Chicken proventricular necrosis virus (CPNV), isolate R11/3, previously was isolated from transmissible viral proventriculitis–affected chickens and was determined to be the likely etiology of this disease. CPNV was identified as a birnavirus on the basis of virion size and morphology (icosahedral, approximately 75 nm in diameter, nonenveloped); buoyant density in cesium chloride (1.32 g/ml); a genome comprising bisegmented, double-stranded RNA (approximately 3.8 and 3.4 kilobase pairs); and nucleotide sequence analyses. Nucleotide sequencing of CPNV RNA, segment B, identified a single large open reading frame that encodes a 903–amino acid protein. The 903–amino acid protein was identified as the putative VP1, the viral RNA-dependent RNA polymerase (RdRp), on the basis of sequence homologies with other birnavirus VP1 proteins. The CPNV VP1 possessed the unique permuted RdRp sequence motif arrangement characteristic of birnaviruses; however, phylogenetic analyses based on VP1 demonstrated that CPNV is deeply divergent from other birnaviruses.

Mycoplasma meleagridis (MM) is a major cause of disease and economic loss in turkeys. Formerly it was thought that this species was very host specific and only restricted to turkey. In this study, we report on the recovery of MM from breeding flocks of chickens located near a turkey breeding unit. Ten MM field strains were isolated (by culture on Frey broth medium) from tracheal swabs of chickens displaying clinical signs of mycoplasmosis—essentially respiratory symptoms and poor performance. Assignment of the isolated field strains to MM was confirmed by a growth inhibition assay using MM-specific polyclonal antiserum and by PCR amplification targeting the 16S rRNA sequence as well as the Mm14 sequence, a MM-species–specific DNA fragment previously identified and characterized in our laboratory. The nucleotide sequence of Mm14 proved to be highly conserved among the 10 MM field strains, indicating a common source of infection. However, on the basis of slight differences in sodium dodecyl sulfate–polyacrylamide gel electrophoresis whole-cell proteins and western blot profiles, two groups of the isolated MM field strains could be distinguished. Evidence of MM infection of chickens was further provided by serology, since 13.77% (35/254) of sera proved positive to MM by either rapid serum agglutination or recombinant antigen-based enzyme-linked immunosorbent assay. In addition, sera of all chickens from which MM was isolated were positive for antibodies to MM. Collectively, the data unambiguously show that MM could infect chickens; thus, MM warrants further exploration to determine its pathogenicity in this unusual host.

We analyzed the involvement of the rural poultry sector in outbreaks of low pathogenicity avian influenza (AI) in Italy in 2007–2009 and discuss possible measures for improving monitoring and control. A description of how the rural poultry sector is organized also is provided. Data were obtained by the AI surveillance system established in the areas affected by the outbreaks. The surveillance activities identified two H7N3 epidemics, in 2007 and 2009, both of which mainly involved the rural sector, yet these activities did not allow for the prompt eradication of the disease. Additional strategies could be adopted to avoid the persistence of AI within the rural sector, based on the regulation and control of poultry holdings at the top of the production chain.

Highly pathogenic avian influenza viruses of the subtype H5N1 (HPAIV-H5N1) have circulated continuously in Asia, Europe, and Africa since 2003. Investigations on the environmental preference and global spread processes of the virus are needed. We compiled 16 environmental variables to assess their correlation with HPAIV-H5N1 occurrences by using a niche-based model called Maxent. We found the virus had the strongest positive association with the human footprint index, as well as the presence of certain types of wetlands and mild temperature (10–30 C). Outbreaks of HPAIV-H5N1 in poultry or wild birds were also more frequent in certain major habitat types (e.g., tropical and subtropical moist broadleaf forests, temperate broadleaf and mixed forests, and flooded grasslands and savannas) and ecoregions. We conducted trend surface analysis to generate the travelling wave of the virus' global spread from 2003 to 2009, which indicated that high mountains or plateaus did not affect the spread speed and direction.

Four commercial strains of chickens, namely, ISA brown leghorn (ISA), TETRA-SL brown (TETRA-SL), Lohmann brown (LB), and Lohmann LSL (LSL), were infected with a well-defined clonal culture of Histomonas meleagridis (H. meleagridis/Turkey/Austria/2922-C6/04) to investigate their susceptibility to histomonosis. Each group included 16 chickens, which were housed under the same conditions in separate pens. All chickens were infected with 10⁴ histomonads orally and intracloacally at 14 days of age. No mortality or clinical signs were observed during the experiment in all birds. Three birds of each chicken strain were euthanatized on days 4, 7, 10, 14, and 21 postinfection. Incidence of histomonosis on the basis of cecal lesions was found to be 64.00% in TETRA-SL, 62.50% in LB, 53.12% in LSL, and 43.75% in ISA chickens. Fewer lesions were noticed in livers than in ceca, with an incidence of 15.62% in TETRA-SL, 9.37% in LB, and 3.12% in ISA chickens. No liver lesions were found in the LSL chickens. Statistical analysis revealed that there was no significant difference (P > 0.05) in susceptibility to experimental H. meleagridis infection based on cecal and liver involvement. Polymerase chain reaction (PCR) and immunohistochemistry were found to be reliable tools to confirm the presence of histomonads and changes in the ceca. However, some negative PCR results were recorded from the livers despite the presence of macroscopic lesions. Additionally, DNA of H. meleagridis was detected by PCR in a few of the lungs, but immunohistochemistry was negative. Nucleic acid of the protozoan parasite was not detected in samples from kidney, brain, spleen, or bursa of Fabricius. Altogether, the high susceptibility of commercial chicken lines to histomonosis could be demonstrated and characterized by severe lesions in the ceca and insignificant involvement of the liver, approaching a maximum on days 7–14 postinfection.

We formulate a stochastic, spatial, discrete-time model of viral “Susceptible, Exposed, Infectious, Recovered” animal epidemics and apply it to an avian influenza epidemic in Pennsylvania in 1983–84. Using weekly data for the number of newly infectious cases collected during the epidemic, we find estimates for the latent period of the virus and the values of two parameters within the transmission kernel of the model. These data are then jackknifed on a progressive weekly basis to show how our estimates can be applied to an ongoing epidemic to generate continually improving values of certain epidemic parameters.

We evaluated protection conferred by mucosal vaccination with replication-competent adenovirus-free recombinant adenovirus expressing a codon-optimized avian influenza (AI) H5 gene from A/turkey/WI/68 (AdTW68.H5_ck). Commercial, layer-type chicken groups were either singly vaccinated ocularly at 5 days of age, singly vaccinated via spray at 5 days of age, or ocularly primed at 5 days and ocularly boosted at 15 days of age. Only chickens primed and boosted via the ocular route developed AI systemic antibodies with maximum hemagglutination inhibition mean titers of 3.9 log₂ at 32 days of age. In contrast, single vaccination via the ocular or spray routes maintained an antibody status similar to unvaccinated controls. All chickens (16/16) subjected to ocular priming and boosting with AdTW68.H5_ck survived challenge with highly pathogenic AI virus A/chicken/Queretaro/14588-19/95 (H5N2). Single ocular vaccination resulted in 63% (10/16) of birds surviving the challenge followed by a 44% (7/16) survival of single-sprayed vaccinated birds. Birds vaccinated twice via the ocular route also showed significantly lower (P < 0.05) AI virus RNA concentrations in oropharyngeal swabs compared to unvaccinated–challenged controls.

The conditions under which infection with Histomonas meleagridis could spread from directly inoculated turkey poults to uninoculated poults without the aid of invertebrate hosts or vectors was investigated in several experiments. In three experiments in battery cages, uninoculated poults were commingled with directly infected birds on pine-shaving litter. Directly exposed birds were inoculated per cloaca with H. meleagridis by means of a plastic pipette tip attached to a 10-ml syringe or orally gavaged with fresh cecal droppings from donor turkeys 4 days postinoculation (PI). Of the cloacally inoculated controls in these experiments, 31 of 44 (70.5%) birds had severe lesions of histomoniasis at 14 days PI, whereas none of the orally gavaged birds became infected. Histomoniasis developed in 11 of 36 (30.5%) birds allowed to commingle with inoculated birds. In other treatments, poults were allowed only contact with droppings from directly inoculated birds after the infected birds were removed from the cages. This was done for a single period of 1 hr or repeated five times. Four of 32 birds (12.5%) became infected in this way after the single exposure, whereas only four of 44 birds (9.1%) exposed five times developed lesions. In a comparison of floor materials, 35 of 35 control birds inoculated per cloaca developed severe liver and cecal lesions, irrespective of litter. Uninoculated birds allowed to commingle with infected birds on paper or pine shavings became severely infected in all cases (12/12 and 12/12 birds, respectively), whereas only 33% of those on wire-floored cages became infected (4/12). These results suggest that transmission of infection is more likely to occur as a result of direct contact between birds than from contact with litter or fecal material.

High dietary protein is thought to increase the incidence of foot pad dermatitis (FPD) as a result of increased uric acid and secondary ammonia production in the excreta or litter. This study was conducted on female turkeys over a period of 3 wk to test the effects of water alone, and also of these end products of protein metabolism, independent of the presence of excreta, on the development and severity of FPD. The animals were allocated into four groups, with 20 birds in each, and housed in floor pens on dry, clean wood shavings (changed daily) throughout the experiment. The control group was housed continuously in its pen, whereas the other groups were additionally exposed daily (for 8 hr) to experimentally treated wood shavings, in adjacent separate boxes, enriched with water alone or water with NH₄Cl or uric acid. NH₄Cl and uric acid were added via water to the litter to achieve the concentrations of ammonia and uric acid in the litter as found in the excreta of turkeys (about 0.50 g ammonia and 20 g uric acid/kg). The wet litter was kept clean by removing the excreta twice daily and by changing the litter twice a week. The foot pads of all birds were examined on days 0, 7, 14, and 21 and scored externally (macroscopically). Three birds were selected from each group on days 0, 7, and 14, while the remaining 11 birds/group were sacrificed on day 21 for histopathologic assessment of foot pads. The severity of FPD was found to be markedly higher (about 3 times) on wet than on dry litter. There were no negative effects of ammonia and uric acid on foot pad lesions. The results indicate that high litter moisture is the most likely factor causing FPD in turkeys. A focus on nitrogenous irritants in the litter was not substantiated. Exposure of birds to wet litter (in the absence of excreta) for only 8 hr/day was sufficient to develop foot pad lesions. The present results suggest that a focus on the protein content of the diet as a cause of FPD may be misplaced, but all dietary factors which increase excreta or litter moisture, or both, should be considered. The prevalence and severity of FPD can be reduced by maintaining dry litter.

To determine whether chemotherapeutic compounds available for use in domestic poultry are effective at controlling coccidiosis in northern bobwhites (Colinus virginianus), we tested 13 chemotherapeutic anticoccidials including amprolium (250 parts per million [ppm]), clopidol (125 ppm), diclazuril (1 ppm and 2 ppm), decoquinate (30 ppm), lasalocid (120 ppm), monensin (90 ppm), narasin/nicarbazin (36/36 ppm), robenidine (33 ppm), roxarsone (50 ppm), sulfadimethoxine/ormetoprin (125/75 ppm), salinomycin (60 ppm), semduramicin (25 ppm), and zoalene (125 ppm and 150 ppm). Three tests were conducted using two replicates of 10 birds each: Infected, unmedicated controls and medicated birds were challenged with 1 × 10⁶ oocysts of a field isolate consisting primarily of Eimeria lettyae. Subsequently, we tested clopidol, lasalocid, salinomycin, diclazuril (1 ppm), and monensin against mixed-species field isolates containing E. lettyae, E. dispersa, E. colini, or all. Weight gain, gross intestinal lesions, severity of diarrhea, and feed conversion ratio (FCR) 6 days postinfection were recorded. Lesion score, as previously reported, was unreliable as a measure of severity of infection in comparison with weight gain, fecal scores, and FCR. Excellent to good efficacy was found in clopidol, decoquinate, diclazuril (1 ppm and 2 ppm), and in lasalocid, narasin and nicarbazin, robenidine, sulfadimethoxine/ormetoprin, and zoalene (150 ppm). Marginal protection was found using monensin, salinomycin, semduramicin, or a roxarsone/semduramicin combination. Amprolium, roxarsone, and zoalene (125 ppm) were ineffective at controlling coccidia. Two of the six isolates tested against diclazuril 1 ppm and clopidol demonstrated a high degree of resistance, but none of the six isolates was resistant to lasalocid. Four of the eight isolates showed mild to moderate, and moderate to high, resistance against monensin and salinomycin, respectively. These findings indicate that several available compounds are effective at controlling coccidiosis in bobwhites.

Avian nephritis virus (ANV), which belongs to the Astroviridae family, has been associated with acute nephritis in chickens. Cases of ANV infection have been recorded in Japan and in several European countries. However, related studies have never been performed in China. Thus, this study isolated ANV in Chinese chicken flocks. ANV RNA was detected by reverse transcription–PCR in stool samples collected from healthy layer chickens in the Sichuan Province of China in 2009. Of the 192 stool specimens collected, 32.3% (62/192) were positive for ANV infection. The whole genome of ANV-Sichuan54, the first representative Chinese strain, was 6941 nucleotides in length, including the 5′ untranslated region, three open reading frames (ORFs), a 3′ UTR, and a poly-(A) tail. Comparative and phylogenetic analyses based on partial RNA-dependent RNA polymerase (ORF1b) demonstrated that the majority of ANV investigations were more closely related to the U.S. ANV strain (DQ324827–324836) than to the G-4260 (AB033998).

A reverse-transcriptase–polymerase-chain-reaction (RT-PCR) procedure was evaluated for detection of chicken proventricular necrosis virus (CPNV) in transmissible viral proventriculitis (TVP) –affected chickens. The RT-PCR procedure was compared with indirect immunofluorescence (IFA) and virus isolation for detection of CPNV in experimentally infected chickens. Microscopic lesions characteristic of TVP were detected on days 5–35 postexposure (PE) in CPNV-infected chickens; CPNV was detected by RT-PCR on days 3–14 PE in freshly collected proventriculi, and on days 1–14 PE in formalin-fixed paraffin-embedded (FFPE) proventriculi. CPNV was detected in proventriculi of experimentally infected chickens by IFA on days 3–10 PE, and by virus isolation on days 1–14 PE. With IFA used as a reference, sensitivity of the RT-PCR procedure with freshly collected and FFPE proventriculi was 88% and 100%, respectively; specificity was 83% and 86%, respectively. Proventriculi (FFPE) obtained from suspect TVP cases (n  =  19) were evaluated for presence of CPNV by RT-PCR and microscopic lesions consistent with TVP. CPNV was detected by RT-PCR in proventriculi from 8/11 TVP ( ) cases (24/36 tissue sections). TVP ( ) cases were defined by microscopic lesions characteristic of TVP; CPNV was not detected in proventriculi (0/8 cases, 0/32 tissue sections) in the absence of these lesions. The association between presence of TVP-characteristic microscopic lesions and presence of CPNV was highly significant (P  =  0.0014). These findings indicate the utility of the RT-PCR procedure for detection of CPNV and provide additional evidence for an etiologic role for this virus in TVP.

The objective of this study was to isolate and identify suspected pathogens from peacocks and peacock farmers with severe pneumonia and to investigate its potential association with peacocks' pneumonia, caused by Chlamydophila psittaci infection. A clinical examination of infected peacocks identified birds with symptoms of anorexia, weight loss, yellowish droppings, airsacculitis, sinusitis, and conjunctivitis, whereas the infected farmers showed high fever and respiratory distress. Immunofluorescence tests detected chlamydial antigens in pharyngeal swabs (12 of 20) and lung tissue samples (four of five) from peacocks. One of four swabs taken from farmers was also positive by the same test. Specific anti-chlamydia immunoglobulin G was detected in 16 of 20 peacocks and four of four peacock farmers. The isolated pathogen was able to grow in specific-pathogen-free (SPF) chicken embryos and McCoy cell lines and was identified as Chlamydiae by immunofluorescence assay and PCR. Avian influenza virus, Newcastle disease virus, and infectious bronchitis virus were eliminated as potential causative agents after pharyngeal swabs inoculated onto the chorioallantoic membrane of embryonate eggs failed to recover viable virus. PCR and restriction fragment length polymorphism indicated the ompA gene from the isolate was similar to that of avian C. psittaci type B. Three-week-old SPF chickens challenged with the peacock isolate via intraperitoneal injection showed a typical pneumonia, airsacculitis, and splenitis. Subsequently, the inoculating strain was recovered from the lungs of challenged birds. This is the first report of C. psittaci infection in peacocks and peacock farmers.

Avian metapneumovirus (AMPV) causes turkey rhinotracheitis and is associated with swollen head syndrome in chickens, which is usually accompanied by secondary infections that increase mortality. AMPVs circulating in Brazilian vaccinated and nonvaccinated commercial chicken and turkey farms were detected using a universal reverse transcriptase (RT)-PCR assay that can detect the four recognized subtypes of AMPV. The AMPV status of 228 farms with respiratory and reproductive disturbances was investigated. AMPV was detected in broiler, hen, breeder, and turkey farms from six different geographic regions of Brazil. The detected viruses were subtyped using a nested RT-PCR assay and sequence analysis of the G gene. Only subtypes A and B were detected in both vaccinated and nonvaccinated farms. AMPV-A and AMPV-B were detected in 15 and 23 farms, respectively, while both subtypes were simultaneously found in one hen farm. Both vaccine and field viruses were detected in nonvaccinated farms. In five cases, the detected subtype was different than the vaccine subtype. Field subtype B virus was detected mainly during the final years of the survey period. These viruses showed high molecular similarity (more than 96% nucleotide similarity) among themselves and formed a unique phylogenetic group, suggesting that they may have originated from a common strain. These results demonstrate the cocirculation of subtypes A and B in Brazilian commercial farms.

Infection of maternal, antibody-negative chickens with chicken infectious anemia virus (CIAV) can cause clinical disease, while infection after maternal antibodies wane often results in subclinical infection and immunosuppression. Currently, vaccines are not available for vaccination in ovo or in newly hatched chickens. Development of CIAV vaccines for in ovo use depends on the ability to generate vaccines that do not cause lesions in newly hatched chicks and that can induce an immune response regardless of maternal immunity. Immune complex (IC) vaccines have been successfully used for control of infectious bursal disease, and we used a similar approach to determine if an IC vaccine is feasible for CIAV. Immune complexes were prepared that consisted of 0.1 ml containing 10^5.4 tissue culture infective dose 50% of CIA-1 and 0.1 ml containing 10 to 160 neutralizing units (IC Positive [ICP]10 to ICP160), in which one neutralizing unit is the reciprocal of the serum dilution required to protect 50% of CU147 cells from the cytopathic effects caused by CIA-1. Virus replication was delayed comparing ICP80 and ICP160 with combinations using negative serum (IC Negative [ICN]80 or ICN160). In addition, the number of birds with hematocrit values <28% were decreased with ICP80 or ICP160 compared to ICN80 or ICN160. Seroconversion was delayed in ICP80 and ICP160 groups. To determine if ICP80 or ICN160 protected against challenge, we vaccinated maternal, antibody-free birds at 1 day of age and challenged at 2 wk or 3 wk of age with the 01-4201 strain. Both ICP80 and ICP160 protected against replication of the challenge virus, which was measured using differential quantitative PCR with primers distinguishing between the two isolates. Thus, in principle, immune complex vaccines may offer a method to protect newly hatched chicks against challenge with field virus. However, additional studies using maternal, antibody-positive chicks in combination with in ovo vaccination will be needed to determine if immune complex vaccines will be useful to protect commercial chickens.

To determine the distribution of duck plague virus (DPV) gE protein in paraformaldehyde-fixed, paraffin-embedded tissues of experimentally DPV-infected ducks, an indirect immunoperoxidase assay was established to detect glycoprotein E (gE) protein for the first time. The rabbit anti-His-gE serum, raised against the recombinant His-gE fusion protein expressed in Escherichia coli BL21 (DE3), was prepared and purified. Western blotting and indirect immunofluorescence analysis showed that the anti-His-gE serum had a high level of reactivity and specificity and could be used as the first antibody for further experiments to study the distribution of DPV gE protein in DPV-infected tissues. A number of DPV gE proteins were distributed in the bursa of Fabricius, thymus, spleen, liver, esophagus, duodenum, jejunum, ileum, and kidney of DPV-infected ducks and a few DPV gE were distributed in the Harders glands, myocardium, cerebrum, and lung, whereas the gE was not seen in the skin, muscle, and pancreas. Moreover, DPV gE was expressed abundantly in the cytoplasm of lymphocytes, reticulum cells, macrophages, epithelial cells, and hepatocytes. The present study may be useful not only for describing the characteristics of gE expression and distribution in infected ducks but also for understanding the pathogenesis of DPV.

As no data are available on the prevalence of cytolethal distending toxin (cdt) genes carried by Campylobacter spp. in laying hens, this study was conducted with the aim to evaluate the prevalence of both Campylobacter spp. and cdt genes in 1680 laying hens from four different farms. The samples were analyzed by culture methods and by polymerase chain reaction. Campylobacter spp. were isolated from 1097/1680 cloacal swabs. Among the isolates, 913 were identified as Campylobacter jejuni whereas 345 were identified as Campylobacter coli. All isolates carried cdt genes. The results presented here confirm the very common occurrence of C. jejuni and C. coli in laying hens and underline that the cdt genes may also be frequently present in both C. jejuni and C. coli isolates from laying hens.

To assess the susceptibility of pigeons (Columba livia) to infection with H5N1 high pathogenicity avian influenza virus (HPAIV), four groups of 1-yr-old and 4-wk-old racing pigeons (10 birds in each group) were inoculated oculonasally with 10⁶ 50% egg infectious dose (EID₅₀) of A/crested eagle/Belgium/01/2004 (clade 1) or A/swan/Poland/305-135V08/2006 (clade 2.2). Contact specific-pathogen-free (SPF) chickens were kept in the same isolators as young pigeons (two chickens per group). At 3, 5, 7, 10, and 14 days postinfection (PI) two pigeons from each infected group were selected randomly, and oropharyngeal and cloacal swabs (pigeons and contact chickens) as well as a number of internal organs (pigeons) were collected for viral RNA detection in real-time reverse transcription PCR (RRT-PCR) and histopathology. At the end of the experiment (14 days PI) blood samples from two pigeons in each group and from contact SPF chickens were also collected, and sera were tested using hemagglutination inhibition (HI) test and blocking enzyme-linked immunosorbent assay (bELISA). During the observation period all pigeons remained clinically healthy, and no gross lesions were observed in any of the infected groups. SPF contact chickens were also healthy and negative in RRT-PCR and HI tests. However, the clade 1 H5N1 virus produced more sustained infection manifested by the presence of histopathologic changes (consisting mainly of mild to moderate hemorrhagic and inflammatory lesions), prolonged persistence of viral RNA (detectable between 3 and 10 days PI) in a variety of tissues of both adult and juvenile birds (with highest RNA load in lungs and brain) as well as slight viral shedding from the trachea and cloaca, but without transmission to SPF contact chickens. Additionally, two clade 1–infected adult pigeons sacrificed at the end of experiment showed seroconversion in bELISA and HI test (using homologous virus as antigen). The viral RNA was found only at day 3 PI in one adult pigeon inoculated with clade 2.2 H5N1 virus, but neither microscopic lesions nor seroconversion were found in any other tested birds inoculated with A/swan/Poland/305-135V08/2006. Our results support the observations that pigeons are resistant to H5N1 HPAIV (no deaths or clinical signs), but there may be clade-dependent differences in the pathogenic potentials of H5N1 HPAIV of Asian origin.

Mixtures of turkey herpesvirus (HVT) and Rispens poultry vaccines have been used worldwide for over 20 yr, mainly for vaccination of future layers and breeders. With increasing virulence of Marek's disease (MD) virus strains, vaccination strategies are evolving toward the use of vaccines combining HVT and Rispens. A single vaccination either in ovo or at 1 day of age with the HVT infectious bursal disease (IBD) vector vaccine is efficient against IBD. However, with vaccination programs that include a hatchery administration of the HVT IBD vaccine, additional protection against very virulent and very virulent–plus MD viruses is needed, especially for layers and breeders. This study looked at the combination of four commercially available Rispens vaccines with the HVT IBD vector vaccine injected at 1 day of age. MD challenge tests that were superior to 90% in relative score in all the groups vaccinated with both vaccines showed that the mixture of HVT IBD and Rispens vaccines had no effect on clinical protection against MD, and IBD challenge tests showed that the mixture of HVT IBD and Rispens vaccines had no effect on clinical protection against IBD, which was equal to 100% protection in all the groups vaccinated with both vaccines.

In order to evaluate the protection conferred by an experimental inactivated vaccine against infectious coryza, three challenge trials were undertaken using 112 1-day-old broilers. The vaccine “Hepa Inmuno NC” included bacterial antigens of Avibacterium paragallinarum (serogroups A, B, variant B, and C) as well as antigens of Newcastle virus and hepatitis virus. Fifty-six broiler chicks were vaccinated at the first day of life at the hatchery while another 56 chicks were left unvaccinated. Three infection trials were conducted simultaneously using each of the three serogroups A, B, or C of Av. paragallinarum. In each trial, 17 vaccinated and 17 unvaccinated broilers were used. Challenge was performed at day 31 of life by injection, into the left infraorbital sinus, of approximately 1 × 10⁵ colony forming units of the corresponding Av. paragallinarum strain. Clinical signs were recorded on day 2 postchallenge. All broilers were euthanatized and both infraorbital sinuses were bacteriologically examined for the presence of Av. paragallinarum on day 5 postchallenge. In comparison with the unvaccinated broilers, the vaccine significantly reduced the number of broilers with clinical signs after challenge with serogroup B, and significantly fewer vaccinated broilers were positive for the presence of Av. paragallinarum after challenge with serogroup C. On the other hand, no significant protection was observed when broilers were challenged with Av. paragallinarum from serogroup A. Despite the high infection rates in vaccinated chicks after an experimental infection with Av. paragallinarum, it was possible to reduce colonization of Av. paragallinarum (serogroup B) and clinical signs (serogroup C) in broiler chicks by vaccination at the first day of life. Further cross-protection trials should be done, including other Av. paragallinarum strains in the vaccine, especially those from serogroup A.

Rotavirus is a common pathogen causing gastroenteritis in humans and domesticated animals. The incidence of rotavirus in wild-living animals, particularly in avian species, has not been systematically investigated. In this study 1220 fecal samples and cloacal swabs collected from wild-living birds during 2008 in Hungary were tested for the presence of group A rotaviruses by a VP6 gene-specific reverse-transcription–polymerase-chain-reaction assay. Of the 1220 samples, 276 and 944 were processed as individual and pooled specimens, respectively. Rotavirus was identified in two pooled pheasant (Phasianus colchicus) samples and two individual reed bunting samples (Emberiza schoeniclus). These data indicated a very low prevalence of group A rotaviruses (0.3%) in our sample set. Nonetheless, the present study, together with existing literature data, implies that rotavirus infections occur in a wide spectrum of feral bird species. These findings are exciting and suggest that pursuing rotavirus monitoring is needed to uncover avian rotavirus strain diversity and understand rotavirus ecology in nature.

An adult red-legged partridge (Alectoris rufa) presented with concurrent pulmonary carcinoma and severe silicosis. The animal was submitted to the Veterinary Teaching Hospital of the University of Córdoba (Spain) because of respiratory signs, and it died during clinical examination. At postmortem examination, numerous firm, whitish to yellowish nodules involving the lungs, mainly the right lobe, were found. The histopathologic study revealed numerous peribronchiolar large granulomatous lesions composed of macrophages, which showed abundant cytoplasm containing numerous birefringent crystals identified as silicates by transmission electron microscopy. An epithelial neoplasm showing papillary, acinar, and solid patterns occupied large areas of the pulmonary parenchyma. The histopathologic and immunohistochemical features were consistent with a pulmonary carcinoma. Small tumor nests were often located close to the granulomatous lesions. This is the first report of concurrent pneumoconiosis and pulmonary carcinoma in a nonhuman species.

This report describes a case of dysgerminoma in a 21-year-old eastern rosella (Platycercus eximius eximius) that presented with dyspnea and a severely distended coelom. The bird was euthanatized, and a large, left-sided coelomic mass was identified. Microscopically, the mass was composed of sheets and nests of round to polygonal neoplastic cells with lacy cytoplasm. The neoplastic cells were weakly positive for vimentin and c-kit but negative for pancytokeratin, AE1, and inhibin. On the basis of the histomorphology and immunoreactivity, the neoplasm was determined to be a dysgerminoma. The variability of histologic appearance and immunohistochemical staining of dysgerminomas in humans compared with veterinary species is discussed.

A multiple-age egg production farm was conceived, designed, constructed, and managed with the goal of blocking the introduction of preventable poultry diseases and infestations. Fifteen years after conception and 13 years after housing the first hens, the 4 million-hen farm remains free of Mycoplasma gallisepticum, infectious laryngotracheitis, and Ornithonyssus sylviarum. Annual savings in vaccine use exceed the estimated annual costs of the biosecurity program. The cumulative design, construction, and operation of this farm are a successful demonstration of practical, effective biosecurity on a commercial poultry farm.

Avian pox viruses (APVs) have been reported to cause infection in diverse avian species worldwide. Herein we report the first case of APV infection in a free-living bird, a subadult crested serpent eagle (Spilornis cheela), in Taiwan. In addition to the typical wart-like lesions distributed on the cere, eyelid, and face, there were also yellowish nodules below the tongue and on the hard palate. Phylogenetic analysis of the 4b core protein gene showed that the APV is very close to that found in white-tailed sea eagle (Haliaeetus albicilla) in Japan recently. Because both cases are located on the same major flyway for migratory birds, the impact of this virus with regard to the wild and migratory raptor species along the East Asian–Australasian Flyway and West Pacific Flyway requires immediate investigation.

A common seagull (Larus fuscus) was found near the southern coast of Italy by the veterinarians of the local wild animal rescue center. Physical examination of the bird revealed an ulcerated mass involving a majority of the oral cavity; the mass did not allow for normal feeding. After the bird died necropsy was performed and the mass was histologically and immunohistochemically examined. The morphology and the immunoreactivity for Melan-A and S-100 antigens led to a diagnosis of malignant melanoma. This is the first case of malignant melanoma described in a seagull, and herein we compare the characteristics of the present case with malignant melanoma already described in domestic animals.

An outbreak of ascaridiasis occurred in 10-wk-old guinea fowl (Numida meleagris) on a commercial farm. Birds had exhibited elevated mortality (11.66%) in the previous week, as well as increased water consumption, weakness, anorexia, and stunted growth. Numerous nematodes, occasionally occluding the intestinal lumen, were present in the jejunum and ileum and were identified as Ascaridia numidae based on microscopic morphology. Ribosomal DNA 18S and 28S D3 sequences of the nematode were deposited into GenBank and found to be most similar to Ascaridia galli and Toxocara vitulorum, respectively; sequences for A. numidae had not been previously reported. Treatment with piperazine sulfate significantly reduced the number of adult worms in the intestines, greatly decreased eggs per gram of feces, relieved clinical signs in the flock, and returned the flock mortality back to expected levels. All findings implicate A. numidae as the cause of elevated mortality in this flock.

Fourteen infectious bronchitis viruses (IBVs) detected in broiler, broiler breeder, and layer flocks in Slovenia between 2007 and 2009 were molecularly analyzed by reverse transcription–PCR and direct sequencing of the S1 gene. Phylogenetic analyses based on partial S1 gene sequences revealed that seven strains were classified as the Italy 02 genotype, a genetic group of IBV that has not previously been detected in Slovenia. Another seven strains were classified as the QX genotype. The results of phylogenetic analyses and epidemiologic investigations revealed that the genetic classification correlates with the geographic origins of the IBV strains. Greater genetic variability (maximum nucleotide and amino acid divergences were 4.8% and 9.9%, respectively) was observed within the Slovenian strains from the Italy 02 genotype detected between 2007 and 2009 than within strains from the QX genotype isolated in 2008 and 2009 (1.2% and 1.3%, respectively). The Slovenian strains classified as the Italy 02 genotype form a separate genetic cluster. These strains shared five specific amino acid substitutions that became fixed during the period of surveillance. Strains from the QX genotype that shared one specific amino acid substitution also form a separate genetic cluster.