Multiple virus infections in the honey bee and genome divergence of honey bee viruses

Using uniplex RT-PCR we screened honey bee colonies for the presence of several bee viruses, including black queen cell virus (BQCV), deformed wing virus (DWV), Kashmir bee virus (KBV), and sacbrood virus (SBV), and described the detection of mixed virus infections in bees from these colonies. We report for the first time that individual bees can harbor four viruses simultaneously. We also developed a multiplex RT-PCR assay for the simultaneous detection of multiple bee viruses. The feasibility and specificity of the multiplex RT-PCR assay suggests that this assay is an effective tool for simultaneous examination of mixed virus infections in bee colonies and would be useful for the diagnosis and surveillance of honey bee viral diseases in the field and laboratory. Phylogenetic analysis of putative helicase and RNA-dependent RNA polymerase (RdRp) encoded by viruses reveal that DWV and SBV fall into a same clade, whereas KBV and BQCV belong to a distinct lineage with other picorna-like viruses that infect plants, insects and vertebrates. Results from field surveys of these viruses indicate that mixed infections of BQCV, DWV, KBV, and SBV in the honey bee probably arise due to broad geographic distribution of viruses.     

Microbiology of ripening honey

Two main groups of bacteria, classified as Gluconobacter and Lactobacillus, are present in ripening honey. A third bacterial group, classified as Zymomonas, and several types of yeast are occasionally isolated. Both in natural honey and in synthetic syrup the bacterial population decreases in the course of the ripening process. Lactobacillus and Gluconobacter disappear after minimum moisture (about 18%) is reached, but the former does so sooner than the latter. The presence of these bacteria in different parts of the bee has been also investigated.     

Antibacterial components of honey

The antibacterial activity of honey has been known since the 19th century. Recently, the potent activity of honey against antibiotic-resistant bacteria has further increased the interest for application of honey, but incomplete knowledge of the antibacterial activity is a major obstacle for clinical applicability. The high sugar concentration, hydrogen peroxide, and the low pH are well-known antibacterial factors in honey and more recently, methylglyoxal and the antimicrobial peptide bee defensin-1 were identified as important antibacterial compounds in honey. The antibacterial activity of honey is highly complex due to the involvement of multiple compounds and due to the large variation in the concentrations of these compounds among honeys. The current review will elaborate on the antibacterial compounds in honey. We discuss the activity of the individual compounds, their contribution to the complex antibacterial activity of honey, a novel approach to identify additional honey antibacterial compounds, and the implications of the novel developments for standardization of honey for medical applications.     

Nectar secretion dynamics and honey production potentials of some major honey plants in Saudi Arabia

The contribution of a bee plant species to honey production depends on the plant’s nectar secretion quality and quantity, which is mainly governed by biotic and abiotic factors. The aim of the current study, was to investigate the nectar secretion dynamics and honey production potential of 14 major bee plant species of the target area. We examined the quantity and dynamics of nectar sugar per flower five times a day using a nectar sugar washing technique and direct measuring of nectar with calibrated capillary tubes. The average nectar sugar amount of the species varied from 0.41 mg/flower to 7.7 mg/flower (P < 0.0001). The honey sugar per flower was used to extrapolate the honey production potential per plant and per hectare of land. Accordingly the honey production potential of the species observed to vary from 14 kg/hectare in Otostegia fruticosa to 829 kg/hectare in Ziziphus spina-christi. The nectar secretion dynamics of the species generally showed an increasing trend early in the morning, peaking toward midday, followed by a decline but different species observed to have different peak nectar secretion times. Generally, the tree species secreted more nectar sugar/flower than the herbs. The nectar secretion amount of the species was positively correlated with the ambient temperature, indicating the adaptation of the species to hot climatic conditions. However, different species were observed to have a different optimum temperature for peak nectar secretion. Despite the limited rainfall and high temperature of the area, many plants were found to have good potential for honey production. The monetary value of honey per hectare of the studied honeybee plant species can be of equal or greater than the per-hectare monetary value of some cultivated crops that require numerous inputs. In addition, the information generated is believed to be useful in apiary site selection and to estimate the honey bee colony carrying capacity of an area.     

Making artificial honey using yeast cells from salivary glands of honey bees

The salivary glands of a honey bee, Apis cerana and the yeast cells isolated from these glands were studied for their effects on sucrose solution. This solution exhibited lowered pH and increased levels of fructose and total amino acids as the time of incubation proceeded. The solution thus made was similar to the natural honey.     

Purification and characterization of honey bee vitellogenin

A protocol has been developed for the purification of vitellogenin from the honey bee, Apis mellifera. Purification allows for the first characterization of a vitellogenin from the large order Hymenoptera. Hymenopteran vitellogenins are unusual among insect vitellogenins in that they contain only one type of apoprotein. The honey bee vitellogenin was isolated from hemolymph of honey bee queens by a combination of density gradient ultracentrifugation, ion-exchange chromatography, and affinity chromatography. The native vitellogenin particle is a very high density glycolipoprotein containing approximately 91% protein, 7% lipid, and 2% carbohydrate. Phospholipid and diacylglycerol are the major lipid components. The equilibrium density (1.28 g/ml) is the same as that for Manduca sexta vitellogenin, which contains a much higher proportion of lipid. The covalently bound carbohydrate moiety of the particle is high in mannose. The amino acid composition of vitellogenin is similar to those of vitellogenins from other insect species. The N-terminal amino acid sequence of the apoprotein was determined, the first such sequence for any insect vitellogenin. When analyzed by sodium dodecyl sulfate (SDS)-gel electrophoresis, A. mellifera vitellogenin resolved into a single band with an apparent Mr of 180,000. Gel filtration under reducing and native conditions yielded estimated Mr values of about 300,000.     

Winter survival of individual honey bees and honey bee colonies depends on level of Varroa destructor infestation

Background

Recent elevated winter loss of honey bee colonies is a major concern. The presence of the mite Varroa destructor in colonies places an important pressure on bee health. V. destructor shortens the lifespan of individual bees, while long lifespan during winter is a primary requirement to survive until the next spring. We investigated in two subsequent years the effects of different levels of V. destructor infestation during the transition from short-lived summer bees to long-lived winter bees on the lifespan of individual bees and the survival of bee colonies during winter. Colonies treated earlier in the season to reduce V. destructor infestation during the development of winter bees were expected to have longer bee lifespan and higher colony survival after winter.

Methodology/Principal Findings

Mite infestation was reduced using acaricide treatments during different months (July, August, September, or not treated). We found that the number of capped brood cells decreased drastically between August and November, while at the same time, the lifespan of the bees (marked cohorts) increased indicating the transition to winter bees. Low V. destructor infestation levels before and during the transition to winter bees resulted in an increase in lifespan of bees and higher colony survival compared to colonies that were not treated and that had higher infestation levels. A variety of stress-related factors could have contributed to the variation in longevity and winter survival that we found between years.

Conclusions/Significance

This study contributes to theory about the multiple causes for the recent elevated colony losses in honey bees. Our study shows the correlation between long lifespan of winter bees and colony loss in spring. Moreover, we show that colonies treated earlier in the season had reduced V. destructor infestation during the development of winter bees resulting in longer bee lifespan and higher colony survival after winter.     

The antibacterial activities of honey

Honey is a powerful antimicrobial agent with a wide range of effects. Various components contribute to the antibacterial efficacy of honey: the sugar content; polyphenol compounds; hydrogen peroxide; 1,2-dicarbonyl compounds; and bee defensin-1. All of these elements are present at different concentrations depending on the source of nectar, bee type, and storage. These components work synergistically, allowing honey to be potent against a variety of microorganisms including multidrug resistant bacteria and modulate their resistance to antimicrobial agents. The effectiveness and potency of honey against microorganisms depends on the type of honey produced, which is contingent on its botanical origin, the health of the bee, its origin, and processing method. The application of antibiotics with honey yielded better antimicrobial potential and synergistic effects were noted against biofilms. In medicine, honey has been used in the treatment of surface wounds, burns, and inflammation, and has a synergistic effect when applied with antibiotics. Tissue repair is enhanced by the low pH of honey (3.5–4): causing a reduction in protease activity on the wound site, elevating oxygen release from hemoglobin and stimulating fibroblast and macrophage activity. Furthermore, H₂O₂ has antiseptic effects, and it disinfects the wound site and stimulates production of vascular endothelial growth factor. The use of honey will clean wounds or burn areas from free radicals and reduces scarring and contractures. The anti-inflammatory and antibacterial potential of honey will keep the injured area moist and as such prevents it from deterioration and fibrosis. Honey can promote fast healing and reduce scarring and is very convenient for plastic surgery. Skin maceration is protected by honey due to its high osmolarity and because it keeps the injury moist. In non-infected areas, honey still reduced pain and inflammation. In general, the use of honey in medical settings has reduced economic loss and provided proven economic benefits by lowering direct costs in comparison to conventional treatments and by using less antibiotics, faster healing and less hospitalization stay. This review is intended to provide an overview of the antibacterial activities of honey and its applications.     

蜜蜂肠道微生物研究进展

蜜蜂是全球范围内重要的授粉昆虫,具有很高的经济、生态效益。但是蜜蜂一生中可能会遇到许多挑战。肠道微生物与蜜蜂的健康息息相关,不仅在营养物质的消化吸收方面有重要作用,而且还能抵抗某些病原菌的侵袭以及增强机体免疫力。近年出现的蜜蜂大量死亡现象可能与肠道微生物失调有关。生物以及非生物的因素都可能密切影响着蜜蜂肠道微生物的存活,导致宿主生理功能紊乱甚至死亡。本文综述了蜜蜂的肠道核心微生物组成、分布、变化、影响因素以及主要作用和研究方法,为肠道微生物的研究和利用提供参考。     

Do competing honey bees matter? Dynamics and abundance of native bees before and after honey bee invasion

To provide replicate samples of local bee populations in a nature preserve, light traps operated continuously on Barro Colorado Island (BCI), Panama, collected bees for 17 years, including 10 years following invasion by African Apis mellifera. Honey bees appeared in light traps as the first swarms colonized the Panama Canal area. Their numbers followed seasonal trends shown in inde-pendent studies, thus indicating bee abundance and activity in a large area. No measurable population-level impact of competition between this invading honey bee and native bees, despite many demonstrations of resource competition at flower patch and colony levels, changed annual abundances of all 15 native bee species. Native bee abundance did not decrease, nor did native bees show substantial reciprocal yearly change with honey bee abundance. One strong negative correlation of bee catches with an extremely rainy year was found. However, multiple regression using rainfall and honey bee abundance as the independent variables showed that neither was responsible for bee population change over 17 years. Nearly half the native species declined during a year that displayed peak honey bee number. That competition from honey bees on an island the size of BCI was necessarily reduced below impact levels expected on the mainland is discussed using a model of resource and consumer density, foraging range, and island size.     

Gluconobacters from honey bees

Fifty-sixGluconobacter strains and oneAcetobacter strain were isolated from honey bees and their environment in three different regions in Belgium and identified phenotypically. Polyacrylamide gel electrophoresis of the soluble cell proteins showed that two different types exist within theGluconobacter isolates: strains from type A were found in samples of the three regions, whereas strains from type B were only isolated in two of the three regions. Both types could occur in bees from the same region, from several hives of one bee keeper and from one hive. Strains from type A were almost identical with collection strainG. oxydans subsp.suboxydans NCIB 9018, whereas strains from type B consituted a new protein electrophoretic type within the genusGluconobacter. AlthoughGluconobacter is apparently associated with honey bees, it is not known whether it is important or required for the bees or any hive product.     

Epidemiology of honey bee parasites

The epidemiology of honey bee parasites has been somewhat neglected, but Lynn Royce and Philippe Rossignol describe their unique characteristics. Indeed, it appears that a parasite of social insects has in essence to adapt to two hosts: first, the individual worker within a colony, the numbers of which grow linearly and second, to the colony itself, the actual reproductive 'organism'. Transmission also has vertical and horizontal components. Analysis of tracheal mite populations in particular suggests that intracolony parasite levels are regulated by the swarming behavior of their hosts. Ironically, current and highly productive methods of honey bee management with movable hives curb swarming and may contribute to increasing the spread and the impact of some parasites. This insight may result in changing management practices to reduce the detrimental effects of bee parasites.     

Flight of the honey bee

Summary Using manometric and gas analytical methods oxygen consumption , carbon dioxide production , respiratory quotientRQ, (Fig. 1A-C) and thorax surface temperature difference T _ts (Fig. 3) were determined in single bees. The animals were either sitting in respiratory chambers or were suspended by the scutum, in which case they were resting, walking (turning a small polystyrene ball) or flying in a closed miniature wind tunnel.During resting (sitting in Warburg vessels) at an ambient temperatureT _a=10°C,RQ was 1.01±0.2 (n=905) with variations due to method (Fig. 1D, E).RQ values during walking were determined in single cases. In no case were they significantly different from 1.00. After the first 10 min of flight meanRQ was 1.00±0.04. It was significantly smaller than 1.00 (RQ=0.97) only during the last 5% of long-time flights (mean flight duration 58.8±28.8 min). With the exception of near-exhaustion conditions no signs of fuels other than carbohydrates were found.Metabolic rateP _m was 19.71±21.38 mW g^–1 during resting at 20°CT _a30°C indicating that many resting bees actively thermoregulate at higherT _a. After excluding bees which were actively thermoregulating, by an approximationP _m was 5.65±2.44 mW g^–1 at 20°CT _a30°C. True resting metabolic rate for sitting bees atT _a=10°C was 1.31±0.53 mW g^–1 (Fig. 2A, B).A significant negative correlation was found between relative (specific) oxygen consumption rel and body massM _b at 85 mgM _b150 mg.At 0°CT _ts16.5°C a significant (-0.01) positive correlation was found between and T _ts in single resting bees: T _Ts+0.099, or betweenP _m and T _ts:P _m=1.343 T _ts+0.581 (Fig. 3D) in ml h^–1,P _m in mW,T in °C).During walking (duration 13.15±5.71 min,n=13) at 12.5°CT _a21°C a stable T _ts of 11.41±3.37°C, corresponding to 167 mW g^–1, was reached for 80 to 90% of the walking time (Fig. 4B).During wind tunnel flights of tethered animals the minimal metabolic power measured in exhaustion experiments was 240 mW g^–1. Calculation of factors of increase inP _m is of limited value in poikilotherms, in which true resting conditions are not exactly defined.