Insensitivity to the Spatial Repellent Action of Transfluthrin in Aedes aegypti: A Heritable Trait Associated with Decreased Insecticide Susceptibility

Background

New vector control paradigms expanding the use of spatial repellents are promising, but there are many gaps in our knowledge about how repellents work and how their long-term use might affect vector populations over time. Reported here are findings from a series of in vitro studies that investigated the plasticity and heritability of spatial repellent (SR) behaviors in Aedes aegypti exposed to airborne transfluthrin, including results that indicate a possible link between repellent insensitivity and insecticide resistance.

Methodology/principal findings

A dual-choice chamber system was used to observe directional flight behaviors in Aedes aegypti mosquitoes exposed to passively emanating transfluthrin vapors (1.35 mg/m³). Individual SR responder and SR non-responder mosquitoes were identified, collected and maintained separately according to their observed phenotype. Subsequent testing included re-evaluation of behavioral responses in some mosquito cohorts as well as testing the progeny of selectively bred responder and non-responder mosquito strains through nine generations. At baseline (F₀ generation), transfluthrin actively repelled mosquitoes in the assay system. F₀ mosquitoes repelled upon initial exposure to transfluthrin vapors were no more likely to be repelled again by subsequent exposure 24h later, but repelled mosquitoes allowed to rest for 48h were subsequently repelled at a higher proportion than was observed at baseline. Selective breeding of SR responders for nine generations did not change the proportion of mosquitoes repelled in any generation. However, selective breeding of SR non-responders did produce, after four generations, a strain of mosquitoes that was insensitive to the SR activity of transfluthrin. Compared to the SR responder strain, the SR insensitive strain also demonstrated decreased susceptibility to transfluthrin toxicity in CDC bottle bioassays and a higher frequency of the V1016I^kdr mutation.

Conclusions/significance

SR responses to volatile transfluthrin are complex behaviors with multiple determinants in Ae. aegypti. Results indicate a role for neurotoxic irritation of mosquitoes by sub-lethal doses of airborne chemical as a mechanism by which transfluthrin can produce SR behaviors in mosquitoes. Accordingly, how prolonged exposure to sub-lethal doses of volatile pyrethroids might impact insecticide resistance in natural vector populations, and how already resistant populations might respond to a given repellent in the field, are important considerations that warrant further monitoring and study. Results also highlight the critical need to develop new repellent active ingredients with novel mechanisms of action.     

Lethal and repellent effects of transfluthrin and metofluthrin used in portable blowers for personal protection against Ochlerotatus togoi and Aedes albopictus (Diptera: Culicidae)

The lethal efficacy of three vaporizing insecticides – 0.6% transfluthrin and 0.6% metofluthrin in portable battery‐powered blowers and 2.5% bioallethrin in electric mats – was assessed in unventilated and ventilated screened room conditions against Ochlerotatus togoi (Theobald) females. The mosquitoes were highly susceptible to transfluthrin and metofluthrin. Mean mortality rates of mosquitoes exposed to transfluthrin and metofluthrin in portable blowers ranged from 95 to 100% at a distance of 10 cm from mosquito cages in both room conditions; this was a >2.4‐fold greater mortality rate than for mosquitoes exposed to bioallethrin in electric mats under the same conditions (mean 40%). The three insecticides showed a decrease in mortality rate at a distance of 70 cm: rates were <5.0, 32.5, 85.0 and 90.0% for 10, 30, 60, and 120 min after exposure, respectively, for the same exposure periods in a closed room. In semi‐field repellent tests with human volunteers, 0.6% concentrations of transfluthrin and metofluthrin provided mean biting protection of 76.1 and 59.9% on legs, 68.5 and 52.1% on arms, 63.4 and 63.1% on the chest, and 87.1 and 52.9% on the face, respectively. In field tests, 0.6% transfluthrin effectively repelled Armigeres subalbatus and Aedes albopictus, with mean biting protection of 85.4 and 89.3% on exposed legs and arms, respectively, of the human volunteers. Metofluthrin at 0.6% also effectively repelled Ar. subalbatus, with mean biting protection levels of 71.8 and 73.5% on the legs and arms, respectively. There was no significant difference of repellent activity between the two vaporizing insecticides in the field.     

Efficacy of the botanical repellents geraniol,linalool, and citronella against mosquitoes

We determined the degree of personal protection provided by citronella, linalool, and geraniol in the form of commercially available candles or diffusers, both indoors and outdoors. Under the uniform conditions of the experiments, all substances repelled significantly more mosquitoes than the unprotected control. Furthermore, the repellents tested were more active when in the form of a continuous release diffuser than in candle form. All candles were 88g containing 5% of the active ingredient and all diffusers contained 20g of 100% active ingredient. Indoors, the repellency rate of citronella candles was only 14% while the repellency rate of citronella diffusers was 68%. The repellency of geraniol candles was 50% while the diffusers provided a repellency rate of 97%. No linalool candles were available for study but linalool diffusers repelled mosquitoes by 93%. Outdoors, citronella diffusers placed 6 m from mosquito traps repelled female mosquitoes by 22%, linalool repelled females by 58%, and geraniol repelled females by 75%. Trap catches were significantly reduced again when diffusers were placed 3 m from the traps. We concluded that geraniol had significantly more repellent activity than citronella or linalool in both indoor and outdoor settings.     

Understanding the Long-Lasting Attraction of Malaria Mosquitoes to Odor Baits

The use of odor baits for surveillance and control of malaria mosquitoes requires robust dispensing tools. In this study, the residual activity of a synthetic mosquito attractant blend dispensed from nylon or low density polyethylene (LDPE) sachets was evaluated at weekly intervals for one year without re-impregnation. The potential role of bacteria in modulating the attraction of mosquitoes to odor-treated nylon that had been used repeatedly over the one year study period, without re-impregnation, was also investigated. Significantly higher proportions of female Anopheles gambiae sensu stricto mosquitoes were consistently attracted to treated nylon strips than the other treatments, up to one year post-treatment. Additional volatile organic compounds and various bacterial populations were found on the treated nylon strips after one year of repeated use. The most abundant bacteria were Bacillus thuringiensis and Acinetobacter baumannii. Autoclaving of treated nylon strips prior to exposure had no effect on trap collections of laboratory-reared female An. Gambiae (P = 0.17) or wild female An. Gambiae sensu lato (P = 0.26) and Mansonia spp. (P = 0.17) mosquitoes. Trap catches of wild female An. Funestus (P < 0.001) and other anophelines (P < 0.007) were higher when treated strips had been autoclaved prior to deployment as opposed to when the treated nylon strips were not autoclaved. By contrast, wild female Culex mosquitoes were more strongly attracted to non-autoclaved compared to autoclaved treated nylon strips (P < 0.042). This study demonstrates the feasibility of using odor baits for sampling and surveillance of malaria as well as other mosquito vectors over prolonged periods of time. Preliminary evidence points towards the potential role of bacteria in sustaining prolonged use of nylon material for dispensing synthetic attractant odorants for host-seeking malaria and other mosquito vectors but further investigations are required.     

A comparative study on the efficacy of artesunate plus sulphadoxine/pyrimethamine versus artemether-lumefantrine in eastern Sudan

Background

The most important factor for effective zooprophylaxis in reducing malaria transmission is a predominant population of a strongly zoophilic mosquito, Anopheles arabiensis. The feeding preference behaviour of Anopheline mosquitoes was evaluated in odour-baited entry trap (OBET).

Methods

Mosquitoes were captured daily using odour-baited entry traps, light traps and hand catch both indoor and in pit traps. Experimental huts were used for release and recapture experiment. The mosquitoes collected were compared in species abundances.

Results

Anopheles arabiensis was found to account for over 99% of Anopheles species collected in the study area in Lower Moshi, Northern Tanzania. In experimental release/capture trials conducted at the Mabogini verandah huts, An. arabiensis was found to have higher exophilic tendency (80.7%) compared to Anopheles gambiae (59.7%) and Culex spp. (60.8%). OBET experiments conducted at Mabogini collected a total of 506 An. arabiensis in four different trials involving human, cattle, sheep, goat and pig. Odours from the cattle attracted 90.3% (243) compared to odours from human, which attracted 9.7% (26) with a significant difference at P = 0.005. Odours from sheep, goat and pig attracted 9.7%, 7.2% and 7.3%, respectively. Estimation of HBI in An. arabiensis collected from houses in three lower Moshi villages indicated lower ratios for mosquitoes collected from houses with cattle compared to those without cattles. HBI was also lower in mosquitoes collected outdoors (0.1–0.3) compared to indoor (0.4–0.9).

Conclusion

In discussing the results, reference has been made to observation of exophilic, zoophilic and feeding tendencies of An. arabiensis, which are conducive for zooprophylaxis. It is recommended that in areas with a predominant An. arabiensis population, cattle should be placed close to dwelling houses in order to maximize the effects of zooprophylaxis. Protective effects of human from malaria can further be enhanced by keeping cattle in surroundings of residences.     

Bait-fed adult Culex pipiens carry the larvicide Bacillus sphaericus to the larval habitat

Sugar meals of plant origin are a basic component of mosquito diet. We show that sugar baits have potential as vehicles for control agents. When Culex pipiens L. resting-sites were sprayed with sucrose solution, with or without incorporation of the larval toxicant Bacillus sphaericus Neide, 47% of female mosquitoes fed in situ. Dispersing B. sphaericus-carrier mosquitoes caused larval mortality in breeding-sites 60-100 m from the sprayed resting-sites. The effect was not seen where no adults rested in sparse vegetation above larval habitats. This approach may be useful for the application of biological control agents against mosquitoes in biotopes where the adults and larvae are juxtaposed.     

Evaluation of the pathogenicity and infectivity of entomopathogenic hypocrealean fungi,isolated from wild mosquitoes in Japan and Burkina Faso,against female adult Anopheles stephensi mosquitoes

We investigated the potential of entomopathogenic hypocrealean fungi that naturally occur in or on adult mosquitoes for use as biocontrol agents of vector mosquitoes. The fungi were isolated from wild mosquitoes collected in Japan and Burkina Faso using two isolation methods (with and without surface sterilization). Detected fungal species included Beauveria bassiana sensu lato, Isaria spp., Paecilomyces spp., Lecanicillium spp., and Simplicillium spp. These isolates were used in bioassays against adult female Anopheles stephensi mosquitoes. The median survival time ranged from 5.8 to 14.9 d (control, 17.0 d). Reduced survival times were observed in the isolates from surface-sterilized mosquitoes from Japan, with the isolate B. bassiana s.l. 60-2 exhibiting the highest virulence. This study indicates that adult mosquitoes are naturally infected with various entomopathogenic hypocrealean fungi, and that some of these isolates have the potential for use as fungal pesticides to control vector mosquitoes.     

Identification of super-infected Aedes triseriatus mosquitoes collected as eggs from the field and partial characterization of the infecting La Crosse viruses

Background

La Crosse virus (LACV) is a pathogenic arbovirus that is transovarially transmitted by Aedes triseriatus mosquitoes and overwinters in diapausing eggs. However, previous models predicted transovarial transmission (TOT) to be insufficient to maintain LACV in nature.

Results

To investigate this issue, we reared mosquitoes from field-collected eggs and assayed adults individually for LACV antigen, viral RNA by RT-PCR, and infectious virus. The mosquitoes had three distinct infection phenotypes: 1) super infected (SI+) mosquitoes contained infectious virus, large accumulations of viral antigen and RNA and comprised 17 of 17,825 (0.09%) of assayed mosquitoes, 2) infected mosquitoes (I+) contained no detectable infectious virus, lesser amounts of viral antigen and RNA, and comprised 3.7% of mosquitoes, and 3) non-infected mosquitoes (I-) contained no detectable viral antigen, RNA, or infectious virus and comprised 96.21% of mosquitoes. SI+ mosquitoes were recovered in consecutive years at one field site, suggesting that lineages of TOT stably-infected and geographically isolated Ae. triseriatus exist in nature. Analyses of LACV genomes showed that SI+ isolates are not monophyletic nor phylogenetically distinct and that synonymous substitution rates exceed replacement rates in all genes and isolates. Analysis of singleton versus shared mutations (Fu and Li's F*) revealed that the SI+ LACV M segment, with a large and significant excess of intermediate-frequency alleles, evolves through disruptive selection that maintains SI+ alleles at higher frequencies than the average mutation rate. A QTN in the LACV NSm gene was detected in SI+ mosquitoes, but not in I+ mosquitoes. Four amino acid changes were detected in the LACV NSm gene from SI+ but not I+ mosquitoes from one site, and may condition vector super infection. In contrast to NSm, the NSs sequences of LACV from SI+ and I+ mosquitoes were identical.

Conclusions

SI+ mosquitoes may represent stabilized infections of Ae. triseriatus mosquitoes, which could maintain LACV in nature. A gene-for-gene interaction involving the viral NSm gene and a vector innate immune response gene may condition stabilized infection.     

Deletion of the NSm Virulence Gene of Rift Valley Fever Virus Inhibits Virus Replication in and Dissemination from the Midgut of Aedes aegypti Mosquitoes

Background

Previously, we investigated the role of the Rift Valley fever virus (RVFV) virulence genes NSs and NSm in mosquitoes and demonstrated that deletion of NSm significantly reduced the infection, dissemination, and transmission rates of RVFV in Aedes aegypti mosquitoes. The specific aim of this study was to further characterize midgut infection and escape barriers of RVFV in Ae. aegypti infected with reverse genetics-generated wild type RVFV (rRVF-wt) or RVFV lacking the NSm virulence gene (rRVF-ΔNSm) by examining sagittal sections of infected mosquitoes for viral antigen at various time points post-infection.

Methodology and Principal Findings

Ae. aegypti mosquitoes were fed an infectious blood meal containing either rRVF-wt or rRVF-ΔNSm. On days 0, 1, 2, 3, 4, 6, 8, 10, 12, and 14 post-infection, mosquitoes from each experimental group were fixed in 4% paraformaldehyde, paraffin-embedded, sectioned, and examined for RVFV antigen by immunofluorescence assay. Remaining mosquitoes at day 14 were assayed for infection, dissemination, and transmission. Disseminated infections were observed in mosquitoes as early as three days post infection for both virus strains. However, infection rates for rRVF-ΔNSm were statistically significantly less than for rRVF-wt. Posterior midgut infections in mosquitoes infected with rRVF-wt were extensive, whereas midgut infections of mosquitoes infected with rRVF-ΔNSm were confined to one or a few small foci.

Conclusions/Significance

Deletion of NSm resulted in the reduced ability of RVFV to enter, replicate, and disseminate from the midgut epithelial cells. NSm appears to have a functional role in the vector competence of mosquitoes for RVFV at the level of the midgut barrier.     

Laboratory studies on the olfactory behaviour of Anopheles quadriannulatus

The host preference of Anopheles quadriannulatus Theobald (Diptera: Culicidae), the zoophilic member of the malaria mosquito complex Anopheles gambiae Giles, was investigated in a dual‐choice olfactometer. Naïve female mosquitoes were exposed to CO₂, acetone, 1‐octen‐3‐ol, and skin emanations from cows and humans in various combinations. Their behavioural responses were recorded when they had entered one of either upwind traps from where the odours were being released. The mosquitoes did not respond to CO₂ when released at human or cattle equivalent concentrations. Too few mosquitoes responded to acetone to allow for a statistical analysis. The combination of CO₂+ 1‐octen‐3‐ol was repellent. Cow odour alone was slightly attractive, but there was a synergistic attractive effect of cow odour + CO₂. Surprisingly, the mosquitoes were attracted to human odour, and in a choice situation human odour was selected above cow odour + CO₂. Anthropophilic An. gambiae Giles s.s. was repelled by cow odour + CO₂ in contrast to An. quadriannulatus. In a choice situation, both mosquito species selected human odour above cow odour + CO₂. The implications of these results are discussed in the light of recent behavioural data from the field.     

Olfactory memory in the mosquito Culex quinquefasciatus

The cosmotropical urban mosquito Culex quinquefasciatus Say (Diptera: Culicidae) uses chemical cues to locate suitable water pools for oviposition. Although gravid females are innately attracted to or repelled by certain compounds, this study found that an individual mosquito's preferences for these odours could be altered greatly by prior experience. Mosquitoes reared in water containing skatole, at a level normally repellent to ovipositing females, preferred to oviposit in water containing that compound rather than in water with an otherwise attractive odour compound (P-cresol). This behaviour occurred regardless of whether mosquitoes were tested individually or in groups of up to 50 per cage. The F1 progeny of conditioned mosquitoes did not exhibit the parental preference, but were as susceptible to conditioning as their parents. Moreover, rearing mosquitoes in infusions of hay or animal (guinea-pig) faeces produced a similar although less dramatic change, such that the innate propensity for hay infusion could be cancelled by rearing in guinea-pig faeces infusion. The results demonstrated a change in odour preference by Cx. quinquefasciatus following exposure to the odour during development or pupal eclosion, suggesting that some form of larval conditioning or early adult imprinting occurred. Precisely when that conditioning occurred remains to be determined.     

Evaluation of kill strips on boll weevil (Coleoptera: Curculionidae) mortality in pheromone traps and impact on weevil escape

A field study examined the temporal patterns of boll weevil mortality provided by two commercially available kill strips, Hercon VaporTape II and Plato Insecticide Strip, and to evaluate the impacts of these devices on weevil escape from traps. Both types of kill strips produced similar levels of weevil mortality with the exception of the last two inspection intervals (30 and 46 h after continual exposure to kill strips). At these intervals, the Plato Strip produced significantly higher mortality than the Hercon strip; however, these differences were numerically small (10 and 6%, respectively). Both types of kill strips produced a high level of weevil mortality in traps (> 90%) after 46 h of exposure. On average, 5-8% of weevils escaped from traps whether a kill strip was present or absent. A strong temporal pattern of escape was observed, with > or = 90% of escape occurring within the first hour after weevils were introduced into traps. Because > or = 90% of escape occurred within the first hour weevils were in the traps and < 3% of weevils died during the first hour of exposure to kill strips in traps, use of kill strips in large-scale boll weevil management programs is not justified on the basis of reduced weevil escape.     

Maiden outbreak of chikungunya in dongguan city, guangdong province, china: epidemiological characteristics

Background

This study was conducted to identify epidemiological characteristics of the first documented CHIK fever outbreak in China and evaluate the effect of the preventive measures taken.

Methodology/Principal Findings

From September 1 to October 29, 2010, China''s first documented outbreak of CHIK fever occurred in the Xincun community of Wanjiang District of Dongguan city, Guangdong province; 253 case-patients were recorded, of which 129 were laboratory confirmed, with an attack rate of 1%. Before September 18^th the number of CHIK fever cases remained relatively low in the Xincun community; from September 19^th onwards, the number of cases increased drastically, with an outbreak peak on October 4^th. Cases were distributed across nine small village groups in the Xincun community, with an attack rate of 0–12% at the village level. The household attack rates ranged between 20% and 100%. No significant difference was found in the attack rate between males and females. There was a significant difference in the attack rate in different age groups (chi-square = 18.35, p = 0.005); highest in patients aged 60 years or older and the lowest in patients aged under 10. The major clinical characteristics of patients are fever (100%), joint pain (79%) and rash (54%). Phylogenetic analysis of the E1 gene on the five earliest confirmed cases showed that the strains of CHIKV isolated from their sera were highly homologous (up to 99%) with isogeneic strains isolated in Thailand in 2009. After control measures were taken, including killing adult mosquitoes and cleaning breeding habitats of Aedes mosquitoes, the Breteau index and Mosq-ovitrap index decreased rapidly, and the outbreak ended on October 29.

Conclusion/Significance

The infection source of the outbreak was imported. Cases showed obvious temporal, spatial, and population aggregation during the outbreak. Comprehensive control measures based on reducing the density of Aedes mosquitoes were effective in controlling the epidemic.     

Fine scale spatiotemporal clustering of dengue virus transmission in children and Aedes aegypti in rural Thai villages

Background

Based on spatiotemporal clustering of human dengue virus (DENV) infections, transmission is thought to occur at fine spatiotemporal scales by horizontal transfer of virus between humans and mosquito vectors. To define the dimensions of local transmission and quantify the factors that support it, we examined relationships between infected humans and Aedes aegypti in Thai villages.

Methodology/Principal Findings

Geographic cluster investigations of 100-meter radius were conducted around DENV-positive and DENV-negative febrile “index” cases (positive and negative clusters, respectively) from a longitudinal cohort study in rural Thailand. Child contacts and Ae. aegypti from cluster houses were assessed for DENV infection. Spatiotemporal, demographic, and entomological parameters were evaluated. In positive clusters, the DENV infection rate among child contacts was 35.3% in index houses, 29.9% in houses within 20 meters, and decreased with distance from the index house to 6.2% in houses 80–100 meters away (p<0.001). Significantly more Ae. aegypti were DENV-infectious (i.e., DENV-positive in head/thorax) in positive clusters (23/1755; 1.3%) than negative clusters (1/1548; 0.1%). In positive clusters, 8.2% of mosquitoes were DENV-infectious in index houses, 4.2% in other houses with DENV-infected children, and 0.4% in houses without infected children (p<0.001). The DENV infection rate in contacts was 47.4% in houses with infectious mosquitoes, 28.7% in other houses in the same cluster, and 10.8% in positive clusters without infectious mosquitoes (p<0.001). Ae. aegypti pupae and adult females were more numerous only in houses containing infectious mosquitoes.

Conclusions/Significance

Human and mosquito infections are positively associated at the level of individual houses and neighboring residences. Certain houses with high transmission risk contribute disproportionately to DENV spread to neighboring houses. Small groups of houses with elevated transmission risk are consistent with over-dispersion of transmission (i.e., at a given point in time, people/mosquitoes from a small portion of houses are responsible for the majority of transmission).     

Toward development of neem-based repellents against the Scottish Highland biting midge Culicoides impunctatus

Oil of neem, from the tree Azadirachta indica A. Juss (Meliaceae), was evaluated for repellent and antifeedant activity against Culicoides biting midges (Diptera: Ceratopogonidae), by three complementary methods with serial dilutions. Electroantennograms revealed the sensitivity of Culicoides nubeculosus (Meigen) females to neem > or = 0.10%. Culicoides impunctatus Goetghebuer females were repelled by > or = 1% in a Y-tube olfactometer, Using a membrane feeder for wild-caught parous females of C. impunctatus, the proportion blood-feeding was significantly reduced by topical applications of neem oil > or = 0.10% concentrations, with blood-feeding completely prevented by > or =1%. On the basis of these response data, we developed 2% neem-based formulations for personal protection against biting midges.     

Attractiveness of synthetic volatile blends of flowering rice panicles to Trigonotylus caelestialium (Kirkaldy) (Heteroptera: Miridae)

The attractiveness of 22 synthetic volatile blends or seven individual chemicals emitted from flowering rice panicles to a rice leaf bug, Trigonotylus caelestialium (Kirkaldy), were investigated with an olfactometer to identify the active compounds responsible for the invasion of the bugs into paddy fields. n‐Decanal attracted only male bugs, whereas β‐caryophyllene attracted only females. β‐Elemene repelled males and methyl benzoate marginally repelled females. The other chemicals did not attract or repel male and female bugs at all. Two‐, 3‐, 4‐ and 5‐component blends of β‐caryophyllene, n‐decanal, n‐tridecene, methyl salicylate and geranyl acetone were attractive to neither females nor males. Two‐component blends comprised of β‐caryophyllene and methyl salicylate, or n‐decanal and methyl salicylate, marginally repelled females. The three‐component blend comprised of β‐caryophyllene, n‐decanal and geranyl acetone marginally repelled females. The five‐component blend comprised of β‐caryophyllene, n‐decanal, n‐tridecene, methyl salicylate and geranyl acetone repelled males. The seven‐component blend comprised of β‐caryophyllene, n‐decanal, n‐tridecene, methyl salicylate, geranyl acetone, methyl benzoate and β‐elemene attracted female bugs and marginally attracted male bugs. Six‐component blends without any one of these seven components were not attractive to the bugs although the six‐component blend without n‐decanal was marginally attractive to females. The six‐component blend without n‐tridecene repelled males. These findings suggest that mixtures of these seven compounds play an important role in the attractiveness of flowering rice panicles to both sexes of the bugs, although the attractiveness of individual compounds differs between sexes.     

Laboratory testing of a lethal ovitrap for Aedes aegypti

Laboratory tests were conducted to determine the feasibility of making the mosquito ovitrap lethal to Aedes aegypti (L.) when they attempt to oviposit in the trap. Heavy-weight velour paper strips (2.54 x 11 cm) were used as an alternative to the wooden paddle normally provided as a substrate for mosquito oviposition. The paper strips were pretreated with insecticide solutions and allowed to dry before being used in oviposition cups of 473 ml capacity, filled with water initially to within 2.5 cm of the brim. Insecticides chosen for their quick knock-down efficacy were bendiocarb 76% WP (1.06 mg a.i./strip) and four pyrethroids: permethrin 25% WP (0.16 mg a.i./strip), deltamethin 4.75% SC (0.87 mg a.i./strip), cypermethrin 40% WP (2.81 mg a.i./strip), and cyfluthrin 20% WP (0.57 mg a.i./ strip). For experimental evaluation, two oviposition cups (one with an insecticide-treated strip and one with an untreated strip) were placed in cages (cubic 30 cm) with gravid female Ae. aegypti mosquitoes (aged 6-8 days) from a susceptible laboratory strain. Mortality-rates of female mosquitoes were 45% for bendiocarb, 47% for permethrin, 98% for deltamethrin, 100% for cypermethrin, and 100% for cyfluthrin. Young instar larvae added to the treated cups died within 2h. After water evaporation from the cups for 38 days, fresh mosquito females had access to previously submerged portions of the velour paper paddle, and mortality rates of 59% or more occurred. Cups that had water (360 ml) dripped into them, to simulate rain, produced female mosquito mortality rates of > 50% and all larvae died within 3 h of being added. These tests demonstrate that the ovitrap can be made lethal to both adults and larvae by insecticidal treatment of the ovistrip. Field efficacy trials are underway in Brazil to access the impact of this simple, low-cost, environmentally benign approach on populations of the dengue vector Ae. aegypti.     

Uterine motor alterations and estrous cycle disturbances associated with colonic inflammation in the rat

The impact of colitis on uterine contractility and estrous cycle was investigated after intracolonic administration of 2,4,6-trinitrobenzenesulfonic acid (TNBS) in rats. Colitis severity was assessed by macroscopic damage scoring (MDS) 4 days after TNBS, and myeloperoxidase (MPO) activity was measured in both colon and uterus of control and colitic rats. Estrous cycle stages were determined by vaginal smears and histology, and uterine contractility was assessed in vitro on longitudinal and circular strips. In control rats, uterine MPO activity varied markedly during the cycle and peaked around estrus. In rats with moderate colitis [MDS < 5, 3.1 +/- 0.2 (mean +/- SE)], uterine MPO decreased by 61% compared with estrus control, without disruption of the cycle. Frequency of spontaneous contractions was reduced by 32% in circular muscle. Contractile responses to KCl and carbachol were not affected, whereas maximal response to oxytocin decreased by 47% in the longitudinal muscle. In rats with severe colitis (MDS > 5, 6.0 +/- 0.2), uterine MPO was reduced by 96% and estrous cycle was disrupted. Spontaneous contractility was impaired in circular strips, and a 39% decrease in the contraction frequency occurred in the longitudinal strips. Circular strips did not contract to KCl or carbachol; however, longitudinal strips had maximal responses to KCl, carbachol, and oxytocin reduced by 36%, 27%, and 46%, respectively. Estrogen replacement protected the uterine responses to carbachol in colitic rats, whereas oxytocin responses remained depressed. These data indicate that colonic inflammation can influence both spontaneous and evoked uterine contractility, in relation to estrous cycle disturbances, impaired estradiol production, and functional alterations of myometrial cells.     

Insecticide susceptibility status in Mexican populations of Stegomyia aegypti (= Aedes aegypti): a nationwide assessment

In Mexico, mosquito vector‐borne diseases are of public health concern as a result of their impact on human morbidity and mortality. The use of insecticides against adult mosquitoes is one of the most common ways of controlling mosquito population densities. However, the use of these compounds has resulted in the development of insecticide resistance. The aim of this study was to estimate susceptibility to six pyrethroids, two carbamates and two organophosphates in Mexican populations of Stegomyia aegypti (Linnaeus, 1762) (= Aedes aegypti) (Diptera: Culicidae) mosquitoes. Bottle insecticide susceptibility tests, with 1 h exposure, were performed on adult mosquitoes from 75 localities across 28 states. At 30 min of exposure, the proportion of fallen mosquitoes was recorded. After 60 min of exposure, mosquitoes were recovered in non‐treated containers and mortality was determined at 24 h after the set‐up of the experiment. In general, the carbamate insecticides represented the most effective group in terms of the proportion of mosquitoes fallen at 30 min (72–100%) and 24‐h mortality (97–100%). High and widespread resistance to pyrethroids Types I and II and, to a lesser extent, to organophosphates was observed. Insecticide susceptibility among and within states was highly variable.