Pesticides selectivity list to beneficial arthropods in four field vegetable crops.

Selectivity of pesticides to beneficial arthropods is a key data for the implementation of IPM program. In the context of field vegetables crop, a set of 16 fungicides, 17 herbicides and 14 insecticides commonly used in Belgium were tested on 5 indicator species: the parasitic hymenoptera Aphidius rhopalosiphi (De Stefani-Perez) (Hym., Aphidiidae), the aphid foliage dwelling predators Adalia bipunctata (L.) (Col., Coccinellidae) and Episyrphus balteatus (Dipt., Syrphidae) and the ground-dwelling predators Aleochara bilineata (Col., Staphyllinidae) and Bembidion lampros (Col., Carabidae). Pesticides were tested according a testing scheme including a first assessment on inert substrate (glass plates for adults of A. rhopalosiphi, larvae of A. bipunctata and E. balteatus, sand on adults of A. bilineata and B. lampros) and, for product that were toxic, a second assessment on natural substrate (barley seedlings for A. rhopalosiphi, french bean plants for A. bipunctato and E. balteatus and two type of soil for 8. lampros and A. bilineato). The effects of the product were assessed on basis on mortality, except for A. bilineata (Onion fly pupae parasitism). According to the final results obtained at the end of this testing scheme, the product were listed in toxicity class: green list if effect < or =30%, yellow list 30% < effect < 60% and orange list 60% < effect < or =80%. Products with toxicity higher than 80% on plants or on soils, or that reduce parasitism more than 80% on soil were put in red list and are not recommended for IPM. Results showed that all fungicides and herbicides were included in the green list except tebuconazole and boscalid + pyraclostrobin that were labeled as yellow for A. bipunctata. In opposite, no foliar insecticide was totally selective for all beneficial tested. However some products are in green list for one or several species. Soil insecticides were all are very toxic for ground dwelling arthropods and classed in red list. All results obtained during this study and further upgrade will be available on www.cra.wallonie.be/selectivite. In conclusions, fungicides and herbicides tested are compatible with IPM programs. For foliar insecticides, some treatments can be used carefully according to the selectivity. But for soil insecticide treatments, their toxicity raise the question of their use in IPM programs in vegetables and the need of new compounds or development of alternative pest control programs.

Influence of organic matter on bio-availability of carbosulfan and its toxicity on a carabid beetle.

Study of factors influencing soil insecticide toxicity are needed to reduce negative impacts of these products on beneficial insects. To date, if high toxicity differences between different type of soils have been reported, there is few specific studies on soil parameters influence on selectivity of soil insecticides to beneficial arthropods. To assess the specific impact of organic matter, the relationship between bio-availability of a soil insecticide, carbosulfan [Sheriff 1 Gr], and its toxicity on a small Carabidae, Bembidion lampros (Herbst.) on a sand enriched with increasing quantities of organic matter was studied. In laboratory, adults of B. lampros were put on different substrate, made of sand or sand with addition of organic matter at 3, 6 and 9% w/w, and treated with carbosulfan applied as granule at the rate of 312.5, 625, 1250 and 6250 microg a.i./m corresponding respectively to 0.5, 1, 2 and 10% of the recommended field rate. Mortalities of B. lampros were assessed after 14 day of exposure. In parallel, the total carbosulfan residue (total extraction) and bioavailable fraction (CaCL2 aqueous extraction) were determined 48h after substrate treatments. According to the mortalities and bio-availability obtained, a dose--response relationship was calculated and compared with a reference relation dose--response obtained on sand, where the bio-availability of the product was considered as 100% of the amount of product applied. Carbosulfan was highly toxic on sand for B. lampros, with 100, 57 and 50% mortality at 10, 2 and 1% of the recommended field rate. When organic matter was added to the sand, the toxicity gradually decreased. This reduction in toxicity was more rapidly observed on sand + organic matter than on pure sand. The mortalities were strongly correlated with the bioavailability, indicating first that the organic matter is fixing an important part of the insecticide and secondly reduce its toxicity to beneficial arthropods. The results suggest that it could be possible, with further research, to predict toxicity of products in the field on different kind of soil according previous laboratory toxicity models and soil analysis. In conclusion, the organic matter influences strongly bio-availability of carbosulfan. This bioavailability was strongly correlated to toxicity to B. lampros. With the improvement of bioavailability determination and method validation, the assessment of pesticide bio-availability in the substrate could help to estimate the pesticide toxicity towards carabidae on different type of soils.

New model for performance prediction in fixed-bed reactors based on the approach of the unused bed zone.

In the present study a practical model useful for designers dealing with pilot scale reactors for pollutant control has been developed. Investigations were undertaken for a single-component lead study on New Zealand clinoptilolite at the temperature of 25+/-1 degrees C. Fifteen runs under different operating conditions such as particle characteristics (expressed as d*(p)/D), column geometry (as l*/D) and flow speed (as R*e) have been performed. The results were interpreted by a response surface method (RSM) from which an equation giving the unused bed zone (UBZ) was obtained. From those dimensionless parameters, a sequence of contour plots was drawn, making it easy for a designer to choose optimum design parameters while controlling the operation performance. The flow rate (R*e) range was established over the laminar flow conditions from 1 to 8, (l/D*) was extended from 7 to 20 and (d(p)/D*) varied from 3% to 10%. Optimization of performance of the reactor as UBZ could vary from 10% to 40% of the material operating capacity.

Selectivity lists of pesticides to beneficial arthropods for IPM programs in carrot--first results.

In order to improve IPM programs in carrot, 7 fungicides, 12 herbicides and 9 insecticides commonly used in Belgium were tested for their toxicity towards five beneficial arthropods representative of most important natural enemies encountered in carrot: parasitic wasps - Aphidius rhopalosiphi (De Stefani-Perez) (Hym., Aphidiidae), ladybirds - Adalia bipunctata (L.) (Col., Coccinellidae), hoverfly - Episyrphus balteatus (Dipt.. Syrphidae), rove beetle - Aleochara bilineata (Col., Staphylinidae) and carabid beetle - Bembidion lampros (Col., Carabidae). Initialy, all plant protection products were tested on inert substrate glass plates or sand according to the insect. Products with a corrected mortality (CM) or a parasitism reduction (PR) lower than 30% were kept for the constitution of positive list (green list). The other compounds were further tested on plant for A. rhopalosiphi, A. bipunctata, E. balteatus and soil for B. lampros and A. bilineata. With these extended laboratory tests results, products were listed in toxicity class: green category [CM or PR < or = 30%], yellow category [30% < CM or PR < or = 60%] and orange category [60% < CM or PR < or = 80%]. Products with toxicity higher than 80% on plants or that reduce parasitism more than 80% on soil were put in red category and are not recommended to Integrated Pest Management programs in carrot. Results showed that all fungicides tested were harmless to beneficials except Tebuconazole, which was slightly harmful for A. bipunctata. Herbicides were also harmless for soil beneficials, except Chlorpropham. This product was very toxic on sand towards A. bilineata and must be tested on soil. All soil insecticides tested were very toxic for ground beneficials and considered as non-selective. Their use in IPM is subject to questioning in view of negative impacts on beneficials. Among foliar insecticides, Dimethoate and Deltamethrin are not recommended for IPM because their high toxicity for all beneficials. The other foliar insecticides were more selective; any of them were harmless for all species tested.

Combined enzymatic hydrolysis and HPAEC method for simultaneous analysis of galacturonic acid and neutral sugars of pectin.

Pectic substances are heteropolysaccharides from plant cell walls, which mainly consist of a homogalacturonan backbone of predominantly alpha-(1-->4) linked galacturonic acid (GalA) residues. This chain is interrupted by ramified rhamnogalacturonan regions with a certain amount of neutral sugars (rhamnose, arabinose, galactose, glucose, xylose and mannose) present as side-chains. The GalA residues can be methyl-esterified at the carboxyl group. Usually uronic acids of pectin are determined as anhydrogalacturonic acid by spectrophotometry, using the metahydrodiphenyl method (Thibault, 1979), while the monosaccharides are determined by High-Performance Liquid Chromatography (HPLC) after a methanolysis (Quemener and Thibault, 1990) or by Gaz Liquid Chromatography (GLC) as alditol acetates according to Blakeney et al. (1983). In this paper, a combined enzymatic hydrolysis and HPLC method is validated for simultaneous analysis of galacturonic acid and neutral sugars without any derivatisation.

Comparative activity of pulsed or continuous estradiol exposure on gene expression and proliferation of normal and tumoral human breast cells.

Intranasal administration of hormone replacement therapy presents an original plasma kinetic profile with transient estrogen levels giving rise to the concept of pulsed therapy. To further understand the molecular effects of this new therapy, we have compared the effects of pulsed and continuous estradiol treatments on two critical aspects of estradiol action: gene expression and cell proliferation. Cells were stimulated with estradiol as 1-h pulsed or 24-h continuous treatments at concentrations such that the 24-h exposure (concentration x time) was identical in both conditions. In MCF7 cells, the transcriptional activity of estrogen receptors (ER) on a transiently transfected responsive estrogen response element-luciferase reporter construct was shown to be drastically (approximately 10-fold) and similarly stimulated after both treatments. Moreover, the increased mRNA expression of three representative estradiol-sensitive genes (pS2, cathepsin D, progesterone receptor), evaluated by Northern blot, was identical after 1-h pulse with 7 nM estradiol or continuous treatment with 0.29 nM estradiol with the same kinetic profile over 48 h. Proliferation was quantified by a histomorphometric method on primary cultures of human normal breast cells from reduction mammoplasties and using a fluorescence DNA assay in six human breast cancer cell lines which were ER positive or negative. After a 7-day treatment period, estradiol had no effect on the proliferation of the three ER negative cell lines (BT20, MDA MB231, SK BR3) but significantly stimulated the proliferation of the normal cells and of the three tumoral hormone-sensitive cell lines (MCF7, T47D, ZR 75-1); both hormone treatments producing the same increases in cell growth. In conclusion, we have shown that the genomic or proliferative effects of estradiol were identical with pulsed or continuous treatments, thus indicating that estrogenic effects are not strictly related to concentrations but rather to total hormone exposure.

Analysis of 5-vinyl-1,3-oxazolidine-2-thione in complex matrices at ppb level.

5-Vinyl-1,3-oxazolidine-2-thione (5-VOT) is a goitrogenic compound released by enzymatic degradation of progoitrin, the most important glucosinolate occurring in rapeseed meal. This paper describes an analytical method for determining the 5-VOT in complex matrices. The method proposed by Quinsac et al. [J. Assoc. Off. Anal. Chem., 75(3) (1992) 529] has been improved by modification of the extraction conditions and the purification steps. The extraction of 5-VOT is performed with hot acid buffer. The first purification step is achieved by solid-phase chromatography (C(18)). The second purification step of 5-VOT is carried out by complexation with phenyl mercury acetate in cyclohexane and, afterwards, by decomplexation using an aqueous solution of sodium thiosulfate. These reactions move 5-VOT from an aqueous to an organic medium, and then back again to the aqueous phase. This procedure ensures a high purification efficiency. The precise quantification of 5-VOT is completed in 12 min by reverse-phase liquid chromatography (C(18)), using an isocratic elution with an ultraviolet detector and with synthetic 4,4-dimethyl-1,3-oxazolidine-2-thione as internal standard. Using this modified method, 5-VOT can be determined in different matrices such as rapeseed meal, animal diets, muscle, several organs (thyroid, liver, kidney and lung) and biological fluids (plasma and milk). The quantification limit of 5-VOT in the sample is of 1 ppb (1 mug kg(-1)), the recovery rate of 5-VOT is about 90% and the repeatability is over 97%.

Monophosphate 32P-postlabeling assay of DNA adducts from 1,2:3,4-diepoxybutane, the most genotoxic metabolite of 1,3-butadiene: in vitro methodological studies and in vivo dosimetry.

Among the main DNA-reactive metabolites of 1,3-butadiene (BD), both 1,2:3,4-butadiene diepoxide (BDE) and 1,2-epoxy-3-butene (BME) have been reported in mice and rats exposed to BD, but blood and tissue levels of these metabolites are much higher in mice than in rats under similar exposure conditions. BDE, being more reactive and genotoxic than BME, is thought to be responsible for the greater susceptibility of mice to BD carcinogenicity. While BDE is a DNA-alkylating agent and some BDE adducts have been characterized, no sufficiently sensitive method has been reported for studying BDE-DNA binding in vivo. In the present investigation, a modified dinucleotide/monophosphate version of the 32P-postlabeling assay was applied to detect BDE-DNA adducts, which were prepared by reacting BDE with calf thymus DNA or deoxyribooligonucleotides [(AC)10, (AG)10, (CCT)7 and (GGT)7] in vitro or with skin DNA of mice in vivo upon topical treatment. Optimal resolution by 2-D PEI-cellulose TLC of the highly polar 5'-monophosphate adducts was achieved at +4 degrees C using 0.3 M LiCI (DI) and 0.4 M NaCl, 0.04 M H3BO3, pH 7.6 (D2). The profiles of the 32P-postlabeled adducts were similar for calf thymus and skin DNA, with 3 major spots being detected. Adducts obtained in in vitro and in vivo experiments were compared by re- and cochromatography in 4 or 5 different solvents, and these experiments provided evidence that corresponding BDE adducts, for the most part, were identical and represented adenine derivatives. Guanine adducts were not detected by this method although literature data indicate their formation. Quantitatively, the assay responded linearly to adduct concentration, as shown in an experiment where BDE-modified skin DNA was serially diluted up to 81-fold with control DNA. The limit of detection was approximately 1 adduct in 10(8) normal nucleotides. Further, in an in vivo dosimetry study, skin DNA from groups of 8 individual mice treated with different doses of BDE (1.9, 5.7, 17, 51 and 153 mumol/mouse) for 3 days exhibited a linear relationship (r > or = 0.992) between adduct levels and dose. The results suggest that the 32P-postlabeling assay described herein will have utility in mechanistic studies and biomonitoring of DNA adduct formation from BDE and possibly other polar epoxides.

32P-postlabeling of 1,3-butadiene and 4-vinyl-1-cyclohexene metabolite-DNA adducts: in vitro and in vivo applications.

Epoxides of 1,3-butadiene, i.e. 1,2-epoxy-3-butene and 1,2:3,4-diepoxybutane are DNA-reactive metabolites. No methods have been reported for detecting DNA adducts of these compounds in exposed animals or humans. The purpose of this study has been to develop a 32P-postlabeling assay for adducts of 1,3-butadiene and its dimer, 4-vinyl-1-cyclohexene. The assay was applied to skin DNA of mice exposed topically to diepoxides of these dienes. A dose-dependent increase in in vivo adduct formation was observed for both compounds. The newly developed assay will find applications in mechanistic studies on these and related compounds and in human biomonitoring.

In vitro and in vivo (32)P-postlabeling analysis of 4-vinyl-1-cyclohexene (butadiene dimer) diepoxide-DNA adducts.

4-Vinyl-1-cyclohexene diepoxide (VCD), and industrial chemical, and its parent compound, 4-vinyl-1-cyclohexene (VCH), are potential health hazards, a they destroy oocytes in follicles in rodents. VCD is also a skin carcinogen at the site of application in both female and male mice and rats and after gavage, induces ovarian tumors in mice and forestomach tumors in rats. A (32)P-postlabeling assay was developed for the detection and measurement of VCD-DNA adducts. VCD, a direct-acting carcinogen, was reacted with DNA in vitro, as well as through mouse skin painting for 3 days with different doses of VCD. (32)P-Labeled adducts were separated by polyethyleneimine (PEI)-cellulose TLC and detected by screen-enhanced autoradiography. Comparable adduct profiles were obtained in vitro and in vivo. At higher doses (36-225 micro mol/mouse), adduct levels in vivo showed a linear dose response, while there was no difference between 14 and 36 micro mol/mouse. The limit of detection was estimated to be 1-3 adducts in 10(8) DNA nucleotides. The results show that VCD exposure gives rise to (presumably pre-mutagenic) DNA adducts in vivo and that (32)P-postlabeling can be applied to biomonitoring of VCD exposure.

Tamoxifen: evidence by 32P-postlabeling and use of metabolic inhibitors for two distinct pathways leading to mouse hepatic DNA adduct formation and identification of 4-hydroxytamoxifen as a proximate metabolite.

Exposure to pentachlorophenol (PCP) strongly intensifies the formation of mouse hepatic DNA adducts elicited by oral administration of tamoxifen (TAM), as previously shown by 32P-postlabeling. To explain this effect, PCP was proposed to interfere with the detoxication by sulfate conjugation of an as yet unidentified hydroxylated proximate TAM metabolite. A comparison of the present and earlier results shows that the hepatic TAM adduct pattern in female ICR mice depended on the route of administration of TAM (120 mumol/kg), with oral administration primarily eliciting formation of more polar adducts (termed group I adducts), while after i.p. administration less polar adducts (group II) predominated over group I adducts by a factor of 17.5. All these adducts were also formed in female Sprague-Dawley rats after i.p. dosing with TAM, but total adduct levels were 3.5- to 5-fold higher than in mice. After four daily i.p. treatments, TAM adducts accumulated in mouse liver DNA in a non-linear fashion. Adduct levels were 30-50 times lower in mouse kidney and lung than in liver. The phenolic metabolite 4-hydroxy TAM (120 mumol/kg) exclusively led to formation of polar (group I) hepatic adducts, and this process was stimulated 8-fold by co-administration of PCP (75 mumol/kg). Co-administration of PCP with the parent compound led to an 11-fold enhancement of group I adduct formation; simultaneously, levels of group II adducts were suppressed 6-fold. Another inhibitor of sulfate conjugation, 2,6-dichloro-4-nitrophenol, unlike PCP, had no effect on group I adducts, but it reduced group II adduct formation 2.2-fold. The PCP metabolite 2,3,5,6-tetrachlorohydroquinone (75 mumol/kg) did not significantly affect any major TAM adduct, suggesting that PCP itself was the active compound. Similar to group II TAM adducts, the formation of hepatic safrole-DNA adducts was inhibited in female ICR mice by both sulfotransferase inhibitors, consistent with the proposal that metabolic alpha-hydroxylation of the ethyl group of TAM followed by sulfate conjugation represented a mechanism of TAM activation. On the other hand, the strong intensification of group I adducts by PCP and the lack of this effect by 2,6-dichloro-4-nitrophenol suggested that inhibition of sulfate conjugation may not have been the primary mechanism underlying the intensification of group I adducts formed from TAM or 4-hydroxy TAM. The results presented herein demonstrate conclusively that TAM was activated to DNA-reactive compounds along two distinct pathways which contrasted in their responses to metabolic inhibitors.

Strong intensification of mouse hepatic tamoxifen DNA adduct formation by pretreatment with the sulfotransferase inhibitor and ubiquitous environmental pollutant pentachlorophenol.

Although negative in assays for mutagenicity, the clinically important antiestrogen tamoxifen induces hepatic DNA adduct formation in mice, rats and hamsters, as indicated by 32P-postlabeling, and is a potent hepatocarcinogen in rats. Both phenolic and alcoholic metabolites of tamoxifen have been reported. As these metabolites are potential candidates for sulfate conjugation, we examined whether the sulfotransferase inhibitor pentachlorophenol, a ubiquitous environmental contaminant, modulates hepatic tamoxifen adduct formation in vivo. Female ICR mice were given tamoxifen (45 mg/kg) daily per os for up to 4 days, with and without i.p. pretreatment with pentachlorophenol (20 mg/kg) 1 h before dosing with tamoxifen. At days 1, 2 and 4, liver DNA was analyzed 5 h after tamoxifen administration by a modified monophosphate version of the 32P-postlabeling assay. At day 4, pentachlorophenol pretreatment led to a large increase (13- to 17-fold) of the levels of four tamoxifen adduct fractions, while two adducts appeared unaffected, resulting in an approximately 7-fold enhancement of overall adduct formation. Significant pentachlorophenol related increases were also observed at day 1 and day 2. The mechanism of this effect has not yet been determined, but may involve the inhibition of sulfation of a tamoxifen metabolite(s) involved in the detoxication of the drug to nonelectrophilic derivatives. It was also apparent that there are two pathways of metabolic activation of tamoxifen, one being sensitive and the other resistant to pentachlorophenol.

[Monoxycarbogenic reaction of chloral: application to the obtention of carboxyhemoglobin and to its determination in blood]. / La réaction monoxycarbogénique du chloral: application à l'obtention de la carboxyhémoglobine et à son dosage dans le sang.

Carbon monoxide CO, generated from chloral (10(-2) M) in a dilution of hemochrome (pH 13), turns the solution of hemochrome into a solution of carboxyhemoglobin. The spectrum of this solution is identical to the one obtained after bubbling with CO. The reaction is fast and makes it possible to determine the percentage of carboxyhemoglobinemia in whole blood.