Comparative evaluation of immune responses of swine in PRRS-stable and unstable herds.

Porcine Reproductive and Respiratory Syndrome (PRRS) is an elusive model of host/virus relationship in which disease is determined by virus pathogenicity, pig breed susceptibility and phenotype, microbial infectious pressure and environmental conditions. Successful disease control in PRRS-endemic Countries corresponds to "stability", i.e. a condition with no clinical signs of PRRS in the breeding-herd population and no viremia in weaning-age pigs. The aim of this work was to compare the profile and time-course of humoral and cell-mediated immunity in stable and unstable herds, respectively. In particular, we investigated PRRS virus (PRRSV) in serum and group oral fluid samples by Real-time RT-PCR, PRRSV-specific IgA and IgG in oral fluids, serum IgG antibody and the cell-mediated response (PRRSV-specific release of interferon-gamma) in whole blood samples. These parameters were measured in order to identify possible discrepancies in the development and kinetics of the immune response against PRRSV. PRRS-free gilts got regularly infected after entering PRRS-stable and unstable farms. In an open cycle, unstable pig farm PRRSV infection could be demonstrated in all groups of pigs, including suckling piglets. Four main results should be highlighted: A) the precocity of the Ab response in group oral fluids was generally similar to that recorded in sera; B) circulation of PRRSV was consistently detected in all age groups in the unstable herds, as opposed to the stable ones; C) an early, balanced, IgA and IgG response in oral fluids was only observed in the stable herds; D) an early IFN-gamma response after PRRSV infection was often observed in stable herds, as opposed to the unstable ones. These were characterized by IFN-gamma responses in piglets, likely due to transfer of maternal immunity. Most important, the mucosal IgA response was associated with cessation of virus excretion in oral fluid samples of PRRS-unstable herds. The above findings indicate that a peculiar profile of immune response to PRRSV can be found in PRRS-stable herds. Therefore, the outlined immune parameters can represent a useful readout system to evaluate successful adaptation to PRRSV based on acclimatization of breeding animals and management of pig flow.

Salmonella serovar-specific interaction with jejunal epithelial cells.

Gut is often a receptacle for many different pathogens in feed and/or the environment, such as Salmonella spp. The current knowledge about pathogenicity of Salmonella is restricted to few serotypes, whereas other important ones like S. Coeln, S. Thompson, S. Veneziana, have not been investigated yet in human and animal models. Therefore, the aim of our work was to verify the ability of widespread environmental Salmonella strains to penetrate and modulate innate immunity in pig intestinal IPEC-J2 cells. Our results outline the different ability of Salmonella strains to modulate innate immunity; the expression of the IFN-ß gene was increased by S. Typhimurium, S. Ablogame and S. Diarizonae 2, that also caused an inflammatory response in terms of Interleukin (IL)-1ß and/or IL-8 gene espression. In particular, IL-8 gene expression and protein release were significantly modulated by 5 Salmonella strains out of 7. Interestingly, S. Typhimurium, S. Coeln and S. Thompson strains, characterized by a peculiar ability to penetrate into IPEC-J2 cells, up-regulated both IL-8 and TNF-α gene expression. Accordingly, blocking IL-8 was shown to decrease the penetration of S. Typhimurium. On the contrary, S. Diarizonae strain 1, showing lesser invasion of IPEC-J2 cells, down-regulated the p38-MAPK pathway, and it did not induce an inflammatory response. Our results confirm that IPEC-J2 cells are a useful model to evaluate host-gut pathogen interaction and indicate IL-8 and TNF-α as possible predictive markers of invasiveness of Salmonella strains in enterocytes.

Expression Dynamics of Innate Immunity in Influenza Virus-Infected Swine.

The current circulating swine influenza virus (IV) subtypes in Europe (H1N1, H1N2, and H3N2) are associated with clinical outbreaks of disease. However, we showed that pigs could be susceptible to other IV strains that are able to cross the species barrier. In this work, we extended our investigations into whether different IV strains able to cross the species barrier might give rise to different innate immune responses that could be associated with pathological lesions. For this purpose, we used the same samples collected in a previous study of ours, in which healthy pigs had been infected with a H3N2 Swine IV and four different H3N8 IV strains circulating in different animal species. Pigs had been clinically inspected and four subjects/group were sacrificed at 3, 6, and 21 days post infection. In the present study, all groups but mock exhibited antibody responses to IV nucleoprotein protein. Pulmonary lesions and high-titered viral replication were observed in pigs infected with the swine-adapted virus. Interestingly, pigs infected with avian and seal H3N8 strains also showed moderate lesions and viral replication, whereas equine and canine IVs did not cause overt pathological signs, and replication was barely detectable. Swine IV infection induced interferon (IFN)-alpha and interleukin-6 responses in bronchoalveolar fluids (BALF) at day 3 post infection, as opposed to the other non-swine-adapted virus strains. However, IFN-alpha responses to the swine-adapted virus were not associated with an increase of the local, constitutive expression of IFN-alpha genes. Remarkably, the Equine strain gave rise to a Serum Amyloid A response in BALF despite little if any replication. Each virus strain could be associated with expression of cytokine genes and/or proteins after infection. These responses were observed well beyond the period of virus replication, suggesting a prolonged homeostatic imbalance of the innate immune system.

Cross-Species Infectivity of H3N8 Influenza Virus in an Experimental Infection in Swine.

UNLABELLED: Avian influenza A viruses have gained increasing attention due to their ability to cross the species barrier and cause severe disease in humans and other mammal species as pigs. H3 and particularly H3N8 viruses, are highly adaptive since they are found in multiple avian and mammal hosts. H3N8 viruses have not been isolated yet from humans; however, a recent report showed that equine influenza A viruses (IAVs) can be isolated from pigs, although an established infection has not been observed thus far in this host. To gain insight into the possibility of H3N8 avian IAVs to cross the species barrier into pigs, in vitro experiments and an experimental infection in pigs with four H3N8 viruses from different origins (equine, canine, avian, and seal) were performed. As a positive control, an H3N2 swine influenza virus A was used. Although equine and canine viruses hardly replicated in the respiratory systems of pigs, avian and seal viruses replicated substantially and caused detectable lesions in inoculated pigs without previous adaptation. Interestingly, antibodies against hemagglutinin could not be detected after infection by hemagglutination inhibition (HAI) test with avian and seal viruses. This phenomenon was observed not only in pigs but also in mice immunized with the same virus strains. Our data indicated that H3N8 IAVs from wild aquatic birds have the potential to cross the species barrier and establish successful infections in pigs that might spread unnoticed using the HAI test as diagnostic tool. IMPORTANCE: Although natural infection of humans with an avian H3N8 influenza A virus has not yet been reported, this influenza A virus subtype has already crossed the species barrier. Therefore, we have examined the potential of H3N8 from canine, equine, avian, and seal origin to productively infect pigs. Our results demonstrated that avian and seal viruses replicated substantially and caused detectable lesions in inoculated pigs without previous adaptation. Surprisingly, we could not detect specific antibodies against hemagglutinin in any H3N8-infected pigs. Therefore, special attention should be focused toward viruses of the H3N8 subtype since they could behave as stealth viruses in pigs.

Treatment of eggshell abnormalities and reduced egg production caused by Mycoplasma synoviae infection.

Eggshell abnormalities were seen in the apex of eggs in two of three flocks of multi-age, Hy-Line layer chickens housed on a farm in Northern Italy. Approximately 1.3% to 1.8% of eggs in one flock were affected, amounting to 300-400 eggs per day; the abnormalities resulted in a great deal of breakage and spoilage of healthy eggs. The mean weight of eggs was also reduced. Egg abnormalities in a second flock were less severe. Mycoplasma synoviae was detected in birds from both of the affected flocks by serologic, cultural, and molecular techniques, but not in a third, adjacent flock where no eggshell abnormalities were seen. Treatment with tylosin, administered in the drinking water for 5 days, resulted in an immediate improvement of eggshell quality and egg weight. There was no evidence of infectious bronchitis virus in the flocks.