First biochemical and behavioural analysis of the response of the scorpion Urophonius brachycentrus (Thorell: 1876) upon exposure to an organophosphate.

Scorpionism is an increasing public health problem in the world. Although no specific methodology or product is currently available for the control of those arachnids, the use of insecticides could be an effective tool. Chlorpyrifos is one of the insecticides used, but to date, whether scorpions recognise surfaces with that insecticide and how it affects their physiology and/or biochemistry is unknown. In the present study, we observed that scorpions recognise surfaces with 0.51 and 8.59 µg/cm2 of chlorpyrifos and avoid those areas. The 0.51 µg/cm2 concentration produced a decrease in acetylcholinesterase and an increase in catalase, superoxide dismutase and glutathione S-transferase, whereas the 8.59 µg/cm2 concentration evoked a decrease in acetylcholinesterase and an increase in catalase and glutathione S-transferase. Using the comet assay, we observed that the insecticide at 0.17, 0.51 and 8.59 µg/cm2 caused DNA damage. Finally, we found that the insecticide does not generate significant variations in glutathione peroxidase, glutathione reductase, the amount of protein or lipid peroxidation. The present results offer a comprehensive understanding of how scorpions respond, both at the biochemical and behavioural levels, when exposed to insecticides.

Status of Acaricide Resistance and Detecting the Knockdown Resistance Mutation T2134A in the Cattle Tick Rhipicephalus microplus (Acari: Ixodidae) from Northeastern Mexico.

Rhipicephalus microplus is the most important tick in veterinary medicine, given its repercussions on animal production. The principal strategy to avoid adverse effects associated with R. microplus is the chemical control of tick populations through organosynthetic acaricides. Therefore, monitoring susceptibility to acaricides is paramount in any control program. This study aimed to analyze the resistance status of 2 populations of R. microplus from northeastern Mexico to the organochlorine (OC) lindane, organophosphates (OP) coumaphos, chlorfenvinphos, diazinon, and chlorpyrifos, and the synthetic pyrethroids (SPs) flumethrin, deltamethrin, and cypermethrin. Discriminating doses (DD) of each acaricide were used in the larval packet bioassay (LPT). Additionally, the presence of the knockdown resistance (kdr) mutation T2134A associated with pyrethroid resistance was evaluated using allele-specific polymerase chain reaction (PCR). The populations of R. microplus showed a high frequency of resistance to SP, with mortality rates of less than 5%; they also showed resistance to the OPs (diazinon and chlorpyrifos) with mortality rates ranging from 1.29% to 34.62%; meanwhile, they were susceptible to coumaphos and chlorfenvinphos. Mortality rates higher than 66% were observed for lindane, indicating susceptibility. The mutant allele of the kdr mutation T2134A was detected in 75% and 100% of the pools analyzed. The populations studied presented a highly resistant profile to pyrethroids, with the presence of the kdr mutant allele A2134. The susceptibility to the organophosphates such as coumaphos and chlorfenvinphos of R. microplus from northeastern Mexico should be noted.

Thirty-minute test to detect insecticide resistance under field conditions: the case of the Asian citrus psyllid.

BACKGROUND: Diaphorina citri Kuwayama is one of the most destructive citrus pests worldwide. It is controlled mainly through applications of conventional insecticides. Methodologies used to estimate resistance to insecticides do not correlate with field efficacy, and do not provide timely and reliable information to make decisions at a site where spraying is needed. The use of diagnostic doses with 30-min exposure is proposed for estimating the resistance of D. citri to imidacloprid, spinosad, malathion and chlorpyrifos at the orchard level. RESULTS: Under laboratory conditions, we estimated the lowest doses that caused 100% mortality within 30 min of exposure (diagnostic dose) in a susceptible D. citri colony. The diagnostic doses for imidacloprid, spinosad, malathion and chlorpyrifos were 7.4, 4.2, 1.0 and 5.5 mg a.i. L-1 , respectively. Under field conditions, we applied the diagnostic doses to D. citri feeding on Citrus aurantifolia Swingle at five localities in Michoacan state, Mexico (Nueva Italia, Santo Domingo, El Varal, Gambara and El Ceñidor). Additionally, the field efficacy of these insecticides against these populations was evaluated. A significant correlation between field efficacy and mortality was observed with the diagnostic doses for imidacloprid, malathion and chlorpyrifos (R2 ≥ 0.93). The correlation for spinosad could not be estimated because the mortality caused by the diagnostic dose and its field efficacy at all study sites was consistently >98%. CONCLUSIONS: Field efficacy and resistance were estimated based on the field diagnostic doses with 30-min exposure for all tested insecticides. Consequently, growers and pest management technicians can estimate the performance of the evaluated insecticides at the orchard level and before insecticide application. © 2023 Society of Chemical Industry.

Susceptibility monitoring and comparative gene expression of susceptible and resistant strains of Spodoptera frugiperda to lambda-cyhalothrin and chlorpyrifos.

BACKGROUND: Spodoptera frugiperda (J. E. Smith) is a widespread agricultural pest with several records of resistance to different insecticides and Bt proteins, including the neurotoxic insecticides chlorpyrifos (organophosphate) and lambda-cyhalothrin (pyrethroid). Here, we (i) characterized and monitored the susceptibility of field populations of S. frugiperda to chlorpyrifos (194 populations) and lambda-cyhalothrin (197 populations) collected from major maize-growing regions of Brazil from 2003 to 2016, and (ii) compared gene expression levels of laboratory-selected, chlorpyrifos- and lambda-cyhalothrin-resistant strains to a susceptible reference strain (Sf-ss) of S. frugiperda. RESULTS: The susceptibility monitoring detected average survival ranging from 29.3% to 36.0% for chlorpyrifos, and 23.1% to 68.0% for lambda-cyhalothrin. The resistance ratio of the chlorpyrifos-resistant strain (Clo-rr) was 25.4-fold and of the lambda-cyhalothrin-resistant strain (Lam-rr) was 21.5-fold. We identified 1098 differentially expressed genes (DEGs) between Clo-rr and Sf-ss, and 303 DEGs between Lam-rr and Sf-ss. Functional analyses of the DEGs revealed the up-regulation of several detoxification enzymes, mainly cytochrome P450 belonging to CYP3 and CYP6 clans. Genes associated with regulatory processes, such as the forkhead box class O (FoxO) transcription factor were also up-regulated. Variant analysis of target-site mutations for both pesticides identified the A201S and F290V mutations in acetylcholinesterase-1, both occurring in heterozigosis in the Clo-rr S. frugiperda strain. CONCLUSION: Our data show that the overexpression of the enzymatic detoxification machinery is the main difference to explain the resistance of Clo-rr and Lam-rr strains of S. frugiperda to chlorpyrifos and lambda-cyhalothrin, although a target-site mutation also contributes to the Clo-rr resistance to chlorpyrifos. © 2023 Society of Chemical Industry.

Chlorpyrifos stimulates ABCC-mediated transport in the intestine of the rainbow trout Oncorhynchus mykiss.

The organophosphorus pesticide chlorpyrifos, detected in water and food worldwide, has also been found in the Río Negro and Neuquén Valley, North Patagonia, Argentina, where the rainbow trout, Oncorhynchus mykiss, is one of the most abundant fish species. We analyzed whether chlorpyrifos affects the transport activity of the ATP-binding cassette protein transporters from the subfamily C (ABCC), which are critical components of multixenobiotic resistance. We exposed ex vivo O. mykiss middle intestine strips (non-polarized) and segments (polarized) for one hour to 0 (solvent control), 3, 10, and 20 µg L-1 and to 0, 10, and 20 µg L-1 chlorpyrifos, respectively. We estimated the Abcc-mediated transport rate by measuring the transport rate of the specific Abcc substrate 2,4-dinitrophenyl-S-glutathione (DNP-SG). In addition, we measured the enzymatic activity of cholinesterase, carboxylesterase, glutathione-S-transferase, and 7-ethoxyresorufin-O-deethylase (EROD, indicative of the activity of cytochrome P450 monooxygenase 1A, CYP1A). We also measured lipid peroxidation using the thiobarbituric acid reactive substances method and the gene expression of Abcc2 and genes of the AhR pathway, AhR, ARNT, and cyp1a, by qRT-PCR. Chlorpyrifos induced the DNP-SG transport rate in middle intestine strips in a concentration-dependent manner (49-71%). In polarized preparations, the induction of the DNP-SG transport rate was observed only in everted segments exposed to 20 µg L-1 chlorpyrifos (40%), indicating that CPF only stimulated the apical (luminal) transport flux. Exposure to chlorpyrifos increased GST activity by 42% in intestine strips and inhibited EROD activity (47.5%). In addition, chlorpyrifos exposure inhibited cholinesterase (34-55%) and carboxylesterase (33-42.5%) activities at all the concentrations assayed and increased TBARS levels in a concentration-dependent manner (71-123%). Exposure to 20 µgL-1 chlorpyrifos did not affect the mRNA expression of the studied genes. The lack of inhibition of DNP-SG transport suggests that chlorpyrifos is not an Abcc substrate. Instead, CPF induces the activity of Abcc proteins in the apical membrane of enterocytes, likely through a post-translational pathway.

Determination of hormoligosis of organophosphate insecticides against Phenacoccus solenopsis.

Cotton mealybug is a highly invasive pest of agricultural crops worldwide. Major agriculturists most rely on the use of insecticides for the control of pesticides. So, the indiscriminate use of insecticides leads to resistance development in recent years. For this purpose, an experiment was conducted using different concentrations of the three insecticides (profenfos chlorpyrifos and triazophos) to check the hormoligosis effects against cotton mealybug (CMB) in laboratory conditions. Investigation of variations for % mortality of adults of CMB after three days revealed that all treatments had statistically significant (P ˂ 0.05). The highest mortality was observed at the highest concentrations of profenofos 2.4% (38.55%). After 7 days, all the treatments were significant with difference in means (P ˂ 0.05). The highest mortality was recorded at the highest dilution of pesticide profenofos 2.4% (77.11%). The values of fecundity and longevity exposed a valid difference among treatments (P ˂ 0.05). Maximum fecundity was observed at the concentration 2.4% (181.41%) and longevity showed (38.46%). The highest mortality was observed at a concentration of triazophos 4% (27.98%). For chlorpyriphos the highest mortality was examined at concentration 4% (24.79%). The fecundity showed a statistically significant difference for different concentrations of triazophos and chlorpyriphos (P ˂ 0.05). The results of the recent study provide valuable information regarding the selection of insecticides and hormoligosis effects. The study can be helpful in the implications of integrated pest management of P. solenopsis.

Compatibility of different Metarhizium spp. propagules with synthetic acaricides for controlling Rhipicephalus microplus.

The inappropriate use of synthetic acaricides has selected resistant Rhipicephalus microplus populations. The present study evaluated the compatibility of different Metarhizium spp. propagules (conidia, blastospores, and microsclerotia) by incubating them with synthetic acaricides (amitraz, deltamethrin, and a combination of cypermethrin, chlorpyrifos, and citronellal) for 1 h, 5 h, 10 h, and 24 h. Conidia and microsclerotia of the tested isolates were usually more tolerant to synthetic acaricides than blastospores. Our study also analyzed the in vitro effect of deltamethrin associated with fungal propagules for controlling a population of R. microplus females that were not susceptible to this synthetic acaricide. The use of entomopathogenic fungi in association with deltamethrin in this tick population caused a greater tick control than did the use of the fungus or the synthetic acaricide separately.

Field-evolved resistance to chlorpyrifos by Spodoptera frugiperda (Lepidoptera: Noctuidae): Inheritance mode, cross-resistance patterns, and synergism.

BACKGROUND: Fall armyworm (FAW), Spodoptera frugiperda (Smith), is an economically important pest worldwide. In this study, we selected a genotype of FAW resistant to chlorpyrifos from a field-collected population, characterized the genetic basis of resistance, and evaluated cross-resistance and mechanisms of resistance using synergists. RESULTS: The LD50 values of chlorpyrifos for the resistant (Clorp-R) and susceptible (Sus) FAW genotypes were 24.26 and 0.023 µg per larva, respectively, representing a resistance ratio > 1050-fold. The LD50 values of chlorpyrifos against heterozygotes were 3.34 and 4.00 µg per larva, suggesting that resistance is autosomally inherited. The chlorpyrifos resistance in FAW was influenced by few genes, with the minimum numbers of segregations being 1.74 and 1.88. On chlorpyrifos-sprayed plants and leaves, Clorp-R and heterozygote genotypes showed >95% and >52% survival, respectively, whereas the Sus genotype had no survival, indicating that the resistance is incompletely dominant at the field rate of chlorpyrifos. The Clorp-R genotype presented some cross-resistance to acephate, but low cross-resistance to thiodicarb, methomyl, chlorfenapyr, flubendiamide, methoxyfenozide, spinetoram, and teflubenzuron. The synergists piperonyl butoxide, diethyl maleate, and S,S,S-tributyl phosphorotrithiotate did not have relevant effects on the Clorp-R genotype, suggesting a minor role for metabolic resistance. CONCLUSIONS: The inheritance of chlorpyrifos resistance in FAW was characterized as autosomal, incompletely dominant, and polygenic, with metabolic resistance playing a small role in the detoxification of chlorpyrifos. Low cross-resistance between chlorpyrifos and other mode of action (MoA) insecticides occurs in FAW, highlighting the importance of considering the rotation of MoA as a strategy to delay resistance. © 2021 Society of Chemical Industry.

Effects of chlorpyrifos on growth and aflatoxin B1 production by Aspergillus section Flavi strains on maize-based medium and maize grains.

Chlorpyrifos is one of the most used insecticides in agro-ecosystems and is repeatedly applied due to the increase in pest resistance, which leads to environmental accumulation. The aim of this work was to evaluate the effect of chlorpyrifos on growth and aflatoxin B1 (AFB1) production by four Aspergillus section Flavi strains, under different water conditions-aW (0.93, 0.95 and 0.98)-on maize-based medium (MMEA) and maize grains supplied with 0.06 to 1.4 mmol/L of chlorpyrifos. MMEA plates were incubated at 18, 28, and 37 °C and plates with maize grains at 25 °C during 21 days. Chlorpyrifos stimulated the growth and AFB1 production of non-target organisms, such as Aspergillus section Flavi strains, both at low (0.06 mmol/L) and at high concentrations (1.4 mmol/L) on MMEA and maize grains. Stimulation occurred over a wide range of temperature and aw conditions. The toxin concentration produced by the two strains on MMEA at 18 °C increased when the concentration of chlorpyrifos also increased, being most significant at 0.6 mmol/L. In conclusion, the presence of chlorpyrifos should be considered as a factor, together with environmental conditions, for the development of effective production practices of maize grains, in order to avoid fungal growth and AFB1 production, to prevent both economic losses and risks to human and animal health.

Glutathione in Chlorpyrifos-and Chlorpyrifos-Oxon-Induced Toxicity: a Comparative Study Focused on Non-cholinergic Toxicity in HT22 Cells.

Chlorpyrifos (CPF) is a neurotoxic organophosphorus (OP) insecticide widely used for agricultural purposes. CPF-mediated neurotoxicity is mainly associated with its anticholinesterase activity, which may lead to a cholinergic syndrome. CPF metabolism generates chlorpyrifos-oxon (CPF-O), which possesses higher anticholinesterase activity and, consequently, plays a major role in the cholinergic syndrome observed after CPF poisoning. Recent lines of evidence have also reported non-cholinergic endpoints of CPF- and CPF-O-induced neurotoxicities, but comparisons on the non-cholinergic toxic properties of CPF and CPF-O are lacking. In this study, we compared the non-cholinergic toxicities displayed by CPF and CPF-O in cultured neuronal cells, with a particular emphasis on their pro-oxidant properties. Using immortalized cells derived from mouse hippocampus (HT22 line, which does present detectable acetylcholinesterase activity), we observed that CPF-O was 5-fold more potent in decreasing cell viability compared with CPF. Atropine, a muscarinic acetylcholine receptor antagonist, protected against acetylcholine (ACh)-induced toxicity but failed to prevent the CPF- and CPF-O-induced cytotoxicities in HT22 cells. CPF or CPF-O exposures significantly decreased the levels of the antioxidant glutathione (GSH); this event preceded the significant decrease in cell viability. Pretreatment with N-acetylcysteine (NAC, a GSH precursor) protected against the cytotoxicity induced by both CPF and CPF-O. The present study indicates that GSH depletion is a non-cholinergic event involved in CPF and CPF-O toxicities. The study also shows that in addition of being a more potent AChE inhibitor, CPF-O is also a more potent pro-oxidant molecule when compared with CPF, highlighting the role of CPF metabolism (bioactivation to CPF-O) in the ensuing non-cholinergic toxicity.

Resistance profile and molecular characterization of pyrethroid resistance in a Rhipicephalus microplus strain from Colombia.

Intensive use of chemical acaricides for the control of cattle ticks (Rhipicephalus microplus) has led to the development of multiple acaricide resistance in Colombia. The present study aimed to characterize, using toxicological bioassays and molecular biology techniques, the resistance profile of a tick strain isolated from the Arauca state, Northeast Colombia. Commercial acaricides were used in adult immersion tests to determine its in vitro efficacies. Deltamethrin showed very low activity (4-7.3%), a mixture of cypermethrin and chlorpyrifos had intermediate efficacy (64-75.2%), and ethion presented the highest activity (88.5-100%). A colony (Arauquita strain) was established and larval immersion tests confirmed high resistance level to deltamethrin (241-fold) and susceptibility to ivermectin. A quantitative polymerase chain reaction-high resolution melt technique was used to identify single nucleotide polymorphisms (SNPs) in the para-sodium channel gene. All of the genotyped individuals were mutant, presenting one (n = 7), two (n = 7) or three (n = 9) SNPs previously associated with pyrethroid resistance. Sequencing revealed a novel mutation (F712L), that was found for the first time in R. microplus ticks from South America. This is the first description of mutations associated with pyrethroid resistance in R. microplus from Colombia. The acaricide resistance pattern found in the Arauquita strain is similar to other parts of Colombia.

Development and removal ability of non-toxigenic Aspergillus section Flavi in presence of atrazine, chlorpyrifos and endosulfan.

This study evaluated the in vitro effect of three concentrations of atrazine, chlorpyrifos and endosulfan on the growth parameters of four non-toxigenic Aspergillus section Flavi strains. The ability of the strains to remove these pesticides in a synthetic medium was also determined. Growth parameters were measured on soil extract solid medium supplied with 5, 10 and 20mg/l of each pesticide, and conditioned to -0.70, -2.78, -7.06 and -10.0 water potential (MPa). Removal assays were performed in Czapek Doc medium (CZD) supplied with 20mg/l of each pesticide under optimal environmental conditions (-2.78 of MPa and 25°C). The residual levels of each pesticide were detected by the reversed-phase HPLC/fluorescence detection system. The lag phases of the strains significantly decreased in the presence of the pesticides with respect to the control media. This result indicates a fast adaptation to the conditions assayed. Similarly, the mycelial growth rates in the different treatments increased depending on pesticide concentrations. Aspergillus oryzae AM 1 and AM 2 strains showed high percentages of atrazine degradation (above 90%), followed by endosulfan (56 and 76%) and chlorpyrifos (50 and 73%) after 30 days of incubation. A significant (p<0.001) correlation (r=0.974) between removal percentages and growth rate was found. This study shows that non-toxigenic Aspergillus section Flavi strains from agricultural soils are able to effectively grow in the presence of high concentrations of atrazine, chlorpyrifos and endosulfan under a wide range of MPa conditions. Moreover, these strains have the ability to remove high levels of these pesticides in vitro in a short time.

Effect of exposition to chlorpyrifos upon plasmatic cholinesterases, hematology and blood biochemistry values in Bothrops asper (Serpentes: Viperidae).

The terciopelo (Bothrops asper), is one of the most important venomous snakes in Costa Rica and common on agriculture where insecticides are frequently used for pest control. To assess the exposure to organophosphates on captive B. asper, an experiment using chlorpyrifos and butyrylcholinesterase (BChE), as a biomarker was conducted. In addition to BChE, hematology, aspartate aminotransferase (AST), total proteins (TP) and albumin were measured after exposure. Different concentrations of chlorpyrifos were used in Group A (0.1%) and B (1%), while the Control Group received distilled water; each group was composed of 5 snakes. Values of BChE, AST, TP, and albumin were measured before exposure, and at 6, 12, 24, 196, 360 and 528 h post-exposure. Hematology values were measured after 24 h post-exposure. As result, an important variation between subjects in all groups before exposure was obtained. Moreover, BChE activity showed 37% inhibition of Group A when compared to Control Group at 12 h post-exposure, and a higher inhibition of Group B (97%) related to Control Group, at 6 h post-exposure. Recovery of BChE occurred towards 528 h, never reaching initial values. Despite some variation in the rest of parameters used, a marked relative lymphopenia and monocytosis occurred at 24 h, assuming stress as the main cause.

Insecticide resistance of adults and nymphs of Asian citrus psyllid populations from Apatzingán Valley, Mexico.

BACKGROUND: Control of the Asian citrus psyllid Diaphorina citri Kuwayama, the most important pest of citrus worldwide, is based on the use of insecticides, though unsatisfactory results have recently been reported. In this study, insecticide resistance of D. citri to three insecticides (bifenthrin, malathion, and chlorpyrifos) was examined. RESULTS: Three populations (designated Dci-CParácuaro, Dci-El Junco, and Dci-Antúnez) of both adults and fourth-instar D. citri individuals were collected in 2014 at two different times and on one occasion, respectively, from three locations (Crucero de Parácuaro, El Junco, and Antúnez). These locations represent the major commercial Mexican lemon production areas in the Apatzingán Valley in the state of Michoacán, Mexico. The three populations of D. citri adults and fourth-instar nymphs at the different collection times showed low levels of resistance (≤7-fold) to bifenthrin, but were very resistant to malathion (≤345- and ≤432-fold for adults and fourth instars, respectively) and chlorpyrifos (≤2435- and ≤1424-fold for adults and fourth instars, respectively). CONCLUSION: Resistance levels to the tested insecticides were highly variable but homogeneous among seasons and localities. Resistance management programmes that include crop sanitation, use of biological and cultural control practices, and rotation of insecticide classes should be established, particularly in areas where D. citri has developed resistance to malathion and chlorpyrifos. © 2017 Society of Chemical Industry.

The impact of six insecticides commonly used in control of agricultural pests on the generalist predator Hippodamia convergens (Coleoptera: Coccinellidae).

Hippodamia convergens is an important predator found in different agroecosystems. We evaluated the impacts of six insecticides on eggs, larvae and adults of this predator. For eggs, all insecticides reduced larval hatching rates, but did not affect egg duration. Chlorpyrifos and phosmet reduced larval survival; and chlorpyrifos, etofenprox and phosmet prolonged the larva development time. The survival and duration of pupae were not affected by all insecticides tested. Chlorpyrifos reduced fecundity, fertility and longevity when eggs were sprayed. For first-instar larvae, chlorpyrifos, etofenprox, phosmet and imidacloprid caused 100% mortality, while azadirachtin and thiamethoxam caused 35.0 and 52.7% mortality, respectively. However, azadirachtin and thiamethoxam did not affect the other biological parameters of the predator. In adults, chlorpyrifos, etofenprox and phosmet reduced adult survival. Chlorpyrifos, etofenprox, and phosmet reduced fecundity and longevity, but did not affect fertility. Azadirachtin, imidacloprid and thiamethoxam did not affect fecundity, fertility or longevity. Based on demographic parameters, all insecticides reduced the net reproductive rate (Ro), intrinsic rate of increase (r) and finite rate of increase (λ) of the predator when eggs were treated directly. Azadirachtin, chlorpyrifos, etofenprox and phosmet increased the mean generation time (T), while the effects of imidacloprid and thiamethoxam were similar to the control. When first-instar larvae were treated, azadirachtin and thiamethoxam reduced the Ro, r and λ. Thiamethoxam increased the T value, while the effects of the other insecticides were similar to the control. These insecticides should be used with caution, in order to reduce their harmful effects on the predator in agroecosystems.

Neurobehavioral and biochemical changes in Nauphoeta cinerea following dietary exposure to chlorpyrifos.

The present study aimed to increase our understanding about the mode of toxic action of organophosphate pesticides in insects by evaluating the biochemical and neurobehavioral characteristics in Nauphoeta cinerea exposed to chlorpyrifos (CPF)-contaminated diet. The insects were exposed for 35 consecutive days to CPF at 0.078, 0.15625, 0.3125 and 0.625µg/g feed. Locomotor behavior was assessed for a 10-min trial in a novel arena and subsequently, biochemical analyses were carried out using the cockroaches' heads. In comparison to control, CPF-exposed cockroaches showed significant decreases in the total distance traveled, body rotation, turn angle and meandering, along with significant increase in the number of falls, time and episodes of immobility. The marked decrease in the exploratory profiles of CPF-exposed cockroaches was confirmed by track plots, whereas occupancy plot analyses showed a progressive dispersion at 0.15625µg/g feed group. Moreover, the heads of CPF-exposed cockroaches showed marked decrease in acetylcholinesterase activity and antioxidant status with concomitant significant elevation in dichlorofluorescein oxidation and lipid peroxidation levels in CPF-treated cockroaches. Gas Chromatography-Mass Spectrometry analyses revealed bioaccumulation of CPF in cockroaches exposed to concentrations above 0.078µg/g feed. The findings from this investigation showed N. cinerea as a value model organism for the risk assessment of environmental organophosphate contamination in insects.

Integrated control of an acaricide-resistant strain of the cattle tick Rhipicephalus microplus by applying Metarhizium anisopliae associated with cypermethrin and chlorpyriphos under field conditions.

The efficacy of the fungus Metarhizium anisopliae to control ticks has been shown in several in vitro experiments. However, few studies have been undertaken in field conditions in order to demonstrate the applicability of its use as a biological control of ticks and its combination with chemical acaricides. The aim of the present study was to evaluate the efficacy of M. anisopliae to control an acaricide-resistant strain of Rhipicephalus microplus under laboratory and field conditions. First, the compatibility of M. anisopliae strain (TIS-BR03) with commercial acaricides and its potential to control the cattle tick were evaluated in vitro. In general, acaricide treatments had mild effects on fungus viability. In the field experiment, the median of treatment efficacy with acaricide only, M. anisopliae only and combination of M. anisopliae with acaricide were 71.1%, 56.3% and 97.9%, respectively. There is no statistical difference between groups treated with M. anisopliae and acaricide alone. Thus, in this work we have demonstrated the applicability of M. anisopliae use associated or not with chemical acaricides on field conditions in order to control an acaricide-resistant strain of the cattle tick R. microplus.

Effect of a spray formulation on the reproductive parameters of a susceptible population of Rhipicephalus (Boophilus) microplus.

The number of studies emphasizing the possible damage that acaricidal spray formulations can cause on engorged female ticks' reproductive parameters is small. The present study evaluated the deleterious effects of a spray formulation (dichlorvos 60% + chlorpyrifos 20%) on the reproductive parameters of a susceptible population of Rhipicephalus (B.) microplus females, using the Stall Test. The ticks were allocated randomly to treatments according to the mean numbers of females detached from each cow on days -3, -2 and -1 and the cattle pen location. The numbers of engorged female ticks that naturally detached from the cattle were counted daily from day 1 to day 30. For each group, 20 detached engorged female ticks or the available number collected daily were evaluated regarding reproductive parameters. Associations of organophosphates demonstrated elevated acaricidal efficacy, as well as deleterious effects on the reproductive parameters of R. (B.) microplus females. The engorged female weight (days 1 to 7), weight of egg masses (days 5 to 10) and larval hatching percentage (days 5 to 19) were decreased (P ≤ 0.05). It is possible that a formulation can lead to deleterious effects on R. (B.) microplus females when the tick population analyzed shows elevated sensitivity towards a particular formulation. However, further studies need to be conducted.

Susceptibility to chlorpyrifos in pyrethroid-resistant populations of Aedes aegypti (Diptera: Culicidae) from Mexico.

Resistance to the organophosphate insecticide chlorpyrifos was evaluated in females from six strains of Aedes aegypti (L.) that expressed high levels of cross-resistance to eight pyrethroid insecticides. Relative to LC50 and LC90 at 24 h of a susceptible New Orleans (NO) strain, three strains were highly resistant to chlorpyrifos (Coatzacoalcos, resistance ratio [RRLC90 = 11.97; Pozarica, RRLC90 = 12.98; and Cosoleacaque, RRLC50 = 13.94 and RRLC90 = 17.57), one strain was moderately resistant (Veracruz, RRLC90 = 5.92), and two strains were susceptible (Tantoyuca and Martinez de la Torre, RRLC50 and RRLC90 < 5) in bottle bioassays according to Centers for Disease Control and Prevention. Furthermore, high levels of alpha- or beta-esterase activity in the sample populations were correlated with resistance, suggesting that esterase activity may be a mechanism causing the development of organophosphate resistance in these populations. Overall, the populations in this study were less resistant to chlorpyrifos than to pyrethroids. Rotation of insecticides used in control activities is recommended to delay or minimize the occurrence of high levels of resistance to chlorpyrifos among local populations of Ae. aegypti. The diagnostic dose and diagnostic time for chlorpyrifos resistance monitoring was determined to be 85 microg per bottle and 30 min, respectively, using the susceptible NO strain.

Binary mixtures of azinphos-methyl oxon and chlorpyrifos oxon produce in vitro synergistic cholinesterase inhibition in Planorbarius corneus.

In this study, the cholinesterase (ChE) and carboxylesterase (CES) activities present in whole organism homogenates from Planorbarius corneus and their in vitro sensitivity to organophosphorous (OP) pesticides were studied. Firstly, a characterization of ChE and CES activities using different substrates and selective inhibitors was performed. Secondly, the effects of azinphos-methyl oxon (AZM-oxon) and chlorpyrifos oxon (CPF-oxon), the active oxygen analogs of the OP insecticides AZM and CPF, on ChE and CES activities were evaluated. Finally, it was analyzed whether binary mixtures of the pesticide oxons cause additive, antagonistic or synergistic ChE inhibition in P. corneus homogenates. The results showed that the extracts of P. corneus preferentially hydrolyzed acetylthiocholine (AcSCh) over propionylthiocholine (PrSCh) and butyrylthiocholine (BuSCh). Besides, AcSCh hydrolyzing activity was inhibited by low concentrations of BW284c51, a selective inhibitor of AChE activity, and also by high concentrations of substrate. These facts suggest the presence of a typical AChE activity in this species. However, the different dose-response curves observed with BW284c51 when using PrSCh or BuSCh instead of AcSCh suggest the presence of at least another ChE activity. This would probably correspond to an atypical BuChE. Regarding CES activity, the highest specific activity was obtained when using 2-naphthyl acetate (2-NA), followed by 1-naphthyl acetate (1-NA); p-nitrophenyl acetate (p-NPA), and p-nitrophenyl butyrate (p-NPB). The comparison of the IC(50) values revealed that, regardless of the substrate used, CES activity was approximately one order of magnitude more sensitive to AZM-oxon than ChE activity. Although ChE activity was very sensitive to CPF-oxon, CES activity measured with 1-NA, 2-NA, and p-NPA was poorly inhibited by this pesticide. In contrast, CES activity measured with p-NPB was equally sensitive to CPF-oxon than ChE activity. Several specific binary combinations of AZM-oxon and CPF-oxon caused a synergistic effect on the ChE inhibition in P. corneus homogenates. The degree of synergism tended to increase as the ratio of AZM-oxon to CPF-oxon decreased. These results suggest that synergism is likely to occur in P. corneus snails exposed in vivo to binary mixtures of the OPs AZM and CPF.