Prevalence of equine parvovirus-hepatitis in healthy broodmares in Ontario, Canada.

Equine parvovirus-hepatitis (EqPV-H) was first reported from the serum and liver tissue of a horse diagnosed with Theiler's disease in the United States in 2018. Theiler's disease, also known as equine serum hepatitis, is a severe hepatitis with fulminant hepatic necrosis. The disease has most frequently been reported following the administration of equine-origin biological products; however, it has also been reported in in-contact horses with no prior biologic administration. EqPV-H has been detected in clinically healthy horses in North America (USA, Canada), Europe (Germany, Austria, Slovenia), Asia (China, South Korea), and South America (Brazil). Previous prevalence studies conducted worldwide have shown the presence of EqPV-H DNA in serum or plasma ranging from 3.2 to 19.8%. This study investigated the prevalence of EqPV-H DNA in 170 healthy broodmares of various breeds located on 37 farms in southern Ontario, Canada. The occurrence of EqPV-H infection was determined by quantitative PCR for EqPV-H DNA in serum samples. The effects of age, breed, season, pregnancy status, and equine herpesvirus-1 (EHV-1) vaccination history on EqPV-H status were also investigated. There was a prevalence of 15.9% (27/170) with viral loads of EqPV-H ranging from detectable to 2900 copies/mL. Statistical analysis showed that increasing age was a significant factor in the detection of EqPV-H DNA. Neither breed, season, pregnancy status, nor EHV-1 vaccination history was significant in predicting EqPV-H infection status.

L'hépatite à parvovirus équin (EqPV-H) a été signalée pour la première fois à partir du sérum et du tissu hépatique d'un cheval diagnostiqué avec la maladie de Theiler aux États-Unis en 2018. La maladie de Theiler, également connue sous le nom d'hépatite sérique équine, est une hépatite sévère avec nécrose hépatique fulminante. La maladie a été le plus souvent rapportée à la suite de l'administration de produits biologiques d'origine équine; cependant, il a également été signalé chez des chevaux en contact sans administration préalable de produit biologique. EqPV-H a été détecté chez des chevaux cliniquement sains en Amérique du Nord (États-Unis, Canada), en Europe (Allemagne, Autriche, Slovénie), en Asie (Chine, Corée du Sud) et en Amérique du Sud (Brésil). Des études de prévalence antérieures menées dans le monde entier ont montré la présence d'ADN EqPV-H dans le sérum ou le plasma allant de 3,2 à 19,8 %. Cette étude a examiné la prévalence de l'ADN EqPV-H chez 170 poulinières en bonne santé de différentes races situées dans 37 fermes du sud de l'Ontario, au Canada. La survenue d'une infection par EqPV-H a été déterminée par PCR quantitative pour l'ADN d'EqPV-H dans des échantillons de sérum. Les effets de l'âge, de la race, de la saison, de l'état de grossesse et des antécédents de vaccination contre l'herpèsvirus équin-1 (EHV-1) sur le statut EqPV-H ont également été étudiés. Il y avait une prévalence de 15,9 % (27/170) avec des charges virales d'EqPV-H allant de détectable à 2900 copies/mL. L'analyse statistique a montré que l'augmentation de l'âge était un facteur significatif dans la détection de l'ADN EqPV-H. Ni la race, ni la saison, ni l'état de grossesse, ni les antécédents de vaccination contre l'EHV-1 n'étaient significatifs pour prédire l'état de l'infection par l'EqPV-H.(Traduit par Docteur Serge Messier).

Molecular surveillance of equine parvovirus-hepatitis from oral, nasal, vaginal, and semen specimens collected from horses living in Korea.

Equine parvovirus-hepatitis (EqPV-H) is one of the etiological agents of Theiler's disease, causing fulminant hepatitis; however, its transmission route and pathogenesis remain unclear. In the present study, we aimed to determine EqPV-H shedding in oral/nasal/vaginal swabs or semen samples from horses living in Korea using nested polymerase chain reaction. We then used the data obtained to investigate various risk factors associated with EqPV-H including viral shedding, hepatopathological changes, and genetic diversity. Our data revealed occurrence of EqPV-H shedding in these animals (oral: 3/102 [2.9%]; nasal: 3/102 [2.9%]; semen: 1/9 [11.1%]) and identified that both age and country of foaling were significantly associated with EqPV-H shedding (p < .05). In addition, we noted that one of the newly isolated strains clustered separately from the other strains in the phylogenetic tree, revealing unique nucleotide and amino acid substitutions. This is a field surveillance study providing evidence of natural and venereal shedding of EqPV-H and describing its presence in both oral/nasal fluids and semen. This epidemiological and clinical analysis may help specify the clinicopathological features of EqPV-H and facilitate the development of novel disease prevention strategies.

Duck hepatitis A virus prevalence in mainland China between 2009 and 2021: A systematic review and meta-analysis.

Duck hepatitis A virus (DHAV) is a single-strand positive-sense small RNA virus that causes high mortality in ducklings. In recent years, the incidence of DHAV-3 subtype has been increasing in China, leading to great economic losses to the duck-breeding industry. We investigated the incidence and mortality rates of DHAV in ducks and analysed the seroprevalence of DHAV in mainland China, by meta-analysis. Twenty-six studies published between 2009 and 2021 were retrieved, with a total of 689,549 cases from 14 provinces. Using the DerSimonian-Laird model, DHAV prevalence was estimated with the variance-stabilizing double arcsine transformation. The incidence of DHAV in mainland China was 12 % (95 % confidence interval [CI]: 3-20 %), and the mortality rate was 11 % (95 % CI: 2-19 %), suggesting that the virus was highly virulent and mortality was high. Time analysis showed that DHAV incidence decreased over time. The typing survey showed that strains of DHAV-1 serotype accounted for 38 % (95 % CI: 21-56 %) and strains of DHAV-3 serotype accounted for 49 % (95 % CI: 31-68%) of the tested samples. The decline in the detection rate of DHAV-1 may be due to the widespread use of the DHAV-1 vaccine, which has effectively controlled the DHAV-1 serotype virus. The DHAV-3 vaccine has been on the market for a short time and has no cross protection with DHAV-1, so DHAV-3 accounted for a high proportion of the tested samples. Subgroup analysis of the detection methods showed little difference between PCR and other detection methods.

The history of Theiler's disease and the search for its aetiology.

Theiler's disease (serum hepatitis) may occur in outbreaks or as single cases of acute hepatitis and is often associated with prior administration of equine-origin biologics approximately 4-10 weeks before the onset of clinical signs. Cases have also been described without any prior administration of blood products. The clinical disease has a low morbidity but high mortality and only adult horses are affected. The course of the disease is short, with horses either dying or completely recovering in a few days. Pathology in affected horses is predominantly centrilobular hepatocyte necrosis with mononuclear cell infiltration of the lesser affected periportal regions of the liver. Subclinical cases of the disease also occur. Based on the epidemiology and pathology of the disease, a viral cause, similar to hepatitis B in humans, has long been suspected. This paper reviews both historical and recent findings on Theiler's disease. Reported epidemics of Theiler's disease in the early 1900s are reviewed, along with their similarities to outbreaks of serum hepatitis in humans following yellow fever virus vaccinations in the 1930s and 1940s. Recent metagenomics-based studies to determine the aetiology of Theiler's disease are discussed, along with both clinical and experimental findings supporting equine parvovirus-hepatitis (EqPV-H) as the likely cause of this 100-year-old disease.

Molecular detection of avian hepatitis E virus (Orthohepevirus B) in chickens, ducks, geese, and western capercaillies in Poland.

Orthohepevirus B, commonly known as avian hepatitis E virus (aHEV), causes big liver and spleen disease (BLS) or hepatitis-splenomegaly syndrome (HSS) in chickens. BLS is an emerging disease among chicken flocks in several countries around the world. In our previous studies, serology and molecular biology screening revealed that chicken flocks are widely affected by aHEV in Poland. The present study, which was conducted between 2019 and 2020, aimed to investigate the prevalence of aHEV in chicken flocks and other poultry, including ducks, geese, and turkeys. A total of 307 flocks were examined. In addition, 29 samples from captive wild birds (western capercaillies, Tetrao urogallus) were analyzed. In all the investigated poultry species, except turkeys, the nucleic acid sequence covering part of the ORF1 gene of the aHEV genome was detected (34/336 samples, 10.1%). The infection rate was found to be the highest in broiler breeder chicken flocks (14/40 samples; 35%). Phylogenetic analysis of partial ORF1 gene, which encodes helicase, revealed that the obtained sequences belonged to genotypes 2 and 4, while one belonged to genotype 3. Genotype 2 was detected for the first time in domestic geese and ducks, and genotype 4 was detected for the first time in Poland. The study demonstrated the presence of aHEV among the investigated western capercaillies, suggesting that this species is susceptible to aHEV infections and biosecurity is therefore required in western capercaillie breeding facilities.

Isolation, identification and genome analysis of an avian hepatitis E virus from white-feathered broilers in China.

Avian hepatitis E virus (HEV) is the major causative pathogen of the big liver and spleen disease, hepatitis-splenomegaly syndrome, and hepatic rupture hemorrhage syndrome. Until now, there are 6 different avian HEV genotypes that infect chickens have been reported worldwide. Epidemiologic investigations of the avian HEV demonstrated that avian HEV has been widely spread in China in recent years. In this study, an avian HEV named YT-aHEV was obtained from white-feathered broilers using LMH cells by virus isolation assay in Shandong province, China. The complete genome consists of 6656-nt excluding the poly(A) tail. The isolate was highly similar to the CaHEV strain and segregated into the same branch belonging to avian HEV genotype 3. Indirect immunofluorescence using capsid protein-specific polyclonal antibodies confirmed that YT-aHEV could establish productive infection and replicate stably in LMH cells. Furthermore, an in vivo avian HEV infection model was established successfully in specific pathogen-free chicken embryos by intravenous experiments. In the present study, we demonstrate an avian HEV infection associated with liver lesions of hemorrhage and swelling by LMH cells for the first time in a white-feather broiler flock in China. This research also provides a new diagnosis method for detection of avian HEV, which laid a foundation for the understanding of pathogenicity and molecular biology of this virus for further study.

The diagnosis and molecular epidemiology investigation of avian hepatitis E in Shandong province, China.

BACKGROUND: Avian hepatitis E virus (HEV) is the pathogenic agent of big liver and spleen disease (BLS) and of hepatitis-splenomegaly syndrome (HSS) in chickens, which have caused economic losses to the poultry industry in China. In this study, 18 samples of BLS chickens were collected to reveal the molecular epidemiological characteristics of avian HEV in the province of Shandong, China. RESULTS: Gross and microscopic lesions of clinical samples were observed; then, virology detection and genetic analysis of avian HEV were performed. The results showed that there was significant swelling and rupture in the liver and that the spleen was enlarged. Microscopic lesions demonstrated obvious hemorrhage in the liver, with infiltration of heterophilic granulocytes, lymphocytes, and macrophages, as well as the reduction of lymphocytes in the spleen. Eleven of the 18 samples were positive for avian HEV, with a positive rate of 61.11%. More importantly, all avian HEV-positive samples were mixed infections: among these, the mixed infections of avian HEV and chicken infectious anemia virus (CIAV) and avian HEV and fowl adenovirus (FAdV) were the most common. Furthermore, the genetic evolution analysis showed that all avian HEV strains obtained here did not belong to the reported 4 genotypes, thus constituting a potential novel genotype. CONCLUSIONS: These results of this study further enrich the epidemiological data on avian HEV in Shandong, prove the genetic diversity of avian HEV in China, and uncover the complex mixed infections of avian HEV clinical samples.

First report of equine parvovirus-hepatitis and equine hepacivirus coinfection in horses in Korea.

Equine parvovirus-hepatitis (EqPV-H) and equine hepacivirus (EqHV) are etiologically associated with Theiler's disease (TD), causing fulminant equine hepatitis, but the transmission route and co-infection effect remain unclear. We determined EqPV-H and EqHV prevalence and coinfection rate in 160 serum and 114 faecal samples using nested polymerase chain reaction. Amino acid and nucleotide analyses were performed and phylogenetic trees were constructed. By measuring liver-specific parameters (AST, GGT, TBIL and A/G ratio), hepatopathological changes in viremia status were compared. We found a high prevalence (EqPV-H: 10.6% in serum, 5.3% in faeces; EqHV: 8.1% in serum) and coinfection rate (35.3% in EqPV-H) of TD-causing agents. The newly identified EqPV-H genomes showed high nucleotide and amino acid similarities with previously reported strains in the USA, China and Austria. In phylogenetic tree and recombination analysis, a natural recombination event was confirmed between Chinese and Korean strains. In the EqPV-H or EqHV viremic horses, AST was significantly elevated and at least two liver-specific parameters were outside the reference intervals in 43.5% (10/23) of horses. To our knowledge, this is the first prevalence field study of EqPV-H and EqHV using both serum and faeces, providing further evidence of faecal-oral transmission of TD. These epidemiologic and clinicopathologic analyses specify the risk factors of TD infection and promote disease prevention strategy.

A 10-Year Retrospective Study of Inclusion Body Hepatitis in Meat-Type Chickens in Spain (2011-2021).

A surge in fowl adenovirus (FAdV) causing inclusion body hepatitis (IBH) outbreaks has occurred in several countries in the last two decades. In Spain, a sharp increase in case numbers in broilers and broiler breeder pullets arose since 2011, which prompted the vaccination of breeders in some regions. Our retrospective study of IBH cases in Spain from 2011 to 2021 revealed that most cases were reported in broilers (92.21%) and were caused by serotypes FAdV-8b and -11, while cases in broiler breeder pullets were caused by serotypes FAdV-2, -11, and -8b. Vertical transmission was the main route of infection, although horizontal transmission likely happened in some broiler cases. Despite the inconsistent and heterogeneous use of vaccines among regions and over time, the number of cases mirrored the use of vaccines in the country. While IBH outbreaks were recorded year-long, significantly more cases occurred during the cooler and rainier months. The geographic distribution suggested a widespread incidence of IBH and revealed the importance of a highly integrated system. Our findings contribute to a better understanding of FAdV infection dynamics under field conditions and reiterate the importance of surveillance, serological monitoring of breeders, and vaccination of breeders against circulating serotypes to protect progenies.

Genomic Epidemiology and Evolution of Duck Hepatitis A Virus.

Duck hepatitis A virus (DHAV), an avian picornavirus, causes high-mortality acute disease in ducklings. Among the three serotypes, DHAV-1 is globally distributed, whereas DHAV-2 and DHAV-3 serotypes are chiefly restricted to Southeast Asia. In this study, we analyzed the genomic evolution of DHAV-1 strains using extant GenBank records and genomic sequences of 10 DHAV-1 strains originating from a large disease outbreak in 2004-2005, in Hungary. Recombination analysis revealed intragenotype recombination within DHAV-1 as well as intergenotype recombination events involving DHAV-1 and DHAV-3 strains. The intergenotype recombination occurred in the VP0 region. Diversifying selection seems to act at sites of certain genomic regions. Calculations estimated slightly lower rates of evolution of DHAV-1 (mean rates for individual protein coding regions, 5.6286 × 10-4 to 1.1147 × 10-3 substitutions per site per year) compared to other picornaviruses. The observed evolutionary mechanisms indicate that whole-genome-based analysis of DHAV strains is needed to better understand the emergence of novel strains and their geographical dispersal.

Equine Parvovirus-Hepatitis Screening in Horses and Donkeys with Histopathologic Liver Abnormalities.

There is strong evidence that equine parvovirus-hepatitis (EqPV-H) is associated with the onset of Theiler's disease, an acute hepatic necrosis, in horses. However, the impact of this virus on other hepatopathies remains unknown. The objective of this retrospective study was to evaluate the prevalence and quantify the viral loads of EqPV-H in formalin-fixed, paraffin-embedded equine and donkey livers with various histopathologic abnormalities. The pathologies included cirrhosis, circulatory disorders of the liver, toxic and metabolic hepatic diseases as well as neoplastic and inflammatory diseases (n = 84). Eight normal liver samples were included for comparison as controls. EqPV-H DNA was qualitatively and quantitatively measured by real-time PCR and digital PCR, respectively. The virus was detected in two livers originating from horses diagnosed with abdominal neoplasia and liver metastasis (loads of 5 × 103 and 9.5 × 103 genome equivalents per million cells). The amount of viral nucleic acids measured indicates chronic infection or persistence of EqPV-H, which might have been facilitated by the neoplastic disease. In summary, this study did not provide evidence for EqPV-H being involved in hepatopathies other than Theiler's disease.

Dual Circulation of Duck Hepatitis A Virus Genotypes 1 and 3 in Egypt.

Duck hepatitis A virus (DHAV) causes acute hepatitis and mortality, resulting in high economic losses in the duck farm industry. The current study describes the outbreak of DHAV in vaccinated duck farms in North Egypt during 2019 and molecular characterization of the 3' untranslated region (UTR) and viral protein VP1 genes. The 30 samples were collected from 7- to 28-day-old commercial Pekin ducks that showed a history of nervous signs and sudden deaths and were on farms in 6 governorates. DHAV was typed by reverse transcription-polymerase chain reaction (RT-PCR) for 3' UTR and VP1 genes and revealed 20 positive farms, with the first detection of DHAV genotype 3 (DHAV-3) in 18 samples and the classic DHAV-1 in 2 samples. The phylogenetic analysis of VP1 and 3' UTR genes of the nine selected strains representative of six governorates revealed that seven strains were clustered with DHAV-3 Chinese and Korean-Vietnamese strains within different subgroups with 92.4%-93.7% amino acid identity; such strains were distinguishable from the vaccine strain of DHAV-1 used in Egypt with 74.4% amino acid identity. The other strains were closely related to the DHAV-1 Asian strain and the vaccine strain used in Egypt with 98.7%-99.6% amino acid identity for the VP1 gene with different clustering than that of recently isolated DHAV-1 Egyptian strains. The VP1 gene of DHAV-3 had 1 hypervariable region (HVR) with 10 amino acid mutations compared with DHAV3/DN2/Vietnam/2011, but DHAV-1 had 3 HVRs with 1 amino acid mutation in HVRII compared with the DHAV-1 vaccine strain. In conclusion, a new introduction of DHAV-3 with the classical DHAV-1 was recorded in Pekin duck farms in North Egypt that is genetically distant from the vaccinal strain.

Artículo regular­Circulacíon dual de los genotipos 1 y 3 del virus de la hepatitis A del pato en Egipto. El virus de la hepatitis A del pato (con las siglas en inglés DHAV) causa hepatitis aguda y mortalidad, lo que genera grandes pérdidas económicas en la industria de la críanza de patos. El estudio actual describe un brote del virus de la hepatitis A del pato en una granja de patos vacunados en el norte de Egipto durante el año 2019 y la caracterización molecular de los genes de la región no traducida 3' (3' UTR) y la proteína viral VP1. Las 30 muestras se recolectaron de patos Pekin comerciales de 7 a 28 días de edad que presentaban antecedentes de signos nerviosos y muerte súbita y se encontraban en granjas de seis gobernaciones. El virus de la hepatitis A del pato se tipificó mediante la transcripción inversa y reacción en cadena de la polimerasa (RT-PCR) para los genes 3' UTR y VP1 y reveló 20 granjas positivas, con la primera detección del genotipo 3 del virus de la hepatitis A del pato (DHAV-3) en 18 muestras y la detección del virus clásico de la hepatitis A del pato tipo1 en dos muestras. El análisis filogenético de los genes VP1 y 3' UTR de las nueve cepas seleccionadas representativas de seis provincias reveló que siete cepas se agruparon con cepas del virus de la hepatitis A del pato 3 chinas y coreano-vietnamitas dentro de diferentes subgrupos con una identidad de aminoácidos del 92.4% al 93.7%; dichas cepas se distinguían de la cepa vacunal del virus de la hepatitis A del pato tipo 1 utilizada en Egipto con 74.4% de identidad de aminoácidos. Las otras cepas estaban estrechamente relacionadas con la cepa asiática del virus de la hepatitis A del pato tipo 1 y la cepa de vacuna utilizada en Egipto con 98.7% -99.6% de identidad de aminoácidos para el gene VP1 con agrupaciones diferentes a las de las cepas egipcias de virus de la hepatitis A del pato tipo 1 aisladas recientemente. El gene VP1 del virus de la hepatitis A del pato tipo 3 tenía una región hipervariable (HVR) con 10 mutaciones en la secuencia de aminoácidos en comparación con la cepa DHAV3/ DN2/Vietnam/2011, pero el virus de la hepatitis A del pato tipo 1 tenía tres regiones hipervariables con una mutación de aminoácidos en la zona hipervariable II en comparación con la cepa de vacuna virus de la hepatitis A del pato tipo 1. En conclusión, se registró una nueva introducción del virus de la hepatitis A del pato tipo 3 con el virus de la hepatitis A del pato clásico tipo 1 en granjas de patos Pekín en el norte de Egipto, que está genéticamente distante de la cepa vacunal.

Establishment of Duplex SYBR Green I-based Real-time PCR Assay for Simultaneous Detection of Duck Hepatitis A Virus-1 and Duck Astrovirus-3.

Duck viral hepatitis (DVH) mainly affects ducklings under 1 month of age, causes liver necrosis, enlargement, and hemorrhage, and is highly lethal, seriously jeopardizing the duck industry. The prevalence of duck hepatitis A virus (DHAV-1) and duck astrovirus type 3 (DAstV-3) is increasing, and coinfection is common. Moreover, the similar clinical characteristics of the DHAV-1 and DAstV-3 infections and the high frequency of coinfection make diagnosis difficult. In this study, to establish a method for the rapid, simultaneous detection of DHAV-1 and DAstV-3, two pairs of specific primers were designed according to their conserved gene regions. An SYBR® Green I-based qPCR assay was successfully established that can quickly and differentially detect the two viruses. Moreover, the assay is highly specific and does not show cross-reaction with other common viruses. The detection limit of the method is 7.34 × 101 copies/µl and 3.78 × 101 copies/µl for DHAV-1 and DAstV-3, respectively, indicating high sensitivity. A total of 34 clinical samples were tested using the established method; the positive rates for DHAV-1 and DAstV-3 were 14.71% and 8.82%, respectively, and that for coinfection was 2.94% (1/34), which was better than that obtained with conventional PCR. In summary, the SYBR Green I-based qPCR assay established in this study has high specificity, good sensitivity and accuracy, high feasibility, and is rapid. Thus, it can be a powerful tool for the coinfection detection of DHAV-1 and DAstV-3 and for future epidemiologic studies.

Artículo regular­Establecimiento de un ensayo dúplex de PCR en tiempo real basado en SYBR Green I para la detección simultánea del virus de la hepatitis A del pato-1 y del astrovirus del pato tipo 3. La hepatitis viral del pato (DVH) afecta principalmente a los patitos menores de 1 mes de edad, causa necrosis hepática, agrandamiento y hemorragia, y es altamente letal, lo que pone en grave peligro la industria del pato. La prevalencia del virus de la hepatitis A del pato (DHAV-1) y del astrovirus del pato tipo 3 (DAstV-3) está aumentando y la coinfección es común. Además, las características clínicas similares de las infecciones por el virus de la hepatitis A del pato y el astrovirus del pato tipo 3 así como la alta frecuencia de coinfección dificultan el diagnóstico. En este estudio, para establecer un método para la detección rápida y simultánea por el virus de la hepatitis A del pato y el astrovirus del pato tipo 3, se diseñaron dos pares de iniciadores específicos según sus regiones génicas conservadas. Se estableció con éxito un ensayo cuantitativo de PCR basado en SYBR® Green I que pudo detectar rápida y diferencialmente los dos virus. Además, el ensayo es muy específico y no muestró reacción cruzada con otros virus comunes. El límite de detección del método fue de 7.34 × 101 copias/µl y de 3.78 × 101 copias/µl para el virus de la hepatitis A del pato y para el astrovirus del pato tipo 3, respectivamente, lo que indica una alta sensibilidad. Se analizaron un total de 34 muestras clínicas utilizando el método establecido; las tasas positivas para el virus de la hepatitis A del pato y para el astrovirus del pato tipo 3 fueron del 14.71% y 8.82%, respectivamente y la de coinfección fue del 2.94% (1/34), que fue mejor que la obtenida con el método de PCR convencional. En resumen, el ensayo cuantitativo de PCR basado en SYBR Green I establecido en este estudio tiene alta especificidad, buena sensibilidad y precisión, alta viabilidad y es rápido. Por lo tanto, puede ser una herramienta poderosa para la detección de coinfecciones con el virus de la hepatitis A del pato y astrovirus del pato tipo 3 y para futuros estudios epidemiológicos.

Identification of a Putative Novel Genotype of Avian Hepatitis E Virus from Apparently Healthy Chickens in Southwestern Nigeria.

Avian hepatitis E virus (aHEV) is the major etiological agent of hepatitis-splenomegaly syndrome (HSS), big liver and spleen disease (BLSD), and hepatic rupture hemorrhage syndrome (HRHS) in chickens. Infections with aHEV cause a significant decrease in egg production and increased mortality in chickens worldwide. However, studies on the prevalence of aHEV in Nigeria are scarce. In this study, serum (n = 88) and fecal samples (n = 110) obtained from apparently healthy layer chickens from three states in southwestern Nigeria were analyzed by nested reverse transcription-polymerase chain reaction (nRT-PCR) targeting the helicase and capsid gene for the presence of aHEV. Avian HEV was detected in 12.5% (n = 11/88) of serum samples and 9.1% (n = 10/110) of fecal samples tested. Phylogenetic analysis showed that five of the twelve identified aHEV sequences belonged to genotype 2. The remaining seven sequences were only distantly related to other known aHEV isolates. After amplification of the near-complete ORF2 fragment (1618 bp) and part of the ORF1 (582 bp) of isolate YF40_aHEV_NG phylogenetic analysis revealed a nucleotide sequence identity between 79.0 and 82.6% and 80.1 and 83.5%, respectively, to other known aHEV strains, indicating that the Nigerian isolate YF40_aHEV_NG belongs to a novel aHEV genotype. This is the first report of co-circulation of aHEV genotypes in chickens in Nigeria.

Genome characterization, prevalence and tissue distribution of astrovirus, hepevirus and norovirus among wild and laboratory rats (Rattus norvegicus) and mice (Mus musculus) in Hungary.

Rodents including rats are reservoir of several pathogens capable of affecting human health. In this study, faecal and different organ specimens from free-living Norway rats (Rattus norvegicus) (N = 18) and faecal samples from laboratory rodents (rats N = 21 and mice N = 20) collected from different geographic areas in Hungary between 2017 and 2020 were investigated by viral metagenomics and conventional RT-PCR methods. The complete genome of three different RNA viruses, rat astrovirus, rat norovirus and rat hepevirus were characterized and analysed in detail. Rat norovirus was detected in faecal (17.6%, 3/17) and kidney (7.1%, 1/14) samples; rat astrovirus in faecal (23.5%, 4/17) and spleen (13.3%, 2/15) samples, and rat hepevirus in 43% to 67% the faecal, liver, kidney, lung, heart, muscle, brain and blood samples from Norway rats, respectively. Rat norovirus was also identifiable in 5% (1/21) of laboratory rats and rat astrovirus in 40% (8/20) of faecal samples from laboratory mice. Co-infections were found in 28% (5/18) wild Norway rats. The highest RNA viral load of astrovirus (1.81 × 108 copy/g) and norovirus (3.49 × 107 copy/g) were measured in faecal samples; while the highest RNA viral load of hepevirus (1.16 × 109 copy/g) was found in liver samples of Norway rats, respectively. This study confirms the wide geographic distribution and high prevalence of astrovirus, norovirus and hepevirus among wild rats in Hungary with confirmation of different organ involvement of as well as the detection of norovirus and astrovirus in laboratory rats and mice, respectively. This finding further strengthens the role of rodents in the spread of viral pathogens especially infecting human.

Serological evidence of avian HEV antibodies in apparently healthy chickens in southwest Nigeria.

Avian hepatitis E virus (aHEV) is associated with hepatitis-splenomegaly syndrome, big liver and spleen disease and hepatic rupture haemorrhage syndrome. However, the knowledge about aHEV in commercial layer chickens in Nigeria is scarce. In this study, 460 serum samples obtained from 36 apparently healthy commercial layer chicken flocks in three states (Ogun, Osun and Oyo States) of southwestern Nigeria were analysed by enzyme linked immunosorbent assay for the presence of anti-aHEV immunoglobulin Y (IgY) antibodies. In total, the overall seroprevalence of anti-aHEV antibodies was 14.6%. The serological analysis revealed that 75% of the flocks examined were positive for anti-aHEV IgY antibodies from chickens of various ages in all three states. The percentage of the seropositive chickens in the three states varied from flock to flock ranging from 60% to 88.8% and seropositive chickens were detected at any age (24-52 weeks of age) without significant differences between the age groups. This is the first report assessing the presence of aHEV antibodies in chickens from Nigeria. The detection of anti-aHEV antibodies in commercial layer chickens in this study emphasizes the importance of serosurveillance in disease monitoring due to the economic threat posed by aHEV as a result of decreased egg production and increased mortality in affected commercial layer chicken farms. However, further studies are essential to reveal the clinical implications and to assess the real burden of aHEV in Nigeria.

Serologic and Molecular Evidence of Widespread Infection of Avian Hepatitis E Virus in Poultry Farms of Iran.

Hepatitis-splenomegaly syndrome is caused by avian hepatitis E virus (aHEV), a nonenveloped, single-stranded RNA virus. The economic importance of this disease in the poultry industry is due to the decline in egg production (10%-40%) and the rise in mortality (1%-4%). In the present study, 1540 serum samples from 33 broiler breeder flocks were analyzed by an enzyme-linked immunosorbent assay for the presence of an anti-aHEV antibody. In addition, a diagnostic nested reverse transcriptase-PCR was done on all farm samples. In the serologic study, 66.7% (22/33) of the flocks and 28.5% (439/1540) of the chickens were positive. The molecular study showed that three farms were positive, and PCR products were observed for the conserved regions of the aHEV helicase and capsid virus genes as 386 bp and 242 bp, respectively. It should be noted that clinical and pathologic symptoms including decreased egg production, enlarged livers and spleens, and a slight rise in mortality rate were observed in eight farms. To our knowledge, this is the first documented study on the aHEV identification and its antibody detection in broiler breeder farms in Iran.

Evidencia serológica y molecular de una infección diseminada del virus de la hepatitis E aviar en granjas avícolas en Irán. El síndrome de hepatitis-esplenomegalia es causado por el virus de la hepatitis E aviar (aHEV), un virus de ARN de cadena simple sin envoltura. La importancia económica de esta enfermedad en la industria avícola se debe a la disminución en la producción de huevo (10%-40%) y al aumento de la mortalidad (1%-4%). En el presente estudio, se analizaron 1540 muestras de suero de 33 parvadas de reproductores pesados mediante un ensayo de immunoabsorción con enzimas ligadas para determinar la presencia de anticuerpos contra el virus de la hepatitis E aviar. Además, se realizó un método de transcripción reversa y PCR anidado de diagnóstico en todas las muestras de la granja. En el estudio serológico, el 66.7% (22/33) de las parvadas y el 28.5% (439/1540) de los pollos fueron positivos. El estudio molecular mostró que tres granjas fueron positivas, y se observaron productos de PCR para las regiones conservadas de los genes del virus de la cápside y de la helicasa del virus de la hepatitis E aviar con tamaños de 386 pb y 242 pares de bases, respectivamente. Cabe señalar que en ocho granjas se observaron signos clínicos y patológicos como disminución de la producción de huevos, agrandamiento del hígado y del bazo y un ligero aumento en la tasa de mortalidad. Hasta donde se conoce, este es el primer estudio documentado sobre la identificación del virus de la hepatitis E aviar y la detección de anticuerpos en granjas de pollos de engorde en Irán.

Genetic diversity of avian hepatitis E virus in China, 2018-2019.

Avian hepatitis E virus (HEV) is highly variable and has multiple genotypes. Previous studies showed that the current epidemic strain in China is genotype 5, but the relevant detection was only carried out in flocks with hepatic rupture haemorrhage syndrome, which does not mean that other genotypes do not exist. In this study, a broader analysis involving different chicken flocks was performed to understand the epidemic status of avian HEV in China. The results showed that the HEV-positive rate of all samples was 7.92% (78/985), and four different genotypes have been identified by analysing a truncate capsid gene fragment, while the homology between them is about 80%. Two of them are separately known as genotype 3 and genotype 5, while the other two are completely unidentified, indicating that there are multiple genotypes of avian HEV prevalent in China. At the same time, the distribution of these genotypes has no obvious geographical clustering pattern, only slightly different in commercial layers, broilers and some indigenous species. This study shows the genetic diversity of avian HEV in China and reminds us to pay more attention to its variation and evolution.

Co-infection with avian hepatitis E virus and avian leukosis virus subgroup J as the cause of an outbreak of hepatitis and liver hemorrhagic syndromes in a brown layer chicken flock in China.

Hens of a commercial Hy-line brown layer flock in China exhibited increased mortality and decreased egg production at 47 wk of age. From 47 to 57 wk, average weekly mortality increased from 0.11 to 3.0%, and egg production decreased from 10 to 30%, with a peak mortality rate (3.0%) observed at 54 wk of age. Necropsy of 11 birds demonstrated tissue damage that included hepatitis, liver hemorrhage, rupture, and/or enlarged livers. Microscopic liver lesions exhibited hepatocytic necrosis, lymphocytic periphlebitis, and myeloid leukosis. While no bacteria were recovered from liver and spleen samples, avian hepatitis E virus (HEV) RNA was detected in all 11 tested hens by nested reverse transcription-polymerase chain reaction. Of these, subgroup J avian leukosis virus (ALV-J) proviral DNA was detected in 5 hens by PCR. Alignments of partial ORF2 gene sequences obtained here demonstrated shared identity (76 to 97%) with corresponding sequences of other known avian HEV isolates. Env sequences of ALV-J isolates obtained here shared 50.1 to 55% identity with other ALV subgroups and 91.8 to 95.5% identity with other known ALV-J isolates. Phylogenetic tree analysis of selected sequences obtained here grouped an avian HEV sequence with genotype 3 HEV and assigned an ALV-J sequence to a branch separate from known ALV-J subgroups. Immunohistochemical results confirmed the presence of avian HEV and ALV-J in livers. Therefore, these results suggest that avian HEV and ALV-J co-infection caused the outbreak of hepatitis and liver hemorrhagic syndrome observed in the layer hen flock analyzed in this study.

Characterization of Equine Parvovirus in Thoroughbred Breeding Horses from Germany.

An equine parvovirus-hepatitis (EqPV-H) has been recently identified in association with equine serum hepatitis, also known as Theiler's disease. The disease was first described by Arnold Theiler in 1918 and is often observed with parenteral use of blood products in equines. However, natural ways of viral circulation and potential risk factors for transmission still remain unknown. In this study, we investigated the occurrence of EqPV-H infections in Thoroughbred horses in northern and western Germany and aimed to identify potential risk factors associated with viral infections. A total of 392 Thoroughbreds broodmares and stallions were evaluated cross-sectionally for the presence of anti-EqPV-H antibodies and EqPV-H DNA using a luciferase immunoprecipitation assay (LIPS) and a quantitative PCR, respectively. In addition, data regarding age, stud farm, breeding history, and international transportation history of each horse were collected and analysed. An occurrence of 7% EqPV-H DNA positive and 35% seropositive horses was observed in this study cohort. The systematic analysis of risk factors revealed that age, especially in the group of 11-15-year-old horses, and breeding history were potential risk factors that can influence the rate of EqPV-H infections. Subsequent phylogenetic analysis showed a high similarity on nucleotide level within the sequenced Thoroughbred samples. In conclusion, this study demonstrates circulating EqPV-H infections in Thoroughbred horses from central Europe and revealed age and breeding history as risk factors for EqPV-H infections.