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.

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.

Ultra-high performance hydrophilic interaction liquid chromatography - Triple quadrupole tandem mass spectrometry method for determination of cysteine, homocysteine, cysteinyl-glycine and glutathione in rat plasma.

An ultra-high performance hydrophilic interaction liquid chromatography - triple quadrupole tandem mass spectrometry method was developed for the determination of biologically important thiols, namely cysteine, homocysteine, cysteinyl-glycine, glutathione, in rat plasma. The sample preparation procedure as well as the analytical method were comprehensively optimized and subsequently validated. An optimum sample preparation protocol was based on the simple and fast derivatization of the thiols with new derivatization reagent, N-phenylmaleimide, enabling highly selective and sensitive quantification in plasma matrices. The method, characterized by favourable performance parameters and meeting the FDA criteria for biomedical analysis, was successfully applied for monitoring the concentration levels of the selected thiols in the samples from transgenic rat model for tauopathy. The study revealed significant changes in homocysteine and glutathione levels related to tauopathy while other thiols did not indicate such relationship. Indeed, these findings could play an important role in further understanding of tauopathy process in the brain. Moreover, the proposed highly effective, reliable and robust analytical protocol can be easily adapted for other thiol compounds, spreading its application range in this biomedical field.

A frequent roundworm Baylisascaris transfuga in overpopulated brown bears (Ursus arctos) in Slovakia: a problem worthy of attention.

The genus Baylisascaris (order Ascaridida) includes numerous relatively host-specific nematodes, which are common in intestines of wild mammals. Some of them may have impact on veterinary and public health, as their larvae have the potential to cause visceral, ocular, and/or neural larva migrans in a wide range of mammals, birds, and humans. Baylisascaris transfuga is a parasite occurring in a range of bear species throughout the world. We present the current data on B. transfuga occurrence in brown bears from a relatively restricted territory of the Polana Protected Landscape Area in Central Slovakia, obtained by traditional methods (faecal examination, morphology). Species affiliation was confirmed by employing molecular markers generating nuclear 28S and mitochondrial cox1 sequences in adult worms. Based on 17 examined samples (15 excrements and two intestines of young bear females), the occurrence of B. transfuga in the surveyed area was assessed as 52.9%. Both bear females were infected with adult and juvenile worms. Due to the high density of bears in the locality, the high infection rate with ascarids, and the huge number of eggs produced by the parasites, it is apparent that the respective environment, including the inhabited areas, might be markedly contaminated by Baylisascaris eggs. The ability of B. transfuga to serve as a zoonotic agent has not been unambiguously proved; however, this attribute should be considered and subjected to further research.

Dog's genotype of Giardia duodenalis in human: first evidence in Europe.

The unicellular parasite Giardia duodenalis has been divided to eight assemblages (A-H) from which A and B have the most important zoonotic potential. All remaining genotypes have a strong commitment to various host animals. We present here the first clinical case of a human infection with the dog-specific genotype C of G. duodenalis in Slovakia. The patient, 44-year-old woman, suffered from long-term diarrhoea, abdominal pain, anorexia, weight loss, severe itching and dermatitis in the perianal area. The initial microscopic diagnosis was completed by a nested polymerase chain reaction (PCR) which revealed the first evidence of human giardiasis caused by the dog-specific genotype of G. duodenalis on a European scale. A possible role of dogs in zoonotic transmission of giardiasis and its epidemiological and public health relevance is accentuated.

Changes of the immunological and haematological parameters in rabbits after bendiocarbamate application.

The effect of bendiocarbamate application (5 mg/kg b.w.) on the haematological and immunological parameters in rabbits was evaluated. Total leukocyte cell count, erythrocyte cell count, differential cell count were determined during the period of three months of bendiocarbamate application and compared with those in healthy animals. The immunotoxic effect was evaluated by the test of ingestion ability of phagocytes (phagocytic activity and index of phagocytic activity) and proliferation activity of lymphocytes after mitogen stimulation. The significant decrease of total leukocytes, lymphocytosis and neutropaenia were found after bendiocarbamate application. The functional activities of phagocytes (expressed as phagocytic activity) and lymphocytes (proliferative activity) were significantly suppressed in rabbits treated with bendiocarbamate compared with those in control groups and values before the experiment.

Immunomodulatory effect of glucan on specific and nonspecific immunity after vaccination in puppies.

The objective of the study was to determine the immunostimulatory effect of ß-(1,3/1,6)-D-glucan in puppies. The effect exerted on the efficacy of vaccination, especially against canine parvovirus and rabies infection, was studied. The application of vaccine and glucan leads to significant increases in the nonspecific immunological parameters (phagocytic ability of leukocytes, blastogenic response of lymphocytes, metabolic and chemotactic activity of polymorphonuclear cells). The level of antibodies against canine parvovirus (Ab CPV) and rabies infection reached the most statistically significant values on the 28th day after the application of vaccine and a syrup containing ß-(1,3/1,6)-D-glucan (Group GV) as compared to the control group (Group V, puppies receiving only vaccine). Dogs without glucan supplementation did not produce such significant levels of antibodies. We can conclude that glucan has relevant immunostimulatory effects in dogs with altered immunity. The glucan product tested in this study (PleraSAN V, PLEURAN, Bratislava, Slovakia) could be used in the small animal clinical practice.

Effect of ovariohysterectomy on canine postsurgical leukocyte function.

The effect of surgery on phagocytic activity of blood leukocytes and mitogen-induced blastogenesis of lymphocytes was studied in fourteen dogs. Simple ovariohysterectomy with anaesthesia induced by ketamine and xylazine or by ketamine, xylazine and halothane caused a short nonsignificant depression of phagocytic activity that persisted for four hours after surgery. Ingestion capacity of leukocytes decreased significantly immediately after surgery. Mitogen-induced blastogenesis of lymphocytes was depressed significantly in the first 48 hours and despite partial recovery this parameter did not reach the value of the control groups until the end of observation (7 days). A more conspicuous decrease of blastogenic response of blood lymphocytes to mitogens was found after the use of ketamine and xylazine in a dose maintaining anaesthesia. Anaesthesia with ketamine and xylazine in the lower dose and maintained with halothane resulted in a later improvement of the blastogenic response of lymphocytes.

A survey for BVDV antibodies in cattle farms in Slovakia and genetic typing of BVDV isolates from imported animals.

A serological survey for bovine viral diarrhoea virus (BVDV) antibodies on a collection of 1295 serum samples obtained from 6-12 months old cattle originating from 45 farms in Slovakia was carried out. On 13 farms more than 90% of the examined animals were seropositive, on 14 farms 71-90% seroprevalence was observed, on 13 farms only 50-70% animals were found to be positive for BVDV antibodies, while the remaining 5 farms showed fewer than 50% seropositive animals. The average incidence of BVDV antibodies (around 70%) was similar as determined 30 years ago. Of 84 serum samples from seronegative animals originating from 14 farms in which 70-98% seropositivity was observed, six were positive in Ag-BVDV ELISA indicating persistently infected (PI) cattle. On a farm to which animals were imported from abroad, a BVD outbreak was observed. Of 110 animals tested, four were positive in Ag-ELISA indicating the presence of PI cattle on this farm. Genetic typing of two isolates from imported animals performed by RT-PCR (324/326 primers from 5'-UTR), sequencing of PCR products and computer-assisted phylogenetic analysis revealed that they belong to BVDV-1 h group.