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Dunja Rukavina

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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.

M. Mačkić-Đurović, Amina Aščerić, D. Rukavina, I. Aganović-Mušinović

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.

Mirela Mackić-Durović, D. Rukavina

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.

M. Mačkić-Đurović, Nejla Đikić, D. Rukavina

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.

Amela Bajri̇ć, M. Kati̇ca, A. Katica, Alisa Smajović, Mujo Sivić, D. Rukavina, Mirza Čelebičić, A. Zahirović, Ajla Bajrić et al.

Introduction: The technique of closing the appendiceal stump using linear titanium stapler clips is being used more and more frequently in laparoscopic surgery, despite the good practice in the use of resorbable single endoloop vicryl ligatures and/or the non-resorbable plastic hem-o-lok clips. No light has been shed on potential undesirable effects on the haemathological profile of the use of titanium stapler clips. This experimental study aimed at establishing any possible undesirable effect of linear titanium stapler clips and single resorbable vicryl endoloop ligatures on the blood cells in rats following appendectomy. Methods: A total of 70 adult rats were used, divided into a control group (n = 10), and two experimental groups (n = 30 + 30). The appendices were removed from the rats in the first experimental group using titanium stapler clips and in the second experimental group the appendiceal stump using resorbable endoloop vicryl ligatures was closed. In both experimental groups three sub-groups with ten animals each were formed, from which peripheral blood from a tail vein on days 7, 28 and 60 was removed postoperatively, in order to assess the usual haematological parameters. Stained blood smears were also analysed in order to establish any poikilocytotic erythrocytes present. Results: In the first experimental group, with the titanium stapler clips, more than 25 % neutrophils were found on day 7, which is a significantly different result (p < 0.05) to the control group. In the second experimental group, there were more neutrophils than in the titanium stapler clips group, especially on days 7 and 60 and the results of these two sub-groups differ statistically significantly, p < 0.05. Hypochromia was found in the endoloop vicryl ligatures group, as well as in the titanium stapler clips sub-group on day 28, due to lower haemoglobin values which were significantly different to the control group, p < 0.05. Moderate levels of annulocytes, spherocytes and stomatocytes were found in most experimental groups. Conclusion: The results of this study favour the use of linear titanium stapler clips over resorbable single endoloop vicryl ligatures, because a less unfavourable effect was established on the blood cells of the experimental rats with their use.

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.

D. Rukavina, I. Alilović, Ć. Crnkić, A. Ajanović, D. Preldžić, A. Čoralić, D. Čamo, D. HADŽIJUNUZOVIĆ-ALAGIĆ, Selma Filipović et al.

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