The first molecular record of Borrelia afzelii, B. garinii, B. valaisiana, B. burgdorferi s.s. and B. bavariensis in Bosnia and Herzegovina.

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.

The Diatom Diversity and Ecological Status of a Tufa-Depositing River through eDNA Metabarcoding vs. a Morphological Approach-A Case Study of the Una River (Bosnia and Herzegovina).

Tufa deposits in karst rivers are unique habitats created by mutual interactions between specific environmental and biotope features and inhabited by diatoms as a highly abundant and diverse algal group. This pilot study aimed to investigate the diversity of diatom communities on tufa depositing habitats and assess the Una River's ecological status using a comparative molecular and morphological approach for diatom identification. The 312 base pairs of the rbcL gene were barcoded and analyzed using MiSeq reads and amplicon sequence variants (ASVs) obtained by the DADA2 pipeline. The reference database Diat.barcode v7 was used for taxonomic assignment. The morphological identification of the diatoms was carried out in parallel. In total, the combined dataset revealed 46 taxa identified at genus rank, 125 on the subgenus, and 145 on combined taxonomy rank. The metabarcoding approach mostly leads to a lower number of identified taxa at species rank (58 in molecular vs. 119 in optical inventory), resulting in higher values of beta diversity and heterogeneity in diatom assemblages in samples obtained by morphological approach. Despite the high percentage of taxonomically not assigned diatom ASVs to the species rank, high Shannon diversity index values and a similar number of taxa per locations compared to the morphological approach were obtained. Taxa Achnanthidium minutissimum (Kützing) Czarnecki, Achnanthidium pyrenaicum (Hustedt) H.Kobayasi, Amphora pediculus (Kützing) Grunow, Diatoma vulgaris Bory, Navicula cryptotenella Lange-Bertalot, and Navicula tripunctata (O.F.Müller) Bory were identified at all locations in both inventories. Although limited consistency in the diatom abundances between the two inventory datasets was found, a similar grouping of samples was observed connected to the river's longitudinal gradient. The data obtained using molecular approach in most sites indicated a mostly lower ecological status (good or moderate) compared to the data obtained from the morphological approach (high, good, and moderate). The potential of environmental DNA (eDNA) diatom metabarcoding for water monitoring and diversity studies is undeniable, but to fully realize the benefits of these methods in the future, it is essential to standardize protocols and expand the reference database for species found in specific habitats, such as tufa deposits.

Monitoring of species' genetic diversity in Europe varies greatly and overlooks potential climate change impacts.

Genetic monitoring of populations currently attracts interest in the context of the Convention on Biological Diversity but needs long-term planning and investments. However, genetic diversity has been largely neglected in biodiversity monitoring, and when addressed, it is treated separately, detached from other conservation issues, such as habitat alteration due to climate change. We report an accounting of efforts to monitor population genetic diversity in Europe (genetic monitoring effort, GME), the evaluation of which can help guide future capacity building and collaboration towards areas most in need of expanded monitoring. Overlaying GME with areas where the ranges of selected species of conservation interest approach current and future climate niche limits helps identify whether GME coincides with anticipated climate change effects on biodiversity. Our analysis suggests that country area, financial resources and conservation policy influence GME, high values of which only partially match species' joint patterns of limits to suitable climatic conditions. Populations at trailing climatic niche margins probably hold genetic diversity that is important for adaptation to changing climate. Our results illuminate the need in Europe for expanded investment in genetic monitoring across climate gradients occupied by focal species, a need arguably greatest in southeastern European countries. This need could be met in part by expanding the European Union's Birds and Habitats Directives to fully address the conservation and monitoring of genetic diversity.

Genetic diversity of Banat Naked Neck, indigenous chicken breed from Serbia, inferred from mitochondrial DNA D-loop sequence and microsatellite markers.

Banat Naked Neck is the most important indigenous breed of chickens in Serbia. Marginalized until recently, it is becoming increasingly popular due to its adaptability and good productivity in alternative production systems. However, its history and the current breeding model pose challenges for breed preservation and future improvement. This study aimed to assess the genetic diversity and structure of four subpopulations of Banat Naked Neck from different districts in Serbia (West Backa, North Banat, South Banat and Kolubara) using D-loop mitochondrial DNA sequences and a set of 30 microsatellite markers. Seven haplotypes in the phylogenetic analysis of D-loop mitochondrial DNA suggested maternal origin related to the Indian subcontinent, while haplotype and nucleotide diversity averaged 0.731 ± 0.053 and 0.0067 ± 0.0018, respectively. Microsatellite genotyping showed an average detected number of alleles per locus of 5.129 ± 0.237, while the observed and expected heterozygosity averaged 0.560 ± 0.018 and 0.631 ± 0.014, respectively. Genetic differentiation estimated through FST was 0.051 (p < .001). Two clusters in STRUCTURE analysis showed possible separation of two older subpopulations (South Banat and Kolubara) from the two more recent ones (West Backa and North Banat). This first comprehensive study of genetic diversity serves as the basis for future preservation, use and improvement of the Banat Naked Neck breed.

Molecular-genetic diversity of the endangered Dalmatian barbelgudgeon, Aulopyge huegelii from the Busko Blato reservoir.

A number of studies investigating different aspects of IUCN endangered species, Aulopyge huegelii Heckel, 1843 (Dalmatian barbelgudgeon) biology have been conducted, but data on molecular genetics are lacking. The goal of this survey was to assess the genetic structure of the A. huegelii population from the Busko Blato reservoir, based on four mitochondrial DNA regions and five microsatellite loci. Excluding cytochrome b, more than one haplotype has been detected in all sequenced mtDNA regions, most of which had not been previously described. A total of seven composite haplotypes were detected. Nucleotide diversity was relatively low for all coding genes but slightly higher for the control region. Microsatellite analysis revealed a relatively high value of major allele frequency, lower values of observed and expected heterozygosity, as well as a moderately reduced number of alleles and genotypes in three of the five observed loci. Although with a clear trend of decline, the level of genetic diversity is still sufficient to ensure the subsistence of the population if the stressors are removed. Otherwise, the loss of heterozygosity will continue, possibly to the point of a complete eradication of the Dalmatian barbelgudgeon from the Busko Blato reservoir.