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Lejla Usanovic

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Background and purpose: Three genera and eleven species of Glossosomatidae family have been confirmed in Bosnia and Herzegovina. However, there is limited research on the biodiversity of this family in the region, especially on molecular data. With this paper, we contribute to the knowledge on the Glossosomatidae family with the first molecular records for two species for Bosnia and Herzegovina. Materials and methods: Larval samples of caddisflies were collected in April 2019 and 2021 at the area of the Natural Monument the Spring of Bosna River followed by morphological identification. Molecular confirmation of species was done using DNA barcoding, targeting the cytochrome c oxidase subunit I (COI) region using Folmer’s primers and degenerated primers LCO1490-JJ and HCO2198-JJ. All obtained PCR products were bidirectionally sequenced using the same primers as in PCR. Results: Out of 52 specimens, 18 were morphologically determined to species level. A DNA barcode was successfully generated for 16 specimens. Eight sequences were generated for Synagapetus krawanyi, revealing three haplotypes previously absent from the BOLD database. Four sequences were obtained for S. iridipennis, with one new haplotype for BOLD database. Conclusions: We present new data on species distribution and genetic diversity of S. krawanyi and S. iridipennis from Bosnia and Herzegovina. This study contributes to the knowledge of caddisfly biodiversity and the Global Trichoptera Barcode Initiative by adding new records from two underrepresented species in the BOLD database.

Polyploidy is a powerful mechanism driving genetic, physiological, and phenotypic changes among cytotypes of the same species across both large and small geographic scales. These changes can significantly shape population structure and increase the evolutionary and adaptation potential of cytotypes. Alyssum moellendorfianum, an edaphic steno-endemic species with a narrow distribution in the Balkan Peninsula, serves as an intriguing case study. We conducted a comprehensive analysis of genetic diversity and population structure across the species’ range, employing an array of genetic techniques (nuclear microsatellites, amplified fragment length polymorphisms, and plastid DNA sequences), flow cytometry (FCM), morphometry, and pollen analysis. The study reveals two genetic lineages: spatially distributed diploid and tetraploid cytotypes. Clear divergence between diploids and tetraploids was shown by AFLP, while plastid DNA sequences confirmed private haplotypes in each of the studied populations. Higher genetic diversity and allelic richness following the north-south pattern were documented in tetraploids compared to diploids, as indicated by nuclear microsatellites. Morphometric analysis via principal component analysis (PCA) and canonical discriminant analysis (CDA) did not reveal any divergence between diploid and tetraploid cytotypes. Nonetheless, a distinction in pollen size was clearly observed. The results suggest an autopolyploid origin of tetraploids from diploid ancestors. Despite the population fragmentation in a very small geographic range, these populations harbour high genetic diversity, which would allow them to remain stable if natural processes remain undisturbed.

ABSTRACT Borrelia burgdorferi sensu lato complex comprises 20 species, from which B. afzelii, B. garinii, B. burgdorferi sensu stricto, B. bavariensis, and B. spielmanii are directly associated with Lyme borreliosis, while B. bissettiae, B. lusitaniae, and B. valaisiana were detected in individual cases. Their main vector in Europe is the hard tick species Ixodes ricinus. To date, two species, B. spielmanii and B. lusitaniae, have been molecularly detected in Bosnia and Herzegovina. To test for the presence of other Borrelia species, we performed nested PCR targeting intergenic region rrf (5S) – rrl (23S) on DNA isolates from 49 ticks collected from vegetation by flagging method and 43 removed from humans in The Center for Emergency Medical Assistance of the Sarajevo Canton and The Healthcare Centers of the Sarajevo Canton. Borrelia species were detected by one-directional Sanger sequencing of the amplified region using the same forward primer as in PCR. Out of six Borrelia species detected in the present study, this is the first record of B. afzelii, B. garinii, B. burgdorferi s.s. B. bavariensis, and B. valaisiana in Bosnia and Herzegovina.

Background and purpose: Available data in research literature suggest that the Western Balkan countries hold a rich diversity of caddisflies. Assessment and biomonitoring of such rich diversity could be facilitated through DNA-based high-throughput approaches like DNA metabarcoding that depend on the availability of comprehensive reference libraries. Materials and methods: We assessed the status of the COI barcode sequence data for a total of 112 caddisflies species in the investigated region by determining the gaps in representative sequences in the Barcode of Life Data System (BOLD) and examining the accuracy of available records using the Barcode, Audit and Grade System (BAGS). Results: Results revealed a considerable underrepresentation of surveyed geographic region in BOLD records for the target insect group. Moreover, the large majority of the species records were rated “discordant” (72.80% grade E), and only 15.20% were classified as “consolidated concordance or basal concordance” (3.20% grade A and 12.00% B). Approximately 3.20% of the records pertaining to species occurring in multiple BINs (Barcode Index Number) and 8.80% were poorly represented (i.e., less than three specimens, grade D). A fraction of the species graded discordant were deemed concordant after detailed inspection of individual data, decreasing by 14.07%. Conclusions: The assessment of the current state of BOLD entries indicated that DNA barcoding is still not widely applied in Albania, Bosnia and Herzegovina, Montenegro, North Macedonia, Serbia, and Slovenia, emphasizing that Croatia has the most barcoded caddisflies species. The finding that available BOLD Trichopteran records for investigated countries were mainly graded as “discordant” indicates the need for better quality control of reference libraries.

Introduction: The COVID-19 pandemic and the restrictions from routine life habits had a tremendous impact on psychological and physical health of youth. It is known that stress, anxiety and depression can be associated with the development of gastrointestinal (GI) symptoms and known to exacerbate present GI symptoms. The pandemic has forced many changes in the behavior of student population such as the studying in an asocial environment. The aim of this study was to examine and quantify the influence of stress onto the quality of life and GI symptoms in the student population in Bosnia and Herzegovina (B&H), before and during the pandemic lockdown. Methods: A total of 279 students from B&H were assessed for their GI and emotional status in pre-COVID period and during the COVID period using validated instruments: GI symptom rating scale (GSRS), Visceral Sensitivity Index, and the Patient Health Questionnaire 15-item Somatic Symptom Severity Scale. Results: The results showed that moderate and severe GI symptoms were more frequently present among student population at the time of the pandemic than in period before pandemic. The most pronounced symptoms were bloating syndrome and abdominal pain syndrome according to the GSRS. Conclusions: We concluded that concern for one’s health and changed way of life are directly related to a worsening of the symptoms of GI disorders in the student population. Further research should go in the direction of early prevention of GI disorders that take root in early youth and later develop into chronic forms.

Silene sendtneri Boiss. (Caryophyllaceae) is the Dinaric endemic plant species with white, decorative and scented flowers. Previous studies on this endemic species were based on morphology and effects on seed germination after the treatment with salicylic acid. However, no molecular genetic studies have been conducted on this species so far. This paper presents preliminary results of the usefulness of microsatellite loci created for cosmopolitan species in assessing the genetic diversity of endemic plant species. A total of 100 specimens were collected from 18 localities in the mountain regions of Treskavica, Igman, Bjelašnica and Ozren in Bosnia and Herzegovina. No S. sendtneri individuals were found at the mountain Trebević. We tested cross-amplification success and a polymorphism level for the set of microsatellite markers (Sil01, Sil03, Sil16, Sil31, Sil35) designed for the cosmopolitan species Silene nutans. In 100 analyzed individuals of S. sendtneri, Sil31 and Sil35 did not amplify, Sil01 was monomorphic and the remaining two loci showed a high level of allelic diversity. Our findings suggest that caution should therefore be exercised in selecting microsatellite markers designed for cosmopolitan plant species in the analyses of endemic species of the same genus since different genetic factors affect the amplification success and polymorphism of the given loci. Attention should be given to the number of detected and effective alleles and their ratio, the success of locus amplification concerning the complete set of markers used, and the ratio of polymorphs to the total number of observed loci.

Abstract Interspecific hybridization in the Cyprinidae family has been recorded worldwide, with Abramis brama (bream) and Rutilus rutilus (roach) as one of the often-reported hybridizing pairs. The only account of such an event in Bosnia and Herzegovina has been in Modrac Reservoir. Using morphological and molecular markers, the presence of hybrids was surveyed, the hybridization direction was determined and the hybrid group structure in this ecosystem was evaluated. Our findings confirmed unhindered natural hybridization between roach and bream in Modrac Reservoir. Over 50% of the hybrid specimens were classified as F2 hybrids by the NewHybrids software, while the rest were categorized as pure parental form, making it the first such finding in Europe. The analysis of mitochondrial cytochrome b showed that 90% of hybrid individuals were of bream maternal origin. The hybrid group expressed higher mean values of observed heterozygosity and gene diversity than both parental species. Signs of introgressive hybridization between parental species were detected. The hybrid zone of Modrac Reservoir appears to follow the intermediate or “flat” hybrid model based on the balanced distribution of parental and hybrid genotypes. Further investigation is needed to elucidate the factors that enable the survival and mating success of post-F1 individuals.

Lejla Ušanović, J. Hanjalić, Semir Doric, Lejla Lasić, Jasmina Šubara, Adnan Čučuković, B. Kalamujić-Stroil

UDK: 577.212:574]:007.5(497.6) DNA barcoding is a method designed to provide rapid and precise species identifications by using one or more of short gene sequences called barcodes. In most plant and fungi studies, the standard barcodes of choice are three plastid (rbcL, matK and trnH-psbA) and one nuclear (ITS) gene regions. The relatively high, but comparatively conserved rate of sequence evolution of mtDNA has made COI the marker of choice in animals. BOLD is a freely available cloud-based data storage and analysis platform developed with the aim to advance biodiversity science through DNA barcoding species identification. To date, over 6 million barcodes have been deposited in BOLD with 196,000 animal species, 68,000 plant species and 22,000 species of fungi and other organism entries. In this database, there are currently 447 entries for organisms from Bosnia and Herzegovina, which makes 0.0067% of the total number of BOLD entries. According to BOLD statistics, only 1.11% of all organism entries from B&H were submitted by B&H institutions. Despite the fact that Bosnia and Herzegovina has valuable natural resources with a high percentage of endemic and autochthonous species, BOLD statistics elucidated the lack of coordinated and systematic DNA barcoding research so far. It is necessary to establish continuous progress of molecular-genetic characterization of these resources in the future. It is up to B&H institutions to decide if they want to continue the practice of submitting the data sporadically or if they will animate the research community to actively participate in this global project.

M. Piria, B. Stroil, D. Giannetto, A. Tarkan, A. Gavrilovic, Ivana Špelić, Tena Radočaj, N. Killi, H. Filiz et al.

Abstract. Alongside climate change, the introduction of non-native species (NNS) is widely recognized as one of the main threats to aquatic biodiversity and human wellbeing. Non-native species and biodiversity are generally low priority issues on the political agendas of many countries, particularly in European countries outside the European Union (EU). The objectives and tasks of this study were to address the policy regulation, education level, education practices, and socioeconomic perceptions of NNS in the Balkans. A questionnaire-based survey was conducted in Albania, Bosnia and Herzegovina, Montenegro, North Macedonia and Turkey (Balkan EU candidate and potential candidate members), in Croatia and Greece (Balkan EU Member States) and Italy (non-Balkan EU Member State). The EU Alien Regulation (1143/2014) concerning NNS is implemented in EU Member States and Montenegro, whereas Albania, Bosnia and Herzegovina and Turkey have not reported specific policy regulations for NNS. Permanent monitoring programmes specifically designed for NNS have not yet been established in the EU Member States. Most countries tackle the issue of NNS through educational activities as part of specific projects. Education level is indicative of the implementation of NNS policy regulation, and efforts are needed for the proper development of relative study programmes. Public awareness and educational preparedness concerning NNS in the Balkans were identified as poor. Strong programmes for management and education should be developed to increase public awareness to prevent further biodiversity losses in the Balkan region.

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