The hyaluronidase enzyme has been used in many such fields of medicine as ophthalmology, orthopaedia, internal medicine, gynecology, surgery, oncology and dermatology. In this study, the hyaluronidase enzyme was purified and characterized for the first time from Egyptian bee venom homogeneously using DEAE-cellulose and Sephacryl S-300 columns. Bee venom hyaluronidase specific activity was 411.7 units/mg protein with 49.9% yield and 3.23-fold purification. The molecular weight of the purified bee venom hyaluronidase native form was 37 kDa. The purified enzyme was found homogeneous on native PAGE and SDS-PAGE, with two congruent subunits of 18.4 kDa and isoelectric point (pI) of 8.6–8.8. The enzyme was found to be stable over a wide range of temperature (20–60°C) and pH (4.5–6.5), and its optimum activity at 37°C, pH 5.4 and 0.15 M NaCl. Km for bee venom hyaluronidase was 0.029 mg/ml hyaluronic acid and its activity was elevated in presence of MgCl2 and ZnCl2 and lowered in presence of FeCl2. Heparin inhibited the hyaluronidase enzyme noncompetitively with a Ki value of 2.9 units heparin and one binding site on the enzyme molecule.
In bumble bee colonies, pollen is the only protein source for larval feeding and its shortage causes a distress in larval development. Adult bumble bees need pollen for the development of glands and the reproductive system. In bumble bee rearing, honey bee collected pollen is used as the main protein source, either as fresh-frozen or dried pellets, and pollen provisioning is the most problematic and expensive aspect of mass rearing. In honey bee breeding, pollen substitutes are used during the period of food shortage or to stimulate colony strength. We tested different protein diets (five commercial pollen substitutes and two natural protein sources) for the maintenance of bumble bee colonies in captivity. We further mixed Feedbee®, one of the substitutes that gave the best results, with different amounts of pollen to evaluate the optimal amount needed for the whole colony development. Although none of the pure protein diets alone were adequate, diets with a 1 to 1 and 1 to 3 ratio of Feedbee to pollen were both suitable for colony development and queen production. The colony consumed between 2 and 4 g per day of the Feedbee mixed diets, corresponding to a protein consumption of 0.75–0.85 g day−1. Nevertheless, the consumption rate of the pure pollen showed that a mean amount of protein between 0.4 and 0.5 g day−1 was enough to allow colony development indicating the suitability of Feedbee mixed diets.
The aim of the study is to present all distribution sites of Bombus semenoviellus in Poland reported between 1988 and 2019. The increase of species dispersion was concluded, especially in the central part of the country, as well as the occurrence in the high Karkonosze and Tatry mountains. This is likely the effect of progressing climate change, as well as the increase in climate continentalism in Europe. Differences between summer and winter temperatures favour the spread of this Siberian bumblebee species throughout the continent. Many specimens were found on plants originating from its central and west Asia. Those plants also occur in eastern and central Europe which could have helped this bumblebee spread west throughout Europe. B. semenoviellus will increase its range towards Western Europe. Based on observations so far, it is not possible to determine its invasive features and increased competitiveness for food and nesting places in relation to other bumblebee species.
Apis mellifera L. is an essential pollinator that is currently being affected by several stressors that disturb their ecological function and produce colony losses. Colonies are being seriously affected by the ectoparasitic mite Varroa destructor. The relationship between stressors and bee symbionts is being studied in order to enhance bee health. The goal of this study was to evaluate the effect of cell-free supernatants (CFSs) produced by Lactobacillus johnsonii AJ5, Enterococcus faecium SM21 and Bacillus subtilis subsp. subtilis Mori2 on A. mellifera nutritional parameters and their toxicity against V. destructor. Toxicity and survival bioassays were conducted on adult bees with different concentrations of CFSs. Nutritional parameters such as soluble proteins and fat bodies in abdomens were measured. Varroa destructor toxicity was analyzed by a contact exposure method and via bee hemolymph. At low concentrations, two of CFSs tends to enhance bee survival. Remarkably fat bodies maintained their levels with all CFS concentrations in the abdomens, and soluble protein increased at a high concentration of two CFSs. Toxicity against V. destructor was observed only via hemolymph, and results were in agreement with the treatment that produced an increase in bee proteins. Finally, CFS produced by L. johnsonii AJ5 could be a promising natural alternative for strengthening bee health.
The three-year study on borage was conducted in Lublin, SE Poland. The aims were to investigate the flowering pattern and abundance, and the attractiveness (in terms of nectar and pollen production) for flower-visiting insects, mainly bees. Insect visitation and the effect of pollinators on fruit set and seed set were assessed as well. Flowering of borage started in the latter half of June and lasted eight weeks. The mean number of flowers · m−2 of the crop was 4570 per season. A borage flower produced on average 4.0 mg of nectar with a mean sugar concentration of 31.5%. The mean total sugar amount secreted in nectar was 1.2 mg. The pollen amount · flower−1 was 1.1 mg. A borage plant can supply insects with 1.1 g of nectar sugars and 1.1 g of pollen. The estimated nectar sugar yield and pollen yield per 1 m2 of the crop were similar, i.e. 5.2 g. Bees accounted for 73.0% of all insect visits to the borage flowers. The presence of insect pollinators increased the fruit set by 43.3% and seed set by 26.8%.
Oil peony (Paeonia spp.) is a new type of woody oil crop in China with a large cultivation area. Inadequate pollination is one of the main reasons for low seed yield. A pollination net room was built at an oil tree peony base, the numbers of honeybees (Apis mellifera L.) and ground bumblebees (Bombus terrestris L.) were artificially increased and the foraging behaviors and daily activities of the two bees on the plants were observed. Four different pollination methods (honeybee pollination, ground bumblebee pollination, natural field pollination and pollination without insects) were applied. The visit interval, single-flower visit time, number of single-flower visits, number of flowers visited per minute and number of stigma contacts were compared. Meanwhile, the effects of honeybee and bumblebee pollination on the oil yield and quality of peony seeds were compared. There were noticeable differences in daily activity between honeybees and ground bumblebees. Significant differences in the single-flower visit time, visit interval and visit frequency were also observed; honeybee and ground bumblebee pollination increased the seed yield of oil tree peony by 78.74% and 31.88%, respectively. Therefore, both honeybees and ground bumblebees are effective pollinators of oil tree peony. These results provide a theoretical basis for further utilization of bee resources for oil tree peony pollination.
Organophosphate insecticides are known to inhibit the activity of enzyme acetylcholinesterase. They affect olfactory learning and memory formation in honeybees. These insecticides cause mushroom body inactivation in honeybees, but their influence on other brain regions involved in olfactory perception and memory is unknown. The goal of this study was to study the effects of organophosphate insecticide diazinon on carnolian honeybee (Apis mellifera carnica) acetylcholinesterase activity in the olfactory brain regions of antennal lobe, mushroom body and lateral procerebrum (lateral horn). The lamina, medulla and lobula of optic lobes were also analyzed. The level of acetylcholinesterase activity was visualized using the histochemical staining method. Densitometric analysis of histochemical signals indicated that diazinon inhibited acetylcholinesterase activity only in the lip of calyces of mushroom body, but not in other analyzed olfactory regions, antennal lobe and lateral procerebrum. The visual brain system optic lobes were also unaffected. This is in accordance with the literature reporting that mushroom body is the main brain center for olfactory learning and memory formation in honeybees.
Beekeepers need to establish migratory apiaries to benefit from pollen and nectar source plants as in order to increase honey yield. Thus, following the flowering seasons of honey source plants has vital importance when deciding the route of migration. In this study, MODIS imagery was used to generate weekly NDVI data between 1st April to 31st August 2018, when beekeeping activities start and end in the study area. Although MODIS images have high temporal resolution, low spatial resolution (250 meters) makes them insufficient when deciding the crop types and plants. While detecting plants in natural plant areas requires high spatial resolution NDVI, Crop Coverage Registration System (CCRS) parcel-based crop coverage records can enrich the NDVI data without increasing spatial resolution in agricultural lands. Thus, the CCRS data were integrated with NDVI images for migratory beekeeping in agricultural areas as an innovation. To generate both high temporal and spatial resolution, NDVI and CCRS data were integrated together with a beekeeping suitability map to generate the apiary schedule. The results were verified with 176 existing apiary locations and production dates retrieved from field studies which revealed the existence of three seasons in the study area as early and late apiaries (in natural plant areas) and apiaries in agricultural lands. Accuracy analysis showed that 82% of the apiaries intersected with suitable locations and that apiaries in agricultural areas were detected five days earlier than in field studies and obtained more accurately than natural plant apiaries.
Honey bees (Apis mellifera L.) inhabiting trees in forests are not managed by humans or treated for pathogens; therefore, many researchers and beekeepers believe that viral, bacterial, and parasitic diseases may lead to their decline. The aim of the study was to evaluate the prevalence of L. passim and Nosema spp. in feral colonies by real-time PCR. This study was performed on twenty-six samples of honey bees inhabiting tree trunks in north-eastern Poland. One sample consisted of sixty worker bee abdomens collected from hollow trees. Honey bees were sampled only from naturally colonized sites. Amplicons of the three evaluated pathogens were detected in twenty of the twenty-six tested samples. A significant correlation was observed between infection with three pathogens (N. apis, N. ceranae, L. passim) (r = 0.84) compared to infection with only two pathogens (N. apis and N. ceranae) (r = 0.49). N. ceranae was the predominant pathogen, but infections with various severity caused by L. passim were also noted in fourteen of the twenty-six tested samples. In view of the general scarcity of epidemiological data concerning coinfections with Nosema spp. and L. passim in honey bees in tree trunks in other countries, further research is needed to confirm the effect of concurrent pathogenic infections on the decline of bee colonies.
Hemolymph is the “blood” circulating in the entire insect body. Analysis of this fluid gives information about the condition of the insect. The most precise analyses are conducted on insects’ hemolymph suspended in 0.6% physiological saline (NaCl). Most current hemolymph extraction methods are either difficult or do not provide pure material, the contamination of collected insect “blood” can change results. This study aimed to develop a technique for extract hemolymph, both easy and without risk of contaminating derived material. The presented method is a modification of available ways to extract, store and transport hemolymph with the use of a glass capillary. For the development of this technique, adult worker honey bees (Apis mellifera L.) were used. The method required such basic equipment as a glass capillary, sterile tweezer, Eppendorf tube and physiological saline. The collected hemolymph were frozen and hence sent in a glass capillary to another laboratory for analysis.