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Abstract The most commonly farmed fish species in Bosnia and Herzegovina’s aquaculture are from the family Salmonidae, including brook trout Salvelinus fontinalis which is reared both for consumption and stocking purposes. A number of farmers complained about the elevated frequency of anatomical deformities in the smolts and fingerlings of brook trout, decreasing their fitness rate and causing significant financial loss. Since it has been shown that occurrence of deformities is correlated with the low genetic diversity and high inbreeding, this study aimed to assess intra- and interpopulation diversity of Salvelinus fontinalis from different freshwater fish farms in Bosnia and Herzegovina by observing variation in mitochondrial and nuclear genome. Total of 109 samples of brook trout from three hatcheries located at the Neretva River were analyzed for the mitochondrial control region and seven nuclear microsatellite loci. Both PCR-RFLP and sequencing revealed only one haplotype of the control region in all investigated trout. Overall, a low number of genotypes was evident across all the observed loci. Values of genetic diversity and polymorphic information content followed the increase in the number of alleles per locus. In general, values of inbreeding coefficient were generally very high, while the genetic diversity and observed heterozygosity had low rates. The results of our study are congruent with the findings of previous studies in which developmental deformities were concomitant with the low genetic diversity and inbreeding depression. It is, therefore, strongly advised to regularly supplement the broodstock with new, unrelated individuals, as it is of vital importance for sustaining a satisfying level of genetic diversity and preventing inbreeding depression. Additionally, maintaining good management practices regarding the fluctuation of water temperature, exposure to pollution, nutrition, etc., will further contribute to the prevention of this detrimental condition.

A. Mušović, R. Škrijelj, M. Gajević, B. Stroil, A. Vesnić, M. Mitrašinović-Brulić, Samir Đug

Abstract Buško Blato Reservoir, the third largest hydro accumulation in Europe, is situated in the south-eastern part of the karstic Livanjsko Polje Valley. This aquatic ecosystem is distinguished by a very rich ichthyofauna and the presence of four endemic fish species. Numerous studies focus on ichthyological researches of Buško Blato Reservoir. Its ichthyofauna has been exposed to changes in ecological factors, which in turn reflected on the structure and composition of fish populations. The main objective of this paper was to assess the current state and predict future trends in the ichthyofauna structure and dynamics based on the field data and comprehensive analyses of literature data. The results of the research indicated the presence of 11 fish species from four families, which is the largest number of fish species ever recorded in this ecosystem. Sander lucioperca, Lepomis gibbosus, Pseudorasbora parva and Tinca tinca were recorded for the first time in this ecosystem, while some previously recorded species were not found. The results of the analyses clearly indicate the presence of natural interspecific competition and significant level of threats to the endemic fish species caused by human activities.

M. Meland, O. Frøynes, M. F. Akšić, N. Pojskić, B. Stroil, Lejla Lasić, F. Gaši

European plum cultivars (Prunus domestica L.) are hexaploid and partially self-fertile or self-sterile requiring compatible pollinizers with overlapping bloom times. Therefore, inter-planting of different pollinizer cultivars is recommended. In order to identify successful pollinizers of the plum cultivars ‘Edda’, ‘Opal’ (self-fertile), ‘Jubileum’, ‘Reeves’, ‘Mallard’, ‘Avalon’, ‘Cacanska Lepotica’ (self-fertile), and ‘Valor’, 60 fruits per cultivar were collected from nine orchards in 2017 and 2018, all of which were located in Ullensvang, western Norway. DNA extraction was subsequently conducted from the obtained embryos, followed by genetic characterization using seven microsatellite markers. Tissue samples from all possible pollinizers were collected during the summer of 2017 and the same DNA approach was conducted. Results showed that ‘Opal’ was the most successful pollinizer among the investigated plum cultivars. The main exception was ‘Cacanska Lepotica’, which consistently displayed very high level of self-pollination. The most successful foreign pollinizer of ‘Opal’ was ‘Mallard’. However, in more than two thirds of embryos extracted from ‘Opal’ fruits self-fertilization was determined. ‘Reeves’ was identified as the most successful pollinizer among embryos collected from ‘Valor’. Among the five cultivars (‘Edda’, ‘Jubileum’, ‘Reeves’, ‘Mallard’, and ‘Avalon’) that did not display self-pollination, the pollinizer success rate of ‘Opal’, ranged from 36.5% (‘Mallard’) to 93.5% (‘Edda’) in 2017, while in 2018 this rate ranged from 43.5% (‘Jubileum’ and ‘Reeves’) up to 96.5% (‘Edda’). Overall, genotyping embryos using SSRs (simple sequence repeats) proved an effective method in determining the success rate of individual pollinizers among European plum cultivars.

B. Stroil, Lejla Lasić, J. Hanjalić, Sonja Mačar, A. Vesnić

Rhyacophila Pictet 1834 is globally distributed and highly diverse genus of caddisflies (Trichoptera), characterized by numerous regionally endemic species. In the Balkan Peninsula, the highest number of Rhyacophila species (23) was recorded for Bosnia and Herzegovina. Rhyacophila bosnica Schmid, 1970 is found only in the Balkan Dinaric region, with a locus typicus in Vučja Luka, Bosnia and Herzegovina. Like with many species of Trichoptera, the morphology of its larva is still unknown. Therefore, DNA barcoding approach was used to link two developmental stages. In this paper, we report on the first DNA barcode record for this species.

B. Stroil, Semir Doric, J. Hanjalić, Lejla Lasić, N. Pojskić

The expert reports state that Bosnia and Herzegovina, despite the presence of diverse and valuable natural resources, lacks systematic, coordinated and harmonized pipeline for biomonitoring. Successful solutions to serious problems regarding environmental protection, management and research rely on the efficient use of exhaustive and unfailing information on the nature around us. However, more often than not, transitional and developing countries lack any centralized, nationally funded databases that could be used as dependable source of information in decision making process. University of Sarajevo-Institute for Genetic Engineering and Biotechnology (INGEB) developed the Regional Biodiversity Database – REBIDA with the aim to collate all known biological data on wild and domesticated natural resources of Bosnia and Herzegovina. This internet-based database represents a comprehensive, searchable and open access platform for science community, academia, governmental and non-governmental stakeholders and general public. Besides its scientific value, REBIDA will serve as an educational tool for discovering the diversity and importance of natural resources, with special emphasis on indigenous and endemic flora, fungia and fauna from the Balkans. It is the only such database in the country, consisting of three functionally connected segments: tissue database, DNA database and digital genetic database on plant, animal and human samples. To complement REBIDA, a mobile application called REBIDA SCANNER was also developed. It will be free to download for IOS and Android platforms and will enable professionals, nature enthusiasts and any other interested parties to contribute to REBIDA through data collection, field sampling and documentation of B&H wild life.

J. Hanjalić, Semir Doric, Lejla Lasić, B. Stroil, Selma Hasičić, N. Pojskić

The Dinaric endemic plant species Moltkia petraea (Tratt.) Griseb. is often called a "living fossil" of ancient Tertiary flora, with great importance for Bosnia and Herzegovina’s biodiversity. Considering its narrow and limited distribution range, insufficient data on the molecular background of this species is given so far. Due to the presence of various secondary metabolites that interfere with the DNA, isolation of nucleic acids from plant cells is known to be challenging. Even in closely related species it is necessary to optimize DNA isolation protocol in order to obtain high quality PCR amplifiable DNA. We collected 91 samples from five populations in Herzegovina. Doyle and Doyle (1987) CTAB protocol was modified by adding vitamin C (ascorbic acid) to the cell lysis buffer to improve DNA yield and quality. trnL(UAA) intron and nrDNA (ITS1, ITS2) molecular markers were applied to demonstrate amplifiability of isolated DNA and elucidate the intra- and interpopulation genetic diversity. Our results suggest a successful PCR amplification for 81% of the analyzed samples. PCR-RFLP analysis of trnL(UAA) revealed that all individuals in five populations have the same haplotype based on the obtained enzymatic profile for three enzymes (TaqI, HinfI, HindII). Alignment and comparison of ITS sequences didn’t reveal any hypervariable portion that could be informative in elucidating the genetic diversity of M. petraea populations. Further studies with additional application of microsatellite loci, RAPD and AFLP methods are necessary in an attempt to get insights into the genetic diversity of M. petraea.

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.

Cris E. Hughes, Mary P Rogers, A. Owings, Barbara Petzelt, Joycelynn Mitchell, Harold Harry, Theresa Williams, Dena Goldberg et al.

F. Gaši, A. Hodžić, Mirza Hadžiavdić, M. Kurtović, J. Grahić, Lejla Lasić, B. Stroil, N. Pojskić

Raspberry cultivars are clonally propagated and therefore all plants belonging to a single cultivar represent the same genotype. Cultivar integrity of raspberry plantlings placed on the market in Bosnia and Herzegovina (B&H) is based on examining of morphological traits, which is not a reliable tool for genetic identification. In this study plantlings declared as cultivar ‘Polana’ were genotyped using seven microsatellites, in order to gain preliminary insight into the genetic integrity of raspberry plantlings marketed in B&H. Plant tissue (leaves) from 10 raspberry plants were randomly sampled from a batch of plantlings sold by major fruit nursery in Bosnia and Herzegovina. Along with these samples, four reference cultivars with confirmed identity (‘Polka’, ‘Autumn Bliss’, ‘Heritage’ and ‘Polana’) were also included in the study. Seven primer pairs amplified 31 alleles, or on average 4.4 alleles per locus. UPGMA cluster analysis, based on the Jaccard similarity coefficient, revealed that among the ten samples declared as ‘Polana’ plantlings only five were genetically identical to any of the other samples. The cluster analyses also exposed that none of the ten samples declared as ‘Polana’ seedlings were in fact identical or even closely related to the ‘Polana’ reference cultivar or any of the other reference cultivars. These findings clearly show that the genetic identity of primocane raspberry plantlings , currently sold in Bosnia and Herzegovina, needs to be tested using objective and reliable methods rather than simple morphologic observation.

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