取食重金属铜对棕尾别麻蝇亲代及子代生长发育与繁殖的影响

为了评估取食含重金属铜饲料对棕尾别麻蝇Boettcherisca peregrine亲代及子代的生长发育与繁殖的影响,在室内给棕尾别麻蝇初产幼虫饲喂含不同浓度（200, 400,800和1 600 µg/g）Cu²⁺的饲料直至化蛹,并对亲代和子代的生长发育和繁殖有关指标进行了观察和分析。结果表明：低浓度的Cu²⁺（200 µg/g）对其体重和体长起促进作用,但对幼虫历期、化蛹率、蛹历期、羽化率、性比、交配率和产仔量无显著作用;较高浓度的Cu²⁺ 则有抑制作用,且Cu²⁺处理浓度越高,亲代幼虫、蛹和雌雄成虫的体重越轻,幼虫和蛹的体长越短,化蛹率、羽化率、交配率和产仔量越低,幼虫历期和蛹期越长,成虫寿命越短。但Cu²⁺处理对成虫性比则无显著的影响。相比之下,经Cu²⁺处理后雌虫所产的子代若不再经Cu²⁺处理,其子代生存、生长发育与繁殖则基本不受影响,说明Cu²⁺对亲代的影响不能遗传至子代。此外,还探讨了该蝇亲代与子代体内Cu²⁺含量在其变态过程中的变化。     

幼虫期染镉对棕尾别麻蝇繁殖力及生殖腺发育的影响

通过在饲料中添加重金属镉(Cd)饲喂棕尾别麻蝇Boettcherisca peregrine的幼虫,研究镉对其成虫繁殖能力和生殖腺发育的影响.结果表明,棕尾别麻蝇幼虫在取食含100 μg/g、200 μg/g、400 μg/g和800 μg／g Cd2的人工饲料后,成虫的羽化率和寿命都明显下降;但镉处理对成虫的交配率无...     

镉和铜离子诱导棕尾别麻蝇幼虫脂质过氧化丙二醛动态变化分析

采用饲料含毒法,测定了棕尾别麻蝇Boettcherisca peregrine 1日龄幼虫连续取食含不同浓度Cd^2＋或Cu^2＋的人工饲料,体内丙二醛含量的变化过程。实验结果表明：二种重金属离子能诱导昆虫的脂质过氧化反应,并且存在浓度依赖效应和时间依赖效应。在0—1600μg/g饲料浓度范围内,丙二醛（malondialdehyde,MDA）含量与幼虫取食Cd^2＋和Cu^2＋的浓度呈线性关系,所求回归方程分别为y=0．2789＋0．0002x和y=0．3141＋0．0002x。当处理浓度为800μg/g时,在0-120h时间内,MDA含量与幼虫取食Cd^2＋和Cu^2＋的时间呈线性关系,回归方程分别为y=0．3041＋0．0042x和y=0．3251＋0．0029x。     

棕尾别麻蝇雄性附腺分泌物的生理功能

棕尾别麻蝇Boettcherisca peregrina Robinean-Desvoidy雄性附腺的分泌物中包含许多生物活性分子,将成熟的雄性附腺分泌物注射到未交配的雌虫体内,发现其能影响雌虫生殖活性的很多方面:如降低雌虫的再次交配率;增加卵巢内的含卵量;缩短交配后雌虫的寿命。另外,还证明附腺分泌物中存在有可抑制革兰氏阳性菌的物质的存在。     

重金属Cd2+对棕尾别麻蝇血细胞数量、包囊作用和形态结构的影响

为了揭示重金属对昆虫细胞免疫的影响及其机理,本文采用血细胞计数、台盼蓝染色、延展与包囊率测定, 以及显微与超微结构观察等方法,以经重金属Cd²⁺（浓度为150 μg/g）处理的棕尾别麻蝇Boettcherisca peregrina及其对照为研究对象,分别测定了处理组与对照组血细胞的数量与存活率、延展与包囊率,观察其形态结构,并比较了两组间的差异。结果表明： 棕尾别麻蝇初产幼虫经连续喂饲含Cd²⁺的饲料24,48,72和96 h后,与对照相比其血细胞总数和存活率显著下降,而血细胞的延展率和包囊作用的显著下降则分别出现于取食72和48 h之内,此后则下降不显著;由Cd²⁺处理幼虫发育形成蛹的血细胞包囊作用也显著低于对照。形态结构观察结果表明,Cd²⁺处理对幼虫的原血细胞、浆血细胞、颗粒血细胞和类绛色血细胞的显微形态影响不大,但可导致部分浆血细胞不能产生伪足;但均能导致各类血细胞的超微结构发生不同程度的变化,其中主要变化包括：细胞膜受损或破裂,染色质呈现凝聚, 线粒体和内质网等细胞器明显减少,以及胞质内出现空囊泡。结果说明重金属Cd²⁺对棕尾别麻蝇的血细胞可具有毒害作用。     

棕尾别麻蝇幼虫肠道溶纤活性蛋白酶的分离纯化和性质

从以富含纤维蛋白的血凝块为食物的棕尾别麻蝇幼虫肠道浸提液中分离纯化出3种具有溶纤活性的蛋白酶,分别命名为BPGFP1,BPGFP2和BPGFP3。其中,BPGFP1由两个分子量分别为32000和30000的亚基组成。BPGFP2和BPGFP3均为单体,分子量分别为40000和28000。这三种蛋白酶具有相似的底物特异性和抑制剂特性。三种蛋白酶均能降解溶纤活性蛋白酶的特异底物纤维蛋白,Chromzym,P,Chromzym UK和S－2288。三种酶还能够强烈降解类胰蛋白酶专一底物Bz-Phe-Val Arg NA,cBz Gly-Pro-Arg NA,Bz-Pro－Phe-Arg NA和Bz-Val-Gly-Arg NA.PMSF,STI,LBTI和SBBI能够对三种蛋白酶活怀有极强的抑制作用。三种溶纤活性蛋白酶均在pH9.0-10.0范围内表现出较高活性。     

一株棕尾别麻蝇胚胎细胞系的建立及其特性分析

双翅目昆虫细胞系广泛应用于遗传学、发育生物学、分子生物学、人和动物体病原学以及昆虫抗微生物肽的研究。本研究建立了一株新的棕尾别麻蝇Sarcophaga peregrina胚胎细胞系。该细胞系的原代培养始于2008年11月17日, 取材于棕尾别麻蝇晚期胚胎组织, 在Shields & Sang M3昆虫培养基中于28℃恒温培养, 在第26天进行第1次传代, 至今已历时21个月, 传代72次, 生长状态稳定, 被命名为Sp-E-HNU11。该细胞系的细胞形态主要呈梭形和近圆形, 杂以少量巨型细胞, 紧密贴壁生长。细胞群体倍增时间为42 h。染色体数目一般为10条或12条, 为二倍体或亚二倍体细胞系; 除一对颗粒状微型染色体外, 其他染色体呈短杆状。细胞系的β-萘酯酶和谷草转氨酶同工酶谱上分别显示出1条和3条酶带。随机引物扩增多态性 (random amplified polymorphic DNA, RAPD) 分析结果显示, 该细胞系与小菜蛾细胞系Px-E-HNU12、草地贪夜蛾细胞系IPLB-Sf-9和家蚕细胞系Bm-21E-HNU5呈现明显不同的带型特征。 Sp-E-HNU11细胞系的建立为昆虫抗微生物肽及其他相关的研究工作增添了新的研究工具和生产载体。     

免疫亲和层析法纯化棕尾别麻蝇(Sarcophagaperegrina)幼虫血淋巴凝集素

 报道了利用免疫亲和层析法纯化棕尾别麻蝇幼虫血淋巴凝集素的结果．哺乳动物红细胞能够特异地吸附凝集素．用兔红细胞与麻蝇幼虫血淋巴凝集素形成的复合体免疫供血家兔,得到麻蝇幼虫血淋巴凝集素的抗体．再利用抗体制备亲和吸附柱,通过免疫亲和层析一次性纯化了麻蝇幼虫血淋巴凝集素． Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ结果显示,该凝集素的分子量约为７３ ｋ Ｄ．这一结果,与用对麻蝇幼虫血淋巴凝集素有抑制作用的糖蛋白—胎球蛋白和甲状腺球蛋白为配基,亲和层析纯化的结果完全相同,表明用这种免疫亲和层析法纯化凝集素是可行的．为不清楚专一性识别糖或专一性识别糖不典型,难于用普通亲和层析纯化的凝集素,提供了一种有效的纯化方法．     

棕尾别麻蝇(Sarcophaga peregrina)幼虫与蛹血淋巴凝集素的纯化与特性

用亲和层析法纯化了棕尾别麻蝇幼虫和蛹血淋巴凝集素。以兔红细胞吸附幼虫血淋巴凝集素为抗原制备的抗体、球球蛋白和甲状腺蛋白等三种亲和层析吸附剂纯化得到的幼虫凝集素是相同的,其分子量73kD左右。用甲状腺球蛋白为亲和配基纯化的蛹血淋巴凝集素由二种亚基组成,其分子量分别为30和32kD。幼虫和蛹血淋巴凝集素活性的抑制糖明显不同：乳糖、岩藻糖和N－乙酰半乳糖胺对幼虫血淋巴凝集素活性有抑制作用;而甘露糖胺、半乳糖胺和葡萄糖胺则对蛹血淋巴集素有一定抑制。而且,用兔红细胞吸附幼虫血淋巴凝集素为抗原制备的抗血清对蛹的凝集素活性无交叉反应,表明这两种凝集素是不相同的。虽然本文所纯化的麻蝇蛹血淋巴凝集素的分子量和Komano等报道的麻蝇蛹以及幼虫体壁 伤害诱导的凝集素SPL相同,但其糖的抑制特性有明显差异。     

Conversion of Solid Organic Wastes into Oil via Boettcherisca peregrine (Diptera: Sarcophagidae) Larvae and Optimization of Parameters for Biodiesel Production

The feedstocks for biodiesel production are predominantly from edible oils and the high cost of the feedstocks prevents its large scale application. In this study, we evaluated the oil extracted from Boettcherisca peregrine larvae (BPL) grown on solid organic wastes for biodiesel production. The oil contents detected in the BPL converted from swine manure, fermentation residue and the degreased food waste, were 21.7%, 19.5% and 31.1%, respectively. The acid value of the oil is 19.02 mg KOH/g requiring a two-step transesterification process. The optimized process of 12∶1 methanol/oil (mol/mol) with 1.5% H₂SO₄ reacted at 70°C for 120 min resulted in a 90.8% conversion rate of free fatty acid (FFA) by esterification, and a 92.3% conversion rate of triglycerides into esters by alkaline transesterification. Properties of the BPL oil-based biodiesel are within the specifications of ASTM D6751, suggesting that the solid organic waste-grown BPL could be a feasible non-food feedstock for biodiesel production.     

Resistance selection with cadmium and changes in the activities of antioxidases in Boettcherisca peregrina (Diptera: Sarcophagidae)

In order to establish a physiological link between antioxidases and the resistance level of insects to cadmium (Cd), natural populations of Boettcherisca peregrina (Diptera: Sarcophagidae) were maintained for 20 generations and reared either on an uncontaminated diet or on a diet contaminated with cadmium (Cd) at a concentration equivalent to the median lethal concentration (LC₅₀) as determined every five generations. A relatively susceptible strain (S) and a Cd-resistant strain (R) were selected. The metal accumulation, growth and development, reproduction, and antioxidant enzyme activities in these strains were analyzed. The results showed that R-strain organisms had enhanced juvenile survivorship, increased Cd accumulation, and increased adult female fecundity when compared with S-strain. The larval enzyme activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione S-transferase (GST) in R-strain larvae were higher than those in S-strain larvae when fed diets with or without Cd. This indicates that Cd resistance in B. peregrina larvae is mediated by SOD, CAT, GR, and GST.     

棕尾别麻蝇金属硫蛋白的分离纯化及性质分析

将棕尾别麻蝇Boettcherisca peregrina幼虫置于含800 μg/g CdCl₂的食物中取食48 h后,可诱导金属硫蛋白(MT)的合成。诱导处理后的幼虫匀浆上清液经Sephadex G-50分子筛柱、UNO^TM Q1阴离子交换柱和Bio-Gel P-6脱盐柱层析,纯化得到2个亚型,即MT-Ⅰ和MT-Ⅱ。MT-Ⅰ和MT-Ⅱ的分子量均为9 kD,每蛋白分子均含7个Cd和20个巯基,且具254 nm的Cd-SH特征吸收肩。两者的氨基酸组成中,以半胱氨酸含量最高,分别为36.6%和31.8%;而芳香族氨基酸和组氨酸含量甚少,约1%～2%。     

Accumulation of cadmium and its effects on growth, development and hemolymph biochemical compositions in Boettcherisca peregrina larvae （Diptera： Sarcophagidae）

Newly emerged larvae of the fleshfly, Boettcherisca peregrina were exposed to two different CdCl2 concentrations of 100μg/g and 400 mg/g diet fresh weight （DFW）. They were administered in the diets until the end of larval stage. Cd-exposed larvae accumulated significant amounts of Cd and this accumulation increased with the exposure dose and time. The body weights were lightened and lengths of larvae were shortened considerably after Cd exposure, especially at the higher Cd concentration. The total larval duration was also extremely affected due to Cd exposure. The average duration was prolonged significantly by 14 h at the lower Cd concentration, while it was increased by 33.7 h over controls at the higher Cd concentration. A significant decrease in contents of either soluble proteins, total lipids or caloric values in the hemolymph occurred due to Cd exposure throughout the entire tested period but after 120 h of Cd exposure. In contrast, when exposed to Cd with its higher concentration, total sugar contents in the hemolymph were increased strikingly over the whole tested time, except after 96 h of Cd exposure, while they were not apparently altered except after 24 h of Cd exposure at the lower concentration. Thus, it is suggested that Cd exposure shows significant adverse impact on the growth and development, as well as metabolism, in larvae of this fleshfly, depending on its exposure time and dose.     

丽蝇蛹集金小蜂寄生对寄主蛹可溶性蛋白与芳基蛋白组成与含量的影响

为了探讨蛹期寄生蜂对寄主蛋白代谢的寄生生理效应,利用Bradford蛋白含量测定法、Western免疫印迹法及酶联免疫吸附检测法研究了棕尾别麻蝇Boettcherisca peregrina蛹被丽蝇蛹集金小蜂Nasonia vitripennis寄生后其脂肪体和血淋巴中可溶性蛋白及芳基蛋白组成与含量的变化。结果表明：寄生蛹脂肪体和血淋巴中可溶性蛋白的组成与未寄生相比基本无明显差异; 不论寄生与否寄主蛹脂肪体和血淋巴中芳基蛋白亚基分子量均为80 kDa,该亚基在脂肪体中未出现降解现象,而在血淋巴中仅于寄生后12 h的寄主蛹中呈现2条分子量相近的Western免疫印迹带,说明其降解可能先于未寄生对照。就含量而言,寄生蛹脂肪体中可溶性蛋白含量除寄生后24 h外均显著低于未寄生对照,芳基蛋白含量除寄生后48 h外也均显著低于未寄生对照,其中寄生后12 h的含量仅为未寄生的32.0%。寄生蛹血淋巴中可溶性蛋白含量多低于未寄生蛹,且寄生后2,12,24 h的差异达显著水平;芳基蛋白的含量均有低于未寄生的趋势,其中寄生后12 h的含量为未寄生的17.0%。综合认为,丽蝇蛹集金小蜂的寄生可导致寄主脂肪体和血淋巴中可溶性蛋白及芳基蛋白含量下降。     

Geographic variation in the incidence of pupal diapause in Asian and Oceanian species of the flesh fly Boettcherisca (Diptera: Sarcophagidae)

ABSTRACT. The incidence of pupal diapause was examined in nine species of the genus Boettcherisca Rohdendorf ( B.peregrina, B.javanica, B.nathani, B.septentrionalis, B. karnyi, B. koimani, B. timorensis, B. formosensis and B.invaria ) collected from various localities in East Asia and Oceanian islands.
Colonies of B.septenrionalis, B. nathani and B.peregrina obtained from temperate regions of Asia and those of B. formosensis and B.peregrina from the continental subtropics entered pupal diapause in response to photoperiod at 20C.
The incidence of pupal diapause showed a geographic gradient in B.peregrina. No photoperiodically induced diapause was observed in the other five species which are indigenous to the tropical or subtropical habitats.
Different species of Boettcherisca seem to have evolved their pupal diapause independently of one another as an ecophysiological mechanism which enabled those species to expand from the tropical to temperate region.     

Cytogenetics of the genus Parasarcophaga (Sarcophagidae: Diptera)

Somatic chromosome complements of five sympatric species of the genus Parasarcophaga, viz. P. misera, P. albiceps, P. argyrostoma, P. ruficornis and P. knabi, are described. All the species have five pairs of meta/submetacentric autosomes and an XX/XY sex chromosome pair which is highly variable in size and shape. In P. misera and P. albiceps they are tiny dots while in P. knabi the metacentric X and Y chromosomes constitute almost one third of the genome. In P. ruficornis and P. argyrostoma they are telocentric chromosomes of moderate size. A comparative study of the C-banding patterns of P. ruficornis, P. knabi, P. argyrostoma and P. misera shows that autosomes of the former three species possess characteristic C-bands in pericentric regions while in P. misera they are absent. The heterochromatic sex chromosomes are C-band positive in all the species. However, with the exception of the tiny sex chromosomes of P. misera, the X chromosomes of other species show shorter or longer regions which stain rather lightly. These C-banded areas correspond to the heterochromatic areas revealed in orcein stained preparations. The evolutionary implications of these results are discussed.