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The impact of in vitro isolation and molecular characterisation of Neospora caninum as well as sequence analyses was studied. The brain homogenate of a naturally Neospora-infected dairy cow (positive in ELISA and Western blot) was intraperitoneally inoculated into Mongolian gerbils (Meriones unguiculatus). The brain of gerbils on day 60 post-inoculation was homogenized, and, after trypsin-digestion, cultured on Vero cells. Neospora-like tachyzoites were first observed after 77 days of cultivation. The parasite was confirmed by polymerase chain reaction (PCR) using Neospora-specific primers Np21 and Np6. The PCR product of the first Slovak isolate (NC-SKB1) was subsequently sequenced and published in GenBank under accession number GU300774. Sequencing and BLAST search identified the isolate as N. caninum.
Parasitic diseases of livestock together with poor welfare conditions can negatively affect the health status and production of small ruminants. Protozoan parasites and tick-borne infectious agents are common threat of livestock including small ruminants mostly during the pasture season. Therefore the priority of the study was to analyse the circulation and presence of two protozoan parasites Toxoplasma gondii and Neospora caninum as well as tick-transmitted bacterium Anaplasma phagocytophilum in one selected goat farm in Eastern Slovakia. Throughout a three-year study period we have repeatedly screened the sera and blood of goats and dogs from monitored farm. In total, 343 blood serum samples from 116 goats were examined by ELISA. The mean seropositivity for T. gondii was 56.9% (66/116, CI (95%) = 48–66.0) and 15.5% (18/116, CI (95%) = 9.3–22.7) for N. caninum. The permanent occurrence of anti-Toxoplasma and anti-Neospora antibodies was detected in repeatedly examined goats during the whole monitored period. The presence of both parasites in the flock was analysed by PCR. DNA of T. gondii was confirmed in 12 out of 25 Toxoplasma-seropositive goats and N. caninum in 14 samples out of 18 Neospora-seropositive animals; four goats were co-infected with both pathogens. The risk of endogenous transmission of both parasites was pursued by examination of 41 kid’s sera, where seropositivity for toxoplasmosis was 31.7% and for neosporosis 14.6%. In dogs 61.1% seropositivity for T. gondii and 38.9% for N. caninum was found, however, their faeces were negative for coccidian oocysts. Eight out of 108 tested animals were infected with A. phagocytophilum, the causative agent of tick-borne fever. Seven of them were simultaneously infected with T. gondii and A. phagocytophilum, out of which four goats were concurrently infected with all three pathogens.
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