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Inflammatory bowel diseases (IBD) are a group of chronic disorders occurring in the intestinal tract. Previous studies demonstrated that genetics and microbiota play critical roles in the pathogenesis of IBD. Discoveries of genes that may regulate the homeostasis of gut microbiota and pathogenesis of IBD have the potential to provide new therapeutic targets for IBD treatment. The results suggested that the expression level of microRNA (miR)-602 is negatively related to the development of IBD, and that miR-602 overexpression in mice may prevent inflammation and intestinal barrier injuries in dextran sulfate sodium (DSS)-induced IBD mice. It was also found that the microbiota is important for miR-602-mediated prevention of IBD, as the inhibitory effect of miR-602 was lost when the microbiota was depleted using antibiotics. Furthermore, co-housing or adoptive transfer of microbiota from miR-602 could attenuate the pathogenesis of IBD. In addition, it was demonstrated that miR-602 could target tumor necrosis factor receptor-associated factor 6 (TRAF6) in intestinal epithelial cells. Collectively, the present results suggest that miR-602 plays a protective role in DSS-induced IBD by targeting TRAF6 in a microbiota-dependent manner.

Little is known about the firm-level dynamics behind trade responses to political tensions. This article reinvestigates variation in the travel pattern of the 14th Dalai Lama to study how political tensions affect trading decisions of Chinese importers. Using monthly trade data from China Customs covering imports of machinery and transport equipment from 173 countries over the 2000–2006 period, our empirical results show a significant reduction of imports in response to foreign government members' meetings with the Dalai Lama. In line with the idea that Chinese importers face a trade-off between bearing costs from suboptimal trade transactions and costs from not accommodating the government, this ‘Dalai Lama Effect’ operates at the intensive margin, i.e., via a decrease in the import volume per importer. Examining differential effects across types of firm ownership, we find that the observed effect is driven by state-owned enterprises (and foreign-invested firms) and not by private companies. Moreover, while direct importers temporarily reduce their trade with Dalai Lama-receiving countries, there is some evidence that trade intermediaries fill the void. Overall, we find the effects to be much more short-lived than previously thought.

A series of podophyllotoxin derivatives (M1–M16) that were selectively acylated by various phenoxy acids at the C-4 position of podophyllotoxin were synthesized, and their biological activities were also evaluated. Among them, compound M4 showed the most potent anti-cancer activity against HeLa cells with an IC50 value of 1.64 ± 0.41 μM. Additionally, flow cytometry analysis results indicated that it could cause a remarkable cell cycle arrest at G2/M phase, but the effect on apoptosis inducing was not significant. Moreover, the expression of cell cycle relative protein CDK1 was up regulated while cyclin B1 and Cdc25C, two proteins required for mitotic initiation were down regulated. Furthermore, the confocal assay and extracellular polymerized tubulin expression analysis also demonstrated that M4 was a potent tubulin polymerization inhibitor and the effect was comparable to that of colchicine. Finally, docking simulation results showed that M4 could nicely bind to the colchicine binding site of tubulin which further comfirmed the tubulin inhibiton activity of M4.

Numerous Bt rice lines expressing Cry protein derived from Bacillus thuringiensis Berliner (Bt) have been developed since 1989. However, the potential risks posed by Bt rice on non-target organisms still remain debate. The white-backed planthopper (WBPH), Sogatella furcifera (Horvath), is one of the most economically important insect pests of rice in Asian countries and also one of the main non-target herbivores of transgenic rice. In the current study, impacts of transgenic cry1Ab/vip3H+epsps rice (G6H1) with both insect and herbicide resistance on WBPH were evaluated to ascertain whether this transgenic rice line had potential risks for this sap-sucking pest under laboratory and field conditions. The laboratory results showed that no significant difference in egg developmental duration, nymphal survival rate and female fecundity was found for WBPH between G6H1 and its non-transgenic isoline (XS110). However, the development duration of nymphs was significantly shorter and female longevity significantly longer when WBPH fed on G6H1 by comparison with those on its control. To verify the results found in laboratory, a 3-yr field trial was conducted to monitor WBPH population using both the vacuum-suction machine and beat plate methods. Although the seasonal density of WBPH nymphs and total density of nymphs and adults were not significantly affected by transgenic rice regardless of the sampling methods, the seasonal density of WBPH adults in transgenic rice plots was slightly lower than that in the control when using the vacuum-suction machine. Based on these results both from laboratory and field, it is clear that our tested transgenic rice line will not lead higher population of WBPH. However, long-term field experiments to monitor the population dynamics of WPBH at large scale need to be conducted to confirm the present conclusions in future.

A 2-yr field trial was conducted to assess the impacts of two new transgenic Bt rice lines, T1C-19 expressing Cry1C protein and T2A-1 expressing Cry2A protein, on the arthropod community sampled via vacuum. All the arthropods were classified into five guilds, including herbivores, parasitoids, predators, detritivores, and others. The seasonal density and dominance distribution of each guild and community-level indices (species richness, Shannon-Wiener diversity index, Simpson diversity index, and evenness index) were compared among rice types. Principal response curves were used to investigate the differences of entire arthropod community of Bt rice plots relative to non-Bt rice plots. The results showed no significant difference was detected in the community-level indices and dominance distribution of guilds between Bt and non-Bt rice plots. The seasonal density of herbivores, detritivores, and others as well as density of the arthropod overall community were also not significantly affected by rice types in either year, although the density of predators and parasitoids in Bt rice plots was significantly lower than those in non-Bt rice plots. The lower abundances of Braconidae, Eulophidae, Cyrtorhinus lividipennis (Reuter) (Hemiptera: Miridae), and Theridiidae in Bt rice plots are likely attributed to the lower abundances of prey species or hosts. Principal response curves revealed that arthropod community in Bt was similar with that in non-Bt rice plots. In conclusion, our findings indicate that these two tested Bt rice lines had no marked negative effects on the arthropod community in the paddy fields.

Natural populations of Boettcherisca (Sarcophaga) peregrina Robineau-Desvoidy (Diptera: Sarcophagidae) were maintained for 20 generations and reared either on unpolluted diet or on polluted diet containing copper at a median lethal concentration (LC50) determined every five generations. This resulted in two reliable strains: the relative susceptible strain (S) and the copper-resistant strain (R). The metal accumulation, growth and development, reproduction, and antioxidant enzymes were analyzed in the two strains. The results showed that compared with the S strain, the R strain showed increased metal accumulation and fecundity of female adults. Regardless of whether larvae were fed on diet with or without Cu2+, the R strain showed higher activity of superoxide dismutase and glutathione S-transferase than the S strain, although without statistical significance. Moreover, the activity of superoxide dismutase and glutathione S-transferase increased when B. peregrina larvae were exposed to Cu2+ at 100 µg/g but decreased when they were exposed to Cu2+ at 800 µg/g. Larval catalase activity in the R strain was higher than in the S strain when larvae were fed on diet with or without Cu2+, although these differences were significant only at the 100 µg/g concentration. Moreover, the activity of catalase decreased when larvae were exposed to experimental Cu2+. Beyond all expectations, larval glutathione reductase activity was not significantly different between the two strains but changed slightly when larvae were exposed to experimental Cu2+. These results indicate that copper resistance in B. peregrina larvae is mediated by superoxide dismutase, catalase, and glutathione S-transferase. These results also help in establishing a physiological link between antioxidase activity and the resistance level of B. peregrina to copper.

BACKGROUND Transgenic rice producing the insecticidal protein from Bacillus thuringiensis Berliner (Bt) is protected from damage by lepidopteran insect pests. However, one of the main concerns about Bt rice is the potential impact on non-target herbivores. In the present study, the ecological impacts of two Bt rice lines, T1C-19 expressing Cry1C protein and T2A-1 expressing Cry2A protein, on the non-target herbivore brown planthopper (BPH), Nilaparvata lugens (Stal), were evaluated under laboratory and field conditions. The purpose was to verify whether these Bt rice lines could affect the performance of BPH at individual and population scales. RESULTS Laboratory results showed that most of the fitness parameters (development duration, survival rate, fecundity, fertility, amount of honeydew excreted) of BPH were not significantly affected by the two tested Bt rice lines, although the development duration of fourth-instar nymphs fed on T1C-19 was distinctly longer compared with that on T2A-1 and non-Bt rice plants. Five life-table parameters did not significantly differ among rice types. Two-year field trials also revealed no significant difference in population dynamics of BPH among rice types. CONCLUSION It is inferred that the tested Bt rice lines are unlikely to affect the population growth of BPH. © 2014 Society of Chemical Industry

Transgenic rice expressing Bacillus thuringiensis Berliner (Bt) protein can effectively control target insects including stem borers and leaf folders. However, the potential effects of Bt rice on nontarget organisms including nontarget herbivores have not been fully evaluated. In the current study, ecological fitness parameters of the nontarget herbivore, Nephotettix cincticeps (Uhler) (Hemiptera: Cicadellidae), fed on T1C-19 (Cry1C) or T2A-1 (Cry2A) rice were compared with non-Bt rice (MH63) under laboratory conditions. A 2-yr field trial was also conducted to monitor the population dynamics of N. cincticeps in the Bt and control rice plots using the vacuum-suction machine and yellow sticky card traps. Laboratory results showed that there were no significant differences in some of biological parameters including egg developmental duration, adult fresh weight, adult longevity, and oviposition period when N. cincticeps fed on Bt or non-Bt rice was compared. However, the survival rate of N. cincticeps nymphs fed on T2A-1 Bt rice plants was significantly higher than that on the control. When N. cincticeps fed on T1C-19 Bt rice plants, its nymphal duration was significantly longer and fecundity significantly lower compared with those fed on both T2A-1 Bt and non-Bt rice plants; the preoviposition period of N. cincticeps fed on T1C-19 and T2A-1 Bt rice was also significantly shorter than those on non-Bt rice. Nonetheless, both seasonal density and population dynamics of N. cincticeps adults and nymphs were similar between Bt (T1C-19 and T2A-1) and non-Bt rice plots under field conditions. In conclusion, our results indicate that our two tested Bt rice lines would not lead to higher population of N. cincticeps. Long-term experiments to monitor the population dynamics of N. cincticeps at large scale need to be carried out to confirm the current results.

Tea is one of the most economically important crops in China. To secure its production and quality, biological control measures within the context of integrated pest management (IPM) has been widely popularized in China. IMP programs also provide better control of arthropod pests on tea with less chemical insecticide usage and minimal impact on the environment. More than 1100 species of natural enemies including about 80 species of viruses, 40 species of fungi, 240 species of parasitoids and 600 species of predators, as well as several species of bacteria have been recorded in tea ecosystems in China. Biological and ecological characteristics of some dominant natural enemies have been well documented. Several viral, bacterial, and fungal insecticides have been commercially utilized at large scale in China. Progress in biological control methods in conjunction with other pest control approaches for tea insect pest management is reviewed in this article. Knowledge gaps and future directions for tea pest management are also discussed.

As a key component of ecological risk assessments, nontarget effects of Bacillus thuringiensis (Bt) rice have been tested under laboratory and field conditions for various organisms. A 2-yr field experiment was conducted to observe the nontarget effects of six transgenic rice lines (expressing the Cry1Ab or fused protein of Cry1Ab and Cry1Ac) on four nontarget thrips species including Frankliniella intonsa (Trybom), F. tenuicornis (Uzel), Haplothrips aculeatus (F.), and H. tritici (Kurd), as compared with their rice parental control lines. Two sampling methods including the beat plate and plastic bag method were used to monitor the population densities of the four thrips species for 2 yr. The results showed that the seasonal average densities of four tested thrips species in Bt rice plots were significantly lower than or very similar to those in the non-Bt rice plots depending on rice genotypes, sampling methods, and years. Among all six tested Bt rice lines, transgenic B1 and KMD2 lines suppressed the population of these tested thrips species the most. Our results indicate that the tested Bt rice lines are unlikely to result in high population pressure of thrips species in comparison with non-Bt rice. In some cases, Bt rice lines could significantly suppress thrips populations in the rice ecosystem. In addition, compatibility of Bt rice, with rice host plant resistance to nontarget sucking pests is also discussed within an overall integrated pest management program for rice.

Most molecular work on the roles of heat shock proteins (hsps) in host-parasite interaction has focused on vertebrates, rather than invertebrates. Here the full length complementary DNA (cDNA) sequences of three hsp genes (hsp20, hsp75 and hsp90) were amplified from Pieris rapae, and their transcriptional responsiveness to parasitization by the endoparasitic wasp Pteromalus puparum were investigated. The cDNA sequence analysis of hsp20, hsp75 and hsp90 revealed open reading frames of 531, 2 328 and 2 157 bp in length, which encode proteins with calculated molecular weights of 19.5, 75.48 and 82.7 kDa, respectively. The comparison of amino acid sequences showed that P. rapae hsp20 shared highly divergent homology to that of other insects, while hsp75 and hsp90 showed high homology to their counterparts of other species. The expression analysis indicated that these three genes were influenced in response to parasitization by P. puparum. The hsp20 transcripts in parasitized pupae were higher compared to non-parasitized pupae. The expression of hsp75 and hsp90 were down-regulated following parasitization. The results indicate that hsps are involved in host-parasitoid interactions.

Six transgenic rice, Oryza sativa L., lines (G6H1, G6H2, G6H3, G6H4, G6H5, and G6H6) expressing a fused Cry1Ab/Vip3H protein, were evaluated for resistance against the Asiatic rice borer, Chilo suppressalis (Walker) (Lepidoptera: Crambidae), and the stem borer Sesamia inferens (Walker) (Lepidoptera: Noctuidae) in the laboratory and field. The bioassay results indicated that the mortality of Asiatic rice borer and S. inferens neonate larvae on six transgenic lines from seedling to filling stage was up to 100% at 168 h after infestation. The cumulative feeding area by Asiatic rice borer neonate larvae on all transgenic lines was significantly reduced compared with the untransformed parental 'Xiushui 110' rice. A 2-yr field evaluation showed that damage during the vegetative stage (deadheart) or during the reproductive stage (whitehead) caused by Asiatic rice borer and S. inferens for transgenic lines was much lower than the control. For three lines (G6H1, G6H2, and G6H6), no damage was found during the entire growing period. Estimation of fused Cry1Ab/Vip3H protein concentrations using PathoScreen kit for Bt-Cry1Ab/1Ac protein indicated that the expression levels of Cry1Ab protein both in main stems (within the average range of 0.006-0.073% of total soluble protein) and their flag leaves (within the average range of 0.001-0.038% of total soluble protein) were significantly different among six transgenic lines at different developmental stages. Both laboratory and field researches suggested that the transgenic rice lines have considerable potential for protecting rice from attack by both stem borers.

Nontarget impacts of six transgenic Bt rice lines (expressing the Cry1Ab or Cry1Ab/Cry1Ac protein) on the thrips, Stenchaetothrips biformis (Bagnall), attacking the rice seedling and tillering stages, were evaluated under laboratory and field conditions. Laboratory results showed relatively longer larval, pupal development and preoviposition durations of S. biformis. Although it had a shorter oviposition period, female adult longevity and less total laid eggs were found when fed on some tested Bt rice in comparison to non-Bt controls. S. biformis population dynamics in Bt and non-Bt plots were monitored using the plastic bag and beat plate methods. In the field, the temporal patterns of S. biformis population changes were similar between tested Bt rice lines and their respective control; however, the total number of S. biformis individuals collected from the Bt plots were significantly less or the same, varying from variety to variety, compared with those from the non-Bt plots. ELISA results showed that the Bt insecticidal protein could be transferred from Bt rice to the thrips, and the concentrations of the protein in rice leaves and thrips were not significantly correlated with some important biological parameters of the thrip. In addition, the potential effects of Bt rice on the abundance of S. biformis candidate predators are also discussed. In conclusion, our results show that the six Bt rice lines assessed may be less preferable host plants to S. biformis at the individual and population levels in comparison to the non-Bt rice plants.

We screened an endoparasitic wasp (Pteromalus puparum) cDNA library for DNA sequences having antimicrobial activity using a vital dye exclusion assay. Two dozens of clones were isolated that inhibited the growth of host Escherichia coli cells due to expression of the cloned genes. Three peptides (PP13, PP102 and PP113) were synthesized chemically based on the amino acid sequences deduced from these clones and assayed for their antimicrobial activity. These peptides have net positive charges and are active against both Gram-negative and -positive bacteria, but are not active against fungi tested. Their hemolytic activity on human red blood cells was measured, and no hemolytic activity was observed after 1-h incubation at a concentration of 62.5 microM or below. A Blast search indicated that the three peptides have not been previously characterized as antimicrobial peptides (AMPs). Salt-dependency studies revealed that the biocidal activity of these peptides against E. coli decreased with increasing concentration of NaCl. Transmission electron microscopic (TEM) examination of PP13-treated E. coli cells showed extensive damage of cell membranes. The CD spectroscopy studies noted that the enhanced alpha-helical characteristics of PP13 strongly contribute to its higher antimicrobial properties. These results demonstrate the feasibility to identify novel AMPs by screening the expressional cDNA library.

Although the biochemical dissection of parasitoid-host interactions is becoming well characterized, the molecular knowledge concerning them is minimal. In order to understand the molecular bases of the host immune response to parasitoid attack, we explored the response of Papilio xuthus parasitized by the endoparasitic wasp Pteromalus puparum using proteomic approach. By examining the differential expression of plasma proteins in the parasitized and unparasitized host pupae by two-dimensional (2D) electrophoresis, 16 proteins were found to vary in relation to parasitization compared with unparasitized control samples. All of them were submitted to identification by mass spectrometry coupled with a database search. The modulated proteins were found to fall into the following functional groups: humoral or cellular immunity, detoxification, energy metabolism, and others. This study contributes insights into the molecular mechanism of the relationships between parasitoids and their host insects.

Pteromalus puparum is a predominant endoparasitoid wasp of Pieris rapae. Its venom is the only active factor injected into host associated with oviposition. In this report, we explored whether the venom alone from this wasp affects the endocrine system of its host or not. We monitored the changes of hemolymph juvenile hormone (JH; only JH III detected), ecdysteroid, and juvenile hormone esterase activity (JHE) over 72 h in parasitized and venom-microinjected P. rapae pupae. Non-parasitized and PBS-microinjected P. rapae served as controls. Results showed that JH titers were significantly higher in parasitized and venom-microinjected pupae than that in control pupae during 24 to 72 h. After 12 h, JH titers were significantly promoted by parasitization and venom microinjection. JHE activities of non-parasitized and PBS-microinjected pupae were significantly higher than that of parasitized and venom-microinjected pupae, which was with a peak at 12 h (parasitized pupae) or 24 h (venom-microinjected pupae) during 6 to 48 and 12 to 36 h, respectively. The hemolymph titers of ecdysteroid in non-parasitized and PBS-microinjected pupae increased rapidly during 12 to 36 h with a peak at 36 h, and were higher than treatments before 48 h, while presenting a significant difference at 24 to 48 h between the treatments and controls. The results demonstrate that venom alone of this parasitoid wasp can disrupt its host's endocrine system. © 2009 Wiley Periodicals, Inc.

Effects of copper (Cu) accumulation by the flesh fly Boettcherisca peregrina (R.- D.) (Diptera: Sarcophagidae) on the ectoparasitic wasp Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) were investigated experimentally by exposing host larvae to contaminated diets with final Cu concentrations of 400 � g/g and 800 � g/g diet fresh weight (DFW), respectively. Results showed that Cu can be transferred along food chains to secondary consumers (parasitoids) in small amounts, resulting in negative effects on parasitoid growth and development (body weight and developmental duration) as well as fecundity (number of offspring per female). Copper exposure also inhibited vitellogenesis of parasitoids from Cu-contaminated host pupae. It is suggested that the decreased fecundity and inhibition of vitellogenesis of N. vitripennis resulted from poor host nutritional state rather than from direct effects of Cu stress.

In hymenopteran parasitoids devoid of symbiotic viruses, venom proteins appear to play a major role in host immune suppression and host regulation. Not much is known about the active components of venom proteins in these parasitoids, especially those that have the functions involved in the suppression of host cellular immunity. Here, we report the isolation and characterization of a venom protein Vn.11 with 24.1 kDa in size from Pteromalus puparum , a pupa-specific endoparasitoid of Pieris rapae . The Vn.11 venom protein is isolated with the combination of ammonium sulfate precipitation and anion exchange chromatography, and its purity is verified using SDS–PAGE analysis. Like crude venom, the Vn.11 venom protein significantly inhibits the spreading behavior and encapsulation ability of host hemocytes in vitro . It is suggested that this protein is an actual component of P. puparum crude venom as host cellular-immune suppressive factor.

The relative toxicity of Bt rice pollen to domestic silkworm, Bombyx mori Linnaeus (Lepidoptera: Bombycidae), was assessed by a leaf-dip bioassay under laboratory conditions. Silkworm first instars were sensitive to pollen from Bt rice lines, B1 and KMD1, but were not sensitive to pollen from Bt rice line TT9–3. First instars were 1.34–2.12 times more sensitive to B1 pollen than older instars. Bioassays of subacute toxicity under a worst-case scenario suggested that continuous exposure to a sublethal dose of B1 pollen or equivalent doses of non-Bt rice pollen affected silkworm survival and development. Young larvae were more affected by continuous exposure to Bt pollen than older larvae but less affected by non-Bt pollen. Ultrastructural observations showed that Cry proteins associated with Bt pollen were released into the larval lumen and resulted in pathological midgut changes and negative impacts on silkworm survival and development. However, considering that the sublethal dose of Bt pollen (L...

Transgenic rice containing insecticidal protein(s) from Bt (Bacillus thuringiensis) effectively control lepidopteran pests under laboratory and field conditions. However, Bt rice has not been widely commercialized. A major concern regarding the deployment of Bt rice is its potential impact on non-target arthropods, including herbivorous insect pests and biological control agents (predators and parasitoids) in the rice ecosystem. A 2-year field study was conducted at 3 sites in Zhejiang Province, China to assess the impacts of a homozygous transgenic japonica rice line KMD1 (Bt rice) containing a synthetic cry1Ab gene from Bacillus thuringiensis Berliner. The impacts on three planthoppers (Homoptera: Delphacidae), Sogatella furcifera (Horvath), Nilaparvata lugens (Stal) and Laodelphax striatellus (Fallen), and the natural enemy, Cyrtorhinus lividipennis (Reuter) (Hemiptera: Miridae), were evaluated using two sampling methods: vacuum-suction and Malaise trap. Population densities of planthoppers and C. lividipennis were not significantly affected by rice type (Bt/non-Bt) over the season at any of the 3 sites in each year, regardless of sampling methods. Both in Bt rice and non-Bt rice plots, S. furcifera was the predominant species of planthoppers as determined by either of the two sampling methods, and comprised >50% of the planthoppers at each site. No consistent effects of Bt rice and Bt rice × sampling date interaction on population dynamics of the predominant planthopper species, S. furcifera, and the predator, C. lividipennis, were observed throughout the sampling period. Overall, this 2-year field study indicates that, in comparison with non-Bt rice, Bt rice did not lead to higher planthopper populations and did not negatively affect the predator C. lividipennis. These results suggest that use of Bt rice may provide control of the key lepidopteran pests while promoting the key biological control agent of planthopppers, C. lividipennis.

Age determination is the basis of determining the postmortem interval using necrophagous fly larvae. To explore the potential of using cuticular hydrocarbons for determining the ages of fly larvae, changes of cuticular hydrocarbons in developing larvae of Chrysomya rufifacies (Macquart) (Diptera: Calliphoridae) were investigated using gas chromatography with flame-ionization detection and gas chromatography-mass spectrometry. This study showed that the larvae produced cuticular hydrocarbons typical of insects. Most of the hydrocarbons identified were alkanes with the carbon chain length of 21–31, plus six kinds of alkenes. The hydrocarbon composition of the larvae correlated with age. The statistical results showed that simple peak ratios of n-C29 divided by another eight selected peaks increased significantly with age; their relationships with age could be modelled using exponential or power functions with R2 close to or > 0.80. These results suggest that cuticular hydrocarbon composition is a useful indicator for determining the age of larval C. rufifacies, especially for post-feeding larvae, which are difficult to differentiate by morphology.

In 2003 and 2004, field studies were conducted at three sites in Zhejiang Province in China to assess the impacts of Bacillus thuringiensis (Bt) Berliner rice expressing a fused gene of cry1Ab and cry1Ac on nontarget planthoppers and leafhoppers. Populations in Bt plots were sampled with yellow sticky card traps, Malaise traps, and a vacuum-suction machine, and compared with samples from non-Bt control (IR72) plots. The results from yellow sticky card trap samplings indicated no significant differences between Bt and non-Bt plots in the species composition or densities of each species of planthopper and leafhopper. Three species of planthoppers, Sogatella furcifera (Horvath), Nilaparvata lugens (Stal), and Laodelphax striatellus (Fallen), were collected at all sites, and three species of leafhoppers, Nephotettix cincticeps (Uhler), Thaia subrufa (Motschulsky), and Recilia dorsalis (Motschulsky), were collected at Hangzhou. Another species of leafhopper, N. virescens (Distant), was collected at Anji and Jiande instead of T. subrufa. The results from the Malaise trap and vacuum-suction samples revealed no significant differences between Bt and non-Bt plots in species structure of planthoppers and leafhoppers or in population changes of the predominant planthopper species, S. furcifera, the predominant leafhopper species, N. cincticeps, or N. virescens throughout most sampling dates. Densities of planthoppers and leafhoppers were significantly affected by year and site but not by Bt rice. In general, our results suggest that the Bt rice line tested did not adversely affect nontarget planthopper and leafhopper populations and will not lead to higher populations or damage by planthoppers and leafhoppers.

Some cry genes from Bacillus thuringiensis Berliner (Bt) have been transferred into rice plants for resistance to lepidopterans. The ecological risks of Bt rice, especially the non-target effects of Bt rice pollen on the domestic silkworm, Bombyx mori Linnaeus in mulberry-rice mixed cropping area should be clarified cautiously. In light of B. mori fully domesticated and not surviving independently in the field, a series of laboratory bioassays were conducted to evaluate the effect of the pollen from an indica transgenic rice line, TT9-3 with a fused cry1Ab/cry1Ac gene on B. mori based on the investigation of rice pollen deposition under normal field conditions and the quantitation of the fused insecticidal protein expression in the pollen. No significant adverse effects were observed on the survival, growth and development of B. mori young larvae, even after the neonates had been exposed to Bt pollen at the highest density of 3,395.0 grains/cm2 for 48 h which the pollen density is far higher than the highest pollen density on mulberry leaf, 1,635.9 grains/cm2, naturally occurred in the field. According to these results and considering the extensive biotic and non-biotic factors such as the type of Bt genes used and expressed in Bt rice line TT9-3 which are safe to B. mori, the relatively low exposure amount of insecticidal protein in the Bt pollen, and other conditions which influence the silkworm's exposure to Bt pollens, it suggests that the pollen from Bt rice line, TT9-3 poses little effect on silkworm rearing in natural settings.

Crude venoms from two parasitoid species, Pteromalus puparum and Nasonia vitripennis (Hymenoptera: Pteromalidae) were assayed for biological activities toward hemocytes from two species of their natural hosts and eight species of their non-natural hosts as well as two lines of cultured Lepidoptera cells, respectively. By inhibiting the spreading and viability of insect hemocytes, the venom from P. puparum displayed significantly higher activities toward plasmatocytes and granular cells from both larvae and pupae of two natural hosts, Pieris rapae and Papilio xuthus, and lower activity toward those from Spodoptera litura, Musca domestica and Sarcophaga peregrina. However, no effect was found towards any type of hemocytes from other five insects tested, namely, Ectropis oblique, Galleria mellonella, Sesamia inferens, Bombyx mori and Parnara guttata. In contrast, the venom from N. vitripennis showed a narrower range of targeted insects. It appeared to have highly adverse effects on the spreading and viability of plasmatocytes and granular cells only from the natural hosts, M. domestica and S. peregrina, little toxicity to cells from P. rapae and P. xuthus, and no effect on any of the other insects tested. Pteromalus puparum venom also apparently presented a high ability to block the spreading of Tn-5B1-4 cells derived from Trichoplusia ni, and high cytotoxicity to the cells and Ha cells derived from Helicoverpa armigera. Nasonia vitripennis venom, however, only had a marked lethal effect to Ha cells. In addition, the possibility that the host range of a defined parasitoid could be assessed using our method of treating hemocytes from candidate insects with venom in vitro, and the potential of our venoms tested in the development of bio-insecticides, insect-resistant transgenic plants, are discussed.

The effect of nitrogen content in rice plants on the tolerance of brown planthopper (BPH), Nilaparvata lugens Stal to high temperature, starvation and insecticide, was studied in the laboratory at International Rice Research Institute (IRRI), Philippines. Survival of nymphs and adults, fecundity and egg hatchability were significantly increased by the increase of nitrogen content in host plants at 38. Moreover, the survival of nymphs, fecundity and egg hatchability were significantly higher in BPH populations on rice plants with a high nitrogen regimen than those on rice plants with a low nitrogen regimen. Meanwhile, the tolerance of female adults to starvation and nymphs to growth regulator buprofezin on rice plants with a high nitrogen regimen were slightly increased. This indicates that the tolerances of BPH to adverse environmental stresses were positively increased by the application of nitrogenous fertilizer. The outbreak potential of BPH induced by the excessive application of fertilizer in rice fields was also discussed.

Photoperiodic sensitivity for diapause induction of the green lacewing, Chrysoperla sinica (Tjeder) was examined at 22°C. The adult diapause of C. sinica was induced by short-day photoperiods, and the critical photoperiod for its induction was between 12.5L-11.5D and 13L-11D. Adults developed without diapause under long-day conditions, and entered diapause under short-day conditions. The adult stage was the uppermost sensitive stage for adult diapause induction, adults could go into diapause only when the emerging adults were under diapause-inducing short-day photoperiods. The short-day photoperiodic experience by transferring between 15L: 9D and 9L: 15D at preimaginal stages did not result in adult diapause under 15L: 9D photo regime, although some treatments extended the pre-oviposition period in adult stage. The results showed that the 3rd instar larvae and pre-pupae were more sensitive to the photoperiodic change from 15L: 9D to 9L: 15D photo regime than the other preimaginal stages.

In contrast to the situation with egg-larval and larval endoparasitic wasps, little is known about the effects of pupal endoparasitoids and their secretions on the hemocytes of their insect hosts. This study focuses on the pupal endoparasitoid, Pteromalus puparum, and its host, the small white butterfly, Pieris rapae. Parasitism by P. puparum, resulted in a significant increase in the total number of host hemocytes up to day five after parasitization. From day one to day four after parasitization, the percentage of plasmatocytes significantly decreased, and the proportion of granular cells increased. Moreover, from 12 h to day three after parasitization, hemocyte mortality in parasitized pupae was noticeably higher. When P. rapae pupae were parasitized by adult females of P. puparum irradiated by gamma-ray (pseudoparasitization), it was clear that the treated wasps could induce similar hemocyte changes. However, such phenomena did not occur in punctured host pupae (mimic-parasitization). After treatment with P. puparum venom, both the percentages of spreading plasmatocytes and encapsulated Sephadex G-25 beads were lessened significantly in vitro. Electron microscopy analysis and visualization of hemocyte F-actin with phalloidin-FITC showed that hemocytes treated with venom had a rounded configuration and neither spread nor extended pseudopods, while there was no marked alteration of hemocyte cytoskeletons after venom treatment. The results suggested that venom of P. puparum could actively suppress the hemocyte immune response of its host, but not by destroying the host hemocyte cytoskeleton.

Ecological fitness characters of the brown planthopper increased proportionally with increase in nitrogen content of rice plants they were bred on. The nymphs survived better and had shorter life spans. Females weighed heavier, laid more eggs and lived longer. In addition, egg hatchability was significantly increased. Number of insects and dry mass from one pair of parents were also significantly higher in high nitrogen plants. Relationships between each of these characters with nitrogen content fitted the linear models well. These characters from insects reared on four successive generations in low, medium and high nitrogen regimes were also investigated. There was little between-generation difference in most characters, except for the population and its dry mass produced by one pairs of insects. This implies that in areas where nitrogen-rich crops are abundant over long periods, ecological fitness of the brown planthoppers are potentially higher and if natural biological control is compromised, the risk of extensive outbreaks are higher. Implications to sudden stoppage of insecticide use in these areas are also discussed.

Two transgenic rice (Oryza sativa L.) lines, KMD1 and KMD2 at the R4 generation, transformed with a synthetic cry1Ab gene from Bacillus thuringiensis Berliner, were first evaluated for stem borer resistance in the field during the rice growing season of 1998 in two areas of Zhejiang Province, China. Both KMD1 and KMD2 were highly resistant to the stem borers Chilo suppressalis (Walker) and Scirpophaga incertulas (Walker), and were completely undamaged during the whole rice growing season. In contrast, damage to the plants of the untransformed parental control (Xiushui 11) was in the form of deadhearts or whiteheads. Under natural infestation by the C. suppressalis, the damage to control plants reached a peak of 88.7% of plants and 20.1% of tillers encountered with deadhearts. Under artificial and natural infestation of neonate striped stem borers at the vegetative stage and booting stage, 100% of plants and 25.6% of tillers, 78.9% of plants and 15.6% of productive tillers among artificially infested control plants were observed with the symptom of deadhearts and whiteheads, respectively. Damage to the control plants from artificial infestation by the S. incertulas reached a peak of 97.0% of plants and 22.9% of tillers damaged. The field research indicated that both KMD1 and KMD2 show great potential for protecting rice from attack by these two stem borers.

Continuous rearing of the brown planthopper, Nilaparvata lugens (Stal), was achieved on a chemically defined diet D97 developed in this laboratory. The performance of N. lugens reared on D-97 was much better than that on a previously reported artificial diet MMD-1, although it was inferior to the susceptible rice variety TN1. N. lugens can not finish a complete generation on MMD-1 due to poor fecundity and unhatchability of eggs, but it has been successfully reared for more than 6 generations on D-97. In comparison with TN1-reared N. lugens, D-97-reared insects had a longer nymphal period (more than 3 days longer for males and 5 days longer for females), lighter weight (over 25% lighter for males and 35% lighter for females) and lower fecundity (over 50% less). Of the six generations on D-97, the first two generations had a significantly higher rate of emergence and hatchability. The possible factors influencing rearing efficiency and the potential applications of this diet are also discussed.

To fully explore the resistance potential of transgenic rice produced by Agrobacterium-mediated transformation, an elite line KMD1 was assessed for its resistance to eight lepidopteran rice pest species. KMD1 contained a synthetic cry1Ab gene from Bacillus thuringiensis under the control of a maize ubiquitin promoter. It was derived from a commercial japonica Chinese rice variety Xiushui 11, and bred true for both agronomic traits and a cry1Ab gene when the bioassays were done in 1998 in the R5 generation. The eight lepidopteran pest species were: four Pyralidae species: Chilo suppressalis (striped stem borer, SSB), Scirpophaga incertulas (yellow stem borer, YSB), Cnaphalocrocis medinalis (leaf folder), Herpitogramma licarisalis; two Noctuidae: Sesamia inferens (pink stem borer, PSB) and Naranga anescens; one Stayridae: Mycalesis gotama; and one Hesperiidae, Parnara guttata. In laboratory bioassays, 100% mortality was observed in all insect species when their newly hatched or third-instar larvae were fed KMD1 leaf tissues, whereas only 9.65% of the neonates and none of the third-instar larvae died when fed the leaf tissues of non-transgenic control. Moreover, the leaf area of control tissues consumed in four days by stem borers was 20 to 40 times higher than that of KMD1 tissues, and the area of control tissues eaten by leaf-feeding species was 120 to 180 times greater than that of the transgenic tissues. Under natural infestation, no KMD1 plant was visibly damaged by the SSB, YSB and leaf folder in field evaluation. On the other hand, 80, 9.3 and 88.7% of control plants were injured by SSB, YSB, and leaf folder, respectively. These data disclosed that the transgenic line was highly resistant to a broad spectrum of lepidopteran insect species and could be useful in insect resistance breeding programs.

The effect of a high temperature, i. e. 32°C. on vitellogenesis of the Japanese oak silkworm, Antheraea yamamai (Lepidoptera: Saturniidae) was markedly significant. Its extent was dependent on the development stage of the silkworm exposed to 32°C. When exposed to 32 °C since the 1st day after cocooning, titres of both vitellogenin (Vg) and soluble proteins in the fat body and hemolymph of mature larvae were evidently lower than those at 26°C. When pupae were maintained at 32°C since the 1st day after pupation. the titres of Vg in the fat body showed no significant difference from those at 26°C, but those in the hemolymph and the titres of vitellin (Vt) in the ovary mostly were obviously lower in contrast to those at 26°C. While exposed to 32°C since the 6th day after pupation, at most instance the tires of Vg both in the fat body and hemolymph were not markedly different from those at 26°C, and those of Vt in the ovary were significantly higher than those at 26°C, In addition, the changes in the titres of soluble proteins in the fat body and hemolymph as well as the ovary were monitored when pupae were maintained at 32°C since the 1st or 6th day after pupation. It is recommended that both mature larvae and pupae at cocooning stage and earlier pupal stage should not be exposed to 32°C when the silkworm is reared for egg raising.

A study of Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae) midgut structures and contents was conducted using scanning and transmission electron microscopy. The larval midgut was enclosed by a peritrophic membrane that seemed to be composed of two layers. Numerous bacteria were found throughout the lumen of the midgut, and because the midgut does not open to the hindgut, we hypothesized that the bacteria may serve to decompose the residues occurring in the midgut lumen. Few yeast cells were present in the larval midgut. The visual observations suggest that the concentration of bacteria were much lower in adult midguts than in larval midguts. However, many yeast cells were observed in the lumen of the adult midgut. No obvious peritrophic membrane was observed in electron micrographs of the adult midgut compared with the larval midgut. The results suggest different modes of food residue disposal.

BACKGROUND: The commercial release of rice genetically engineered to express a Cry1Ab protein from Bacillus thuringiensis (Bt) for control of Lepidoptera in China is a subject of debate. One major point of the debate has focused on the ecological safety of Bt rice on nontarget organisms, especially predators and parasitoids that help control populations of insect pests. METHODOLOGY/PRINCIPAL FINDINGS: A tritrophic bioassay was conducted to evaluate the potential impact of Cry1Ab-expressing rice on fitness parameters of a predaceous ground spider (Pardosa pseudoannulata (Bösenberg et Strand)) that had fed on Bt rice-fed brown planthopper (Nilaparvata lugens (Stål)) nymphs. Survival, development time and fecundity of this spider were not different when they were fed with Bt rice-fed or non-Bt rice-fed prey. Furthermore, ELISA and PCR gut assays, as well as a functional response trial, indicated that predation by P. pseudoannulata was not significantly different in Bt rice or non-Bt rice fields. CONCLUSIONS/SIGNIFICANCE: The transgenic Cry1Ab rice lines tested in this study had no adverse effects on the survival, developmental time and fecundity of P. pseudoannulata in the laboratory or on predation under field conditions. This suggests that this important predator would not be harmed if transgenic Cry1Ab rice were commercialized.

The Japanese oak silkworm, Antheraea yamami, is a precious wild silkworm. Through comparing survival rate, stadium, bodyweight, nutritional indexes, coconing rate and cocoon quality of the larvae reared on Quercus fabri fresh leaves with those on Q. acutissima, it was concluded that Q. fabri is a better fodder plant for this silkworm than Q. acutissima in East China.

Two aquatic plants, Zizania latifolia and Sagittaria sagittifolia, were used to study the variation of physiological responses and the water qualities under mild, moderate and weighty eutrophic water in plastic pot scale experiments. The physiological characteristics of both plants were investigated, which included contents of leaf malondialdehyde (MDA) and proline, membrane permeability (MP), and activities of catalase (CAT) and peroxides (POD). The water qualities were analyzed correspondingly. The results showed remarkable effects of different degree of eutrophic water on physiological characteristics of the plants. Compared with mild eutrophic water, increasing degree of eutrophication significantly increased the contents of MDA and MP, decreased POD activity, especially in Zizania latifolia. The proline content increased more evidently in Sagittaria sagittifolia than that in Zizania latifolia, and the CAT activity showed no significant change in both plants. Removals of TN, NH4+-N and TP increased with increasing degree of eutrophication, reaching the highest value 96.5%, 97.2% and 92.7%, respectively, in Sagittaria sagittifolia under weighty eutrophic water. However, Zizania latifolia achieved its highest remove efficiency of TN and NH4+-N under moderate eutrophic water and that of TP under weighty eutrophic water. Besides, TN, NH4+-N and TP removals were higher in Zizania latifolia than those in Sagittaria sagittifolia under mild eutrophic water. These results suggested that Zizania latifolia and Sagittaria sagittifolia were the recommended aquatic plants in phytoremediation of mild and weighty eutrophicated water, respectively.

Parasitism by the endoparasitoid wasp Pteromalus puparum causes alterations in the plasma proteins of Pieris rapae. Analysis of plasma proteins using a proteomic approach showed that seven proteins were differentially expressed in the host pupae after 24-h parasitism. They were masquerade-like serine proteinase homolog (MSPH), enolase (Eno), bilin-binding protein (BBP), imaginal disc growth factor (IDGF), ornithine decarboxylase (ODC), cellular retinoic acid binding protein (CRABP), and one unknown function protein. The full length cDNA sequences of MSPH, Eno, and BBP were successfully cloned using rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR). Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis indicated that the transcript levels of MSPH and BBP in the fat bodies of host pupae were inducible in response to the parasitism and their variations were consistent with translational changes of these genes after parasitism, while the transcript levels of Eno and IDGF were not affected by parasitism. This study will contribute to the better understanding of the molecular bases of parasitoid-induced host alterations associated with innate immune responses, detoxification, and energy metabolism.

Parasitoid venom is a focal research point in the biological control area, which aims to explore its physiological functions and nature.Pteromalus puparumis a gregarious pupal endoparasitoid wasp which has evolved unique means to adopt the host's immune system, as no other parasitoid-associated factors other than venom are injected into its hosts during oviposition. It represents an excellent model for research of parasitoid venom. In this paper, information was gathered on outcomes ofP. puparumvenom. We first began this paper by examining its functional properties. Next, we reviewed the nature of this parasitoid's venom components. Even great achievements have been made, further research is required to uncover the sophisticated bioactivity of the venom and isolate more novel toxic peptides/proteins.

Lessons from organophosphorus pesticides, which could be bioaccumulated in non-target organisms at different trophic levels and caused unexpected negative impacts, necessitate a study of the possibility of biotransfer and bioaccumulation of Bacillus thuringiensis (Bt) insecticidal toxin(s) expressed in Bt plants. Using ELISA, we evaluated the transfer of Cry1Ab toxin in a food chain of Bt rice (KMD1 and KMD2), the target insect, Cnaphalocrocis medinalis, and its predator, Pirata subpiraticus. Cry1Ab was detected in C. medinalis and P. subpiraticus. However, the concentration of Cry1Ab detected from C. medinalis and P. subpiraticus did not increase as feeding or preying time increased. A binding study of Cry1Ab to the brush border membrane vesicle of C. medinalis and P. subpiraticus indicated that P. subpiraticus does not have binding receptors in its midgut to Cry1Ab, while C. medinalis does. Survivorship and fecundity of P. subpiraticus preying on Bt rice-fed C. medinalis were not significantly different from those preying on non-Bt rice-fed C. medinalis. Developmental time of P. subpiraticus was significantly longer when it preyed on Bt rice-fed C. medinalis than on non-Bt rice-fed prey. However, a 3-year field trial indicated that Bt rice did not significantly affect the density of P. subpiraticus.

Using electron microscopy, we demonstrated the presence of a novel small RNA-containing virus in the venom apparatus of an endoparasitic wasp, Pteromalus puparum, designated as PpSRV. Morphologically, PpSRV has classical features of picorna-like viruses. It caused the development deterioration and tissue destruction of P. puparum venom apparatus. The laboratory colony of P. puparum was infectious with PpSRV at a low level. Partial genomic sequence analysis indicated that PpSRV 3'-proximal genome was characteristically similar to most dicistroviruses containing an internal ribosome entry site (IRES) for facilitating cap-independent translation and a 3' ORF encoding a structural protein with three major capsid polyproteins (28, 31 and 28 kDa) and a minor one (9.6 kDa). Phylogenetic evolution analysis of PpSRV structural protein with the counterparts of other dicistroviruses exhibited that PpSRV belonged to the Cripavirus genus and fell into same clade with Black queen cell virus (BQCV), an infectious virus to honey bee, suggesting that PpSRV might be pathogenic to P. puparum.

The last meal of sarcophagous maggots may be useful in identifying the species on whose flesh they have fed (the"host" species). The DNA profile of the host species may indeed be detectable in the"last meal" . In this paper, mitochondrial DNA analysis of gut contents was used to identify the prior host of post-feeding larvae of Aldrichina grahami (Aldrich) (Diptera: Calliphoridae). A modified logistic equation was fitted to estimate the probability of identifying the host under five different constant temperatures (16, 20, 24, 28 and 32 o C). Our results shows that the detected time ranged from a maximum of 24 h at 32 o C to 42 h at 16 o C and a minimum of 12 h at 32 o C to 30 h at 16 o C. Furthermore, the host detection time was also calculated to give the maximal time after larval hatching from the egg. These results indicate that, in criminal cases where the maggots stray from the corpse, the last meal of the larvae should not be overlooked as potentially critical evidence.

Background: Insect hosts have evolved immunity against invasion by parasitoids, and in co-evolutionary response parasitoids have also developed strategies to overcome host immune systems. The mechanisms through which parasitoid venoms disrupt the promotion of host immunity are still unclear. We report here a new mechanism evolved by parasitoid Pteromalus puparum, whose venom inhibited the promotion of immunity in its host Pieris rapae (cabbage white butterfly). Methodology/Principal Findings: A full-length cDNA encoding a C-type lectin (Pr-CTL) was isolated from P. rapae. Quantitative PCR and immunoblotting showed that injection of bacterial and inert beads induced expression of Pr-CTL, with peaks of mRNA and Pr-CTL protein levels at 4 and 8 h post beads challenge, respectively. In contrast, parasitoid venom suppressed Pr-CTL expression when co-injected with beads, in a time and dose-dependent manner. Immunolocalization and immunoblotting results showed that Pr-CTL was first detectable in vesicles present in cytoplasm of granulocytes in host hemolymph, and was then secreted from cells into circulatory fluid. Finally, the secreted Pr-CTL bound to cellular membranes of both granulocytes and plasmatocytes. Injection of double-stranded RNA specific for target gene decreased expression of Pr-CTL, and a few other host immune-related genes. Suppression of Pr-CTL expression also down-regulated antimicrobial and phenoloxidase activities, and reducing phagocytotic and encapsulation rates in host. The inhibitory effect of parasitoid venom on host encapsulation is consistent with its effect in suppressing Pr-CTL expression. Binding assay results showed that recombinant Pr-CTL directly attached to the surface of P. puparum egges. We infer that Pr-CTL may serve as an immune signalling co-effector, first binding to parasitoid eggs, regulating expression of a set of immune-related genes and promoting host immunity. Conclusions/Significance: P. puparum venom inhibits promotion of host immune responses by silencing expression of host C-type lectin gene Pr-CTL, whose expression affected transcription of other host immune-related genes.

Selected aspects of the biology of Glyptapanteles flavicoxis (Marsh) were investigated. A nutrient source like honey was found to be an important influence on adult parasitoid longevity. A series of experiments were conducted to determine the influence of adult parental sex ratio and female density on progeny production and sex ratio. Neither had a significant influence. Zusammenfassung Zum Einflus von Zuchtbedingungen auf die Lebensdauer und die Vermehrung von Glyptapanteles flavicoxis (Marsh) Es wurden ausgewahlte Aspekte zur Biologie von G. flavicoxis untersucht und herausgefunden, das Nahrungsquellen wie z. B. Honig, einen wichtigen Einflus auf die Lebensdauer adulter Parasitoide haben. In einer Versuchsreihe sollte der Einflus des Geschlechterverhaltnisses der Elterngeneration sowie der Weibchenmenge auf die Anzahl der Nachkommenschaft und deren Geschlechterverhaltnis untersucht werden; jedoch konnte in keinem Fall ein signifikanter Einflus nachgewiesen werden.

SUMMARY The mean total progeny produced by female Glyptapanteles flavicoxis was significantly greater when the parasitoid attacked 4th instar gypsy moth rather than younger larvae. The development of parasitoids within older instars was faster than that in younger hosts. The proportion of female progeny was not altered by the host instar parasitised. The total reproductive output per female G. flavicoxis increased as the number of host larvae increased. However, since the reproductive output was spread among more available hosts, the reproductive output per host declined as the number of host larvae increased. There was no trend in the relationship between total progeny produced or the proportion of female progeny and the number of matings of parental females.