Infectious bronchitis of chicken is a high morbidity and mortality viral disease affecting the poultry industry worldwide; therefore, a better understanding of this pathogen is of utmost importance. The primary aim of this study was to obtain a deeper insight into the genomic diversity of field infectious bronchitis virus (IBV) strains using phylogenetic and recombination analysis. We sequenced the genome of 20 randomly selected strains from seven European countries. After sequencing, we created a genome sequence data set that contained 36 European origin field isolates and 33 vaccine strains. When analyzing these 69 IBV genome sequences, we identified 215 recombination events highlighting that some strains had multiple recombination breaking points. Recombination hot spots were identified mostly in the regions coding for non-structural proteins, and multiple recombination hot spots were identified in the nsp2, nsp3, nsp8, and nsp12 coding regions. Recombination occurred among different IBV genotypes and involved both field and vaccine IBV strains. Ninety percent of field strains and nearly half of vaccine strains showed evidence of recombination. Despite the low number and the scattered geographical and temporal origin of whole-genome sequence data collected from European Gammacoronaviruses, this study underlines the importance of recombination as a major evolutionary mechanism of IBVs.
Fowl adenovirus D is the main cause of inclusion body hepatitis in chickens. Whole genome sequencing was carried out to enrich the genomic sequence database using field isolates of FAdV-D. Out of 44 newly determined genomes, 43 were classified into FAdV-2/-11 and 2 into FAdV-3; no FAdV-9 was identified. Whole-genome based phylogeny showed that FAdV-3 was more distantly related to FAdV 9 and FAdV-2/-11 than FAdV-9 and FAdV-2/-11 to each other. Whole-genome sequence homology analysis revealed that the full-length FAdV-3 genome harbored a ~12 kbp fragment of the genome that shared moderate sequence homology with representative strains of other FAdV-D serotypes but high relatedness with only the FAdV-3 strain whose full-genome is available in GenBank. A closer look onto the fiber and the penton genes of our FAdV-3 isolate identified putative recombination events; both the fiber and the penton coding genes shared fragments originating from FAdV-9. Of interest, ORF19 displayed a close relationship with the homologous genomic region of some FAdV-E strains (amino acid sequence homology, up to 82%). Thus, although FAdV-3 is classified into FAdV-D, the genomic structure of FAdV-3 appears to result from multiple heterotypic and heterologous recombination events. This study highlights the unique origin of FAdV-3.
Adenovirus (AdV) infection has been rarely documented in cats and other felids. Partial sequences of the hexon and fiber genes of a Hungarian feline adenovirus isolate (FeAdV isolate) showed a close relationship to human AdV (HAdV) type C1. Further molecular and biological characterization is reported here. Whole-genome sequencing revealed two silent mutations in the genome of the FeAdV isolate compared to a HAdV-C1 reference strain (at positions 14,096 and 15,082). Competitive antibody binding to the Coxsackie–adenovirus receptor and αvβ3 and αvβ5 integrin coreceptors inhibited the binding of the FeAdV isolate in different cell lines, but residual infections suggested alternative entry routes. The FeAdV isolate was found to be more sensitive to heat, low pH and detergents, but more resistant to alkaline and free chlorine treatments, as well as to ribavirin, stavudine and cidofovir treatments, than other human AdV types. We observed a suppression of IL-10 and TGF-β1 production during the entire course of viral replication. This immunomodulation may restore intratumoral immunity; thus, the FeAdV isolate could serve as an alternative oncolytic vector. Collectively, our results support that the Hungarian FeAdV isolate is a variant of common HAdV-C1. The cohabitation of cats with humans might result in reverse zoonotic infection. Felids appear to be susceptible to persistent and productive adenovirus infection, but further studies are needed to better understand the clinical and epidemiological implications.
Although the clinical course and pathogenesis of lymphocystis disease virus (LCDV) infection have been extensively described in freshwater and seawater environments, lymphocystis disease has not been studied in the copperband butterflyfish (Chelmon rostratus) or described at the molecular level in orbicular batfish (Platax orbicularis). The present study aimed to identify LCDV in a copperband butterflyfish and an orbicular batfish using light and electron microscopy (morphological) and molecular methods, namely PCR followed by phylogenetic analysis. We present a case series of two representatives of two distinct fish species with stress-induced chronic LCDV infection, which presented with typical, recurring, macroscopically visible lymphocystis nodules on their pectoral, caudal, and dorsal fins. After collecting lymphocystis nodules from live animals using skin scraping, we processed the hypertrophic giant cells for qualitative analysis using light and electron microscopy. Through our qualitative morphological analysis, we also share intimate observations of putative viral replication and assembly in the intracytoplasmic inclusion bodies of lymphocystis nodules. We present LCDV infection in a novel species, the copperband butterflyfish, and our molecular analysis identified the virus from the orbicular batfish as a novel LCDV species.
Avian pox is a globally spread viral disease affecting a wide spectrum of wild and domesticated bird species. The disease is caused by a diverse group of large DNA viruses, namely, avipoxviruses (genus Avipoxvirus, family Poxviridae). In this study, gross pathological examination and histopathological examination of skin lesions and several organs suggested acute poxvirus infection of a Eurasian crane (Grus grus, Linnaeus, 1758). Avipoxvirus infection was confirmed by testing wart-like lesions via gene-specific PCR assay and sequencing the obtained amplicon. Phylogenetic analysis of the gene encoding the DNA polymerase revealed that the crane poxvirus clustered in clade A, subclade A3. A large fragment of the poxvirus genome (306,477 bp in length) was assembled from the DNA of a skin specimen. Our study reaffirms previous findings that even complex virus genomes can be determined from a metagenomic assemblage generated directly from avian tissue samples without prior virus isolation, a promising approach for the epidemiologic surveillance of avipoxvirus infections in wild birds and domestic poultry.