducted. To investigate if sequence types (STs) found in human, animal or food samples could have a prior origin in a clinical, animal or food-borne animal reservoir, discriminant analysis of principal components (DAPC) was used. Plasmids bearing bla NDM were characterised.ResultsThe 130 bla NDM-carrying E. coli strains comprised a total of 60 STs, with ST167 (10/51), ST77 (6/33) and ST48 (6/46) being most prevalent in clinical, animal and food sources, respectively. Some ST10 and ST167 strains were respectively found among all three sources sampled, suggesting they might enable transfer of bla NDM between sources. DAPC analysis indicated possible transmissions of ST167 from humans to animals and ST10 from animals to human. In 114 of 130 bla NDM-carrying isolates, bla NDM was located on an IncX3 plasmid.ConclusionThis study in a Chinese context suggests that cross-species transmission of certain STs of E. coli harbouring bla NDM on mobile elements, may facilitate the spread of carbapenem-resistant Enterobacteriaceae. Stringent monitoring of bla NDM-bearing E. coli in ecosystems is important.

Antimicrobial resistance profiles and clonal diversity of Staphylococcus epidermidis isolates from pig farms, slaughterhouses, and retail pork

Livestock-associated coagulase-negative staphylococci (CoNS), such as Staphylococcus (S.) epidermidis, have emerged as a significant reservoir of antimicrobial resistance (AMR). In the current study, the AMR profiles and genetic diversity of S. epidermidis isolates obtained from pig farms, slaughterhouses, and retail pork were analyzed. A total of 89 S. epidermidis isolates, comprising 22 methicillin-resistant (MRSE) and 67 methicillin-susceptible S. epidermidis (MSSE) isolates, were assessed to determine (i) the clonal lineages of the isolates [multilocus sequence (MLST), agr, and staphylococcal cassette chromosome mec (SCCmec) types], (ii) the profiles of AMR phenotypes, and (iii) the carriage of genetic factors associated with major AMR phenotypes and zinc chloride resistance. Two dominant clonal lineages of S. epidermidis, ST100 and ST570, were observed on pig farms, especially in healthy pigs. In addition, potential transmission of pig-associated ST100 MRSE-SCCmec V and ST100 MSSE to farm workers was identified. The high prevalence of ST100 and ST570 isolates in pig farms was associated with high levels of AMR and zinc chloride resistance. In relation to resistance phenotypes, higher carriage rates of resistance genes, such as β–lactams (mecA), phenicols (fexA), and tetracyclines [tet(K), tet(L), tet(S), tet(M), and tet(O)], were identified in pig farm-associated isolates. Furthermore, cfr-mediated linezolid resistance was detected in 14 MSSE isolates from pig farms. This study is the first to provide important insights into the clonal structures and AMR profiles of S. epidermidis isolates collected from healthy pigs, carcass/pork samples, and human workers in Korea.

Prevalence and Virulence Determinants of Staphylococcus aureus in Wholesale and Retail Pork in Wuhan, Central China.

Staphylococcus aureus is one of the major foodborne pathogens and can cause serious foodborne illness in humans by foods contaminated with S. aureus enterotoxins. In recent years, livestock-associated S. aureus has been a major public health concern for humans and has emerged in various countries globally. China is one of the largest producers of pigs and pork in the world. However, there are few studies on the detailed genotypic and pathogenic characterization of pork-associated S. aureus in China. In this study, the prevalence, antimicrobial resistance, and genotypic characteristics of S. aureus in raw pork in Wuhan, China, were investigated through multilocus sequence typing (MLST), staphylococcal protein A gene (spa) typing, and whole-genome sequencing analysis. A total of 518 S. aureus isolates (16.9%) were isolated from 3067 retail and wholesale pork samples. The prevalence of S. aureus in retail pork (22.7%) was significantly higher than in wholesale pork (15.1%), while the proportion of multidrug-resistant (MDR) isolates in wholesale pork (12.9%) was significantly higher than in retail pork (6.2%). Among the isolates, 10.8% were resistant to three or more antibiotics, with higher rates of resistance to penicillin (88.8%) and erythromycin (58.1%). A total of 28 sequence types (STs) were identified in the 518 isolates, and the predominant type was ST7 (57.5%), followed by ST5 (9.1%). In addition, based on the whole-genome sequences of 39 representative strains, 17 spa types were identified among the isolates, of which t899, t091, and t437 were the most common. Furthermore, 19 staphylococcal enterotoxin (SE) and SE-like (SEl) toxin genes were detected in the isolates, of which selw was the most common type (100%), followed by sei, sem, seo, seu, and selv (46.2%); sey (35.9%); and sea, seg, and sen (33.3%). This study found for the first time that ST7-t091-selw and ST9-t899-SCCmecXII-selw were the predominant genotypes of S. aureus in pork in China, which indicated the spreading of S. aureus with multiple virulence factors, especially with new SE/SEl types in pigs and pork, is a serious new challenge for food safety. Good hygiene and good production practices to prevent interspecies transmission and cross-contamination of S. aureus in the pig-pork chain are of great significance to public health.

Quantitative Risk Assessment of Susceptible and Ciprofloxacin-Resistant Salmonella from Retail Pork in Chiang Mai Province in Northern Thailand.

The adverse human health effects as a result of antimicrobial resistance have been recognized worldwide. Salmonella is a leading cause of foodborne illnesses while antimicrobial resistant (AMR) Salmonella has been isolated from foods of animal origin. The quantitative risk assessment (RA) as part of the guidelines for the risk analysis of foodborne antimicrobial resistance was issued by the Codex Alimentarius Commission more than a decade ago. However, only two risk assessments reported the human health effects of AMR Salmonella in dry-cured pork sausage and pork mince. Therefore, the objective of this study was to quantitatively evaluate the adverse health effects attributable to consuming retail pork contaminated with Salmonella using risk assessment models. The sampling frame covered pork at the fresh market (n = 100) and modern trade where pork is refrigerated (n = 50) in Chiang Mai province in northern Thailand. The predictive microbiology models were used in the steps where data were lacking. Susceptible and quinolone-resistant (QR) Salmonella were determined by antimicrobial susceptibility testing and the presence of AMR genes. The probability of mortality conditional to foodborne illness by susceptible Salmonella was modeled as the hazard characterization of susceptible and QR Salmonella. For QR Salmonella, the probabilistic prevalences from the fresh market and modern trade were 28.4 and 1.9%, respectively; the mean concentrations from the fresh market and modern trade were 346 and 0.02 colony forming units/g, respectively. The probability of illness (PI) and probability of mortality given illness (PMI) from QR Salmonella-contaminated pork at retails in Chiang Mai province were in the range of 2.2 × 10−8–3.1 × 10−4 and 3.9 × 10−10–5.4 × 10−6, respectively, while those from susceptible Salmonella contaminated-pork at retails were in the range 1.8 × 10−4–3.2 × 10−4 and 2.3 × 10−7–4.2 × 10−7, respectively. After 1000 iterations of Monte Carlo simulations of the risk assessment models, the annual mortality rates for QR salmonellosis simulated by the risk assessment models were in the range of 0–32, which is in line with the AMR adverse health effects previously reported. Therefore, the risk assessment models used in both exposure assessment and hazard characterization were applicable to evaluate the adverse health effects of AMR Salmonella spp. in Thailand.

Prevalence, Serotypes, and Antimicrobial Resistance Patterns of Non-Typhoid Salmonella in Food in Northern Taiwan.

Salmonella is one of the most common bacteria causing food poisoning worldwide. We evaluated the prevalence, the serotypes, and the antimicrobial resistance (AMR) of Salmonella isolates from many kinds of food, particularly pork and chicken in retail, in Taiwan between January 2017 and December 2019. The E-test was used to assess antimicrobial susceptibility and a polymerase chain reaction was performed for serotyping. A total of 459 different foods were investigated, and 117 Salmonella strains were isolated. Retail pork and chicken were the most common Salmonella-contaminated foods (64.1% and 29.1%, respectively). Of the 117 isolates, 23 serotypes were identified. The serotypes Derby (16.2%), Anatum (13.7%), and Agona (8.5%) were the most prevalent. The resistance rates to ciprofloxacin, ceftriaxone, and carbapenem were 41.9%, 11.1%, and 1.7%, respectively. The Derby and Anatum serotypes were prevalent in chicken and pork; the Anatum serotype had significantly higher ciprofloxacin and ceftriaxone resistance rates and was highly prevalent in 2017 and 2018. Multi-locus sequence typing analysis revealed that the 58 randomly chosen Salmonella isolates belonged to 18 sequence types (STs). ST64 (Anatum, 16 out of 58, 27.6%) was the most common, followed by ST321 (Muenster, 6/58, 10.3%), ST831 (Give, 5/58, 8.6%), ST155 (London, 4/58, 6.9%) and ST314 (Kentucky, 4/58, 6.9%). Multidrug-resistant Salmonella strains were remarkably observed in the serotypes Anatum (ST64) and Goldcoast (ST358). This study revealed that retail pork was commonly contaminated with antimicrobial-resistant Salmonella. Thus, periodic investigations of Salmonella serotypes and AMR are needed.

Characterization of blaNDM-positive Enterobacteriaceae reveals the clonal dissemination of Enterobacter hormaechei coharboring blaNDM and tet(X4) along the pork production chain

The convergence of blaNDM and tet(X4) in a single strain has been sporadically detected in diverse pathogens from different sources, causing widespread concerns. However, the genomic characterization of blaNDM-positive or blaNDM- and tet(X4)-coharboring Enterobacteriaceae in the pork production chain, as well as how they emerged, remains mostly unclear. In this study, forty blaNDM-positive Enterobacteriaceae, including eleven tet(X4)- and blaNDM-coharboring isolates, were isolated from a pig slaughterhouse and a retail pork market in Yangzhou, China. This is the first report presenting the coexistence of blaNDM and tet(X4) in Enterobacter hormaechei, and it was demonstrated that they could spread clonally from the slaughterhouse to the retail market. Furthermore, we also discovered that the slaughterhouse served as a possible reservoir for blaNDM-positive ST48 E. coli strains. The blaNDM-bearing IncX3 plasmid was found in twenty blaNDM-positive strains as well as all blaNDM-5- and tet(X4)-coharboring strains, and most of them (30/31) were conjugative. In Klebsiella aerogenes NTT31XS, blaNDM was found on a nonconjugative ~17 kb IncX3 plasmid. When compared to the typical IncX3 plasmid, it deleted large-scale regions, which might imply a low fitness cost dissemination strategy for the IncX3 plasmid. Despite being from different species, tet(X4)-bearing plasmids carried by all blaNDM-5- and tet(X4)-coharboring strains were nonconjugative and had identical architectures. Phylogenetic analysis suggested that the formation of K. aerogenes NTT31XS may have been mediated by translocation of tet(X4)-positive circular intermediates and recombination events of regions in typical blaNDM-bearing IncX3 plasmids. Overall, this study expounded the prevalence and transmission characteristics of blaNDM-positive Enterobacteriaceae strains in the two links of pork production chain before they were exposed to humans, and expanded our knowledge of the molecular features and evolutionary history of tet(X4)- and blaNDM-5-coharboring strains. Strict control measures should be implemented to prevent the spread of such pathogens along food production chains.

Analysis of Antimicrobial Resistance in Non-typhoidal Salmonella Collected From Pork Retail Outlets and Slaughterhouses in Vietnam Using Whole Genome Sequencing.

Non-typhoidal salmonella (TS) remains a significant health burden worldwide. In Vietnam, pork accounts for 70% of the total meat consumed, and contamination with Salmonella is high. High levels of antimicrobial resistance (AMR) have emerged among porcine NTS and of particular concern is the emergence of colistin resistance, a “last defense” antibioic against multi-drug resistant (MDR) Gram-negative pathogens. This study aimed to investigate the antibiotic susceptibility of 69 NTS isolates collected from the pork retail outlets and slaughterhouses in Vietnam during 2014 a nd 2018/19. Phenotypic testing and whole genome sequencing was used to assess the serotype and AMR gene profiles of the 69 NTS isolates. Seventeen different serotypes were identified, of which S. enterica subsp enterica serotype Typhimurium was the most common followed by S. ser. Rissen, S. ser. London, S. ser. Anatum, and S. ser. Derby. Phenotype AMR was common with 41 (59.4%) isolates deemed MDR. MDR strains were most common in slaughterhouses (83%) and supermarkets (75%) and lowest in traditional markets (38%) and convenience stores (40%). Colistin resistance was identified in 18 strains (15 resistant, three intermediate) with mcr-1 identified in seven isolates (S. ser. Meleagridis, S. Rissen, S. Derby) and mcr-3 in two isolates (S. Typhimurium). This includes the first mcr positive S. Meleagridis to our knowledge. Surprisingly, boutique stores had high levels (60%) of MDR isolates including 5/20 isolates with mcr-1. This study demonstrates that pork from modern retail stores classed as supermarkets or boutique (with pork claiming to be high quality, traceable, environmentally friendly marketed toward higher income consumers) still contained NTS with high levels of AMR.

Profiles of Non-aureus Staphylococci in Retail Pork and Slaughterhouse Carcasses: Prevalence, Antimicrobial Resistance, and Genetic Determinant of Fusidic Acid Resistance.

As commensal colonizers in livestock, there has been little attention on staphylococci, especially non-aureus staphylococci (NAS), contaminating meat production chain. To assess prevalence of staphylococci in retail pork and slaughterhouse carcass samples in Korea, we collected 578 samples from Korean slaughterhouses (n=311) and retail markets (n=267) for isolation of staphylococci and determined antimicrobial resistance phenotypes in all the isolates. The presence of and prevalence of fusB-family genes (fusB, fusC, fusD, and fusF) and mutations in fusA genes were examined in fusidic acid resistant isolates. A total of 47 staphylococcal isolates of 4 different species (Staphylococcus aureus, n=4; S. hyicus, n=1; S. epidermidis, n=10; Mammaliicoccus sciuri, n=32) were isolated. Fusidic acid resistance were confirmed in 9/10 S. epidermidis and all of the 32 M. sciuri (previously S. sciuri) isolates. Acquired fusidic acid resistance genes were detected in all the resistant strains; fusB and fusC in S. epidermidis and fusB/C in M. sciuri. Multi-locus sequence type analysis revealed that ST63 (n=10, 31%) and ST30 (n=8, 25%) genotypes were most prevalent among fusidic acid resistant M. sciuri isolates. In conclusion, the high prevalence of fusB-family genes in S. epidermidis and M. sciuri strains isolated from pork indicated that NAS might act as a reservoir for fusidic acid resistance gene transmissions in pork production chains.

Tracing enterococci persistence along a pork production chain from feed to food in China.

The prevalence and transmission of vancomycin-resistant Enterococcus (VRE) in enterococci being as probiotics has been neglected in the scientific literature. The application of enterococci in feed, food and health products may cause VRE transmission through the food chain. This study evaluated phenotypic resistance of Enterococcus species to 20 antibiotics along a pork production chain from feed to food. It also assessed the genetic diversity of Enterococcus faecium isolates. A total of 510 samples (feed, n = 70; swine manure, n = 400; swine carcasses, n = 20, and retail pork, n = 20) were collected in Beijing, China. A total of 328 enterococci isolates with 275 E. faecium and 53 Enterococcus faecalis were identified using 16 S rRNA. Antimicrobial susceptibility to all enterococci isolates was conducted using the K–B method for 20 antibiotics from 9 categories. Multilocus sequence typing (MLST) was conducted on the E. faecium isolates to survey the dissemination of enterococci in the pig industry. The results showed that only 26 enterococci isolates were sensitive to the 20 antibiotics, while half of the isolates (164/328) had acquired multi-drug resistance. The resistant rate to furazolidone was 68.60%, followed by 42.99% to tetracycline. One vancomycin-resistant E. faecium isolates were isolated from feed origin and 2 from manure origin, with minimum inhibitory concentrations to vancomycin of 1,024, 64, and 64 μg/mL, respectively. The MLST outcomes showed that the 275 E. faecium isolates belonged to 11 sequence types (ST) including ST40, ST60, ST94, ST160, ST178, ST296, ST361, ST695, ST726, ST812 and ST1014. The ST of the feed-sourced VRE was ST1014, while the 2 manure-sourced VRE was ST69. ST1014 evolved from ST78, which was the dominant clonal complex in most cities of China, leading to the spreading of VRE. These findings revealed the potential safety hazards of commercial probiotic enterococci in China and showed that there is a risk of the VRE horizontally transferring from feed to food.

Prevalence and characterization of -methicillin-resistant Macrococcus spp. in food producing animals and meat in Switzerland in 2019.

The prevalence of methicillin-resistant Macrococcus spp. in calves and pigs at slaughterhouses and in retail beef and pork meat was determined using samples taken in 2019 within the framework of the national monitoring of methicillin-resistant Staphylococcus aureus in food producing animals in Switzerland. The isolates were submitted to antimicrobial susceptibility testing of 19 antibiotics and to molecular techniques (e.g. PCR, microarray, WGS) for the identification of resistance genes, elements containing the methicillin resistance genes mec and sequence type (ST). Methicillin-resistant Macrococcus spp. (M. caseolyticus (n=38), M. bohemicus (n=4) and Macrococcus spp. (n=2)) were isolated in 40 of 299 nasal swabs from calves representing a prevalence of 13,38 % (95 % CI, 9,98 % - 17,70 %), and in four of 303 nasal swabs from pigs [1,32 % (95 % CI, 0,36 % - 3,35 %)]. One of 311 samples of Swiss pork meat contained a Macrococcus sp. [0,32 % (95 % CI, 0,01 % - 1,78 %)], and four of 309 beef meat samples (260 domestic and 49 imported) contained M. caseolyticus [1,29 % (95 % CI, 0,35 % - 3,28 %)]. The M. caseolyticus strains belonged to diverse STs, with ST21 being the most common in both pigs and calves. The mecD gene was located on Macrococcus resistance island mecD (McRImecD) in 42 strains and on staphylococcal cassette chromosome mec (SCCmecD) in three strains, while mecB was found on plasmids in four strains. In addition to resistance to β-lactams, the strains also exhibited resistance to tetracycline (n=17; tet(L), tet(K), tet(M)), streptomycin (n=13; str, ant(6)-Ia, rpsL mutation [K56R in ribosomal protein S12]), kanamycin (n=10; aac(6')-Ie - aph(2'')-Ia, aph(2')-Ib, aph(2')-Ic, ant(4')-Ia), clindamycin (n=9; erm(B), erm(45)), erythromycin (n=9; erm(B), msr(G), erm(45)), fusidic acid (n=9; fusC) and gentamicin (n=1; aac(6')-Ie - aph(2'')-Ia). This study represents the first national prevalence study of methicillin-resistant Macrococcus spp. in pigs, calves, pork and beef meat in Switzerland and revealed a reservoir of genetically diverse strains carrying several resistance traits.

Whole-Genome Epidemiology and Characterization of Methicillin-Susceptible Staphylococcus aureus ST398 From Retail Pork and Bulk Tank Milk in Shandong, China.

Staphylococcus aureus (S. aureus) is now regarded as a zoonotic agent. Methicillin-susceptible S. aureus (MSSA) ST398 is a livestock-associated bacterium that is most prevalent in China, but there are currently no data available for Shandong. Therefore, the aim of this study was to investigate the epidemiology and characterization of MSSA ST398 from retail pork and bulk tank milk (BTM) in Shandong. A total of 67 S. aureus isolates were collected from retail pork between November 2017 and June 2018. Among the isolates, high antimicrobial resistance rates were observed for penicillin (97.0%), and 92.5% of the isolates were multi-drug resistant (MDR). Eight sequence types (STs) were identified in the retail pork isolates, and the predominant type was ST15 (n=26), which was followed by ST398 (n=14). Staphylococcal protein A gene (spa) typing identified spa types t034 and t1255 in MSSA ST398 from retail pork. Using whole-genome sequencing analysis, we described the phylogeny of 29 MSSA ST398 isolates that were obtained from retail pork (n=14) and BTM (n=15). The phylogenetic tree showed that the MSSA ST398 isolates from different sources had the same lineage. Among the 29 MSSA ST398 isolates, five resistance genes were detected, and all isolates carried DHA-1. Fifteen toxin genes were detected, and all isolates carried eta, hla, and hlb. In conclusion, this study found that a high risk for MSSA ST398 was present in retail pork and BTM. These findings have major implications for how investigations of MSSA ST398 outbreaks should be conducted in the One-Health context.

Shiga Toxin–Producing Escherichia coli (STEC) and STEC-Associated Virulence Genes in Raw Ground Pork in Canada

Shiga toxin-producing Escherichia coli (STEC) O157:H7/nonmotile and some non-O157 STEC strains are foodborne pathogens. In response to pork-associated O157 STEC outbreaks in Canada, we investigated the occurrence of STEC in Canadian retail raw ground pork during the period of 1 November 2014 to 31 March 2016. Isolated STEC strains were characterized to determine the Shiga toxin gene (stx) subtype and the presence of virulence genes encoding intimin (eae) and enterohemorrhagic E. coli hemolysin (hlyA). O157 STEC and non-O157 STEC strains were isolated from 1 (0.11%) of 879 and 13 (2.24%) of 580 pork samples, respectively. STEC virulence gene profiles containing both eae and hlyA were found only in the O157 STEC (stx2a, eae, hlyA) isolate. The eae gene was absent from all non-O157 STEC isolates. Of the 13 non-O157 STEC isolates, two virulence genes of stx1a and hlyA were found in four (30.8%) O91:H14 STEC isolates, whereas one virulence gene of stx2e, stx1a, and stx2a was identified in five (38.5%), two (15.4%), and one (7.7%) STEC isolates, respectively, of various serotypes. The remaining non-O157 STEC isolate carried stx2, but the subtype is unknown because this isolate could not be recovered for sequencing. O91:H14 STEC (stx1a, hlyA) was previously reported in association with diarrheal illnesses, whereas the other non-O157 STEC isolates identified in this study are not known to be associated with severe human illnesses. Virulence gene profiles identified in this study indicate that the occurrence of non-O157 STEC capable of causing severe human illness is rare in Canadian retail pork. However, O157 STEC in ground pork can occasionally occur; therefore, education regarding the potential risks associated with STEC contamination of pork would be beneficial for the public and those in the food industry to help reduce foodborne illnesses.