Avian Encephalomyelitis Virus Research Articles

Module-combinatorial design and screening of multifunctional polymers based on polyaspartic acid for DNA delivery

It is crucial to develop non-viral gene vectors that can efficiently and safely transfect plasmid DNA into cells. Low transfection efficiency and high cytotoxicity of cationic polymers hinder their application as gene carriers. Modification of cationic polymers has emerged as an attractive strategy for efficient and safe nucleic acids delivery. In this study, a simple and rapid method is developed to synthesize a series of multifunctional polymers by utilizing biodegradable polyaspartic acid as the backbone and modifying it with three modules. This one-component polymer possesses capabilities for nucleic acid condensation, cellular uptake, and endosomal escape. Polymers containing imidazole, triazole, or pyridine group exhibited promising transfection activity. Substituted with dodecylamine or 2-hexyldecan-1-amine enhance cellular uptake and subsequent transfection. Furthermore, the influence of ionizable amine side chains on gene delivery is investigated. Two optimal polymers, combined with the avian encephalomyelitis virus (AEV) plasmid vaccine, induced robust specific antibody responses and cellular immune responses in mice and chickens. Through module-combination design and screening of polyaspartamide polymers, this study presents a paradigm for the development of gene delivery vectors.

The full genome characterization of avian encephalomyelitis virus, Iran: a vertical transmission case.

Avian encephalomyelitis (AE) is an important infectious poultry disease worldwide that is caused by avian encephalomyelitis virus (AEV). The causative virus can be transmitted both horizontally and vertically. In the present study, an AEV suspected outbreak with typical neurological signs occurred in broilers. Histopathological examination, RT-PCR assay and full genome sequencing were applied to confirm the presence of AEV. Phylogenetic analysis of the full genome sequence showed that the detected AEV strain at 7055 nucleotide length is classified in cluster I and is closely related to vaccinal USA and China originated isolates. Although, the outbreaks of AE in progeny of vaccinated breeders have been reported previously, the source of infection was unknown. Based on the results obtained in this study, the outbreaks are vaccine-originated. This study provides the first whole genome analysis of AEV from Iran and reveals that the AEV possesses a hepatitis C virus-like internal ribosome entry site.

Occurrences of avian encephalomyelitis virus in naturally infected chicks in Saudi Arabia's Eastern Province.

A neurological infectious viral disease, avian encephalomyelitis was initially discovered in 2-week-old commercial chicks in 1930 and classified as a neurotropic viral disease. A neurological outbreak caused by avian encephalomyelitis virus (AEV) in young chicks was first reported in Al-Ahsa in the Kingdom of Saudi Arabia (KSA) in 2010. The aim of this article is to examine the AEV in KSA, Al-Ahsa Province. Gizzard, proventriculus, cerebrum, cerebellum, and medulla oblongata tissue samples were collected from infected chicks for histopathology test and molecular identification. Infected chicks showed neurological signs particularly incoordination, mild head and neck tremors, stretching of legs, and lameness. The average morbidity and mortality rates were 35% and 10%, respectively. At necropsy, no obvious identifiable macroscopic lesions were found in the infected chicks. Nonsuppurative encephalomyelitis was found histopathologically in the central nervous system, mainly in the cerebral molecular layer. Microscopic lesions in the proventriculus showed masses of heavy numbers of small lymphocytes within the muscular layer. RT-PCR followed by sequence analysis revealed that The KSA strain (KJ939252) is intimately related to chicken European strains from Poland (KC912695) and the United Kingdom (AJ225173) with identity 99.6% than Chinese strains (AY225319, AY517471, and AY275539) with identity ranged between 94.6% and 95%. The phylogenetic tree analysis showed that the KSA strain is grouped in a similar clade with chicken European strains. The pattern of disease findings was typical of vertically transmitted AEV. The spread of AEV in Saudi Arabia is most likely due to the trade of birds and bird products with European countries.

Research Note: Pathogenetic characteristics of avian encephalomyelitis virus in Guangdong and Jiangxi Provinces, China

In recent years, the infection rate of avian encephalomyelitis virus (AEV) infection in chickens has risen significantly, seriously endangering the development of the chicken industry. In order to study the current epidemiological status of AEV in China as well as the genetic and evolutionary patterns of the virus, we conducted a survey and genomic analysis of chicken AEV. The results showed that 46.26% (136/294) of the tissue samples tested (n = 294) were positive for AEV, with the highest positivity rate of 62.24% (61/98) among tissue samples from chickens aged 13 to 18 wk. The complete genomes of 2 representative AEV strains were determined, and the VP1 evolutionary tree results revealed that the 2 representative strains belonged to a novel AEV strain. Multiple alignment analysis showed that the ORF1 genes of the 2 representative strains differed by 82.3 to 99.9% at the amino acid level compared with the reference AEV strain, and the mutations at the key amino acid loci of VP2 and VP3 were the same as those in the chick embryo-adapted strain. The analysis makes up for the molecular epidemiological data and genetic variation of the 2 representative strains. The analysis makes up for the molecular epidemiological data and genetic variation of AEV and provides a basis for further understanding the spread of AEV in China.

Application of metagenomics for diagnosis of broilers displaying neurological symptoms

BackgroundThirty-two-day-old broiler chickens at a farm located in northwestern South Korea displayed adverse neurological symptoms including limping, lying down, and head shaking. Approximately 2.1% of chickens died or were culled due to severe symptoms. Five carcasses were submitted to the Avian Disease Division of the Animal and Plant Quarantine Agency (APQA) for disease diagnosis.ResultsBroilers displayed severe pericarditis and perihepatitis associated with gross lesions. Broilers also displayed microscopic lesions in the cerebrum and in the granular layer of the cerebellum, which were associated with multifocal perivascular cuffing and purulent necrosis in the cerebrum, and severe meningitis with heterophil and lymphocyte infiltration. Staphylococcus spp. were identified in the liver and heart using bacteriological culture. PCR/RT-PCR assays revealed that broilers were negative for avian Clostridium botulinum, Newcastle disease virus, and avian encephalomyelitis virus. Bacterial and viral metagenomic analysis of brain sample further revealed the presence of Pseudomonas spp. and Marek’s disease virus, which are known etiological agents of chicken meningoencephalitis.ConclusionsThis study reports a diagnostic analysis of gross and histopathological lesions from 32-day-old broilers displaying unique neurological symptoms that revealed the presence of the several neurological diseases including meningoencephalitis. The causative agents associated with meningoencephalitis of broilers that had not been identified by routine diagnostic methods could be diagnosed by metagenomics, which proves the usefulness of metagenomics as a diagnostic tool for unknown neurological diseases in broilers.

Avian encephalomyelitis virus in backyard chickens.

Avian viral diseases usually cause high economic losses because of high morbidity and mortality and poor growth. The rearing of chickens in backyards could have an important role in the spread of certain diseases, particularly those of viral origin. Infected birds might be prone to many viral infections for several reasons, including a lack of vaccination programs, the mixing of different bird species in the same location, and the close interactions of these birds with wild and migratory birds carrying various pathogens. This study aimed to conduct serological surveillance of avian encephalomyelitis virus (AEV) in some backyard chickens in the eastern region of Saudi Arabia. Serum samples (n = 368) were collected from domestic chickens reared in 10 backyards in the Eastern Province of Saudi Arabia. None of the domestic birds in these 10 backyards were vaccinated against the virus. In addition, 78 serum samples were collected from free-ranging birds belonging to Columbidae, such as pigeons and doves, in common areas near the domestic backyards. We tested these sera for specific antibodies against AEV. Our results revealed seroconversion to AEV among the examined chickens (14.6%). None of the tested pigeons and doves displayed seroconversion to AEV. Seroconversion of these non-vaccinated birds against AEV was suggestive of a recent natural infection by this virus. Further studies with a large number of birds are required to molecularly characterize the circulating strains of this virus in this area.

DIVA MOLECULAR DETECTION OF SUSPECTED CASE OF NEWCASTLE AND ENCEPHALOMYLITIS DISEASE IN LAYERS

The aim of the study investigates the molecular diagnosis between (the Avian encephalomyelitis virus and Newcastle disease virus and differentiating the infected from vaccinated animals (DIVA) strategies. The 1st detection by RT-PCR for(PMV-1) as primary detection then, the samples were prepared as in FTA card Whatman® and sent to (AniCon Labs) Germany to detect by one step RT-PCR (DIVA technique) between NDV and AEV genotype. The result of the molecular investigation as primary detection revealed three samples were positive with Ct values (34.0, 26.6, and 35.8) respectively. Then, in the detection of AEV the result showed all samples were negative. finally, by using (DIVA) with two primers to detect general (PMV1-9) by (M-gene) all samples were positive at Ct values (20.8, 20.1, and 25.2) respectively, and then path type by (F gene) to differentiate infected from vaccinated samples also all samples were positive with Ct values (23.8, 23.3 and 23.1) respectively. the study concluded that the samples can differentiate between AEV and NDV by using DIVA strategies to find pathotypes and differentiating between infected and vaccinal isolate based on certain amino acids in the primers.

Virome Profiling of Chickens with Hepatomegaly Rupture Syndrome Reveals Coinfection of Multiple Viruses.

Liver diseases seriously challenge the health of chickens raised on scaled farms and cause tremendous economic losses to farm owners. The causative agents for liver diseases are still elusive, even though various pathogens, such as the hepatitis E virus, have been reported. In the winter of 2021, a liver disease was observed on a chicken farm in Dalian, China, which increased chicken mortality by up to 18%. We conducted panvirome profiling of the livers, spleens, kidneys, and recta of 20 diseased chickens. The viromic results revealed coinfection of multiple viruses, including pathogenic ones, in these organs. The viruses were highly identical to those detected in other provinces, and the vaccine and field strains of avian encephalomyelitis virus (AEV) and chicken infectious anemia virus (CIAV) cocirculated on the farm. In particular, the liver showed higher abundance of AEV and multiple fowl adenoviruses than other organs. Furthermore, the liver also contracted avian leukemia virus and CIAV. Experimental animals with infected liver samples developed minor to medium lesions of the liver and showed a virus abundance profile for AEV across internal organs similar to that in the original samples. These results suggest that coinfection with multiple pathogenic viruses influences the occurrence and development of infectious liver disease. The results also highlight that strong farm management standards with strict biosafety measures are needed to minimize the risk of pathogenic virus introduction to the farm.

The brain-specific upregulation of CARD11 in response to avian brain-neurotropic virus infection serves as a potential biomarker

Avian neurotropic viruses are critical problems in poultry industry causing severe central nervous system (CNS) damage with neuroinvasive and neurovirulence properties. Biomarker of neurotropic viral intracranial invasion is of great application value for the diagnosis, but that of avian neurotropic viruses remains elusive. Previously, we found that chicken caspase recruitment domain family, member 11 (CARD11) was only upregulated in virulent Newcastle disease virus-infected chickens and in chicken primary neuronal cells. In this study, CARD11 was systemically expressed in chickens and pigeons detected by absolute qPCR and immunohistochemical (IHC) assay. After virus challenging, only avian neurotropic viruses (avian encephalomyelitis virus [AEV] and pigeon paramyxovirus type 1 [PPMV-1]) except Marek's disease virus (MDV) can invade brain and cause pathological changes. The relative mRNA expression of CARD11 was brain-upregulated in AEV- or PPMV-1-infected animals, rather than MDV and non-neurotropic viruses (fowl adenovirus serotype 4 [FAdV-4] and infectious bronchitis virus [IBV]). Similarly, the protein expression of CARD11 was only upregulated in the cerebra and cerebella infected by avian brain-neurotropic virus using IHC assay. And there were no correlations between the change level of CARD11 and viral load. Our preliminary data suggested that avian CARD11 may be a potential brain biomarker for avian brain-neurotropic virus invasion.

Molecular-based monitoring of live vaccines in dust samples from experimental and commercial chicken flocks and its potential use as a screening test

Despite the high cost of vaccination programmes, conventional methods to evaluate vaccine uptake are often impractical and costly. More recently, molecular-based testing of poultry dust has been used to monitor the “take” of Marek's disease virus and infectious laryngotracheitis virus (ILTV) live vaccines. This study aimed to provide proof-of-concept for detecting other poultry pathogens by using molecular detection of vaccine microorganisms in poultry dust of vaccinated flocks. Dust and choanal cleft and cloacal swabs were collected from chickens vaccinated against avian encephalomyelitis virus (AEV), fowlpox virus (FPV), Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) using live vaccines in an experimental flock. Dust samples were collected weekly from 5 commercial breeder or layer flocks from day-old up to 25 weeks of age. These flocks were vaccinated against Newcastle disease virus (NDV), infectious bronchitis virus (IBV), infectious bursal disease virus (IBDV), ILTV, fowl adenovirus (FAdV), MG and MS. Samples were tested for nucleic acids of these microorganisms by PCR or reverse transcriptase PCR. Genomes of all targeted vaccines were detected in dust samples from the experimental and commercial flocks except for FPV, which was detected only in the experimental flock. FAdV was detected in unvaccinated commercial flocks. These findings suggest that PCR detection of target organisms in dust samples has potential as a relatively simple and inexpensive population-level test to monitor vaccine take and/or pathogen status in chicken flocks. Further studies comparing the detection of each of these microorganisms in poultry dust with individual birds samples are required to validate this approach.

Seroprevalences of specific antibodies against avian pathogens in free-ranging ring-necked pheasants (Phasianus colchicus) in Northwestern Germany

Infectious diseases in captive pheasants (Phasianus colchicus) are well known, but there is a lack of knowledge about occurrence and distribution of pathogens in free-ranging pheasants in Germany. We investigated 604 sera from hunted pheasants and 152 sera from wild caught pheasants between 2011 to 2015, with the aim to determine the prevalence of specific antibodies against different viruses: Avian influenza virus (AIV) of subtypes H5, H7, H9, paramyxovirus type 1 (PMV-1), avian encephalomyelitis virus (AEV), infectious bursitis disease virus (IBDV), infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV), avian metapneumovirus (aMPV) and Salmonella sp., Mycoplasma synoviae (MS) and Mycoplasma gallisepticum (MG). In addition, 178 caeca were investigated for Histomonas meleagridis. The study reveals an ongoing circulation of IBV in the wild pheasant population during the study. Also high seroprevalences of specific antibodies against aMPV depending on the area and a strong increase in prevalence of IBDV antibodies in sera of pheasants in Lower Saxony were detected. ILTV antibody prevalences differed between areas and AEV antibody detection differed between years significantly, whereas specific antibodies against PMV-1 could not be detected and antibodies against AIV-H5, -H7 and -H9 and Mycoplasma spp. were detected in very few cases.

Characterization of a fowl adenovirus 9 (FAdV-9) early promoter and its application in generating dual expression FAdV-9s

The CMV immediate early promoter from the EGFP expression plasmid pEGFP-N1 was replaced with the very left end of the fowl adenovirus 9 (FAdV-9) genome (ntds 73−574) to demonstrate and delineate the promoter function of this sequence. Expression of an EGFP ORF which replaced ORF1 and ORF2 demonstrated that the native promoter can drive down stream foreign gene expression. Replacement of ORF1 and ORF2 with a bicistronic cassette, incorporating a 493 bp IRES from an Ontario strain of avian encephalomyelitis virus (AEV) separating an EGFP ORF and mCherry ORF allowed for expression of both ORFs from a recombinant FAdV. These results provide an additional platform for multivalent vaccines development based on a native FAdV-9 promoter and an avian virus IRES.

Structural Analysis of Avian Encephalomyelitis Virus Polyprotein for Development of Multi Epitopes Vaccine Using Immunoinformatics Approach

Avian Encephalomyelitis (AE) is the disease caused by avian encephalomyelitis virus (AEV). The disease mainly affects young birds nervous system worldwide causing high morbidity and variable mortality rate in chicks and noticed egg dropping and hatchability in mature hens. Vaccination is the only way to control AEV infection since there is no treatment yet to the avian encephalomyelitis. This study aimed to use immunoinformatics approaches to predict multi epitopes vaccine from the AEV polyprotein that could elicit both B and T cells. The vaccine construct comprises 482 amino acids obtained from epitopes predicted against B and T cells by IEDB server, adjuvant, linkers and 6-His-tag. The chimeric vaccine was potentially antigenic and nonallergic and demonstrated thermostability and hydrophilicity in protparam server. The solubility of the vaccine was measured in comparison to E. coli proteins. The stability was also assessed by disulfide bonds engineering to reduce the high mobility regions in the designed vaccine. Furthermore molecular dynamics simulation further strengthen stability of the predicted vaccine. Tertiary structure of the vaccine construct after prediction, refinement was used for molecular docking with chicken alleles BF2*2101 and BF2*0401 and the docking process demonstrated favourable binding energy score of -337.47 kcal/mol and -326.87 kcal/mol, respectively. Molecular cloning demonstrated the potential clonability of the chimeric vaccine in pET28a(+) vector. This could guarantee the efficient translation and expression of the vaccine within suitable expression vector.

First report on the seroprevalence of avian encephalomyelitis virus antibody in Sonali (cross-bred) chickens in Bogura, Bangladesh.

Objectives:The study intended to detect the presence and distribution of avian encephalomyelitis virus (AEV)-specific antibodies in Sonali (cross-bred) parent chickens regarding farm location, flock size, and age in Bogura district of Bangladesh, a Sonali chicken belt.Materials and methods:A total of 275 Sonali parent chickens’ blood samples were collected randomly from 39 flocks during laying age with a healthy and non-vaccination history against AEV. Blood samples were collected aseptically from the wing veins of chickens using 3-ml syringes and sera were separated. Then, the sera were transferred to the laboratory by maintaining a cool chain. Indirect enzyme-linked immunosorbent assay was used to detect the specific antibodies against AEV present in the sera samples.Results:Overall, 70.18% of the chickens were found seropositive for AEV antibodies. Based on the location, the highest seropositivity was recorded in Bogura Sadar [91.30%, confidence intervals (CI) 79.21%–97.58%], and the lowest was in the Adomdighi sub-district (45.45%, CI 29.49%–63.08%). For flock size, AEV seropositivity was significantly (p < 0.05) higher in the large flock (82.22%, CI 72.74%–89.48%). Regarding age groups, the seropositivity of AEV was significantly (p < 0.05) increased with chickens’ age. Higher seropositivity was noted in chickens aged >51 weeks (89.32%, CI 81.69%–94.55%).Conclusion:The results indicate that AEV is circulating in the environment, and chickens were exposed to the field strain of AEV. To the best of our knowledge, this is the first report on AEV in chickens in Bangladesh. Proper vaccination and standard farm biosecurity practice could minimize AEV infection in chickens. A detailed epidemiology study, detection, and characterization of the AEV would be essential for effective AEV infection control.

Health status of free-ranging ring-necked pheasant chicks (Phasianus colchicus) in North-Western Germany.

Being a typical ground-breeding bird of the agricultural landscape in Germany, the pheasant has experienced a strong and persistent population decline with a hitherto unexplained cause. Contributing factors to the ongoing negative trend, such as the effects of pesticides, diseases, predation, increase in traffic and reduced fallow periods, are currently being controversially discussed. In the present study, 62 free-ranging pheasant chicks were caught within a two-year period in three federal states of Germany; Lower Saxony, North Rhine-Westphalia and Schleswig-Holstein. The pheasant chicks were divided into three age groups to detect differences in their development and physical constitution. In addition, pathomorphological, parasitological, virological, bacteriological and toxicological investigations were performed. The younger chicks were emaciated, while the older chicks were of moderate to good nutritional status. However, the latter age group was limited to a maximum of three chicks per hen, while the youngest age class comprised up to ten chicks. The majority of chicks suffered from dermatitis of the periocular and caudal region of the head (57-94%) of unknown origin. In addition, intestinal enteritis (100%), pneumonia (26%), hepatitis (24%), perineuritis (6%), tracheitis (24%), muscle degeneration (1%) and myositis (1%) were found. In 78% of the cases, various Mycoplasma spp. were isolated. Mycoplasma gallisepticum (MG) was not detected using an MG-specific PCR. Parasitic infections included Philopteridae (55%), Coccidia (48%), Heterakis/Ascaridia spp. (8%) and Syngamus trachea (13%). A total of 8% of the chicks were Avian metapneumovirus (AMPV) positive using RT-PCR, 16% positive for infectious bronchitis virus (IBV) using RT-PCR, and 2% positive for haemorrhagic enteritis virus (HEV) using PCR. All samples tested for avian encephalomyelitis virus (AEV), infectious bursal disease virus (IBDV) or infectious laryngotracheitis virus (ILTV) were negative. The pool samples of the ten chicks were negative for all acid, alkaline-free and derivative substances, while two out of three samples tested were positive for the herbicide glyphosate. Pheasant chick deaths may often have been triggered by poor nutritional status, probably in association with inflammatory changes in various tissues and organs as well as bacterial and parasitic pathogens. Theses impacts may have played a major role in the decline in pheasant populations.

Circulation of Immunosuppressive Viruses and Avian Encephalomyelitis Virus in Backyard Chicken Flocks

The objective of this study was to evaluate the circulation of Chicken Anemia Virus (CAV), Infectious Bursal Disease Virus (IBDV), Avian Reovirus (ARV) and Avian Encephalomyelitis virus (AEV) in properties of backyard chickens and carry out an epidemiological analysis. We evaluated 200 samples of chickens from 19 backyard chicken property. Only one property (P10) did not present serological titers for the diseases evaluated. This property is close to industrial farms as well as the other properties, however, P10 remained a few years without the breeding of chicks and these were the first poultry to be housed on site. This reinforces the importance of the fallow period for poultry production. The prevalence of virus-seroreactive birds was 78% (156/200), 64.5% (129/200), 78% (156/200), 78% (156/200) for CAV, IBDV, ARV and, EA, respectively. All the free-range farms studied are within a radius of 500 meters to 6 Km away from some establishments of industrial poultry. There was a correlation between serological titers for CAV and the frequency of disease in poultry (r = 0.6178). In places where birds are frequently sick, 30.76% reported that the disease occurs in chicks, 30.76% in broilers, 23.07% in broiler chickens and 7.69% in birds of all ages. Birds get sick more often in the summer period. The owners reported that the most common signs of disease were respiratory signs (snoring and nasal discharge) (46.15%), diarrhea (30.76%), and paralysis of wings and/or paws (38.46%). There was a correlation between the presence of untreated water in the property and serological titers for ARV (r = 0.5576). This report draws attention not only to high serological prevalence for the viruses studied but also important epidemiological aspects of backyard chicken diseases that may indirectly influence the industrial production.

Field safety and efficacy of a unique live virus vaccine for controlling avian encephalomyelitis and fowlpox in poultry.

Background and Aim:Infection of commercial poultry with avian encephalomyelitis (AE) and fowlpox (FP) virus causes heavy economic loss in endemic areas. Although vaccines are routinely used to control these two diseases, the problem still persists almost all over the world. This study aimed to evaluate safety and efficacy of a unique AE + FP + pigeon pox (PP) live virus vaccine in layer-type chickens under both laboratory and field conditions.Materials and Methods:The study was conducted using 289 specific-pathogen-free (SPF) chickens under the laboratory conditions and 185,648 commercial layer-type chickens under field conditions. In two consecutive laboratory trials, 8-week-old SPF chickens were vaccinated with the AE + FP + PP live virus vaccine through wing web route and challenged against virulent strains of FP and AE viruses at 3-week post-vaccination (WPV). Challenged chickens were observed for disease protection for 10-21 days. For field safety trials, commercial layer-type chickens in three different geographical areas in the USA were vaccinated with the AE + FP + PP vaccine and observed daily up to 21 days for vaccine “take,” adverse reactions, and mortality.Results:The vaccine was found safe and efficacious under both laboratory and field conditions. Vaccine “take” and protection against FP challenge were 100%. Average protection against AE challenge was 97%. Mean AE enzyme-linked immunosorbent assay (ELISA) antibody titer in the field vaccinated chickens was >1200 at 10 WPV. Average daily post-vaccination mortality in the field vaccinated chickens was 0.04%. So far, more than 400 million chickens in the USA have been vaccinated with this vaccine. No vaccine-associated adverse reactions, other safety issues, or immunity breakdown cases in the vaccinated flocks due to field virus infection have been reported.Conclusion:This unique vaccine containing AE, FP, and PP viruses in a single preparation was found to be safe and efficacious in controlling the diseases caused by the virulent field strains of AE and FP. Besides being safe and efficacious, this vaccine also offered distinct advantages over the traditional vaccination practices in controlling these two diseases in poultry.

Detection of avian encephalomyelitis virus in chickens in Japan using RT-PCR.

A reverse transcription-polymerase chain reaction (RT-PCR) method was developed for broadly detecting the avian encephalomyelitis virus (AEV). The new primers were based on conserved sequences of the 5’-untranslated region of AEV, because the virus was not detected using previous reported RT-PCR. By applying this method to the chicken samples with suspected AEV infection in Japan, we successfully obtained PCR products of the predicted size from all samples, and we confirmed the presence of AEV via sequence analysis.

Assessing the efficacy of a live vaccine against avian encephalomyelitis virus.

Avian encephalomyelitis virus (AEV) causes typical neurological symptoms in young chicks and a transient drop in egg production and hatchability in adult laying birds, resulting in huge economic losses in the poultry industry. An effectiveway to control and prevent this disease is vaccination of the flocks. Here, we assessed the efficacy of the live vaccine candidate strain GDt29 against avian encephalomyelitis virus. The GDt29 strain has low virulence, was confirmed safe, and showed no signs of pathogenicity. High titers of AEV-specific antibodies were detected in GDt29-vaccinated hens (S/P > 3.0) and their progeny (S/P > 2.0). Moreover, the eggs of GDt29-vaccinated hens with high levels of maternal antibodies were hatched successfully regardless of challenge with a heterologous AEV strain, and the GDt29 attenuated vaccine showed higher protective efficacy against AEV than the commercial vaccine. Furthermore, contact-exposed chicks bred with GDt29-vaccinated birds generated high titers against AE virus (S/P > 2.8). Collectively, our studies are proof of the principle that GDt29 might be an ideal vaccine candidate to prevent AEV infection, and they highlight the utility of using a live vaccine against AEV.

Investigation into diseases in free-ranging ring-necked pheasants (Phasianus colchicus) in northwestern Germany during population decline with special reference to infectious pathogens.

The population of ring-necked pheasants (Phasianus colchicus) is decreasing all over Germany since the years 2008/2009. Besides impacts of habitat changes caused by current rates of land conversion, climatic influences or predators, a contribution of infectious pathogens needs also to be considered. Infectious and non-infectious diseases in free-living populations of ring-necked pheasants have been scarcely investigated so far. In the present study, carcasses of 258 deceased free-ranging pheasants of different age groups, predominantly adult pheasants, collected over a period of 4 years in the states of Lower Saxony, North Rhine–Westphalia and Schleswig-Holstein, were examined pathomorphologically, parasitologically, virologically and bacteriologically, with a focus set on infectious pathogens. A periocular and perinasal dermatitis of unknown origin was present in 62.3% of the pheasants. Additional alterations included protozoal cysts in the skeletal musculature (19.0%), hepatitis (21.7%), enteritis (18.7%), gastritis (12.6%), and pneumonia (11.7%). In single cases, neoplasms (2.6%) and mycobacteriosis (1.7%) occurred. Further findings included identification of coronaviral DNA from trachea or caecal tonsils (16.8%), siadenoviral DNA (7.6%), avian metapneumoviral RNA (6.6%), and infectious bursal disease viral RNA (3.7%). Polymerase chain reaction (PCR) on herpesvirus, avian influenza virus (AIV), paramyxovirus type 1 (PMV-1), avian encephalomyelitis virus (AEV), and chlamydia were negative. Based on the present results, there is no indication of a specific pathogen as a sole cause for population decline in adult pheasants. However, an infectious disease can still not be completely excluded as it may only affect reproduction effectivity or a certain age group of pheasants (e.g., chicks) which were not presented in the study.