Explore the vast range of titles available.

Background The incidence of reported cases of equine herpesvirus myeloencephalopathy (EHM) caused by infection with neuropathogenic strains of equine herpesvirus 1 (EHV-1) has markedly increased over the last decade in many Western countries. The purpose of this study was to estimate the prevalence of the neuropathogenic ( G2254 ) and non-neuropathogenic ( A2254 ) variants of EHV-1 among isolates associated with abortions in Polish stud farms. Results The results of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing were consistent, and showed that two out of 64 abortions (3.1%) were induced by the neuropathogenic genotype G2254 . All remaining 18 EHV-1 positive abortion cases (28.1%) were caused by the non-neuropathogenic genotype A2254 . Conclusions Most of the abortions in mares in Poland from 1999 to 2012 were associated with non-neuropathogenic strains of EHV-1. However, the presented data indicate that the neuropathogenic genotype of the virus is also present in Polish stud farms. Such a presence suggests that the future emergence of EHM in Poland is probable.

We used mass spectrometry-based protein identification to determine the presence of granins and other proteins in the mouse neuroblastoma secretome. We detected polypeptides derived from four members of the granin family: chromogranin A, chromogranin B, secretogranin III, and VGF. Many of them are derived from previously described biologically active regions; however, for VGF and CgB, we detected peptides not related to known bioactivities. Along with granins, we identified 115 other proteins secreted by mouse neuroblastoma cells, belonging to different functional categories. Fifty-six out of 119 detected proteins possess the signal fragments required for translocation into endoplasmic reticulum. Sequences of remaining 63 proteins were analyzed using SecretomeP algorithm to determine probability of nonclassical secretion. Identified proteins are involved in the regulation of cell cycle, proliferation, apoptosis, angiogenesis, proteolysis, and cell adhesion.

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.

Background Influenza virus isolation in embryonated chicken eggs (ECEs) is not applicable for rapid diagnosis, however it allows the recovery and propagation of the viable virus. A low number of infectious virus particles in the swabs, poor quality of samples or individual strain properties can lead to difficulties during the virus isolation process. We propose to utilize chorioallantoic membranes (CAM) of ECEs with the assistance of real-time RT PCR to facilitate equine influenza virus isolation. Methods Real-time RT PCR was used to detect influenza virus genetic material in amniotic/allantoic fluids (AF) and CAM of ECEs. Haemagglutination assay was used for AF. We used highly diluted virus as a substitute of clinical specimen for ECEs inoculation. Results Our study demonstrated that real-time RT PCR testing of CAM homogenates was more useful than testing of AF for EIV detection in ECEs. Positive results from CAM allowed to select the embryos from those with haemagglutination assay (HA) - and real-time RT PCR-negative AF for further passages. Using homogenates of CAM for subsequent passages, we finally obtained HA-positive AF, which confirmed virus replication. Conclusion We postulate that real-time RT PCR testing of CAM homogenates and their subsequent passages may facilitate the isolation of equine influenza viruses.

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.

•Beneficial results of using drug combination therapy.•Influence of using of INFα/β on rabies infection.•Extended of mice survival by inhibiting of RV replication. Rabies is a fatal disease of all mammals causing almost 60,000 human deaths every year. To date, there is no effective treatment of clinical rabies once the symptoms appear. Here, we describe the promising effect of combination therapy composed of molecules that target replication of the rabies virus (RV) at different stages of life cycle and molecules that inhibit some pathways of the innate host immune response accompanied by a blood-brain barrier opener on the outcome of RV infection. The study reports statistically significant extension of survival of mice treated with the drug cocktail containing T-705, ribavirin, interferon α/β, caspase-1 inhibitor, TNF-α inhibitor, MAPKs inhibitor and HRIG compared to the survival of mice in the virus control group (p = 0.0312). Furthermore, the study points to the significant impact of interferon α/β on the survival of RV-infected mice. We have shown a significant down regulation of pro-inflammatory molecules (caspase-1 and TNF-a) in the CNS in RV-infected mice treated with a combination of drugs including interferon α/β.

Here, we present the first detected cases of bluetongue virus (BTV) in native cattle from Poland. The virus was found in animals located near the Polish-Belarusian and Polish-Lithuanian borders. The positive animals were detected through an official epidemiological surveillance program. A combination of type-specific real-time RT-PCR and phylogenetic tests revealed the presence of BTV serotype 14 (BTV-14). This serotype is highly homologous to the vaccine strain and BTV-14 present in Russia, Lithuania, and Spain (from an animal imported from Lithuania). The most probable route of virus introduction to Poland was transmission through midges. All of the cases were subclinical.

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. Six rtRT-PCR/RT-PCR protocols were compared for the laboratory diagnostic of fourteen BTV-14 isolates circulating in Poland in 2012–2014. All 14 isolates were positive in the protocols of Shaw (18), a commercial LSI NS3 kit, and Eschbaumer (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 (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. The paper points to the possible false negative results in the diagnosis of BTV infections depending on the protocol used.

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. 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. 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. 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.

The aim of the study was to estimate and compare the distribution of Culicoides biting midges species at farms with different main hosts — cattle and horse. Culicoides spp. are known vectors of arboviruses including African horse sickness virus (AHSV), bluetongue virus (BTV) and Schmallenberg virus (SBV). The latter two have been already reported in Polish ruminants recently, while AHSV remains absent, however the risk of its emergence has been increasing in the recent years. In order to establish the activity of potential AHSV vector at vicinity of horses, an OVI midge trap has been placed at the horse stables in the southeastern Poland. Another trap has been placed 7 km away at the cattle farm. The collections were carried over the midge activity season from April until November 2016. The midge abundances at both sites were comparable with the total numbers of insects trapped of 43,153 and 34,829 at the cattle and horse farm, respectively. Midges belonging to C. obsoletus/scoticus complex were the dominant ones at both locations. The other most abundant species were C. punctatus and C. pulicaris , while the other ten species identified ( C. chiopterus , C. deltus , C. dewulfi , C. fagineus , C. impunctatus , C. newsteadi , C. nubeculosus , C. parroti , C. riethi , C. stigma ) accounted for less than 0.5%. The study has shown that the Orbivirus vectors are present at a high abundance at the Polish horse farm, what may be a helpful tool in the AHS risk assessment in the nonendemic part of Europe.