Natural infection of the tree hole breeding mosquito Aedes triseriatus (Diptera: Culicidae) with the fungus Funicularius triseriatus.
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Aedes↗
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Cyanide induced aversions in the possum (Trichosurus vulpecula): effect of route of administration, dose, and formulation.
Possums (Trichosurus vulpecula) are New Zealand's main vertebrate pest. Control practices using poisons are likely to remain the most cost effective methods for the immediate future but poisons may be avoided in the field. A series of experiments were conducted to determine whether cyanide bait avoidance involved conditioned food aversions (CFA) induced by sublethal cyanide ingestion. Food aversions were conditioned in three experiments using intraperitoneal or oral routes of administration with three different cyanide formulations over a range of doses. Across all experiments there was a direct relationship between dose and the proportion of animals developing CFAs. When administered orally, doses greater than 5 mg/kg resulted in more than 50% of surviving animals developing aversions. Route of administration or formulation appeared to have no differential effect on development of CFA. Although there was considerable variation between individuals in the degree of aversion shown this may be one mechanism that could account for poor possum kill rates in some poisoning operations.
Administration, Oral↗
New plasmid vector and host system for genetic engineering in Bacillus sphaericus.
A new plasmid, pNQ116, was constructed in Bacillus sphaericus by cloning a promoter fragment from B. sphaericus Ts-1 into pNQ112. The plasmid (CmrKmr, 5.23 kb) contains a restriction endonuclease polylinker used for cloning foreign genes, and its cat-86 gene is expressed at high levels from the Ts-1 promoter. This plasmid vector has been transformed into B. sphaericus AS 1.270, AS 1.465, AS 1.469, and 2362, at frequencies of 10(2)-10(3) transformants per microgram of DNA, and is maintained stably under nonselective conditions in these host strains. The presence of pNQ116 in B. sphaericus 2362 does ot interfere with the mosquito larvicidal activity of the organism.
Animals↗
Mutagenicity testing of the juvenoid methoprene (ZR-515) by means of the Drosophila wing spot test.
The juvenile hormone analogue methoprene, which is used in insect pest control, was subjected to mutagenicity testing by means of the Drosophila wing spot test. Larvae heterozygous for recessive wing trichome mutations were exposed to a sublethal dose of methoprene. Wings of emerged adult females were inspected for the presence of phenotypically mutant mosaic spots. Methoprene exhibited a weak mutagenic effect. The fact that only small mosaic clones were induced is discussed.
Animals↗
Genetic safety evaluation of pesticides in different short-term tests.
Cyanazine, cyhexatin, dicamba and DNOC are pesticides commonly and broadly used in agriculture pest control. However, there is little information on their toxicity and mutagenicity in human cells and in whole animals. Therefore, UDS assay and SCE assay in human peripheral lymphocytes, and chromosome aberration analysis in bone marrow of rats have been used to assess the DNA-damaging activity of the above pesticides. Cyanazine proved non-genotoxic in all the test systems. Cyhexatin showed only weakly positive results for SCE induction in human lymphocytes, providing no concern for genotoxicological hazard. While dicamba did not show clastogenic effects in rodents, DNOC gave significant dose-related increases of structural chromosome aberrations in rat bone marrow cells. Female animals showed increased sensitivity to the toxic effects by DNOC at the highest dose. The results provide further information on the intrinsic genotoxic activity of the tested pesticides, which may contribute to the toxicological assessment of the risk associated with human exposure.
Animals↗
Polymerase chain reaction and non-radioactive gene probe based identification of mosquito larvicidal strains of Bacillus sphaericus and monitoring of B. sphaericus 1593M, released in the environment.
In attempts towards an accurate monitoring in the environment, the status of the deliberate release of Bacillus sphaericus, a powerful mosquito larvicidal agent, as well as to evolve a rapid method of screening for potent isolates of B. sphaericus, we have developed a polymerase chain reaction (PCR) method of analysis. Using specific primers spanning the 5' and 3' ends of the coding sequences of these two mosquito larvicidal genes, a 1.3 and a 1.1 kb product from gene A and a 2.6 kb product from gene B have been amplified by PCR. The primers and the products amplified from them in PCR are highly specific for B. sphaericus. We used digoxigenin based non-radioactive chemiluminescence method for the detection of PCR products and the sensitivity of the method was high enough to detect the presence of 1 to 5 cells of B. sphaericus. A simple and inexpensive sample processing procedure has also been developed for direct PCR amplification of B. sphaericus DNA from field samples collected from areas where it had been applied. Several highly toxic to non-toxic strains of B. sphaericus were screened with these primers by PCR for the presence of either or both of these toxin genes. The results indicate that there is a good correlation between the presence of both genes with higher toxicity of the strains.
Animals↗
Residue analysis of DDT, malathion and kelthane on pears.
Malthion, DDT, kelthane and their mixture (3 : 10 : 5) were applied to pear fruits at the rate recommended by the Ministry of Agriculture. The rate of disappearance of their residues on pear fruits was investigated under refrigerated conditions (2 +/- 1 degree and 47% relative humidity (RH)). The same insecticides were tested in higher concentrations (1.5 and 2 times) to determine the effect of insecticides on the black spots usually found on pears after storage. No relation was found between the tested insecticides and the appearance of the black spots. Residue-disappearance curves of the three tested concentrations of these insecticides and their mixture were drawn on semi-log paper. The residues of each material found on pear fruits after one hour, 1, 8, 15, 30 and 45 days' storage in a refrigerator, estimated biologically by C. pipiens larvae and D. magna were recorded. The highest concentrations of the mixture and of the individual insecticides left higher residues on pears than the lower ones of all the insecticides either as a mixture or alone. Stored pears, refrigerated under the above conditions immediately after treatment with the mixture of insecticides recommended for the pest control of pears, cannot be consumed fresh after 45 days.
Animals↗
Synthesis and expression of genes encoding putative insect neuropeptide precursors in tobacco.
Neuropeptides are the key molecules in a multiplicity of physiological processes and their use in pest control has recently been suggested. Most neuropeptides are produced in the form of a precursor that is cleaved by proteolysis to yield various biologically active peptides. To mimic this structure, a method has been developed for synthesizing genes that code for putative polyneuropeptide precursors. As a model neuropeptide, the 5-amino-acid proctolin, one of the best studied invertebrate neuropeptides, functioning both as a visceral and a skeletal neuromuscular transmitter, was chosen. The synthetic gene was introduced into bacteria and tobacco plants, where it was efficiently transcribed. We present our results as a possible approach for the expression, in a variety of organisms, of synthetic genes coding for a wide repertoire of insect neuropeptides.
Amino Acid Sequence↗
Effects of chlordane on parameters of liver and muscle toxicity in man and experimental animals.
An attempt was made to compare the toxic effects of the organochlorine insecticide 'chlordane' in man and rats. Analysis of blood for chlordane metabolites showed their presence in the descending order of trans-nonachlor, oxychlordane, heptachlorepoxide and cis-nonachlor. The total range of chlordane and its metabolites in the sera of workers was 9.84 +/- 4.47 ng/g. Serum levels of triglycerides (TG), creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) activities were also found to be higher in pest-control operators. In a simultaneous study, rats were administered 100 mg/kg body wt. of chlordane by stomach tube once a day for 4 days, whereas 50 mg/kg body wt. of chlordane was injected intraperitoneally once a day for 4 days. The data show that total cholesterol and serum TG as well as CPK and LDH activities are increased after chlordane treatment. The isoenzyme patterns suggest that an increase in CPK and LDH is related to skeletal muscle. Furthermore, the hepatotoxicity of chlordane was also studied in rats only. A significant increase in liver weight, its water content, total lipids, triglycerides and phospholipids was recorded. Chlordane induced lipid peroxidation in the liver, exhibiting a dose-response relationship. Although no appreciable effect on mitochondrial function and latent ATPase activity was observed, 2,4-dinitrophenol-stimulated ATPase activity was inhibited. Histological examination of the liver confirmed fatty infiltration induced by chlordane in rats.
Adolescent↗
Confirmation method for the identification and determination of some organophosphorus and organochlorine pesticides in cocoa beans by gas chromatography-mass spectrometry.
The main objective of this work is to develop a routine quality control method for pesticide residues in cocoa beans, using gas chromatography-mass spectrometry. The investigated pesticides, which are used to control pests in the growing of cacao, are: Acephate, Propoxur, HCH, Heptachlor, Fenitrothion, Pirimiphos-methyl, Aldrin, Dieldrin, pp'-DDE, op-DDE and DDT. Two extraction methods were tested. The first was based on strong attack by concentrated sulphuric acid and later extraction with n-hexane: the investigated residues were Acephate, HCH, Fenitrothion and DDT; recoveries were 68-95% and the detection limits 0.5-10 ppb. The second extraction method was based on the Universal Trace Residue Extractor (UNITREX), which consists of a distillation system for organophosphorus and organochlorine pesticides in fatty samples. The investigated residues were Heptachlor, Pirimiphos-methyl, Aldrin, Propoxur, Dieldrin, op-DDE and pp'-DDE; recoveries were 67-88% and the detection limits 1-10 ppb.
Cacao↗
Neonatal exposures to technical methoxychlor alters ovaries in adult mice.
Technical grade methoxychlor (MXC) is an estrogenic pesticide currently used for pest control in the US. To determine the long-term effects of technical MXC on ovaries and estrous cycles, neonatal mice received 14 daily intraperitoneal injections of sesame oil only, 10.0 micrograms estradiol-17 beta, or 0.05 mg, 0.1 mg, 0.5 mg, or 1.0 mg of technical MXC. At 3, 6, and 12 months, vaginal smears were examined for 12 d and ovaries collected. All technical MXC doses and estradiol increased the duration of vaginal cornification but only estradiol, 0.5, or 1.0 mg technical MXC induced ovarian atrophy, relative ovarian weight loss, and depletion of corpora lutea. Technical MXC doses of 0.05 or 0.1 mg produced the opposite effects; the ovaries remained heavy, large, and filled with corpora lutea. On the other hand, follicular cysts were recorded in all groups, except the 1.0 mg MXC group. These augmentary and inhibitory effects of MXC at low and high doses mimic the estrogen effects at low and high doses, and were probably due to the altered hypothalamic-hypophyseal function.
Animals↗
Soil and groundwater transport of microorganisms.
Releases of GEMs into the environment are expected to increase in the next few years, with the most dramatic increases resulting from the application of pest-control agents in agriculture and forestry. Of major significance in assessing the environmental risk impact of GEMs is an understanding of their survival and transport in soil and subsurface environments. While information on the transport and survival of microorganisms through soil is available, it is neither abundant nor extensive in terms of microbial types tested or soils examined. Though the transport of microorganisms from an application site depends primarily upon passive mechanisms, broad generalizations pertaining to the transportability of a specific microorganism within a particular soil environment may not be possible. Indeed, to extrapolate from information about one microbe to another, or from one geographical location to another, may not be appropriate. What is clear, however, is that the broader the data base, the more powerful the argument for making reasoned judgement, and consequently the more satisfactory the results of the predictive process. Several inherent difficulties exist in studying the transport behavior of GEMs in soil and subsurface environments. Detection of low microbial numbers or of stressed microbial populations is exceedingly difficult with traditional technology. In an effort to improve detection sensitivity, many improved methods of monitoring GEMs in the soil and subsurface are currently under development (Chaudhry et al. 1989). Beyond the difficulties of making accurate measurements of microorganisms to determine their spatial and temporal situation in the soil and subsurface environment, lies the need to ascertain the dynamic relationships between indigenous populations of microorganisms and how they may interact with a released GEM. Also, research strategies have not adequately addressed methods to predict the potential interactions between GEMs and natural microorganisms. These issues must be addressed if environmental risk assessment is to be valuable. There is clearly a need for focused research on the survival and transport of GEMs in these environments.
Bacteria↗
Enantiospecific (+)- and (-)-germacrene D synthases, cloned from goldenrod, reveal a functionally active variant of the universal isoprenoid-biosynthesis aspartate-rich motif.
The naturally occurring, volatile sesquiterpene hydrocarbon germacrene D has strong effects on insect behaviour and genes encoding enzymes that produce this compound are of interest in the study of plant-insect interactions and in a number of biotechnological approaches to pest control. Goldenrod, Solidago canadensis, is unusual in that it produces both enantiomers of germacrene D. Two new sesquiterpene synthase cDNAs, designated Sc11 and Sc19, have been isolated from goldenrod and functional expression in Escherichia coli identified Sc11 as (+)-germacrene D synthase and Sc19 as (-)-germacrene D synthase. Thus, the enantiomers of germacrene D are the products of separate, but closely related (85% amino-acid identity), enzymes. Unlike other sesquiterpene synthases and the related monoterpene synthases and prenyl transferases, which contain the characteristic amino-acid motif DDXX(D,E), Sc11 is unusual in that this motif occurs as (303)NDTYD. Mutagenesis of this motif to (303)DDTYD gave rise to an enzyme that fully retained (+)-germacrene D synthase activity. The converse mutation in Sc19 (D303N) resulted in a less efficient but functional enzyme. Mutagenesis of position 303 to glutamate in both enzymes resulted in loss of activity. These results indicate that the magnesium ion-binding role of the first aspartate in the DDXXD motif may not be as critical as previously thought. Further amino-acid sequence comparisons and molecular modelling of the enzyme structures revealed that very subtle changes to the active site of this family of enzymes are required to alter the reaction pathway to form, in this case, different enantiomers from the same enzyme-bound carbocationic intermediate.
Alkyl and Aryl Transferases↗
Inhibition of mevalonate 5-diphosphate decarboxylase by fluorinated substrate analogs.
Mevalonate 5-diphosphate decarboxylase (MDD) is a peroxisomal enzyme in the cholesterol biosynthetic pathway, which plays an important role in regulating cholesterol biosynthesis. In the present study, rat MDD was cloned and purified to apparent homogeneity. Two fluorinated MDD substrate analogs, P'-geranyl 2-fluoromevalonate 5-diphosphate (4) and 2-fluoromevalonate 5-diphosphate (6), were synthesized, and both were found to be irreversible inhibitors of rat MDD. These two inhibitors were characterized, and mechanisms of the inactivation process were proposed. Kinetic studies indicate both analogs only bind into mevalonate binding-site of MDD. Compound 4 shows competitive inhibition on mevalonate kinase (MVK), and its IC(50) value was determined to be comparable with that of geranyl diphosphate. Further kinetic studies indicate compound 4 only bind into ATP binding-site of MVK. These studies provide an example for a single inhibitor to carry out sequential blocking of two enzymes in cholesterol biosynthesis, which may provide useful information for drug discovery for the purpose of treating cardiovascular disease and cancer or for pest control.
Animals↗
Interaction between organophosphate compounds and cholinergic functions during development.
Organophosphate (OP) compounds exert inhibition on cholinesterase (ChE) activity by irreversibly binding to the catalytic site of the enzymes. For this reason, they are employed as insecticides for agricultural, gardening and indoor pest control. The biological function of the ChE enzymes is well known and has been studied since the beginning of the XXth century; in particular, acetylcholinesterase (AChE, E.C. 3.1.1.7) is an enzyme playing a key role in the modulation of neuromuscular impulse transmission. However, in the past decades, there has been increasing interest concerning its role in regulating non-neuromuscular cell-to-cell interactions mediated by electrical events, such as intracellular ion concentration changes, as the ones occurring during gamete interaction and embryonic development. An understanding of the mechanisms of the cholinergic regulation of these events can help us foresee the possible impact on environmental and human health, including gamete efficiency and possible teratogenic effects on different models, and help elucidate the extent to which OP exposure may affect human health. The chosen organophosphates were the ones mainly used in Europe: diazinon, chlorpyriphos, malathion, and phentoate, all of them belonging to the thionophosphate chemical class. This research has focused on the comparison between the effects of exposure on the developing embryos at different stages, identifying biomarkers and determining potential risk factors for sensitive subpopulations. The effects of OP oxonisation were not taken into account at this level, because embryonic responses were directly correlated to the changes of AChE activity, as determined by histochemical localisation and biochemical measurements. The identified biomarkers of effect for in vitro experiments were: cell proliferation/apoptosis as well as cell differentiation. For in vivo experiments, the endpoints were: developmental speed, size and shape of pre-gastrula embryos; developmental anomalies on neural tube, head, eye, heart. In all these events, we had evidence that the effects are mediated by ion channel activation, through the activation/inactivation of acetylcholine receptors (AChRs).
Animals↗
The modulatory effect of deltamethrin on antioxidants in mice.
BACKGROUND: Deltamethrin is a alpha-cyano pyrethroid insecticide used extensively in pest control. Although initially thought to be least toxic, a number of recent reports showed its toxicity in mammalian and non-mammalian laboratory and wildlife animal species. In fish, it is a potent oxidative stress-inducing agent. We studied the oxidative stress-inducing effect of deltamethrin in mice. METHODS: Male Swiss albino mice were orally administered 2 doses of deltamethrin viz., 5.6 and 18mg/kg body weight (bw), for 15 days. RESULTS: Both the doses of deltamethrin significantly induced lipid peroxidation (LPO) in liver and kidney. Along with the induction of LPO, activities of vital antioxidant enzymes such as glutathione peroxidase (GPx), glutathione S-transferase (GST) and catalase (CAT) were also suppressed in both the tissues. Glutathione (GSH) level was also decreased. GSH decrease was more pronounced in kidney than the liver. CONCLUSION: Toxicity of many chlorinated and organophosphate insecticides is mediated by the reactive oxygen species (ROS). Findings of the present investigation also suggest a role for ROS in deltamethrin toxicity. An increased LPO indicates that these ROS might have caused degradation of biomembrane in deltamethrin-exposed animals.
Animals↗
Acute toxicity of alpha-cypermethrin to guppy (Poecilia reticulata, Pallas, 1859).
Alpha-cypermethrin is a synthetic pyrethroid insecticide used to control pests in domestic, industrial and agricultural situations. Adult male guppy fish (Poecilia reticulata standard test species) were selected for the bioassay experiments. The experiments were performed in four series and the 96-h LC50 value was determined for guppies. The acute toxicity experiments were carried out by static method and behavioral changes in guppies were determined for each alpha-cypermethrin (98% technical grade) concentration. Data obtained from the alpha-cypermethrin investigation were evaluated by the use of probit analysis statistical method and the 96-h LC50 value for guppy was estimated as 9.43 microg/l.
Animals↗
Chlorpyrifos-methyl dissipation in a small adjacent water body following application to citrus.
Chlorpyrifos-methyl use in citrus, vineyards and vegetables is extensive and it has become an essential component of pest control in Southern European regions. The objective of this study was to assess surface water and sediment exposure to chlorpyrifos-methyl (RELDAN 22) under field conditions in a citrus orchard in Sicily (Italy). Pesticide drift loadings were measured in a small surface water body situated 0.5-1.0 m from the edge of the treated field. Measured drift values after the application were between 0.04% and 0.19% of the theoretical applied dose and were highly variable. However, the loadings were lower than those measured by other authors, also lower than predicted by regulatory drift models (95th percentile) typically used for aquatic risk assessment in Europe. Most of the variability occurred due to fluctuating wind speed and direction. In water samples collected in the surface water adjacent to the target field, chlorpyrifos-methyl was detected immediately after application, with concentrations ranging from less than the limit of quantification of the analytical method (0.05 microg/l) to a maximum of 0.08 microg/l. Predicted environmental concentrations in water, using the TOXSWA model, were similar to the measured data when measured drift data from the field experiment were used as inputs.
Chlorpyrifos↗