The aim of the study was to determine the effect of field strain, serotype 7 (FAdV-7 JN-5/10), of fowl adenovirus infection on the replication of Rispens/CVI988 strain of Marek’s disease virus. Ninety one-day-old SPF chickens were divided into six groups. The chickens from group I were vaccinated against Marek’s disease (MD) and 24 h later infected with the adenovirus; chickens from group II were vaccinated and infected simultaneously; chickens from group III were infected with the adenovirus and 24 h after the infection vaccinated against MD. The chickens from groups IV-VI were: control of infection (IV), control of vaccination (V), and neither vaccinated nor infected (VI). After 3, 7, 14, 21, and 28 d post infection, the number of copies of pp38 gene of Rispens strain and hexon gene of FAdV strain was determined in the bursa of Fabricius and liver using real-time PCR. The results indicated that in all cases the replication of Rispens strain was reduced to about 1.0 log10 - 3.5 log10 in chickens infected with the adenovirus and vaccinated against MD compared with the chickens only vaccinated. Sixty-three one-day-old SPF chickens infected with adenovirus and vaccinated against MD were challenged with vv MD virus field strain. The protection index in this experiment was 55.6%-77.8%.
The strains of adenoviruses were isolated from 356 birds with clinical form of Marek’s disease and coinfection with adenoviruses. A hexon gene fragment coding loop L1 of adenovirus strains was sequenced and obtained data were analysed with BLAST, Geneious 5.3, and MEGA5 software by comparison with nucleotide sequences of reference strains of fowl adenoviruses (FAdV-1 - FadV-12), two turkey adenoviruses, and two goose adenovirus strains. On this basis, serotypes of adenovirus strains were determined. Sequences of all adenovirus strains isolated from birds infected with Marek’s disease virus were classified into six serotypes representing four species. Mostly FAdV-7, FAdV2/11, and FAdV-8a serotypes were found.
The study describes successful isolation of 96 fowl adenovirus (FAdV) strains from 789 chickens from 95 flocks. PCR specific for hexon gene encoding L1 loop was conducted. Amplicons were subjected to sequence analysis. The sequences were analysed by the software: BLAST, Geneious 6.0, and MEGA 5, then aligned with different adenovirus strain reference sequences accessible in GenBank database. The examined strains belong to the particular groups and serotypes. The sequences of all adenoviruses were classified into five species (FAdV A-E) and eight serotypes (FAdV-1, FAdV-2, FAdV-4, FAdV-5, FAdV-7, FAdV-8a, FAdV- 8b, and FAdV-11).
The aim of the study was to determine the influence of simultaneous infection of chicken embryo fibroblasts (CEF) with different doses of adenovirus field strain serotype 7 (FAdV-7 JN-5/10j) and turkey herpesvirus strain FC126 (FC126 HVT) on replication of the herpesvirus in in vitro cultures. Three experiments were performed: simultaneous infection of CEF with adenovirus and HVT; inoculation of CEF culture with adenovirus, followed by infection with HVT after 24 h; and inoculation of CEF with HVT, followed by the infection with adenovirus 24 h later. In order to detect the presence of HVT and adenovirus strains in CEF culture, SORF 1 and hexon genes were determined, respectively. The infection with adenovirus lowered replication of FC126 HVT in chicken embryo fibroblast.
Viral infections are the greatest threat to waterfowl and cause significant economic losses. Diagnosis and differentiation of three goose viruses is difficult in the field and often requires laboratory confirmation. Therefore, the aim of the study was to develop a triplex PCR and optimise its parameters for simultaneous detection of DNA of goose parvovirus (GPV), goose polyomavirus (GHPV), and goose circovirus (GoCV).
Material and Methods
The DNA of viruses isolated from field cases from the National Veterinary Research Institute’s own collection was used for the study. The primer attachment temperature, the number of reaction cycles, and the Taq DNA polymerase and Mg2+ concentrations were optimised. The sensitivity and specificity of this triplex PCR was also determined.
Based on the obtained results, triplex PCR parameters were optimised for simultaneous detection of DNA of GPV, GHPV, and GoCV in one sample. The following PCR products of the expected size were obtained: GPV DNA of 806 bp, GoCV DNA of 571 bp, and GHPV DNA of 180 bp.
The developed triplex PCR method proved to be useful for simultaneous detection of infections with three waterfowl viruses and will be used in relevant laboratory diagnostics.
Introduction: Avian reovirus (ARV) infections in poultry populations are reported worldwide. The reovirus belongs to the genus Orthoreovirus, family Reoviridae. The aim of the study was to evaluate the incidence of ARV infections in the poultry population based on diagnostic tests performed in 2010–2017.
Material and Methods: Samples of the liver and spleen were collected from sick birds suspected of ARV infection and sent for diagnostics. Isolation was performed in 5–7-day-old SPF chicken embryos infected into the yolk sac with homogenates of internal organs of sick birds. Four primer pairs were used to detect the σNS, σC, σA, and µA ARV RNA gene fragments. A nested PCR was used for the detection of the σNS and σC genes.
Results: In 2010–2017, ARV infection was found in birds from 81 flocks of broiler chickens and/or layers, 8 flocks of slaughter turkeys, and in 4 hatchery embryos at 17–20 days of incubation. The primers used in RT-PCR and nested PCR did not allow effective detection of ARV RNA in all virus-positive samples.
Conclusion: The problem of ARV infections in the poultry population in Poland still persist. The primers used for various ARV segments in RT-PCR and nested PCR did not allow effective detection of RNA in the visceral organs of sick birds. The presented results confirm the necessity of using classical diagnostic methods (isolation in chicken embryos, AGID).
Introduction: Avian poxvirus infections are widespread in the domestic poultry population but are also reported in wild birds. In poultry, these infections cause significant economic losses, while wild birds may be a reservoir for poxvirus which affects breeding poultry. However, wild birds may also exhibit characteristic anatomopathological changes. This study concerns the infection of wild-living great tits (Parus major) with the avian poxvirus in Poland.
Material and Methods: Samples of internal organs and skin collected from great tits were homogenised and total cellular DNA was isolated. In PCR, the primers complementary to gene encoding the core protein 4b of the HP44 strain of fowl poxvirus (FPV) were used.
Results: After electrophoresis in 2% agarose gel, the PCR product of 578 bp characteristic for FPV was obtained in DNA samples isolated from skin lesions and the heart. The analysis of the nucleotide sequence of the virus strain showed 99% similarity to many poxviruses previously isolated from great tits and other free birds at various sites in the world.
Conclusions: This paper is the first clinically documented evidence obtained in laboratory conditions of avian poxvirus cases in great tits in Poland.