AimCOVID-19 pandemic, caused by SARS-CoV-2, has had a profound impact on global health, including in Bosnia and Herzegovina, which faced unique challenges due to limited testing and high mortality rates. This analysis aimed to identify mutations and detect different SARS-CoV-2 lineages across four pandemic waves.MethodologyA total of 127 SARS-CoV-2 samples were collected and sequenced from patients from the Federation of Bosnia and Herzegovina, providing a comprehensive overview of the viral genetic diversity in this region. Two sequencing platforms, Ion Torrent and Illumina, were used, whereby 37 samples were sequenced on the Ion Torrent platform, while others were sequenced on the Illumina platform.ResultsThis study presents a genomic analysis of SARS-CoV-2 variants circulating in the Federation of Bosnia and Herzegovina over four distinct pandemic waves, spanning from March 2020 to April 2023. Examination of genomic variations across these waves revealed key mutations associated with transmission and potential virulence.ConclusionThese genomic insights into SARS-CoV-2 evolution in Federation of Bosnia and Herzegovina emphasizes the importance of continuous surveillance to understand viral evolution and strengthen public health responses to future pandemics.

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.

Isolation-by-distance (IBD) pattern among bilberry (Vaccinium myrtillus L.) populations has previously been reported for this species in northern Europe. However, the number of molecular studies conducted on bilberry, using everything from isoenzymes, RAPDs to microsatellite markers, are very few and far between. Considering that Bosnia and Herzegovina (B&H) is a country rich with diverse fruit genetic resources, conducting a genetic characterization of the naturally occurring V. myrtillus populations could yield valuable data for the conservation and utilization of this resource. This study entailed genotyping samples collected from three bilberry populations located in Fojnica, Kladanj, and Srebrenica municipalities using seven polymorphic microsatellite or SSR (simple sequence repeats) markers. The obtained molecular data was used to calculate the correlation between the physical distance of the individual B&H populations and a parameter of the genetic differentiation (pairwise Fst). The results of the correlation analyses revealed an absence of a significant isolation-by-distance pattern among the three examined B&H bilberry populations. In addition, the most pronounced genetic differentiation was detected between the Srebrenica and each of the two remaining B&H populations. At the same time, the values for pFst were significant, albeit much lower, between the Fojnica and Kladanj populations. Bilberries from the sampled Srebrenica population appear to be distinct from the other B&H populations, possibly due to the different genetic origin of this population.

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.

This study is the first report on the mtDNA profile of human settlements of the Konjuh and Majevica mountains of northeastern Bosnia. The aims of this study were: a) determination of mitochondrial genetic structure of populations of the Konjuh and Majevica mountains of northeastern Bosnia; b) detection of trace of ancient of mtDNA variations; and c) assessment of genetic relations with other Bosnian and Herzegovina populations and neighboring populations from the Balkan region. The genetic structure of populations of Konjuh and Majevica is shaped by western Eurasian maternal signals, which may trace their ancestry to the Paleolithic, pre-Neolithic and Neolithic. Especially interesting is the feature of the Neolithic expansion in this area. This applies especially to the presence of the pre-Neolithic lineages HV*and N1a in northeastern Bosnia, which can indicate an early settlement of this region of Bosnia by pre-Neolithic populations from the Middle East. This region abounds with resources of salt sites, which might suggest in favor of the thesis that the early-Neolithic colonists needed a safe source of salts so as to settle in the Balkan area. The populations of mountains of northeastern Bosnia indicate elements of the local population history, but they do not show strict genetic closure in relation to the neighboring populations of the Balkans. This may be a consequence of the population size, degree of geographic isolation and events of migration.

Abstract Background: Low-grade chronic inflammation is an important feature of chronic kidney disease (CKD). Aim: To determine the values of C-reactive protein (CRP), neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) in patients with different stages of CKD and to examine how they change depending on the progression of renal damage. Materials and methods: A cross-sectional descriptive comparative study included 157 subjects at different stages of CKD which was assessed based on glomerular filtration rate (GFR) calculated according to the MDRD equation. CRP was analyzed by an immunoturbidimetric method. NLR and PLR were calculated by a mathematical calculation after a blood count was performed. Results: The present study showed an increase in serum creatinine, CRP, and NLR values with progression of renal failure. There was a statistically significant difference in the creatinine and CRP concentrations between groups with different stages of CKD (p <0.001 for all comparisons). A significant positive correlation was found between NLR and CRP, while negative, significant correlations were observed between NLR and eGFR as well as between PLR and eGFR. There was a slight increase in PLR value with the progression of renal impairment, but the correlation between PLR and CRP was not significant. Conclusion: These results suggest that NLR, together with CRP, may serve as an indicator of systemic low-grade inflammation progression in patients with CKD. Larger prospective studies are required to observe the possibility of using NLR as a surrogate marker for CRP in patients with CKD.

ABSTRACT Population genetic studies have shown that the Bosnian-Herzegovinian (B&H) population is a part of the European gene pool, but there has been limited information on the genetic structure of ancient B&H populations. This study aimed to determine the frequency and distribution of mitochondrial DNA (mtDNA) haplogroups for a medieval Bosnian population. Thirty-four samples, excavated from medieval necropolises located within the borders of medieval Bosnia, were analyzed. Sequencing of the mtDNA hypervariable segment 1 (HVS1) region and RFLP analysis were performed for haplogroup determination. All 32 samples were identified as haplogroup H, with subhaplogroups H2a and H5 in 30 and 2 samples, respectively. The frequency of the H haplogroup was significantly different between the studied samples and previous studies of contemporary B&H populations, where the H haplogroup frequency was approximately half that of the ancient population studied here. A significant difference in H haplogroup frequency compared with other medieval populations outside of Bosnia was also observed: the ancient B&H population is most similar to ancient Italians. These results provide insight into the mitochondrial landscape of populations that inhabited the territory of present-day Bosnia and Herzegovina in the Middle Ages. Our study reveals that inhabitants of medieval Bosnia carried genetic lineages that exist today in B&H populations, suggesting continuity of mtDNA haplogroups over a long period of time, regardless of various historical demographic events that shaped the genetic structure of the modern B&H population.

In order to best conserve, as well as utilize, traditional apple germplasm in Norway, an apple heritage cultivar collection was established in Ullensvang, western Norway, which aims to become the National Clonal Germplasm Repository. The establishment of the apple heritage cultivar collection was preceded by a molecular study that aimed to genotype a large number of apple accessions maintained in various ex situ sites in western and south-eastern Norway, using a rather small set of eight SSR markers. However limited, the marker set managed to identify synonyms, homonyms, and duplicates within and among the investigated collections. In this study, 171 apple accessions from the Ullensvang apple heritage cultivar collection were genotyped using a set of 20 different SSR markers. Approximately half of the accessions have been previously genotyped using eight SSR markers, enabling an assessment of whether the use of a larger marker set would yield a more accurate characterization. Based on the obtained molecular data, the apple heritage cultivar collection was determined to hold a key part of the overall genetic diversity of the Norwegian apple germplasm. Furthermore, the twelve additional SSR markers were able to differentiate several accessions groups originally thought to be synonyms, as well as to provide a more detailed insight into the genetic structure of this germplasm.

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.