Susceptibility of Culex quinquefasciatus (Diptera: Culicidae) in Southern Louisiana to Larval Insecticides.

Mosquito control districts conduct rigorous insecticide treatments against both larval and adult Culex quinquefasciatus Say (Diptera: Culicidae), the primary vector of West Nile virus in the southern United States. However, the development of resistant populations of Cx. quinquefasciatus in response to extensive larvicide or adulticide applications has been demonstrated repeatedly across the world. Examining changes in insecticide susceptibility in treated field areas can help inform mosquito control districts as to whether or not their treatments remain effective. We hypothesized that frequent insecticide applications for the control of mosquitoes in East Baton Rouge Parish, Louisiana, lowered susceptibility of wild Cx. quinquefasciatus to larvicides. Larvicide susceptibility was measured using Lysinibacillus sphaericus, spinosad, and temephos in populations of Cx. quinquefasciatus sampled from sites in three Parishes where frequencies of insecticide applications varied, and frequencies of resistance were measured relative to a susceptible reference colony. Susceptibility to these larvicides was widespread, although fourfold resistance to the organophosphate temephos was detected at one site in East Baton Rouge Parish in the spring of 2016, which increased to eightfold resistance by the end of the mosquito season. Activities of esterases were found to be elevated in wild, temephos-resistant mosquitoes, indicating the potential role of these enzymes as a mechanism of resistance. The results of this study provide a baseline of comparison for future measurements of susceptibility in Cx. quinquefasciatus in Louisiana, and may help inform local mosquito control districts as to the effectiveness and sustainability of their insecticide programs.

Limited impacts of truck-based ultra-low-volume applications of mosquito adulticides on mortality in honey bees (Apis mellifera).

Adulticides applied against mosquitoes can reduce vector populations during times of high arbovirus transmission. However, impacts of these insecticides on pollinators and other non-target organisms are of concern to mosquito control professionals, beekeepers and others. We evaluated mortality of Culex quinquefasciatus and Apis mellifera when caged insects were exposed to low and high label rates of four common adulticides (Aqua-Pursuit™ [permethrin], Duet® [prallethrin + sumithrin], Fyfanon® [malathion] and Scourge® [resmethrin]) at six distances up to 91.4 m from a truck-mounted ultra-low-volume sprayer. Honey bee mortality was both absolutely low (61 m had limited impacts on honey bee mortality while providing effective mosquito control.

Effects of chlorantraniliprole and thiamethoxam rice seed treatments on egg numbers and first instar survival of Lissorhoptrus oryzophilus (Coleoptera: Curculionidae).

Effects of treatment of rice seeds with an anthranilic diamide, chlorantraniliprole, and a neonicotinoid, thiamethoxam, on egg laying and first instar survival in rice water weevil, Lissorhoptrus oryzophilus Kuschel, were examined under greenhouse conditions. Exposure of adult weevils to rice (6-7 leaf stage) grown from seeds treated with chlorantraniliprole and thiamethoxam resulted in reduction in numbers of eggs and first instars. The low egg numbers by adults exposed to chlorantraniliprole-treated plants was confirmed as a sublethal effect on adults: adult survival was not impacted after 4 d of feeding on foliage from chlorantraniliprole-treated plants but the number of eggs laid by these weevils was reduced when released on untreated plants. Furthermore, a comparison of first instar emergence from chlorantraniliprole-treated plants and from untreated plants infested with weevils previously exposed to this chemical suggested that chlorantraniliprole was also reducing egg or first instar survival. In contrast, adults that fed on foliage from thiamethoxam-treated plants showed increased mortality. Possible sublethal effects of thiamethoxam on the number of eggs laid by adults were investigated by infesting untreated plants with weevils that survived exposure to thiamethoxam via foliar feeding (7 microg active ingredient/seed). Prior exposure to thiamethoxam through adult feeding reduced egg numbers. However, potential larvicidal or ovicidal effects of thiamethoxam seed treatments could not be detected in this study because of low first instar emergence from both thiamethoxam-treated plants and from untreated plants infested with weevils previously exposed to this chemical. These experiments revealed that the two seed treatments accomplish weevil control in different ways.

Susceptibility to insecticides used for control of Piezodorus guildinii (Heteroptera: Pentatomidae) in the United States and Brazil.

Limited information exists on the insecticide susceptibility of redbanded stink bug, Piezodorus guildinii (Westwood) (Heteroptera: Pentatomidae), despite its impact on soybean, Glycine max (L.) Merr., production in Brazil and the United States. Therefore, this study set out to 1) determine baseline levels of susceptibility to currently recommended pesticides using topical and vial bioassays, 2) determine the levels of esterase activity in populations in the United States and Brazil, and 3) compare control among products in field trials. In topical bioassays conducted in the United States using technical grade materials, the LC50 values of lambda-cyhalothrin, acephate, and methamidophos were 4-25, 141-295, and 40-151 ng per insect, respectively. The LC50 values of imidacloprid and thiamethoxam were 11 and 27 ng per insect, respectively. In vial bioassays conducted in the United States using technical grade materials, the LC50 values of cypermethrin, acephate, and methamidophos were 0.4-0.9, 3.8, and 1.6 microg per vial, respectively. In topical bioassays conducted in Brazil by using commercially formulated products, the LC50 values of acephate, methamidophos, endosulfan, and imidacloprid were 0.90-1.9, 0.4-0.6, 1.5-6.6, and 0.2-0.3 microg per insect, respectively. In vial bioassays conducted in Brazil using commercially formulated products, the LC50 values of endosulfan, methamidophos, and lambda-cyhalothrin were 4-32 and 2-24 microg/cm2 for thiamethoxam and imidacloprid. Esterase activity in Louisiana (United States) populations ranged from 251 to 658 nmol alpha-naphthol formed/min/mg protein. Esterase activity levels in Londrina (Brazil) populations averaged 163 nmol/min/mg. In field tests, P. guildinii in Louisiana were controlled by organophosphates thiamethoxam and imidacloprid and in Brazil, with combinations of neonicotinoids and pyrethroids.

Selection and life history traits of tebufenozide-resistant sugarcane borer (Lepidoptera: Crambidae).

Varying susceptibility to tebufenozide was recorded in the sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), collected from Louisiana sugarcane locations with different selection pressures. Results from diet incorporation bioassays with tebufenozide indicated significant increases in LC50 (3.78-fold) and LC90 (7-fold) values for a colony from Duson (DU), an area with higher selection pressure, compared with a colony from Alexandria (ALEX), an area with no selection pressure. Differences were not detected in LC50 values among colonies from areas where use of tebufenozide was discontinued or rotated with other chemistries. Selections with tebufenozide of DU larvae over 12 generations resulted in a highly resistant colony (DU-R) with 27.1- and 83.3-fold increases in LC50 and LC90 values, respectively. Comparison of pupal weight, days to pupation, and emergence after exposure to an equitoxic (LC20) concentration of tebufenozide revealed a decrease in pupal weight (34 and 33% for males and females, respectively), and an increase in days to pupation (47 and 40% for males and females, respectively), and emergence (43 and 33% for males and females, respectively) for the DU-R colony compared with the parent DU colony. Fecundity of DU-R females decreased to 72 eggs per female compared with 180 (DU) and 261 (ALEX). Egg viability of the ALEX and DU colonies was 61 and 56%, respectively, whereas only 27% of eggs laid by females from the DU-R colony hatched. These results are discussed in terms of their practical implications for control of D. saccharalis in Louisiana sugarcane.

Reduced susceptibility to tebufenozide in populations of the sugarcane borer (Lepidoptera: Crambidae) in Louisiana.

Susceptibility of the sugarcane borer, Diatraea saccharalis (F.), to tebufenozide was measured using a feeding bioassay, and values obtained were compared with baselines generated before the use of this insecticide in Louisiana sugarcane, Saccharum spp. Results from our study suggest that susceptibility to tebufenozide is decreasing in field-collected sugarcane borers. Inflections in the log dosage-probit lines were detected for many of the field collections, indicating increased heterogeneity within these populations in response to tebufenozide. Where appropriate, probit transformation was used to estimate susceptibility, and significant differences (1.6 - 2.7-fold) were measured in LC50 values between some field-collected cohorts and the previously measured baseline. In addition, a discriminating concentration (0.5 ppm) was used to estimate resistance frequencies in cohorts for which probit transformation was not appropriate. Results from these tests suggest that frequencies of resistance were high (49% in one cohort) in populations from some locations. Lighter weight pupae of the survivors from one of the more resistant cohorts suggests that tebufenozide resistance mechanisms may have a biological cost in terms of ecological fitness at early stages of resistance development. As a result of continued resistance monitoring, alternation of management chemistry is expected to help preserve this valuable sugarcane integrated pest management tactic.

Horn fly (Diptera: Muscidae) resistance to organophosphate insecticides.

Insecticidal ear tags impregnated with organophosphate (OP) insecticides were used each year from 1989 to 1998 at Rosepine, LA. Weekly fly counts were conducted to evaluate control efficacy of the treatments, and bioassays were conducted at least twice per year to measure fly susceptibility to OP and pyrethroid insecticides. Between 1989 and 1992, the efficacy of 20% diazinon-impregnated ear tags was reduced from >20 to just 1 week of control. A high risk of control failure was observed when a resistance frequency of approximately 5% was measured in pre-season bioassays. Resistance to diazinon, fenthion, ethion, pirimiphos-methyl, and tetrachlorvinphos was observed. Esterase activity toward alpha-naphthyl acetate was significantly higher in flies collected at Rosepine in 1997 than in flies from a laboratory colony and from a susceptible field population.

Susceptibility of Diatraea saccharalis (Lepidoptera: Crambidae) to tebufenozide.

Dosage-mortality baselines were determined for first and newly molted third instars of the sugarcane borer, Diatraea saccharalis (F.), with tebufenozide incorporated into an artificial diet. Using death as an endpoint for these assays, LC50 and LC90 values were estimated to be 0.17 and 0.41 ppm for the first and 0.87 and 3.67 ppm for third instars, respectively. Developmental effects also were observed and were defined to include mortality as well as physical impairment and retarded or prolonged development with absence of feeding. Concentrations causing developmental effects in treated larvae (EC50 and EC90) were 5.8- and 3.8-fold lower than the LC50 and LC90, respectively. The critical time of exposure to tebufenozide was determined for developmentally synchronous third instars to be less than 12 h postecdysis. Mortality through adult emergence significantly decreased from 92% (treatment at 0 h postecdysis) to 30% (treatment at 12 h postecdysis), when exposed to tebufenozide at the EC50. For surviving larvae, length of development time until pupation was significantly increased and female pupal weight significantly decreased when larvae were treated at 0 h postecdysis into the third instar. In addition, to modifying the traditional ways of assessing mortality 0-3 d postapplication, this study points out the value of using other approaches to pesticide assessment, especially where insect growth regulators are involved.

Selection, egg viability, and fecundity of the sugarcane borer (Lepidoptera: Crambidae) with tebufenozide.

Two separate attempts to select the sugarcane borer, Diatraea saccharalis (F.), for resistance to tebufenozide were unsuccessful. Both selected colonies were lost after the fourth generation due to a lack of oviposition. Differences were not detected in fecundity or percent egg viability for 5-d-old third instars exposed to concentrations (EC5, EC15, and EC30) of tebufenozide for 7 d. Decreases (P < or = 0.01) in mean female pupal weights were detected in larvae exposed to EC15 and EC30 concentrations. An ovicidal impact using serial dilutions of tebufenozide (10, 100, and 200 ppm) also was detected. Percent viability was reduced from 98% for untreated eggs to 61% for eggs dipped in 10 ppm and below 6% for eggs dipped in > or = 100 ppm. Eggs treated with 200 ppm did not hatch. Though some embryonic development was observed on eggs treated with the high concentrations (100 and 200 ppm), sclerotization of head capsule was not apparent. The ovicidal property of tebufenozide may enhance its effectiveness in controlling populations of the D. saccharalis on an area-wide basis. Fecundity and egg viability were affected in later generations of selection; however, separate studies assessing individuals that were exposed to sublethal concentration (EC5, EC15, and EC30) of tebufenozide as third instars for 7 d in one generation did not detect differences.

Characterization of esterases associated with profenofos resistance in the tobacco budworm, Heliothis virescens (F.).

The utility of microplate and electrophoretic assays for detection of biochemical and physiological mechanisms underlying resistance to profenofos in the tobacco budworm, Heliothis virescens (F.), was assessed. Esterase (EST) activities were significantly higher in profenofos-resistant than -susceptible larvae, and activities were highly correlated (r(2) = 0.87) with resistance to profenofos. Both qualitative and quantitative variation was observed in electrophoretic gels stained with alpha- and beta-naphthyl acetates. Staining of ESTs was more intense with resistant larvae than those from a susceptible strain. In addition, a band (designated A') was expressed in larvae from profenofos-resistant strains, but not in larvae from an insecticide-susceptible strain. The frequency of expression of A' increased following selection with profenofos and was detected in 100% of the individuals from a profenofos-selected strain. The appearance of this band coincided with the decreased expression of a second band (designated A). A similar pattern (overexpression of A' and underexpression of A) also was observed in larvae from field-collected strains. Finally, reduction in the activity or the sensitivity of acetylcholinesterase to inhibition by chlorpyrifos oxon was observed in laboratory-selected and field-collected larvae that expressed resistance to profenofos. These results suggest that microplate and electrophoretic assays can be utilized as complementary tools for detecting and monitoring profenofos resistance in H. virescens.

Activation of sodium channels and inhibition of [3H]batrachotoxinin A-20-alpha-benzoate binding by an N-alkylamide neurotoxin.

BTG 502 [(2E,4E)-N-(1,2-dimethyl)-propyl-6-(5-bromonaphth-2-yl)-hexa -2,4- dienamide], a synthetic analog of insecticidal amides isolated from Piper species, stimulated 22Na+ uptake into mouse brain synaptoneurosomes in the presence of saturating concentrations of Leiurus quinquestriatus venom but had no effect on sodium uptake in the absence of venom. In the presence of Leiurus venom, half-maximal stimulation was achieved at a BTG 502 concentration of 1.7 microM, whereas maximal stimulation (2.3-fold greater than nonspecific uptake) was observed at 50 microM. In the absence of other modifiers, BTG 502 inhibited batrachotoxin (BTX)-dependent sodium uptake, producing 50% inhibition at 2 microM. In the presence of Leiurus venom, BTG 502 was a partial inhibitor of BTX-dependent 22Na+ uptake, producing half-maximal inhibition at 1.5 microM. The levels of residual BTX-dependent sodium uptake and maximal BTG 502-dependent sodium uptake measured in the presence of Leiurus venom were identical. BTG 502 inhibited the specific binding of [3H]batrachotoxinin A-20-alpha-benzoate (BTX-B) to the activator recognition site (site 2) of sodium channels in these preparations, producing half-maximal inhibition at 2 microM and maximal inhibition at 30 microM. Equilibrium analysis showed that BTG 502 was an apparent competitive inhibitor of [3H]BTX-B binding, producing a concentration-dependent decrease in the affinity of sodium channels for this ligand without affecting binding capacity. Kinetic analysis demonstrated that BTG 502 slowed the rate of formation of the ligand-receptor complex but did not alter the rate of dissociation of this complex. The effects of BTG 502 on 22Na+ uptake and [3H]BTX-B binding are consistent with the action of this compound as an antagonist at the activator recognition site of the voltage-sensitive sodium channel in the absence of Leiurus venom and as a partial agonist at this site in the presence of Leiurus venom. These results suggest that the N-alkylamides represent a novel chemical class of neurotoxins that act at site 2 of the sodium channel.

Patterns of epoxide metabolism by epoxide hydrolase and glutathione S-transferase associated with age and genotype in Drosophila melanogaster.

Epoxide hydrolase and glutathione S-transferase activities toward trans- and cis-stilbene oxides were measured in 3 strains of Drosophila melanogaster. Differences in age dependence, substrate selectivity and subcellular location were detected suggesting the presence of multiple forms of these enzymes. In addition, interstrain differences indicate the presence of genetic variation for epoxide hydrolase and glutathione transferase activities. These results illustrate a potential use of these assays in D. melanogaster to complement existing tests (e.g. recessive lethal tests or Ames assays) for evaluating the relationship between epoxide hydrolase and glutathione S-transferase levels and the genotoxicity of epoxides.