Incidence of Chlamydia spp., FIV, FeLV in Free-Roaming Cats in Slovakia.

Purpose: Free-roaming cats represent a potential reservoir of infectious diseases. The most common co-infections of free-roaming cats include mixed viral, bacterial, fungal, yeast and parasitic infections. This study focuses on the occurrence of Chlamydia spp. feline immunodeficiency virus (FIV), feline leukaemia virus (FeLV) and their co-infections. The diseases accompanied by immune suppression, such as FIV, create favourable conditions for the onset of other diseases and co-infections. The result of co-infection may be a higher susceptibility for other pathogens, as well as the occurrence of more severe clinical symptoms. Patients and Methods: The study involved 168 (113♀ and 55♂) free-roaming adult cats during the years 2021-2022. All cats belonged to Slovak citizens with permanent residence in the Slovak Republic. Blood samples and swabs (Invasive EUROTUBO® Collection sterile swab, Deltalab O8191 Rubí, Spain) from the conjunctival sac were taken from 168 cats to be later tested by PCR and ELISA methods. Statistical analysis was also performed. Results: The overall prevalence of Chlamydia spp. was 17.26%, of FIV 15.48%, and 5.95% of FeLV. The most significant finding in our study was 3.57% co-infection of FIV and Chlamydia spp. in tested cats. Conclusion: The observed prevalence of Chlamydia spp. FIV and FeLV indicates that the presence of these pathogens in populations of free-roaming cats is endemic.

The first evidence of Asian-like CPV-2b in Slovakia in a vaccinated dog with an acute fatal course of parvovirus infection: a case report.

This study provides a comprehensive description of the clinical course of a fatal parvovirus infection in a vaccinated dachshund puppy, along with the first identification of a new CPV-2 variant in Slovakia, elucidated through molecular amino acid analysis of the VP2 gene. The dog exhibited clinical signs such as apathy, vomiting, and bloody diarrhea. After confirming CPV-2 infection with a commercial snap test, intensive therapy was initiated. The dog succumbed within 48 h of admission. A rectal swab sample was collected, CPV-2 was examined using the PCR method, and sequenced. The virus detected in the patient was related to strains of CPV-2c of Asian origin and unrelated to European CPV-2b strains. The sequence had genetic signatures typical of Asian strains (VP2: 5Gly, 267Tyr, 324Ile, 370Arg, and 440Thr). Phylogenetic analysis classified this strain as similar to Asian strains of CPV-2c. It is believed to be derived from an Asian strain similar to CPV-2c that acquired the 426Asp mutation. With this finding, we present the first evidence of an Asian-like CPV-2b strain in the territory of Slovakia.

Investigation of canine parvovirus occurrence in cats with clinical signs of feline panleukopenia in Slovakia - pilot study.

Introduction: Feline panleukopenia is a contagious viral disease caused by the feline panleukopenia virus (FPV). A closely related pathogen is canine parvovirus (CPV), and amino acid substitutions in this virus allow it to acquire a feline host range. In feline hosts, the disease induced by CPV manifests with similar symptoms to those caused by FPV or milder ones, leading to its underdiagnosis. The aim of this study was to determine the presence of CPV type 2 (CPV-2) in cats with clinical symptoms of panleukopenia and to assess the use of commercial CPV antigen tests for the clinical diagnosis of FPV. Material and Methods: Samples from 59 cats from central Slovakia were included in the study. Rectal swabs were collected and clinically tested for parvovirus infection using a commercial antigen test. Antigen-positive samples were confirmed by PCR targeting the viral VP2 gene. The sequences of the PCR products were established with the Sanger method. Results: Of 59 samples, 23 were revealed to be positive for parvovirus infection by both antigen and PCR test (38.9%). Analysis with the National Center for Biotechnology Information BLASTn application showed 99.78-100% pairwise identity with FPV. The mortality rate of parvovirus-infected cats included in this study was 8.69% (2/23). Conclusion: Although feline disease with CPV-2 was not confirmed, the CPV antigen test was able to detect FPV infection.

Opportunities and Limitations of Molecular Methods for Studying Bat-Associated Pathogens.

Bats have been identified as reservoirs of zoonotic and potentially zoonotic pathogens. Significant progress was made in the field of molecular biology with regard to infectious diseases, especially those that infect more than one species. Molecular methods, sequencing and bioinformatics have recently become irreplaceable tools in emerging infectious diseases research and even outbreak prediction. Modern methods in the molecular biology field have shed more light on the unique relationship between bats and viruses. Here we provide readers with a concise summary of the potential and limitations of molecular methods for studying the ecology of bats and bat-related pathogens and microorganisms.

Serological survey of lyssaviruses in synanthropic bats and human exposure to bats in Slovakia.

INTRODUCTION: Bats are considered natural reservoirs for lyssaviruses. A total of 17 out of 19 known lyssaviruses circulate in bat populations. Lyssaviruses cause rabies in animals and humans. The transmission of lyssaviruses from European bats to terrestrial animals and humans is rare, but the risk of infection still exists even in developed countries. Slovakia is currently a rabies-free country. OBJECTIVE: The aim of the study was to assess the potential circulation of EBLV-1 in synanthropic bats present in human inhabited buildings, and to give an overview of human exposure to bats. MATERIAL AND METHODS: A passive serological survey targeted the prevalence of antibodies to bat lyssaviruses in synanthropic bats between 2009 - 2019. A total of 598 bats of the species Pipistrellus pipistrellus, Pipistrellus pygmaeus, Eptesicus serotinus, Nyctalus noctula and Vespertilio murinus were captured in buildings mainly in Eastern Slovakia, and examined by the rapid fluorescent focus inhibition test (RFFIT). RESULTS: Lyssavirus-specific antibodies were detected in 2 (0.3%) of the 598 examined bats. Additionally, brain tissues of bats found dead were examined using the standard fluorescent antibody test (FAT) with negative results. An overview of available data on human exposure to bats recorded in Slovakia from 2007 - 2019 is also included. CONCLUSIONS: The study confirmed the presence of lyssavirus antibodies in synanthropic bats in Slovakia, suggesting the active circulation of bat lyssaviruses in bat populations exploiting human buildings. Although the seroprevalence was found to be extremely low, the results show that any case of human exposure to bats must be treated with caution in order to protect public health.

The first serological evidence of Anaplasma phagocytophilum in horses in Slovakia.

Anaplasma phagocytophilum is the causative agent of granulocytic anaplasmosis. It affects humans and several wild and domesticated mammals, including horses. The aim of our study was a preliminary survey of the occurrence of these re-emerging pathogens in horses in Slovakia. The sera from 200 animals of different ages and both sexes were tested for the presence of A. phagocytophilum antibodies by indirect immunofluorescence assay. Subsequently, detection of the 16S rRNA gene fragment of A. phagocytophilum was attempted by polymerase chain reaction (PCR) in each blood sample. Our results confirmed the presence of specific antibodies in 85 out of 200 individuals (42.5%), but no significant changes were found between the animals of different ages and sexes. However, the PCR analysis did not detect any positive animals. Our data represent one of the highest values of seropositivity to A. phagocytophilum in horses in Central Europe. These results may contribute to a better understanding of the circulation of A. phagocytophilum in this region, thus indicating a potential risk to other susceptible species.

Transcriptome analysis of human brain microvascular endothelial cells response to Neisseria meningitidis and its antigen MafA using RNA-seq.

Interaction of Neisseria meningitidis (NM) with human brain microvascular endothelial cells (hBMECs) initiates of multiple cellular processes, which allow bacterial translocation across the blood-brain barrier (BBB). NM is equipped with several antigens, which interacts with the host cell receptors. Recently we have shown that adhesin MafA (UniProtKB-X5EG71), relatively less studied protein, is one of those surface exposed antigens that adhere to hBMECs. The present study was designed to comprehensively map the undergoing biological processes in hBMECs challenged with NM or MafA using RNA sequencing. 708 and 726 differentially expressed genes (DEGs) were identified in hBMECs exposed to NM and MafA, respectively. Gene ontology analysis of the DEGs revealed that several biological processes, which may alter the permeability of BBB, were activated. Comparative analysis of DEGs revealed that MafA, alike NM, might provoke TLR-dependent pathway and augment cytokine response. Moreover, both MafA and NM were able to induce genes involved in cell surface modifications, endocytosis, extracellular matrix remodulation and anoikis/apoptosis. In conclusion, this study for the first time describes effect of NM on the global gene expression in hBMECs using high-throughput RNA-seq. It also presents ability of MafA to induce gene expression, which might aid NM in breaching the BBB.

Comparative genomic hybridization in detection of DNA changes in canine lymphomas.

In this study, chromosomal imbalances in tumor tissues (lymphomas) and nucleotide changes in tumor suppressor TP53 were studied in a Bernese Mountain dog bitch and a cross breed bitch. Using comparative genomic hybridization, numerous chromosomal rearrangements were detected, which indicated the heterogeneity in tumor growth: in the cross breed bitch, a deletion on the chromosome 9, and duplications on chromosomes 5, 8 and 17 have been found. In the Bernese Mountain Dog bitch, losses on chromosomes 1, 5, 8, 12, 18, 22, 27, 29 and gains on chromosomes 1, 2, 9, 11, 15, 16, 18, 20, 23, 24, 25, 28, 29, 30, 34, 36, 37 and 38 were identified. With the sequencing of the TP53 gene, one silent mutation, transition A/G at position 138 in exon 5 was detected, without changing the amino acid.

Evaluation of potential genotoxic/cytotoxic effects induced by epoxiconazole and fenpropimorph-based fungicide in bovine lymphocytes in vitro.

Potential genotoxic/cytotoxic effects of the epoxiconazole/fenpropimorph-based fungicide were investigated using single cell gel electrophoresis and cytogenetic assays: chromosomal aberrations, sister chromatid exchanges, micronuclei and fluorescence in situ hybridization in cultured bovine lymphocytes. No statistically significant elevations of DNA damage and increases in cytogenetic endpoints were seen. However, evident cytotoxic effect presented as a decrease in mitotic and proliferation indices were recorded after exposure of bovine lymphocytes to the fungicide for 24 and 48 h at concentrations ranging from 3 to 15 µg mL(-1) (P < 0.05, P < 0.01, P < 0.001). Similarly, for 24 h an inhibition in the cytokinesis block proliferation index (CBPI) was obtained after exposure to the fungicide at concentrations ranging from 1.5 to 15 µg mL(-1) (P < 0.01, P < 0.001) in each donor.

The effect of thiacloprid formulation on DNA/chromosome damage and changes in GST activity in bovine peripheral lymphocytes.

The potential genotoxic effect of thiacloprid formulation on bovine peripheral lymphocytes was evaluated using the comet assay and the cytogenetic endpoints: chromosome aberrations (CAs), sister chromatid exchanges (SCEs) and micronuclei (MNi). Whole blood cultures were treated with the insecticide at concentrations of 30, 60, 120, 240 and 480 µg mL(-1) for 24, 48 h and/or 2 h of incubation. A statistically significant increase in the frequency of DNA damage, as well as in unstable chromosome aberrations (% breaks) were found after exposure to the insecticide at concentrations ranging from 120 to 480 µg mL(-1) (P < 0.05, P < 0.01, P < 0.001). For the detection of stable structural chromosome aberrations (e.g., translocations) and numerical aberrations by the FISH method, three whole chromosome painting probes for bovine chromosomes 1, 5 and 7 (BTA1, BTA5 and BTA7) were used in our experiments. We observed numerical aberrations, but without any statistical significance. Regarding the sister chromatid exchanges, no significant elevation in the SCE frequencies was found after 24-h exposure to the insecticide. A dose-related response in the SCE induction was obtained in bovine cultures after the prolonged time of exposure (48 h) to thiacloprid formulation at concentrations ranging from 120 to 480 µg mL(-1) in each donor (P < 0.05, P < 0.01), which was associated with a reduction of the PI (P < 0.05, P < 0.01). The insecticide failed to produce MNi; however, a significant reduction of CBPI was observed. Using real-time PCR, a decrease in the expression of bovine glutathione S-transferase M3 (GSTM3) was detected at the lowest dose. The higher concentrations of thiacloprid formulation caused an increase in the mRNA expression.