The impact of climatic conditions on the dynamics of tick-borne encephalitis in Slovakia in 2012-2016.

OBJECTIVES: The main aim of our work was to analyse the development of the dynamics of tick-borne encephalitis (TBE) in connection with climatic conditions in Slovakia in 2012-2016. MATERIAL AND METHODS: We performed the analysis based on the data provided by the Epidemiological Information System and the Slovak Hydrometeorological Institute. The study group consisted of 639 patients with confirmed diagnosis. RESULTS: The highest incidence of TBE was recorded in 2016. The highest standardized incidence rate of TBE was in the districts of the Trencín, Zilina and Banská Bystrica regions. The relation of TBE to air temperature showed that most cases of TBE were recorded at an air temperature of 10-20 °C during the months of May to October in 2012-2016. The relationship between air temperature and number of days with snow cover and the number of TBE cases proved to be statistically significant (p-value < 0.001). There is a statistically significant difference in the average number of disease cases according to the air temperature category (p-value = 0.03). This disease occurs mainly in districts with an altitude of 200-400 m a. s. l. The dynamics of TBE in Slovakia is two-peaked with a decline in August. The main season of the disease lasts from May to October, peaking during the summer months of June and July. CONCLUSION: The results of the study point to a prognosis of the development of the disease in connection with air temperature. Based on the findings that in recent years we have observed a slightly increasing trend of TBE in Slovakia due to climate change, this disease is considered a persistent public health problem.

Micromeritic procedures in assessing antinuclear antibody patterns in immunofluorescent assay.

AIM: The aim of this study was to introduce a micromeritic procedure (a statistical approach for small objects) in indirect immunofluorescence assay (IFA) to find objective quantitative parameters of antinuclear antibody (ANA) patterns which could support a diagnosis of auto-immune diseases. MATERIALS AND METHODS: Sera of patients with systemic autoimmune diseases, McCoy-Plovdiv serum-free cell line, goat anti-human immunoglobulin-G FITC-conjugate, fluorescent microscope, computer-assisted digital image processing, analysis using a micromeritic procedure, ANOVA. RESULTS: Three ANA fluorescent patterns (homogeneous, rim and speckled) were analyzed by the micromeritic procedure. Parameters for the image brightness of the pixels (pixel grey value) were obtained and discussed as objective characteristics of fluorescent patterns: maximum ANA-linkage volume and surface density were established for the objects with speckled localization pattern. CONCLUSION: The micromeritic method for getting objective quantitative values of ANA fluorescent patterns in indirect immunofluorescent assay might be a valuable tool aiding in immunological diagnosing if integrated in a laboratory software package.

Phylogenetic relationship of Hepatozoon blood parasites found in snakes from Africa, America and Asia.

The blood parasites from the genus Hepatozoon Miller, 1908 (Apicomplexa: Adeleida: Hepatozoidae) represent the most common intracellular protozoan parasites found in snakes. In the present study, we examined 209 individuals of snakes, from different zoogeographical regions (Africa, America, Asia and Europe), for the occurrence of blood parasites using both molecular and microscopic examination methods, and assess phylogenetic relationships of all Hepatozoon parasites from snakes for the first time. In total, 178 blood smears obtained from 209 individuals, representing 40 species, were examined, from which Hepatozoon unicellular parasites were found in 26 samples (14·6% prevalence). Out of 180 samples tested by molecular method polymerase chain reaction (PCR), the presence of parasites was observed in 21 individuals (prevalence 11·6%): 14 snakes from Africa belonging to six genera (Dendroaspis, Dispholidus, Mehelya, Naja, Philothamnus and Python), five snakes from Asia from the genus Morelia and two snakes from America, from two genera (Coluber and Corallus). The intensity of infection varied from one to 1433 infected cells per 10 000 erythrocytes. Results of phylogenetic analyses (Bayesian and Maximum Likelihood) revealed the existence of five haplotypes divided into four main lineages. The present data also indicate neither geographical pattern of studied Hepatozoon sp., nor congruency in the host association.

Serological and molecular detection of Borrelia burgdorferi sensu lato and Anaplasmataceae in rodents.

The presence of Anaplasma spp. and Borrelia burgdorferi sensu lato in rodents from Eastern Slovakia were followed by serological and molecular methods. The seroprevalence for Borrelia was detected in 16.6 %, for Anaplasmataceae (APT) in 13.2 % and co-occurrence of Borrelia and APT in 7.5 %. Out of 110 ear biopsies of rodents, 5 were B. afzelii-positive. Five biopsies tested positive with the Ehr521-Ehr747 primers amplifying all the members of the family APT. A. phagocytophilum was detected in 1.8 %, 2.7 % were infected with Anaplasma-like organisms. Co-occurrence of Borrelia and Anaplasma in ear biopsies was found in 1.8 %. The circulation of both Borrelia and Anaplasma in the region of Eastern Slovakia was confirmed.

Potential foci of Lyme borreliosis in towns.

In 1997 the presence of ticks Ixodes ricinus infected with the agent of Lyme borreliosis was detected on the vegetation of Kosice urban agglomeration. The density of ticks on the vegetation of pericentral area islands of forest stands and neglected gardens was 2.4 times lower than that observed in a peripheral park forest. Only sporadical findings of ticks were reported from the industrial central part of town with remnants of uncultivated vegetation. Central parks and housing estates were actually tick-free. Borrelia-infected ticks, from 12.8 to 15% were found in the park forest and pericentral areas. The results indicate potential risks for humans and domestic animals coming in contact with borrelia-infected ticks, also in the territory of large towns.

Expression of outer surface proteins A and C of Borrelia burgdorferi in Ixodes ricinus.

A total of 472 field-collected Ixodes ricinus ticks from southern Germany were investigated by immunofluorescence for the presence of Borrelia burgdorferi with a polyvalent rabbit immune serum and with monoclonal antibodies specific for outer surface proteins A and C (OspA and OspC, respectively). Borreliae were detected in 90 ticks with the polyvalent immunofluorescence assay. Infection rates in adults (females, 20.2%; males, 25.2%) were significantly higher than in nymphs (12.1%). OspA was detected in 77 ticks and OspC was detected in only 1 tick with the respective monoclonal antibodies. We therefore conclude that B. burgdorferi in unfed I. ricinus ticks usually expresses OspA and very rarely OspC.

Role of small mammals in the epidemiology of toxocariasis.

Studies were conducted on the role of small mammals in maintaining toxocariasis foci in urban, rural and montane biotopes. The lowest relative density of small mammals was recorded in the urban locality and the highest in the rural and montane localities. Anti-Toxocara antibodies were most frequently detected in synanthropic and hemisynanthropic species Mus musculus, Apodemus agrarius and Micromys minutus--32.0, 30.4 and 25.0%, respectively. The highest seropositivity was found in small mammals from the urban and rural localities--22.2 and 21.6%, respectively. Toxocara canis was most prevalent in urban stray dogs (75.0%) and least prevalent in foxes from the montane locality (7.0%). The prevalence of Toxocara cati in cats at the urban, rural and montane localities was 66.2, 65.2 and 76.9%, respectively. In clinically healthy human populations, the highest seroprevalence was detected in the rural locality (14.0%). Children of the same area were 3 times more seropositive (12.9%) than those from the urban and montane localities (4.3 and 4.0%). Our studies suggest an important role for small mammals as paratenic hosts--reservoirs of Toxocara larvae--in maintaining toxocariasis foci. In this respect toxocariasis may be classified as an anthropopurgic focal zoonosis.

[Examination of ticks for the presence of Borrelia sp. in Kosice and the surrounding region. Preliminary results]. / Vysetrenie kliestov na prítomnost Borrelia sp. v Kosiciach a okolí. Predbezné výsledky.

The authors present preliminary data on the infestation of ticks Ixodes ricinus with Borrelia in the town of Kosice and surroundings. The authors found a mean 4.7% positivity which means that more systematic attention should be paid to the problem.