Isolation of genomic DNA is one of the basic steps in many different molecular analyses. There are a few reports on methods of DNA isolation from milk, but many of them are time consuming and expensive, and require relatively large volumes of raw milk. In this study a rapid, sensitive, and efficient method of DNA extraction from milk somatic cells of various mammals (cattle, sheep, goats, horses) is presented. It was found that milk is a good source of genomic DNA, and to obtain a sufficient amount and quality of DNA, suitable for molecular analysis such as PCR, 10 mL of raw milk is sufficient. Thanks to this method, stress in animals can be reduced during collection of researched material. Therefore, this method could be widely used in molecular analyses.
The aim of this study was to identify the association between single nucleotide polymorphisms (SNPs) in the bovine chemokine receptor (CXCR1) gene and the resistance or susceptibility of cows to mastitis. The analysis of the CXCR1 polymorphism was carried out using polymerase chain reaction restriction fragment length polymorphism analysis for six SNP mutations (c.+291C>T, c.+365T>C, c.+816C>A, c.+819G>A, +1093C>T, and +1373C>A), of which four were located within the coding region and two in the 3'UTR region of the CXCR1 gene. Genetic material from 146 Polish Holstein-Friesian cows was analyzed after dividing into two groups depending on the incidence of clinical mastitis. Identified polymorphisms were in linkage disequilibrium and formed two linkage groups. Three haplotypes (CCCATA, TTAGCC, CTCGCC), forming six haplotype combinations, were detected. The logistic regression showed a significant association between the CC genotype at c.+365T>C and susceptibility of cows to clinical mastitis (P = 0.047). The frequency of haplotype combination 1/1 (CCCATA/CCCATA) was not significantly higher in cows susceptible to mastitis (P = 0.062). Of the identified SNP mutations, only c.+365T>C is a nonsynonymous mutation that induces a change in the coded protein [GCC (Ala) to GTC (Val) at the 122nd amino acid]. This amino acid change can result in changes in receptor function, which may be a reason for the increased mastitis incidence observed in cows with polymorphism at this site.
Aim. This study investigates the changes in hemorheologic parameters resulting from exercise followed by a standard meal.Methods. In twelve moderately active men a period of exercise on a bicycle ergometer for 30 min at 60%was followed by a test meal or by 30 min rest. Venous blood was sampled for further analysis at baseline, after exercise, and after the meal/rest period.Results. The elongation index (EI) was reduced and a marked rise in plasma viscosity was observed after exercise. A significant decrease in half time of total aggregation (T1/2) and a rise in aggregation index (AI) after exercise were observed; however, after the postexercise period these changes were reversed.Conclusion. The present study demonstrates that physical exercise causes several changes in blood rheology parameters, such as an increase of blood viscosity, a decrease in EI and an increase in AI, and a fall in theT1/2values. The meal eaten in the postexercise period caused a further reduction in EI values indicating higher red cell rigidity, but not in plasma viscosity or aggregations indices. Such alterations in hemorheologic parameters should not impair the function of the cardiovascular system in fit and healthy people but it could constitute a serious risk under various pathophysiological conditions.