Analysis of organophosphorus and pyrethroid pesticides in organic and conventional vegetables using QuEChERS combined with dispersive liquid-liquid microextraction based on the solidification of floating organic droplet.

A simple, sensitive and environmentally-friendly method for determining organophosphorus and pyrethroid pesticides in vegetables was developed to better evaluate the risk of consuming them. The pesticides in vegetables were extracted, purified and concentrated by using the QuEChERS (quick, easy, cheap, effective, rugged and safe method) combined DLLME-SFO (dispersive liquid-liquid microextraction based on solidification of floating organic droplet) techniques. The key parameters were optimized through orthogonal array experimental design and statistical analysis. The linearity of the calibration curves was satisfied in matrix-matched standard solution with R2 ≥ 0.99. The limits of detection and limits of quantification were 0.3-1.5 and 0.9-4.7 µg/kg, respectively. The average recoveries of pesticides were 61.6-119.4% with relative standard deviations <16.1%. Furthermore, the method was applied successfully to analyse the pesticides in 15 pairs of organic and conventional vegetables. These results reflect the efficiency, reliability and robustness of the developed method.

Cohort-friendly protocol for the determination of two urinary biomarkers of exposure to pyrethroids and neonicotinoids using gas chromatography-triple quadrupole mass spectrometry.

Neonicotinoids (NEOs) and synthetic pyrethroids (PYRs) are active ingredients of commercial pesticides and/or insecticides with extensive indoor and outdoor applications, worldwide. Improved exposure metrics are warranted for NEOs and PYRs, if we are to better understand their human health effects. A cohort-friendly protocol for determining non-specific biomarkers of exposure to NEOs and PYRs, e.g. 6-chloronicotinic acid (6-CN) and 3-phenoxybenzoic acid (3-PBA), respectively, in human urine voids was proposed. A series of optimization experiments were conducted to validate the bioanalytical protocol using gas chromatography coupled with triple quadrupole mass spectrometry (GC-QqQ-MS/MS) in MRM mode. The method reached low detection limits for both analytes (0.075 µg L-1 for 6-CN and 0.050 µg L-1 for 3-PBA) in a short preparation and analysis time. The method used small initial urine sample volume (2 mL), short extraction time (≈ 240 min for the batches of 32 samples) and instrumental analysis time (≈ 14 min) for both pesticide metabolites in a single run. This protocol could facilitate the assessment of population exposure metrics for these pesticides and their inclusion in health risk assessment. Graphical abstract.

Resistencia a insecticidas en cepas de Aedes aegypti (Diptera: Culicidae) de tres distritos de la Región Pacífico Central de Costa Rica / nsecticide resistance in Aedes aegypti (Diptera: Culicidae) strains from three districts of the Central Pacific Region of Costa Rica

Introducción: Aedes aegypti es vector de virus dengue, chikungunya y Zika en Costa Rica. Su alta incidencia y carencia de vacunas hacen del control vectorial, incluyendo el químico, la única alternativa para disminuir su transmisión. El uso reiterativo de insecticidas propicia su resistencia. Objetivo: Determinar la resistencia y mecanismos de detoxificación enzimática a temefós e insecticidas piretroides en cepas de Ae. aegypti de tres distritos de la Región Pacífico Central de Costa Rica. Métodos: La resistencia a temefós, deltametrina y cipermetrina fue determinada en tres cepas de Barranca, Jacó y Quepos mediante bioensayos en larvas. Para cada insecticida se calculó la concentración letal 50 por ciento (CL50) y un factor de resistencia 50 por ciento(FR50), empleando la cepa Rockefeller como control. Ante la ocurrencia de resistencia, se repitieron los bioensayos utilizando butóxido de piperonilo, S,S,S, tributilfosforotritioato y ácido etacrínico que inhiben monooxigenasas, esterasas y glutatión S transferasa, respectivamente. Resultados: Ninguna cepa mostró resistencia a temefós. Las cepas Barranca y Jacó fueron resistentes a deltametrina (FR50= 7,38; 28,23, respectivamente). La cepa Jacó mostró, adicionalmente, resistencia a cipermetrina (FR50= 7,70). La cepa Quepos no mostró resistencia a ningún piretroide. Solo la cepa Barranca mostró disminución de resistencia a deltametrina asociada al butóxido de piperonilo denotando vinculación con monooxigenasas (FR50: 10,10). Para los otros casos de resistencia, ninguno de los sinergistas disminuyó la resistencia. Conclusiones: Las larvas de Ae. aegypti de las localidades evaluadas no mostraron resistencia a temefós. Respecto a piretroides se evidenció la aparición de resistencia que posiblemente no es mediada por detoxificación enzimática(AU)

Introduction: Aedes aegypti is the vector of dengue, chikungunya, and Zika viruses in Costa Rica. The high incidence and the lack of vaccines make vector control, including chemical control, the only measure to prevent transmission. The repetitive use of insecticides may induce resistance. Objective: To determine resistance and enzymatic detoxifying mechanisms to temephos and pyrethroids insecticides in strains of Ae. aegypti from three districts of the Central Pacific Region of Costa Rica. Methods: Resistance to temephos, deltamethrin, and cypermethrin was determined in three strains of Barranca, Jacó, and Quepos by larval bioassays. In each test, the lethal concentration 50 percent (LC50) and a factor of resistance 50 percent (FR50) were calculated, using the Rockefeller strain as control. When resistance was observed, the bioassays were repeated using piperonyl butoxide, S,S,S, tributylphosphorotritioate, and ethacrynic acid, that inhibit monoxygenases, esterases, and glutathione S- transfererase, respectively. Results: None of the strains were resistant to temephos. Resistance to deltamethrin (FR50= 7.38 and FR50= 28.23, respectively) was determined in the strains from Barranca and Jacó, while resistance to cypermethrin was detected only in Jacó (FR50= 7.70). The Quepos strain was not resistant to any pyrethroid. Only the Barranca strain showed a decrease in the resistance to deltamethrin when piperonyl butoxide was used, linking the resistance to monooxygenase enzymes (FR50: 10.10). For the other cases, none of the synergists decreased the resistance. Conclusions: Larvae of Ae. aegypti from the localities evaluated were not resistant to temephos. With respect to pyrethroids, results show an emergence of resistance that may not be mediated by enzymatic detoxification(AU)

Cyphenothrin Flea and Tick Squeeze-On for Dogs: Evaluation of Potential Health Risks Based on the Results of Observational Biological Monitoring.

An observational biomonitoring study was conducted involving adults and children in households that purchased and applied a cyphenothrin-containing spot-on product for dogs as part of their normal pet care practices. The 3- to 6-yr-old children had greater exposure than the adult applicators in the same house, 3.8 and 0.6 µg/kg body weight, respectively. The mean measured values in children were 13-fold lower than those estimated using the U.S. Environmental Protection Agency (EPA) current standard operating procedures (SOP) for pet products (assuming 5% dermal absorption), although the maximum absorbed dosage of one child on one day was equivalent to the default value derived from the SOPs. With regard to potential human health risks, it can be concluded that despite the inherent conservatism in both the exposure and toxicology data, the margins of exposure (MOE) were consistently greater than 100 for average, 95th percentile, and maximum exposures. More specifically, the results of this study demonstrated that the MOE were consistently greater than 1,000 for mean exposures and exceeded 100 for 95th percentile and maximum measured exposures, which clearly indicates a reasonable certainty of no harm when using the cyphenothrin spot-on products. It is also noteworthy that Sergeant's spot-on products containing cyphenothrin currently sold in the United States have lower weight percentages of active ingredient and lower applied amounts than those used by all but two of the participant households in this study.

Estimation of measurement uncertainty associated to the determination of pesticide residues: a case study.

In the analytical analysis the measurement uncertainty is a quantitative indicator of the confidence describing the range around a reported or experimental result within which the true value can be expected. Several approaches can be used to estimate the measurement uncertainty associated to the analysis of pesticide residues: a) the top-down, the estimation can be referred to default values; b) the bottom-up the estimation is related to the uncertainty sources. Concerning the bottom-up approach, the following contributions have been investigated: weight of sample, calibration solutions, final volume of sample and intermediate repeatability studies. The commodity/residue combination selected in this study was celery/tau-fluvalinate pesticide. Tau-fluvalinate is a broad-spectrum insecticide in the pyrethroid class of pesticides. The Maximum Residue Limit (MRL) of tau-fluvalinate in celery has been set at 0.01 mg/kg. The tau- Fluvalinate showed two chromatographic peaks. Since the individual standards are not available, the two peaks were integrated separately and the instrumental responses were added. The total residue was calculated on the basis of resulted peaks. The present work aims to compare the uncertainty estimated by experimental data using repeated analysis (n = 12) of a real sample and a spiked sample. The relative expanded uncertainty for two data set, incurred and spiked, was 22 % and 20 %, respectively. No differences were observed from repeated determinations of real samples and spiked samples.

Application of the industrial hygiene hierarchy of controls to prioritize and promote safer methods of pest control: a case study.

In 2005, the California Department of Public Health, Occupational Health Branch (OHB) investigated an incident of pesticide exposure and identified 27 vineyard workers who became ill due to drift of cyfluthrin, a pesticide being applied to a neighboring orange field to control katydids. Another pest, citrus thrips, was also present in the field. We investigated safer alternatives for katydid and thrips control to prevent illness due to pesticide exposure and used the industrial hygiene hierarchy of controls to prioritize the control methods. OHB evaluated factors that contributed to pesticide exposure and identified safer alternatives by conducting literature reviews on katydid and thrips control, drift prevention technology, and other relevant topics, and by interviewing integrated pest management advisors, conventional and organic growers, equipment manufacturers, county agricultural commissioners, pest control advisors, regulatory agencies, and others. We prioritized methods using the industrial hygiene hierarchy of controls. We identified safer pest control practices that incorporated hazard elimination, chemical substitution, engineering controls, and administrative controls, including employer policies and government regulations.

Dyeing process may alter the efficacy of insecticide-treated nets.

The biological efficacy as judged by mortality against Anopheles gambiae s.s. of three net samples, one white and two colored, treated with deltamethrin by conventional dipping was compared. Efficacy as well as chemical analysis results showed that uptake of insecticide by white net material was much higher than by colored nets. After a single wash, efficacy of colored nets was reduced significantly below 80% mortality, which is the minimum acceptable level for a field application. This unexpected result could be because of either low uptake of insecticide by colored fibers or high alkalinity on the fibers, resulting from the dyeing process (reduction clearing). alpha-Cyanopyrethroids, such as deltamethrin, have a low stability in alkaline conditions and rapidly degrade. Practical implications of this finding are of primary importance because the majority of nets currently purchased by institutional buyers for malaria prevention are colored.

Determination of eight synthetic pyrethroids in bovine fat by gas chromatography with electron capture detection.

Synthetic pyrethroids are among the most widely used classes of insecticides, and their uses are varied, including plant protection, animal dips, and as a treatment for human clothing and bedding in very hot climates. Veterinary applications include ear tags, pour-on formulations, sprays, and dips. Persistent residues have been reported in livestock, and routine monitoring programs in other countries have found detectable residues of various pyrethroids in fat. A method has been developed using solid-phase extraction that reduces the quantities of solvents used, the time required, and the amount of glassware used compared to an earlier method on which it was based. The scope of analytes tested included the 5 compounds cited in the earlier method (flucythrinate, permethrin, cypermethrin, fenvalerate, and deltamethrin) and, in addition, cyfluthrin, lambda-cyhalothrin, and fluvalinate. Sample extracts were analyzed by gas chromatography with electron capture detection using selected chromatographic peaks characteristic of each compound. Limits of quantification for the compounds were from 25-50 microg/kg, with a linear response for all compounds to 200 microg/kg. Recoveries ranged from 80 to 123%.

Assessment of insecticide contamination in runoff and stream water of small agricultural streams in the main soybean area of Argentina.

The first- and second-order streams, Brown and Horqueta, respectively, which are located in the main area of soybean production in Argentina were examined for insecticide contamination caused by runoff from nearby soybean fields. The insecticides most widely used in Argentina (chlorpyrifos, cypermethrin and endosulfan) were detected in sediments, suspended particles and water. Highest concentrations in suspended particles were 318 microg/kg for endosulfan in the stream Horqueta, while 226 microg/kg chlorpyrifos and 13.2 microg/kg cypermethrin were measured in the stream Brown. In the Horqueta stream 150 and 53 microg/kg chlorpyrifos and cypermethrin were detected in runoff sediments, respectively. Whereas cypermethrin concentrations in the suspended particles were relatively low, levels in the floodwater of Brown reached 0.7 microg/l. The highest chlorpyrifos concentration in floodwater was 0.45 microg/l in Brown. However, endosulfan was not detected in the water phase. In runoff water the highest concentrations measured were 0.3 microg/l for chlorpyrifos in Horqueta and 0.49 microg/l for cypermethrin in the Brown stream. On five sampling dates during the pesticide application period in Brown stream (2002/2003) the concentration of chlorpyrifos and cypermethrin in runoff and/or floodwater exceeded the water quality criteria for freshwater mentioned in this study. In three cases this insecticide concentration was measured in stream water, indicating an acute risk to aquatic life. The acute toxicity-exposure-ratio (TER) for chlorpyrifos and cypermethrin also shows an acute risk for aquatic invertebrates in the Brown stream. In the Horqueta chlorpyrifos concentrations in the runoff exceeded the safety levels three times during the application period (2001/2002), potentially endangering the aquatic fauna. Effects on aquatic macroinvertebrates after insecticide contamination were reported in earlier studies in Horqueta stream.

Evaluation of safety and efficacy of acaricides for control of the African tortoise tick (Amblyomma marmoreum) on leopard tortoises (Geochelone pardalis).

Nine acaricides (amitraz, carbaryl, chlorpyrifos, cyfluthrin, fipronil, lindane, permethrin, phenothrin, and pyrethrins) were studied for their efficacy in killing the African tortoise tick (Amblyomma marmoreum). Only four of the acaricides (chlorpyrifos, cyfluthrin, lindane, and permethrin) produced 100% mortality within 24 hr of application, and only two (cyfluthrin and permethrin) continued to cause 100% mortality when diluted to as low as 1:10,000. Five of the acaricides (amitraz, carbaryl, chlorpyrifos, cyfluthrin, and permethrin) were studied for toxicity to the leopard tortoise (Geochelone pardalis), the most common host of A. marmoreum. The results indicate that cyfluthrin and permethrin, which were herein found to be the two most effective acaricides for control of A. marmoreum, are also the safest acaricides for use on leopard tortoises.

[An experimental tool essential for the evaluation of insecticides: the testing huts]. / Un outil expérimental indispensable à l'évaluation des insecticides: les cases-pièges.

The following study analyses the potentialities of the experimental huts built in M'be Valley (Côte d'Ivoire) where the evaluations of the insecticide products have been carried out for many years in line with the WHOPES protocol on the methodology of stage 2 assays. Starting a testing station first requires a good knowledge of the sensitivity of Anopheles gambiae to the main insecticide families. Then thanks to the experimental huts the efficacy of the various means of treatment can be compared with the one in untreated huts; this study focuses on house spraying using 100 mg a.i./m2 and bednets impregnated with lambda-cyhalothrin at a dose of 15 mg a.i./m2. The fipronil used in house spraying doesn't show any repellent effect, however it does have an irritating effect that increases the natural exophily of An. gambiae females entering the testing huts. The blood-feeding rate recorded in the treated huts was reduced to 24% and to 38% mortality rate consisting mainly of a 24 hours delayed mortality. The bednets treated with lambda-cyhalothrin have greatly reduced the contact between man and vector since the entry rate of An. gambiae females was cut down by 68% compared to the control. The exophily of this anopheles was twofold greater with the impregnated bednets and the blood-feeding rate reduced to 47%. Finally the global mortality rate, two thirds of immediate mortality, one third of delayed mortality, reached 35%. The experimental huts in the M'be Valley therefore provide essential information regarding the selection of the most efficacious insecticides against An. gambiae. This experimental method must be extended to other sites in order to finalize ever more selective and appropriate means of control against nuisance and disease-vector mosquitoes.

Chemistry and specifications of pesticides.

This report presents the conclusions of a WHO Expert Committee commissioned to make recommendations on specifications for pesticides used in public health, with a view to promoting the manufacture and use of high-quality products that are both effective against susceptible vectors of disease and acceptable in terms of safety. These specifications serve mainly for purchase and quality-control purposes, but may also be useful for product registration by WHO Member States. The text includes an outline of the objectives and achievements of the WHO Pesticide Evaluation Scheme (WHOPES) and an overview of recent trends in pesticide usage in the various WHO regions. Analytical methods and quality control of pesticides in developing countries are discussed, and the Committee's recommendations for specifications for various types of pesticides are presented. The Committee also considered pesticide containers and packaging, marking and storage, as well as the problems of disposal of unusable pesticides and their containers. Recommendations for continued collaboration between WHO and other organizations are summarized, as are the Committee's more general recommendations for WHO's future work in the area of pesticides. Annexed to the report are the Committee's recommendations for changes to existing specifications and methods, for new pesticides and formulations, and for new methods.

The efficacy and residual life span of two alphacypermethrin insecticide formulations (Fendona 6% suspension concentrate and Fendona Dry 15%) treated on mosquito bed nets.

OBJECTIVE: To compare the effects of mosquito nets treated with two formulations of alpha-cypermethrin insecticide with a view to recommending the most appropriate formulation for use to treat mosquito nets. DESIGN: Assessment of insecticide potency under conditions of ordinary use. SETTING: Chilonga rural irrigation community in south east Lowveld Zimbabwe, a holoendemic area of year round malaria transmission. SUBJECTS: Convenience sample of 20 household heads (volunteers) was issued with treated mosquito nets. INTERVENTION: Following mosquito net treatment and distribution, bioassay tests were carried out monthly for a period of six months on insecticide and placebo treated nets. Questionnaires were administered once, one month post treatment of nets. MAIN OUTCOME MEASURES: Median mosquito knock down times of mosquitoes exposed to insecticide treated nets and community attitude towards the use of insecticide treated nets. RESULTS: Average time taken to knock down the median mosquito progressed from 2.3 minutes to 13.0 minutes for Fendona Dry 15% and from 4.1 minutes to 7.8 minutes for Fendona 6% SC over the six month period. The average time taken to knock down the median mosquito three months post-washing were 13.0 minutes and 7.4 minutes for Fendona Dry 15% and Fendona 6% SC respectively as against more than 30 minutes in controls. Both insecticides exhibited some wash resistance properties. The side effects reported were sneezing, itching, skin rash and smelling. Questionnaire data suggested that alphacypermethrin treated mosquito nets were welcomed. CONCLUSIONS: Both insecticide formulations proved to be suitable candidates. This was because of their insecticidal potency, wash resistance properties and acceptance by the community.

Cypermethrin pour-on synergized with piperonyl butoxide: effects on Haematobia irritans (Diptera: Muscidae) natural populations resistant to cypermethrin.

Development of pyrethroid resistance in Haematobia irritans in Santa Fe province, Argentina, resulted in an increased use of pyrethroid insecticides, probably due to lack of suitable alternative treatments. We explored the efficacy of mixtures of cypermethrin and piperonyl butoxide (PBO) against pyrethroid-resistant H. irritans. Groups of 25 Holstein cows each, naturally infested with cypermethrin resistant H. irritans were assigned to treated or control groups in April, September, October and December 1997. Cattle in treated groups were medicated with pour-on oil formulations of 5% cypermethrin (dose = 4 mg per kg of body weight) with 5% or 10% PBO in April, and with a mixture containing 5% of both components thereafter. Efficacy was tested for 21 days after treatment. A treatment of 5% cypermethrin pour-on without PBO was evaluated in October 1997. Samples of horn flies were obtained before September, October and December treatments and exposed for 2 h to filter papers impregnated with different cypermethrin concentrations to determine the 50% lethal concentration (LC50). No difference in efficacy was found between cypermethrin pour-on formulations with 5% or 10% of PBO (more than 94% efficacy on day 21 after treatment). Efficacy of 5 % cypermethrin-5% PBO mixture decreased rapidly in the successive treatments (less than 40% efficacy was observed on day 21 after the December treatment), and the period after treatment with an efficacy higher than 95% was 14 days for the treatment carried out in April, 10 days in September; 7 days for the treatment performed in October and 4 days for the December treatment. The LC50 of cypermethrin was 36.6 microg per cm2 in September and increased to 116.6 and 226.1 microg per cm2 in October and December, respectively. It is concluded that the addition of PBO to cypermethrin did not provide a treatment that would give a long term control of pyrethroid resistant-horn flies.

Biological activity of permethrin, phenothrin/allerthrin and D-phenothrin on Periplaneta americana and Blattella germanica cockroaches.

The biological activity of permethrin, phenothrin/allethrin and d-phenothrin was evaluated against Periplaneta americana and Blattella germanica cockroaches. A decrease in the number of cockroaches caught after spraying the trial huts was observed. The decrease may be attributed to either the presence of trap-weary specimens or because P. americana is not fully attracted by left over food. One hundred percent knock down was achieved after 10 minutes for all insecticides. The knock down rates for d-phenothrin and permethrin were similar as compared to a combination of phenothrin/allethrin. However, all insecticides were not very effective against cockroaches because the field residual effect was from one day to two days.

Settling of insecticide from dip wash mixed with dam water and zinc sulphate and used to control sheep lice (Bovicola ovis).

Insecticidal dipping fluid emulsions, mixed in vitro in dam water containing suspended clay particles and 1% w/v zinc sulphate, were analysed to determine rates of settling of diazinon, cyhalothrin and cypermethrin. Fifteen minutes after mixing, the concentration of the insecticides 5 cm below the surface had declined by 72.5%, 72.8% and 89.4%, respectively. On remixing, the concentration of insecticide in suspension was close to or greater than the initial concentration. In 2 trials, lice were eradicated from sheep showered with dip wash mixed in cloudy dam water to which 1% w/v of zinc sulphate was added. In 12 flock treatments in which 1000 to 2000 sheep were dipped with added zinc sulphate, the concentration of insecticide remained above the minimum lethal for susceptible strains of lice. However, lice were still present 6 months later in 8 of these flocks. When zinc sulphate is added to dip wash, agitation is needed to maintain the insecticide in suspension.

[Toxicology and hygienic standardization of the new insecticide neopinamine]. / Toksikologiia i gigienicheskoe normirovanie novogo insektitsida neopinamina.

Proceeding from the experimental study of neopinamine insecticide toxicity its hygienic standardization was carried out. Since neopinamine had moderate toxicity under its acute and chronic effect and different routes of body intake, its accumulation was absent and its impact was selective in the occurrence of aftereffects, this preparation was attributed to the third class of danger. Neopinamine MAC in the workplace air was recommended at the level of 5 mg/m3.