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Salmonella is a foodborne pathogen commonly associated with poultry products. The aims of this work were to (i) estimate the impact of critical steps of the slaughter process on Salmonella detection from broiler chicken carcasses in two commercial poultry slaughter plants in Quebec, Canada; (ii) investigate the presence of Salmonella in the slaughter plant environment; (iii) describe, using a high-resolution melting (HRM) approach, the HRM Salmonella profiles and serotypes present on carcasses and in the slaughter plant environment; and (iv) evaluate whether the HRM flock status after chilling could be predicted by the flock status at previous steps of the slaughter process, the status of previous flocks, or the status of the processing environment, for the same HRM profile. Eight visits were conducted in each slaughter plant over a 6-month period. In total, 379 carcass rinsates from 79 flocks were collected at five critical steps of the slaughter process. Environmental samples were also collected from seven critical sites in each slaughter plant. The bleeding step was the most contaminated, with >92% positive carcasses. A decrease of the contamination along the slaughtering process was noted, with carcasses sampled after dry-air chilling showing ≤2.5% Salmonella prevalence. The most frequently isolated serotypes were Salmonella Heidelberg, Kentucky, and Schwarzengrund. The detection of the Salmonella Heidelberg 1-1-1 HRM profile on carcasses after chilling was significantly associated with its detection at previous steps of the slaughter process and in previously slaughtered flocks from other farms during a same sampling day. Results highlight the importance of the chilling step in the control of Salmonella on broiler chicken carcasses and the need to further describe and compare the competitive advantage of Salmonella serotypes to survive processing. The current study also illustrates the usefulness of HRM typing in investigating Salmonella contamination along the slaughter process. HIGHLIGHTS

The intestinal microbiota plays several important roles in pig health and growth. The aim of the current study was to characterize the changes in the fecal microbiota diversity and composition of weaned piglets following an oral challenge with an ETEC: F4 strain and/or a treatment with colistin sulfate (CS). Twenty-eight piglets were used in this experiment and were divided into four groups: challenged untreated, challenged treated, unchallenged treated, and unchallenged untreated. Rectal swab samples were collected at five sampling times throughout the study. Total genomic DNA was used to assess the fecal microbiota diversity and composition using the V4 region of the 16S rRNA gene. The relative abundance, the composition, and the community structure of piglet fecal microbiota was highly affected by the ETEC: F4 challenge throughout the experiment, while the oral treatment with CS, a narrow spectrum antibiotic, resulted in a significant decrease of E. coli/Shigella populations during the treatment period only. This study was the first to identify some gut microbiota subgroups (e.g., Streptococcus, Lachnospiraceae) that are associated with healthy piglets as compared to ETEC: F4 challenged animals. These key findings might contribute to the development of alternative strategies to reduce the use of antimicrobials in the control of post-weaning diarrhea in pigs.

Toxoplasmosis is a zoonosis of public health concern worldwide. Toxoplasma gondii can be transmitted to humans through the consumption of raw or undercooked meat, but little is known about...

The ever-increasing problem of antibiotic resistance makes routine use of antibiotics in animal production no longer considered as a reasonable and viable practice. The Chicken Farmers of Canada have developed and are implementing an Antimicrobial Use Reduction Strategy, which has the ultimate goal of eliminating the preventive use of medically important antibiotics in broiler chicken and turkey production. However, very little is known about the real overall impact of an antibiotic use reduction strategy in complex ecosystems, such as the bird intestine or the commercial broiler chicken farm. The main objectives of the present study were to compare the abundance of antibiotic resistance-encoding genes, characterize the intestinal microbiota composition, and evaluate the presence of Clostridium perfringens, in six commercial poultry farms adopting short-term antibiotic withdrawal and long-term judicious use strategy. Implementing an antibiotic-free program over a 15-months period did not reduce the abundance of many antibiotic resistance-encoding genes, whereas the judicious use of antibiotics over 6 years was found effective. The short-term antibiotic withdrawal and the long-term judicious use strategy altered the intestinal microbiota composition, with the Ruminococcaceae and Lachnospiraceae families being negatively impacted. These findings are in agreement with the lower production performance and with the increased C. perfringens populations observed for farms phasing out the use of antibiotics. Adopting a conventional rearing program on commercial broiler chicken farms selected for specific antibiotic resistance-encoding genes in many barns. This study highlights the potential impacts of different rearing programs in poultry production and will help guide future policies in order to reduce the use of antibiotics while maintaining production performance.

Tail-biting (TB) in pigs is a serious behavioral disorder. It is an important challenge in swine production as it impacts animal welfare and health and the economics and safety of the pork meat supply chain. To prevent TB, approaches including enrichment material and tail docking are proposed but none are optimal. Nutrition appears to be an important factor in TB behavior, perhaps by modulating the intestinal microbiota (IM). Our aim was to assess the association between TB behavior and IM in pigs through comparisons of IM in groups of biter, bitten and non-biter/non-bitten pigs. Each group composed of 12 pigs was formed at the beginning of the growing/finishing phase based on a target behavior analysis centered on TB behavior for the biter group and a score of damages caused to the tail for the bitten group. Blood and fecal samples were collected from each pig during a TB episode, at time 0, t0, and when the TB episode was considered finished, 4 weeks later, at time 1, t1. Serum cortisol level was determined by ELISA and used as an indicator of stress. The pig's fecal microbiota was analyzed from DNA extracted from freshly collected fecal matter using amplicon sequencing of the V4 hypervariable region of the 16S rRNA gene. Serum cortisol levels were significantly higher in either the biter or bitten pig groups compared to the negative control group (p = 0.02 and p = 0.01, respectively). The microbiota alpha-diversity was not significantly different between all groups, biter, bitten and negative control. Analyses of beta-diversity, however, revealed a significant difference between either the biter or the bitten group in comparison to the non-biter/non-bitten negative control group in terms of structure and composition of the microbiota. Lactobacillus were significantly more abundant in the negative control group compared to the two other groups (p = 0.001). No significant difference was revealed between the biter and bitten groups. Quantitative real-time PCR (qPCR) confirmed that lactobacilli were more abundant in the negative control group. Our study indicates that TB behavior is associated with the IM composition in pigs.

Background: Tail-biting (TB) is a serious behavioral disorder in pigs. It is defined as a pig chewing the tail of another pig. It is an important challenge in swine production as it impacts the animal welfare, its health and the economics and safety of the pork meat supply chain. Some treatments have been proposed but have not proven optimal. Nutrition, feed type and composition, appears to be an important factor in TB behavior, perhaps by modulating the intestinal microbiota (IM). In humans, IM is increasingly recognized as a modulator of behavior. Our aim was to assess an association between TB behavior and IM in pigs through comparisons of IM in groups of biter or bitten pigs to the IM in a non-biter and non-bitten negative control group, and through comparisons of IM between the biter and bitten groups. Each group, composed of 12 pigs, was formed at the beginning of the growing/finishing phase based on a target behavior analysis centered on TB behavior for the biter group and a score of damages caused to the tail for the bitten group. Fecal and blood samples were collected from each pig during a TB episode, at time 0, t0, and after a TB episode, four weeks later, at time 1, t1. Serum cortisol level was determined by ELISA and used as an indicator of stress. The pig’s fecal microbiota was analyzed from DNA extracted from freshly collected fecal matter using amplicon sequencing of the V4 hypervariable region of the 16S rRNA gene. Results: Serum cortisol levels were significantly higher in either the biter or bitten pig groups compared to the negative control group ( p =0.02 and p =0.01, respectively). Interestingly, no significant difference was revealed between the biter and bitten groups. The microbiota alpha-diversity was not significantly different between all groups, biter, bitten and negative control. Analyses of beta-diversity, however, revealed a significant difference between either the biter or the bitten group in comparison to the negative control group in terms of structure and composition of the microbiota. Lactobacillus were significantly more abundant in the negative control group compared to the two other groups ( p =0.001). No significant difference was revealed between the biter and bitten groups. Quantitative real-time PCR (qPCR) confirmed that lactobacilli were more abundant in the negative control group. Conclusions: This study demonstrates that TB behavior is associated to the IM composition in pigs.

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.

Listeria monocytogenes is the etiological agent of listeriosis, a major foodborne disease and an important public health concern. Contamination of meat with L. monocytogenes occurs frequently at the slaughterhouse. Our aims were; 1) to investigate the distribution of L. monocytogenes in the processing areas of four swine slaughterhouses; 2) to describe the diversity of L. monocytogenes strains by pulsed-field gel electrophoresis; 3) to identify persistent L. monocytogenes strains and describe their distribution; 4) to investigate the associations between persistence of strains and their following characteristics: detection in food isolates, detection in human clinical isolates, and the presence of benzalkonium chloride (BAC) resistance genes. Various operation areas within the four swine slaughterhouses were sampled on four occasions. A total of 2496 samples were analyzed, and L. monocytogenes was successfully isolated from 243 samples. The proportion of positive samples ranged from 32 to 58% in each slaughterhouse and from 24 to 68% in each operation area. Fifty-eight different pulsotypes were identified and eight pulsotypes, present in samples collected during 4 visits, were considered persistent. The persistent pulsotypes were significantly more likely to be detected in food (P < 0.01, exact χ²) and human clinical cases (P < 0.01, exact χ²), respectively. Among pulsotypes harboring the BAC bcrABC resistance cassette or the emrE multidrug transporter gene, 42.8% were persistent compared to 4.5% for pulsotypes without these resistance genes (P < 0.01, exact χ²). Our study highlights the importance of persistent L. monocytogenes strains in the environmental contamination of slaughterhouses, which may lead to repeated contamination of meat products. It also shows that the presence of disinfectants resistance genes is an important contributing factor.

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.

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.

Biofilm formation by the pathogen Listeria monocytogenes is a major concern in food industries. The aim of this work was to elucidate the effect of nutrient limitation on both biofilm architecture and on the viability of the bacteria in microfluidic growth conditions. Biofilm formation by two L. monocytogenes strains was performed in a rich medium (BHI) and in a 10-fold diluted BHI (BHI/10) at 30 degrees C for 24 h by using both static conditions and the microfluidic system Bioflux. In dynamic conditions, biofilms grown in rich and poor medium showed significant differences as well in structure and in the resulting biovolume. In BHI/10, biofilm was organized in a knitted network where cells formed long chains, whereas in the rich medium, the observed structure was homogeneous cellular multilayers. Biofilm biovolume production in BHI/10 was significantly higher than in BHI in these dynamic conditions. Interestingly, biovolume of dead cells in biofilms formed under limited nutrient conditions (BHI/10) was significantly higher than in biofilms formed in the BHI medium. In the other hand, in static conditions, biofilm is organized in a multilayer cells and dispersed cells in a rich medium BHI and poor medium BHI/10 respectively. There was significantly more biomass in the rich medium compared to BHI/10 but no difference was noted in the dead/damaged subpopulation showing how L. monocytogenes biofilm could be affected by the growth conditions. This work demonstrated that nutrient concentration affects biofilm structure and the proportion of dead cells in biofilms under microfluidic condition. Our study also showed that limited nutrients play an important role in the structural stability of L. monocytogenes biofilm by enhancing cell death and liberating extracellular DNA.

Extensive use of antibiotic growth promoters (AGPs) in food animals has been questioned due to the globally increasing problem of antibiotic resistance. For the poultry industry, digestive health management following AGP withdrawal in Europe has been a challenge, especially the control of necrotic enteritis. Much research work has focused on gut health in commercial broiler chicken husbandry. Understanding the behavior of Clostridium perfringens in its ecological niche, the poultry barn, is key to a sustainable and cost-effective production in the absence of AGPs. Using polymerase chain reaction and pulsed-field gel electrophoresis, we evaluated how the C. perfringens population evolved in drug-free commercial broiler chicken farms, either healthy or affected with recurring clinical necrotic enteritis outbreaks, over a 14-month period. We show that a high genotypic richness was associated with an increased risk of clinical necrotic enteritis. Also, necrotic enteritis-affected farms had a significant reduction of C. perfringens genotypic richness over time, an increase in the proportion of C. perfringens strains harboring the cpb2 gene, the netB gene, or both. Thus, necrotic enteritis occurrence is correlated with the presence of an initial highly diverse C. perfringens population, increasing the opportunity for the selective sweep of particularly virulent genotypes. Disease outbreaks also appear to largely influence the evolution of this bacterial species in poultry farms over time.

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.

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.

Pathogenic microorganisms in manure represents a potential hazard to human and animal. Few data describing the microbiological content of liquid hog manure intended for spreading are available in the scientific literature. The objective of this project was to verify the presence and estimate the persistence of Escherichia coli , Salmonella spp . , Yersinia enterocolitica and Cryptosporidium spp. in liquid hog manure from storages under conditions typical of commercial swine production in Quebec. E. coli content of liquid hog manure taken during the spreading period in the 32 hog operations varied between 0 and 5.52 log 10 CFU/g with a 95% confidence interval of 3.16–4.23 log 10 CFU/g. Salmonella spp., Y. enterocolitica and Cryptosporidium spp. were detected in 37%, 9% and 3% samples, respectively. Under the conditions of this experiment, the predicted number of days required to obtain a 90% reduction of E. coli populations varied between 15 and 26 days, while the predicted storage period necessary to reach undetectable levels was 54–114 days. The maximal persistence of Salmonella observed in this study was 88 days. A 73-day storage time was insufficient for the elimination of Y. enterocolitica .

The objective of this study was to evaluate the efficiency of a low temperature anaerobic treatment to reduce viable populations of indicator microorganisms (total coliforms, Escherichia coli) and the presence of selected pathogens (Salmonella, Yersinia enterocolitica, Cryptosporidium and Giardia) in swine slurries from different sources. Experiments were carried out in 40 l Sequencing Batch Reactors (SBRs). Experimental results indicated that anaerobic digestion of swine manure slurry at 20 � C for 20 days in an intermittently fed SBR: (1) reduced indigenous populations of total coliforms by 97.94–100%; (2) reduced indigenous populations of E. coli by 99.67–100%; (3) resulted in undetectable levels of indigenous strains of Salmonella, Cryptosporidium, and Giardia .I t can be considered as a promising method for reducing indigenous indicator and pathogenic microorganisms populations in liquid swine manure slurries. � 2005 Elsevier Ltd. All rights reserved.

Background: Campylobacter jejuni is responsible for human foodborne enteritis. This bacterium is a remarkable colonizer of the chicken gut, with some strains outcompeting others for colonization. To better understand this phenomenon, the objective of this study was to extensively characterize the phenotypic performance of C. jejuni chicken strains and associate their gut colonizing ability with specific genes. Results: C. jejuni isolates (n = 45) previously analyzed for the presence of chicken colonization associated genes were further characterized for phenotypic properties influencing colonization: autoagglutination and chemotaxis as well as adhesion to and invasion of primary chicken caecal cells. This allowed strains to be ranked according to their in vitro performance. After their in vitro capacity to outcompete was demonstrated in vivo, strains were then typed by comparative genomic fingerprinting (CGF). In vitro phenotypical properties displayed a linear variability among the tested strains. Strains possessing higher scores for phenotypical properties were able to outcompete others during chicken colonization trials. When the gene content of strains was compared, some were associated with different phenotypical scores and thus with different outcompeting capacities. Use of CGF profiles showed an extensive genetic variability among the studied strains and suggested that the outcompeting capacity is not predictable by CGF profile. Conclusion: This study revealed a wide array of phenotypes present in C. jejuni strains, even though they were all recovered from chicken caecum. Each strain was classified according to its in vitro competitive potential and its capacity to compete for chicken gut colonization was associated with specific genes. This study also exposed the disparity existing between genetic typing and phenotypical behavior of C. jejuni strains., [À l'origine dans / Was originally part of : Fac. Méd. vétérinaire - Chaire de recherche en salubrité des viandes]

Liquid hog manure is routinely applied to farm land as a crop fertilizer. However, this practice raises food safety concerns, especially when manure is used on fruit and vegetable crops. The objectives of this project were to evaluate the persistence of Escherichia coli and Salmonella in surface soil after application of liquid hog manure to fields where pickling cucumbers were grown and to verify the microbiological quality of harvested cucumbers. Mineral fertilizers were replaced by liquid hog manure at various ratios in the production of pickling cucumbers in a 3-year field study. The experimental design was a randomized complete block comprising four replicates in sandy loam (years 1, 2, and 3) and loamy sand (year 3). Soil samples were taken at a depth of 20 cm every 2 weeks after June application of organic and inorganic fertilizers. Vegetable samples were also taken at harvest time. Liquid hog manure, soil, and vegetable (washed and unwashed) samples were analyzed for the presence of Salmonella and E. coli. An exponential decrease of E. coli populations was observed in surface soil after the application of manure. The estimated average time required to reach undetectable concentrations of E. coli in sandy loam varied from 56 to 70 days, whereas the absence of E. coli was estimated at 77 days in loamy sand. The maximal Salmonella persistence in soil was 54 days. E. coli and Salmonella were not detected in any vegetable samples.

Campylobacter spp. were recovered from 660 (77.6%) of 850 swine cecal contents at the abattoir and from 24 (8.6%) of 278 specimens from sporadic cases of human diarrhea during the same period in the same geographical area. Campylobacter coli represented 95.7% of Campylobacter isolates recovered from pigs and 8.3% of those isolated from humans. Genetic profiles were determined by pulsed-field gel electrophoresis (PFGE) using KpnI enzyme to characterize the isolates in combination with phenotypic assays to detect production of cytotoxins, enterotoxins, and hemolysins. Among a subset of isolates (n = 10), up to five colonies from the same animal were characterized by PFGE. In 5 (50%) of 10 of the isolates, more than one genetic profile was observed per pig. Among the 100 isolates from pigs selected for further analysis, 81 different genetic profiles were observed, whereas 20 different genetic profiles were found among the 24 isolates of human origin. Cytotoxicity on Chinese hamster ovary cells was observed in 11 (11%) of 98 isolates from pigs and in 5 (21%) of 24 Campylobacter isolates from humans. No enterotoxin production was detected in Campylobacter isolates in this study, but 17 (71%) of 24 human and 61 (63%) of 97 pig isolates showed hemolytic activity. The study of genotypic and phenotypic profiles of swine and human isolates revealed no epidemiological relationship between isolates. The low genomic relatedness observed between groups of isolates and the weak toxicity level of swine isolates suggest that the hazard of contamination of humans by Campylobacter associated with swine production is low.

Five porcine finishing units, previously identified as contaminated by Salmonella, were sampled to identify possible sources of contamination and to study the distribution of Salmonella within the herds. A total of 208 environmental samples were taken and 87 samples (42%) were found contaminated by Salmonella spp. Salmonella was recovered from several types of samples. Among these, fecal material from pens, building environment such as doors, floors, ventilation units, dust and farm accessories were most often found positive. Some of the flies and rodents were also positive. Two of the finishing units were part of an integrated production system and the prevalence and distribution of Salmonella spp. at different production steps of the integrated facilities were studied. Forty-one farms were sampled and a total of 1923 faecal samples in randomly selected pens were analysed. One hundred and fifty-one samples (7.9%) were positive for Salmonella spp. Among the farms sampled, 70.7% (29/41) were positive for isolation of Salmonella. The different levels in the integrated production were unevenly contaminated. Replacement sow (15.9%) and finishing unit for gilts (21.9%) were the most contaminated levels. Ten serotypes of Salmonella (n = 132) were identified in the production pyramid with a predominance of Salmonella Derby (37.1%) and Salmonella Typhimurium (34.1%). Pulse Field Gel Electrophoresis analysis of the various isolates from serotypes and Salmonella Typhimurium, Salmonella Derby and Salmonella Anatum showed no variation in the genetic profiles, within each serotype, suggesting a vertical contamination throughout the different production steps.

The purpose of this study was to evaluate the risk for human health associated with pathogenic Escherichia coli isolated from airsacculitis and cellulitis in chickens, by comparing the genotypic and phenotypic profiles of avian E. coli isolates and E. coli strains isolated from sick humans during the same period and in the same geographical area as the avian isolates. A total of 96 isolates and 46 isolates from lesions of cellulitis and airsacculitis, respectively, were obtained. Isolates from the backs of some of the affected and healthy birds and 91 intestinal and extraintestinal isolates from humans with diarrhea, urinary tract infections, or septicemia were examined. The frequency of antimicrobial resistance was in general higher in the avian than in the human isolates. VT1-VT2-Eae and VT2-Eae, pathotypes associated with hemolytic and uremic syndrome and bloody diarrhea in humans, were the most frequently encountered pathotypes in human intestinal isolates but were not recovered from the avian isolates. Aero-Pap-TSH and Aero-TSH were the most frequently encountered pathotypes in avian isolates but were rarely observed in human isolates. No avian isolate was of serogroup O157, whereas many human isolates belonged to this O group. O78 and O2 were the most frequently observed O groups in avian isolates but were rarely found in human isolates. Only two avian isolates demonstrated possible relatedness to human isolates based on pulsed-field gel electrophoresis profiles, but they belonged to different pathotypes. Our results suggest that avian isolates recovered from cellulitis and air sacullitis possess very few of the attributes required to cause diseases in humans. It is also concluded that isolates from cellulitis and airsacculitis do not represent a greater hazard than isolates from the back of healthy birds.

Campylobacter jejuni is an important worldwide foodborne pathogen commonly found as a commensal organism in poultry that can reach high numbers within the gut after colonization. Although information regarding some genes involved in colonization is available, little is known about their distribution in strains isolated specifically from chickens and whether there is a linkage between antimicrobial resistance (AMR) and colonization genes. To assess the distribution and relevance of genes associated with chicken colonization and AMR, a C. jejuni microarray was created to detect 254 genes of interest in colonization and AMR including variants. DNA derived from chicken-specific Campylobacter isolates collected in 2003 (n=29) and 2008 (n=28) was hybridized to the microarray and compared. Hybridization results showed variable colonization-associated gene presence. Acquired AMR genes were low in prevalence whereas chemotaxis receptors, arsenic resistance genes, as well as genes from the cell envelope and flagella functional groups were highly variable in their presence. Strains clustered into two groups, each linked to different control strains, 81116 and NCTC11168. Clustering was found to be independent of collection time. We also show that AMR weakly associated with the CJ0628 and arsR genes. Although other studies have implicated numerous genes associated with C. jejuni chicken colonization, our data on chicken-specific isolates suggest the opposite. The enormous variability in presumed colonization gene prevalence in our chicken isolates suggests that many are of lesser importance than previously thought. Alternatively, this also suggests that combinations of genes may be required for natural colonization of chicken intestines.

With the aim to improve growth of weaning piglets and to minimize incidence of intestinal diseases, the effect of a combination of functional foods able to stimulate the development of systemic and mucosal immune system and to modulate bacterial populations in the gut was evaluated. In this study, we assessed the impact of functional foods on the course of Salmonella infection in piglets. Piglets from different litters were weaned at 21 days of age and assigned to 1 of the 4 feed additives (8 litters per treatment) as follow: 1-control (CTRL), 2-antibiotic (ATB), 3-cocktail of functional foods (CFF), 4-bovine colostrum + cocktail of functional foods (COL-CFF). At 49 days of age (day 0), four piglets per litter were orally inoculated with 1X10 8 CFU of Salmonella Typhimurium. A clinical exam was done for each piglet twice a day. Fecal samples were taken to evaluate the Salmonella shedding before and post-infection (days 1, 3, and 7 post-infection). Before challenge and on days 2 and 6 post-infection, blood samples were taken from all piglets to evaluate serum level of prostaglandin E metabolite (PGEM), tumor-necrosis-factor-α (TNF-α) and interleukin-8 (IL-8). At days 3 and 4 post-infection, pigs from ATB group showed no diarrhea compared to pigs from CTRL and CFF groups. At days 4 postinfection, pigs from COL-CFF group showed no diarrhea compared to pigs from CTRL and CFF groups. Regarding fecal excretion at day 1 post-infection, in ATB group, pigs showed lower Salmonella fecal excretion than in CFF group and the lower weight piglets showed a higher fecal excretion than the higher weight piglets. The ATB group pigs showed a lower Salmonella fecal excretion level than pigs from CTRL group at 3 days post-infection. A significant time effect indicated that serum level of PGEM was significantly reduced 2 and 6 days post-infection in comparison to day 0 (before challenge) whereas TNF-α and IL-8 levels were significantly increased after Salmonella challenge.

This study was conducted to determine plasmid colocalization and transferability of both erm(B) and tet(M) genes in Enterococcus faecalis isolates from abattoir pigs in Canada. A total of 124 E. faecalis isolates from cecal contents of abattoir pigs were examined for antibiotic susceptibility. High percentages of resistance to macrolides and tetracyclines were found. Two predominant multiresistance patterns of E. faecalis were examined by PCR and sequencing for the presence of genes encoding antibiotic resistance. Various combinations of antibiotic resistance genes were detected; erm(B) and tet(M) were the most common genes. Plasmid profiling and hybridization revealed that both genes were colocated on a ~9-kb transferable plasmid in six strains with the two predominant multiresistant patterns. Plasmid colocalization and cotransfer of tet(M) and erm(B) genes in porcine E. faecalis isolates indicates that antibiotic coselection and transferability could occur via this single genetic element. To our knowledge, this is the first report on plasmid colocalization and transferability of erm(B) and tet(M) genes in E. faecalis on a mobile genetic element of ~9 kb. Physical linkage between important antibiotic resistance determinants in enterococci is of interest for predicting potential transfer to other bacterial genera.

Objective To compare innate immune responses of peripheral blood leukocytes from healthy and asymptomatic heaves-affected horses. Animals Heaves-affected horses (n = 5-6) and healthy controls (n = 4–5) kept under low dust environments (pasture or shavings and pellets). Methods Blood neutrophil and neutrophil-depleted cell populations were isolated using MACS system. Cells were incubated with or without bacterial products (lipopolysaccharide (LPS), 100 ng/mL and fMLP, 5 ng/mL, 5 h). Cytokine (IL-1β, IL-8, TNF, IL-4, INFγ and IL-10) and receptor (TLR4) mRNA expression was assessed by qPCR. TNF concentration in culture supernatants and serum samples was assessed using equine specific ELISA. Apoptotic rate of resting and stimulated neutrophils was assessed by flow cytometry using AnnexinV and 7-AAD (18 h) and correlated with early pro-inflammatory cytokine expression in the same cells (5 h). Results Stimulation with bacterial-derived products resulted in overexpression of pro-inflammatory cytokines in both neutrophils (IL-1β and TNF) and neutrophil-depleted leukocytes (IL-1β and IL-8) from heaves-affected horses. Neutrophil survival (18 h) was associated with their early TNF expression, but not IL-8. Neutrophil-depleted leukocytes from these horses also had significantly increased basal TNF mRNA levels. Serum TNF concentration was also significantly higher in heaves-affected horses compared to healthy horses kept in similar environment. Conclusions Altered innate immune response to bacterial products is observable ex vivo in peripheral blood leukocytes from asymptomatic heaves-susceptible horses and is associated with high serum TNF concentration. It remains to be determined if this phenomenon is caused by intrinsic differences in innate immune responses or to cellular priming caused by systemic inflammation.

This study was conducted to characterize the antimicrobial resistance determinants and investigate plasmid colocalization of tetracycline and macrolide genes in Enterococcus faecalis and Enterococcus faecium from broiler chicken and turkey flocks in Canada. A total of 387 E. faecalis and E. faecium isolates were recovered from poultry cecal contents from five processing plants. The percentages of resistant E. faecalis and E. faecium isolates, respectively, were 88.1 and 94% to bacitracin, 0 and 0.9% to chloramphenicol, 0.7 and 14.5% to ciprofloxacin, 72.6 and 80.3% to erythromycin, 3.7 and 41% to flavomycin, 9.6 and 4.3% (high-level resistance) to gentamicin, 25.2 and 17.1% (high-level resistance) to kanamycin, 100 and 94% to lincomycin, 0 and 0% to linezolid, 2.6 and 20.5% to nitrofurantoin, 3 and 27.4% to penicillin, 98.5 and 89.7% to quinupristin-dalfopristin, 7 and 12.8% to salinomycin, 46.7 and 38.5% (high-level resistance) to streptomycin, 95.6 and 89.7% to tetracycline, 73 and 75.2% to tylosin, and 0 and 0% to vancomycin. One predominant multidrug-resistant phenotypic pattern was identified in both E. faecalis and E. faecium (bacitracin, erythromycin, lincomycin, quinupristin-dalfopristin, tetracycline, and tylosin). These isolates were further examined by PCR and sequencing for the genes encoding their antimicrobial resistance. Various combinations of vatD, vatE, bcrR, bcrA, bcrB, bcrD, ermB, msrC, linB, tetM, and tetO genes were detected, and ermB, tetM, and bcrB were the most common antimicrobial resistance genes identified. For the first time, plasmid extraction and hybridization revealed colocalization of tetO and ermB genes on a ca. 11-kb plasmid in E. faecalis isolates, and filter mating experiments demonstrated its transferability. Results indicate that the intestinal enterococci of healthy poultry, which can contaminate poultry meat at slaughter, could be a reservoir for quinupristin-dalfopristin, bacitracin, tetracycline, and macrolide resistance genes.

Despite the application of hazard analysis and critical control point systems at slaughter and during processing, Salmonella contamination is still a significant biological hazard associated with pork products. A better understanding of risk factors in slaughterhouses and of contamination sources is therefore critical to improve control of this bacterium in the abattoirs. The objectives of this study were to identify the risk factors at slaughter that are associated with the presence of Salmonella on hog carcasses and to assess possible sources of contamination. A questionnaire on potential risk factors was developed. Over 7,400 hogs originating from 312 randomly selected production lots were tested. The lots were from 10 different abattoirs located in five different Canadian provinces. At slaughter, blood was collected for serological analysis, and mesenteric lymph nodes (MLN) and carcass swabs were collected for Salmonella analysis. Furthermore, pulsed-field gel electrophoresis was conducted to establish the genetic profiles of selected isolates from carcasses and MLN and to compare these profiles with those recovered from the slaughter environment. Multivariate regression analysis results indicated that the cleanliness of the hogs and the status of the scald water were factors significantly associated with the Salmonella status of the carcasses at the end of the slaughter process. Pulsed-field gel electrophoresis analysis showed that most isolates from carcasses were similar to those from animals (MLN) or the preevisceration environment.

An observational study was conducted to estimate prevalence and risk factors for Salmonella and Campylobacter spp. carcass contamination in broiler chickens. Eighty-two lots were sampled in four slaughterhouses located in the province of Québec, Canada, over a 10-month period. Carcass contamination was evaluated by the carcass rinse technique for about 30 birds per lot. Exposure to potential risk factors was evaluated based on data from questionnaires, meteorology, and cecal cultures. Multivariable binomial negative regression models were used for risk factor analysis at the lot level. The prevalence of Salmonella-positive carcasses was 21.2% (95% confidence interval: 15.7 to 26.7%). Significant risk factors (P0.05) associated with a higher proportion of positive carcasses within lots were Salmonella-positive cecal culture, low rainfall during transportation to the slaughterhouse, temperature ofor = 0 degree C during transportation to the slaughterhouse, and aor = 4-h waiting period in shipping crates before slaughtering. The prevalence of Campylobacter-positive carcasses was 35.8% (95% confidence interval: 27.1 to 44.5%). Lots containing birds with Campylobacter-positive cecal culture results, lots of birds that were slaughtered at the end of the week, and lots with at least 20% of birds with digestive contents detected in the jejunum at time of slaughtering had a significantly higher proportion (P0.05) of contaminated carcasses. These results support the importance of preharvest control measures implemented during rearing to reduce contamination of the final product. Weather during transportation to slaughter and the day of the week that birds were slaughtered also were associated with carcass contamination; further studies are needed to determine the underlying mechanisms by which these factors influence carcass contamination.

We conducted an observational study to estimate prevalence and risk factors for Salmonella spp. and Campylobacter spp. caecal colonization in poultry. Eighty-one broiler chicken and 59 turkey flocks selected among flocks slaughtered in the province of Quebec, Canada, were included in the study. Flock status was evaluated by culturing pooled caecal contents from about 30 birds per flock. Exposure to potential risk factors was evaluated with a questionnaire. Odds ratios were computed using multivariable logistic regression. The prevalence of Salmonella-positive flocks was 50% (95% CI: 37, 64) for chickens and 54% (95% CI: 39, 70) for turkeys, respectively. Odds of Salmonella colonization were 2.6 times greater for chicken flocks which failed to lock the chicken house permanently. In turkeys, odds of Salmonella colonization were 4.8-7.7 times greater for flocks which failed to be raised byor=2 producers with no other visitors allowed onto the premises, or origin from a hatchery. The prevalence of Campylobacter-positive flocks was 35% (95% CI: 22, 49) for chickens and 46% (95% CI: 30, 62) for turkeys. Odds of colonization were 4.1 times higher for chicken flocks raised on farms with professional rodent control and 5.2 times higher for flocks with manure heap200m from the poultry house, and also increased with the number of birds raised per year on the farm and with the age at slaughter. For turkeys, odds of Campylobacter flock colonization were 3.2 times greater in flocks having a manure heap at/=200m from poultry house and 4.2 times greater in flocks drinking unchlorinated water.

Carriers of Salmonella in finishing pigs are believed to be a main source of carcass and pork contamination at the slaughterhouse and during meat processing. To better control the presence of Salmonella at the farm and slaughter, it is important to adequately identify on-farm risk factors associated with presence of these bacteria in animals. Thus a study was performed to identify the risk factors associated with the presence of Salmonella in live animals and on carcasses from 312 herds in Canada. A questionnaire was designed to gather information on several factors present on the farm and was completed by the producers. Results from the multivariate logistic regression analyses of all the factors indicated a significant effect of the type of feed grind (p = 0.0001), pen size (p=0.006), truck load capacity (p=0.008), clinical signs of salmonellosis (p=0.03) and a marginal effect of number of sources of finishing pigs (p = 0.098). Carcasses from highly contaminated herds were more likely of being Salmonella positive. Limiting the presence of Salmonella in the meat should therefore begin by reducing the number of herds highly contaminated by Salmonella. Introduction Many HACCP-based good production practices to be used on farm have been developed in order to manage biological hazards, such as Salmonella and chemical hazards. The purpose of these practices is, ensuring a better use of antimicrobial agents and decrease risk of drug residues in meat products. So far, in pigs, these models have been built based on principles of risk analysis to manage well known risk factors or to avoid various sources that contaminate the herd. Very limited information was available in Canada on risk factors associated with the presence of Salmonella in pig operations. In most cases, we have had to rely on published literature to build on farm control programs. Use of antimicrobial agents in intensive production units has been associated with the emergence of bacterial strains resistant to several antibiotics. While the majority of authors agree that the use of antibiotics in animals can be linked to only a small percentage of the resistances observed among humans, almost all stakeholders agree that antibiotics must be used more judiciously in livestock production in order to stem the emergence of resistant strains, even if only for the sake of animal health. Resistance to multiple antibiotics is regularly observed in Salmonella strains isolated in Canada (Rheault et al., 2001). Furthermore, these multiresistant strains are associated with acute health problems on a growing number of hog farms in Canada (Desrosiers 1999). It has been reported in Europe that use of broad-spectrum antimicrobial agents can foster the presence of Salmonella in livestock (Evans and Wegener, 2003). Since resistant Salmonella isolates have genes encoding resistance to many of the antibiotics commonly used in livestock production, thus ensuring their selection and proliferation, it is important to determine if the antibiotics most commonly used on farms in Canada can be associated with the presence of multiresistant strains. The objectives of this study were to identify the risk factors associated with the presence of Salmonella in swine herds and determine if the presence of multiresistant strains of Salmonella can be associated with the use of some antimicrobial agents on farm. Materials and Methods Questionnaire: A questionnaire was designed to collect information on potential risk factors and was completed by the producers. It addressed risk factors at the farm and to a lesser extent those associated with the transportation of animals. There were three phases in the development of the questionnaire. The first involved reviewing of the literature in order to collate all the risk factors associated with husbandry and transportation. In the second phase, a group of experts in the epidemiology and control of Salmonella in pigs were consulted and asked to validate the documented risk factors listed in the first draft of the questionnaire. They also suggested other potenO R A L P R E S E N T A T IO N S 17 SafePork 2005 tial risk factors. Finally, twenty hog farmers were asked to validate the questionnaire for understanding clarity. In addition, all the participating producers were contacted and given an explanation of the scope and goals of the study, along with instructions on how to complete the questionnaire properly. Most producers were again contacted after they had received the questionnaire to ensure validity of some of their answers. Collection of samples at the slaughterhouse: blood samples, one-gram sample of mesenteric lymph nodes were collected. In addition, bacterial carcass samples were collected by swabbing the three predeterminated anatomical sites were obtained for each animal. A total of 7441 carcasses were included. These pigs originated from 312 production batches and at 10 slaughterhouses in Canada receiving animals from Quebec, Ontario, Manitoba, Saskatchewan and British Columbia. Between 20 to 25 animals were randomly sampled per lot (first animal randomly selected, and every fourth one thereafter). A total of over 22,000 samples were obtained. Bacteria isolation and Characterization: Swabs were placed in sterile bags containing a transportation medium, put in a cool place, and shipped to the laboratory of the Research Chair in Meat Safety of the University of Montreal. The samples were incubated according to the official procedures described in the Mega-Reg (USDA, 1996), in selective enrichment broth (RV and TBG) and then inoculated on selective agar media (BGS and DMLIA supplemented with novobiocin 20μg/mL). Biochemical assays were conducted to confirm the identification, and serotyping was done at the MAPAQ laboratories in Saint-Hyacinthe, Quebec. A total of 432 Salmonella-positive isolates were characterized for their resistance to antibiotics by placing antibiotic discs on the agar media using the NCCLS-approved method. Serological Analysis: Blood samples were analyzed in order to detect the presence of Salmonella antibodies by the means of an ELISA to establish and determine the status of the animals when at farm (Cote et al., 2004). Statistical Analyses: a univariate logistic regression was done for each type of analysis in order to identify significant discrete-type risk factors (presence/absence) in relation with a discrete dependent variables, namely serology and lymph node scores. The serology score was 0:0% seropositive ; 1 = : > 0% and 20% seropositive, where 20% represented the 75th percentile of distribution. The codes for lymph nodes were: 0: 0% positive lymph nodes; 1: > 0% and 74% positive lymph nodes, where 74% represented the 75th percentile of distribution. A multivariate logistic regression was used to establish which of the variables identified in the univariate analysis were the most significant. All these analyses were done using version 8.1 of the SAS program (SAS Institute, Cary, NC). Other analyses were done using Student’s t-tests for comparing prevalence in small populations , unless otherwise indicated. Results The prevalence of seropositive animals, which indicates the status that had the animal several days prior to slaughter was 13.6%. The prevalence observed in lymph nodes, which determines the status of the animal at its entry into the slaughter chain, was 32.3%. The multivariate analysis of risk factors for the seroprevalence of Salmonella revealed a significant effect of the type of feed grind (p = 0.013), pen size (p=0.006), truck load capacity (p=0.008), signs of salmonellosis (p=0.03) and a marginal effect of number of sources of finishing pigs (p = 0.098). The odds of having a high score in serology were nearly 8 times lower (odds ratio = 7.56) with feed in mash form than with feed in pelletted form. The odds of having a high score in serology were lower when the truck load capacity was under 200 pigs (OR=0.32) and when pen size was smaller (under 170 square feet, OR=0.31). The odds were lower without signs of salmonellosis in the herd (OR=0.089). Finally, the odds were lower with one animal source than with two or more sources (OR=0.31). We studied the relationship between the status of the lymph nodes and the serology. In many cases (69/84), the serology was negative whereas the lymph nodes were positive, suggesting that several animals become contaminated at the end of the finishing period or during transportation or the pre-slaughtering period. When the serology was positive, the lymph nodes were often positive as well (86/95), indicating that contamination occurred most likely in the herd at least 3 weeks earlier, which is the length of time usually required to develop a serological response to the infection. At the batch level, results from the logistic regression model, with the sampled slaughterhouse as the random factor, indicated a positive and significant relationship between the % of

Streptococcus suis, an important swine pathogen worldwide, is made up of 35 serotypes (1 to 34 and 1/2) (12). Although all serotypes can cause infections, serotype 2 is the most prevalent one isolated from diseased pigs (9, 11). Septicemia, arthritis, meningitis, and sudden death are the most important clinical signs associated with S. suis infections (11). Cases of meningitis and endocarditis caused by S. suis have also been reported in workers associated with the pig industry (11). Although several S. suis virulence factors have been identified and characterized, the exact mechanisms by which this bacterium invades the host and causes infections are still unclear. The only factor considered essential for the pathogenicity of S. suis is the polysaccharide capsule (3). An important feature, which could play an important role in the pathogenicity of S. suis, is its ability to bind host proteins, which may camouflage it and thus protect it from the host immune system. Fibronectin (7), albumin (21), and immunoglobulin G (1, 23) receptors have been identified on the cell surface of S. suis and have been proposed as virulence factors. Plasminogen is a 92-kDa protein that is an important component of the fibrinolytic system. Its activation into plasmin, a serine protease, is tightly regulated by the equilibrium between plasmin activators (urokinase plasmin activator [u-PA] as well as tissue plasminogen activator) and inhibitors (α2-antiplasmin and α2-macroglobulin) (15). Plasminogen can also be activated by microbial products including streptokinase and staphylokinase produced by group A streptococci and Staphylococcus aureus, respectively (15). Many group A and C streptococci bind plasminogen on their cell surfaces (15). In a number of cases, the receptors have been characterized and identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or α-enolase (17, 19, 20, 24). The plasmin activity generated on the bacterial cell surface is protected from host inhibitors and can activate latent u-PA and tissue plasminogen activator, which subsequently activate plasminogen (15). This mechanism of acquisition of a host proteinase activity increases the invasive potential of some pathogens, including Streptococcus pneumoniae (8). The purpose of the study reported here was to investigate the capacity of S. suis to bind plasminogen and acquire plasmin activity.

The in vitro virulence properties of 197 temporally and geographically related Campylobacter isolates from chicken broilers and humans were compared. Comparisons of the virulence properties associated with genotypes and biotypes were made. All isolates adhered to, and 63% invaded, INT-407 cells, whereas 13% were cytotoxic for CHO cells. CHO cell-cytotoxic extracts were also cytotoxic for INT-407 cells, but the sensitivity for Vero cells was variable. The proportion of isolates demonstrating a high invasiveness potential (>1,000 CFU ml −1 ) or Vero cell cytotoxicity was significantly higher for human than for poultry isolates. Invasiveness was associated with Campylobacter jejuni isolates of biotypes 1 and 2, whereas CHO and INT-407 cell cytotoxicity was associated with C. jejuni isolates of biotypes 3 and 4. Cytotoxic isolates were also clustered according to pulsed-field gel electrophoresis profiles.

Streptococcus suis serotype 2 is a swine pathogen responsible for diverse diseases and may be present in the tonsils of pigs which show no sign of illness. Because adhesion to host cells may be important in the carrier state, this study was undertaken to characterize a 39 kDa surface protein identified as a glyceraldehyde-3-phosphate dehydrogenase (GAPDH), possibly implicated in the adhesion of the bacteria. The gene encoding for the GAPDH of S. suis was cloned and sequenced. The DNA sequence contained an open reading frame encoding for a 336 amino acid polypeptide exhibiting 95% sequence identity with the GAPDH from Streptococcus pyogenes and from other streptococci. Using the Qiaexpress expression plasmids, the gapdh gene was inducibly overexpressed in E. coli to produce GAPDH with a hexahistidyl N-terminus to permit its purification. The (His)6GAPDH protein was found to possess functional GAPDH enzymatic activity after the purification. An adherence assay with S. suis and porcine tracheal rings pre-incubated with (His)6GAPDH and non-incubated rings was showed a significant reduction in the adhesion of S. suis in the (His)6GAPDH pre-incubated rings compared to the non-incubated rings. The GAPDH protein of S. suis seems to be involved in the first steps of the bacterial adhesion to host cells.

C. coli from slaughtered pigs and from human patients were examined for resistance to quinolones and tetracycline. Detection of tetO was performed. Gyrase A gene (gyrA) was amplified and sequenced and tested by an alternative method. Tetracycline resistance levels were respectively of 67,7 % and 56,3 % in swine and human isolates. In C. coli of human origin, all resistant isolates had the tetO determinant while 82,8 % of resistant C. coli from swine possessed it. Among the susceptible swine isolates, 17,2 % possessed this gene. Resistance to enrofloxacin (7,3 %) and ciprofloxacin (11,4 %) was observed in swine isolates and resistance for enrofloxacin (12,5 %) and ciprofloxacin (18,8 %) were observed in C. coli from humans. In addition, 72,7 % of swine resistant isolates and all isolates from humans had a mutation at position 86. Results were similar with MAMA-PCR which can thus be considered as a good alternative to sequencing.

ORAL PRESENTATIONS implications. First, it implies that there is distinction in subtypes between the two hosts. This may further imply that other sources may also be important. Recently, report by Barber et al. stressed the underestimation of the role of non-food animal sources such as humans and pets (Barber et al., 2003). Second, the finding may indicate that only certain subtypes of this serovar are able to cause clinical illness strengthening the notion that not all strains of Salmonella serovar Typhimurium are capable of causing clinical illnesses.

ORAL PRESENTATIONS 3 18 h to eliminate Salmonella or E. coli from feed at 20 oC compared with 30 min – 2 h at 30 C and less than 30 min at 37 C. Lactic acid concentration also plays an important role the elimination of enteropathogens from FLF and in this study high concentrations of lactic acid were achieved via inoculation with Lb plantarum. With proper management, i.e. temperature control and inoculation with lactic acid bacteria to ensure adequate lactic acid production, FLF systems have the potential to play an important role in the reduction of enteropathogens in the food chain.

ORAL PRESENTATIONS negative status for Salmonella, we were interested in this study to determine if it was possible to develop a protocol that would allow the population of a negative farrowing unit by use of animals from a herd known to be moderately contaminated by Salmonella. Since serology can not detect recently infected animals and since bacteriology can not detect most healthy carriers, we used a combination of serology and bacteriology, ensuring that the last sampling was done after the stress period caused by the selection process. In the first part of this study, we observed that all sampled animals from NP/NS pens were negative after introduction in the new farrowing unit. Interestingly, it was also possible to populate an almost negative section of the new unit by use of PP/NS animals. To do so, few basic measures were applied to these animals such as a Neomycin treatment in water and the washing of gilts at their arrival to the new unit. In addition, few special precautions such as maintenance of the integrity of pens and biosecurity measures such as changing/washing boots between positive and negative pens, were applied throughout the experiment.

Between July 1998 and June 1999, 93 lots of broiler chickens distributed on 57 farms were sampled in two abattoirs of the province of Quebec (Canada). A total of 2,325 samples of cecal material were analyzed to determine the prevalence of campylobacters. Biotyping and pulsed-field gel electrophoresis (PFGE) were done on 20% of the Campylobacter isolates to study the distribution within poultry production. Macrorestriction profiles were compared with profiles of 24 Campylobacter strains isolated from sporadic cases of human diarrheic patients in order to evaluate genetic relationships. Approximately 40% of the broiler chickens in 60% of the lots and 67% of the farms were colonized. Biotypes I and II of Campylobacter jejuni were the most prevalent biotypes in poultry and human isolates. The PFGE dendograms revealed a high genetic diversity among poultry isolates, with 49 different genotypes from the 56 positive lots. More than 75% of these lots were colonized by a unique genotype. All positive lots raised simultaneously on the same farm had common genotype(s). Different genotypes were isolated from lots raised at different grow-out periods on a farm. In some cases, identical genotypes were found at different grow-out periods on a farm and also from different farms. Macrorestriction profiles showed that approximately 20% of human Campylobacter isolates were genetically related to genotypes found in poultry. This genetic relationship and the high prevalence of C. jejuni biotypes I and II in poultry indicated that Campylobacter in broiler production of the province of Quebec could be a potential source of hazard for public health.