RNAi-mediated knockdown of SPOOK reduces ecdysteroid titers and causes precocious metamorphosis in the desert locust Schistocerca gregaria.

The Halloween gene SPOOK (SPO) is involved in the production of the active metabolite of ecdysteroid, 20-hydroxyecdysone (20E), in insects. A previous study showed that RNAi-mediated knockdown of SPO in Schistocerca gregaria last instar nymphs markedly reduced the hemolymph 20E titer, but did not affect metamorphosis. In the present study, the effects of SPO interference on development were re-examined in this locust. Injections of SPO double-stranded RNA (dsSPO) into nymphs at mid and late instars significantly delayed nymphal development and interfered with molting. The 20E levels of dsSPO-treated nymphs were generally low, with a delayed, small peak, suggesting that disturbance of the 20E levels caused the above developmental abnormalities. A small proportion of the dsSPO-injected nymphs metamorphosed precociously, producing adults and adultoids. Precocious adults were characterized by small body size, short wings with abbreviated venation, and normal reproductive activity. Fourth instar nymphs that precociously metamorphosed at the following instar exhibited temporal expression patterns of ecdysone-induced protein 93F and the juvenile hormone (JH) early-inducible gene Krüppel homolog 1 similar to those observed at the last instar in normal nymphs. Adultoids displayed mating behavior and adultoid females developed eggs, but never laid eggs. JH injection around the expected time of the 20E peak in the dsSPO-injected nymphs completely inhibited the appearance of adultoids, suggesting that appearance of adultoids might be due to a reduced titer of JH rather than of 20E. These results suggest that SPO plays an important role in controlling morphogenesis, metamorphosis, and reproduction in S. gregaria.

Toxicity and resistance of field collected Musca domestica (Diptera: Muscidae) against insect growth regulator insecticides.

The house fly, Musca domestica, is a serious pest of public health importance with the ability to develop insecticide resistance. The focus of the present study was to evaluate toxicity and resistance of the field collected house flies from Punjab, Pakistan, against insect growth regulator (IGR) insecticides. House flies collected from six different localities exhibited very low levels of resistance to cyromazine, triflumuron, and methoxyfenozide compared with the Lab-susceptible reference strain, with resistance ratios (RRs) ranging between 3.56- to 8.19-fold, 1.45- to 3.68-fold, and 2.20- to 8.60-fold, respectively. However, very low to low levels of resistance were observed for pyriproxyfen and very low to moderate levels for lufenuron with RRs ranged from 4.13- to 11.63-fold to 8.57- to 22.75-fold, respectively. There was a significant correlation between RRs of cyromazine and triflumuron (r = 0.976, p < 0.001), suggesting the possibility for cross-resistance. Susceptibility status of different IGRs in Pakistani strains of house flies is reported here for the first time. The trend observed in decreasing susceptibility towards different IGR insecticides tested will continue unless resistance management practices are followed.

C3H/He Mice as an Incompatible Cholangiocarcinoma Model by Clonorchis sinensis, Dicyclanil and N-Nitrosodimethylamine.

Clonorchis sinensis is a Group-I bio-carcinogen, associated with cholangiocarcinoma (CCA). The hamster is the only experimental model of C. sinensis-mediated CCA, but we oblige another animal model. The present study intended to develop a C. sinensis (Cs) mediated CCA model using C3H/He mice, co-stimulated with N-nitrosodimethyl-amine (NDMA) and dicyclanil (DC). The mice were divided into 8 groups with different combinations of Cs, NDMA, and DC. Six months later the mice were sacrificed and subjected to gross and histopathological examination. The body weights were significantly reduced among the groups treated with 2 or more agents (eg. Cs+NDMA, Cs+DC, NDMA+DC, and Cs+NDMA+DC). In contrast, liver weight percentages to body weight were increased in above groups by 4.1% to 4.7%. A Change of the spleen weight was observed only in Cs+NDMA group. Though C. sinensis infection is evident from hyperplastic changes, only 1 worm was recovered. T wo mice, 1 from Cs and the other from Cs+DC group, showed mass forming lesions; 1 (281.2 mm(3)) from the Cs group was a hepatocellular adenoma and the other (280.6 mm(3)) from the Cs+DC group was a cystic mass (peliosis). Higher prevalence of gray-white nodules was observed in Cs group (42.9%) followed by Cs+NDMA+DC group (21.4%). The mice of the Cs+NDMA+DC group showed hyper-proliferation of the bile duct with fibrotic changes. No characteristic change for CCA was recognized in any of the groups. In conclusion, C3H/He mice produce no CCA but extensive fibrosis when they are challenged by Cs, NDMA, and DC together.

Timing Spring Insecticide Applications to Target both Amyelois transitella (Lepidoptera: Pyralidae) and Anarsia lineatella (Lepidoptera: Gelechiidae) in Almond Orchards.

Amyelois transitella (Walker) (Lepidoptera: Pyralidae) and Anarsia lineatella Zeller (Lepidoptera: Gelechiidae) are key Lepidoptera pests of almonds in California. Spring insecticide applications (early to mid-May) targeting either insect were not usually recommended because of the potential to disrupt natural enemies when broad-spectrum organophosphates and pyrethroids were applied. The registration of reduced risk compounds such as chlorantraniliprole, methoxyfenozide, and spinetoram, which have a higher margin of safety for natural enemies, makes spring (early to mid-May) application an acceptable control approach. We examined the efficacy of methoxyfenozide, spinetoram, and chlorantraniliprole at three spring application timings including the optimum spring timing for both A. lineatella and A. transitella in California almonds. Our study also examined the possibility of reducing larval populations of A. lineatella and A. transitella simultaneously with a single spring insecticide application. There were no significant differences in the field efficacy of insecticides targeting either A. lineatella or A. transitella, depending on application timing for the three spring timings examined in this study. In most years (2009-2011), all three timings for each compound resulted in significantly less A. transitella and A. lineatella damage when compared with an untreated control, though there was some variation in efficacy between the two species. Early to mid-May applications of the reduced-risk insecticides chlorantraniliprole and spinetoram can be used to simultaneously target A. transitella and A. lineatella with similar results across the potential timings.

Control of Aedes aegypti and Ae. albopictus, the vectors of dengue and chikungunya, by using pheromone C21 with an insect growth regulator: Results of multicentric trials from 2007-12 in India.

BACKGROUND & OBJECTIVES: Aedes mosquito control has gained much importance nowadays in view of rise in number of reported cases of dengue and chikungunya in India and other countries. In the present study, C21 attracticide (containing a pheromone and an insect growth regulator­IGR, developed by Defence Research and Development Establishment (DRDE), Gwalior, India was tested for its feasibility for surveillance and control of Aedes mosquito in a multicentric mode from October 2007 to June 2012 in urban (Delhi, and Bengaluru district, Karnataka) and suburban (Alappuzha district, Kerala) settings of the country in three phases. METHODS: Across the randomly selected households in each study area, two to four containers treated with attracticide (experimental) and untreated (control) were placed and monitored by trained surveillance workers on weekly/ fortnightly basis for determining the presence of eggs, larvae and pupae. Container positivity, percent larvae, egg and pupae collected were determined during different phases and analyzed statistically using SPSS 18.0. RESULTS: Container positivity was found statistically significant at Bengaluru and Alappuzha, Kerala while in Delhi, it was found non-significant. Eggs collected from experimental containers were significantly higher in comparison to control at all the locations except Delhi. Also larvae collected from control containers were significantly higher at all the locations except Bengaluru. Pupae collected from control containers remained significantly higher at all the locations as no pupal formation was recorded from experimental containers. INTERPRETATION & CONCLUSION: The use of C21 attracticide hampered pupal formation, thus inhibiting adult population in the study areas. The study established that C21 attracticide was efficacious in the field conditions and has potential for use in surveillance and management of dengue and chikungunya mosquitoes.

Exploring new thermal fog and ultra-low volume technologies to improve indoor control of the dengue vector, Aedes aegypti (Diptera: Culicidae).

Control of the mosquito vector, Aedes aegypti (L.), inside human habitations must be performed quickly and efficiently to reduce the risk of transmission during dengue outbreaks. As part of abroad study to assess the efficacy of dengue vector control tools for the U.S. Military, two pesticide delivery systems (ultra-low volume [ULV] and thermal fog) were evaluated for their ability to provide immediate control of Ae. aegypti mosquitoes with a contact insecticide inside simulated urban structures. An insect growth regulator was also applied to determine how well each sprayer delivered lethal doses of active ingredient to indoor water containers for pupal control. Mortality of caged Ae. aegypti, pesticide droplet size, and droplet deposition were recorded after applications. In addition, larval and pupal mortality was measured from treated water samples for 4 wk after the applications. The ULV and the thermal fogger performed equally well in delivering lethal doses of adulticide throughout the structures. The ULV resulted in greater larval mortality and adult emergence inhibition in the water containers for a longer period than the thermal fogger. Therefore, the ULV technology is expected to be a better tool for sustained vector suppression when combined with an effective insect growth regulator. However, during a dengue outbreak, either delivery system should provide an immediate knockdown of vector populations that may lower the risk of infection and allow other suppression strategies to be implemented.

Attractive toxic sugar baits mixed with pyriproxyfen sprayed on plants against adult and larval Aedes albopictus (Diptera: Culicidae).

The effect of spraying a mixture of the insect growth regulator (IGR) pyriproxyfen (1 mg/liter) and either 1% boric acid sugar bait or eugenol sugar bait on croton petra plants (Codiaeum variegatum L.) was evaluated against the container-inhabiting mosquito, Aedes albopictus (Skuse). Treatments were applied to plants and evaluated against adult and larval Ae. albopictus in the laboratory through contact and wash off experiments, respectively. The control treatment lacked an active ingredient and were treated with an attractive sugar bait. The plants treated with attractive toxic sugar baits plus the IGR resulted in 60-100% mortality of laboratory-reared adult Ae. albopictus. The pyriproxyfen solutions collected from the plant wash experiment resulted in 80-100% emergence inhibition to the exposed third- and fourth-instar larvae, compared with the untreated control. Attractive toxic sugar baits mixed with the IGR not only provide effective control of adult mosquitoes, but also provide additional control of larval mosquitoes after being washed off from the treated plants.

Pyriproxyfen and house flies (Diptera: Muscidae): effects of direct exposure and autodissemination to larval habitats.

Pyriproxyfen is an insect growth regulator with juvenile hormone-like activity that has potential uses for dipterans that are difficult to manage with conventional insecticides, such as house flies (Musca domestica L.). The objectives of this study were to determine the efficacy of this insect growth regulator against house flies using variety of delivery systems and target life stages, including an evaluation of the potential for autodissemination by female flies to larval development sites. Adult female house flies exposed to filter paper (3.75% active ingredient) or sugar treated with pyriproxyfen (0.01-0.1%) produced significantly fewer F1 pupae than untreated flies. Adult emergence from pupae was unaffected. In contrast, treatment of larval rearing medium with 0.35 ml/cm2 of a 12 mg pyriproxyfen/liter preparation had no effect on the number of pupae developing from eggs but markedly inhibited adult emergence from those pupae. There was little difference in susceptibility between an insecticide-susceptible and a wild strain of house fly. The LC50 for inhibiting fly emergence of dust formulations in diatomaceous earth incorporating commercial pyriproxyfen products ranged from 8 to 26 mg/liter, with little difference among products. Compared with untreated flies, significantly fewer pupae were produced at concentrations > 0.5% and no adults were produced at concentrations > 0.05% pyriproxyfen. When gravid females were exposed for 1 h to treated fabric (6 mg pyriproxyfen/cm2) and allowed to oviposit in rearing media containing eggs, sufficient pyriproxyfen was autodisseminated to reduce adult emergence from those eggs by > 99%. Intermittent contact with treated fabric over 2 d reduced adult emergence by 63-76%.

Efficacy of pyriproxyfen-treated nets in sterilizing and shortening the longevity of Anopheles gambiae (Diptera: Culicidae).

Pyrethroid-resistant malaria vectors have become a serious threat for malaria control, and bed nets that reduce the development of resistance are urgently needed. Here, we tested the effects of bed nets treated with the insect growth regulator pyriproxyfen against adult female Anopheles gambiae Giles (Diptera: Culicidae) under laboratory conditions. Noninsecticidal nets made of 195 denier monofilament polyethylene with a mesh size of 75 holes per square inch (equivalent to the Olyset Net) were dipped in a 0.1, 0.01, or 0.001% (wt:vol) alcohol solution of pyriproxyfen and dried overnight. Adult females of an insecticide-susceptible An. gambiae strain were exposed to treated and untreated nets before and after a bloodmeal. Bioassays showed that females were completely sterilized after exposure to 0.1% (35 mg [AI]/m2) and 0.01% pyriproxyfen-treated nets both before and after a bloodmeal. In addition, adult longevity decreased after exposure to the pyriproxyfen-treated nets in a concentration-dependent manner. The sterilizing and life-shortening effects of pyriproxyfen on the vector mosquito indicate that the combined use of pyriproxyfen and pyrethroids on bed nets has the potential to provide better malaria control and prevent the further development of pyrethroid resistance in malaria vectors.

An autodissemination station for the transfer of an insect growth regulator to mosquito oviposition sites.

A prototype autodissemination station to topically contaminate oviposition-seeking container-dwelling mosquitoes with the insect growth regulator, pyriproxyfen, was developed and tested in the laboratory. Our test subject was the Asian tiger mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae), an urban species that colonizes small-volume cryptic larval habitats and is a skip ovipositor that visits multiple containers. The station consists of a water reservoir to attract gravid females, which is joined to a transfer chamber designed to contaminate visiting mosquitoes. The unit is easily constructed by moulding wet shredded cardboard using corn starch as a binder. The essential criteria that must be met to prove the efficacy of an autodissemination station require it to demonstrate effectiveness in attracting the target insect, in transferring the toxicant to the insect that will disperse the agent, and in facilitating the subsequent transfer of the toxicant from the insect to target habitats at a lethal concentration. Cage experiments demonstrated that the unit was readily accepted by gravid females as an oviposition site. A powder formulation of pyriproxyfen-impregnated silica particles adhered to visiting Ae. albopictus females (mean: 66 particles/female), although particles were lost over time. In cage (2.2 m(3) ) trials, pyriproxyfen-charged stations resulted in 100% inhibition of adult emergence, whereas in small-room (31.1 m(3) ) trials, 81% emergence inhibition was recorded. The venereal transfer of pyriproxyfen from contaminated males to virgin females was also observed, and pyriproxyfen was subsequently transferred to water-holding containers at concentrations that inhibited emergence. Key autodissemination station features include lack of maintenance requirements, biodegradable construction, low cost and low risk.

Improved chemical control for the Mexican rice borer (Lepidoptera Crambidae) in sugarcane: larval exposure, a novel scouting method, and efficacy of a single aerial insecticide application.

A three-treatment aerial application insecticide experiment was conducted in five commercial sugarcane, Saccharum spp., fields in south Texas to evaluate the use of pheromone traps for improving chemical control of the Mexican rice borer, Eoreuma loftini (Dyar), in 2009 and 2010. A threshold of 20 moths/trap/wk was used to initiate monitoring for larval infestations. The percentage of stalks with larvae on plant surfaces was directly related to the number of moths trapped. Reductions in borer injury and adult emergence were detected when a threshold of >5% of stalks with larvae present on plant surfaces was used to trigger insecticide applications. Novaluron provided superior control compared with beta-cyfluthrin; novaluron treated plots were associated with a 14% increase in sugar production. A greenhouse experiment investigating establishment and behavior of E. loftini larvae on two phenological stages of stalkborer resistant, HoCP 85-845, and susceptible, HoCP 00-950, sugarcane cultivars determined that more than half of larvae on HoCP 00-950 and > 25% on HoCP 85-845 tunneled inside leaf mid-ribs within 1 d of eclosion, protected therein from biological and chemical control tactics. Exposure time of larvae averaged < 1 wk for all treatments and was shortest on immature HoCP 00-950 and longest on mature HoCP 85-845. This study shows a short window of vulnerability of E. loftini larvae to insecticide applications, and demonstrates the potential utility of pheromone traps for improving insecticide intervention timing such that a single properly timed application may be all that is required.

Evaluation of juvenile hormone analogues as rodent feed-through insecticides for control of immature phlebotomine sandflies.

The juvenile hormone analogues methoprene and pyriproxyfen were evaluated as rodent feed-through insecticides for control of immature stages of the sandfly Phlebotomus papatasi Scopoli (Diptera: Psychodidae). The development and survival of P. papatasi second-instar larvae fed faeces from Syrian hamsters, Mesocricetus auratus, that had been fed a diet containing methoprene (0, 9.788, 97.88 or 978.8 p.p.m.) or pyriproxyfen (0, 9.82, 98.2 or 982 p.p.m.) were evaluated. The faeces of methoprene-treated hamsters greatly reduced the percentage of larvae that pupated at all concentrations tested and prevented adult emergence at all but the lowest concentration (9.788 p.p.m.). Pyriproxyfen prevented both pupation and adult emergence at all concentrations tested. The results of this study suggest that a control strategy using rodent baits containing juvenile hormone analogues to control phlebotomine sandflies that live in rodent burrows and feed on rodent faeces may be possible. As rodent reservoirs and vectors of Leishmania major live in close association in many parts of the Middle East, control of the transmission of the agent of zoonotic cutaneous leishmaniasis may also be possible.

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 LD 50 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.

Areawide field study on effect of three chitin synthesis inhibitor baits on populations of Coptotermes formosanus and Reticulitermes flavipes (Isoptera: Rhinotermitidae).

Periodic sampling of 43 independent monitors, initially active with Formosan subterranean termite, Coptotermes formosanus Shiraki, or the eastern subterranean termite, Reticulitermes flavipes (Kollar) (Isoptera: Rhinotermitidae), was conducted to evaluate the effects of cellulose baits containing one of three chitin synthesis inhibitors (CSIs)-diflubenzuron, hexaflumuron, or chlorfluazuron-on termite populations. Diflubenzuron at 0.1% active ingredient (AI, wt:wt) had no noticeable effect on termite populations. Chlorfluazuron (0.25% [AI]) significantly reduced termite populations in approximately 3 yr. Chlorfluazuron used after > 2-yr diflubenzuron treatment significantly reduced termite populations within months. This suggests diflubenzuron exposure increased the termite's sensitivity to chlorfluazuron accelerating population collapse. Hexaflumuron (0.5% [AI]) also reduced termite populations in approximately 2 yr. The process of removing most detectable termite populations from the approximately 160,000-m2 campus of the Southern Regional Research Center, New Orleans, LA, with CSIs baits required approximately 3 yr. Adjustments in the specific bait formulations and application procedures might reduce time to suppression. Establishment of new independent termite populations provides a mechanism to minimize the effects of baits. Remedial control measures around and under structures should be considered when implementing an area wide management strategy.

Study of the sustained speed of kill of the combination of fipronil/amitraz/(S)-methoprene and the combination of imidacloprid/permethrin against Dermacentor reticulatus, the European dog tick.

The sustained speed of kill against Dermacentor reticulatus of two topical combinations, one containing fipronil/amitraz/(S)-methoprene and the other, imidacloprid/permethrin, was evaluated in dogs. Two treated groups and one untreated control group of eight adult Beagle dogs each were randomly formed based on pre-infestation rates and bodyweight. Each treatment was administered topically once on Day 0, according to the recommended label dose and instructions for use. All dogs were infested with 50 adult unfed D. reticulatus starting on Day 1, then weekly, for a total of five weeks. While most studies determine tick efficacy at 48 hours (h), in this study, all remaining ticks were counted and categorized 24 h following each infestation. The numbers of ticks (living or dead) that remained attached on treated dogs were compared to those on the control animals. The percent reduction of attached ticks (disruption of attachment) at 24 h on dogs treated with fipronil/amitraz/(S)-methoprene remained above 92% for four weeks. The reduction of attached ticks at 24 h on dogs treated with imidacloprid/permethrin did not reach 80% during the entire study. The number of ticks attached at 24 h was significantly (p<0.05) lower in the fipronil/amitraz/(S)-methoprene group than in the imidacloprid/permethrin group in assessments on Days 2, 15, 22, 29 and 36. When assessing efficacy based upon live ticks on treated versus control dogs, fipronil/amitraz/(S)-methoprene 24 h efficacy was above 95% for four weeks, decreasing to 77.8% at Day 36. The 24 h efficacy of imidacloprid/permethrin ranged from 56.2% to 86.7% through Day 29, never achieving 90% throughout the study. The 24-hour efficacy of fipronil/amitraz/(S)-methoprene was significantly (p<0.05) higher than imidacloprid/permethrin at all time points, including Day 36.

Comparative efficacy on dogs of a single topical treatment with the pioneer fipronil/(S)-methoprene and an oral treatment with spinosad against Ctenocephalides felis.

In the study reported here, the pioneer fipronil/(S)-methoprene topical product (Frontline® Plus, Merial Limited, Duluth, GA) was compared to the oral spinosad product (Comfortis® Elanco, Greenfield, IN) for efficacy against adult fleas and preventing egg production. The product presentations, doses and labelling were the one applicable in the USA. Using a standard protocol, 200 cat fleas of mixed sex were applied to dogs on Days 1, 7, 14, 21, 28, 35, and 42. Dogs were combed to remove fleas 24 hours post-infestation, the fleas were counted, collected, and then reapplied to each dog following completion of their respective count. At 48 hours post-infestation, comb counts were performed and fleas were removed. No fleas were collected from any dog in the fipronil/(S)-methoprene group at any 24 or 48 hours post-infestation assessment throughout the six weeks study, yielding a preventive efficacy of 100%. For the spinosad treatment, efficacy was 100% at 24 hours and 48 hours through Day 16, and thereafter declined. The results observed in the spinosad-treated dogs were highly variable between animals. At the 24 and 48 hours counts following the Day 21 infestation, only five of eight spinosad-treated dogs (62.5%) were flea-free. Following the Day  28 infestation, spinosad efficacy fell to 85% and 89%, for the 24 hours and 48 hours counts, and only two dogs (25%) were flea free, compared to 100% flea-free dogs in the fipronil/(S)-methoprene group. No fleas were collected from the fipronil/(S)-methoprene treated dogs throughout the entire study, therefore, no eggs were collected at any time from any dog in the group. However, in the spinosad group adult fleas were found on dogs starting on Day 21 and by Day 30, 42 eggs were collected from one dog that had 107 adult fleas counted at 48 hours. At Day 37 and Day 49, more than 100 eggs were collected from each dog in the spinosad-treated and control groups.

Field-based evaluation of novaluron EC10 insect growth regulator, a chitin synthesis inhibitor against dengue vector breeding in leaf axils of pineapple plantations in Gampaha District, Sri Lanka.

BACKGROUND: Insect growth regulators (IGRs) are considered a novel group of insecticides to control mosquitoes. Novaluron is an IGR with benzoylphenyl urea insecticide, which inhibits chitin synthesis in insects and can reduce insect population density; it is also known to have a high margin of safety for mammals. METHODS: The effective minimum concentration of novaluron formulation EC10 was tested. Six pineapple plantations [control (n = 3) and test (n = 3)] were selected from Meerigama Medical Officer of Health area in Gampaha District, Sri Lanka. Fifteen plots (10 × 10 m) were demarcated in each site with a 200 m distance apart. Leaf axils of 450 pineapple plants (30 plants × 15 plots) were screened for immature stages of Aedes mosquitoes weekly for 12 weeks. The required concentration (20 ppm) of novaluron was sprayed onto the selected pineapple plants (n = 1350) individually in 3 selected test sites for 5-10 s. The reduction in the vector population was interpreted as the percentage of reduction in immature stages of Aedes mosquitoes. RESULTS: The 100% mortality of the Ae. aegypti larvae within 24 h was observed at 20 ppm (0.05 ml of novaluron 100 g/l in 250 ml of water) as the minimum dose. Variation in the number of Aedes larvae present in the control and intervention sites was found to be significantly different throughout the entire observational period (χ2 = 128.29, df = 11, P < 0.001). The total elimination of Aedes larvae continued for up to 2 weeks and a 50% reduction was observed until the 8th week. CONCLUSIONS: The present study emphasizes that novaluron (10% EC) can be used as an effective larvicide at the treatment dose of 20 ppm. The residual effect of the IGR lasted for 12 weeks with a functional efficacy of 8 weeks. The 100% reduction of larval breeding was observed up to the 2nd week after application and the percentage reduction of immature stages remained > 50% until the 8th week. The lowest reduction (34.2%) was observed at 12 weeks after the initial treatment. Therefore, re-treatment may be recommended based on the reduction in the efficacy of the IGR.

Efficacy of methoprene applied at different temperatures and rates on surface substrates to control eggs and fifth instars of Plodia interpunctella.

A series of studies was conducted to determine the effects of temperature on toxicity of the insect growth regulator methoprene to eggs and larvae of Plodia interpunctella (Hübner), the Indianmeal moth. When methoprene was applied to Kraft paper at the rate of 0.0003 mg of active ingredient [(AI)]/cm2, there was little direct toxicity against eggs of P. interpunctella, and temperature did not affect insecticide efficacy. Similarly, exposure of eggs on a paperboard surface treated with different rates of methoprene resulted in delayed adult emergence but not a reduction in adult emergence. However, wandering-phase larvae ofP. interpuctella were susceptible to methoprene, and exposure of larvae for 0.5, 1, and 2 h on different packaging materials resulted in reduced adult emergence. There was variation in emergence depending on the specific surface, but temperature had no effect on resulting adult emergence from exposed larvae. A partial budget analysis described treatment costs and reduction of risks associated with control of eggs and larvae of P. interpunctella. Results indicate methoprene could be used in management programs to control larvae of P. interpunctella, but eggs may be able to compensate for exposure to methoprene residues on treated surfaces.

Strategic use of crutching and dicyclanil to protect unmulesed sheep against breech strike.

OBJECTIVE: To test strategies for the application of dicyclanil and mid-season crutching to maximise protection of unmulesed sheep against breech strike. PROCEDURE: Three hundred and eighty unmulesed Merino weaners were randomly allocated to four groups either left untreated or treated by different strategies with 50 g/L dicyclanil. Treatments included breech treatment alone and breech plus body treatment, with two application times, immediately after shearing and 6 weeks after crutching or shearing. To assess protection, larval implants with newly hatched Lucilia cuprina larvae were applied to 10 different sheep from each group at 3, 4, 5 and 6 months after crutching and shearing and assessed for the development of strike at 48 hours. The concentration of dicyclanil was measured in wool samples clipped from the breeches of the test sheep. RESULTS: All dicyclanil treatments gave significant reduction in strike in comparison to controls up until 4 months after crutching but protection in the sheep treated immediately after shearing had waned at 5 months. Treating at 6 weeks after crutching provided significant reduction (P < 0.05) in strike for 6 months. Results for strike incidence immediately after shearing and concentration of dicyclanil in the breech wool also suggested improvements in protection by delaying treatment for 6 weeks. CONCLUSION: In most environments it should be possible to protect unmulesed sheep against breech strike with a carefully planned integrated control program incorporating strategically timed crutching, shearing and dicyclanil application. Delaying treatment with dicyclanil to at least 6 weeks after shearing or crutching increased the protection provided in comparison to treatment immediately after shearing.

Burkholderia gut symbiont modulates titer of specific juvenile hormone in the bean bug Riptortus pedestris.

Recent studies have provided molecular evidence that gut symbiotic bacteria modulate host insect development, fitness and reproduction. However, the molecular mechanisms through which gut symbionts regulate these aspects of host physiology remain elusive. To address these questions, we prepared two different Riptortus-Burkholderia insect models, Burkholderia gut symbiont-colonized (Sym) Riptortus pedestris insects and gut symbiont-noncolonized (Apo) insects. Upon LC-MS analyses, juvenile hormone III skipped bisepoxide (JHSB3) was newly identified from Riptortus Apo- and Sym-female and male adults' insect hemolymph and JHSB3 titer in the Apo- and Sym-female insects were measured because JH is important for regulating reproduction in adult insects. The JHSB3 titer in the Sym-females were consistently higher compared to those of Apo-females. Since previous studies reported that Riptortus hexamerin-α and vitellogenin proteins were upregulated by the topical abdominal application of a JH-analog, chemically synthesized JHSB3 was administered to Apo-females. As expected, the hexamerin-α and vitellogenin proteins were dramatically increased in the hemolymph of JHSB3-treated Apo-females, resulting in increased egg production compared to that in Sym-females. Taken together, these results demonstrate that colonization of Burkholderia gut symbiont in the host insect stimulates biosynthesis of the heteroptera-specific JHSB3, leading to larger number of eggs produced and enhanced fitness in Riptortus host insects.