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[...]to an earlier belief that cattle resisted detrimental effects of Shiga toxins, a comprehensive review of the literature demonstrates that EHEC plays a pivotal role in cattle colonization, with implications for future improved EHEC controls to prevent human disease [1]. A novel mechanism, where the Shiga toxin 1B subunit is sequestered in extracellular vesicles derived from blood cells, demonstrates how the toxin may evade host immune responses [8], providing a better understanding of the mechanism for EHEC pathogenesis. [...]the need for effective surveillance of EHEC to be tailored to local needs was demonstrated by an eight-year survey in South Africa [10].

The use of antimicrobials as growth promotors in livestock has been curtailed by legislative changes in some jurisdictions and the use of chlortetracycline as a growth promoter increased tetracycline resistance in Campylobacter jejeuni, although Canadian beef products were not contaminated by resistant bacteria and food safety risks were minimal [5]. In Brazil, Listeria monocytogenes were present in 12% of beef products sampled and, while antimicrobial resistance was rare, strains were resistant to a common sanitizer used to clean processing equipment, posing a risk to meat hygiene and human health [6]. Inglis, G.D.; Gusse, J.F.; House, K.E.; Shelton, T.G.; Taboada, E.N. Tetracycline Resistant Campylobacter jejuni Subtypes Emanating from Beef Cattle Administered Non-Therapeutic Chlortetracycline are Longitudinally Transmitted within the Production Continuum but are Not Detected in Ground Beef.

Regular usage of NaOH/NaClO disinfectants results in high sodium salt and alkalinity of poultry manure. This study compared three amendments: vegetable waste (V), food waste (F) and mature compost (C) for their ability to improve the composting of NaOH/NaClO-contaminated poultry manure. C compost resulted in the highest compost temperatures (p<0.001) and greatest reduction in OM, TC, TN and NH4-N (p<0.05). C and V composts were more efficient at lowering extractable-Na (ext-Na) and electrical conductivity (EC) than F (p<0.05). Maturity was primarily indicated by NH4-N, EC and ext-Na. Bacterial dynamics was profoundly influenced by NH4-N, EC and TC, with the decrease leading to discriminate genera shift from Sinibacillus and Thiopseudomonas to Brevbacterium, Brachybacterium, and Microbacterium. These findings suggest that mature compost was more desirable amendment than vegetable and food waste in the composting of NaOH/NaClO-contaminated poultry manure, and the decrease of ext-Na indicated compost maturity but did not influence bacterial dynamics.

Pooled feces collected over two years from 1749 transport trailers hauling western-Canadian slaughter cattle were analysed by PCR for detection of Escherichia coli serogroups O26, O45, O103, O111, O121, O145, and O157. Sequential immunomagnetic separation was then used to collect bacterial isolates (n = 1035) from feces positive for target serogroups. Isolated bacteria were tested by PCR to confirm serogroup and the presence of eae, ehxA, stx1, and stx2 virulence genes. Based on PCR screening, serogroup prevalence in feces ranged from 7.0% (O145) to 94.4% (O103) with at least 3 serogroups present in 79.5% of samples. Origin of cattle affected serogroup PCR prevalence and O157 was most prevalent in feces from south-west Alberta (P < 0.001). All serogroups demonstrated seasonal variations in PCR prevalence, with O26, O45, O103, O121, and O157 least prevalent (P < 0.001) in cooler winter months, while uncommon serogroups O111 and O145 increased in prevalence during winter (P < 0.001). However, isolates collected during winter were predominantly from serogroups O103 and O45. No seasonal variation was noted in proportion of isolates which were Shiga toxin containing E. coli (STEC; P = 0.18) or positive for Shiga toxin and eae (enterohemorrhagic E. coli; EHEC; P = 0.29). Isolates of serogroups O111, O145, and O157 were more frequently EHEC than were others, although 37.6-54.3% of isolates from other serogroups were also EHEC. Shiga-toxin genes present also varied by geographic origin of cattle (P < 0.05) in all serogroups except O157. As cattle within feedlots are sourced from multiple regions, locational differences in serogroup prevalence and virulence genes imply existence of selection pressures for E. coli and their virulence in western-Canadian cattle. Factors which reduce carriage or expression of virulence genes, particularly in non-O157 serogroups, should be investigated.

Shiga toxin-producing Escherichia coli (STEC) have been linked to food-borne disease outbreaks. As PCR is routinely used to screen foods for STEC, it is important that factors leading to inconsistent detection of STEC by PCR are understood. This study used whole genome sequencing (WGS) to investigate causes of inconsistent PCR detection of stx 1 , stx 2 , and serogroup-specific genes. Fifty strains isolated from Alberta feedlot cattle from three different studies were selected with inconsistent or consistent detection of stx and serogroup by PCR. All isolates were initially classified as STEC by PCR. Sequencing was performed using Illumina MiSeq® with sample library by Nextera XT. Virtual PCRs were performed using Geneious and bacteriophage content was determined using PHASTER. Sequencing coverage ranged from 47 to 102x, averaging 74x, with sequences deposited in the NCBI database. Eleven strains were confirmed by WGS as STEC having complete stxA and stxB subunits. However, truncated stx fragments occurred in twenty-two other isolates, some having multiple stx fragments in the genome. Isolates with complete stx by WGS had consistent stx 1 and stx 2 detection by PCR, although one also having a stx 2 fragment had inconsistent stx 2 PCR. For all STEC and 18/39 non-STEC, serogroups determined by PCR agreed with those determined by WGS. An additional three WGS serotypes were inconclusive and two isolates were Citrobacter spp. Results demonstrate that stx fragments associated with stx -carrying bacteriophages in the E . coli genome may contribute to inconsistent detection of stx 1 and stx 2 by PCR. Fourteen isolates had integrated stx bacteriophage but lacked complete or fragmentary stx possibly due to partial bacteriophage excision after sub-cultivation or other unclear mechanisms. The majority of STEC isolates (7/11) did not have identifiable bacteriophage DNA in the contig(s) where stx was located, likely increasing the stability of stx in the bacterial genome and its detection by PCR.

Feed contaminated with the mycotoxin deoxynivalenol (DON) can negatively impact livestock health and performance. Bacteria capable of degrading DON present a method of mitigating its harmful effects. This study aimed to identify microbial consortia from soil samples that could degrade DON. Soil from central (Lacombe, LA) and southern (Lethbridge, LE) Alberta were used as microbial inoculant. The soils were mixed with DON-contaminated wheat (18 ppm/kg) on day 0, and each soil type was divided into triplicate pots (180 g) and placed in a controlled environment for 32 d. Control pots of each soil type were included, which contained no DON-contaminated wheat. On days 0, 7, 14, and 32, 1 g subsamples were collected from pots, serially diluted in a limited medium containing DON (10 µg/mL) as the only carbon source, and incubated for 2 weeks (30 °C). DNA was extracted from the pots across time, as well as the subsample consortia grown in DON-amended medium, and was analyzed for bacterial changes after 16S rRNA gene sequencing. The relative abundance of bacterial genera in soil samples after enrichment with DON-contaminated wheat increased across time compared to the baseline day 0 time point. DON-degrading activity (26%) was only detected in LA soil suspension on day 7, and was highest after 14 days of incubation. The most abundant bacteria in the LA DON-degrading consortia belonged to the Pseudomonas (8.8%), Delftia (7.4%), Acinetobacter (6.4%), Comamonas (5.7%), Stenotrophomonas (5.5%), Shinella (5.5%), Ensifer (5.1%), Agrobacterium (5.0%), Achromobacter (4.7%), and Rhizobium (3.7%) genera. Pseudomonas aeruginosa (n = 9) and Serratia liquefaciens (n = 3) strains isolated from the LA consortia did not degrade DON. Overall, this study shows that the soil contained bacteria capable of degrading DON; however, variation existed depending on the soil’s source.

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.

Several areas of the world suffer a notably high incidence of Shiga toxin-producing Escherichia coli . To assess the impact of persistent cross-species transmission systems on the epidemiology of E. coli O157:H7 in Alberta, Canada, we sequenced and assembled E. coli O157:H7 isolates originating from collocated cattle and human populations, 2007–2015. We constructed a timed phylogeny using BEAST2 using a structured coalescent model. We then extended the tree with human isolates through 2019 to assess the long-term disease impact of locally persistent lineages. During 2007–2015, we estimated that 88.5% of human lineages arose from cattle lineages. We identified 11 persistent lineages local to Alberta, which were associated with 38.0% (95% CI 29.3%, 47.3%) of human isolates. During the later period, six locally persistent lineages continued to be associated with human illness, including 74.7% (95% CI 68.3%, 80.3%) of reported cases in 2018 and 2019. Our study identified multiple locally evolving lineages transmitted between cattle and humans persistently associated with E. coli O157:H7 illnesses for up to 13 y. Locally persistent lineages may be a principal cause of the high incidence of E. coli O157:H7 in locations such as Alberta and provide opportunities for focused control efforts.

Over a two-year period, Mannheimia haemolytica (MH; n = 113), Pasteurella multocida (PM; n = 47), Histophilus somni (HS; n = 41) and Mycoplasma bovis (MB; n = 227) were isolated from bovine lung tissue at necropsy from cattle raised conventionally (CON, n = 29 feedlots) or without antimicrobials [natural (NAT), n = 2 feedlots]. Excluding MB, isolates were assayed by PCR to detect the presence of 13 antimicrobial resistance (AMR) genes and five core genes associated with integrative and conjugative elements (ICEs). Antimicrobial susceptibility phenotypes and minimum inhibitory concentrations (MICs, µg/mL) were determined for a subset of isolates (MH, n = 104; PM, n = 45; HS, n = 23; and MB, n = 61) using Sensititre analyses. A subset of isolates (n = 21) was also evaluated by whole-genome sequencing (WGS) based on variation in AMR phenotype. All five ICE core genes were detected in PM and HS by PCR, but only 3/5 were present in MH. Presence of mco and tnpA ICE core genes in MH was associated with higher MICs (p < 0.05) for all tetracyclines, and 2/3 of all macrolides, aminoglycosides and fluoroquinolones evaluated. In contrast, association of ICE core genes with MICs was largely restricted to macrolides for PM and to individual tetracyclines and macrolides for HS. For MH, the average number of AMR genes markedly increased (p < 0.05) in year 2 of the study due to the emergence of a strain that was PCR positive for all 13 PCR-tested AMR genes as well as two additional AMR genes (aadA31 and blaROB-1) detected by WGS. Conventional management of cattle increased (p < 0.05) MICs of tilmicosin and tulathromycin for MH; neomycin and spectinomycin for PM; and gamithromycin and tulathromycin for MB. The average number of PCR-detected AMR genes in PM was also increased (p < 0.05) in CON mortalities. This study demonstrates increased AMR especially to macrolides by bovine respiratory disease organisms in CON as compared to NAT feedlots and a rapid increase in AMR following dissemination of strain(s) carrying ICE-associated multidrug resistance.

Often Escherichia coli are harmless and/or beneficial bacteria inhabiting the gastrointestinal tract of livestock and humans. However, Shiga toxin-producing E. coli (STEC) have been linked to human disease. Cattle are the primary reservoir for STEC and STEC \"super-shedders\" are considered to be a major contributor in animal to animal transmission. Among STEC, O157:H7 is the most recognized serotype, but in recent years, non-O157 STEC have been increasingly linked to human disease. In Argentina and Germany, O178 is considered an emerging pathogen. Our objective was to compare populations of E. coli O178, O157, shiga toxin 1 and 2 in western Canadian cattle feces from a sampling pool of ~80,000 beef cattle collected at two slaughterhouses. Conventional PCR was utilized to screen 1,773 samples for presence/absence of E. coli O178. A subset of samples (n = 168) was enumerated using droplet digital PCR (ddPCR) and proportions of O178, O157 and shiga toxins 1 & 2 specific-fragments were calculated as a proportion of generic E. coli (GEC) specific-fragments. Distribution of stx1 and stx2 was determined by comparing stx1, stx2 and O157 enumerations. Conventional PCR detected the presence of O178 in 873 of 1,773 samples and ddPCR found the average proportion of O178, O157, stx1 and stx2 in the samples 2.8%, 0.6%, 1.4% and 0.5%, respectively. Quantification of stx1 and stx2 revealed more virulence genes than could be exclusively attributed to O157. Our results confirmed the presence of E. coli O178 in western Canadian cattle and ddPCR revealed O178 as a greater proportion of GEC than was O157. Our results suggests: I) O178 may be an emerging subgroup in Canada and II) monitoring virulence genes may be a more relevant target for food-safety STEC surveillance compared to current serogroup screening.