The focus of this study was microsatellite diversity of crossbred horses raised in Bosnia and Herzegovina. Genomic DNA was extracted from blood samples of 20 individuals (KBA group – 7 individuals, crosses between Bosnian and Herzegovinian mountain horse and Arabian horse; KBR group – 9 individuals, crosses between Bosnian and Herzegovinian mountain and Belgian horses, crosses between Bosnian and Herzegovinian mountain horses and Holstein, crosses between Bosnian and Herzegovinian mountain and Lipizzaner horses and KBN group – 4 individuals, crosses between Bosnian and Herzegovinian mountain horse with an unknown origin of the other parent). The samples were profiled using 17 microsatellite markers. This method consisted of multiplex PCR procedure and generated reasonable amplification across all the loci. All samples were genotyped successfully. Considering all the observed parameters, VHL20 locus showed the highest microsatellite diversity. Locus HMS7 was the least variable in KBR group, while HMS1 locus was the least diverse in KBN group. The highest microsatellite diversity in KBA group was found at AHT5 locus while HTG6 locus was the least diverse. Obtained results suggest that the investigated populations of crossbred horses from Bosnia and Herzegovina are not affected by substantial loss of genetic diversity, as indicated by the presence of reasonably high level of genetic variation. An increase in the inbreeding coefficient and sufficient heterogenity in KBN group indicate occurrence of consanguineous mating. The present research contributes to the knowledge of current status of genetic structure of the investigated crossbred horses.

The focus of this study was on genetic diversity of TB horse population raised in B&H. Genomic DNA was genotyped by using 17 microsatellite markers. A total of 103 alleles were detected. The average number of alleles per locus was 6.059 and effective number of alleles was 3.293. Means of observed and expected heterozygosity were calculated 0.645 and 0.696, respectively. The average PIC values was 0.649 and inbreeding coefficient was 0.090. Based on all observed parameters, ASB2 locus showed the highest genetic diversity while locus HMS2 was the least diverse. These results suggest that the population of TB horses from B&H is not affected by substantial loss of genetic diversity, indicating the presence of reasonably high level of genetic variability.

The serum concentrations of hormones are widely used to diagnose thyroid and adrenocortical diseases in animals. The objectives of the present study were, for the first time, to determine serum thyroxine (T4) and cortisol values in Bosnian and Herzegovinian mountain horses, and to investigate the influences of age and gender on T4 and cortisol levels. Blood samples were collected on the stud farm Borike from 23 apparently healthy horses of both sexes aged between two and twenty-three years. Horses were grouped in three age classes: young mature horses, middle-aged horses and old horses. Serum samples were analyzed for T4 and cortisol concentrations. The T4 values ranged from 6 to 45 nmol/L with a mean ±SD of 22.83±8.32 nmol/L. The cortisol values ranged from 149 to 190 nmol/L with a mean ±SD of 171.57±11.46 nmol/L. Values of T4 decreased with increasing age (p=0.021). Mean value of T4 in young mature horses was 27.60 nm/L, in middle-aged horses was 19.40 nm/L and in old horses was 18.33 nm/L. Values of T4 were significantly higher (p=0.031) in females (24.88 nm/L) than in males (18.14 nm/L). Negative significant correlation between age and hormone concentration was observed for T4 (r = -0.431; p = 0.040). Differences between gender or age classes for cortisol values were not significant. The mean total serum T4 and cortisol values in Bosnian and Herzegovinian mountain horses were in the normal reference ranges for horses, and both age and gender significantly affected T4 levels.

Herzegovinian donkey is a very important animal resource of Bosnia and Herzegovina. There have been no works attempted at determining the normal values of hematological parameters of Herzegovinian donkey. For this reason, the objective of the present study was to investigate some hematological parameters in Herzegovinian donkey. The research was performed on 30 apparently healthy donkeys (18 female and 12 male) of ages from 1 to 20 years. Blood samples (3 ml) were obtained by jugular vein puncture (vena jugularis externa) in vacuum tubes with EDTA. The mean value of hematocrit was 29.19 %, hemoglobin concentration 10.6 g/dl, mean corpuscular hemoglobin concentration 36.33 g/dl, white blood cells 9.33 x109/L, granulocytes (109/L) 5.45 x109/L, granulocytes (%) 59.47%, lymphocyte/monocyte (109/L) 3.89 x109/L, lymphocyte/monocyte (%) 40.53% and platelet cells 148.97 x109/L. Parameters were determined using an automated analyzer IDEXX QBC VET AutoRead. Data were analyzed by SPSS V 15. All hematological parameters (except platelet cells) were consistent with the recommended reference ranges for donkeys, and the values found in literature so far. Platelet cells values were much lower than in the literature for the other donkey breeds and the recommended reference ranges for donkeys. The slight differences found between our results and those reported in the previous works confirm the need for further studies to investigate the reference values of hematological parameters of Herzegovinian donkey. This work is a contribution to the study of hematological parameters of Herzegovinian donkey, and we expect these data to be applied to the further studies.

21 RRJVS| Volume 2 | Issue 1 | March, 2016 Genetic Diversity of Arabian Horse from Stud "Borike" (Bosnia and Herzegovina) Using Microsatellite Markers Dunja Rukavina1*, Danica Hasanbašić1, Adaleta Durmić-Pašić2, Belma Kalamujić2, Amir Zahirović3, Jasmin Ramić2, Naris Pojskić2 1Department for Biology, Veterinary Faculty, University of Sarajevo, Zmaja od Bosne 90. Bosnia and Herzegovia. 2Institute for Genetic Engineering and Biotechnology, University of Sarajevo, Zmaja od Bosne 8. Bosnia and Herzegovia. 3Department for Internal Diseases, Veterinary Faculty, University of Sarajevo, Zmaja od Bosne 90, Bosnia and Herzegovia.

In the present study, for the first time we investigated the genetic diversity among horse breeds from Bosnia and Herzegovina: potential Bosnian and Herzegovinian mountain horse, Arabian horse, Thoroughbred horse and crossbreeds, with special emphasis on the gene pool of potential Bosnian and Herzegovinian mountain horse. In total, 138 animals were genotyped for 17 microsatellite loci. Compared to the other breeds, potential Bosnian and Herzegovinian mountain horse showed quite a high genetic variability. The mean number of alleles was 14.1765. The average observed heterozygosity was 0.6589 and the expected heterozygosity was 0.8451. The mean value of polymorphic information content was 0.8286. The results of AMOVA test showed 8,44% of genetic variation among populations. The highest genetic variation within population was showed by potential Bosnian and Herzegovinian mountain horse (27.13). The same breed showed the highest individual variation (17.35). Overall FST value showed high level of the genetic differentiation among breeds (8.87 %), and the pairwise FST values were all significant. Highest inter-group genetic differentiation was observed among Arabian horse and Thoroughbred horse (groups of pure breeds) and potential Bosnian and Herzegovinian mountain horse. The results show that the potential Bosnian and Herzegovinian mountain horse has a high in-breed variability, more than could be expected. In the gene pool of potential Bosnian and Herzegovinian mountain horse, a part of the gene pool of other breeds is present. Also, these results show that there are very good preconditions for the revitalization of the gene pool of potential Bosnian and Herzegovinian mountain horse. Keywords: Horse, genetic diversity, microsatellites

Ionising radiation is known to cause chromosomal instability, which is observed as increased frequency of chromosomal aberration and micronuclei. These are listed as reliable criteria in biological dosimetry. Numerous experiments conducted on both animal and plant models demonstrated that increase in radiation dosage is followed by increased mutation frequency, and that mutations occur even at the lowest exposure. We used horse blood in vitro irradiated by low doses of ionizing radiation. Cultivation of peripheral blood lymphocytes and micronucleus test were used as biomarkers of genetic damage. The observed aberrations were recorded and classified in accordance with the International System of Cytogenetic Nomenclature. Micronuclei were identified on the basis of criteria proposed by Fenech et al . (8). Analysis of chromosomal aberration showed increased frequency of aberrations in blood cultures exposed to 0,1 Gy and 0,2 Gy compared to the controls. Microscopic analysis of chromosomal damage in in vitro micronucleus test revealed that the applied radiation dose induced micronuclei while no binucleated cells with micronuclei were found in lymphocytes that were not irradiated. In this paper we analysed the influence of low dose ionising radiation on frequency of chromosomal aberration and micronuclei in horse lymphocytes following in vitro exposure to X-rays (0,1 Gy and 0,2 Gy). Key words : chromosomal aberrations, micronuclei, ionising radiation, horse lymphocytes

In this study we investigated the influence of low dosage X - ray irradiation on the incidence of chromosomal damage and changes in mitotic index (MI) in cultured peripheral lymphocytes of the Bosnian and Herzegovinian mountain horse following in vitro irradiation. X-ray irradiation induced a dose-dependent decrease in MI but only the dose of 0.5 Gy induced a significant decrease (p<0.05) in comparison with the control and other dose groups. The analysis of chromosomal damage revealed a clear dose-dependent increase in the incidence of chromosomal damage per metaphase. Significant differences (p<0.05) were detected by analysis of variance and the LSD test confirmed significant differences between cells that received 0.2 Gy and 0.5 Gy when compared to the control cells and cells that received 0.1 Gy. However, Scheeffes' test assigned significance only to the differences established between the cells that received 0.5 Gy and the other groups of lymphocytes. .

Knowledge of spontaneous aberrations, namely, of their frequency in non-irradiated cells is of paramount importance not only in cytogenetic research, but also in contemporary animal production. The paper deals with research on spontaneously emerged chromosomal aberrations in the peripheral blood lymphocytes in the cattle of 'Busa' breed. To obtain metaphase chromosomes the conventional method of lymphocyte cultivation was used, albeit slightly modified and adapted to the examined animals and the laboratory conditions. The research findings indicate that a certain percent of spontaneously emerged chromosomal aberrations of chromatid type (gap and break) have been found in the peripheral blood lymphocytes in the cattle of 'Busa' breed.

Within a comprehensive study conducted on the Bosnian- Herzegovinia mountain horse breed, a research on the relative frequency of micronuclei (MN) after in vitro horse blood irradiation was carried out. Experiments on cytogenetic dosimetry were also conducted on pigs. The results of MN test were presented in terms of MN in binucleate (BN) cells. In control samples the percent of MN is very low in relation to irradiated samples where a higher number of MN cells was asserted, as well as a higher number of MN in BN cells. We could also observe that a number of BN cells with MN is proportionally increased with the radiation dosage. The MN test could provide additional data on the level of radiation damage (depletion) in the lymphocytes of the peripheral blood in domestic animals.