An integrated approach of genetic resistance to Salmonella carrier state in fowls: from genetics to genomics and modelling.

Increasing resistance to acute Salmonellosis (that is, contamination level shortly after infection) is not sufficient to reduce the risk for consumers to be contaminated by Salmonella. Indeed, animals may remain contaminated at a low level for weeks or months. Increased resistance to the Salmonella carrier state, i.e., animals' ability to clear bacteria, is needed; it involves measuring bacterial contamination several weeks after inoculation with a low dose. To study such resistance traits, three convergent approaches were used. A quantitative trait loci (QTL) study was performed, taking advantage of inbred lines differing in resistance. Several QTLs controlling resistance at a younger age were identified and are currently being confirmed in a new cross before finer mapping, using advanced intercross lines. These inbred lines are also presently being compared using functional genomics. In parallel, a selection experiment for increased or decreased resistance at a younger and a later age was undertaken. Besides providing genetic models differing in their levels of resistance, it underlined the importance of the choice of selection criterion, whether marker assisted or not. Indeed, genes controlling resistance are strongly dependant on age; selecting for resistance at a younger age might result in increased susceptibility at an older age. Finally, the results of this experiment were used in a model of the intra-flock propagation of Salmonella. It showed that introducing a proportion of resistant animals within a flock of susceptible hens could dramatically change the evolution of contamination. Moreover, it demonstrated the magnitude of synergy between selection and vaccination, which should enhance the interest of increased resistance. The results show that selection for increased resistance to the Salmonella carrier state may be efficient, providing that the appropriate criteria of selection are used.

Effect of two candidate genes on the Salmonella carrier state in fowl.

Selection for increased resistance to Salmonella carrier-state (defined as the persistency of the bacteria 4 wk after inoculation) could reduce the risk for the consumer of food toxi-infections. The effects of two genomic regions on chromosomes 7 and 17 harboring two genes, NRAMP1 (SLC11A1) and TLR4, known to be involved in the level of chicken infection 3 d after inoculation by Salmonella were thus tested on a total of 331 hens orally inoculated at the peak of lay with 10(9) bacteria. The animals and their parents were genotyped for a total of 10 microsatellite markers mapped on chromosomes 7 and 17. Using maximum likelihood analysis and interval mapping, it was found that the SLC11A1 region was significantly involved in the control of the probability of spleen contamination 4 wk after inoculation. Single nucleotide polymorphisms (SNP) within the SLC11A1 and TLR4 gene were tested on those animals as well as on a second batch of 279 hens whose resistance was assessed in the same conditions. As the former was significantly associated with the risk of spleen contamination and the number of contaminated organs, SLC11A1 appears to be involved in the control of resistance to Salmonella carrier state. The involvement of the TLR4 gene was also highly suspected as a significant association between SNP within the gene, and the number of contaminated organs was detected.

Genetic resistance to mortality of day-old chicks and carrier-state of hens after inoculation with Salmonella enteritidis.

The heritability of resistance of poultry to Salmonella enteritidis (SE) was investigated. Three measurements of resistance were made: survival after intramuscular inoculation of 419 day-old chicks, absence versus presence of Salmonella 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 mutant of this serotype. In the first two models of infection, resistance appeared to be heritable. The heritability was estimated from the sire and dam components, respectively, at 0.14 ± 0.10 and 0.62 ± 0.16 for chick mortality, 0.47 ± 0.21 and 0.13 ± 0.26 for resistance to spleen contamination, and 0.24 ± 0.15 and 0.53 ± 0.26 for resistance to caecal contamination in laying hens. By contrast the estimated heritability of antibody response was very low (0.03 ± 0.08 and 0.10 ± 0.08 when estimated from the sire and dam components, respectively). These results suggest that a selection for increased resistance to SE may be efficient.

Variability in the resistance of four chicken lines to experimental intravenous infection with Salmonella enteritidis phage type 4.

The capacity of four chicken lines (Y11, L2, B13, PA12) to control Salmonella enteritidis (SE) phage type 4 (PT4) systemic colonization was investigated. Thirteen-week-old chickens were intravenously inoculated with 10(6) SE colony-forming units, and the levels of SE colonization were determined at various time intervals after inoculation in liver, spleen, genital organs, and ceca. The course of SE infection showed a rapid contamination of liver, spleen, and genital organs, whereas the ceca were infected later. A significant (P < 0.001) effect of the chicken line on levels of SE was detected on day 3 postinoculation (PI) in liver and ceca, on day 10 PI in ceca, and on day 15 PI in spleen. Because an early control of systemic Salmonella infection by the Ity/Nramp1 gene has been demonstrated in mice, we aimed to study the early resistance of chickens to SE. As a consequence, we then focused our study on the between- and within-line variabilities of SE levels on day 3 PI. According to the SE levels in liver on day 3 PI, the chicken lines could be classified as susceptible (Y11 and L2) or resistant (PA12 and B13). This early variability was explored in resistant B13 and susceptible L2 lines. Differences between these two lines were confirmed in liver but not in ceca. A large within-line variability was observed in all organs of these two lines. The genetic origin of this variability will have to be determined as a prerequisite to an eventual selection.

Line differences in resistance to Salmonella enteritidis PT4 infection.

1. Four groups of hens, each of a different line, were inoculated at peak of lay, per os in the crop with 1 ml of a suspension containing 10(9) cfu/ml Salmonella enteritidis PT4 (SE). The kinetics of SE contamination in the environment, egg shell and yolk were studied during the first 28 d post inoculation. On the day of slaughter, intestines, caeca, spleen, liver, ovary, oviduct and content were investigated for SE contamination. 2. The commercial egg-type line L2 was found to be the most susceptible to SE. It laid many SE-positive yolks (13.8%) and internal and faecal organs were frequently infected. 3. Certain lines are found to exhibit a degree of resistance to SE; the cause of which is unknown and might be attributed to major genes.

Comparison of resistance of various poultry lines to infection by Salmonella enteritidis.

A study was undertaken to determine the susceptibility or resistance of 9 outbred experimental or commercial poultry lines to Salmonella enteritidis PT4. Young chicks were inoculated either intramuscularly or orally just after hatching. After intramuscular challenge the lines could be divided into susceptible lines (LD 50% < or = 10(2) Salmonella per animal), intermediate lines (LD 50% about 10(4) Salmonella) and resistant lines (LD 50% > 10(5) Salmonella). The results obtained after oral challenge confirmed these 3 groups for both mortality rates and the probability of the presence of salmonellae in the spleen and liver. There was no difference between lines concerning caecal carriage.

Isolation, characterisation and preliminary cross-protection studies with a new pathogenic avian infectious bronchitis virus (strain PL-84084).

Virological 1 examination of a severe infectious bronchitis (IB)-like field case in laying hens, led to the isolation of a coronavirus antigenically different from Massachusetts, Connecticut and four Dutch IB variant strains. The virulence of the isolate for the fowl, and its dual tropism for the respiratory and genital tracts were demonstrated. In preliminary cross-protection studies Commercial vaccines did not protect against challenge with this isolate. These points and the possible economic significance of the virus are discussed.