Cryptic species of insect disease vectors and their importance.
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Animals↗
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Immunoaffinity purification of the neuropeptide prothoracicotropic hormone from Manduca sexta.
The prothoracicotropic hormones (PTTH) are cerebral peptides that control insect postembryonic development by stimulating the prothoracic glands to synthesize ecdysteroids. Using immunoaffinity chromatography and SDS-PAGE, a 25.5 kDa big PTTH has been purified from Manduca sexta. Based upon SDS-PAGE and Western blot analysis, the native form of big PTTH appears to be a dimer with monomers of 16.5 kDa. Four HPLC-separated fragments of this acidic peptide were sequenced and exhibited no sequence similarity with Bombyx mori PTTH. In agreement with this finding, the basic Bombyx PTTH had no PTTH bioactivity in Manduca. One sequenced fragment of the Manduca PTTH is approximately 70% similar to the vertebrate cellular retinoid binding proteins, suggesting these binding proteins may be present in insects.
Amino Acid Sequence↗
The role of the chemical industry in development.
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Agriculture↗
Aircraft disinsection: an international concern for the protection of man and agriculture.
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Aircraft↗
Maserlike nonlinear scatter from human breath, a surface-enhanced far infrared scatter effect.
A cotton tip applicator was utilized to stimulate far IR scatter (emission) from human breath. The reticulated cotton tip was vibrated at 200 Hz, the acoustic frequency that matches the average vibration frequency of common species of mosquitoes. Mosquitoes have open resonator dielectric sensillae (spines) on their antenna that match the 643 cm-1 (15.5) Cabannes line in the 667 cm-1 region of CO2 rotation line, acoustic Stokes Brillouin and Stokes Raman lines are noted. Implications of such maserlike (coherent or partial coherent) scatter lines to insect control and disease diagnostics are discussed.
Animals↗
Pyrethroids, nerve poisons: how their risks to human health should be assessed.
The extensive worldwide efforts of structural modification of natural pyrethrins for better performances have resulted in successful development of a wide variety of synthetic pyrethroids with tremendously high efficacy, knock-down activity or vapor action, and/or with acceptable environmental stability and safety. Currently these pyrethroids including their preferentially manufactured stereoisomers are widely used in agriculture, and for public health as well as household insect control. The detailed toxicology and metabolism studies intended to attain human risk assessment have revealed that with voltage-dependent sodium channel as target site pyrethroids induce pronounced repetitive activity characterized grossly by tremor, hypersensitivity, choleoathetosis, and salivation. In addition, so-called cyano-pyrethroids cause transient skin paresthesia in workers. With regard to tumorigenicity, mutagenicity, teratogenicity and developmental toxicity, no significant findings have been reported. Pyrethroids are eliminated from the animals quite rapidly and completely, undergoing oxidation and ester hydrolysis followed by various conjugations, with low tissue residues. Thus, overall, sound scientific bases exist for human risk assessment under the present usage conditions.
Animals↗
A survey of medicinal plants of minas gerais. Brazil.
A list of fifty-two plant species used for medicinal purposes or to control arthropods by peasants near Belo Horizonte, Minas Gerais (Brazil) has been compiled. Preparation procedures are also noted. The use of plants for controlling insects and to heal a condition known as "ferida brava" is discussed. The role of indigenous plant remedies within the local health care system is worthy of a closer investigation.
Botany↗
Linkage of kdr-type resistance and the para-homologous sodium channel gene in German cockroaches (Blattella germanica).
Pyrethroids are an important class of insecticides for controlling insect pests, including the German cockroach. Unfortunately, many insects have developed resistance to pyrethroids. One of the most important mechanisms of resistance is kdr (knockdown resistance) which is characterized by neural insensitivity to pyrethroids and DDT. To investigate whether the voltage-dependent sodium channel is involved in kdr-type resistance in the German cockroach, we isolated a 120 bp DNA fragment of the para-homologous sodium channel gene from German cockroaches. Using this fragment as a probe, we identified a restriction fragment length polymorphism (RFLP) of the para-homologous sodium channel gene between susceptible and kdr-type resistant German cockroaches. RFLP analysis of F2 and backcross cockroach populations (total of 331 individuals) showed that all homozygous resistant individuals had a 3.7 kb EcoRI fragment, all homozygous susceptible individuals had a 3.0 kb EcoRI fragment, and all heterozygous individuals had both 3.7 and 3.0 kb fragments. No recombination was detected between the kdr-type resistance locus and the para-homologous sodium channel gene. This suggests that the kdr-type resistance locus and para-homologous sodium channel gene are identical or tightly linked (< 0.2 cM) in German cockroaches. Our results provide strong evidence that modification of para-homologous sodium channels is associated with kdr-type resistance.
Amino Acid Sequence↗
Stereospecific, mechanism-based, suicide inhibition of a cytochrome P450 involved in ecdysteroid biosynthesis in the prothoracic glands of Manduca sexta.
The first required step in ecdysteroid (molting hormone) biosynthesis, dietary cholesterol (C) conversion to 7-dehydrocholesterol (7dC) via 7,8-dehydrogenation, is mediated by a microsomal cytochrome-P450 monooxygenase specific to the larval prothoracic gland. A subsequent series of unknown "black-box" oxidations of 7dC result in the unusual ring geometry (cis-A/B) and functionality (6-keto-7-ene-14-alpha-ol) of the ecdysteroids and has been thought to involve the initial formation of alpha-5,6-epoxy-7-dehydrocholesterol (alpha epo7dC). Pharmacological studies indicated that conversion of C to 7dC in prothoracic gland homogenates was strongly and equally inhibited by the isomeric cholesterol substrate analogues alpha- and beta-5,6-epoxycholesterol (alpha- and beta epoC) and alpha- and beta-5,6-iminocholesterol (alpha- and beta iminoC). With respect to the conversion of C to ecdysteroids by disrupted glands, however, the two alpha-isomeric substrates were 10-fold more inhibitory than were their beta-analogues. Indeed, alpha amino C was as active as the non-specific pyrimidyl cytochrome-P450 monooxygenase inhibitor fenarimol that shows moderate toxicity in many insect species. All four cholesterol analogues competitively inhibited cholesterol 7,8-dehydrogenation, but only alpha epoC and possibly alpha iminoC were desaturated to delta 7-products. Although the KmS (and KiS) for all the substrates were similar (1.7-6.0 x 10(-5) M), the Vmax for alpha epoC dehydrogenation was eight-fold higher than that of C, making it a superior substrate for following this reaction in ecdysteroidogenic tissues rich in endogenous C. The 7,8-dehydrogenation of alpha epoC and alpha iminoC by prothoracic glands would produce the potentially reactive intermediates, alpha epo7dC and alpha imino7dC, respectively. They, in turn, could then undergo facile, acid-catalyzed ring-opening to the allylic-stabilized carbo-cation electrophiles. These very reactive, transient species, if formed in the active site of the monooxygenase, would then alkylate either the heme group or the apoprotein of the cytochrome or both, leading to the irreversible inhibition of the enzyme. The present data show that alpha epoC and probably alpha iminoC are mechanism-based suicide inhibitors of the enzyme catalyzing cholesterol 7,8-dehydrogenation and may be the prototypes of a new class of selective insect control agents.
Animals↗
Chagas disease parasite induces behavioural changes in the kissing bug Mepraia spinolai.
Parasites have been shown to manipulate the feeding behaviour of their invertebrate vectors, which results in an increased probability of transmission to definitive hosts. Most evidence for this hypothesis comes from protozoan species with salivary transmission but evidence for stercorarian parasite transmission is lacking. We present experimental evidence that infection of the kissing bug Mepraia spinolai (Hemiptera; Reduviidae) with the protozoan Trypanosoma cruzi reduced the time to detect potential hosts in comparison to control insects. Infected bugs bit about twice more often than uninfected nymphs and defecated 8 min after the last blood meal whereas uninfected bugs needed 11 min. The behaviour of male and female nymphs did not differ significantly. Since all of these traits relate to parasite transmission, we suggest that parasite-mediated changes in the foraging and defecation behaviour of M. spinolai may promote the spread of T. cruzi toward definitive hosts.
Animals↗
Expression patterns of three heat shock protein 70 genes among developmental stages of the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae).
Three genes were identified encoding heat shock protein 70's in Tribolium castaneum (Herbst) and they were tentatively named as tchsp70 I, tchsc70 II, and tchsp70 III. Comparison of deduced amino acid sequences of tchsp70 I and tchsc70 II showed 99% identity. However, the amino acid sequence of tchsp70 III was only 58.5% identical to those of tchsp70 I and tchsc70 II. Stage-specific expression patterns of the tchsp70 were investigated in young larvae, old larvae, pupae, and adults of T. castaneum exposed for 1 h to 23 degrees C (control) or 40 degrees C (heat-shock). Northern blot and real-time quantitative PCR analyses were carried out to determine mRNA levels in each life stage. Transcripts of all three genes were detected by Northern blotting, and the sizes were 2.4- 2.2-, and 2.3-kb for tchsp70 I, tchsc70 II, and tchsp70 III, respectively. A 1.1- to 2.0-fold increased expression of tchsp70 I mRNA was found in heat-shocked developmental stages compared with the control. The expression of tchsc70 II mRNA among developmental stages was similar between heat-shocked and control insects, and the expression of tchsp70 III mRNA varied among developmental stages. Results suggest that the expression of tchsp70 I gene is heat-inducible, tchsc70 II is constitutive, and tchsp70 III is developmentally regulated in T. castaneum.
Amino Acid Sequence↗
Dissipation of DDT in a heavily contaminated soil in Mato Grosso, Brazil.
After the prohibition of organochlorine-pesticide use in Brazil for controlling insect vector diseases, Mato Grosso State gathered the exceeding DDT and stored it irregularly in an open air area that belongs to the National Health Foundation, causing soil contamination. This study aimed to evaluate the contamination level and dissipation of p,p'-DDT and p,p'-DDE in this area. For that, surface soil samples were collected on 19 September 2000, 15 December 2000, 4 April 2001 and soil samples 30-40 cm; 60-70 cm and 90-100 cm deep were taken from five points in the studied area on 17 July 2001. The contaminants were determined by a small scale method which consists on extraction and clean-up steps combined into one step by transferring soil samples mixed with neutral alumina to a chromatographic column prepacked with neutral alumina and elution with hexane:dichloromethane (7:3 v:v). The eluate was concentrated and the analytes were quantified by gas chromatography with an electron-capture detector. p,p'-DDT at surface soil ranged from 3,800 to 7,300 mg kg(-1). 30-40 cm deep soil sample concentrations varied from 0.036 to 440 mg kg(-1) while 90-100 cm deep samples varied from 0.069 to 180 mg kg(-1). Volatilization is probably the main dissipation process. The p,p'-DDT is moving slowly downward in the soil profile, however, the levels of this contaminant are high enough to present risk to underground waters.
Brazil↗
Biological activity of neolignans on the post-embryonic development of Chrysomya megacephala.
Phytochemicals endowed with hormonal, antihormonal, or toxic activity are potential agents for insect control. The effect of some neolignans on one of the most prevalent flies in urban areas, which constitutes a menace to public health, was investigated.
Animals↗
AcMNPV ChiA protein disrupts the peritrophic membrane and alters midgut physiology of Bombyx mori larvae.
Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) chitinase A (ChiA) is a protein which promotes the final liquefaction of infected host larvae. The potential of this viral molecule as a new tool for insect control is explored here. The ChiA gene was isolated from the AcMNPV genome by PCR and expressed in E. coli. The recombinant protein, purified by affinity chromatography, showed both exo- and endo-chitinase activities and produced perforations on the peritrophic membrane (PM) of Bombyx mori larvae which increased in number and in size, in a dose-dependent manner. This structural alteration resulted into a significant increase of PM permeability to methylene blue and to the small neuropeptide proctolin. When the fifth instar larvae of B. mori were fed on a artificial diet supplemented with the recombinant ChiA, 100% mortality was observed at a dose of 1 microg/g of larval body weight (LW), while at sub-lethal doses of 0.56 microg/g LW, a reduced larval growth was recorded. These results indicate that AcMNPV-ChiA may offer interesting new opportunities for pest control.
Animals↗
Elevated activity of an Epsilon class glutathione transferase confers DDT resistance in the dengue vector, Aedes aegypti.
Glutathione transferases (GSTs) play a central role in the detoxification of xenobiotics such as insecticides and elevated GST expression is an important mechanism of insecticide resistance. In the mosquito, Anopheles gambiae, increased expression of an Epsilon class GST, GSTE2, confers resistance to DDT. We have identified eight GST genes in the dengue vector, Aedes aegypti. Four of these belong to the insect specific GST classes Delta and Epsilon and three are from the more ubiquitously distributed Theta and Sigma classes. The expression levels of the two Epsilon genes, a Theta GST and a previously identified Ae. aegypti GST [Grant and Hammock, 1992. Molecular and General Genetics 234, 169-176] were established for an insecticide susceptible and a resistant strain. We show that the putative ortholog of GSTe2 in Ae. aegypti (AaGSTe2) is over expressed in mosquitoes that are resistant to the insecticides DDT and permethrin. Characterisation of recombinant AaGSTE2-2 confirmed the role of this enzyme in DDT metabolism. In addition, unlike its Anopheles ortholog, AaGSTE2-2 also exhibited glutathione peroxidase activity.
Aedes↗
Expression profiling of a diapause-specific peptide (DSP) of the leaf beetle Gastrophysa atrocyanea and silencing of DSP by double-strand RNA.
A diapause-specific peptide (DSP) composed of 41 amino acid residues including 6 cysteines, has been isolated from diapausing adults of the leaf beetle Gastrophysa atrocyanea. In this study, DSP was found to be expressed primarily in diapausing adults and to a minor extent in pupae, but not in eggs, larvae, or post-diapausing adults. DSP was not induced by bacterial or fungal challenge. DSP-less adults were generated by the injection of double-stranded RNA (dsRNA) corresponding to the dsp gene into pre-diapausing adults. Gene silencing induced by dsRNA was found to be a useful tool for the analysis of DSP in diapausing adults. DSP-less adults showed similar burrowing behavior and oxygen consumption as control insects suggesting that DSP is not essential for the normal onset and maintenance of diapause.
Animals↗
Transfer and fate of seminal fluid molecules in the beetle, Diaprepes abbreviatus: implications for the reproductive biology of a pest species.
Molecules transferred from males to females via seminal fluids are important to the study of insect reproduction because they affect female physiology, reproductive behavior, and longevity. These molecules (seminal fluid molecules or SFMs) interest applied entomologists because of their potential use in insect control. SFMs are also interesting because of their relatively rapid evolution and important role in post-mating sexual selection. We studied SFMs in Diaprepes abbreviatus, a major pest of numerous plant species of economic importance. Using radiolabeled-methionine (35S), we found that D. abbreviatus males synthesized proteins de novo in their reproductive tissues after mating. Males that were fed radiolabeled methionine transferred radioactivity to females beginning within the first 10 min of mating. Male-derived substances are absorbed from the female's reproductive tract into the hemolymph and circulated throughout the body, but are found primarily in the eggs and ovaries. As a result, SFMs may be a useful means of both horizontal (to mates) and vertical transfer (to offspring) of control agents between conspecifics.
Animals↗
Vegetative compatibility groups in indigenous and mass-released strains of the entomopathogenic fungus Beauveria bassiana: likelihood of recombination in the field.
Using nitrate non-utilizing (nit) mutants, we determined vegetative compatibility groups (VCG) among strains of Beauveria bassiana representing strains indigenous to North America, isolated from diverse insect hosts, and strains that have been mass released for insect control. Genetic similarity among these strains was analyzed using random amplified polymorphic DNA (RAPD) markers. Our data revealed 23 VCGs among the 34 strains tested, with most of these groups comprised of only a single strain. We also observed a VCG comprised of eight genetically similar strains isolated from Colorado potato beetles (CPB). Co-inoculation studies of CPB larvae with complementary nit mutants from the same or from different VCGs revealed heterokaryosis in four out of five same-VCG pairs, with only 5-17% of the sporulating cadavers generating few parasexual recombinants. In contrast, none of the infected beetles treated with non-compatible pairs generated recombinants. The large number of VCGs observed and the low frequency of in vivo recombination limited to vegetatively compatible strains indicate that this self/non-self recognition system may be an effective barrier preventing genetic exchange between dissimilar strains in the field.
Animals↗