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Adaleta Durmić-Pašić

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Bosnia and Herzegovina (B&H) is among the European countries with the highest rate of air pollution-related death cases and the poorest air quality. The main causes are solid fuel consumption, traffic, and the poorly developed or implemented air pollution reduction policies. In addition, the city of Sarajevo, the capital of B&H, suffers temperature inversion episodes in autumn/winter months, which sustain air pollution. Human biomonitoring studies may be confounded by the lifestyle of subjects or possible metabolic alterations. Therefore, this study aimed to evaluate Ligustrum vulgare L. as a model for air pollution monitoring by measuring DNA damage at one rural and two urban sites. DNA damage was measured as tail intensity (TI) in L. vulgare leaves, considering seasonal, sampling period, leaf position and staging, and spatial (urban versus rural) variation. Effects of COVID-19 lockdown on TI were assessed by periodical monitoring at one of the selected sites, while in-house grown L. vulgare plants were used to test differences between outdoor and indoor air pollution effects for the same sampling period. Significantly higher TI was generally observed in leaves collected in Campus in December 2020 and 2021 compared with March (P < 0.0001). Outer and adult leaves showed higher TI values, except for the rural site where no differences for these categories were found. Leaves collected in the proximity of the intensive traffic showed significantly higher TI values (P < 0.001), regardless of the sampling period and the stage of growth. In regards to the COVID-19 lockdown, higher TI (P < 0.001) was registered in December 2020, after the lockdown period, than in periods before COVID-19 outbreak or immediately after the lockdown in 2020. This also reflects mild air pollution conditions in summer. TI values for the in-house grown leaves were significantly lower compared to those in situ. Results showed that L. vulgare may present a consistent model for the air pollution biomonitoring but further studies are needed to establish the best association between L. vulgare physiology, air quality data, and air pollution effects.

A. Ahatović, E. Al‐Momani, K. Bajrovic, A. Durmić-Pašić

Since the commercialization of transgenic crops in 1996, the biotech crop planted area has continuously increased. The European consumers have particularly been sceptical about transgenes in food products and EU (European Union) has enacted very complex legislation. The area of food analytics requires continuous development and improvement of detection methods to track the legislative framework and respond to consumers requirements. In the last decade, real-time PCR (polymerase chain reaction) based methods have been the methods of choice for numerous laboratories, but for various reasons, end-point PCR based methods have still been used. In our research, 73 samples of food and feed were analysed for the presence of common elements of transgene construct – Cauliflower Mosaic Virus 35S promoter (P-35S) and Agrobacterium tumefaciens Nopaline Synthase Terminator (T-NOS), using end-point PCR based methods. These samples had been previously tested for the presence of the same elements using validated real-time PCR based methods. Comparison of the used methods sensitivity showed that real-time PCR based methods have undeniable advantage. More important factor is specificity, and the fact that the list of approved Genetically Modified Organisms (GMOs) is constantly increasing necessitates updating of validation methods procedures. Considering upward trend of approved GMOs, it is important to pay more attention to the improvement and specialization of GMO detection methods.

Of the four species of the genus Satureja (Lamiaceae) that are recognized in Bosnia and Herzegovina, S. subspicata has the the widest distribution. It is taxonomically challenging species of geographically limited distribution and little data on its genetic diversity throughout its range is available. We sampled six geographically distinct populations from Bosnia and Herzegovina and applied nrDNA (ITS1, ITS2), chloroplast markers (matK and trnL) and AFLP to examine genetic diversity of S. subspicata in the center of its distribution range and to explore the possibility of establishing the species DNA barcode. AFLP analysis showed large genetic differentiation among populations as well as moderate correlation between genetic distance among populations and geographic distance among locations. MatK has not proven useful in distinguishing S. subspicata from sympatric species. However, nrDNA sequences provided necessary resolution power, with ITS2 being more informative. Estimates of evolutionary divergence between nrDNA sequences obtained in our research and homologous sequences of sympatric Satureja deposited in the GenBank reveal closer relationship between geographically proximate populations of different species and slight divergence within S. subspicata sequences pool. This outcome highlights the importance of considering overall genetic diversity across the distribution range of a species when assigning DNA barcode.

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.

A. Ahatović, I. Brkic, Lejla Cokoja, Azra Muhovic, A. Durmić-Pašić

Following the “horse meat scandal” in 2013, European Union countries have conducted official control of EU market and unraveled food fraud which implicated a number of processed food products and food businesses. Five years after the breakout of the scandal, no official information on market surveillance in Bosnia and Herzegovina is available. Therefore, 73 randomly selected meat products from retail were collected and analyzed for the presence of horse DNA. Horse DNA was detected in 21 products (28.77%). Particularly disturbing for B&H consumers is high proportion of sujuk samples positive for horse DNA (46.15%) with lower incidence among the products of small manufactures. Also disturbing is the finding that 71.43% of the products that contain horse DNA were produced in B&H. According to our data there is a requirement for stricter surveillance of both import and internal market.

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