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Small ruminant lentiviruses - analysis of disease prevalence and distribution of SRLV genotypes in sheep and goat farms in the Czech Republic
VERNEROVÁ, Kateřina
Maedi-visna in sheep, and arthritis and encephalitis in goats, are globally widespread and progressive inflammatory diseases caused by retroviruses belonging to the small ruminant lentivirus (SRLV) group. They cause lifelong infections in affected animals, characterised by slow progression to overt disease and are always fatal. The aim of this study was to determine the prevalence of SRLV disease in the Czech Republic, to map the genotypic representation of SRLV using phylogenetic analysis, and to analyse TMEM154 as a selected candidate marker of SRLV resistance in sheep and goats. A total of 3 410 sheep and goat blood samples were collected from 21 flocks. The seroprevalence of maedi-visna in sheep was found to be 19.9% (556/2801), and the seroprevalence of arthritis and encephalitis in goats was 14.1% (86/609). All seropositive animals were tested by nested polymerase chain reaction (nPCR) for the presence of proviral DNA. Phylogenetic analysis identified the SRLV genotype in 77 sequences, of which 60 sheep and goat samples were genotype A and 17 sheep samples were genotype B. While all genotype B sequences were classified as subtype B2, the group of genotype A isolates showed higher variability and were related to subtypes A2 and A3. In addition, 40 seropositive and 50 seronegative sheep and goat samples were randomly selected to design a methodology for LAMP (loop-mediated isothermal amplification) diagnosis of SRLV in sheep and goats. Seronegativity was clearly confirmed by the LAMP method in all samples, and seropositivity was confirmed in 31 out of 40 cases in both sheep and goats. 605 sheep and 60 goat samples were randomly selected for TMEM154 genotyping. Most seropositive animals were heterozygous EK (61%), 58% were homozygous EE, and 45% were homozygous KK. In sheep, all 3 genotypes were identified, while all goats were homozygous EE.
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Genetic polymorphism of selected loci in relationship to technological traits of meat
VERNEROVÁ, Kateřina
Within the framework of this diploma, a genotypization of 162 beef cattle of the ČESTR commercial breed from was performed in order to analyze the potential influence of gene coding calpain I (CAPNI) on the resulting beef tenderness. CAPN1 present on the BTA29 chromosome, was selected in previous studies as a candidate gen for QTL. Polymorphism of CAPNI gene was studied using PCR/RFLP method and CAPN530 marker. Genotype identification resulted from this procedure. A 341 bp long fragment was present in homozygotes AA, two fragments of 195 and 146 bp were present in homozygotes GG, and three fragments of 341, 195 and 146 bp were detected in heterozygotes AG. The genotypization output was subsequently statistically evaluated. 11 homozygotes AA, 62 homozygotes GG and 89 heterozygotes AG were detected in the analyzed beef cattle population. More frequent G allele occurred in the set with the frequency of 0,657 and A allele with the frequency of 0,343. Warner-Bratzler shear test was employed to determine beef tenderness based on the shear force attribute. Samples of raw and grilled beef aging 1, 14 and 28 days after the slaughter were analyzed. Statistical methods were used to evaluate relationship between genotype and detected amount of shear force. In case of raw beef, only the samples aging 1 day after the slaughter showed significant difference in the shear force. The most positive results for the given parameter were achieved in case of AG genotype. Samples aging 14 and 28 days after the slaughter showed no difference in shear force and related beef tenderness, indicating no genotype influence. In case of grilled beef, no significant difference in sheer force indicating possible genotype influence was detected at any day of analysis. Genotype influence on grilled beef tenderness within the test animal population is minimal and statistically inconclusive.
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The occurrence of Fascioloides magna in deers in South Bohemia
VERNEROVÁ, Kateřina
Giant liver fluke Fascioloides magna is the biggest represenative of flukes in the Czech Republic. This originally North American parasite was brought to Europe with wapiti deers in 1960s. Since that time Fascioloides magna has extended the area of its appearance and its host spectrum a lot. The first appearance of this parasite in the Czech Republic was noted for the first time at a fallow deer which was hunted down in Middle Bohemia in 1930. However in the early 1960s the number of positive findings started to increase at hoofed. In the last decade was observed the expansion of areas of appearance of this fluke in South Bohemia. That is why I decided to make examinations to evaluate the recent appearance of F. magna at deer family in the South Bohemia from March 2009 to January 2010. 74 samples of dropping of a deer from three different areas (Českobudějovicko, Třeboňsko and Šumava national park) were collected and examined and only in the sample from Šumava national park were proved the ovulums of F. magna. Then I examined 11 livers of deers which were hunted down in Šumava national park. In all these livers were found macroscopic changes. In all these examined livers was also finding of fluke F. magna in number from 1 to 72. From the reached results follows that the problems of appearance of F. magna is in the Czech Republic still actual and it is necessery to count with the extending area of appearance of this parasite in the future. Edification among hunting community and the use of information from trained people who make the primary examination of animals after hunting down are possible ways to catch up appearance and expansion of this serious parasite at hoofed.
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