Juvenile Hormone Mimic Research Articles

Identification of differentially expressed genes in the pheromone glands of mated and virgin Bombyx mori by digital gene expression profiling.

BackgroundMating decreases female receptivity and terminates sex pheromone production in moths. Although significant progress has been made in elucidating the mating-regulated inactivation of pheromone biosynthesis-activating neuropeptide (PBAN) secretion, little is known about the mating induced gene expression profiles in pheromone glands (PGs). In this study, the associated genes involved in Bombyx mori mating were identified through digital gene expression (DGE) profiling and subsequent RNA interference (RNAi) to elucidate the molecular mechanisms underlying the mating-regulated gene expression in PGs.ResultsEight DGE libraries were constructed from the PGs of mated and virgin females: 1 h mating (M1)/virgin (V1) PGs, 3 h mating (M3)/virgin (V3) PGs, 24 h mating (M24)/virgin (V24) PGs and 48 h mating (M48)/virgin (V48) PGs (M48 and V48). These libraries were used to investigate the gene expression profiles affected by mating. DGE profiling revealed a series of genes showing differential expression in each set of mated and virgin female samples, including immune-associated genes, sex pheromone synthesis-associated genes, juvenile hormone (JH) signal-associated genes, etc. Most interestingly, JH signal was found to be activated by mating. Application of the JH mimics, methoprene to the newly-emerged virgin females leaded to the significant reduction of sex pheromone production. RNAi-mediated knockdown of putative JH receptor gene, Methoprene tolerant 1 (Met1), in female pupa resulted in a significant decrease in sex pheromone production in mature females, suggesting the importance of JH in sex pheromone synthesis.ConclusionA series of differentially expressed genes in PGs in response to mating was identified. This study improves our understanding of the role of JH signaling on the mating-elicited termination of sex pheromone production.

Interactive Effects of Mosquito Control Insecticide Toxicity, Hypoxia, and Increased Carbon Dioxide on Larval and Juvenile Eastern Oysters and Hard Clams

Mosquito control insecticide use in the coastal zone coincides with the habitat and mariculture operations of commercially and ecologically important shellfish species. Few data are available regarding insecticide toxicity to shellfish early life stages, and potential interactions with abiotic stressors, such as low oxygen and increased CO2 (low pH), are less understood. Toxicity was assessed at 4 and 21days for larval and juvenile stages of the Eastern oyster, Crassostrea virginica, and the hard clam, Mercenaria mercenaria, using two pyrethroids (resmethrin and permethrin), an organophosphate (naled), and a juvenile growth hormone mimic (methoprene). Acute toxicity (4-day LC50) values ranged from 1.59 to >10mg/L. Overall, clams were more susceptible to mosquito control insecticides than oysters. Naled was the most toxic compound in oyster larvae, whereas resmethrin was the most toxic compound in clam larvae. Mortality for both species generally increased with chronic insecticide exposure (21-day LC50 values ranged from 0.60 to 9.49mg/L). Insecticide exposure also caused sublethal effects, including decreased swimming activity after 4days in larval oysters (4-day EC50 values of 0.60 to 2.33mg/L) and decreased growth (shell area and weight) in juvenile clams and oysters after 21days (detected at concentrations ranging from 0.625 to 10mg/L). Hypoxia, hypercapnia, and a combination of hypoxia and hypercapnia caused mortality in larval clams and increased resmethrin toxicity. These data will benefit both shellfish mariculture operations and environmental resource agencies as they manage the use of mosquito control insecticides near coastal ecosystems.

Toxicological and Biological Effects of Juvenile Hormone Mimic (JHM) Pyriproxyfen Against the Two Spotted Spider Mite, Tetranychus urticae Koch (Acari: Tetranychidae)

The present investigation was carried out to study the effect of Juvenile Hormone Mimic (JHM), Pyriproxyfen 10% EC on toxicological and biological aspects of the two spotted spider mite, Tetranychus urticae Koch under laboratory conditions. The effects of Pyriproxyfen on mortality, life cycle, duration period, adult longevity, ovipositional period and female fecundity were evaluated. Results indicated that the candidate compound exhibited direct contact toxicity against all the different developmental stages of T. urticae, but protonymphs were more susceptible than adult females. The LC50's were 4.21 and 4.36 ppm after 72 hr. post exposure for protonymphs and females, respectively. Also it had contact ovicidal activity influenced by egg age. 48-72 hr. eggs were significantly more sensitive than eggs of the other ages. According to LC50 values LC50's were 117.07, 7.95 and 2.91ppm for the three tested ages (0-24 hr, 24-48 hr and 48-72 hr old), respectively. Also, all the tested concentrations prolongated the incubation, larval, protonymphal and deutonymphal periods, but the maximum conc. level (10 ppm) produced the highest increase. Also, the longevity of males and females were increased, the highest increase was observed with 10 ppm, it produced 23.42 days/♀ and 18.5 days/♂ compared with 15.59 days/♀ and 15.92 days/♂ in the control. On the otherhand, oviposition periods were also influenced as concentration dependant response. The mean number of laid eggs/♀ were highly decreased as conc. increased, the highest decrease was observed with T. urticae females treated with conc. 10 ppm, it gave 14.17 eggs/♀ compared with 124 eggs/♀ in untreated females. Results suggest that JHM, pyriproxyfen could be a good alternative and interesting component in Integrated Pest Management (IPM) for controlling TSSM, T. urticae and minimizing the usage of conventional pesticides.

Juvenile hormone signaling during reproduction and development of the linden bug, Pyrrhocoris apterus

Juvenile hormone (JH), a sesquiterpenoid produced by the insect corpus allatum gland (CA), prevents metamorphosis in larvae and stimulates vitellogenesis in adult females. Whether the same JH signaling pathway regulates both processes is presently unknown. Here, we employ the robust JH response during reproduction and development of the linden bug, Pyrrhocoris apterus, to compare the function of key JH-signaling genes encoding the JH receptor, Methoprene-tolerant (Met), its binding partner Taiman (Tai), and a JH-inducible protein, Krüppel-homolog 1 (Kr-h1). RNA interference (RNAi) with Met or Tai, but not Kr-h1, blocked ovarian development and suppressed vitellogenin gene expression in the fat body of females raised under reproduction-inducing conditions. Loss of Met and Tai matched the effects of CA ablation or the natural absence of JH during reproductive diapause. Stimulation of vitellogenesis by treatment of diapausing females with a JH mimic methoprene also required both Met and Tai in the fat body, whereas Kr-h1 RNAi had no effect. Therefore, the Met-Tai complex likely functions as a JH receptor during vitellogenesis. In contrast to Met and Kr-h1 that are both required for JH to prevent precocious metamorphosis in P. apterus larvae, removal of Tai disrupted larval ecdysis without causing premature adult development. Our results show that while Met operates during metamorphosis in larvae and reproduction in adult females, its partner Tai is only required for the latter. The diverse functions of JH thus likely rely on a common receptor whose actions are modulated by distinct components.

Pyriproxyfen enhances the immunoglobulin G immune response in mice

Pyriproxyfen is a juvenile hormone mimic of vital importance for insect development with little risk to humans. This study was performed to investigate whether large doses of pyriproxyfen affect the immune response in mammals. Mice were immunized thrice with ovalbumin in 5% ethanol, with or without pyriproxyfen or alum. Large doses of pyriproxyfen (9 or 15 mM) significantly enhanced specific total IgG immune response. This enhancement was no longer present 24 hr after treatment with pyriproxyfen. These results suggest that pyriproxyfen is a safe chemical. Moreover, pyriproxyfen induced higher titers of IgG2a and enhanced tumor necrosis factor-alpha and gamma-interferon responses whereas alum induced IgG1 with enhanced interleukin-4 and -10. These observations indicate that the mechanism of immune enhancement by pyriproxyfen may differ from that of alum.

Remating behavior in Anastrepha fraterculus (Diptera: Tephritidae) females is affected by male juvenile hormone analog treatment but not by male sterilization

The sterile insect technique (SIT) has been proposed as an area-wide method to control the South American fruit fly, Anastrepha fraterculus (Wiedemann). This technique requires sterilization, a procedure that affects, along with other factors, the ability of males to modulate female sexual receptivity after copulation. Numerous pre-release treatments have been proposed to counteract the detrimental effects of irradiation, rearing and handling and increase SIT effectiveness. These include treating newly emerged males with a juvenile hormone mimic (methoprene) or supplying protein to the male's diet to accelerate sexual maturation prior to release. Here, we examine how male irradiation, methoprene treatment and protein intake affect remating behavior and the amount of sperm stored in inseminated females. In field cage experiments, we found that irradiated laboratory males were equally able to modulate female remating behavior as fertile wild males. However, females mated with 6-day-old, methoprene-treated males remated more and sooner than females mated with naturally matured males, either sterile or wild. Protein intake by males was not sufficient to overcome reduced ability of methoprene-treated males to induce refractory periods in females as lengthy as those induced by wild and naturally matured males. The amount of sperm stored by females was not affected by male irradiation, methoprene treatment or protein intake. This finding revealed that factors in addition to sperm volume intervene in regulating female receptivity after copulation. Implications for SIT are discussed.

Designing and Binding Mode Prediction of Juvenile Hormone Analogues as Potential Inhibitor for Galleria mellonella

Virtual screening of chemical databases has become an integral part of ligand design. Docking is one of the most important methods in computer assisted screening. If a three dimensional structure of target receptor is available, along with information regarding nature of the ligand-binding site, ligand-binding mode; the interactions between the ligand and receptor can be studied extensively, in order to design and develop target specific new compounds in a short time period. Juvenile Hormone Analogues, sesqui-terpenoid series of compound act as an insect growth regulator, and presently in use as a potential environment friendly pesticide. Juvenile hormone is the molting hormone responsible for each molt, and involved in a wide range of physiological processes in both developing and mature insect. Designing of various juvenile hormone analogues are new and emerging area to counter the insect problem. In this paper, we report protein-ligand interactions using a standard protocol of docking. We perform screening of synthesized (A-B) and proposed (C-D) series of Juvenile Hormone Analogues with hemolymph binding proteins of Galleria mellonella. Further binding energy profile of all the series have been compared with the phenoxy derivatives of juvenile hormone mimics, as well as natural JH III, in order to design targeted JHAs with improved biological activities. Our proposed series of juvenile hormone analogues exhibit better energy profile over in use phenoxy derivatives.

Preliminary studies developing methods for the control of Chrysomya putoria, the African latrine fly, in pit latrines in The Gambia

ObjectiveTo explore ways of controlling Chrysomya putoria, the African latrine fly, in pit latrines. As pit latrines are a major source of these flies, eliminating these important breeding sites is likely to reduce village fly populations, and may reduce the spread of diarrhoeal pathogens.MethodsWe treated 24 latrines in a Gambian village: six each with (i) pyriproxyfen, an insect juvenile hormone mimic formulated as Sumilarv® 0.5G, a 0.5% pyriproxyfen granule, (ii) expanded polystyrene beads (EPB), (iii) local soap or (iv) no treatment as controls. Flies were collected using exit traps placed over the drop holes, weekly for five weeks. In a separate study, we tested whether latrines also function as efficient flytraps using the faecal odours as attractants. We constructed six pit latrines each with a built-in flytrap and tested their catching efficiency compared to six fish-baited box traps positioned 10 m from the latrine. Focus group discussions conducted afterwards assessed the acceptability of the flytrap latrines.ResultsNumbers of emerging C. putoria were reduced by 96.0% (95% CIs: 94.5–97.2%) 4–5 weeks after treatment with pyriproxyfen; by 64.2% (95% CIs: 51.8–73.5%) after treatment with local soap; by 41.3% (95% CIs = 24.0–54.7%) after treatment with EPB 3–5 weeks after treatment. Flytraps placed on latrines collected C. putoria and were deemed acceptable to local communities.ConclusionsSumilarv 0.5G shows promise as a chemical control agent, whilst odour-baited latrine traps may prove a useful method of non-chemical fly control. Both methods warrant further development to reduce fly production from pit latrines. A combination of interventions may prove effective for the control of latrine flies and the diseases they transmit.

Precocious sexual signalling and mating in Anastrepha fraterculus (Diptera: Tephritidae) sterile males achieved through juvenile hormone treatment and protein supplements

Sexual maturation of Anastrepha fraterculus is a long process. Methoprene (a mimic of juvenile hormone) considerably reduces the time for sexual maturation in males. However, in other Anastrepha species, this effect depends on protein intake at the adult stage. Here, we evaluated the mating competitiveness of sterile laboratory males and females that were treated with methoprene (either the pupal or adult stage) and were kept under different regimes of adult food, which varied in the protein source and the sugar:protein ratio. Experiments were carried out under semi-natural conditions, where laboratory flies competed over copulations with sexually mature wild flies. Sterile, methoprene-treated males that reached sexual maturity earlier (six days old), displayed the same lekking behaviour, attractiveness to females and mating competitiveness as mature wild males. This effect depended on protein intake. Diets containing sugar and hydrolyzed yeast allowed sterile males to compete with wild males (even at a low concentration of protein), while brewer´s yeast failed to do so even at a higher concentration. Sugar only fed males were unable to achieve significant numbers of copulations. Methoprene did not increase the readiness to mate of six-day-old sterile females. Long pre-copulatory periods create an additional cost to the management of fruit fly pests through the sterile insect technique (SIT). Our findings suggest that methoprene treatment will increase SIT effectiveness against A. fraterculus when coupled with a diet fortified with protein. Additionally, methoprene acts as a physiological sexing method, allowing the release of mature males and immature females and hence increasing SIT efficiency.

Toxicology of insecticides to mammals

Many insecticides target structures or functions in non-target species, including mammals. This is particularly true of those that target the insect nervous system, such as the organochlorines, anticholinesterases and GABA antagonists. Another group of insecticides target structures or functions not present in mammals, and this group of insecticides has considerable target species specificity, but there are often potential targets in mammals. Octopamine is closely related to adrenaline and amitraz (an octopamine receptor agonist) and acts in mammals at α2-adrenergic receptors. Although there are potential targets in mammals for juvenile hormone mimics and ecdysone receptor agonists, there is no evidence that the mammalian toxicity of either group is related to their insecticidal activity. Nor do chitin synthesis inhibitors have high mammalian toxicity.

Over-Expression of a Cytochrome P450 Is Associated with Resistance to Pyriproxyfen in the Greenhouse Whitefly Trialeurodes vaporariorum

BackgroundThe juvenile hormone mimic, pyriproxyfen is a suppressor of insect embryogenesis and development, and is effective at controlling pests such as the greenhouse whitefly Trialeurodes vaporariorum (Westwood) which are resistant to other chemical classes of insecticides. Although there are reports of insects evolving resistance to pyriproxyfen, the underlying resistance mechanism(s) are poorly understood.ResultsBioassays against eggs of a German (TV8) population of T. vaporariorum revealed a moderate level (21-fold) of resistance to pyriproxyfen. This is the first time that pyriproxyfen resistance has been confirmed in this species. Sequential selection of TV8 rapidly generated a strain (TV8pyrsel) displaying a much higher resistance ratio (>4000-fold). The enzyme inhibitor piperonyl butoxide (PBO) suppressed this increased resistance, indicating that it was primarily mediated via metabolic detoxification. Microarray analysis identified a number of significantly over-expressed genes in TV8pyrsel as candidates for a role in resistance including cytochrome-P450 dependent monooxygenases (P450s). Quantitative PCR highlighted a single P450 gene (CYP4G61) that was highly over-expressed (81.7-fold) in TV8pyrsel.ConclusionOver-expression of a single cytochrome P450 gene (CYP4G61) has emerged as a strong candidate for causing the enhanced resistance phenotype. Further work is needed to confirm the role of the encoded P450 enzyme CYP4G61 in detoxifying pyriproxyfen.

Egg exposure to pyriproxyfen in the tomato leaf miner Tuta absoluta: ovicidal activity or behavioural‐modulated hatching mortality?

AbstractInsecticide susceptibility varies with the insect life stage and although the egg stage is sometimes perceived as the most vulnerable, it is a difficult target for insecticide application and little studied as such. Egg susceptibility to insecticides is generally considered important for insect growth regulators and insect pests of reduced mobility like leaf miners because of their placement on the host plant part targeted by the insecticidal application. The egg exposure to the juvenile hormone mimic pyriproxyfen was studied here in the tomato leaf miner Tuta absoluta (Lepidoptera: Gelechiidae), a key tomato insect pest, aiming to assess if mortality is achieved due to insecticide ovicidal activity or due to behavioural impairment of the larvae preventing egg hatching. Survival analysis of treated tomato leaves and/or eggs indicated the importance of egg exposure to pyriproxyfen leading to a significant decrease in survival with a nearly 50% reduction in survival time, which peaked at the first instar. Ovicidal activity of pyriproxyfen in the tomato leaf miner was negligible, but image recording and behaviour analysis indicated behavioural impairment of larval activity compromising hatching and leaf mining by early instar larvae leading to high mortality at this stage (ca 50%). The likely causes and management implications of such findings are discussed.

Common and Distinct Roles of Juvenile Hormone Signaling Genes in Metamorphosis of Holometabolous and Hemimetabolous Insects

Insect larvae metamorphose to winged and reproductive adults either directly (hemimetaboly) or through an intermediary pupal stage (holometaboly). In either case juvenile hormone (JH) prevents metamorphosis until a larva has attained an appropriate phase of development. In holometabolous insects, JH acts through its putative receptor Methoprene-tolerant (Met) to regulate Krüppel-homolog 1 (Kr-h1) and Broad-Complex (BR-C) genes. While Met and Kr-h1 prevent precocious metamorphosis in pre-final larval instars, BR-C specifies the pupal stage. How JH signaling operates in hemimetabolous insects is poorly understood. Here, we compare the function of Met, Kr-h1 and BR-C genes in the two types of insects. Using systemic RNAi in the hemimetabolous true bug, Pyrrhocoris apterus, we show that Met conveys the JH signal to prevent premature metamorphosis by maintaining high expression of Kr-h1. Knockdown of either Met or Kr-h1 (but not of BR-C) in penultimate-instar Pyrrhocoris larvae causes precocious development of adult color pattern, wings and genitalia. A natural fall of Kr-h1 expression in the last larval instar normally permits adult development, and treatment with an exogenous JH mimic methoprene at this time requires both Met and Kr-h1 to block the adult program and induce an extra larval instar. Met and Kr-h1 therefore serve as JH-dependent repressors of deleterious precocious metamorphic changes in both hemimetabolous and holometabolous juveniles, whereas BR-C has been recruited for a new role in specifying the holometabolous pupa. These results show that despite considerable evolutionary distance, insects with diverse developmental strategies employ a common-core JH signaling pathway to commit to adult morphogenesis.

Reproductive tradeoffs of learning in a butterfly

The evolution of learning has long been hypothesized to be limited by fitness trade-offs such as delays in reproduction. We explored the relationship between host learning and reproduction in the cabbage white butterfly, Pieris rapae. The cabbage white female is innately biased to search for common green hosts but can learn to search for rare red hosts. Host learning was shown previously to vary among full-sibling families and to incur costs in terms of host search efficiency and brain size. In the present study, we show that butterflies from full-sib families with relatively better learning performance on red hosts tend to emerge as adults with relatively fewer and less-developed eggs. We also used methoprene, a juvenile hormone mimic, to advance reproduction in female cabbage whites. We found that methoprene-treated butterflies improved host-finding ability less with experience, relative to controls, providing independent evidence of a link between learning and timing of reproduction. Finally, we show that the learning experience itself is associated with additional decreases in lifetime fecundity. These results support a range of theoretical and comparative studies highlighting the importance of fitness tradeoffs in the evolution of learning and cognition.

Evaluation of nontarget effects of methoprene applied to catch basins for mosquito control

The mosquito larvicide methoprene is a juvenile growth hormone mimic that is widely used to control mosquito larvae in stormwater catch basins. This study addresses two concerns pertaining to methoprene's use for mosquito control. First, measurements of methoprene concentrations were made from water in catch basins that had been treated with methoprene and from an adjoining salt pond near where the treated catch basins emptied. The concentrations of methoprene in catch basins and at drainage outlets after application at the rates currently used for mosquito control in southern Rhode Island were 0.5 ppb and lower, orders of magnitude below what has been determined as detrimental to organisms other than mosquitoes. Second, the effects of methoprene on the communities that live in catch basins were evaluated both in simulated catch basins in the laboratory and in actual catch basins in the field. We found no evidence of declines in abundances of any taxa attributable to the application. Furthermore, we found no consistent changes in community-level parameters (e.g., taxonomic richness, and dominance-diversity relationships) related to methoprene application in either field or laboratory trials.

Developmental profile and hormonal regulation of the transcription factors broad and Krüppel homolog 1 in hemimetabolous thrips

In holometabolous insects, Krüppel homolog 1 ( Kr-h1) and broad ( br) are key players in the juvenile hormone (JH) regulation of metamorphosis: Kr-h1 is an early JH-response gene, while br is a transcription factor that directs pupal development. Thrips (Thysanoptera) are classified as hemimetabolous insects that develop directly from nymph to adult, but they have quiescent and non-feeding stages called propupa and pupa. We analyzed the developmental profiles of br and Kr-h1 in the western flower thrips Frankliniella occidentalis (Thripidae) that has one propupal instar and one pupal instar, and Haplothrips brevitubus (Phlaeothripidae) that has one propupal instar and two pupal instars, i.e. pupa I and pupa II. In F. occidentalis, the br mRNA levels were moderate in the embryonic stage, high at the larva-propupa transition, and low in the pre-final larval instar and the pupal stage, while Kr-h1 mRNA levels were high in the embryonic stage, remained at a moderate level in the larval and propupal stages, and low in the pupal stage. The expression profiles in H. brevitubus were very similar to those in F. occidentalis, except that the increase of br expression in the final larval stage occurs more slowly in H. brevitubus, and that the mRNA levels of br and Kr-h1 remained high in pupa I of H. brevitubus and then decreased. These profiles of br and Kr-h1 were comparable to those in holometabolous insects, although br expression found in thrips’ embryogenesis is reminiscent of several hemimetabolous species. Treatment with an exogenous JH mimic (JHM) in distinct developmental stages consistently resulted in lethality as pupa of F. occidentalis or pupa II of H. brevitubus. Treatment with JHM to newly molted propupae caused prolonged expression of Kr-h1 and br in both species, suggesting that Kr-h1 and br could be involved in mediating anti-metamorphic signals of JHM.

A role for juvenile hormone in the prepupal development of Drosophila melanogaster

To elucidate the role of juvenile hormone (JH) in metamorphosis of Drosophila melanogaster, the corpora allata cells, which produce JH, were killed using the cell death gene grim. These allatectomized (CAX) larvae were smaller at pupariation and died at head eversion. They showed premature ecdysone receptor B1 (EcR-B1) in the photoreceptors and in the optic lobe, downregulation of proliferation in the optic lobe, and separation of R7 from R8 in the medulla during the prepupal period. All of these effects of allatectomy were reversed by feeding third instar larvae on a diet containing the JH mimic (JHM) pyriproxifen or by application of JH III or JHM at the onset of wandering. Eye and optic lobe development in the Methoprene-tolerant (Met)-null mutant mimicked that of CAX prepupae, but the mutant formed viable adults, which had marked abnormalities in the organization of their optic lobe neuropils. Feeding Met(27) larvae on the JHM diet did not rescue the premature EcR-B1 expression or the downregulation of proliferation but did partially rescue the premature separation of R7, suggesting that other pathways besides Met might be involved in mediating the response to JH. Selective expression of Met RNAi in the photoreceptors caused their premature expression of EcR-B1 and the separation of R7 and R8, but driving Met RNAi in lamina neurons led only to the precocious appearance of EcR-B1 in the lamina. Thus, the lack of JH and its receptor Met causes a heterochronic shift in the development of the visual system that is likely to result from some cells 'misinterpreting' the ecdysteroid peaks that drive metamorphosis.

Effect of pyriproxyfen, a juvenile hormone mimic, on egg hatch, nymph development, adult emergence and reproduction of the Asian citrus psyllid, Diaphorina citri Kuwayama

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is a vector of bacteria presumably responsible for huanglongbing (HLB) disease in citrus. In this laboratory study, an investigation was made of the activity of pyriproxyfen, a juvenile hormone mimic, on ACP eggs, nymphs and adults to evaluate its potential as a biorational insecticide for inclusion in an integrated pest management (IPM) program for ACP. Irrespective of egg age, timing or method of treatment, a significantly lower percentage of eggs (5-29%) hatched after exposure to 64 and 128 microg mL(-1) of pyriproxyfen. Only 0-36% of early instars (first, second and third) and 25-74% of late instars (fourth and fifth) survived to adults following exposure to 16, 32 and 64 microg mL(-1) of pyriproxyfen. However, 15-20% of adults that emerged following treatment as late instars exhibited morphological abnormalities. Furthermore, pyriproxyfen adversely affected reproduction (fecundity and fertility) of adults that emerged from treated fifth instars or that were treated topically with 0.04 microg as adults. Application of pyriproxyfen at 64 microg mL(-1) resulted in greater inhibition of egg hatch and suppression of adult emergence compared with lower rates. Pyriproxyfen also markedly reduced female fecundity and egg viability for adults that were exposed either as fifth instars or as newly emerged adults. The ovicidal, larvicidal and reproductive effects against ACP suggest that pyriproxyfen is suitable for integration into an IPM program for ACP.

Suppression of JH biosynthesis by JH analog treatment: Mechanism of suppression and roles of allatostatins and nervous connections in the cockroach Diploptera punctata

Juvenile hormone analogs are known to inhibit the production of juvenile hormone (JH) by the corpora allata (CA). However, the mechanism of this inhibition remains undefined. We have used two JH mimics, fenoxycarb and pyriproxyfen, to examine the mechanism of suppression in the cockroach, Diploptera punctata. Denervation experiments demonstrated the importance of nervous connections between the brain and CA for the inhibition of JH biosynthesis by fenoxycarb. Fenoxycarb treatment alters the sensitivity of CA to allatostatin treatment in vitro. Suppression of JH biosynthesis by fenoxycarb following denervation of the CA showed that innervation was in part responsible for the inhibition. Similarly, maximal inhibition by Dippu-AST7 requires intact nervous connections between the brain and CA, particularly during rapid vitellogenesis. qPCR analysis of brain, CA, ovary and midgut extracts revealed that both allatostatin and its receptor Dippu-ASTR2 show increased levels of expression following topical fenoxycarb treatment, particularly in brain tissue on days 4 and 5 of the first gonadotrophic cycle and in CA on day 4. The correlation between inhibition of JH biosynthesis and increased expression of AST and ASTR2 in brains and CA, together with increased sensitivity of CA to allatostatin in vitro, suggests that allatostatin may be one of the effectors by which fenoxycarb inhibits JH biosynthesis.

Enhancing mating performance after juvenile hormone treatment in Anastrepha fraterculus: a differential response in males and females acts as a physiological sexing system

AbstractMethoprene (a mimic of juvenile hormone) treatment can reduce the time required for sexual maturation in Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) males under laboratory conditions, supporting its use as a treatment for sterile males within the context of the sterile insect technique (SIT). We evaluated sexual behaviour, mating competitiveness of methoprene‐treated males, and female readiness to mate after methoprene‐treatment in field cages. The study involved two strains of A. fraterculus from Argentina and Peru, which show several polymorphisms in relation to their sexual behaviour. We also analyzed whether methoprene treatment affected male and/or female behaviour in the same way in these two strains. Methoprene‐treated males were equally competitive with untreated mature males, and became sexually competitive 6 days after emergence (3–4 days earlier than untreated males). In contrast, methoprene did not induce sexual maturation in females or, at least, it did not induce a higher rate of mating in 7‐day‐old females. These results were observed both for the Argentina and the Peru strains. Altogether, our results indicate that methoprene treatment produces sexually competitive males in field cages. In the absence of a genetic sexing system, and when sterile males and females of A. fraterculus are released simultaneously, the fact that females do not respond as do males to the methoprene treatment acts as a physiological sexing effect. Therefore, in the presence of mainly sexually immature sterile females, released sexually mature sterile males would have to disperse in search of wild fertile females, thereby greatly reducing matings among the released sterile insects and thus enhancing sterile insect technique efficiency.