Detection of influenza A viruses in eggs laid by infected turkeys.

Studies are limited on evaluating the potential of influenza viruses for egg-borne dissemination. In our previous studies, experimental infection of breeder turkeys with A/turkey/Ohio/313053/04 resulted in drastic declines in egg production, and we confirmed high levels of virus replication and an abundant distribution of avian-specific alpha2,3 sialic acid-gal receptors in the oviduct of these turkeys. In the present study, following experimental inoculation of A/turkey/Ohio/313053/04 in breeder turkeys, we detected these viruses in the albumin of eggs using real-time RT-PCR (RRT-PCR) and virus isolation in embryonated chicken eggs. Swabs from egg shells were also found positive by RRT-PCR. This is the first report of the detection of low pathogenic influenza viruses from internal egg contents following experimental infection. The possibility of hatchery contamination by egg-borne influenza viruses, and the spread of virus during movement of contaminated cracked eggs and egg flats, pose concerns regarding viral dissemination of influenza.

The high susceptibility of turkeys to influenza viruses of different origins implies their importance as potential intermediate hosts.

Several previous reports and our studies show that waterfowl-origin influenza viruses can be more easily transmitted to domestic turkeys than chickens. Similarly, studies indicate turkeys to be better hosts for low pathogenic avian influenza viruses isolated from commercial poultry operations and live bird markets in comparison to chickens. Low 50% infectious-dose titers of wild bird as well as poultry-adapted viruses for turkeys further suggest that turkeys can be easily infected following a low-dose exposure. Also, interspecies transmission of swine influenza viruses to turkeys occurs frequently. These findings suggest the role of turkeys as suitable intermediate hosts that can be easily infected with influenza viruses of different origins and that turkeys can act as source of infection for other land-based poultry or even mammals.

Pathobiological characterization of low-pathogenicity H5 avian influenza viruses of diverse origins in chickens, ducks and turkeys.

We undertook one of the most comprehensive studies on the replication and intraspecies transmission characteristics of low-pathogenicity avian influenza viruses in ducks, chickens and turkeys. Our results indicated that most of these isolates could replicate and be transmitted in poultry without inducing clinical disease. However, differences in transmission to contact control birds were noted, emphasizing the importance of having contact control cage mates in biological characterization experiments. Ducks supported the replication of viruses of wild aquatic bird origin in their respiratory and digestive tracts equally well. The viruses from wild aquatic birds were not effectively transmitted among chickens. In contrast, the wild-bird isolates and viruses of domestic bird origin from live-bird markets and commercial poultry operations replicated and were transmitted more efficiently in turkeys than in chickens or ducks. We also found a lower minimal infectious dose requirement for infection of turkeys compared to chickens and ducks. Our data support an important role of turkeys as being more susceptible hosts for avian influenza viruses than domestic ducks and chickens. These results highlight the role of turkeys as intermediate or bridging hosts in the transmission of influenza viruses from wild birds to land-based domestic poultry or among different land-based bird species.

Pathobiology of triple reassortant H3N2 influenza viruses in breeder turkeys and its potential implication for vaccine studies in turkeys.

Triple reassortant (TR) H3N2 influenza viruses have been isolated from turkeys in the United States since 2003. These TR H3N2 virus infections have been associated with drastic declines in egg production in breeder turkeys although co-infection with multiple agents could have been responsible for exacerbating the clinical signs. In this study, we experimentally confirmed that TR H3N2 influenza virus alone can cause drastic reduction/complete cessation of egg production and pathology of the reproductive tract in 26-week-old breeder turkeys. We confirmed high levels of virus replication and abundant distribution of avian specific alpha2,3 sialic acid-galactose receptors in the oviduct of these turkeys. Although 2-6-week-old turkeys are routinely used for pathogenicity and vaccine protection studies, the low levels of viral shedding and asymptomatic infections in this age group often pose difficulty in interpretation of results. Our study shows that breeder turkeys should be used to assess the potential pathogenicity of TR H3N2 viruses and the viral titers and pathology of the oviduct as well as egg production data can be good measures of protection following in vivo challenge in vaccine efficacy studies.

Pathogenicity and transmission studies of H5N2 parrot avian influenza virus of Mexican lineage in different poultry species.

In 2004, a low pathogenic H5N2 influenza virus (A/parrot/CA/6032/04) was identified in a psittacine bird for the first time in the United States. Sequence and phylogenetic analysis of the hemagglutinin gene grouped the parrot isolate under the Mexican lineage H5N2 viruses (subgroup B) with highest similarity to recent chicken-origin isolates from Guatemala. Antigenic analysis further confirmed the close relatedness of the parrot isolate to Mexican lineage viruses, the highest cross-reactivity being demonstrated to Guatemala isolates. In vivo studies of the parrot isolate in chickens, ducks and turkeys showed that the virus, though did not cause any clinical signs, could replicate to high titers in these birds and efficiently transmit to contact control cage mates. The possibility that the parrot harboring the virus was introduced into the United States as a result of illegal trade across the border provides additional concern for the movement of foreign animal diseases from neighboring countries. Considering the potential threat of the virus to domestic poultry, efforts should be continued to prevent the entry and spread of influenza viruses by imposing effective surveillance and monitoring measures.