Honey bee colony losses: Why are honey bees disappearing?

The Western honey bee (Apis mellifera L., Hymenoptera: Apidae) is a species of crucial economic, agricultural and environmental importance.In the last ten years, some regions of the world have suffered from a significant reduction of honey bee colonies. In fact, honey bee losses are not an unusual phenomenon, but in many countries worldwide there has been a notable decrease in honey bee families. The cases in the USA, in many European countries, and in the Middle East have received considerable attention, mostly due to the absence of an easily identifiable cause.It has been difficult to determine the main factors leading to colony losses because of honey bees’ diverse social behavior. Moreover, in their daily routine, they make contact with many agents of the environment and are exposed to a plethora of human activities and their consequences. Nevertheless, a number of different factors are considered to be contributing to honey bee losses, and recent investigations have established some of the most important ones, in particular, pests and diseases, bee management, including bee keeping practices and breeding, the change in climatic conditions, agricultural practices, and the use of pesticides. The global picture highlights the ectoparasitic mite Varroa destructor as a major factor in colony loss. Last but not least, microsporidian parasites, mainly Nosema ceranae, also contribute to the problem.Thus, it is obvious that many factors are involved in honey bee colony losses globally. Increased monitoring and scientific research should throw new light on the factors involved in recent honey bee colony losses.This review focuses on the main factors which have been found to have an impact on the increase in honey bee colony losses

Methods for Genotyping of the Honey Bee (Apis mellifera L.: Hymenoptera: Apidae) in Bulgaria

Honey bees are insects of great biological, ecological, and economic importance. They are the subject of a variety of scientific studies. As social insects, they are a preferred and widely used model for clarification of the evolution of social behavior. Because of the haplodiploidy as a mechanism for determining gender, differentiation in the functions of individuals in the bee family, and for its economic importance, Apis mellifera is a significant subject of research in the fields of ontogeny, population genetics, and selective breeding. The biological significance of bees is rooted in the fact that they are main pollinators in the natural environment. About 80% of the pollination of entomophilous plants is carried out by Apis mellifera. In all crops, active pollination significantly increases their yields. Honey bees are a valuable economic asset due to the ensemble of their products, which include honey, bee pollen, propolis, royal jelly, and bee venom, used by humans for food and treatment. The main objective of this chapter was to describe the basic methods used for genotyping of Apis mellifera in Bulgaria. These techniques have been useful to produce a system of population criteria, and taxonomically important molecular markers are applicable in future activities related to the preservation and selection of the Bulgarian honey bee

Molecular detection and phylogenetic assessment of six honeybee viruses in Apis mellifera L. colonies in Bulgaria

Honey bee colonies suffer from various pathogens, including honey bee viruses. About 24 viruses have been reported so far. However, six of them are considered to cause severe infection which inflicts heavy losses on beekeeping. The aim of this study was to investigate incidence of six honey bee viruses: deformed wing virus (DWV), acute bee paralysis virus (ABPV), chronic bee paralysis virus (CBPV), sacbrood virus (SBV), kashmir bee virus (KBV), and black queen cell virus (BQCV) by a reverse transcription polymerase chain reaction (RT-PCR). A total of 250 adult honey bee samples were obtained from 50 colonies from eight apiaries situated in three different parts of the country (South, North and West Bulgaria). The results showed the highest prevalence of DWV followed by SBV and ABPV, and one case of BQCV. A comparison with homology sequences available in GenBank was performed by phylogenetic analysis, and phylogenetic relationships were discussed in the context of newly described genotypes in the uninvestigated South Eastern region of Europe. In conclusion, the present study has been the first to provide sequencing data and phylogenetics analyses of some honey bee viruses in Bulgaria

Biotic and Abiotic Factors Associated with Colonies Mortalities of Managed Honey Bee (Apis mellifera)

Despite the presence of a large number of pollinators of flowering plants worldwide, the European honey bee, Apis melifera, plays the most important role in the pollination of a number of crops, including all vegetables, non-food crops and oilseed crops, decorative and medical plants, and others. The experience of isolated cases of complete extinction of honey bees in individual regions has shown that this phenomenon leads to a dramatic pollination crisis and reduced ability or even total inability to grow insect-pollinated crops if relying solely on native, naturally occurring pollinators. Current scientific data indicate that the global bee extinction between the Cretaceous and the Paleogene (Cretaceous-Tertiary) occurred, which led to the disappearance of flowers because they could not produce viable fruit and germinate due to lack of pollination by bees or other animals. From the Middle Ages to the present day, there has been evidence that honey bees have always overcome the adverse factors affecting them throughout the ages, after which their population has fully recovered. This fact must be treated with great care given the emergence of a new, widespread stress factor in the second half of the 20th century—intoxication of beehives with antibiotics and acaricides, and treatment of crops with pesticides. Along with acute and chronic intoxication of bees and bee products, there are other new major stressors of global importance reducing the number of bee colonies: widespread prevalence of pathogenic organisms and pest beetles, climate change and adverse climatic conditions, landscape changes and limitation of natural habitats, intensification of agricultural production, inadequate nutrition, and introduction of invasive species. This report summarizes the impact of individual negative factors on the health and behavior of bees to limit the combined effects of the above stressors

Sequence analysis of the Hex A gene in Jacob sheep from Bulgaria

Background and Aim: Jacob sheep are a rare ancient breed of sheep believed to have originated from the Mediterranean area but which are now kept throughout the world. These sheep have recently attracted medical interest due to the observation of a genetic disorder in the breed that can be used as an animal model of Tay–Sachs disease (TSD). This study aims to detect mutations in the Hexosaminidase A gene in Jacob sheep based on sequence analysis of the 284-bp fragment situated between exon 11 and intron 11 of the gene, a target sequence for site-specific mutation. This is the first study that has investigated Jacob sheep in Bulgaria for gene-specific mutations. Materials and Methods: A total of 20 blood samples were collected from Jacob sheep from the Rhodope Mountains. DNA was isolated from these samples, and a specific 284-bp fragment was amplified. The amplified products were purified using a polymerase chain reaction purification kit and sequenced in both directions. Results: Target sequences were successfully amplified from all 20 investigated sheep. Sequence analysis did not show the homozygous, recessive, missense (G-to-C transition) mutation at nucleotide position 1330 (G1330→C) in exon 11, demonstrating that all of these sheep were a normal genotype (wild-type). Conclusion: Jacob sheep are considered a potentially useful animal model in advancing the understanding of pathogenesis and developing potential therapies for orphan diseases, such as those characterized by mutant GM2 gangliosides. The clinical and biochemical features of the Jacob sheep model of TSD represent well the human classical late-infantile form of this disorder, indicating that the model can serve as a possible new research tool for further study of the pathogenesis and treatment of TSD

Molecular Detection of Nosema spp. in Honey in Bulgaria

Environmental DNA (eDNA) analysis is related to screening genetic material of various organisms in environmental samples. Honey represents a natural source of exogenous DNA, which allows for the detection of different honey bee pathogens and parasites. In the present study, we extracted DNA from 20 honey samples from different regions in Bulgaria and tested for the presence of DNA of the ectoparasitic mite Varroa destructor, as well as Nosema apis and Nosema ceranae. Only Nosema ceranae was detected, showing up in 30% of all samples, which confirms the widespread prevalence of this pathogen. All positive samples were found in plain regions of the country, while this pathogen was not detected in mountainous parts. None of the samples gave positive amplifications for the Nosema apis and Varroa mite. The obtained results from this study confirm previous observations that eDNA contained in honey is a potent source for effective biomonitoring of actual diseases in the honey bee

Evidence for Early European Neolithic Dog Dispersal: New Data on Southeastern European Subfossil Dogs from the Prehistoric and Antiquity Ages

The history of dog domestication is still under debate, but it is doubtless the process of an ancient partnership between dogs (Canis familiaris) and humans. Although data on ancient DNA for dog diversity are still incomplete, it is clear that several regional dog populations had formed in Eurasia up to the Holocene. During the Neolithic Revolution and the transition from hunter-gatherer to farmer societies, followed by civilization changes in the Antiquity period, the dog population structure also changed. This process was due to replacement with newly formed dog populations. In this study, we present for the first time mitochondrial data of ancient dog remains from the Early Neolithic (8000 years before present (BP)) to Late Antiquity (up to 3th century AD) from southeastern Europe (the Balkans). A total of 16 samples were analyzed, using the mitochondrial D-loop region (HVR1). The results show the presence of A (70%) and B (25%) clades throughout the Early and Late Neolithic Period. In order to clarify the position of our results within the ancient dog population in Eneolithic Eurasia, we performed phylogenetic analysis with the available genetic data sets. This data showed a similarity of the ancient Bulgarian dogs to Italian (A, B, and C clades) and Iberian (clades A and C) dogs’ populations. A clear border can be seen between southern European genetic dog structure, on the one hand, and on the other hand, central-western (clade C), eastern (clade D) and northern Europe (clades A and C). This corresponds to genetic data for European humans during the same period, without admixture between dog populations. Also, our data have shown the presence of clade B in ancient Eurasia. This is not unexpected, as the B haplogroup is widely distributed in extant Balkan dogs and wolves. The presence of this clade both in dogs and in wolves on the Balkans may be explained with hybridization events before the Neolithic period. The spreading of this clade across Europe, together with the A clade, is related to the possible dissemination of newly formed dog breeds from Ancient Greece, Thrace, and the Roman Empire

Genetic Diversity and Structure of the Main Danubian Horse Paternal Genealogical Lineages Based on Microsatellite Genotyping

The Danubian horse, together with the Pleven and the Eastern Bulgarian horse breeds, is one of the modern breeds in Bulgaria. The objective of this study was to compare the genetic structure and genetic diversity of six paternal genealogical lineages of the Danubian horse breed (Zdravko, NONIUS XVII-30, Torpedo, Lider, Kalifa, and Hrabar). In total, 166 individuals from the six genealogical lines were investigated, based on 15 STR markers (short tandem repeats, also known as microsatellites). In total, 184 alleles were found in the six populations, using 15 microsatellite loci. The mean number of alleles, the effective number of alleles, and the polymorphic information content (PIC) values per locus were 12.28, 9.48, and 0.73, respectively. In a comparison of the allelic diversity among sire lineages, the highest genetic diversity (Na) was observed in Lider and Kalifa (14.60 ± 0.21), while the lowest value of this parameter was observed in the Zdravko lineage 4.20 ± 0.35. The largest genetic diversity was found in loci HMS3 and HMS7, with 13 alleles, and the smallest polymorphism was noted for the locus ASB17, with 10 alleles. The level of observed heterozygosity was in the range of 0.65 ± 0.069 for the Zdravko lineage to 0.93 ± 0.01 for the Torpedo lineage. The expected heterozygosity level range was from 0.57 ± 0.048 to 0.91 ± 0.01 for all horse lineages. Structure analysis revealed three main gene pools in the study population. The first pool included the Zdravko lineage; the second had the NONIUS XVII-30, Torpedo, Lider, and Kalifa lineages; and the third defined the Hrabar lineage, which was significantly differentiated from the other genealogical lineages

The first report of the prevalence of Nosema ceranae in Bulgaria

Nosema apis and Nosema ceranae are the two main microsporidian parasites causing nosematosis in the honey bee Apis mellifera. The aim of the present study is to investigate the presence of Nosema apis and Nosema ceranae in the area of Bulgaria. The 16S (SSU) rDNA gene region was chosen for analysis. A duplex PCR assay was performed on 108 honey bee samples from three different parts of the country (South, North and West Bulgaria). The results showed that the samples from the northern part of the country were with the highest prevalence (77.2%) for Nosema ceranae while those from the mountainous parts (the Rodopa Mountains, South Bulgaria) were with the lowest rate (13.9%). Infection with Nosema apis alone and co-infection N. apis/N. ceranae were not detected in any samples. These findings suggest that Nosema ceranae is the dominant species in the Bulgarian honey bee. It is not known when the introduction of Nosema ceranae in Bulgaria has occurred, but as in the rest of the world, this species has become the dominant one in Bulgarian Apis mellifera. In conclusion, this is the first report for molecular detection of Nosema infection of honey bee in Bulgaria. The results showed that N. ceranae is the main Nosema species in Bulgaria