It has been known for many years that in temperate climates the European honey bee, Apis mellifera, exists in the form of two distinct populations within the year, short-living summer bees and long-living winter bees. However, there is only limited knowledge about the basic biochemical markers of winter and summer populations as yet. Nevertheless, the distinction between these two kinds of bees is becoming increasingly important as it can help beekeepers to estimate proportion of long-living bees in hives and therefore in part predict success of overwintering. To identify markers of winter generations, we employed the continuous long-term monitoring of a single honey bee colony for almost two years, which included measurements of physiological and immunological parameters. The results showed that the total concentration of proteins, the level of vitellogenin, and the antibacterial activity of haemolymph are the best three of all followed parameters that are related to honey bee longevity and can therefore be used as its markers.

Instrumental insemination of Apis mellifera L. queens is a widely employed technique used in honeybee breeding that enables the effective control of mating. However, drone semen represents a potential source of honeybee viruses. In this study, 43 semen doses collected from apparently healthy drones, and consequently used in instrumental insemination, were analysed using PCR or RT-PCR to detect the presence of viral genome of 11 honeybee viruses. In 91% of samples, viral infection was detected. The survey revealed genomes of five viruses, namely Deformed wing virus (DWV), Acute bee paralysis virus (ABPV), Black queen cell virus (BQCV), Sacbrood virus (SBV), and A. mellifera filamentous virus (AmFV) in 84%, 19%, 14%, 2%, and 67% of samples, respectively. Single infection (30% of samples) as well as multiple infection (61% of samples) of two, three or four pathogens were also evaluated. To the best of our knowledge, this is the first study describing the presence of the BQCV and SBV genome sequence in drone ejaculate. Phylogenetic analysis of BQCV partial helicase gene sequence revealed the high similarity of nucleotide sequence of described Czech strains, which varied from 91.4% to 99.6%. The findings of our study indicate the possibility of venereal transmission of BQCV and SBV.

In the temperate climates of middle Europe and North America, two distinct honeybee (Apis mellifera) populations are found in colonies: short-living summer bees emerge in spring and survive until summer, whereas long-living winter bees emerge in late August and overwinter. Besides the difference in their life spans, each of these populations fulfills a different role in the colonies and individual bees have distinct physiological and immunological adaptations depending on their roles. For instance, winter worker bees have higher vitellogenin levels and larger reserves of nutrients in the fat body than summer bees. The differences between the immune systems of both populations are well described at the constitutive level; however, our knowledge of its inducibility is still very limited. In this study, we focus on the response of 10-day-old honeybee workers to immune challenges triggered in vivo by injecting heat-killed bacteria, with particular focus on honeybees that emerge and live under hive conditions. Responses to bacterial injections differed between summer and winter bees. The latter induced more intense response, including higher expression of antimicrobial genes and antimicrobial activity, as well as a significant decrease in vitellogenin gene expression and its concentration in the hemolymph. The intense immune response observed in winter honeybees may contribute to our understanding of the relationships between colony fitness and infection with pathogens, as well as its association with successful overwintering.

This work evaluates the in vitro inhibitory activity of 70 essential oils (EOs) in the vapor phase for the control of Chalkbrood disease caused by Ascosphaera apis Maassen ex Claussen (Olive et Spiltoir). Two wild strains isolated from infected honey bee colonies together with one standard collection strain were tested by the microatmosphere method. From 70 EOs, 39 exhibited an antifungal effect against A. apis standard and wild strains. The greatest antifungal action was observed for EO vapors from Armoracia rusticana, followed by Thymus vulgaris, Cymbopogon flexosus, Origanum vulgare and Allium sativum. An investigation of chemical composition by GC-MS revealed, that the most active EOs contained allyl isothiocyanate, citral, carvacrol and diallyl sulfides as the main constituents. The chemical composition plays a key role, as activities of different EOs from the same botanical species were different according to their composition.

Geographical and botanical origin of honeys can be characterized on the basis of physico-chemical composition, sensory properties and on the basis of melissopalynological analysis. No comprehensive description of the characteristics of Czech honey has been published so far. This study provides insights that are important for correct classification. The study analysed 317 samples of authentic honey from randomly selected localities. Due to the diversity of the landscape, the typical honey of the region is blend honey with a predominance of blossom honey. According to the pollen profile and electric conductivity, the honeys were sorted into the following: Brassica honey (BH), Floral honey (FH), Fruit tree honey (PH), Honeydew (HD), Lime tree honey (LH), Robinia pseudoacacia honey (RH), and Trifolium honey (TH). Physico-chemical properties, including higher carbohydrates, were determined for the honeys and their pollen profiles were examined. The physico-chemical properties and pollen profile are partially in compliance with the description of European monofloral honeys, except for RH and TH. Although they had the highest proportion of acacia pollen, amounting to >10% of all the Czech honeys, these RH honeys differ from the European standard, so they cannot be considered acacia honey. Further, PH honeys and FH polyfloral honeys were described. Most honeys contained a significant proportion of rapeseed pollen, which is one of the common agricultural crops grown in the Czech Republic. All the analysed honeys met the parameters defined by the legislation. Due to direct on-site sampling, honeys were characterized by a low 5-(hydroxymethyl)furfural (HMF) content (3.0 mg/kg) and high diastase activity (24.4 DN). Honeydew honeys had the highest proportion of higher carbohydrates, primarily of Melezitose (4.8 g/100 g) and Trehalose (1.3 g/100 g). The presence of higher carbohydrates was also confirmed in LH for Maltose (4.6 g/100 g) and Turanose (2.4 g/100 g).

Melissococcus plutonius is the causative agent of European foulbrood (EFB), an important bacterial disease of honey bee larvae, and various methods have been developed for the detection of this disease. Adult bees are not affected but spread M. plutonius among beehives and serve as a diagnostic tool to detect EFB. Melissococcus plutonius detection based on conventional polymerase chain reaction (PCR) (16S DNA), qPCR (sodA), and barcode sequencing of the 16S RNA V4 region in worker bees from colonies with and without clinical symptoms were compared. We validated these detection tools in terms of the presence/absence of clinical signs of the disease. The comparison of the PCR- and qPCR-based methods showed their usability for confirmation of the disease in both colonies with and without clinical symptoms. Our results revealed that qPCR was more suitable for the confirmation of clinical EFB than conventional PCR and that conventional PCR was better for general screening, including the screening of asymptomatic colonies, than qPCR. Redundancy analyses (tbRDAs) of the microbiome composition showed that the detection limit-based qPCR of M. plutonius revealed a greater variability in the microbiome profiles of worker bees compared with that explained by clinical signs of the disease and PCR detection as factors. The presence of “secondary invaders” (Paenibacillus alvei and Enterococcus) was positively correlated with an increase in the profile of M. plutonius in the worker bee microbiome, whereas Apibacter adventoris and Bartonella apis were negatively correlated. Both types of correlations were found among Lactobacilli.