Methoprene, a juvenile hormone analog, is used to accelerate sexual maturation in males of species of economic importance in support to the sterile insect technique (SIT). In the SIT, mass-reared sterile males are released into the field and need to survive until they reach sexual maturation, find a wild female, mate with her and then induce female sexual refractoriness, so she will not remate with a wild counterpart. The use of methoprene shortens the time between release and copulation. However, in South American fruit flies, Anastrepha fraterculus, the ability of methoprene-treated males to inhibit female remating has been shown to be lower than wild males, when methoprene was applied by pupal immersion or topical application. Here we evaluated the possibility of incorporating methoprene into the male diet at different doses and the ability of those males to inhibit female remating, as well as the effect of methoprene on male reproductive organ size, due to the possible correlation between male accessory gland size and their content, and the role of male accessory gland proteins in female inhibition. We found that A. fraterculus males fed with methoprene in the adult protein diet at doses as high as 1% were less likely to inhibit female remating, however, at all other lower doses males had the same ability as untreated males to inhibit female remating. Males fed with methoprene had bigger male accessory glands and testes compared to methoprene-deprived males. We demonstrate that the incorporation of methoprene in adult male diets is possible in this species and potentially useful as a post-teneral, pre-release supplement at doses as low as 0.01%. Even at higher doses, the percentage of females remating after 48 h from the first copulation is sufficiently low in this species so as not compromise the efficiency of the SIT.

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.

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.

The effects of sublethal concentrations of the insect growth regulators (IGRs) diflubenzuron and methoprene on some fitness components of an Aedes aegypti (L.) population in south-eastern Brazil were investigated. The fourth instar larvae of the mosquito were exposed to 20 ppb methoprene and 3 ppb diflubenzuron, and adult survivors were evaluated for fitness parameters, including body size and biomass, fecundity, fertility, longevity and symmetry. The sublethal concentrations had negative effects on longevity, but biomass, fecundity and fertility were not affected by either IGR. In separate experiments, A. aegypti individuals' surviving concentrations of 2, 3 and 3.5 ppb diflubenzuron and 5, 10 and 20 ppb methoprene were used for morphological analyses (size and symmetry). Diflubenzuron had negative effects on the mosquito's body symmetry. The observed decreases in longevity and symmetry may have negative impacts on the population dynamics of A. aegypti.

Total and differential haemocyte counts following starvation and feeding resumption, ventral nerve cord transection, and brain implantation followed by methoprene application and 20-hydroxyecdysone injection were studied in Danaus chrysippus (Linnaeus) larvae. The results revealed that initial starvation and transection of the ventral nerve cord before the fifth abdominal ganglion in fifth instars of D. chrysippus led to a reduced total haemocyte count, on the one hand, and to a higher variation in percentage of different haemocyte types, on the other. While feeding resumption, brain implantation and 20-hydroxyecdysone treatments reversed the effect of starvation and nerve cord transection, methoprene application produced a decline in total number of haemolymph cells.

The juvenoids fenoxycarb and methoprene were evaluated for morphogenetic activity on last-instar nymphs of the American bird grasshopper, Schistocerca americana (Drury) (Orthoptera: Acrididae). Topical application of fenoxycarb induced supernumerary nymphs, adultoids and adults with curled wings. Methoprene yielded similar morphogenetic effects although at reduced levels of activity. Of particular interest, however, is the discovery that, in addition to its morphogenetic activity, fenoxycarb causes significant mortality in developing fourth-, fifth- and sixth-instar nymphs.

The leaf surface treatment given to nymphal instars of Lipaphis erysimi with methoprene (1–10,000 ppm) prolonged the developmental period only in the second instar. The percentage emergence decreased significantly in the treatments of first and second instar and the alate production increased in the treatment of the latter instar only. In addition, dose-dependent morphological abnormalities were also observed. The longevity and fecundity were affected adversely when adults were given both topical and leaf surface treatments. However, these effects were not transmitted to the F1 generation.

Seven benzyl-1,3-benzodioxole analogs were tested for possible effects on reproduction in females of the Mediterranean fruit fly. Single topical doses of J2581 (5-ethoxy-6-[4-methoxyphenyl]methyl-1,3-benzodioxole) and J3263 (5-methoxy-6-(1-[4-ethoxyphenyl]ethyl)-1,3-benzodioxole) were found to cause a temporary delay in ovarian growth in female medflies by 9 and 13 days respectively when compared to acetone-treated controls. Continued exposure of the analogs at the target site(s) appears to be necessary for more permanent effects. Nurse cell development appears not to be affected, but oocyte growth and subsequent egg production and egg hatch were significantly reduced. Methoprene treatment appeared to obviate the effect of J2581 in arresting ovarian growth.