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Foodborne illness is exacerbated by novel and emerging pathotypes, persistent contamination, antimicrobial resistance, an ever-changing environment, and the complexity of food production systems. Sporadic and outbreak events of common foodborne pathogens like Shiga toxigenic E. coli (STEC), Salmonella, Campylobacter, and Listeria monocytogenes are increasingly identified. Methods of controlling human infections linked with food products are essential to improve food safety and public health and to avoid economic losses associated with contaminated food product recalls and litigations. Bacteriophages (phages) are an attractive additional weapon in the ongoing search for preventative measures to improve food safety and public health. However, like all other antimicrobial interventions that are being employed in food production systems, phages are not a panacea to all food safety challenges. Therefore, while phage-based biocontrol can be promising in combating foodborne pathogens, their antibacterial spectrum is generally narrower than most antibiotics. The emergence of phage-insensitive single-cell variants and the formulation of effective cocktails are some of the challenges faced by phage-based biocontrol methods. This review examines phage-based applications at critical control points in food production systems with an emphasis on when and where they can be successfully applied at production and processing levels. Shortcomings associated with phage-based control measures are outlined together with strategies that can be applied to improve phage utility for current and future applications in food safety.

Salmonella Enteritidis (SE) remains a leading cause of human illness worldwide, and its persistence in poultry environments might be partially attributed to their ability to form biofilm. This study compared the biofilm capacity of 15 SE and 24 Salmonella Kentucky (SK) isolates from poultry products and processing facilities to uncover genetic factors driving biofilm heterogeneity. Biofilm formation and curli/cellulose production were evaluated at 20–22 °C. Genomic analyses included phylogenetic reconstruction, comparative system profiling, SNP variation, and BLASTp v2.17.0 comparisons. Phenotypic assays showed that most SE isolates (73%) were strong biofilm formers, while the majority of SK isolates (62%) failed to form biofilms, despite many carrying the complete curli–cellulose gene set and other biofilm-associated genes. Genomic analysis identified 124 biofilm-related genes, 108 of which were conserved across all isolates, and revealed 24 variants with potential functional impact. Mutations in cellulose biosynthesis (bcs) genes were linked to weaker biofilms, whereas nonsynonymous variants in tol family genes may impair flagellar biosynthesis and matrix stability. These findings demonstrate that genetic variation, not just gene presence, shapes biofilm phenotypes and highlight key molecular targets that may explain why SE persists in poultry production while SK is less successful.

Recent concerns over linkages between antimicrobial resistance in human pathogens and antimicrobial use in livestock have prompted researchers to investigate management strategies that reduce the current reliance on in-feed tylosin to control liver abscesses in feedlot cattle. A total of 7,576 crossbred yearlings were allocated to the study (~253 animals/pen, 10 replicate pens per treatment) and individually randomized to one of three treatments. Tylosin phosphate (11 ppm) was included in-feed (1) for the first 125 days on feed (DOF) ( ), (2) for DOF 41 to 161 ( ), or (3) for the entire feeding period ( ; day 0-161). Fecal composites were collected from the pen floor on days 0, 81, and 160 of the finishing period. Serial dilutions were spread plated for enumeration of enterococci on Bile Esculin Azide (BEA) agar and BEA amended with 8 μg/ml erythromycin. Results indicated that although the proportion of Ery enterococci increased with DOF ( < 0.01), neither treatment ( = 0.34) or treatment × DOF ( = 0.37) affected antimicrobial resistance. Of the 538 isolates, 97% were enterococci, with mixed species isolated early in the feeding period and only isolated at the end. Isolates were most frequently resistant to tylosin (86%), erythromycin (84%), and doxycycline (31%). Macrolide and tetracycline resistant isolates harbored (B), C, and (L), (M), (O) genes, respectively. Overall, the proportion of Ery enterococci increased ( < 0.05) in all three treatments over the feeding period. Compared to the control cattle, cattle had more severe ( < 0.05) liver abscesses, while there was a trend ( < 0.08) for this response in cattle. There was no difference ( > 0.05) in total liver abscesses, growth performance, carcass traits, morbidity, or mortality among treatments. These results support the potential to reduce the duration and therefore quantity of tylosin administered to feedlot cattle during the feeding period without impacting animal productivity.

This study aimed to compare antimicrobial resistance (AMR) in extended-spectrum cephalosporin-resistant and generic Escherichia coli from a One Health continuum of the beef production system in Alberta, Canada. A total of 705 extended-spectrum cephalosporin-resistant E. coli (ESCr) were obtained from: cattle feces (CFeces, n = 382), catch basins (CBasins, n = 137), surrounding streams (SStreams, n = 59), beef processing plants (BProcessing, n = 4), municipal sewage (MSewage; n = 98) and human clinical specimens (CHumans, n = 25). Generic isolates (663) included: CFeces (n = 142), CBasins (n = 185), SStreams (n = 81), BProcessing (n = 159) and MSewage (n = 96). All isolates were screened for antimicrobial susceptibility to 9 antimicrobials and two clavulanic acid combinations. In ESCr, oxytetracycline (87.7%), ampicillin (84.4%) and streptomycin (73.8%) resistance phenotypes were the most common, with source influencing AMR prevalence (p < 0.001). In generic E. coli, oxytetracycline (51.1%), streptomycin (22.6%), ampicillin (22.5%) and sulfisoxazole (14.3%) resistance were most common. Overall, 88.8% of ESCr, and 26.7% of generic isolates exhibited multi-drug resistance (MDR). MDR in ESCr was high from all sources: CFeces (97.1%), MSewage (96.9%), CHumans (96%), BProcessing (100%), CBasins (70.5%) and SStreams (61.4%). MDR in generic E. coli was lower with CFeces (45.1%), CBasins (34.6%), SStreams (23.5%), MSewage (13.6%) and BProcessing (10.7%). ESBL phenotypes were confirmed in 24.7% (n = 174) ESCr and 0.6% of generic E. coli. Prevalence of bla genes in ESCr were blaCTXM (30.1%), blaCTXM-1 (21.6%), blaTEM (20%), blaCTXM-9 (7.9%), blaOXA (3.0%), blaCTXM-2 (6.4%), blaSHV (1.4%) and AmpC β-lactamase blaCMY (81.3%). The lower AMR in ESCr from SStreams and BProcessing and higher AMR in CHumans and CFeces likely reflects antimicrobial use in these environments. Although MDR levels were higher in ESCr as compared to generic E. coli, AMR to the same antimicrobials ranked high in both ESCr and generic E. coli sub-populations. This suggests that both sub-populations reflect similar AMR trends and are equally useful for AMR surveillance. Considering that MDR ESCr MSewage isolates were obtained without enrichment, while those from CFeces were obtained with enrichment, MSewage may serve as a hot spot for MDR emergence and dissemination.

Antimicrobial resistance (AMR) has important implications for the continued use of antibiotics to control infectious diseases in both beef cattle and humans. AMR along the One Health continuum of the beef production system is largely unknown. Here, whole genomes of presumptive extended-spectrum β-lactamase E. coli (ESBL-EC) from cattle feces (n = 40), feedlot catch basins (n = 42), surrounding streams (n = 21), a beef processing plant (n = 4), municipal sewage (n = 30), and clinical patients (n = 25) are described. ESBL-EC were isolated from ceftriaxone selective plates and subcultured on ampicillin selective plates. Agreement of genotype-phenotype prediction of AMR ranged from 93.2% for ampicillin to 100% for neomycin, trimethoprim/sulfamethoxazole, and enrofloxacin resistance. Overall, β-lactam (100%; blaEC, blaTEM-1, blaSHV, blaOXA, blaCTX-M-), tetracycline (90.1%; tet(A), tet(B)) and folate synthesis (sul2) antimicrobial resistance genes (ARGs) were most prevalent. The ARGs tet(C), tet(M), tet(32), blaCTX-M-1, blaCTX-M-14, blaOXA-1, dfrA18, dfrA19, catB3, and catB4 were exclusive to human sources, while blaTEM-150, blaSHV-11–12, dfrA12, cmlA1, and cmlA5 were exclusive to beef cattle sources. Frequently encountered virulence factors across all sources included adhesion and type II and III secretion systems, while IncFIB(AP001918) and IncFII plasmids were also common. Specificity and prevalence of ARGs between cattle-sourced and human-sourced presumptive ESBL-EC likely reflect differences in antimicrobial use in cattle and humans. Comparative genomics revealed phylogenetically distinct clusters for isolates from human vs. cattle sources, implying that human infections caused by ESBL-EC in this region might not originate from beef production sources.

The objective of the study was to characterize virulence genes and subtype Escherichia coli O157:H7 and O157:H( 2 ) isolates obtained from a vertically integrated feedlot slaughter plant in Mexico. A total of 1,695 samples were collected from feedlots, holding pens, colon contents, hides, and carcasses. E. coli O157:H7 detection and confirmation was carried out using conventional microbiology techniques, immunomagnetic separation, latex agglutination, and the BAX system. A total of 97 E. coli O157 strains were recovered and screened for key virulence and metabolic genes using multiplex and conventional PCR. Eighty-eight (91.72%) of the strains carried stx2, eae, and ehxA genes. Ten isolates (8.25%) were atypical sorbitol-fermenting strains, and nine were negative for the flicH7 gene and lacked eae, stx1, stx2, and ehxA. One sorbitol-positive strain carried stx2, eae, tir, toxB, and iha genes but was negative for stx1 and ehxA. Pulsed-field gel electrophoresis (PFGE) analysis yielded 49 different PFGE subtypes, showing a high genetic diversity; however, the majority of the typical isolates were closely related (80 to 90% cutoff). Atypical O157 isolates were not closely related within them or to typical E. coli O157:H7 isolates. Identical PFGE subtypes were found in samples obtained from colon contents, feedlots, holding pens, and carcasses. Isolation of a sorbitolfermenting E. coli O157 positive for a number of virulence genes is a novel finding in Mexico. These data showed that genetically similar strains of E. coli O157:H7 can be found at various stages of beef production and highlights the importance of preventing cross-contamination at the pre- and postharvest stages of processing.

This study aimed to better understand the potential public health risk associated with zoonotic pathogens in agricultural fairs and petting zoos in Canada. Prevalence of Salmonella, Shiga toxin-producing Escherichia coli (STEC) O157:H7, and top six non-O157 STEC serogroups in feces (n = 88), hide/feather (n = 36), and hand rail samples (n = 46) was assessed, as well as distributions of antimicrobial resistant (AMR) broad and extended-spectrum β-lactamase (ESBL)-producing E. coli. Prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in pig nasal swabs (n = 4), and Campylobacter, Cryptosporidium, and Giardia in feces was also assessed. Neither Salmonella nor MRSA were detected. Campylobacter spp. were isolated from 32% of fecal samples. Cryptosporidium and Giardia were detected in 2% and 15% of fecal samples, respectively. Only one fecal sample was positive for STEC O157, whereas 22% were positive for non-O157 STEC. Multi-drug resistance (MDR) to antibiotics classified as critically and highly important in human medicine was proportionally greatest in E. coli from cattle feces. The β-lactamase-producing E. coli from pig, horse/donkey feces, and hand rail samples, as well as the STEC E. coli from handrail swabs were MDR. The diversity and prevalence of zoonotic pathogens and AMR bacteria detected within agricultural fairs and petting zoos emphasize the importance of hygienic practices and sanitization with respect to reducing associated zoonotic risks.

The goal of this research was to evaluate the efficacy of a novel rechargeable nonleaching polycationic N-halamine coating applied to stainless steel food contact surfaces to reduce Listeria monocytogenes contamination on ready-to-eat (RTE) foods. Four L. monocytogenes strains were inoculated onto the charged (C; chlorine activated) or noncharged (NC) N-halamine-coated steel coupon surfaces that were either intact or scratched. After inoculation, test surfaces were incubated at 2, 10, and 25°C for 0, 48, and 72 h. L. monocytogenes transfer from coated adulterated surfaces to RTE meat (beef sausages and roast beef) was also tested at 2°C. L. monocytogenes on both intact-C and scratched-C surfaces was significantly reduced at all temperatures; however, in the presence of organic material, these coatings were more effective for reducing L. monocytogenes at 2 and 10°C than at 25°C (P < 0.05). In contrast, on NC intact and scratched surfaces, reduction at 25°C increased (P < 0.05), decreasing the difference in L. monocytogenes levels between charged and noncharged intact and scratched surfaces at this temperature. Overall, greater L. monocytogenes reduction was achieved on intact-C and scratched-C (4.1 ± 0.19 log CFU/cm2) than on intact-NC and scratched-NC (2.3 ± 0.19 log CFU/cm2) surfaces at all temperatures (P < 0.05). The combination of surface condition and chlorine with coupons exposed for 2 h at 2°C in the presence of an organic load (50% meat purge) did not significantly affect the bactericidal efficacy of the N-halamine coating. Regarding transfer to RTE meat, an overall 3.7-log reduction in L. monocytogenes was observed in sausages and roast beef. These findings suggest that a novel rechargeable N-halamine coating on stainless steel surfaces can inactivate L. monocytogenes.

Nontyphoid Salmonella strains are some of the leading causes of foodborne illnesses worldwide; however, there is very limited information on the presence and characteristics of Salmonella in the food production chain in developing countries. In this study, pulsed-field gel electrophoresis (PFGE) was used for molecular subtyping and for monitoring the ecology and transmission of Salmonella isolates in a slaughter facility in Mexico in an attempt to determine specific steps that need to be improved to reduce Salmonella contamination in beef carcasses. A total of 94 isolates from a Salmonella stock culture collection originally obtained from a single vertically integrated feedlot and beef abattoir in Mexico were analyzed. A total of 26 unique PFGE patterns were identified, 38.5% of them corresponding to a single serotype. High concordance (88.4%) was found between serotype and PFGE banding subtype. Salmonella Kentucky and Salmonella Give were the most clonal subtypes in this study, and Salmonella Muenster was the most diverse, with 11 banding patterns identified. A total of 73.7, 70.6, and 85.7% of the PFGE subtypes identified from preevisceration, precooler, and chilled carcasses, respectively, were identified only at those specific points and not at any previous or subsequent steps of the slaughter process, suggesting that each step is in itself a source of Salmonella contamination. Salmonella Mbandaka was more likely to be recovered from feces than from any of the steps of the slaughter process. The genetic diversity and distribution of PFGE subtypes in the processing facility highlight the need to implement antimicrobial interventions and improve sanitation procedures at various points to avoid further Salmonella dissemination into the meat supply.