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Seasonal variation of persistent organochlorine insecticide residues in Vellar River waters in Tamil Nadu, South India.

Water samples collected from Vellar river and Pichavaran mangroves at Porto Novo (11 degrees 29' N, 79 degrees 46' E), Tamil Nadu State, South India, from December 1987 to January 1989 were analyzed to determine the seasonal variation of the levels of organochlorine insecticides such as HCH (BHC) and DDT. Both these insecticides showed higher levels from October to February, although this trend was more pronounced in HCH than DDT, reflecting the application of technical HCH largely and probably small quantities of DDT during the flowering season of rice. The alpha-HCH was detected as a dominant isomer for all seasons monitored followed by beta-HCH. Among DDT compounds, p,p'-DDT was the highest in river water except in the dry season when p,p'-DDD showed a higher percentage. On the other hand, in mangroves p,p'-DDE was highest during the wet season and p,p'-DDD during the dry season. Air-water partitioning data of HCH isomers and DDT compounds in Vellar river revealed that these chemicals tend to be in the water phase. These observations may aid in understanding the role of a tropical paddy area on the behavior and fate of man-made chemicals in view of worldwide contamination.

Journal Article↗

Buffer zone widths for honeybees from ground and aerial spraying of insecticides.

The relative field hazards of insecticides to honeybees have been estimated by considering intrinsic toxicity levels and field application rates. This approach is extended here to a consideration of buffer zones downwind of sprayed areas by estimating the distance at which bees would encounter an LD(50) dose from spray drift. 'LD(50) distances' are determined for both ground and aerial spraying of ground crops in Britain using published data on spray deposition under various weather conditions. For ground spraying at low wind speeds (< or =3 m s(-1)), this zone of risk is up to 5 m for the great majority of compounds. Aerial spraying in unstable atmospheric conditions appears to produce drift deposits of about the same order of magnitude as from ground spraying at wind speeds of about 4 m s(-1), with maximum LD(50) distances of < or =40 m for chlorpyrifos, fenitrothion and triazophos. For aerial spraying in stable atmospheric conditions these distances would be much greater. Pieris brassicae larvae are contrasted with honeybees in their relative sensitivities to insecticides and consequent LD(50) distances.

Journal Article↗

Dissipation of some organochlorine insecticides in cropped and uncropped soil.

Dissipation of four organochlorine insecticides, viz. aldrin, HCH, chlordane and heptachlor was studied in a sandy loam soil with and without crops during a period of 10 cropping seasons. Dissipation of all chemicals followed first-order kinetics (r(2)=0.537 - 0.976) with almost similar persistence in cropped and uncropped soils for all the insecticides. The average half-lives, (t(1/2) values) for total residues of aldrin, HCH, chlordane, and heptachlor in cropped treatments were 80.7, 58.8, 93.2, and 110 days. Their respective values in fallow plots were 78.4, 83.8, 154, and 116 days. None of the parent compounds or their isomers could be detected below the 20 cm depth at the termination of the experiment. Highest residue concentrations were observed in the surface 10 cm layer in fallow plots, but in the deeper (10-20 cm) layer in cropped plots. Analysis of plants and grains showed significant residues of all the chemicals. Degradation of these compounds in cropped and uncropped plots is discussed with regard to their volatilization, microbial degradation, leaching, and plant uptake.

Journal Article↗

Polychlorinated biphenyl and organochlorine insecticide residues in human adipose tissue in Poland.

Concentrations of PCBs (polychlorinated biphenyls) and organochlorine insecticides such as DDT, HCHs (hexachlorocyclohexanes), HCB (hexachlorobenzene) and chlordanes were determined in human adipose fat from the provinces of Skierniewice and Gdańsk in Poland collected during 1979 and 1990, respectively. The mean levels (microg g(-1) lipid weight basis) in Skierniewice and Gdańsk were found to be 1.2+/-0.44 and 1.5+/-1.3 for PCBs, 25+/-16 and 15+/-13 for DDT, 0.53+/-0.17 and 0.25+/-0.09 for HCHs, 0.36+/-0.22 and 0.26+/-0.23 for HCB, and 0.11+/-0.08 and 0.07+/-0.04 for chlordanes, respectively. Very high concentrations of DDT were detected, with the maximum value as high as 52 microg g(-1) lipid weight in Skierniewice and 47 microg g(-1) lipid weight in Gdańsk. PCB residues contained in the human fat from Polish cadavers are comparable to those of the levels reported for industrialized nations. It is appraised from the levels of DDT and PCBs in Poles' fat that the Polish environment is still experiencing both types of contamination, arising from agricultural and industrial activities. Contamination of humans by other organochlorine insecticides was relatively lower.

Journal Article↗

Metabolic activation of the organophosphorus insecticides chlorpyrifos and fenitrothion by perfused rat liver.

The present study was undertaken to characterize the metabolic activation of the organophosphorus insecticides chlorpyrifos [O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothionate] and fenitrothion [O,O-dimethyl O-(3-methyl-p-nitrophenyl) phosphorothionate] by intact rat liver. Single-pass perfusions of rat livers with chlorpyrifos or fenitrothion to steady state conditions resulted in the appearance of their corresponding oxygen analogs in effluent. In addition, detoxification of chlorpyrifos oxon [O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphate] or fenitrooxon [O,O-dimethyl O-(3-methyl-p-nitrophenyl) phosphate] by rat blood did not proceed at a rate rapid enough to prevent passage of at least some of these chemicals from liver to extrahepatic tissues, suggesting that hepatic biotransformation of chlorpyrifos and fenitrothion by rat liver results in their net activation. Although male rat livers produced more chlorpyrifos oxon and fenitrooxon from chlorpyrifos and fenitrothion, respectively, than did livers from female rats, the acute toxicities of chlorpyrifos and fenitrothion were greater in females than in males. Therefore, differences in hepatic activation of chlorpyrifos and fenitrothion in males and females cannot account for the sex differences in their acute toxicities in the rat. Finally, S-methyl glutathione and S-p-nitrophenyl glutathione were not detected in effluent or bile of livers perfused with fenitrothion, suggesting that glutathione-mediated biotransformation of this insecticide does not occur to any significant degree in intact liver.

Animals↗

Nucleotide sequence of the insecticidal protein gene of Bacillus thuringiensis strain aizawai IPL7 and its high-level expression in Escherichia coli.

A DNA fragment carrying the insecticidal protein gene of Bacillus thuringiensis subsp. aizawai IPL7 was cloned from a 78-kb plasmid. The nucleotide sequence revealed that the cloned DNA fragment contained a 3465-bp protein-coding region with 156-bp 5'-flanking, and 168-bp 3'-flanking regions. The open reading frame encoded a 130,690 Da protein consisting of 1155 amino acid residues. Nucleotide sequence comparison of the aizawai gene with the published berliner 1715 gene showed only 8 nt changes in the coding regions. It was found that 72 bp of the 5'-flanking sequence of the cloned aizawai gene was responsible for constitutive expression of the 130-kDa protein gene in Escherichia coli. The expression was greatly enhanced by introducing the tac promoter upstream from the 72-bp 5'-flanking region of the aizawai gene. Under optimal conditions, the 130-kDa insecticidal protein amounted to 38% of the total cellular protein.

Amino Acid Sequence↗

Cytological effects of the insecticide tetrachlorvinphos.

The effect of the organophosphorus insecticide tetrachlorvinphos (Gardona) has been studied on the mitosis and meiosis of Vicia faba, using the pure insecticide. An aqueous saturated solution of tetrachlorvinphos affected neither mitosis (after seed-soak and root treatment) nor meiosis (after spraying the plants at the flowering stage). The obtained results may be attributed to the low solubility of tetrachlorvinphos. A saturated solution of tetrachlorvinphos in Tween-60: water mixture (1:99) induced a statistically significant percentage of abnormal cells in root-tip meristems after root treatment for 4 h. Chromosome stickiness, disturbed meta- and anaphases and anaphase bridges were observed.

Meiosis↗

Biotransformation of the insecticide parathion by mouse brain.

The acute toxicity of organothiophosphate insecticides like parathion results from their metabolic activation by cytochromes P450. The present study is directed towards the characterization of cytochrome-P450-dependent metabolism of parathion by various mouse brain regions. Intraperitoneal administration of [35S]parathion to mice led to covalently bound [35S]sulfur in various tissues, indicating their capacity to oxidatively desulfurate this insecticide. Liver contained the greatest amount of covalently bound sulfur, and brain the least. Among individual brain regions the olfactory bulb and hypothalamus possessed the highest levels of sulfur binding when expressed on a per milligram tissue basis. However, when expressed on a per brain region basis, sulfur binding was greatest within the cortex as a result of the large mass of this region, compared to the hypothalamus and olfactory bulb. Incubation of the 78,000 x g fraction of mouse brain with parathion resulted in formation of p-nitrophenol, although paraoxon could not be detected. However, given the current understanding of parathion metabolism by cytochromes P450, and given that paraoxon can rapidly disappear through phosphorylation of serine hydroxyl groups, it is reasonable to assume that at least some paraoxon was formed. Production of p-nitrophenol required NADPH and was inhibited by carbon monoxide. In vitro incubations of parathion with the 78,000 x g fraction of mouse brain indicated that the hypothalamus and olfactory bulb had the greatest capacity to produce p-nitrophenol. These results demonstrate that various mouse brain regions possess different capacities to metabolize parathion.

Animals↗

Effects of pyrethroid insecticides on operant responding maintained by food.

The pyrethroids are potent insecticides with low concomitant mammalian lethality when compared with other major insecticides. While high doses can lead to hyperactivity, tremors, convulsion and death, low doses have not been as well studied. Since operant behavior can be a sensitive measure of CNS function, male Holtzman rats were trained on a VR25 schedule maintained by 45 mg food pellets. Rats were injected IP with one of four different technical grade pyrethroids: permethrin, allethrin, deltamethrin and fenvalerate. All agents were effective in reducing operant responding and did so in a dose-dependent manner at levels 10 to 100 times below their LD50 values. Time course studies indicated a relatively short duration of action for the Type I agents of less than 60 min for permethrin and 15 min for allethrin. Type II agents were generally effective for greater than 60 min. Results of these studies indicate that operant responding maintained by food is a sensitive measure of the behaviorally disruptive effects of subconvulsive doses of pyrethroids.

Allethrins↗

Cholin- and carboxylesterase activities in developing zebrafish embryos (Danio rerio) and their potential use for insecticide hazard assessment.

Insecticides are a potential hazard for non-target organisms like fish particularly at run off events. The study of effects to embryos of the zebra fish Danio rerio is already an accepted tool in waste water monitoring, but effects of various groups of substances (like most pesticides) to the zebrafish embryo remain to be studied. Enzymes are often taken as biomarkers of exposure and effect. Therefore cholinesterase isozymes and carboxylesterase were examined for their suitability as biomarkers of insecticide exposure. The activities of cholinesterase and of carboxylesterase were monitored in the first 48 h post-fertilization (hpf) of zebrafish development. Significant specific activities in the range of 0.5-25 U could be measured from the sixth somite stage (12 h) up to the Long Pec stage (48 h) for different cholinesterases using acetyl-, acetyl-beta-methyl-, butyryl- and propionylthiocholin as substrates. The specific activity of carboxylesterase ranged from 4 to 16 Umg(-1) protein in the respective developmental stages. Substrate specificity was analysed using specific inhibitors (eserine sulphate, DPDA, BW284c51). The results showed that the observed cholinesterase activities in the whole embryo may be attributed mainly to acetylcholinesterase with a partial capability to use propionylthiocholine as a second substrate. The potential use of cholin- and carboxylesterase as biomarkers was investigated using the organophosphate paraoxon-methyl. A 40% inhibition of enzyme activities was reached by 0.4 microM paraoxon-methyl indicating the possible use of these enzymes as biomarkers of exposure.

Animals↗

Simple and rapid assay for analyzing residues of carbamate insecticides in bovine milk: hot water extraction followed by liquid chromatography-mass spectrometry.

A simple, specific and rapid procedure for determining six largely used carbamate insecticides in bovine whole milk is here presented. This method is based on the matrix solid-phase dispersion technique with heated water as extractant followed by liquid chromatography (LC)-mass spectrometry (MS) equipped with a single quadrupole and an electrospray ion source. Target compounds were extracted from milk by water heated at 90 degrees C. After acidification and filtration, 0.2 mL of the aqueous extract was injected in the LC column. MS data acquisition was performed in the selected ion-monitoring mode, selecting three ions for each target compound. Heated water appeared to be an excellent extractant, since absolute recovery data ranged between 76 and 104% with R.S.D. not larger than 8%. Using butocarboxim (an obsolete carbamate insecticide) as surrogate internal standard, the accuracy of the analysis at three spike levels varied between 85 and 105% with R.S.D. not larger than 9%. On the basis of a signal-to-noise ratio of 10, limits of quantification were estimated to range between 3 ppb (propoxur) and 8 ppb (pirimicarb). The effects of temperature, volume and flow rate of the extractant on the analyte recovery were studied.

Animals↗

Isolation, characterization and expression of a novel vegetative insecticidal protein gene of Bacillus thuringiensis.

Twenty-four serovars of Bacillus thuringiensis (Bt) were screened by polymerase chain reaction to detect the presence of vegetative insecticidal protein gene (vip)-like sequences by using vip3Aa1-specific primers. vip-like gene sequences were identified in eight serovars. These genes were cloned and sequenced. The deduced amino acid sequence of the vip3Aa14 gene from Bacillus thuringiensis tolworthi showed considerable differences as compared to those of Vips reported so far. The vip3Aa14 gene from Bt tolwarthi was expressed in Escherichia coli using expression vector pET29a. The expressed Vip3Aa14 protein was found in cytosolic supernatant as well as pellet fraction, but the protein was more abundant in the cytosolic supernatant fraction. Both full-length and truncated (devoid of signal sequence) Vips were highly toxic to the larvae of Spodoptera litura and Plutella xylostella. Truncation of Vip3Aa14 protein at N-terminus did not affect its insecticidal activity.

Animals↗

The molecular basis of insecticide resistance in mosquitoes.

Insecticide resistance is an inherited characteristic involving changes in one or more insect gene. The molecular basis of these changes are only now being fully determined, aided by the availability of the Drosophila melanogaster and Anopheles gambiae genome sequences. This paper reviews what is currently known about insecticide resistance conferred by metabolic or target site changes in mosquitoes.

Animals↗

Isolation, geographical diversity and insecticidal activity of Bacillus thuringiensis from soils in Spain.

Bacillus thuringiensis is a spore-forming bacterium showing the unusual ability to produce endogenous crystals during sporulation that are toxic for some pest insects. This work was performed to study the composition, ecological distribution and insecticidal activity of isolates of this entomopathogenic bacterium from the Spanish territory. Using a standard isolation method, B. thuringiensis was isolated from 115 out of 493 soil samples collected in the Iberian Peninsula and the Canary and Balearic Archipelagos. The percentages of samples with B. thuringiensis were 31.7, 27.6 and 18.5 and the B. thuringiensis index 0.065, 0.067 and 0.11 for the Iberian Peninsula, Canary and Balearic Archipelagos, respectively. The prairies were shown to be the worst source of B. thuringiensis while forests, urban and agricultural habitats showed similar percentages. Strain classification based on H-antigen agglutination showed a great diversity among the Spanish isolates, which were distributed among 24 subspecies, including three new ones andaluciensis, asturiensis and palmanyolensis. We differentiated 65 different protein profiles of spore-crystal mixtures by sulfate-polyacrylamide gel electrophoresis and we selected 109 isolates representative of these profiles to evaluate their insecticidal activity against insects from the Orders Orthoptera, Dictyoptera, Coleoptera, Lepidoptera and Diptera. We found variable percentages of isolates active against Coleoptera and Lepidoptera, one isolate highly active against mosquito larvae and for the first time, three isolates active against cockroaches and locusts.

Animals↗

The plasmid pBMBt1 from Bacillus thuringiensis subsp. darmstadiensis (INTA Mo14-4) replicates by the rolling-circle mechanism and encodes a novel insecticidal crystal protein-like gene.

This work describes a novel rolling-circle replicating (RCR) plasmid pBMBt1 from Bacillus thuringiensis subsp. darmstadiensis (INTA Mo14-4) encoding an insecticidal crystal protein-like gene. pBMBt1 (6700 bp) contains three ORFs and their putative transcription initiation sites and Shine-Dalgarno sequences were localized. ORF1 encodes a 34.6 kDa protein which showed identity with the protein CryC53 from B. thuringiensis subsp. cameroun (24.6%), the Cry15Aa insecticidal crystal protein from B. thuringiensis subsp. thompsoni (21.9%) and the Mtx3 protein from Bacillus sphaericus (27.8%). The ORF2 (52.3 kDa) showed a 74% identity with the Mob protein coded by pUIBI-1 from B. thuringiensis subsp. entomocidus and 64% identity with the Mob protein of pBMY1 from Bacillus mycoides; both Mob proteins belong to the pMV158 superfamily. To evaluate the Mob protein, the plasmid pHTMob14-4 was constructed. This plasmid shows transfer frequencies of 9.1x10(-6) in B. thuringiensis subsp. israelensis (4Q7Gm(R)). The ORF3 (23.6 kDa) gene product is homologous to the Rep protein from the plasmid pBMYdx of B. mycoides (37.6%). A putative double-strand origin with significant homology to that of B. thuringiensis plasmids, and an ssoA-type single-strand origin were also identified. Detection of single-stranded pBMBt1 DNA replicating intermediaries suggests that replication occurs via the rolling-circle mechanism.

Amino Acid Sequence↗

Identification and molecular cloning of insecticidal toxins from the venom of the brown spider Loxosceles intermedia.

The venom of Loxosceles intermedia was investigated for the presence of insecticidal toxins active against Spodoptera frugiperda (Lepdoptera: Noctuidade), an insect that has caused great reductions in corn production in Brazil. A combination of gel filtration (Sephadex G-100) and ion-exchange chromatography (Carboxymethyl Cellulose, CM 52) resulted in four major fractions that were submitted to biological assay. Fraction 4 was further purified by a reverse phase HPLC (C18 Column) resulting in peptides active against Spodoptera frugiperda. Three new potential insecticidal toxins named LiTx x 1, LiTx x 2 and LiTx x 3 were identified. The partial amino terminal sequences of these peptides were obtained and used to clone the corresponding cDNAs with the help of degenerate oligonucleotides. The amino acid sequence deduced from the cDNA of LiTx x 1, LiTx x 2 and LiTx x 3 revealed mature proteins of approximately 7.4, 7.9 and 5.6 kDa.

Amino Acid Sequence↗

Humoral responses to defined malaria antigens in children living since birth under insecticide treated curtains in Burkina Faso.

Insecticide treated materials (ITM) are considered a useful malaria control measure for endemic countries, but whether they also delay the acquisition of immunity to malaria remains unclear. This study investigates plasma antibody levels in 160 children aged 3-6 years from five villages protected by insecticide treated curtains (ITC) over 6 years and in 184 children of the same age group from five villages in the same area never covered by ITC. The antigens to which antibodies were investigated were: the Plasmodium falciparum circumsporozoite protein (CSP) repetitive sequence (NANP)5; the C-terminal domain of the P. falciparum exported protein 1 (Cter-PfExp1); three fragments of the glutamate rich protein (GLURP), referred to as R0, R1 and R2; the merozoite surface protein 3 (MSP3). The level of antibodies was lower in children from the ITC area than in children from the non-ITC area for (NANP)5, R0, R2 and MSP3. Prevalence and intensity of P. falciparum infection were similar in the two groups of children. These findings suggest that reducing the level of malaria transmission over a long period may affect the level of antibodies in children to both sporozoite and blood stage malaria antigens.

Animals↗

Extraction and liquid-solid chromatography cleanup procedures for the direct analysis of four pyrethroid insecticides in crops by gas--liquid chromatography.

The extraction of four pyrethroid insecticides, fenpropanate (WL 41706, S-3206), permethrin (WL 43479, FMC-33297, PP 557, NRDC-143), cypermethrin (WL 43467, NRDC 149), and fenvalerate (WL 43775, S-5602) from carrots, tomatoes celery, and onions with acetone followed by extraction of the resulting acetone--water mixture with hexane, was investigated and found suitable for residue analysis. The elution of the four materials from two types of silica and alumina absorbents with mixtures of organic solvents of varying polarity was examined. The elution patterns and recoveries from silica gel, aluminum oxide, and water deactivated Florisil indicated potential utility in cleanup. Florisil was found to provide cleanup for asparagus, carrots, tomatoes, tobacco, and onions permitting analyses to less than 0.01 ppm for most of the insecticides by electron capture gas--liquid chromatography, at extract concentrations of 1 g/ml. The other adsorbents, singly or in combination, were found to provide the cleanup required for radishes and those cases involving other crops where Florisil cleanup was insufficient.

Chromatography, Gas↗