Bagaza virus is pathogenic and transmitted by direct contact in experimentally infected partridges,but is not infectious in house sparrows and adult mice

Bagaza virus (BAGV) is a mosquito-borne flavivirus belonging to the Ntaya serocomplex. In 2010, a disease outbreak was reported in Cádiz (Southern Spain) affecting game birds (red-legged partridges and common pheasants). In this work, red-legged partridges were inoculated experimentally with infectious BAGV isolated from this outbreak in order to make a complete clinical and analytical assessment of the disease caused by the pathogen in this species. Viral load (by real-time RT-PCR) in blood, oral and cloacal swabs, and feathers, and neutralizing antibody titres (by VNT) were measured. In order to determine direct contact transmission, non-inoculated partridges were caged together with the inoculated ones. To assess infectiousness in other species, house sparrows and mice were also inoculated with the virus. All the inoculated partridges were clinically affected, and 30% of them died. All the infected individuals lost weight, with larger losses being recorded in females. Conversely, no mortality or disease symptoms were observed in the sparrows or mice. Remarkably, all the contact partridges acquired the infection by direct (non-vectored) transmission. This study confirms that the red-legged partridge is a susceptible host for BAGV infection, and that this pathogen is transmitted by direct contact. Long-lasting viral loads detected in calami of immature feathers demonstrate that feather sampling could be a useful strategy in active surveillance programs for early detection of BAGV.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-015-0233-9) contains supplementary material, which is available to authorized users.     

West Nile virus-associated mortality events in domestic Chukar partridges (Alectoris chukar) and domestic Impeyan pheasants (Lophophorus impeyanus)

West Nile virus (WNV) infection was diagnosed in captive juvenile chukars (Alectoris chukar), and captive juvenile Impeyan pheasants (Lophophorus impeyanus) on the basis of necropsy, histopathology, polymerase chain reaction, and immunohistochemistry. The chukars were kept in a game bird farm that experienced two outbreaks with approximately 25% mortality in hundreds of chukars between September and October 2002 and during the same months in 2003. The submitted pheasants were part of a group of 15 juvenile Impeyan pheasants that all died within approximately 2 wk at the end of August 2002. The macroscopic lesions in the pheasants were dominated by mucosal hemorrhage at the proventricular to ventricular junction and cecal ulcers, whereas the gross lesions in the chukar partridges were nonspecific. The predominant microscopic lesion in the chukar partridges was myocardial necrosis, whereas fibrinous and necrotizing splenitis was prominent in the pheasants. Viral antigen was usually widespread in animals of both species. Spontaneously occurring WNV infection should be considered a differential diagnosis in cases of mortality among select species of galliform birds.     

Pathogenesis of marble spleen disease in bursectomized and non-bursectomized ring-necked pheasants following oral inoculation with cell-culture-propagated virus.

Seventy-two 13-week-old ring-necked pheasants were inoculated orally with 5.0 x 10(2) tissue-culture infective dose (TCID) of cell-culture-propagated marble spleen disease virus. Inoculated birds exhibited neither mortality nor clinical disease. Gross and histologic lesions were typical of marble spleen disease. The mean splenic weight was significantly (P less than 0.02) higher in inoculated birds than in controls between 6 and 10 days postinoculation (PI). The histologic splenic lesions, which consisted of reticuloendothelial cell hyperplasia, intranuclear inclusions within reticuloendothelial cells, and lymphoid depletion, were most prominent between 6 and 10 days PI. In a second experiment, 1-day-old pheasants were chemically bursectomized by dosing birds with 1.2 mg cyclophosphamide on 3 consecutive days. At 7 weeks of age, 54 bursectomized birds were inoculated orally with 5.0 x 10(2) TCID of marble spleen disease virus. Gross and histologic lesions were detected in one of the inoculated pheasants, but the mean splenic weight was not significantly different from control birds at any time PI. These results are evidence of the role of the bursa of Fabricius in the pathogenesis of marble spleen disease.     

Pathogenesis and transmissibility of highly (H7N1) and low (H7N9) pathogenic avian influenza virus infection in red-legged partridge (Alectoris rufa)

ABSTRACT: An experimental infection with highly pathogenic avian influenza virus (HPAIV) and low pathogenic avian influenza virus (LPAIV) was carried out in red-legged partridges (Alectoris rufa) in order to study clinical signs, gross and microscopic lesions, and viral distribution in tissues and viral shedding. Birds were infected with a HPAIV subtype H7N1 (A/Chicken/Italy/5093/1999) and a LPAIV subtype H7N9 (A/Anas crecca/Spain/1460/2008). Uninoculated birds were included as contacts in both groups. In HPAIV infected birds, the first clinical signs were observed at 3 dpi, and mortality started at 4 dpi, reaching 100% at 8 dpi. The presence of viral antigen in tissues and viral shedding were confirmed by immunohistochemistry and quantitative real time RT-PCR (qRRT-PCR), respectively, in all birds infected with HPAIV. However, neither clinical signs nor histopathological findings were observed in LPAIV infected partridges. In addition, only short-term viral shedding together with seroconversion was detected in some LPAIV inoculated animals. The present study demonstrates that the red-legged partridge is highly susceptible to the H7N1 HPAIV strain, causing severe disease, mortality and abundant viral shedding and thus contributing to the spread of a potential local outbreak of this virus. In contrast, our results concerning H7N9 LPAIV suggest that the red-legged partridge is not a reservoir species for this virus.     

Some observations on the response of ring-necked pheasants to inoculation with various strains of cell-culture-propagated type II avian adenovirus

The response of ring-necked pheasants to inoculation with three strains of cell-culture-propagated type II avian adenovirus was examined. Marble spleen disease (MSD) virus of pheasants and both avirulent and virulent strains of hemorrhagic enteritis virus (HEV) of turkeys all induced typical gross and microscopic splenic lesions of MSD; neither MSD-associated lung lesions nor mortality were noted in inoculated pheasants, regardless of strain of virus used. Pheasants inoculated with a cell-culture-propagated avirulent strain of HEV were properly immunized against challenge with virulent HEV, as indicated by seroconversion and by protection against virus-induced splenic lesions. We conclude that these strains of type II avian adenovirus are comparable in pathogenicity for pheasants and cannot be distinguished. Further, absence of MSD-associated lung lesions and mortality in pheasants maintained under controlled laboratory conditions suggest that other environmental factors are probably involved in induction of such lesions and mortality in field cases of MSD.     

An outbreak of fowl cholera in ring-necked pheasants (Phasianus colchicus)

A total of 120 ring-necked pheasants from a 3000-bird flock in Zeeland, MI, died over a 3-day period. Clinical signs included sudden death, diarrhea, and limping. At necropsy, hepatomegaly with multifocal cream-colored foci randomly distributed throughout the parenchyma was observed in diseased birds. Additionally, the spleen was enlarged up to three times its normal size and had a marbled appearance. Microscopically, there was multifocal splenic and hepatic necrosis with intralesional rod-shaped bacteria. Pasteurlla multocida serotype 3/4 was isolated from liver and spleen. In this paper, we report an outbreak of acute fowl cholera in ring-necked pheasants.     

禽流感病毒番鸭分离株对不同禽类致病性的研究

将禽流感病毒番鸭分离株ZM的感染性鸡胚尿囊液静脉或肌肉接种不同禽类,同时设鸡源JC株对照,以了解毒株的致病特点。结果显示ZM株与同期分离JC株致病能力相当,两毒株均对鸡、鹅、番鸭、鹌鹑和鹧鸪显示了很强的致病能力,致死率50％-100％,死亡禽类的肝、肾、肺、脾、胰腺等多个组织器官均存在严重的损伤。鸽子,肉鸭和绍鸭(除zM株致死一只绍鸭外)均未发生死亡,外观健康。     

Pathobiology of A/chicken/Hong Kong/220/97 (H5N1) avian influenza virus in seven gallinaceous species

Direct bird-to-human transmission, with the production of severe respiratory disease and human mortality, is unique to the Hong Kong-origin H5N1 highly pathogenic avian influenza (HPAI) virus, which was originally isolated from a disease outbreak in chickens. The pathobiology of the A/chicken/Hong Kong/220/97 (H5N1) (HK/220) HPAI virus was investigated in chickens, turkeys, Japanese and Bobwhite quail, guinea fowl, pheasants, and partridges, where it produced 75-100% mortality within 10 days. Depression, mucoid diarrhea, and neurologic dysfunction were common clinical manifestations of disease. Grossly, the most severe and consistent lesions included splenomegaly, pulmonary edema and congestion, and hemorrhages in enteric lymphoid areas, on serosal surfaces, and in skeletal muscle. Histologic lesions were observed in multiple organs and were characterized by exudation, hemorrhage, necrosis, inflammation, or a combination of these features. The lung, heart, brain, spleen, and adrenal glands were the most consistently affected, and viral antigen was most often detected by immunohistochemistry in the parenchyma of these organs. The pathogenesis of infection with the HK/220 HPAI virus in these species was twofold. Early mortality occurring at 1-2 days postinoculation (DPI) corresponded to severe pulmonary edema and congestion and virus localization within the vascular endothelium. Mortality occurring after 2 DPI was related to systemic biochemical imbalance, multiorgan failure, or a combination of these factors. The pathobiologic features were analogous to those experimentally induced with other HPAI viruses in domestic poultry.     

Immune function in pheasants experimentally infected with marble spleen disease virus.

Two experiments were conducted to evaluate the effect of marble spleen disease virus (MSDV) infection on the immune response of pheasants. In the first, 15 ring-necked pheasants were inoculated orally with cell-culture-propagated MSDV and 15 received saline (controls). On days 7, 21, and 35 postinoculation (PI), all birds received sheep erythrocytes intravenously. Hemagglutination titers to sheep erythrocytes were determined for serum samples collected weekly for 6 weeks. The virus-inoculated group had significantly (P less than 0.05) lower hemagglutination titers than the control group. In the second experiment, 30 pheasants were allotted into two groups as above. Whole blood was collected from each bird weekly for 5 weeks. The blood was cultured in microtiter plates with or without optimum concentrations of concanavalin A. Five of 10 MSDV-inoculated pheasants had significantly depressed T lymphocyte transformation on either day 7 or day 14 PI. Overall, the depression of T lymphocyte transformation was transient and mild.     

Pulmonary congestion and edema (marble spleen disease) of chickens produced by group II avian adenovirus

"Marble spleen disease" of chickens was diagnosed in 22-week-old chickens. Total mortality was 8.9%. Deaths occurred over a period of 2 months. Gross lesions included pulmonary congestion, splenomegaly, hepatomegaly, and congestion of egg follicles. Microscopic lesions included pulmonary congestion and edema, and reticuloendothelial cell hyperplasia of the spleen with concurrent white-pulp necrosis and lymphocyte depletion. The pulmonary lesions were of sufficient intensity to have caused the death of fatally affected birds. Many of the hyperplastic reticuloendothelial cells contained basophilic intranuclear inclusions similar to those that characterize hemorrhagic enteritis of turkeys, marble spleen disease of pheasants, and adenovirus group II splenomegaly of chickens. These characteristic lesions, plus serologic identification of the causal virus, indicate that "marble spleen disease" caused by avian group II adenovirus was affecting the flock under study. This appears to be the first report of death of chickens due to pulmonary congestion and edema caused by spontaneous infection with avian group II adenovirus.     

Detection of Mycoplasma synoviae in clinically normal pheasants

Mycoplasma synoviae was isolated from the tracheas of seven clinically normal pheasants found in the vicinity of a chicken farm infected with M synoviae, but not from 120 pheasants and partridges with respiratory disease. When specimens were examined by the polymerase chain reaction only two additional pheasants infected with M synoviae were identified, one healthy and one diseased.     

An unusual case of coccidiosis in laboratory-reared pheasants resulting from a breach in biosecurity

An outbreak of coccidiosis in laboratory-reared Chinese ring-necked pheasants (Phasianus colchicus) resulted in high morbidity and moderate mortality. The outbreak was associated with a breach in biosecurity caused by the cleaning of a sewer line with a mechanical device, resulting in extensive splattering of fecal material throughout the "clean room" where birds were held prior to use in coccidiosis experiments. Mortality and morbidity in the affected birds were seen exactly 5 days after the incident, after birds had been moved to another room for experimental use, corresponding closely with the known prepatent or preclinical period of Eimeria phasiani and Eimeria colchici. Gross lesions in the affected birds varied from dehydration to intestinal and ventricular hemorrhage. Microscopic examination confirmed a diagnosis of severe intestinal coccidiosis. This report underscores the ease of contamination of experimental birds leading to coccidiosis outbreaks during breaches of management and biosecurity.     

Isolation from predators: a key factor in the failed release of farmed red-legged partridges (Alectoris rufa) to the wild?

1. The effects of predation isolation were studied in two lines of 10 month-old farmed red-legged partridges (Alectoris rufa). Birds came from bred generations in captivity (F) and directly from wild parents (W). 2. Releases were carried out in March for two consecutive years on private farmland with a stable population of wild red-legged partridge using 68 tagged birds (36 F, 32 W). 3. No statistical differences were found in survival time (mean 18 d) or home range values (mean 7·38 ha); 49% of mortality was attributable to raptors, 36% to wild canids and 15% had doubtful causes of death. F birds suffered more predation by raptors and W partridges more by mammalian carnivores. 4. A statistical difference was found in post-release dispersion: the means were 243·4 m in F and 593·3 m in W. 5. The result of predation isolation was similar in both groups, which confirms the adverse effects of current production systems on anti-predator behaviour. Farmers should consider the introduction of new rearing techniques with the aim of improving survival after release.     

The influence of colostrum on infection of calves around 7 months of age with Schistosoma mattheei

Thirty red-legged partridges (Alectoris rufa), 5-month-old, were orally inoculated with oocysts of the OV-51/95 strain of Toxoplasma gondii. Birds were distributed into five groups and received, respectively, 10 (group A, 4 birds), 50 (group B, 14 birds), 10(2) (group C, 4 birds), 10(3) (group D, 4 birds) and 10(4) (group E, 4 birds) oocysts. One partridge from group B and one from group E died suddenly of acute toxoplasmosis at 7 day after inoculation (DAI) with demonstrable T. gondii in several tissues. The rest of birds remained clinically normal until killed at 44, 58, 65, 72, 79 or 100 DAI. Brain, heart, liver and skeletal muscle from these partridges were bioassayed individually in mice; T. gondii was demonstrated in all these tissues, except in heart of three birds inoculated, respectively, with 10, 50 and 10(2) oocysts. Lesions were not seen in histologic sections of tissues from surviving partridges. These results suggest that red-legged partridges are resistant to clinical toxoplasmosis.     

Transmission of Histomonas meleagridis to domestic fowl by means of earthworms recovered from pheasant yard soil.

Four cockerels and two poults were fed ground earthworms recovered from soil where ring-necked pheasants (Phasianus colchicus) were raised. All 4 cockerels showed mild signs of histomoniasis, and numerous Heterakis were recovered at necropsy. One of the 2 poults died, with typical gross and microscopic lesions of histomoniasis. Heterakis eggs recovered from the 4 cockerels were used to infect an additional 12 cockerels and 11 poults.     

Large intestine in pheasant (phasianus colchicus) and see-see partridge (ammoperdix griseogularis). A comparative morphological and histological study

Anatomical and histological studies of the large intestine in birds are essential for necropsy and histopathological examinations. On the other hand, large intestine plays a critical role in the immune system and absorption. The present study's objective was to provide information about the morphological, morphometric and histological characteristics of the large intestine in see-see partridges (Ammoperdix griseogularis) and pheasants (Phasianus colchicus). These two members of the Galliformes order are found in large areas of Iran. In this study, ten male see-see partridges and ten male pheasants were randomly selected. Also, the histological studies were carried out on tissue specimens using haematoxylin for staining. In both species, the cecum was visible as two large tubes at the beginning of the rectum. The cecum and rectum's outer surfaces were flat and had no sacculation in both of the birds. Histologically, intestinal villi in all the large intestine parts and increase their height from the base to the apex is remarkable. The muscularis mucosae was distinct, and lymph nodes and Liberkuhn glands were found in all parts of the large intestine, approximately. It can be concluded that the morphology of the large intestine is very similar to the other avian species, but there are more differences in the histological features. These structural features are in full accordance with the bird's habits and diet.     

Efficacy of ionophorous anticoccidial drugs against coccidia in farm-reared pheasants (Phasianus colchicus) from Illinois

Litter samples obtained from a ring-necked pheasant propagation farm in Illinois contained coccidia: 57.5% of the oocysts were Eimeria duodenalis, 24.9% were E. tetartooimia, 8.8% were E. phasiani, and 8.8% were E. pacifica. Ionophorous anticoccidial drugs were tested for efficacy against the pheasant coccidia. All three drugs reduced oocyst production and prevented mortality in young pheasants; unmedicated infected controls had a 40% mortality rate. Monensin at 120 ppm in the feed was coccidiocidal against E. duodenalis and E. tetartooimia, partly coccidiocidal against E. pacifica, and only partly coccidiostatic against E. phasiani. Salinomycin at 60 ppm in the feed was highly efficacious and coccidiocidal against all four species, but the salinomycin-medicated pheasants gained the least of all medicated birds. Lasalocid at 120 ppm in the feed was the most effective, with nearly complete coccidiocidal activity against all four coccidial species.     

Prevalence of infectious diseases in ring-necked pheasant flocks in Poland

The health status of ring-necked pheasants in view of the prevalence of infectious diseases was estimated in Polish pheasantries in the years 1997-2000. Anatomicopathological, microbiological and serological examinations were carried out on birds derived from 26 pheasantries, including birds randomly selected from 18 flocks and sick or dead birds sent from 8 pheasantries. Antibodies specific to the following viruses were detected in serum blood samples: HE, AE, AP, REO, AI, Adeno group 1, MD, ND, as well as Mycoplasma gallisepticum specific antibodies. However, in none of the examined flocks was the presence of antibodies against reticuloendoteliosis virus found. Marble spleen disease and salmonellosis proved to be the most frequent cause of death during the growing period.     

A case of acute pulmonary edema, splenomegaly, and ascites in guinea fowl

Acute pulmonary edema, splenomegaly, and ascites were observed in a disease outbreak in adult white and pearl guinea fowl. The clinical history and gross and microscopic lesions resembled those described for marble spleen disease of pheasants and avian adenovirus group II splenomegaly of chickens. A small number of intranuclear inclusion bodies were found in liver, spleen, and lung sections of affected guinea fowl. Attempts to isolate virus and serological tests to detect the presence of viral antigens were unsuccessful. Adult female pearl guinea fowl experimentally exposed to pheasant and turkey isolates of type II avian adenoviruses developed gross and microscopic lesions similar to those seen in the field outbreak. The pheasant isolate was the more virulent. Intranuclear inclusion bodies were observed in liver, spleen, and lung sections of pearl guinea fowl inoculated with either of the virus isolates, and direct immunofluorescent examination revealed viral antigen in the spleen and lung.     

Marble Spleen Disease of Pheasants in Ontario

Two outbreaks of marble spleen disease were observed in pen-raised pheasants in Ontario on the same farm in the summer of 1971 and in 1972. In both instances, a mortality of 3% per day lasted for approximately 14 days. Fifteen birds were necropsied and tissues removed and examined histologically and by electron microscopy. In addition splenic material was injected into adult pheasants. The gross lesions, characterized by enlarged greyish-white mottled spleens and acute congestion and oedema in the lungs, were identical to those described by previous authors. Histological examination of livers, lungs, spleens and proventriculi revealed some reticular-type cells which contained either an eosinophilic or a basophilic intranuclear inclusion. Electron microscopy of the affected spleens revealed cells with massive intranuclear accumulations of virus particles. Although transmission trials failed, the evident histological and electron microscopy lesions confirm the suggestion by other authors that this is a viral disease affecting the reticulo-endothelial system.