The TIRS trial: Enrollment procedures and baseline characterization of a pediatric cohort to quantify the epidemiologic impact of targeted indoor residual spraying on Aedes-borne viruses in Merida, Mexico.

Aedes mosquito-borne viruses (ABVs) place a substantial strain on public health resources in the Americas. Vector control of Aedes mosquitoes is an important public health strategy to decrease or prevent spread of ABVs. The ongoing Targeted Indoor Residual Spraying (TIRS) trial is an NIH-sponsored clinical trial to study the efficacy of a novel, proactive vector control technique to prevent dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV) infections in the endemic city of Merida, Yucatan, Mexico. The primary outcome of the trial is laboratory-confirmed ABV infections in neighborhood clusters. Despite the difficulties caused by the COVID-19 pandemic, by early 2021 the TIRS trial completed enrollment of 4,792 children aged 2-15 years in 50 neighborhood clusters which were allocated to control or intervention arms via a covariate-constrained randomization algorithm. Here, we describe the makeup and ABV seroprevalence of participants and mosquito population characteristics in both arms before TIRS administration. Baseline surveys showed similar distribution of age, sex, and socio-economic factors between the arms. Serum samples from 1,399 children were tested by commercially available ELISAs for presence of anti-ABV antibodies. We found that 45.1% of children were seropositive for one or more flaviviruses and 24.0% were seropositive for CHIKV. Of the flavivirus-positive participants, most were positive for ZIKV-neutralizing antibodies by focus reduction neutralization testing which indicated a higher proportion of participants with previous ZIKV than DENV infections within the cohort. Both study arms had statistically similar seroprevalence for all viruses tested, similar socio-demographic compositions, similar levels of Ae. aegypti infestation, and similar observed mosquito susceptibility to insecticides. These findings describe a population with a high rate of previous exposure to ZIKV and lower titers of neutralizing antibodies against DENV serotypes, suggesting susceptibility to future outbreaks of flaviviruses is possible, but proactive vector control may mitigate these risks.

Residual efficacy of the neonicotinoid insecticide clothianidin against pyrethroid-resistant Aedes aegypti.

BACKGROUND: Here we report the residual efficacy of the neonicotinoid insecticide clothianidin against pyrethroid-resistant Aedes aegypti. We first conducted a range-finding evaluation of clothianidin on three different substrates (wall, wood, cloth) using three doses (100, 300 and 600 mg a.i. m-2 ) and conducting World Health Organization (WHO) cone bioassays to assess acute (24 h) and delayed (up to 7 days) mortality. In experimental houses located in Merida (Mexico) and using free-flying pyrethroid-resistant Ae. aegypti females, we quantified the acute and delayed mortality after a 24-h exposure to the targeted indoor residual spraying (TIRS) of two clothianidin doses (100 and 300 mg a.i. m-2 ). RESULTS: Range-finding studies with WHO cones showed low (<50%) acute mortality for all surfaces, doses and times post spraying. Delayed mortality was higher, with average values above or close to the 60% mark (and 95% confidence interval estimates crossing 80% for the 600 mg a.i. m-2 dose). In experimental houses, a similar low acute mortality was quantified (range of mortality across 12 months was 2-44% for 100 mg a.i. m-2 and 8-61% for 300 mg a.i/m2 ). However, delayed mortality showed a strong effect of clothianidin on free-flying Ae. aegypti, with values above 80% up to 7 months post-TIRS. CONCLUSION: Novel residual insecticide molecules have a promising outlook for Ae. aegypti control and can contribute to the expansion and adoption of TIRS in urban areas. clothianidin can contribute to the control of resistant Ae. aegypti and provide residual control for up to 7 months after application. © 2022 Society of Chemical Industry.

Preventive residual insecticide applications successfully controlled Aedes aegypti in Yucatan, Mexico.

Insecticide-based approaches remain a key pillar for Aedes-borne virus (ABV, dengue, chikungunya, Zika) control, yet they are challenged by the limited effect of traditional outdoor insecticide campaigns responding to reported arboviral cases and by the emergence of insecticide resistance in mosquitoes. A three-arm Phase II unblinded entomological cluster randomized trial was conducted in Merida, Yucatan State, Mexico, to quantify the entomological impact of targeted indoor residual spraying (TIRS, application of residual insecticides in Ae. aegypti indoor resting sites) applied preventively 2 months before the beginning of the arbovirus transmission season. Trial arms involved the use of two insecticides with unrelated modes of action (Actellic 300CS, pirimiphos-methyl, and SumiShield 50WG, clothianidin) and a control arm where TIRS was not applied. Entomological impact was quantified by Prokopack adult collections performed indoors during 10 min per house. Regardless of the insecticide, conducting a preventive TIRS application led to significant reductions in indoor Ae. aegypti densities, which were maintained at the same levels as in the low arbovirus transmission period (Actellic 300CS reduced Ae. aegypti density up to 8 months, whereas SumiShield 50WG up to 6 months). The proportional reduction in Ae. aegypti abundance in treatment houses compared to control houses was 50-70% for Actellic 300CS and 43-63% for SumiShield 50WG. Total operational costs including insecticide ranged from US$4.2 to US$10.5 per house, depending on the insecticide cost. Conducting preventive residual insecticide applications can maintain Ae. aegypti densities at low levels year-round with important implications for preventing ABVs in the Americas and beyond.

Natural Aedes-Borne Virus Infection Detected in Male Adult Aedes aegypti (Diptera: Culicidae) Collected From Urban Settings in Mérida, Yucatán, México.

Aedes-borne viruses (ABVs) such as dengue (DENV), chikungunya (CHIKV), and Zika (ZIKV) contribute significantly to the global burden of infectious diseases, disproportionately affecting disadvantaged populations from tropical and subtropical urban areas. ABVs can be transmitted from female mosquitoes to their progeny by vertical transmission via transovarial and/or trans-egg vertical transmission and contribute to the maintenance of infected-mosquito populations year-round in endemic regions. This study describes the natural infection rate of DENV, CHIKV, and ZIKV in field-caught male Aedes (Sergentomyia) aegypti (Linnaeus) mosquitoes from Mérida, Yucatán, México, as a proxy for the occurrence of vertical virus transmission. We used indoor sequential sampling with Prokopack aspirators to collect all mosquitoes inside houses from ABV hotspots areas. Collections were performed in a DENV and CHIKV post-epidemic phase and during a period of active ZIKV transmission. We individually RT-qPCR tested all indoor collected Ae. aegypti males (1,278) followed by Sanger sequencing analysis for final confirmation. A total of 6.7% male mosquitoes were positive for ABV (CHIKV = 5.7%; DENV = 0.9%; ZIKV = 0.1%) and came from 21.0% (30/143) houses infested with males. Most ABV-positive male mosquitoes were positive for CHIKV (84.8%). The distribution of ABV-positive Ae. aegypti males was aggregated in a few households, with two houses having 11 ABV-positive males each. We found a positive association between ABV-positive males and females per house. These findings suggested the occurrence of vertical arbovirus transmission within the mosquito populations in an ABV-endemic area and, a mechanism contributing to viral maintenance and virus re-emergence among humans in post-epidemic periods.

Pilot trial using mass field-releases of sterile males produced with the incompatible and sterile insect techniques as part of integrated Aedes aegypti control in Mexico.

BACKGROUND: The combination of Wolbachia-based incompatible insect technique (IIT) and radiation-based sterile insect technique (SIT) can be used for population suppression of Aedes aegypti. Our main objective was to evaluate whether open-field mass-releases of wAlbB-infected Ae. aegypti males, as part of an Integrated Vector Management (IVM) plan led by the Mexican Ministry of Health, could suppress natural populations of Ae. aegypti in urbanized settings in south Mexico. METHODOLOGY/PRINCIPAL FINDINGS: We implemented a controlled before-and-after quasi-experimental study in two suburban localities of Yucatan (Mexico): San Pedro Chimay (SPC), which received IIT-SIT, and San Antonio Tahdzibichén used as control. Release of wAlbB Ae. aegypti males at SPC extended for 6 months (July-December 2019), covering the period of higher Ae. aegypti abundance. Entomological indicators included egg hatching rates and outdoor/indoor adult females collected at the release and control sites. Approximately 1,270,000 lab-produced wAlbB-infected Ae. aegypti males were released in the 50-ha treatment area (2,000 wAlbB Ae. aegypti males per hectare twice a week in two different release days, totaling 200,000 male mosquitoes per week). The efficacy of IIT-SIT in suppressing indoor female Ae. aegypti density (quantified from a generalized linear mixed model showing a statistically significant reduction in treatment versus control areas) was 90.9% a month after initiation of the suppression phase, 47.7% two months after (when number of released males was reduced in 50% to match local abundance), 61.4% four months after (when initial number of released males was re-established), 88.4% five months after and 89.4% at six months after the initiation of the suppression phase. A proportional, but lower, reduction in outdoor female Ae. aegypti was also quantified (range, 50.0-75.2% suppression). CONCLUSIONS/SIGNIFICANCE: Our study, the first open-field pilot implementation of Wolbachia IIT-SIT in Mexico and Latin-America, confirms that inundative male releases can significantly reduce natural populations of Ae. aegypti. More importantly, we present successful pilot results of the integration of Wolbachia IIT-SIT within a IVM plan implemented by Ministry of Health personnel.

Mosquito species (Diptera: culicidae) collected after tropical storm cristobal in Merida, Yucatan, South-east Mexico.

After the tropical storm Cristobal, we performed special adult entomological collections in the peri-domicile of 35 houses from 25 neighborhoods of Mérida, Yucatan, Mexico in response to complaints from the community about an increased nuisance due to an abundance of mosquitoes. A total of 1,275 specimens from four genera and 13 species were collected: Aedes taeniorhynchus (92%), Culex quinquefasciatus (72%), Aedes aegypti (72%), Psorophora mexicana (36%), Psorophora cyanescens (32%), Aedes scapularis (24%), Culex nigripalpus (24%), Aedes albopictus (8%), Psorophora ferox (4%), Haemagogus equinus (4%), Aedes trivittatus (4%), Culex coronator (4%), Culex iolambdis (4%). From these collections, the increased mosquito nuisance was mainly the result of invasive species such as Aedes taeniorhynchus and Psorophora. City wide, vehicle mounted ULV spraying was performed by the MoH and the municipality of Merida to control adult mosquito populations. We report Culex iolambdis for the first time in Merida and Psorophora mexicana for the state of Yucatan.

Efficacy of targeted indoor residual spraying with the pyrrole insecticide chlorfenapyr against pyrethroid-resistant Aedes aegypti.

BACKGROUND: There is an increased need to mitigate the emergence of insecticide resistance and incorporate new formulations and modes of application to control the urban vector Aedes aegypti. Most research and development of insecticide formulations for the control of Ae. aegypti has focused on their peridomestic use as truck-mounted ULV-sprays or thermal fogs despite the widespread knowledge that most resting Ae. aegypti are found indoors. A recent modification of indoor residual spraying (IRS), termed targeted IRS (TIRS) works by restricting applications to 1.5 m down to the floor and on key Ae. aegypti resting sites (under furniture). TIRS also opens the possibility of evaluating novel residual insecticide formulations currently being developed for malaria IRS. METHODS: We evaluated the residual efficacy of chlorfenapyr, formulated as Sylando 240SC, for 12 months on free-flying field-derived pyrethroid-resistant Ae. aegypti using a novel experimental house design in Merida, Mexico. On a monthly basis, 600 female Ae. aegypti were released into the houses and left indoors with access to sugar solution for 24 hours. After the exposure period, dead and alive mosquitoes were counted in houses treated with chlorfenapyr as well as untreated control houses to calculate 24-h mortality. An evaluation for these exposed cohorts of surviving mosquitoes was extended up to seven days under laboratory conditions to quantify "delayed mortality". RESULTS: Mean acute (24-h) mortality of pyrethroid-resistant Ae. aegypti ranged 80-97% over 5 months, dropping below 30% after 7 months post-TIRS. If delayed mortality was considered (quantifying mosquito mortality up to 7 days after exposure), residual efficacy was above 90% for up to 7 months post-TIRS application. Generalized Additive Mixed Models quantified a residual efficacy of chlorfenapyr of 225 days (ca. 7.5 months). CONCLUSIONS: Chlorfenapyr represents a new option for TIRS control of Ae. aegypti in urban areas, providing a highly-effective time of protection against indoor Ae. aegypti females of up to 7 months.

Protective effect of house screening against indoor Aedes aegypti in Mérida, Mexico: A cluster randomised controlled trial.

OBJECTIVE: To evaluate the protective effect of house screening (HS) on indoor Aedes aegypti infestation, abundance and arboviral infection in Merida, Mexico. METHODS: In 2019, we performed a cluster randomised controlled trial (6 control and 6 intervention areas: 100 households/area). Intervention clusters received permanently fixed fiberglass HS on all windows and doors. The study included two cross-sectional entomologic surveys, one baseline (dry season in May 2019) and one post-intervention (PI, rainy season between September and October 2019). The presence and number of indoor Aedes females and blood-fed females (indoor mosquito infestation) as well as arboviral infections with dengue (DENV) and Zika (ZIKV) viruses were evaluated in a subsample of 30 houses within each cluster. RESULTS: HS houses had significantly lower risk for having Aedes aegypti female mosquitoes (odds ratio [OR] = 0.56, 95% CI 0.33-0.97, p = 0.04) and blood-fed females (OR = 0.53, 95% CI 0.28-0.97, p = 0.04) than unscreened households from the control arm. Compared to control houses, HS houses had significantly lower indoor Ae. aegypti abundance (rate ratio [RR] = 0.50, 95% CI 0.30-0.83, p = 0.01), blood-fed Ae. aegypti females (RR = 0.48, 95% CI 0.27-0.85, p = 0.01) and female Ae. aegypti positive for arboviruses (OR = 0.29, 95% CI 0.10-0.86, p = 0.02). The estimated intervention efficacy in reducing Ae. aegypti arbovirus infection was 71%. CONCLUSIONS: These results provide evidence supporting the use of HS as an effective pesticide-free method to control house infestations with Aedes aegypti and reduce the transmission of Aedes-transmitted viruses such as DENV, chikungunya (CHIKV) and ZIKV.

The entomological impact of passive metofluthrin emanators against indoor Aedes aegypti: A randomized field trial.

BACKGROUND: In the absence of vaccines or drugs, insecticides are the mainstay of Aedes-borne disease control. Their utility is challenged by the slow deployment of resources, poor community compliance and inadequate household coverage. Novel application methods are required. METHODOLOGY AND PRINCIPAL FINDINGS: A 10% w/w metofluthrin "emanator" that passively disseminates insecticide from an impregnated net was evaluated in a randomized trial of 200 houses in Mexico. The devices were introduced at a rate of 1 per room and replaced at 3-week intervals. During each of 7 consecutive deployment cycles, indoor resting mosquitoes were sampled using aspirator collections. Assessments of mosquito landing behaviours were made in a subset of houses. Pre-treatment, there were no differences in Aedes aegypti indices between houses recruited to the control and treatment arms. Immediately after metofluthrin deployment, the entomological indices between the trial arms diverged. Averaged across the trial, there were significant reductions in Abundance Rate Ratios for total Ae. aegypti, female abundance and females that contained blood meals (2.5, 2.4 and 2.3-times fewer mosquitoes respectively; P<0.001). Average efficacy was 60.2% for total adults, 58.3% for females, and 57.2% for blood-fed females. The emanators also reduced mosquito landings by 90% from 12.5 to 1.2 per 10-minute sampling period (P<0.05). Homozygous forms of the pyrethroid resistant kdr alleles V410L, V1016L and F1534C were common in the target mosquito population; found in 39%, 24% and 95% of mosquitoes collected during the trial. CONCLUSIONS/SIGNIFICANCE: This is the first randomized control trial to evaluate the entomological impact of any volatile pyrethroid on urban Ae. aegypti. It demonstrates that volatile pyrethroids can have a sustained impact on Ae. aegypti population densities and human-vector contact indoors. These effects occur despite the presence of pyrethroid-resistant alleles in the target population. Formulations like these may have considerable utility for public health vector control responses.

Insecticide-treated house screening protects against Zika-infected Aedes aegypti in Merida, Mexico.

BACKGROUND: The integration of house-screening and long-lasting insecticidal nets, known as insecticide-treated screening (ITS), can provide simple, safe, and low-tech Aedes aegypti control. Cluster randomised controlled trials in two endemic localities for Ae. aegypti of south Mexico, showed that ITS conferred both, immediate and sustained (~2 yr) impact on indoor-female Ae. aegypti infestations. Such encouraging results require further validation with studies quantifying more epidemiologically-related endpoints, including arbovirus infection in Ae. aegypti. We evaluated the efficacy of protecting houses with ITS on Ae. aegypti infestation and arbovirus infection during a Zika outbreak in Merida, Yucatan, Mexico. METHODOLOGY/PRINCIPAL FINDINGS: A two-arm cluster-randomised controlled trial evaluated the entomological efficacy of ITS compared to the absence of ITS (with both arms able to receive routine arbovirus vector control) in the neighbourhood Juan Pablo II of Merida. Cross-sectional entomological surveys quantified indoor adult mosquito infestation and arbovirus infection at baseline (pre-ITS installation) and throughout two post-intervention (PI) surveys spaced at 6-month intervals corresponding to dry/rainy seasons over one year (2016-2017). Household-surveys assessed the social reception of the intervention. Houses with ITS were 79-85% less infested with Aedes females than control houses up to one-year PI. A similar significant trend was observed for blood-fed Ae. aegypti females (76-82%). Houses with ITS had significantly less infected female Ae. aegypti than controls during the peak of the epidemic (OR = 0.15, 95%CI: 0.08-0.29), an effect that was significant up to a year PI (OR = 0.24, 0.15-0.39). Communities strongly accepted the intervention, due to its perceived mode of action, the prevalent risk for Aedes-borne diseases in the area, and the positive feedback from neighbours receiving ITS. CONCLUSIONS/SIGNIFICANCE: We show evidence of the protective efficacy of ITS against an arboviral disease of major relevance, and discuss the relevance of our findings for intervention adoption.

Evaluating Over-the-Counter Household Insecticide Aerosols for Rapid Vector Control of Pyrethroid-Resistant Aedes aegypti.

Vector control methods that mobilize and impact rapidly during dengue, Zika, and chikungunya outbreaks are urgently needed in urban contexts. We investigated whether one person using a handheld aerosolized insecticide could achieve efficacy levels comparable to targeted indoor residual spraying (TIRS), using pyrethroid-resistant Aedes aegypti in a semi-field setting with experimental houses in Mexico. The insecticide product (H24, a carbamate and pyrethroid mixture), available over-the-counter locally, was sprayed only on known Ae. aegypti-resting surfaces, for example, walls less than 1.5 m and dark hidden areas. In six identical houses with paired bedrooms, one bedroom was treated, and the other remained an untreated control. Each week for 8 weeks, 100 female pyrethroid-resistant Ae. aegypti were released in each bedroom and followed up daily. Mortality rates in treated bedrooms exceeded 90% for at least 2 weeks, and more than 80% (89.2; 95% CI: 79.98-98.35) for 3 weeks or more. Mortality rates in control houses were zero. Results demonstrate that the immediate impact of TIRS can be delivered by one person using existing products, at an estimated cost for the average household in Mexico of under US$3 per month. Triggered by early outbreak signs, dissemination via community hubs and mass/social media of instructions to treat the home immediately, with monthly re-treatment thereafter, provides a simple means to engage and empower householders. Compatible with integrated vector management strategies, it enables self-protection even if existing agencies falter, a situation exemplified by the potential impact on vector control of the restrictions imposed during the 2020 COVID-19 pandemic.

Field Efficacy Trials of Aerial Ultra-Low-Volume Application of Insecticides Against Caged Aedes aegypti in Mexico.

We evaluated the efficacy of aerial ultra-low-volume (ULV) insecticide spraying in field bioassays with caged Aedes aegypti in May 2017 in Puerto Vallarta, Jalisco, Mexico. The insecticides tested included an organophosphate (Mosquitocida UNO ULV) and a neonicotinoid-pyrethroid combination (Cielo). Two Ae. aegypti populations were evaluated: a field pyrethroid-resistant local strain (Puerto Vallarta) and an insecticide-susceptible laboratory strain (New Orleans). Knockdown after 1 h by both products was ≥97.0%, and mortality after 24 h was ≥98% for the susceptible laboratory strain. Knockdown of the local Puerto Vallarta field strain by both products after 1 h was ≥96.5%; and mosquito mortality after 24 h was also very high (≥98%). Meteorological conditions during this evaluation were favorable for aerial mosquito control and represented conditions that typically occur during adulticide space spray applications. Temperature oscillated between 24°C and 26°C with winds between 6 and 10 km/h. The majority of droplets met the droplet distribution criteria required for the insecticides. The evaluation demonstrated an acceptable performance of both products for Ae. aegypti control when applied undiluted at a rate of 199.4 ml/ha and 73.07 ml/ha for Mosquitocida UNO ULV and Cielo, respectively. The volume median diameter (VMD) droplet size was characterized at 31.3 µm and 37.3 µm, respectively.

Entomological Efficacy of Aerial Ultra-Low Volume Insecticide Applications Against Aedes aegypti (Diptera: Culicidae) in Mexico.

A cluster-randomized controlled trial quantified the entomological efficacy of aerial ultra-low volume (AULV) applications of the insecticide chlorpyrifos against Aedes aegypti in Puerto Vallarta, México, during November-October 2017. The trial involved 16 large (1 × 1 km) clusters distributed between treatment-control arms. Primary endpoint was the abundance of Ae. aegypti indoors (total adults, females, and blood-fed females) collected using Prokopack aspirators. After four consecutive weekly cycles of AULV, all adult Ae. aegypti infestation indices were significantly lower in the treatment arm (OR and IRR ≤ 0.28). Efficacy in reducing indoor Ae. aegypti increased with each weekly application cycle from 30 to 73% (total adults), 33 to 76% (females), and 45.5 to 89% (blood-fed females). Entomological indices remained significantly lower in the treatment arm up to 2 wk after the fourth spraying round. Performing AULV spraying can have significant and lasting entomological impact on Ae. aegypti as long as multiple (ideally four) spray cycles are implemented using an effective insecticide.

Estimating absolute indoor density of Aedes aegypti using removal sampling.

BACKGROUND: Quantification of adult Aedes aegypti abundance indoors has relied on estimates of relative density (e.g. number of adults per unit of sampling or time), most commonly using traps or timed collections using aspirators. The lack of estimates of the sensitivity of collections and lack of a numerical association between relative and the absolute density of adult Ae. aegypti represent a significant gap in vector surveillance. Here, we describe the use of sequential removal sampling to estimate absolute numbers of indoor resting Ae. aegypti and to calculate calibration coefficients for timed Prokopack aspirator collections in the city of Merida, Yucatan State, Mexico. The study was performed in 200 houses that were selected based on recent occurrence of Aedes-borne viral illness in residents. Removal sampling occurred in 10-minute sampling rounds performed sequentially until no Ae. aegypti adult was collected for 3 hours or over 2 consecutive 10-minute periods. RESULTS: A total of 3439 Ae. aegypti were collected. The sensitivity of detection of positive houses in the first sampling round was 82.5% for any adult Ae. aegypti, 78.5% for females, 75.5% for males and 73.3% for blood-fed females. The total number of Ae. aegypti per house was on average ~5 times higher than numbers collected for the first sampling round. There was a positive linear relationship between the relative density of Ae. aegypti collected during the first 10-min round and the absolute density for all adult metrics. Coefficients from the linear regression were used to calibrate numbers from 10-min collections into estimates of absolute indoor Ae. aegypti density for all adults, females and males. CONCLUSIONS: Exhaustive removal sampling represents a promising method for quantification of absolute indoor Ae. aegypti density, leading to improved entomological estimates of mosquito distribution, a key measure in the assessments of the risk pathogen transmission, disease modeling and the evaluation of vector control interventions.

Efficacy of novel indoor residual spraying methods targeting pyrethroid-resistant Aedes aegypti within experimental houses.

Challenges in maintaining high effectiveness of classic vector control in urban areas has renewed the interest in indoor residual spraying (IRS) as a promising approach for Aedes-borne disease prevention. While IRS has many benefits, application time and intrusive indoor applications make its scalability in urban areas difficult. Modifying IRS to account for Ae. aegypti resting behavior, named targeted IRS (TIRS, spraying walls below 1.5 m and under furniture) can reduce application time; however, an untested assumption is that modifications to IRS will not negatively impact entomological efficacy. We conducted a comparative experimental study evaluating the residual efficacy of classically-applied IRS (as developed for malaria control) compared to two TIRS application methods using a carbamate insecticide against a pyrethroid-resistant, field-derived Ae. aegypti strain. We performed our study within a novel experimental house setting (n = 9 houses) located in Merida (Mexico), with similar layouts and standardized contents. Classic IRS application (insecticide applied to full walls and under furniture) was compared to: a) TIRS: insecticide applied to walls below 1.5 m and under furniture, and b) Resting Site TIRS (RS-TIRS): insecticide applied only under furniture. Mosquito mortality was measured eight times post-application (out to six months post-application) by releasing 100 Ae. aegypti females /house and collecting live and dead individuals after 24 hrs exposure. Compared to Classic IRS, TIRS and RS-TIRS took less time to apply (31% and 82% reduction, respectively) and used less insecticide (38% and 85% reduction, respectively). Mortality of pyrethroid-resistant Ae. aegypti did not significantly differ among the three IRS application methods up to two months post application, and did not significantly differ between Classic IRS and TIRS up to four months post application. These data illustrate that optimizing IRS to more efficiently target Ae. aegypti can both reduce application time and insecticide volume with no apparent reduction in entomological efficacy.

House screening with insecticide-treated netting provides sustained reductions in domestic populations of Aedes aegypti in Merida, Mexico.

BACKGROUND: There is a need for effective methods to control Aedes aegypti and prevent the transmission of dengue, chikungunya, yellow fever and Zika viruses. Insecticide treated screening (ITS) is a promising approach, particularly as it targets adult mosquitoes to reduce human-mosquito contact. METHODOLOGY/PRINCIPAL FINDINGS: A cluster-randomised controlled trial evaluated the entomological efficacy of ITS based intervention, which consisted of the installation of pyrethroid-impregnated long-lasting insecticide-treated netting material fixed as framed screens on external doors and windows. A total of 10 treatment and 10 control clusters (100 houses/cluster) were distributed throughout the city of Merida, Mexico. Cross-sectional entomological surveys quantified indoor adult mosquito infestation at baseline (pre-intervention) and throughout four post-intervention (PI) surveys spaced at 6-month intervals corresponding to dry/rainy seasons over two years (2012-2014). A total of 844 households from intervention clusters (86% coverage) were protected with ITS at the start of the trial. Significant reductions in the indoor presence and abundance of Ae. aegypti adults (OR = 0.48 and IRR = 0.45, P<0.05 respectively) and the indoor presence and abundance of Ae. aegypti female mosquitoes (OR = 0.47 and IRR = 0.44, P<0.05 respectively) were detected in intervention clusters compared to controls. This high level of protective effect was sustained for up to 24 months PI. Insecticidal activity of the ITS material declined with time, with ~70% mortality being demonstrated in susceptible mosquito cohorts up to 24 months after installation. CONCLUSIONS/SIGNIFICANCE: The strong and sustained entomological impact observed in this study demonstrates the potential of house screening as a feasible, alternative approach to a sustained long-term impact on household infestations of Ae. aegypti. Larger trials quantifying the effectiveness of ITS on epidemiological endpoints are warranted and therefore recommended.

Deltamethrin resistance in Aedes aegypti results in treatment failure in Merida, Mexico.

The operational impact of deltamethrin resistance on the efficacy of indoor insecticide applications to control Aedes aegypti was evaluated in Merida, Mexico. A randomized controlled trial quantified the efficacy of indoor residual spraying (IRS) against adult Ae. aegypti in houses treated with either deltamethrin (to which local Ae. aegypti expressed a high degree of resistance) or bendiocarb (to which local Ae. aegypti were fully susceptible) as compared to untreated control houses. All adult Ae. aegypti infestation indices during 3 months post-spraying were significantly lower in houses treated with bendiocarb compared to untreated houses (odds ratio <0.75; incidence rate ratio < 0.65) whereas no statistically significant difference was detected between the untreated and the deltamethrin-treated houses. On average, bendiocarb spraying reduced Ae. aegypti abundance by 60% during a 3-month period. Results demonstrate that vector control efficacy can be significantly compromised when the insecticide resistance status of Ae. aegypti populations is not taken into consideration.

Molecular evidence of Rickettsia typhi infection in dogs from a rural community in Yucatán, México.

INTRODUCTION: Rickettsia typhi causes murine or endemic typhus, which is transmitted to humans primarily through flea bites contaminated with feces. Synanthropic and domestic animals also contribute to the infection cycle of R. typhi. Cases of murine typhus in humans were reported in the rural community of Bolmay, Yucatán, México, between 2007 and 2010.  OBJECTIVE: To identify the presence of R. typhi and estimate the frequency of infection in dogs from Bolmay, México, a locality with previous reports of murine typhus in humans.  MATERIALS AND METHODS: Whole blood samples were taken from 128 dogs. Total DNA was extracted for use in the polymerase chain reaction (PCR) to amplify fragments of the 17 kDa and omp B genes and confirms the presence of Rickettsia spp. The reaction products were sequenced, and alignment analysis was performed using the BLAST tool.  RESULTS: The frequency of R. typhi infection in dogs was 5.5 % (7/128). The alignment identified 99% and 100% homology to the R. typhi 17 kDa and omp B genes, respectively.  CONCLUSION: We confirmed the presence of R. typhi in dogs in the studied community but at a low frequency. However, there is potential risk of transmission to humans.

[Effectiveness of commercial repellents against Aedes aegypti (L.) in Yucatan, México]. / Efectividad de repelentes comerciales disponibles contra el mosquito Aedes aegypti (L.) en Yucatán, México.

OBJECTIVE: We assessed the efficacy of commercial repellents available in Yucatan against Aedes aegypti, vector of dengue, Chikungunya and Zika. MATERIALS AND METHODS: Protection time was determined based on WHO/CTD/ WHOPES/IC y la NOM-032-SSA2-20I4. RESULTS: Products with DEET (N, N-diethyl-3-methylbenzamide) at 25% met the recommended protection. Efficacy was directly proportional to the concentration of DEET; botanicals repellents resulted no protective. CONCLUSIONS: Repellents with DEET provided more protection against Ae. aegypti and botanical repellents, including impregnated wristbands, provided no protection.

Efectividad de repelentes comerciales disponibles contra el mosquito Aedes aegypti (L ) en Yucatán, México / Effectiveness of commercial repellents against Aedes aegypti (L ) in Yucatan, México

Resumen: Objetivo: Determinar la eficacia de repelentes comerciales disponibles en Yucatán contra el mosquito Aedes aegypti, vector del dengue, Chikungunya y Zika. Material y métodos: Se determinó el tiempo de protección con base en el protocolo WHO/CTD/WHOPES/IC y la NOM-032-SSA2-2014. Resultados: Sólo el repelente con DEET (N, N-dietil-3-metilbenzamida) al 25% cumplió con la protección recomendada. La eficacia fue directamente proporcional a la concentración del DEET; aquéllos con componentes botánicos fueron poco o nada protectores. Conclusiones: Los resultados muestran que los repelentes con DEET proveen protección contra Ae. Aegypti; los repelentes botánicos, incluyendo las pulseras impregnadas, ofrecen nula protección.

Abstract: Objective: We assessed the efficacy of commercial repellents available in Yucatan against Aedes aegypti, vector of dengue, Chikungunya and Zika. Materials and methods: Protection time was determined based on WHO/CTD/ WHOPES/IC y la NOM-032-SSA2-20I4. Results: Products with DEET (N, N-diethyl-3-methylbenzamide) at 25% met the recommended protection. Efficacy was directly proportional to the concentration of DEET; botanicals repellents resulted no protective. Conclusions: Repellents with DEET provided more protection against Ae. aegypti and botanical repellents, including impregnated wristbands, provided no protection.