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Introduction: Brevibacillus laterosporus is a promising microbiological agent that can be used to prevent and control destructive diseases affecting honey bee colonies. In the presentstudy, the short-term effect of the B. laterosporus BGSP11 bee diet on microbiota and mycobiota was investigated. Methods: The honey bee diet was supplemented with spores of B. laterosporus BGSP11 at a concentration of 1×108 CFU/mL in sucrose solution. Metabarcoding analysis of the bee microbial community profile was performed based on 16S RNA (bacteriobiota) and Internally Transcribes Spacer (ITS) region (mycobiota) obtained using MiSeq Illumina sequencing. The QIIME2 v2021.4 pipeline was used to analyze the obtained amplicon data library. Results: The results show that the BGSP11 bee diet slightly altered the bee microbiota and did not lead to potentially harmful changes in the bacterial microbiota. Moreover, it can potentially induce positive changes, mainly reflected in the reduction of opportunistic bacteria. On the other hand, the treatment had a greater effect on mycobiota. However, the changesin the bee mycobiome caused by the treatment cannot be considered a priori as beneficial or harmful,since the interaction between the bee and its mycobiome is not sufficiently studied. The observed positive changes in the bee mycobiome are mainly reflected in the reduction of phytopathogenic fungi that may affect the organoleptic and techno-functional properties of honey. Conclusion: This pilot study suggests that the introduction of BGSP11 in beekeeping practice as a biological agent could be considered due to no harmful effects observed on the microbiota of bees.

In the current study, honey bees’ diet was supplemented with spores of Brevibacillus laterosporus BGSP11 at concentration of 108 CFU/ml in sucrose solution and its short-term effects on their micro- and mycobiota have been analyzed using Illumina MiSeq sequencing. Obtained results indicate that this treatment does not lead to potentially harmful changes in the bacterial microbiome of worker bees, slightly affecting the composition of core microbiota. Moreover, several potentially beneficial changes have been observed. The treatment has led to a significant increase in the abundance of Snodgrassella alvi, and species from Lactobacillus and Bifidobacterium genera which play important roles in protection against several honey bee pathogens. Simultaneously, B. laterosporus enriched diet have led to almost complete eradication of Enterobacteriaceae family, the taxon that contains several putative pathogen species. On the other hand, the treatment affected mycobiota more profoundly, which was expected considering the greater instability compared to microbiota. Although the observed changes in honey bee mycobiome cannot be considered a priori beneficial or harmful, since the interaction between the bee and its mycobiome has not been sufficiently studied, certain beneficial consequences of the treatment have been observed. They are primarily reflected in the reduction of phytopathogenic fungi that can affect the organoleptic and techno-functional characteristics of honey. In addition, before introducing B. laterosporus in beekeeping practice as a biological agent for pathogen control it is necessary to perform more thorough studies of the impact on the honey bee microbiome, immune system, physiology and economic characteristics of honey bee colonies.

Simple Summary The Oriental honey bee Apis cerana, similar to the Western honey bee Apis mellifera, is distributed in different climatic conditions, and each of them is subdivided into more than thirty subspecies and ecotypes. Their sustainability depends on adaptations to the local climate conditions. The VG gene is involved in the development, reproduction, labor division, and caste differentiation of honey bees. We found the nucleotide sequences of the VG gene reflect the adaptation of honey bees to the local climate conditions. The VG gene sequences are acceptable tools to study the sustainability, genetic structure, and adaptation of A. cerana populations and can be applied in conservation genetics of local honey bee subspecies. Apis cerana and Apis mellifera are important honey bee species in Asia. A. cerana populations are distributed from a cold, sharply continental climate in the north to a hot, subtropical climate in the south. Due to the Sacbrood virus, almost all A. cerana populations in Asia have declined significantly in recent decades and have recovered over the past five years. This could lead to a shift in the gene pool of local A. cerana populations that could affect their sustainability and adaptation. It was assumed that adaptation of honey bees could be observed by comparative analysis of the sequences of genes involved in development, labor division, and caste differentiation, such as the gene Vitellogenin VG. The VG gene nucleotide sequences were used to assess the genetic structure and signatures of adaptation of local populations of A. cerana from Korea, Russia, Japan, Nepal, and China. A. mellifera samples from India and Poland were used as the outgroup. The signatures of adaptive selection were found in the local population of A. cerana using VG gene sequence analysis based on Jukes-Cantor genetic distances, cluster analysis, dN/dS ratio evaluation, and Tajima's D neutrality test. Based on analysis of the VG gene sequences, Apis cerana kor

Among them, bacterial and fungal pathogens Paenibacillus larvae, Melissococcus pluton, Ascosphera apis andNosema ceranae play a major impact on honey bees colonies. Thus, developing alternative prophylactic andcurative strategies are urgently needed. The use of probiotic bacteria in honey bee supplemental feeding istherefore promising to treat or prevent diseases. Brevibacillus laterosporus, Gram-positive endospore formingbacilli, is recognised as one of the promising antibacterial and antifungal agents producer.The aim of this study was to examine the short-therm effects of B. laterosporus supplemented diet on workerhoney bee microbiome.Dry spores of B. laterosporus strain BGSP11 have been administrated through a sugar syrup diet to tencolonies and a representative specimen of worker honey bees was taken before the start of the treatmentand immediately after the syrup was consumed. The microbial diversity was assessed before and after thetreatment using Illumina MiSeq sequencing platforms (ID Genomics service, Seattle, WA, USA). 16s rRNAsequencing for bacterial community profiling and fungal Internally Transcribes Spacer for mycological taxaprofiling were used. The next-generation microbiome bioinformatics platform QIIME2 v 2021.4 was used forfiltering and denoising obtained sequences, calculation of diversity metrics and taxonomy assignment. Thefeature classifier was trained using the Greengenes v 13_8 for bacterial taxa and fungal UNITE database v 8.3.The results obtained in this study indicated statisticaly significant alfa diversity between control and experimentalgroup honey bee microbiota composition. The diversity abundance was higher in control comparingto the group treated with B. laterosporus strain BGSP11 spores. There was no significant diference in Bray-Curtis distance among two groups of analysed samples. Regarding to mycological abundance, compositionwas completely different between two groups; control group had Claviceps as predominant genus, while

Simple Summary Honey bee colonies collect pollen from plants as a source of nutrients. Diverse diets comprising pollen from many different plant species are beneficial for honey bee colony health, because they contain a greater diversity of nutrients than monofloral diets of one plant species only. Here, we present the results of the COLOSS "CSI Pollen" study on the diversity of pollen collected by honey bee colonies. In this study, beekeepers acted as citizen scientists sampling and analyzing pollen collected by their own colonies. As a simple measure of diversity, beekeepers determined the number of different colors found in pollen samples that were collected in a coordinated and standardized way. The support of 750 beekeepers allowed the collection of information about almost 18,000 pollen samples from many European countries. We found that the pollen samples consistently comprised approximately six different colors in total, of which four colors were abundant. 'Urban' habitats or 'artificial surfaces' were associated with higher pollen color diversity. This investigation highlights seasonal- and land-use-related differences in the pollen supply for honey bees, which affects beekeeping and pollinator health. Determining pollen colors is a simple, useful technique for beekeepers to estimate pollen diversity. A diverse supply of pollen is an important factor for honey bee health, but information about the pollen diversity available to colonies at the landscape scale is largely missing. In this COLOSS study, beekeeper citizen scientists sampled and analyzed the diversity of pollen collected by honey bee colonies. As a simple measure of diversity, beekeepers determined the number of colors found in pollen samples that were collected in a coordinated and standardized way. Altogether, 750 beekeepers from 28 different regions from 24 countries participated in the two-year study and collected and analyzed almost 18,000 pollen samples. Pollen samples contained approximate

Cilj ovog istraživanja je utvrĎivanje postojanja morfoloških, proizvodnih i genetskih razlika izmeĎu četiri geografski udaljenih selekcionisanih linija medonosne pčele iz Srbije (iz Vršca i Vranja) i Crne Gore (iz Bijelog Polja i Sutomora). TakoĎe ispitivane su proizvodne osobine odabranih linija: broj pčela, površinu legla, meda i polena, produktivnost i higijensko ponašanje pčelinjih društava u periodu trajanja dve pune pčelarske sezone. Ispitivani su i osnovni morfometrijski parametri na telu medonosne pčele i zajedno sa geometrijskom morfometrijom krila pčela determinisane su razlike izmeĎu ispitivanih linija. UtvrĎene su i mere za promere spermateka i broj ovariola ispitivanih matica. Na kraju su karakterisane razlike i sličnosti izmeĎu ispitivanih linija uz pomoć alozimske i DNK analize. Na osnovu analize dobijenih rezultata proizvodnih osobina uz pomoć ANOVE i I-distance izvršeno je rangiranje ispitivanih linija. Najbolje proizvodne rezultate je pokazala linija iz Vršca. U pogledu higijenskog ponašanja najbolje osobine otkrivanja i čišćenja ubijenih larvi je pokazala linija iz Vranja. Pri analiziranju rezultata klasične morfometrije najviše razlika meĎu ispitivaim linijama na svim nivoima statističke značajnosti je utvrĎeno prilikom analiziranja širine bazitarsusa, dužine i širine zadnjeg krila, dok su pri kanonijskoj diskriminantnoj analizi najvećoj interlinijskoj determinaciji doprineli sledeći karakteri: kubitalni indeks, širina zadnjeg krila, dužina femura i širina bazitarzusa. IzmeĎu ispitanih pojedinačnih uzoraka krila je otkrivena niska unutar populacijska varijabilnost. Pri analizi rezultata geometrijske morfometrije je dobijeno mnogo veće razdvajanje i bolji rezultati diskriminantne analize na nivou pčelinjeg društva u poreĎenju sa istom analizom na nivou pojedinačnih uzoraka krila. UporeĎivanjem izmerenih dimenzija za prečnik spermateke se uočava da je najveći prosečni promer bio kod matica iz linije iz Vršca (1,338 mm), a najmanji kod linije iz Vra, The objective of this research is determining the existence of morphological, productive and genetic differences between four geographically distant honeybee selection lines from Serbia (From Vršac and Vranje) and Montenegro (from Bijelo Polje I Sutomore). The productive characteristics of selected lines were also examined: number of bees, brood, honey and pollen surface, productivity and hygienic behavior of honey bee colonies during the period of two full beekeeping seasons. The main morphometric parameters of the honeybee body were also examined together with geometric morphometrics of the bee wing which helped determine the differences between the tested lines. The measures for the spermathecae diameter and number of ovarioles of the tested queens have also been identified. In the end, differences and similarities were characterized between the tested lines using allozyme and DNA analysis. Based on the analysis of the results of productive characteristics, the tested lines were ranked using ANOVA and the I-distance. The best production results were showed by Vrsac lines. In terms of hygienic behavior, the best features of detecting and cleaning of the dead larvae were showed by Vranje lines. When analyzing the results of classical morphometrics, the maximum difference between the examined lines at all levels of statistical significance was found when analyzing basitarsus width, the length and width of the rear wing, while in the canonical discriminant analysis the greatest determination between the lines was contributed by the following characters: cubital index, the width of the rear wing, the length of the femur and basitarsus width. Among the tested individual wing samples, a low variability within the population was detected. Analyzing the results obtained by geometric morphometrics resulted in a much bigger separation and better results of discriminant analysis on bee colony level compared with the same analysis for individual wing samples. Comparing the me