Recipe for a Busy Bee: MicroRNAs in Honey Bee Caste Determination

Social caste determination in the honey bee is assumed to be determined by the dietary status of the young larvae and translated into physiological and epigenetic changes through nutrient-sensing pathways. We have employed Illumina/Solexa sequencing to examine the small RNA content in the bee larval food, and show that worker jelly is enriched in miRNA complexity and abundance relative to royal jelly. The miRNA levels in worker jelly were 7–215 fold higher than in royal jelly, and both jellies showed dynamic changes in miRNA content during the 4^th to 6^th day of larval development. Adding specific miRNAs to royal jelly elicited significant changes in queen larval mRNA expression and morphological characters of the emerging adult queen bee. We propose that miRNAs in the nurse bee secretions constitute an additional element in the regulatory control of caste determination in the honey bee.     

Sterols of organs involved in brood food production and of royal jelly in honey bees

The sterols of the organs (hypopharyngeal and mandibular glands and honey stomachs) involved in worker and queen honey bee brood food production, royal jelly and intact nurse bees were analyzed to obtain information on the selective transfer of specific sterols from one generation to the next. No appreciable increase in the percentage of 24-methylenecholesterol, relative to the total sterols isolated from intact honey bee (Apis mellifera L.), prepupae or adults, was found in the hypopharyngeal or mandibular glands or the honey stomachs from nurse bees reared in colonies fed a chemically-defined diet supplemented with 24-methylenecholesterol. The sterols of these organs contained higher levels of cholesterol than did the sterols of whole body extracts. The other major sterols, sitosterol and isofucosterol, occurred at relative concentrations comparable to whole body extracts. Also, there were higher levels of cholesterol in the sterols from glandular tissues of nurse bees maintained on pollen and sucrose solution than in sterols isolated from intact insects. In a separate study, royal jelly collected over a 6-day period had much higher relative percentages of 24-methylenecholesterol and lower levels of sitosterol and isofucosterol than did the pollen fed to these colonies. The sterols of nurse bees in the latter study had an intermediate concentration of 24-methylenecholesterol. The significance of these findings relative to the unique selective transfer of specific sterols from the diet or from endogenous sterol pools of the nurse bees from generation to generation in the honey bee is discussed.     

Age at Which Larvae Are Orphaned Determines Their Development into Typical or Rebel Workers in the Honeybee (Apis mellifera L.)

In the honeybee, diploid larvae fed with royal jelly develop into reproductive queens, whereas larvae fed with royal jelly for three days only and subsequently with honey and pollen develop into facultatively sterile workers. A recent study showed that worker larvae fed in a queenless colony develop into another female polyphenic form: rebel workers. These rebel workers are more queenlike and have greater reproductive potential than normal workers. However, it was unclear whether larvae orphaned at any time during their feeding period can develop into rebels. To answer this question, the anatomical features of newly emerged workers reared in queenless conditions at different ages during the larval period were evaluated. Our results showed that larvae orphaned during the final four or more days of their feeding life develop into rebel workers with more ovarioles in their ovaries, smaller hypopharyngeal glands, and larger mandibular and Dufour’s glands compared with typical workers with low reproductive potential that were reared with a queen or orphaned at the third to last or a later day of feeding life.     

不同人工代花粉对蜂群群势和生产性能的影响

早春将群势、蜂王年龄和质量基本一致的25群意大利蜜蜂Apis melliferaL.,随机分为5个处理,分别饲喂纯花粉和4种不同的人工代用花粉,观测它们对蜂群群势和生产性能的影响。结果表明,含有添加剂的各代用花粉组与纯花粉组的蜂群的蜂子数量差异均不显著,但均显著高于纯豆粕组(P<0.01);各组产蜜量随着纯花粉所占比例的减少而呈现递减的趋势,但饲喂含有60%花粉+40%豆粕+添加剂1组与纯花粉组没有明显的差异;含有添加剂的各代用花粉组的王台接受率均显著高于纯花粉组(P<0.01);饲喂含有添加剂的各代用花粉组与饲喂纯花粉组相比,王浆产量及蜂王浆中的10-HDA、水分、蛋白质的含量及酸度均没有差异。     

Effects of the mixed biocide Bacillus thuringiensis–abamectin on the development of the parasitoid Microplitis mediator and its host Helicoverpa armigera

The effects of the mixed biocide Bacillus thuringiensis Berliner with abamectin (BtA) on the development of the parasitoid Microplitis mediator (Haliday) (Hymenoptera: Braconidae) and its cotton bollworm host, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), were evaluated in the laboratory. Weight gain in larvae of H. armigera was initially delayed, but larval developmental period increased and pupal weight increased when they were fed on a diet containing BtA. Due to increased longevity of the host larvae, the susceptible period to parasitization of H. armigera by M. mediator increased when the host larvae were reared on diets containing BtA at concentrations of 0.5, 1, 2, and 4 μg g^?1. The longevity of female and male parasitoids significantly decreased when newly emerged wasps were fed a honey solution containing 200 μg ml^?1 BtA in comparison with those fed only a honey solution. Mean longevity was significantly prolonged when parasitoids were fed a honey solution and BtA–honey solution in comparison with those fed BtA–distilled water, distilled water, or nothing. There were no significant differences compared with the control in any biological characteristics for the offspring of female parasitoids fed the honey solutions containing BtA at concentrations of 50, 100, and 200 μg ml^?1; characteristics measured include the egg‐larval period, pupal weight, male and female pupal periods, adult fresh weight, and adult longevity. When female parasitoids parasitized host larvae that had been fed the diet containing BtA, their male and female pupal periods were significantly prolonged compared with the control (without BtA).     

Nephroprotective effect of bee honey and royal jelly against subchronic cisplatin toxicity in rats

Cisplatin is one of the most potent and effective chemotherapeutic agents. However, its antineoplastic use is limited due to its cumulative nephrotoxic side effects. Therefore, the present study was undertaken to examine the nephroprotective potential of dietary bee honey and royal jelly against subchronic cisplatin toxicity in rats. Male Wistar rats were randomly divided into controls, cisplatin-treated, bee honey-pretreated cisplatin-treated and royal jelly-pretreated cisplatin-treated groups. Bee honey and royal jelly were given orally at doses of 20 and 100 mg/kg, respectively. Subchronic toxicity was induced by cisplatin (1 mg/kg bw, ip), twice weekly for 10 weeks. Cisplatin treated animals revealed a significant increase in serum level of renal injury products (urea, creatinine and uric acid). Histopathologically, cisplatin produced pronounced tubulointerstitial injuries, upregulated the fibrogenic factors, α-smooth muscle actin (α-SMA) and transforming growth factor β1(TGF-β1), and downregulated the cell proliferation marker, bromodeoxyuridine (Brdu). Dietary bee honey and royal jelly normalized the elevated serum renal injury product biomarkers, improved the histopathologic changes, reduced the expression of α-SMA and TGF-β1 and increased the expression of Brdu. Therefore, it could be concluded that bee honey, and royal jelly could be used as dietary preventive natural products against subchronic cisplatin-induced renal injury.     

Diet and cell size both affect queen-worker differentiation through DNA methylation in honey bees (Apis mellifera, Apidae)

Background

Young larvae of the honey bee (Apis mellifera) are totipotent; they can become either queens (reproductives) or workers (largely sterile helpers). DNA methylation has been shown to play an important role in this differentiation. In this study, we examine the contributions of diet and cell size to caste differentiation.

Methodology/Principal Findings

We measured the activity and gene expression of one key enzyme involved in methylation, Dnmt3; the rates of methylation in the gene dynactin p62; as well as morphological characteristics of adult bees developed either from larvae fed with worker jelly or royal jelly; and larvae raised in either queen or worker cells. We show that both diet type and cell size contributed to the queen-worker differentiation, and that the two factors affected different methylation sites inside the same gene dynactin p62.

Conclusions/Significance

We confirm previous findings that Dnmt3 plays a critical role in honey bee caste differentiation. Further, we show for the first time that cell size also plays a role in influencing larval development when diet is kept the same.     

Oral Administration of Royal Jelly Restores Tear Secretion Capacity in Rat Blink-Suppressed Dry Eye Model by Modulating Lacrimal Gland Function

Tears are secreted from the lacrimal gland (LG), a dysfunction in which induces dry eye, resulting in ocular discomfort and visual impairment. Honey bee products are used as a nutritional source in daily life and medicine; however, little is known about their effects on dry eye. The aim of the present study was to investigate the effects of honey bee products on tear secretion capacity in dry eye. We selected raw honey, propolis, royal jelly (RJ), pollen, or larva from commercially available honey bee products. Tear secretion capacity was evaluated following the oral administration of each honey bee product in a rat blink-suppressed dry eye model. Changes in tear secretion, LG ATP content, and LG mitochondrial levels were measured. RJ restored the tear secretion capacity and decrease in LG ATP content and mitochondrial levels to the largest extent. Royal jelly can be used as a preventative intervention for dry eye by managing tear secretion capacity in the LG.     

Larval diet influence on oviposition behaviour inSpodoptera littoralis

Females ofSpodoptera littoralis Boisd. (Lepidoptera: Noctuidae) with different feeding experiences during their larval development were tested for their ovipositional response to methanol extracts of larval frass and semisynthetic diets. The effect of the following frass, diet and diet component extracts was tested: (a) frass fromS. littoralis orAgrotis segetum larvae fed on a potato-based diet; (b) frass fromS. littoralis larvae fed on a wheat germ-based diet; (c) potato and wheat germ-based diets; and (d) potatoes and wheat germ. Ovipositing females without prior experience of the potato diet were deterred by extracts of: (1) larval frass from either species fed on potato diet; (2) the potato-based diet; (3) potato. Also females with experience of the potato diet during only a part of their larval development were deterred from oviposition by frass of larvae reared on the potato diet and by the diet itself. However, for females reared on the potato diet for their entire larval development, oviposition was no longer deterred by either of the three extracts listed above. Extracts of: (1) frass from larvae of either species reared on wheat germ diet: (2) the wheat germ diet; or (3) wheat germ did not significantly affect oviposition. Females with ablated antennae were still deterred by frass extracts from larvae fed on potato diet, when they had been reared on the wheat germ diet. In feeding experiments, larvae of larval stage one and of larval stage three-four reared on either of the two diets preferred to feed on the wheat germ diet. However, the preference was significantly stronger for larvae with no prior contact with the potato diet. The effect of larval experience on the loss of oviposition-deterring activity by extracts of larval frass, diets and diet components is discussed in view of induction and selection.     

Ultrastructural analysis on the gland secretion in the extracellular ducts of the hypopharyngeal glands of the honeybee infected by Nosema apis

The hypopharyngeal glands of the honeybee secrete the larval food royal jelly, along with enzymes that are used in the production of honey from nectar. The composition of this secretion mixture varies with the physiological condition of the honeybee, and the final secretion mixture is conveyed through the extracellular ducts to the large excretory duct. The secretion products in the extracellular duct of both healthy and infected honeybees, was studied by transmission electron microscopy and image analysis. These products consisted of a densely packed, faintly visible granular substance. The granules were clearly defined by the computer enhanced image. The secretion products from the Nosema-infected honeybees exhibited the same granular structure, but were much less concentrated, thereby reducing the overall electron density of the secretion. These observations suggest that the composition of the secretion products from the hypopharyngeal glands in the Nosema-infected honeybees, was altered considerably.     

Effects of mixed prey on the development and demographic attributes of a generalist predator,Coccinella septempunctata (Coleoptera: Coccinellidae)

Coccinella septempunctata Linnaeus, 1758 (Coleoptera: Coccinellidae) is an aphidophagous ladybird beetle species with cosmopolitan distribution. However, it may also thrive on arthropods other than aphids, when the latter are not readily available. Certain life history traits and demographic attributes of C. septempunctata were determined using seven different diets with different alternations and proportions of an aphid, Lipaphis erysimi (Kaltenbach,1843) and a mite, Tetranychus urticae Koch, 1836, in the laboratory (at 25±0.5°C, 65±5% RH and a photoperiod of 16 h L:8 h D).The larval duration was longest (12.61±6.17 days) when fed an alternating diet of two days aphid/two days mite. Adult longevity was longest (78.65±1.09 days) when fed aphids only and shortest (3.17±1.25 days) when fed mites only. Life expectancy (e_x) was highest (111 days) when adults were fed only aphids and lowest (11.3 days) when fed only mites. Survival rates of ladybird beetles were positively related to an increasing ratio of aphids in their diet. Their ability to prey on both aphids and mites indicates the effectiveness of C. septempunctata as a biological control agent on plants infested with these two pests.     

蜂王浆主蛋白研究进展

作为社会性昆虫,蜜蜂是研究社会行为和学习记忆的理想模式生物。王浆主蛋白（Major royal jelly protein, MRJP）是蜂王浆中蛋白质的主要成分,该家族一共有9个成员,MRJP1~MRJP9。所有mrjps均以串联排列的形式位于蜜蜂11号染色体上一个大约60 kb的DNA片段上。mrjp的同源体也存在于其他的膜翅目昆虫,均是通过yellow进化而来的。随着不断地进化,MRJPs家族进化出许多重要功能,其中最主要的就是营养功能。本文从MRJPs家族的基因及蛋白质结构、mRNA表达情况、进化和功能等方面进行综述,为今后开展相关研究提供理论支持。     

Worker honey bee ovary development: seasonal variation and the influence of larval and adult nutrition

We examined the effect of larval and adult nutrition on worker honey bee (Apis mellifera L.) ovary development. Workers were fed high or low-pollen diets as larvae, and high or low-protein diets as adults. Workers fed low-protein diets at both life stages had the lowest levels of ovary development, followed by those fed high-protein diets as larvae and low- quality diets as adults, and then those fed diets poor in protein as larvae but high as adults. Workers fed high-protein diets at both life stages had the highest levels of ovary development. The increases in ovary development due to improved dietary protein in the larval and adult life stages were additive. Adult diet also had an effect on body mass. The results demonstrate that both carry-over of larval reserves and nutrients acquired in the adult life stage are important to ovary development in worker honey bees. Carry-over from larval development, however, appears to be less important to adult fecundity than is adult nutrition. Seasonal trends in worker ovary development and mass were examined throughout the brood rearing season. Worker ovary development was lowest in spring, highest in mid-summer, and intermediate in fall.     

Mass rearing of the pink corn borer, Sesamia cretica Led. (Lepidoptera: Noctuidae) larvae, on semi artificial diets

The effect of two different semi-artificial diets (S.A.D1 and S.A.D2) as well as a natural corn diet on the biology and bionomics of the pink stem borer, Sesamia cretica was studied under laboratory conditions. The insect was successfully mass reared for ten successive generations at the conditions of 27 +/- 2 C degrees and 60-80% R.H. In addition, a photoperiod of 0:24 (L:D) for larvae and 12:12 (L:D) in concern to the other stages, respectively. ALong the ten successive generations, there were no significant differences between the larval periods for both the artificial diets. The S.A.D2 induced the shortest (24.0 days) larval period compared with the natural diet (27.5 days) and S.A.D1 (31.5 days). Rearing larvae on the natural diets revealed a pupal period of 11.0 days, while it was 10.0 days and 8.5 days for the artificial diets (S.A.D1) and in (S.A.D2), in respect. Moth longevity (pre-oviposition, oviposition and post-oviposition periods), to a certain extent, was affected by the larval diets. The maximal moth longevity (13.0 days) was observed for larva reared on corn plants (4, 8 and 1 days). On the other hand, the shortest period (10.5 days) of moth longevity was noticed in SAD2 (1, 9 and 0.5 days). S.A.D1 showed 11.0 days moth longevity (3,7 and 1 days). The longest oviposition period (9.0 days) was recorded in the artificial diet S.A.D2, while it was only 7.0 days in the artificial diet S.A.D1 compared with 8.0 days in case of the natural diet. The number of deposited eggs increased with the progress of the 10 successive generations (G1:G10) from 150 to 265 and from 384 eggs to 564 eggs / female for the S.A.D1 and S.A.D2, in sequence. The highest mean number of deposited eggs per female (564) was for the derived females from larvae fed on the artificial diet in G10 of S.A.D2. The rate of the deposited eggs in S.A.D2 was gradually increased (from 9.70 to 61.14% increase) more than those reared on the natural diet throughout the ten generations. The highest significant percent hatchability (92.69%) has been resulted from the artificial diet SAD2. But, it has been decreased to 85.59% in the artificial diet S.A.D1 in comparison to 65.71% in the natural diet. In addition, the significant shortest total generation period was 44.0 days resulted from the artificial diet S.A.D2, while it was 53.5 and 54.0 days for corn plants and the artificial diet S.A.D1, consequently. In short, results indicated that the S.A.D2 could be considered as a suitable artificial diet for a feasible mass rearing of the pink corn borer, Sesamia cretica led. The S.A.D2 showed the shortest larval, pupal and life span for the generation periods. Moreover, it induced the longest oviposition period and the highest mean number of deposited eggs per female and the highest significant percent of eggs hatchability/fertility. The utilization of this artificial diet (S.A.D2) would supply the researchers with high-quality insects in adequate numbers, at specified times and specific stages of development for the bioassay, toxicological and biological studies.     

Consumption rate of some proteinic diets affecting hypopharyngeal glands development in honeybee workers

The experiment was carried out under laboratory condition to study the consumption of some proteinic diets and their effect on hypopharyngeal glands (HPG) development during nursing period. The results showed that the bee bread and the pollen loads mixture with sugar (1:1) were more consumed by honeybee workers followed by Nectapol® and Yeast-Gluten mixture. The lowest consumption amount was recorded with traditional substitute.Clear differences were found in HPG development under feeding with different diets. The maximum development degree was observed when fed with bee bread followed by pollen loads and mixture from Yeast, Gluten and sugar (1:1:2). The acinal surface of HPG showed clear difference under feeding with difference diets. The largest area was recorded when honeybee workers fed on bee bread followed by Yeast-Gluten-sugar mixture (diet,4) and pollen loads(diet,2).     

Field and semifield evaluation of impacts of transgenic canola pollen on survival and development of worker honey bees

A 2-yr field trial (2001 and 2002) and 1-yr semifield trial (2002) were conducted to evaluate the effect of transgenic herbicide (glyphosate) -tolerant canola Brassica napus L. pollen on larval and adult honey bee, Apis mellifera L., workers. In the field trial, colonies of honey bees were moved to transgenic or nontransgenic canola fields (each at least 40 hectares) during bloom and then sampled for larval survival and adult recovery, pupal weight, and hemolymph protein concentrations. No differences in larval survival, adult recovery, and pupal weight were detected between colonies placed in nontransgenic canola fields and those in transgenic canola fields. Colonies placed in the transgenic canola fields in the 2002 field experiment showed significantly higher hemolymph protein in newly emerged bees compared with those placed in nontransgenic canola field; however, this difference was not detected in the 2001 field experiment. In the semifield trial, bee larvae were artificially fed with bee-collected transgenic and nontransgenic canola pollen and returned to their original colonies. Larval survival, pupal survival, pupal weight, and hemolymph protein concentration of newly emerged adults were measured. There were no significant differences in any of the parameters measured between larvae that were fed transgenic canola pollen and those fed nontransgenic corn pollen. Results from this study suggest that transgenic canola pollen does not have adverse effects on honey bee development and that the use of transgenic canola dose not pose any threat to honey bees.     

Nutritional effects of supplementary diets on brood development,biological activities and honey production of Apis mellifera L

The present research work was conducted to assess the impact of nutrient-enriched diet on the physiological activities and subsequently honey yield. Eighteen colonies of Apis mellifera L. were selected from Dera Ismail Khan region, KPK, Pakistan, during the winter and summer seasons, 2019–2020. Five pollen supplement diets were prepared and provided to screen out the palatable one to be fed as pollen alternative nutrition to bee bread. Results of diet consumption regarding mean data for consumption rate displayed that soybean flour enriched artificial diet was maximally consumed (74.34 g) by honey bees per week. Minimum consumption was observed for grinded groundnut enriched diet (64.62 g) which was relatively lesser than the other tested artificial diets. Results of area of worker brood disclosed that soybean flour fortified diet (1489.27 cm²/colony) statistically noteworthy than the other artificial diets whereas control (463.51 cm²/colony) was least effective. Highest bee strength (10.00 bee frames/colony) was noted in the bee colonies fed with soybean flour fortified diet, date paste (8.0 bee frames/colony) was the next effective one, among the tested pollen replacement diets whereas relatively least (5 bee frames/colony) was noticed in case of grinded groundnut. Highest body weight (12.41 g) of neonate bees was noted in case of soybean enriched diet while lowermost (5.31 g) was noted in control bees. Results of wax cell built up and foraging efficiency were also superior in artificial diets than respective control bees. Hence, artificial diets especially soybean-enriched pollen alternative diet can boost up the physiology of honey bee leading to increased honey yield and profit.     

Recycled paper products as substitutes for gelling agent in screwworm (Diptera: Calliphoridae) larval diet

Absorbent recycled paper products were tested and compared with a sodium polyacrylate gelling agent, Aquatain, for solidifying larval diets of the screwworm, Cochliomyia hominivorax (Coquerel). A recycled newsprint product, Terra-Mulch, when mixed with water and dietary ingredients produced a diet medium of homogeneous texture that supported larval growth and development comparable to the standard diet prepared with the gelling agent. Biological yields as calculated from number of pupae per tray from both the diets were similar. Three other types of recycled paper products were tested. Although less homogeneous in texture, these diets were as good as the diet with Terra-Mulch. There were no significant differences in the mean larval and pupal weights, no. of pupae/tray, percent emergence, oviposition, percent hatch, and percent longevity. When tests were conducted through eight consecutive generations, no adverse effect was noticed in any of the life history parameters. Terra-Mulch is inexpensive and biodegradable compared with the gelling agent Aquatain that is expensive and not biodegradable. Replacing Aquatain with Terra-Mulch in the screwworm larval diet for mass rearing may result in an annual savings of more than half a million U.S. dollars in costs of material and labor.     

Behavior and molecular physiology of nurses of worker and queen larvae in honey bees (Apis mellifera)

In a honey bee colony, worker bees rear a new queen by providing her with a larger cell in which to develop and a large amount of richer food (royal jelly). Royal jelly and worker jelly (fed to developing worker larvae) differ in terms of sugar, vitamin, protein and nucleotide composition. Here we examined whether workers attending queen and worker larvae are separate specialized sub-castes of the nurse bees. We collected nurse bees attending queen larvae (AQL) and worker larvae (AWL) and compared gene expression profiles of hypopharyngeal gland tissues, using Solexa/Illumina digital gene expression tag profiling (DGE). Significant differences in gene expression were found that included a disproportionate number of genes involved in glandular secretion and royal jelly synthesis. However behavioral observations showed that these were not two entirely distinct populations. Nurse workers were observed attending both worker larvae and queen larvae, and there was no evidence of a specialized group of workers that preferentially or exclusively attended developing queens. Nevertheless, AQL attended larvae more frequently compared to AWL, suggesting that nurses sampled attending queen larvae may have been the most active nurses. This study serves as another example of the relationship between differences in gene expression and behavioral specialisation in honey bees.