Development of a controlled release formulation of an indigenous insect growth regulator, DPE-28, a substituted diphenylether, for controlling the breeding of Culex quinquefasciatus.

Background & objectives: DPE-28, a substituted diphenyl ether (2,6-ditertiarybutyl phenyl-2’,4’-dinitro phenyl ether) was reported to exhibit promising insect growth regulating activity against Culex quinquefasciatus, the vector of lymphatic filariasis. A controlled release formulation (CRF) of DPE-28 has been developed to control Cx. quinquefasciatus in its breeding habitats. Toxicity of DPE-28, safety to non-target mosquito predators and the release profile of the CRF of DPE-28 are studied and discussed. Methods: The acute oral and dermal toxicity was tested in male and female Wistar rats as per the Organization for Economic Cooperation and Development (OECD) guidelines 425 and 402 respectively. The toxicity of DPE-28 to non-target predators was tested as per the reported procedure from this laboratory. The CRF of DPE-28 was prepared by following the reported procedure developed at this laboratory earlier. The concentration of DPE-28 released from the CRF was monitored by HPLC by constructing a calibration graph by plotting the peak area in the Y-axis and the concentration of DPE-28 in the X-axis. Results: DPE-28 has been tested for acute oral toxicity and found to be moderately toxic with LD50 value of 1098 mg/kg body weight (b.w). The results of the acute dermal toxicity and skin irritation studies reveal that DPE-28 is safe and non-irritant. DPE-28 when tested at 0.4 mg/litre against non-target mosquito predators did not produce any mortality. The release profile of the active ingredient DPE-28 from the CRF by HPLC technique showed that the average daily release (ADR) of DPE-28 ranged from 0.07 to 5.0 mg/litre during first four weeks. Thereafter the matrix started eroding and the ADR ranged from 5 to 11 mg/litre during the remaining 5 wk. The cumulative release of active ingredient showed that > 90 per cent of the active ingredient was released from the matrix. Interpretation & conclusions: The controlled release matrix of DPE-28 was thus found to inhibit the adult emergence (>80%) of Cx. quinquefasciatus for a period of nine weeks. The CRF of DPE-28 may play a useful role in field and may be recommended for mosquito control programme after evaluating the same under field conditions.

Lethal and Sublethal Effects of Methoxyfenozide on the Development, Survival and Reproduction of the Fall Armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae)

The lethal and sublethal effects of the ecdysone agonist methoxyfenozide on the fall armyworm, Spodoptera frugiperda (J. E. Smith), were investigated by feeding a methoxyfenozide-treated diet to fifth instars until pupation in doses corresponding to the LC10 and LC25 for the compound. Larval mortality reached 8 percent and 26 percent in the low and high concentration groups, respectively, on the seventh day of the experiment. A progressive larval mortality of 12 percent for the LC10 and 60 percent for the LC25 was observed before pupation. Treated larvae exhibited lower pupal weights, higher pupal mortality, presence of deformed pupae, and more deformed adults than untreated larvae. The incorporation of methoxyfenozide into the diet had a significant effect on the timing of larval development. The development period for males and females was about seven days longer than the controls for both concentrations tested. In contrast, the compound affected neither pupae nor adult longevity. Finally, S. frugiperda adults that resulted from fifth instars treated with methoxyfenozide were not affected in their mean cumulative number of eggs laid per female (fecundity), nor percentages of eggs hatched (fertility), or the sex ratio. Our results suggest that the combination of lethal and sublethal effects of methoxyfenozide may have important implications for the population dynamics of the fall armyworm.

Susceptibility of Aedes aegypti (L) to the insect growth regulators diflubenzuron and methoprene in Uberlândia, State of Minas Gerais

Aedes aegypti (L) (Diptera: Culicidae) was reared in several concentrations of diflubenzuron and methoprene under laboratory conditions in Uberlândia, State of Minas Gerais, southeastern Brazil. Characteristics such as LC50 and LC95, the susceptibility of immature stages of different ages to these insect growth regulators and their residual effects were studied. The LC50 and LC95 of diflubenzuron and methoprene were 5.19 and 12.24 ppb; 19.95 and 72.08 ppb, respectively. While diflubenzuron caused great mortality in all larval instars, methoprene was more effective when the mosquito was exposed from the start of the fourth larval instar onwards. Commercial concentrations of these two insect growth regulators close to LC95 presented greater residual activity than did their respective technical formulations. The parameters were compared with those obtained elsewhere. The characteristics investigated here indicate that these insect growth regulators are effective alternatives for controlling the dengue vector in the Uberlândia region.

Aedes aegypti (L) (Diptera: Culicidae) foi criado em várias concentrações de diflubenzuron e methoprene sob condições de laboratório em Uberlândia, Minas Gerais, sudeste do Brasil. Foram estudados aspectos tais como, CL50 e CL95, suscetibilidade de estágios imaturos de diferentes idades a estes insect growth regulators e seu efeito residual. As CL50 e CL95 de diflubenzuron e methoprene foram: 5,19 e 12,24ppb; 19,95 e 72,08ppb, respectivamente. Enquanto diflubenzuron causou grande mortalidade em todos os estádios larvais, methoprene causou maior mortalidade quando o mosquito foi exposto a partir do início do quarto estádio larval. As concentrações comerciais dos dois insect growth regulators próximas às CL95 mostraram maior atividade residual que suas respectivas formulações técnicas. Os parâmetros são comparados com aqueles obtidos em outros locais. Os aspectos aqui investigados indicam estes insect growth regulators como alternativas efetivas para o controle do vetor da dengue na região de Uberlândia.

Persistência e eficácia do regulador de crescimento pyriproxyfen em condicões de laboratório para Aedes aegypti / Persistence and efficacy of growth regulator pyriproxyfen in laboratory conditions for Aedes aegypti

A persistência e a eficácia do regulador de crescimento pyriproxyfen foram testadas em concentracões de 0,01 e 0,05ppm, contra larvas de Aedes aegypti, utilizando os recipientes caixas d'água (45 litros), frascos de vidro (5 litros) e baldes de plástico (20 litros). As avaliacões foram nos dias 1, 7, 15, 30, 45, 60, 90 e 120 após o tratamento usando larvas de 3º e 4º estádio de Aedes aegypti. Foi calculado o percentual de mortalidade de larvas, pupas e adultos, percentual de inibicão de emergência de adulto e duracão dos bioensaios. Observou-se que a persistência foi de 45 dias e 90 dias para concentracão final de 0,01 e 0,05ppm de pyriproxyfen, respectivamente. Observamos que a mortalidade de pupas foi significativamente maior que a de larvas e de adultos para todos os recipientes e concentracões.

Heterochrony in cotton bollworm, Earias vitella (Fb) following ethoxyprecocene administration.

Administration of a synthetic precocene analogue, 7-ethoxy-6-methoxy-2,2-dimethyl chromene to fourth and last instar larvae of E. vitella results into heterochrony, viz. prothetely and metathetely. These disturbances are due to interference with the endocrine system and application of juvenile hormone to treated larvae abolishes the effect of ethoxyprecocene.

Macromolecular synthesis in wing discs of Spodoptera mauritia Boisd: effects of a juvenile hormone analogue.

Last instar larvae of S. mauritia treated topically on day 0, day 1, day 2 and day 3 with a daily (dose of 25 microg juvenile hormone analogue (JHA) moulted into supernumerary larvae. The imaginal discs of the supernumerary larvae especially those of mouthparts and thoracic appendages showed pupal characteristics. However the wing discs, which showed only partial differentiation, were uneverted and highly tanned. In an effort to provide an explanation to this anomaly the RNA, DNA and protein profile in the wing discs of supernumerary larvae were studied. Quantitative analysis of DNA, RNA and protein showed a considerable increase in the amount of DNA and protein and a decline in RNA level. SDS-PAGE analysis of wing disc proteins of JHA treated larvae showed a reduction in the expression of many major proteins that were predominant in the wing discs of control larvae. The results suggest that JHA induced inactivation of genes involved in the synthesis of proteins needed for evagination process may be responsible for the formation of uneverted, partially differentiated pupal wing discs in supernumerary larvae.

Hormonal implication in Bracon-venom-induced paralysation of the host larva of Corcyra cephalonica (Lepidoptera: Pyralidae).

To facilitate oviposition, the ectoparasite Bracon hebetor, injects its venom, a paralysing toxin, to the host Corcyra larva that ultimately dies without showing any metamorphic change, even if allowed to remain unparasitised. At the initial stage of venom injection the rate of heartbeat of the host becomes abruptly high. This has been explained from the synergistic action of the substances of poison gland and calyx. The paralysed larvae subsequent to envenomization die within 240 hr. Application of hydroprene as single dose or with a booster dose after paralysation mostly increases the survival period considering heart beat as the index. The predicted value of survival period (714.4 hr), determined from a fitted equation obtained from the relationship between heart beat and survival period, indicates that a 100 microg treatment/larva with a booster dose of 50 microg/larva most effectively lengthens the period. It is concluded that the venom-induced physiological dysfunction of the immobilised larvae, as indicated in the rate of heart beat and survival period, though can be recovered to some extent after the application of juvenoids, there cannot occur any metamorphic change of these larvae. The parasitoid, therefore, succeeds in completing its development and metamorphosis by arresting the development of its host through an indirect hormonal suppression. The findings indicate an endocrine implication in host-parasite relationship in insect.

Inhibitory effect of 1-benzyl-5[(C)-2,6-dimethyl-1,5-heptadienyl] imidazole (KK-42) on desert locust, Schistocerca gregaria forsk.

Imidazole compound KK-42 was tested for its inhibitory effect on Schistocerca gregaria. The compound caused 80% mortality after 19 days of treatment with 100 microg of KK-42/insect. It also caused the production of deformed adults at even lower doses. Ecdysteroid content, when estimated by RIA, declined due to KK-42 treatment in both haemolymph and ovaries of the insect. Reduction in ecdysteroid titer was maximum at 100 microg of KK-42/insect. The value estimated by RIA was 0.34 ng/pL haemolymph as compared to 0.97 ng/microL in case of controls. Similarly,ecdysteroid content of ovaries detected was 0.48 ng/mg in treated ones as compared to 0.61 ng/mg in controls. Histological studies, however, revealed no alteration in the ovarian tissues due to KK-42 treatment.

Effects of a juvenoid, DPE-28, on biology and behaviour of Culex quinquefasciatus, the human filariasis vector.

Studies on the effect of a juvenoid, DPE-28 (2,4-dinitrophenyl-2',6'-di-tertiarybutyl phenyl ether) on biology and behaviour of Cx. quinquefasciatus showed that the developmental duration, sex ratio, mating success and blood feeding were considerably affected by the exposure of larvae and pupae to the compound. Exposure of fourth instar larvae to 0.007 (EI90) and 0.0019 (EI50) ppm of DPE-28 prolonged the duration of pupation by 58.6 and 52.4 hr and delayed the adult emergence by 35.4 and 17.7 hr in males and 36.8 and 21.1 hr in females respectively. Exposure of freshly ecdysed pupae to 10 and 5 ppm delayed the adult emergence with respect to the control by 54.3 and 32.4 hr in males and 55.2 and 33.2 hr in females respectively. The sex ratio of the adults emerged from treated larvae and pupae was also affected. The female mosquitoes that survived from the exposed fourth instar larvae and pupae exhibited a low blood engorgement ratio. This depression in blood feeding was more pronounced in adults emerged from treated pupae than that of treated fourth instar larvae. A significant proportion of adults emerged from treated larvae and pupae were able to feed only partially. Mating success of the treated populations declined considerably when crosses were made between the males and females emerged from treated fourth instar larvae and pupae. The adults emerged from treated larvae and pupae showed a significant reduction in the oviposition.

Morphogenetic derangements in the reproductive system of Bracon hebetor, a beneficial parasitoid, bred on juvenoid treated host (Corcyra cephalonica) larvae.

Effect of juvenoids (hydroprene and methoprene) on the ecto-parasite B. hebetor was investigated by rearing them upon the juvenoid treated ultimate instar host larvae of C. cephalonica. Emerged adultoid wasps of either sexes obtained from treated series showed anatomical deformities in the reproductive systems. Ill-developed ovaries with reduced length, terminally free ovarioles and abnormal testicular growth showing non-fusion of lobes were the important abnormal features. Data on measurements of male reproductive system, e.g., width (transverse axis) of testis, length of common vas deferens plus ejaculatory duct and length of accessory gland showed significant difference (P < 0.05) from control.

Effect of fumigation and juvenile hormone on oxygen consumption of the larvae of Corcyra cephalonica (Stainton).

Reduced effect of JH-I, JH-II and JH-III on oxygen consumption of II-V instars and increased effect on the oxygen consumption of VI instar larvae suggested that control of corcyra with juvenile hormones could be brought about only when applied to just emerged VI instar larvae. Similarly fumigation of juvenile hormone treated larvae could prove beneficial only at VI instar stage. The possibility of enhancing the effect of fumigant with pretreatment of JH will be futile as even only JH treated VI instar larvae develop into abnormal individuals which die later.

Development of Wuchereria Bancrofti in culex pipiens L. [Diptera: Culicidae] exposed in the larval instar to sublethal dosages of insecticides and one insect growth regulator and their influence on reproduction of filaria-infected mosquitoes

The effects of exposure of Culex pipiens larvae to sublethal concentrations of larvicides on uptake and development of Wuchereria bancrofti, survival rate and reproduction of filaria- infected mosquitos were investigated. The overall infection and infective rates were significantly reduced in mosquitos treated with Triton X-100, either alone or combined with permethrin. All larvicides reduced the number of infective larvae [L3]/mosquito and induced deformities among the different parasite stages. Addition of Triton X-100 to DPX or to permethrin significantly reduced egg Production

Effect of juvenile hormone and ecdysone on initiating blood meal-dependent vitellogenic ovarian cycles in autogenous and anautogenous culex pipiens [Diptera: culicidae]

The effect of juvenile hormone [JH] and ecdysone on initiating blood meal - stimulated ovarian cycles was studied in autogenous [2[nd] ovarian cycle] and anautogenous [1[st] ovarian cycle] Cx. pipiens. Although vitellogenesis was initiated in both mosquitoes in response to ecdysone, JHI and sequential JHI-ecdysone treatments induced vitellogenesis in anautogenous females only. JHI treatment initiated vitellogenesis in Ca. 30% of anautogenous females and only half of those were able to form mature eggs. Sequential JHI- ecdysone treatment triggered vitellogenesis in 10 out of 25 anautogenous females and mature eggs were formed in three females only. In response to ecdysone, vitellogenesis in 10 out of 18 autogenous females and mature eggs were formed in two individuals. In Ca. 70% of anautogenous siblings vitellogenesis was initiated and in most of those females mature eggs were developed. Our data suggests that differences in hormone - dependent events of vitellogenesis in autogenous and anautogenous Cx. pipiens seem to be related to the regulation of ecdysteroide release

Cyclopcidal activity of some new insect growth regulators.

A series of synthetic terpenoid diethers, with known mosquito development inhibitory activity, were tested for cyclopcidal activity against adult cyclops, Mesocyclops leuckarti. Four among these exhibited promising activity comparable to that of the conventional organophosphate, temephos, thus providing for the first time a possible reserve arsenal for the control of the guineaworm disease vector.

Hormonal regulation of macromolecular synthesis in testes and accessory reproductive glands of Spodoptera litura during post-embryonic and adult development.

In S. litura testicular growth during the last larval instar and early pupal stage is associated with significant increase in DNA, RNA and protein contents. DNA synthesis is stimulated by 20-hydroxyecdysone (20-HE) in the penultimate instar testes. 20-HE injection in ligated late last instars increases the testicular weight and protein content. Accessory reproductive gland (ARG) development takes place during the mid and late pupal stages. Protein synthesis in the pharate adult ARG is stimulated by 20-HE. Juvenile hormone has no effect on ARG protein synthesis.

Effects of cyromazin and methoprene on the developmental stages of Anopheles dirus, Aedes aegypti and Culex quinquefasciatus (Diptera : Culicidae).

The effects of two chemical compounds, cyromazin and methoprene, on the developmental stages of Anopheles dirus, Aedes aegypti and Culex quinquefasciatus were investigated under laboratory conditions, with the mean temperature of 24 degrees +/- 1 degree C and the relative humidity at 65-75%. Both compounds were tested against the second, third and fourth instar larvae. The concentrations of cyromazin used for An. dirus and Cx. quinquefasciatus ranged from 0.0008 to 0.5 mg/l; and for Ae. aegypti from 0.004 to 2.5 mg/l. The concentrations of methoprene used for An. dirus, Ae. aegypti and Cx. quinquefasciatus ranged from 0.00016 to 0.1 mg/l. The mortality rates were found to be relatively high in larval and pupal stages when treated with cyromazin and methoprene. The primary toxic effects of cyromazin were on the second stage larvae. The LC50 values for cyromazin on the second, third and fourth stage larvae were, respectively, 0.0027, 0.0042 and 0.0114 mg/l for An. dirus, and 0.1662, 0.2307 and 0.3005 mg/l for Ae. aegypti. Cx. quinquefasciatus was the most sensitive species to cyromazin with LC50 values for second, third and fourth stage larvae of 0.0015, 0.0068 and 0.0130 mg/l, respectively. The primary toxic effects of methoprene were in the fourth stage larvae. The LC50 values for methoprene on the second, third and fourth stage larvae were, respectively, 0.0110, 0.0041 and 0.0022 mg/l for An. dirus, and 0.0077, 0.0034 and 0.0025 mg/l for Ae. aegypti. Cx. quinquefasciatus was the most sensitive species to methoprene, with LC50 values for second, third and fourth stage larvae of 0.0013, 0.0008 and 0.0006 mg/l, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of a juvenile hormone analogue on the morphology and biology of Aedes scutellaris malayensis Colless (Diptera: Culicidae).

Effects of 2-methyl-9(4-isopropylphenyl)-2,6-dimethyl nonane, or MV-678, a juvenile hormone analogue, on Aedes scutellaris malayensis Colless were investigated under laboratory conditions (29 degrees +/- 2 degrees C and 86 +/- 3% RH). The MV-678 was tested against the first, second, third and fourth instar larvae. The concentrations used were 0.0032, 0.016, 0.08, 0.4 and 2.0 mg/l. The morphogenetic aberrations were determined and divided into 8 groups, among which they included (1) death larvae, (2) late fourth instar larvae before pupation, (3) larvae with pupae partly emerged, (4) white pupae, (5) brown pupae, (6) elephantoid pupae of which pupae with adults visible inside, (7) pupae with apparently adults partly emerged and (8) death adults. The percentage mortality rates were found to be relatively high in pupal and larval stages when they were treated with 2.0 mg/l. The LC50 values were 0.26, 0.175, 0.06 and 0.032 mg/l for the first, second, third and fourth instar larvae respectively. The effectiveness of MV-678 at 2.0 mg/l was about 11 days under the open air conditions (28 degrees +/- 2 degrees C and 72 +/- 3% RH). When the fourth instar larvae were treated with 0.4 and 2.0 mg/l of MV-678, the LC50 values were 3.1 and 7.1 days respectively.