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Viruses within the Coronaviridae family show variations within their genome sequences, especially within the major structural protein, the Spike (S) glycoprotein gene. Therefore, many different antigenic forms, serotypes, or variant strains of avian coronaviruses (AvCoV) exist worldwide. Only a few of them, the so called protectotypes, cross protect against different serotypes. New serotypes arise by recombination or spontaneous mutations. From time to time, antigenic virus variants appear which differ significantly from known serotypes. The result of this variability is an inconsistent nomenclature and classification of virus strains. Furthermore, there are currently no standard classification methods defined.

Within the framework of the COST Action FA1207 “Towards control of avian coronaviruses: strategies for diagnosis, surveillance, and vaccination” (working groups “Molecular virology” and “Epidemiology”), we aimed at defining and developing a unified and internationally standardized nomenclature and classification of AvCoVs. We recommend the use of “CoV Genus/AvCov/host/country/specimen id/year” to refer to AvCoV strains.

Outbreaks of H5 highly pathogenic avian influenza (HPAI) in commercial poultry are a constant threat to animal health and food supplies. While vaccination can enhance protection and reduce the spread of disease, there is considerable evidence that the level of immunity required for protection varies by subtype and virulence of field virus. In this study, the efficacy of a recombinant turkey herpesvirus (rHVT) vector vaccine expressing the hemagglutinin gene from a clade 2.2 AI virus (A/Swan/Hungary/4999/2006) was evaluated in turkeys for protection against challenge with A/Whooper Swan/Mongolia/L244/2005 H5N1 HPAI clade 2.2. One-day-old turkeys received a single vaccination and were challenged at 4 wk postvaccination with 2 × 10⁶ 50% embryo infectious dose per bird. The results demonstrate that following H5N1 HPAI challenge 96% protection was observed in rHVT-AI vaccinated turkeys. The oral and cloacal swabs taken from challenged birds demonstrated that vaccinated birds had lower incidence and titers of viral shedding compared with sham-vaccinated birds. From respiratory and gastrointestinal tracts, there was a greater than 6 log₁₀ reduction in shedding in vaccinated birds as compared with the controls. This study provides support for the use of a commercially available rHVT-AI vaccine to protect turkeys against H5N1 HPAI.

We previously demonstrated that adaptation of an embryo-attenuated infectious bronchitis virus (IBV) Arkansas Delmarva Poultry Industry (ArkDPI)–derived vaccine to chicken embryo kidney (CEK) cells shifted the virus population towards homogeneity in spike (S) and nonstructural protein genes. Moreover, the typical Ark vaccine subpopulations emerging in chickens vaccinated with commercial Ark vaccines were not detected in chickens vaccinated with the CEK-adapted virus. In this study, chickens vaccinated with a low dose (1.6 × 10³ EID₅₀/bird, where EID₅₀ is 50% embryo infectious dose) of CEK-adapted Ark vaccine at 5 days of age showed a significant reduction of IBV RNA in lachrymal fluids and decreased incidence of IBV RNA detection in tracheal swabs 5 days after challenge compared to unvaccinated challenged chickens. In a second experiment, 5-day-old chickens were vaccinated with 10⁴ or 10⁵ EID₅₀/chicken of CEK-adapted Ark vaccine, and protection was compared to chickens vaccinated with 10⁵ EID₅₀/chicken of the commercial ArkDPI-derived vaccine from which the CEK-adapted virus originated. All vaccinated chicken groups showed a significant reduction of respiratory signs and viral load 5 days after Ark virulent challenge compared to unvaccinated challenged controls. No viral subpopulations different from the challenge virus were detected in chickens vaccinated with CEK-Ark after challenge. In contrast, IBV S1 sequences differing from the predominant population in the challenge virus were detected in several chickens vaccinated with the commercial Ark attenuated vaccine. From an applied perspective, the CEK-adapted IBV ArkDPI-derived vaccine is an improved and effective vaccine candidate with which to protect chickens against virulent Ark-type strains.

On the basis of the data from the California Animal Health and Food Safety Laboratory System, 1444 infectious bronchitis (IB) cases were diagnosed between 1997 and 2012. Epidemiologic analyses demonstrated two major IB virus (IBV) outbreak peaks, affecting mainly 35-to-49-day-old broiler chickens. California variant 1737 (CA1737) and California variant 1999 (Cal 99) IBV types were the most prevalent genotypes during the analyzed period. To further understand the increased prevalence of these genotypes, we assessed and compared the variability of the S1 gene hypervariable region of CA1737 and Cal 99 with the variability of IBV strains belonging to the Massachusetts 41 (M41) and Arkansas (Ark) types during serial passages in embryonated chicken eggs. On the basis of the S1 nonsynonymous changes, seven different subpopulations were detected in M41. However, the predominant population of the field strain M41 before passages continued to be predominant throughout the experiment. In contrast, Ark passaging resulted in the detection of 13 different subpopulations, and the field sequence became extinct after the first passage. In IBV Cal 99, eight different subpopulations were detected; one of these became predominant after the second passage. In CA1737, 10 different subpopulations were detected. The field strain major sequence was not detected after the first passage but reappeared after the second passage and remained at low levels throughout the experiment. Compared with M41 and Ark, Cal 99 and CA1737 showed intermediate variability.

The majority of bacterial diarrhea-causing illnesses in domestic pigs result from infection with Escherichia coli, Salmonella spp., or Campylobacter spp. These bacterial enteropathogens also correspond with the most-common bacteria isolated from wild birds. Additionally, viral pathogens such as avian influenza virus (AIV), West Nile virus (WNV, including Kunjin disease), and Newcastle disease virus (NDV) may also be carried and transmitted by birds in Australia. Introduced European starlings (Sturnus vulgarus) are one of the most-frequently reported birds on piggeries in Australia. The presence of the three bacterial pathogens, Salmonella spp., Campylobacter spp., and Escherichia coli, as well as the three viral pathogens AIV, WNV, and NDV, were evaluated in starlings captured on four commercial piggeries in South Australia. A total of 473 starlings were captured on the four piggeries in 2008 and 2009. A cloacal swab was taken from each bird and cultured for bacterial identification, with follow-up serotyping of any positives, whilst fifty samples were analyzed by PCR for the three target viral pathogens. There was no AIV, WNV, or NDV detected in the 50 starlings sampled. Escherichia coli was found to be present in the starling populations on all four piggeries whilst Salmonella spp. and Campylobacter jejuni were found to be present only in the starling population sampled on one piggery. Serotyping identified pig-pathogenic strains of the bacteria. The prevalence of these production-limiting bacterial pathogens in starlings, coupled with the large starling populations often found inside piggeries during daylight hours in the summer months, presents a disease transmission risk and jeopardizes piggery disease management. Removal of starlings from agricultural enterprises (as shown by international studies), or prevention of starling access to animal feed and water, could substantially reduce the risk of transmission of enterobacterial pathogens from starlings to livestock.

In April 2014, poor fertility in a major commercial goose breeder operation in California triggered the submission of six live affected Toulouse ganders (Anser anser) to the California Animal Health and Food Safety Laboratory, Turlock branch (University of California–Davis). Toulouse were principally affected among all breeds, and their egg fertility dropped from 65.7% to less than 33.9% in the first 40 days of the 2014 breeding season. The flock consisted of 410 adult birds, 90 males and 320 females, between 2 and 5 yr of age. Inspection of the flock revealed that 44.4% of the Toulouse ganders had severe phallic deformities that prevented them from mating. At postmortem examination, severe yellowish fibrocaseous exudate disrupted the architecture of the phallus and occasionally produced fistulating tracts through the wall of the organ. Microscopically, multifocal lymphoid nodules were noted in the mucosa and submucosa of the phallus and were associated with extensive granulomatous reaction, intralesional bacteria, and spermatozoa. Mycoplasma spp. were isolated from the phallus of affected and nonaffected birds, and PCR protocols targeting the 16S–23S ribosomal RNA intergenic spacer regions and the RNA polymerase beta subunit gene were performed to identify the isolates. Three distinct species were identified on sequencing and analysis using the National Center for Biotechnology Information basic local alignment search tool: Mycoplasma cloacale, Mycoplasma anseris, and an unknown novel Mycoplasma sp. Additionally, Pasteurella multocida, in combination with other bacteria, was also isolated from the phallic lesions and identified as serotype 3 with a DNA profile of 1511 (National Veterinary Service Laboratory). This is the first report of these Mycoplasma spp. and other bacteria associated with reproductive disease in ganders in the United States.

Clostridium perfringens infection causes subclinical and clinical necrotic enteritis in poultry flocks, and it is estimated to result in US$2 billion of losses worldwide every year. The aims of this study were to determine the incidence, toxin types, and antimicrobial resistance levels to C. perfringens isolated from premarket, 5-wk-old, clinically healthy broiler chickens in Taiwan, and to examine the relationships between intestinal lesions and the numbers of C. perfringens in intestinal contents. In total, 435 samples of chicken ileum contents were collected from 98 broiler farms during June 2012 to February 2013. The C. perfringens isolation rate was 9.9% (43/435). The positive rate of tested farms was 29.6% (29/98). All the isolates were C. perfringens type A, only possessing the cpa gene encoding for toxin α. No netB gene encoding NetB toxin associated with necrotic enteritis, and no cpe gene encoding for the C. perfringens enterotoxin causing human intestinal disorder were detected. A quantitative PCR analysis revealed that the mean C. perfringens number in the intestinal contents was 3.9 × 10⁶ colony-forming units (CFU)/g, ranging from 6.85 × 10² to 1.61 × 10⁷ CFU/g. The gross and histopathologic lesions revealed a positive correlation (p < 0.05) between lesion score and C. perfringens number in the ilea of C. perfringens–positive chickens. Antimicrobial susceptibility tests of all C. perfringens isolates indicated that the minimum inhibitory concentration inhibiting 50% of isolates (MIC₅₀) for amoxicillin, bacitracin, chlortetracycline, enrofloxacin, erythromycin, florfenicol, and lincomycin was ≤0.125, 0.5, 128, 0.25, ≥256, 2, and ≥256 μg/ml, respectively. Most of the C. perfringens isolates were susceptible to amoxicillin, bacitracin, and enrofloxacin but resistant to chlortetracycline, erythromycin, and lincomycin. Interestingly, C. perfringens isolated from chickens with severe lesions had higher MIC₅₀ for erythromycin and lincomycin than those isolates from chickens with mild lesions. Conclusively, reductions in both the incidence of C. perfringens infection on farms and the concentrations of C. perfringens in intestines to improve broiler health are still needed in Taiwan.

SUMMARY. We showed here that an H5N8-subtype highly pathogenic avian influenza virus (HPAIV) was transmitted to both the internal contents and shells of eggs laid by white leghorn hens experimentally infected with the virus. Seven of eight HPAIV-infected hens laid eggs until 4 days postinoculation (dpi). The mean number of eggs laid per head daily decreased significantly from 0.58 before inoculation to 0.18 after viral inoculation. The virus was detected in the eggs laid by three of the seven hens. Viral transmission was detectable beginning on 3 dpi, and virus titers in tracheal and cloacal swabs from the hens that laid the contaminated eggs exceeded 2.9 log₁₀ EID₅₀. The level of viral replication and its timing when virus replicates enough to be detected in oviduct after virus inoculation appear to be key factors in the transmission of H5N8 HPAIV from infected hens to laid eggs.

Salmonella and the poultry red mite (Dermanyssus gallinae) remain very challenging diseases for the poultry industry worldwide because of the inefficiency of implementing and integrating eradication and control programs, which results in very high economic losses to the poultry industry. The aim of this study was to determine the association between biosecurity levels in layer farms and the occurrence of both D. gallinae and Salmonella spp., as well as the relationship between D. gallinae infestations on farms and Salmonella occurrence. For this purpose, 22 layer farms using the common battery cage housing system in different parts of Kosovo were randomly selected and analyzed for the presence of D. gallinae and Salmonella in samples, such as feces, water, feed, and dust. Two pooled samples of D. gallinae (2n = 100) were directly analyzed for the presence of Salmonella in the outer and inner parts of cuticula from D. gallinae. A chi-square test was used to determine the association between experience in poultry production, rearing, and level of different biosecurity elements in relation to the occurrence of D. gallinae and Salmonella. Dermanyssus gallinae was found on 15 farms, whereas Salmonella was found on eight of those 15 farms from different environmental samples and on one farm where D. gallinae was not found. In two pooled samples Salmonella was isolated directly from the inner part of the cuticula from D. gallinae, which represents the first direct isolation of Salmonella from D. gallinae mites. Association between the level of biosecurity and the occurrence of D. gallinae and Salmonella was strong. The study indicates that proper biosecurity measures should be in place to lower the occurrence of D. gallinae and Salmonella.

A case-control study was conducted among commercial table-egg layer and pullet operations in Iowa and Nebraska, United States, to investigate potential risk factors for infection with highly pathogenic avian influenza (HPAI) H5N2. A questionnaire was developed and administered to 28 case farms and 31 control farms. Data were collected at the farm and barn levels, enabling two separate analyses to be performed—the first a farm-level comparison of case farms vs. control farms, and the second a barn-level comparison between case barns on case farms and control barns on control farms. Multivariable logistic regression models were fit using a forward-selection procedure. Key risk factors identified were farm location in an existing control zone, rendering and garbage trucks coming near barns, dead-bird disposal located near barns, and visits by a company service person. Variables associated with a decreased risk of infection included visitors changing clothing, cleaning and disinfecting a hard-surface barn entryway, and ceiling/eaves ventilation in barns.

Between December 2014 and June 2015, an outbreak of H5N2 HPAI caused the largest and most expensive agriculture emergency in U.S. Department of Agriculture–Animal and Plant Health Inspection Service history. The outbreak affected 21 states; 232 poultry farms (211 commercial and 21 backyard) were affected, and approximately 49.6 million birds were depopulated on poultry farms. The majority of affected farms were commercial turkey operations (n = 160). This report is a case series describing 104 H5N2 HPAI-affected turkey farms in Iowa, Minnesota, Missouri, North Dakota, South Dakota, and Wisconsin that had H5N2 HPAI virus detected between March 5 and June 1, 2015. The farm manager or farm personnel voluntarily completed an epidemiologic questionnaire administered by state and federal animal health officials. Equipment and vehicle sharing with other farms was common, particularly for feed trucks (77% of farms shared feed trucks with other farms), live haul loaders (90.4%), poult trailers (72.0%), and preloaders (80.7%). Many farms had water bodies in proximity to the farm, such as a pond (42.6%) or stream (21.8%). About one-third of farms (33.7%) reported seeing wild birds inside the turkey barns. Only 44.2% of farms reported that third-party biosecurity audits or assessments had been conducted. Because the newly introduced Asian H5N8 HPAI and two new HPAI viruses, H5N2 and H5N1, are now circulating in U.S. wild birds, primarily migratory waterfowl, a greater potential for reoccurrence exists with the spring and fall migratory seasons, representing higher risk periods for outbreaks of HPAI in commercial poultry farms in the future. Eliminating exposure to wild birds, especially waterfowl or environments contaminated by wild waterfowl, will reduce risk of reintroduction of H5N2 HPAI virus, and ensuring good on-farm biosecurity will help the poultry industry avoid introduction of influenza and lateral spread between farms.

Herpesvirus of turkeys (HVT) is a widely used vector for poultry vaccines. However, different HVTs expressing different foreign antigens cannot always be used simultaneously because of the risk of recombination and interference. In this study, we inoculated a mixture of an HVT-expressing the antigen of Newcastle disease virus (NDV; HVT/ND) and Marek's disease virus (MDV) serotype 1 Rispens virus expressing the antigen of infectious bursal disease virus (IBD; Ripens/IBD) into chickens. This mixture showed 94%, 100%, or 94% protection against MDV, IBDV, or NDV challenge, respectively. In conclusion, the combination of Rispens/IBD and HVT/ND is effective for vaccination against MDV, IBDV, and NDV without significant interference.

Biosecurity measures are the first line of defense against highly pathogenic avian influenza (HPAI) on farms. It is generally recognized that an individual's behavior can be influenced by the knowledge they possess. However, empirical study has not reported an association between poultry producers' awareness of HPAI symptoms and their actual biosecurity actions. The aim of this study is to classify knowledge items of HPAI by exploratory factor analysis (EFA) and to examine the determinants of different types of knowledge and the effect of different types of knowledge on biosecurity preventive behaviors (BPBs). The survey (n = 297) was conducted using a questionnaire to measure the level of awareness of items related to HPAI and the actual adoption of BPBs among poultry farmers in the Chinese province of Jiangsu. The EFA revealed three main types of knowledge, which were categorized as avian influenza (AI) epidemic characteristics, primary biosecurity preventive knowledge (basic biosecurity preventive knowledge against AI), and essential biosecurity preventive knowledge (crucial biosecurity preventive knowledge against infection of AI). Multivariate regression showed that only poultry farmers' awareness of essential biosecurity preventive knowledge was positively associated with their actual BPBs. Additionally, educational attainment, number of years of experience raising poultry, farming operation size, and training were associated both with BPB and most of the knowledge factors or knowledge items. Training of existing poultry farmers is probably a feasible scheme; furthermore, the training should focus on the essential biosecurity preventive knowledge. On the other hand, policy initiatives to encourage large-scale poultry farming while discouraging small-scale backyard poultry husbandry would be an effective method of improving the management standards of rural poultry farming.

Airborne pathogens can cause infections within parrot (Psittaciformes) and pigeon (Columbiformes) holdings and, in the case of zoonoses, can even spread to humans. Air sampling is a useful, noninvasive method which can enhance the common sampling methods for detection of microorganisms in bird flocks. In this study, fecal and air samples were taken from four parrot holdings. Additionally, cloacal and oropharyngeal swabs as well as air samples were taken from 15 racing pigeon holdings. Parrots were examined for psittacine beak and feather disease virus (PBFDV), proventricular dilatation disease virus (PDDV), adenoviruses (AdVs), avian paramyxovirus type-1 (APMV-1), avian influenza virus (AIV), Chlamydia psittaci (CP), and Mycobacterium avium complex (MAC). MAC and AdVs were detected in three parrot holdings, CP was detected in two parrot holdings, and PBFDV and PDDV were each detected in one parrot holding. Pigeons were examined for the pigeon circovirus (PiCV), AdVs, and CP; PiCV and AdVs were detected in all investigated pigeon holdings and CP was detected in five pigeon holdings.

The H9N2 subtype of low pathogenic avian influenza (LPAI) virus is the most prevalent LPAI in domestic poultry. We previously reported the natural reassortant H9N2 viruses between North American and Eurasian lineages isolated from wild birds in Korea. These viruses were identified in China and Alaska, providing evidence of intercontinental dispersal. In this study, we evaluated the infectivity, transmissibility, and pathogenic potential of these H9N2 viruses and Eurasian H9N2 virus identified from wild birds using specific-pathogen-free chickens. Three-week-old chickens were infected intranasally. All of these reassortant H9N2 viruses could not be replicated and transmitted in chickens. On the other hand, three out of eight chickens inoculated with the Eurasian H9N2 virus shed detectable levels of virus and showed seroconversion but did not show contact transmission of the virus. Although all reassortant H9N2 viruses could not be replicated and transmitted in chickens, and although there are no reports on reassortant H9N2 virus infection in poultry farms until now, monitoring of reassortant H9N2 viruses should be continued to prepare for the advent and evolution of these viruses.

Serologic tests are a valuable tool for retrospective surveillance of avian influenza viruses (AIV) and monitoring of postvaccination host immune response. Yet collection of serum samples, particularly in adult breeder chickens, is laborious, intrusive to birds, and may pose a serious risk to the biosecurity of a flock. In this study we compared the level of AIV-specific antibody titers in eggs and serum samples obtained from broiler breeder chickens vaccinated at 6, 12, and 18 wk of age with H5N2-inactivated vaccine. Nucleocapsid protein-specific ELISA and hemagglutination inhibition test (HI) against homologous as well as heterologous antigens were used. The eggs and sera were collected at 22, 30, 45, and 50 wk of age (i.e., 4, 12, 27, and 32 wk after the third and final immunization, respectively). Using ELISA, the number of positive egg yolk samples decreased over time after vaccination, from 97% to 47%, while the seropositivity rate of serum samples was 97%–100% during the whole investigation period. No antibody titers were detected in egg white. By HI, antibody titers in serum samples were higher than in egg yolk samples. Compared to the homologous H5N2 antigen, significantly lower HI titers were obtained by using a heterologous H5N1 virus of clade 2.2.1.2. In addition, no HI titers were detected in egg yolk and/or serum samples tested against the antigen of an Egyptian H5N1 antigenic drift variant of clade 2.2.1.1. This study indicates that egg yolk may be used to monitor the postvaccination immune status of broiler breeder chickens and retrospective serosurveillance—by HI when a matching antigen is available as well as by ELISA—particularly for up to 12 wk postvaccination.

Recent metagenomic analyses of the enteric viromes in turkeys and chickens have revealed complex viral communities comprised of multiple viral families. Of particular significance are the novel avian picobirnaviruses (family Picobirnaviridae), multiple genera of tailed phages (family Siphoviridae), and undescribed avian enteric picornaviruses (family Picornaviridae). In addition to these largely undescribed—and therefore relatively poorly understood—poultry enteric viral families, these metagenomic analyses have also revealed the presence of well-known groups of enteric viruses such as the chicken and turkey astroviruses (family Astroviridae) and the avian rotaviruses and reoviruses (family Reoviridae). The order Picornavirales is a group of viruses in flux, particularly among the avian picornaviruses, since several new genera have been described recently based upon community analysis of enteric viromes from poultry and other avian species worldwide. Our previous investigation of the turkey enteric picornaviruses suggests the avian enteric picornaviruses may contribute to the enteric disease syndromes and performance problems often observed in turkeys in the Southeastern United States. This report describes our recent phylogenetic analysis of turkey and chicken enteric samples archived at the Southeast Poultry Research Laboratory from 2004 to present and is a first step in placing these novel avian picornaviruses within the larger Picornaviridae family.

Very virulent infectious bursal disease virus (vvIBDV) was diagnosed in a pullet farm in Washington in 2014. Infectious bursal disease virus is resistant to many environmental stresses and often persists on farms for months. There have been conflicting reports as to whether composting can destroy vvIBDV in the manure. This project investigated the composting of litter from the affected house using an aerated static pile to inactivate the virus. Two weeks before the affected pullet flocks were moved to the layer house, specific-pathogen-free (SPF) birds were placed in the barns. Ten days after they were placed, three SPF birds died and were positive for vvIBDV. Thirty percent of the SPF birds were positive for vvIBDV. After the pullets were moved, at 20 wk of age, the litter in the house was composted using the aerated static pile method. The pile was maintained at above 55 C for 4 wk. After this time, 30 additional SPF birds were placed on the composted material. Two weeks later, the birds were healthy and there was no evidence of vvIBDV. The subsequent pullet flock did not break with vvIBDV. These results demonstrate that this composting method can be used to decontaminate the litter from vvIBDV and help prevent the spread of vvIBDV.

Avian encephalomyelitis (AE) was diagnosed in three flocks of leghorn layer pullets following AE vaccination. Ages of the birds were 11, 12, and 14 wk. The submissions came from three different companies located in two geographic areas of the Central Valley of California. The clinical signs included birds down on their legs, unilateral recumbency or sitting on their hocks, lethargy, reluctance to move, dehydration, unevenness in size, low weight, tremors of the head in a few birds, and mildly to moderately elevated mortality. The flocks had been vaccinated against fowl pox and AE with a combined product in the wing-web 2 wk prior to the onset of AE clinical signs. Histopathologic examination revealed lesions consistent with AE, including lymphocytic perivascular infiltration and neuronal central chromatolysis in the brain and spinal cord, as well as gliosis in the cerebellar molecular layer. The AE virus was detected by reverse-transcriptase PCR in the brain homogenate from three cases and peripheral nerves in one case. Additionally, the AE virus was isolated in specific-pathogen-free (SPF) embryonated eggs from brain tissue pool samples. Other avian viral infections capable of causing encephalitis, including avian paramyxoviruses, avian influenza virus (AIV), West Nile virus (WNV), eastern equine encephalitis virus (EEEV), and western equine encephalitis virus (WEEV), were ruled out by attempting virus isolation and molecular procedures.

Clinical observations and diagnostic procedures carried out to elucidate the cause of high mortality in 2–8-wk-old ornamental ducks (mandarin, wood, falcated, and silver teal ducks) are described. At necropsy, ducklings showed general pallor of skeletal and heart muscles, subcutaneous gelatinous transudates, pericarditis, ascites, and severe edema and hyperemia of lungs. Histopathologic examination revealed that the most important changes were located in the crop, bursa of Fabricius, and lungs with presence of amorphic basic intracytoplasmic inclusions. No bacteria or fungi could be detected from affected organs and ascitic fluid. Viral diagnosis included molecular detection for the presence of goose parvovirus (GPV), circovirus, avian influenza, herpesviruses, paramyxovirus, reovirus, and polyomavirus. Both GPV and circovirus could be detected by real-time PCR and nested broad-spectrum PCR, respectively. Phylogenetically, full-length nucleotide sequence of GPV showed a close similarity ranging from 95.6% to 97.9% with European and Asian pathogenic GPV. On the other hand, the detected circovirus showed nucleotide identity of 90% to 98% with goose circoviruses (GoCVs). This is the first report of GoCVs and GPV in ornamental ducks. The concurrence of GPV and GoCV infections is thought to contribute to the high mortality.

Over 4 years, only two known cases of fluke invasions were diagnosed in racing pigeons (Columba livia) originating from different regions of Poland. In both cases, the invasion was characterized by a very high mortality (approximately 70%), and the source of the infestation was snails of the Lymnaeidae family eaten by pigeons. Fluke invasions in pigeons are extremely rare and to date have not been described in Poland. Therefore, the occurrence of the symptoms of hemorrhagic diarrhea and sudden deaths of either adult pigeons or nestlings were suspected to be associated with poisoning. Autopsy revealed an invasion of flukes causing hemorrhagic enteritis. Renal failure and spleen atrophy were also found in the birds. Using molecular biology techniques, infestation with the fluke Echinostoma revolutum was determined in the second case.

Runting stunting syndrome (RSS) is a disease condition that affects broilers and causes impaired growth and poor feed conversion because of enteritis characterized by pale and distended small intestines with watery contents. The etiology of the disease is multifactorial, and a large variety of viral agents have been implicated. Here we describe the detection and isolation of an infectious bronchitis virus (IBV) -like coronavirus from the intestines of a flock of 60,000 14-day-old brown/red broiler chicks. The birds showed typical clinical signs of RSS including stunting and uneven growth. At necropsy, the small intestines were pale and distended with watery contents. Histopathology of the intestines revealed increased cellularity of the lamina propria, blunting of villi, and cystic changes in the crypts. Negative stain electron microscopy of the intestinal contents revealed coronavirus particles. Transmission electron microscopy of the intestine confirmed coronavirus in the cytoplasm of enterocytes. Using immunohistochemistry (IHC), IBV antigen was detected in the intestinal epithelial cells as well as in the proventriculus and pancreas. There were no lesions in the respiratory system, and no IBV antigen was detected in trachea, lung, air sac, conjunctiva, and cecal tonsils. A coronavirus was isolated from the intestine of chicken embryos but not from the allantoic sac inoculated with the intestinal contents of the broiler chicks. Sequencing of the S1 gene showed nucleic acid sequence identities of 93.8% to the corresponding region of IBV California 99 and of 85.7% to IBV Arkansas. Nucleic acid sequence identities to other IBV genotypes were lower. The histopathologic lesions in the intestines were reproduced after experimental infection of specific-pathogen-free chickens inoculated in the conjunctiva and nares. Five days after infection, six of nine investigated birds showed enteritis associated with IBV antigen as detected by IHC. In contrast to the field infection, birds in the experimental group showed clear respiratory signs and lesions in the upper respiratory tract. The results suggest a broader tissue tropism of this isolate, which might be related to the mutations in the S1 gene.

Highly pathogenic avian influenza (HPAI) virus of the H5N8 subtype was isolated from a young ostrich in South Korea in March 2014. Clinical signs characterized by anorexia, depression, and signs of nervousness were observed. The isolated A/ostrich/Korea/H829/2014 (H5N8) virus had a cleavage site motif containing multiple basic amino acids, typical of HPAI virus. The phylogenetic tree of the hemagglutinin gene of the H5 HPAI virus showed that this ostrich H5N8 virus belongs to clade 2.3.4.4 viruses together with H5N8 strains isolated from ducks and wild birds in South Korea in 2014. Pathologically, redness of pancreas, enlargement and hemorrhage of spleen, friability of brain, and hydropericardium were prominently found. Histologic legions were observed in pancreas, spleen, liver, lung, heart, and brain, and influenza A nucleoproteins were detected in the same organs by immunohistochemistry. Other ostriches farmed together in open camps were not infected with HPAI virus based on the serologic and virologic tests. The findings indicate that ostriches are susceptible to H5N8 HPAI virus, but this virus does not spread efficiently among ratites.

A mortality episode of endemic and endangered psittacine birds from the genera Ara and Amazona occurred during January 2015. The birds were housed in a management unit for wildlife conservation that receives wild-caught birds from illegal trade. In total, 11 (57%) adult birds of different origins that shared these accommodations died. Only four of them were sent for diagnosis. The main lesions found at necropsy were consistent with those described previously for avian chlamydiosis; the presence of Chlamydiaceae was confirmed through immunofluorescence and amplification with further sequencing of the 16S ribosomal RNA gene by using hepatic tissue. Due to the lack of specific diagnostic tools on primary psittacine diseases, the pathogenic effects of systemic, respiratory, or enteric infections with high mortality rates remain unknown in Mexico. In this study, specific molecular identification of avian chlamydiosis was performed using a nested PCR on liver tissues, as well as choanal and cloacal swab samples, confirming the presence of Chlamydia psittaci in all of them. In addition, it was possible to obtain the ompA gene sequence from processed clinical samples, thereby allowing us to determine that the A genotype was affecting these birds. Although this genotype is the most commonly found worldwide in psittacine birds, this case report describes the first avian chlamydiosis outbreak affecting critically endangered and endemic psittacines subjected to reintegration programs in Mexico. Consequently, this study demonstrates the necessity of more exhaustive biosecurity strategies because other pathogens may be present and should be assessed, especially in highly threatened birds, before releasing them into their habitats.