Diverse tick-borne microorganisms identified in free-living ungulates in Slovakia

Background: Free-living ungulates are hosts of ixodid ticks and reservoirs of tick-borne microorganisms in central Europe and many regions around the world. Tissue samples and engorged ticks were obtained from roe deer, red deer, fallow deer, mouflon, and wild boar hunted in deciduous forests of south-western Slovakia. DNA isolated from these samples was screened for the presence of tick-borne microorganisms by PCR-based methods. Results: Ticks were found to infest all examined ungulate species. The principal infesting tick was Ixodes ricinus, identified on 90.4% of wildlife, and included all developmental stages. Larvae and nymphs of Haemaphysalis concinna were feeding on 9.6% of wildlife. Two specimens of Dermacentor reticulatus were also identified. Ungulates were positive for A. phagocytophilum and Theileria spp. Anaplasma phagocytophilum was found to infect 96.1% of cervids, 88.9% of mouflon, and 28.2% of wild boar, whereas Theileria spp. was detected only in cervids (94.6%). Importantly, a high rate of cervids (89%) showed mixed infections with both these microorganisms. In addition to A. phagocytophilum and Theileria spp., Rickettsia helvetica, R. monacensis, unidentified Rickettsia sp., Coxiella burnetii, "Candidatus Neoehrlichia mikurensis", Borrelia burgdorferi (s.l.) and Babesia venatorum were identified in engorged I. ricinus. Furthermore, A. phagocytophilum, Babesia spp. and Theileria spp. were detected in engorged H. concinna. Analysis of 16S rRNA and groEL gene sequences revealed the presence of five and two A. phagocytophilum variants, respectively, among which sequences identified in wild boar showed identity to the sequence of the causative agent of human granulocytic anaplasmosis (HGA). Phylogenetic analysis of Theileria 18S rRNA gene sequences amplified from cervids and engorged I. ricinus ticks segregated jointly with sequences of T. capreoli isolates into a moderately supported monophyletic clade. Conclusions: The findings indicate that free-living ungulates are reservoirs for A. phagocytophilum and Theileria spp. and engorged ixodid ticks attached to ungulates are good sentinels for the presence of agents of public and veterinary concern. Further analyses of the A. phagocytophilum genetic variants and Theileria species and their associations with vector ticks and free-living ungulates are required.Fil: Kazimírová, Mária. Slovak Academy of Sciences. Institute of Zoology; EslovaquiaFil: Hamšíková, Zuzana. Slovak Academy of Sciences. Institute of Zoology; EslovaquiaFil: Spitalská, Eva. Slovak Academy of Sciences. Institute of Virology. Biomedical Research Center,; EslovaquiaFil: Minichová, Lenka. Slovak Academy of Sciences. Institute of Virology. Biomedical Research Center,; EslovaquiaFil: Mahríková, Lenka. Slovak Academy of Sciences. Institute of Zoology; EslovaquiaFil: Caban, Radoslav. Široká ; EslovaquiaFil: Sprong, Hein. National Institute for Public Health and Environment.Laboratory for Zoonoses and Environmental Microbiology; Países BajosFil: Fonville, Manoj. National Institute for Public Health and Environment.Laboratory for Zoonoses and Environmental Microbiology; Países BajosFil: Schnittger, Leonhard. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Kocianová, Elena. Slovak Academy of Sciences. Institute of Virology. Biomedical Research Center,; Eslovaqui

ELEVATED TRANS-MAMMARY TRANSMISSION OF Toxocara canis LARVAE IN BALB/c MICE

Toxocariasis is a widespread zoonosis and is considered an important worldwide public health problem. The aim of this study was to investigate the frequency of trans-mammary Toxocara canis infection in newborn BALB/c mice nursed by females experimentally infected with 1,200 eggs after delivery. After 50 days of age, the presence of larvae in different organs of the offspring was investigated. Trans-mammary infection was confirmed in 73.9% of the mice that had been nursed by infected females. These data show a high trans-mammary transmission of T. canis and confirm the significance of this transmission route in paratenic hosts

EFFICACY OF NITAZOXANIDE AGAINST Toxocara canis: LARVAL RECOVERY AND HUMORAL IMMUNE RESPONSE IN EXPERIMENTALLY INFECTED MICE

SUMMARY The efficacy of nitazoxanide (NTZ) against toxocariasis was investigated in an experimental murine model and results were compared to those obtained using mebendazole. Sixty male BALB/c mice, aged six to eight weeks-old, were divided into groups of 10 each; fifty were orally infected with 300 larvaed eggs of T. canisand grouped as follows, G I: infected untreated mice; G II: infected mice treated with MBZ (15 mg/kg/day) 10 days postinfection (dpi); G III: infected mice treated with NTZ (20 mg/kg/day) 10 dpi; G IV: infected mice treated with MBZ 60 dpi; G V: infected mice treated with NTZ 60 dpi; GVI: control group comprising uninfected mice. Mice were bled via retro-orbital plexus on four occasions between 30 and 120 dpi. Sera were processed using the ELISA technique to detect IgG anti- Toxocaraantibodies. At 120 dpi, mice were sacrificed for larval recovery in the CNS, liver, lungs, kidneys, eyes and carcass. Results showed similar levels of anti- ToxocaraIgG antibodies among mice infected but not submitted to treatment and groups treated with MBZ or NTZ, 10 and 60 dpi. Larval recovery showed similar values in groups treated with NTZ and MBZ 10 dpi. MBZ showed better efficacy 60 dpi, with a 72.6% reduction in the parasite load compared with NTZ, which showed only 46.5% reduction. We conclude that administration of these anthelmintics did not modify the humoral response in experimental infection by T. canis. No parasitological cure was observed with either drug; however, a greater reduction in parasite load was achieved following treatment with MBZ

Serological and Molecular Survey of Serious Intracellular Coccidia in Red Foxes in Eastern Slovakia

We tested the sera or meat juices of 215 red foxes (Vulpes vulpes), collected during 2009-2014 at different localities of Eastern Slovakia for the presence of anti- Toxoplasma and anti-Neospora antibodies. We also examined the brains or muscle tissues and uncoagulated blood samples for the presence of both parasite’s DNA. The mean seropositivity to T. gondii was 72.6% (95% Confidence Interval CI 66.1-78.4) and to N. caninum 33.9% (95% CI 27.7-40.7). We observed higher Toxoplasmaseropositivity in adults (71.1%) than in juveniles (60.0%). Neospora-seropositivity was almost the same in both age groups about of 30%. In general, we observed less frequently the DNA of parasites, T. gondii (14.5%) and N. caninum (20.3%) in the tissue samples and uncoagulated blood samples. Coccidioses are considerably common in red foxes and circulate in locations of Eastern Slovakia. The high infection rate in foxes is probably due to their infected prey. On the other hand, the contamination of the environment with oocysts and their subsequent transfer to other farm and wild animals is also possible

Trichinellosis in the Slovak Republic

The occurrence of trichinellosis in farm or wildlife animals in some areas pose the permanent risk of infection in humans. In Slovakia, where only the sylvatic trichinellosis exists, the humans trichinellosis is sporadic. The most common cause of infection is wild boar meat. Unusual local eating customs may also favour an outbreak of disease in humans. The outbreak of trichinellosis in Slovakia in 1998, when 336 people were affected, was of the same kind. Trichinella britovi was the agent of the infection

Congenital trichinellosis ? Case report

A large trichinellosis outbreak in the Slovak Republic caused by the species Trichinella britovi and affecting 336 people also affected a pregnant woman. The mother was infected in the 10thth week of pregnancy and was treated with mebendazole. On her own request abortion was performed in the 22nd week of pregnancy. Medium lgM and high lgG anti-Trichinella antibody titres were found. The placenta, body cavities liquid, tissues and organs of the foetus contained 0.02 - 30 larvae per gram of tissue, measuring 0.68 ± 0.05 - 1.17 ± 0.07 mm, with blurred inner structure. Immunocytochemical examination identified Trichinella larvae that infected the foetus in the early stage of development

Congenital trichinellosis ? Case report

A large trichinellosis outbreak in the Slovak Republic caused by the species Trichinella britovi and affecting 336 people also affected a pregnant woman. The mother was infected in the 10thth week of pregnancy and was treated with mebendazole. On her own request abortion was performed in the 22nd week of pregnancy. Medium lgM and high lgG anti-Trichinella antibody titres were found. The placenta, body cavities liquid, tissues and organs of the foetus contained 0.02 - 30 larvae per gram of tissue, measuring 0.68 ± 0.05 - 1.17 ± 0.07 mm, with blurred inner structure. Immunocytochemical examination identified Trichinella larvae that infected the foetus in the early stage of development