KAPPA - CASEIN GENE POLYMORPHISM (CSN3) AND ITS EFFECT ON MILK PRODUCTION TRAITS

The aim of this study was to evaluate the relationship between the polymorphism in the kappa - casein gene and milk production traits in crosses of Simmental and Holstein cattle breeds. A total of 80 hair roots samples were used to investigate how CSN3 gene polymorphisms affect milk production traits. The polymorphism of kappa - casein gene on bovine chromosome 6 was genotyped by using the polymerase chain restriction fragment length polymorphism. The effects of polymorphisms were analysed using SAS 9.1 software. Digestion of PCR products with restriction enzyme HindII revealed two alleles: allele A gave one fragments, 443 bp in length and allele B gave two fragments of 348 and 95 bp. The predominant allele was A with observed frequency 0.76. In population were detected all three genotypes AA, AB and BB.  The most frequent genotype was AA with frequency 0.575. Results from the statistical association analysis between CSN3 genotypes and milk production parameters – milk, protein and fat yield (kg) in standard length of lactation were no significant.Key words: dairy cattle, kappa - casein gene, milk production, polymorphis

SNPs analyses of the bovine LEP and PIT-1 genes by multiplex PCR-RFLP method and their effect on milk performance traits in Slovak Simmental cattle

The aim of this study was detection of polymorphisms in leptin (LEP) and pituitary specific transcription factor (Pit-1) genes and their evaluation as a genetic markers affected milk performance traits in population of Slovak Simmental cows. The analyzed SNP of LEP gene is located in intron 2 on bovine chromosome 4 and SNP of Pit-1 gene in exon 6 on bovine chromosome 1. The total numbers of blood samples were taken from 288 samples of Slovak Simmental cows. Bovine genomic DNA was isolated by phenol-chloroform extraction method and used in order to estimate LEP/Sau3AI and Pit-1/HinfI genotypes by means of MULTIPLEX PCR-RFLP method with using Sau3AI restriction enzyme for LEP and HinfI restriction enzyme for Pit-1. In the population of Slovak Simmental cattle were detected genotype AA (390 and 32 bp), genotype AB (390, 303, 88 and 32 bp), genotype BB (303, 88 and 32 bp) for LEP gene and genotype AA (260 bp), genotype AB (260, 190 and 70 bp), genotype BB (190 and 70 bp) for Pit-1 gene. The predominant alleles were A with observed frequency 0.8385 and B with 0.7743 for LEP and Pit-1, respectively. In both analyzed polymorphisms were frequent animals with dominant homozygous genotype. Frequency of SNP LEP/Sau3AI AA genotype was 0.7014 and SNP Pit-1/HinfI BB genotype was 0.6007. The statistical analysis show significant effect only between LEP/Sau3AI genotype and milk yield in standard length of lactation (P≤0.05), with A as a desirable allele. In genotype comparison produced AA genotype cows significantly more milk, with difference amounting 457.9 and 293.9 kg and therefore this animals might have potentially positive effect on milk yield. The other results from the statistical association analysis between LEP/Sau3AIand Pit-1/HinfI genotypes and milk production parameters – milk, protein and fat yield (kg) in standard length of lactation were not significant

Ascorbic acid supplementation suppresses cadmium-derived alterations in the fission yeast Schizosaccharomyces pombe

Cadmium (Cd) a highly toxic environmental pollutant, that does not have any physiological function in the organism, represents a great concern for human health as it can be easily transported from its environmental sources to the food chain. Food, water, and air are the major sources of Cd exposure to the population. Cd-mediated impairments of the basic cellular properties largely depend on its ability to enhance the formation of reactive oxygen species (ROS) and thus triggers oxidative stress to the cell. With the use of fission yeast Schizosaccharomyces pombe (S. pombe) as the model organism, we have analyzed the impact of Cd on the cell growth intensity, as it represents the fundamental feature of all living organisms. Cells were incubated with different Cd concentrations for 3, 6, and 9 hours to investigate the effect of Cd on cell growth in a time and dose-dependent manner. Further possible Cd-derived alterations, as the peroxidation of membrane lipids or the functional impairment of the enzymatic antioxidant protection mechanisms, were investigated by determination of the MDA content and via catalase (CAT) activity detection. Moreover, ascorbic acid (AsA) pre-treatment was subjected to investigate the assumed positive effect of AsA against Cd toxicity. We show here on one hand that cells suffer under the influence of Cd, but on the other hand, they substantially profit from AsA supplementation. Because S. pombe is known to shares many molecular, and biochemical similarities with higher organisms, the effect of AsA in cadmium toxicity elimination might be expected to a similar extent also in other cell types

Rabbit myostatine gene polymorhism (c.747+34C>T and c.194A>G) as a marker for meat production

Myostatin gene (MSTN or GDF-8) affects both, the amount and composition of muscle fibers. MSTN plays a key role in muscle growth and it has applications in breeding and animal husbandry. Polymorphic variants of this gene are associated with growth rates and carcass yield of different species. This study is focused on two single nucleotide polymorphisms: MSTN SNP c.747+34C>T and MSTN SNP c.194A>G and their effect on selected production traits. Genomic DNA was extracted from whole blood. Polymorphic variants of MSTN gene were detected using PCRRFLP method. Association studies revealed significant positive effect of allele T (c.747+34C>T) and allele G (c.194A>G) on meat performances in tested rabbit population

Leptin receptor and ghrelin genes polymorphisms in relation to the metabolism of lipids

The aim of this work was to analyse genetic polymorphisms in genes encoding leptin receptor (LEPR) and ghrelin (GHR) as genetic markers of metabolic disorders in human nutrition. Genomic DNA was obtained from in total 84 human blood samples. Effect of analysed genetic markers was evaluated for three biochemical parameters: total cholesterol, HDL and LDL cholesterol. The PCR-RFLP method was used for identification of SNPs in LEPR (Gln223Arg) and GHR (171T/C) genes. In analysed population prevalence of heterozygous LEPRAG (47.62%) and GHRCT (40.48%) genotypes was observed. Frequency of LEPRA and LEPRB alleles were 0.55 and 0.45, respectively. Similar the GHRC allele had only slight predominance than GHRT allele (0.54/0.46). In population was found higher level of observed heterozygosity across loci (0.44). For both SNPs was found high effective allele number (1.98) which was also transferred to the median level of polymorphic information content (0.37). Association analysis of LEPR and GHR genotypes effect on selected biochemical parameters was performed using GLM procedure. Significant association was found only for levels of LDL cholesterol (P<0.01). Our study shows that both genes are involved in nutritional status and therefore can be considered as candidate genes of lipids metabolism disorders and obesity

Genetic diversity in populations of Slovak Spotted cattle based on single nucleotide polymorphisms analyses

The aim of this study was to identify SNPs in leptin (LEP), leptin receptor (LEPR) and growth hormone (GH) genes in order to analyze genetic diversity of Slovak Spotted cattle. The total numbers of blood samples were taken from 353 Slovak Spotted cows originating from four farms. Genomic DNA was isolated by phenol-chloroform extraction method and analyzed by PCR-RFLP method. After digestion with restriction, enzymes were detected in whole population of cow's alleles with frequency: LEP/Sau3AI A 0.84 and B 0.16 (±0.0152); LEPR/BseGI C 0.95 and T 0.05 (±0.0089) and GH/AluI L 0.70 and V 0.30 (±0.0188). Based on the observed vs. expected genotypes frequencies populations across loci were in Hardy-Weinberg equilibrium (P\>0.05). Predominant for SNP LEP/Sau3AI was AA genotype (0.70), for SNP LEPR/T945M CC genotype (0.91), and LL genotype (0.48) was most frequent for SNP GH/AluI. The observed heterozygosity of SNPs across populations was also transferred to the low or median polymorphic information content 0.24 (He 0.28), 0.08 (He 0.09) and 0.33 (He 0.47) for LEP, LEPR and GH genes, respectively. Within genetic variability estimating negative values of fixation indexes FIS (-0.09-0.05) and FIT (-0.07-0.03) indicating heterozygote excess were observed. The value of FST indexes (0.018-0.023) shows very low levels of genetic differentiation in allele frequencies of loci among evaluated subpopulations. The low values of genetic distances (0.0018-0.0159) indicated high genetic relatedness among animals in subpopulations caused probably by common ancestry used in breeding program at farms