State Veterinary Institute Prague

Prague, Czech Republic

State Veterinary Institute Prague

Prague, Czech Republic
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Pavlasek I.,State Veterinary Institute Prague | Maca O.,State Veterinary Institute Prague | Maca O.,Czech University of Life Sciences
Parasitology International | Year: 2017

Muscular sarcocystosis by Sarcocystis arctica was found for the first time in the Czech Republic, in different muscles of red fox (Vulpes vulpes). Cysts were slim, elongated, thread-like, whitish, 1–7 mm long, and 206–270 μm wide; bradyzoites were 7.9 × 2.7 μm in unstained wet mounts and 9.2 × 2.9 μm in cyst Giemsa-stained smears. The cyst wall was thin, with short villi-like protrusions, and no host response was observed in the histological sections. Examination of the distribution and intensity of sarcocysts in 17 different muscle groups revealed that the highest intensity was in the cranial tibial muscle (> 15 cysts in compressoria), followed by the diaphragm, forearm, and other groups (with intensities of 3–15 cysts in compressoria). Sarcocysts were detected in 3 out of 86 foxes. Genetic characterization at 18S rRNA, 28S rRNA, ITS1 and cox1, consistently showed that the species was identical with S. arctica. Interestingly, this protozoan was also detected as a co-infection in 3 foxes with the nematode Trichinella spp. for the first time. © 2017 Elsevier B.V.

Maca O.,State Veterinary Institute Prague | Maca O.,Czech University of Life Sciences
Systematic Parasitology | Year: 2012

Examination of faecal samples from semi-captive western Derby elands Taurotragus derbianus derbianus Gray, in the Bandia and Fathala Reserves of Senegal, revealed the presence of oöcysts of the genus Eimeria Schneider, 1875 that we considered to represent a new species, Eimeria derbiani n. sp. The new species possesses nearly ellipsoidal oöcysts (length/width ratio 1.3) with a bi-layered wall and an average size of 27.6 × 21.5 μm. E. derbiani possesses a micropyle covered by a micropylar cap and ovoidal, single-layered sporocysts with an average size of 14.9 × 7.7 μm, each with a Stieda body. Sporozoites of E. derbiani possess a large refractile body and a nucleus. Sporulation lasted for 2 days at 23°C. The new species is differentiated from the two species parasitising Taurotragus oryx Pallas, E. canna Triffitt, 1924 and E. triffittae Yakimoff, 1934. © 2012 Springer Science+Business Media B.V.

PubMed | University of Veterinary And Pharmaceutical Sciences Brno, University of Limoges, State Veterinary Institute Prague and Masaryk University
Type: | Journal: Veterinary parasitology | Year: 2016

Generally, rodents and other small mammals are considered as one of the sources of Toxoplasma gondii or Neospora caninum infection for cats and dogs as the definitive hosts of these two parasites, respectively. The aim of the study was to find out the prevalence of these two parasites in wild small mammals from the Czech Republic and to characterize T. gondii isolates by methods of molecular biology. A total of 621 wild small mammals were caught in the Czech Republic during years 2002-2014. Antibodies to T. gondii were detected by latex agglutination test in six (2.5%) of 240 small mammals (in two A. agrarius and four A. flavicollis). Antibodies to N. caninum were detected by commercially available competitive-inhibition enzyme-linked immunosorbent assay in one A. flavicolis (0.4%). Three of 427 (0.7%) liver samples were positive for T. gondii by PCR while negative for N. caninum. All embryo samples (n=102) were negative for both T. gondii and N. caninum. The three liver samples positive for T. gondii DNA (two from A. flavicollis and one from A. sylvaticus) were genotyped by 15 microsatellite markers and characterized as type II. To our knowledge, this is the first information about genetic characterization of T. gondii isolates in small mammals from Europe and the first detection of N. caninum antibodies in wild rodents from the Czech Republic.

PubMed | State Veterinary Institute Prague and National Institute of Public Health
Type: | Journal: Scientific reports | Year: 2017

Ongoing evolution of viral pathogens is a significant issue in diagnostic virology employing TaqMan qPCR/RT-qPCR. Specific concerns are related to false negativity due to probe binding failure. One option for compensating for such deficiency is to integrate a second identically labelled probe in the assay. However, how this alteration influences the reaction parameters has not been comprehensively demonstrated. In the present study, we evaluate a TaqMan protocol using two identically labelled hydrolysis probes (simple, LNA (locked-nucleic-acid)) and MGB (minor-groove-binder) modified probes and combinations thereof in a single assay. Our results based on a synthetic amplicon suggest that the second probe does not compromise the TaqMan qPCR/RT-qPCR parameters, which repeatedly and reproducibly remained comparable to those of the corresponding single-probe assays, irrespective of the relative probe orientation, whether opposite or tandem, and probe modifications or combinations thereof. On the other hand, the second probe additively contributed to the overall fluorescence signal. The utility of the dual-probe approach was demonstrated on practical examples by using field specimens. We hope that the present study might serve as a theoretical basis for the development or improvement of TaqMan qPCR/RT-qPCR assays for the detection of highly variable nucleic acid templates.

Bartova E.,University of Veterinary And Pharmaceutical Sciences Brno | Sedlak K.,State Veterinary Institute Prague
Parasitology | Year: 2011

In the Czech Republic, sera from 551 clinically healthy adult slaughtered pigs (females, 6-8 months old) were collected during the first half of June in 2010. Sera were tested for Toxoplasma gondii-specific IgG antibodies by an enzyme-linked immunosorbent assay; samples with more than 50% S/P were considered as positive. The same samples were also analysed for Neospora caninum antibodies using a commercial competitive-inhibition enzyme-linked immunosorbent assay; samples with more than 30% inhibition were considered as positive. Antibodies against T. gondii were found in 198 pigs (36%) in all districts with prevalences ranging from 18% to 75%. Antibodies against N. caninum were found in 16 pigs (3%); positive animals were found in 4 districts with prevalences ranging from 1% to 20%. Indication of mixed infections (concurrent presence of both N. caninum and T. gondii antibodies) was found in 8 (1A.5%) pigs. The results of our study indicate that pigs in the Czech Republic have a relatively high seroprevalence for T. gondii, while they have only a low seroprevalence for N. caninum. Therefore, natural infection with T. gondii seems to be very common in Czech pigs. It is the first evidence of N. caninum antibodies in pigs in the Czech Republic. These results complete data about N. caninum infection in pigs in Europe. © Copyright Cambridge University Press 2011.

Nagy A.,State Veterinary Institute Prague | Cernikova L.,State Veterinary Institute Prague | Krivda V.,State Veterinary Institute Prague | Hornickova J.,State Veterinary Institute Prague
Virus Research | Year: 2012

The objective of our study was to provide a genotype analysis of H7N7 and H7N9 influenza A viruses (IAV) and infer their relationships to co-circulating non-H7 IAV genomes. The H7N7 strains were collected in central Europe (Hungary-1, Czech Republic-1, Slovenia-1 and Poland-4) and the H7N9 in the Czech Republic and Spain between 2007 and 2011. Hand in hand with this effort, a novel IAV genotype visualization approach called digital genotyping was developed. This approach relies on phylogenetic data summarization and transformation into a pixel array called a segment identity matrix. The digital genotyping revealed a complicated genetic interplay between the H7 and co-circulating non-H7 IAV genotypes. At the H7 IAV level the most obvious relationships were observed between one Polish H7N7/446/09 and Czech H7N7/11 viruses which, despite the special and temporal distance of 800. km and 15. months, retained at least 6/8 genome segments. Close relationships were also observed between the Czech H7N9, Polish and Slovenian H7N7 on one hand and Hungarian and Slovenian H7N7 isolates on the other. In addition the former genomes exhibited close interplays with the Czech H6N2/09 and H11N9/10-like viruses. The Czech and Spanish H7N9 genomes were completely different and 6/8 of the Czech H7N9-like segments were traced to either the Czech H3N8/07, H11N9/09 and Polish H7N7/09-like viruses. The results of digital genotyping correlated with the previous observations obtained on the Polish H7N7 isolates. As was demonstrated, the digital genotyping provides a well-arranged and easily interpretable output and may serve as an alternative genotyping tool useful for handling and analysing even a large panel of IAV genomes. © 2012 Elsevier B.V.

Vaclavik L.,Institute of Chemical Technology Prague | Rosmus J.,State Veterinary Institute Prague | Popping B.,Eurofins | Hajslova J.,Institute of Chemical Technology Prague
Journal of Chromatography A | Year: 2010

The use of fast semi-automated method employing direct analysis in real time (DART) ion source coupled to time-of-flight mass spectrometry (TOFMS) for analysis of melamine (MEL) and cyanuric acid (CYA) in milk powder and milk based products has been demonstrated in this study. Simple sample extraction procedure employing methanol-5% aqueous formic acid mixture, which enabled disruption of melamine-cyanurate complex, was followed by direct, high-throughput (30s per run) examination of sample extracts spread on a glass rod by mass spectrometry under ambient conditions, without any prior chromatographic separation. After optimization of instrument parameter settings, limits of detection (LODs) 170 and 450μgkg-1 were achieved for MEL and CYA, respectively. In the final phase of study, the possibility of minimizing spectral interference, thus improving method performance characteristics through the use of ultrahigh resolving power offered by Orbitrap based mass analyzer is demonstrated. © 2010 Elsevier B.V.

Bartova E.,University of Veterinary And Pharmaceutical Sciences Brno | Sedlak K.,State Veterinary Institute Prague | Syrova M.,University of Veterinary And Pharmaceutical Sciences Brno | Literak I.,University of Veterinary And Pharmaceutical Sciences Brno
Parasitology Research | Year: 2010

During January 2007, blood samples were collected from 552 healthy horses from nine different regions of the Czech Republic. Sera were tested for serum antibodies to Neospora caninum by a competitive-inhibition enzyme-linked immunosorbent assay and confirmed by an indirect fluorescent antibody test. The same samples were tested for serum antibodies against Toxoplasma gondii by a latex agglutination test. In total, 131 of 552 (24%) horses reacted positively for Neospora antibodies in competitive-inhibition enzyme-linked immunosorbent assay; seven of them had ≥50% of inhibition. Samples were confirmed in indirect fluorescence test, and only two samples were positive with final titres 50 and 100, while others were negative. Antibodies against T. gondii were found in 125 (23%) horses. This is the first serologic survey for Neospora spp. antibodies performed on horses in the Czech Republic. © 2010 Springer-Verlag.

Maca O.,State Veterinary Institute Prague | Maca O.,Czech University of Life Sciences | Pavlasek I.,State Veterinary Institute Prague
Veterinary Parasitology | Year: 2015

This paper represents the first report of spontaneous infection with Cryptosporidium baileyi and Cryptosporidium meleagridis in the red-legged partridge (Alectoris rufa), as well as the percentage of positive samples and age-associated dynamics of cryptosporidial infections in an aviary in the Czech Republic. The entire infection process was monitored over two semesters (July-December 2012 and 2013) until release of birds for hunting purposes. Coprological examination of 663 pooled fecal samples and 89 post-mortem examinations of red-legged partridges were carried out. Our results indicated that infections with C. baileyi only occurred in 5-7 week-old birds during 2013 (percentage of positivity, 1%) and those with C. meleagridis in 18-22 week (17%) and 17-21 week-old birds (24%) during 2012 and 2013, respectively. Molecular characterization of isolates of C. baileyi and C. meleagridis heat shock protein 70 and actin genes were analyzed in order to support our coprological results. DNA sequence analysis of the 60. kDa glycoprotein gene was used to subtype C. meleagridis. Our findings extend the host range for C. baileyi. © 2015 Elsevier B.V.

Maca O.,State Veterinary Institute Prague | Maca O.,Czech University of Life Sciences | Pavlasek I.,State Veterinary Institute Prague
Parasitology Research | Year: 2016

From July to November 2012, preliminary coprological examinations were carried out on 85 pooled faecal samples of different aged ring-necked pheasants (Phasianus colchicus) (hatches from May until July) from an intensive artificial breeding programme in the Czech Republic. Cryptosporidium oocysts were detected in 12 samples (14.1 %) of ages >12 weeks (August–September). These results were supported by findings of Cryptosporidium baileyi and Cryptosporidium meleagridis oocysts in intestinal, or cloacal contents, and/or the bursa of Fabricius in 9 from 36 examined dead pheasants (prevalence 25 %). We describe in detail the various age groups of pheasants after hatching and present graphically the overall results of coprological examinations, showing pathways of infection of C. baileyi and C. meleagridis during the full rearing seasons of 2013 and 2014. We found very similar mean proportions of Cryptosporidium-positive samples over the entire 2013 period in pheasantry (173 pooled samples tested, 25 positive, 14.5 %) and 2014 (238 samples tested, 43 positive, 18.1 %). All tests were verified as being Cryptosporidium positive in 9 from 219 (prevalence 4.1 %) and 4 from 168 (prevalence 2.4 %) post-mortem examinations. Significantly, C. baileyi was found more frequently in faeces, with positivities ranging from 11.1 to 100 % (4–>16-week-old pheasants). Oocysts of C. meleagridis were detected at ages 6–>15 weeks ranging from 7.1 to 100 % in faeces during the rearing seasons. The burdens of C. baileyi (7 of 14 and 10 of 16) and C. meleagridis (5 of 14 and 7 of 16) for each year, in monitored brooder houses, flight pens and spread across all open areas were recorded. Oocysts of C. baileyi and C. meleagridis obtained from this study, and Cryptosporidium galli (obtained in another aviary from 36-week-old pheasants), were sequenced, and we characterized the highly variable 60-kDa glycoprotein gene of C. meleagridis. These results highlight the real risk of transmission of Cryptosporidium to susceptible wild birds and other potential hosts after termination of rearing and release. © 2016 Springer-Verlag Berlin Heidelberg

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