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  • Author: Jan F. Żmudziński x
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Open access

Marcin Smreczak, Anna Orłowska, Paweł Trębas and Jan F. Żmudziński

Abstract

The paper describes the data concerning rabies in domestic animals and in wildlife as well as in bats in Poland in 2009 and 2010. Analysis of rabies situation was based on species involved and geographical distribution of rabies outbreaks. Favourable decreasing trend in rabies epidemic in 2009 was stopped by the outbreak of rabies in the Malopolska province in 2010. This resulted in dramatic increase in the number of rabies cases. Emergency vaccination in the zone of rabies outbreak with increased number of vaccines per km2 in bordering areas of the province has improved epizootic situation, which returned to the state before the outbreak. To monitor rabies situation a strict supervision of all elements of the ORV and surveillance of rabies is necessary.

Open access

Aleksandra Kuta, Mirosław P. Polak, Magdalena Larska and Jan F. Żmudziński

Abstract

The aim of the study was to evaluate the status of bovine viral diarrhoea virus (BVDV) infection in selected dairy herds in Poland with the use of commercial enzyme linked immonosorbent assay for the detection of specific antibodies (BVDV-Ab ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) for the detection of viral RNA, using bulk tank milk (BTM) samples. Two hundred and thirty-one samples of BTM were collected from 99 dairy herds in Poland. The herds were divided into four different classes according to the Swedish system of classification. The results showed that 70.7% of herds were BVDV antibodypositive. High levels of antibodies in 52.85 % (37 herds in class 3) of all antibody positive herds indicated acute BVDV infection. Thirty five samples with the highest antibody levels were tested by RT-PCR and five of them were positive for viral RNA. Dairy herds in Poland have high levels of antibodies against BVDV in BTM. Since no vaccination was implemented in the herds tested, high seroprevalence of BVDV antibodies in cattle indicates the widespread of BVDV infection in Polish cattle.

Open access

Mirosław P. Polak, Aleksandra Antos, Jerzy Rola and Jan F. Żmudziński

Abstract

Introduction: Bovine viral diarrhoea (BVD), caused by the bovine viral diarrhoea virus (BVDV), is one of the most important diseases of cattle worldwide. The purpose of the study was to determine the BVDV infection status in a dairy herd vaccinated against BVD. Before vaccination started in 2008, there had been no prior identification or the removal of the possible source of infection (persistently infected animals). It was expected that vaccination itself would enable the elimination of viral shedders on a long term basis. Material and Methods: Serological screening for antibodies against BVDV with determination for antibodies titres, BVDV antigen, and the presence of the viral genome with phylogenetic analysis of positive samples in the herd were performed, despite the lack of any clinical problems indicating possible presence of BVDV infection. Results: 19 individuals persistently infected with BVDV were identified among calves and heifers but not in adult cattle. All virus shedders were antibody negative and the genotype of isolated virus was BVDV-1b, indicating a single source of infection. The vaccine used in the herd was composed of BVDV-1a strain. In each of the tested cowsheds, antibody titres against BVDV-1b were higher than against BVDV-1a (median values). Conclusion: Despite a long-lasting vaccination programme and relatively high sequence homology of vaccinal and field strains of BVDV (83.6%), it was not possible to avoid transplacental infections of foetuses and the birth of persistently infected calves from vaccinated heifers although the protection against clinical disease was accomplished.

Open access

Anna Orłowska, Jan F. Żmudziński, Marcin Smreczak, Paweł Trębas and Anna Marzec

Abstract

Introduction: The reverse transcription polymerase chain reaction (RT-PCR) is one of the most extensively used methods for identification of animals infected with bluetongue virus (BTV). There are several RT-PCR protocols published and several real-time RT-PCR (rtRT-PCR) commercial kits available on the market. Because Poland faced BTV-14 infection in 2012, different protocols were implemented in the country to confirm the RT-PCR results positive for this virus. The article presents a comparative study of several RT-PCR protocols and discusses their diagnostic reliability and applicability.

Material and Methods: Six rtRT-PCR/RT-PCR protocols were compared for the laboratory diagnostic of fourteen BTV-14 isolates circulating in Poland in 2012–2014.

Results: All 14 isolates were positive in the protocols of Shaw et al. (18), a commercial LSI NS3 kit, and Eschbaumer et al. (5). Four out of fourteen BTV-14 isolates gave positive results in Hoffmann’s 2 and 6 protocols and none of the 14 isolates yielded positive results in Maan et al. (8) method. Phylogenetic study of a short fragment of 450 nt of BTV segment 2 (258–696 positions) revealed 100% identity within Polish variants and with Russian and Spanish isolates.

Conclusion: The paper points to the possible false negative results in the diagnosis of BTV infections depending on the protocol used.

Open access

Aleksandra Kuta, Mirosław P. Polak, Magdalena Larska and Jan F. Żmudziński

Abstract

Restriction fragment length polymorphism (RFLP) analysis was developed for genetic typing of Polish strains of bovine viral diarrhoea virus (BVDV). The method was applied using 60 BVDV isolates, which included BVDV genotype 1, subtypes a, b, d, e, f, and g, and genotype 2a. RT-PCR products of the 5’untranslated region (5’UTR) were digested using three enzymes. Restriction patterns classified the strains into seven groups, each with a specific and different pattern from other subtypes. These findings were confirmed by nucleotide sequencing and phylogenetic analysis. The results suggest that RFLP analysis is a simple, reliable, and fast genotyping method for BVDV strains in comparison with sequencing. This method can distinguish six subtypes of BVDV-1 including a new subtype 1e, identified exclusively by this method, and it allows differentiation of BVDV-1 from BVDV-2 genotype.

Open access

Małgorzata Kwaśnik, Wojciech Rożek and Jan F. Żmudziński

Abstract

The purpose of the experiment was to compare apoptosis induced by equine influenza virus (EIV A1 and EIV A2) infection in MDCK, RK13, and NEURO-2A cell lines. Flow cytometry was used to observe two symptoms of apoptosis: phosphatidylserine translocation in plasmalemma (annexin V assay) and the fragmentation of DNA generated by endonuclease activity (TUNEL assayterminal deoxynucleotidyl transferase biotin-dUTP nick end labelling). The differences in the onset of apoptosis in the studied cells was observed. In MDCK cells infected with EIV A1 and A2, a weak signal of the phosphatidylserine translocation was observed but more cells showed the DNA fragmentation. An opposite effect was observed in case of RK 13 cells. NEURO-2A cells displayed a similar number of annexin V and TUNEL positive cells after the infection with EIV A2, while in case of EIV A1 infection, only the early symptoms of apoptosis were noted. Differences between both viral serotypes could originate from functioning of viral proteins responsible for induction or inhibition of apoptosis. The differences between cell types may result from the activation of cellular pro or anti-apoptotic mechanisms.

Open access

Jerzy Rola, Wojciech Socha and Jan F. Żmudziński

Abstract

The variability of the ORF2a, ORF2b, ORF3, and ORF4 genes of the equine arteritis virus (EAV) was analysed during a seven year observation of persistent infection in a stallion of the Malopolska breed. A total of 11 semen samples were collected between 2004 and 2011. RNA of EAV isolates obtained from the semen of the stallion was amplified, sequenced, and compared with the sequences of other strains available in GenBank. Multiple nucleotide substitutions were found in sequences of the analysed regions, however, neither deletion nor insertions were detected. The highest number of point mutations (11-6 synonymous and 5 non-synonymous) were found in the ORF2b gene, and the lowest number of substitutions (6-5 synonymous and one non-synonymous) were found in the ORF2a gene. None of the identified mutations affected any of the glycosylation or phosphorylation sites of the minor EAV protein. Phylogenetic analysis of the ORF3 gene of EAV isolates showed that they grouped together within the cluster of European strains of EAV. Additionally, the ORF3 gene sequences of the isolates showed high (86.4% - 98.3%) similarity to the previously isolated Polish EAV strains.

Open access

Wojciech Socha, Jerzy Rola, Dariusz Bednarek, Renata Urban-Chmiel and Jan F. Żmudziński

Abstract

Shedding time of bovine respiratory syncytial virus (BRSV) and bovine parainfluenza virus 3 (BPIV3) in calves vaccinated intranasally with modified live Rispoval RS-PI3 vaccine was determined. Blood and nasal swabs were collected on selected days before and after vaccination. Antibodies against BRSV and BPIV3 were tested by Respiratory ELISA Pentakit and the viral RNA was detected by RT-PCR. Twenty eight days after administration of the vaccine, a marked increase of specific antibody titres to BRSV and BPIV3 was detected in vaccinated calves. All animals were RT-PCR positive both for BRSV and BPIV3. Both viruses were excreted with nasal discharges within 8 d after vaccination but the course of shedding in individual calves was variable.

Open access

Małgorzata Kwaśnik, Ilona M. Góra, Jan F. Żmudziński, Jerzy Rola, Mirosław P. Polak and Wojciech Rożek

Abstract

Introduction

Traditionally, evolutionary analysis of equine influenza virus (EIV) is based on the HA gene. However, the specificity of the influenza virus enables the classification of viral strains into different phylogenetic groups, depending on the gene being analysed. The aim of the study was to analyse phylogenetic paths of EIV based on M gene with reference to the HA gene.

Material and Methods

M gene of Polish isolates has been sequenced and analysed along with all M sequences of EIV available in GenBank database. Phylogenetic analysis was performed using BioEdit, ClustalW, and MEGA7 softwares.

Results

The clustering of the strains isolated not only from Asia but also from Europe into one common Asian-like group of EIV was observed. Twelve nucleotide substitutions in the M gene of strains from the Asian-like group were crucial for the evolutionary analysis. We also observed homology in the M gene of the Asian-like and H7N7 strains.

Conclusions

M gene specific for the Asian-like group is present in strains recently isolated in Europe and Asia, which were classified previously in the Florida 2 clade based on HA. Therefore, Asian-like group does not seem to be assigned to a specific geographical region. Traces of H7N7 strains in more conservative genes like M of some contemporary EIV strains may indicate the link between the old phylogenetic group and recent H3N8 strains. Analysis of conservative genes may be more useful in tracking the direction of virus evolution than in the genes where the high variability rate may blur the original relationships.