Aims: Riemerella anatipestifer is a significant pathogen of waterfowl and turkeys. Due to their similar ecology and morphological and cultural characteristics it is important to differentiate R. anatipestifer infections from those caused by Pasteurella multocida. Present study describes a novel PCR assay that is capable of rapid and species‐specific identification of R. anatipestifer from bacterial cultures.
Methods and Results: An ERIC (enterobacterial repetitive intergenic consensus)‐PCR fragment common to all tested isolates was used as a target for primer design. After optimization, the assay was tested on 72 R. anatipestifer strains isolated from clinical samples and identified using biochemical tests. All of these gave positive results, while heterologous pathogens, including different serotypes of P. multocida, proved to be negative. The assay was also capable of demonstrating R. anatipestifer directly from five clinical samples.
Conclusions: The presented PCR is suitable for proper identification of R. anatipestifer from culture. Preliminary investigation showed that the test could be suitable for detection of the pathogen from clinical samples as well.
Significance and Impact of the Study: The described PCR assay will improve the fast and proper identification of R. anatipestifer.
In this work, we examined the diversity of fowl adenovirus (FAdV) types occurring in Hungary. From diseased chicken flocks in Eastern Hungary, 29 FAdV strains were isolated between 2011 and 2015. We performed molecular typing of the isolates based on their partial hexon sequences. The results showed that representatives from every FAdV species from A to E are present in Hungary, but compared to the findings from our previous survey, a lower number of different FAdV types were detected. Inclusion body hepatitis was always associated with FAdV-2 or -8b, gizzard erosion was caused in almost every case by FAdV-1. Numerous strains belonging to species FAdV-B were found. The complete genome sequence of a candidate new genotype strain, showing the highest divergence from the reference FAdV-5, was determined using next generation sequencing. In order to provide results compatible with the serology-based type classification, multiple genomic regions, including the major antigenic determinants, of the new isolate (strain 40440-M/2015) were compared to their counterparts in the prototype FAdV-5 (strain 340) from species FAdV-B, at both nucleotide and amino acid sequence levels. In different comparative analyses, the two strains were always found to have larger divergence between each other than any two of the most closely related FAdV serotypes. This new emerging FAdV genotype is already present in Hungary and Austria, though its exact pathological role requires further investigations. The introduction of a novel FAdV (geno)type for the classification of these strains is further supported.
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