Enantioselective Bioaccumulation of Diniconazole in Tenebrio Molitor Larvae

The enantioselective bioaccumulation of diniconazole in Tenebrio molitor Linne larva was investigated with liquid chromatography tandem mass spectrometry based on the ChiralcelOD‐3R[cellulose tri‐(3,5‐dimethylphenyl carbamate)] column. In this study we documented the effects of dietary supplementation with wheat bran contaminated by racemic diniconazole at two dose levels of 20 mg kg^‐1 and 2 mg kg^‐1 (dry weight) in Tenebrio molitor. The results showed that both doses of diniconazole were taken up by Tenebrio molitor rapidly in the first few days, the concentrations of R‐enantiomer and S‐enantiomer at high doses reached the highest level of 0.55 mg kg^‐1 and 0.48 mg kg^‐1, respectively, on the 1^st d, and the concentrations of them obtained a maxima of 0.129 mg kg^‐1 and 0.128 mg kg^‐1 at low dose, respectively, on the 3^rd d, which means that the concentration of diniconazole was proportional to the time of achieving the highest accumulated level. It afterwards attained equilibrium after a sharp decline at both 20 mg kg^‐1 and 2 mg kg^‐1 of diniconazole. The determination results from the feces of Tenebrio molitor demonstrated that the extraction recovery (ER) values of the high dose group were higher than that of the low dose group and the values were all above 1; therefore, it could be inferred that enantiomerization existed in Tenebrio molitor. Additionally, the biota accumulation factor was used to evaluate the bioaccumulation of diniconazole enantiomers, showing that the bioaccumulation of diniconazole in Tenebrio molitor was enantioselective with preferential accumulation of S‐enantiomer. Chirality 25:917–922, 2013. © 2013 Wiley Periodicals, Inc.     

Bioaccumulation and Excretion of Enantiomers of Myclobutanil in Tenebrio molitor Larvae Through Dietary Exposure

The bioaccumulation and excretion of enantiomers of myclobutanil in Tenebrio molitor larvae through dietary exposure under laboratory conditions were investigated using high‐performance liquid chromatography tandem mass spectrometry (HPLC‐MS/MS) based on a ChiralcelOD‐3R [cellulosetris‐tris‐(3, 5‐dichlorophenyl‐carbamate)] column. The wheat bran fed to Tenebrio molitor larvae was spiked with racemic myclobutanil at two dose levels of 20 mg/kg and 2 mg/kg (dry weight). The results showed that there was a significant trend of enantioselective bioaccumulation in the larvae with a preferential accumulation of (?)‐myclobutanil in 20 mg/kg dose exposure, but it was not obviously observed in the 2 mg/kg dose group. A kinetic model considering enantiomerization between the two enantiomers based on first‐order reactions was built and the rate constants were estimated to discuss the kinetic reason for the different concentrations of individual enantiomers in the larvae. The approximations implied an inversion between the two enantiomers with a relatively higher rate of the inversion from (?)‐myclobutanil to (+)‐myclobutanil. Meanwhile, analysis of data of excretion samples suggested the active excretion is probably an important pathway for the insect to eliminate myclobutanil rapidly with nonenantioselectivity as a passive transport process, which was consistent with the low accumulation efficiency of myclobutanil measured by BAF (bioaccumulation factor). Chirality 25:890–896, 2013. © 2013 Wiley Periodicals, Inc.     

Enantioselective Bioaccumulation and Dissipation of Soil‐Associated Metalaxyl Enantiomers in Tubifex

Many pesticides are chiral compounds and stereochemistry is an important factor for any reaction of chiral structures in biological systems. In this study, experiment about bioaccumulation of the two metalaxyl enantiomers in Tubifex (Oligochaeta, Tubificida) was conducted in laboratory aquatic ecosystems. Terrestrial soil spiked with two dose levels of metalaxyl was employed as the artificial bottom substrate. A method of determination of metalaxyl enantiomers in tubifex tissue, soil and overlying water were developed by HPLC. During a 14‐day exposure, concentrations of metalaxyl in tubifex increased with the of soil concentration, however, the enantioselective bioaccumulation was only detected at high‐dose exposure group, with the preferential accumulation of (‐)‐(R)‐metalaxyl. The bioturbation activity of tubifex decreased water clarity and released soil‐associated metalaxyl to overlying water. In those experiments where tubifex was exposed to metalaxyl from soil, pore water and overlying water, each route contributed to the total body burden, and our results indicated the pore water and soil are the primary exposure routes for high‐dose exposure concentration treatment. Chirality 26:33–38, 2013. © 2013 Wiley Periodicals, Inc.     

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。     

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.     

饲养密度对黄粉虫幼虫生长发育的影响

黄粉虫原是一种仓库害虫 ,由于其幼虫、蛹、成虫都含有丰富的蛋白质和多种氨基酸 ,已被作为饲用资源昆虫来开发利用。作为一种群集性的昆虫 ,黄粉虫幼虫的生长受密度影响可分为 2个阶段 ,一是幼虫孵化后 1个月这段时间内 ,高密度处理的幼虫平均体重大于低密度处理的幼虫平均体重 ;二是孵化 1个月后直到化蛹 ,这段时间低密度的幼虫生长要比高密度的幼虫快得多。密度越大 ,历期越长。但密度对化蛹率没有影响。在幼虫生长过程中 ,各处理组中只有 3 2 0头虫密度处理组的幼虫在生长 1个月后显出拥挤效应。     

黄粉甲幼虫抗菌物质的诱导及其抗菌活性

采用饥饿法、紫外线照射法和针刺法处理黄粉甲Tenebriomolitor 6龄幼虫后均能诱导其 产生抗菌物质,收集的血淋巴上清液对真菌有抑制作用,对细菌无抑制作用;经热处理后的血 淋巴上清液则对细菌有抑制作用,而对真菌无抑制作用。SDS-PAGE检测结果发现,与未诱导的 对照相比经诱导的黄粉甲幼虫血淋巴中,原有的一类大分子蛋白质如分子量分别为97kD、44 kD和37 kD左右的蛋白质缺失;而ESI-MS分析结果显示诱导后比诱导前黄粉甲幼虫血淋巴中有 小分子物质产生,推测可能是此类缺失蛋白质分解为小分子量的抗菌肽,从而表现出抗菌活性 。     

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．     

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。     

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。     

Cyfluthrin and Deltamethrin Induce Changes in the Fat Body Composition of Tenebrio molitor Larvae,Males and Females

The purpose of present study was to analyze the chemical composition of the fat body of insects (Tenebrio molitor) and its eventual variation after the application of insecticides. Using the GC/MS technique, the fat body composition of larvae, males and females was analyzed. The insects were stressed with selected insecticides containing cyfluthrin (Cyflok 50EW) and deltamethrin (K‐othrine) recommended for this species of insect by insecticides producers and farmers. The effect of these two chemicals on the chemical compounds patterns from fat body of insects was compared. Possible differences in the fat body composition between developmental stages and the sex of insects were also compared. Possible identified compounds included amino acids, carboxylic acids, alcohols, sterols, glycerol and urea. Our results show that the composition of lipids was different depending on the stage of development, the insecticide used and the elapsed time since its application.     

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。     

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...     

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。      

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     

Physical and chemical properties of primary defences in Tenebrio molitor

Prevention and reaction are the foundation for any defence system. In insects, the primary defences against pathogens and parasites limit invasion; the secondary ones (e.g. immune system) act when the cuticle and other primary defences fail. Because investment in both aspects of defence may be costly, they should be regulated in a plastic or variable way in accordance with the risk of infection. The mealworm beetle Tenebrio molitor L. changes cuticle colour and its resistance to fungal infection when subject to high population density, although such resistance is a result of the primary (cuticle) defences rather than the secondary (immunological) ones. The present study tests the hypothesis that the physical and chemical properties of the primary defences in T. molitor change with cuticular darkness. Beetles expressing black phenotypes (or with darker cuticle) have a thicker cuticle, with four well organized layers (epi‐, exo‐, endocuticle and formation zone) and more melanin than tan beetles. The cuticle properties investigated in the present study are likely to be the underlying mechanisms of pathogen resistance in black beetles, including the content of carbonylated proteins, which in black beetles was almost half that of tan beetles after exposure to ultraviolet radiation. It is proposed that, in polyphenic insects (such as mealworm beetles), primary and secondary defences are regulated pleiotropically, with the genes responsible for the expression of one defence having a positive effect on others, whereas, in polymorphic insects, there is no such link and so investment in one defence may impair others.     

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.