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Porcine kobuviruses are widely distributed in swine, but the clinical significance of these viruses remains unclear, since they have been associated with both diarrheic and healthy pigs. In addition, there is a paucity of data on Kobuvirus prevalence in Canadian pig herds. In this study, a total of 181 diarrheic and healthy piglets were monitored and sampled on four occasions, intended to represent the different stages of production. The piglets were sampled at the nursing farms (birth to weaning stage), at the nursery farms (post-weaning stage), and at finishing farms (at the beginning and the end of the fattening stage). Fecal and environmental samples were collected during each life stage. Following viral extraction, Kobuvirus detection by RT-PCR was conducted, and positive samples were sequenced. During the late-nursing stage (6–21 days old), piglets with diarrhea shed more Kobuvirus than healthy individuals. Piglets shed more Kobuvirus during the post-weaning stage (nursery farms) than during any of the other life stages. This was evidenced in individual samples as well as in environmental samples. Over 97% of the sampled piglets shed Kobuvirus at least once in their lifetime. All piglets shedding a Kobuvirus strain or mix of strains at the nursing stage did not appear to shed another porcine kobuvirus strain at a later life stage. Overall, our findings throw light on Kobuvirus shedding dynamics and their potential role in neonatal diarrhea at the nursing stage, which appears to be the point of entry for kobuviruses into swine production systems.

The Canadian Food Inspection Agency (CFIA), as part of the modernization of its inspection system, is developing a risk assessment model with the goal of evaluating the most important food safety-related risk factors associated with the various types of food establishments under its jurisdiction. Ultimately, this new risk assessment model will assist CFIA in allocating its inspection resources according to the level of risk associated with each establishment. As a first step, key risk factors need to be identified. The objective of the current study was thus to identify and evaluate the importance of food safety-related risk factors that could be included in a risk assessment model. An initial literature search was used to identify these risk factors. Thereafter, 75 Canadian food safety experts were asked to assess the relative importance of the 155 identified risk factors with respect to food safety. This step was carried out using a web-based questionnaire in which risk factors had to be assessed by experts using a ten-point Likert-type scale. One risk factor (“management commitment”) received a median score of 10, while median score of 9, 8.5 and 8, and below 8 were attributed to 42, 77 and 35 of the risk factors listed, respectively. A company's commitment towards food safety was identified as an important risk factor contributing to the magnitude of the risk attributed to a food establishment. Many factors related to the performance of an establishment's preventive control plans were also scored high by experts, such as sanitation, hygiene practices, calibration and employee training. The median interquartile range in risk factor scoring generally did not exceed 2, and the respondent profile (work experience, employer and/or commodity-based expertise) did not have a strong influence on score results based on Kruskal–Wallis tests. This observation suggests that, despite their different backgrounds, experts’ opinions were relatively consistent and comparable when scoring the impact of various risk factors towards food safety. These survey outcomes represent useful information to aid in the design and implementation of risk assessment models for food establishments.

International audience; AIM: To observe the transfer of the digestive microbiota from sow to piglet, describe the impact of the sow's Salmonella shedding on this transfer and identify transferred populations that could be associated with the future Salmonella status of the piglets.METHODS AND RESULTS: Salmonella shedding status of 19 sows was determined at the beginning and end of gestation. Four piglets were randomly selected from each sow. Using MiSeq, the microbiotas of the sows at the end of gestation and of their piglets 1 day before weaning were described. Results showed that the Salmonella shedding of the sows, the birth mother, the lairage room, the parity and the contamination of the lairage environment were associated to the microbiota of the piglets (permanova P < 0·05). Several genera were associated with piglets born from negative or positive sows.CONCLUSION: There is a link between the microbiota of the sows at the end of gestation and the microbiota of their piglets at weaning, and the Salmonella shedding of the sow is associated with the microbiota of the piglets.SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella status of the sows affects the microbiota of their piglets and could affect the long-term Salmonella colonization resistance of these animals and their health.

Lactation, feeding conditions, microbial interventions and piglet growth in the first few weeks of life have important impact on the intestinal microbiota establishment and immune system development of piglets. Indeed, colostrum and milk contain various bioactive components such as immune factors, antimicrobial peptides and oligosaccharides that contribute to maintain intestinal homeostasis and regulate interactions between microbiota and host immune system. Recent results revealed that low birth weight piglet (LBWP) with poor weight gain during the first two weeks of life develop different intestinal microbiota and immune response profiles compared to high BWP (HBWP) littermates. Consequently, piglets within litters may have different resilience to infections after weaning and benefit from feed additives in a specific manner. A study has been performed to evaluate the potential of bovine colostrum extract (BC) as replacement to plasma proteins for improving gut health and resilience to Salmonella infection in piglets. Results revealed that in weaned piglets fed BC, intestinal microbiota was differently modulated and bacterial dysbiosis induced by Salmonella was restored faster. Moreover, expression of genes involved in innate immunity such as β-defensin-2 and glutathione peroxidase-2 was respectively down- and up-regulated in BC fed piglets. A combination of dietary supplementation with BC, cupper and vitamins A and D has also been tested in LBWP and HBWP, and there is clear evidence that BC in combination with other feed additives promote growth and gut health in both LBWP and HBWP. The porcine intestinal epithelial cell line IPEC-J2 was used to better understand the functional properties of BC. Results indicated that BC improves wound healing, enhances barrier function and modulates the expression of several genes involved in innate immune response. Finally, as microbial intervention, the potential of fecal transplantation to modulate intestinal microbiota and immune system development of piglets is under investigation and will be discussed.

Clostridium perfringens ranks among the three most frequent bacterial pathogens causing human foodborne diseases in Canada, and poultry meat products are identified as a source of infection for humans. The objective of the current study was to estimate the proportion of broiler chicken flocks, carcasses and various environmental samples from critical locations of the slaughter plant positive for the presence of C. perfringens enterotoxin encoding gene (cpe). From the 16 visits conducted, 25% of the 79 flocks sampled, 10% of the 379 carcasses sampled and 5% of the 217 environmental samples collected were found positive for cpe. The proportion of cpe-positive carcasses was statistically different between surveyed plants, with 17.0% for one abattoir and 2.2% for the other. For the most contaminated plant, cpe-positive carcasses were identified at each step of the processing line, with prevalence varying between 10.0% and 25.0%, whereas this prevalence varied between 0% and 25.0% for the environmental surfaces sampled. Based on the results obtained, enterotoxigenic C. perfringens strains could potentially represent a risk for the consumer.

Post-weaning diarrhea (PWD) is one of the most serious threats for the swine industry worldwide. It is commonly associated with the proliferation of enterotoxigenic Escherichia coli in the pig intestine. Colistin, a cationic antibiotic, is widely used in swine for the oral treatment of intestinal infections caused by E. coli, and particularly of PWD. However, despite the effectiveness of this antibiotic in the treatment of PWD, several studies have reported high rates of colistin resistant E. coli in swine. Furthermore, this antibiotic is considered of very high importance in humans, being used for the treatment of infections due to multidrug-resistant (MDR) Gram-negative bacteria (GNB). Moreover, the recent discovery of the mcr-1 gene encoding for colistin resistance in Enterobacteriaceae on a conjugative stable plasmid has raised great concern about the possible loss of colistin effectiveness for the treatment of MDR-GNB in humans. Consequently, it has been proposed that the use of colistin in animal production should be considered as a last resort treatment only. Thus, to overcome the economic losses, which would result from the restriction of use of colistin, especially for prophylactic purposes in PWD control, we believe that an understanding of the factors contributing to the development of this disease and the putting in place of practical alternative strategies for the control of PWD in swine is crucial. Such alternatives should improve animal gut health and reduce economic losses in pigs without promoting bacterial resistance. The present review begins with an overview of risk factors of PWD and an update of colistin use in PWD control worldwide in terms of quantities and microbiological outcomes. Subsequently, alternative strategies to the use of colistin for the control of this disease are described and discussed. Finally, a practical approach for the control of PWD in its various phases is proposed.

Objective To evaluate whether pig farms interconnected within the same cooperative share similar Salmonella contamination patterns. Setting Ten finishing pig farms within a 100 km radius of a common slaughterhouse were selected. Their inclusion was based on their association to the same cooperative and the sharing of common resources: piglets, feed, swine transporters, slaughterhouse, technicians and veterinarians. Procedure Each farm was visited three times over a 10-month period. Pig faeces, the barn front door handle, the feed pipeline, mobile objects (shovel, balance and pig board), the landing stage, the concrete slab of the feed bins, the tire tracks left on the pathways by the animal feed truck, the pig delivery truck and the carcase knacker truck and the mudguards and cabin carpets of the veterinarian and technician vehicles on their arrival at the farm were all analysed for the presence of Salmonella. Results All farms were not equally contaminated with Salmonella. Whereas some farms yielded up to 12 Salmonella isolates, other farms were Salmonella free. Some locations, most notably the landing stage, were more contaminated than others. Salmonella contamination was dynamic in time. Some contaminations seen on farms, on specific locations on the first visit, had disappeared on the second and third visits, but new contaminations were detected on different locations. Conclusions Contamination with Salmonella was not disseminated through the network of the 10 pig farms interconnected within the same cooperative but was rather most often restricted in time to specific locations on specific farms.

Campylobacter jejuni is a zoonotic agent responsible for the foodborne gastroenteritis campylobacteriosis. Control of C. jejuni load in the poultry primary production is recognized as an avenue to reduce human exposure to the pathogen. As for now, no commercially applicable control methods exist at the farm. Several studies tested egg yolk powders, potentiated or not against C. jejuni, as feed additives for chicken and suggested that the quantity and quality of the antibodies presence in the yolk are determinant factors for the full success of this approach. Unfortunately, data from these studies inconsistently showed a reduction of cecal C. jejuni carriage. Our first goal wwas to characterize (quantification by ELISA, agglutination test, bacterial antigen recognition profiles by Western blot, bactericidal effect by serum killing assays and C. jejuni mobility by soft agar migation) the antibodies extracted from egg yolk powders originating from different egg production protocols. Secondly, these powders were microencapsulated and recharacterized. Finally the protected powders were tested as a feed additive to destabilize C. jejuni colonization in an in vivo assay. Despite the in vitro results indicating the ability of the egg yolk powders to recognize Campylobacter and potentially alter its colonization of the chicken caecum, these results were not confirmed in the in vivo trial despite that specific caecal IgY directed toward Campylobacter were detected in the groups receiving the protected powders. More research is needed on Campylobacter in order to effectively control this pathogen at the farm.

This report describes various Salmonella serovars which were found on often overlooked locations in a pig farm/slaughterhouse interface. These include slaughterhouse yard pathways and mudguards and carpets of transport trucks arriving at and departing from production sites.

The aim of this study was to investigate the evolution with time of ceftiofur-resistant Escherichia coli clinical isolates from pigs in Québec, Canada, between 1997 and 2012 with respect to pathotypes, clones and antimicrobial resistance. Eighty-five ceftiofur-resistant E. coli isolates were obtained from the OIE (World Organisation for Animal Health) Reference Laboratory for Escherichia coli. The most prevalent pathovirotypes were enterotoxigenic E. coli (ETEC):F4 (40%), extraintestinal pathogenic E. coli (ExPEC) (16.5%) and Shiga toxin-producing E. coli (STEC):F18 (8.2%). Susceptibility testing to 15 antimicrobial agents revealed a high prevalence of resistance to 13 antimicrobials, with all isolates being multidrug-resistant. blaCMY-2 (96.5%) was the most frequently detected β-lactamase gene, followed by blaTEM (49.4%) and blaCTX-M (3.5%). Pulsed-field gel electrophoresis (PFGE) applied to 45 representative E. coli isolates revealed that resistance to ceftiofur is spread both horizontally and clonally. In addition, the emergence of extended-spectrum β-lactamase-producing E. coli isolates carrying blaCTX-M was observed in 2011 and 2012 in distinct clones. The most predominant plasmid incompatibility (Inc) groups were IncFIB, IncI1, IncA/C and IncFIC. Resistance to gentamicin, kanamycin and chloramphenicol as well as the frequency of blaTEM and IncA/C significantly decreased over the study period, whereas the frequency of IncI1 and multidrug resistance to seven antimicrobial categories significantly increased. These findings reveal that extended-spectrum cephalosporin-resistant porcine E. coli isolates in Québec belong to several different clones with diverse antimicrobial resistance patterns and plasmids. Furthermore, blaCMY-2 was the major β-lactamase gene in these isolates. From 2011, we report the emergence of blaCTX-M in distinct clones.

Viral contamination along the production chain is a significant concern in both food safety and livestock health. Pigs have been reported to act as a reservoir for zoonotic viruses, sometimes emerging ones, and epidemiological studies have shown direct links between the consumption of uncooked pork offal and cases of hepatitis caused by the hepatitis E virus (HEV) genotype 3 in humans. The presence of HEV in swine herds has been reported, but its dissemination in pork production environments is still unknown. To investigate viral contamination sources in the swine industry, 452 environment and fecal samples, including samples from livestock transportation vehicles, were collected over a period of 11 months from ten farms and one slaughterhouse that together represent a single production network. Hepatitis E virus RNA was detected by nested RT-PCR in 32 samples from both inside and outside farm buildings, on trucks, and, mostly, from fomites collected in the slaughterhouse yard, such as on a utility vehicle. Phylogenetic analysis showed a wide diversity of HEV genotype 3 strains, similar to human and swine strains previously found. According to the results of this study, the movements of trucks and utility vehicles might play an important role in HEV dissemination on a slaughterhouse site and throughout an entire network.

Aims To investigate the mechanisms leading to an increase in the prevalence of blaCMY -2 conferring resistance to ceftiofur in pigs receiving a feed medicated with chlortetracycline and penicillin, and to examine the effect of supplementation with a clay mineral on this phenomenon. Methods and results In 138 blaCMY -2 -positive Escherichia coli isolates from faeces of pigs receiving feed supplemented or not with 2% clinoptilolite, from day 2 to day 28 after weaning, isolates from the two groups differed significantly with respect to their phylogenetic group: phylotype A predominated in the supplemented group, whereas phylotypes B1 and D predominated in the control group, as determined by PCR. In 36 representative isolates, pulsed-field gel electrophoresis and antimicrobial susceptibility testing revealed that the blaCMY -2 -positive E. coli isolates were polyclonal with diverse antimicrobial resistance patterns and blaCMY -2 -carrying plasmids of incompatibility (Inc) groups, A/C, I1 and ColE were observed in transformants as detected by PCR. Enterobacter cloacae possessing blaCMY -2 -carrying IncA/C plasmids were found in the pens before introduction of this batch of pigs. The blaCMY -2 -positive E. coli isolates were more clonally diverse in the control group than the supplemented group. Conclusions The blaCMY -2 gene appears to have spread both horizontally and clonally in this batch of pigs and may have spread from previous batches of pigs via plasmids carried by Ent. cloacae and expanded in animals of the present batch in the presence of the selection pressure due to administration of chlortetracycline and penicillin in the feed. Feed supplementation may have an effect on clonal diversity of blaCMY -2 -positive isolates. Significance and impact of the study Implementation of improved hygiene measures, decreased administration of certain antimicrobials on farm and feed supplementation with certain ingredients may limit antimicrobial resistance spread between and within batches of animals.

Feed characteristics may influence the bacterial community composition and metabolic activities in the pig gastrointestinal tract, known to be associated with positive effects on the gut. Use of mash feed is associated with reduced excretion, but little is known of its effect on the population or of the mechanism of action. Our objectives were to assess the effect of feed texture combined with feed particle size on VFA profiles and levels, total count, and the presence of genes encoding virulence factors of pathogenic strains in the digestive tract along with their impact on pig performance of fattening pigs. Pigs ( = 840) on a commercial farm received mash or pellet diets of different particle sizes during the fattening period. Caecal and colon contents from 164 pigs were sampled at the slaughterhouse for enumeration of by quantitative PCR (qPCR) and for VFA quantification by capillary gas chromatography. The gene was used to enumerate total . Improved pig performances associated with pellet texture and a 500-μm size were observed. Caecal ( = 0.02) and colon ( < 0.01) propionic acid concentrations were lower for pigs receiving pellet rather than mash feed. Similarly, caecal ( = 0.01) and colon ( < 0.001) butyric acid concentrations were also lower for pigs receiving pellet rather than mash feed, as determined by capillary gas chromatography. Moreover, caecal ( = 0.03) and colon ( < 0.001) butyric acid concentrations were higher for pigs receiving a feed with a 1,250-μm particle size rather than a 500-μm particle size. On the other hand, total caecal and colon levels were higher for pigs receiving pellet feed than for those receiving mash feed. For total enumeration, caecal ( < 0.01) and colon ( < 0.01) gene copies were higher for pigs receiving pellet rather than mash feed. No effect of particle size on fatty acid concentrations or on numbers was observed. Virulence gene quantification revealed no trend. Taken together, results showed that mash feed is associated with lower growth performance but with favorable intestinal changes linked to VFA levels and reduction in the intestine.

This study was carried out to evaluate the prevalence of Salmonella serogroups and serotypes and their virulence gene carriage in pig fecal samples from farms and slaughterhouses in some southern provinces of Vietnam. The presence of Salmonella was assessed based on culture enrichment of the collected samples and biochemical and serological analyses; 27.7% (51) of 184 samples were posititve for Salmonella. Based on the availability of antisera, serogroups were determined for 61% (31) of 51 isolates. Twenty isolates belonging to Salmonella serotypes Typhimurium (10 isolates), Anatum (8 isolates), Senftenberg (7 isolates), Paratyphi B (3 isolates), Paratyphi A (1 isolate), Montevideo (1 isolate), and Saintpaul (1 isolate) were further characterized by a multiplex PCR protocol targeting Salmonella invasion A and virulence plasmid C genes ( invA and spvC, respectively). Individual PCR assays were developed to detect genes for Salmonella enterotoxin ( stn) , Salmonella outer protein B ( sopB), and Salmonella fimbriae ( pef). Various carriage patterns were identified among tested isolates. The invA and sopB genes were found in all isolates, and the stn gene was found in 95% of the isolates. The spvC gene was found in only 5% of the Salmonella Typhimurium isolates. None of the isolates were positive for the pef gene. Among all isolates, the predominant genotypic virulence profile (virulotype) was characterized by the concomitant presence of invA, sopB, and stn in carrier strains. In contrast, two virulotypes comprising either invA, sopB, spvC, and stn or invA and sopB were identified for the Salmonella Typhimurium isolates. Virulotypes made up of multiple virulence genes were predominant in most Salmonella strains tested in this study, indicating that pigs might act as a reservoir for these virulent strains.

The aim of the present study was to investigate the in vitro gastric stability of colistin sulfate (CS) and its antimicrobial activity against Escherichia coli and to study the impact of ETEC O149: F4 (K88) infection in pigs on CS intestinal absorption. The stability profile of CS was evaluated in a simulated gastric fluid (SGF). Antimicrobial activity of CS and its degradation products were examined in a 96-well polystyrene microplate model. The effect of experimental infection with ETEC O149: F4 on CS intestinal absorption was determined by quantification of CS systemic concentration using a validated LC–MS/MS method. A rapid degradation of CS accompanied by an increase in CS antimicrobial activity by comparison with non-degraded CS (P < 0.0001) was observed in SGF. Additionally, CS levels were not quantifiable in systemic circulation using a highly sensitive method and concurrent oral challenge did not affect CS absorption in an induction model of subclinical post-weaning diarrhea (PWD).

Following the 2008 Canadian listeriosis outbreak associated with ready-to-eat (RTE) meat products, regulations on the presence of Listeria monocytogenes in RTE food production facilities were modified by Health Canada, confirming the need to control this pathogen, not only in the final product but also in the plant environment. Information on the occurrence of this microorganism during the early steps of production, such as the slaughtering process and in the cutting area, is scarce in Canada. In this study, we sampled different production steps in a slaughtering and cutting plant in the province of Quebec over a 2-year period. The lairage pens, representative areas of the slaughter line, and cutting zones were targeted after their respective cleaning procedures. A total of 874 samples were analyzed for the presence of L. monocytogenes. Characterization was done by first genoserogrouping the isolates using multiplex PCR and then using a pulsed-field gel electrophoresis approach. L. monocytogenes was detected throughout all production stages. The 108 positive samples found were analyzed further, and we established that there were 4 different serogroups, with serogroup IIb being the most prevalent. The results of pulsed-field gel electrophoresis analysis showed a significant decrease in the diversity of strains from the first areas of the plant to the cutting room (10 pulsotypes in 13 positive samples in lairage and 9 in 86 positive samples in cutting) and also showed the overrepresentation of a single predominant strain in the cutting room environment (type 1, representing 96.1% of the isolates). Biofilm formation analysis of the strains cannot exclusively explain the transitions we observed. A strong genotypic similarity between strains isolated in the early production areas and some strains in the cutting room was shown. These results support the need for better surveillance of L. monocytogenes prior to RTE food production in order to design control strategies that are better adapted from a public health perspective.

Pork meat is estimated to be responsible for 10–20% of human salmonellosis cases in Europe. Control strategies at the farm could reduce contamination at the slaughterhouse. One of the targeted sectors of production is maternity, where sows could be Salmonella reservoirs. The aim of this study was to assess the dynamics of shedding of Salmonella in terms of variation in both shedding prevalence and strains excreted during gestation in Quebec’s maternity sector. The evolution of the fecal microbiota of these sows during gestation was also assessed to detect bacterial populations associated with these variations. A total of 73 sows both at the beginning and the end of the gestation were randomly selected and their fecal matter was analyzed. Salmonella detection was conducted using a method that includes two selective enrichment media (MSRV and TBG). Nine isolates per positive samples were collected. Among the 73 sows tested, 27 were shedding Salmonella. Sows in the first third of their gestation shed Salmonella significantly more frequently (21/27) than those in the last third (6/46) (χ2 P < 0.05). The shedding status of 19 of the sows that were previously sampled in the first third of their gestation was followed, this time in the last third of their gestation, which confirmed reduction of shedding. Using 16S rRNA gene sequencing and qPCR, significant differences between the fecal flora of sows at the beginning and the end of the gestation, shedding Salmonella or not and with different parity number were detected. Using MaAsLin, multiple OTUs were found to be associated with the time of gestation, the status of Salmonella excretion and parity number. Some of the identified taxa could be linked to the reduction of the shedding of Salmonella at the end of gestation. In this study, we showed that the level of Salmonella shedding was variable during gestation with significantly higher shedding at the beginning rather than at the end of gestation. We also observed for the first time a significant change in the microbiota during sow gestation and identified interesting taxa which could be linked to a reduced Salmonella shedding.

The introduction of Listeria monocytogenes into the food production chain is a concern, with numerous grouped cases of listeriosis associated with milk-derived or pork-derived products have been documented. Management of this zoonotic pathogen considers all strains as an equal risk. Recently, a new perspective for characterisation of strain virulence was introduced with the discovery of the unaltered sequence of InlA as a determinant of strain virulence; this has also been reported as an infrequent finding among so-called environmental strains, that is, strains isolated from food or from surfaces in food industries. The aim of this study was to differentiate L monocytogenes strains isolated from animal cases versus those from human cases and to differentiate clinical strains from environmental ones using a Caenorhabditis elegans virulence testing model. In Quebec in 2013/2014, the surveillance of L monocytogenes clinical isolates registered a total of 20 strains of animal origin and 16 pulsed-field gel electrophoresis types isolated from human cases. The mixed PCR multiplex agglutination protocol used for geno-serotyping clearly discriminated genogroup IVB strains from bovine and human origins. The presence of a premature stop codon single nucleotide polymorphism in the inlA gene sequence in clinical strains and the identical behaviour of particular strains in the C elegans model are discussed in this paper from the perspective of industrial management of L monocytogenes risk.

International audience; Faced with ever-increasing demand, the industrial production of food animals is under pressure to increase its production. In order to keep productivity, quality, and safety standards up while reducing the use of antibiotics, farmers are seeking new feed additives. In chicken production, one of these additives is selenium. This element is expected to confer some advantages in terms of animal health and productivity, but its impact on chicken intestinal microbiota as well as on the carriage of foodborne pathogens is unknown. In this study, chickens raised in a level 2 animal facility were fed or not 0.3 ppm of in-feed selenium-yeast until 35 days of age and were inoculated or not with the foodborne pathogen Campylobacter jejuni at the age of 14 days. At the end of the study, body weight, seric IgY, intestinal IgA, seric gluthatione peroxydase activity, the caecal microbiota (analyzed by MiSeq 16S rRNA gene sequencing), and C. jejuni caecal levels were analyzed. The experiment was completely replicated twice, with two independent batches of chickens. This study revealed that, for healthy chickens raised in very good hygienic conditions, selenium-yeast does not influence the bird's body weight and lowers their seric gluthatione peroxidase activity as well as their intestinal IgA concentrations. Furthermore, selenium-yeast did not modify the caecal microbiota or the colonization of C. jejuni. The results also showed that C. jejuni colonization does not impact any of the measured chicken health parameters and only slightly impacts the caecal microbiota. This study also clearly illustrated the need for true biological replication (independent animal trials) when assessing the microbiota shifts associated with treatments as the chickens microbiotas clearly clustered according to study replicate.

The Canadian Food Inspection Agency (CFIA) is developing a risk assessment model for food establishments. Previous research on the significance of food safety risk factors determined by literature review and expert advice served as the bases for the current study, to further refine, discriminate and select the most important criteria to be included in the model. This process considered the availability of data sources, the clarity and measurability of the selected factors, undertook the elimination of lower-rated risk factors and grouped those with similar focus of attention, enabling the selection of a final list of risk factors for the model. A method of assessment for the remaining factors was then proposed to allow the quantification of individual risk factors within the model. From the 155 risk factors initially identified, 17 consolidated factors were kept and will be considered for the development of the risk assessment model.

Modern swine production systems represent complex and dynamic networks involving numerous stakeholders. For instance, livestock transporters carry live animals between fattening sites, abattoirs, and other premises on a daily basis. This interconnected system may increase the risk of microbial spread within and between networks, although little information is available in that regard. In the present study, a swine network composed of 10 finishing farms, one abattoir, and three types of stakeholders (veterinarians, livestock transporters, and nutritional technicians) in Quebec, Canada, was selected to investigate specific vectors and reservoirs of enteric viruses. Environmental samples were collected from the premises over a 12-month period. Samples were screened using targeted reverse transcription-PCR and sequencing of two selected viral markers, group A rotaviruses (RVA) and porcine astroviruses (PoAstV), both prevalent and genetically heterogeneous swine enteric viruses. The results revealed frequent contamination of farm sites (21.4 to 100%), livestock transporter vehicles (30.6 to 68.8%) and, most importantly, the abattoir yard (46.7 to 94.1%), depending on the sample types. Although high levels of strain diversity for both viruses were found, identical PoAstV and RVA strains were detected in specific samples from farms, the abattoir yard, and the livestock transporter vehicle, suggesting interconnections between these premises and transporters. Overall, the results from this study underscore the potential role of abattoirs and livestock transport as a reservoir and transmission route for enteric viruses within and between animal production networks, respectively. IMPORTANCE Using rotaviruses and astroviruses as markers of enteric contamination in a swine network has revealed the potential role of abattoirs and livestock transporters as a reservoir and vectors of enteric pathogens. The results from this study highlight the importance of tightening biosecurity measures. For instance, implementing sanitary vacancy between animal batches and emphasizing washing, disinfection, and drying procedures on farms and for transportation vehicles, as well as giving limited access and circulation of vehicles throughout the production premises, are some examples of measures that should be applied properly. The results also emphasize the need to closely monitor the dynamics of enteric contamination in the swine industry in order to better understand and potentially prevent the spread of infectious diseases. This is especially relevant when a virulent and economically damaging agent is involved, as seen with the recent introduction of the porcine epidemic diarrhea virus in the country.

Background Enterotoxigenic Escherichia coli (ETEC) strains producing multiple enterotoxins are important causes of post-weaning diarrhea (PWD) in pigs. The aim of the present study was to investigate the fecal presence of ETEC enterotoxin as well as F4 and F18 genes as an indicator of colistin sulfate (CS) efficacy for treatment of PWD in pigs. Forty-eight piglets were weaned at the age of 21 days, and were divided into four groups: challenged treated, challenged untreated, unchallenged treated, and unchallenged untreated. Challenge was performed using 109 CFU of an ETEC: F4 strain, and treatment was conducted using oral CS at the dose of 50,000 IU/kg. The fecal presence of genes encoding for STa, STb, LT, F4 and F18 was detected using PCR. Results The PCR amplification of ETEC virulence genes showed that nearly 100% of pigs excreted genes encoding for STa and STb toxins in the feces before the challenge. These genes, in the absence of the gene encoding F4, were considered as a marker for F4-negative ETEC. One day after ETEC: F4 oral challenge pigs in the two challenged groups excreted the genes encoding LT and F4 in the feces. These genes were considered as a marker for F4-positive ETEC. However, the gene encoding F18 was not detected in any fecal samples of the 4 groups throughout the experiment. After only 3 days of successive oral treatment with CS, a significant reduction in both the F4-positive and negative ETEC populations was observed in the challenged treated group compared to the challenged untreated group (p

Colistin (Polymyxin E) is one of the few cationic antimicrobial peptides commercialized in both human and veterinary medicine. For several years now, colistin has been considered the last line of defense against infections caused by multidrug-resistant (MDR) Gram-negative such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae. Colistin has been extensively used orally since the 1960s in food animals and particularly in swine for the control of Enterobacteriaceae infections. However, with the recent discovery of plasmid-mediated colistin resistance encoded by the mcr-1 gene and the higher prevalence of samples harboring this gene in animal isolates compared to other origins, livestock has been singled out as the principal reservoir for colistin resistance amplification and spread. Co-localization of the mcr-1 gene and Extended-Spectrum- β-lactamase (ESBL) genes on a unique plasmid has been also identified in many isolates from animal origin. The use of colistin in pigs as a growth promoter and for prophylaxis purposes should be banned, and the implantation of sustainable measures in pig farms for microbial infection prevention should be actively encouraged and financed. The scientific research should be encouraged in swine medicine to generate data helping to reduce the exacerbation of colistin resistance in pigs and in manure. The establishment of guidelines ensuring a judicious therapeutic use of colistin in pigs, in countries where this drug is approved, is of crucial importance. The implementation of a microbiological withdrawal period that could reduce the potential contamination of consumers with colistin resistant bacteria of porcine origin should be encouraged. Moreover, the management of colistin resistance at the human-pig-environment interface requires the urgent use of the One Health approach for effective control and prevention. This approach needs the collaborative effort of multiple disciplines and close cooperation between physicians, veterinarians, and other scientific health and environmental professionals. This review is an update on the chemistry of colistin, its applications and antibacterial mechanism of action, and on Enterobacteriaceae resistance to colistin in pigs. We also detail and discuss the One Health approach and propose guidelines for colistin resistance management.

Catching crew members can heavily contaminate their hands with organic material. They can act as mechanical vector and spread diseases between farms. Hand hygiene is an important issue for the industry as a whole and for human health by reducing contamination risks. Many studies, in human medicine, tend to make hand rub a standard for hand hygiene. However, few studies have tested the effectiveness of hand hygiene products on visibly contaminated hands. The objective of this study was to evaluate the effectiveness of practical hand sanitization protocols: water and soap, degreasing cream and hand wipes, all combined with alcohol-based hand gel. The use of alcohol-based gel alone was also evaluated. For the reduction of coliforms after washing, there was no statistically significant difference between protocols when the initial level of bacterial contamination was low to moderate. When hands were highly contaminated, the alcohol-based gel alone was less effective than the degreasing cream combined with the alcohol-based gel (p=0.002). As for the reduction in total aerobic bacteria counts, there was no difference between protocols when the initial level of bacterial contamination was low. The water, soap and alcohol-based gel protocol was more effective than the scrubbing wipes and alcohol-based gel protocol when hands were moderately (p=0.002) and highly contaminated (p=0.001). All protocols were effective in neutralizing Salmonella on hands. Reducing the level of bacterial contamination on hands before using an alcohol-based gel seems important to ensure effective hand sanitation for highly and moderately contaminated hands. This can be done by using a degreasing cream or water and soap. Based on the survey, catching crew members preferred using warm water and soap compared to a degreasing cream.

Aims The object of this study was to determine the impact of only modifying the processing and/or particle size of pig feed on Salmonella shedding and faecal microbiota. Methods and Results Pigs were fed a diet that varied only by their processing (pellet or mash) and their particle size (500, 750 or 1250 μm) for 21 days. Salmonella detection in faeces and seroconversion were determined. Faecal microbiota was assessed by Ion Torrent amplicon sequencing and real-time PCR. Significantly fewer pigs (p

An observational study was conducted of chicken and turkey flocks slaughtered at federal processing plants in the province of Quebec, Canada. The objectives were to estimate prevalence of drug use at hatchery and on farm and to identify antimicrobial resistance (AMR) in cecal Escherichia coli and Enterococcus spp. isolates and factors associated with AMR. Eighty-two chicken flocks and 59 turkey flocks were sampled. At the hatchery, the most used antimicrobial was ceftiofur in chickens (76% of flocks) and spectinomycin in turkeys (42% of flocks). Virginiamycin was the antimicrobial most frequently added to the feed in both chicken and turkey flocks. At least 1 E. coli isolate resistant to third-generation cephalosporins was present in all chicken flocks and in a third of turkey flocks. Resistance to tetracycline, streptomycin, and sulfisoxazole was detected in90% of flocks for E. coli isolates. Antimicrobial resistance (AMR) was observed to bacitracin, erythromycin, lincomycin, quinupristin-dalfopristin, and tetracycline in both chicken and turkey flocks for Enterococcus spp. isolates. No resistance to vancomycin was observed. The use of ceftiofur at hatchery was significantly associated with the proportion of ceftiofur-resistant E. coli isolates in chicken flocks. In turkey flocks, ceftiofur resistance was more frequent when turkeys were placed on litter previously used by chickens. Associations between drug use and resistance were observed with tetracycline (turkey) in E. coli isolates and with bacitracin (chicken and turkey), gentamicin (turkey), and tylosin (chicken) in Enterococcus spp. isolates. Further studies are needed to provide producers and veterinarians with alternative management practices and tools in order to reduce the use of antimicrobial feed additives in poultry.Une étude observationnelle portant sur des élevages de poulets et de dindons abattus dans des usines de transformation sous inspection fédérale situées dans la province de Québec (Canada) a été réalisée. Les objectifs étaient d’estimer la prévalence d’utilisation de médicaments au couvoir et en ferme, la prévalence de résistance aux antimicrobiens dans des isolats caecaux d’

Staphylococcus aureus produces enterotoxins, which are causative agents of foodborne intoxications. Enterotoxins are single-chain polypeptides and have a molecular weight of about 26-28kDa. The consumption of food contaminated with Staphylococcus aureus enterotoxins results in the onset of acute gastroenteritis within 2-6h. The objective of this study was the development of a new method for the quantification of Staphylococcal enterotoxin B (SEB) in food matrices. Tryptic peptide map was generated and nine proteolytic fragments were clearly identified (sequence coverage of 35%). Among these, three specific tryptic peptides were selected to be used as surrogate peptides and internal standards for quantitative analysis using an isotopic tagging strategy along with analysis by LC-MS/MS. The linearity of the measurement by LC-MS/MS was evaluated by combining mixtures of both isotopes at 0.1, 0.2, 0.5, 1.0 and 2.0 1 H/ 2 H molar ratios with a slope near to 1, values of R 2 above 0.98 and %CV obtained from six repeated measurement was below 8%. The precision and accuracy of the method were assessed using SEB spiked in chicken meat homogenate samples. SEB was fortified at 0.2, 1 and 2 pmol/g. The accuracy results indicated that the method can provide accuracy within a 84.9-91.1% range. Overall, the results presented in this manuscript show that proteomics-based methods can be effectively used to detect, confirm and quantify SEB in food matrices. Copyright ©2 011 John Wiley &S ons, Ltd.

Aims: To investigate the effect of continuous feeding of antimicrobial growth promoters (tylosin or virginiamycin) on the swine faecal community. Methods and Results: The study consisted of two separate on-farm feeding trials. Swine were fed rations containing tylosin (44 or 88 mg kg−1 of feed) or virginiamycin (11 or 22 mg kg−1 of feed) continuously over the growing/finishing phases. The temporal impact of continuous antimicrobial feeding on the faecal community was assessed and compared to nondosed control animals through anaerobic cultivation, the analysis of community 16S rRNA gene libraries and faecal volatile fatty acid content. Feeding either antimicrobial had no detectable effect on the faecal community. Conclusions: Erythromycin methylase genes encoding resistance to the macrolide–lincosamide–streptogramin B (MLSB) antimicrobials are present at a high level within the faecal community of intensively raised swine. Continuous antimicrobial feeding over the entire growing/finishing phase had no effect on community erm-methylase gene copy numbers or faecal community structure. Significance and Impact of the Study: Antimicrobial growth promoters are believed to function by altering gut bacterial communities. However, widespread MLSB resistance within the faecal community of intensively raised swine likely negates any potential effects that these antimicrobials might have on altering the faecal community. These findings suggest that if AGP-mediated alterations to gut communities are an important mechanism for growth promotion, it is unlikely that these would be associated with the colonic community.

Salmonella enterica serovar Typhimurium is an important pathogen in swine and is also a frequently reported zoonotic agent. The objective of this study was to characterize isolates of S. enterica serovar Typhimurium associated with septicemia in swine and to compare them to isolates recovered from clinically healthy pigs. We were particularly interested in comparing the two groups of isolates for their ability to adhere to and invade host cells, to be phagocytized and survive in monocyte cells, to induce apoptosis, and to adhere to intestinal mucus. Their surface properties were also evaluated by interactions with solvents. The isolates recovered from diseased animals were shown to invade intestinal epithelial cell lines at a higher rate ( P = 0.003) than isolates from healthy pigs. Septicemic isolates were phagocytized by human monocytes at a higher rate than isolates from healthy pigs ( P = 0.009). The mean percentages of phagocytosis were significantly lower for human monocytes than for porcine monocytes ( P = 0.02 and P = 0.008, respectively) for isolates from both diseased and healthy animals. Healthy animal isolates were phagocytized more by porcine monocytes at 15 min ( P = 0.02) than septicemic isolates. No difference between isolates from septicemic pigs and isolates from healthy pigs was detected for other tested parameters. These results suggest that septicemic isolates have a particular pattern of invasion.

Two experiments were conducted to evaluate lignin and mannanoligosaccharides as alternatives to antibiotic growth promoters in broilers. Dietary treatments for the 2 studies were 1) negative control (CTL-, antibiotic free); 2) positive control (CTL+, diet 1 + 11 mg of virginiamycin/kg); 3) mannanoligosaccharide (MOS; diet 1 + BioMos: 0.2% to 21 d and 0.1% thereafter); 4) LL (diet 1 + 1.25% Alcell lignin); and 5) HL (diet 1 + 2.5% Alcell lignin). In experiment 1, each treatment was assigned to 4 pen replicates (52 birds each). Body weight and feed intake were recorded weekly for 38 d. At 28 and 38 d, cecal contents were assayed for lactobacilli and bifidobacteria. Body weight and feed intake did not differ among dietary treatments. At d 38, the lactobacilli population was greatest (P0.05) in birds fed MOS, whereas LL-fed birds had greater (P0.05) lactobacilli load than those fed CTL+. Bifidobacteria load was greater (P0.05) in birds fed MOS or LL compared with those fed CTL+ at both d 28 and 38. However, at d 28 and 38, lactobacilli and bifidobacteria loads were lowest (P0.05) in CTL+ or HL-fed birds. In experiment 2, 21-d-old birds from the initial flock were transferred to cages for oral Escherichia coli (O2 and O88 serotypes) challenge (12 birds/treatment). After 3, 6, and 9 d, cecal loads of E. coli were determined. Birds fed HL had a lower E. coli load (P0.05) than birds fed CTL- or CTL+ at d 3, and lower than birds fed CTL- at d 6. At d 9, the E. coli load was lower (P0.05) in birds fed MOS or HL than in those fed the CTL- or CTL+ diets; LL-fed birds had lower E. coli load than those fed CTL-. Birds fed MOS or LL had a comparative advantage over CTL+ birds in increasing populations of lactobacilli and bifidobacteria and lowering E. coli loads after challenge.

An observational study was conducted to estimate prevalence and risk factors for carcass contamination by Salmonella and Campylobacter spp. in 60 lots of turkey slaughtered over 10 months in the province of Quebec, Canada. Carcass contamination was evaluated by the carcass rinse technique for about 30 birds per lot. Exposure to potential risk factors was evaluated with questionnaires, meteorological data, and cecal cultures. Multivariable binomial negative regression models were used for risk factor analysis. Prevalence of Salmonella-positive carcasses was 31.2% (95% confidence interval, 22.8 to 39.5%). Variables positively associated (Por = 0.05) with the proportion of lot-positive carcasses wereor =0.5% of carcass condemnation due to various pathologies, cecal samples positive for Salmonella, low wind speed during transportation, closure of lateral curtains of truck during transportation, and slaughtering on a weekday other than Monday. When only Salmonella-positive cecal culture lots were considered, the proportion of carcasses positive for Salmonella was significantly higher in lots exposed to a5 degrees C outside temperature variation during transportation, slaughtered on a weekday other than Monday, and in whichor = 4% of carcasses had visible contamination. Prevalence of Campylobacter-positive carcasses was 36.9% (95% confidence interval, 27.6 to 46.3%). The proportion of positive carcasses was significantly higher in lots with Campylobacter-positive cecal cultures and lots undergoingor =2 h of transit to slaughterhouse. For lots with Campylobacter-positive cecal cultures, variables significantly associated with an increased incidence of carcass contamination were de4% of carcasses with visible contamination, crating foror =8 h before slaughtering, and no antimicrobials used during rearing.

The recent emergence of multidrug-resistant Salmonella enterica subspecies enterica serotype Typhimurium phage-type DT104 represents a challenge to health workers from both human and veterinary medicine. In an effort to develop effective control strategies, it is vital to characterize accurately the extent of genetic and phenotypic variation present in the pathogen population. While previous studies have focused on disease-associated populations, we report the genetic diversity and the diversity in antimicrobial resistance associated with asymptomatic Salmonella Typhimurium DT104 isolates found in pigs. Using pulsed-field gel electrophoresis (XbaI and SpeI), we identified 62 different genotypes associated to 13 different antimicrobial resistant phenotypes among 80 asymptomatic DT104 isolates collected from the Canadian swine industry. When rarefaction curves of a subsample the asymptomatic isolates were compared to that of disease-associated isolates from a similar spatial and temporal scale, we identified significantly more genotypes among the asymptomatic populations with 27 PFGE patterns against 23.0+/-1.4 (95% confidence intervals) in the latter. Also, using Simpson's diversity index, we observed considerably higher genetic and phenotypic diversity in the asymptomatic isolates within herds and detected possible patterns of periodic selection within the disease-associated population. We thus concluded that an important aspect of Salmonella population structure and ecology is overlooked in investigations of disease-associated isolates.

Campylobacter jejuni is an important zoonotic foodborne pathogen causing acute gastroenteritis in humans. Chickens are often colonized at very high numbers by C. jejuni, up to 109 CFU per gram of caecal content, with no detrimental effects on their health. Farm control strategies are being developed to lower the C. jejuni contamination of chicken food products in an effort to reduce human campylobacteriosis incidence. It is believed that intestinal microbiome composition may affect gut colonization by such undesirable bacteria but, although the chicken microbiome is being increasingly characterized, information is lacking on the factors affecting its modulation, especially by foodborne pathogens. This study monitored the effects of C. jejuni chicken caecal colonization on the chicken microbiome in healthy chickens. It also evaluated the capacity of a feed additive to affect caecal bacterial populations and to lower C. jejuni colonization. From day-0, chickens received or not a microencapsulated feed additive and were inoculated or not with C. jejuni at 14 days of age. Fresh caecal content was harvested at 35 days of age. The caecal microbiome was characterized by real time quantitative PCR and Ion Torrent sequencing. We observed that the feed additive lowered C. jejuni caecal count by 0.7 log (p, [À l'origine dans / Was originally part of : Fac. Méd. vétérinaire - Chaire de recherche en salubrité des viandes]

The aim of this study was to examine changes in antimicrobial resistance (AMR) phenotype and virulence and AMR gene profiles in Escherichia coli from pigs receiving in-feed antimicrobial medication following weaning and the effect of feed supplementation with a clay mineral, clinoptilolite, on this dynamic. Eighty E. coli strains isolated from fecal samples of pigs receiving a diet containing chlortetracycline and penicillin, with or without 2% clinoptilolite, were examined for antimicrobial resistance to 15 antimicrobial agents. Overall, an increased resistance to 10 antimicrobials was observed with time. Supplementation with clinoptilolite was associated with an early increase but later decrease in blaCMY-2, in isolates, as shown by DNA probe. Concurrently, a later increase in the frequency of blaCMY-2 and the virulence genes iucD and tsh was observed in the control pig isolates, being significantly greater than in the supplemented pigs at day 28. Our results suggest that, in the long term, supplementation with clinoptilolite could decrease the prevalence of E. coli carrying certain antimicrobial resistance and virulence genes.

Pork meat is estimated to be responsible of 10 to 20% of human salmonellosis cases. Control strategies at the farm could reduce contamination at the slaughterhouse. One of the targeted sector of the production is the maternity, where sows could be Salmonella reservoirs. The aim of this study was to characterize the faecal microbiome of sows excreting or not Salmonella during gestation phases. A total of 76 sows were selected and fecal matters were analysed at the beginning or the end of gestation period. Salmonella detection was conducted using a method including two selective enrichment media (MSRV and TBG). Nine (9) isolates per positive samples were collected. Among the 76 sows tested, 31 were shedding Salmonella. The sows in the first third of their gestation shed significantly more frequently Salmonella (22/29) than those in the last third (9/47) (χ2 P < 0.05). The shedding status of 19 of the sows that were previously sampled in the first third of their gestation was followed, this time in the last third, confirming reduction of the shedding. Association between changes in the intestinal microbiome and this evolution of Salmonella shedding will be explored. MiSeq sequencing is currently being conducted on the feces to identify shifts in the composition or diversity in the microbial community that could be associated to these variations. A large number of Salmonella isolates that were collected were genotyped by a high resolution melt (HRM) technique. These results showed the presence of a major HRM profile (136 isolates / 169) and two minor profiles (24 and 9 /169).