Impact of environmental temperatures on resistance to organophosphate insecticides in Aedes aegypti from Trinidad / Repercusión de las temperaturas ambientales sobre la resistencia de Aedes aegypti a los insecticidas organofosforados en Trinidad

OBJECTIVE: To examine the effects of increasing larval rearing temperatures on the resistance status of Trinidadian populations of Aedes aegypti to organophosphate (OP) insecticides. METHODS: In 2007-2008, bioassays and biochemical assays were conducted on A. aegypti larvae collected in 2006 from eight geographically distinct areas in Trinidad (Trinidad and Tobago). Larval populations were reared at four temperatures (28 ± 2ºC, 32ºC, 34ºC, and 36ºC) prior to bioassays with OP insecticides (fenthion, malathion, and temephos) and biochemical assays for esterase enzymes. RESULTS: Most larval populations reared at 28 ± 2ºC were susceptible to fenthion (>98% mortality) but resistant to malathion and temephos (< 80% mortality). A positive association was found between resistance to OP insecticides and increased activities of α- and β-esterases in larval populations reared at 28 ± 2ºC. Although larval populations reared at higher temperatures showed variations in resistance to OPs, there was a general increase in susceptibility. However, increases or decreases in activity levels of enzymes did not always correspond with an increase or decrease in the proportion of resistant individuals reared at higher temperatures. CONCLUSIONS: Although global warming may cause an increase in dengue transmission, based on the current results, the use of insecticides for dengue prevention and control may yet be effective if temperatures increase as projected.

OBJETIVO: Examinar los efectos del aumento de las temperaturas de desarrollo larvario sobre el estado de resistencia a los insecticidas organofosforados de las poblaciones de Aedes aegypti en Trinidad. MÉTODOS: En 2007 y 2008 se llevaron a cabo ensayos biológicos y bioquímicos en larvas de A. aegypti recogidas en el 2006 de ocho áreas geográficamente separadas en Trinidad (Trinidad y Tabago). Las poblaciones larvarias se desarrollaron en cuatro temperaturas (28 ± 2 ºC, 32 ºC, 34 ºC y 36 ºC) antes de los ensayos biológicos con insecticidas organofosforados (fentión, malatión y temefós) y los análisis bioquímicos para las enzimas de esterasa. RESULTADOS: La mayoría de las poblaciones larvarias que se desarrollaron a 28 ± 2 ºC fueron susceptibles al fentión (mortalidad > 98%) pero resistentes al malatión y al temefós (mortalidad < 80%). Se encontró una asociación positiva entre la resistencia a los insecticidas organofosforados y la mayor actividad de αy β-esterasas en las poblaciones larvarias que se desarrollaron a 28 ± 2 ºC. Aunque las poblaciones larvarias que se desarrollaron a temperaturas mayores mostraron variaciones en la resistencia a los organofosforados, hubo un aumento general de la sensibilidad. Sin embargo, los aumentos o las disminuciones en los niveles de actividad de las enzimas no siempre se correspondieron con un aumento o disminución en la proporción de individuos resistentes desarrollados a las temperaturas más altas. CONCLUSIONES: Aunque el recalentamiento del planeta puede causar un aumento de la transmisión del dengue, según los resultados de este estudio el uso de insecticidas para la prevención y el control del dengue todavía puede ser eficaz si las temperaturas aumentan según lo proyectado.

Susceptibility status of Japanese encephalitis vectors in Kurnool and Mehboobnagar districts of Andhra Pradesh, India.

An entomological survey in Kurnool and Mahboobnagar districts of Andhra Pradesh state, India was undertaken in January, 2002 to find out vector population of Japanese Encephalitis and their susceptibility status to different insecticides. Five Culicine species viz. of Cx. tritaeniorhynchus, Cx. vishnui, Cx. pseudovishnui, Cx. bitaeniorhynchus, Cx. gelidus were captured during the survey. Another suspected vectors Anopheles subpictus and An. hyrcanus were also prevalent in the study villages. The larval population showed a high degree of resistance to Fenthion and Temephos. The test mortality to DDT ranged between 75% and 90%. The mortality of Cx. tritaeniorhynchus, Cx vishnui and An. subpictus ranged from 85% to 95%. Cx. tritaeniorhynchus is susceptible to Deltamethrin and Cyfluthrin. Cx. vishnui showed 80 % and 85% mortality to DDT and Malathion respectively.

Mice testicular damage elicited by malathion

Organophosphorous pesticides are a healh problem worldwide, mostly for agroworkers, which are around 2600 millions of person in the year 2001. Little is known about male reproductive damage elicited by these chemicals. The acute effects of malathion (single intraperitoneal injection), 1/12 the LD50 to mice were analyzed at 1, 8, 16, 35 and 40 days after injection, in terms of spermatogenic cell proliferation and apoptosis and of Sertoli cell compromise as revealed by immunocytochemical detection of CK-18 (cytokeratin). The micronuclei test was done to assess for genotoxic activity of the agropesticide. The results showed decreased germ cell number followed by compensatory spermatogonial proliferation by 16 days (as shown by thymidine-H3 testicular uptake), increased apoptotic rate, mainly of spermatogonia, and preleptotene spermatocytes. Sertoli cell reactivity to CK-18, denoting alteration of them, probably due to germ cell damage, was seen in treated mice. Genotoxicity for somatic cells was demonstrated by the micronuclei test. Therefore, it is assumed that Sertoli as well as Leydig cells are affected together with spermatogenesis; recuperation of testicular morphology is seen by longer time intervals.

Insecticide cross-resistance spectra and underlying resistance mechanisms of Sri Lankan anopheline vectors of malaria.

Present status of insecticide resistance was investigated in two major vectors of malaria; An. culicifacies and An. subpictus, collected from a high malaria transmission area in Sri Lanka during 1996/1998. Adult and larval bioassays were carried out to obtain log-probit mortality lines for malathion, propoxur, permethrin and chlorpyrifos. Respective LD50 values were 4.45%, 0.002%, 0.16% and 0.001% for An. culicifacies and 0.66%, 0.004%, 0.004% and 0.04% for An. subpictus. Adults were also tested for WHO standard discriminating dosages of malathion, propoxur, permethrin, DDT, cypermethrin, deltamethlin and lambda cyhalothrin. Both populations were highly resistant to DDT. An. culicifacies was more resistant to malathion and An. subpictus was more resistant to chlorpyrifos. About 25% of both populations were resistant to permethrin. An. culicifacies was susceptible to propoxur, deltamethrin and lambda cyhalothrin and An. subpictus to cypermethrin and lambda cyhalothrin. Adult mosquitos were individually tested for their insecticide detoxifying enzyme activities and altered target-site, acetylcholinesterase. High general esterase activity indicated the presence of amplified esterase genes in both populations. Native gel electrophoresis resolved one elevated esterase isoenzyme, with high affinity to organophosphates, from each species. Malathion carboxylesterase mechanism was present in both populations. Higher glutathione-S-transferase activity was marked in An. subpictus. Synergistic studies showed the possible involvement of monooxygenases in resistance in both species. Acetylcholinesterase activity of approximately 80% of both populations was not inhibited by a standard dosage of propoxur. Low resistance to carbamates shows that the impact of agricultural pesticides is not significant in the development of resistance especially in An. culicifaies. Pyrethroids, other than permethrin, can be successfully used in vector control programs. Carbamates will be an alternative.

Rate of resistance development in wild Culex quinquefasciatus (Say) selected by malathion and permethrin.

Wild caught female Culex quinquefasciatus (Say) from Kuala Lumpur were blood fed and reared in the insectarium. The late third stage of the F1 larvae which survived the high selection pressure of malathion and permethrin were reared and colonies were established from adults that emerged. Larvae from these colonies were then subjected in the subsequent 9 generations to higher selection pressure. The rate of resistance development were measured by LC50 value of larval bioassay, LT50 value of adult bioassay and the frequency of the elevated esterase levels. In another set of experiments using the same batch of Culex mosquitos, the larvae were not exposed to any insecticides and the decrease in resistance rate was monitored in each subsequent 9 generations by using similar methods. The heterozygous standard laboratory strain was selected for susceptibility using the single raft sib-selection method. The result showed that the field collected F1 generation was 96.0 and 6.3 fold more resistant to malathion and permethrin, respectively. After selection for about 9 generations the resistance ratio to malathion and permethrin was 6.2 and 767.3 fold more compared to the LC50 values of F1 generations, respectively. Esterase in F1 larvae was 6.0 fold more than the standard laboratory strain.

Interaction of organophosphorus insecticides phosphamidon & malathion on lipid profile & acetylcholinesterase activity in human erythrocyte membrane.

The organophosphorus insecticides phosphamidon and malathion were found to inhibit the activity of human acetylcholinesterase in vitro, in the human erythrocyte membrane. Lineweaver-Burk analysis indicated that the insecticide induced inhibition of acetylcholinesterase activity was uncompetitive in nature. The total lipid, cholesterol and phospholipid contents of erythrocyte membranes were reduced following phosphamidon and technical malathion treatment, while the level of lipid peroxidation was raised following malathion treatment.

Insecticide susceptibility status of Anopheles koliensis (Diptera: Culicidae) in northeastern Irian Jaya, Indonesia.

Anopheles koliensis, an important malaria vector in the interior region of Irian Jaya, Indonesia, was evaluated for susceptibility to three different insecticide compounds using the standard World Health Organization diagnostic test kit and pretreated impregnated papers. A series of tests were conducted in Arso PIR I, a transmigrant settlement 60 km south of Jayapura, from January 1988 to May 1989. All compounds were tested at the recommended diagnostic dosage and exposure time. An. koliensis were susceptible to 1.0% fenitrothion at two hour exposure (N = 358) and 5.0% malathion at one hour exposure (N = 371) after the 24-hour holding period. Significant resistance to DDT was observed in both the An. koliensis and Culex quinquefasciatus populations. Approximately 30% of the An. koliensis population (N = 468) was resistant to 4% DDT at both one and two hour exposures. These findings indicate that routine use of DDT in Arso PIR I for indoor residual house spraying may be of limited effectiveness, in part, because of physiological resistance. However, use of an alternative insecticide will be more expensive and might prove equally ineffective because of the exophilic behavior of the species. This is the first confirmed report from repeated observations of DDT resistance in An. koliensis from Indonesia.

Residual effect of four insecticides applied for indoor control of phlebotomies papatasi [scopoli]

The residual effect of four known insecticides prepared in olive oil and sprayed on cement plastered walls was estimated against the Egyptian strain of Phlebotomus papatasi [Scopoli]. The results showed that propoxur reduced the number of the tested sand flies to 16.5%, 24.1%, 46.4%, 53.4% and 76.7%, 75 days post treatment after an exposure time for 5, 10, 15, 20 and 30 minutes. Permethrin caused zero, 17.5%, 25.0%, 41.4%, and 51.9%, malathion caused 4.3%, 10%, 26.7%, 35.9%, and 49.1%; and BHC caused zero, 10%, 30%, 36.3% and 48.1% for the same exposure time, respectively. This indicated that propoxur had the highest residual effect and BHC had the lowest one. The results were discussed in relation to leishmaniasis control

Therapeutic action of acetylcholine in malathion toxicity.

Administration of malathion to the developing Philosamia ricini larvae induces accumulation of acetylcholine, marked inhibition of acetylcholinesterase activity, depletion of all nutrients, heavy weight loss and high mortality. Pre-treatment of the larvae with acetylcholine via feed, reduces malathion toxicity and conversely, feeding of acetylcholine to malathion-treated larvae reverses the toxic effects. Resumption of normal control feed to malathion-treated insects results in higher mortality than in insects fed acetylcholine after exposure to malathion. This emphasizes the therapeutic action of acetylcholine. Feeding of a mixture of equal quantities of malathion and acetylcholine recorded significantly lower mortality among insects in comparison to those fed malathion alone. This further supports the protective action of acetylcholine. Reversal of malathion toxicity and the protective action of acetylcholine have been attributed to the mediation of choline, an essential insect vitamin that gets released as a catabolic product of acetylcholine.

Insecticide susceptibility studies of three cryptic species of the Anopheles balabacensis complex.

Susceptibilities of two colonies of the taxon An. dirus (one from Perlis and from Thailand) and one colony of An. balabacensis from Sabah to DDT, dieldrin, malathion, fenitrothion and propoxur were determined. DDT and fenitrothion tolerance was found in An. dirus species B and An. balabacensis. No resistant strain was isolated as the two colonies were not either homozygous or nearly so for resistance. Field testing of the susceptibility of the adults of An. balabacensis to DDT was carried out between 1957 to 1976. The results indicated a progressive rise in the LC50 levels greater than 1% in almost all instance. The variation in the number of sprays applied in some districts have resulted in varying sensitivities. Association between the changes in levels of DDT susceptibility and exophilic habit of An. balabacensis has been observed but needs further confirmation. The significance of these findings and the difficulties in distinguishing tolerant from truly resistant individuals are discussed in relation to accurate species identification.