The bovine kappa-casein (κ-CN) is a phospho-protein with 169 amino acids encoded by the CSN3 gene. The two most common gene variants in the HF breed are CSN3*A and CSN3*B while CSN3*E has been found with lower frequency. The aim of this study was to optimize a laboratory method for genotyping of these three alleles as well as to determine their genotype and allele frequencies in the HF cattle population in the Republic of North Macedonia. Genomic DNA was extracted from full blood from 250 cows. The target DNA sequence was amplified with newly designed pair of primers and the products were subjected to enzymatic restriction with HindIII and HaeIII endonucleases. Genotype determination was achieved in all animals. The primers successfully amplified a fragment of 458 bp and the digestion of this fragment with both endonucleases enabled differentiation of five different genotypes with the following observed frequencies: AA (0.39), AB (0.29), BB (0.16), AE (0.10), and BE (0.06). The estimated allele frequencies were: CSN3*A (0.584), CSN3*B (0.336) and CSN3*E (0.08). The observed genotype frequencies differed significantly (P<0.01) from those that would be expected under HW equilibrium, while the fixation index (F=0.17) indicated moderate heterozygosity deficiency. Nevertheless, the CSN3*B allele was present with relatively high frequency which should be used to positively select for its carriers, since increasing its frequency could help to improve the rheological properties of the milk intended for cheese production.
Motility patterns of spermatozoa are an indicator of their fertilizing capacity. Reduced glutathione (GSH) has been reported to induce positive effects on the biological quality of the frozen-thawed spermatozoa. The aim of this study was to investigate the effects of GSH addition to semen extender based on the following kinetic parameters: continuous line velocity (VSL), average path velocity (VAP), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), linearity (LIN), straightness (STR), beat cross frequency (BCF), total motility (tMOT) and progressive motility (pMOT), and to appoint them as indicators of its presence. A soybean-based extender was used for dilution of the semen samples, fractioned in two parts, one containing GSH (5 mMol/ml) and second without GSH. The ejaculates (n=48) were collected from two rams (January - May, 2013) which were classified in two groups according to the used extender: Group 1 (with GSH, n=24) and Group 2 (without GSH, n=24), and then frozen in liquid nitrogen on -196°C degrees in a programmable freezer. Assessment of the samples has been performed post-thawing (30 sec. at 37°C) on CASA equipment, acquiring kinetic parameters. Results showed that only VSL and BCF have a statistically significant difference between group1 and group 2 (102.98±15.13 vs. 88.47±20.63, t=2.77, p<0.01 and 32.01±2.68 vs. 89.47±2.92, t=3.13, p<0.01, respectively). The summary of the investigation concludes that none of the kinetic parameters could be appointed as indicators that confer the positive effect of GSH as an additive to soy-bean semen extender.
The illegal poisoning of dogs and other domestic and wild animals presents a worldwide problem causing animal suffering and R. Macedonia is not an exeption. The goal of this study is to make a comparison of the results from the histopathological examination conducted among poisoned dogs in the Republic of Macedonia. Morphological and histopathological changes in poisoned dogs were investigated for a period of 10 years. The examination was performed on 31 dogs, 13 of which were home kept, 7 were street dogs and 11 of unknown origin. The most significant necropsy findings concerned the inflammatory and necrotic processes of the gastrointestinal tract. The histopathological changes were mainly located in the kidneys, stomach, intestines and the lungs.
Artificial insemination (AI) as a part of assisted reproductive technologies represents the oldest and most widespread method used to accelerate genetic progress in all domestic animals. After its first implementation in ovine reproduction and almost 80 years afterward, AI is continuously used for improving the genetic merit, utilizing either fresh or short-time chilled semen. Nevertheless, regardless of the semen used for insemination, the conception rate (CR) is still lower in comparison to natural service. At least two factors are commonly thought to limit the success of the AI and reduce the CR: (1) failure of placing the semen directly into the uterus due to the specific anatomic structure of the ewe’s cervix; (2) lower viability of ram spermatozoa during cryopreservation (<30% progressively motile spermatozoa after thawing). This review elaborates on recent studies that aimed to achieve acceptable CR through the implementation of cervical or intrauterine insemination: deep intracervical, intrauterine trans-cervical, and intracornual. Several hormonal treatments (oxytocin, estrogen, or prostaglandin) were evaluated on inducing cervical dilation that facilitates insemination. A comprehensive analysis was given to the effects of several antioxidants (GSSG, GSH, and cysteine) supplemented in ram semen-freezing media. Sex-sorted ram semen fertility rate results were presented from our studies.
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