Toxicity and effects of mosquito larvicides methoprene and surface film (Agnique® MMF) on the development and fecundity of the tadpole shrimp Triops newberryi (Packard) (Notostraca: Triopsidae)

We investigated the interactions of tadpole shrimp, a mosquito biological control agent, with the juvenile hormone analog methoprene and a monomolecular surface film. In laboratory assays, the tadpole shrimp (TPS) Triops newberryi (Packard) was able to tolerate high concentrations of methoprene without negative impacts on its growth, longevity, and fecundity when exposed to 1 to 10 mg/liter, or 90–900 fold, of the IE₉₀ levels against a laboratory colony of Culex quinquefasciatus Say. The same held true in field trials when the habitats were treated with Altosid® Liquid Larvicide (Altosid® LL, 5% methoprene) at 0.3–1.2 liters/ha. or 1–4 fold of the label rates for mosquito control. However, some significant impacts on the TPS occurred when they were exposed to Agnique® Monomolecular Film (Agnique® MMF) at the label rates for mosquito control ranging from 1.89–9.45 liters/ha. under laboratory and field conditions. To avoid the negative impact of Agnique MMF on tadpole shrimp, it appears that 1.89 liters/ha. would be the maximum rate when Agnique MMF is used to control mosquitoes in the habitats where the TPS is employed as a biological control agent, or prevailing in the aquatic habitats with potential for suppressing mosquito larval populations.     

Aedes albopictus (Diptera: Culicidae) in Rome: experimental study of relevant control strategy parameters

Since 1997, Aedes albopictus has colonised and then rapidly invaded the city of Rome (Italy) and its peripheral areas. Presently, the control of this mosquito in Italy relies mainly on larvicidal treatment of street storm sewer catch basins with the organophosphate temephos. We have therefore obtained baseline data on the susceptibility to temephos of the Roman Ae. albopictus population by laboratory bioassays on F1 fourth-instar larvae following standard WHO protocols. Estimated lethal concentrations were 0.014 mg/l (LC50) and 0.022 mg/l (LC90) indicating a lack of resistance to this compound. The persistence of temephos in sewer catch basins was evaluated by follow-up of catch basins treated with a dose of 1.5 mg of active ingredient. Mosquito larvae were recovered in 10% and 50% of the treated basins at 9 and 18 days posttreatment, respectively. In order to understand the relative contribution of this larval habitat to adult populations, we conducted a survey in the Zoo of Rome to estimate the larval density of mosquitoes breeding in sewer catch basins. A complete census of a 16.5 ha area mapped 243 catch basins, but only 25 (10.3%) contained water; of the latter 8 (32.0%) hosted mosquito larvae. All positive catch basins contained larvae of Culex pipiens, which were associated with Culiseta longiareolata and/or Ae. albopictus in 6 and 3 cases, respectively. A longitudinal survey of one catch basin over 4 months showed that the mean larval density of Ae. albopictus was markedly lower than that of Cx pipiens and Cs. Iongiareolata, ranging between 0 and 1.3 larvae/dip as compared to 0-33.2 and 0-22.7 larvae/dip, respectively. However, adult densities of Ae. albopictus in this area estimated during the same period with 20 ovitraps showed consistently high values (proportion of positive ovitraps around 100%). These preliminary observations suggest that whenever alternative larval biotopes other than sewer catch basins are widely available, they might be more productive and/or preferred substrates to catch basins for Ae. albopictus breeding.     

Reduced productivity of Culex pipiens and Cx. restuans (Diptera: Culicidae) mosquitoes in parking area catch basins in the northeast Chicago metropolitan area

From June to September, 2016, 100 catch basins in eight parking areas were monitored weekly for the presence of mosquito pupae in the operational area of the North Shore Mosquito Abatement District (NSMAD) located just north of Chicago, IL, U.S.A. Weekly results from these basins were compared to weekly samples taken from residential street catch basins, the most common type of catch basin treated seasonally by the NSMAD with larvicides. Over the 17 study weeks, residential street basins had a mean rate of productivity (pupae per basin‐visit) 12 times that of parking area catch basins. The two parking area sites with the highest mean rate of productivity were associated with county forest preserves. Productivity in both street and parking area basins was positively associated with the presence of three or more deciduous trees within 20 m of basins and if they were located directly adjacent to curbs. Alternatively, productivity was negatively associated with the proportion of impervious surface within 10 m of basins and weekly rainfall. Findings suggest that reduced catch basin larvicide applications may be appropriate in many parking area sites.     

Evaluation of methoprene effect on Aedes aegypti (Diptera: Culicidae) development in laboratory conditions

Several Brazilian Aedes aegypti populations are resistant to the larvicidae temephos. Methoprene, that inhibits adult emergence, is one of the alternatives envisaged by the Brazilian Dengue Control Program (PNCD). However, at Brazil vector infestation rates are measured through larvae indexes and it has been claimed that methoprene use in the field could face operational problems. In order to define a standardized protocol, methoprene effect was evaluated in laboratory conditions after continuous exposure of larvae (Rockefeller strain) to a methoprene formulation available to the PNCD. Methoprene-derived mortality occurs mainly at the pupa stage and pupa development is inversely proportional to methoprene concentration. Number and viability of eggs laid by treated and control females are equivalent. A methoprene dose-dependent delay in the development was noted; however, strong correlations were found for total mortality or adult emergence inhibition if data obtained when all control mosquitoes have emerged are compared to data obtained when methoprene-treated groups finish development. The cumulative record of total methoprene-induced mortality at the time control adults emerge is proposed for routine evaluation of field populations. Mortality of all specimens, but not of larva, could account for adult emergence inhibition, confirming the inadequacy of larvae indexes to evaluate methoprene effect.     

Methoprene does not affect juvenile hormone titers in honey bee (Apis mellifera) workers

Methoprene, a juvenile hormone (JH) analog, is a widely used insecticide that also accelerates behavioral development in honey bees (Apis mellifera). JH regulates the transition from nursing to foraging in adult worker bees, and treatment with JH or methoprene have both been shown to induce precocious foraging. To determine how methoprene changes honey bee behavior, we compared JH titers of methoprene‐treated and untreated bees. Behavioral observations confirmed that methoprene treatment significantly increased the number of precocious foragers in 3 out of 4 colonies. In only 1 out of 4 colonies, however, was there a significant difference in JH titers between the methoprene‐treated and control bees. Further, in all 4 colonies, there was no significant differences in JH titers between precocious and normal‐aged foragers. These results suggest that methoprene did not directly affect the endogenous JH secreted by corpora allata. Because methoprene caused early foraging without changing workers’ JH titers, we conclude that methoprene most likely acts directly on the JH receptors as a substitute for JH.     

Methoprene application and diet protein supplementation to male melon fly,Bactrocera cucurbitae,modifies female remating behavior

Methoprene (an analogue of juvenile hormone) application and feeding on a protein diet is known to enhance male melon fly, Bactrocera cucurbitae Coquillett (Diptera: Tephritidae), mating success. In this study, we investigated the effect of these treatments on male B. cucurbitae's ability to inhibit female remating. While 14‐d‐old females were fed on protein diet, 6‐d‐old males were exposed to one of the following treatments: (i) topical application of methoprene and fed on a protein diet; (ii) no methoprene but fed on a protein diet; (iii) methoprene and sugar‐fed only; and (iv) sugar‐fed, 14‐d‐old males acted as controls. Treatments had no effect on a male's ability to depress the female remating receptivity in comparison to the control. Females mated with protein‐deprived males showed higher remating receptivity than females first mated with protein‐fed males. Methoprene and protein diet interaction had a positive effect on male mating success during the first and second mating of females. Significantly more females first mated with sugar‐fed males remated with protein‐fed males and females first mated with methoprene treated and protein‐fed males were more likely to remate with similarly treated males. Females mating latency (time to start mating) was significantly shorter with protein‐fed males, and mating duration was significantly longer with protein‐fed males compared with protein‐deprived males. These results are discussed in the context of methoprene and/or dietary protein as prerelease treatment of sterile males in area‐wide control of melon fly integrating the sterile insect technique (SIT).     

Evaluation of the triflumuron and the mixture of Bacillus thuringiensis plus Bacillus sphaericus for control of the immature stages of Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae) in catch basins

INTRODUCTION: In Cali, Colombia, catch basins (streetside storm drains) are one of the main larval habitats of Aedes aegypti and Culex quinquefasciatus. Since 1999, these mosquitoes have been controlled by the Secretaría de Salud Municipal (Secretary of Municipal Public Health) using the larvicide triflumuron. Because of high densities of these mosquitoes that remain in the city, treatment failure was suspected -possibly insecticide resistance of the target species. OBJECTIVES: The efficacy of triflumuron and VectoMax (biorational mixture of Bacillus thuringiensis var. israelensis plus Bacillus sphaericus) were evaluated in the control of A. aegypti and C. quinquefasciatus in catch basins. The residual effect of a single application of the biorational formulation was determined in catch basins during periods of high and low rainfall. MATERIALS AND METHODS: The efficacy of the products was measured in 60 catch basins located in a residential neighborhood of Cali for a period of 90 days. The mean number of immature instars (A. aegypti and C. quinquefasciatus larvae and pupae of both species) was determined biweekly from 40 catch basins with insecticide intervention (20 treated with triflumuron, 20 with VectoMax) and 20 untreated (control group). The residual effect of the biorational larvicide was evaluated biweekly in 10 catch basins during each of the 2 climatic periods. Results. The catch basins treated with VectoMax presented a significantly lower mean number of immature instars of both species compared with the control ( p<0.01). In contrast, the triflumuron treatment significantly reduced only immature instars of A. aegypti compared with the control ( p<0.001). The residual effect of VectoMax was higher during low rainfall compared to the control ( p<0.001). Conclusion. The biorational formulation was the more effective treatment for the control of both species during the period of evaluation (15 days).     

Effects of the juvenile hormone analogue methoprene and dietary protein on male melon fly Bactrocera cucurbitae (Diptera: Tephritidae) mating success

The effect of access to dietary protein (P) and the topical application of a juvenile hormone analogue (methoprene (M)) on mating behaviour of male melon fly Bactrocera cucurbitae was assessed in the laboratory and in field cages. Age, dietary protein and methoprene application increased the mating success and influenced the mating behaviour. Treatment with methoprene (M+) to protein-deprived (P−) males had only a modest effect on the acceleration of sexual maturity, but application of methoprene (M+) to protein-fed (P+) males greatly accelerated sexual maturity. Protein diet (P+) increased mating success of males in comparison to protein-deprived (P−) males. Protein and methoprene have a synergistic effect on mating behaviour, since M + P+ treated males exhibit reduced mating latency and achieved higher mating in younger ages than methoprene and/or protein-deprived males. Copulation duration was correlated with nutritional status and M + P+ males copulated longer at the age of advanced sexual maturity than M − P+ males. Our results suggest that in this species with a lek mating system, females discriminate between the males based on their sexual signals, which were influenced by protein in the adult diet, methoprene application and age. The results are discussed in the light of mating competitiveness of precocious treated young males and their relevance to Sterile Insect Technique application against this pest species.     

Effects of nutrition and methoprene treatment upon reproductive diapause in Caloptilia fraxinella (Lepidoptera: Gracillariidae)

Abstract Female Caloptilia fraxinella exhibit a prolonged reproductive diapause immediately post adult emergence in mid‐summer until the next spring when mating, egg development and oviposition on fresh Fraxinus spp. leaflets occur. Factors that effect the termination of reproductive diapause are investigated in this species. Caloptilia fraxinella diapausing adults held in overwintering conditions (2 °C, LD 0 : 24 h) for 24 weeks terminate diapause after placement for 2 weeks in simulated summer conditions (24 °C, LD 16 : 8 h) only if they are provided with 10% sugar water. Exogenous application of the Juvenile Hormone (JH) analogue methoprene to moths in both early‐ (summer) and mid‐ (autumn) reproductive diapause demonstrates that JH affects diapause termination but a carbohydrate nutrition source also mediates mating and vitellogenesis. Mating between moth pairs early in diapause occurs only after treatment with methoprene and provision with sugar water. However, there is no impact of mating on the propensity of females to produce vitellogenic oöcytes. Moths collected in the autumn in mid‐diapause respond in a dose‐dependent fashion to methoprene treatment and the response is greater than that of moths early in diapause collected in the summer. Treatment with methoprene and access to sugar water results in vitellogenic oöcytes in 18.75% of females from mid‐diapause moth pairs treated with 0.01 μg methoprene per insect and in all females from pairs treated at the two highest doses of methoprene (0.1 and 1 μg per insect). Mating occurs only between moths in mid‐diapause treated with the two highest doses of methoprene and these doses induce 91% and 100% mating, respectively. Both control and methoprene‐treated males in mid‐diapause held under summer conditions mate successfully and pass a spermatophore to their methoprene‐treated female partner. These data demonstrate that female C. fraxinella undergo a prolonged reproductive diapause in which termination is dependent on JH and further mediated by a carbohydrate nutrition source. The production of vitellogenic oöcytes is independent of mating. These data also provide evidence that response of moths in diapause to exogenous applications of methoprene differs throughout the diapause period and between male and female C. fraxinella.     

Methoprene modulates the effect of diet on male melon fly,Bactrocera cucurbitae,performance at mating aggregations

The effect of access to dietary protein (P) (hydrolyzed yeast) and/or treatment with a juvenile hormone analogue, methoprene (M), (in addition to sugar and water) on male aggregation (lekking) behaviour and mating success was studied in a laboratory strain of the melon fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae). Six‐day‐old males were treated with (1) protein and methoprene (M+P+), (2) only protein (M?P+), or (3) only methoprene (M+P?), and compared with 14‐day‐old sexually mature untreated males (M?P?). The lekking behaviour of the four groups of males when competing for virgin sexually mature females (14 –16 days old) was observed in field cages. The following parameters were measured at male aggregations: lek initiation, lek participation, males calling, male–male interaction, female acceptance index, and mating success. For all these parameters, the M+P+ males significantly outperformed the other males. Moreover, for all parameters, there was a similar trend with M+P+ > M?P+ > M?P? > M+P?. More M+P+ males called and initiated and participated in lek activities than all other types of male, which resulted in higher mating success. They had also fewer unsuccessful copulation attempts than their counterparts. Whereas treatment with methoprene alone had a negative effect in young males with only access to sugar, access to dietary protein alone significantly improved young male sexual performance; moreover, the provision of methoprene together with protein had a synergistic effect, improving further male performance at leks. The results are of great relevance for enhancing the application of the sterile insect technique (SIT) against this pest species. The fact that access to dietary protein and treatment of sterile males with methoprene improves mating success means that SIT cost‐effectiveness is increased, as more released males survive to sexual maturity.     

The impact of insecticide‐treated cloth targets on the survival of Stegomyia polynesiensis (= Aedes polynesiensis) under laboratory and semi‐field conditions in French Polynesia

The impact of deltamethrin‐impregnated cloth targets on Stegomyia polynesiensis (= Aedes polynesiensis) (Marks) (Diptera: Culicidae) was assessed under laboratory and semi‐field settings in French Polynesia. Stegomyia polynesiensis females were released into small laboratory cages and large field cages containing either a deltamethrin‐treated or an untreated navy blue cloth, and mosquito knock‐down and mortality were assessed. The 24‐h mortality rate in mosquitoes exposed to the insecticide‐treated target in small cages was 98.0%. These mosquitoes also demonstrated significantly higher levels of knock‐down than those exposed to the untreated target. Mortality in field cages was assessed at 24 and 48 h. The 24‐h mortality rate in mosquitoes exposed to the control target was 31.2%, whereas that in those exposed to the deltamethrin‐treated target was 54.3%. The 48‐h mortality rate was also elevated in mosquitoes exposed to the deltamethrin‐treated target, but this result did not differ significantly from that observed in mosquitoes exposed to the control target. The significant suppression of female S. polynesiensis by deltamethrin‐treated resting targets in this study indicates that these targets could play a role in the control of an important disease vector in the South Pacific region.     

Limb regeneration and molting processes under chronic methoprene exposure in the mud fiddler crab, Uca pugnax

Insect growth regulator application for wetland mosquito control remains controversial due to the potential for disruption of normal development and growth processes in non-target crustaceans and beneficial arthropods, e.g. Apis mellifera. Concerns include slow-release methoprene formulations and its environmental breakdown products which mimic an endogenous crustacean hormone and retinoids, respectively. Our primary objective was to evaluate the effect that a chronic methoprene exposure would have on male and female Uca pugnax limb regeneration and molting. After single limb autonomy, limb growth and molt stage were monitored every two days while eyestalk ablation was used to induce proecdysis. Dorsal carapace was collected 6 days post-molt to determine protein and chitin content. In post-molt crabs, methoprene-exposed individuals displayed lower percent gain in body weight. Male crabs lost more weight per body volume than females, took significantly longer to proceed through proecdysis than females exposed to 0.1 microg/L methoprene and exhibited significantly elevated frequency for abnormal limb formation at 1.0 microg/L while females displayed no such trend. Methoprene did not significantly alter extractable exoskeleton protein or chitin content. However, variable water-soluble protein expression increased with exposure at 1.0 microg/L (1 ppb) which contributed to overall variability in total protein content. Our findings suggest that adult male U. pugnax possess greater sensitivity to chronic methoprene exposure during limb regeneration and molting, potentially affecting their post-molt fitness. Furthermore, methoprene has the potential to impact post-molt biomass and exocuticle quality.     

Influence of methoprene and temperature on diapause termination in adult females of the over-wintering solitary bee, Osmia rufa L

Females of Osmia rufa, as most species in this genus, enter an obligatory diapause, overwintering as an imago inside a cocoon until the ensuing spring when after emergence – mating, egg development and oviposition occur. Diapause in this species is initiated in November, undergoes 2 months of a pre-wintering period that is terminated at the end of January, after 1 month of maintenance. In this study, factors that affect the termination of adult diapause in the female of this species were investigated. The experimental material consisted of bees that were brought from nests kept in natural conditions 1 month prior to natural termination of diapause. Three different experimental treatments were planned to evaluate the potential effect of methoprene and temperature on diapause termination. During the 5 day experimental period the first group of females was kept at 4 °C, the second group at 15 °C and the last group of females was kept at 20 °C. All groups of females were treated with methoprene topically at a dose of 200 μg. After methoprene application a significant increase (p < 0.001) in the size of terminal oocytes was recorded in the three experimental groups. However, no changes in the size of terminal oocytes between acetone treated and untreated control groups were observed. The number of oocytes progressively increased following topical application of methoprene compared to non-treated or acetone treated females. In successive applications of 200 μg methoprene gradual changes in ovary and fat body protein concentration were observed. As compared to controls, protein content in ovaries isolated from methoprene-treated females increased, whereas it decreased in fat body. The least differences in oocyte size and protein concentration in ovary and fat body between control groups and with methoprene application occurred at 4 °C. Differences increased and were higher in females kept at 20 °C and increased rapidly after methoprene application. Exposure to increasing temperature regimes accelerated the juvenile hormone (JH) induced termination of diapause. Taken together, our results indicate that temperature may play an important role in termination of diapause in O. rufa, but its role is secondary to that played by JH.     

INCREASE IN LARVAL GUT PROTEOLYTIC ACTIVITIES AND Bti RESISTANCE IN THE DENGUE FEVER MOSQUITO

The bioinsecticide Bacillus thuringiensis var. israelensis (Bti) is increasingly used worldwide for mosquito control. Although no established resistance to Bti has been described in the field so far, a resistant Aedes aegypti strain (LiTOX strain) was selected in the laboratory using field‐collected leaf litter containing Bti toxins. This selected strain exhibits a moderate resistance level to Bti, but a high resistance level to individual Cry toxins. As Bti contains four different toxins, generalist resistance mechanisms affecting mosquito tolerance to different toxins were expected in the resistant strain. In the present work, we show that the resistant strain exhibits an increase of various gut proteolytic activities including trypsins, leucine‐aminopeptidases, and carboxypeptidase A activities. These elevated proteolytic activities resulted in a faster activation of Cry4Aa protoxins while Cry4Ba or Cry11Aa were not affected. These results suggest that changes in proteolytic activities may contribute to Bti resistance in mosquitoes together with other mechanisms.     

Identification of candidate volatiles that affect the behavioural response of the malaria mosquito Anopheles gambiae sensu stricto to an active kairomone blend: laboratory and semi‐field assays

Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) is the most important vector of human malaria in sub‐Saharan Africa, affecting the lives of millions of people. Existing tools such as insecticide‐treated nets and indoor‐residual sprays are not only effective, but also have limitations as a result of the development of resistance to insecticides and behavioural adaptations in biting time. Therefore, novel mosquito‐control tools are needed. Odour‐releasing traps have a potential for both trapping and surveillance purposes. Based on the outcome of ex vivo gene expression assays and in vivo electrophysiological assays on hundreds of volatile organic compounds, 29 ‘candidate behaviourally‐disruptive organic compounds' (cBDOCs) are selected, belonging to 10 chemical categories, to be assayed in the laboratory using dual‐choice olfactometers for the ability to modify the ‘attractiveness’ (i.e. significantly more insects being caught in the associated trap) of a basic volatile blend consisting of ammonia, lactic acid and tetradecanoic acid but without additional carbon dioxide. Compounds that either reduce or increase trap catches by the basic blend in the olfactometer experiments are tested under African conditions in choice experiments in a semi‐field facility in Kenya. The release rates of cBDOCs are determined gravimetrically to allow the calculation of aerial concentrations at the trap outlet. Aerial concentrations in the sub‐p.p.m. range are reported for the first time to influence mosquito behaviour. The results of the olfactometer and semi‐field assays generally correspond. Under semi‐field conditions, three compounds, 3‐heptanol (0.025 p.p.m.), 2‐methylpropanal (0.05 p.p.m.) and 4,5‐dimethylthiazole (0.73 p.p.m.), significantly increase trap catches relative to the basic blend consisting of ammonia, lactic acid, tetradecanoic acid and carbon dioxide. 2‐Acetylthiophene, 2‐nonanone and 2‐phenylethanol decrease the number of mosquitoes caught at all concentrations tested under semi‐field conditions. These compounds hold promise as attractants and spatial repellents to be applied in mosquito control programmes.     

Human skin microbiota and their volatiles as odour baits for the malaria mosquito Anopheles gambiae s.s

Host seeking by the malaria mosquito Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) is mainly guided by volatile chemicals present in human odours. The skin microbiota plays an important role in the production of these volatiles, and skin bacteria grown on agar plates attract An. gambiae s.s. in the laboratory. In this study, the attractiveness of volatiles produced by human skin bacteria to An. gambiae s.s. was tested in laboratory, semi‐field, and field experiments to assess these effects in increasing environmental complexity. A synthetic blend of 10 compounds identified in the headspace of skin bacteria was also tested for its attractiveness. Carbon dioxide significantly increased mosquito catches of traps baited with microbial volatiles in the semi‐field experiments and was therefore added to the field traps. Traps baited with skin bacteria caught significantly more An. gambiae s.s. than control traps, both in the laboratory and semi‐field experiments. Traps baited with the synthetic blend caught more mosquitoes than control traps in the laboratory experiments, but not in the semi‐field experiments. Although bacterial volatiles increased mosquito catches in the field study, trapping several mosquito vector species, these effects were not significant for An. gambiae s.l. It is concluded that volatiles from skin bacteria affect mosquito behaviour under laboratory and semi‐field conditions and, after fine tuning, have the potential to be developed as odour baits for mosquitoes.     

Juvenile hormone (JH) esterase of the mosquito Culex quinquefasciatus is not a target of the JH analog insecticide methoprene

Juvenile hormones (JHs) are essential sesquiterpenes that control insect development and reproduction. JH analog (JHA) insecticides such as methoprene are compounds that mimic the structure and/or biological activity of JH. In this study we obtained a full-length cDNA, cqjhe, from the southern house mosquito Culex quinquefasciatus that encodes CqJHE, an esterase that selectively metabolizes JH. Unlike other recombinant esterases that have been identified from dipteran insects, CqJHE hydrolyzed JH with specificity constant (k(cat)/K(M) ratio) and V(max) values that are common among JH esterases (JHEs). CqJHE showed picomolar sensitivity to OTFP, a JHE-selective inhibitor, but more than 1000-fold lower sensitivity to DFP, a general esterase inhibitor. To our surprise, CqJHE did not metabolize the isopropyl ester of methoprene even when 25 pmol of methoprene was incubated with an amount of CqJHE that was sufficient to hydrolyze 7,200 pmol of JH to JH acid under the same assay conditions. In competition assays in which both JH and methoprene were available to CqJHE, methoprene did not show any inhibitory effects on the JH hydrolysis rate even when methoprene was present in the assay at a 10-fold higher concentration relative to JH. Our findings indicated that JHE is not a molecular target of methoprene. Our findings also do not support the hypothesis that methoprene functions in part by inhibiting the action of JHE.     

Effects of five insect growth regulators on laboratory populations of the North American house-dust mite,Dermatophagoides farinae

The potential of insect growth regulators (methoprene, hydroprene, fenoxycarb, diflubenzuron and triflumuron) to control populations of the North American house-dust miteDermatophagoides farinae (Hughes) was assessed in laboratory bioassays. Methoprene was most effective at suppressing population growth, especially at concentrations of 1.0% (10 000 ppm) and 5.0% (5000 ppm) active ingredient. Hydroprene, structurally related to methoprene, also suppressed house-dust mite populations but not as consistently as methoprene. Fenoxycarb may be effective at controlling house-dust mites but at greater concentrations than were tested. Diflubenzuron and triflumuron, two chitin-synthesis inhibitors, failed to suppress mite numbers and may, in fact, stimulate reproduction in some cases. Almost all concentration of the insect growth regulators were shown to be ineffective when assayed 90 days after treatment.     

Bioaccumulation and Metabolic Effects of the Endocrine Disruptor Methoprene in the Lobster, Homarus americanus

Methoprene is a pesticide that acts as a juvenile hormone agonist.Although developed initially against insects, it has since beenshown to have toxic effects on larval and adult crustaceans.Methoprene was one of several pesticides applied to the WesternLong Island Sound (WLIS) watershed area during the summer of1999; the other pesticides were malathion, resmethrin, and sumethrin.These pesticides were applied as part of a county-by-countyeffort to control the mosquito vector of West Nile Virus. Subsequently,the seasonal lobster catches from the WLIS have decreased dramatically.The lethality of the pesticides to lobsters had been unknown.We studied the effects of methoprene while other investigatorsstudied effects of the other pesticides. We questioned whethermethoprene, through its effects on larvae, adults or both, couldhave contributed to this decline. We found that low levels ofmethoprene had adverse effects on lobster larvae. It was toxicto stage II larvae at 1 ppb. Stage IV larvae were more resistant,but did exhibit significant increases in molt frequency beginningat exposures of 5 ppb. Juvenile lobsters exhibited variationsin tissue susceptibility to methoprene: hepatopancreas appearedto be the most vulnerable, reflected by environmental concentrationsof methoprene inhibiting almost all protein synthesis in thisorgan. Our results indicated that methoprene concentrates in the hepatopancreas,nervous tissue and epidermal cells of the adult lobster. Methoprenealtered the synthesis and incorporation of chitoproteins (cuticleproteins) into adult postmolt lobster explant shells. SDS PAGEanalyses of adult post–molt shell extracts revealed changesin the synthesis of chitoproteins in the methoprene-treatedspecimens, suggesting that methoprene affects the normal pathwayof lobster cuticle synthesis and the quality of the post-moltshell. Although it is likely that a combination of factors ledto the reduced lobster population in WLIS, methoprene may havecontributed both by direct toxic effects and by disrupting homeostaticevents under endocrine control.     

Angling-induced cardiac disturbance of free-swimming largemouth bass (Micropterus salmoides) monitored with heart rate telemetry

The sub‐lethal effects of catch‐and‐release angling have been poorly studied because of the difficulties in monitoring physiological parameters in free‐swimming fish. Laboratory studies provide the opportunity to examine sub‐lethal effects in controlled environments, but do not incorporate site‐specific characteristics. In this study we angled free‐swimming largemouth bass (Micropterus salmoides) equipped with heart rate transmitters to exhaustion using rod and reel, and exposed fish to air for 30 s. Experiments were repeated at four water temperatures (13, 17, 21, and 25°C). These field data were compared with published findings from largemouth bass collected at the same water temperatures in a controlled laboratory setting using Doppler flow probes. Field collected heart rate data increased with increasing water temperatures (Q₁₀ values 1.30–1.37). Pre‐disturbance heart rates were ～30% higher for free‐swimming fish in the field than previously collected laboratory data at the same water temperatures. Fish angled in the field exhausted ～40% more rapidly than fish chased in the laboratory. Maximal heart rate was ～15% higher for free‐swimming fish in the field than for data collected from laboratory restrained fish, but scope for heart rate was reduced by up to 20% in the field, especially at higher water temperatures. Heart rate in free‐swimming fish was highly variable at all times, obscuring clear recovery patterns. Conversely, laboratory cardiac parameters exhibited less variable patterns, peaking clearly following disturbances and recovering in about 135 min, independent of water temperature. Based upon these findings, we suggest that comprehensive studies incorporating both laboratory and field experiments are needed for truly understanding the effect of catch‐and‐release angling on fish.