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Tembusu virus (TMUV) has been a causative agent of an acute egg-drop syndrome found in Chinese duck populations since at least 2010. In this paper, we report the characterization of a TMUV-like flavivirus (named CJD05) isolated from naturally infected egg-laying fowl. The virus was identified and then isolated from hens suffering from severe egg drop and fever in Fujian Province, China. The virus replicated well in MDEF and CEF cells, and its cytopathogenic effect (CPE) was apparent. Hemagglutinating activity (HA) was negative for this virus using erythrocytes from both chickens and pigeons. Viral particles were enveloped and approximately 45 nm in diameter, as observed by electron microscopy. Phylogenetic analysis of the full-length nucleotide sequence of CJD05 indicated that this virus is closely related to the duck-origin TMUV, belonging to Ntaya group of flavivirus. Most importantly, pathogenicity studies showed that CJD05 is highly virulent in 1-day-old chicks, 1-day-old Muscovy ducks, egg-laying chickens and shelducks. Our research highlights the increase in epidemic disease caused by avian TMUV, and subsequent outbreaks are becoming more complicated to treat. The pathogenic mechanisms of the virus are still not fully understood, further research is needed. [ABSTRACT FROM AUTHOR]

Muscovy duck-origin goose parvovirus (MDGPV) is a new virus resulting from the natural recombination of Muscovy duck parvovirus (MDPV) and goose parvovirus (GPV). Previously identified MDGPV strains were found to have two recombination regions, one in the P9 promoter to the NS region and one in the VP3 gene, or only one recombination in the VP3 gene. In 2022, a novel strain of MDGPV known as 2022FZ was identified from China's mainland. Complete genome sequence analysis showed that there were three recombination regions in this strain: one located in the P9 promoter-NS (425–612 nt) region, one in the NS2 (1,483–1,824 nt) region, and one in the VP3 (3,124–4,248 nt) region, respectively. The recombination regions in the P9 promoter-NS, NS2, and VP3 genes were substituted with the relevant GPVs sequences, whereas the MDPV virulent strain served as the skeleton in this instance. In addition, the 2022FZ strain had multiple unique aa mutations in the NS protein and the VP protein. The Muscovy duckling challenge test showed that MDGPV-2022FZ is less pathogenic to Muscovy ducklings than two recombinant or sole recombinant MDGPV strains. For the first time, our study identified a three-region recombinant MDGPV strain and detected the novel recombination event in the NS2 gene. These results contribute to our understanding of the pathogenicity and genetic diversity of duck parvovirus. [ABSTRACT FROM AUTHOR]

A multiplex polymerase chain reaction (PCR) method was developed to detect and distinguish goose parvovirus (GPV), waterfowl reovirus (WRV), and goose astrovirus (GAstV). Three pairs of primers were designed based on conserved regions in the genomic sequences of these enteric viruses and were used to specifically amplify targeted fragments of 493 bp from the viral protein 3 (VP3) gene of GPV, 300 bp from the sigma A-encoding gene of WRV, and 156 bp from the capsid protein-encoding gene of GAstV. The results showed that the primers can specifically amplify target fragments, without any cross-amplification with other viruses, indicating that the method had good specificity. A sensitivity test showed that the detection limit of the multiplex PCR method was 1 × 103 viral copies. A total of 102 field samples from Muscovy ducks with clinically suspected diseases were evaluated using the newly developed multiplex PCR method. The ratio of positive samples to total samples for GPV, WRV, and GAstV was 73.53% (75/102) for multiplex PCR and was 73.53% (75/102) for routine PCR. Seventy-five positive samples were detected by both methods, for a coincidence ratio of 100%. This multiplex PCR method can simultaneously detect GPV, WRV, and GAstV, which are associated with viral enteritis, thereby providing a specific, sensitive, efficient, and accurate new tool for clinical diagnosis and laboratory epidemiological investigations. • A multiplex PCR assay was developed for simultaneous detection for GPV, WRV, and GAstV in Muscovy duck flocks, with good specificity, good accuracy, and high sensitivity. • The multiplex PCR was able to detect at least 1 × 103 viral copies of GPV, WRV, and GAstV. • The multiplex PCR developed is a useful tool for clinical diagnosis of single virus and/or mixed infections in Muscovy duck. [ABSTRACT FROM AUTHOR]