Effect of entomopathogens on Africanized Apis mellifera L. (Hymenoptera: Apidae)

This study aimed to evaluate the effect of commercially used entomopathogens on Africanized Apis mellifera L. (Hymenoptera: Apidae). Four bioassays were performed: 1) pulverized entomopathogens on A. mellifera; 2) entomopathogens sprayed on a smooth surface; 3) entomopathogens sprayed on soy leaves; and 4) entomopathogens mixed with candy paste (sugar syrup). Five treatments were prepared: sterile distilled water (control), distilled water sterilized with Tween^® 80 (0.01%), and the commercial entomopathogens Metarhizium anisopliae E9 (1.0 × 10⁹ conidia mL^?1), Beauveria bassiana PL63 (1.0 × 10⁸ conidia mL^?1) and Bacillus thuringiensis var. kurstaki HD-1 (3.0 × 10⁸ spores mL^?1). Each treatment consisted of five repetitions, with 20 workers per repetition, which were stored in a plastic box and, later, in a biological oxygen demand (B.O.D.) incubator (27 ± 2 °C, RH of 60% ± 10%, 12-h photophase). The mortality of the workers was evaluated from 1 h to 240 h, and the data were analyzed using Bayesian inference. The workers killed by the ingestion of candy paste contaminated with the pathogens (products) were randomly separated and selected for the removal of the midgut. Each midgut was fixed in Bouin's solution and prepared for histology. B. bassiana was verified to reduce the survival of A. mellifera workers in all bioassays. Moreover, M. anisopliae reduced the survival of A. mellifera workers directly sprayed, on a smooth surface and mixed with candy. B. thuringiensis reduced A. mellifera survival on a smooth surface and mixed with candy paste. However, its effects were lower than that observed by B. bassiana. The treatments with the biological products did not induce morphometric alterations in the midgut of A. mellifera.     

Rainbow bee-eaters (Merops ornatus) as a monitoring tool for honeybees (Apis mellifera L.; Hymenoptera: Apidae)

Abstract Regurgitated pellets were collected from underneath roosts of rainbow bee-eaters in suburban Darwin, Australia, and examined for the presence of wings of honeybees. The proportion of pellets containing wings was compared prior to and after placement or removal of honeybee hives in the vicinity of four roosts. On each occasion, the addition or removal of hives was reflected in proportions of pellets containing wings. The results suggest that examination of pellets beneath bee-eater roosts would be a useful technique for monitoring the occurrence of feral honeybees. Potential uses for this technique in eradication of unwanted bees are discussed.     

Effect of Non-Ionizing Electromagnetic Radiation on Behavior of the Honeybee,Apis mellifera L. (Hymenoptera,Apidae)

Entomological Review - We studied the effect of non-ionizing electromagnetic radiation (EMR) of a Wi-Fi router on sensory olfactory excitability, food motivation, and the ability to form a...     

Antennal malformations in light ocelli drones of Apis mellifera (Hymenoptera, Apidae)

Malformed antennae of Apis mellifera light ocelli drones were drawn, dissected and mounted permanently on slides containing Canada balsam, in order to count the olfactory discs present in each segment, in comparison with the number of those structures in normal antennae of their brothers. Some drones presented morphological abnormalities in a single segment of the right or left antenna, but others had two or more malformed segments in a same antenna. Drones with malformations in both antennae were also observed. The 4th and 5th flagellum segments were the most frequently affected. In a low number of cases the frequency of olfactory discs in malformed segments did not differ from that one recorded for normal segments. However, in most cases studied, the antennal malformations brought about a significant reduction in the number of olfactory discs from malformed segments.     

Larval Apis mellifera L. (Hymenoptera: Apidae) mortality after topical application of antibiotics and dusts

Beekeepers apply various dusts to honey bee, Apis mellifera L., colonies to dislodge parasitic mites and control bacterial brood diseases. Anecdotal reports by beekeepers indicate that the antibiotic oxytetracycline (OTC) can be toxic when applied in powdered sugar to cells containing immature bee brood, but it was not known whether the purported toxicity is caused by the antibiotic or the sugar carrier. Additionally, the toxicity of various dusts, proposed for parasitic mite control, is poorly known. In the current studies, we tested OTC and two other antibiotics (tylosin and lincomycin, candidate compounds for use in honey bee colonies) in a powdered sugar carrier for larval toxicity. We also tested for larval toxicity, several dusts that have been proposed for mite control. OTC caused significant brood mortality of approximately 80% at the concentrations used in the hive (200 mg in 20 g sugar). In contrast, tylosin and lincomycin at the 200 mg dose were both similar to untreated controls, and only five times that concentration (1000 mg) caused significant brood mortality of approximately 65%. The addition of dusts, wheat flour, talc, and a commercially available protein supplement, BeePro, resulted in mortality levels between 65 and 80%, similar to that seen with OTC. The common antibiotic carrier, powered confectioners sugar, was nontoxic. The use of 100 unsealed brood cells was demonstrated to be a reliable means of assessing potential adverse affects of dry compounds on larval honey bees. Two new candidate antibiotics for use in honey bee colonies were less toxic to larval bees than the currently labeled antibiotic, OTC.     

Bombus impatiens (Hymenoptera: Apidae): an alternative to Apis mellifera (Hymenoptera: Apidae) for lowbush blueberry pollination.

The pollination effectiveness of the commercially reared bumble bee Bombus impatiens Cresson, was compared in field studies to the honey bee, Apis mellifera L., for lowbush blueberry, Vaccinium angustifolium Ait. A preliminary study indicated that B. impatiens had potential as an alternative pollinator. In a 3-yr study, percentage fruit set, percentage harvested berries, berry weight, and seeds per berry were compared in blueberry fields stocked at 7.5 A. mellifera hives per hectare to 5, 7.5, or 10 B. impatiens colonies per hectare. Percentage of harvested berries (yield) was significantly higher in fields stocked with B. impatiens at 10 colonies per hectare. No other parameters measuring pollinator effectiveness were significantly different at 5, 7.5, or 10 colonies per hectare. Flower handling time was significantly faster for B. impatiens and it more frequently collected blueberry pollen. All parameters of pollinator effectiveness were similar for B. impatiens, A. mellifera, and native wild bees in a follow-up study. Overall, B. impatiens was a suitable alternative to A. mellifera.     

Organization of the antennal lobe in the queen honey bee,Apis mellifera L. (Hymenoptera : Apidae)

Antennal afferent pathway topography was studied in the queen honey bee, Apis mellifera L. (Hymenoptera : Apidae) by staining with cobalt chloride applied directly to the cut antennal axons. Antennal lobe organization in the queen was compared with those in worker and drone bees. The organization is similar in queen and worker bees. For the first time in a female insect the possible existence of a macroglomerulus is shown, which may be involved in the processing of species-specific information. A comparative quantitative neuromorphological study of the glomeruli was performed between young queens (8-day old) and 1-yr-old queens. The mean volume of the glomeruli is 46% greater in the older queens.     

Projection pattern of poreplate sensory neurones in honey bee worker,Apis mellifera L. (Hymenoptera : Apidae)

The projections of olfactory receptor cells of the poreplate sensilla were studied in the worker honey bee, Apis mellifera L. (Hymenoptera : Apidae) by filling single sensilla iontophoretically with cobalt chloride. Successful fillings of individual sensilla lead to staining of one to 22 sensory neurones. All stained receptor cell axons are uniglomerular. Seven fillings of poreplates from the 5th flagellar segment in different animals were compared to analyse the distribution patterns of the receptor cell axons in the antennal lobe. The sensory neurones of individual poreplates project to widely distributed glomeruli in the antennal lobe. The projection patterns of different poreplates are not the same, but may be overlapping.     

意蜂工蜂酸性磷酸酶的纯化及其酶学特性

从意蜂Apis mellifera工蜂体内分离提纯酸性磷酸酶（ACPase, EC3.1.3.2）,并对其性质进行了研究。将工蜂酸性磷酸酶的初提物经分段盐析、DEAE-Sepharose FF离子交换层析及Sephadex G-200 凝胶过滤等纯化步骤,得到经聚丙烯酰胺凝胶电泳为单一蛋白区带的酶液。提纯倍数为77.24,酶液比活力为16.22 U/mg（对硝基苯磷酸二钠作底物）。利用凝胶过滤法测定酶的相对分子质量为135 kD,SDS-PAGE测定酶的亚基相对分子质量为63 .1 kD。酶的等电点为4.46和4.79。非还原/还原（NR/R）单向、双向SDS-PAGE显示酶分子含有链内二硫键。对二级结构圆二色谱分析显示,酶分子中α-螺旋占13.84%,β-折叠占25.68%,无规则卷曲占56.34%。氨基酸组成分析结果表明, 酸性磷酸酶约含有507个氨基酸残基,富含门冬氨酸残基。     

成年蜜蜂脑神经细胞的培养和电生理特征

为了研究杀虫剂等对蜜蜂毒性作用的神经机制,需在体外建立成年蜜蜂脑神经细胞的分离培养和电生理记录技术并研究其正常电生理特征,而对成年蜜蜂脑神经细胞的分离培养和电生理特性的研究报道甚少。我们采用酶解和机械吹打相结合的方法获得了数量较多且活力较好的成年意大利蜜蜂Apis mellifera脑神经细胞,并用全细胞膜片钳技术研究了成年意大利蜜蜂脑神经细胞对电流和电压刺激的反应,获得了成年意蜂脑神经细胞的基本电生理特征以及钠电流和钾电流的特性。全细胞电流钳的记录结果表明,在体外培养条件下,细胞无自发放电发生,注射电流后仅引起细胞单次放电,引起细胞放电的阈电流平均为60.8±63 pA; 细胞动作电位产生的阈电位平均为−27.4±2.3 mV。用全细胞电压钳记录了神经细胞的钠电流和钾电流。钠电流的分离是在电压刺激下通过阻断钾通道和钙通道实现。细胞的内向钠电流在指令电压为−40～−30 mV左右激活,−10 mV达峰值,钠通道的稳态失活电压V_1/2为−58.4 mV; 外向钾电流成份至少包括较小的快速失活钾电流和和较大的缓慢失活钾电流（占总钾电流的80%）,其半激活膜电位V_1/2为3.86 mV,无明显的稳态失活。结果提示缓慢失活钾电流的特征可能是细胞单次放电的机制之一。     

Resistance to Chalkbrood disease in Apis mellifera L. (Hymenoptera: Apidae) colonies with different hygienic behaviour

Chalkbrood disease affects the larvae of honeybees Apis mellifera L. and is caused by the fungus Ascosphaera apis. Infected larvae die when they are stretched in the cap cell and suffer a gradual hardening that ends in a very hard structure (mummie). Several studies have demonstrated that colonies that express an efficient hygienic behaviour (uncapping of cell and subsequent removal of dead brood) exhibit a higher resistance to the disease. However, it remains unclear whether the advantage of hygienic colonies over less hygienic ones lies in the ability to remove mummies or in the early detection of infected larvae and its cannibalization before they harden. To elucidate this aspect, the hygienic behaviour of 24 colonies, which were subsequently provided with pollen cakes containig A. apis, was evaluated. The number of mummies and the number of partially cannibalized and whole larvae in uncapped cells were recorded. The most hygienic colonies controlled the disease better. These colonies also had a higher tendency to uncap cells that contained infected larvae and cannibalize them. The presence of A. apis in partially cannibalized and whole larvae in uncapped cells indicate that the advantage of hygienic colonies over less hygienic ones lies in the early detection of infected larvae death and their quick removal from the cell before they become mummies.     

Fine structure and innervation of the honey stomach (crop) of the honeybee,Apis mellifera L. (Hymenoptera : Apidae)

The present paper describes the ultrastructure and innervation of the honey stomach (crop) of Apis mellifera (Hymenoptera : Apidae). The crop is innervated by 10–20 μm-thick nerves with numerous axons, running longitudinally, and small nerves (1–3 μm, often containing only one distinct axon), running latitudinally. Crop muscles are striated, often branched and have only few multidirectionally arranged myofilaments; they seem to be of the slow-contracting type. Longitudinal and circular muscles produce projections that connect synaptically to small nerves. At these neuromuscular junctions, a high density of synaptic vesicles is found in the axons. A small number of apparently multipolar neurones, stained with Procion Yellow, are irregularly distributed over the crop. The possibility that these neurones work as stretch receptor organs is discussed.     

The effect of essential oils of sweet fennel and pignut on mortality and learning in africanized honeybees (Apis mellifera L.) (Hymenoptera: Apidae)

It was recently discovered that exposure to small concentrations of the essential oils of sweet fennel (Foeniculum vulgare Mill) or pignut [Hyptis suaveolens (L.) Poit] can be used to control aphids. What is not known is whether these oils also influence honeybee behavior. Experiments using both harnessed and free-flying foragers at concentrations used to control aphids showed that bees readily associated the odors with a reward, discriminated between them, and were not repelled. Honeybees, however, would not consume the oils when mixed with sucrose to create an unconditioned stimulus. An experiment in which harnessed bees consumed various concentrations showed that concentrations greater than 50% were detrimental. The experiments reported here provide further evidence supporting the use of conditioning techniques to evaluate the use of essential oils on honey bee behavior.     

Ultrastructure and ontogeny of the mandibular glands of the queen honey bee,Apis mellifera L. (Hymenoptera: Apidae)

Each mandibular gland of the queen bee, Apis mellifera L. (Hymenoptera: Apidae) consists of an axial cavity lined with a thin cuticular intima, secreted by a flat epithelium, and numerous glandular units (type 3), each unit with one duct cell and a large polyploid glandular cell. Mitochondria and endoplasmic reticulum are preponderant organelles. During the ontogeny of the queen bee, the ultrastructure of the glandular cells evolves as her level of pheromonal activity. Variations mainly concern the mitochondrial system. Hence, at the beginning of the imaginal life, the homogeneous population of small mitochondria increases. Towards the 3rd–5th day of the queen's imaginal life, the pheromonal activity increases and the mitochondrial differentiation results in the appearance of giant forms. During the highest activity phase (6 to 18-month-old queens), giant mitochondria, associated with endoplasmic reticulum, invade cytoplasmic areas. In queens aged from 18 to 24 months, the reduction of pheromonal and secretory activities is associated with the reduction of mitochondrial population as well as with the accumulation of lipid droplets and various lytic structures.     

Magnetoreception system in honeybees (Apis mellifera)

Honeybees (Apis mellifera) undergo iron biomineralization, providing the basis for magnetoreception. We showed earlier the presence of superparamagnetic magnetite in iron granules formed in honeybees, and subscribed to the notion that external magnetic fields may cause expansion or contraction of the superparamagnetic particles in an orientation-specific manner, relaying the signal via cytoskeleton (Hsu and Li 1994). In this study, we established a size-density purification procedure, with which quantitative amount of iron granules was obtained from honey bee trophocytes and characterized; the density of iron granules was determined to be 1.25 g/cm(3). While we confirmed the presence of superparamagnetic magnetite in the iron granules, we observed changes in the size of the magnetic granules in the trophycytes upon applying additional magnetic field to the cells. A concomitant release of calcium ion was observed by confocal microscope. This size fluctuation triggered the increase of intracellular Ca(+2) , which was inhibited by colchicines and latrunculin B, known to be blockers for microtubule and microfilament syntheses, respectively. The associated cytoskeleton may thus relay the magnetosignal, initiating a neural response. A model for the mechanism of magnetoreception in honeybees is proposed, which may be applicable to most, if not all, magnetotactic organisms.     

Ultrastructural and cytochemical aspects of metamorphosis in the midgut of Apis mellifera L. (Hymenoptera: Apidae: Apinae)

The midgut of Apis mellifera is remodeled during metamorphosis. The epithelium and, to a lesser extent, the muscular sheath degenerate between the end of the last larval instar and the onset of pupation (prepupa). The larval epithelium is shed to the midgut lumen and digested, while a new epithelium is reconstructed from larval regenerative cells. During pupation, some reorganization still occurs, mainly in brown-eyed pupae. In pharate adult, the midgut wall shows the characteristics of adult, although some cells have pycnotic nuclei. The localization of alkaline and acid phosphatases showed that these enzymes were not involved in the reabsorption of the midgut wall.     

Geographical patterns of mitochondrial DNA variation in Apis mellifera iberiensis (Hymenoptera: Apidae)

An extensive survey of mitochondrial haplotypes in honeybee colonies from the Iberian Peninsula has corroborated previous hypotheses about the existence of a joint clinal variation of African (A) and west European (M) evolutionary lineages. It has been found that the Iberian Peninsula is the European region with the highest haplotype diversity (12 haplotypes detected of the M lineage and 10 of the A lineage). The frequency of A haplotypes decreases in a SW-NE trend, while that of M haplotypes increases. These results are discussed in relation to hypotheses about the African origin of Apis mellifera and an early colonization of west Europe during intermediate Pleistocene glaciation events, followed by a regional differentiation. The extant pattern of haplotype frequency and distribution seems to be influenced at a regional scale by adaptation to local climatic conditions and the mobile beekeeping that has become a large-scale practice during the last decades. Other previous anthropogenic influences (Greek, Roman and Arab colonizations) are thought to be of minor importance in present day populations.     

Seasonal variation of collected pollen loads of honeybees (Apis mellifera L. anatoliaca)

Pollen collected by honeybees foraging in the region of Bursa, Turkey was analysed for a whole year. Pollen loads were collected from the hives of Apis mellifera anatoliaca once a week and were classified by colour. Forty‐one taxa were identified from the pollen analyses of the loads and 14 of these had percentages higher than 1%. Only 2.05% of the total pollen could not have been identified. Dominant taxa include; Brassicaceae (11.19%), Helianthus annuus L. (10.84%), Cichorioideae (8.93%) Salix spp. (7.99%), Rosaceae (7.37%), Centaurea spp. (7.56%), Papaver spp. (7.41%), Knautia spp. (6.99%), Fabaceae (6.01%), Asteraceae (5.73%), Xanthium spp. (2.65%), Chrozophora spp. (2.45%), Plantago spp. (1.56%) and Acer spp. (1.54%) representing 88.23% of the total. Distinct variations in plant usage are seen through the year with initial use of Rosaceae, Salix, and to a lesser extent Brassicaeae. As these groups finish flowering the bees move onto Helianthus annuus, Centaurea through the summer followed by Asteraceae in the late summer and Fabaceae in the autumn. There is a strong reliance on crop species for pollen forage but a number of indigenous species are also seen within the samples. The most productive period for collecting various pollen types, and the ideal period to determine pollen preferences of honey bees was June‐August.