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  • Author: Krzysztof Śmietanka x
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Open access

Edyta Świętoń and Krzysztof Śmietanka


Introduction: The genomes of nine H5 subtypes of low pathogenic avian influenza virus (LPAIV) strains identified in wild birds in Poland between 2010 and 2015 were sequenced, and their phylogenetic relationship was determined.

Material and Methods: AIV genome segments were amplified by RT-PCR and the PCR products were sequenced using Sanger method. Phylogenetic trees were generated in MEGA6 software and digital genotyping approach was used to visualise the relationship between analysed strains and other AIVs.

Results: High genetic diversity was found in the analysed strains as multiple subgroups were identified in phylogenetic trees. In the HA tree, Polish strains clustered in two distinct subclades. High diversity was found for PB2, PB1, PA and NP, since 5-8 sublineages could be distinguished. Each strain had a different gene constellation, although relationship of as much as six out of eight gene segments was observed between two isolates. A relationship with poultry isolates was found for at least one segment of each Polish strain.

Conclusion: The genome configuration of tested strains indicates extensive reassortment, although the preference for specific gene constellation could be noticed. A significant relationship with isolates of poultry origin underlines the need for constant monitoring of the AIV gene pool circulating in the natural reservoir.

Open access

Anna Pikuła, Katarzyna Domańska-Blicharz, Rytis Cepulis and Krzysztof Śmietanka


Introduction: Infectious bursal disease virus (IBDV) is a causative agent of immunosuppressive disorder resulting in significant losses to the world poultry industry. This study describes the molecular characterisation of an atypical IBDV from a field outbreak that occurred in vaccinated chicken flocks in Latvia in 2011.

Material and Methods: Ten bursae of Fabricius from each flock were collected for laboratory examination. Virus isolation was performed in embryonated eggs and CEF culture. The RT-PCR aimed at hypervariable domain of VP2 gene combined with sequencing was performed for detection and identification of IBDV.

Results: The molecular examinations confirmed the IBDV infection. The analysis of the amino acid sequence revealed that the strain possessed four amino acids at VP2 protein (222A, 256I, 294I, and 299S), indicating a genetic relatedness to a very virulent IBDV. However, some unique or rare amino acid substitutions (219L, 220F, 254D, 279N, and 280T) were also detected.

Conclusion: The obtained results demonstrate the occurrence of IBDV with a high mutation rate within the hypervariable domain of VP2 peptide, and highlight the necessity of implementation of IBDV surveillance in Eastern European poultry industry to determine whether this strain is an exception or a new wave of IBDV with new genetic features emerged in the field.

Open access

Monika Olszewska-Tomczyk, Izabella Dolka, Edyta Świętoń and Krzysztof Śmietanka



Genotype VI of avian avulavirus 1 (AAvV-1) has pigeons and doves as its reservoir and is often termed pigeon paramyxovirus type-1 (PPMV-1). The pathogenesis of PPMV-1 infections in poultry is largely obscure. It is known that PPMV-1 requires a series of passages in chickens before it becomes adapted to gallinaceous poultry.

Material and Methods

Changes in the genome of PPMV-1 were analysed after serial passages in specific pathogen free (SPF) chickens, using high-throughput sequencing. Additionally, histopathological lesions induced by PPMV-1 in experimentally inoculated pigeons, chickens, and turkeys were evaluated.


Following six passages of PPMV-1 in chickens, 10 nonsynonymous substitutions were found including one (in the NP protein) which dominated the genetic pool of viral quasispecies. Histopathological changes induced by the post-passage PPMV-1 strain were more prominent than changes wrought by the pre-passaged PPMV-1 strain and the lesions were most intense in pigeons followed by chickens and turkeys.


PPMV-1 is highly adapted to pigeons and passaging through chickens results in the acquisition of novel amino acids in the polymerase complex, which may alter the pathogenic potential of the virus.

Open access

Michał Jóźwiak, Krzysztof Wyrostek, Katarzyna Domańska-Blicharz, Monika Olszewska-Tomczyk, Krzysztof Śmietanka and Zenon Minta


Introduction: The aim of the study was to test the utility of Flinders Technology Associates filter paper (FTA® Cards) for molecular detection and storage of avian influenza virus (AIV). Material and Methods: There were two strains of AIV used in the study: low pathogenicity H7N1 and high pathogenicity H5N1 subtypes. Detection of viral material was conducted using molecular RT-PCR and rRT- PCR method. Results: The infectivity of LPAIV/H7N1 and HPAIV/H5N1 was completely inactivated within 1 h and 24 h after adsorption to FTA® Cards at room temperature, respectively. Viruses stored on FTA® Cards had detection limit approximately 1 log10 lower than live viruses. Viral RNA of both strains were detectable on the cards by rRT-PCR for a minimum of 150 d, irrespectively of storage temperatures (room temperature, -20ºC). RNA was also detected in all samples obtained from SPF chickens experimentally infected with HPAI/H5N1 on 3rd and 4th day post-infection (p.i.).

Conclusion: FTA® Cards enable safe and effective alternative transport of samples for molecular diagnosis of AIV.