Gallinaceous Poultry Research Articles

Ducks: The “Trojan Horses” of H5N1 influenza

Wild ducks are the main reservoir of influenza A viruses that can be transmitted to domestic poultry and mammals, including humans. Of the 16 hemagglutinin (HA) subtypes of influenza A viruses, only the H5 and H7 subtypes cause highly pathogenic (HP) influenza in the natural hosts. Several duck species are naturally resistant to HP Asian H5N1 influenza viruses. These duck species can shed and spread virus from both the respiratory and intestinal tracts while showing few or no disease signs. While the HP Asian H5N1 viruses are 100% lethal for chickens and other gallinaceous poultry, the absence of disease signs in some duck species has led to the concept that ducks are the "Trojan horses" of H5N1 in their surreptitious spread of virus. An important unresolved issue is whether the HP H5N1 viruses are maintained in the wild duck population of the world. Here, we review the ecology and pathobiology of ducks infected with influenza A viruses and ducks' role in the maintenance and spread of HP H5N1 viruses. We also identify the key questions about the role of ducks that must be resolved in order to understand the emergence and control of pandemic influenza. It is generally accepted that wild duck species can spread HP H5N1 viruses, but there is insufficient evidence to show that ducks maintain these viruses and transfer them from one generation to the next.

Pathogenesis and pathobiology of avian influenza virus infection in birds

Avian influenza (AI) viruses vary in their ability to produce infection, disease and death in different bird species. Based on the pathobiological effect in chickens, AI viruses (AIV) are categorised as low pathogenic (LPAIV) or highly pathogenic (HPAIV). Typically, LPAIV cause asymptomatic infections in wild aquatic birds, but when introduced into domesticated poultry, infections may be asymptomatic or produce clinical signs and lesions reflecting pathophysiological damage to the respiratory, digestive and reproductive systems. The HPAIV have primarily been seen in gallinaceous poultry, producing high morbidity and mortality, and systemic disease with necrosis and inflammation in multiple visceral organs, nervous and cardiovascular systems, and the integument. Although HPAIV have rarely infected domestic waterfowl or wild birds, the Eurasian-African H5N1 HPAIV have evolved over the past decade with the unique capacity to infect and cause disease in domestic ducks and wild birds, producing a range of syndromes including asymptomatic respiratory and digestive tract infections; systemic disease limited to two or three critical organs, usually the brain, heart and pancreas; and severe disseminated infection and death as seen in gallinaceous poultry. Although experimental studies using intranasal inoculation have produced infection in a variety of wild bird species, the inefficiency of contact transmission in some of them, for example, passerines and Columbiformes, suggests they are unlikely to be a reservoir for the viruses, while others such as some wild Anseriformes, can be severely affected and could serve as a dissemination host over intermediate distances.

Efficacy of a commercial inactivated H5 influenza vaccine against highly pathogenic avian influenza H5N1 in waterfowl evaluated under field conditions

Highly pathogenic avian influenza virus (HPAIV) can cause devastating losses in the poultry industry. In addition, several HPAIV exhibit a zooanthroponotic potential and can cause fatal infections in humans. These attributes particularly apply to HPAIV H5N1 of Asian origin. Due to the absence of overt clinical symptoms, introduction and subsequent spread of HPAIV H5N1 in domestic waterfowl (especially ducks) may occur undetected, which increases the risk of transspecies transmissions to highly vulnerable gallinaceous poultry and mammals, including humans. Humans may also become infected with HPAIV H5N1 by food products from slaughtered, silently infected ducks. Vaccination against HPAIV can raise a protective barrier against an incursion of HPAIV since, at least under experimental conditions, the reproduction factor R0 lowered to <1, which ensures eradication of the virus. The objective of this study was to analyse whether these results can also be obtained under free-ranging field conditions in commercially reared flocks of goose parents and fattening ducks injected with a licensed, adjuvanted inactivated H5N2 vaccine. The time and labour required for the vaccination of these geese and duck flocks exceeded expected values, mainly due to animal sorting according to foot ring labels. No adverse effects directly associated with vaccination were observed. Serologically, a homogenous H5-specific antibody response was induced. Titres varied with temporal distance from the last application of vaccine. Geese parents were clinically protected against challenge with HPAIV A/Cygnus cygnus/Germany/R65/06 (H5N1), but still could be infected and spread HPAIV H5N1, albeit at lower levels and for shorter periods compared to unvaccinated controls. Fattening Pekin ducks proved to be clinically resistant against challenge virus infection and shed very little virus.

Modulation of the severity of highly pathogenic H5N1 influenza in chickens previously inoculated with Israeli H9N2 influenza viruses

The continued evolution of H9N2 and H5N1 viruses and their spread and re-emergence across Eurasia raise concern that prior H9N2 virus infection may limit the detection of subsequent H5N1 infection in gallinaceous poultry by attenuating the severity of disease. We show that H9N2 viruses isolated from Israeli turkeys during 2000–2004 were antigenically and genetically distinguishable. These three H9N2 viruses caused no overt signs of disease in chickens. The 2004 isolate replicated and spread most efficiently, and chickens previously inoculated with this H9N2 virus showed 90%–100% survival after inoculation 1 to 35 days later with lethal H5N1 virus. Chickens that survived did not show signs of disease but did shed lethal H5N1 virus from the cloaca. The modulation of survivability was time-dependent; the effect was maximal 5 days after H9N2 inoculation. These findings suggest that co-circulation of H9N2 viruses can contribute to the spread of lethal H5N1 viruses.

Using Mean Infectious Dose of High- and Low-Pathogenicity Avian Influenza Viruses Originating from Wild Duck and Poultry as One Measure of Infectivity and Adaptation to Poultry

The mean infectious doses of selected avian influenza virus (AIV) isolates, determined in domestic poultry under experimental conditions, were shown to be both host-dependent and virus strain-dependent and could be considered one measure of the infectivity and adaptation to a specific host. As such, the mean infectious dose could serve as a quantitative predictor for which strains of AIV, given the right conditions, would be more likely transmitted to and maintained in a given species or subsequently cause an AI outbreak in the given species. The intranasal (IN) mean bird infectious doses (BID50) were determined for 11 high-pathogenicity AIV (HPAIV) isolates of turkey and chicken origin for white leghorn (WL) chickens, and for low-pathogenicity AIV (LPAIV) isolates of chicken (n = 1) and wild mallards (n = 2) for turkeys, and WL and white Plymouth rock (WPR) chickens, domestic ducks and geese, and Japanese quail. The BID50 for HPAIV isolates for WL chickens ranged from 10(1.2) to 10(4.7) mean embryo infectious dose (EID50) (median = 10(2.9)). For chicken-origin HPAIV isolates, the BID50 in WL chickens ranged from 10(1.2) to 10(3.0) EID50 (median = 10(2.6)), whereas for HPAIV isolates of turkey origin, the BID50 in WL chickens was higher, ranging from 10(2.8) to 10(4.7) EID50 (median = 10(3.9)). The BID50 of 10(4.7) was for a turkey-origin HPAIV virus that was not transmitted to chickens on the same farm, suggesting that, under the specific conditions present on that farm, there was insufficient infectivity, adaptation, or exposure to that virus population for sustained chicken transmission. Although the upper BID50 limit for predicting infectivity and sustainable transmissibility for a specific species is unknown, a BID50 < 10(4.7) was suggestive of such transmissibility. For the LPAIVs, there was a trend for domestic ducks and geese and Japanese quail to have the greatest susceptible and for WL chickens to be the most resistant, but turkeys were susceptible to two LPAIV tested when used at moderate challenge doses. This suggests domestic ducks and geese, turkeys, and Japanese quail could serve as bridging species for LPAIVs from wild waterfowl to chickens and other gallinaceous poultry. These data do provide support for the commonly held and intuitive belief that mixing of poultry species during rearing and in outdoor production systems is a major risk factor for interspecies transmission of AIVs and for the emergence of new AIV strains capable of causing AI outbreaks because these situations present a more diverse host population to circumvent the natural host dependency or host range of circulating viruses.

Evidence of Infection by H5N2 Highly Pathogenic Avian Influenza Viruses in Healthy Wild Waterfowl

The potential existence of a wild bird reservoir for highly pathogenic avian influenza (HPAI) has been recently questioned by the spread and the persisting circulation of H5N1 HPAI viruses, responsible for concurrent outbreaks in migratory and domestic birds over Asia, Europe, and Africa. During a large-scale surveillance programme over Eastern Europe, the Middle East, and Africa, we detected avian influenza viruses of H5N2 subtype with a highly pathogenic (HP) viral genotype in healthy birds of two wild waterfowl species sampled in Nigeria. We monitored the survival and regional movements of one of the infected birds through satellite telemetry, providing a rare evidence of a non-lethal natural infection by an HP viral genotype in wild birds. Phylogenetic analysis of the H5N2 viruses revealed close genetic relationships with H5 viruses of low pathogenicity circulating in Eurasian wild and domestic ducks. In addition, genetic analysis did not reveal known gallinaceous poultry adaptive mutations, suggesting that the emergence of HP strains could have taken place in either wild or domestic ducks or in non-gallinaceous species. The presence of coexisting but genetically distinguishable avian influenza viruses with an HP viral genotype in two cohabiting species of wild waterfowl, with evidence of non-lethal infection at least in one species and without evidence of prior extensive circulation of the virus in domestic poultry, suggest that some strains with a potential high pathogenicity for poultry could be maintained in a community of wild waterfowl.

Understanding the Complex Pathobiology of High Pathogenicity Avian Influenza Viruses in Birds

Avian influenza (AI) viruses are a diverse group of viruses that can be divided into 144 subtypes, based on different combinations of the 16 hemagglutinin and nine neuraminidase subtypes, and two pathotypes (low and high pathogenicity [HP]), based on lethality for the major poultry species, the chicken. However, other criteria are important in understanding the complex biology of AI viruses, including host adaptation, transmissibility, infectivity, tissue tropism, and lesion, and disease production. Overall, such pathobiological features vary with host species and virus strain. Experimentally, HPAI viruses typically produce a similar severe, systemic disease with high mortality in chickens and other gallinaceous birds. However, these same viruses usually produce no clinical signs of infection or only mild disease in domestic ducks and wild birds. Over the past decade, the emergent HPAI viruses have shifted to increased virulence for chickens as evident by shorter mean death times and a greater propensity for massive disseminated replication in vascular endothelial cells. Importantly, the Asian H5N1 HPAI viruses have changed from producing inconsistent respiratory infections in 2-wk-old domestic ducks to some strains being highly lethal in ducks with virus in multiple internal organs and brain. However, the high lethality for ducks is inversely related to age, unlike these viruses in gallinaceous poultry, which are highly lethal irrespective of the host age. The most recent Asian H5N1 HPAI viruses have infected some wild birds, producing systemic infections and death. Across all bird species, the ability to produce severe disease and death is associated with high virus replication titers in the host, especially in specific tissues such as brain and heart.

Using meteorological data to determine the risk of heat stress

This paper first proposes a technique of computing air temperature and humidity in stables based on outdoor air parameters and biological production of animals. The computation technique is outlined. The calculated values are then used to assess the potential of evaporation cooling in mild climatic conditions. Graphs illustrate the assumed effect of evaporation cooling equipment inside a stable housing of egg laying hens. Used in the computation were hourly meteorological readings obtained during the period May to August in years 2000 to 2002, in the locality with a potential installation of a cooling system. Other Graphs illustrate the time the animals spent in an environment with a particular air temperature. For instance in June 2002, the time animals in the stable were exposed to temperatures 27&amp;deg;C or higher was reduced by using an air cooling system from 39 h to 22 h, and in July 2002 from 33 h to 4 h. The envisaged model can be modified for other kinds of gallinaceous poultry and pigs.

Phylogenetic analyses of type A influenza genes in natural reservoir species in North America reveals genetic variation

The matrix (M) (98 isolates), nucleoprotein (NP) (67 isolates), non-structural (NS) (107 isolates), H4 subtype HA1 (21 isolates) and H6 (38 isolates) subtype HA1 region of the hemagglutinin (HA) gene were sequenced from avian influenza viruses isolated from North American wild aquatic birds between 1969 and 2003. Sequences were compared phylogenetically with all available wild aquatic bird isolate sequences and sequences from isolates from gallinaceous poultry and mammalian host species. Among the wild aquatic bird isolates the NS subtype B NS1 and subtype B NS2 proteins were the most conserved with minimum amino acid identities of 98.0 and 99.0%, respectively. The H6 HA1, M and NS subtype A genes were more divergent; both nucleotide and amino acid divergence levels were similar to those of the same genes from gallinaceous poultry and mammalian origin type A influenza isolates. Phylogenetically none of the genes assorted geographically (within North America), chronologically or by species of origin, unlike isolates from non-natural host species, suggesting that selection pressure is low. A lack of clear temporal or spatial grouping shows that multiple lineages of virus co-circulate and precludes the development of an epidemiological map for influenza virus from these species.

Investigation of outbreaks of highly pathogenic H5N1 avian influenza in waterfowl and wild birds in Hong Kong in late 2002

Outbreaks of highly pathogenic H5N1 avian influenza have occurred in Hong Kong in chickens and other gallinaceous poultry in 1997, 2001, twice in 2002 and 2003. High mortality rates were seen in gallinaceous birds but not in domestic or wild waterfowl or other wild birds until late 2002 when highly pathogenic H5N1 avian influenza occurred in waterfowl (geese, ducks and swans), captive Greater Flamingo (Phoenicopterus ruber) and other wild birds (Little Egret Egretta garzetta) at two waterfowl parks and from two dead wild Grey Heron (Ardea cinerea) and a Black-headed Gull (Larus ridibundus) in Hong Kong. H5N1 avian influenza virus was also isolated from a dead feral pigeon (Columba livia) and a dead tree sparrow (Passer montanus) during the second outbreak. The first waterfowl outbreak was controlled by immediate strict quarantine and depopulation 1 week before the second outbreak commenced. Control measures implemented for the second outbreak included strict isolation, culling, increased sanitation and vaccination. Outbreaks in gallinaceous birds occurred in some live poultry markets concurrently with the second waterfowl outbreak, and infection on a chicken farm was detected 1 week after the second waterfowl park outbreak was detected, on the same day the second grey heron case was detected. Subsequent virus surveillance showed the outbreaks had been contained.