Combined toxicity and adverse outcome pathways of common pesticides on Chlorella pyrenoidosa.

Pesticides due to their extensive use have entered the soil and water environment through various pathways, causing great harm to the environment. Herbicides and insecticides are common pesticides with long-term biological toxicity and bioaccumulation, which can harm the human body. The concept of the adverse outcome pathway (AOP) involves systematically analyzing the response levels of chemical mixtures to health-related indicators at the molecular and cellular levels. The AOP correlates the structures of chemical pollutants, toxic molecular initiation events and adverse outcomes of biological toxicity, providing a new model for toxicity testing, prediction, and evaluation of pollutants. Therefore, typical pesticides including diquat (DIQ), cyanazine (CYA), dipterex (DIP), propoxur (PRO), and oxamyl (OXA) were selected as research objects to explore the combined toxicity of typical pesticides on Chlorella pyrenoidosa (C. pyrenoidosa) and their adverse outcome pathways (AOPs). The mixture systems of pesticides were designed by the direct equipartition ray (EquRay) method and uniform design ray (UD-Ray) method. The toxic effects of single pesticides and their mixtures were systematically investigated by the time-dependent microplate toxicity analysis (t-MTA) method. The interactions of their mixtures were analyzed by the concentration addition model (CA) and the deviation from the CA model (dCA). The toxicity data showed a good concentration-effect relationship; the toxicities of five pesticides were different and the order was CYA > DIQ > OXA > PRO > DIP. Binary, ternary and quaternary mixture systems exhibited antagonism, while quinary mixture systems exhibited an additive effect. The AOP of pesticides showed that an excessive accumulation of peroxide in green algae cells led to a decline in stress resistance, inhibition of the synthesis of chlorophyll and protein in algal cells, destruction of the cellular structure, and eventually led to algal cell death.

[Surveillance on dengue vector Aedes albopictus in Ningbo City in 2021].

OBJECTIVE: To analyze the density, distribution and insecticide resistance of Aedes albopictus in Ningbo City in 2021, so as to provide insights into formulation of dengue fever control strategies. METHODS: Four administrative villages were randomly selected from each county (district) in Ningbo City from April to November, 2021, to investigate the indoor population density of Aedes larvae, and the Breteau index (BI) was calculated. The population density of adult mosquitoes was investigated in residential areas, parks/bamboo forests, waste tire stacking sites/waste stations/construction sites in each county (district). On June 2021, larvae of the natural strain A. albopictus were collected from epidemic sites of dengue fever in Ningbo City in 2018, and raised in laboratory. Then, larvae and female mosquitoes without blood feeding were selected for insecticide resistance bioassays, while insecticide-sensitive strains of A. albopictus served as controls. The resistance of A. albopictus larvae to deltamethrin, beta-cypermethrin, propoxur, temephos and dichlorvos using the impregnation method, and the medium lethal concentration (LC50) and resistance ratio (RR) were calculated. The resistance of adult A. albopictus to beta-cypermethrin, permethrin, deltamethrin, propoxur and malathion was determined using the tube bioassay, and the mosquito mortality was calculated. RESULTS: A total of 10 072 small water containers from 9 935 households were investigated in Ningbo City in 2021, and there were 1 276 containers with Aedes larvae detected, with an average BI of 12.89. Totally 1 422 mosquito nets were allocated and 954 female A. albopictus were captured, with an average net trapping index of 1.34 mosquitoes/(net·hour). Both larval and adult A. albopictus mosquitoes were found from April to November, and the density of larval A. albopictus peaked in September (BI = 21.21), while the density of adult A. albopictus peaked in August, with a net trapping index of 2.38 mosquitoes/(net·hour). The LC50 values of delta-methrin, beta-cypermethrin, propoxur, temephos and dichlorvos were 0.017 4, 0.000 9, 0.364 1, 0.038 1 mg/L and 0.001 6 mg/L against larvae of natural strains of A. albopicchus, with RRs of 49.66, 25.53, 9.65, 2.24 and 6.06, and the mortality rates of adult mosquitoes were 66.00% (66/100), 69.39% (68/98), 25.00% (25/100), 98.97% (96/97) and 100.00% (98/98) 24 hours post-treatment with 0.08% beta-cypermethrin, 0.03% deltamethrin, 0.4% permethrin, 0.05% propoxur, and 0.5% malathion for 24 h, respectively. CONCLUSIONS: A. albopictus is widely distributed in Ningbo City, with a high population density and presents high-level resistance to common pyrethroid insecticides. The population density and insecticide resistance of A. albopictus requires to be reinforced.

Insecticide resistance of Anopheles sinensis after elimination of malaria in Henan Province, China.

BACKGROUND: Historically, malaria due to Plasmodium vivax has been epidemic in Henan Province, China, with Anopheles sinensis as the main vector. The most effective measures to prevent malaria transmission are based on vector control through the use of insecticides. However, insecticides exert a strong selective pressure on mosquito populations for insecticide resistance. The aim of this study was to investigate the susceptibility profile and population genetic characteristics of An. sinensis to provide basic data and scientific guidance for the study of resistance mechanisms and the control of An. sinensis in Henan Province. METHODS: Adult Anopheles mosquitoes were collected at sites near local farmers' sheepfolds, pigsties and/or cowsheds located in Pingqiao, Xiangfu, Xiangcheng and Tanghe counties/districts of Henan Province during July-September 2021 for insecticide susceptibility testing. Molecular identification of collected mosquitoes as belonging to genus Anopheles was by PCR, and the frequencies of mutations in the knockdown resistance gene (kdr) and acetylcholinesterase-1 gene (ace-1) were detected using gene amplification. The mitochondrial DNA cytochrome oxidase subunit I (COI) gene was amplified in deltamethrin-resistant and deltamethrin-sensitive mosquitoes to analyze the genetic evolutionary relationship. RESULTS: A total of 1409 Anopheles mosquitoes were identified by molecular identification, of which 1334 (94.68%) were An. sinensis, 28 (1.99%) were An. yatsushiroensis, 43 (3.05%) were An. anthropophagus and four (0.28%) were An. belenrae. The 24-h mortality rates of An. sinensis in Pingqiao, Tanghe, Xiangcheng and Xiangfu counties/districts exposed to deltamethrin were 85.85%, 25.38%, 29.73% and 7.66%, respectively; to beta-cyfluthrin, 36.24%, 70.91%, 34.33% and 3.28%, respectively; to propoxur, 68.39%, 80.60%, 37.62% and 9.29%, respectively; and to malathion, 97.43%, 97.67%, 99.21% and 64.23%, respectively. One mutation, G119S, was detected in the ace-1 gene. The frequencies of the main genotypes were 84.21% of specimens collected in Xiangfu (G/S), 90.63% of speciments collected in Xiangcheng (G/G) and 2.44% of speciments collected in Tanghe (S/S). Significantly higher G119S allele frequencies were observed in both propoxur- and malathion-resistant mosquitoes than in their sensitive counterparts in the Tanghe population (P < 0.05). Three mutations, L1014F (41.38%), L1014C (9.15%) and L1014W (0.12%), were detected in the kdr gene. The genotypes with the highest frequency in the populations of An. sinensis in Xiangfu and Tanghe were the mutant TTT (F/F) and wild-type TTG (L/L), at 67.86% (57/84) and 74.29% (52/70), respectively. In Pingqiao and Xiangfu, higher frequencies of the L1014F allele and lower frequencies of the L1014C allele were observed in mosquitoes resistant for beta-cyfluthrin than in those which were sensitive for this insecticide (P < 0.05). The results of Tajima's D and of Fu and Li's D and F were not significantly negative (P > 0.10), and each haplotype was interlaced and did not form two distinct branches. CONCLUSIONS: High resistance to pyrethroids and propoxur was observed at four sites, but the resistance to malathion varied according to the location. Anopheles belenrae and the L1014W (TGG) mutation in An. sinensis were first discovered in Henan Province. The deltamethrin-resistant and deltamethrin-sensitive mosquito populations showed no genetic differentiation. The generation of resistance might be the result of the combination of multiple factors.

Competitive study of homogeneous and heterogeneous Fenton-like flow-through propoxur oxidation in ROC solution.

Reverse osmosis is used as a tertiary treatment for wastewater reclamation. However, sustainable management of the concentrate (ROC) is challenging, due to the need for treatment and/or disposal. The objective of this research was to investigate the efficiency of homogeneous and heterogeneous Fenton-like oxidation processes in removing propoxur (PR), a micro-pollutant compound, from synthetic ROC solution in a submerged ceramic membrane reactor operated in a continuous mode. A freshly prepared amorphous heterogeneous catalyst was synthesized and characterized, revealing a layered porous structure of 5-16 nm nanoparticles that formed aggregates (33-49 µm) known as ferrihydrite (Fh). The membrane exhibited a rejection of >99.6% for Fh. The homogeneous catalysis (Fe3+) exhibited better catalytic activity than the Fh in terms of PR removal efficiencies. However, by increasing the H2O2 and Fh concentrations at a constant molar ratio, the PR oxidation efficiencies were equal to those catalyzed by the Fe3+. The ionic composition of the ROC solution had an inhibitory effect on the PR oxidation, whereas increased residence time improved it up to 87% at a residence time of 88 min. Overall, the study highlights the potential of heterogeneous Fenton-like processes catalyzed by Fh in a continuous mode of operation.

Bioactivity of select essential oil constituents against life stages of Anopheles arabiensis (Diptera: Culicidae).

Malaria is transmitted by infected female Anopheles mosquitoes, and An. arabiensis is a main malaria vector in arid African countries. Like other anophelines, its life cycle comprises of three aquatic stages; egg, larva, and pupa, followed by a free flying adult stage. Current vector control interventions using synthetic insecticides target these stages using adulticides or less commonly, larvicides. With escalating insecticide resistance against almost all conventional insecticides, identification of agents that simultaneously act at multiple stages of Anopheles life cycle presents a cost-effective opportunity. A further cost-effective approach would be the discovery of such insecticides from natural origin. Interestingly, essential oils present as potential sources of cost-effective and eco-friendly bioinsecticides. This study aimed to identify essential oil constituents (EOCs) with potential toxic effects against multiple stages of An. arabiensis life cycle. Five EOCs were assessed for inhibition of Anopheles egg hatching and ability to kill larvae, pupae and adult mosquitoes of An. arabiensis species. One of these EOCs, namely methyleugenol, exhibited potent Anopheles egg hatchability inhibition with an IC50 value of 0.51 ± 0.03 µM compared to propoxur (IC50: 5.13 ± 0.62 µM). Structure-activity relationship study revealed that methyleugenol and propoxur share a 1,2-dimethoxybenze moiety that may be responsible for the observed egg-hatchability inhibition. On the other hand, all five EOCs exhibited potent larvicidal activity with LC50 values less than 5 µM, with four of them; cis-nerolidol, trans-nerolidol, (-)-α-bisabolol, and farnesol, also possessing potent pupicidal effects (LC50 < 5 µM). Finally, all EOCs showed only moderate lethality against adult mosquitoes. This study reports for the first time, methyleugenol, (-)-α-bisabolol and farnesol as potent bioinsecticides against early life stages of An. arabiensis. This synchronized activity against Anopheles aquatic stages shows a prospect to integrate EOCs into existing adulticide-based vector control interventions.

Functional characterization of CYP6G4 from the house fly in propoxur metabolism and resistance.

The house fly (Musca domestica L.) (Diptera: Muscidae) is a global vector that can transmit >250 human and animal diseases. The control of house flies has heavily relied on the application of various chemical insecticides. The carbamate insecticide propoxur has been widely used for the control of house flies, and resistance to propoxur has been documented in many house fly populations worldwide. Previous studies have identified several propoxur resistance-conferring mutations in the target protein acetylcholinesterase; however, the molecular basis for metabolic resistance to propoxur remains unknown. In this study, we investigated the involvement of CYP6G4, a cytochrome P450 overexpressed in many insecticide resistant populations of Musca domestica, in propoxur metabolism and resistance by using combined approaches of recombinant protein-based insecticide metabolism and the Drosophila GAL4/UAS transgenic system. The recombinant CYP6G4 and its redox partners (NADPH-dependent cytochrome P450 reductase and cytochrome b5) were functionally expressed in Escherichia coli. Metabolism experiments showed that CYP6G4 was able to transform propoxur with a turnover rate of around 0.79 min-1. Six metabolites were putatively identified, suggesting that CYP6G4 could metabolize propoxur via hydroxylation, O-depropylation and N-demethylation. Moreover, bioassay results showed that ectopic overexpression of CYP6G4 in fruit flies significantly increased their tolerance to propoxur. Our in vivo and in vitro data convincingly demonstrate that CYP6G4 contributes to propoxur metabolism and resistance.

Composition of mosquito fauna and insecticide resistance status of Anopheles gambiae sensu lato in Itang special district, Gambella, Southwestern Ethiopia.

BACKGROUND: Anopheles arabiensis, member species of the Anopheles gambiae complex, is the primary vector of malaria and is widely distributed in Ethiopia. Anopheles funestus, Anopheles pharoensis and Anopheles nili are secondary vectors occurring with limited distribution in the country. Indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs) are pillars for the interventions against malaria control and elimination efforts in Ethiopia. However, the emergence and widespread of insecticide resistance in An. gambiae sensu lato (s.l.), might compromise the control efforts of the country. The aim of this study was to investigate composition of mosquito fauna and insecticide resistance status of An. gambiae s.l. in Itang special district ( woreda), Gambella, southwestern Ethiopia. METHODS: Adult mosquitoes were sampled from September 2020 to February 2021 using the CDC light trap and pyrethrum spray catch (PSC). CDC light traps were placed in three selected houses for two consecutive days per month to collect mosquitoes indoor and outdoor from 6:00 P.M. to 06:00 A.M. and PSC was used to collect indoor resting mosquitoes from ten selected houses once in a month from October 2020 to February 2021. Moreover, mosquito larvae were also collected from different breeding sites and reared to adults to assess susceptibility status of populations of An. gambiae s.l. in the study area. Susceptibility tests were conducted on two to three days old non blood fed female An. gambiae s.l. using insecticide impregnated papers with deltamethrin (0.05%), alpha-cypermethrin (0.05%), propoxur (0.1%), pirimiphos-methyl (0.25%) and bendiocarb (0.1%) following World Health Organization (WHO) standard susceptibility test procedure. Molecular diagnostics were done for the identification of member species of An. gambiae s.l. and detection of knockdown resistance (kdr) allele using species specific polymerase chain reaction (PCR) and allele specific PCR. RESULTS: In total, 468 adult mosquitoes were collected from different houses. Culex mosquitoes were the most dominant (80.4%) followed by Anopheles mosquitoes. Three species of Anopheles (Anopheles coustani, An. pharoensis, and An. gambiae s.l.) were identified, of which An. coustani was the dominant (8.1%) species. Higher number of mosquitoes (231) were collected outdoor by CDC light traps. Out of 468 adult mosquitoes, 294 were blood fed, 46 were half-gravid and gravid whereas the remaining 128 were unfed. WHO bioassay tests revealed that the populations of An. gambiae s.l. in the study area are resistant against alpha-cypermethrin and deltamethrin, but susceptible to bendiocarb, pirimiphos-methyl and propoxur. Of the total 86 An. gambiae s.l. specimens assayed, 79 (92%) successfully amplified and identified as An. arabiensis. West African kdr (L1014F) mutation was detected with high kdr allele frequency ranging from 67 to 88%. CONCLUSION: The detection of target site mutation, kdr L1014F allele, coupled with the phenotypic resistance against alpha-cypermethrin and deltamethrin call for continuous resistance monitoring.

[Surveillance of Aedes populations in Jiangsu Province in 2020].

OBJECTIVE: To investigate the seasonal Aedes population fluctuation and the resistance of Aedes populations to common insecticides in Jiangsu Province in 2020, so as to provide insights into vector-borne infectious diseases control. METHODS: One village was randomly sampled from each of Xinbei District of Changzhou City and Zhangjiagang County of Suzhou City in southern Jiangsu Province, Hai'an County of Nantong City and Yandu District of Yancheng City in Central Jiangsu Province, and Suining County of Xuzhou City and Sihong County of Suqian City in northern Jiangsu Province during the period between May and October, 2020. A small ponding container was sampled, and larval Aedes mosquitoes were collected using straws once each in early and late stages of each month. All larvae were bred in laboratory to adults for population identification. In addition, larval breeding were observed in all small ponding containers in and out of 30 households that were randomly sampled from six surveillance sites, and the larval mosquito density was estimated using Breteau index. Larval A. albopictus mosquitoes were sampled around Cuiyuan New Village in Jintan District of Changzhou City, and bred in laboratory to the first offspring generation, and the susceptibility of adult female mosquitoes to deltamethrin, lambda-cyhalothrin, malathion, and propoxur was tested using the filter-paper bioassay recommended by WHO. RESULTS: A total of 1 165 larval Aedes mosquitoes were captured from small ponding containers in six surveillance sites of Jiangsu Province in 2020, and all were identified as A. albopictus following eclosion. The largest number of Aedes larvae captured was found in July. A total of 1 152 households were investigated in six surveillance sites, and the mean Breteau indexes were 9.58, 13.20, 13.71, 13.20, 12.18 and 5.58 from May to October, respectively, while a high Aedes transmission risk was seen in Xinbei District of Changzhou City, with a higher Breteau index than in Suining (H = 23.667, Padjusted = 0.001) and Sihong (H = 22.500, Padjusted = 0.003) counties. The field-captured A. albopictus from Cuiyuan New Village in Jintan District of Changzhou City remained sensitive to malathion, but was resistant to propoxur, and developed high-level resistance to deltamethrin and lambda-cyhalothrin. CONCLUSIONS: A. albopictus was present in southern, central and northern Jiangsu Province in 2020, and the larval density peaked in July. A. albopictus captured from Cuiyuan New Village in Jintan District of Changzhou City has developed high-level resistance to pyrethroid pesticides.

Pulse-Exposure Toxicity of Ammonia and Propoxur to the Tropical Copepod Acartia sinjiensis.

Toxicity risk assessments of short-term discharges of contaminated waters to the aquatic environment have shown that receptor organisms can tolerate higher pulse-exposure than continuous-exposure concentrations of some contaminants. However, these observations are influenced by the mode of toxicity of the contaminants present and the concentration-time profile of the exposure. For common metal contaminants, the time-weighted average concentration (TAC) of the exposure has been useful for predicting risk of toxicity to multiple species, including the tropical, euryhaline copepod Acartia sinjiensis. To increase our understanding of the application and limitations of the TAC approach, the present study examined how varied pulse-exposure durations affect the toxicity of fast-acting contaminants, ammonia, and the common pesticide propoxur to this copepod species. Copepod larvae were exposed under continuous-exposure conditions (all life stages from eggs to nauplii to copepodites exposed) and as 6- and 18-h pulse exposures applied during the most sensitive life stage only (24-h-old nauplii) within 78-h tests. Larval development ratio and population size were assessed as test endpoints. Generally, increased exposure duration resulted in increased toxicity. Trends observed for ammonia and propoxur were slightly different for larval development and population size. Larvae tolerated greater concentrations of contaminants in a 6-h pulse (higher 10% effect concentration) than in an 18-h pulse, or a continuous 78-h exposure, whereas toxicity responses converged for the 18- and 78-h exposures. Continuous toxicity thresholds were always protective of pulse exposures, providing a conservative toxicity threshold for all durations of pulse exposures. Although generalizations for predictions of risk based on TACs are frequently effective for common metal contaminants, the TAC approach was not effective for ammonia and propoxur. Environ Toxicol Chem 2022;41:208-218. © 2021 SETAC.

Propoxur (PPx) exposure as hormonal disruptor and spermatocidal in rat model.

BACKGROUND: Propoxur is a carbamate insecticide widely used both in indoor and outdoor place to control insects. This present work was conducted to study the effect of exposure of propoxur (PPX) on hormonal and histological changes in the rat testes. METHODS: The control animals received distil water, while the treated animals received Propoxur (PPx) by inhalation every other day for one month (PPx-1) and two months (PPx-2) respectively. The animals were euthanized by cervical dislocation; blood sample was obtained for reproductive hormonal assay and the testes were excised following abdominal incision fixed in Bouin's fluid for histological observations. RESULTS: Significant decrease in the level of testosterone (TT) and increase in follicular stimulating hormone (FSH) and lutenizing hormone (LH) were observed in PPX treated groups alongside with the degenerative changes in the seminiferous tubules, complete loss of spermatogonia population, and the testicular basal membrane. There was no reversal of destruction 30 days after withdrawal of the insecticide, indicating a persistent effect. CONCLUSION: The exposure to PPX insecticide has obvious deleterious effects on rat testicular micro-structure and reproductive hormones, Therefore, inhalation of such insecticide should be limited with special care in handling to limit or minimize its hazards.

The overexpression and variant of CYP6G4 associated with propoxur resistance in the housefly, Musca domestica L.

BACKGROUND: The control of the housefly, Musca domestica, heavily relies on the application of insecticides. Propoxur, a carbamate, has been widely used for vector control. The housefly populations with high propoxur resistance display point mutations and overexpression of acetylcholinesterase. However, the roles of cytochrome P450 monoxygenases (P450s), as important detoxification enzymes, remain poorly understand in the housefly resistant to propoxur. RESULTS: P450s were implied to contribute to propoxur resistance based on the synergism of piperonyl butoxide (PBO) and the increase of P450 enzyme activity in the near-isogenic line propoxur resistant strain (N-PRS). Five P450 genes (CYP6G4, CYP6A25, CYP304A1, CYP6D3, and CYP6A1) by RNA-sequencing comparison were significantly up-regulated in the N-PRS strain with >1035-fold resistance to propoxur. A total of 13 non-synonymous mutations of three P450 genes (CYP6G4, CYP6D3, and CYP6D8) were found in the N-PRS strain. The amino acid substitutions of CYP6D3 and CYP6D8 were probably not resistance-associated single nucleotide polymorphisms (SNPs) because they were also found in the aabys susceptible strain. However, CYP6G4 variant in the N-PRS strain was not found in the aabys strain. The conjoint analysis of mutations and a series of genetic crosses exhibited that the housefly propoxur resistance was strongly associated with the mutations of CYP6G4 gene. CONCLUSION: Our results suggested that a combination of up-regulated transcript levels and mutations of CYP6G4 contributed to propoxur resistance in the housefly. © 2021 Society of Chemical Industry.

Determination of insecticides' lethal concentrations and metabolic enzyme levels in Triatoma dimidiata / Determinación de concentraciones letales y niveles de enzimas metabólicas de insecticidas en Triatoma dimidiata

Abstract: Objective: The feasibility of the use of WHO impregnated paper and biochemical assays to determine lethal concentrations (LC50 and LC99) and insecticide metabolic enzyme levels of Triatoma dimidiata. Materials and methods: LC50 and LC99 were calculated with WHO papers impregnated at different concentrations of malathion, propoxur and deltamethrin; the percentage of insensitive acetylcholinesterase (iAChE); and the levels of esterases, glutathione S-transferases, and monooxygenases in laboratory nymphs of the first stage (5 to 7 days), were undertaken using the WHO biochemical assays. Results: Respectively the LC50 and LC99 µg/cm2 obtained for malathion were 43.83 and 114.38, propoxur 4.71 and 19.29, and deltamethrin 5.80 and 40.46. A 30% of the population had an iAChE, and only a few individuals had high P450 and β-eterase levels. Conclusion: Impregnated papers and biochemical tests developed by WHO for other insects, proved to be feasible methods in monitoring insecticide resistance and metabolic enzymes involved in T. dimidiata.

Resumen: Objetivo: La factibilidad de usar los papeles impregnados y ensayos bioquímicos según la OMS para determinar concentraciones letales (CL50 y CL99) y niveles enzimáticos en la resistencia a insecticidas en Triatoma dimidiata. Material y métodos: Se calcularon la CL50 y CL99 con papeles impregnados según la OMS a diferentes concentraciones de malatión, propoxur y deltametrina; el porcentaje de acetilcolinesterasa insensible (iAChE); y los niveles de esterasas, glutatión S-transferasas, y monooxigenasas en ninfas de laboratorio del estadio I (5-7 días) se determinaron usando los ensayos bioquímicos según la OMS. Resultados: Se obtuvieron las CL50 y CL99 µg / cm2 respectivamente para malatión 43.83 y 114.38, propoxur 4.71 y 19.29, y deltametrina 5.80 y 40.46. Un 30% de las chinches tuvo iAChE, y sólo pocos individuos tuvieron niveles superiores de P450 y β-eterasas. Conclusión: Los papeles impregnados y ensayos bioquímicos que describe la OMS para otros insectos demostraron ser métodos factibles para monitorear la resistencia a insecticidas y las enzimas metabólicas involucradas en T. dimidiata.

Resistencia a insecticidas en Aedes aegypti y Aedes albopictus (Diptera: Culicidae) de Tapachula, Chiapas, México / Insecticide resistance in Aedes aegypti and Ae. albopictus (Diptera: Culicidae) populations from Tapachula, Chiapas, Mexico

Resumen: Objetivo: Determinar la resistencia a insecticidas en Ae. aegypti y Ae. albopictus de Tapachula, Chiapas, México. Material y métodos: Se utilizaron ovitrampas para obtener huevos de mosquitos Aedes y se realizaron pruebas de susceptibilidad (CDC) y ensayos enzimáticos con la primera generación. Resultados: Aedes aegypti mostró resistencia a deltametrina, permetrina, malatión, clorpirifos, temefos y a bendiocarb (CARB), mientras que Aedes albopictus a malatión y en menor grado a cloripirifos, temefos, permetrina y deltametrina. Ambas especies mostraron altos niveles de enzimas como citocomo P450 y glutatión S-tranferasa, mientras que los niveles de esterasas variaron por especie y sitio muestreado. Se detectó acetilcolinesterasa insensible a insecticidas en ambas especies. Conclusión: En un hábitat urbano de Tapachula, Chiapas, México donde se aplica control con insecticidas Ae. aegypti y Ae. albopictus sólo son susceptibles al propoxur.

Abstract: Objective: To determine the insecticide resistance status of Ae. aegypti and Ae. albopictus from Tapachula, México. Materials and methods: Mosquito eggs were collected with the use of ovitraps and CDC susceptibility bioassays and biochemical assays were conducted to determine resistance levels and resistance mechanisms, respectively. Results: Ae. aegypti showed resistance to deltamethrin and permethrin (PYRs), malathion, chlorpyrifos and temephos (OP), and to bendiocarb (CARB), while Ae. albopictus showed resistance to malathion and to a lesser intensity to chlorypirifos, temephos, permethrin and deltamethrin. Both species showed high levels of P450 and GSTs, while levels of esterases varied by species and collection site. Altered acethilcholinesterase was detected in both species. Conclusion: In an urban habitat from Tapachula, Chiapas, Mexico where vector control using insecticides takes place, Ae. aegypti and Ae. albopictus are only susceptible to propoxur.

Determination of insecticides' lethal concentrations and metabolic enzyme levels in Triatoma dimidiata.

OBJECTIVE: The feasibility of the use of WHO impregnated paper and biochemical assays to determine lethal concentrations (LC50 and LC99) and insecticide metabolic enzyme levels of Triatoma dimidiata. MATERIALS AND METHODS: LC50 and LC99 were calculated with WHO papers impregnated at different concentrations of malathion, propoxur and deltamethrin; the percentage of insensitive acetylcholinesterase (iAChE); and the levels of esterases, glutathione S-transferases, and monooxygenases in laboratory nymphs of the first stage (5 to 7 days), were undertaken using the WHO biochemical assays. RESULTS: Respectively the LC50 and LC99 µg/cm2 obtained for malathion were 43.83 and 114.38, propoxur 4.71 and 19.29, and deltamethrin 5.80 and 40.46. A 30% of the population had an iAChE, and only a few individuals had high P450 and ß-eterase levels. CONCLUSIONS: Impregnated papers and biochemical tests developed by WHO for other insects, proved to be feasible methods in monitoring insecticide resistance and metabolic enzymes involved in T. dimidiata.

OBJETIVO: La factibilidad de usar los papeles impregnados y ensayos bioquímicos según la OMS para determinar concentraciones letales (CL50 y CL99) y niveles enzimáticos en la resistencia a insecticidas en Triatoma dimidiata. MATERIAL Y MÉTODOS: Se calcularon la CL50 y CL99 con papeles impregnados según la OMS a diferentes concentraciones de malatión, propoxur y deltametrina; el porcentaje de acetilcolinesterasa insensible (iAChE); y los niveles de esterasas, glutatión S-transferasas, y monooxigenasas en ninfas de laboratorio del estadio I (5-7 días) se determinaron usando los ensayos bioquímicos según la OMS. RESULTADOS: Se obtuvieron las CL50 y CL99 µg / cm2 respectivamente para malatión 43.83 y 114.38, propoxur 4.71 y 19.29, y deltametrina 5.80 y 40.46. Un 30% de las chinches tuvo iAChE, y sólo pocos individuos tuvieron niveles superiores de P450 y ß-eterasas. CONCLUSIONES: Los papeles impregnados y ensayos bioquímicos que describe la OMS para otros insectos demostraron ser métodos factibles para monitorear la resistencia a insecticidas y las enzimas metabólicas involucradas en T. dimidiata.

[Insecticide resistance in Aedes aegypti and Ae. albopictus (Diptera: Culicidae) populations from Tapachula, Chiapas, Mexico]. / Resistencia a insecticidas en Aedes aegypti y Aedes albopictus (Diptera: Culicidae) de Tapachula, Chiapas, México.

OBJECTIVE: To determine the insecticide resistance status of Ae. aegypti and Ae. albopictus from Tapachula, México. MATERIALS AND METHODS: Mosquito eggs were collected with the use of ovitraps and CDC susceptibility bioassays and biochemical assays were conducted to determine resistance levels and resistance mechanisms, respectively. RESULTS: Ae. aegypti showed resistance to deltamethrin and permethrin (PYRs), malathion, chlorpyrifos and temephos (OP), and to bendiocarb (CARB), while Ae. albopictus showed resistance to malathion and to a lesser intensity to chlorypirifos, temephos, permethrin and deltamethrin. Both species showed high levels of P450 and GSTs, while levels of esterases varied by species and collection site. Altered acethilcholinesterase was detected in both species. CONCLUSIONS: In an urban habitat from Tapachula, Chiapas, Mexico where vector control using insecticides takes place, Ae. aegypti and Ae. albopictus are only susceptible to propoxur.

OBJETIVO: Determinar la resistencia a insecticidas en Ae. aegypti y Ae. albopictus de Tapachula, Chiapas, México. MATERIAL Y MÉTODOS: Se utilizaron ovitrampas para obtener huevos de mosquitos Aedes y se realizaron pruebas de susceptibilidad (CDC) y ensayos enzimáticos con la primera generación. RESULTADOS: Aedes aegypti mostró resistencia a deltametrina, permetrina, malatión, clorpirifos, temefos y a bendiocarb (CARB), mientras que Aedes albopictus a malatión y en menor grado a cloripirifos, temefos, permetrina y deltametrina. Ambas especies mostraron altos niveles de enzimas como citocomo P450 y glutatión S-tranferasa, mientras que los niveles de esterasas variaron por especie y sitio muestreado. Se detectó acetilcolinesterasa insensible a insecticidas en ambas especies. CONCLUSIONES: En un hábitat urbano de Tapachula, Chiapas, México donde se aplica control con insecticidas Ae. aegypti y Ae. albopictus sólo son susceptibles al propoxur.

Propoxur resistance associated with insensitivity of acetylcholinesterase (AChE) in the housefly, Musca domestica (Diptera: Muscidae).

Two unique housefly strains, PSS and N-PRS (near-isogenic line with the PSS), were used to clarify the mechanisms associated with propoxur resistance in the housefly, Musca domestica. The propoxur-selected resistant (N-PRS) strain exhibited >1035-fold resistance to propoxur and 1.70-, 12.06-, 4.28-, 57.76-, and 57.54-fold cross-resistance to beta-cypermethrin, deltamethrin, bifenthrin, phoxim, and azamethiphos, respectively, compared to the susceptible (PSS) strain. We purified acetylcholinesterase (AChE) from the N-PRS and PSS strains using a procainamide affinity column and characterized the AChE. The sensitivity of AChE to propoxur based on the bimolecular rate constant (Ki) was approximately 100-fold higher in the PSS strain compared to the N-PRS strain. The cDNA encoding Mdace from both the N-PRS strain and the PSS strain were cloned and sequenced using RT-PCR. The cDNA was 2073 nucleotides long and encoded a protein of 691 amino acids. A total of four single nucleotide polymorphisms (SNPs), I162M, V260L, G342A, and F407Y, were present in the region of the active site of AChE from the N-PRS strain. The transcription level and DNA copy number of Mdace were significantly higher in the resistant strain than in the susceptible strain. These results indicated that mutations combined with the up-regulation of Mdace might be essential in the housefly resistance to propoxur.

Analytical Methodology for Trace Determination of Propoxur and Fenitrothion Pesticide Residues by Decanoic Acid Modified Magnetic Nanoparticles.

A sensitive, rapid, reliable, and easily applicable method based on magnetic solid phase extraction (MSPE) combined with HPLC-PDA was developed for monitoring propoxur (PRO) and fenitrothion (FEN) pesticides in environmental water samples. The effect of major experimental variables on the extraction efficiency of both the pesticides was investigated and optimized systematically. For this purpose, a new magnetic material containing decanoic acid on the surface of particles was synthesized and characterized by XRD, FT-IR, SEM, EDX, and TGA analysis in detail. The simultaneous determination of pesticide molecules was carried out by using a Luna Omega C18 column, isocratic elution of acetonitrile (ACN): Water (70:30 v/v) with a flow rate of 1.2 mL min-1. After MSPE, the linear range for pesticide molecules (r2 > 0.9982) was obtained in the range of 5-800 and 10-800 ng mL-1, respectively. The limit of detections (LOD) are 1.43 and 4.71 ng mL-1 for PRO and FEN, respectively while RSDs % are below 3.5%. The applicability of the proposed method in four different environmental samples were also investigated using a standard addition-recovery procedure. Average recoveries at two spiking levels were over the range of 91.3-102.5% with RSD < 5.0% (n = 3). The obtained results show that decanoic acid grafted magnetic particles in MSPE combined with HPLC-PDA is a fast and simple method for the determination of PRO and FEN in environmental water samples.

Time-Course Changes in Urine Metabolic Profiles of Rats Following 90-Day Exposure to Propoxur.

As a major kind of carbamate insecticide, propoxur plays an important role in agriculture, veterinary medicine, and public health. The acute toxicity of propoxur is mainly neurotoxicity due to the inhibition of cholinesterase. However, little is known regarding the toxicity of propoxur upon long-term exposure at low dose. In this study, Wistar rats were orally administrated with low dose (4.25 mg/kg body weight/day) for consecutive 90 days. And the urine samples in rats treated with propoxur for 30, 60, and 90 days were collected and analyzed by employing 1H NMR-based metabolomics approach. We found that propoxur caused significant changes in the urine metabolites, including taurine, creatinine, citrate, succinate, dimethylamine, and trimethylamine-N-oxide. And the alteration of the metabolites was getting more difference compared with that of the control as the exposure time extending. The present study not only indicated that the changed metabolites could be used as biomarkers of propoxur-induced toxicity but also suggested that the time-course alteration of the urine metabolomic profiles could reflect the progressive development of the toxicity following propoxur exposure.