To determine resistance level and characterize malathion and permethrin resistance in Culex quinquefasciatus, two methods were used namely: WHO procedures of larval bioassay to determine the susceptibility of lethal concentration (LC) and adult bioassay to determine the lethal time (LT) which are resistant to malathion and permethrin. These mosquito strains were bred in the Insectarium, Division of Medical Entomology, IMR. Thousands of late fourth instar larvae which survived the selection pressure to yield 50% mortality of malathion and permethrin were reared and colonies were established from adults that emerged. Larvae from these colonies were then subjected to the subsequent 10 generations in the test undertaken for malathion resistant strain (F61 - F70) and permethrin resistant strain (F54 - F63). Selection pressure at 50% - 70% mortality level was applied to the larvae of each successive generation. The rate of resistance development and resistance ratio (RR) were calculated by LC5 0 for larval bioassay and LT50 value for adult bioassay. The lab bred Cx. quinquefasciatus was used as a susceptible strain for comparison purpose. The adult bioassay test was carried out by using diagnostic dosages of malathion 5.0%, permethrin 0.75% and with propoxur 0.1%. All bioassay results were subjected to probit analysis. The results showed that LC5 0 for both malathion (F61 - F70) and permethrin (F54 - F63) resistant Cx. quinquefasciatus increased steadily to the subsequent 10 generations indicating a marked development of resistance. The adult female malathion resistant strain have developed high resistance level to malathion diagnostic dosage with resistance ratio 9.3 to 9.6 folds of resistance. Permethrin resistance ratio remained as 1.0 folds of resistance at every generation. It was obvious that malathion resistance developing at a higher rate in adult females compared to permethrin. Female adults exposed to 2 hours of exposure period for propoxur 0.1% showed presence of cross-resistance among the both strains of mosquitoes towards propoxur and it was indicated by 70%-100% mortality at 24 hours post-recovery period.
The susceptibility of Culex quinquefasciatus to chemical insecticides in two field sites in Kuala Lumpur was evaluated using the WHO standard susceptibility test. Less then 7 days old female mosquitos, reared from wild caught females were exposed to discriminating dosages of insecticides at recommended exposure periods. The larval bioassay were conducted using the multiple concentrations and the LC50 value was determined. The results indicated that cyfluthrin is the most effective among all the insecticides tested with LT50 value of 29.95 min and 28.59 min, for the strain from Ampang Hill and Pantai Dalam, respectively. It was surprisingly to note that both these field strains showed 0% mortality when tested against malathion and DDT. The LC50 value indicated that both strains were highly resistant to malathion with resistance ratio of 17,988 folds and 14,053 folds, respectively. This concludes that resistance at larval stages is extremely high compared to adult stages.
Laboratory-bred females of Culex quinquefasciatus, Aedes aegypti and Aedes albopictus from the insectarium, Unit of Medical Entomology, Institute for Medical Research were used in the experiment. The late third stage of the F0 larvae which survived the high selection pressure of malathion, permethrin and temephos were reared and colonies were established from adults that emerged. Cx. quinquefasciatus larvae were subjected to selection by malathion and permethrin for 40 generations, Ae. aegypti larvae to malathion, permethrin and temephos for 32 generations and Ae. albopictus larvae were selected against malathion and permethrin for 32 generations and 20 generations against temephos. The rate of resistance development was measured by LC50 value. Cx. quinquefasciatus larvae developed higher resistance to malathion and permethrin compared to Ae. aegypti and Ae. albopictus. On the whole, permethrin resistance developed at a faster rate than malathion and temephos.
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.
Cyfluthrin (Solfac ULO15) and malathion 96 TG were evaluated against sentinel sugar-fed adults and 4th-instar larvae of Aedes aegypti at high-rise flats in Malaysia by ULV spraying. The impact of both insecticides on field populations of Aedes spp. (Ae. aegypti and Ae. albopictus) larvae were monitored weekly using containers. Both cyfluthrin and malathion 96 TG showed adulticidal effects but cyfluthrin showed more significant larvicidal effect than malathion 96 TG (P < 0.05).
The insecticide resistance status of 4 strains of adult male Blattella germanica, viz M (Malacca), E (England), F (restaurant) and K (cafeteria) against malathion and bendiocarb compared with a reference susceptible strain (S) was determined by using a modified WHO bioassay method. The results indicated that all the 4 strains were resistant to the insecticides albeit in different degrees. Resistance ratios for malathion ranged from 1.85-41.07-fold, whereas that of bendiocarb ranged from 1.68-4.83-fold. The biochemical microplate enzyme assays technique employed indicated that the resistance in M and E strains were attributed to acetylcholinesterase insensitivity. Multiple resistance was not detected in any of the 4 strains. Parameters of the identified resistance mechanism correlated well with the observed level of resistance. Agar gel electrophoresis showed that variations in esterase isoenzymes did not confer organophosphate and carbamate resistance to the 4 strains.
In an effort to develop a more effective technique in dispersing a microbial control agent, Bacillus thuringiensis (Bt), a truck-mounted ultra low volume (ULV) generator (Scorpion) was used to disperse B. thuringiensis israelensis (Bti) and Bti with malathion. Complete larval and adult mortalities for all tested mosquito species within the first 70-80 feet from the ULV generator were achieved. Beyond that distance less than 50% mortality was achieved as insufficient sprayed particles reached the area. A minimum of 10(3) Bti colony forming units per ml is required to cause 100% larval mortality. The sprayed Bti larvicidal toxins were persistent in the test water 7 days post ULV. The effectiveness of B. thuringiensis jegathesan (Btj), a new mosquitocidal Bt serotype was also evaluated. Similar mortality results as Bti were achieved except that the Btj toxins underwent degradation in the test water, since less than 50% less in larval mortality was observed in 7 days post ULV samples. This ULV method has the potential to disperse Bt and malathion effectively for a simultaneous control of mosquito adults and larvae.
The ability to identify the occurrence of different resistance genotypes in field populations of mosquito is considered important for the purpose of optimising chemical control operations. The recent development of rapid microassays of enzymes responsible for resistance has provided a means for rapidly assessing the genetic background of target mosquito populations. This concept is the topic of investigation in this study. Non-specific esterase activity, which is responsible for the resistance to organophosphates in Malaysian Culex quinquefasciatus Say adults, was determined in 3 field populations from Kuala Lumpur City using rapid enzyme assay. The optical density results were used to estimate the genotypic frequencies of the populations. Subsequently, time-dependent changes in the various frequencies were determined. Such techniques allowed rapid assessment of resistance genotypes for decision-making and its possible use in insect control merits further investigation.
Two field trials in the control of subperiodic brugian filariasis vectors, mainly Mansonia bonneae and Mansonia dives were carried out in Sarawak, East Malaysia. In the first trial, malathion ultra-low volume (ULV) spray was used to control the Mansonia mosquitos in two filariasis endemic villages. Six spray rounds were applied at biweekly intervals at Kampung Rasau and two spray rounds were applied at monthly intervals in Kampung Triboh. ULV malathion spray reduced biting Ma. bonneae population for 3 days after spraying. The biting density decreased to 50% of the pre-treatment level by the 12th - 13th day and reached the pre-treatment level by the 24th - 25th day. Contact bioassay tests on caged Mansonia mosquitos revealed considerable penetration of the malathion aerosol indoors and relatively adequate coverage outdoors. The estimated number of bites per case per day was 1.09 to 4 times less in the sprayed kampung than in an unsprayed control kampung. The parous and daily survival rates of Mansonia mosquitos were not significantly affected by the spraying. In a second trial, chemotherapy with diethylcarbamazine citrate (DEC) was combined with vector control through indoor residual spraying in Kampung Ampungan. The results were compared with the use of only DEC mass treatment in Kampung Sebangkoi and Kampung Sebamban. The combined control measures in Kampung Ampungan reduced the MfD-50 to 44% of the pre-treatment level over a period of 4 years. In the other two kampungs where only mass DEC therapy was applied, the microfilarial rate and MfD-50 declined significantly in the second blood survey but increased gradually in two subsequent follow-up blood surveys. The total insecticidal impact for Ma. bonneae was 3.9 to 1 indoors and 2.7 to 1 outdoors. These results indicated that quarterly pirimiphos-methyl indoor spraying used in integrated control could reduce indoor transmission by 3.9 times. The infective rate from the Ma. bonneae dissected in all three kampungs after the interventions, irrespective of DEC treatment alone or in combination with pirimiphos-methyl residual spraying were reduced by two fold. However the infection rate of brugian filarial larvae in Kampung Ampungan was significantly reduced after the use of DEC and insecticide. Annual Transmission Potential (ATP) showed a high significant reduction in Kampung Ampungan (p > 0.001) compared with Kampungs Sebangkoi and Schambam. In Ampungan, the ATP was reduced by 8.5 times indoors after the MDA and insecticidal application and 3 times outdoors. The reduction rate for Sebangkoi and Sebamban both indoors and outdoors were less than 2 fold.(ABSTRACT TRUNCATED AT 400 WORDS)
Rapid enzyme microassays for the detection of resistance due to organophosphate and carbamate in individual field-collected strains of Culex quinquefasciatus adults were conducted. These tests allowed accurate differentiation by eye, on the basis of color changes of susceptible and resistant individuals. Two separate tests were conducted for the biochemical assays. In the insensitive acetylcholinesterase (AChE) test, acetylthiocholine iodide (ACTH) and 5,5-dithiobis-(2-nitrobenzoic acid) (DTNB) were used as substrate and coupling agent respectively. The resulting yellow chromophore indicated AChE activity. Test results showed that the color intensity decreased as increasing concentrations of propoxur were added, thereby confirming the susceptibility of the enzyme to inhibitor. Assay of non-specific esterase however, indicated elevated levels which were correlated with degree of malathion resistance. Electrophoretic data revealed the presence of 2 esterase bands in all strains. It was concluded that such a pattern was not contributory to malathion resistance in adults.
Paddy (unmilled rice), milled rice and maize-bound 14C residues were prepared using 14C-succinate-labelled malathion at 10 and 152 ppm. After 3 months, the bound residues accounted for 12%, 6.5% and 17.7% of the applied dose in paddy, milled rice and maize respectively in the grains treated at 10 ppm. The corresponding values for the 152 ppm were 16.6%, 8.5% and 18.8%. Rats fed milled rice - bound 14C-residues eliminated 61% of the 14C in the faeces and 28% in the urine. The corresponding percentages for paddy and maize were 72%, 9% and 53%, 41% respectively; indicating that bound residues from milled rice and maize were moderately bioavailable. When rice-bound malathion residues (0.65 ppm in feed) were administered to rats in a 5 week feeding study, no signs of toxicity were observed. Plasma and RBC cholinesterase activities were slightly inhibited: blood urea nitrogen was significantly elevated in the test animals. Other parameters examined showed no or marginal changes.
Five pesticides were evaluated against laboratory colonies of Leptotrombidium fletcheri (Womersly and Heaslip) by the Pasteur pipet technique. The pesticides were dieldrin (LC50 = 3.6 ppm, LC99 = 18.2 ppm), bromopropylate (LC50 = 9.2 ppm, LC99 = 239.6 ppm), dicofol (LC50 = 27.8 ppm, LC99 = 118.1 ppm), fenthion (LC50 = 15.4 ppm, LC99 = 29.7 ppm), and malathion (LC50 = 84.7 ppm, LC99 = 313.9 ppm). Dieldrin was the most toxic. Dicofol was recommended for further evaluation in field trials.
Field trials were conducted in two residential areas of Petaling Jaya Municipality to test the adulticidal and larvicidal effects of malathion 96% TG and Resigen on Aedes aegypti. Malathion is the currently used insecticide in Malaysia for the control of dengue. The Leco HD ULV machine was used throught the trials. For malathion the flow rate was 90 ml/minute at a vehicle speed of 8kph and for Resigen the flow rate was 200 ml/minute at the same vechicle speed. Malathion was more effective giving higher mortality rates when compared with Resigen. The mortality rate of adult Ae. aegypti outdoor was higher than in the living room and kitchen. Both insecticides did not show promising larvicidal effects.
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.
A survey of 4.112 primary school children living in and around Kuala Lumpur, Malaysia, revealed that 12.9% of the children were infested with Pediculus humanus capitis. Indians (28.3%) and Malays (18.9%) have a higher prevalence than Chinese (4.6%). The higher prevalence among Indians and Malays correlates well with their lower socio-economic status in the community; long hair also contributes to the higher rates of pediculosis among them. The prevalence rate was found to be related to socio-economic status, length of hair, family size, age, crowding and personal hygiene. Treatment with 0.2% and 0.5% malathion in coconut oil gave cure rates of 93% and 100%. Treatment with gammexane and actellic at 0.5% concentration gave a cure rate of 100% against adults and nymphs of Pediculus humanus capitis.
DDT indoor residual sprayings in the Province of Central Java, Indonesia, for the past 28 years are reoieuied in terms ofAnopheles aconitus resistance to DDT and incidences of malaria. DDT resistance by An. aconitus has continued to increase and spread since its first detection in Central Java in 1962. In most areas malaria transmission is perennial, which appears to be not interrupted by DDT residual spraying of inner walls because of DDT resistance. In searching for alternatives to DDT, fenitrothion was the most effective residual compound among five insecticides tested in village scale trials, giving control for about 13 weeks at 2 g/m2. Because the target species rests largely on the lower portions of walls, when applied at 2 g/m2 to only one horizontal swath between 10 and 85 cm from the ground, this compound was effective for over two months, about one-third less than that found in full coverage applications. For a single application this selective method reduced the amount of insecticide by 65 percent and manpower by 43 percent, and three applications, to compensate for less residual effectiveness, will reduce insecticide use by about 50 percent and manpower by 15 percent, compared to two applications with full coverage. Further field investigations on application of insecticides and the ecology of the DDT-resistant vector continue to be needed in Central Java to reduce further the cost of residual sprayings in malaria control programmes. This review also suggests the possibility of other selective spraying methods and anti-vector methods applicable at the village level by the community.
Insecticides Abate, DDT, Dowco-214, Dursban, fenitrothion, fenthion, gamma-HCH, and malathion were tested against the field collected fourth instars larvae of Mansonia from Penang Island, Malaysia. The larvae appeared to be highly susceptible to Dursban and Abate with LC50 values of 1.54 and 1.92 parts per billion respectively. Other chemicals, in decreasing degree of effectiveness, were gamma-HCH, fenthion, P,P'-DDT, Dowco-214, fenithrothion and malathion. The potential use of these chemicals in Mansonia control was discussed. A simple method for collecting and testing Mansonia larvae was also described.
Toxicological studies of four insecticides (malathion, carbaryl, bioresmethrin, and GH 74) against Musca domestica vicinia (Ampang strain) were undertaken with particular reference to age, sex and posttreatment temperature. It was found that bioresmethrin and GH 74, both with a negative temperature coefficient, have great potential for use against houseflies. In vitro inhibitory studies of head and body esterases showed that unlike malathion and carbaryl, bioresmethrin had only negligible effect on these enzymes. The possibilities of using bioresmethrin and GH 74 for controlling the housefly problem in the Cameron Highlands, West Malaysia are discussed.