A complete analysis of Relative Synonymous Codon Usage in HVRs1-4 region in adenovirus genome.

Recent outbreaks of adenovirus (FAdV) infections in poultry flocks have been determined in many countries in Europe, Asia and Australia connected with economic consequences, and loses in poultry production. To better understand the evolution and transmission of FAdV viruses, de- tailed codon usage analysis was performed for 137 recently obtained FAdV strains. A high effec- tive number of codons, and an indication the presence of low codon usage were determined. The presence of mutations, and their influence on codon usage was confirmed by a correlation be- tween nucleotide compositions at the 3rd codon positions, HVRs1-4, and ENCs. This presence indicate some influence of natural selection, and antigenic properties of examined FAdV strains.

The development and evaluation of cross-priming amplification for the detection of avian reovirus.

AIMS: The aim of this study was to develop and evaluate cross-priming amplification (CPA) for the detection of avian reovirus (ARV). METHODS AND RESULTS: Five specific primers were designed, on the basis of the σNS sequence of the S1133 ARV strain. Incubation temperature and primer concentrations were determined. The optimal incubation conditions in a water bath were 61.3°C for 45 min. No reverse transcription stage was required. The results were recorded under UV light illumination as a bright, greenish fluorescence in positive samples, and through the lack of this in negative controls and samples. Additionally, the gel electrophoresis performed during analysis showed the presence of ladder-like patterns, formed by hairpin-like CPA products. The developed CPA method was compared to reverse-transcription polymerase chain reaction (RT-PCR) and real-time RT-PCR. Sensitivity of CPA was estimated using seven dilutions of standard S1133 strain and reached 0.05 log10 TCID50 ml(-1). RT-PCR sensitivity reached 2.5 log10 TCID50 ml(-1) and was 1000 times lower than for CPA, whereas real-time RT-PCR sensitivity reached 1.5 log10 TCID50 ml(-1). Analysis of 32 RNAs extracted from field specimens showed the presence of an ARVσNS fragment in 4 (12.5%) samples. Interestingly, the positive samples originated from flocks affected by Marek's disease (MD) or fowl adenovirus (FadV). RT-PCR was unable to detect ARV, due to its lower sensitivity. However, the real-time RT-PCR that was conducted confirmed the CPA study. CONCLUSIONS: CPA is a very sensitive and rapid method, which allows ARV detection using simple laboratory equipment. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the application of the CPA method for detection of ARV, using simple laboratory equipment.

Interactions between Marek's disease virus Rispens/CVI988 vaccine strain and adenovirus field strain in chicken embryo fibroblast (CEF) cultures.

The aim of the study was to determine the influence of adenovirus infection on the replication of Marek's disease virus vaccine strain Rispens/CVI988 during in vitro co-infection studies. Adenovirus field strain JN-5/10j was isolated from sick chickens. The study was conducted in chicken embryo fibroblast cultures (CEF). Monolayers of CEFs were infected with Rispens strain and field adenovirus strain JN-5/10j with different doses (10(1.0)-10(3.0) TCID50) in the following manner: a) simultaneously, b) first, infection with Rispens strain and after 24 h infection with adenovirus strain JN-5/10j and c) infection with adenovirus strain JN-5/10j 24 h before infection with Rispens strain. After 18, 24, 48, 72, and 96 h of incubation, the copy number of the pp38 gene of Rispens strain was determined using Real-time PCR. The results indicated that the Adenovirus infection before the infection with Rispens strain reduced the replication of the pp38 gene after 48 h by 2 log10.

The survey of wild birds for West Nile virus in Poland.

Two thousand one hundred and forty birds belonging to 39 different species from different locations in Poland were examined. The study has taken place from the early spring till late autumn 2007-2010 when the activity of the mosquitoes was the highest. The brain samples were taken from the birds and whole cellular RNA was isolated, then the RT-PCR and NRT-PCR were performed to detect the presence of West Nile virus (WNV). The obtained results were confirmed by the commercial WNV Kit. No genetic material of WNV was found in the examined samples.