Resistance to pirimiphos-methyl in West African Anopheles is spreading via duplication and introgression of the Ace1 locus.

Vector population control using insecticides is a key element of current strategies to prevent malaria transmission in Africa. The introduction of effective insecticides, such as the organophosphate pirimiphos-methyl, is essential to overcome the recurrent emergence of resistance driven by the highly diverse Anopheles genomes. Here, we use a population genomic approach to investigate the basis of pirimiphos-methyl resistance in the major malaria vectors Anopheles gambiae and A. coluzzii. A combination of copy number variation and a single non-synonymous substitution in the acetylcholinesterase gene, Ace1, provides the key resistance diagnostic in an A. coluzzii population from Côte d'Ivoire that we used for sequence-based association mapping, with replication in other West African populations. The Ace1 substitution and duplications occur on a unique resistance haplotype that evolved in A. gambiae and introgressed into A. coluzzii, and is now common in West Africa primarily due to selection imposed by other organophosphate or carbamate insecticides. Our findings highlight the predictive value of this complex resistance haplotype for phenotypic resistance and clarify its evolutionary history, providing tools to for molecular surveillance of the current and future effectiveness of pirimiphos-methyl based interventions.

Chronic pesticide mixture exposure including paraquat and respiratory outcomes among Colombian farmers.

This study explored the potential association between chronic exposure to pesticide mixtures including paraquat and respiratory outcomes among Colombian farmers. Sociodemographic and occupational data, respiratory symptoms and spirometric data were collected. Paraquat in spot urine samples were quantified with solid-phase extraction high-performance liquid chromatography. Multiple Poisson regressions with robust variance were used to determine factors associated with respiratory outcomes. Profiles of pesticide mixtures used were identified among 217 farmworkers, but profenofos and methamidophos-based mixtures were more frequent. Chronic paraquat exposure was slightly associated with self-reported asthma (PR: 1.06; 95% CI 1.00 to 1.13). Different pesticide mixtures were associated with flu, thoracic pain, allergic rhinitis, and obstructive pattern in spirometry. Although acute exposure to paraquat is low among Colombian farmers participating in the study, associations between respiratory outcomes and chronic pesticide mixtures exposure including profenofos, methamidophos or glyphosate require further specific studies.

Part 2: Forensic attribution profiling of Russian VX in food using liquid chromatography-mass spectrometry.

This work is part two of a three-part series in this issue of a Sweden-United States collaborative effort towards the understanding of the chemical attribution signatures of Russian VX (VR) in synthesized samples and complex food matrices. In this study, we describe the sourcing of VR present in food based on chemical analysis of attribution signatures by liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with multivariate data analysis. Analytical data was acquired from seven different foods spiked with VR batches that were synthesized via six different routes in two separate laboratories. The synthesis products were spiked at a lethal dose into seven food matrices: water, orange juice, apple purée, baby food, pea purée, liquid eggs and hot dog. After acetonitrile sample extraction, the samples were analyzed by LC-MS/MS operated in MRM mode. A multivariate statistical calibration model was built on the chemical attribution profiles from 118 VR spiked food samples. Using the model, an external test-set of the six synthesis routes employed for VR production was correctly identified with no observable major impact of the food matrices to the classification. The overall performance of the statistical models was found to be exceptional (94%) for the test set samples retrospectively classified to their synthesis routes.

Pesticide residues in leafy vegetables and human health risk assessment in North Central agricultural areas of Chile.

To investigate pesticide residue concentrations and potential human health risk, a study was conducted in 118 leafy vegetable samples collected in 2014-2015 from the North Central agricultural areas of Chile. The pesticide residues were determined using the multiresidue QuEChERS method by gas chromatography as well as high-performance liquid chromatography. The results indicated that 27% of the total samples contained pesticide residues above the maximum residue limits of each active ingredient. The maximum estimated daily intake obtained for carbon disulphide (CS2), methamidophos, azoxystrobin and cypermethrin were 0.57, 0.07, 0.06 and 0.05 mg kg-1, respectively, which was higher than their acceptable daily intake. It is concluded that inhabitants of the North Central agricultural area of Chile are not exposed to health risks through the consumption of leafy vegetables with the exception of methamidophos. Nevertheless, the high levels of methamidophos detected in leafy vegetables could be considered a potential chronic health risk.

Pharmacokinetics and effects on serum cholinesterase activities of organophosphorus pesticides acephate and chlorpyrifos in chimeric mice transplanted with human hepatocytes.

Organophosphorus pesticides acephate and chlorpyrifos in foods have potential to impact human health. The aim of the current study was to investigate the pharmacokinetics of acephate and chlorpyrifos orally administered at lowest-observed-adverse-effect-level doses in chimeric mice transplanted with human hepatocytes. Absorbed acephate and its metabolite methamidophos were detected in serum from wild type mice and chimeric mice orally administered 150mg/kg. Approximately 70% inhibition of cholinesterase was evident in plasma of chimeric mice with humanized liver (which have higher serum cholinesterase activities than wild type mice) 1day after oral administrations of acephate. Adjusted animal biomonitoring equivalents from chimeric mice studies were scaled to human biomonitoring equivalents using known species allometric scaling factors and in vitro metabolic clearance data with a simple physiologically based pharmacokinetic (PBPK) model. Estimated plasma concentrations of acephate and chlorpyrifos in humans were consistent with reported concentrations. Acephate cleared similarly in humans and chimeric mice but accidental/incidental overdose levels of chlorpyrifos cleared (dependent on liver metabolism) more slowly from plasma in humans than it did in mice. The data presented here illustrate how chimeric mice transplanted with human hepatocytes in combination with a simple PBPK model can assist evaluations of toxicological potential of organophosphorus pesticides.

Use of cholinesterase activity as a biomarker of pesticide exposure used on Costa Rican banana plantations in the native tropical fish Astyanax aeneus (Günther, 1860).

In Costa Rica, thousands of tones of agricultural pesticides have been used for decades and their use is continuously increasing due to intensive and expanding production of coffee, pineapple, rice, ornamental plants and bananas. The objective of this study was to evaluate whether choline esterase (ChE) activity could be used as a biomarker of exposure to pesticides in the Costa Rican native fish Astyanax aeneus (characidae). Three methods used in order to evaluate the ChE biomarker were as follows: Laboratory studies where A. aeneus was exposed to organophosphate pesticide (ethoprophos); In situ 48 hr exposure assessment using caging experiments with fish exposed upstream and downstream of banana plantations and ChE activity estimation of in fish captured directly at sites with different degrees of pesticide exposure. Results from the laboratory studies showed that ChE activity in both brain and muscle tissue was significantly lower in fish exposed to ethoprophos than in controls. Fish from the caging experiments showed no difference in ChE activity neither in brain nor in muscle tissue between the four tested sites and was attributed to the short duration of the exposure. Asignificant difference in ChE activity was determined in muscle of fish captured from Laguna Madre de Dios compared to fish from Canal Batán. Although our laboratory results revealed that ChE activity in A. aeneus was highly responsive to ethoprophos, results from field experiments were less conclusive and showed that the captured fish showed large variability in ChE activity and that more research is needed before ChE activity can be used as reliable biomarker of pesticide exposure.

Adhesion- and migration-related side effects of phosphothioated CpG oligodeoxynucleotides.

Nucleic acid oligodeoxynucleotides (ODN) are increasingly used in biological research and in clinics where they are used for both diagnostic and therapeutic purposes. In order to increase the stability and efficacy of ODNs, various chemical modifications have been applied to create nucleic acid derivatives that are not recognized by endogenous nucleic acid cleavage mechanisms. One of the most common and cost-effective modifications is the phosphothioate (PTO) modification. The PTO modification is implemented mainly in antisense ODN, but also in ODN that were shown to activate members of the toll-like receptor (TLR) family such as TLR3 (poly-I:C), TLR8 (ssRNA) and TLR9 (CpG). We recently found that PTO-ODN aimed at activating TLR9 induce a non-TLR9-specific detachment phenotype in a growth-substrate dependent manner. Moreover, we found that unmodified and PTO-modified TLR ligands induce distinct patterns of gene expression in cultured neural cells. These findings suggest that PTO-ODN can cause nonspecific effects on cell adhesion that could compromise interpretation of data from experiments using PTO-ODN.

Effects of pirimiphos-methyl (an organophosphate insecticide) on the fertility of adult male rats.

BACKGROUND: Organophosphate insecticides represent one of the most widely used classes of pesticides with high potential for human exposure in both rural and residential environments. OBJECTIVE: In the present study, we investigated the effects of pirimiphos-methyl (0, 2-diethylamino-6-methylpirimidin-4-yl O, O-dimethyl phosphorothioate), an organophosphothioate pesticide, on male rat reproductive performances. METHODS: A total of 24 adult Wistar rats were divided into 4 groups of 6 animals each and orally treated with 0, 41.67, 62.5 or 125 mg/kg of pirimiphos-methyl for 90 days. RESULTS: Results from the study showed a significant increase (p<0.05) in feed consumption, body weight gain, relative testis and epidiydimis weights and intra-testicular cholesterol level in rats receiving the test substance at doses of 62.5 or 125 mg/kg whereas a significant decrease (p<0.05) in serum total protein, sperm density and motility, fertility and parturition indices and pups sex-ratio (M/F) was recorded in animals treated with 125 mg/Kg of pirimiphos methyl. Histological findings also indicated enlargement of interstitial space, inhibition of spermatogenesis, rarefaction of Leydig cells and oedema in testes compared to control animals. CONCLUSION: It could then be concluded that pirimiphos-methyl (62.5 and 125 mg/kg) is detrimental to the reproductive potentials of male rats.

Effects of separate and combined exposure to the pesticides methamidophos and chlorothalonil on the development of suckling rats.

It has been suggested that, in some cases, exposure to environmental contaminants affects children more profoundly than adults. It is important to evaluate adverse health outcomes in children, a population susceptible to toxic chemicals and mixtures. We examined the effects that maternal exposure to two pesticides had on maturational aspects of offspring development during the nursing period. Nursing female rats were exposed to 1-4mg/kg of intraperitoneal methamidophos, 200-800mg/kg of chlorothalonil, or both. The higher doses of methamidophos affected pup viability by day 21 of life. Both pesticides, alone or together, affected body weight gain of dams and offspring. Developmental milestones evaluated in the pups were incisor eruption, ear unfolding, eye opening and testis descent. Although no clear dose-response relationship was established between these milestones and exposure to methamidophos or chlorothalonil, incisor eruption was accelerated in many groups, and the majority of rat offspring exposed to methamidophos presented later ear unfolding and eye opening than did the control group offspring. Sexual maturation (testis descent) was significantly delayed in some groups. For dams and pups alike, simultaneous exposure to both pesticides was not found to have a greater toxic effect than that resulting from exposure to only one of the two. Taken together, these results demonstrate exposure-related influences on several developmental measures. Detection of more subtle effects may be improved through the use of the developmental temporal response protocols utilized in this study.

[Effects of acetochlor and methamidophos on culturable fungal population and community structure in black soil].

By the methods of CFU and rDNA-PCR-DGGE, this paper studied the dynamic changes of culturable fungal populations, community structure, and population richness in black soil under stress of acetochlor and methamidophos. The results showed that the growth of fungi was promoted under lower concentration acetochlor (50 mg x kg(-1)), but long-standing inhibited under its higher concentration (250 mg x kg(-1)). 250 mg kg(-1) methamidophos promoted the growth of culturable fungi intensively during 8-week period, resulting in 10 times increase in CFU of fungi than the control, while low concentration methamidophos (50 mg x kg(-1)) had no evident effect. The richness of culturable fungi in soil treated by medium and high concentration acetochlor was decreased irreversibly. Cluster analysis of fungal rDNA-PCR-DGGE fingerprints profile showed that acetochlor and methamidophos could change the culturable fungal community structure diversely, and the effect of methomidophos was more obvious.

Pesticide exposure alters follicle-stimulating hormone levels in Mexican agricultural workers.

Organophosphorous pesticides (OPs) are suspected of altering reproductive function by reducing brain acetylcholinesterase activity and monoamine levels, thus impairing hypothalamic and/or pituitary endocrine functions and gonadal processes. Our objective was to evaluate in a longitudinal study the association between OP exposure and serum levels of pituitary and sex hormones. Urinary OP metabolite levels were measured by gas-liquid chromatography, and serum pituitary and sex hormone levels by enzymatic immunoassay and radioimmunoassay in 64 men. A total of 147 urine and blood samples were analyzed for each parameter. More than 80% of the participants had at least one OP metabolite in their urine samples. The most frequent metabolite found was diethylthiophosphate (DETP; 55%), followed by diethylphosphate (DEP; 46%), dimethylthiophosphate (DMTP; 32%), and dimethyldithiophosphate (DMDTP; 31%). However, the metabolites detected at higher concentrations were DMTP, DEP, DMDTP, and dimethylphosphate. There was a high proportion of individuals with follicle-stimulating hormone (FSH) concentrations outside the range of normality (48%). The average FSH serum levels were higher during the heavy pesticide spraying season. However, a multivariate analysis of data collected in all periods showed that serum FSH levels were negatively associated with urinary concentrations of both DMTP and DMDTP, whereas luteinizing hormone (LH) was negatively associated with DMTP. We observed no significant associations between estradiol or testosterone serum levels with OP metabolites. The hormonal disruption in agricultural workers presented here, together with results from experimental animal studies, suggests that OP exposure disrupts the hypothalamic-pituitary endocrine function and also indicates that FSH and LH are the hormones most affected.

Organophosphorothionate pesticides inhibit the bioactivation of imipramine by human hepatic cytochrome P450s.

The drug-toxicant interaction between the antidepressant imipramine (IMI) and three organophosphorothionate pesticides (OPTs), to which humans may be chronically and simultaneously exposed, has been investigated in vitro. Concentrations of IMI (2-400 microM) and OPTs (< or =10 microM) representative of actual human exposure have been tested with recombinant human CYPs and human liver microsomes (HLM). The different CYPs involved in IMI demethylation to the pharmacologically active metabolite desipramine (DES) were CYP2C19 > CYP1A2 > CYP3A4. The OPTs significantly inhibited (up to >80%) IMI bioactivation catalyzed by the recombinant CYPs tested, except CYP2D6, and by HLM; the inhibition was dose-dependent and started at low pesticide concentrations (0.25-2.5 microM). The OPTs, having lower K(m) values, efficiently competed with IMI for the enzyme active site, as in the case of CYP2C19. However, with CYP1A2 and CYP3A4, a time- and NADPH-dependent mechanism-based inactivation also occurred, consistently with irreversible inhibition due to the release of the sulfur atom, binding to the active CYP during OPT desulfuration. At low IMI and OPT concentrations, lower IC50 values have been obtained with recombinant CYP1A2 (0.7-1.1 microM) or with HLM rich in 1A2-related activity (2-10.8 microM). The K(i) values (2-14 microM), independent on substrate concentrations, were quite low and similar for the three pesticides. Exposure to OPTs during IMI therapeutic treatments may lead to decreased DES formation, resulting in high plasma levels of the parent drug, eventual impairment of its pharmacological action and possible onset of adverse drug reactions (ADRs).

[Peripheral neuropathy and deterioration of the cognitive functions associated to chronic exposure to organophosphates]. / Neuropatía periférica y deterioro de las funciones cognitivas asociados a exposición crónica a organofosforados.

OBJECTIVE: To analyze the relationship between the chronic exposure (EC) to organophosphated insecticides (IOF) with the presence of peripheral neuropathy (NP) and the deterioration of cognitive functions (DFC). DESIGN OF THE STUDY: Cross-section, comparative and observational. MATERIAL AND METHODS: Workers with more than three months of seniority, who participated in any stage of the process of production of IOF were included, and those with alterations in the neurological function were excluded. Two control groups were made up: the first one, without direct exposure to IOF, and the second one, without any exposure whatsoever to IOF. A physical examination was practiced on inferior limbs and a motor neuroconduction of the fibular nerves was carried out. The Minimum Exam of the Mental State was set (Mental Mini). The sampling was not random. RESULTS: It was found that the presence of DFC and the clinical data for NP were greater in the exposed group than in the controls, (p < 0.05). Besides, data with a tendency to the continuation in the parameters of latencies, widths and waves F for the motor neuroconduction of inferior limbs were presented. CONCLUSIONS: The long term exposure to IOF is related to deterioration in the cognitive function; as well as to the neurological manifestations in inferior limbs that condition an injury in the peripheral nerve, without being conclusive about the loss of its function.

Chromosomal aberrations in human lymphocytes exposed to the anticholinesterase pesticide isofenphos with mechanisms of leukemogenesis.

Human lymphocytes were exposed to the leukemogenic pesticide isofenphos (IFP) to investigate its effects on chromosomal DNA and cholinergic homeostasis using cholinesterase activity as a marker. Isolated peripheral lymphocytes were administered concentrations of IFP ranging from 0.1 ng/ml to 10 microg/ml. The absence (Group 1) and presence (Group 2) of DNA repair inhibitors 4 mM hydroxyurea (HU), 40 microM cytosine arabinoside (ARA-C) and an NADPH regenerating system (NRS) (Group 3) were analyzed at 1, 6 and 24 h by single cell gel electrophoresis using the comet assay. Significant damage to DNA directly from IFP at 1 h by remarkably low concentrations was observed in Group 1, escalating in Group 2 with DNA repair inhibition, while Group 3 disruptions were highest due to the presence of the NRS P-450 microsomal fraction conducive to producing reactive IFP-oxon and N-desalkyl metabolites. The extent of DNA aberrations increased further in parallel within the groups at 6 and 24 h. Male and female chemical sensitivities were similar on average (P < 0.01). Cholinesterase activity measured in a satellite group was inhibited with 0.1 microg/ml IFP by 69, 62, and 48% at 1, 6, and 24 h, respectively, indicating gradual induction of compensatory synthesis. Restoration of cholinergic homeostasis may be exceptionally impaired at higher IFP concentrations from acetyl-CoA depletion [Leuk. Res. 25 (2001) 883]. In summary, these studies reveal that exposure to the organophosphate pesticide isofenphos induces human DNA mutation beyond endogenous repair capacity and disrupts cholinergic nuclear signaling affectively constructing the mutator phenotype of leukemogenesis.

Inhalational exposure to nerve agents.

The respiratory system plays a major role in the pathogenesis of nerve agent toxicity. It is the major route of entry and absorption of nerve agent vapor, and respiratory failure is the most common cause of death follow-ing exposure. Respiratory symptoms are mediated by chemical irritation,muscarinic and nicotinic receptor overstimulation, and central nervous system effects. Recent attacks have demonstrated that most patients with an isolated vapor exposure developed respiratory symptoms almost immediately. Most patients had only mild and transient respiratory effects, and those that did develop significant respiratory compromise did so rapidly. These observations have significant ramifications on triage of patients in a mass-casualty situation, because patients with mild-to-moderate exposure to nerve agent vapor alone do not require decontamination and are less likely to develop progressive symptoms following initial antidote therapy. Limited data do not demonstrate significant long-term respiratory effects following nerve agent exposure and treatment. Provisions for effective respiratory protection against nerve agents is a vital consideration in any emergency preparedness or health care response plan against a chemical attack.

Pesticide patch test series for the assessment of allergic contact dermatitis among banana plantation workers in panama.

BACKGROUND: Irritant contact dermatitis and allergic contact dermatitis (ACD) are frequent among agricultural workers and require targeted interventions. Patch testing is necessary for differential diagnosis, but patch testing with pesticides is uncommon. OBJECTIVE: This study explores the frequency of ACD and sensitization to pesticides among highly exposed banana plantation workers. METHODS: Frequently and recently used pesticides on banana plantations in Divala, Panama, were documented. A pesticide patch test tray specific for this population was prepared. A structured interview was administered to 366 participants, followed by a complete skin examination. The pesticide patch test series, as well as a standard patch test series, was applied to 37 workers with dermatoses likely to be pesticide related and to 23 control workers without dermatoses. RESULTS: The pesticide patch tests identified 15 cases (41%) of ACD (20 positive reactions) among the 37 workers diagnosed with pesticide dermatosis. Three controls had allergic reactions to pesticides (4 positive reactions). The pesticides were carbaryl (5 cases), benomyl (4 cases), ethoprophos (3), chlorothalonil (2), imazalil (2), glyphosate (2), thiabendazole (2), chlorpyrifos (1), oxyfluorfen (1), propiconazole (1), and tridemorph (1). Ethoprophos and tridemorph had not been previously identified as sensitizers. Thus, the prevalence of ACD was 0.03 (15 of 366). On the basis of observed prevalences of positive patch-test reactions among the subgroups with and without dermatoses, we estimated that > or = 16% of the entire population may be sensitized to pesticides. CONCLUSION: Sensitization to pesticides among banana plantation workers is a frequent occupational health problem. Pesticide patch test trays should be used in assessing skin diseases in highly exposed workers.

Effect of edifenphos on compensatory ovarian hypertrophy, follicular kinetics and estrous cycle in hemicastrated rats.

Edifenphos, an organophosphate fungicide, was administered to hemicastrated albino rats intraperitoneally for 15 consecutive days in graded doses of 2 to 8 mg/kg body weight. In the oil-treated hemicastrated control group, ovarian weight and total number of healthy and atretic follicles were significantly higher than the same parameters in sham-operated control animals, and the estrous cycle was normal. Treatment with 2, 4, 6, or 8 mg/kg/d edifenphos significantly decreased ovarian weight (-6.75, -9.79, -18.71, and -34.13 percent, respectively) below that of the controls. Although treatment with 2 or 4 mg/kg/d edifenphos did not change the number of healthy and atretic follicles, a dosage of 4 mg/kg/d significantly decreased the number and duration of estrous cycles. Treatment with 6 or 8 mg/kg/d, however, significantly decreased the number and duration of different phases of the estrous cycle as well. In all treated groups, the weights of the thymus and uterus were significantly reduced when compared with those of hemicastrated oil-treated control animals.

Relationship between biomarker activity and developmental endpoints in Chironomus riparius Meigen exposed to an organophosphate insecticide.

The biomarkers acetylcholinesterase (AChE) and glutathione S-transferase (GST) were measured in fourth-instar Chironomus riparius Meigen larvae exposed to the organophosphate insecticide pirimiphos methyl (0, 5, 10, and 50ng/g) for 48 or 96h, and at high or low food ration. Larvae exposed to 50ng/g pirimiphos methyl died within 48h. The weight of larvae exposed to 10ng/g pirimiphos methyl was significantly lower than those exposed to 0 and 5ng/g. AChE activity was significantly reduced in larvae exposed to 10ng/g, but GST activity remained unaffected. Lower food ration reduced larval weights across all treatments but did not affect biomarker measurements. Insecticide exposure was associated with a longer time to adult emergence and oviposition, fewer egg masses, a greater proportion of deformed egg masses, and fewer eggs.