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Monitoring the efficacy of temephos application and use of fine mesh nylon strainers by examination of drinking water containers in guineaworm endemic villages.

Application of temephos in unsafe water sources for destroying cyclops, the intermediate hosts of guineaworm, and distribution of fine mesh nylon strainers for promoting prophylaxis against guineaworm are accepted methods of guineaworm control in different endemic countries. The existing methods of monitoring the efficacy of these guineaworm control methods are not fully informative. Examination of drinking water stored at household levels for presence of cyclops with or without Dracunculus larvae can provide information on the efficacy of these control/prophylactic methods, besides serving as a means of interpersonal health education to the community. This paper presents observations carried out in two villages in peninsular India in 1991, which revealed that while in one village complete absence of cyclops from stored water containers was attributable to the use of temephos in the village and straining of drinking water, in the other village, with no temephos application, 15.6 per cent of containers contained varying numbers of cyclops in them. Implications of these observations for guineaworm eradication activities are discussed.

Dracunculiasis↗

[Effect of sublethal concentrations of abate on biological parameters of Aedes aegypti].

The effect of abate (Temephos) applied at the rate of 0.016, 0.020 and 0.025 mg/l on fourth stadium larvae and its effect upon pupae weight, adult weight, ingested blood weight and longevity of Aedes aegypti were investigated. The insecticide was applied on F2 generation larvae that were fed an artificial diet under laboratory conditions, from larvae collected at urban level in Monterrey, N.L., Mexico. In relation to the adult, females were fed with a 10 per cent honey solution and rabbit blood for each gonotrophic cycle, while males were fed with only a honey solution. The effect of abate showed a trend to reduce the pupae weight, adult, weight and ingested blood, and to increase longevity too. Although there was not a difference for these variables in the females emerged from larvae exposed to the insecticide, the effects were significant in the males. In this sex, the weight of pupae and adult decreased by 31 and 33 per cent respectively, and longevity increased from 26 to 31 days. Possible advantages and disadvantages of these effects are discussed from a control viewpoint.

Aedes↗

Efficacy of Bacillus thuringiensis israelensis, Bacillus sphaericus and temephos for managing Anopheles larvae in Eritrea.

We evaluated the larvicidal activity of the granular formulation of Bacillus thuringiensis israelensis (Bti) serotype H-14 (Vectobac G, 200 ITU/mg) and Bacillus sphaericus (Bsph) serotype H5a5b (Vectolex CG, 670 Bs ITU/mg) against Anopheles arabiensis and other mosquitoes in breeding habitats in 3 sites, Gash-Barka, Anseba, and Debub zones, in Eritrea. The primary objective was to determine the optimal application rate and duration of effect for Bti and Bsph in representative larval habitats as compared with the organophosphate temephos. The biolarvicides were tested at 100% (high) and 50% (low) of the maximum recommended application rate. Temephos was applied at a rate of 100 ml/ha. At least 4 replicate experiments with Vectobac G (5.6 and 11.2 kg/ha), Vectolex CG (11.2 and 22.4 kg/ha) were conducted in each study site. All 3 larvicides caused significant mortality of the main malaria vector species, An. arabiensis, and other mosquito species (Anopheles cinereus, Anopheles pretoriensis, Culex quinquefasciatus). The larvicidal activity for Bti and Bsph was variable depending upon breeding habitat, mosquito species, and general ecology of the area. Both biopesticides had a similar duration of activity (2-3 wk) and were generally as effective as temephos for these time periods. In some cases, the high and low application rates for Bti and Bsph produced equivalent control over 2-3 wk. The 2 Bacillus biopesticides were less effective in habitats with high algal content and in fast flowing streams primarily because of the inability to penetrate algal mats and dilution effect, respectively. The results show that application of the 2 biolarvicides bimonthly to streambed pools, rain pools, and similar habitats would maintain control of the anopheline mosquito population.

Animals↗

Susceptibility of Italian populations of Aedes albopictus to temephos and to other insecticides.

We evaluated larval and adult susceptibility to 5 insecticides commonly used for control of Aedes albopictus, by using 15 field-collected populations from northern and central Italy, and we compared these data, collected in 2002, with those from evaluations conducted in 1992-93, 1996, and 1998-99. Larvae were tested for susceptibility to temephos and to the conventional diagnostic dosages of chlorpyrifos and fenthion proposed by the World Health Organization for Aedes aegypti. Adults were exposed to the diagnostic dosages of deltamethrin and permethrin. Overall, all of the populations tested in 2002 were still susceptible to temephos: median lethal concentration (LC50) values ranged from 0.0026 to 0.0085 mg/liter, and LC99 values ranged from 0.0093 to 0.023 mg/liter. These populations were also fully susceptible to chlorpyrifos and fenthion, and adults were susceptible to deltamethrin and permethrin. When comparing these data with those from previous evaluations, we observed a slight yet progressive increase in the LC50 and LC99 values of susceptibility to temephos, and these values are approaching the diagnostic dosage of 0.02 mg/liter proposed for Ae. aegypti. However, the implications of these findings need to be considered in light of the results of previous studies that have shown that larval Ae. albopictus are less susceptible than Ae. aegypti to organophosphate insecticides.

Aedes↗

Genotoxic evaluation of the organophosphorous pesticide temephos.

The chemical compound temephos (0,0,0',0'-tetrametyl-0,0'-thiodi-p-phenylene phosphorothioate) is an organophosphorous pesticide that has been used in Brazil since 1967 in control campaigns against the mosquito Aedes aegypti, the vector of dengue and yellow fever. We used single cell gel electrophoresis (SCGE), SOS/umu and Ames/Salmonella assays to test the toxicity and mutagenicity of temephos. Temephos was genotoxic in the SCGE assay, inducing severe DNA lesions (type IV lesions) at doses above 1.34 micro M. It was mutagenic, but not toxic, in the SOS/umu assay to Escherichia coli strain PQ37, but not to PQ35, at concentrations above 1.33 micro M, particularly when the S9 mixture was not used in the assay. Temephos was not mutagenic in the Ames assay with S. typhimurium strains TA97, TA98, TA100 and TA102, both with and without metabolic activation. However, temephos at concentrations above 3.33 micro M was mutagenic to TA98NR, YG7104 and YG7108, both with and without metabolic activation. In conclusion, temephos was genotoxic and mutagenic in all the three tests used, and in two of them at concentrations similar to those routinely used to combat Aedes aegypti.

Animals↗

Temephos resistance in two forms of Aedes aegypti and its significance for the resistance mechanism.

Aedes aegypti, at the larval stage, has been subjected to the temephos selection in laboratory. The level of temephos resistance was detected in a microplate by biochemical assay using WHO bioassay technique. The major enzyme-based resistance mechanisms involved in temephos resistance include elevated nonspecific esterase, oxidase and insensitive acetylcholinesterase. After 19 generations of temephos selection, the selected group showed resistance ratios of 4.64 and 16.92, when compared with a non-selected group and the WHO susceptible strain, respectively. The two seperated forms, type form and the pale form of Ae. aegypti showed low levels of resistance to temephos after 19 generations of selection, with resistance ratios of 4.82 and 4.07 for the type form and the pale form, respectively; when compared with the non-selected strain, 17.58 and 14.84, when compared with the WHO susceptible strain. This showed that the type form could develop higher level resistance than the pale form. The esterase inhibitor (S,S,S-tributyl phosphorotrithioate, DEF) or synergist implicated detoxifying esterase in all the temephos selected groups and the presence of elevated esterase were confirmed by biochemical assay. There were significant differences in elevated esterase activity between the temephos selected groups and the non-selected group. However no significant difference between the type form and the pale form was found. Besides the elevated esterase, there was no change in monooxygenase activity and no evidence of insensitive acetylcholinesterease for all temephos selected groups. These results suggest that temephos resistance could be developed in Ae. aegypti under selection pressure and that the main mechanism is based only on esterase detoxification.

Aedes↗

Efficacy and longevity of a new formulation of temephos larvicide tested in village-scale trials against larval Aedes aegypti in water-storage containers.

Field trials on the initial and long-term efficacy of a new formulation of temephos granules (1% on zeolite) applied at 1 ppm active ingredient (AI) were conducted in water-storage containers against Aedes aegypti in 3 villages in the Kanchanaburi Province in Thailand. A total of 316 water-storage containers of various types and sizes were included in the study. In the initial survey, we found that some containers were positive for larval Ae. aegypti, whereas others were devoid of larvae before the initiation of treatments. The containers all were numbered with paint and divided into 4 groups: with larvae and treated, without larvae and treated, with larvae untreated, and without larvae and untreated. Assessment of larval abundance was made 48 h after treatment and monthly thereafter for 5 months. Containers with larvae and that were treated exhibited almost complete absence of larval Ae. aegypti for 2 months, but a small proportion became positive after 3 months. Most of these positive containers were devoid of zeolite granules, which are visible in the containers. The number of positive containers increased in months 4 and 5, despite the fact that residues of temephos granules were present in some of the larvae-positive containers. The containers initially without larvae and treated with temephos essentially were devoid of larvae for 2 months. After 3, 4, and 5 months, about 6-23% of the containers became positive despite the fact that some had visible amounts of temephos granules. In the 2 control groups initially with and without larvae, sustained and consistent production of larvae occurred. Even in the group initially without a larval population, the containers became positive for larvae 1 month after the start of the experiment and the positivity rate increased as the trial progressed. From these studies, the conclusion can be made that a single application of temephos zeolite granules at 1 ppm AI can provide highly satisfactory control of larval Ae. aegypti in water-storage containers for at least 3 months in the field under normal water-use practices.

Aedes↗

[Aedes albopictus (Diptera: Culicidae) in Rome: experimental study of relevant control strategy parameters].

Since 1997, Aedes albopictus has colonised and then rapidly invaded the city of Rome (Italy) and its peripheral areas. Presently, the control of this mosquito in Italy relies mainly on larvicidal treatment of street storm sewer catch basins with the organophosphate temephos. We have therefore obtained baseline data on the susceptibility to temephos of the Roman Ae. albopictus population by laboratory bioassays on F1 fourth-instar larvae following standard WHO protocols. Estimated lethal concentrations were 0.014 mg/l (LC50) and 0.022 mg/l (LC90) indicating a lack of resistance to this compound. The persistence of temephos in sewer catch basins was evaluated by follow-up of catch basins treated with a dose of 1.5 mg of active ingredient. Mosquito larvae were recovered in 10% and 50% of the treated basins at 9 and 18 days posttreatment, respectively. In order to understand the relative contribution of this larval habitat to adult populations, we conducted a survey in the Zoo of Rome to estimate the larval density of mosquitoes breeding in sewer catch basins. A complete census of a 16.5 ha area mapped 243 catch basins, but only 25 (10.3%) contained water; of the latter 8 (32.0%) hosted mosquito larvae. All positive catch basins contained larvae of Culex pipiens, which were associated with Culiseta longiareolata and/or Ae. albopictus in 6 and 3 cases, respectively. A longitudinal survey of one catch basin over 4 months showed that the mean larval density of Ae. albopictus was markedly lower than that of Cx pipiens and Cs. Iongiareolata, ranging between 0 and 1.3 larvae/dip as compared to 0-33.2 and 0-22.7 larvae/dip, respectively. However, adult densities of Ae. albopictus in this area estimated during the same period with 20 ovitraps showed consistently high values (proportion of positive ovitraps around 100%). These preliminary observations suggest that whenever alternative larval biotopes other than sewer catch basins are widely available, they might be more productive and/or preferred substrates to catch basins for Ae. albopictus breeding.

Aedes↗

Larvicidal persistence of formulations of Bacillus thuringiensis var. israelensis to control larval Aedes aegypti.

After detection of resistance to the organophosphate temephos in populations of Aedes aegypti in Brazil, corncob granule (CG) and water-dispersible granule (WDG) formulations of Bacillus thuringiensis var. israelensis (Bti) were introduced in routine focal treatments. Larvicidal persistence and the influence of exposure to sunlight on VectoBac formulations of Bti were compared in 250-liter fiberglass water containers. Production of pupal Ae. aegypti in containers was used to indicate control. In untreated containers, survival of larvae was always above 95%. A temephos sand granule formulation used as reference treatment maintained 100% control throughout the 12-wk period in all situations. Under sunlight exposure, control dropped below the 90% level in the 2nd week after treatment at both dosages of VectoBac CG (1 and 2 g/50 liters) and VectoBac tablet (T) formulation at 1 tablet/100 liters. VectoBac T at 1 tablet/50 liters provided 2 wk of 100% control. VectoBac WDG at dosages of 1 and 2 g/500 liters provided 100% control for 3 wk. Without sunlight exposure (covered containers), VectoBac CG provided 9 wk of continuous 100% control and 5 wk of continuous 100% control, respectively, at 1 and 2 g/50 liters. The VectoBac T formulation at both dosages initially provided 2 wk of 100% control. After this period, the control level fluctuated between 96 and 100%. VectoBac WDG provided continuous 100% control for 7 wk for the lower dosage and for 6 wk for the higher dosage. At both dosages of WDG, 100% control was achieved in 11 wk out of the 12-wk period.

Aedes↗

Comparison of three larviciding options for malaria vector control.

A field study was carried out over 27 weeks in the south Batinah region of Oman to assess the efficiency and cost-effectiveness of different strategies for vector control of malaria. Three larviciding strategies for Anopheles spp. were applied to intervention areas and compared with a control area, with over 2000 breeding sites monitored for 6 months. The normal method of spraying 1 ppm temephos larvicide fortnightly was found to be less efficient and less cost-effective than using 0.5 ppm temephos applied weekly. A third, more environmentally favourable method, to search for vector larvae and treat only those breeding places, was more effective than fortnightly spraying but less effective than the weekly half dose and was the most expensive strategy.

Animals↗

Operational use of neem oil as an alternative anopheline larvicide. Part A: Laboratory and field efficacy.

We conducted a study to determine the laboratory and field efficacy of neem oil towards anopheline larvae. No difference in LC50 was observed between laboratory and field strains for temephos, chlorpyriphos-methyl/fenitrothion and neem oil. No difference in susceptibility was found after 3 months of application every 2 weeks. Water treated with a single application of traditional larvicides was free of larvae after 4 weeks; neem oil-treated water, however, was free after 2 weeks but not at 4 weeks. Application of chlorpyriphos-methyl/fenitrothion and neem oil every 2 weeks for 7 rounds resulted in dramatic reduction in larval density with no statistically significant differences. An adult survey after larviciding also showed no significant difference. The efficacy of crude neem oil appears to be below that of conventional larvicides.

Animals↗

Operational use of neem oil as an alternative anopheline larvicide. Part B: Environmental impact and toxicological potential.

This study was conducted to investigate the preliminary environmental and mammalian toxicology of neem oil, temephos and chlorpyriphos-methyl/fenitrothion. Culex pipiens, Daphnia magna and Gambusia affinis were used to study environmental impact. A high level of toxicity was observed, with slight differences between organisms. The emulsifiers individually also displayed toxicity towards the tested organisms. Up to 90 days daily oral crude neem oil treatment (5 g/kg body weight) of laboratory mice did not cause any significant changes in weekly body weight gain, nor in serum liver damage indicators, direct bilirubin or total bilirubin. Blood parameters of treated mice up to 90 days were not statistically different from those of control mice. Neem oil could be used as an environmentally friendly alternative to the traditional chemical anopheline larvicides.

Alanine Transaminase↗

[Resistance to insecticides in larvae and adults of Aedes aegypti, Havana City: prevalence of A4 esterasa associated with resistance to temephos].

2 strains of Aedes aegypti from 2 people's councils with high indexes of infestation of this vector were studied due to the need of carrying out an effective control of larvae and adults in the municipality of Guanabacoa. The levels of susceptibility and/or resistance to organophosphate insecticides, pyrethroids and a carbamate were determined. The results of the bioassays in larvae showed a complete susceptibility to organophosphate insecticides, malathion, clorpirifos, methyl-pyrimifos and propoxur carbamate in both people's councils. However, it was observed a high resistance to temephos and fenthion. No resistance to fenitrothion was found in one of the people's councils. At the adult stage and at the dose recommended by the World Health Organizaton or by the product manufacturers, it was observed resistance to malathion, fenitrothion and propoxur; nevertheless, better results were obtained with the pyrethroids, with mortality percentages over 90%. According to the results obtained, by using the SSS tributyl phosphotritiade sinergist (DEF) and piperonyl butoxide, it was demonstrated that the multiple function esterases and oxidases played an important role in the resistance to temephos and fenthion. It was proved that he amplified activity of these enzymes was at an elevated frequency in both people's councils, the same as the glutathione transferase (GST) in one of the 2 people's councils. It was observed that in both people's councils prevailed the amplified activity of A4 esterases by polyacrylamide gel electrophoresis.

Aedes↗

Bottle and biochemical assays on temephos resistance in Aedes aegypti in Thailand.

The bottle bioassay measuring the time-mortality rate is a simplified procedure for detecting insecticide resistance. It can be used with a biochemical microplate assay to identify the mechanism involved. This integrated approach was used to detect temephos resistance in Aedes aegypti from Nonthaburi (lowest use) and Roi Et (highest use). Ae. aegypti BKK1 laboratory strain was used as the susceptible reference strain. The appropriate concentration of insecticide for bottle bioassay was determined empirically for Ae. aegypti BKK1 strain and found to be in the range of 800-1,050 microg/bottle. The time-mortality rate at 800 microg/bottle was 170 +/- 8.66 minutes, significantly different from the time-mortality rates in the 850, 900, 950, and 1,050 microg/bottle (p = 0.008) concentrations, which were 135 +/- 15.00, 140 +/- 8.66, 135 +/- 15.00, and 125 +/- 8.66 minutes, respectively. The cut-off concentration selected for resistance detection was 850 microg/bottle. The time-mortality rate for the Roi Et strain was 382 +/- 26.41 minutes, significantly higher than the Nonthaburi (150 +/- 25.10 minutes) and BKK1 strains (145 +/- 20.49 minutes) (p < 0.001). The temephos resistance ratio (RR100) for the Ae. aegypti Roi Et strain was 2.64-fold higher at lethal time (LT100) than for the reference Ae. aegypti BKK1 strain. The mean optical density (OD) value from the biochemical microplate assay for the non-specific esterase of the Roi Et strain was higher than the mean OD for the non-specific esterase of both the Nonthaburi and BKK1 strains. Insensitive acetylcholinesterase was not found to be responsible for the resistance in the field-collected mosquitos. This study suggests that esterase detoxification is the primary cause of resistance in the Ae. aegypti population from Roi Et. Both the bottle bioassay and the biochemical microplate assay were proven to be promising tools for initial detection and field surveillance for temephos resistance.

Acetylcholinesterase↗

Sequential release and residual activity of temephos applied as sand granules to water-storage jars for the control of Aedes aegypti larvae (Diptera: Culicidae).

Two long-term experiments were carried out on the release profile and efficacy of temephos 1% GR (sand granules) against Aedes aegypti larvae in water-storage containers. In the first experiment, the efficacy of temephos 1% GR enclosed and tied in a muslin cloth and placed in water at the bottom of 200 L earthen water-storage jars was studied by exposing the packets for four to nine wk in one set ofjars and then transferring them sequentially to new sets ofjars four times successively. Temephos released slowly from the granules, the magnitude of release being adequate in the initial period of two to three wk after treatment. Following this period, the efficacy of the granules increased substantially where 92-100% inhibition of emergence even at the lowest dosage of 1 g/100 L (0.05 mg/L AI) was obtained for about another five mo or longer. On removal of the packets from a given set of jars, the released residues remaining in the jars and water lasted a maximum of one to six wk post-removal depending on the magnitude of prior release into the jars. This experiment provided clear evidence that temephos is released slowly over a long period of time in water-storage jars. In the second experiment, we compared the efficacy of temephos 1% GR at 1 and 10 g (0.05 and 0.5 mg/L AI) per 200-L water in jars painted and unpainted on the inside. The efficacy in the painted jars, although high, was consistently lower than that in the unpainted jars, where 99-100% control of larvae was achieved at both rates for a minimum of five mo after treatment. On the basis of this experimental evidence, it is desirable to study the efficacy of lower dosages of temephos than those currently used in Ae. aegypti control programs. The use of controlled release formulations or sachets that are retrievable during cleaning and washing will be more practical and desirable. Both of these interventions will make the program more cost effective.

Aedes↗

Evaluation of various control agents against mosquito larvae in rice paddies in Taiwan.

A field test was conducted in rice paddies adjacent to Wufeng, Taichung County in Central Taiwan to evaluate the efficacy of control agents against mosquito larvae. The agents included Bacillus thuringienesis israelensis (Bti), two Lagenidium giganteum products (Lg product A and T), and temephos. The major mosquito species found in the rice paddies were Culex tritaeniorhynchus/vishnui and Anopheles sinensis. Compared to controls, a 7-day treatment with Bti or Lg products A and T caused overall reductions in the number of immatures (larvae and pupae) of Cx. tritaeniorhynchus/vishnui of 77.5%, 49.7%, and 21.9%, respectively, whereas temephos caused an increase of 66.9%. The overall reductions in An. sinensis were 85.4%, 8.6%, 44.6%, and 92.1%, respectively. There was no significant reduction in the number of mosquito larvae following 42 days of treatment with these agents. In summary, 1-week treatments with both biological control agents produced moderate overall reductions in mosquito larvae in rice paddies. The insecticide temephos, on the other hand, was very effective at suppressing the larvae of An. sinensis but significantly increased the number of Cx. tritaeniorhynchus/vishnui larvae in temephos-treated plots.

Animals↗