Fine Structure of the Hemopoietic Tissues in the Mealworm Beetle, Tenebrio molitor

The fine structure of the hemopoietic tissue and its detailed reticular organization in the mealworm beetle, T. molitor were examined using light and scanning electron microscopes. The major hemopoietic tissues in the abdomen were located on the upper surface of the dorsal diaphragm which continuous over the ventral wall of the heart. Histologic characteristics of this hemopoietic tissues are dense clusters of cells. They are irregular in outline and are not surrounded by any connective tissue sheath. The hemopoietic tissue of this insect is consisted of three cellular components which are the reticular cells, hemocytic stem cells and several kinds of mature hemocytes. The reticular cells had numerous cytoplasmic processes and forming a complex network. The stem cells give rise to differentiating hemocytes of the different cell lineages. Mature hemocytes within this hemopoietic tissue are originated from the stem cells and differentiated into several types of hemocytes including prohemocytes, plasmatocytes, and granulocytes.     

Material Benefits from Multiple Mating in Female Mealworm Beetles (Tenebrio molitor L.)

In yellow mealworm beetles (Tenebrio molitor), females are sexually receptive throughout their adult lives. We examined how access to mates affected female fecundity by varying the number of matings per female and quantifying cumulative egg production. Also, we dissected females at successive intervals after a single mating to assess the relationship among time since mating, sperm supplies, egg load, and oviposition rate. Females that mated at intervals greater than 2 days did not produce as many eggs as females that mated every 2 days or were allowed to mate ad libitum. Dissections showed that the amount of sperm remaining in a female spermatheca was correlated with the number of eggs she had laid recently, which suggests sperm replenishment as the material benefit gained through multiple mating. However, females mate more frequently than necessary for sperm replenishment, and therefore material benefits alone may not fully explain the continuous receptivity of T. molitor females.     

黄粉虫胰蛋白酶样酶基因TMTLSP全长cDNA的克隆和序列分析

以黄粉虫(Tenebrio molitor)幼虫全RNA逆转录得到的cDNA为模板,参照地鳖（Eupolyphaga sinensis）纤溶酶（fibrinolytic enzyme）简并引物,进行温度梯度PCR．以得到的扩增产物为基础,采用RACE得到基因全长cDNA,命名为黄粉虫胰蛋白酶样丝氨酸蛋白酶(Tenebrio molitor trypsin-like serine protease,TMTLSP)．TMTLSP全长869 bp(GenBank No. JN662461),开放阅读框为777 bp,编码258个氨基酸,并具有蛋白酶样特有的起始位点、活性中心预计底物结合位点．经过比对分析,该基因编码的氨基酸序列与赤拟谷盗、谷蠹、光亮扁角水虻、美洲大蠊等多种昆虫的胰蛋白酶或丝氨酸蛋白酶有较高的相似性．本研究将为胰蛋白酶样丝氨酸蛋白酶的提取及研究提供更为广泛的材料及研究依据．     

Tenebrio molitor抗冻蛋白基因家族cDNA片段的克隆、序列分析及原核表达

利用反转录-多聚酶链式反应(RT-PCR)的方法, 克隆黄粉甲虫(Tenebrio molitor)抗冻蛋白基因cDNA片段并进行序列分析和原核表达。同源性分析表明, 获得9条新cDNA片段, 与黄粉甲虫抗冻蛋白基因家族的其他基因序列具有较高的同源性。重组质粒pGEX-4T-1-tmafp-XJ430, 转化E.coli BL21进行原核表达, SDS-PAGE分析结果表明, 抗冻蛋白基因以可溶性融合蛋白表达, 相对分子量为38 kDa。构建真核表达载体pCDNA3-tmafp-XJ430, 免疫小鼠, 获得的抗血清滴度为1:2 000。Western blotting 结果为单一的条带, 证明该抗血清具有针对抗冻蛋白TmAFP-XJ430抗原的专一性。     

The Site Specificity of Two Species of Gregarina in Tenebrio molitor Larvae

The site specificity of the apicomplexans Gregarina cuneata and Gregarina sleini , in larval Tenebrio molitor was investigated. Gregarina cuneata was found to inhabit the anteriormost region of the larval midgut, while G. steini was restricted to the posterior portion of the intestine. The site specificity of the pair was conserved in single and concurrent infections. Interspecific interactions do not seem to be presently responsible for the resource partitioning by the 2 gregarine species. Key words. Gregarina, site specificity, Tenebrio molitor.     

槲皮素对黄粉虫血淋巴酚氧化酶的生理效应

用酶标仪法测定了槲皮素对黄粉虫Tenebrio molitor酚氧化酶（phenoloxidase, PO）的生物活性。结果表明：在离体条件下,对黄粉虫血淋巴PO的IC₅₀为0.625 mmol/L。在活体情况下,当槲皮素-DMSO溶液或槲皮素水悬浮液（5 μL）注入虫体后,在浓度低于1.0 mmol/L时对黄粉虫血淋巴PO具有激活作用,当浓度高于2.0 mmol/L时则对PO具有明显的抑制作用;单独注射5 μL DMSO溶液对PO活性亦有一定的激活作用。试虫被注射槲皮素后,PO活性在2～4 h内迅速下降,然后缓慢上升,处理8 h左右达到最高点,其后低浓度处理PO活性保持不变,而高浓度处理则逐渐下降,提示低浓度槲皮素可以引起试虫的免疫反应。在PO测活体系中加入0.5％的BSA后对PO活性无影响,并能使槲皮素在测活体系中保持稳定。     

黄粉虫幼虫对硒的生物积累

在饲料中添加含硒化合物喂养黄粉虫Tenebrio molitor L.幼虫,测定幼虫硒含量、粪便硒含量和体重的变化,计算黄粉虫幼虫特定生长率及幼虫对硒生物积累系数,分析黄粉虫有效积累硒的条件。结果表明,饲料硒含量在15~20mg/kg时,幼虫硒含量明显提高,对硒的生物积累系数高于其它试验组水平,饲料硒含量过高,幼虫硒含量降低,正常生长受到抑制。黄粉虫幼虫特定生长率、取食量、排粪量、干物质含量随着饲料硒含量的增加而降低,死亡率、粪便硒含量随着饲料硒含量的增加而增大。饲料硒含量为15～20mg/kg时黄粉虫幼虫对硒的生物积累效果最好。     

泰山虫草对黄粉虫的侵染特性研究

报道了用泰山虫草分生孢子悬浮液接种活体黄粉虫,提取被侵染的黄粉虫体液,并测定不同侵染时间虫体液中酶活性的动态变化。结果表明:泰山虫草分生孢子能成功的侵染黄粉虫并长出子座;接种第2~4 d后即可导致黄粉虫体内活性发生不同程度的变化,说明黄粉虫具有独特的防御病原微生物的侵染机制。     

Enantiomerization and Enantioselective Bioaccumulation of Metalaxyl in Tenebrio molitor Larvae

The enantiomerization and enantioselective bioaccumulation of metalaxyl by a single dose of exposure to Tenebrio molitor larvae under laboratory condition were studied by high‐performance liquid chromatography tandem mass spectroscopy (HPLC‐MS/MS) based on a ChiralcelOD‐3R [cellulosetris‐tris‐(3, 5‐dichlorophenyl‐carbamate)] column. Exposure of enantiopure R‐metalaxyl and S‐metalaxyl in Tenebrio molitor larvae exhibited significant enantiomerization, with formation of the R enantiomers from the S enantiomers, and vice versa, which might be attributed to the chiral pesticide catalyzed by a certain enzyme in Tenebrio molitor larvae. Enantiomerization was not observed in wheat bran during the period of 21 d. In addition, bioaccumulation of rac‐metalaxyl in Tenebrio molitor larvae was enantioselective with a preferential accumulation of S‐metalaxyl. These results showed that enantioselectivity was caused not only by actual degradation and metabolism but also by enantiomerization, which was an important process in the environmental fate and behavior of metalaxyl enantiomers. Chirality 26:88–94, 2014. © 2013 Wiley Periodicals, Inc.     

利用黄粉虫分离土壤昆虫病原真菌

利用黄粉虫Tenebrio molitorL.作为寄主从土壤中诱感并分离昆虫病原真菌。结果显示,从吉林市的18个土样中分离出球孢白僵菌(Beauveria bassiana)、金龟子绿僵菌(Metarhiziumanisopliae)和玫烟色拟青霉(Paecilomyces fumosoroseus)。土样中的昆虫病原真菌检出率为77.78%。这表明利用黄粉虫分离土壤中的昆虫病原真菌是一种简单、有效的方法。     

黄粉虫对不同比例聚苯乙烯的取食效率研究

为探索黄粉虫Tenebrio molitor在降解聚苯乙烯过程中聚苯乙烯与麸皮的最佳配比,本研究将聚苯乙烯与麸皮按不同比例PS1(1:9),PS2(2:8),PS3(3:7),PS4(4:6),PS5(5:5),PS6(6:4),PS7(7:3),PS8(8:2),PS9(9:1),PS10(10:0),CK(0:10...     

利用黄粉甲繁育管氏肿腿蜂:不同蜂虫比对繁育效果的影响

利用黄粉甲Tenebriomolitor的蛹作为中间寄主繁育管氏肿腿蜂Sclerodermaguani,测定了不同蜂虫比对发育历期、寄生率等几项评价繁育效果的重要指标。结果显示接蜂时不同蜂虫比对雌蜂产卵量和育出的子代蜂数量有较明显的影响,当蜂虫比为2∶1时这2项重要指标较为理想。综合比较了寄生率、寄生成功率、子代蜂的发育历期、出蜂数、性比等各项指标后认为,人工扩繁时采用2∶1的蜂虫比较为适宜。     

黄粉虫在不同饲料比例下的泡沫降解率研究

为降低塑料泡沫的污染,探索在不影响黄粉虫Tenebrio molitor L.正常生长发育情况下的最大泡沫降解率。组建了黄粉虫在泡沫与麸皮的质量比为(1∶10),(1∶8),(1∶6),(1∶4),(1∶2)五种不同饲料配比下对泡沫的降解情况分析表,结果表明,黄粉虫在泡沫饲料比为 (1∶6)时泡沫的降解率最大,可达 74.21%,且此时的生长发育情况与空白对照组持平,而在泡沫饲料比为 1∶4 时只有 35.86% ,(1∶10),(1∶8),(1∶2)下分别为 45.58% ,49.26% ,37.24% 。泡沫降解率与虫的增重量都随泡沫占的比重的增加呈抛物线趋势,在(1∶6)时达最大,此后下降。分析得出结论,黄粉虫的泡沫饲料比为 1∶6 时既不影响黄粉虫生长且泡沫降解最优。      

Regulation of development of the grain beetle Tenebrio molitor by neuropeptides

Summary

Two neuropeptides (NPs), hydra head activator (HHA), and the luteinizing hormone releasing hormone (LHRH), were used to study their effects on the development of the larvae of the grain beetle Tenebrio molitor (mealworms). The larvae received the NPs in the course of development at the beginning or at the end of instar. The LHRH given at the beginning of the instar stimulated molting and pupation in larvae; it did not affect molting in the late period of the instar. The C-end fragment of LHRH_(9–10) produced fewer molts and increased the duration of the instar period. The effect of this fragment was weak when it was injected at the initial period of the instar; it was considerably stronger in the end of the instar. It inhibited development when administered during the last instar. On the contrary, the N-end fragment of the LHRH_(1–2) stimulated growth and development of larvae. The action of these NPs can be connected to the function of the hormones of metamorphosis. The HHA administrated to the larvae at different days of the instar stimulated molting but in the last instar it delayed pupation. The C-end of the HHA_(7–11) stimulated molting when injected during the whole period of the instar. The other C-end fragments, the HHA_(8–11) and HHA_(9–11) had inhibitory effects on the molting. The injection of the N-end fragment of the HHA_(1–5) had no effect. The action of HHA and its fragments did not correlate with the function of the hormones of metamorphosis.     

黄粉虫防卫物质苯醌的测定及其去除

采用分光光度法测定了黄粉虫的防卫毒素苯醌的含量。结果表明 :各虫态各性别的黄粉虫在不同发育阶段其苯醌含量均有不同 ,各虫态的苯醌含量随日龄而增大 ,雌蛹和雌成虫的苯醌含量均高于同一日龄的雄蛹和雄成虫。经刺激后各虫态的苯醌含量升高 2 0 72 %～ 41 64%,刺激后用水清洗的虫体其体内苯醌含量下降 88 2 7%～ 92 60 %;烘烤也可使苯醌大量挥发和分解 ,采用刺激 -水洗 -研磨 -烘烤工艺处理幼虫可清除 94 3 0 %苯醌 ,此法可为保障黄粉虫作为饲用昆虫的安全性提供依据     

不同温度条件对利用黄粉甲繁育管氏肿腿蜂的影响

在恒温 2 0 ,2 2 ,2 4,2 6,2 8和 3 0℃的实验条件下 ,利用黄粉甲TenebriomolitorL .做中间寄主繁育管氏肿腿蜂SclerodermaguaniXiaoetWu ,测定了其发育历期和寄生率等生物学参数。结果显示 ,在被测温度范围内 ,管氏肿腿蜂的发育速率与温度呈正比关系 ,即随着温度的升高 ,发育历期逐渐缩短。卵至羽化的发育起点温度为 1 3 60℃ ,有效积温为 3 2 0 0 7日·度。温度对管氏肿腿蜂的寄生率、寄生成功率、产卵量、子代蜂的数量和性比都有较显著影响。结合发育历期的结果 ,认为适于人工大量繁蜂的温度范围为 2 4～ 2 8℃ ,2 6℃为最适温度。     

黄粉虫及其肠道微生物对聚氯乙烯的生物降解作用

【背景】塑料废物的累积是越来越严重的环境污染问题,其中聚氯乙烯(polyvinyl chloride,PVC)作为最常用的塑料之一,其生产与消耗量极大。【目的】基于黄粉虫(Tenebriomolitor)幼虫自然取食塑料的现象,探究黄粉虫及其肠道微生物对聚氯乙烯类塑料的生物降解作用。【方法】通过观测黄粉虫取食聚氯乙烯过程的重量变化、傅里叶变换红外光谱(fouriertransforminfrared spectroscopy,FTIR)、黄粉虫肠道微生物的筛选和高通量测序等方法研究黄粉虫幼虫及其肠道微生物对聚氯乙烯的降解作用。【结果】研究结果表明:仅以PVC作为碳源类营养来源,每200条黄粉虫为一组,共3组,喂养32d后,每组平均取食了0.499±0.023gPVC,黄粉虫平均体重增长0.015±0.002 g。采用傅里叶变换红外光谱检测发现虫粪组分中PVC官能团中的C-C骨架峰明显减弱,表明PVC长链有断裂现象。高通量测序黄粉虫幼虫肠道微生物菌群的结果显示肠道菌群以哈夫尼菌属(Hafnia)、摩根氏菌属(Morganella)、大肠埃希氏杆菌属(Escherichia coli)和未...     

黄粉虫幼虫小分子量抗茵肽的分离纯化

昆虫抗菌肽具有良好的抑菌效果,有望开发成新一代抗生素。本文以金黄色葡萄球菌和大肠杆菌混合液作为诱导源,采用针刺法使黄粉虫TenebriomolitorL．幼虫感染微生物产生抗菌肽,并对抗菌肽进行了提取、色谱分离纯化及抑菌活性检测。结果显示,诱导组和对照组的三氟乙酸粗提物无抑菌活性;经SephadexG50、SuperdexPeptide凝胶色谱分离后,从诱导组和对照组均可获得对革兰氏阳性菌金黄色葡萄球菌、枯草芽孢杆菌有抑菌作用的组分,而且诱导组活性明显高于对照组;通过Resource15RPC反相色谱分离纯化,从诱导组获得一具有明显抑制革兰氏阳性菌的组分,质谱检测该组分为混合肽,主要由分子量为1876．21u、1904．21u的小肽组成,可能是一种比Thanatin分子量更低的昆虫抗菌肽。