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Determination of acute mortality in adults and sublethal embryo responses of Palaemonetes pugio to endosulfan and methoprene exposure.

Adult grass shrimp (Palaemonetes pugio) were exposed to endosulfan or methoprene for 96 h and LC(50) values were calculated. Male and female P. pugio cohorts were also exposed to endosulfan for 96 h in an attempt to determine potential differences in sensitivity between the sexes. Results from the methoprene exposure indicated that this pesticide was not acutely toxic to adult grass shrimp at 1 mg l(-1). Due to the lack of sensitivity, sex specific tests with methoprene were not performed. The calculated LC(50) for a population of grass shrimp, including both males and females exposed to endosulfan, was 0.62 microg l(-1). The LC(50) determinations for the sex specific tests were 0.92 microg l(-1) for males and 1.99 microg l(-1) for females. Following these acute exposures, reproductively active grass shrimp were chronically exposed to 200 ng l(-1) endosulfan or 1 mg l(-1) methoprene and were allowed to produce embryos. The resulting embryos were assessed for potential sublethal toxicity. There were no observed differences in the percent successfully hatching or larval mortality 3-days post hatch among the treatments. However, endosulfan treated embryos had a significantly increased hatching time (9.76 days compared to 8.72 days in controls). Methoprene treated embryos also took longer to hatch (9.55 days), but this delay was not significantly different from controls. These findings suggest that endosulfan may preferentially affect male grass shrimp and exposed female grass shrimp may produce embryos with delayed hatching times.

Animals↗

Effectiveness and residual activity comparison of granular formulations of insect growth regulators pyriproxyfen and s-methoprene against Florida mosquitoes in laboratory and outdoor conditions.

Effectiveness and residual activity tests of granular formulations of 2 insect growth regulators (IGRs), s-methoprene and pyriproxyfen, against laboratory-reared larvae of 5 colonized mosquitoes, Aedes aegypti, Aedes albopictus, Aedes taeniorhynchus, Anopheles quadrimaculatus, and Culex nigripalpus, were conducted in the laboratory and outdoors in plastic tubs. Culex quinquefasciatus was exposed to these two IGRs in the laboratory only. Each IGR formulation was applied at 0.02 and 0.05 ppm active ingredient (Al) against 5 of the 6 mosquito species both in the laboratory and the outdoor evaluations, whereas Cx. quinquefasciatus was exposed to 0.2 and 0.4 ppm AI of s-methoprene, and 0.1 and 0.2 ppm AI of pyriproxyfen in the laboratory. s-Methoprene at 0.02 and 0.05 ppm AI resulted in variable levels (<39-100%) of inhibition of adult emergence in the 5 species monitored for 6 weeks after treatment under both test conditions. Aedes taeniorhynchus was the most susceptible to s-methoprene in terms of initial and residual activity. Culex quinquefasciatus and Ae. albopictus were the most tolerant to s-methopene, with maximum emergence inhibitions amounting to 84% in Cx. quinquefasciatus at 0.4 ppm and 44.3% in Ae. albopictus at 0.05 ppm during the 1st week in the laboratory. Pyriproxyfen at comparable treatment rates to s-methoprene caused very high levels (>80-100% in most cases) of initial and residual emergence inhibitions of the tested species in the laboratory as well as outdoors. In several species, pyriproxyfen induced complete inhibition of adult emergence for several weeks after treatment, even at the lower rate of 0.02 ppm. The World Health Organization has recently recommended the use of pyriproxyfen for the control of some mosquito species at specified rates in certain habitats.

Aedes↗

Comparative efficacy of the combination fipronil-(S)-methoprene and the combination permethrin-imidacloprid against Dermacentor reticulatus, the European dog tick, applied topically to dogs.

This study compared the efficacy of two of the most widely veterinary-dispensed topical products for control of ticks on dogs: fipronil-(S)-methoprene and imidacloprid-permethrin. Eighteen healthy beagle dogs of both sexes were divided into three groups of six dogs. Group 1 served as the untreated control. Dogs in group 2 were treated with fipronil 10% + (S)-methoprene 9% w/v applied once on day 0, and those in group 3 were treated with imidacloprid 8.8% and permethrin 44% w/v, applied once on day 0. All dogs were infested with approximately 50 unfed Dermacentor reticulatus ticks on days 1, 7, 14, 21, 28, 35, and 42, and ticks remaining were counted and removed 48 hours after each infestation. The 48-hour efficacy of the fipronil-(S)-methoprene combination remained at 100% at all assessment points through and including day 37, declining to 95.30% on day 44. The 48-hour efficacy of the imidacloprid-permethrin combination peaked at 86.46% on day 9, decreased to 73.37% by the third week, and fell to 63.53% by the end of the study (day 44). Significantly (P < .05) fewer ticks were recovered from dogs treated with fipronil-(S)-methoprene or imidacloprid-permethrin than from untreated control dogs at each infestation. Notably, the dogs treated with fipronil-(S)-methoprene had significantly (P < .05) lower tick counts at every assessment than the dogs treated with imidacloprid-permethrin.

Administration, Topical↗

Effect of the insect growth regulator methoprene on the ovipositional behavior of Aedes aegypti and Culex quinquefasciatus.

Laboratory and field tests were conducted to determine the attractiveness of the insect growth regulator methoprene to ovipositing mosquitoes when presented at concentrations used in mosquito control programs. Laboratory experiments indicated that ovipositing Culex quinquefasciatus and Aedes aegypti were not attracted to methoprene. In field experiments, Culex stigmatosoma and Cx. quinquefasciatus larval dip counts in experimental ponds treated with methoprene briquets were not significantly different from untreated control ponds. Water taken from these methoprene-treated ponds was not attractive when compared with water taken from untreated ponds to ovipositing Cx. quinquefasciatus in laboratory experiments. These studies provide strong evidence that methoprene is not attractive to ovipositing mosquitoes at concentrations within the range of field applications.

Aedes↗

Evaluation of Methoprene (Altosid) and Diflubenzuron (Dimilin) for control of mosquito breeding in Tezpur (Assam).

Insect growth regulators (IGRs) namely, Isopropyl (E-E)-(RS)-11-methoxy-3,7,11-trimethyldodeca-2, 4-dinoate (Methoprene) and 1-(4-cyclophenyl)-3-(2,6-diflerobenzoyl) urea (Diflubenzuron) were evaluated against mosquito larvae in laboratory as well as in different breeding habitats in Tezpur, Assam. LC90 values of diflubenzuron against Culex quinquefasciatus and Aedes albopictus were 0.0022 and 0.0027 ppm respectively, while it was 0.0027 and 0.0022 ppm respectively in case of methoprene. However, LC50 values of both the IGRs were almost same in case of Ae. albopictus and Cx. quinquefasciatus (varies between 0.0009 and 0.0011 ppm). In case of methoprene, maximum mortality was observed in pupal stage though the exposure was given in all the cases to the III instar larvae. Field trials were conducted in cemented drains, small ponds and ditches. At 0.2 ppm (0.020 kg/ha) both diflubenzuron and methoprene were found to eliminate 92-96 per cent Culex and Anopheles larvae. Methoprene and diflubenzuron were found equally effective for control of mosquito breeding in different breeding habitats and provide better efficacy than conventional larvicides and biocides.

Aedes↗

Inhibition of the larval ecdysis and emergence behavior of the parasitoid Cotesia congregata by methoprene.

Parasitism of the tobacco hornworm, Manduca sexta, by the braconid wasp Cotesia congregata, induces developmental arrest of the host in the larval stage. During the final instar of the host, its juvenile hormone (JH) titer is elevated, preventing host metamorphosis. This study investigated the effects of hormonal manipulation of the host on the parasitoid's emergence behavior. The second larval ecdysis of the wasps coincides with their emergence from the host, and application of the juvenile hormone analogue methoprene to day 4 fifth instar hosts either delayed or totally suppressed the subsequent emergence of the wasps. Effects of methoprene were dose-dependent and no parasitoids emerged following treatment of host larvae with doses >50 &mgr;g. Parasitoids which failed to emerge eventually succumbed as unecydsed pharate third instar larvae in the hemocoel of the host. Effects of host methoprene treatment on parasitoid metamorphosis were also assessed, and metamorphic disruption occurred at much lower dosages compared with doses necessary to suppress parasitoid emergence behavior. The inhibitory effect of methoprene on parasitoid emergence behavior appears to be mediated by effects of this hormone on the synthesis or release of ecdysis-triggering hormone (ETH) in the parasitoid, the proximate endocrine cue which triggers ecdysis behavior in free-living insects. ETH accumulated in the epitracheal Inka cells of parasitoids developing in methoprene-treated hosts, suggestive of a lack of hormone release. Thus, the hormonal modulation of parasitoid emergence behavior appears to be complex, involving a suite of hormones including JH, ecdysteroid, and peptide hormones.

Journal Article↗

A solid phase extraction reversed-phase HPLC method for the simultaneous determination of methoprene, permethrin and selected metabolites in rat plasma and urine.

A method was validated and applied for the analysis of the insect growth regulator methoprene [Isopropyl (2E,4E)-11-methoxy-3,7,11-trimethyldodeca-2,4-dienoate], its metabolite methoprene acid, the insecticide permethrin [3-(2,2-dichloro-ethenyl)-2,2-dimethylcyclopropanecarboxylic acid(3-phenoxyphenyl)methylester], and two of its metabolites, m-phenoxybenzyl alcohol and m-phenoxybenzoic acid, in rat plasma and urine using solid-phase extraction and reversed-phase high performance liquid chromatography. The analytes were separated using gradient of 55-100% acetonitrile in water (pH 4.0) at a flow rate ranging between 0.6 and 1.0 mL/min over a period of 20 min, and UV detection at 210 and 254 nm. The retention times ranged from 7.3 to 18.4 min. The limits of detection ranged between 50 and 100 ng/ml, while limits of quantitation were 100-150 ng/mL. Average percentage recovery of five spiked plasma samples was 83.6 +/- 3.9, 80.1 +/- 5.4, 82.1 +/- 4.4, 83.7 +/- 3.9 and 83.1 +/- 4.7, and from urine 79.3 +/- 4.3, 82.0 +/- 5.4, 80.7 +/- 4.2, 78.9 +/- 5.7 and 83.9 +/- 4.5 for methoprene, methoprene acid, permethrin, m-phenoxybenzyl alcohol and m-phenoxybenzoic acid, respectively. The method was linear and reproducible over the range of 100-1000 ng/mL. This method was applied to analyze the above chemicals and metabolites following their combined administration in rats.

Animals↗

The juvenile hormone analogue, methoprene, inhibits ecdysterone induction of small heat shock protein gene expression.

The small heat shock protein (hsp) genes of Drosophila are expressed in cultured cells in response to the moulting hormone, ecdysterone. We show here that juvenile hormone (JHIII) and the juvenile hormone analogue, methoprene, inhibit that induction in a dose-dependent manner. Heat shock induction is not inhibited. In transient expression studies using S3 line cells transfected with EcRE-CAT constructs, methoprene inhibition was found to require a 2-hr pretreatment (before ecdysterone addition), and methoprene's continued presence was essential. Farnesol, farnesyl acetate, and retinoic acid did not cause inhibition. Several models of methoprene inhibition are discussed.

Animals↗

Expression of nuclear receptor-transcription factor genes during Aedes aegypti midgut metamorphosis and the effect of methoprene on expression.

Exposure of mosquito 4th instars to the juvenile hormone analogue methoprene prevents the emergence of adults by interfering with metamorphosis. One metamorphic processes that is disrupted is midgut remodeling. To investigate the molecular mechanisms by which this occurs, the pattern of transcription factor gene expression during the Aedes aegypti (L.) 4th instar was investigated by the method of real time PCR. The results indicate that in untreated larvae, expression of transcription factors genes AHR3 and AaE75B increases within 24h after the last larval-larval molt, transcription of AaEcR-B, AaUSP-a and AassFTZ-F1 increases approximately 24h later, and transcription of AaE75A increases just before the larval-pupal molt. There is uniform expression of AaUSP-b throughout the 4th instar. The effect of methoprene exposure on transcription factor gene expression during midgut remodeling was investigated. The results indicate that, in a dose and stage dependent manner, methoprene affects increases in expression that normally occur during midgut remodeling. The coincident effects of methoprene on metamorphic midgut remodeling and on transcription factor gene expression suggests that the two processes are related.

Aedes↗

Trypanosoma cruzi: methoprene is a potent agent to sterilize blood infected with trypomastigotes.

The effects of methoprene, a juvenile hormone analogue (JHA), on Trypanosoma cruzi bloodstream trypomastigotes (Tulahuen strain, Tul 2 stock) were studied. It was observed that 150microM of methoprene in in vitro experiments cause cellular death of T. cruzi. In contrast, methoprene was not able to clear bloodstream trypomastigotes in in vivo experiments, but it was observed a decrease of parasitemia levels of infected mice treated with 200microg of methoprene/mouse/day during 5 days. According to these results and the low toxicity of methoprene, we suggest that this compound will serve as an effective agent to sterilize blood for transfusions.

Animals↗

Effectiveness of methoprene, an insect growth regulator, against temephos-resistant Aedes aegypti populations from different Brazilian localities, under laboratory conditions.

The susceptibility of Aedes aegypti (L.) larvae from several Brazilian populations to the juvenile hormone analog methoprene and the organophosphate insecticide temephos were investigated. Populations from Natal (northeastern region), Macapá (northern region), and Jardim América, Rio de Janeiro (southeastern region) are temephos-resistant (RR90 = 24.4, 13.3, and 15.8, respectively), whereas populations from Presidente Prudente (southeastern region) and Porto Velho (northern region) exhibit only an incipient temephos-altered susceptibility status (RR90 = 1.8 and 2.6, respectively). Biochemical assays revealed alterations of the enzymes implicated in metabolic resistance, glutathione S-transferase, mixed function oxidases and esterases, among these populations. Dose-response assays showed at most a low resistance to methoprene of all populations tested, irrespective of their temephos resistance level. However, sequential exposure of Macapá and Natal populations to temephos and methoprene indicated a potential cross-resistance when larvae are exposed to both insecticides. Nevertheless, susceptibility of the Brazilian Ae. aegypti populations to methoprene alone suggests this insect growth regulator could substitute for temephos in the control of the dengue vector in the country.

Acetylcholinesterase↗

Effects of Ecdysterone and the juvenile hormone analogue methoprene on protein, RNA and DNA synthesis in wing discs of Calliphora vicina.

The protein, RNA and DNA content of wing discs increase exponentially during the last larval instar. Biosynthesis of protein, RNA and DNA was studied by injecting labelled precursors into larvae or white prepupae and measuring the incorporation in the wing discs dissected out after an appropriate time. Protein synthesis is stimulated by ecdysterone, methoprene has no effect. Biosynthesis of rRNA is increased in wing discs of white prepupae after ecdysterone or methoprene injection. Methoprene inhibits the synthesis of mRNA, while ecdysterone has no clear-cut effect within the limits of our method (gel electrophoretic analysis). Ecdysterone and methoprene have no detectable influence on incorporation of thymidine into DNA, but the incorporation label from uridine into DNA is diminished; this effect may be due to changes in the precursor pool.

Animals↗

Efficacy of broadcast and perimeter applications of S-methoprene bait on the red imported fire ant in grazed pastures.

The red imported fire ant, Solenopsis invicta (Buren), is a major pest in the United States because of its painful sting. Toxic bait has been an important management tool against fire ants, but site registrations prohibit applications of most baits on grazed pastures. Extinguish, containing the insect growth regulator methoprene, was selected for this study because it has a broad site registration that includes grazed pastures. The primary objective of this research was to evaluate the efficacy for control of red imported fire ants by using broadcast applications of methoprene bait at a label rate of 1,121 g/ha versus applications around the perimeter of a target area at the reduced rate of 280 g/ha. Grazed pastures in Lee County, Alabama, and Chambers County, Alabama, were selected for this study, with broadcast treatments, perimeter treatments, and controls replicated three times at each site. All mounds were counted and rated using the USDA population index before applications and then at 8 and 16 wk posttreatment. Perimeter applications did not significantly reduce S. invicta mound abundance, but bait treatments significantly reduced mound abundance at 16 wk posttreatment at site 1 where applications were conducted in early evening. However, broadcast applications were not effective at site 2 where treatments were conducted in early morning with warmer temperatures. Emergence of winged alates was observed at 12 wk posttreatment, followed by a high density of incipient mounds that may have masked the full treatment effect of methoprene applications at site 2. Methoprene bait was effective in reducing abundance of S. invicta only when full label rates were applied.

Agriculture↗

Effects of methoprene on oviposition by Aedes japonicus and Culex spp.

The mosquito larvicide methoprene is a juvenile growth hormone mimic that is widely used to control mosquitoes. This chemical disrupts normal mosquito development, drastically inhibiting emergence from the pupal to the adult stage. If the presence of methoprene attracts or deters mosquitoes from ovipositing it could have implications for mosquito control. This study evaluates whether methoprene attracts or deters mosquitoes likely to oviposit in catch basins. In a field experiment, methoprene formulated as liquid larvicide did not affect oviposition of either Culex spp. or Aedes japonicus in 19-1 plastic buckets.

Aedes↗

Investigation of the relationship between s-methoprene and deformities in anurans.

Evidence that amphibian deformities are on the rise has prompted a number of researchers to question a variety of natural and man-made substances, including chemicals such as s-methoprene (trademark Altosid), an insect growth regulator applied to water bodies for mosquito control. Despite conclusions reached by the U.S. Environmental Protection Agency and a considerable body of scientific evidence demonstrating that it is unlikely that s-methoprene or any of its degradation products are causing the high rates of deformities found in Minnesota and other states, concerns persist about potential negative impacts of s-methoprene on nontarget species, specifically anurans. Water analyses in field and laboratory conditions and a comparison of reported Altosid use with reported frog deformities in Minnesota demonstrate that a connection between frog deformities and Altosid use is unlikely. These results indicate that factors other than s-methoprene and its degradation products are contributing to the recent outbreak of frog deformities.

Animals↗

Reproductive effects in German cockroaches by ecdysteroid agonist RH-0345, juvenile hormone analogue methoprene and carbamate benfuracarb.

Blatta germanica is the more prevalent cockroach species in Algeria. In the present study, we tested the effect on reproduction in B. germanica of two insect growth regulators, RH-0345, a benzoylhydrazine analogue that mimics the action of 20-hydroxyecdysone, and methoprene, one of the most commercially important juvenile hormone analogues, and a novel carbamate insecticide, benfuracarb. The compounds were applied topically (10 and 20 microg/insect for RH-0345, and 1 and 10 microg/insect for methoprene) or orally administrated (at 2% for benfuracarb) on newly emerged females and evaluated on reproductive events during the adult life (2, 4 and 6 days). Treatment with RH-0345 and benfuracarb reduced significantly the number of oocytes, the size and the volume of the basal oocyte during the experimental period. Methoprene distorted the ovarian development since it caused a significant reduction in the number of oocytes at 2, 4 and 6 days for the two tested doses, and an increase in oocyte size at 2, 4 and 6 days with 1 microg and a decrease with 10 microg. In a second series of experiments, the effects of these compounds were assayed on the ovarian proteins. Data from biochemical analysis revealed that RH-0345 and benfuracarb reduced the ovarian amounts of proteins, while treatment with methoprene increased it during the sexual maturation.

Administration, Topical↗

Evaluation of methoprene, temephos and Bacillus thuringiensis var. israelensis against Coquillettidia perturbans larvae in Minnesota.

Temephos, Bacillus thuringiensis var. israelensis (B.t.i.) and methoprene were tested for larval control of Coquillettidia perturbans. Neither temephos nor B.t.i. treatments at their maximum recommended dosages consistently reduced larval numbers. Larval emergencies were reduced 99% in test plots treated with experimental, controlled release methoprene briquets (Altosid SR-10). Breeding sites of Cq. perturbans in a 5,000 km2 area were treated using methoprene briquets in the 1984 season. Adult populations in a 7,700 km2 area were 2.5 times higher in untreated areas than treated areas. Methoprene can effectively control larval populations of Cq. perturbans.

Animals↗

Toxicity of methoprene to all stages of the salt marsh copepod, Apocyclops spartinus (Cyclopoida).

The toxicity of methoprene to the salt marsh copepod Apocyclops spartinus was evaluated and compared with sensitivity of mosquito larvae. All stages of the life cycle were tested at concentrations ranging from 0.1 to 10.0 ppm. Eggs and the earliest hatched stages, nauplius I-III were most sensitive to methoprene, with little mortality seen in the later stages. Toxic effects were manifested as death, or failure of eggs to hatch, however, no extensions of the life cycle were observed. In general the copepods were resistant at concentrations of methoprene used to control mosquitoes. Early nauplii, however, did show some mortalities to methoprene concentrations near the lower margins of mosquito susceptibility. This might lead to transient decreases in copepod population growth rates, but not necessarily to decreases in their standing populations.

Aedes↗