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Genotoxicity evaluation of pesticide formulations containing alachlor and atrazine in multiple mouse tissues (blood, kidney, liver, bone marrow, spleen) by comet assay.

Every year, in the European countries more than 2 million tons of pesticides are released into the environment. More than 60% of those substances appear to be herbicides. Due to extensive production and application of this chemical their putative detrimental effect on life should be known and minimized. In this study we applied the comet assay on blood and 4 mouse organs (kidney, liver, bone marrow, and spleen) to evaluate possible genome damage caused by two pesticide formulations (Bravo and Gesaprim) containing alachlor and atrazine as active ingredients. Five male CBA mice were assigned to each of 4 treatment groups and control group. Bravo and Gesaprim were injected intraperitoneally once. Two different doses of Bravo were used: 0.031 ml/kg and 0.021 microl/kg, so that doses of alachlor mice received within the pesticide formulation given were 15 mg/kg and 0.01 mg/kg. Also Gesaprim was given at two different doses: 1.08 ml/kg and 0.07 microl/kg so that the doses of atrazine contained within the pesticide formulation given were 540 mg/kg and 3.5 x 10(-2) mg/kg. Mice were sacrificed 24 hours after treatment. Alkaline comet assay on the blood samples, kidney, liver, bone marrow and spleen was performed. Statistically significant (p<0.01) increase of tail length for all 5 tissues examined in mice treated with both Bravo and Gesaprim compared to the control was found. For both pesticides DNA of kidney and liver showed largest increase in migration. Also, distribution of tail length values for Bravo and Gesaprim for all mouse tissues examined showed a shift to the right when compared to the controls.

Acetamides↗

[Construction of multifunctional genetically engineered pesticides-degrading bacteria by homologous recombination].

Construction of multifunctional pesticides-degrading genetically engineered microorganisms (GEMs) is increasing important in the bioremediation of various pesticides contaminants in environment. However, construction of genetically stable GEMs without any exogenous antibiotic resistance is thought to be one of the bottlenecks in GEMs construction. In this article, homologous recombination vectors with the recipient's 16S rDNA as homologous recombination directing sequence (HRDS) and sacB gene as double crossover recombinants positive selective marker were firstly constructed. The methyl parathion hydroalse gene (mpd) was inserted into the 16S rDNA site of the carbofuran degrading strain Sphingomonas sp. CDS-1 by homologous recombination single crossover in the level of about 3.7 x 10-(7) - 6.8 x 10(-7). Multifunctional pesticides-degrading GEMs with one or two mpd genes inserted into the chromosome without any antibiotic marker were successfully constructed. The homologous recombination events were confirmed by PCR and southern blot methods. The obtained GEMs were genetically stable and could degrade methyl parathion and carbofuran simultaneously. The insertion of mpd gene into rrn site did not have any significant effect on recipient' s physiological and original degrading characteristics. The methyl parathion hydrolase (MPH) was expressed at a relatively high level in the recombinants and the recombinant MPH specific activity in cell lysate was higher than that of original bacterium (DLL-1) in every growth phase tested. The highest recombinant MPH specific activity was 6.22 mu/tg. In this article, we describe a first attempt to use rRNA-encoding regions of Sphingomonas strains as target site for expression of exogenous MPH, and constructed multifunctional pesticides degrading GEMs, which are genetically stable and promising for developing bioremediation strategies for the decontamination of pesticides polluted soils.

Biodegradation, Environmental↗

Protective role of ascorbic acid and vitamin B-complex against pesticide-induced clastogeny in bone marrow cells of mice.

Concurrent administration for one week of vitamins B-Complex (0.3 ml of 1% Polybion) or ascorbic acid (0.25 ml of 1% Redoxon, to 8-week old albino swiss mice, Mus musculus along with organophosphorous pesticide Malathion or carbamate pesticide Rogor (both 1 microliters/kg b.wt.) could significantly decrease the chromosome-clastogeny rate induced by the pesticides alone in the metaphase chromosomes of bonemarrow cells. The vitamins were not clastogenic, whereas the Malathion proved to be clastogenic four-times and Rogor six-times more than the control rate. The vitamins seem to reduce the incidence of fragmentation and subsequent rearrangements induced by the pesticides. On the basis of the available literature, it is presumed that the vitamins might be preventing the formation of mutagenic lipid conjugates of biometabolised forms of the pesticides.

Animals↗

[Various principles of the formation of toxic gaseous pesticide degradation products in the soil].

The intensity of gaseous products of pesticides degradation in soil depends on the natural and antropogenic factors: type of soil, hydrothermal conditions, dose of the pesticide, presence in soil of other agrochemicals which can inhibit and also potentiate this process. The optimal conditions for the formation of volatile products of decay of chlororganic pesticides are created in dark chestnut and gray soils, derivatives of carbamine acids--in dark grey soil at humidity of 60-80% from the complete moisture capacity. Mineral fertilizers--ammoniacal nitrate and ammoniacal water, herbicides ronite and betanol--potentiate the decay of chlororganic pesticides, which can be used in the development of measures on soil purification from persistent pesticides.

Air Pollutants↗

[A pupillographical study on the presence of organochlorine pesticides in autonomic nerve disturbance].

It is well known that the effect of organophosphate pesticides on the autonomic nervous system is a cholinergic reaction. However no study has been done assessing the effect of organochlorine pesticides on the autonomic nervous system. We evaluated the autonomic nerve function using open-loop pupillography in 20 patients who had exposure to the organochlorine pesticides. Significant differences were seen in pupil area (p less than .006), velocity of both constriction and dilatation (p less than .001), and dilatation time (p less than .02), when compared with 18 normal subjects. Autonomic nerve disturbances were detected in eighteen of 20 patients (90%) by evaluating the pupillary light reflex in each patient. Sympathetic nerve inhibition i.e. sympatholytic pattern, was recognized in ten of 18 patients (55%). Four patients with sympatholytic pattern had a disturbance not only in the sympathetic nerve, but also parasympathetic nerve involvement suggesting pandysautonomia. The residue blood level of the organochlorine pesticides was examined in each patient. A high frequency of the DDE was recognized (85%). It was concluded that the toxicity of the organochlorine pesticide on the autonomic nerve appear as an inhibitory effect on pupil light reflex.

Adult↗

[Estimate of the dose of chlorinated pesticides consumed daily by the Italian population (1978-1984)].

This work reports an estimate of the doses of chlorinated pesticides taken with a total diet by the Italian population in the period 1978-84. Previous researches, carried out in 1971-72, had regarded both uncooked and cooked diets, ready for the consumption. It was found that the amount of the considered pesticides taken in total (HCB, gamma-BHC and total-BHC, Aldrin/Dieldrin, Heptachlor/Heptachlor-epoxide and DDT total-equivalent) calculated as a sum of the percentage of each acceptable daily intake (A.D.I.), at that time recommended was near the limit of acceptability (98.5% for cooked diets and 107.0% for uncooked diets). In addition it was found that nearly the 40% of the dose assumed in all came from milk and its derivates (butter, cheese). Then considering the contribution of milk and derivates to the amount of pollutants assumed with a total diet, a new evaluation has been done, with regard to the period 1978-84, i.e. after the prohibition of the use of these pesticides in Italy and in other industrialized countries. Then the data have been processed, with regard to 211 samples of foods (90 of milk, 45 of cheese, 76 butter) coming from determinations carried out by the Multizonal Laboratory of Hygiene and Prevention of Ferrara, Milano, Udine, Bolzano, Trento, Pescara and from the Institute of Hygiene of Rome University "La Sapienza" in the period mentioned. Furthermore, alimentary consumptions of milk, butter and cheese in 1978-84 have been evaluated, and amounts of chlorinated pesticides taken with these foods have been calculated. Finally the dose taken with these foods has been extrapolated to a total diet. It has come out that the doses (estimated) taken with the total diet have notably decreased between 1971-72 and 1978-84. An anomaly is yet represented by HCB: while the quantity taken with food has decreased from 1971-72 to 1978-84 (from 3.7 to 1.3 micrograms/man/day), the concentrations in human adipose tissues have increased (from 1.02 to 1.99 mg/Kg). It can reasonably mean that assumption of HCB is not prevalently alimentary, but that it is due to other factors of environmental contamination. The data of the doses of chlorinated pesticides taken have also been compared with the respective ADI indicated in 1971-72 and in 1978-84.

Animals↗

[Possibility of using immunologic indicators in evaluating the effect of pesticides on the body].

The modern problem of pesticide-related pathogenic body effect has been studied from different points of view. However, some medical aspects of pesticide-induced diseases, primarily their effect on the immune status, remain insufficiently explored. The study is designed to analyze pesticide effect on basic indicators of humoral and cellular immune factors and also to develop a new principle for assessing their exposure according to the degree of immune changes in those exposed to pesticides. Proceeding from the study results it is possible to conclude that under long-term exposure to pesticide microdoses the human body undergoes complex changes: there develops autoimmunization accompanied by the manifestation of antitissue autoantibodies, reduction of serum complement activity, relative content of T-lymphocytes and elevation of the level of B-lymphocytes. Toxicant-related body sensitization and decline in immuno-resistance can serve as the main cause for the deterioration of health status of the residents of the rural areas characterized by higher chemical exposure.

Adult↗

Environmental stress and parasitic infections: II. Effects of gramoxone and hexadrin (pesticides) on the survival characteristics of Fasciola gigantica miracidia.

Miracidia of Fasciola gigantica have a maximum longevity of 9 h. Miracidial survival was found to be age dependent and the instantaneous mortality rate increased exponentially with miracidial age and pesticide treatment. Mortality of the pesticide-treated miracidia differs significantly from that of the control. The LC50 value for Hexadrin was calculated at 1.93 ppm and for Gramoxone at 0.75 ppm. Dosage-mortality lines when tested statistically show no significant difference between the two pesticides at 95% level of confidence. The toxic effect of the pesticides was enhanced by synergism between the pesticides.

Animals↗

Effect of pH, light intensity and population on the toxicity of the pesticide carbofuran to the blue-green alga Nostoc muscorum.

The effect of pH, light intensity and population on the toxicity of the pesticide furadan (containing 3% carbofuran as the active ingredient) was studied on the nitrogen fixing blue-green alga Nostoc muscorum. It was observed that all these factors had effects on the toxicity of the pesticide. The blue-green alga grew better in the pH range of 7.5-10, whereas the acidic pH of 5-6 retarded growth. The pesticide was more toxic to alga in the medium of pH 5-6, whereas reduction in toxicity was noticed in alkaline pH of 7.5-10. The alga grew slowly in a light intensity of 1,500 lux in comparison to a light intensity of 3,000 lux, and no growth occurred in the dark. The toxicity of the pesticide gradually decreased with increasing light intensity. The toxic effect of the pesticide was increased when the initial population level was low, and increasing the initial population reduced the toxicity.

Carbofuran↗

Isolation and concentration of organophosphorus pesticides from drinking water at the ng/L level, using macroreticular resin.

A screening method has been developed for determining organophosphorus pesticides at ng/L levels in drinking water. Sixteen organophosphorus pesticides, diazinon, diazinon-oxon, dimethoate, ronnel, beta-phosphamidon, methyl parathion, ethyl parathion, malathion, chlorpyrifos, fenitrothion, ruelene, methidathion, ethion, EPN, phosalone, and phosmet, were extracted by Amberlite XAD-2 resin from 100 and 200 L drinking water previously spiked with these pesticides. The pesticides were eluted from the XAD-2 resin with acetone-hexane (15+85). The concentrated extract was analyzed by gas chromatography using a nitrogen-phosphorus selective detector and by gas chromatography-mass spectrometry using selected ion monitoring. Recoveries at the 10 and 100 ng/L spiking levels were greater than 90%, except recoveries for dimethoate and phosphamidon were 37 and 42%, respectively. The analysis of 300 L Ottawa tap water showed no detectable amounts (less than 1 ng/L) of any of the 16 organophosphorus pesticides.

Chromatography, Gas↗

[Adaptation to the action of several teratogens as a consequence of preliminary administration of pesticides to females].

The effect of DDT and lindane pesticides on the intensity of the teratogenic action of sodium acetylxalicylate (SA) and of the cabomate benlate group of pesticides was studied on Wistar rats given the mentioned pesticides from the onset of pregnancy. The teratogens were administered on the 10th and the 12th days of gestation, respectively. Preliminary administration of these pesticides was found to weaken the teratogenic and the embryotoxic action of benlate given in a dose of 250 mg/kg, and of SA administered in a dose of 400 mg/kg. When SA was given in a dose of 600 mg/kg preliminary administration of the pesticides decreased the postimplantation mortality of the embryos, but the number of fetuses with developmental anomalies was the same as in the isolated action of the preparation given in this dose.

Abnormalities, Drug-Induced↗

Organophosphate pesticide poisoning.

A total of 118 workers from a 120-person grape picking crew became ill in early September 1976. Of these (108 men and 10 women), 85 received medical attention and three of the 85 were admitted to hospital. The symptoms were typical for organophosphate poisoning. Average plasma and red cell cholinesterase values for the affected workers were depressed more than 60 percent. Most were treated with atropine and some were also treated with 2-PAM (pralidoxime). The exposure to residues of the organophosphate pesticides dialifor (Torak((R))) and phosalone (Zolone((R))) occurred in one grower's vineyards near Madera, California. It appeared that workers had been allowed into recently-treated areas before the expiration of the required 30-day safety interval for dialifor, and that excessive skin exposure to residues of this pesticide had resulted. The clinical management of these cases and the occupational surveillance of the workplace became quite complex. The grower sustained significant losses of grapes during the period in which some of his vineyards were under quarantine and he had to pay substantial medical expenses as well as a fine for violating state regulations concerning the proper use of pesticides. Organophosphate pesticides decay more slowly under hot, dry weather conditions than they do when rainfall is frequent. California has imposed a number of specific safety intervals to be observed after the application of these pesticides to certain crops. If, in violation of these regulations, workers are permitted to enter fields too soon, poisoning can occur.

Adult↗

Brain level and synthesis of acetylcholine during exposure to vibration and organic-phosphorus and phenol-derived pesticides.

The experiments were carried out on white rats studying the quantity and synthesis of acetyl-choline in the cerebral cortex and brain stem during exposure to vibration (50 Hz, 2 mm) and noise (80 dB), noise alone, and pesticides, chlorfenwinfos (1.66 mg/kg), DNPP (5.83 mg/kg), and DNBP (6.66 mg/Kg). The pesticides were given separately to resting animals and to animals exposed to vibration and noise, as well as noise alone. Vibration reduced the content and raised the synthesis of acetylcholine only in the cerebral cortex. The effect of exposure to noise included reduced content and increased synthesis of acetylcholine in the brain stem. Chlorfenwinfos, DNPP and DNBP had a similar effect raising the level of acetylcholine in the cortex and brain stem and decreasing the ability of its synthesis. Simultaneous exposure of the rats to the action of pesticides, vibration and noise, or noise alone led to mutual abolishing of the effects produced by each to these factors on the content and ability of acetylcholine synthesis in brain tissue. In the discussion it is suggested that vibration and noise stimulate the cholinergic centres in the brain. A concept of a mechanical action of vibration on the brain tissue is discussed also. Besides that, it was tried to explain the mechanism of action of pesticides on the content of acetylcholine and its synthesis and the mutual abolition of the effects produced by pesticides and those of vibration and noise, and noise alone on the concentration and ability of acetylcholine synthesis in the brain.

2,4-Dinitrophenol↗

Accumulation of anionic pesticides by rabbit choroid plexus in vitro.

In this study, two questions were addressed: 1) were anionic pesticides accumulated by choroid plexus via the organic acid transport system and 2) did such accumulation impair transport of anionic neurotransmitter metabolites normally eliminated from brain via this system? Rabbit lateral choroid plexi were incubated in Krebs-Ringer bicarbonate buffer in vitro with the herbicide, 2,4-dichlorophenoxyacetic acid, or the polar DDT metabolite, 2,2-bis (p-chlorophenyl)acetic acid (DDA). Both compounds were extensively accumulated. At a pesticide concentration of 1 microM, tissue-to-medium ratios of 40 (DDA) to 50 (2,4-dichlorophenoxyacetic acid) were found after 60-min incubation at 37 degrees C. Uptake of both compounds was energy- and concentration-dependent and could be inhibited by other organic anions. 2,4-Dichlorophenoxyacetic acid and DDA each produced dose-dependent inhibition of the uptake of the other pesticide. DDA was the more potent inhibitor. Thus, uptake of both compounds appeared to be mediated by the organic acid transport system. Furthermore, transport of the serotonin metabolite, 5-hydroxy-3-indole-acetic acid was more effectively inhibited by these pesticides than by the classical inhibitor, probenecid. Neither the parent amine, serotonin, nor the uncharged pesticides. DDT and dieldrin, inhibited 5-hydroxy-3-indole-acetic acid uptake. Thus, the effect appears specific for those compounds which share the organic anion transport system.

2,4-Dichlorophenoxyacetic Acid↗

Multicolumn solid-phase extraction cleanup of organophosphorus and organochlorine pesticide residues in vegetable oils and butterfat.

Diatomaceous earth columns used with reversed and normal solid phase extraction (SPE) cartridges were evaluated for the quantitative determination of a number of organophosphorus (OP) and organochlorine (OC) pesticide residues in edible vegetable oils and butterfat. An oil or fat sample (about 2 g) in hexane was passed through a diatomaceous earth (Extrelut QE) column and a C18 bonded silica (ODS) SPE cartridge, resulting in the separation of the pesticides from about 98% of the lipids. The eluate was split in half, with the first portion concentrated into acetone for the determination of OP pesticides by gas chromatography with flame photometric detection (GC-FPD). The other half was passed through an Alumina-N SPE cartridge for additional cleanup of lipid matrix to determine OC pesticides by GC with electron-capture detection (GC-ECD). Average recoveries from fortified samples were greater than 89% for the pesticides studied.

Butter↗

Concentrations of organochlorine pesticides in the adipose tissue of British Columbia residents.

Adipose tissue from a sample of 41 British Columbians was analyzed for a number of organochlorine pesticides. Pesticide concentrations were very low. Persistent fat-soluble compounds like DDT were found at the highest concentrations, whereas compounds that either quickly metabolize to other forms or are readily excreted were found at low levels or not detected. Data on age, sex, area of residence, occupation, diet and weight were obtained by interview. Increasing age was related to increasing levels of several of the persistent pesticides. Although there were insufficient data for statistical analysis, two individuals with potential occupational exposures had much higher levels of the pesticides than other subjects. Although population-based surveys of these organochlorine pesticides in human tissue may have little value, further study may be warranted for groups known to have had high exposures (e.g. occupationally exposed).

Adipose Tissue↗

Tank-mix adjuvants and pesticide residues: some regulatory and quantitative aspects.

Although it is well known that judicious use of adjuvants can increase the performance of foliage-applied sprays of many agrochemicals, little information is available in the public domain about their ultimate effects on pesticide residues in treated crops. In the present work, the influence of Agral (polyoxyethylene nonylphenols), Toil (methyl esters of rapeseed fatty acids) and Bond (styrene-butadiene copolymers) on surface and crop residues of diclofop-methyl/diclofop and propiconazole in wheat and field beans was investigated using a model system simulating field practice. Pesticides were applied as commercial formulations, diclofop-methyl 378 g litre(-1) EC (Hoegrass) and propiconazole 250 g litre(-1) EC (Tilt), at their maximum approved rates, 1135 g AI ha(-1) and 125 g AI ha(-1), respectively, both in the presence or absence of the maximum rate recommended for each candidate adjuvant. No detectable residues of diclofop-methyl or propiconazole were found in wheat 35 days after any of the four applications. However, residues of diclofop were present in this crop, and those from applications containing Agral (0.07 mg kg(-1) fresh weight (FW)) or Bond (0.08 mg kg(-1) FW) were significantly lower than those with no adjuvant (0.14 mg kg(-1) FW) or Toil (0.16 mg kg(-1) FW). Unlike wheat, residues of both diclofop and propiconazole were detected in field beans after harvest. Significantly higher residues of the former were recorded from the applications with Agral or Bond (ca 0.32 mg kg(-1) FW) than with those with no adjuvant or Toil (ca 0.15mg kg(-1) FW). All the propiconazole applications containing adjuvants showed a similar significant increase in residues (0.10-0.16 mg AI kg(-1) FW) over the no-adjuvant treatment (0.05 mg kg(-1) FW) in this crop. There appeared to be little agreement between the apparent amounts of uptake, as indicated by the rates of decline of surface residues up to 5 days after application, and final residues in either target species. On wheat, surface residues of diclofop-methyl decreased from initially ca 20 to as little as 0.02 mg kg(-1) FW using adjuvants; the corresponding values for propiconazole were ca 2 to ca 0.03 mg kg(-1) FW. Recoveries of diclofop-methyl from the surfaces of field beans were much higher than those from wheat, declining from ca 30 to only ca 6 mg kg(-1) FW during the course of 5 days; the corresponding residues for propiconazole were ca 2 mg to 0.15 mg kg(-1) FW. These findings are discussed in relation to uptake results obtained with radiolabelled pesticides and adjuvants in the laboratory, and to the mandatory requirements for pesticide residue data for the authorised use of adjuvants in the UK.

Agriculture↗

Ranking the risk of pesticide dietary intake.

Pesticide residues in various foods were assessed for the relative risk to the consumer. The analysis took into account criteria such as a pesticide's toxicity, usage, frequency of occurrence in foods, frequency of Maximum Residue Level (MRL) exceedances, and the overall risks of specific pesticide/food combinations. Examination of the top ten ranked pesticides for each criterion showed that there are no trends of commonality. Thus, no single pesticide is of particular concern from a consumer's point of view. This suggests that the consumer's risk perception is likely to be higher than justified.

Crops, Agricultural↗