P. Dubinský

Cystic echinococcosis in Turkey: genetic variability and first record of the pig strain (G7) in the country

A sample of 22 Echinococcus granulosus isolates collected from 12 sheep and ten humans from a focus of cystic echinococcosis in western Turkey was examined by DNA sequencing of four mitochondrial genes (cox1, atp6, nad1, rrnS). Results demonstrated the presence of two species of E. granulosus complex, E. granulosus sensu stricto and E. canadensis. Of E. granulosus sensu stricto, the G1 genotype (including three microvariants) was found in 17 isolates from humans and sheep, the G3 genotype and an intermediate form G1/G3 in one isolate each (both from sheep). Of E. canadensis, the pig strain G7 was found in three isolates from sheep and human. This is the first report of this strain in Turkey. Its presence has implications for local control programs due to its shorter maturation rate in dogs compared with E. granulosus sensu stricto. Goat and/or wild boar are likely reservoirs for G7 in the region. We provided further data on the pattern and frequency of nucleotide substitutions within the G1/G3 cluster. Based on our results and GenBank records, G2 (Tasmanian sheep strain) is not considered as a discrete genotypic unit, as its sequences at polymorphic sites conform to microvariants of both G1 and (more often) G3.

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.

Effect of various doses of infectiveToxocara canis andToxocara cati eggs on the humoral response and distribution of larvae in mice

The effect of 5–2,500 infectiveToxocara canis and 5–1,000T. cati eggs on the humoral immune response and on the distribution of larvae in the organism was studied in paratenic hosts — inbred C57BL6/J mice. With each dose ofT. canis eggs the maximal antibody level was recorded on day 56 post infection and was followed by a moderate decline that lasted until day 154 of the experiment. A correlation between the antibody level and the egg count was observed only with the infective dose of 5–50 eggs. A more rapid occurrence of antibodies was recorded in mice infected with a high dose of eggs. In those given 5 and 7T. cati eggs the antibody level exceeded the extinction threshold value only from day 21 to day 84. Low doses ofT. canis (n=5) andT. cati (n=7) eggs caused a comparable distribution of larvae in mice, and the larval recoveries on day 70 post infection ranged between 10.00% and 25.74%. Following a dose of 500T. cati eggs, 22.28% of the larvae were recovered, although only 1.08% were localized in the brain. A dose of 1,000T. canis eggs yielded, 36.37% of the larvae, with as much as 28.13% being found in the brain.

Morphological and genetic characterization of Echinococcus granulosus in the Slovak Republic

The present study was undertaken to characterize more thoroughly aetiological agent of cystic echinococcosis that still represents a health problem in the Slovak Republic. Sequencing of the mitochondrial ND1 gene, Random Amplified Polymorphic DNA (RAPD) analysis and isoenzyme analyses were used to characterize genetically protoscolices of 37 isolates from pigs and cattle and two isolates from humans collected in different regions of the Slovak Republic. Solely the G7 genotype (pig strain) was identified by ND1 sequences in all 14 pigs isolate examined by this method. This genotype was also found in two human patients and in two cattle. The exclusive finding of the G7 variant has implications for the implementation of control strategies given its unique developmental and biological properties. Despite striking morphological variability, a low degree of the intra-strain variability was found in the Slovak Echinococcus granulosus isolates with the genetic methods used. This was particularly associated with the shape and size of hooks, number of testes and shape of ovaries and uterus. GPI and MDH enzymes were found suitable for the discrimination of G7 and G1 (=cosmopolitan sheep strain) variant.

Dirofilariosis in Slovakia - a new endemic area in Central Europe

SummaryThe first systematic research on canine dirofilariosis in Slovakia started in February 2007. In total 287 dogs of various age, sex, breeds, and utilization from southern Slovakia were examined within the study until September 2007. Microfilariaemia was detected in 99 (34.5 %) blood samples. Histochemical staining and PCR approach were used for Dirofilaria species identification. Dirofilaria repens was confirmed in all infected dogs; it occurred in 92 (32.1 %) dogs as a single infection and in 6 (2.1 %) dogs in mixed infection with Dirofilaria immitis. For the first time both species were detected in Eastern Slovakia. Outdoor keeping and age higher 3 years become evident as important risk factors. Male individuals and dogs with long hair were significantly more frequently infected when compared with females and short hair breeds.The present study revealed the occurrence of highly endemic area of dirofilariosis in Slovakia and certified that climatic conditions in the country are favourable for development of the parasite and for further spread of this zoonotic infection.

Dirofilaria infections in working dogs in Slovakia.

A monitoring programme aimed at the diagnosis of subcutaneous dirofilariasis and heartworm disease in working (police and military) dogs in Slovakia has been performed during the period of September 2007 to February 2008. In co-operation with the Ministry of the Interior and the Ministry of Defence, in total, 710 dogs (591 police dogs and 119 military dogs) were investigated for the presence of microfilariae in blood. All police and military dogs in active service held on the territory of Slovakia were included. Microfilariae were detected in 118 (20.0%) police dogs and 10 (8.4%) military dogs. The most infected individuals originated from southern parts of Slovakia (Trnava region 53.6% and Nitra region 39.6%); the prevalence was low in northern regions (Zilina 3.1% and Presov 6.6%). In several districts of southern Slovakia, the prevalence of subcutaneous dirofilariasis in working dogs exceeded 40%. In all infected animals, the autochthonous origin of the disease was confirmed; however, due to the frequent movement of working dogs, it was not possible to identify the exact locality of infection. At present, a dog living in Nemsová village in Trencín district (north-western part of the country) is regarded as the northernmost localized autochthonous case of subcutaneous dirofilariasis in Slovakia. In three dogs, co-infection of Dirofilaria repens and Dirofilaria immitis was detected. High prevalence rates in working dogs and the zoonotic characteristic of the disease represent an undoubtedly important veterinary and medical problem that requires the urgent introduction of prophylactic and control measures.

Endoparasites of red fox (Vulpes vulpes) in the Slovak Republic with the emphasis on zoonotic species Echinococcus multilocularis and Trichinella spp.

SummaryDue to specific geographical localization, climatic and geomorphologic conditions, several serious parasitic diseases circulate in the territory of the Slovak Republic that makes this area an ideal model territory of the central European red fox system. The red fox is an important reservoir host of parasites, which can be spread to another animals and humans. Our study was aimed at determining the current prevalence of certain parasites in red foxes from the entire territory of the Slovak Republic and identifies some ecological factors influencing their epidemiology. Within the first systematic investigation of red foxes carried out between the years 2000 and 2006 in total 4026 foxes were examined for Echinococcus multilocularis (prevalence 31.1 %) and 4699 foxes were investigated for the presence of Trichinella spp. larvae (10.4 % infected). The results of the next separate study revealed that 83.3 % of 1198 red foxes in the Slovak Republic had coccidian oocysts and helminth eggs in their faeces. Fifteen helminth species including two trematode, four cestode and nine nematode species were detected by coprological examination. Nine of these parasite taxa have zoonotic potential: Capillaria spp. (prevalence 22.4 %), Ancylostoma caninum (18.1 %), Toxocara canis (12.5 %), Taenia spp. (12.2 %), Mesocestoides spp. (5.8 %), Strongyloides stercoralis (1.6 %), Hymenolepis diminuta (0.6 %), Dipylidium caninum (0.4 %) and Opisthorchis felineus (0.3 %). Toxascaris leonina was the most common helminth species found in this survey (42.9 %).

A seroepidemiological study of human Toxocara infection in the Slovak Republic

Sera of 908 blood donors, average age 35 years, living in towns and villages of the Slovak Republic were tested for Toxocara canis antibodies using an ELISA-IgG test. The seroprevalence in the healthy population was estimated at 13.65%. Antibodies were predominantly demonstrated in the sera from persons living in rural communities (17.09%) compared to the sera investigated from urban communities (11.82%). Women had a significantly higher seropositivity than men. The serological response in 2703 sera from patients suspected of toxocarosis was compared with their clinical manifestation, sex and age. Toxocara antibodies were found in 27.41% of suspected patients. Although the seroprevalence in adults was significantly higher among women than men, in children, boys had a higher seroprevalence than girls. The main clinical manifestations were leucocytosis and eosinophilia (46%), ocular disorders (36%), lung, liver and neurological disorders (26%). The most severe forms of this disease were observed in 2 to 5-year-old children with geophagy.

The first finding of Neospora caninum and the occurrence of other abortifacient agents in sheep in Slovakia

Neosporosis is an infection of animals caused by an intracellular coccidian parasite, Neospora caninum, closely related to Toxoplasma gondii. The parasite is one of important abortifacient agents of bovine abortions worldwide. The aim of the study was to detect the prevalence of anti-Neospora antibodies in dairy aborting sheep from two eastern Slovak regions and to compare it with the occurrence of other potential abortifacient agents. Sera of 382 sheep, mainly the Improved Valachian and Merino breed, were tested for the presence of anti-Neospora and anti-Toxoplasma antibodies by ELISA, anti-Leptospira sp. by micro-agglutination-assay and anti-Chlamydophila antibodies using the complement fixation test. The mean seroprevalence of N. caninum was 3.7% and of T. gondii, 24.3%. This phenomenon of higher susceptibility of sheep to T. gondii is in the opposite of N. caninum infection in cattle. Anti-Leptospira antibodies were observed in 2.9% of serum samples with titres from 800 to 1600, whereas IgG antibodies against Chlamydophila abortus were found in 13.6% with titres from 64 to 1024. Half of N. caninum positive sera were simultaneously positive for T. gondii and one sample for C. abortus. From examined abortifacient agents the most important, from the frequency point of view, were toxoplasmosis (24.3%) and chlamydiosis (13.6%). No significant association between the frequencies of the abortions and mean seroprevalence of the abortifacient agents in Kosice region was determined. Likewise, no significant differences between the mean seroprevalence of neosporosis and toxoplasmosis in the two regions were detected. The first survey of neosporosis in aborting sheep from eastern Slovakia revealed a low prevalence resulting in a lower impact on reproduction losses in these small ruminants.

Long-term survey on Trichinella prevalence in wildlife of Slovakia.

In Slovakia, monitoring the prevalence of Trichinella spp. in wildlife was performed since 2000 in the main reservoir animals, the red fox (Vulpes vulpes) and wild boar (Sus scrofa), using artificial digestion method as recommended by International Commission on Trichinellosis. The results of investigation performed in 5270 red foxes showed that Trichinella infection is widespread across Slovakia and prevalence increased significantly from 4.9% in 2000 to 20.5% in 2007. Recently, a higher Trichinella prevalence (0.11%) in wild boars was also demonstrated. The results indicate that foxes and wild boars are involved in the spread of Trichinella, although the latter host species seems to play a secondary role in the maintenance of the sylvatic cycle in Slovakia. Trichinella britovi is the predominant species circulating in Slovakia, both in foxes and wild boars, and Trichinella spiralis occurs only sporadically. Mixed infections of T. britovi and Trichinella pseudospiralis were recorded in 2005 in one wild boar from Eastern Slovakia and in 2006 in one red fox from the same region. These findings are important with respect to an outbreak caused by T. pseudospiralis in a pig farm in the same district 3 years ago. This study provides a complex picture on Trichinella occurrence in all regions of Slovakia and may be a good basis for evaluating the risk of parasite transmission to the domestic cycle and human beings.