熊蜂营养与饲料研究进展

熊蜂是重要的商业传粉昆虫之一,人工驯养和应用熊蜂传粉是弥补自然传粉蜂锐减,满足现代农业授粉需求的重要举措。营养和饲料是实现熊蜂驯养和工厂化生产的关键问题之一。本文综述了单一和混合花粉饲料,不同蛋白质和脂肪比例的饲料,不同氨基酸水平的饲料,以及碳水化合物饲料中添加营养元素或者蜂王浆等对熊蜂发育的影响。其中,混合花粉相比单一花粉对幼虫体重发育有促进作用;欧洲地熊蜂Bombus terrestris成年蜂摄取的蛋白质和脂肪的最佳比例为14∶1;高浓度的氨基酸饲料对蜂王产卵和幼虫成熟有促进作用;碳水化合物中添加营养元素能够促进蜂王产卵,蜂群成群;饲料中添加蜂王浆能够提高蜂王存活率和产卵率。建议未来以本土熊蜂的生产群体为研究对象,针对熊蜂阶段性营养需求及可能影响熊蜂发育的肠道共生菌等因素开展研究。     

云南短头熊蜂的繁育特性研究

短头熊蜂Bombus breviceps是云南省优势蜂种,为更好地挖掘本土熊蜂资源,繁育农业授粉蜂群,本研究从云南3个地区（屏边、个旧和昆明）收集短头熊蜂蜂王后,在相同条件下饲养,统计分析其蜂王产卵前期、工蜂发育期、蜂群生长特性、群势、产卵蜂王率及蜂群可应用率等繁育特性指标。结果表明：3个地区工蜂的发育期相同,屏边地区的蜂王产卵前期最长,蜂群始见工蜂时间及工蜂数量达到6头、30头和60头的时间也最长,均显著高于个旧地区和昆明地区（P<0.05）。3个地区短头熊蜂的群势差异不显著（P>0.05）,但昆明地区的工蜂数量最少,显著低于屏边地区和个旧地区（P<0.05）,而子代蜂王数量显著高于屏边地区和个旧地区（P<0.05）,雄性蜂数量显著高于屏边地区（P<0.05）。昆明地区的产卵蜂王率和蜂群可应用率高于屏边地区和个旧地区。总体而言,云南的短头熊蜂群势强,产卵蜂王率75%以上,蜂群可应用率均在63%以上,易于人工繁育,具有重要的授粉利用价值。     

熊蜂气管内寄生螨布赫纳蝗螨的生物学特性观察

对新疆与河北地区野生明亮熊蜂及小峰熊蜂蜂王内寄生螨布赫纳蝗螨Locustacarus buchneri的生物学特性进行了初步观察。结果表明： 该螨对两种熊蜂的寄生率分别为10.43%和8.3%,寄生部位为气管与气囊。室内饲养至30天的蜂王体内寄生螨量明显高于饲养至60天的蜂王。未被螨寄生的蜂王与被寄生者比较,前者体重显著高于后者,但二者间产卵始期、产卵率、食卵率和存活率差异不显著。     

不同饲料对小峰熊蜂工蜂群发育的影响

糖和花粉对熊蜂的生长发育和繁殖起重要作用。本研究测定和分析了2种糖源（白砂糖、蜂蜜）和5种花粉（杏花粉、油菜花粉、向日葵花粉、玉米花粉、茶花粉）的饲料组合对小峰熊蜂Bombus hypocrita的无王工蜂群寿命、产卵前期时间、产卵量、幼虫拖出数、卵杯数、雄蜂出房时间和子代雄蜂数量的影响。结果表明： 只饲喂糖类食物时, 工蜂寿命显著短于有花粉的饲料组, 而且工蜂不产卵。含有花粉的各饲料组之间工蜂寿命存在显著差异, 饲喂玉米花粉的工蜂寿命小于其他4种花粉组。在工蜂产卵前期时间和卵杯数量方面, 10种含有花粉的饲料组之间差异不显著; 但在产卵量、幼虫拖出数、雄蜂出房时间和子代雄蜂数量等方面, 各饲料组之间差异较大。白砂糖和杏花粉组的产卵量最高; 杏花粉组和茶花粉组的幼虫拖出数量显著低于其他花粉组; 油菜花粉组和杏花粉组的雄蜂出房时间显著短于其他花粉组; 杏花粉组的子代雄蜂数量显著高于其他饲料组。结果显示不同饲料组合对小峰熊蜂工蜂群的发育影响很大。因此建议在熊蜂的人工繁育过程中, 在不同的发育阶段应给予不同的饲料配方。     

不同食料对野蚕黑卵蜂卵巢发育和卵子发生的影响

以20%浓度蜂蜜、蔗糖、葡萄糖、果糖、甘露糖为食料,研究了它们对野蚕黑卵卵巢发育和卵子发生的影响。结果表明,与对照相比(蒸馏水),蜂蜜、蔗糖、葡萄糖和果糖能促进野蚕黑卵蜂雌蜂卵巢发育和卵子发生,可使其卵巢管中较高的成熟卵量维持较长时间,即可延缓该蜂的卵子重吸收。甘露糖对野蚕黑卵蜂的卵子形成有一定的促进作用,但其所起的作用显然不如蜂蜜及其它3种糖类,并且也不能延缓该蜂的卵子重吸收。     

棉铃虫卵巢形态与卵子发生过程观察

害虫发生高峰期、 发生量的准确预测和田间防治适期的确定与种群雌虫卵巢结构及卵子发生过程密切相关。为了明确棉铃虫Helicoverpa armigera卵巢结构及卵子发生过程, 本研究利用光学体视显微镜和透射电子显微镜, 对棉铃虫成虫卵巢管和卵子的超微结构进行了研究, 并确定了发育级别划分标准。结果表明: 根据卵巢的形状、 卵的产生过程、 卵黄沉积情况等将棉铃虫卵巢发育程度分为6个级别, 即发育初期(0级)、 卵黄沉积前期(Ⅰ级)、 卵黄沉积期(Ⅱ级)、 成熟待产期(Ⅲ级)、 产卵盛期(Ⅳ级)和产卵末期(Ⅴ级)。根据卵子发生过程中滋养细胞、 卵母细胞的变化, 将卵子发生期分为3个阶段： 卵黄发生前期、 卵黄发生期和卵黄成熟期。本研究首次对棉铃虫的卵子发生进行电子显微观察, 并完善了棉铃虫卵巢发育的分级标准, 为进一步研究棉铃虫的生殖发育机理提供了理论参考, 对田间棉铃虫种群发生期和发生量的预测预报也有重要的实践参考价值。     

兰州熊蜂蜂王头部产卵相关microRNA生物信息学分析

【目的】头部是熊蜂感觉和取食中心,也是生理行为的指挥中心。microRNA（miRNA）参与重要生理行为的调控。本研究初步探索了产卵和未产卵兰州熊蜂Bombus lantschouensis蜂王头部差异表达miRNA及其靶基因,以期探明蜂王头部miRNA在产卵过程中作用。【方法】以本土优良蜂种兰州熊蜂 B. lantschouensis 为材料,利用Solexa高通量测序技术对产卵和未产卵蜂王头部miRNA进行测序,运用生物信息学软件对miRNA及其靶基因进行预测;并应用qPCR技术验证极显著差异表达的miRNA。【结果】共获得兰州熊蜂产卵和未产卵蜂王头部miRNA的clean data数,分别为14 228 864和21 431 031 条reads;长度分析表明,在19~24 nt序列中22 nt序列数量最多,分别占产卵和未产卵蜂王序列的45.8%和45.1%。miRNA预测结果显示,共获得297个miRNA,其中270个为已知miRNA,有92个存在于蜜蜂中,其余178个分别存在于蜜蜂以外的其他物种中;另外27个为新的miRNA。在蜜蜂92个已知的miRNA中,在产卵蜂王头部中表达的共有91个,特异表达的有2个;在未产卵蜂王头部中表达的共有90个,特异表达的有1个。27个新的miRNA中,在产卵蜂王头部中表达的共有22个,特异表达的有2个;未产卵蜂王头部中表达的共有25个,特异表达的有5个。miRNA差异表达分析表明,在产卵与未产卵蜂王头部,共有8个已知miRNA表达量达到差异极显著水平（P<0.01）。qPCR结果表明,这8个表达差异极显著的miRNA均存在于熊蜂体内。在8个差异极显著的miRNA中,3个miRNA进行靶基因预测,发现它们共同调控9个靶基因,这些基因主要参与GTP结合和转录调控。【结论】本研究首次利用高通量测序技术鉴定了熊蜂头部组织的miRNA,并发现蜂王产卵与否受到miRNA差异表达的调控。结果为今后深入开展熊蜂miRNA功能验证及产卵调控机制研究奠定了基础。     

美洲斑潜蝇成虫的卵巢结构观察及营养影响卵子发生的试验

卵育型(Synovigenic)植食性昆虫的雌成虫需要取食寄主植物组织或非寄主食物(如花蜜、花粉等)以延长寿命和促进产卵。为了明确寄主食物和非寄主食物对美洲斑潜蝇Liriomyza sativae卵子发生的功能差异,本实验在明确美洲斑潜蝇成虫卵巢发育动态和结构的基础上,比较了清水(饥饿,对照)、菜豆叶片、20%葡萄糖溶液、菜豆叶片+20%葡萄糖溶液四种处理下的雌成虫卵子发生动态。结果显示:美洲斑潜蝇为强卵育型昆虫,初羽化成虫体内无卵子;两侧卵巢管数之和为11-18根,大多数情况下,左右两侧卵巢管数目不等。成虫在饥饿条件下,几乎不能形成成熟卵子;继续取食寄主叶片、20%葡萄糖溶液和寄主叶片+20%葡萄糖溶液,均可显著促进卵子发生和卵成熟,但补充葡萄糖溶液起到极显著促进作用;成虫不具备卵子重吸收功能。本文重点从卵育型角度,对美洲斑潜蝇取食寄主食物和非寄主食物的营养功能进行了讨论。研究结果为美洲斑潜蝇以及该类植食性害虫的室内饲养和田间生态调控提供了依据。     

CO2麻醉对中华蜜蜂处女蜂王卵巢发育的影响

【目的】明确中华蜜蜂Apis cerana cerana蜂王羽化后生殖系统的动态变化,探索CO₂麻醉对中华蜜蜂处女蜂王卵巢发育的影响。【方法】通过人工育王技术和蜂王的储存获取不同日龄的中华蜜蜂处女蜂王,观察蜂王羽化后生殖系统的形态,测定生殖系统各项形态学指标(体重、卵巢重、受精囊直径和卵巢管数目)。中华蜜蜂处女蜂王连续2 d(分别于5和6日龄时)接受不同浓度(50%, 75%与90%)CO₂麻醉8 min, 以及不同日龄处女蜂王接受75% CO₂麻醉1~3次(每次8 min),待蜂王发育至10日龄时,利用石蜡切片技术与HE染色法观察蜂王卵巢形态、测定蜂王卵巢重和蜂王卵巢最大卵母细胞长度,通过实时荧光定量PCR(RT-qPCR)测定10日龄蜂王头部卵黄原蛋白基因(Vg)的相对表达量。【结果】中华蜜蜂蜂王的卵巢管数目与受精囊直径不会随着日龄发生明显的波动,但蜂王卵巢重在6-12日龄明显增加,之后将趋于稳定。与对照组(未接受CO₂麻醉处理)相比,50%, 75%与90% CO₂麻醉后中华蜜蜂蜂王卵巢重与最大卵母细胞长度均增加,并且头部Vg基因的相对表达量上调。其中CO₂浓度为75%,麻醉次数2次显著促进了中华蜜蜂蜂王的卵巢发育。【结论】中华蜜蜂蜂王羽化后卵巢会发生明显的发育变化,而CO2麻醉能加速卵巢的发育。     

地熊蜂蜂群发育性状评价及其饲料花粉配比优化

【目的】为了明确熊蜂蜂群发育过程中具有代表性的性状指标及其饲料花粉最优配比。【方法】以山杏Armeniaca sibirica花粉、山柳Salix caprea花粉和油菜Brassica rapa花粉为原料,通过{3,3}混料设计得到7种配比的混合花粉,分析不同混合花粉对地熊蜂Bombus terrestris(L.)无王工蜂群的产卵前时间、幼虫拖出数、幼虫总数量、幼虫总重量、蛹总数量、蛹总重量、雄蜂出房时间、出房雄蜂数量和雄蜂出生重9个蜂群性状指标的影响,使用主成分分析对蜂群性状指标进行综合评价,并通过混料回归模型预测熊蜂蜂群饲养过程中的最优花粉配比。【结果】地熊蜂无王工蜂群发育过程的9个性状指标可以归纳为幼虫发育、蛹发育、成蜂发育、蜂群发育周期4类评价因子,其中幼虫总数量、蛹总重量、雄蜂出生重、产卵前时间和雄蜂出房时间是5个主要性状指标;以蜂群主要性状指标为评价依据,得出蜂群饲养过程中最优花粉配比:当以油菜花粉单独饲喂蜂群时蜂群产卵前时间短、幼虫总数量最多、蛹总重量最大,当山杏花粉、山柳花粉和油菜花粉以1︰1.5︰1.5比例饲喂蜂群时雄蜂出房时间最短,当山柳花粉和油菜花粉以3︰1比例饲喂蜂群时雄蜂出生重最大。【结论】明确了地熊蜂无王工蜂群发育过程中的重要性状指标和饲料花粉最优配比,为进一步研究商品化熊蜂群不同发育阶段的营养需求奠定了基础。     

Diapause survival and post-diapause performance in bumblebee queens (Bombus terrestris)

Diapause survival and post-diapause performance (i.e., if a queen starts to lay eggs) of in total 2210 bumblebee queens (Bombus terrestris) were measured under different diapause regimes: 5 temperatures (–5, 0, 5, 10 and 15 °C) in combination with 5 durations of exposure (1, 2, 4, 6 and 8 months).The results show that weight at the start of diapause determines to a large extent whether a queen will be able to survive diapause. Queens with a wet weight below 0.6 g prior to diapause did not survive, but for those queens exceeding this threshold a higher pre- diapause weight did not increase their post-diapause performance.There was no effect of temperature on diapause survival; 76% of the variance in survival could be explained by the duration of the treatment. Neither temperature nor duration of exposure had an effect on post-diapause performance. The preoviposition period of the queens that laid eggs was also determined. The preoviposition period was affected by both temperature and duration of exposure: the preoviposition period decreased with decreasing temperature but also with increasing duration of the treatment.     

Effect of different narcosis procedures on initiating oviposition of prediapausingBombus terrestris queens

Four experiments aimed at the stimulation of starting oviposition were carried out with bumblebee queens (Bombus terrestris L.) from colonies belonging to the ecotype of Central Western France and reared in a glasshouse. After mating, queens were narcotized with carbon dioxide, confined singly in small boxes (11×5×4.5 cm) and kept in a dark room at 28–29 °C and 60%–65% r.h. They were fed on a sugar solution and a pollen-syrup mixture. No effects were discernible if the narcosis was applied 20 to 30 days after mating instead of 5 days, nor if the queens were submitted to a 4 to 5 day period at 34°C following narcosis. Survival rates ranged from 65% to 68 %. If the queens were reared under fluorescent tubes (L8∶D16) after narcosis the mean delays to egg-laying were significantly reduced compared to a dark treatment (21 days instead of 39), as was their variability (s.e.=1.6 day instead of 3.1 days). The survival rates were respectively 73% and 67%. Under the same photoperiod (L8∶D16) the CO₂ narcosis repeated at a 24h interval had the same efficacy whether its duration was 10 min or 5 min. The delays to egg-laying were respectively 20 days (s.e=1.5) and 25 days (s.e.=4.8) with survival rates close to 73%. Egg-laying could also be induced in non-narcotized queens with a survival rate of 54% and delays to oviposition close to those of queens narcotized 2×10 min.     

明亮熊蜂繁育室内印度谷斑螟的形态特征与生物学特性

明亮熊蜂Bombus lucorum L.是一种重要的温室果菜传粉昆虫,印度谷斑螟Plodia interpunctella(Hbner)是危害明亮熊蜂繁育的主要害虫之一。2年的研究结果表明,印度谷斑螟的形态特征与危害蜜蜂巢房的大蜡螟Gallerie mellonellaL.和小蜡螟Achroia grisella Fabricius明显不同。印度谷斑螟以饲喂熊蜂的花粉为载体传播进熊蜂繁育室,在熊蜂繁育室内可以连续繁殖,1年发生6～8代,且世代重叠。印度谷斑螟幼虫主要以蜂群内的剩花粉为食,当花粉不足时,就开始取食巢房和蜂蛹,对熊蜂的规模化繁育危害较大,每年的5～8月、11～2月为危害高峰期。印度谷斑螟的发育受环境和食物的影响很大,在温度为28℃、相对湿度为60%熊蜂繁育室内,印度谷斑螟的卵期为4d,幼虫期为19～23d,蛹期为11d,成虫期为4～20d。成虫羽化后3～4h即可进行交配,雌蛾交配后当天就开始产卵,产卵期4～10d不等,平均产卵量107.8粒。在熊蜂繁育室消毒期间,食物短缺,印度谷斑螟幼虫进入休眠状态,各龄幼虫均可发生休眠现象。印度谷斑螟不危害蜜蜂,在蜜蜂巢房中不能存活。     

Efficacy of exemestane,a new generation of aromatase inhibitor,on sex differentiation in a gonochoristic fish

We report the first use of exemestane (EM), a steroidal aromatase inhibitor (AI) commercially known as aromasin, in studies of sex differentiation in fish. The effectiveness of EM was examined in two different age groups of the gonochoristic fish, Nile tilapia (Oreochromis niloticus). Untreated control fish (all female) showed normal ovarian differentiation through 120 days after hatching (dah), whereas fish treated with EM at 1000 and 2000 µg/g of feed from 9 dah through 35 dah, the critical period for sex differentiation, exhibited complete testicular differentiation; all stages of spermatogenic germ cells were evident and well developed efferent ducts were present. Fish treated with EM at 1000 µg/g of feed from 70 dah through 100 dah significantly suppressed plasma estradiol-17β level and increased level of 11-ketotestosterone. Furthermore, untreated control fish showed strong gonadal expression of the steroidogenic enzymes P450 cholesterol-side chain-cleavage enzyme (P450scc), 3β-hydroxysteroid dehydrogenase (3β-HSD), and cytochrome P450 aromatase (P450arom). In contrast, EM-treated fish showed immunopositive reactions against P450scc and 3β-HSD but not against P450arom in interstitial Leydig cells. These results indicate that treatment of tilapia juveniles with EM during sex differentiation leads to the development of testes, apparently by a complete suppression of aromatase activity.     

Bumblebees at an alpine site in northern Sweden: temporal development, population size, and plant utilization

The bumblebee community in an alpine valley in northern Sweden was studied for two seasons. Only two species of bumblebees, Bombus alpinus and B. hyperboreus. are common at the site. Bombus hyperhoreus is an obligate nest parasite on B. alpinus : the latter species is four times more abundant than the former. The number of captures on any given day was very low. However, the number of queens foraging in the area was estimated by a mark-reacapture method to ca 350 specimens, indicating that the queens forage over large areas. The dates for capturing queens carrying pollen in their corbiculae and the emergence of workers and males indicate that the queens only have time to produce one batch (or possibly two overlapping batches) of workers. The utilization of different plant species corresponds well with their flowering phenology at the site. The unspecialized flowers of Saxifraga oppositifolia are the main pollen and nectar source in the early part of the season. The bumblebees switch to Astragalus alpinus and Bartsia alpina as soon as they come into flower: these species show a typical bee pollination syndrome, the corbicular loads most commonly consisted of 91-100% of pollen from a single species. The change in flower utilization and the composition of the corbicular loads give no support for the hypothesis that aretie bumblebees should be more generalistic in their foraging than temperate species. The preference for Astragalus and Bartsia will probably have an effect on outcrossing in late-flowering specimens of Saxifraga oppositifolia , which has a very extended flowering period.     

尖唇散白蚁工蚁和繁殖蚁卵子发生比较

为探讨白蚁工蚁品级性腺不育的原因,采用组织学观察与测量对尖唇散白蚁Reticulitermes aculabialis Tsai et Hwang的工蚁和繁殖蚁的卵子发生各阶段进行比较和分析。结果表明,成熟工蚁的卵子发生与繁殖蚁相比,仅有卵母细胞的分化期和生长期,没有卵黄形成期,其分化期和生长期的卵母细胞大小与繁殖蚁这2个时期的卵母细胞大小呈显著性差异;生长期时,工蚁的卵母细胞和滤泡细胞之间出现较大间隙并且呈现萎缩退化状态;工蚁卵子发生相当于雌性繁殖蚁末龄若虫水平。该结果将为工蚁不能生殖提供组织学上的证据,同时又表明了工蚁具有潜在生殖能力的结构基础。     

一日龄饥饿对粘虫成虫卵巢发育和飞行能力的影响

为了明确羽化后0～24 h (1日龄)饥饿处理对迁飞型粘虫Mythimna separata (Walker)生殖系统和飞行系统发育的影响,对1日龄饥饿处理后粘虫的卵巢发育级别、产卵前期、飞行肌和飞行能力进行了系统研究。结果表明, 1日龄饥饿对粘虫成虫卵巢发育级别、产卵前期、中胸背纵肌干重、飞行能力等均有显著的影响。饥饿后成虫卵巢发育级别显著高于对照,产卵前期显著短于对照。成虫中胸背纵肌干重从饥饿后96 h开始下降,至120 h时饥饿粘虫(5.1 mg)显著低于对照(5.86 mg)。成虫飞行能力从饥饿后96 h开始显著低于对照,此时饥饿处理粘虫蛾的飞行距离平均为22.03 km,飞行时间平均为4.07 h,显著低于同时期的对照粘虫的飞行距离(42.3 km)和飞行时间(7.99 h),尽管在饥饿后72 h内差异不显著。实验结果显示,1日龄饥饿可以显著抑制粘虫飞行系统的发育,加速飞行向生殖的转变。     

The most effective pollinator revisited: pollen dynamics in a spring-flowering herb

Most flowers are visited by a wide range of potential pollinators. However, their efficiency in pollen removal and deposition, and other behavioural factors affecting pollination effectiveness may greatly differ among taxa, and even individuals. Fritillary (Fritillaria meleagris L., Liliaceae) is a spring-flowering, critically endangered plant in the Polish flora, red-listed in most of the European countries of its range. Based on indirect evidence, that is, body pollen loads, visitation frequency and seasonal abundance, it is estimated that its key pollinators are queen bumblebees, but, as shown in the literature, the largest Fritillaria pollen loads are carried by solitary bees. To study pollinator effectiveness for floral visitors to F. meleagris, we performed a garden experiment, where we analysed pollen deposition and assessed pollen removal per single flower-visit in the plant. Similarly to field conditions reported in the literature, our experimental plants were serviced by nectar-seeking bumblebee queens and two taxa of solitary bees, small pollen-collecting Andrena and large, nectar-seeking Anthophora males. When “quality” component was addressed, despite the character of visits, insects from all groups deposited more pollen than was found on unvisited flowers, but they did not differ significantly from each other in pollen deposition on virgin stigmas. We also found some differences in pollen removal both within- and among-visitor species and control flowers, unfortunately due to extremely high variation of the results they were all statistically insignificant. However, when “quantity” component of insect performance was concerned, we observed that over 81 % of visits were by bumblebees. Bombus queens stayed on flowers significantly less time than small Andrena individuals (13 % of recorded visits) and equally long as Anthophora males (only 6 % of visits). We conclude that although all the visitor groups can pollinate the flowers of F. meleagris, bumblebee queens indeed proved to be the most effective pollinators of the plant, when both quality and quantity components of pollination are concerned.