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The protozoan Toxoplasma gondii is an obligate intracellular parasite that infects a wide range of warm-blooded vertebrates. The data about the occurrence of toxoplasmosis in slaughter pigs in the Slovak Republic are still missing. The aim of our study was to estimate the prevalence of toxoplasmosis in pigs from Slovakia during the period of 2006–2010 by ELISA and PCR methods. In sera of 970 slaughter pigs, 2.16% seropositivity to T. gondii was detected. In tissue samples of seropositive pigs the presence of T. gondii DNA was confirmed. In six monitored Slovak regions the seropositivity varied between 1.11 and 3.48%. The statistically significant differences were recorded between the Košice and Prešov region. The seroprevalence of toxoplasmosis in sows (4.26%) was two times higher than that in slaughter pigs (2.06%) (OR = 2.12; 95% CI = 0.48–9.36). Presence of Toxoplasma gondii in tissues of seropositive pig isolates was confirmed by TGR1E and B1 genes and analysis of DNA polymorphism at SAG2 and ROP1 genes revealed the presence of virulent strain of genotype I in 85.7% of infected pigs and an avirulent strain (genotype II) in 14.3% of pigs.
The course of anti-Neospora antibody kinetics during two consecutive pregnancies has been evaluated in five chronically infected dairy cows. The blood samples of cows were collected monthly. Anti-Neospora antibodies in blood sera were detected by indirect ELISA (ID-VET, France). During whole period of the study cows remained seropositive; with S/P% values (iELISA) ranging from 94 to 214%. The antibody kinetics determined by iELISA showed a significant increase (P<0.0059) of specific IgG antibodies in the third trimester of both pregnancies. The monitored cows gave birth to 10 healthy calves (4 steers and 6 heifers). To confirm the occurrence of vertical transmission of Neospora from mothers to offspring in the herd, dam-daughter serology was performed. Anti-Neospora antibodies was found in 4 from 6 heifers (>6 months old). Study presents original data reporting on the very similar persistent pattern of anti-Neospora antibody levels during the third trimester of pregnancies in all five dairies. Based on the high seropositivity of female offspring, the reactivation of a latent infection of cows rather than a re-infection can be supposed.
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.
During the period of 2000-2004, 3,096 red foxes from the whole territory of the Slovak Republic were sampled and examined parasitologically for infections with Echinococcus multilocularis, causative agent of serious alveolar echinococcosis in humans. Relations between prevalence of the parasite in individual regions of Slovakia and some environmental factors were weighted. During the study period, great differences of prevalence and infection intensity were found on a regional level and significant between-year fluctuation of both parameters was observed. High-endemic foci with an estimated prevalence of more than 30% were detected in the northern and central part of the country. Climatic conditions, including low mean annual air temperature, high mean annual rainfall and the high humidity of the soil, showed to be important for E. multilocularis distribution. Significant correlation was calculated between prevalence of the tapeworm, mean annual precipitation values, and population density of small mammals.
Toxoplasma gondii is a protozoan parasite of great medical and veterinary importance. The aim of this study was to determine the seroprevalence of toxoplasmosis in wild boars hunted in the Slovak Republic in 2003. Examination of 320 wild boars revealed a seroprevalence of 8.1%. The majority of seropositive wild boars came from the north-western and southern regions of Slovakia. This study indicates that T. gondii infection is common in wild boars in the Slovak Republic, underlines its zoonotic potential and the importance of high standards of hygiene during the handling of game.
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.
The extensive distribution of Echinococcus multilocularis cestode from endemic alpine areas to the parts of Central Europe has been recorded in recent years. The first confirmed finding of E. multilocularis in Slovakia was recorded in 1999 in the area adjacent to the Polish border. At present, this serious zoonosis occurs almost across the whole territory of Slovakia. The occurrence of these tapeworms in red foxes (Vulpes vulpes) at the border regions of Slovakia and Poland has been monitored. In these districts, out of 152 faecal samples examined, 36.2% were positive for the coproantigen-ELISA. With the sedimentation and counting technique the prevalence of E. multilocularis in red foxes was up to 38.8%. The examination of foxes from neighbouring districts revealed worm burden ranging from 1–15,000 specimens, but the majority of animals harboured medium number of tapeworms. In the Small Carpathian and Sub-Carpathian regions of Poland, out of 65 samples examined, 13.8% were coproantigen positive. Using the small intestine scraping method only 6.1% prevalence of E. multilocularis in red foxes was determined, mostly with a high worm burdens over 1,000 specimens. The results suggest possible transborder transmission of E. multilocularis, the causative agent of serious alveolar echinococcosis.
Sixty-eight dogs were examined for the presence of Encephalitozoon spp. antibodies. Twenty-one dogs (30.9%) were healthy without any clinical signs of diseases. Forty-seven animals (69.1%) developed clinical symptoms of diseases such as chronic otitis externa, conjunctivitis, upper respiratory tract inflammation, status epilepticus, pyodermatitis, skin hypersensitivity, demodicosis, flea allergy. Different detection methods of encephalitozoonosis including IFAT (Indirect Immunofluorescence Antibody Assay), in vitro cultivation, SDS PAGE electrophoresis, Western blot and PCR were applied. There were 33 (48.5%) positive reacting sera to E. cuniculi II (mouse type) antigen using IFAT, including 9 positive samples obtained from clinically healthy dogs. Sixteen samples with the antibody titer equal to 1 : 256 were then tested by Western Blot. Most of the samples reacted with E. cuniculi II and III antigens. The presence of E. intestinalis antibodies was lower and just a few samples reacted with E. hellem antigen. The electrophoretic analysis of the encephalitozoon strains used as antigens confirmed that they differ primarily in the molecular size. The strain of type II (mouse) expressed a double strip at 54 and 58 kDa level. The strain of type III (dog) expressed a wide strip at 59 kDa. E. cuniculi types II and III are more related in protein structure in comparison to the other analyzed strains. When PMP1/PMP2 primers were used in PCR, the size of the amplified product was 268 bp for E. cuniculi and 270 bp for E. intestinalis. A species-specific primer pair for E. cuniculi ECUNF/ECUNR gave a 549 bp fragment and V1/SI-500 primers specific for E. intestinalis gave a 370 bp fragment.
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