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European deer keds, Lipoptena cervi (Linnaeus, 1758), are hematophagous ectoparasitic flies known to bite cervids and noncervids, including humans. To prevent deer keds from landing and biting hosts, 5 commercially available insect repellents (DEET, IR3535, oil of lemon eucalyptus (OLE), picaridin, and permethrin) and water control were evaluated to determine repellency efficacy and postexposure mortality of deer keds. While there was a significant difference between the groups tested, a post hoc analysis revealed that no treatment was significantly different from the water control. Deer ked survival was different amongst the treatments, with deer keds exposed to permethrin dying much sooner than those exposed to other treatments or control (median survival for permethrin = 0.58 h). Post-hoc pairwise comparisons revealed that deer keds exposed to DEET or IR3535 had similar survival rates (4.82 and 5.15 h, respectively). Still, there were significantly lower survival times for DEET compared to OLE (6.33 h) and picaridin (15.00 h). Deer keds exposed to the water control survived the longest (23.12 h). Overall, deer ked repellency was not significantly different from the control, but permethrin-treated clothes can effectively kill deer keds in a short amount of time, thereby protecting those who recreate outdoors or encounter animals carrying deer keds., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The cosmopolitan ectoparasite human head louse, Pediculus humanus capitis (De Geer)(Phthiraptera:Pediculidae), affects mostly school-aged children, with infestations reported every year mainly due to louse resistance to pyrethroids. One of the main resistance mechanisms of pyrethroids is the target site insensitivity (kdr), which is caused by single-nucleotide point mutations (SNPs) located in the voltage-sensitive sodium channel gene. In this study, we analyzed individual head lice toxicologically via the description of their susceptibility profile to permethrin and genetically through the genotypification of their kdr alleles as well as nuclear microsatellite loci. Lice were collected from 4 schools in the city of Buenos Aires, Argentina. The resistance ratios varied from 33.3% to 71.4%, with a frequency of the T917I kdr mutation of 87.31% and with 83.6% of the head lice being homozygous resistant to pyrethroids. Microsatellite data indicated that all the louse school populations had genotype proportions that deviated from Hardy-Weinberg expectations, with FIS > 0 reflecting a deficit of heterozygotes. Bottleneck analysis suggested that all louse school populations underwent a recent reduction in population sizes, while 3 of the 4 schools had gene flow values around 1, indicating ongoing gene flow among those schools. Our study suggests that school louse populations in the city of Buenos Aires may form a metapopulation, where each school represents a small population that undergoes extinction and recolonization processes under strong permethrin selection. This is the first multilevel analysis integrating toxicological, kdr-genotyping, and microsatellite data in human louse populations., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

As observed in many locations worldwide, resistance to pyrethroids is common in Aedes aegypti (L.) in the southern United States and northern Mexico. Strong resistance in Aedes albopictus (Skuse) is less common and is not as well characterized. These 2 species have been undergoing range expansion and are sympatric in many locations including Houston, Texas. They are often collected from the same locations and lay eggs in the same larval habitats. In this study, we colonized both Ae. aegypti and Ae. albopictus from 4 locations in Houston and characterized insecticide resistance using permethrin as a model pyrethroid. We found differences in resistance intensity between the species at all 4 sites. Within the Ae. aegypti, resistance ratios ranged from 3.5- to 30.0-fold when compared to the ORL1952 laboratory susceptible strain. Expression of several P450s was higher than in the ORL1952 strain, but the pattern was similar between the field strains of Ae. aegypti. Higher resistance ratios did correlate with increasing percentages of the dilocus knockdown resistance (kdr) genotype. In contrast, Ae. albopictus from the 4 locations all had very low resistance ratios (<4-fold) when compared to the same laboratory susceptible strain. Five years later, we performed additional collections and characterization from the most resistant location to assess the temporal persistence of this difference in resistance between the species. The same pattern of high resistance in Ae. aegypti and low resistance in sympatric Ae. albopictus remained 5 yr later and this may have implications for operational efficacy., (Published by Oxford University Press on behalf of Entomological Society of America 2023.)

Rhipicephalus sanguineus s.l. (Latreille), is a vector of multiple disease-causing pathogens to humans and dogs. Permethrin and fipronil are two acaricides used to manage R. sanguineus s.l. infestations. Homeowners have reported treatment failures in managing brown dog ticks using permethrin and fipronil based products. Previous studies demonstrated that high permethrin resistance in some R. sanguineus s.l. populations was due to metabolic detoxification and target site insensitivity. In this study, three R. sanguineus s.l. strains, one from a laboratory colony (NC) and two colonies originally collected from Florida (FL) and California (CA), were evaluated for resistance expression against permethrin and fipronil. Metabolic detoxification mechanisms were evaluated in the FL strain using three synergists, while a polymerase chain reaction assay was used to detect a resistance mutation in all strains. The NC strain was susceptible to both permethrin and fipronil, while both the FL and CA strains exhibited high resistance to permethrin and tolerance to fipronil. The synergist tests and PCR results indicated that the FL strain utilized both metabolic resistance and target site insensitivity against permethrin, while the CA strain was documented to have the target-site insensitivity resistant allele. This study confirmed permethrin resistance in both California and Florida populations and its persistence in Florida populations, although its susceptibility can potentially be increased by adding a synergist. Fipronil resistance was not detected suggesting this acaricide may provide suitable tick control., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America.)

Pyrethroid resistance is widespread in sub-Saharan Africa. The objective of this study was to assess the insecticide resistance intensity in Anopheles gambiae s.l. (Diptera: Culicidae) in four districts of Benin in order to better understand how pyrethroid-only nets are likely to be effective. Thus, adult females of An. gambiae s.l., reared from field-collected larvae were used for assessing resistance intensity to permethrin and deltamethrin. They were tested at 1×, 5×, and 10× the diagnostic dose, using both WHO susceptibility tube testing and CDC bottle bioassays. Identification of molecular species, as well as of L1014F Kdr and Ace-1R mutations was performed using the PCR. The level of expression of biochemical enzymes was also evaluated. Overall, moderate to high resistance intensity to permethrin and deltamethrin was observed, irrespective of the testing method. While the L1014F Kdr frequency was high (>75%), Ace-1R was low (≤6%) in An. gambiaes.s. and Anopheles coluzzii, the two predominant species [52% (95% CI: 44.8-59.1) and 45% (95% CI: 38.0-52.2), respectively]. Anopheles arabiensis was found at very low frequency (3%, 95%CI: 1.1-6.4). For Biochemical analyses, α and β-esterases were over-expressed in all four districts, while mixed-function oxidases (MFOs) were over-expressed in only one. Overall, the two testing methods led to comparable conclusions, though there were a few inconsistencies between them. The moderate-high resistance intensity observed in the study area suggests that dual active-ingredient (AI) long-lasting insecticidal nets (LLINs) may provide better control of insecticide-resistant mosquitoes., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Amblyomma americanum Linnaeus (Ixodida: Ixodidae) is ubiquitously present throughout the southeastern United States and is capable of vectoring several pathogens. White-tailed deer are the main host for adult A. americanum. However, this tick species is a generalist that will feed on most vertebrates, including humans, deer, livestock, and pets. Management of this species can be challenging due to a lack of cost-effective strategies. Acaricides are often utilized, however, this may lead to pesticide resistance. The Food and Agriculture Organization of the United Nations (FAO) larval packet test (LPT) was performed on susceptible A. americanum to determine the lethal concentration (LC) and discriminating concentration (DC) values for permethrin. The FAO LPT was used at these pre-established values to compare levels of resistance in ticks collected from a captive deer farm and wild areas representing high and low permethrin exposure settings, respectively. Resistance ratios (RR) calculated from the LC values for the ticks collected from farmed and wild deer ranged between 1 and 2. A. americanum collected from farmed and wild deer were not found to be resistant, however, some samples had slightly elevated RRs as compared to the susceptible laboratory strain, which may suggest tolerance development. Although the A. americanum sampled in this study were not resistant to permethrin, the DC calculated in this study will allow for rapid evaluation of resistance in a permethrin resistance monitoring program such that alternate management strategies can be adopted if resistance is detected., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America.)

Long lasting permethrin-impregnated (LLPI) clothing can retain permethrin and repel ticks for up to three months and without exceeding EPA-approved safe levels; however, little is known about longer term effects of wearing LLPI clothing. Here, permethrin content was measured in new forester pants soon after initial impregnation (Insect Shield) and again one year later after being repeatedly worn by foresters in the field. Urine samples were collected from foresters for biomonitoring of permethrin metabolites at multiple time intervals (pre-use, one-month, three-to-four-months, and one-year post-use). Lethality against nymphal Ixodes scapularis Say was measured in clothing after one year of wear by foresters. Furthermore, to test potential variability in permethrin impregnation of different batches of clothing, separate sets of clothing were anonymously sent to Insect Shield for permethrin treatment over a period of three months and permethrin was quantified. Results demonstrated 33% of participants' pants had no measurable permethrin after one year of wear and permethrin content and tick mortality varied significantly between clothing. Only two of the participants' clothing resulted in ≥ 30% tick mortality after one year of wear. Significant differences were observed in 3-PBA and trans-DCCA, but not cis-DCCA metabolites in participants over the four measured time points and were higher than general United States population levels. This study provides practical information on the safety (measured by urinary metabolites) over time of LLPI clothing. It also provides snapshots (pre-washing and after one year of wear) of effectiveness of LLPI clothing as personal protective equipment against ticks for outdoor workers., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The development of insecticide resistance in different species of mosquitoes to Pyrethroids is a major challenge for vector-borne diseases transmitted by mosquitoes. Failure of Pyrethroids in control of mosquitoes would impact negatively on the gains recorded in control of mosquito-borne diseases in previous years. In anticipation of a country-wide deployment of Pyrethroid-treated nets for control of mosquito-borne diseases in Nigeria, this study assessed susceptibility of Culex quinquefasciatus Say. (Diptera: Culicidae) to Pyrethroids in Owhelogbo, Ejeme and Oria-Abraka communities in Delta State, Niger-Delta, Nigeria. Three to five day old Cx. quinquefasciatus were exposed to Deltamethrin (0.05%), Permethrin (0.75%), and Alphacypermethrin (0.05%) using World Health Organization bioassay method. Polymerase chain reaction (PCR) was employed in characterization of species and knockdown mutation. Results revealed that Cx. quinquefasciatus were generally susceptible (98-100%) to Deltamethrin, Permethrin, and Alphacypermethrin in the three communities with the exception of Owhelogbo where resistance to Deltamethrin (97%) was suspected. Knockdown time to Deltamethrin (11.51, 11.23, and 12.68 min), Permethrin (28.75, 13.26, and 14.49 min), and Alphacypermethrin (15.07, 12.50, and 13.03 min) were considerably low for Owhelogbo, Ejeme, and Oria-Abraka Cx. quinquefasciatus populations, respectively. Species identification result showed that all amplified samples were Cx. quinquefasciatus; however, no kdr allele was found in the three populations. Deployment of pyrethroid-treated nets for control of mosquito-borne diseases in Niger-Delta region of Nigeria is capable of reducing burden of diseases transmitted by Cx. quinquefasciatus as well as addressing nuisance value of the vector; however, caution must be entertained so as not to increase selection pressure thereby aiding resistance development., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The present study investigated in 8 villages of the Plateau region the coverage, usage, physical integrity, and bio-efficacy of the Olyset nets distributed nationwide by the Benin's National Malaria Control Programme in July 2011. The questionnaire administered as well as the observations made in the households allowed estimating the coverage and usage rates of the 2011 Olyset nets. While their physical integrity was assessed through standard WHO methodology, their bio-efficacy was evaluated through gas chromatography, and WHO cone testing performed with the Kisumu susceptible strain. Mosquito collections through human landing catches (HLCs) were also performed in torn nets to assess if a loss of protection of sleepers occurred as the nets fabric integrity got more damaged. Nine months postdistribution, the coverage and usage rates of the 2011 Olyset nets were 67.4% (95% CI: 65.8-68.9) and 73.3% (95% CI: 70.7-75.8) respectively. About 28% of the 2011 Olyset nets were torn. A drastic drop of the insecticide quantity on the fibers of the nets [from 7.08 µg (95% CI: 5.74-8.42) to 0.2 µg (95% CI: 0.01-0.38)] as well as mortality rates <80% were observed with most nets evaluated. Moreover, the biting rates of An. gambiae s.l. (Diptera: Culicidae) inside torn nets increased in line with their fabric integrity loss. These data support the conclusion that future deployment of nets in the field must be strengthened by community sensitization on their correct use in order to postpone as much as possible appearance of holes and loss of insecticidal activity and encourage repairing of torn nets., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Mass-trapping has been used to control outbreaks of Aedes aegypti (Linnaeus) (Diptera: Culicidae) in Puerto Rico since 2011. We investigated the effect of multi-year, insecticide-free mass trapping had on the insecticide susceptibility profile of Ae. aegypti. Eggs collected in southern Puerto Rico were used to generate F1 populations that were tested for susceptibility to permethrin, sumethrin, bifenthrin, deltamethrin, and malathion according to CDC bottle bioassays protocols. All populations of Ae. aegypti were resistant to the synthetic pyrethroids and mosquitoes from two locations were partially resistant to malathion. Population genetic analysis, using a double digest restriction sites associated DNA sequencing (ddRADseq) approach, indicated a large amount of migration between study sites effectively homogenizing the mosquito populations. Mass-trapping using noninsecticidal autocidal gravid ovitraps did not restore susceptibility to five active ingredients that are found in commercial insecticides. Migration between communities was high and would have brought outside alleles, including resistant alleles to the treatment communities. Further investigation suggests that household use of commercially available insecticide products may continue to select for resistance in absence of public health space spraying of insecticides., (Published by Oxford University Press on behalf of Entomological Society of America 2021.)

Pesticide resistance in medically significant disease vectors can negatively impact the efficacy of control efforts. Resistance research on ticks has focused primarily on species of veterinary significance that experience relatively high degrees of control pressure. Resistance in tick vectors of medical significance has received little attention, in part because area-wide pesticide applications are not used to control these generalist tick species. One of the few effective methods currently used for area-wide control of medically important ticks, including Ixodes scapularis Say (Acari: Ixodidae), is deployment of 4-poster devices. Deer self-apply a topical acaricide (permethrin) while feeding on corn from the devices. A 4-poster program using permethrin has been deployed on Shelter Island, NY to control I. scapularis populations since 2008. We collected engorged female ticks from deer in this management area and a location in the Mid-Hudson River Valley, NY without area-wide tick control. Larvae were reared from egg masses and their susceptibility to permethrin was tested. Larvae originating from a long-term laboratory colony were used as a susceptible baseline for comparison. Compared against the laboratory colony, resistance ratios at LC-50 for Shelter Island and Hudson Valley I. scapularis were 1.87 and 1.51, respectively. The susceptibilities of the field populations to permethrin were significantly lower than that of the colony ticks. We provide the first data using the larval packet test to establish baseline susceptibility for I. scapularis to permethrin along with information relevant to understanding resistance emergence in tick populations under sustained control pressure from 4-poster devices., (Published by Oxford University Press on behalf of Entomological Society of America 2021.)

Highly residual pyrethroids such as permethrin have been used for controlling mosquitoes that transmit infectious diseases. However, the selective pressure from such insecticides may result in cross-resistance against other pyrethroids used for household insecticides. In this study, we investigated the susceptibility of Culex quinquefasciatus Say collected from Brazil and Myanmar to permethrin in addition to four types of household pyrethroids. Both strains exhibited high resistance against all pyrethroids tested, indicating cross-resistance. Furthermore, we detected the knockdown resistance (kdr) mutations L932F+I936V in the voltage-gated sodium channel gene (VGSC) in the Brazilian strain. Notably, the L932F+I936V haplotype has previously been observed in in silico data, but it should be detected not directly from living insects. In comparison, a common kdr mutation, L1014F, was detected from the Myanmar strain. Although L1014F was also detected from the Brazilian strain, the allele frequency was too low to affect resistance. Both strains harbored the resistance-associated haplotypes of the cytochrome P450 gene, CYP9M10. The Brazilian strain demonstrated comparable resistance against pyrethroids as that of the Myanmar strain even when a cytochrome P450 inhibitor, piperonyl butoxide was added to the bioassay. Our results suggested that the L932F+I936V mutations confer the Brazilian strain of Cx. Quiquefasciatus with resistance at a comparable level to that conferred by the well-recognized kdr mutation L1014F in the Myanmar strain. The identification of unexplored mutations may improve the diagnosis and understanding of resistance of this medically important species., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

There are major public health concerns regarding the spread of mosquito-borne diseases such as dengue, Zika, and chikungunya, which are mainly controlled by using insecticides against the vectors, Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse). Pyrethroids are the primary class of insecticides used for vector control, due to their rapid knockdown effect and low toxicity to vertebrates. Unfortunately, continued use of pyrethroids has led to widespread insecticide resistance in Ae. aegypti; however, we lack information for Ae. albopictus-a sympatric species in Chiapas since 2002. In this study, we evaluated the permethrin resistance status of Ae. albopictus collected from Mexico and Texas. We also selected for permethrin resistance in the laboratory and investigated the potential mechanisms conferring resistance in this species. Knockdown resistance mutations, specifically F1534C, in the voltage-gated sodium channel gene, and increased activity of detoxifying enzymes were evaluated. Low levels of permethrin resistance (<2.4-fold) were observed in our field populations of Ae. albopictus and the F1534C mutation was not detected in any of the sites. Low levels of resistance were also observed in the artificially selected strain. There was significantly higher cytochrome P450 activity in our permethrin-selected and nonselected strains from Mexico compared to the control strain. Our results suggest the Ae. albopictus sampled from 2016 are mostly susceptible to pyrethroids. These results contrast with the high levels of permethrin resistance (>58-fold) found in Ae. aegypti from the same sites in Mexico. This research indicates the importance of continued monitoring of Ae. albopictus populations to prevent resistance from developing in the future., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Aedes aegypti (L.) is the primary vector of Zika, dengue, yellow fever, and chikungunya viruses. Insecticides used in mosquito control can help prevent the spread of vector-borne diseases. However, it is essential to determine insecticide resistance (IR) status before control measures are undertaken. Only the most effective insecticides should be used to avoid ineffective control and/or promotion of IR. Pyrethroids and organophosphates are the most commonly used insecticides for mosquito control. Here, the efficacy of two active ingredients (AIs; permethrin [pyrethroid], chlorpyrifos [organophosphate]), two formulated products (FPs; Biomist [AI: permethrin]) and (Mosquitomist [AI: chlorpyrifos]), and three synergists (piperonyl butoxide, diethyl maleate, S-S-S-tributyl phosphorotrithioate) was evaluated in two Ae. aegypti colonies (pyrethroid resistant and susceptible). Mosquitomist was most effective against the pyrethroid-resistant colony (100% mortality at diagnostic time). Pre-exposure to synergists did not increase the efficacy of AIs against the pyrethroid-resistant colony. Further research is needed to discover how synergists may affect the efficacy of insecticides when used on pyrethroid-resistant mosquitoes., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Aedes aegypti (Linnaeus, 1762) is considered the most important mosquito vector species for several arboviruses (e.g., dengue, chikungunya, Zika) in Costa Rica. The primary strategy for the control and prevention of Aedes-borne diseases relies on insecticide-based vector control. However, the emergence of insecticide resistance in the mosquito populations presents a significant threat to these prevention actions. The characterization of the mechanisms driving the insecticide resistance in Ae. aegypti is vital for decision making in vector control programs. Therefore, we analyzed the voltage-gated sodium channel (VGSC) gene for the presence of the V1016I and F1534C kdr mutations in Ae. aegypti populations from Puntarenas and Limon provinces, Costa Rica. The CDC bottle bioassays showed that both Costa Rican Ae. aegypti populations were resistant to permethrin and deltamethrin. In the case of kdr genotyping, results revealed the co-occurrence of V1016I and F1534C mutations in permethrin and deltamethrin-resistant populations, as well as the fixation of the 1534C allele. A strong association between these mutations and permethrin and deltamethrin resistance was found in Puntarenas. Limon did not show this association; however, our results indicate that the Limon population analyzed is not under the same selective pressure as Puntarenas for the VGSC gene. Therefore, our findings make an urgent call to expand the knowledge about the insecticide resistance status and mechanisms in the Costa Rican populations of Ae. aegypti, which must be a priority to develop an effective resistance management plan., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The use of insecticides such as deltamethrin, permethrin, and temephos has been a primary tool to manage Aedes aegypti (Linnaeus) in Punjab province, Pakistan; however, recent reports of control failure necessitate monitoring insecticide resistance. For this reason, we evaluated 12 field strains of Ae. aegypti from Punjab for resistance against deltamethrin, permethrin, and temephos along with underlying resistance mechanisms. For deltamethrin, high level of resistance was observed in Rawalpindi, Faisalabad, Sheikhupura, Lahore, Pattoki, and Kasur strains (RRLC50 > 10-fold); moderate level of resistance in Sargodha, Gujranwala, and Sialkot strains (RRLC50 = 5- to 10-fold), and low level of resistance in Okara, Multan, and Sahiwal strains (RRLC50 < 5-fold). In the case of permethrin, high level of resistance was found in all the field strains, except the Okara strain that exhibited moderate resistance. For temephos, five field strains, viz. Faisalabad, Rawalpindi, Kasur, Lahore, and Gujranwala, showed high level of resistance; five strains, viz. Sheikhupura, Sialkot, Pattoki, Sahiwal, and Okara, showed moderate resistance, and two strains from Multan and Sargodha showed low resistance to temephos. Synergism bioassays implementing piperonyl butoxide and S,S,S-tributylphosphorotrithioate exhibited a nonsignificant effect on synergizing toxicity of deltamethrin and permethrin in all field strains except the Lahore strain, suggesting the possible role of target-site resistance mechanism. However, both synergists had a significant effect on synergizing toxicity of temephos in all field strains, suggesting the possibility of metabolic-based mechanism of insecticide resistance. In conclusion, the study confirmed the presence of resistance to deltamethrin, permethrin, and temephos in the studied field strains of Ae. aegypti from Punjab, Pakistan., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

G-protein-coupled receptors (GPCRs) are a large family of seven-transmembrane domain proteins that exist in plants and animals, playing critical physiological functions through intracellular cascades. Previous studies revealed an important regulation pathway of GPCR/Guanine nucleotide-binding protein (G-protein)/Adenylyl Cyclase (AC)/cAMP-dependent protein kinase A (PKA) in the insecticide resistance and regulation of resistance-related P450 gene expression in highly resistant southern house mosquitoes, Culex quinquefasciatus Say (Diptera: Culicidae). However, the function of this regulation pathway in field-collected and laboratory-susceptible mosquitoes is still unknown. In the current study, we characterized the function of each effector (GPCR, G-protein, AC, and PKA) in this GPCR intracellular pathway in both field-collected and laboratory Cx. quinquefasciatus strains, showing that knockdown of the expression of each effector gene can cause 1) decreased expression of their downstream respective genes and 2) increased sensitivity of the mosquitoes to permethrin insecticide. These results, together with our previous findings, strongly suggest the universal function of the GPCR-regulation cascade in the mosquito's sensitivity to insecticides and its regulation of resistance development through P450-mediated detoxification.

Although the head louse, Pediculus humanus capitis De Geer, and body louse, Pediculus humanus humanus L., both have a worldwide distribution, the occurrence of head louse pediculosis appears to be more prevalent in modern societies despite systematic use of various pediculicides. This study tested head lice collected in rural Georgia and body lice collected in Russia for the prevalence of a kdr-biomarker that is associated with permethrin resistance. This study also screened lice for the presence of DNA from Bartonella quintana and Acinetobacter species. The kdr-permethrin resistance biomarker for the T917I mutation was detected by RFLP and PCR in 99.9% of head lice tested from Georgia, whereas only 2.9% of body lice from Russia tested positive for this kdr biomarker. DNA of B. quintana was detected in 10.3% of head lice from Georgia, whereas 84.8% of body lice from Russia tested positive. Acinetobacter DNA was detected in 80.8% (95% CI, 68-89%) of head lice from Georgia and all body lice from Russia tested., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Permethrin resistance is widespread in Aedes aegypti (L.), the main dengue, zika, and chikungunya virus vector in Latin America and the Caribbean. A common mechanism of resistance to pyrethroids-knockdown resistance (kdr)-is conferred through mutations in the insect's voltage-dependent sodium channel. In this mosquito, around 10 replacement substitutions in the voltage-gated sodium channel gene (vgsc) have been reported in pyrethroid-resistant strains. Two of these mutations, named Ile1,016 and Cys1,534, are widespread in mosquito populations from Latin America and the Caribbean. This study assessed the levels of permethrin resistance and the frequency of two kdr mutations in eight Ae. aegypti populations collected in Puerto Rico in 2013. Permethrin resistance factors ranged from 33-214-fold relative to the New Orleans reference strain. The frequency of kdr mutation Ile1,016 ranged from 0.65 to fixation (1.0), and for Cys1,534 frequencies varied from 0.8 to fixation. Alarmingly, two populations-Carolina and Caguas-reached fixation at both loci. Our results suggest that permethrin effectiveness for Ae. aegypti control is compromised in these collections from Puerto Rico., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Phlebotomus papatasi is one of the most medically important sand fly species in the Old World, serving as a vector of Leishmania parasites and phleboviruses. Chemical control is still considered the most effective method for rapidly reducing populations of flying insects involved in vector-borne disease transmission, but is increasingly threatened by insecticide resistance in the target insect posing significant problems for entomologists responsible for control programs. This study was conducted to determine pyrethroid resistance mechanisms and the biological, physiological, and molecular impacts of resistance in Ph. papatasi, and to compare their resistance mechanisms against those reported for mosquitoes and other intensely studied dipterans. Field-collected Ph. papatasi from Aswan, Egypt, were subjected to sublethal doses of permethrin and reared as a resistant strain under laboratory conditions through 16 generations. Biological parameter observations of resistant Ph. papatasi revealed an association of resistance with productivity cost. Physiological analysis revealed that concentrations of oxidase and esterase enzymes increased in early generations of the resistant colony, and then subsided through the F16 generation to levels similar to those in a susceptible colony. The activity levels of acetylcholinesterase were higher in field-collected Ph. papatasi than in susceptible colony flies, but decreased significantly despite subsequent exposure to permethrin. The molecular search for gene mutations in the resistant strain of Ph. papatasi failed to identify any mutations common in pyrethroid-resistant mosquitoes. Our study revealed that the mechanism of pyrethroid resistance in sand flies is different than that in mosquitoes, at least at the genetic level., (Published by Oxford University Press on behalf of Entomological Society of America 2016. This work is written by US Government employees and is in the public domain in the US.)

A field strain of Aedes aegypti (L.) was collected from Puerto Rico in October 2008. Based on LD50 values by topical application, the Puerto Rico strain was 73-fold resistant to permethrin compared with a susceptible Orlando strain. In the presence of piperonyl butoxide, the resistance of Puerto Rico strain of Ae. aegypti was reduced to 15-fold, suggesting that cytochrome P450-mediated detoxification is involved in the resistance of the Puerto Rico strain to permethrin. To determine the cytochrome P450s that might play a role in the resistance to permethrin, the transcriptional levels of 164 cytochrome P450 genes in the Puerto Rico strain were compared with that in the Orlando strain. Of the 164 cytochrome P450s, 33 were significantly (P < 0.05) up-regulated, including cytochrome P450s in families four, six, and nine. Multiple studies have investigated the functionality of family six and nine cytochrome P450s, therefore, we focused on the up-regulated family 4 cytochrome P450s. To determine whether up-regulation of the four cytochrome P450s had any functional role in permethrin resistance, transgenic Drosophila melanogaster Meigen lines overexpressing the four family 4 P450 genes were generated, and their ability to survive exposure to permethrin was evaluated. When exposed to 5 microg per vial permethrin, transgenic D. melanogaster expressing CYP4D24, CYP4H29, CYP4J15v1, and CYP4H33 had a survival rate of 60.0 +/- 6.7, 29.0 +/- 4.4, 64.4 +/- 9.7, and 11.0 +/- 4.4%, respectively. However, none of the control flies survived the permethrin exposure at the same concentration. Similarly, none of the transgenic D. melanogaster expressing CYP4J15v1 or CYP4H33 ?5 survived when they were exposed to permethrin at 10 microg per vial. However, transgenic D. melanogaster expressing CYP4D24 and CYP4H29 had a survival rate of 37.8 +/- 4.4 and 2.2 +/- 2.2%, respectively. Taken together, our results suggest that CYP4D24 might play an important role in cytochrome P450-mediated resistance to permethrin.

ABSTRACT Phytochemicals have been considered as alternatives for conventional pesticides because of their low mammalian toxicity and environmental safety. They usually display less potent insecticidal effects than synthetic compounds, but may express as yet unknown modes of action. In the current study, we evaluated 14 plant essential oils for their toxicities and synergistic effects with carbaryl and permethrin against fourth instars of Aedes aegypti (L.) as well as 5-7-d-old adults. Six essential oils showed significant synergistic effects with carbaryl at 10-50 mg/liter, but paradoxically all of them decreased the toxicity of permethrin against Ae. aegypti larvae. None showed toxicity or synergistic effects on Ae. aegypti adults, at doses up to 2,000 ng/ insect. The six essential oils displaying synergistic effects in Ae. aegypti larvae inhibited the in vitro activities of cytochrome P450 monooxygenases and carboxylesterases in the low milligram per liter range. The data indicated that cytochrome P450 monooxygenases and carboxylesterase were probably targets for these natural synergists. Thus, the mechanism of synergism was most likely inhibition of metabolism and not interacting target site effects.

The primary malaria vector in sub-Saharan Africa, Anopheles gambiae Giles (Diptera: Culicidae), is an anthropophagic and endophilic mosquito targeted for control with insecticides applied to interior resting surfaces and impregnated onto bed net materials. Effective malaria vector management involves monitoring the insecticide susceptibility of mosquito populations. Contemporary bioassays are based on mosquito contact with insecticide residues. We developed an innovative insecticide bioassay system that involves mosquito ingestion of a sugar-insecticide solution. The sucrose-permethrin solution in our bioassay system contained Trypan blue dye, creating a visual marker of insecticide ingestion in the mosquito's abdomen. Blue fecal spots deposited in the bioassay container provided further evidence of mosquito feeding. We used our bioassay to characterize the permethrin susceptibility of adult females of two strains of A. gambiae, one of which was susceptible and the other exhibited reduced susceptibility to permethrin. We compared the dose-response of both strains to permethrin in a forced-contact filter paper bioassay. Both assay approaches produced similar dose-dependent mortality, indicating that the feeding bioassay had appropriately characterized permethrin susceptibility for both mosquito strains.

Insecticides, especially pyrethroids, are important components in the vector-control effort. To better understand the development of resistance, the current study characterized resistance profiles in individual single-egg-raft colonies of a field population of Culex quinquefasciatus Say, HAmCq(G0). Our study, with 104 colonies derived from each of the single-egg-rafts of HAmCq(G0), indicated that the levels of resistance to permethrin in fourth instar larvae ranged from 0.4- to 280-fold compared with laboratory susceptible S-Lab larvae. We characterized the distribution of single-egg-raft colonies with different levels of resistance in the HAmCq(G0) population and found that 65% individual colonies had < 10-fold levels of resistance to permethrin, 16% from 10- to 20- fold, 7% from 20- to 30-fold, and 12% < or = 30-fold. We further characterized the frequency of the L-to-F kdr allelic expression of sodium channels in the single-egg-raft colonies with different levels of resistance to determine its possible role in resistance. The correlation between allelic expression and levels of resistance clearly showed the importance of L-to-F kdr mutation mediated sodium channel insensitivity in resistance development. However, our results also suggested that the sodium channel insensitivity is unlikely to be the sole mechanism and multiple mechanisms may present among the single colonies in response to insecticide resistance.

Insecticide resistance has become a serious issue in vector management programs. Information on insecticidal resistance and its associated mechanisms is important for successful insecticide resistance management. The selection of a colony of permethrin-resistant Aedes albopictus (Skuse) (Diptera: Culicidae), originating from Penang Island, Malaysia, yielded high larval-specific resistance to permethrin and cross-resistance to deltamethrin. Synergism assays showed that the major mechanism underlying this resistance involves cytochrome P450 monooxygenase. The resistance is autosomal, polygenically inherited and incompletely dominant (D = 0.26). Resistant larvae were reared under different conditions to assess the fitness costs. Under high larval density, larval development time of the resistant SGI strain was significantly longer than the susceptible VCRU strain. In both high- and low-density conditions SGI showed a lower rate of emergence and survival compared with the VCRU strain. Resistant larvae were more susceptible to predation by Toxorhynchites splendens (Wiedemann) (Diptera: Culicidae) larvae. The body size of SGI females reared under high-density conditions was larger compared with females of the susceptible strain. SGI females survived longer when starved than did VCRU females. The energy reserve upon eclosion was positively correlated with the size of the adults.

We conducted field trials to compare the relative repellent activity of two natural product compounds (nootkatone and carvacrol) with commercially available plant-derived (EcoSMART organic insect repellent) and permethrin-based (Repel Permanone) repellents against adult Ixodes scapularis Say and Amblyomma americanum (L.) (Acari: Ixodidae) by using treated coveralls. One day after treatment, nootkatone and carvacrol provided 100% repellency of I. scapularis adults, with nootkatone maintaining complete protection through 3 d, whereas carvacrol showed steadily declining repellency against I. scapularis during the 7-d course of the trials. Nootkatone was at least as effective against host-seeking A. americanum as against I. scapularis through 3 d. Carvacrol provided little protection against A. americanum adults. Both natural compounds performed well initially in comparison with the commercial products. After 7 d, nootkatone was the most effective against both species followed in order of activity by Permanone, EcoSMART, and carvacrol. Nootkatone seems to have offer considerable potential as a clothing repellent against both I. scapularis and A. americanum.

The cDNA of a trypsin gene from Aedes (Ochlerotatus) taeniorhynchus (Weidemann) was cloned and sequenced. The full-length mRNA sequence (890 bp) for trypsin from Ae. taeniorhynchus (AetTryp1) was obtained, which encodes an open reading frame of 765 bp (i.e., 255 amino acids). To detect whether AetTryp is developmentally regulated, a quantitative real-time polymerase chain reaction was used to examine AetTrypl mRNA expression levels in different developmental stages of Ae. taeniorhynchus. AetTryp1 was expressed at low levels in egg, larval, and pupal stages, but was differentially expressed in adult Ae. taeniorhynchus, with highest levels found in 5-d-old female adults when compared with teneral adults. In addition, AetTryp1 mRNA expression differed between sexes, with expression levels much lower in males. However, in both males and females, there was a significant increase in AetTryp1 transcription levels as age increased and peaked in 5-d-old adults. AetTrypl expressed in 5-d-old female Ae. taeniorhynchus significantly increased after 30 min postblood feeding compared with the control. The AetTryp1 mRNA expression in 5-d-old female Ae. taeniorhynchus was affected by different concentrations of permethrin.

The number of tick bites received by individuals wearing either permethrin-treated or untreated summer clothing (T-shirt, shorts, socks, and sneakers) was compared during a controlled indoor study. Pathogen-free nymphal Ixodes scapularis Say were placed on the left shoe, right leg, and left arm of 15 (5/treatment group/d) human volunteers wearing untreated outfits or outfits treated with permethrin either commercially or using a do-at-home treatment kit. The number and location of ticks attached to subjects' skin were recorded 2.5 h postinfestation. Subjects wearing outfits treated with permethrin received 3.36 times fewer tick bites than subjects wearing untreated outfits. No statistically significant differences in number of tick bites were detected between commercial permethrin treatment (19.33%) and the do-at-home permethrin application method (24.67%). The success of permethrin-treated clothing in reducing tick bites varied depending on the specific article of clothing. Subjects wearing permethrin-treated sneakers and socks were 73.6 times less likely to have a tick bite than subjects wearing untreated footware. Subjects wearing permethrin-treated shorts and T-shirts were 4.74 and 2.17 times, respectively, less likely to receive a tick bite in areas related to those specific garments than subjects wearing untreated shorts and T-shirts. Ticks attached to subjects were classified as alive or dead before removal. On subjects wearing untreated outfits, 97.6% of attached nymphs were alive, whereas significantly fewer (22.6%) attached nymphs were alive on subjects wearing repellent-treated outfits. Results of this study demonstrate the potential of permethrin-treated summer clothing for significantly reducing tick bites and tick-borne pathogen transmission.

The toxicity of permethrin to different life stages of Culex quinquefasciatus (Say) was examined in three field insecticide resistant strains, HAmCq(G0), MAmCq(G0), and BAmCq(G0); two permethrin selected strains, HAmCq(G8) and MAmCq(G6); and a laboratory susceptible strain, S-Lab. The levels of tolerance to permethrin differed among the developmental stages of Culex mosquito larvae with an ascending order from first to fourth instar. A developmentally regulated pattern of permethrin resistance in Culex mosquito larvae was identified, once again increasing from first to fourth instar. Adult mosquitoes had relatively lower levels of resistance in general compared with their larval stages, suggesting that different mechanisms may be involved in the response to insecticide selection pressure between larvae and adults. Inheritance of resistance to permethrin was examined using reciprocal crosses between resistant HAmCq(G8) and susceptible S-Lab strains; the resulting logarithm concentration-probit response curves for reciprocal F1 lines were similar to those for the susceptible strain, with degree of dominance values of -0.2 to -0.3, suggesting that the inheritance of permethrin resistance in Cx. quinquefasciatus is incompletely recessive. No significant difference was found in the values of LC50 and the slope of the log concentration-probit lines between reciprocal F1 lines, indicating that the resistance to permethrin was inherited autosomally. Both chi2 goodness-of-fit test and log concentration-probit line analysis on responses of backcross and self-bred progeny to permethrin indicated that permethrin resistance in the HAmCq(G8) strain did not follow a monogenic gene inheritance model, revealing that permethrin resistance in Cx. quinquefasciatus is controlled by more than one gene.

Insecticide resistance in populations of mosquitoes is an escalating problem that can directly affect disease prevalence. Determining the fitness associated with an insecticide resistance mechanism (allele) will provide for greater understanding of the evolution of resistance, and help inform effective vector management programs. Previously, a population cage experiment in which the alleles of two highly related strains of Culex pipiens quinquefasciatus (Say) SLAB (susceptible) and ISOP450 (permethrin resistant because of cytochrome P450-mediated detoxification) were placed in direct competition in the absence of insecticide revealed that the P450 resistance allele frequency decreased over time. In the current study, SLAB and ISOP450 development, mortality and energetic resources derived from glycogen, other sugars and lipids were measured to identify biological parameters that might explain the previously observed fitness cost. SLAB exhibited shorter egg-to-adult female development time and larger body size when reared in groups when compared with ISOP450. ISOP450 female adults provided 20% sugar water lived longer than 20% sugar water fed females of the SLAB strain. No significant differences in larval development time, larval mortality, pupal stage duration time, pupal mortality, longevity of male and female adults provided with distilled water and males provided sugar water were found between the strains. The caloric content from glycogen and lipids were significantly higher in SLAB relative to ISOP450 in adults. The slower female emergence time and smaller body size when reared in groups combined with lower energy reserves (glycogen and lipids) associated with the resistance allele (in ISOP450) are likely fitness costs associated with the resistance allele of P450-mediated detoxification.

Cytochrome b, coded by mitochondrial DNA, is one of the cytochromes involved in electron transport in the respiratory chain of mitochondria. Cytochrome b is a critical intermediate in a mitochondrial death pathway. To reveal whether cytochrome b of the mosquito Aedes aegypti (L.) (Diptera: Culicidae) (AeaCytB) is developmentally regulated, we used real-time quantitative polymerase chain reaction (qPCR) to examine AeaCytB gene expression levels in different developmental stages of Ae. aegypti. The qPCR showed that AeaCytB was expressed in each developmental stage, with peaks at first and second instars and was highly expressed in teneral male and female Ae. aegypti adults. Because mitochondrial genes exist as multiple copies, AeaCytB has much higher expression levels in all developmental stages in Ae. aegypti compared with nuclear genes. We also investigated the effect of abiotic environmental factors (e.g., high temperatures, ultraviolet radiation, and pesticide) on AeaCytB gene expression. Taken together, these results suggest that AeaCytB gene plays an important role in the development of Ae. aegypti and its response to environmental stress.

Using quantitative polymerase chain reaction (QPCR), the relative transcriptional levels of cytochrome c oxidase subunit 3 (CO3) were studied in Aedes aegypti in response to treatment with acetone, permethrin, and fipronil. The transcriptional levels of CO3 were significantly (P < 0.05) higher in acetone-treated Ae. aegypti compared with that in untreated samples. Using ribosomal L24, heat shock protein (HSP), and actin as reference genes, relative transcription levels of CO3 in acetone-treated Ae. aegypti were 2.88 +/- 0.38-, 2.60 +/- 0.60-, and 3.24 +/- 0.70-fold higher, respectively, compared with that in untreated mosquitoes. Transcriptional levels of CO3 were induced significantly higher (6.54 +/- 1.22-, 4.62 +/- 0.74-, and 9.47 +/- 3.71-fold, respectively) by permethrin at LD10 compared with acetone (P < 0.05). Taken together, our results suggest that overexpression of CO3 is tightly regulated in Ae. aegypti in response to xenobiotic treatment.

Using the polymerase chain reaction (PCR)-select subtractive cDNA hybridization technique, 18 different genes were isolated from a permethrin-treated versus acetone-treated Aedes aegypti subtractive library. Quantitative PCR (QPCR) results showed that 8 of the 18 gene's transcriptional levels in permethrin-treated Ae. aegypti were at least two-fold higher (ranging from 2.6 +/- 0.5 to 4.8 +/- 0.2) than that in acetone-treated Ae. aegypti. These eight genes include three functionally known genes (cytochrome c oxidase subunit III, NADH2 dehydrogenase, deltamethrin resistance associated protein), three functionally unknown genes (Ae. aegypti putative 16.9-kDa secreted protein, Anopheles gambiae ENSANGP00000019508, Cryptococcus neoformans hypothetical protein CNE05340), and two novel genes. Transcriptional levels for 11 of the 18 genes were induced significantly higher by permethrin than by fipronil (P < 0.05). Our results suggest that subtractive cDNA hybridization and QPCR are powerful techniques to identify differentially expressed genes in response to pesticide treatment.

Apoptosis (programmed cell death) is a tightly regulated physiological process. The inhibitors of apoptosis proteins (IAPs) are key regulators for apoptosis. An inhibitor of apoptosis protein gene IAP1 was recently cloned from Aedes aegypti (L.) (AaeIAP1, GenBank accession no. DQ993355); however, it is not clear whether AaeIAP1 is developmentally and environmentally regulated. In this study, we applied quantitative polymerase chain reaction (PCR) to investigate the expression levels of the AaeIAPI transcript in different developmental stages and under different environmental conditions. Our results revealed that the expression of the AaeIAP1 transcript was detectable in all life stages ofAe. aegypti, with significantly higher levels in pupal and adult stages than in larval stages. Furthermore, when Ae. aegypti was exposed to all stressful environmental conditions (e.g., low and high temperatures, UV radiation, acetone, and permethrin insecticide treatment), the expression level of AaeIAP1 transcript was increased significantly. Our results suggest that AaeIAP1 might play an important role in both the physiological development ofAe. aegypti and stress-induced apoptosis.

A quantitative sequencing (QS) protocol that detects the frequencies of sodium channel mutations (M815I, T917I, and L920F) responsible for knockdown resistance in permethrin-resistant head lice (Pediculus humanus capitis De Geer) was tested as a population genotyping method for use as a preliminary resistance monitoring tool. Genomic DNA fragments of the sodium channel a-subunit gene that encompass the three mutation sites were polymerase chain reaction (PCR)-1 amplified from individual head lice with either resistant or susceptible genotypes, and combined in various ratios to generate standard DNA template mixtures for QS. After sequencing, the signal ratios between resistant and susceptible nucleotides were calculated and plotted against the corresponding resistance allele frequencies. Quadratic regression coefficients of the plots were close to 1, demonstrating that the signal ratios are highly correlated with the resistance allele frequencies. Resistance allele frequencies predicted by QS, using either "pooled DNA" (DNA extracted from individual louse specimens and pooled) or "pooled specimen DNA" (DNA simultaneously extracted from multiple louse specimens), agreed well with those determined by individual sequencing, confirming the reliability and accuracy of QS as a population genotyping method and validating our approach of using the pooled specimen DNA as the DNA template for QS. Our protocol for QS was determined to be highly reliable for the prediction of resistance allele frequencies higher than approximately 7.4% at the 95% confidence level. According to the resistance allele frequencies determined by QS, pyrethroid resistance varies substantially among different geographical regions, emphasizing the importance of early resistance detection and proper management strategies.

The Santa Luiza strain of the southern cattle tick, Boophilus microplus (Canestrini) (Acari: Ixodidae), is resistant to both permethrin and amitraz. A study was conducted at the USDA Cattle Fever Tick Research Laboratory in Texas to investigate the genetic basis of permethrin resistance with cross-mating experiments, and to determine the mechanisms of permethrin resistance through synergist bioassays and biochemical analysis of esterase profiles. The Muñoz strain, an acaricide-susceptible reference strain, was used as the susceptible parent and the Santa Luiza strain, originating in Brazil, was used as the resistant parent. The Food and Agriculture Organization larval packet test was used to measure the levels of susceptibility of larvae of the parental strains, F1, backcross, F2, and F3 generations to permethrin. Results of reciprocal crossing experiments suggested that permethrin resistance was inherited as an incomplete recessive trait. There was no significant maternal effect on larval progeny's susceptibility to permethrin in the F1 and subsequent generations. The values of the degree of dominance were estimated at -0.700 and -0.522 for the F1 larvae with resistant and susceptible female parents, respectively. Results of bioassays on larval progeny of the F1 backcrossed with the resistant parent strain and of the F2 generations suggested that one major gene was responsible for permethrin resistance in the Santa Luiza strain. Selection of F3 larvae with either permethrin or amitraz led to significantly increased resistance to both permethrin and amitraz, indicating a close linkage between genes responsible for permethrin and amitraz resistance. The possible involvement of metabolic enzymes in permethrin resistance in the Santa Luiza strain of B. microplus was dismissed by the lack of enhanced synergism by TPP or PBO, as observed in synergist bioassays, as well as by the lack of enhanced esterase activity in the Santa Luiza strain relative to the susceptible strain. The results of this study suggest that other mechanisms, including a possible new sodium channel mutation that is different from the one currently known, may be responsible for permethrin resistance in the Santa Luiza strain of B. microplus.

This study examines the effectiveness of a new ivermectin formulation for the topical treatment of the human head louse, Pediculus humanus capitis De Geer (Anoplura: Pediculidae). Permethrin-resistant lice originally obtained from south Florida and maintained on an in vitro rearing system were 100% susceptible to ivermectin formulations by using a semiclinical hair tuft bioassay. The formulation was 100% effective at killing lice using 1, 0.5, and 0.25% ivermectin concentrations after 10-min exposures. As judged by the lethal time (LT)50 and LT95 values, 0.5% formulated ivermectin was 3.8 and 3.2 times faster at killing lice, respectively, than 0.5% nonformulated ivermectin, indicating that the formulation may facilitate the penetration of ivermectin into the louse. The hair tuft-based bioassay in conjunction with the in vitro rearing system provides a standardized method to assess the comparative efficacy of pediculicide formulations in a reproducible format that mimics the exposure scenario that occurs on the human scalp.

The effects of different pyrethroid insecticides, formulations, and doses on peridomestic populations of Triatoma infestans (Klug) were evaluated in 128 houses with 148 identified infested peridomestic sites in northwestern Argentina between October 2003 and March 2005. Four treatments were randomly assigned within each community: two doses of 5% suspension concentrate beta-cypermethrin in water applied with manual compression sprayers, the standard dose (S) at 50 mg and a double dose (2S) at 100 mg active ingredient (AI)/m2; and two emulsifiable concentrates diluted in diesel fuel and applied with power sprayers, 25% cypermethrin (100 mg [AI] /m2) (CF) and 10% permethrin (170 mg [AI]/m2) (DF). Infestation was assessed by timed manual collections with a dislodging agent at baseline, 5, 12, and 17 mo postspraying, and the sites found to be reinfested at 5 mo postspraying were selectively resprayed. Only 2S eliminated T. infestans from all peridomestic sites up to 12 mo postspraying, and it was significantly more effective than all other treatments. At 5 mo postspraying, more sites treated with CF or DF rather than S had bug colonies that probably represented residual foci, which they also failed in eliminating after a second spray. At 17 mo postspraying, the prevalence of reinfested peridomestic sites was 5% for 2S, 29% for S, 43% for CF, and 54% for DF. The application of suspension concentrate pyrethroids in dose twice as large as that currently in use in the attack phase produces a greater initial impact and may eliminate peridomestic populations of T. infestans.

The horn fly, Haematobia irritans irritans (L.) (Diptera: Muscidae), has become a problem for Brazilian cattle producers even though its introduction into Brazil is relatively recent. Failure to control this cattle pest is becoming a concern, and horn fly populations from several ranches from the state of Mato Grosso do Sul were surveyed for pyrethroid resistance. Susceptibility bioassays revealed that cypermethrin resistance was widespread and reached high levels in horn fly populations throughout the state, with resistance factors (RFs) ranging from 50.4 to 704.8. Synergist bioassays failed to detect a major role for esterases as a pyrethroid resistance mechanism in these populations, except for the highly pyrethroid-resistant Estrela do Oeste population (RF = 704.8). The kdr sodium channel gene mutation was not detected in eight of the 13 populations, but < 7% of individuals from four populations and 50% of the flies from Estrela do Oeste exhibited this mutation. Neither the superkdr sodium channel gene mutation nor a resistance-associated gene mutation in the HialphaE7 carboxylesterase were found in any of the fly populations. Although target site insensitivity (kdr) and esterase-mediated metabolism occur in horn fly populations from Mato Grosso do Sul state, it seems that they are not the major mechanism causing pyrethroid resistance in most of these populations.

Insecticide-treated bednets (ITNs) significantly reduce malaria vector populations. Susceptibility to ITNs differs by vector species, and culicine mosquitoes have not been shown to be significantly affected by the use of ITNs. We examined the impact of 2-4 yr of ITN use on malaria vector species distribution and culicine mosquitoes. Routine entomological surveillance was conducted in adjacent areas with and without ITNs from November 1999 to January 2002. Use of ITNs reduced the proportion of Anopheles gambiae Giles relative to Anopheles arabiensis Giles. The number of culicines per house was significantly lower in the ITN area than in the neighboring area. Changes in the An. gambiae sibling species distribution may help to explain apparent mosquito behavioral changes attributed to ITNs. Reductions in culicines by ITNs may have implications for community perceptions of ITN effectiveness and for control of other diseases such as lymphatic filariasis.

Eleven cat flea, Ctenocephalides felis (Bouchè), strains, seven field-collected and four laboratory-colonized, were assayed for susceptibilities to five insecticides (carbaryl, chlorpyrifos, malathion, permethrin, and pyrethrin) with an insecticide-treated, horizontally-oriented, Nylon 6,6 disk in a test tube. The pyrethrin was synergized using piperonyl butoxide (PBO). Flea mortality at two doses was recorded after 4 and 24 h exposures. The field strains from Texas and Florida tolerated carbaryl, chlorpyrifos, and malathion; and carbaryl and PBO-synergized pyrethrin, respectively. Tolerance was observed in a field strain from Kansas against malathion. Colonies from California and North Carolina were susceptible to malathion and PBO-synergized pyrethrin, and chlorpyrifos and permethrin, respectively, but three colonies showed tolerance. The insecticidal response of the California colony varied; when exposed to a chlorpyrifos dose of 10 mg (AI)/m2 for 24 h, at various times had mortality of 3-100%. With a PBO-synergized pyrethrin dose of 396 mg (AI)/ m2 for 4 h, the mortality ranged between 4.2 and 97%. Colonized strains were more susceptible than field strains at 4 h exposure to all insecticides except PBO-synergized pyrethrin. Colonized strains survived better in control tubes. The colony strains' susceptibility and variability are of considerable importance because these strains are used for flea product efficacy evaluations and bioassays. The differences in susceptibility between laboratory colonies and the field strains suggested development of both adaptation to colonization, and extensive, multiple cross-resistance to insecticides in field strains. Varying susceptibility of cat fleas may affect control success.