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Avian hepadnaviruses are etiological agents of hepatitis B, that has been identified primarily in ducks, and more recently in various avian species. In this paper, 16 hepadnaviruses were detected by polymerase chain reaction (PCR) in the field samples from dead embryos of commercially reared domestic duck and goose. Based on the molecular analysis of the S-protein gene sequences and phylogenetic Neighbor-joining tree, identified viruses were clustered in the same genetic group, indicating no host-related diversity. Both duck and goose-origin hepadnaviruses were grouped within the cluster consisting of "Western-country" and "Chinese" duck hepatitis B (DHBV) isolates, showing more evolutionary distances with other known avian hepadnaviruses. Histopathologically, the lesions observed in the liver tissue from hepadnavirus positive duck and goose embryos varied from low to mild degree of perivascular mononuclear cells and mixed cell infiltrations, followed by mild vacuolar changes. Small focal necrotic changes in the liver parenchyma, and bile ductular proliferation were also found in examined liver samples. Generally, the microscopic findings resemble those described in experimentally infected ducks, while this was the first description of hepadnavirus associated lesions in domestic goose. Although hepadnaviruses are considered to have a very narrow host range, this study showed that domestic ducks and geese are susceptible to infection with genetically almost identical hepadnaviruses, that were likely to produce similar microscopic changes in the liver of both duck and goose embryos. The impact of naturally occurred hepadnavirus infection and possible synergistic interactions with other infectious or non-infectious agents on embryo viability needs further investigation., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The natural occurrence of chicken and turkey-origin astroviruses in domestic ducks (Anas platyrhynchos domesticus) is described. Twenty-two duck flocks were covered by this research. The liver, spleen, kidney and intestines were sampled and tested by reverse-transcription polymerase chain reaction for the presence of avian nephritis virus (ANV), chicken astrovirus (CAstV), turkey astrovirus (TAstV)-1, TAstV-2 and duck astrovirus. The astrovirus infection was confirmed in multiple organ samples from 59.1% of tested flocks. CAstV was detected in one flock, TAstV-2 in three flocks and ANV in 10 flocks. The molecular and phylogenetic analysis of the small open reading frame (ORF) 1b fragment (130 nucleotides) of all chicken and turkey-origin astroviruses detected in ducks showed that ANV-sequence group was more distant from CastV, TAstV-1 and TAstV-2 sequences, which formed a separate, more related group. ANV sequences were divided into three subgroups, suggesting that several types of ANV were circulating in Croatian duck flocks. The comparison of the partial ORF 1b (254 nucleotides) duck ANV sequences with 21 ANVs detected in various avian species (chickens, turkeys, geese, guinea fowl and pigeons) revealed they shared the higher nucleotide (95.6 to 97.2%) and amino acid (98.8 to 100%) identity with two ANV-2-like sequences from chickens (GA-SEP-A451-05 and GA-CK-SEP ANV-364-2005). Phylogenetic neighbour-joining tree analysis based on the same nucleotide alignment, and performed using the Jukes-Cantor method, clustered the compared sequences into three groups. All analysed duck ANV sequences showed a close phylogenetic relationship with chicken-origin ANVs. Additional work is required to determine the significance and pathogenicity of chicken and turkey-origin astroviruses in domestic ducks.

The first detection of avian nephritis virus (ANV) in goose embryos and of turkey astrovirus-1 (TAstV-1) in duck embryos is described. Intestinal samples from duck and goose embryos from five duck and four goose flocks in Croatia were tested by polymerase chain reaction for the presence of ANV, TAstV-1, turkey astrovirus-2, chicken astrovirus, duck astrovirus and also for the presence of avian reovirus, Derzsy's disease virus and duck enteritis virus. The kidneys from duck and goose embryos were also tested for ANV, while liver samples were tested for duck astrovirus. Duck embryos were also tested to detect duck circovirus and goose embryos for the presence of goose circovirus and goose haemorrhagic polyomavirus. All embryos were in the final stage of incubation and were characterized by moderate to markedly retarded growth. ANV was confirmed in the intestines and kidneys of embryos from two duck and two goose flocks and TAstV-1 was found in embryos from two duck flocks. One duck flock was positive for both ANV and TAstV-1. No other viruses were found in tested flocks. Phylogenetic analysis based on the ANV polymerase gene fragment of ANV sequences detected in duck and goose embryos revealed greatest similarity (88.1 to 97.2%) with ANV isolates from chickens. Further, the existence of at least two types of ANV circulating in Croatian duck and goose flocks was confirmed. Based on the phylogenetic analysis of the portion of TAstV-1 polymerase gene, two detected TAstV-1 nucleotide sequences were 99.5% similar. Compared with six TAstV-1 sequences, Croatian sequences showed one unique nucleotide change. In addition to other possible causes of stunted growth and late embryonic death, these findings suggest that ANV and/or TAstV-1 infection may be a contributing factor in the pre-hatching mortality of ducklings and goslings.

The first evidence of avian nephritis virus (ANV) in ducks is described. A diagnostic investigation was performed on three duck farms in Croatia. Samples from dead-in-shell ducklings and ducklings aged 30 days were collected and prepared for molecular and histopathological examination. Intestinal and liver samples were tested by polymerase chain reaction (PCR) for the presence of ANV, duck enteritis virus, duck hepatitis virus 1 and Derzsy's disease virus. Multiple tissues were collected for histological examination and lesions were found to be confined to the kidney and intestine. Moderate focal interstitial and periglomerular mononuclear cell infiltrates (mostly lymphocytes and plasma cells) were detected in the kidney. The duodenum showed rather diffuse pericryptal mononuclear cell hyperplasia (lymphocytes) and fibroplasia. ANV was detected by PCR in all the intestinal samples, while no other viruses were found. Sequence comparisons of the portion of the open reading frame 1b encoding the RNA-dependent RNA polymerase gene confirmed that the virus detected and sequenced from ducklings shared high nucleotide and amino acid identities with ANV-1. Additional work is required to determine the clinicopathological significance of ANV infection in ducks.