Broiler lines divergently selected for digestive efficiency also differ in their susceptibility to colibacillosis.

Increasing feed efficiency of broiler chickens by selective breeding could lead to decreased feed cost and reduced environmental impact of poultry production. At INRA, two broiler chicken lines (D+/D-) were divergently selected for their digestive efficiency. Strong differences were shown between both lines for the anatomy and histology of the digestive tract, and for the intestinal microbiota composition. In the present study, we investigated whether this selection also had an effect on susceptibility to colibacillosis, which is one of the main causes of economic losses in poultry production. The broiler lines D+/D- were challenged with an avian pathogenic Escherichia coli strain. A first experiment was conducted to assess the 50% lethal dose by subcutaneous infection of hatchlings, whereas a second experiment reproduced colibacillosis by infecting air sacs of 23-day-old chicks. The 50% lethal dose was very low for both lines. However, the line with the higher digestive efficiency (D+) was the less susceptible to colibacillosis. This result is interesting for selection purposes and opens the way to integrative genetic studies of the interactions between digestion efficiency and resistance to colibacillosis.

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.

Host cytokine response and resistance to Salmonella infection.

Knowledge of the host response, of the resistance process, and of the mediators committed against Salmonella infection is essential to progress towards better means of prophylaxis and eradication. In this context, the present contribution attempts to interconnect, with the pivotal role of the macrophage, the early resistance process under the control of the Nramp1 gene and the cytokine response for resolving infection. IL-12 produced by macrophages is an inducer of IFN-gamma production, which in turn activates the macrophage antibacterial activity and synergizes its effects with TNF-alpha. All three of these cytokines are powerful actors in the first line of anti-Salmonella defence. It can be pointed out that susceptible and resistant individuals do not seem to see the cytokine environment the same way, the former being unresponsive to IL-1 or GM-CSF treatment and deficient in IFN-gamma production. These discrepancies may rely on cell signalling events that could be defective in macrophages of the susceptible phenotype.

Effect of the mouse Nramp1 genotype on the expression of IFN-gamma gene in early response to Salmonella infection.

The early interferon-gamma (IFN-gamma) response after Salmonella infection was compared in resistant and susceptible congenic mice strains differing by the allele of Nramp1. IFN-gamma gene expression in vivo was studied by quantitative reverse transcription-PCR performed from the spleen of mice challenged by Salmonella either in multiplication or in resting phase. The spleen colonisation by Salmonella in vivo was investigated and was shown to be identical in a genetically defined host, whatever the initial phase of the bacteria. However, the bacterial load of the spleen was significantly reduced in Nramp1(r) (resistant) mice compared to Nramp1(s) (susceptible) mice. The background level of IFN-gamma mRNA was higher in the spleen of resistant mice than in the susceptible mice, before infection. Interestingly, the early upregulation of IFN-gamma gene transcription, which was observed after infection with Salmonella, was reproducibly delayed in susceptible mice compared to resistant mice. Finally, the kinetics of the host IFN-gamma response seems to be Nramp1 dependent. Resistant mice present the advantage of being more prompt to express this anti-infectious cytokine gene than susceptible mice in response to Salmonella infection.

Quantification of ovine cytokine gene expression by a competitive RT-PCR method.

A quantitative competitive RT-PCR method was developed in order to measure IL-1beta, IL-4, IL-12, IFNgamma, TNFalpha and G(3)PDH mRNA from samples of ovine tissue such as lymph node or spleen. The main advantage of the method relies on the use, for each target sequence, of an internal competitor construct similar to the relevant target, but 4-bp different in size. This competitive strategy is validated by the equivalence of the amplification process, observed separately between competitor DNA and target DNA species. Furthermore, the copy number of each cytokine cDNA is normalized to a fixed copy number of G(3)PDH cDNA. The cDNA level of this constitutive gene was effectively shown to remain constant whatever the tissue studied and independently of the experimental conditions used. The accurate and reproducible data obtained permit the application of this quantitative RT-PCR method to measure the sheep cytokine response to Salmonella infection. Early induction of IFNgamma mRNA was observed in the draining lymph node 1 day after infection. At the same time, a strong increase of IL-1beta mRNA was observed in local and systemic lymphoid organs, suggesting the initiation of the inflammatory response. Finally, the overall results demonstrate the efficiency of the method and its suitability for further studies of the immune response in the ovine species.

The interleukin-1 system and female reproduction.

Interleukins (ILs) are known best for their involvement in the immune system and their role during inflammation. In the ovary, a growing body of evidence suggests that the ovarian follicle is a site of inflammatory reactions. Thus ovarian cells could represent sources and targets of ILs. Since then, the IL-1 system components (IL-1alpha, IL-1beta, IL-1 receptor antagonist, IL-1 receptors) have been demonstrated to have several sites of synthesis in the ovary. These factors have been localized in the various ovarian cell types, such as the oocyte, granulosa and theca cells, in several mammalian species. IL-1-like bioactivity has been reported in human and porcine follicular fluid at the time of ovulation. The role of IL-1 in local processes is still poorly known, although there is evidence for involvement in the ovulation process, and in oocyte maturation. More precisely, IL-1 may be involved in several ovulation-associated events such as the synthesis of proteases, regulation of plasminogen activator activity, prostaglandin and nitric oxide production. IL-1 also regulates ovarian steroidogenesis. These different aspects of the involvement of the IL-1 system in important aspects of female reproduction are discussed.

Cytokine gene expression in lymph node and spleen of sheep in response to Salmonella infection by two serotypes displaying different host specificity.

In order to investigate the determinism of the host specificity and to better understand the host resistance mechanisms, infections of sheep were performed with either S. abortusovis, serotype specific for ovine species, or with S. dublin, serotype adapted to cattle and accidentally transmissible to human. Following a subcutaneous challenge, S. dublin disseminated more rapidly towards lymphoid tissues than S. abortusovis. However, S. abortusovis tended to persist in spleen more efficiently than S. dublin. Using a quantitative RT-PCR method, the expression level of ovine cytokines genes was measured in the draining lymph node and in the spleen, in the course of infection. Inflammatory cytokine response was characterised by an early and strong increase of IL-1beta and TNFalpha mRNA in both lymphoid organs following S. dublin infection, while S. abortusovis challenge only induced IL-1beta mRNA increase in the spleen at day 3 post-inoculation. Likewise, S. dublin infection provoked a marked increase of IL-12 mRNA and a slight up-regulation of IFNgamma gene transcription in the local lymphoid site, in contrast to S. abortusovis infection. Elsewhere, both serotypes induced a strong and early IL-10 mRNA production and had no effect on IL-4 gene expression. Finally, taken together, these data suggest that the intensity of inflammatory and anti-infectious cytokine responses, but not the type 2 cytokine response, is serotype-dependent. They also suggest that the host-specific serotype, by limiting the host cytokine-mediated defence, could favour its persistence within lymphoid organs.

Emergency and therapeutic vaccination--is stimulating innate immunity an option?

There is increasing evidence that activation of innate immunity, in animals and man, by live vaccines, sub-unit vaccines or synthetic or non-synthetic stimulants can induce a profound and rapidly induced resistance to pathogens, including infectious agents that are unrelated to the stimulating antigen or agent. We review the evidence for this phenomenon and present the proposition that this approach might be used to stimulate immunity during the life of the animal when susceptibility to infection is high and when normal vaccination procedures may be inappropriate.