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Genotypization of selected candidate markers for resistance against lentiviral infections of small ruminants
FARKOVÁ, Barbora
Small Ruminant Lentiviruses (SRLV) known as maedi-visna in sheep, and viral arthritis and encephalitis in goats are widespread worldwide and cause significant economic losses. Lentiviral diseases have been included in the current Animal Health Control and Vaccination Guidelines for Breeding Controls. Control programs are generally based on serological screening and subsequently serologically positive animals are removed from breeding. In the literature review I write about important lentiviruses, including those that have not been analyzed. The aim of this work was to screen for the occurrence of lentiviral infections in small ruminants in the Czech Republic and the genotyping of selected candidate markers and finding the genotype related to the possibility of gene resistance. In total, over 3200 blood samples were collected from sheep and goats from breeders all over the Czech Republic. Genotyping was carried out in herds where serologically positive animals were detected. Amplification of the DNA sequence of interest was performed by PCR (Polymerase Chain Reaction) analysis, followed by genotyping by sequence analysis, and the results were statistically evaluated. In my work, I came to the conclusion that in the Czech Republic the occurrence of 2 out of 5 possible SRLV genotypes (genotype A and B) and distribution of different subtypes in sheep and goats has been detected so far. The greatest differences were in the sequence of genotype A, where one group of isolates showed the possibility of classification into a completely new subtype. In sheep, all three genotypes of the TMEM154 gene were identified. The incidence of serologically positive was statistically significantly higher in animals with a risk genotype. Because of the possible mutation of the virus and the emergence of a new subtype of genotype A, the virus is more likely to adapt to attack and infect individuals with a more resistant genotype. Only one genotype has been demonstrated in goats, so most likely there is no marker associated with resistance to SRLV infections.
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PCR-based detection of hidden carriers of cataracts in dogs
FARKOVÁ, Barbora
The hereditary cataract is one of the most common eye disease in dogs. The expansion of this disease in the Staffordshire bullterrier breed has been so massive that in the Czech Republic was introduced the rule of mandatory testing of at least one of a breeding pair. This is a degenerative disease of the lens causing total blindness of the affected animal within three years. Since some time ago there are no more dogs affected by the disease in the Czech Republic, there are however still hidden carriers which need to be discovered to the complete extinction of the disease in the genome. The goal of this study was to test simple ways of collecting biological samples, try them in practice and to verify whether they are suitable for the DNA isolation and also to test an alternative method of molecular detection of this disease. In total there have been 23 buccal swabs collected from male and female Staffordshire Bullterrier examples. The detection of the hidden carriers of the hereditary cataract was carried out by PCR analysis with specific primers. The obtained amplicons were detected by both gel and chip electrophoresis and by using fragment analysis. This detection of the carriers was based on the presence of two amplicons (heterozygotes). I came to conclusion that to detect hidden carriers it is neccessary to use the fragment analysis because of the difference of only one base in the reference section of DNA. Neither gel nor chip electrophoresis does provide sufficiently high resolution and it is not possible to detect two fragments that differ only by one bp. As the most appropriate sampling method I have chosen the buccal smear by cytological brush followed by isolating the DNA by Chelex with purification of the sample subsequently.
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Pathogenic Microorganisms in Meat Products
FARKOVÁ, Barbora
The aim of this work is the analysis and description of microorganisms occurring in meat products. The work is by definition enter the search character, so the method chosen as the research literature analysis and the subsequent description of findings. The first chapter focuses on the characteristics of microorganisms and their distribution in several respects. Chapter 2 is already covered by specific genera of microorganisms that have been using a wide range of literary sources characterized in detail, including representatives who may be present in meat and meat products. Introduction of Chapter 2 is devoted to foodborne illness ? intoxication and toxic infection, which can occur in humans after ingestion of improperly cooked food or due to poor storage. One of the subsections of Chapter 2 is focused on fungi, which are in search of microbiological work, often neglected, although they are very important pathogens and undesirable elements in food. Another chapter is devoted to meat technology and includes terminology that is used in this field. In addition, summarizes the basic requirements, preconditions and standards associated with the meat industry.
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