Control Of Agricultural Insect Pests Research Articles

Knockdown of MAPK p38-linked genes increases the susceptibility of Chilo suppressalis larvae to various transgenic Bt rice lines

Bacillus thuringiensis (Bt) toxins have provided exceptional control of agricultural insect pests, however, over reliance on the proteins would potentially contribute to the development of field tolerance. Developing new sustainable insect pest control methods that target the mechanisms underlying Bt tolerance can potentially support the Bt control paradigm while also providing insights into basic insect physiology. The MAPK p38 pathway is strongly associated with Bt tolerance in Chilo suppressalis, a major pest of rice. To gain insights into how this pathway impacts tolerance, high-throughput screening of C. suppressalis larval midguts initially identified eight novel target genes. Increased larval sensitivity to the transgenic cry1Ca rice strain T1C-19 was observed following RNA interference-mediated knockdown of four of the genes, Cscnc, Csgcp, Cszfp26 and CsZMYM1. Similar enhanced sensitivity to the TT51 (expressing Cry1Ab/1Ac) and T2A-1 (expressing Cry2Aa) transgenic rice lines occurred when Cszfp26 and CsZMYM1 were knocked down. All four target genes are downstream of the MAPK p38 pathway but do not participate in negative feedback loop of the pathway. These results implicate Cscnc, Csgcp, Cszfp and CsZMYM1 in the C. suppressalis transgenic cry1Ca rice tolerance mechanism regulated by MAPK p38. These findings further enhance our understanding of the MAPK p38-dependent molecular mechanisms underlying Bt tolerance in C. suppressalis and open new avenues of tolerance management to develop.

Efficiency of temperature and storage duration on some morphological measurements and reproductive capacity of the entomopathogenic nematode Heterorhabditis bacteriophora Poinar, 1976 (Rhabditida: Heterorhabditidae)’s Turkish HBH hybrid strain

Entomopathogenic nematodes (EPNs) are successfully used in the biological control of agricultural insect pests. This study aims to determine the body length of hermaphrodite individuals, egg diameter and reproductive capacity obtained from Infective Juveniles (IJs) stored at different temperatures and durations. Heterorhabditis bacteriophora Poinar, 1976 (Rhabditida: Heterorhabditidae)’s Hybrid Strain HBH was used in the study. IJs stored at 15, 25 and 35°C for 7, 14 and 21 days were inoculated onto Galleria mellonella L., 1758 (Lepidoptera: Pyralidae) last instar larvae at a dose of 100 IJs. On the 2nd day of infection, hermaphrodite individuals and eggs were obtained by dissecting the larvae. The reproductive capacity was determined 10-12 days after infection. The study was conducted in Bursa Uludağ University, Faculty of Agriculture, Plant Protection Department, Nematology Laboratory in 2023. In conclusion, the longest hermaphrodite individuals and egg diameter were obtained as 6207.22 µm and 55.65 µm, respectively from the IJs stored for 7 days at 15°C. The highest reproductive capacity was also observed as 167.500 IJs per G. mellonella larva in IJs stored under the same conditions with respect to temperature and time. This study is important for assessing the morphological effects of different temperature values and storage durations on EPNs.

The gut symbiont Sphingomonas mediates imidacloprid resistance in the important agricultural insect pest Aphis gossypii Glover

BackgroundNeonicotinoid insecticides are applied worldwide for the control of agricultural insect pests. The evolution of neonicotinoid resistance has led to the failure of pest control in the field. The enhanced detoxifying enzyme activity and target mutations play important roles in the resistance of insects to neonicotinoid resistance. Emerging evidence indicates a central role of the gut symbiont in insect pest resistance to pesticides. Existing reports suggest that symbiotic microorganisms could mediate pesticide resistance by degrading pesticides in insect pests.ResultsThe 16S rDNA sequencing results showed that the richness and diversity of the gut community between the imidacloprid-resistant (IMI-R) and imidacloprid-susceptible (IMI-S) strains of the cotton aphid Aphis gossypii showed no significant difference, while the abundance of the gut symbiont Sphingomonas was significantly higher in the IMI-R strain. Antibiotic treatment deprived Sphingomonas of the gut, followed by an increase in susceptibility to imidacloprid in the IMI-R strain. The susceptibility of the IMI-S strain to imidacloprid was significantly decreased as expected after supplementation with Sphingomonas. In addition, the imidacloprid susceptibility in nine field populations, which were all infected with Sphingomonas, increased to different degrees after treatment with antibiotics. Then, we demonstrated that Sphingomonas isolated from the gut of the IMI-R strain could subsist only with imidacloprid as a carbon source. The metabolic efficiency of imidacloprid by Sphingomonas reached 56% by HPLC detection. This further proved that Sphingomonas could mediate A. gossypii resistance to imidacloprid by hydroxylation and nitroreduction.ConclusionsOur findings suggest that the gut symbiont Sphingomonas, with detoxification properties, could offer an opportunity for insect pests to metabolize imidacloprid. These findings enriched our knowledge of mechanisms of insecticide resistance and provided new symbiont-based strategies for control of insecticide-resistant insect pests with high Sphingomonas abundance.

Exploration of the DNA Photocleavage Activity of O-Halo-phenyl Carbamoyl Amidoximes: Studies of the UVA-Induced Effects on a Major Crop Pest, the Whitefly Bemisia tabaci

The DNA photocleavage effect of halogenated O-carbamoyl derivatives of 4-MeO-benzamidoxime under UVB and UVA irradiation was studied in order to identify the nature, position, and number of halogens on the carbamoyl moiety that ensure photoactivity. F, Cl, and Br-phenyl carbamate esters (PCME) exhibited activity with the p-Cl-phenyl derivative to show excellent photocleavage against pBR322 plasmid DNA. m-Cl-PCME has diminished activity, whereas the presence of two halogen atoms reduced DNA photocleavage. The substitution on the benzamidoxime scaffold was irrelevant to the activity. The mechanism of action indicated function in the absence of oxygen, probably via radicals derived from the N-O bond homolysis of the carbamates and in air via hydroxyl radicals and partially singlet oxygen. The UVA-vis area of absorption of the nitro-benzamidoxime p-Cl-PCMEs allowed for the investigation of their potential efficacy as photopesticides under UVA irradiation against the whitefly Bemisia tabaci, a major pest of numerous crops. The m-nitro derivative exhibited a moderate specificity against the adult population. Nymphs were not affected. The compound was inactive in the dark. This result may allow for the development of lead compounds for the control of agricultural insect pests that can cause significant economic damage in crop production.

Biological Activity of Cupressus sempervirens Essential Oil.

The aim of this study was to evaluate the antioxidant, antibiofilm, antimicrobial (in situ and in vitro), insecticidal, and antiproliferative activity of Cupressus sempervirens essential oil (CSEO) obtained from the plant leaf. The identification of the constituents contained in CSEO was also intended by using GC and GC/MS analysis. The chemical composition revealed that this sample was dominated by monoterpene hydrocarbons α-pinene, and δ-3-carene. Free radical scavenging ability, performed by using DPPH and ABTS assays, was evaluated as strong. Higher antibacterial efficacy was demonstrated for the agar diffusion method compared to the disk diffusion method. The antifungal activity of CSEO was moderate. When the minimum inhibitory concentrations of filamentous microscopic fungi were determined, we observed the efficacy depending on the concentration used, except for B. cinerea where the efficacy of lower concentration was more pronounced. The vapor phase effect was more pronounced at lower concentrations in most cases. Antibiofilm effect against Salmonella enterica was demonstrated. The relatively strong insecticidal activity was demonstrated with an LC50 value of 21.07% and an LC90 value of 78.21%, making CSEO potentially adequate in the control of agricultural insect pests. Results of cell viability testing showed no effects on the normal MRC-5 cell line, and antiproliferative effects towards MDA-MB-231, HCT-116, JEG-3, and K562 cells, whereas K562 cells were the most sensitive. Based on our results, CSEO could be a suitable alternative against different types of microorganisms as well as suitable for the control of biofilms. Due to its insecticidal properties, it could be used in the control of agricultural insect pests.

RNAi-mediated knockdown of arginine kinase genes leads to high mortality and negatively affect reproduction and blood-feeding behavior of Culex pipiens pallens.

Arginine kinase (AK) is one of the crucial enzymes involved in energy metabolism in invertebrates, and has been proposed as the target for RNA interference (RNAi)-based control of agricultural insect pests. While there is only one AK gene in most insects, two AK genes were identified in Culex pipiens pallens, the primary vector of lymphatic filariasis and epidemic encephalitis. The full-length cDNA sequences of CpAK1 and CpAK2 genes were obtained by reverse transcription PCR(RT-PCR) and rapid amplification of cDNA ends (RACE). The expression levels of CpAK1 and CpAK2 in different developmental stages and tissues were detected by reverse transcription quantitative PCR (RT-qPCR). The role of CpAK1 and CpAK2 in the reproduction and blood feeding behavior was analyzed using RNA interference (RNAi). Full-length cDNAs of CpAK1 and CpAK2 were isolated from Cx. pipiens pallens. Analysis of the expression pattern revealed that the mRNA level of CpAK1 was significantly higher than CpAK2 in all development stages and tissues examined, and the expressions of both CpAK1 and CpAK2 were upregulated in response to blood feeding. The co-knockdown of CpAK1 and CpAK2 mediated by RNAi led to high mortality (74.3%) of adult female mosquitoes and decreased hatchability (59.9%). Remarkably, the blood feeding rate and the engorgement rate of the female mosquitoes were negatively affected by co-injection of dsRNAs targeting CpAK1 and CpAK2. CpAK1 and CpAK2 were detected in all developmental stages and tissues, but showed divergence in expression level. RNAi-mediated knockdown of AK genes leads to high mortality and negatively affect blood-feeding behavior of Cx. pipiens pallens, suggesting that AK could be used for the target of RNAi-based mosquito control in the future.

Diallyl trisulfide reduced the reproductive capacity of male Sitotroga cerealella via the regulation of juvenile and ecdysone hormones

Environmental pollution and resistance in animals are major concerns for the application of synthetic pesticides. Diallyl trisulfide (DAT), an active compound in garlic essential oil, is a novel tool for active and safe control of agricultural insect pests. In this study, we analysed the effects of DAT (0.01 μL/L) on the protein content in male reproductive tissues (accessory glands, ejaculatory ducts, and testis), and juvenile hormone (JH) and ecdysone titres in a highly detrimental pest of stored products, Sitotroga cerealella. Evaluation of the expression profile of JH and ecdysone pathway-related genes in various tissues indicated that the accessory gland protein and ecdysone titres were markedly decreased after DAT fumigation, whereas the testis protein content and JH titre were increased. However, the protein content of the ejaculatory ducts remained unchanged between the treated and control groups. Further investigation revealed that DAT disrupted the mRNA expression of key enzymes involved in JH and ecdysone pathways. While increased mRNA levels of juvenile hormone acid O-methyltransferase (JHMAT) and Kruppel homologue 1 (Kr-h1) were observed after 4 and 7 h of DAT fumigation, the levels of juvenile hormone epoxide hydrolase (JHEH) were substantially reduced 3 h post-fumigation. mRNA levels of the ecdysone-responsive gene, FTZF1, and cytochrome P450 enzyme, CYP315A1, were notably decreased at 7 h and 4 h, respectively, post-fumigation, whereas CYP314A1 and CYP302A1 mRNA levels decreased after 3 h and 4 h, respectively. While DAT fumigation disrupted sperm number in the testis, ejaculatory ducts, and seminal vesicles, topical application of the 20-hydroxyecdysone (20E) analogue also lowered sperm number in the ejaculatory ducts. Topical application of methoprene, a JH analogue, increased the protein content in the testes, but not in the accessory glands or ejaculatory ducts. However, the survival rate was not affected by the topical application of methoprene or 20E. These data suggest that DAT regulates JH and ecdysone via its molecular pathway genes and modulates endocrine secretion during the male reproductive process.

Identification and Functional Characterization of a Novel Insecticidal Decapeptide from the Myrmicine Ant Manica rubida.

Ant venoms contain many small, linear peptides, an untapped source of bioactive peptide toxins. The control of agricultural insect pests currently depends primarily on chemical insecticides, but their intensive use damages the environment and human health, and encourages the emergence of resistant pest populations. This has promoted interest in animal venoms as a source of alternative, environmentally-friendly bio-insecticides. We tested the crude venom of the predatory ant, Manica rubida, and observed severe fitness costs in the parthenogenetic pea aphid (Acyrthosiphon pisum), a common agricultural pest. Therefore, we explored the M. rubida venom peptidome and identified a novel decapeptide U-MYRTX-MANr1 (NH2-IDPKVLESLV-CONH2) using a combination of Edman degradation and de novo peptide sequencing. Although this myrmicitoxin was inactive against bacteria and fungi, it reduced aphid survival and reproduction. Furthermore, both crude venom and U-MYRTX-MANr1 reversibly paralyzed injected aphids and induced a loss of body fluids. Components of M. rubida venom may act on various biological targets including ion channels and hemolymph coagulation proteins, as previously shown for other ant venom toxins. The remarkable insecticidal activity of M. rubida venom suggests it may be a promising source of additional bio-insecticide leads.

Economic Entomology in the 21st Century, as Seen from a Bibliometric Analysis of Two of Its Leading Journals, 2000–2018. Part IV: Beneficial Arthropods Used In the Biological Control of Agricultural Insect Pests

ABSTRACTAn analysis of two leading journals for economic entomologists employing beneficial arthropods against agricultural insect pests --- the Journal of Economic Entomology, and Biological Control--- uncovered a highly concordant distribution of species used and species attacked. Parasitizing wasps were by far the most common beneficials. These were followed in frequency by predatory versions of the following: true bugs, mites, lady beetles, and lacewings. Assemblages of larger ground beetles and spiders provided good crop protection in fewer but still consequential dilemmas. Familiarity with prominent species will help science librarians who are not entomologists by training, but who may serve students and faculty in agriculture, ecology, or environmental studies, by raising their own awareness of this important means of pest control with less reliance on potentially toxic insecticides.

Public attitudes toward the presence and management of bats roosting in buildings in Great Smoky Mountains National Park, Southeastern United States

In the human dimensions of wildlife management, evaluating stakeholder perceptions of target species helps inform effective conservation efforts. Stakeholder perceptions are invaluable when managing taxa like bats, which may have historically negative cultural preconceptions. However, insectivorous bats provide critical ecosystem services in North America through agricultural insect pest control, and many of these species are threatened by white-nose syndrome (WNS), a disease caused by an invasive fungal pathogen. In Great Smoky Mountains National Park (GRSM), the most visited National Park in the United States (USA), bats are regularly observed roosting in historical buildings by visitors and park employees during summer. As a result, natural and cultural resource managers seek to ensure public safety and protect historical structures while minimizing impacts on bats, especially in light of declines in bat populations as a result of WNS. However, managers lacked information on visitor perceptions of bats and support for potential management action regarding the taxon. From June to August 2016, we surveyed 420 park visitors at three sites in the Cades Cove area of GRSM on their attitudes toward bats, perception of threats to and ecosystem services provided by bats, and support for management of bats. Most respondents supported management action to protect bats in buildings in Cades Cove during summer (76%). Standardized parameter estimates from a multiple linear regression developed with survey data indicated that attitudes toward bats and perception of threats to bats had the greatest effects on support for bat management. Wildlife management and conservation agencies seeking to further cultivate support for management of bats roosting in public spaces may apply these results in the design of tailored programming and outreach materials.

Agricultural production: assessment of the potential use of Cas9-mediated gene drive systems for agricultural pest control

ABSTRACTTo highlight how gene drives could be useful for control of agricultural insect pests, we selected species that are pests of animals (New World screwworm), plants (spotted wing Drosophila, diamondback moth, Bemisia tabaci whitefly), or stored grains (red flour beetle). With the exception of whitefly, routine methods for delivering DNA to the germline and selecting for genetically modified insects have been developed. The traditional approach in agriculture has been to suppress insect pest populations using insecticides and other farming practices. Similarly, we suggest the main use of gene drives in agriculture will be for population suppression through targeting essential genes. We provide examples of gene drives that target specific genes including female-essential genes. Further, we discuss issues related to containment in the laboratory and eventual field testing of strains harboring a Cas9-mediated gene drive system.

Evaluation of the growth and sporulation of different entomopathogenic fungi in different liquid and solid media at varied concentrations

Entomopathogenic fungi like Beauveria bassiana, Metarhizium anisopliae and Lecanicillium lecanii are used in biological control of agricultural insect pests. Their specific mode of action makes them an effective alternative to the chemical Insecticides. Virulent strains of Entomopathogenic fungi are effectively formulated and used as bio-insecticides world-wide. Amenable and economical multiplication of a virulent strain in a large scale is important for them to be useful in the field. Culture media plays a major role in the large-scale multiplication of virulent strains of Entomopathogens. Different substrates and media components are being used for this purpose. Yet, each strain differs in its nutritional requirements for the maximum growth and hence it is necessary to standardize the right components and their optimum concentrations in the culture media for a given strain of Entomopathogen. In the current study, three different nitrogen sources and two different carbon sources were tried to standardize the mass multiplication media for seven test isolates of Entomopathogenic fungi. A study was also conducted to determine the ideal grain media for the optimum conidial yields of the test isolates. Yeast extract was found to be the best Nitrogen source for the isolates. The isolates tested, differed in their nutritional requirements and showed variation in the best nitrogen and carbon sources necessary for their growth. Variation was also found in the optimum concentration of both the ingredients for the growth and sporulation of the isolates. In the solid-state fermentation study, rice was found to be the best grain for the growth of most of the fungi followed by barley. The significance of such a study in the development of an effective Myco-insecticide is vital and can be successfully employed in agriculture is discussed.

RNA interference as a gene silencing tool to control Tuta absoluta in tomato (Solanum lycopersicum).

RNA interference (RNAi), a gene-silencing mechanism that involves providing double-stranded RNA molecules that match a specific target gene sequence, is now widely used in functional genetic studies. The potential application of RNAi-mediated control of agricultural insect pests has rapidly become evident. The production of transgenic plants expressing dsRNA molecules that target essential insect genes could provide a means of specific gene silencing in larvae that feed on these plants, resulting in larval phenotypes that range from loss of appetite to death. In this report, we show that the tomato leafminer (Tuta absoluta), a major threat to commercial tomato production, can be targeted by RNAi. We selected two target genes (Vacuolar ATPase-A and Arginine kinase) based on the RNAi response reported for these genes in other pest species. In view of the lack of an artificial diet for T. absoluta, we used two approaches to deliver dsRNA into tomato leaflets. The first approach was based on the uptake of dsRNA by leaflets and the second was based on “in planta-induced transient gene silencing” (PITGS), a well-established method for silencing plant genes, used here for the first time to deliver in planta-transcribed dsRNA to target insect genes. Tuta absoluta larvae that fed on leaves containing dsRNA of the target genes showed an ∼60% reduction in target gene transcript accumulation, an increase in larval mortality and less leaf damage. We then generated transgenic ‘Micro-Tom’ tomato plants that expressed hairpin sequences for both genes and observed a reduction in foliar damage by T. absoluta in these plants. Our results demonstrate the feasibility of RNAi as an alternative method for controlling this critical tomato pest.

Insight into the feeding behavior of predatory mites on Beauveria bassiana, an arthropod pathogen.

Interactions between fungal entomopathogens and pest predators are particularly relevant in control of agricultural insect pests. In a laboratory study, we confirmed that the predatory mite, Neoseiulus barkeri, exhibited feeding behavior on the entomopathogenic fungus Beauveria bassiana conidia through DNA extracts. Using transmission electron microscopy, we determined that the majority of conidia found in the mite gut tended to dissolve within 24 h post ingestion, suggesting that the conidia had probably lost their viability. To our knowledge this is the first report of feeding behavior of phytoseiid mites on entomopathogenic fungus. The findings expand our knowledge of fungus–predator interactions.

Endosymbiotic bacteria in insects: guardians of the immune system?

Insects have evolved obligate, mutualistic interactions with bacteria without further transmission to other eukaryotic organisms. Such long-term obligate partnerships between insects and bacteria have a profound effect on various physiological functions of the host. Here we provide an overview of the effects of endosymbiotic bacteria on the insect immune system as well as on the immune response of insects to pathogenic infections. Potential mechanisms through which endosymbionts can affect the ability of their host to resist an infection are discussed in the light of recent findings. We finally point out unresolved questions for future research and speculate how the current knowledge can be employed to design and implement measures for the effective control of agricultural insect pests and vectors of diseases.

Flexible metabolism in Metarhizium anisopliae and Beauveria bassiana: role of the glyoxylate cycle during insect pathogenesis

Insect pathogenic fungi such as Metarhizium anisopliae and Beauveria bassiana have an increasing role in the control of agricultural insect pests and vectors of human diseases. Many of the virulence factors are well studied but less is known of the metabolism of these fungi during the course of insect infection or saprobic growth. Here, we assessed enzyme activity and gene expression in the central carbon metabolic pathway, including isocitrate dehydrogenase, aconitase, citrate synthase, malate synthase (MLS) and isocitrate lyase (ICL), with particular attention to the glyoxylate cycle when M. anisopliae and B. bassiana were grown under various conditions. We observed that ICL and MLS, glyoxylate cycle intermediates, were upregulated during growth on 2-carbon compounds (acetate and ethanol) as well as in insect haemolymph. We fused the promoter of the M. anisopliae ICL gene (Ma-icl) to a marker gene (mCherry) and showed that Ma-icl was upregulated when M. anisopliae was grown in the presence of acetate. Furthermore, Ma-icl was upregulated when fungi were engulfed by insect haemocytes as well as during appressorium formation. Addition of the ICL inhibitor 3-nitroproprionate delayed conidial germination and inhibited appressorium formation. These results show that these insect pathogenic fungi have a flexible metabolism that includes the glyoxylate cycle as an integral part of germination, pathogenesis and saprobic growth.

From microbial sprays to insect-resistant transgenic plants: history of the biospesticideBacillus thuringiensis. A review

Bacillus thuringiensis, known as Bt, is a spore-forming bacterium that occurs naturally in soil and that produces highly specific insecticidal proteins called Cry proteins. These proteins are stomach poisons that specifically affect insects. Today, Bt preparations are considered as the most effective, specific and environmentally-friendly bioinsecticides; they have been used as biological pesticides in agriculture, forestry and in human health for the elimination of vectors of diseases for more than 60 years and their implementation far exceeds other microbial agents such as fungi, protozoa or viruses. This review on the use of this entomopathogenic bacterium in crop protection is not intended to be a compilation of the results of all the investigations made in this field. Instead, it is an attempt to provide an overview of the major trends and developments of Bt for the control of agricultural insect pests and to describe the main approaches that have been used to improve this natural bioinsecticide. Bt-based insecticides are considered safe for mammals and birds, and are safer for non-target insects than conventional insecticides; they have become the most widely used microbial insecticides. However, Bt products have several limitations, such as a narrow activity spectrum, instability in rain and sunlight, and inefficiency against pest feeding on internal tissues of the plants. The first step towards improving Bt has involved the isolation of new strains with higher and broader insecticidal activity against targeted insect pests and the cloning of cry genes encoding new insecticidal crystal proteins. A second strategy was to increase the persistence of its toxins in the field by encapsulation in recombinant asporogenic Bt strains or other heterologous recombinant microbial hosts; this protected the toxins against UV degradation and had the advantage that the transgenic microorganisms released into the environment were non-viable. Bt has also become a key source of genes for transgenic expression to provide pest resistance in plants and in so-called genetically modified plants. The engineering of plants to express Bt cry genes has been especially helpful against pests that attack parts of the plant that are usually not well protected by conventional insecticide application. The potential effects on human health and the environment of the large-scale use of these Bt crops are also in the scope of this review.

A review of encyrtid wasps (Hymenoptera, Chalcidoidea, Encyrtidae) of Macaronesia

The review contains an annotated list of 62 species of Encyrtidae from Macaronesia with data on their distribution, hosts, and use in biological control of agricultural insect pests. An attempt is undertaken to classify most of them according to nine types of ranges and to separate species which were introduced to Macaronesia by the ecesis, i.e., accidentally with their hosts. Three genera are synonymized: Acerophagus E.A. Smith, 1880 (= Pseudaphycus Timberlake, 1916, syn. n.); Isodromus Howard, 1887 (= Eupoecilopoda Erdos et S. Novicky, 1955, syn. n.); Syrphophagus Ashmead, 1900 (= Tassonia Girault, 1921, syn. n.). Cerchysiella togashii Tachikawa, 1988 is placed in the genus Zaommoencyrtus Girault, 1916. Cerchysiella koenigsmanni (Trjapitzin, 1985) is considered a subjective synonym of C. latiscapus (Shafee et Fatma, 1984). The following replacement names are given: Acerophagus xuzhihongi Trjapitzin, nom. n. for A. clavatus Xu Zhihong, 1999 non A. clavatus Pilipjuk, 1981; and Anagyrus sergueii Trjapitzin, nom. n. for Anagyrus orbitalis Timberlake, 1941 non A. orbitalis (Ruschka, 1923). Material on encyrtids from the Macaronesian islands Tenerife and Madeira in the museums in Madrid, Helsinki and Washington were examined.

Conjugation of δ‐endotoxin from Bacillus thuringiensis with abamectin of Streptomyces avermitilis as a new type of biocide, GCSC‐BtA, for control of agricultural insect pests

AbstractConjugation of δ‐endotoxin from Bacillus thuringiensis with abamectin, a toxin of Streptomyces avermitilis, was carried out to form a new type of biocide, GCSC‐BtA based on “Germany‐China Scientific Cooperation” research, for the control of agricultural insect pests. The strategy for biochemical linkage was designed by conjugating an amino group in B.t. protoxin with a carboxyl group in carboxylated abamectin under the treatment of conjugator EDC [1‐Ethyl‐3‐(3‐dimethylaminopropyl carbodiimide Hydrochloride)]. The formation of B.t. protoxin was processed by solubilizing B.t. crystal in 25 mM dithiothreitol (DTT) at 37 °C for 2 h. The carboxylated abamectin was formed by carboxylating the NaH‐activated abamectin with 10 mg/ml butyric anhydride at 111 °C in a water‐circumfluent condensation device for 2 h. The conjugating reaction, consisting of 5 mg/ml B.t. protoxin, 10 mg/ml carboxylated abamectin and 19.17 mg/ml EDC, was successfully conducted at room temperature for 24 h. Significant differences were found between pure abamectin, carboxylated abamectin and the conjugated BtA by means of UV‐photo absorptions recorded at wavelengths 354, 438, 518, 600 nm (P < 0.01). LT50 of the conjugated GCSC‐BtA to the 3rd instar larvae of Plutella xylostella (L.) (Lep., Plutellidae) was 35.27 μg a.i./ml, about 62 % and 76 % of that caused by the B.t. protoxin and the caxboxylated abamectin, respectively. The conjugated GCSC‐BtA caused 87.14 % mortalities in larvae of P. xylostella, 93.75 % in adult Myzus persicae (Sulzer) (Hom., Aphididae) and 89.33 % in adult Phyllotreta vittata Fabricius (Col., Chrysomelidae) as compared to 48.33 % by the B.t. crystal only in P. xylostella. The symptoms caused by conjugated GCSC‐BtA in the 3rd instar of P. xylostella were black color in the head part and white‐yellow in the abdomen of dead larvae, which differed from the black color or the white‐yellow all along the body caused by either the B.t. crystal or the abamectin, respectively. It was concluded that the conjugated GCSC‐BtA biocide had a broader host spectrum and a faster killing speed than either the B.t. crystal or abamectin alone for the control of agricultural pests.

Análise da riqueza da fauna de Braconidae (Hymenoptera, Ichneumonoidea) em remanescentes naturais da Área de Proteção Ambiental (APA) de Descalvado, SP

Analysis of the Braconidae (Hymenoptera: Ichneumonoidea) fauna richness in natural remnants of the Area de Protecao Ambiental (APA) of Descalvado, SP. A survey of the Braconidae fauna on fragmentary natural environments at northeast Sao Paulo State was conducted using Malaise traps installed in five sites of Area de Protecao Ambiental of Descalvado. A total of 2,262 specimens, representing 22 subfamilies and 94 genera, was sampled throughout a period of sixteen months (from May, 1999 to August, 2000). Biological data were obtained from current literature and behavior patterns of host utilization for the studied fauna, mainly Braconidae which have been successful in control of agricultural insect pests. Statistical analyses indicated that the asymptote of the genera richness has been approached using this sampling method. This is the most complete survey yet available from natural fragmentary areas of the Descalvado Braconidae fauna. Patterns of distribution and richness of the Braconidae genera were established. Cluster Analysis was adopted taking as attribute the number of Braconidae genera.