Mosquito Collections Research Articles

Cattle-related risk factors for malaria in southwest Ethiopia: a cross-sectional study

BackgroundDespite the low to moderate intensity of malaria transmission present in Ethiopia, malaria is still a leading public health problem. Current vector control interventions, principally long-lasting insecticidal nets and indoor residual spraying, when deployed alone or in combination, are insufficient to control the dominant vector species due to their exophagic and exophilic tendencies. Zooprophylaxis presents a potential supplementary vector control method for malaria; however, supporting evidence for its efficacy has been mixed.MethodsTo identify risk factors of malaria and to estimate the association between cattle and Anopheles vector abundance as well as malaria risk, a cross-sectional study was conducted in a village near Arba Minch, Ethiopia. Epidemiological surveys (households = 95, individuals = 463), mosquito collections using CDC light traps and a census of cattle and human populations were conducted. To capture environmental conditions, land cover and water bodies were mapped using satellite imagery. Risk factor analyses were performed through logistic, Poisson, negative binomial, and spatial weighted regression models.ResultsThe only risk factor associated with self-reported malaria illness at an individual level was being a child aged 5 or under, where they had three times higher odds than adults. At the household level, variables associated with malaria vector abundance, especially those indoors, included socioeconomic status, the proportion of children in a household and cattle population density.ConclusionsStudy results are limited by the low abundance of malaria vectors found and use of self-reported malaria incidence. Environmental factors together with a household’s socioeconomic status and host availability played important roles in the risk of malaria infection in southwest Ethiopia. Cattle abundance in the form of higher cattle to human ratios may act as a protective factor against mosquito infestation and malaria risk. Humans should remain indoors to maximize potential protection against vectors and cattle kept outside of homes.

Effects of Indoor Environmental Factors and House Structures on Vaporization of Active Ingredient from Spatial Repellent Devices in Rural Houses in Malawi.

The use of a metofluthrin-impregnated spatial repellent device (MSRD) is a new and effective method for preventing mosquito blood feeding. Indoor environmental factors such as room temperature and ventilation rate are thought to be important for MSRD activity. Measurements of room temperature and vaporization of metofluthrin from MSRD in typical rural metal-roof and thatched-roof houses in southeastern Malawi were conducted. The relationship between house structure and the number of collected Anopheline mosquitoes with and without MSRD treatment was also investigated. The difference between daytime and nighttime room temperature was significantly higher in metal-roof houses than in thatched-roof houses. The vaporization of metofluthrin from the MSRD was not accelerated by the high room temperature, but by the high indoor air flow by ventilation. The number of mosquito collections was significantly higher in thatched-roof houses than in metal-roof houses. MSRD-treated thatched-roof houses have a higher probability of mosquito infestation, but the vaporization of metofluthrin is also higher because of indoor air flow, resulting in a reduction in mosquito numbers. Metal-roof houses with closed eaves reduce the probability of mosquito invasion, and a longer predicted effectiveness occurs with MSRD because of the controlled release of metofluthrin through lower indoor air flow.

Efficacy of Trapping Methods in the Collection of Eretmapodites (Diptera: Culicidae) Mosquitoes in an Afrotropical Rainforest Region, South western Cameroon.

Very little data exist on the biology of an afrotropical rainforest mosquito Eretmapodites (Er.) in a world undergoing dramatic changes due to deforestation. The aim was to assess the efficacy of different trapping methods in the collection of Er. mosquito in forested area. This was a longitudinal study involving collection of mosquitoes for over two years. Multiple collection methods (grouped into two categories), were used; i) net baited and un-baited traps to collect adults, ii) techniques that target immature stages subsequently reared to adults. All males were identified by genitalia dissection. Five thousand seven hundred and four mosquitoes representing 11 genera among which 2,334 Er. were identified. Mosquito abundance was highest in the net traps (n = 1276 (56.4%)) and sweep nets (n = 393(17.4%)) respectively. The abundance was highest in green colored net traps (435(34.09%)) with significant value of χ2= 40.000, P < 0.001 and in pigeons baited traps (473 (37.06%)) with significant value of χ2= 42.000, P = 0.003. The diversity ranges from H' = 2.65; DS = 0.84; SR = 24; ACE = 24.77 in sweep net to H' = 0; DS = 0; SR = 1; ACE = 1 in rock pool among males mosquitoes. While for females, H = 1.14; DS = 0.71; SR = 5; ACE = 5.16, in sweep net to H = 0; DS = 0; SR = 1; ACE = 1 in rock pool, tarpaulin, resting cage. Net traps, bamboo pot, and sweep netting are efficient in collecting high abundance of forest mosquitoes in the Talanagaye rainforest.

Diurnal biting of malaria mosquitoes in the Central African Republic indicates residual transmission may be “out of control”

Malaria control interventions target nocturnal feeding of the Anopheles vectors indoors to reduce parasite transmission. Mass deployment of insecticidal bed nets and indoor residual spraying with insecticides, however, may induce mosquitoes to blood-feed at places and at times when humans are not protected. These changes can set a ceiling to the efficacy of these control interventions, resulting in residual malaria transmission. Despite its relevance for disease transmission, the daily rhythmicity of Anopheles biting behavior is poorly documented, most investigations focusing on crepuscular hours and nighttime. By performing mosquito collections 48-h around the clock, both indoors and outdoors, and by modeling biting events using circular statistics, we evaluated the full daily rhythmicity of biting in urban Bangui, Central African Republic. While the bulk of biting by Anopheles gambiae, Anopheles coluzzii, Anopheles funestus, and Anopheles pharoensis occurred from sunset to sunrise outdoors, unexpectedly ∼20 to 30% of indoor biting occurred during daytime. As biting events did not fully conform to any family of circular distributions, we fitted mixtures of von Mises distributions and found that observations were consistent with three compartments, corresponding indoors to populations of early-night, late-night, and daytime-biting events. It is not known whether these populations of biting events correspond to spatiotemporal heterogeneities or also to distinct mosquito genotypes/phenotypes belonging consistently to each compartment. Prevalence of Plasmodium falciparum in nighttime- and daytime-biting mosquitoes was the same. As >50% of biting occurs in Bangui when people are unprotected, malaria control interventions outside the domiciliary environment should be envisaged.

Breeding Preference and Bionomics of Anopheles spp. at the Malarial Endemic Area, Runut Village, East Nusa Tenggara Province, Indonesia

Introduction: Anopheles mosquito is transmitting malaria, one of the health problems in Indonesia. Understanding Anopheles mosquito behaviour and its breeding preference is one of the crucial keys to prevent malaria transmission. This study aimed to identify the breeding place distribution and bionomics of Anopheles spp. in Runut village, Sikka district, East Nusa Tenggara. Methods: A descriptive observational study was conducted in Runut village, Waigete sub- district, in April 2018. Anopheles spp. larvae were collected in 7 suspected breeding places consisting of 2 rice fields, 3 fish ponds, and 2 puddles. Mosquitos behaviour was observed using bed-net traps located inside and outside the house from 7.15 PM to 1.15 AM after obtaining informed consent. Mosquito collection using bed-net trap were performed for 40 minutes then followed by resting mosquito collection for 10 minutes. Results: Anopheles spp. larvae were found in most of the suspected aquatic habitats, presenting different densities and together with larvae of the other mosquito species. Relatively high number of Anopheles spp. larvae was obtained from a puddle. Only one female mosquito of Anopheles spp. resting on the wall inside house was found around 00.55 – 01.05 AM and resulted in low mosquito density determination. Conclusion: Anopheles spp. larvae were harbouring in most of the aquatic habitats and one puddle contained moderately abundant larvae of Anopheles in Runut village, Sikka district, East Nusa Tenggara, Indonesia. Even only one Anopheles spp. mosquito was detected inside the house, residents in Runut village should regular use insecticide-treated bed nets and continuous observation of mosquito breeding places especially puddles to prevent malaria and other mosquito-borne diseases.

Beyond eves and cracks: An interdisciplinary study of socio-spatial variation in urban malaria transmission in Ethiopia.

During the past century, the global trend of reduced malaria transmission has been concurrent with increasing urbanization. Although urbanization has traditionally been considered beneficial for vector control, the adaptation of malaria vectors to urban environments has created concerns among scientific communities and national vector control programs. Since urbanization rates in Ethiopia are among the highest in the world, the Ethiopian government developed an initiative focused on building multi-storied units organized in condominium housing. This study aimed to develop an interdisciplinary methodological approach that integrates architecture, landscape urbanism, medical anthropology, and entomology to characterize exposure to malaria vectors in this form of housing in three condominiums in Jimma Town. Mosquitoes were collected using light trap catches (LTCs) both indoor and outdoor during 2019's rainy season. Architectural drawings and ethnographic research were superposed to entomological data to detect critical interactions between uses of the space and settlement conditions potentially affecting malaria vector abundance and distribution. A total of 34 anopheline mosquitoes comprising three species (Anopheles gambiae s.l, An. pharoensis and An. coustani complex) were collected during the three months of mosquito collection. Anopheles gambiae s.l, the principal malaria vector in Ethiopia, was the predominant species of all the anophelines collected. Distribution of mosquito breeding sites across scales (household, settlement, urban landscape) is explained by environmental conditions, socio-cultural practices involving modification of existing spaces, and systemic misfits between built environment and territory. Variations in mosquito abundance and distribution in this study were mainly related to standard building practices that ignore the original logics of the territory, deficiency of water and waste disposal management systems, and adaptations of the space to fit heterogeneous lifestyles of residents. Our results indicate that contextualizing malaria control strategies in relation to vector ecology, social dynamics determining specific uses of the space, as well as building and territorial conditions could strengthen current elimination efforts. Although individual housing remains a critical unit of research for vector control interventions, this study demonstrates the importance of studying housing settlements at communal level to capture systemic interactions impacting transmission at the household level and in outdoor areas.

Comparison of Different Sampling Methods to Catch Lymphatic Filariasis Vectors in a Sudan Savannah Area of Mali.

There is a need for better tools to monitor the transmission of lymphatic filariasis and malaria in areas undergoing interventions to interrupt transmission. Therefore, mosquito collection methods other than human landing catch (HLC) are needed. This study aimed to compare the Ifakara tent trap type C (ITTC) and the Biogents sentinel trap (BGST) to the HLC in areas with different vector densities. Mosquitoes were collected in two villages in Mali from July to December in 2011 and 2012. The three methods were implemented at each site with one ITTC, one BGST, and one HLC unit that consisted of one room with two collectors-one indoor and the other outdoor. The Anopheles collected in 2011 were individually dissected, whereas those from 2012 were screened in pools using reverse transcription-polymerase chain reaction (RT-PCR) to determine the maximum infection prevalence likelihood (MIPL) for Wuchereria bancrofti and Plasmodium falciparum. The dissection of the females also allowed to assess the parity rates, as well its results. Over the 2 years, the HLC method collected 1,019 Anopheles, yields that were 34- and 1.5-fold higher than those with the BGST and ITTC, respectively. None of the dissected Anopheles were infected. The RT-PCR results showed comparable MIPL between HLC and ITTC for W. bancrofti with one infected pool from each trap's yield (respectively 0.03% [0.0009-0.2%] and 0.04% [0.001-0.2%]). For P. falciparum, no infected pool was recovered from BGST. The ITTC is a good alternative to HLC for xenomonitoring of program activities.

Evaluation of the protective efficacy of a spatial repellent to reduce malaria incidence in children in Mali compared to placebo: study protocol for a cluster-randomized double-blinded control trial (the AEGIS program)

BackgroundSpatial repellents have been widely used for the prevention of mosquito bites but their efficacy in reducing mosquito-borne diseases has never been evaluated in Africa. Additionally, spatial repellents have the potential of being critical tools in the prevention of mosquito-borne diseases in contexts where typical vectors control efforts such as insecticide-treated nets (ITNs) and indoor residual spray (IRS) are inaccessible or underutilized such as among displaced populations or in emergency relief settings. To address this knowledge gap, Kolondieba District, Sikasso Region, Mali was selected as a site to estimate the impact of the Mosquito Shield™, a spatial repellent that incorporates transfluthrin on a plastic sheet, on malaria-related outcomes. Over the past decade, the Region of Sikasso, Health districts of Kadiolo, Yorosso, and Kolondieba have remained among the most afflicted, characterized by an annual parasite incidence of more than 116 cases per 1000 population [1] and a Plasmodium falciparum prevalence rate of 29.7% [2].MethodsCluster-randomized, placebo-controlled, double-blinded clinical trial, whereby children ≥ 6 months to < 10 years old will be enrolled and followed to determine the time to malaria infection with monthly blood samples for microscopic diagnosis. A total of 1920 subjects (HHs) will be enrolled in 60 clusters (30 spatial repellent, 30 placebo). Malaria incidence will be estimated and compared to demonstrate and quantify the protective efficacy (PE) of a spatial repellent, in reducing malaria infection. Monthly mosquito collections using CDC light traps will be conducted to determine if there are entomological correlates of spatial repellent efficacy that may be useful for the evaluation of new spatial repellents. Quarterly human landing catches (HLC) will assess the behavioral effects of the intervention.DiscussionFindings will serve as an efficacy trial of spatial repellent products for sub-Saharan Africa. Findings will be submitted to the World Health Organization Vector Control Advisory Group (WHO VCAG) for assessment of whether spatial repellents have “public health value.” Entomological outcomes will also be measured as proxies of malaria transmission to help develop guidelines for the evaluation of future spatial repellent products.Trial registrationClinicalTrials.govNCT04795648. Registered on March 12, 2021.

Continuing the Role of the Citizen Scientist

New introductions to invasive mosquitoes, Aedes aegypti and Aedes albopictus, increase the risk for vector-borne diseases such as dengue, chikungunya, and Zika in the United States. Tracking these new introductions is more important than ever. This lesson plan focuses on the collection of mosquito larvae and pupae before the onset of summer with a focus on insect development. Students will observe the immature mosquitoes grow and metamorphose into adults. Novel aspects include collecting larvae and pupae around the home/school, observing the mosquito life cycle by safely rearing them from immature stages (larvae and pupae) to adults, and learning important background information on mosquito biology and pathogens that mosquitoes can transmit. The lesson describes new tools to use with the Invasive Mosquito Project, an international citizen science–based mosquito surveillance program exploring mosquitoes and the pathogens they may transmit to interested community members (including students and teachers) and their companion animals. This project is a stand-alone or follow-up lesson plan to the mosquito egg collection lesson used prior to the onset of winter (see Cohnstaedt et al., 2016). Students are able to participate in a hands-on tutorial to build a sealed emergence chamber to safely raise mosquitoes. Long-term data sets can also be used by teachers and students for further classroom discussions on the risks of mosquito-borne illnesses nationwide. This lesson further focuses on how individuals must play an active role in protecting their communities and pets from illness and increasing awareness of the dangerous pathogens mosquitoes can transmit and the importance of mosquito management. Materials from this lesson plan (available at http://www.citizenscience.us) can be adapted for each classroom but are best-suited for middle school to high school classes, as well as Advanced Placement classes.

Diel activity patterns of two distinct populations of Aedes aegypti in Miami, FL and Brownsville, TX

The diel biting activity of Aedes (Stegomyia) aegypti (L) populations was extensively investigated in the early 1900s to gain more information on the biology of Ae. aegypti, and this information was used to devise effective approaches to controlling populations of this species and protect the human population from widespread arbovirus outbreaks. However, few contemporary studies are available regarding the diel activity patterns of Ae. aegypti. To assess the diel activity patterns of Ae. aegypti in southern Florida and Texas, we conducted 96-h uninterrupted mosquito collections once each month from May through November 2019 in Miami, Florida, and Brownsville, Texas, using BG-Sentinel 2 Traps. The overall diel activity pattern in both cities was bimodal with morning and evening peak activity between 7:00 and 8:00 and between 19:00 and 20:00. There were significant daily, monthly, seasonal, and site-specific differences in activity patterns, but these differences did not affect the overall peak activity times. These differences suggest daily, monthly, seasonal, and site-specific variations in human exposure to Ae. aegypti. Our observations can be used in planning and executing Ae. aegypti vector control activities in southern Florida and southern Texas, specifically those targeting the adult mosquito populations.

House design and risk of malaria, acute respiratory infection and gastrointestinal illness in Uganda: A cohort study.

House construction is rapidly modernizing across Africa but the potential benefits for human health are poorly understood. We hypothesised that improvements to housing would be associated with reductions in malaria, acute respiratory infection (ARI) and gastrointestinal illness in an area of low malaria endemicity in Uganda. Data were analysed from a cohort study of male and female child and adult residents (n = 531) of 80 randomly-selected households in Nagongera sub-county, followed for 24 months (October 4, 2017 to October 31, 2019). Houses were classified as modern (brick walls, metal roof and closed eaves) or traditional (all other homes). Light trap collections of mosquitoes were done every two weeks in all sleeping rooms. Every four weeks, we measured malaria infection (using microscopy and qPCR to detect malaria parasites), incidence of malaria, ARI and gastrointestinal illness. We collected 15,780 adult female Anopheles over 7,631 nights. We collected 13,277 blood samples of which 10.2% (1,347) were positive for malaria parasites. Over 958 person years we diagnosed 38 episodes of uncomplicated malaria (incidence 0.04 episodes per person-year at risk), 2,553 episodes of ARI (incidence 2.7 episodes per person-year) and 387 episodes of gastrointestinal illness (incidence 0.4 episodes per person-year). Modern houses were associated with a 53% lower human biting rate compared to traditional houses (adjusted incidence rate ratio [aIRR] 0.47, 95% confidence interval [CI] 0.32-0.67, p<0.001) and a 24% lower incidence of gastrointestinal illness (aIRR 0.76, 95% CI 0.59-0.98, p = 0.04) but no changes in malaria prevalence, malaria incidence nor ARI incidence. House improvements may reduce mosquito-biting rates and gastrointestinal illness among children and adults. For the health sector to leverage Africa's housing modernization, research is urgently needed to identify the healthiest house designs and to assess their effectiveness across a range of epidemiological settings in sub-Saharan Africa.

The effect of light and ventilation on house entry by Anopheles arabiensis sampled using light traps in Tanzania: an experimental hut study

BackgroundIn sub-Saharan Africa, house design and ventilation affects the number of malaria mosquito vectors entering houses. This study hypothesized that indoor light from a CDC-light trap, visible from outside a hut, would increase entry of Anopheles arabiensis, an important malaria vector, and examined whether ventilation modifies this effect.MethodsFour inhabited experimental huts, each situated within a large chamber, were used to assess how light and ventilation affect the number of hut-entering mosquitoes in Tanzania. Each night, 300 female laboratory-reared An. arabiensis were released inside each chamber for 72 nights. Nightly mosquito collections were made using light traps placed indoors. Temperature and carbon dioxide concentrations were measured using data loggers. Treatments and sleepers were rotated between huts using a randomized block design.ResultsWhen indoor light was visible outside the huts, there was an 84% increase in the odds of collecting mosquitoes indoors (Odds ratio, OR = 1.84, 95% confidence intervals, 95% CI 1.74–1.95, p < 0.001) compared with when it was not. Although the odds of collecting mosquitoes in huts with closed eaves (OR = 0.54, 95% CI 0.41–0.72, p < 0.001) was less than those with open eaves, few mosquitoes entered either type of well-ventilated hut. The odds of collecting mosquitoes was 99% less in well-ventilated huts, compared with poorly-ventilated traditional huts (OR = 0.01, 95% CI 0.01–0.03, p < 0.001). In well-ventilated huts, indoor temperatures were 1.3 °C (95% CI 0.9–1.7, p < 0.001) cooler, with lower carbon dioxide (CO2) levels (mean difference = 97 ppm, 77.8–116.2, p < 0.001) than in poorly-ventilated huts.ConclusionAlthough light visible from outside a hut increased mosquito house entry, good natural ventilation reduces indoor carbon dioxide concentrations, a major mosquito attractant, thereby reducing mosquito-hut entry.

Larval habitats, species composition and distribution of malaria vectors in regions with autochthonous and imported malaria in Roraima state, Brazil

BackgroundMalaria control requires local action. Assessing the vector diversity and abundance provides information on the local malariogenic potential or risk of transmission. This study aimed to determine the Anopheles species composition, habitats, seasonal occurrence, and distribution in areas with autochthonous and imported malaria cases in Roraima State.MethodsA longitudinal study was conducted from January 2017 to October 2018, sampling larvae and adult mosquitoes in three municipalities of Roraima State: Boa Vista, Pacaraima and São João da Baliza. These areas have different risks of malaria importation. Four to six mosquito larval habitats were selected for larval sampling at each municipality, along with two additional sites for adult mosquito collection. All larval habitats were surveyed every two months using a standardized larval sampling methodology and MosqTent for adult mosquitoes.ResultsA total of 544 Anopheles larvae and 1488 adult mosquitoes were collected from the three municipalities studied. Although the species abundance differed between municipalities, the larvae of Anopheles albitarsis s.l., Anopheles nuneztovari s.l. and Anopheles triannulatus s.l. were collected from all larval habitats studied while Anopheles darlingi were collected only from Boa Vista and São João da Baliza. Adults of 11 species of the genus Anopheles were collected, and the predominant species in Boa Vista was An. albitarsis (88.2%) followed by An. darlingi (6.9%), while in São João da Baliza, An. darlingi (85.6%) was the most predominant species followed by An. albitarsis s.l. (9.2%). In contrast, the most abundant species in Pacaraima was Anopheles braziliensis (62%), followed by Anopheles peryassui (18%). Overall, the majority of anophelines exhibited greater extradomicile than peridomicile-biting preference. Anopheles darlingi was the only species found indoors. Variability in biting times was observed among species and municipalities.ConclusionThis study revealed the composition of anopheline species and habitats in Boa Vista, Pacaraima and São João da Baliza. The species sampled differed in their behaviour with only An. darlingi being found indoors. Anopheles darlingi appeared to be the most important vector in São João da Baliza, an area of autochthonous malaria, and An. albitarsis s.l. and An. braziliensis in areas of low transmission, although there were increasing reports of imported malaria. Understanding the diversity of vector species and their ecology is essential for designing effective vector control strategies for these municipalities.

Vector composition, abundance, biting patterns and malaria transmission intensity in Madang, Papua New Guinea: assessment after 7\xa0years of an LLIN-based malaria control programme

BackgroundA malaria control programme based on distribution of long-lasting insecticidal bed nets (LLINs) and artemisinin combination therapy began in Papua New Guinea in 2009. After implementation of the programme, substantial reductions in vector abundance and malaria transmission intensity occurred. The research reported here investigated whether these reductions remained after seven years of sustained effort.MethodsAll-night (18:00 to 06:00) mosquito collections were conducted using human landing catches and barrier screen methods in four villages of Madang Province between September 2016 and March 2017. Anopheles species identification and sporozoite infection with Plasmodium vivax and Plasmodium falciparum were determined with molecular methods. Vector composition was expressed as the relative proportion of different species in villages, and vector abundance was quantified as the number of mosquitoes per barrier screen-night and per person-night. Transmission intensity was quantified as the number of sporozoite-infective vector bites per person-night.ResultsFive Anopheles species were present, but vector composition varied greatly among villages. Anopheles koliensis, a strongly anthropophilic species was the most prevalent in Bulal, Matukar and Wasab villages, constituting 63.7–73.8% of all Anopheles, but in Megiar Anopheles farauti was the most prevalent species (97.6%). Vector abundance varied among villages (ranging from 2.8 to 72.3 Anopheles per screen-night and 2.2–31.1 Anopheles per person-night), and spatially within villages. Malaria transmission intensity varied among the villages, with values ranging from 0.03 to 0.5 infective Anopheles bites per person-night. Most (54.1–75.1%) of the Anopheles bites occurred outdoors, with a substantial proportion (25.5–50.8%) occurring before 22:00.ConclusionThe estimates of vector abundance and transmission intensity in the current study were comparable to or higher than estimates in the same villages in 2010–2012, indicating impeded programme effectiveness. Outdoor and early biting behaviours of vectors are some of the likely explanatory factors. Heterogeneity in vector composition, abundance and distribution among and within villages challenge malaria control programmes and must be considered when planning them.

Comparison of Insecticide Resistance and Its Enzyme Mechanisms among &amp;lt;i&amp;gt;Aedes aegypti&amp;lt;/i&amp;gt; Collected with Three Methods in a Dengue-Endemic City in Southern Mexico

Background: Despite the physical and chemical effort to control Aedes aegypti, the arboviruses transmission in the south of Mexico remains latent. Trying to improve the methods of entomological surveillance routinely used, whether the estimation of resistance to insecticides used for its control, as well as their enzyme mechanisms, were influenced by the phase in which the mosquitoes were collected through three different collection methods was investigated. Materials and Methods: Mosquito collections from the “5 de Febrero” neighborhood in Tapachula, Mexico were obtained by ovitraps, larvitraps, and a CDC backpack aspirator. Insecticide resistance of F1 females was determined by WHO diagnostic doses and resistance ratios (RR50), furthermore, levels of insecticide metabolism enzymes were determined by biochemical assays. Results: Overall, in mosquitoes collected by ovitraps, larvitraps, and CDC backpack aspirator respectively, the low mortalities obtained with the discriminant dose to Malathion (27.57%, 26.97%, and 26.91%), and to Bendiocarb (50.5%, 45.36%, and 54.97%) suggest resistance. However, LC50 for Malathion (0.922, 0.934, and 0.915) and for Bendiocarb (0.112, 0.109, and 0.107); and the low resistance ratios (RR50) for Malathion (3.34, 3.29, and 3.27) and for Bendiocarb (2.15, 2.1, and 2.06) does not suggest resistance. Although a slight numerical variation is observed between the three LC50 values, the overlap observed between their confidence intervals allows us to assume that there were no differences between the three methods. In general, esterases (determined with three substrates), glutathion S-transferases (GST) and cytochromes P450 were statistically higher than those of the susceptible strain; and the three enzyme levels were statistically different among the three collection methods (P < 0.01), being those collected with CDC backpack aspirator with the highest levels. Conclusion: Although using a CDC backpack aspirator demonstrated being the best collection method determining a specific resistance mechanism (as elevation at the enzyme level) in the mosquito adult phase, any collection method is reliable to determine whether a field mosquito population is resistant or susceptible to an insecticide.

Malaria vector sibling species distribution in different endemic districts of Punjab, India.

Malaria transmission in Punjab, India is mainly seasonal with variation in its endemicity that may be due to varying vector behaviour in different areas of the state, primarily attributed to the existence of sibling species complexes among the vector species. So far there is no report regarding the existence of malaria vectors sibling species in the state of Punjab, therefore, the present study was planned to investigate the status of sibling species of two main vectors of malaria viz. Anopheles culcifacies and Anopheles fluviatilis in different districts of Punjab. Mosquito collections were made through hand catch in the morning hours. Malaria vector species An. culicifacies and An. fluviatilis were morphologically identified and man hour density was calculated. Both the vector species were subjected to molecular assays for sibling species identification through amplification of D3 domain of 28S ribosomal DNA by allele-specific PCR. Four sibling species of An. culicifacies, were identified viz. A, B, C and E. Species A was identified from Bhatinda district, species B, C and E from. S.A.S. Nagar and species C from Hoshiarpur. Two sibling species S and T of An. fluviatilis were identified from districts S.A.S. Nagar and Rupnagar. Presence of four sibling species of An. culicifacies and two sibling species of An. fluviatilis in Punjab necessitates planning of longitudinal studies to ascertain their role in disease transmission so that appropriate interventions may be applied to achieve malaria elimination.

Impact of full coverage of population with Long Lasting Insecticidal Nets (LLIN) on malaria vector density in Gadchiroli district of Maharashtra

Background & objectives: Long-lasting insecticidal nets (LLINs) are a major weapon in the fight against malaria. But protection of each of family member is not possible with current distribution system with 80% coverage. Therefore, we hypothesized that distribution of LLIN to every member of a household will be more effective at reducing vector density and ultimately malaria in endemic areas like district Gadchiroli of Maharashtra. Methods: The three sub centers Kulbhati, Pannemara and Yerkad of Murumgaon Primary Health Center of Gadchiroli district were selected for the study based on malaria incidence and LLIN distribution. Prestructured questionnaire were filled to know the distribution and use of LLIN as baseline, thereafter additional LLIN were distributed in selected villages for 100% coverage of population. Entomological surveys were undertaken to know the impact of LLIN on vector mosquito density. Results: A total of 250 houses were surveyed in these villages and questionnaires were filled regarding bednet distribution and their use. 507 LLIN were distributed in study villages (population 1270) of Murumgaon PHC. In four selected villages, additional 409 LLIN were distributed for 100% coverage of population with bed nets. Man hour density of vector mosquito in human dwelling in the experimental villages was significantly reduced, 0-19 in comparison to control (0-66). In cattle sheds, MHD of An. culicifacies was observed to be 1-243 in experimental villages vis-ΰ-vis 1-179 in control villages. During exit trap collection of mosquitoes, ~94 % mosquitoes were deterred from blood feeding. Interpretation & conclusion: In this study there was a significant impact of insecticide treated nets on vector mosquito density in human dwellings. Thus 100% coverage of human populations with LLIN may be an important tool in controlling malaria situation in endemic areas like Gadchiroli.

Artificial resting sites: An alternative sampling method for adult mosquitoes.

Mosquito collections are commonly conducted with baited traps predominantly attracting host-seeking females. In contrast, resting sites are generally colonized by a broader range of the mosquito population, including a higher proportion of males and blood-engorged females. This study evaluates the sampling success of different artificial resting sites, attached to a deciduous or coniferous tree at different heights. As standard sampling method, carbon dioxide-baited Biogents Sentinel traps (BG traps) were operated in parallel. BG traps caught a higher number of specimens compared to the resting sites. However, the proportion of blood-engorged females and males was higher in resting sites. More Culiseta spp. specimens were collected in resting sites compared to BG traps, but less Aedes spp. specimens. In general, fewer specimens and species were recorded in small resting sites and at top height level compared to medium or large resting sites at medium or ground level. The proportion of males was highest at the ground, while the proportion of engorged females was highest at medium and top level. Due to the higher proportion of blood-engorged females, artificial resting sites are especially useful for studies of host-feeding patterns or xenosurveillance. Low costs and efforts allow a cost-effective increase of the number of resting sites per sampling site to collect more mosquitoes.

Entomological Investigation Following a Zika Outbreak in Brownsville, Texas.

In November and December 2016, an outbreak of locally transmitted Zika occurred in Brownsville, TX. The Texas Department of State Health Services requested for a Centers for Disease Control and Prevention (CDC) Epi Aid, and as part of that Epi Aid a team of CDC entomologists was deployed in January 2017. The mission was to improve mosquito-based arbovirus surveillance and evaluate the possibility of continuing local Zika virus (ZIKV) transmission in the city. The mosquito-based arbovirus surveillance program was expanded from 4 to 40 BG-Sentinel traps evenly distributed throughout the city. Over a 2-wk period, 15 mosquito species were detected; the most abundant species were Culex quinquefasciatus, Aedes aegypti, and Ae. albopictus, which accounted for 66.7%, 16.2%, and 5.7% of the total mosquito collection, respectively. The relative abundance of Ae. aegypti (1.0 mosquitoes/trap/day) and Ae. albopictus (0.4 mosquitoes/trap/day) was very low and unlikely to initiate and/or sustain ZIKV transmission. Zika virus was not detected in the mosquitoes collected, suggesting no or extremely low ZIKV transmission at that time.

Coverage, Usage, Physical Integrity, and Bio-efficacy of Olyset Nets in the Plateau Region, South Benin Following the 2011 Nationwide Distribution.

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.