Circovirus infections have been documented in adult and nestling canaries (Fringillidae) but the distribution of the virus in the world is not yet known. In captive canary flocks, Circovirus infections have been reported based on the clinical observations. In this study, the presence of both canary circovirus (CaCV) and chicken anemia virus (CAV) in canary flocks was investigated. Virus strains were detected by PCR and direct sequencing of amplified products. Nucleotide sequences were aligned and compared with existing data in GenBank. PCR identified CaCV-positive birds, giving an overall positivity rate of 25%, but all samples were negative for CAV. According to the sequencing data, three distinct strains were identified. Our results indicated a relationship between genetic variation in the replicase gene (rep) and the geographic regions as well as the feasibility of using the rep gene for virus detection and molecular epidemiology investigations. We are reporting detection and characterization of canary circovirus based on the rep gene. Sequencing results and sequence identity analysis revealed that the rep gene could be used for detecting and discriminating the members of family Circoviridae. This manuscript is the first report of canary circovirus in Iran and of three new strains in the world.

In an effort to produce more stable vaccines for infectious laryngotracheitis virus (ILTV), recombinant strains with deletion of genes associated with virulence have been evaluated for attenuation and protection efficacy. Among viral genes associated with virulence, a cluster of five open reading frames (ORFs; A through E) have been identified. An attenuated ILTV recombinant strain with deletion of the ORF C gene induced protection comparable to that elicited by the tissue culture origin (TCO) vaccine when administered via eyedrop. The objective of this study was to evaluate the attenuation and protection efficacy of the ΔORF C strain when delivered in ovo to maternal antibody negative (MAb−) and maternal antibody positive (MAb ) embryos. In ovo delivery of the ΔORF C strain did not affected hatchability or body weight gain, while virus transmission to contact chickens was minor. Nevertheless, nine of ninety (10%) of MAb− chickens vaccinated with the ΔORF C strain showed marked dyspnea, and upon postmortem examination bloody mucoid plugs and high viral genome load were detected in their tracheas. Moreover, the ΔORF C strain induced satisfactory protection in MAb− chickens, but marginal protection in MAb chickens after challenge. The reduced protection observed for MAb groups of chickens was likely caused by the interference of maternally derived antibodies. This report presents the use of a genetically attenuated ILTV strain delivered in ovo as a potential new approach in the control of ILTV.

Streptococcal bacterial species represent common inhabitants of the intestinal tract of animals and humans with a potential for opportunistic infections. Streptococcosis has been identified in turkey poults (Meleagris gallopavo), ducklings and goslings (Anatidae), broiler chickens, semimature-adult chickens (Gallus gallus domesticus), and young and adult pigeons (Columbidae). However, the exact underlying factors that lead to bacterial invasion of the blood stream and tissue colonization have not been completely elucidated. The electronic database of the California Animal Health and Food Safety laboratory (Fresno, Tulare, and Turlock branches) was searched for necropsy cases in which streptococcosis was diagnosed in different avian species between January 2000 and August 2017. A total of 95 cases, involving both commercial operations and noncommercial premises, were analyzed. Streptococcus spp., Streptococcus bovis, and Streptococcus gallolyticus were identified from multiple organs, with macroscopic or histopathologic lesions (or both) indicative of septicemia in 23 (24%), 40 (42%), and 30 (32%) cases, respectively. Streptococcus pluranimalium and Streptococcus lutetiensis were also isolated from one (1%) and two (2%) cases, respectively. Turkey poults, broiler chickens, and ducklings were the most-commonly affected species with streptococcosis. Splenitis and hepatitis were the most-common lesions observed and these were the organs with the highest isolation rate. An overview of the clinical and pathologic presentation, and possible predisposing conditions associated with this bacterial infection, is provided.

Phosphorylated histone 3 (PH3) and cleaved caspase 3 (CCASP3) were used to detect proliferating and apoptotic cells, respectively, in the jejunums of female sibling poults, with and without enteritis and depressed growth, from hatch to day 35. Poults that developed enteritis and depressed growth (SIB flock) were raised on a commercial farm in eastern North Carolina, whereas poults with normal growth and no enteritis (TAU flock) were raised in the Teaching Animal Unit at North Carolina State University College of Veterinary Medicine. Beginning on day 5 through day 35 and at processing, TAU poults were significantly heavier than SIB poults. Jejunal weights, relative jejunal weights, and jejunal densities were greater in SIB poults from day 10 through 35. Jejunal efficiency (body weight /jejunal length) was higher in TAU poults at day 5 and days 10 through 35. Mucosal thickness was greater in SIB poults between days 7 and 21 but greater in TAU poults at days 28 and 35. From day 7 to 35, villus-to-crypt ratios were higher for TAU poults and lower for SIB poults because hyperplastic crypts formed a greater percentage of the mucosa in SIB poults. By day 7, PH3- and CCASP3-positive cells were increased in SIB poults, showing that mucosal changes resulted from combined crypt epithelial hyperplasia and increased apoptosis of villous enterocytes. Findings in this study confirm that enteritis, in the absence of clinical signs, and depressed growth in turkey poults begins by day 7, can be identified microscopically, persists for at least 35 days, is associated with lower processing weights, and has a profound negative effect on turkey growth.

The present study was performed to detect and characterize the serotypes of fowl adenovirus associated with inclusion body hepatitis (IBH) or hepatitis-hydropericardium syndrome (HHS) in commercial poultry in some regions of China between 2007 and 2017. Approximately 81 fowl adenovirus strains were isolated from liver or kidney samples from diseased poultry. A sequencing analysis of the hexon loop 1 gene revealed fowl adenovirus serotypes 8a, 8b, and 11 in samples of broilers with IBH, serotype 11 in layers with IBH, and serotype 4 in poultry with HHS. Of the fowl adenovirus serotype 4 strains, 62.07% were isolated from layers. Additionally, 74.07% of the isolated strains were fowl adenovirus serotype 11 prior to June 2014; 53.70% were serotype 4 after that time point; and strains isolated in the first half of 2017 were all serotype 8b, which was related to the widespread application of inactivated serotype 4 adenovirus vaccines. These results demonstrate that fowl adenovirus serotypes 11, 4, and 8b were the predominant serotypes in some regions of China between 2007 and June 2014, between June 2014 and 2016, and in the first half of 2017, respectively. Layers were the predominant host infected with fowl adenovirus serotype 4 and could also be infected by serotype 11.

The bactericidal efficacy of food additive–grade calcium hydroxide [FdCa(OH)₂] was evaluated for inactivation of Salmonella Infantis and Salmonella Enteritidis in liquid and Salmonella Infantis on contaminated eggshells. The activity of FdCa(OH)₂ was also compared with that of sodium hypochlorite (NaOCl) containing 150 ppm chlorine (150 ppm NaOCl). FdCa(OH)₂ solutions (0.1% and 0.2%) in the presence or absence of organic materials (5% calf serum [CS]) at pH 12.6 were used to inactivate Salmonella Infantis and Salmonella Enteritidis in a reaction tube or on eggshells artificially contaminated with Salmonella Infantis. Both 0.1% and 0.2% FdCa(OH)₂ were capable of inactivating Salmonella Infantis and Salmonella Enteritidis in liquid at >3 log₁₀ colony-forming units (CFU)/ml within 3 and 1 min of contact time, respectively, even in the presence of 5% CS. Additionally, 0.1% and 0.2% FdCa(OH)₂ reduced bacterial levels on contaminated eggshells to >3 log₁₀ CFU/ml, within 3 and 1 min, respectively, in the presence of 5% CS. Without CS, 0.1% and 0.2% FdCa(OH)₂ could reduce bacteria on eggshells to >3 log₁₀ CFU/ml within 1 min and 30 sec, respectively. In contrast, 150 ppm NaOCl solution could not inactivate bacteria on eggshells down to >3 log₁₀ CFU/ml within 3 min contact time, either with or without CS, and no bacterial reduction was observed in redistilled water. The findings of the present study indicate that FdCa(OH)₂ solution has high efficacy against foodborne bacteria and may be a good candidate for enhancement of biosecurity at farms and egg processing plants.

Since being successfully reintroduced into Ontario, Canada, wild turkey (Meleagris gallopavo) populations have undergone robust growth and range expansion. This, along with increases in land use changes from human population growth and subsequent developments in agriculture and livestock production, has heightened opportunities for interactions between wild turkeys, domestic poultry, and humans. As conspecifics, wild and domestic turkeys are susceptible to infection and disease from many of the same pathogens. Thus, transmission by direct or indirect contact is a potential health threat to both groups, particularly with the overlapping range of wild turkeys in Ontario with numerous commercial and backyard poultry operations. However, these threats are difficult to assess due to knowledge gaps in the prevalence and geographic distribution of potential pathogens circulating among wild turkeys. We assessed for potentially pathogenic bacteria in free-ranging, hunter-harvested wild turkeys in Ontario (n = 152) by cloacal swab culture for Campylobacter spp., Salmonella spp., and Escherichia coli and culture of lung and spleen for Pasteurella multocida, Ornithobacterium rhinotracheale, and Erysipelothrix rhusiopathiae. Antimicrobial resistance testing was also performed on E. coli isolates. Generic E. coli isolates were recovered from 69.1% (105/152) of wild turkeys tested, and two (1.9%) of these isolates exhibited resistance to azithromycin and one (1.0%) to ampicillin. Intermediate susceptibility to chloramphenicol was observed in one (1.0%) isolate. One (0.7%) wild turkey swab tested positive for C. jejuni, but no samples were positive for P. multocida, Salmonella spp., O. rhinotracheale, or E. rhusiopathiae. To our knowledge, this is the first survey of these bacteria and assessment for antimicrobial resistance among wild turkeys in Ontario.

Preliminary diagnosis of clinical symptoms and gross lesions with subsequent histopathologic and PCR analyses revealed histomoniasis in 276 chicken flocks in Jiangsu Province, China, and surrounding areas from January 2012 to December 2015. Detailed statistical analysis was performed to explore the occurrence and epidemic characteristics of histomoniasis in chicken flocks. The results indicated that histomoniasis usually occurred in free-range flocks of local broilers and laying hens. Also, 2- to 3-mo-old chickens were most susceptible to infection, and adult chickens rarely developed infection. The morbidity rate of chickens was generally 10%–30%, with mortality rates of less than 10%. Moreover, histomoniasis is a seasonal disease, occurring most often from April to June, and the rate of coinfection with heterakids in the ceca of infected chicken was 50.8%. The symptoms of diseased chickens included mental fatigue, bowing of the head and wings, and yellowish green droppings, with bloody stool in very limited cases. Most of the pathologic changes were characteristic of the disease, but there were also some atypical lesions confirmed by laboratory techniques. In the current study, the histomoniasis epidemic was first investigated in Chinese chicken flocks, and the results provided a useful reference for further study of this disease.

To assess the survival of Gallibacterium anatis in dead laying hens, 21-wk-old laying hens were injected intraperitoneally with 0.5 ml brain hearth infusion broth containing 10⁸ colony-forming units (CFU) of G. anatis 12656-12 liver (n = 16), Escherichia coli ST141 (n = 16), or a mix of G. anatis 12656-12 liver and E. coli ST141 (n = 16), respectively. Birds were euthanatized 24 hr post injection. From each group eight dead birds were kept at 4 C and eight at room temperature. Swab samples were taken at different time points post euthanatization and streaked on blood agar plates. From the birds kept at 4 C, G. anatis was reisolated from the G. anatis and the G. anatis–E. coli co-injected groups at least 12 days post euthanization. From birds kept at room temperature, G. anatis was reisolated up to 2 days post euthanatization. When using the gyrB-based G. anatis–specific quantitative PCR (qPCR), G. anatis was detected within at least 5 days, and up to 5 days post euthanatization, from birds kept at room temperature and 4 C, respectively. Escherichia coli was reisolated from all the time points independent of how the birds were kept. No difference was observed between the reisolation rates for G. anatis or E. coli when comparing similar detection methods. For birds kept at 4 C, bacterial cultivation was a more sensitive method for detecting G. anatis (P < 0.05), whereas for birds kept at room temperature, the G. anatis–specific qPCR outperformed bacterial culture (P < 0.05). In conclusion, we demonstrated that G. anatis has a poorer survival rate than does E. coli in dead chickens kept at room temperature. That finding may affect the overall diagnostic sensitivity and lead to underdiagnosis of G. anatis in a normal production setting.

An expert elicitation was staged to rapidly decipher plausible routes and risks of pathogen transmission in the 2017 H7N9 avian influenza (AI) outbreak in the four-state region of Tennessee, Alabama, Georgia, and Kentucky. The process included the identification of risk factors found in a preponderance of commercial broiler breeder case farms over matched controls and an opinion-based weighting of risks and mitigations perceived influential to this outbreak. Although the two highly pathogenic AI case farms had general location and company ownership in common, obvious connections were lacking for the remainder of H7N9-infected (all low pathogenicity) commercial farms. Expert elicitation of differences between known cases and controls suggested a key role for environmental rather than lateral (business network) pathways in the distribution of low pathogenicity AI across commercial broiler breeder operations. Factors with greatest strength as predictors of disease, whether or not they were causal, included mesopredator or rodent incursions, enclosure defects, and habitat disturbance that might attract wildlife to the farm (e.g., feed spills and vacating of neighboring properties). Business affiliations that may have facilitated farm-to-farm transfer, in contrast, were limited. Biosecurity standards varied across this study group but were no more or less stringent among cases over controls. However, results from a parallel hypothetical scenario staged to address field data gaps suggest that uniformity and consistency in the implementation of biosecurity practices may impact risk of disease introduction. Importantly, this study was conducted within a few weeks and with little disruption to emergency response activities. As such, the approach offers an alternative model for interim field investigation of new or emerging high-consequence diseases with immediate decision support needs.

We identified novel linear epitopes on the infectious bronchitis virus (IBV) spike S2 region. The conformational structure of the IBV spike protein was predicted from a homologous protein, human coronavirus NL63 spike. Although the obtained structure was incomplete, most of the IBV spike protein structure was predicted; the N-terminus of the S1 region could not be predicted due to its variability. In the model, the region located in the proximity of the fusion peptide appeared to be well conserved, and we evaluated the antigenicity of these domains, which are involved in the membrane fusion machinery. Western blotting revealed that IBV TM86 spike residues 686–723 were antigenic. Epitope mapping analysis using synthesized peptides revealed that IBV TM86 spike 669–685 (SNFSTGAFNISLLLTPP), 686–697 (SNPRGRSFIEDL), and 692–703 (SFIEDLLFTSVE) residues were major linear epitopes; two identified epitopes (686–697 and 692–703) were covered by the fusion peptide, and the other epitope (669–685) was adjacent to the fusion peptide. Although the identified epitopes are identically located as the neutralizing epitope in severe acute respiratory syndrome coronavirus, the recombinant protein that includes those epitopes could not elicit neutralizing antibodies against IBV. This is the first report describing IBV spike S2 epitopes located in the proximity of the fusion peptide, and it is suggested that the spike fusion machinery of IBV may differ from that of severe acute respiratory syndrome coronavirus, or, alternatively, IBV may have another mechanism to penetrate the cell membrane.

In recent years, acute severe outbreaks of infectious bursal disease (IBD) are frequently observed in commercial chicken populations of the North East Region (NER) of India, resulting in huge economic loses to poultry farmers. Field outbreaks of IBD in 30 different poultry farms in the NER were confirmed by clinicopathologic examination and reverse transcriptase PCR. A total of 10 isolates of IBD virus (IBDV) from these outbreaks were characterized by the genetic analysis of VP1 and the hypervariable region of the VP2 gene. Nucleotide sequences, deduced amino acid sequences, and phylogenetic analysis of both VP2 and VP1 genes revealed two genetically diverse strains of very virulent IBDV (vvIBDV) and one intermediate strain circulating in the NER. These isolates differ at nucleotide and amino acid levels from vvIBDV isolates of mainland India and are clustered in distinctly separate groups in the phylogenetic tree. Six of the isolates revealed a unique combination of vvIBDV amino acid signatures in the VP2 gene (A222, I256, I294), while bearing the non-vvIBDV amino acid signatures of the VP1 gene (146E, 147G, 242D), but they are clearly classified as vvIBDV in a phylogenetic analysis of both genes. Interestingly, one of the isolates showed 99% sequence homology with attenuated vaccine strains in the VP2 gene and clustered together. This study demonstrates the diversity of IBDVs in India and document for the first time the possible involvement of attenuated vaccine strains in the epidemiology of IBD in India.

Although infectious bronchitis virus (IBV) has been described as one of the most economically important viral respiratory diseases in poultry, there are few analyses of outbreaks that use spatial statistics. In order to better understand how the different genotypes of IBV behave spatially and temporally, we used geographic information system-based mapping coupled with spatial and spatial-temporal statistics to identify statistically significant clustering of multiple strains of infectious bronchitis (IB) between 2008 and 2012 in California. Specifically, space-time permutation and multinomial models were used to identify spatial and spatial-temporal clusters of various genotypes of IBV. Using time permutations (i.e., windows) spanning days to years, we identified three statistically significant (P < 0.05) clusters. In contrast, multinomial models identified two statistically significant spatial-temporal clusters and one statistically significant spatial cluster. When comparing the space-time permutation and multinomial models against each other, we identified spatial and temporal overlap in two of the three statistically significant clusters. From a practical perspective, multinomial clustering approaches may be advantageous for studying IB because the model allows the different genotypes of IB to be independent nominal variables, thereby allowing for a more detailed spatial analysis. To that point, based on their risk ratios, the genotypes classified as vaccine-related were identified as the most significant contributor to two of the three mutinomial clusters. Additionally, statistically significant clusters were mapped and layered on a hot-spot analysis of commercial poultry farm density in order to qualitatively assess the relationship between farm density and clusters of IBV. Results showed that one of the three space-time permutations and one of the three multinomial clusters were spatially centered near the highest density farm areas, as determined by the hot-spot analysis.

To evaluate the virulence of avian pathogenic Escherichia coli (APEC) isolates obtained from colibacillosis cases associated with pericarditis, perihepatitis, and salpingitis, the embryo lethality assay and experimental infection model in chicks were used in this study. According to the established criteria based on mortality in the embryo lethality assay for evaluating the virulence of E. coli isolates, 23 of the 26 APEC isolates associated with pericarditis and perihepatitis and 8 of the 20 isolates associated with salpingitis were found to be virulent. Isolate D137, which had been obtained from a case with pericarditis and perihepatitis and had an embryo mortality of 92%, and isolate D445, which had been obtained from a case with pericarditis and perihepatitis and had an embryo mortality of 17%, were used for the experimental infection. Four of the five 11-day-old chickens inoculated through the air sac with isolate D137 died 1 day postinoculation, and the challenge strain was recovered from the air sac, pericardial sac, or liver; however, colibacillosis lesions were found in only one of the five birds postmortem. All five chicks inoculated with isolate D445 survived for 7 days postinoculation and exhibited airsacculitis or pericarditis lesions at 7 days postinoculation; the challenge strain was not recovered from the lesions postmortem. The results obtained in this study suggest that the different APEC isolates tested cause illness in chickens through distinct pathogenesis.

The connectedness in Arctic regions between migratory waterbird populations originating from different continents and the potential for virus exchange at their shared Arctic breeding ground point to the need to explore the largely unstudied circumpolar circulation of avian influenza viruses (AIV). We here report the investigation of AIV in wild birds and lakes in a high Arctic area of Northeast Greenland. No AIV could be detected in the fecal, feather, and water samples collected from large flocks of pink-footed geese Anser brachyrhynchus and barnacle geese Branta leucopsis in and around refuge lakes, where they congregate at high density during their flightless molting period in summer.

Focal duodenal necrosis (FDN) is an intestinal disease of egg-laying chickens, characterized by multifocal mucosal erosions in the duodenal loop and proximal jejunum. It is currently considered by the Association of Veterinarians in Egg Production and the United States Animal Health Association as one of the top five disease concerns of the table egg industry in the United States. Previous studies have associated this condition with Clostridium species. The purpose of this study was to investigate the epidemiologic characteristics of table egg layer flocks affected with FDN. An online questionnaire was distributed to commercial layer operations in different states in the United States. Layer farms that had diagnosed FDN within the past 12 mo were surveyed. The questionnaire had 45 questions about management, nutrition, housing, and methods for disease prevention and control. Thirty-seven surveys were sent and 21 were completed, which represents a response rate of 56.7%. The survey results showed the presence of FDN in five egg-layer genetic lines or breed crosses of different ages, with most cases reported between 30–39 wk of age. The pullets were cage-reared in all affected flocks and the majority of flocks in production were housed in traditional cages. Most of the FDN-affected flocks received more than 12 different feed formulations from pre-lay to 60 wk of age. Distiller's dried grain with solubles was a common ingredient added to the feed in the majority of affected flocks, and all flocks were provided with limestone as a calcium source for egg production. Most surveys reported that coccidiosis and roundworm parasitism were not problems in affected flocks in production; however, pests such as flies and rodents were reported as problems in most affected flocks. Additionally, most affected farms never washed feeders, cages, and houses before disinfection, which may not be sufficient to prevent the persistency and transmission of the causative agent of FDN. In conclusion, several management practices that have been associated with enteric disease, including clostridial-associated enteritis, were described by the majority of FDN-affected flocks. Additional studies are needed to determine if management and health practices identified in this survey represent risk factors for FDN.

Sixty-four cases of white chick syndrome (WCS) in broiler breeders producing affected progeny were reported from seven hatcheries in Ontario, Canada, between 2009 and 2016, with 43 of those originating from two hatcheries owned by a single company. WCS cases were identified by the presence of typical chicks in the hatchery that were generally weak with pale to white down, enlarged abdomens, and occasionally brown wiry fluff on the dorsum of the neck. Affected embryos and chicks had characteristic gross and histologic liver lesions, and livers were positive for chicken astrovirus (CAstV) RNA by real-time reverse transcriptase PCR. Affected broiler breeder flocks experienced egg production drops of 0% to 21% and hatchability drops of 0% to 68.4%. The amino acid sequence of the region encoding the capsid gene of WCS viruses demonstrated all Ontario CAstV to be in Group B, Subgroup Bii.