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Tembusu virus (TMUV) belongs to the genusFlavivirusthat may cause severe egg drop in ducks. In order to evaluate the most efficient TMUV detection method, the performances of a conventional RT-PCR (C-RT-PCR), a semi-nested PCR (SN-RT-PCR), a reverse-transcriptase real-time quantitative PCR (Q-RT-PCR), and a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) targeting the TMUV virus-specific NS5 gene were examined. In order to compare the sensitivity of these four techniques, two templates were used: (1) plasmid DNA that contained a partial region of the NS5 gene and (2) genomic RNA from TMUV-positive cell culture supernatants. The sensitivities using plasmid DNA detection by C-RT-PCR, SN-RT-PCR, Q-RT-PCR, and RT-LAMP were 2 × 104copies/μL, 20 copies/μL, 2 copies/μL, and 20 copies/μL, respectively. The sensitivities using genomic RNA for the C-RT-PCR, SN-RT-PCR, Q-RT-PCR, and RT-LAMP were 100 pg/tube, 100, 10, and 100 fg/tube, respectively. All evaluated assays were specific for TMUV detection. The TMUV-specific RNA was detected in cloacal swabs from experimentally infected ducks using these four methods with different rates (52–92%), but not in the control (non-inoculated) samples. The sensitivities of RT-PCR, SN-RT-PCR, Q-RT-PCR, and RT-LAMP performed with cloacal swabs collected from suspected TMUV infected ducks within 2 weeks of severe egg-drop were 38/69 (55.1%), 52/69 (75.4%), 57/69 (82.6%), and 55/69 (79.7%), respectively. In conclusion, both RT-LAMP and Q-RT-PCR can provide a rapid diagnosis of TMUV infection, but RT-LAMP is more useful in TMUV field situations or poorly equipped laboratories. [ABSTRACT FROM AUTHOR]

Outbreaks of fowl adenoviruses (FAdVs) infection with the main clinical signs of hepatitis-hydropericardium-syndrome or inclusion body hepatitis have been frequently reported in ducks and geese in recent years, causing economic losses for the Chinese waterfowl industry. This study investigated 792 samples (391 fattening ducks, 192 breeder ducks and 209 fattening geese) of suspected FAdV-infected waterfowl from 38 farms (21 fattening duck farms, nine breeder duck farms and eight fattening geese farms) in Shandong Province between 2019 and 2022. The results showed a 60.23% infection rate for FAdVs (477/792), while the infection rate for breeder ducks was almost the same as that for fattening geese (55.73% vs. 54.55%). Notably, co-infection with avian influenza virus H9N2 (H9N2), Tembusu virus (TMUV), duck hepatitis virus (DHV), duck circovirus (DuCV), goose astrovirus (GAstV) and duck parvovirus (DPV)/goose parvovirus (GPV) was common among the 477 FAdVs positive cases. Phylogenetic analysis of complete hexon genes of 22 FAdV strains in Shandong Province showed the presence of four species (FAdV-A, C, D, E) and five serotypes (FAdV-1, 4, 8a, 8b, 11). These findings provide the first data on the prevalence and co-infection status of FAdVs in Shandong Province and can be used as a basis for FAdV prevention in the field. Samples of suspected FAdV-infected waterfowl from farms in Shandong Province were collected from 2019 to 2022. Single infections with FAdV were less frequent than mixed infections. 477 out of 792 samples (60.23%) tested positive for FAdV nucleic acids. Detection rate of FAdV was 65.47% in fattening duck farms, 55.73% in breeder duck farms and 54.55% in fattening geese farms. [ABSTRACT FROM AUTHOR]

Poultry production in China has been experiencing a high incidence of broiler arthritis /tenosynovitis caused by avian orthoreovirus (ARV) since 2013. In the spring of 2020 severe arthritis cases from broiler flocks were identified in a large-scale commercial poultry company in Anhui Province, China. Diseased organs from dead birds were sent for diagnosis to our laboratory. ARVs, including seven broiler-isolates and two breeder-isolates, were successfully harvested and sequenced. Interestingly, the genotypes of ARVs isolated from infected chickens were inconsistent between different flocks, or even between different houses on the same flocks. Pathogenicity testing in chicks confirmed that the seven broiler-isolates were pathogenic strains, which could cause arthritis in infected chickens. Subsequently, a total of 89.66% serum samples collected from apparently healthy adult broiler flocks not vaccinated against ARV tested positive for ARV antibodies, suggesting that low and high virulence reovirus strains may be co-circulating in the farm. To this end, we collected dead embryos of unhatched chicken eggs for pathogen tracing, and the two ARV breeder-isolates isolated indicated that vertical transmission from breeders to progeny should not be underestimated for the prevalence of ARV within broiler flocks. The findings have implications for the evidenced-based formulation of prevention and control strategies. [ABSTRACT FROM AUTHOR]

Avian orthoreovirus (ARV) infections of broiler flocks cause arthritis/tenosynovitis syndrome and significant economic losses. ARV variants were detected in the USA and Canada. Viral arthritis/tenosynovitis syndrome has occurred frequently in China in recent years. In this study, a variant ARV strain associated with viral arthritis/tenosynovitis syndrome was isolated from broilers and designated as LY383. Genomic sequence and phylogenetic analysis of the σC nucleic acid and amino acid sequences revealed that the isolate was closely related to ARV field strains Reo/PA/Layer/01224B/14, Reo/PA/Broiler/1551/13, GA/14602/2014, GA/13569/2013 and GA/13542/2013, in cluster V, but distinct from most Chinese field strains or commercial vaccine strains. Experimental challenge showed that the isolate could cause arthritis/tenosynovitis syndrome in broilers, which possessed a high level of maternal antibodies induced by commercial ARV vaccines (S1133, 1733 and T98). Furthermore, viral nucleic acid could be detected in cloacal swabs of all challenged birds throughout the entire test from 5 dpi onward. These results suggest that a novel ARV genotype emerges and might become prevalent in broiler flocks in China. RESEARCH HIGHLIGHTS A variant avian orthoreovirus was isolated from a vaccinated broiler flock in North China. The ARV field strain was distinct from previous China-origin ARV isolates and vaccine strains. The current commercial ARV vaccine could not provide effective protection of broilers against the field isolate infection. These findings indicated that variant ARV field strains might become frequent in broiler flocks in China and effective measures should be conducted to prevent and control the disease. [ABSTRACT FROM AUTHOR]

Cherry Valley breeder ducks in Shandong province in northern China have experienced swollen eyes, lachrymation, pox scabs on glabrous or glabrous skin, depression and dysentery since 2021. The spread of this infectious disease has seriously affected the breeder ducks in major Cherry Valley duck farms in China. The virus causing clinical signs in Cherry Valley breeder ducks was isolated by chicken embryo inoculation. We successfully isolated a strain of duck pox virus from diseased breeder ducks by virus replication. We have also successfully conducted experiments for duck pox disease using the isolated strains to infect ducklings. By comparison with 22 pox viruses already published in GenBank, the virus strain obtained in this study was most homologous (about 99.7%) to the strain isolated from infected domestic ducks in Guangxi, China in 2014 (KJ192189), and belonged to the same A5 subtype. Since there were no previous cases of avian pox virus infecting white or Cherry Valley duck breeds, this study identified a new host for avian pox virus infection and provided theoretical support and data for the development of avian pox virus research. RESEARCH HIGHLIGHTS Avian pox virus can infect a new host type – Cherry Valley breeder ducks. The avian pox virus isolated from Cherry Valley breeder ducks showed highest identity with Guangxi hemp duck-derived avian pox virus. Cherry Valley breeder ducks were infected with avian pox virus of subtype A5. [ABSTRACT FROM AUTHOR]

Hydropericardium hepatitis syndrome (HHS) is a fatal disease in chickens, mainly caused by fowl adenovirus serotype 4 (FAdV-4). Since June 2015, HHS has appeared in many provinces in China. The disease has spread from broilers to laying hens, breeders and Cherry Valley ducks, seriously endangering the health of the poultry industry in China. In July 2016, an infectious disease was noticed in a goose farm in Jinan, Shandong Province, China, and hydropericardium was the main finding in post mortem investigations. In the actual study, we isolated a FAdV-4 strain from the livers of naturally-infected goslings and designated it as SDJN. We first evaluated its pathogenicity by inoculating Taizhou geese at 10, 20, and 30 days of age with 10−7.15EID50/0.2 ml doses of the SDJN strain in 1 ml allantoic fluid via subcutaneous injection or oral infection. Clinical signs and pericardial effusion appeared in geese infected subcutaneously at 10 days of age, whereas 20- and 30-day-old geese were not susceptible to FAdV-4. The results of real-time PCR showed that the replication ability of FAdV-4 in geese correlated with the age. Furthermore, results from clinical chemistry showed that FAdV-4 damaged the liver and kidney in geese and the results paralleled viral load and gross lesions. Consequently, FAdV-4 was pathogenic in geese, and the pathogenicity was related to age and mode of infection. This study is the first experimental infection of FAdV-4 in geese, which will provide a basis for further understanding of the disease. RESEARCH HIGHLIGHTS Pathogenicity tests with a FAdV-4 were conducted in geese, which included data on clinical signs, gross pathology, histopathology, clinical chemistry and viral load. FAdV-4 could replicate in geese and HHS was successfully induced. Pathogenicity of FAdV-4 in geese was related to the age and routes of infection. [ABSTRACT FROM AUTHOR]