Bats are a natural host for a number of viruses, many of which are zoonotic and thus present a threat to human health. RNA viruses of the family Filoviridae, many of which cause disease in humans, have been associated with specific bat hosts. Lloviu virus is a Filovirus which has been connected to mass mortality events in Miniopterus schreibersii colonies in Spain and Hungary, and some studies have indicated its immense zoonotic potential. A die-off has been recorded among Miniopterus schreibersii in eastern Bosnia and Herzegovina for the first time, prompting the investigation to determine the causative agent. Bat carcasses were collected and subjected to pathological examination, after which the lung samples with notable histopathological changes, lung samples with no changes and guano were analyzed using metagenomic sequencing and RT-PCR. A partial Lloviu virus genome was sequenced from lung samples with histopathological changes and found to be closely related to Hungarian and Italian virus sequences. Further accumulation of mutations on the GP gene, coding the glycoprotein responsible for cell tropism and host preference, enhances the need for further characterization and monitoring of this virus to prevent spillover events and protect human health.

OBJECTIVE Canine hip dysplasia (CHD) is a global disease and an improved diagnostic approach to this orthopedic condition is needed, with an emphasis on inexpensive and minimally invasive techniques. This research aimed to determine the differentiating potential of selected biochemical markers in serum between healthy dogs and dogs with hip dysplasia in a breed-specific study that involved the Tornjak dog population. ANIMALS 99 Tornjak dogs radiographically categorized (Federation Cynologique Internationale procedure and scoring scheme) between December 2019 and April 2021, as having no sign of hip dysplasia or near normal hip joints (nondysplastic group; n = 51) vs mild, moderate, or severe hip dysplasia (dysplastic group; 48). METHODS Serum concentrations and enzyme activity of the biochemical markers hyaluronic acid (HA), procollagen II C-terminal propeptide (PIICP), and metalloproteinase 9 (MMP9) were compared among adult dogs with or without radiographic hip dysplasia. Statistical tests used to examine the differentiating potential of biochemical markers in Tornjak dog groups were assessed using the Mann-Whitney U test, logistic regression, and receiving operating characteristics (ROC) analysis. RESULTS Tornjak dogs with radiographic CHD had significantly lower serum concentrations of HA and higher concentrations of PIICP and MMP9 activity compared to dogs with radiographically normal hips (P < .05). Selected biochemical markers could distinguish dogs with radiographic CHD from those without CHD with high sensitivity and specificity. CLINICAL RELEVANCE These data suggest that the diagnostic technique of measuring serum concentrations of HA, PIICP, and MMP9 activity has a selective ability to distinguish dogs with dysplastic from dogs with normal hips.

The infection with SARS-CoV-2 virus in cats and dogs raised issue of human-to-animal transmission of SARS-CoV-2 in domestic pets in close contacts with their owners. Our study was designed to research this in the framework of Bosnia and Herzegovina. Using ELISA, AFIAS fluorescent immunoassay, RT-qPCR and WGS on Nanopore MinION platform with ARTIC Network Amplicon sequencing protocol for SARS-CoV-2, we showed that three out of thirteen dogs and one out of five cats from the households with confirmed human cases of COVID-19 in Bosnia-Herzegovina were infected with SARS-CoV-2. The high viral RNA load was detected in samples collected from a 4-year-old male Havanese (Ct = 12.52), a 6-year-old German Shepherd (Ct = 21.36) and a 9-year-old female American Staffordshire terrier (Ct = 25.74). The antibody response in dogs and one cat was observed. The viral genetic sequences from dogs were identical to the sequences detected in the owners suggesting the human-to-animal transmission of the virus. These findings, especially the low initial Ct values detected, from the public health perspective additionally stress the need for precautionary measures to protect both humans and animals.

Background: Thimerosal is an organomercury compound with high mercury content. Thimerosal is located in the market as an effective bacteriostatic in a series of pharmaceutical products, ophthalmic and nasal products, immunoglobulin preparations, and as a preservative in vaccines. Aims and Objectives: Since it is a compound with living content, this study aimed to examine the genotoxic and cytotoxic potential of thimerosal in human lymphocytes culture. Materials and Methods: We used chromosome aberration analysis and cytokinesis-block micronucleus cytome (CBMN-Cyt) assay to test its genotoxic and cytotoxic potential in human lymphocyte culture. Results: Results showed that the frequency of structural chromosome aberrations and CBMN-cyt assay was significantly increased in treated cultures (1 μg/ml and 0.5 μg/ml) compared to the negative control. Conclusion: Obtained results and statistical analysis show that thimerosal is genotoxic and cytotoxic in human lymphocytes in tested concentrations.

The primary purpose of this study was to determine the frequency and types of chromosomal aberration in infertile couples. First-trimester pregnancy loss, as well as infertility, is a prevalent issue among many couples, and the main reason they investigate is their chromosomes in that case. Two hundred and sixty-six couples came to the Center for Genetics of Medical Faculty-University of Sarajevo for chromosomal cytogenetic analysis due to infertility or a more significant number of spontaneous abortions. A karyotype has been made to each couple, using GTG technique from cultures of lymphocytes from peripheral blood. The overall incidence of chromosomal abnormalities was 15 out of 266 couples (9%). They were divided by primary diagnose as couples with recurrent spontaneous abortions 107 and infertility couples 159. The first group has found chromosomal changes in four males and one female, whereas, in the second group, it has found chromosomal changes at seven females and three males. Structural and numerical chromosomal changes did not differ from relevant updated references from other countries and were the leading cause for infertility and recurrent spontaneous abortions. This study illustrates the incidence and distribution of chromosomal abnormalities among couples with recurrent miscarriage and infertility. Genetic testing and counseling are essential for couples with an abnormal karyotype before deciding about further reproductive options.

Infertility is one of the most significant human health problems of the reproductive years. The causes of infertility are diverse and numerous-including non-genetic and genetic factors. A review of this case confirmed this. A balanced translocation was found in two siblings diagnosed with azoospermia. After being unable to conceive, the older brother and his wife had two in vitro fertilization (IVF) failed. At the same time, his younger brother and his wife had one IVF.  After the cytogenetic analysis was performed in both pairs, it was shown that the cause of their infertility was the same balanced translocation in the brothers. The female showed a regular (46, XX) karyotype, whereas the male was found to carry balanced reciprocal translocation [46, XY, t(8;17)(p23;q11)]. This case support that cytogenetic analysis is still the first and basic diagnostic analysis of patients with azoospermia and other reproductive problems.

Reptiles, especially turtles, are becoming increasingly popular as pets. The haematological evaluation of turtles is an irreplaceable diagnostic tool in veterinary practice. However, the morphologic distinctiveness of turtle blood limits the use of electronic cell-counting devices, making time-consuming, manual counting techniques and evaluation of blood smears necessary. Many samples are dispatched to a laboratory over long distances, where a delay of 24 h or more may occur. At weekends, this interval may exceed 48 h. The objective of the present study was to determine the effect of storage duration at refrigerator temperature (4 °C) on the counts of red blood cells (RBC) and white blood cells (WBC), and on the mean corpuscular volume (MCV) and packed cell volume (PCV) in blood samples from healthy adult red-eared sliders. Blood samples were collected by venipuncture from the occipital venous sinus from six apparently healthy adult red-eared sliders, aged 2 to 4 years. Blood samples were analysed immediately after sampling to obtain the baseline value (BV) of the red blood cell count, white blood cell count and packed cell volume percentage. Blood was stored at 4 °C and the haematological analyses were performed after 24h, 48h and 72h. The results showed the same level of stability for RBC and WBC count, and MCV values during 72 hours of storage at 4 °C and for PCV during 48 hours. Handling of blood samples, and duration of storage of the blood samples can significantly influence the results/values of haematological tests. Consequently, the obtained values of the determined haematological parameters of improperly stored or handled blood samples can give a misleading interpretation of the results on the animal’s health status.

In the present study modern technology of DNA extraction and automatic genotyping was applied in Bosnian and Herzegovinian autochthonous horse breed by using 17-Plex horse genotyping kit. The study was aimed at investigating usefulness of the 17-plex STR Kit for Bosnian mountain horse genotyping and establishing highly useful microsatellite markers system for genetic diversity studies in Bosnian mountain horse breed. Genomic DNA was extracted from whole blood collected from 22 unrelated Bosnian mountain horse specimens. A total of 95 alleles were detected. Average number of detected alleles per locus was 5.588, varying from 3 (HTG7) to 10 (ASB17). Average effective number of alleles was 3.603, fluctuating from 1.789 (HMS7) to 5.728 (HMS2). The observed heterozygosity ranged from 0.136 (HMS3) to 0.909 (ASB2) with a mean of 0.631. The results indicate that the studied population originates from the appropriate number of parent generations. The mean expected heterozygosity was 0.690, varying from 0.441 (HMS7) to 0.853 (ASB17) indicating high genetic variability within Bosnian mountain horse population. The PIC values ranged from 0.409 (HMS7) to 0.837 (ASB17) with a mean of 0.643, suggesting that 94.12% markers were quite informative in terms of their suitability for genetic diversity studies .The most polymorphic locus was HMS2 and the least polymorphic locus was HMS7. The inbreeding coefficient ranged from -0.030 (HMS7) to 0.807 (HMS3) with a mean of 0.077. Inbreeding coefficient values indicated no shortage of heterozygotes in Bosnian mountain horses. Deviation from Hardy-Weinberg equilibrium (p<0,05) was found in three loci (HTG10, HMS3 and ASB17). The applied set of 17 microsatellite markers proved to be sufficiently specific for use in genotyping of Bosnian mountain horse. Considering the values of HO, HEand PIC over 0.6, five microsatellite markers system (HTG4, AHT4, AHT5, ASB2, HMS2) is considered to be highly useful for genetic diversity studies in Bosnian mountain horse breed.

Background: The causes of infertility and recurrent spontaneous abortions are diverse and numerous – including non-genetic and genetic factors – whereby the importance of genetic factors in pathogenesis of infertility is becoming more and more common. Chromosomal abnormalities and genetic defects can cause reproduction failures, and for this reason genetic analysis can play an important role in reproductive problems research.Aims and Objective: This study aims to determine the type and frequency of chromosomalaberrations in the female population sample, as well as to determine if the difference between groups with and without chromosomal aberrations was statistically significant.Materials and Methods: One hundred women aged 15-46 were included in the study, allhaving different reproductive disorder diagnoses and requiring karyotype analysis in the Sarajevo Medical Faculty Genetic Center. Cytogenetic analysis was performed on the peripheral blood, which was cultured for four days, using GTG banding forchromosomalanalysis.Results: Out of 100 women included in the study, an abnormal karyotype was found in 16 of them (16%). The difference between the frequency of normal and abnormal karyotype in women with reproductive problems identified in this study was found to be statistically significant. The pattern of chromosomal aberrations was similar to that reported in the previous cytogenetic studies with similar inclusion criteria.Conclusion: This fact should be taken in the consideration in order to estimate true etiology of reproductive problems and it is a valuable information in the process of genetic counseling and decision making in assisted reproductive technology.Asian Journal of Medical Sciences Vol.9(5) 2018 12-16

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 aim of the present study was to investigate values of hematological and some biochemical parameters of autochthonous Bosnian mountain horse breed by revealing the differences related to age and gender. These horses are adapted to specific environmental and climatic conditions in the region and their blood parameters might differ from other horse breeds. Blood samples were collected on stud farm “Borike” from 30 clinically healthy adult horses of both gender (16 females and 14 males), aged between two and twenty-three years. Horses were grouped in three age classes: young mature horses (2-6 years, n=10), middle age horses (7-14 years, n=10) and old horses (>14 years, n=10). A total of eleven hematological and twelve biochemical parameters were analyzed. Gender significantly affected LYM (p=0.009), EOS (p=0.016), UREA (p=0.018), P (p=0.028), TBIL (p=0.048) and AMYL (p=0.033) levels. Values of UREA, TBIL and AMYL were significantly higher in males, while values of LYM, EOS and P were significantly higher in females. Age significantly affected GLU (p=0.019) and UREA (p=0.001) levels. Values of GLU were significantly lower in middle age horses than in old horses while values of UREA were significantly lower in young mature horses than in middle age and old horses. UREA was the only parameter affected by both age and gender. Results obtained in the present study showed that gender had much more powerful effect on the investigated parameters in clinically healthy adult Bosnian mountain horses then did the age.