Zoonotic assemblages of Giardia duodenalis in captive non-human primates from the largest zoo in Slovakia.

Only a few studies based on multilocus characterization have been conducted on the molecular epidemiology of Giardia duodenalis in captive nonhuman primates (NHPs). The present article provides the first report on the occurrence of G. duodenalis in the ring-tailed lemur (Lemur catta) and barbary macaque (Macaca sylvanus) kept in the zoo in Kosice, Slovakia. All samples were examined by flotation technique, with total prevalence of 17.4% (4/23). The microscopically positive samples were assayed by nested PCR and consecutively sequenced at ß-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) genes. Positive isolates were recognized as assemblage B and sub-assemblage BIV and subtype WB8 were confirmed. The identification and genotyping of this parasite in Slovakia, may help to better understand the epidemiological situation in Europe about the circulation of G. duodenalis zoonotic assemblages in NHPs.

The roundworm Strongyloides stercoralis in children, dogs, and soil inside and outside a segregated settlement in Eastern Slovakia: frequent but hardly detectable parasite.

A comparative study was carried out to evaluate the Strongyloides stercoralis infections in children and dogs inside and outside the segregated settlement in Medzev, Eastern Slovakia, and a survey of the soil within the settlement was included. Applying the Koga agar plate (KAP) culture method and microscopy examination of stool samples collected from 60 Roma and 21 nonRoma children, no larvae of S. stercoralis were detected but eggs of three nematodes (Ascaris lumbricoides, Trichuris trichiura, and Enterobius vermicularis) and cysts of two protozoan endoparasites (Giardia duodenalis and Cryptosporidium spp.) were often found. However, immunoenzymatic assay (ELISA) for the evidence of IgG antibodies against S. stercoralis showed 33.3% seroprevalence in Roma children and 23.8% prevalence in children from the majority population, attending the same school. Eosinophilia was regularly present in children with exclusive infection of S. stercoralis (eight cases) as well as in individuals suffering from mixed infections of S. stercoralis and some of the above listed parasites (16 cases); high eosinophil counts sometimes, but not always, occurred in parasitized children lacking S. stercoralis antibodies. A comparison of S. stercoralis in dogs from the settlement (40 dogs) and from a distant dog shelter (20 dogs) did not reveal remarkable differences: the direct microscopy of faecal samples revealed rhabditiform larvae in 13.3% of the dogs from the settlement (4/30) and in 10.0% of the dogs from the shelter (2/20). Out of blood samples collected from the second dog group, 55% of the dogs contained antibodies against S. stercoralis. In the soil collected from 14 various locations within the settlement, S. stercoralis larvae were observed in two samples (14.3%); however, 13 samples (92.9%) were positive for human or dog endoparasites of the genera Ancylostoma, Ascaris, Toxocara, Toxascaris, Trichuris, and Hymenolepis.

Giardia duodenalis and Giardia enterica in children: first evidence of assemblages A and B in Eastern Slovakia.

For an overview on the occurrence of Giardia assemblages in children in Eastern Slovakia, we examined 259 faecal samples of children from the segregated settlement in Medzev, 30 samples of children from the orphanage in Medzev and 40 samples of children with autism from the Special Elementary School in Kosice. Thirty-eight samples (14.67 %) from the segregated settlement, 19 samples (63.33 %) from the orphanage and two samples (5.0 %) from the Special Elementary School were positive for Giardia by flotation. The initial microscopic diagnostics were completed by the genotyping of the triosephosphate isomerase-gene loci (tpi genes) which revealed the existence of two Giardia assemblages in Slovak population, namely Giardia duodenalis (assemblage A) and Giardia enterica (assemblage B). These results represent the first evidence of A and B assemblages in children in Slovakia. Epidemiological significance and the impact on the public health of Giardia infection are highlighted.

Significantly higher occurrence of Cryptosporidium infection in Roma children compared with non-Roma children in Slovakia.

Cryptosporidiosis is considered to be a widespread world zoonosis. The occurrence of Cryptosporidium species was investigated in Roma children in a district of Eastern Slovakia and, at the same time, also in children of non-Roma parents. In total, 103 children (54 boys and 49 girls) between 0 and 14 years of age were involved in this study. Fifty-three were Roma children and 50 children represented a non-Roma control group. Fecal samples were examined: immunologically [enzyme-linked immunosorbent assay (ELISA) test to prove antigen in the feces] and by molecular analysis [nested polymerase chain reaction (PCR)]. After the sequencing of the PCR, the products were identified as species of Cryptosporidium muris. Based on the results, the relative risk (RR) of the Cryptosporidium infection occurrence was calculated and we came to the conclusion that the risk of Cryptosporidium infection was almost 12 times higher in the Roma children compared to the non-Roma children.

Detection of maduramycin residues in the tissues of chickens and pheasants by the screening test for antibiotic residues (STAR).

Maduramycin is a coccidiostat authorized as a feed additive in poultry. Council Directive 96/23/EC stipulates that monitoring of foods of animal origin for residues of coccidiostats is mandatory. The aim of the present study was to evaluate the STAR for the screening of maduramycin residues in the tissues of broiler chickens and pheasants. Both animal species were supplied feed medicated with Cygro l% premix according to recommendations for use (5 mg kg(-1) of complete feed). The residues were investigated for a period of 7 days: day 0 (the last day of the administration of maduramycin), days 1-5 (the days of the withdrawal period) and day 6 (the first day after elapse of the withdrawal period). According to STAR the positivity of the sample (the presence of residues of antibacterial substances) is indicated by a zone of inhibition exceeding 2 or 4 mm in width, depending on the test organism. Maduramycin residues were detected only on the plates seeded with the test organism Bacillus stearothermophilus var. calidolactis ATCC 10149. The results showed that there was higher potential for the presence of maduramycin residues in broiler chickens than in pheasants. All chicken tissues (muscle/thigh and breast/gizzard, liver, heart, kidneys, spleen, lungs) were positive for maduramycin (inhibition zones ≥4 mm) not only throughout the withdrawal period, but also even 5 days after elapse of the withdrawal period. In the case of pheasants the positive results were detected in the gizzard, liver, heart, kidneys, spleen and lungs. On day 5 of the withdrawal period no positive results were detected; however, on day 6 the heart and spleen were positive again.

Cellular immunological responses of pheasant during endogenous development of Eimeria colchici.

We examined the time course and histological localisation of the developmental stages of Eimeria colchici. The prepatent period in the caeca of pheasants was 6 days. The patent period began on day 7 post-infection (p.i.) and ended on day 11 p.i. with peak production of oocysts on days 8-9. The peripheral blood lymphocytes of pheasant chicks showed a significant increase in proliferation to E. colchici antigen from day 5 p.i., with peak on day 14 p.i. The metabolic activity (respiratory burst) of heterophils increased on days 3, 4 and 14 p.i. The total number of peripheral blood leukocytes and lymphocytes in the infected pheasant chicks had increased by day 2 p.i. and reached a maximum on day 4 of the experiment. Days 5 and 6 p.i. were characterised by a drop in the number of these cells.

Life cycle of Eimeria procera in experimentally infected grey partridges (Perdix perdix).

We examined exogenous and endogenous development of Eimeria procera in experimentally infected grey partridges (Perdix perdix). Our examination included data on morphology, localization, duration of schizogony and gametogony and morphology of sporulated oocysts. The endogenous stages of E. procera developed in large numbers within the epithelial cells of caecal crypts. The asexual development comprised three generations of schizonts. The first fully developed macrogametes and microgamonts were observed on Day 5 post-infection (p.i.) in histologic section. The patent period began on Day 6 p.i. and ended on Day 11 p.i. with peak production of oocysts on Days 7 and 8. Long oval oocysts of E. procera measured 25.78-28.13 microm in length and 14.06-15.24 microm in width, sporulation time ranged from 18 to 24h at 25 degrees C and from 36 to 48h at 20 degrees C.

Course of the endogenous developmental phase of selected Eimeria species in pheasants.

The localization and duration of developmental stages of Eimeria colchici and Eimeria duodenalis were studied histologically. The prepatent period of the most pathogenic species from the caeca of pheasants--Eimeria colchici--was 6 days. The patent period began on the 7th day and finished on the 11th day post-infection with the maximum production of oocysts on days 8-9. In the case of Eimeria duodenalis the prepatent period was shorter--4 days, and the duration of the patent period was 3-4 days without a significant increase in oocyst production.