No abstract

-Although poultry is recognized as the major source of food-poisoning caused by Salmonella, pork also contributes to human infections. This study was therefore undertaken in order to develop a reliable serological method for the evaluation of the Salmonella status of piglets. A complete ELISA was performed using lipopolysaccharides of Salmonella Typhimurium, Anatum, Hadar and Infantis because these serovars were representative of the serogroups isolated from 30 contaminated fattening farms. S. Enteritidis was also added because of its importance in human infection and to include the O : 9 antigen. This method potentially detects 100% of infected pigs. A significant correlation was found between this serological method and the bacteriological data from mesenteric lymph nodes (p = 0.01). In addition, both sensitivity and specificity were high (97% and 94% respectively). The ELISA test was therefore used in a cross-sectional study on 4 farms to evaluate when pigs became contaminated: seropositive pigs were only found for the 20 week old finishing pigs. The antibody response to Salmonella in piglets was also investigated: maternal antibodies persisted until 7 weeks of age and post-Salmonella contamination seroconversion was detected from 8 weeks of age onwards.ELISA / Salmonella / pig / epidemiology Résumé -Mise au point d'une méthode ELISA complète basée sur des lipopolysaccharides de plusieurs sérogroupes permettant de détecter tous les porcs infectés par des salmonelles. Bien que la volaille soit la principale source d'intoxication alimentaire par les salmonelles, le porc contribue aussi à la contamination humaine. Cette étude a été menée pour mettre au point une technique sérologique permettant d'évaluer le statut infectieux des porcelets vis-à-vis des salmonelles. Une technique ELISA dite complète a été élaborée avec des lipopolysaccharides de Salmonella Typhimurium, Anatum, Hadar et Infantis, sérovars représentatifs des sérogroupes isolés dans 30 élevages engraisseurs contaminés. S. Enteritidis a été ajoutée en raison de son importance dans les infections humaines et afin d'inclure l'antigène O : 9. Cette méthode permet de dépister potentiellement 100 % Vet. Res. 31 (2000) [481][482][483][484][485][486][487][488][489][490] 481

-The aim of this study is to assess the risk of contamination by Salmonella Typhimurium of pigs by nose-to-nose contact or the airborne route. Thirty twelve-week-old SPF pigs were divided into 4 groups housed in 4 different rooms: the first room contained Salmonella-free control pigs (n = 4), the second room had 10 3 CFU S. Typhimurium inoculated pigs (n = 5) and non-inoculated "contact" pigs (n = 4), the third room had pigs (n = 8) receiving potentially contaminated air from the following room through a hole (4 pigs housed in the pen situated near the hole and 4 pigs in the pen at the opposite side of the room), and the fourth room had pigs (n = 5) inoculated with 10 6 CFU Salmonella Typhimurium and also non inoculated "contact" pigs (n = 4). The "contact" and the inoculated pigs were housed in adjacent pens allowing nose-to-nose contact. The 5 pigs orally inoculated with 10 6 CFU S. Typhimurium were bacteriologically and serologically positive 1 week later and their environment was contaminated as early as 1 day pi. The faecal samples of 4 nose-to-nose contact pigs were bacteriologically positive and one of them was seropositive 5 weeks pi before the pigs were commingled. The 8 pigs housed in the third room received S. Typhimurium by an active airflow coming from the contaminated room (1000 m 3 /hour). Their faecal samples remained negative until 8 weeks pi but the environmental swabs taken in the room close to the airinlet were contaminated 2 days pi and positive swabs were found elsewhere in the room 5 weeks pi. Two seropositive pigs were encountered 8 weeks pi in the pen situated near the hole. Only one among the 5 pigs inoculated with 10 3 CFU had bacteriologically positive faeces 1-week pi and the 4 pigs kept in noseto-nose contact with them remained negative. A dose of 10 3 CFU was too small to induce persistent excretion and to stimulate a humoral immune response. However, the dose of 10 6 CFU induced contamination of nose-to-nose contact pigs and contamination of the environment by airflow.ELISA / Salmonella / pig / airborne transmission Résumé -Contamination des porcs par contact nez-à-nez ou par voie aérienne par Salmonella Typhimurium. L'objectif de cette étude est d'évaluer le risque de contamination des porcs par contact nez-à-nez et par la voie aérienne. Trente porcs exempts d'organismes pathogènes spéci-fiques âgés de 12 semaines ont été subdivisés en 4 groupes hébergés dans 4 salles différentes : la Vet. Res. 32 (2001) 591-600 591

The heritability of resistance of poultry to Salmonella enteritidis (SE) was investigated. Three m easurem ents of resistance were m ade: survival after intram uscular inoculation of 419 day-ol d chicks, absence versus presence of Salm onella in spleens and caeca 4 weeks after oral inoculation of 304 hens at peak of laying, and antibody response of 228 hens following two inoculations of an aroA m utant of this serotype. In the ® rst two models of infection, resistance appeared to be heritable. The heritability was estim ated from the sire and dam com ponents, respectively, at 0.14 6 0.10 and 0.62 6 0.16 for chick m ortality, 0.47 6 0.21 and 0.13 6 0.26 for resistance to spleen contam ination, and 0.24 6 0.15 and 0.53 6 0.26 for resistance to caecal contam ination in laying hens. By contrast the estim ated heritability of antibody response was very low (0.03 6 0.08 and 0.10 6 0.08 when estim ated from the sire and dam com ponents, respectively). These results suggest that a selection for increased resistance to SE m ay be ef® cient.

-We did a prospective observational 9-month long study to quantify risk factors of managerial and hygiene practices, and pig-health status for Salmonella seroconversion of fattening pigs reared in subclinically infected French farrow-to-finish farms. During the fattening phase, 2 649 pigs belonging to the same batch of contemporary pigs, from 89 conventional farrow-to-finish farms were individually followed and regularly blood sampled on a monthly basis. Farm recruitment was based on the farmer's willingness to cooperate. Pig status was assessed using an indirect ELISA test. Evolution of the serological status was studied by means of survival analysis. A Cox proportionalhazards model, taking into account the clustering of animals at the farm level, was used to examine the effects of explanatory variables on the time to Salmonella seroconversion of pigs. Applying group level antibiotic treatment to the pigs during the fattening period (Hazard Ratio (HR) = 2.4; 95% CI: 1.7, 3.4) was identified as a risk factor for Salmonella seroconversion, as the presence of residual Salmonella contamination in the fattening pen before placing the pigs into the pens (HR = 1.9; 95% CI: 1.2, 2.9). Porcine reproductive and respiratory syndrome virus (PRRSV) seropositivity during the fattening period also indicated an increased hazard for seroconversion (HR = 1.6; 95% CI: 1.1, 2.5). The batch size was identified as a risk factor for Salmonella seroconversion: the higher the number of pigs was in the fattening room followed, the higher was the risk (HR +10pigs = 1.05 for a 10-pig increment; 95% CI: 1.03, 1.06). The biosecurity measures of wearing specific clothes before entering the facilities (HR = 0.5; 95% CI: 0.3, 0.9) and enclosing the pig farm facilities were protective (HR = 0.4; 95% CI: 0.2, 0.8).Salmonella / pigs / seroconversion / risk factors / survival analysis

-The aim of this study was to evaluate the impact of the pre-slaughter process on Salmonella caecal contamination of pigs at slaughter. An observational study was carried out in 2001 on 101 conventional farrow-to-finish pig farms. On each farm, one batch of contemporary pigs was followed from the end of the fattening period until slaughter. The Salmonella bacteriological status of the batches was assessed by environmental samples of faecal material. The serological Salmonella status was obtained on 30 individually identified market-age pigs using an indirect ELISA test. At the slaughterhouse, 25 g of caecal contents were taken from 10 of the identified pigs. Faecal and caecal material were analysed according to a classical bacteriological method. A questionnaire was designed to obtain information about the type of feeding during the fattening period (dry versus wet), the duration of fasting on the farm before leaving for the slaughterhouse, the duration of transport between the farm and the slaughterhouse, the holding time in lairage at the slaughterhouse and loading and unloading conditions on the farm and at the slaughterhouse. To assess the relationships between these factors and the Salmonella caecal status of the pigs and the batches, two logistic models were fitted at the individual and at the batch level, respectively. The first analysis was performed using a random effects logistic regression model. The second analysis was based on a cumulative logit model with a positive caecal rate classified into three classes as the outcome variable. The results showed that the Salmonella status of market-age pigs assessed on the farm either by serological or bacteriological examinations and the time spent in lairage before slaughtering played a crucial role on caecal contamination. In the light of these results, actions should be considered both on the farm and at the slaughterhouse to decrease the risk of Salmonella contamination of the caecal contents.

In France, the regular and compulsory detection of Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) in flocks of breeding and laying hens is based on bacteriological examination of environmental swabs and faeces samples. The aim of this study was to compare this bacteriological examination with a serological method (ELISA) developed in our laboratory. This ELISA was first evaluated by use of artificially infected hens. During these experimental infection studies, several groups of hens were inoculated with SE, ST, different vaccines and different Salmonella serovars to calculate the experimental parameters of our ELISA. Then, in a field study, 43 flocks were followed monthly using two bacteriological samples (environmental swab and pool of faeces) and 20 serological samples (sera or yolks). Twenty-seven flocks without SE or ST gave a negative serological response throughout their surveillance. Among the 10 various serovars different from SE and ST isolated in this study, S. Heidelberg, S. Agona and S. Hadar gave seropositive results in seven flocks. Consequently, this ELISA was not specific of SE and ST as it detected serovars sharing or not common antigens with SE and ST. Seropositive results were also obtained each month for two flocks where no Salmonella could be isolated. Finally, in seven flocks found infected with SE or ST, the positive ELISA results appeared later than the bacteriological detection. Therefore, for the detection of chicken flocks recently infected with SE or ST, bacteriological examination currently used in France seems to be more appropriate than this ELISA.

-Reliable ELISAs were investigated with the aim to select hen lines resistant to Salmonella Enteritidis and producing high levels of antibodies. In the first experiment, the relation between the humoral response and the bacteriological results was assessed on hens from the Y11 resistant line and the L2 susceptible line, orally inoculated with 10 8 CFU S. Enteritidis per animal. Antilipopolysaccharide (LPS) IgG titres were higher but the liver and spleen were less contaminated in hens from the Y11 line than in hens from the L2 line (p = 0.013, 0.031 and 0.026 respectively). In the second experiment, the hens were inoculated orally with 1.7 × 10 8 CFU S. Enteritidis per animal in order to select the ELISA methods showing the more significant differences. ELISAs were based on LPS, flagella, LPS from rough (LPS-R) and smooth strains (LPS-S) and detected IgG and IgM antibodies from sera and yolks. No between-line host response variation was observed in the yolk, with LPS-S and R antigens nor with anti-LPS IgM in the sera. Otherwise, significant differences were encountered between hen lines with the ELISAs performed on the sera detecting anti-LPS IgG, anti-flagella IgG or IgM (p = 0.017, 0.017 and p < 0.001 respectively). When comparing the kinetics of the selected ELISAs, the IgG antibodies against LPS detected between-line variations as early as 1 to 4 weeks pi, whereas with IgG against flagella, the differences were only detected at 1 and 2 weeks pi and with IgM against flagella, the differences were significant at 1, 2, 4 and 8 weeks pi. In conclusion, resistant hen lines producing higher levels of antibodies than the susceptible hen lines may be selected with these ELISAs.