白星花金龟成虫肠道细菌的分离鉴定

目的自星花金龟是我国北方常见腐食性昆虫,为了利用其腐食性行为转化处理有机垃圾、畜禽粪便及农作物秸杆等农业有机废弃物资源,并探索其中肠消化的微生态机制,而对其肠道细菌进行系统研究。方法从白星花金龟成虫肠道中无菌操作分离获得5株细菌,对其菌体形态、培养性状、染色反应、生理生化反应等进行鉴定及数量测定。结果白星花金龟成虫肠道的5个细菌菌株分别属于气球菌属（Aerococcus）、短状杆菌属（Brachybacterium）、棍状杆菌属（Clavibacter）、李斯特菌属（Listeria）、皮杆菌属（Dermabacter）,以皮杆菌属和棍状杆菌属数量最多,分别为1．3×10^7和1．0×10^7;其次为短状杆菌属和气球菌属,分别为8×10^6和5×10^6;李斯特菌属数量最少,为7×10^5。结论各菌株之间细菌数量存在明显差异,以皮杆菌属细菌数量最多,分离未获得真菌和放线菌。     

白星花金龟室内外繁育技术研究

[目的]根据新疆气候特点和白星花金龟Protaetia brevitarsis Lewis生物学特性,探究在新疆人工繁育白星花金龟的可行性及潜力,以期为白星花金龟在新疆的人工繁殖和转化利用畜禽粪便提供科学依据.[方法]以牛粪为繁育基质,在适宜季节分别在室内外保育白星花金龟的蛹及作为成虫产卵基质,最终统计比较室内外成虫的羽化及产卵情况.[结果]室外露天条件更利于白星花金龟的羽化,老熟幼虫做土室26 d后羽化出成虫,比室内及室外棚下处理提前4d.在化蛹30d后进入羽化高峰期,持续13d,羽化率83％.成虫雌雄比接近1∶1,雌虫羽化时间早于雄虫1-3 d.白星花金龟成虫室内外产卵量差异不显著(P＜0.05),室内单雌产卵量109.80粒,室外105.94粒,成虫羽化后30-35 d进入产卵高峰期,第60天室内外成虫产卵量显著下降(P＜0.05),甚至不产卵.[结论]白星花金龟规模化人工繁育在新疆是可行性的,在室外自然条件和室内控温条件下白星花金龟繁育的效果差异不显著.     

白星花金龟绿僵菌分离及毒力测定

目的 2008年在山东泰安市黄庄堆肥中发现白星花金龟幼虫被病原菌寄生死亡,病虫体上长满灰绿色霉层,对其进行分离,经纯化获得1株真菌,编号为M-08。通过对幼虫接种,证明其致病性。方法对分离纯化出的菌株M-08,根据菌体形态特征、培养性状,按常规方法鉴定。结果 M-08菌株鉴定结果为金龟子绿僵菌长孢型变种Metarhiziumnaisopliae[Metsch Sorokin Var major（Johnston）Tulloch],毒力测定结果表明,对白星花金龟幼虫致死率高达93.3%~100%。结论金龟子绿僵菌长孢型毒力测定表明具有很高的毒性,今后有良好的应用前景。     

白星花金龟捕食落叶松毛虫蛹

白星花金龟Potosia brevitarsis Lewis属鞘翅目金龟甲科的一种杂食性昆虫。近年来作者在对落叶松毛虫的调查中发现,白星花金龟对该害虫种群数量的调控起着不可忽视的作用。     

不同补水方式对白星花金龟水分平衡的影响

为研究水分散失和水分补充对新疆新害虫白星花金龟Potosia brevitarsis Lewis的影响,在30℃恒温条件下,采用重量法测定白星花金龟脱水过程以及不同补水方式下(补蒸馏水;补盐水;补糖水)体内水分含量的变化,并与步甲、拟步甲的水分代谢进行比较。结果表明,白星花金龟的脱水与拟步甲科的网目拟地甲Opatrum subaratum Fald相似,在10h内脱水率均约为5%,而斑步甲Anisodactylus signatus 10h内脱水率约为20%。不同补水处理后,白星花金龟体内含水量迅速增加,随后继续脱水;脱水10d后,补蒸馏水、补盐水和补糖水处理的白星花金龟脱水率分别为28%,27%和21%,而未补水白星花金龟的脱水率为34%;未补水处理的甲虫脱水率和补蒸馏水和补盐水处理之间无显著性差异,而和补糖水处理之间存在显著性差异。未补水、补蒸馏水、补盐水和补糖水处理的LT50分别约为9,12,13和17d,补糖水能有效延长甲虫的存活时间。糖能有效地增加白星花金龟体内含水量,对维持其体内水分平衡起重要作用。     

白星花金龟对寄主植物挥发物的电生理反应

[目的]白星花金龟Protaetia brevitarsis是我国经济上重要的农业害虫.本研究拟鉴定苹果、葡萄和玉米3种果实在两种状态(健康与受白星花金龟成虫为害)下的挥发物中能引起白星花金龟雌成虫触角电生理反应的挥发物成分.[方法]采用动态顶空吸附法收集苹果、葡萄和玉米3种果实的挥发物,用气相色谱-触角电位联用技术(...     

不同农业有机废弃物对白星花金龟生物学特性影响研究

【目的】白星花金龟Protaetia (Liocola) brevitarsis (Lewis)幼虫取食农作物秸秆、畜禽粪便等农业废弃物,在自然界中发挥着净化环境、变废为宝的作用。【方法】本文选用8种农牧业有机废弃物,包括腐熟沼渣、平菇菌糠、猪粪、牛粪、玉米秸秆、25%玉米秸秆+牛粪、50%玉米秸秆+牛粪、75%玉米秸秆+牛粪作为饲料,研究不同农业有机废弃物对白星花金龟生物学特性的影响。【结果】沼渣组和平菇菌糠组单雌产卵量较高,分别为130.17粒和117.00粒,显著高于猪粪、牛粪和玉米秸秆处理;在牛粪与玉米秸秆混配组,各处理单雌产卵量为101.50-108.67粒,不同混合比例间无显著差异。从幼虫孵化率比较,平菇菌糠组和猪粪组幼虫孵化率较高,分别为87.78%和82.22%;玉米秸秆及其与牛粪不同比例混合处理的幼虫孵化率均很低,仅为46.67%-57.78%。从幼虫化蛹率、羽化率比较,玉米秸秆添加不同比例牛粪处理组的化蛹率和羽化率均较高,分别达到90.00%-95.56%和88.89%-91.11%;沼渣、猪粪等单一物质处理的化蛹率和羽化率均较低,分别低于90%和70%。单一物质处理中,沼渣、平菇菌糠、牛粪、玉米秸秆处理组羽化后均为雌虫比例大于雄虫比例。玉米秸秆与牛粪混配组中,仅50%玉米秸秆+牛粪处理组为雌虫比例大于雄虫比例;雌、雄成虫的体长及体宽均在(17-19)mm、(10-11)mm之间,沼渣、猪粪和牛粪处理组体长略小。以不同农业有机废弃物饲喂的白星花金龟全世代发育历期中,牛粪饲喂组白星花金龟历期最短,为(315.29±0.55)d;沼渣饲喂组白星花金龟历期最长,为(330.75±1.76)d。【结论】不同农业有机废弃物均可作为白星花金龟繁育基质,并可为其幼虫转化处理农业有机废弃物提供理论依据。     

产卵基质模式和性比对白星花金龟生殖力的影响

[目的]为探究产卵基质模式和雌雄比对白星花金龟生殖力影响,寻求提高白星花金龟繁殖力的技术参数.设计复合产卵基质模式的思想,可为其他产卵于固体基质中的昆虫提高生殖力提供借鉴.[方法]本研究设计了5种产卵基质模式(牛粪、细沙单一基质、粪沙混、粪沙分和粪液沙复合基质)和5个雌雄比(早∶♂=4∶8、5∶7、6∶6、7∶5和8∶4),在室外敞口棚和自然温光条件下,开展对白星花金龟成虫产卵量和死亡率、幼虫孵化率的影响研究.[结果]5种产卵基质下白星花金龟成虫产卵量的顺序为粪沙分＞粪沙混＞牛粪＞粪液沙＞细沙,其中,白星花金龟在粪沙分复合产卵基质40 d的单雌产卵量达129.0粒,是原生产卵基质——牛粪的1.72倍;5个雌雄比处理比较,白星花金龟群体产卵量的顺序为7∶5＞8∶4＞6∶6＞5∶7＞4∶8;单雌产卵量的顺序为4∶ 8＞6∶6＞5∶7＞7∶5＞8∶4,最高达3.60粒/d.2项试验下成虫死亡率和幼虫孵化率在各处理间无显著差异,幼虫孵化率均在90％以上.综合分析和结合群体实际雌雄比可知,雌雄比为1∶1时,对于保障白星花金龟的群体繁殖力效果最佳.[结论]牛粪和细沙两复合体系作为产卵基质和维持雌雄比1∶1,可显著提高白星花金龟的繁殖力.     

白星花金龟人为条件生物学与应用潜力

本文选取人工饲养的东亚飞蝗虫砂作为白星花金龟Potosia brevitarsis Lewis幼虫饲料、成虫产卵基质进行人为条件下的生物学研究。测定了其在不同温度条件下各虫态发育历期、不同食物下成虫单雌产卵量、不同酵化周期条件下东亚飞蝗虫砂2龄、3龄幼虫的转化力;分析了幼虫虫体和虫砂的主要营养成分;实施了虫砂盆栽小白菜试验。结果显示,白星花金龟适宜的繁育温度为22~28℃,25℃全世代发育历期为126.62(±1.05)d;以11种水果为食料,成虫单雌产卵量顺序为:芒果>菠萝>葡萄>苹果>桃>梨>西瓜>甜瓜>李子>杏>西红柿,最高207粒,最低68粒;东亚飞蝗虫砂酵化周期与取食龄期的最佳对应组合分别为2龄30 d和3龄25 d,折合每转化100 g可增长2龄虫体3.04 g、产生虫砂81.90 g或3龄虫体4.37 g、虫砂74.48 g;3龄幼虫含蛋白质(53.8%)、脂肪(6.6%)、18种氨基酸(44.68%),是昆虫源高蛋白原料;虫砂含有机质(18.72%)和氮磷钾(5.95%)。盆栽小白菜试验表明,1 cm^2施用0.13 g,效果最佳。水果残余物作为白星花金龟成虫食物,东亚飞蝗虫砂作为成虫产卵基质和幼虫繁育饲料,保持适宜温光条件,可以实现白星花金龟周年繁育。白星花金龟虫体和虫砂在生态循环农业、医药等领域具有应用潜力。     

苹毛丽金龟对植物挥发物成分的触角电位及行为反应

为筛选适宜的苹毛丽金龟Proagopertha lucidula (Faldermann)引诱剂化学物质, 对已报道的金龟子科两性引诱剂单一组分进行了触角电位（EAG）和Y型嗅觉仪中的嗅觉反应测定。结果表明: 在10 μg刺激剂量下测试的11 种植物挥发物中, 只有顺-3-己烯-1-醇、香叶醇、茴香脑和丁香酚有明确的EAG反应值, 其他7种化合物具有弱反应值。剂量反应曲线表明, 具有明确的EAG活性的4种化合物激发的反应强度与化合物的剂量呈正相关。在Y型嗅觉仪中, 顺-3-己烯-1-醇、香叶醇、茴香脑和丁香酚4种化合物在1, 10, 100 μg剂量下的行为反应百分率都显著高于对照溶剂正己烷, 且雌雄之间无显著差异。该研究结果可为开发苹毛丽金龟田间引诱剂奠定基础。     

Hepatoprotective and antineoplastic properties of Protaetia brevitarsis larvae

Protaetia brevitarsis larvae have been used as a traditional medicine to treat various liver diseases. Freeze‐dried powder and solvent partitioned fractions of Protaetia brevitarsis larvae, in this study, were examined for hepatoprotective and antitumor activities in vivo and in vitro. Oral administration of the powder reduced signs of acute and chronic liver injuries in diethylnitrosamine‐induced hepatotoxic mouse model. Various cancer cell types, but not normal primary hepatocytes, were killed by the hexane, ethyl acetate and butanol fractions, which were partitioned from larval extracts. Apoptosis and autophagy were primarily induced in cancer cells by hexane and ethyl acetate fractions, respectively, and can be the cytotoxic mechanisms of the fractions. Further analyses of the insect fractions could lead to identification of novel bioactive substances that can be exploited as therapeutics for various human liver diseases.     

Inhibition of platelet aggregation and thrombosis by indole alkaloids isolated from the edible insect Protaetia brevitarsis seulensis (Kolbe)

Protaetia brevitarsis seulensis (Kolbe) has been temporarily registered as a food material by the Ministry of Food and Drug Safety of Korea (MFDS). The current study aimed to discover small antithrombotic molecules from this edible insect. Five indole alkaloids, 5‐hydroxyindolin‐2‐one ( 1 ), (1R,3S)‐1‐methyl‐1,2,3,4‐tetrahydro‐β‐carboline‐3‐carboxylic acid ( 2 ), (1S,3S)‐1‐methyl‐1,2,3,4‐tetrahydro‐β‐carboline‐3‐carboxylic acid ( 3 ), (3S)‐1,2,3,4‐tetrahydro‐β‐carboline‐3‐carboxylic acid ( 4 ) and L‐tryptophan ( 5 ), were isolated from the insect. Among them, compounds 1 and 2 prolonged aPTT and PT and impaired thrombin and FXa generation on HUVEC surface. Moreover, these compounds inhibited platelet aggregation. Antithrombotic effects of compounds 1 and 2 were further confirmed in pre‐clinical models of pulmonary embolism and arterial thrombosis. Collectively, these results demonstrated that compounds 1 and 2 could be effective antithrombotic agents and serve as new scaffolds for the development of antithrombotic drug.     

华山松木蠹象聚集信息素分离鉴定和引诱效果

为了研制华山松木蠹象Pissodes punctatus Langor et Zhang的引诱剂,对华山松木蠹象的虫粪和雄虫后肠挥发性物质进行了分析鉴定和室内外生物活性测试。经GC-MS测定,发现在华山松木蠹象的新鲜虫粪和雄虫后肠挥发性物质中,除了松树挥发性的单萜烯如α-蒎烯、β-蒎烯和3-蒈烯以外,还存在1-甲基-2-异丙烯基-环丁烷乙醇（grandisol）。室内Y-型嗅觉仪趋向实验表明,较低浓度的3-（+）-蒈烯、1-甲基-2-异丙烯基-环丁烷乙醇及其相应的醛1-甲基-2-异丙烯基-环丁烷2醛（grandisal）,引起华山松木蠹象的正趋向反应。林间引诱试验表明,1-甲基-2-异丙烯基-环丁烷乙醇和1-甲基2-异丙烯基-环丁烷2醛对华山松木蠹象具有一定的引诱作用。由此推断,1-甲基-2-异丙烯基-环丁烷乙醇可能是华山松木蠹象的集结信息素成分之一。     

白星花金龟(Protaetia brevitarsis)幼虫抗菌物质的分离纯化

以药用昆虫白星花金龟(Protaetia brevitarsis)幼虫为研究对象,通过Sephadex G-50、RP-HPLC等技术分离纯化天然抗菌活性物质.经初步鉴定,该系列抗菌活性物质对枯草芽孢杆菌(Bacillus subttilis)表现出较强的抑菌活性.用MALDI-TOF MS对样品进行分析,检测到相对分子质量分别为2254.045 9、2622.4438的两种抗菌活性物质.推测这些抗菌活性物质在白星花金龟幼虫抵御微生物的侵袭中起到重要作用.     

The effects of feed materials on the nutrient composition of Protaetia brevitarsis larvae

To find out the effect of feed materials on the nutrient composition of Protaetia brevitarsis larvae (Pbl), three different feed materials were prepared and fed to the Pbl populations. After the Pbl population reached the third instar stage, they were boiled and dried for the nutrient analysis. The results showed that the nutrient composition of Pbl varied in response to the feed materials. Therefore, it is critical to set a feed material standard to guarantee the efficacy of Pbl medicinal property.     

Characterization of the Hemocytes in Larvae of Protaetia brevitarsis seulensis: Involvement of Granulocyte-Mediated Phagocytosis

Hemocytes are key players in the immune response against pathogens in insects. However, the hemocyte types and their functions in the white-spotted flower chafers, Protaetia brevitarsis seulensis (Kolbe), are not known. In this study, we used various microscopes, molecular probes, and flow cytometric analyses to characterize the hemocytes in P. brevitarsis seulensis. The circulating hemocytes were classified based on their size, morphology, and dye-staining properties into six types, including granulocytes, plasmatocytes, oenocytoids, spherulocytes, prohemocytes, and adipohemocytes. The percentages of circulating hemocyte types were as follows: 13% granulocytes, 20% plasmatocytes, 1% oenocytoids, 5% spherulocytes, 17% prohemocytes, and 44% adipohemocytes. Next, we identified the professional phagocytes, granulocytes, which mediate encapsulation and phagocytosis of pathogens. The granulocytes were immunologically or morphologically activated and phagocytosed potentially hazardous substances in vivo. In addition, we showed that the phagocytosis by granulocytes is associated with autophagy, and that the activation of autophagy could be an efficient way to eliminate pathogens in this system. We also observed a high accumulation of autophagic vacuoles in activated granulocytes, which altered their shape and led to autophagic cell death. Finally, the granulocytes underwent mitotic division thus maintaining their number in vivo.     

Purification and Properties of Ferritin from the Last Instar Larval Hemolymph of Protaetia brevitarsis (Coleoptera)

Ferritin, an iron storage protein, has been purified from the last larval hemolymph of Protaetia brevitarsis (coleoptera) by KBr density gradient ultracentrifugation and resource Q (anion exchange chromatography) using fast performance liquid chromatography (FPLC) system. The iron content of ferritin was determined by atomic emission spectroscopy and FereneS stain. Ferritin of P. brevitarsis is shown to have two different subunits presented on a SDSPAGE in normal (N) and ironinjected (I) hemolymph. SDS PAGE revealed that the ferritin consists of two major polypeptides of 27 and 28 kDa in normal hemolymph. Interestingly, however, 30 kDa subunit was substituted for 28 kDa when iron was injected into the hemolymph. Apporximate isoelectric points of 27 kDa, 28 kDa, and 30 kDa ferritin subunits were 6.7, 6.75, and 6.8, respectively. Ferritin of P. brevitarsis was detected by FereneS stain and confirmed by Western blotting using its polyclonal antibody. Other characteristics such as amino acid composition and Nterminal amino acid sequence were investigated. Amino acid composition of ferritin (N and I) was rich in alanine, glycine, glutamine or glutamic acid and serine, but poor in histidine, arginine, methionine and phenylalanine.