重金属在拟水狼蛛体内的分布及对其体内抗氧化酶活性的影响

为了探明重金属在拟水狼蛛Pirata subpiraticus体内的分布及对其体内抗氧化酶活性的影响,本研究于2009年6月在河南南阳地区5种不同生境下,共采集50份土壤样本和300头拟水狼蛛样本,采用原子吸收光谱法测定了5种不同生境下雄雌拟水狼蛛体内重金属的分布、GSH含量以及GST,CAT和SOD的活性。结果表明：5个采集样点（S1, S2, S3, S4和S5） 拟水狼蛛体内重金属（Cd,Pb,Cu和Zn）的含量差异显著（P<0.05）。同一地点拟水狼蛛体内重金属（Cd,Pb,Cu和Zn）的含量不同身体部位差异显著（P<0.05）,头胸部>足部>腹部,同一地点雄性拟水狼蛛重金属含量显著高于雌性（P<0.05）。在重金属胁迫下,重金属含量高（S1,S2,S3和S4）的拟水狼蛛体内GSH 含量显著高于参照组（S5）,雄性GSH 含量高于雌性（P<0.05）。对于不同身体部位,GSH 含量差异显著,头胸部GSH 含量最高,其次为足部,腹部含量最低;GSH 含量与重金属含量显著正相关（r2=0.9854,P<0.05）。对于GST,CAT和SOD,重金属含量高则酶活性低,雄性酶活性显著低于雌性,不同身体部位酶活差异不显著;GST,CAT和SOD酶活性与重金属（Cd和Pb或者Pb）含量显著负相关。因此检测拟水狼蛛不同身体部位的酶活性变化就可知环境中重金属的污染程度,拟水狼蛛可以作为重金属污染的重要监测指示生物。     

Highly Fibrous and Porous Raw Material Shaped Chitin Isolated from <Emphasis Type="Italic">Oniscus</Emphasis> sp. (Crustacea)

Chitin was isolated from a crustacean by keeping the original body shape for the first time. The isolation method followed in this study was simple and time and energy saving unlike the labor-intensive classical methods. Chitin samples preserving the original shape were isolated from Isopoda (Oniscus sp.) successfully in a total of 20 min including filtration time. FT-IR, XRD and TGA analysis and chitinase digestion test demonstrated that the chitin was pure, low crystalline (Crystalline index: 51 %) and had low thermal stability (maximum degradation temperature: 328.8 °C). SEM analysis revealed the highly fibrous structure of chitin. The chitin content of the whole body was found significantly higher (29.6?±?4.2 %) than the earlier reports. Interaction of chitin isolates with Bovine Serum Albumin protein were studied at different pH. It was concluded that this three dimensional raw material shaped chitin can be effectively used in any adsorption studies due to 1) highly fibrous and porous nature, 2) low crystallinity and 3) low thermal stability. And also this biodegradable and biocompatible biopolymer can be suggested as a carrier matrix in further studies thanks to its three dimensional shape.     

Effects of carbon and nitrogen sources on the induction and repression of chitinase enzyme from<Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> isolates

Metarhizium anisopliae, an entomopathogenic hyphomycete, is being used effectively in Integrated Pest Management (IPM) system. Foliar application of these fungi is quite satisfactory as it invades its host by adhering to insect cuticles and formation of penetration structures called appresoria, which produces various extracellular enzymes, including chitinase that causes the insect cuticle breaching. The induction and repression mechanism of chitinase activity is not entirely understood and activity of this enzyme is different in response to different carbon and nitrogen sources. This report illustrates the effect of two carbon sources viz. colloidal chitin and dextrose and a nitrogen source, yeast extract on the chitinase production of fourteenM. Anisopliae isolates. The chitinase activity varied among the isolates and the different media used. A high enzymatic activity was observed in the medium containing an extra nitrogen source (yeast extract) followed by the medium containing colloidal chitin as a sole source of carbon and nitrogen. The exochitinase activity and the chitinase activity gel were also studied for the isolates showing high chitinase enzyme production. An array of chitinase isozymes were observed on chitinase activity gel with a common 14.3 kDa enzyme for all the isolates.     

雄性昆虫性信息素的研究进展

全面概述雄性昆虫性信息素的研究进展。主要从性信息素的释放机制及其作用、已发现的雄性昆虫性信息素及其化学结构进行阐述。通过不同部位释放的雄性昆虫性信息素种类是多样的,但其化学结构的差异又是细微的,雄性昆虫则利用这些差异来引诱雌性昆虫、促进交配和生殖隔离等。最后对雄性昆虫性信息素的研究作了展望。     

Applications of magnetic resonance spectroscopy to chitin from insect cuticles

Chitin is the second most abundant polysaccharide in nature after cellulose. At the present time, the main commercial sources of chitin are the crab and shrimp shells which are major waste products from the seafood industry. However, current chitin resources have some inherent problems including seasonal availability, limited supplies, and environmental pollution. As an alternative, insect cuticle is proposed as an unconventional but viable source of chitin. This review focuses on the recent sources of insect chitin and the application of various magnetic resonance spectroscopic techniques to native insect cuticles, particularly cicada sloughs and chitin extracted from insect sloughs. In addition, the physicochemical properties, isolation process, and degree of N-acetylation (DA) is reviewed and discussed.     

用裂解气液色谱法鉴定昆虫包涵体病毒的初步研究

用Shimadzu GC-9A气相色谱仪和PyR-2A管式炉裂解器,对11株昆虫包涵体病毒进行了裂解气相色谱鉴定,通过对“指纹图”的分析,既可明显地区分GV、NPV和CPV包涵体病毒彼此间的差异,亦可较好的区别不同分离株间的异同,实验结果初步证明,用裂解气液色谱法分析、鉴定昆虫包涵体病毒是可行的。     

The extracellular constitutive production of chitin deacetylase in Metarhizium anisopliae: possible edge to entomopathogenic fungi in the biological control of insect pests

The possible contribution of extracellular constitutively produced chitin deacetylase by Metarhizium anisopliae in the process of insect pathogenesis has been evaluated. Chitin deacetylase converts chitin, a beta-1,4-linked N-acetylglucosamine polymer, into its deacetylated form chitosan, a glucosamine polymer. When grown in a yeast extract-peptone medium, M. anisopliae constitutively produced the enzymes protease, lipase, and two chitin-metabolizing enzymes, viz. chitin deacetylase (CDA) and chitosanase. Chitinase activity was induced in chitin-containing medium. Staining of 7.5% native polyacrylamide gels at pH 8.9 revealed CDA activity in three bands. SDS-PAGE showed that the apparent molecular masses of the three isoforms were 70, 37, and 26 kDa, respectively. Solubilized melanin (10microg) inhibited chitinase activity, whereas CDA was unaffected. Following germination of M. anisopliae conidia on isolated Helicoverpa armigera, cuticle revealed the presence of chitosan by staining with 3-methyl-2-benzothiazoline hydrazone. Blue patches of chitosan were observed on cuticle, indicating conversion of chitin to chitosan. Hydrolysis of chitin with constitutively produced enzymes of M. anisopliae suggested that CDA along with chitosanase contributed significantly to chitin hydrolysis. Thus, chitin deacetylase was important in initiating pathogenesis of M. anisopliae softening the insect cuticle to aid mycelial penetration. Evaluation of CDA and chitinase activities in other isolates of Metarhizium showed that those strains had low chitinase activity but high CDA activity. Chemical assays of M. anisopliae cell wall composition revealed the presence of chitosan. CDA may have a dual role in modifying the insect cuticular chitin for easy penetration as well as for altering its own cell walls for defense from insect chitinase.     

Brazilian Angiostrongylus cantonensis haplotypes,ac8 and ac9, have two different biological and morphological profiles

Angiostrongylus cantonensis is the etiologic agent of eosinophilic meningoencephalitis in humans. Cases have been recorded in many parts of the world, including Brazil. The aim of this study was to compare the differences in the biology and morphology of two different Brazilian haplotypes of A. : ac8 and ac9. A significantly larger number of L1 larvae eliminated in the faeces of rodents at the beginning of the patent period was observed for ac9 haplotype and compared to the total of L1 larvae eliminated, there was a significant difference between the two haplotypes. The ac9 haplotype showed a significant difference in the proportion of female and male specimens (0.6:1), but the same was not observed for ac8 (1.2:1). The morphometric analysis showed that male and female specimens isolated from ac8 haplotype were significantly larger with respect to body length, oesophagus length, spicule length (male) and distance from the anus to the rear end (female) compared to specimens from ac9. The morphological analysis by light microscopy showed little variation in the level of bifurcations at the lateral rays in the right lobe of the copulatory bursa between the two haplotypes. The biological, morphological and morphometric variations observed between the two haplotypes agree with the observed variation at the molecular level using the cytochrome oxidase subunit I marker and reinforce the possible influence of geographical isolation on the development of these haplotypes.     

昆虫海藻糖酶的基因特性及功能研究进展

海藻糖酶(Treh)是昆虫能量代谢必不可少的一类酶, 亦是昆虫体内几丁质合成通路的第一个酶。其基因表达和酶活性直接与正常发育、 蜕皮、 变态以及繁殖等昆虫重要生理过程密切相关。目前已有多种昆虫的海藻糖酶基因被成功克隆, 从而发现昆虫海藻糖酶基因家族由多个成员组成。海藻糖酶基因所编码的蛋白大多数具有一个信号肽前导区, 部分蛋白拥有1~2个跨膜结构域, 根据是否具有跨膜结构, 可将其分为可溶性海藻糖酶(Treh1)和膜结合型海藻糖酶(Treh2)两类, 膜结合型海藻糖酶具有2个特有的标签序列, 即“PGGRFREFYYWDSY”和“QWDYPNAWPP”。海藻糖酶的主要功能是将胞外和胞内的海藻糖降解成葡萄糖, 为昆虫的生命活动提供能量。具体表现为两个方面, 一是参与昆虫几丁质合成途径, 从而调控表皮、 中肠等处的几丁质合成; 二是通过与激素的协同作用, 调控昆虫体内海藻糖和葡萄糖等糖类物质的浓度变化, 从而有效保护体内细胞的适应并渡过相应的逆境环境, 并提高其抗逆能力。鉴于海藻糖酶的重要功能, 其已成为害虫控制的潜在新靶标。不同类型海藻糖酶的功能研究及酶抑制剂的研发与应用将进一步推动害虫生物防治的发展。     

Visual Cues in Mate Recognition by Males of the Damselfly, Coenagrion puella (L.) (Odonata: Coenagrionidae)

Coenagrion puella males search actively for mates and are not aggressive to other males. To study the role of visual cues in male–female discrimination, four types of models were used: (1) bodies of intact insects, (2) models of painted males, (3) models of male–female chimerae, and (4) models of female body parts. Abdomen coloration pattern and presence of wings were the most important cues for sexual recognition by males. Step-by-step elimination of male coloration pattern leads to an increase in the tandem response rate. A female model painted as a male repelled males like the intact male model. The absence of either the head or the thorax slightly decreased the number of tandem responses, but models without both the head and the thorax were not recognized as a mate. Abdomen thickness larger than that of a normal female decreased the attractiveness of the model. Models of the gynochrome female were significantly more attractive than models of the androchrome one. Female models containing male parts were less attractive than models without any structure at this place. Using principal-components analysis, it is shown that models repelling males usually were those containing an intact male abdomen or a female abdomen painted with blue. The results indicate that C. puella males can distinguish males from females visually by morphological structures and coloration pattern.     

How Taxonomic Relations Affect the Physicochemical Properties of Chitin

Chitin specimens from 16 arthropod species (13 of Insecta and 3 of Arachnida) were isolated for the first time using the same method. Fourier Transform Infrared Spectrometry (FTIR), Thermogravimetric Analysis (TGA), X-ray diffraction (XRD), Scanning Electron Microscope (SEM) and elemental analysis have been applied to determine how physicochemical properties of chitin specimens are affected by taxonomic relationship. The characterisation studies revealed that physicochemical nature of the chitin specimens differed greatly and were found partially specific to taxa. Significant differences in the surface morphologies of chitin specimens were observed even in the same order. However, the chitin contents were recorded to be specific to the order in the class Insecta. The highest chitin content was observed in Coleoptera (18.2–25.2 %) followed by Hemiptera (10.6–14.5 %), Odonata (9.5–10.1 %), Hymenoptera (7.8–9.3 %), Diptera (8.1 %), Blattodea (4.7 %). In addition, the crystalline index (CrI) values of chitin specimens from Coleoptera were found to be higher than the other orders in Insecta. This study revealed that the chitin contents and CrI values can be related to taxonomical relationships.     

Fungi associated with the Egyptian cotton leafwormSpodoptera littoralis Boisdoval

A total of 28 species representing 15 fungal genera were found to be associated with the all different stages of the Egyptian cotton leafworm,Spodoptera littoralis Boisd., its faeces and the air of the containers where it was reared on Czapek Dox agar medium at 28°C.Aspergillus was the most common genus and was frequently isolated from all substrates.A. flavus was the only fungus associated with all insect stages. At least two well known facultative pathogens were obtained:Beauveria alba andCephaliophora tropica. Nearly all fungi recovered from insect faeces or its environment were also encountered from different insect stages. Forty-two isolates were tested for their abilities for growth on a synthetic medium (nitrogen- and carbon-free) containing partially purified chitin. Most (76.2%) had moderate growth rates.     

Seasonal variation of black fly (Diptera: Simuliidae) species diversity and community structure in tropical streams of Thailand

Seasonal variation of the physicochemical conditions of streams plays an important role in aquatic insect diversity and community structure. Asian tropical streams profoundly change between seasons due to the effects of monsoons. However, little is known about how these changes affect aquatic insect diversity and community structure. The objectives of this study are to examine seasonal variations of the physicochemical conditions in tropical streams in Thailand and to assess the effects of these changes on black fly community structure and diversity. Black flies were sampled and physicochemical conditions recorded at eight sites between December 2007 and December 2008. A total of 10 black fly species were found. Comparisons of the streams between seasons revealed that physical conditions related to rainfall rate were significantly different. Canonical correspondence analysis differentiated sampling sites from each season. Streams in the rainy season were faster and deeper, with higher discharge and conductivity than those of the cold and dry seasons. Species richness was significantly higher in the rainy season than in the cold and dry seasons ( F  = 6.23, P  = 0.004). Community structure profoundly changed between the low-flow season (cold and dry) and high-flow season. Black fly species found predominantly in the low-flow season ( Simulium siamense "cytoform A", S. aureohirtum ) decreased dramatically during the high-flow season. In contrast, species found at high frequency during the high-flow season ( S. nakhonense , S. angulistylum ) disappeared in the low-flow season. The study demonstrates the importance of seasonal variation of stream conditions on black fly community structure and diversity.     

Activity of glutathione S‐transferase and esterase enzymes in Helicoverpa armigera (Hübner) after exposure to entomopathogenic fungi

Helicoverpa armigera, a polyphagous insect of crops and vegetables, is acquiring resistance against many commercial insecticides. The present study shows variations in the activity of two detoxification enzymes, namely esterase and glutathione S‐transferase (GST), in H. armigera after exposure to different isolates of entomopathogenic fungi. After treatment of larvae with the different isolates (Day 0), samples were collected on three days (Days 3, 5 and 7) for enzyme analysis. High GST activity was found in samples of hemolymph, intestine and fat bodies of H. armigera following treatment with Beauveria bassiana (isolate Bb‐08), Metarhizium anisopliae (isolates Ma‐11.1 and Ma‐4.1), and Isaria fumosorosea (isolates If‐02 and If‐2.3). High esterase activity was recorded in samples of the intestine and fat bodies on various days after treatment, whereas increased esterase activity in hemolymph was noted only in samples from Day 5 after treatment with M. anisopliae (Ma‐4.1). The detection of high GST and esterase activity demonstrates the possibility of the development of resistance against these microbial control agents in H. armigera.     

Evaluation of the RAPD profiles from different body parts of Euglossa pleosticta Dressler male bees (Hymenoptera: Apidae, Euglossina)

In the current literature, information is scarce on which part of the adult insect body is suitable for isolation of genomic DNA for genetic analysis based on DNA-markers. In this study, we evaluated RAPD profiles generated from total genomic DNA isolated from distinct body parts (head, legs, thorax + wings and abdomen) of 12 males of Euglossa pleosticta Dressler. From the total of bands analyzed, 9.0% did not show reproducibility. Percent variations of bands in each body segment were: 1.1% (head); 0.4% (legs); 0.8% (thorax/wings) and 6.7% (abdomen). The much higher variation (chi2(one sample) = 10.27; df = 1; P < 0.01) in the RAPD profiles obtained by using DNA isolated from abdomen of the euglossine males suggests that this body part of adult insects should be avoided in DNA extraction procedures. Conversely, the low variation among the RAPD profiles obtained from amplifications of genomic DNA extracted from head, legs and thorax/wings indicates that all these body parts of male bees are equally useful and secure for using in isolation and amplification procedures of total genomic DNA.     

蛾类昆虫雄性信息素及其功能

昆虫性信息素是两性通讯系统的基础,其中雄性信息素的研究相对较少。本文综述了蛾类昆虫雄性信息素的研究进展。迄今已鉴定出40余种蛾类昆虫的雄性信息素,其行为学功能主要有对雌性的引诱和激欲、对同种雄性的抑制及种间隔离等。     

大气氟污染源附近食桑昆虫中氟的积累和分布

对大气F污染源附近野桑蚕,桑赤蠖和桑蚕体内的氟化物含量和器官分布进行了研究,结果表明,污染源附近食桑昆虫体有较高的氟化物含量,且食桑昆虫体的氟化物含量随着离污染源的距离增大而降低,野桑蚕,桑赤蠖和桑蚕的氟化物含量与桑叶的氟化物含量有极显著的线性正相关性,食桑昆虫不同器官间的氟化物积累量也存在较大差异,其消化管是食桑昆虫的主要氟化物积累器官。     

Effect of Toxic Heavy Metal Containing Industrial Effluent on Selected Life History Traits,Adult Morphology and Global Protein Expression Pattern of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

Accumulation of heavy metal laden industrial effluent into natural aquatic body leads to disruption of balance of that ecosystem as well as imperils the life cycle of the organisms living thereof. The present work aims to demonstrate the effect of toxic heavy metal containing industrial effluent on several life history traits, adult morphology and global protein expression pattern of insect model Drosophila melanogaster. Flies were reared for five generations in the effluent containing medium. One pair of virgin male and female fly from the stock was transferred to the vials. On the next day the male was removed and the female was transferred to fresh vials for successive days. Daily egg production of the females in different trials was recorded. Developmental time, pupation height, sex-ratio, egg-to-adult viability and mean body and wing length of the emerging flies from both treated and control groups were measured. We noted a drop in mean daily egg production rate, reduction in developmental time, shortening of pupation height, decreased egg-to-adult viability and a reduction in body and wing length in treated group in comparison to the control. However, no significant change was found in the sex-ratio of treated adult flies. Alterations in the expression pattern of body proteins in all the developmental stages were revealed by SDS-PAGE.     

Morphology of the microtubule system during the sphering, layering and polarization of normal and transformed fibroblasts]

The ultrastructure of the genital system (ductus seminalis, vestibulum, ductus tortuosus and canaliculus fecundans) has been studied during mating and oviposition. All these organs have a common organization and contain three parts: muscular and epithelial layer and chitin intima lining the interior surface of the female genital system. Ultrastructures of the muscular tissue and of the basal part of epidermis are similar. The luminal surface and microvilli are different in various organs of the female genital system. The ultrastructural pattern and the thickness of chitin intima may also vary, and can be used as a character for identificating different organs of the female system in ultrathin sections.