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.
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.
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.
Apart from its physiological role in the cellular oxidation of ethanol interesting feature of the ADH1B gene locus is its characteristic geographical distribution in which certain variants of ADH1B peak in different parts of the world. Therefore, ADH1B rs2066701 polymorphism is exploited as a genetic marker in tracing of the evolutionary processes and human migrations in the past. Taking into consideration the complexity of population genetic structure and several migrations in the history of the Balkan populations, including Bosnian and Herzegovinian, this study aimed to estimate the frequency of ADH1B rs2066701 polymorphism in the population of Bosnia and Herzegovina. The total of 101 randomly sampled individuals was genotyped for rs2066701 polymorphism in ADH1B gene using PCR-RFLP method. The obtained frequencies were used to calculate heterozygosity, fixation indices and Hardy-Weinberg equilibrium. Observed population-structure parameters were compared with other population values available in ALFRED database. Dimensional relations between the investigated populations were visualised with the NM-MDS (non metric multidimensional scaling) analysis using PAST. The minor allele frequency for rs2066701 was 0,257. Inter-population analysis including other European and non-European populations from the ALFRED database proved the above-mentioned European genetic background of the B&H population.
Abstract Innovation holds the potential for economic prosperity. Biotechnology (BT) has proved to be a viable vehicle for the development and utilization of technologies, which has brought not only advances to society, but also career opportunities to nation-states that have enabling conditions. In this review, we assess the current state of BT-related activities within selected new and preaccession EU countries (NPA) of CEE region namely Croatia, Romania, Bosnia and Herzegovina and Serbia, examining educational programs, research activity, enterprises, and the financing systems. The field of BT covers a broad area of activities, including medical, food and agriculture, aquaculture or marine, environmental, biofuels, bioinformatics, and many others. Under the European Commission (EC), member-states are to set their Research and Innovation Strategies for Smart Specialization (RIS3), to identify priorities or strengths in order to develop knowledge intensive economies. As the four countries highlighted in this review are in the early stages of implementing RIS3 or have not yet fully formulated, it presents an opportunity to learn from the successes and failures of those that have already received major structural funds from the EC. A critical point will be the ability of the public and private sectors’ actors to align, in the implementation of RIS3 as new investment instruments emerge, and to concentrate efforts on a few select target goals, rather than distribute funding widely without respect to a long-term vision.
The second most dominant genetically modified (GM) crop is maize. Increasing number of GM maize events puts significant pressure on GMO testing laboratories to achieve the level of competence necessary to fulfill legal requirements. In the European Union, Polymerase Chain Reaction (PCR) is the method of choice for identification and quantification of GMOs. We performed verification of validated methods for identification of four GM maize events. Additionaly we aimed to explore the option of designing a method for simultaneous detection of these events in a multiplex PCR reaction. DNA was extracted from certified reference materials (CRM) using validated CTAB extraction protocol. Concentration of DNA was measured using Qubit dsDNA Broad Range Assay. Amplification of taxon specific marker for maize and all event-specific methods was performed according to the JRC Compendium of Reference Methods for GMO Analysis. Absolute limit of detection (LODabs) was determined for taxon specific and four event specific RealTime PCR based methods. DNA extracted from CRMs showed sufficient concentration for downstream analyses and preparation of dilutions for determination of LODabs. Determined LODabs for all tested methods meet acceptance criteria. As expected, the methods performance with respect to the repeatability and precision decline with the decrease in concentration of the target. Event-specific GA21 and NK603 methods show high Ct values at the determined LODabs. However, by adjusting the concentrations of primers and probes sensitivity of these two methods should be improved. Considering that the amplicons for all five methods are quite short (<120 bp) optimization of multiplex reaction conditions for simultaneous amplification should be feasible
Soy sauce is worldwide popular condiment of Asian origin. With the advent of GM soybean production, soy sauce drew the interest of food safety control. Samples collected for inspection are generally of industrial grade soy sauce type, which is produced from hydrolyzed soybean and grain. Following the failure to perform RealTime PCR based GMO screening on a number of submitted samples we tested our screening system on soy sauce produced following traditional method based on fermentation. Four batches of soy sauce were produced and DNA extracted. DNA concentration ranged from 32,68 to 65,36 ng/μl. Amplification of taxon specific target was successful with rather high Ct ( > 30). Promoter P-35S sequence was not detected, but T-NOS was detected in three samples with values reaching or exceeding LOD of the method. The results show that it is possible to detect transgenic elements in traditionally produced soy sauce while DNA extraction from industrial grade soy sauce is not possible.
Apart from its physiological role in the cellular oxidation of ethanol an interesting feature of ADH1B gene is its characteristic geographical distribution where certain variants of ADH1B peak in different parts of the world. Therefore, ADH1B rs2066701 polymorphism is used as a genetic marker in tracing the evolutionary processes and human migrations over time. Taking into consideration the complexity of population genetic structure and a number of migration events in the history of the Balkan populations this study aimed to estimate the frequency of ADH1B rs2066701 polymorphism in the population of Bosnia and Herzegovina. The total of 101 randomly sampled individuals were genotyped for rs2066701 polymorphism in ADH1B gene using PCR-RFLP method. The obtained frequencies were used to calculate heterozygosity, fixation indices and HardyWeinberg equilibrium. The observed population-structure parameters were compared with other population values available in ALFRED database. Dimensional relations between the investigated populations were visualised with the NM-MDS (non metric multidimensional scaling) analysis using PAST. The minor allele frequency for rs2066701 was 0,257. Inter-population analysis including other European and non-European populations from ALFRED database proved the above-mentioned European genetic background of the B&H population.
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