Invasive Anopheles stephensi in Africa: insights from Asia.

Anopheles stephensi is a highly competent urban malaria vector species, endemic in South Asia and the Persian Gulf, which has colonised eight countries in sub-Saharan Africa (SSA) since 2013 and is now spreading uncontrollably. In urban areas of Africa, where malaria transmission has previously been low or non-existent, the invasion of An. stephensi represents a significant problem, particularly to immunologically naïve populations. Despite this rapidly advancing threat, there is a paucity of information regarding the bionomics of An. stephensi in SSA. Here, we offer a critical synthesis of literature from An. stephensi's native range, focusing on the future of An. stephensi in a rapidly urbanising Africa, and highlighting key questions that warrant prioritisation by the global malaria vector control community.

Malaria Vector Bionomics: Countrywide Surveillance Study on Implications for Malaria Elimination in India.

BACKGROUND: The biological characteristics of mosquito vectors vary, impacting their response to control measures. Thus, having up-to-date information on vector bionomics is essential to maintain the effectiveness of existing control strategies and tools, particularly as India aims for malaria elimination by 2030. OBJECTIVE: This study aims to assess the proportions of vector species resting indoors and outdoors, determine their preference for host biting/feeding, identify transmission sites, and evaluate the susceptibility of vectors to insecticides used in public health programs. METHODS: Mosquito collections were conducted in 13 districts across 8 Indian states from 2017 to 2020 using various methods to estimate their densities. Following morphological identification in the field, sibling species of Anopheles mosquitoes were identified molecularly using polymerase chain reaction (PCR)-specific alleles. Plasmodium falciparum and Plasmodium vivax infections in the vectors were detected using enzyme-linked immunosorbent assay (ELISA) and PCR assays. In addition, we assessed the insecticide susceptibility status of primary malaria vectors following the World Health Organization (WHO) protocol. RESULTS: Anopheles culicifacies, a primary malaria vector, was collected (with a man-hour density ranging from 3.1 to 15.9) from all states of India except those in the northeastern region. Anopheles fluviatilis, another primary vector, was collected from the states of Madhya Pradesh, Maharashtra, Karnataka, and Odisha. In Haryana and Karnataka, An. culicifacies sibling species A predominated, whereas species C and E were predominant in Madhya Pradesh and Maharashtra. An. culicifacies displayed mainly endophilic behavior across all states, except in Madhya Pradesh, where the proportion of semigravid and gravid mosquitoes was nearly half of that of unfed mosquitoes. The human blood index of An. culicifacies ranged from 0.001 to 0.220 across all study sites. The sporozoite rate of An. culicifacies ranged from 0.06 to 4.24, except in Madhya Pradesh, where none of the vector mosquitoes were found to be infected with the Plasmodium parasite. In the study area, An. culicifacies exhibited resistance to DDT (dichlorodiphenyltrichloroethane; with <39% mortality). Moreover, it showed resistance to malathion (with mortality rates ranging from 49% to 78%) in all districts except Angul in Odisha and Palwal in Haryana. In addition, resistance to deltamethrin was observed in districts of Maharashtra, Gujarat, Haryana, and Karnataka. CONCLUSIONS: Our study offers vital insights into the prevalence, resting behavior, and sibling species composition of malaria vectors in India. It is evident from our findings that resistance development in An. culicifacies, the primary vector, to synthetic pyrethroids is on the rise in the country. Furthermore, the results of our study suggest a potential change in the resting behavior of An. culicifacies in Madhya Pradesh, although further studies are required to confirm this shift definitively. These findings are essential for the development of effective vector control strategies in India, aligning with the goal of malaria elimination by 2030.

A Taxonomic Revision of the Genus Cleopus Dejean, 1821 (Coleoptera, Curculionidae), with Descriptions of 13 New Species.

The genus Cleopus Dejean, 1821 is herein revised for the first time. Based on adult morphological characteristics, 18 species are recognized as valid. Thirteen species, all distributed in the Eastern Palaearctis or Oriental region, are described as new: C. aduncirostris sp. n.; C. cognatus sp. n.; C. confusus sp. n.; C. dohertyi sp. n.; C. hajeki sp. n.; C. lirenae sp. n.; C. longitarsis sp. n.; C. minutus sp. n.; C. pallidisquamosus sp. n.; C. parvidentatus sp. n.; C. philippinensis sp. n.; C. simillimus sp. n.; and C. subaequalis sp. n. Lectotypes of following two valid species and three synonyms were designated: Curculio solani Fabricius, 1792; Curculio pulchellus Herbst, 1795; Cionus setiger Germar, 1821; Curculio immunis Marsham, 1802; and Cleopus pulchellus rigidus Stephens, 1831. Neotypes of Curculio perpensus Rossi, 1792 and Cleopus pulchellus flavus Stephens, 1832 were designated. The following new synonyms of Cleopus pulchellus (Herbst, 1795) were established: Cleopus pulchellus var. flavus Stephens, 1831 syn. n. and C. pulchellus var. rigidus Stephens, 1831 syn. n.

Morpho-Anatomy, In Vitro Culture, and Phylogenetic Studies of Two Helicotylenchus Species from Southern Alberta, Canada.

Spiral nematodes (Helicotylenchus spp.) are polyphagous parasitic species exhibiting a broad host range and geographical distribution. However, their diversity in the cultivated regions of southern Alberta remains understudied. Hence, we conducted a comprehensive survey of the region's arable lands for the presence of spiral nematodes and revealed two Helicotylenchus species, H. crassatus and H. oscephalus. H. crassatus consisted of two distinct morphotypes: one morphotype had a conoid tail with slight ventral projection on the distal end, whereas the other had a broadly rounded tail. This study presents the first documentation of H. crassatus and H. oscephalus from southern Alberta, Canada. Molecular characterization was based on the partial 18S rRNA, the D2-D3 of 28S rRNA, ITS rRNA, and COI gene sequences, complemented by detailed morphological studies using scanning electron microscopy. In this work, Helicotylenchus species were often co-detected with root lesion nematodes, which made the evaluation of their role in crop damage more difficult. To meet the requirements for threshold and pathogenicity assessments, we introduced both spiral nematode species to sterile carrot disks and evaluated the feasibility of their multiplication and mass production in vitro. The present findings expand the taxonomic records of Helicotylenchus spp. and improve diagnostics of these morphologically similar species. Furthermore, our in vitro culture technique will provide a reliable source of the initial inoculum for future plant-nematode interaction studies.

Optimizing malaria vector control in the Greater Mekong Subregion: a systematic review and mathematical modelling study to identify desirable intervention characteristics.

BACKGROUND: In the Greater Mekong Subregion (GMS), new vector-control tools are needed to target mosquitoes that bite outside during the daytime and night-time to advance malaria elimination. METHODS: We conducted systematic literature searches to generate a bionomic dataset of the main malaria vectors in the GMS, including human blood index (HBI), parity proportion, sac proportion (proportion with uncontracted ovary sacs, indicating the amount of time until they returned to host seeking after oviposition) and the resting period duration. We then performed global sensitivity analyses to assess the influence of bionomics and intervention characteristics on vectorial capacity. RESULTS: Our review showed that Anopheles minimus, An. sinensis, An. maculatus and An. sundaicus display opportunistic blood-feeding behaviour, while An. dirus is more anthropophilic. Multivariate regression analysis indicated that environmental, climatic and sampling factors influence the proportion of parous mosquitoes, and resting duration varies seasonally. Sensitivity analysis highlighted HBI and parity proportion as the most influential bionomic parameters, followed by resting duration. Killing before feeding is always a desirable characteristic across all settings in the GMS. Disarming is also a desirable characteristic in settings with a low HBI. Repelling is only an effective strategy in settings with a low HBI and low parity proportion. Killing after feeding is only a desirable characteristic if the HBI and parity proportions in the setting are high. CONCLUSIONS: Although in general adopting tools that kill before feeding would have the largest community-level effect on reducing outdoor transmission, other modes of action can be effective. Current tools in development which target outdoor biting mosquitoes should be implemented in different settings dependent on their characteristics.

Bionomics of malaria vectors in Lao PDR, 2018-2020: entomological surveillance as a key tool for malaria elimination.

BACKGROUND: The Lao PDR National Strategic Plan for malaria control and elimination for year 2021-2025 emphasizes the importance of routine entomological surveillance being conducted in areas with high transmission and in active malaria foci in elimination targeted areas. The collection of entomological surveillance data that is closely linked to recent epidemiological data is crucial for improving impact, as it contributes to the evidence package that supports operational and strategic decision-making of national malaria programmes, as they accelerate their last mile of elimination. METHODS: The Center for Malariology Parasitology and Epidemiology (CMPE) entomology team conducted entomological surveillance activities at 13 sentinel sites in 8 provinces and at active transmission foci sites from 2018 to 2020. The techniques used for the mosquito collection were indoor and outdoor human landing collections (from houses and from cultivation areas) and cattle baited net trap collections. RESULTS: There were 5601 Anopheles mosquito females captured and identified throughout the study, on both human and cow bait. They represented 15 different species or species complexes. The primary malaria vectors as well as the secondary vectors were present in all collection sites in the south, indicating that people living in these rural areas with high malaria incidence are exposed to the vectors. The vectors were highly zoophilic, but they still bite humans throughout the night with a high peak of activity before midnight, both indoors and outdoors. Overall, 17% of the malaria vectors were collected indoors when the people are sleeping. This confirms the importance of bed net use during the night. Thirty-two percent of primary and secondary vectors were collected outdoors at times when people are usually awake and outdoors, which shows that people are exposed to potentially infectious mosquitoes and the importance of personal protection at these times. The findings showed that residual transmission may occur outdoors in the villages, and outside the villages in cultivation fields and forested areas. Epidemiological data showed that transmission was higher in surveillance sites which were targeted as part of a malaria response rather than sentinel sites. CONCLUSIONS: Understanding where and how transmission is persisting, monitoring and mapping vector species distribution in areas with active transmission, monitoring biting trends, and designing evidence based and effective vector control interventions are critical to accelerating progress toward malaria elimination. In this context, the role of entomological surveillance combined with epidemiological data should be considered as a cornerstone in achieving malaria elimination.

Estimating malaria transmission risk through surveillance of human-vector interactions in northern Ghana.

BACKGROUND: Vector bionomics are important aspects of vector-borne disease control programs. Mosquito-biting risks are affected by environmental, mosquito behavior and human factors, which are important for assessing exposure risk and intervention impacts. This study estimated malaria transmission risk based on vector-human interactions in northern Ghana, where indoor residual spraying (IRS) and insecticide-treated nets (ITNs) have been deployed. METHODS: Indoor and outdoor human biting rates (HBRs) were measured using monthly human landing catches (HLCs) from June 2017 to April 2019. Mosquitoes collected were identified to species level, and Anopheles gambiae sensu lato (An. gambiae s.l.) samples were examined for parity and infectivity. The HBRs were adjusted using mosquito parity and human behavioral observations. RESULTS: Anopheles gambiae was the main vector species in the IRS (81%) and control (83%) communities. Indoor and outdoor HBRs were similar in both the IRS intervention (10.6 vs. 11.3 bites per person per night [b/p/n]; z = -0.33, P = 0.745) and control communities (18.8 vs. 16.4 b/p/n; z = 1.57, P = 0.115). The mean proportion of parous An. gambiae s.l. was lower in IRS communities (44.6%) than in control communities (71.7%). After adjusting for human behavior observations and parity, the combined effect of IRS and ITN utilization (IRS: 37.8%; control: 57.3%) on reducing malaria transmission risk was 58% in IRS + ITN communities and 27% in control communities with ITNs alone (z = -4.07, P < 0.001). However, this also revealed that about 41% and 31% of outdoor adjusted bites in IRS and control communities respectively, occurred before bed time (10:00 pm). The mean directly measured annual entomologic inoculation rates (EIRs) during the study were 6.1 infective bites per person per year (ib/p/yr) for IRS communities and 16.3 ib/p/yr for control communities. After considering vector survival and observed human behavior, the estimated EIR for IRS communities was 1.8 ib/p/yr, which represents about a 70% overestimation of risk compared to the directly measured EIR; for control communities, it was 13.6 ib/p/yr (16% overestimation). CONCLUSION: Indoor residual spraying significantly impacted entomological indicators of malaria transmission. The results of this study indicate that vector bionomics alone do not provide an accurate assessment of malaria transmission exposure risk. By accounting for human behavior parameters, we found that high coverage of ITNs alone had less impact on malaria transmission indices than combining ITNs with IRS, likely due to observed low net use. Reinforcing effective communication for behavioral change in net use and IRS could further reduce malaria transmission.

Composition of Anopheles species and bionomic characteristics over the peak malaria transmission season in Bandarban, Bangladesh.

BACKGROUND: Joint efforts by government and non-government organizations have helped to reduce malaria in Bangladesh and set the country on a clear path to eventual malaria elimination. However, achieving that goal would be challenging without a comprehensive understanding of vector bionomics. METHODS: Targeted capturing of Anopheles mosquitoes over a rainy season, utilizing specific sampling methods, including human landing catches (HLCs), CDC-light traps (CDC-LTs), and pyrethrum spray catches (PSCs) were aimed to characterize entomological drivers of transmission in four sites of Bandarban, Bangladesh. RESULTS: Molecular characterization of a subset of 4637 mosquitoes has demonstrated the presence of at least 17 species whose capture rates were representative of the rainy season. Species compositions and bionomic traits did not vary between sites with Anopheles maculatus having the highest landing rate by HLCs and Anopheles vagus having the highest capture rate with CDC-LTs. Interestingly, Anopheles species compositions and capture rates varied significantly (p < 0.05) for An. vagus, between HLCs and its often-used proxy-CDC-LTs- suggesting impacts on downstream analysis. CDC-LTs capture rates demonstrated differing compositions with indoor and outdoor biting rates. For example, Anopheles nigerrimus and Anopheles nivipes were more endophagic by HLCs and more exophagic by CDC-LTs. The use of a cow-baited CDC-LT also demonstrated significantly different results when compared to a human-baited CDC-LT considering the high degree of anthropophily in these species. The exception to both zoophily and indoor resting was An. vagus, which demonstrated both anthropophily and high resting rates indoors-pointing to this species being a possible primary vector at this site. CONCLUSION: A diverse Anopheles fauna in Bandarban has been confirmed through molecular methods, highlighting the potential impact of sampling techniques. Given the complexity of the local ecosystem, a better understanding of mosquito behaviour and ecology is required to achieve the goal of malaria elimination in Bangladesh.

Comparative bionomics and life table studies of Lipaphis erysimi pseudobrassicae (Davis) and Myzus persicae (Sulzer) (Hemiptera: Aphididae) on three cabbage varieties.

Lipaphis erysimi pseudobrassicae (Davis) and Myzus persicae (Sulzer) are important pests of brassica crops, causing significant yield losses on cabbage in Ghana. To inform the development of ecologically sound and sustainable pest management strategies for these pests, their biological and population growth parameters were studied on three cabbage varieties (Oxylus, Fortune, and Leadercross). The study was conducted in a screen house under ambient conditions at 30 ± 1°C and 75 ± 5% RH and 12:12 h photoperiod from September to November 2020. The parameters of the preadult developmental period, survival rates, longevity, reproduction, and life table were evaluated following the female age-specific life table. There were significant differences in the nymphal developmental time, longevity, and fecundity on the cabbage varieties for both aphid species. The highest population growth parameters, net reproductive rate (R0), intrinsic rate of increase r, and finite rate of increase (λ) were recorded on Oxylus variety for both L. e. pseudobrassicae and M. persicae. The lowest was recorded on Leadercross variety for L.e pseudobrassicae and Fortune for M. persicae. The results from this study suggest that Leadercross is a less suitable host for L. e. pseudobrassicae and Fortune for M. persicae, thus, should be considered as less susceptible varieties for use in primary pest management by small-scale farmers or as a component of an integrated pest management strategy for these pests on cabbage.

Entomological monitoring data driving decision-making for appropriate and sustainable malaria vector control in Côte d'Ivoire.

BACKGROUND: Entomological surveillance provides critical information on vectors for appropriate malaria vector control and strategic decision-making. The widely documented insecticide resistance of malaria vectors in Côte d'Ivoire requires that any vector control intervention deployment be driven by entomological data to optimize its effectiveness and appropriate resource allocations. To achieve this goal, this study documents the results of monthly vector surveillance and insecticide susceptibility tests conducted in 2019 and a review of all previous entomological monitoring data used to guide vector control decision making. Furthermore, susceptibility to pirimiphos-methyl and clothianidin was assessed in addition to chlorfenapyr and pyrethroids (intensity and piperonyl butoxide (PBO) synergism) tests previously reported. Vector bionomic data were conducted monthly in four sites (Sakassou, Béoumi, Dabakala and Nassian) that were selected based on their reported high malaria incidence. Adult mosquitoes were collected using human landing catches (HLCs), pyrethrum spray catches (PSCs), and human-baited CDC light traps to assess vector density, behaviour, species composition and sporozoite infectivity. RESULTS: Pirimiphos-methyl and clothianidin susceptibility was observed in 8 and 10 sites, respectively, while previous data reported chlorfenapyr (200 µg/bottle) susceptibility in 13 of the sites, high pyrethroid resistance intensity and increased mortality with PBO pre-exposure at all 17 tested sites. Anopheles gambiae sensu lato was the predominant malaria vector collected in all four bionomic sites. Vector density was relatively higher in Sakassou throughout the year with mean biting rates of 278.2 bites per person per night (b/p/n) compared to Béoumi, Dabakala and Nassian (mean of 48.5, 81.4 and 26.6 b/p/n, respectively). The mean entomological inoculation rate (EIR) was 4.44 infective bites per person per night (ib/p/n) in Sakassou, 0.34 ib/p/n in Beoumi, 1.17 ib/p/n in Dabakala and 1.02 ib/p/n in Nassian. The highest EIRs were recorded in October in Béoumi (1.71 ib/p/n) and Nassian (3.22 ib/p/n), in July in Dabakala (4.46 ib/p/n) and in May in Sakassou (15.6 ib/p/n). CONCLUSION: Based on all results and data review, the National Malaria Control Programme developed and implemented a stratified insecticide-treated net (ITN) mass distribution in 2021 considering new generation ITNs. These results also supported the selection of clothianidin-based products and an optimal spraying time for the first indoor residual spraying (IRS) campaign in Sakassou and Nassian in 2020.

Entomological Surveillance Planning Tool (ESPT)-generated actionable evidence on human and vector behaviours optimizes present interventions and reduces exposure to Anopheles vectors in two communities of Guna Yala, Panamá.

BACKGROUND: Although most of Panamá is free from malaria, localized foci of transmission persist, including in the Guna Yala region. Government-led entomological surveillance using an Entomological Surveillance Planning Tool (ESPT) sought to answer programmatically relevant questions on local entomological drivers of transmission and gaps in protection to guide local vector control decision-making. METHODS: The ESPT was used to design a sampling plan to answer priority programmatic questions about the appropriateness of Long Lasting Insecticidal Nets (LLINs) and spaces and times where humans remain exposed to Anopheles bites (gaps in protection) in the communities of Permé and Puerto Obaldía, Guna Yala. Adult Anopheles were sampled at three time points via human landing catches (HLCs) during the rainy and dry seasons (2018/2019). Human behaviour observations (HBOs) were conducted alongside HLCs to examine intervention use, indoor versus outdoor activity, and sleeping patterns. HLC and HBO data were integrated to evaluate HBO-adjusted human biting rate (HBR). RESULTS: A total of 7,431 adult Anopheles were collected across both sites. Of the 450 specimens molecularly confirmed to species-level, 75.5% (n = 340) were confirmed as Anopheles Nyssorhynchus albimanus, followed by Anopheles (Ny.) aquasalis. Anopheles host seeking activity was demonstrated to be primarily exophagic throughout all sampling periods and in both communities. When adjusted with HBOs, exposure to mosquito bites was predominantly indoors and overnight in Permé (Nov, Mar), compared to predominantly outdoors in Puerto Obaldía (Nov, Mar, Jul). Differences in site-specific human-vector exposure profiles were due to contrasting cultural and lifestyle practices between Permé and Puerto Obaldía (possibly partly influenced by the absence of electricity in Permé), and lower LLIN use in Permé. This evidence supported a previously planned LLIN campaign alongside a social behaviour change communication (SBCC) strategy in the Guna Yala Comarca (Jul 2019), which increased LLIN use. In turn, this led to a reduction of indoor exposure to mosquito bites, and a shift to predominant outdoor exposure to mosquito bites. CONCLUSION: ESPT-based question-driven planning and the integration of HBOs, intervention, and HLC data generated evidence towards answering the programmatic questions. This evidence enabled the characterization of site-specific human-vector exposure profiles, and the quantification of remaining gaps in protection. These data also provide important insights into remaining gaps in protection that must be addressed to further reduce human exposure to mosquito bites at these sites.

Biology, bionomics and life-table studies of Anopheles stephensi (Diptera: Culicidae) in Sri Lanka and estimating the vectorial potential using mathematical approximations.

BACKGROUND: Anopheles stephensi is an invasive mosquito in Sri Lanka that can potentially transmit malaria. The transmission intensity is linked with biology, bionomic and behavioral aspects of a vector that are associated with the Vectorial Capacity (VC). However, the influence of larval conditions eventually affects the vectorial potential of An. stephensi are not well understood. METHODS: A colony of An. stephensi was established at the Regional Centre of the Open University of Sri Lanka, Jaffna District. The colony was maintained under confined conditions according to standard protocols. Biotypes of An. stephensi were characterized by referring to the number of egg ridges. Information on (a) biological aspects of eggs (duration for egg hatching, egg development and hatchability), (b) larval development time, larval survivorship pupation success, resting depth of larvae), (c) pupae (adult emergence rate, average time for adult emergence) and (d) adults (biting frequency, mating success gonotrophic cycle, fecundity, duration for egg-laying, percentage of sexes, adult survival/longevity) were evaluated under life-table analysis. Further, selected morphometric characters of each life cycle stage were recorded from the eggs (length and breadth), larvae (head length, width of head, length of thorax, width of thorax, length of abdomen, width of abdomen, and the total length of larvae), pupae (cephalothoracic length and width) and adults (length & width of wing, thorax and abdomen). The VC was calculated using a mathematical-based approach. Descriptive statistics, General Linear Model (GLM) and independent-sample t-test were used for the statistical analysis. RESULTS: All three biotypes were identified based on egg morphology. Mysorensis biotype (47%; n = 470) was predominant followed by type (38.1%; n = 381) and intermediate (14.9%; n = 149). The mean egg length (F(2,997) = 3.56; P = 0.029) and breadth (F(2,997) = 4.57; P = 0.011) denoted significant differences among the three biotypes. The mating success of females observed was 80.7 ± 4.45%. The mean hatching period was 1.9 ± 0.03 days, with a hatching rate of 86.2 ± 0.77%. Overall, 8.0 ± 0.14 days were required for larval development and 30.3 ± 0.14 h were spent in the pupal stage. The pupation success was 94.5 ± 0.37%, and the majority were males (53.1 ± 0.73%). The mean fecundity was 106.5 ± 6.38 eggs and a gonotrophic cycle of 3.4 ± 0.06 days. The female survival rate was 43.2 ± 2.4%, with a mean biting frequency of 66.6 ± 3.5%. The average VC of adult An. stephensi was estimated to be 18.7. CONCLUSIONS: The type biotype, which is an effective vector in the Indian subcontinent is present in Sri Lanka. According to the mathematical approximation, An. stephensi found locally has a vectorial capacity of over 18. Therefore, this study warrants the health authorities and vector control programmes to continue the entomological surveys, monitoring of vector densities and implementing appropriate vector control interventions based on biology and bionomic information of vectors.

Malaria vector bionomics and transmission in irrigated and non-irrigated sites in western Kenya.

Irrigation not only helps to improve food security but also creates numerous water bodies for mosquito production. This study assessed the effect of irrigation on malaria vector bionomics and transmission in a semi-arid site with ongoing malaria vector control program. The effectiveness of CDC light traps in the surveillance of malaria vectors was also evaluated relative to the human landing catches (HLCs) method. Adult mosquitoes were sampled in two study sites representing irrigated and non-irrigated agroecosystems in western Kenya using a variety of trapping methods. The mosquito samples were identified to species and assayed for host blood meal source and Plasmodium spp. sporozoite infection using polymerase chain reaction. Anopheles arabiensis was the dominant malaria vector in the two study sites and occurred in significantly higher densities in irrigated study site compared to the non-irrigated study site. The difference in indoor resting density of An. arabiensis during the dry and wet seasons was not significant. Other species, including An. funestus, An. coustani, and An. pharoensis, were collected. The An. funestus indoor resting density was 0.23 in irrigated study site while almost none of this species was collected in the non-irrigated study site. The human blood index (HBI) for An. arabiensis in the irrigated study site was 3.44% and significantly higher than 0.00% for the non-irrigated study site. In the irrigated study site, the HBI of An. arabiensis was 3.90% and 5.20% indoor and outdoor, respectively. The HBI of An. funestus was 49.43% and significantly higher compared to 3.44% for An. arabiensis in the irrigated study site. The annual entomologic inoculation rate for An. arabiensis in the irrigated study site was 0.41 and 0.30 infective bites/person/year indoor and outdoor, respectively, whereas no transmission was observed in the non-irrigated study site. The CDC light trap performed consistently with HLC in terms of vector density. These findings demonstrate that irrigated agriculture may increase the risk of malaria transmission in irrigated areas compared to the non-irrigated areas and highlight the need to complement the existing malaria vector interventions with novel tools targeting the larvae and both indoor and outdoor biting vector populations.

Diapause characterization in the invasive alien mosquito species Aedes koreicus: a laboratory experiment.

Aedes koreicus is an invasive alien mosquito species native to Asia now introduced in several European countries, including northern Italy. In this temperate region, mosquito populations survive cold winter temperatures thanks to diapausing eggs or adults, depending on the species. In its native area, Ae. koreicus was reported to overwinter in the egg stage, but to the best of our knowledge, it is not confirmed whether overwintering eggs are actually diapausing or only in a quiescence stage, i.e., they might hatch as soon as external conditions are favorable. Based on previous laboratory studies, we established a diapausing Ae. koreicus colony, maintained at 21 °C with a photoperiod of 12L:12D. Females were allowed to lay eggs, which were consequently placed in water at different time intervals after oviposition, from 30 days to 5 months. We found that diapausing eggs younger than 3 months have a poor hatching rate, while after about 100 days we observed that almost all eggs hatched. Our findings highlight that water immersion alone did not lead to the hatching of eggs, as age was found to be a significantly important factor. We thus confirm effective diapause, occurring at the egg stage, for Ae. koreicus in a recently invaded area. Moreover, our quantification of diapause duration and hatching success might help in better designing future experiments and improving modeling efforts.

Morphological and odorant-binding protein 1 gene intron 1 sequence variations in Anopheles stephensi from Jaffna city in northern Sri Lanka.

Three Anopheles stephensi biotypes have historically been differentiated through variations in the mode numbers of egg ridges and adult spiracular indices. Anopheles stephensi odorant-binding protein 1 gene (AsteObp1) sequences in Iran and Afghanistan have been recently interpreted to suggest that the three biotypes are sibling species. AsteObp1 intron 1 sequences, mode numbers of egg ridges and spiracular indices of An. stephensi in Jaffna city in Sri Lanka were therefore investigated in field-collected mosquitoes and short-term laboratory colonies established from them. AsteObp1 intron 1 sequences revealed the region to be polymorphic with four unique sequences, ASJF1-4, present in both short-term laboratory colonies and field-collected An. stephensi. The spiracular index did not relate to the mode number of egg ridges in Jaffna An. stephensi. The results suggested that numbers of egg ridges, spiracular indices and AsteObp1 intron 1 sequences were not useful for differentiating An. stephensi biotypes in Jaffna. It is proposed that the observed differences between An. stephensi mosquitoes in Jaffna now result from normal population variance in the context of rapidly changing bionomics in India and northern Sri Lanka.

Life-history attributes of juvenile Anopheles gambiae s.s. in central Uganda; implications for malaria control interventions.

Malaria is among the leading causes of death in Uganda, and Anopheles gambiae sensu stricto (s.s.) is the predominant vector. Although current vector control interventions have greatly reduced the malaria burden, the disease persists. New interventions are needed in order to eradicate them. Evaluation of new tools will require the availability of well-characterized test vector populations. Juvenile An. gambiae s.s. from Kibbuye and Kayonjo-derived populations were characterized under semi-field and laboratory conditions, given that various vector traits, including abundance and fitness are dependent on development profiles at this life stage. Ten replicates comprising 30 first instar larvae each were profiled for various life-history attributes (egg hatching, larval development time, larval survivorship, pupal weight and pupation rate). All parameters were similar for the two sites under laboratory conditions. However, the similarities or differences between field and laboratory development were parameter-specific. Whereas, larval survivorship and pupal weight were similar across seasons and laboratory in colonies from both sites, in the semi-field settings, pupation rate and larval survivorship differed between seasons in both sites. In addition, the average larval development time during the wet season was longer than that of the laboratory for both sites. Availability of mirror field sites is important for future tool evaluations.

Life cycle and description of the immature stages of a terrestrial firefly endemic to Mexico: Photinusextensus Gorham (Coleoptera, Lampyridae).

The life cycle, morphology, and bionomy of Photinusextensus Gorham, 1881, an endemic species of Mexico, are described. Redescriptions of adults (male and female) are also presented. Larvae were reared to the adult stage from eggs laid by females collected at the El Pedregal de San Ángel Ecological Reserve, south of Mexico City. The activity period of adults of P.extensus begins at the end of July and finishes by the end of August. Females lay between 3 and 198 eggs. Larvae hatch from the eggs after a period of 11 to 71 days, undergo 6 larval instars and a pupal stage in an annual cycle. Morphological characters of the sixth larval instar of P.extensus are compared with those of several other genera assigned to the tribe Photinini. Knowledge of the natural history of firefly larvae is relevant since most species do not feed as adults and therefore depend on resources acquired during the larval stage.

Anopheles drivers of persisting malaria transmission in Guna Yala, Panamá: an operational investigation.

BACKGROUND: Though most of Panamá is free from malaria, localized foci of transmission persist, including in the Guna Yala region. Government-led entomological surveillance using an entomological surveillance planning tool (ESPT) sought to answer programmatically-relevant questions that would enhance the understanding of both local entomological drivers of transmission and gaps in protection that result in persisting malaria transmission to guide local vector control decision-making. METHODS: The ESPT was used to design a sampling plan centered around the collection of minimum essential indicators to investigate the relevance of LLINs and IRS in the communities of Permé and Puerto Obaldía, Guna Yala, as well as to pinpoint any remaining spaces and times where humans are exposed to Anopheles bites (gaps in protection). Adult Anopheles were collected at three time points via human landing catches (HLCs), CDC Light Traps (LT), and pyrethrum spray catches (PSCs) during the rainy and dry seasons. Mosquitoes were identified to species via molecular methods. Insecticide susceptibility testing of the main vector species to fenitrothion was conducted. RESULTS: In total, 7537 adult Anopheles were collected from both sites. Of the 493 specimens molecularly confirmed to species, two thirds (n = 340) were identified as Nyssorhynchus albimanus, followed by Anopheles aquasalis. Overall Anopheles human biting rates (HBRs) were higher outdoors than indoors, and were higher in Permé than in Puerto Obaldía: nightly outdoor HBR ranged from 2.71 bites per person per night (bpn) (Puerto Obaldía), to 221.00 bpn (Permé), whereas indoor nightly HBR ranged from 0.70 bpn (Puerto Obaldía) to 81.90 bpn (Permé). Generally, peak biting occurred during the early evening. The CDC LT trap yields were significantly lower than that of HLCs and this collection method was dropped after the first collection. Pyrethrum spray catches resulted in only three indoor resting Anopheles collected. Insecticide resistance (IR) of Ny. albimanus to fenitrothion was confirmed, with only 65.5% mortality at the diagnostic time. CONCLUSION: The early evening exophagic behaviour of Anopheles vectors, the absence of indoor resting behaviours, and the presence of resistance to the primary intervention insecticide demonstrate limitations of the current malaria strategy, including indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs), and point to both gaps in protection and to the drivers of persisting malaria transmission in Guna Yala. These findings highlight the need for continued and directed entomological surveillance, based on programmatic questions, that generates entomological evidence to inform an adaptive malaria elimination strategy.

Culex nigripalpus Distribution Expansion: First Record in Virginia, New County Records in North Carolina, and Revised United States Map.

Theobald first described Culex nigripalpus in 1901 from St. Lucia Island of the Lesser Antilles. It is a Neotropical mosquito species that is documented throughout Central and South America, the Caribbean, and the southeastern USA. Reports within the last 3 decades show the species has expanded its range farther into North America and is not only being discovered sporadically but appears to be establishing itself in several states, of which data are underreported and difficult to find. Five female specimens were collected-4 on October 13 and 1 on December 6, 2017-at 3 separate sites in the city of Suffolk, VA-2 in a BG-Sentinel 2® trap, 3 in Centers for Disease Control and Prevention light traps-during routine mosquito surveillance; subsequent collections were made in 2019 and 2020. These findings represent the 1st record of Cx. nigripalpus in the state of Virginia and the most northeastern records in the USA. An updated mosquito species checklist for the state of Virginia is provided. Additionally, this report serves to update the records of North Carolina to 28 total counties reporting Cx. nigripalpus. These new records, along with other reports and published studies, show this species now documented in 201 jurisdictions in 16 states of the USA. As recent literature on this species is scarce, we provide a review of the bionomics, defining morphological characters, and an updated US distribution map.

The vectors of Plasmodium knowlesi and other simian malarias Southeast Asia: challenges in malaria elimination.

Plasmodium knowlesi, a simian malaria parasite of great public health concern has been reported from most countries in Southeast Asia and exported to various countries around the world. Currently P. knowlesi is the predominant species infecting humans in Malaysia. Besides this species, other simian malaria parasites such as P. cynomolgi and P. inui are also infecting humans in the region. The vectors of P. knowlesi and other Asian simian malarias belong to the Leucosphyrus Group of Anopheles mosquitoes which are generally forest dwelling species. Continual deforestation has resulted in these species moving into forest fringes, farms, plantations and human settlements along with their macaque hosts. Limited studies have shown that mosquito vectors are attracted to both humans and macaque hosts, preferring to bite outdoors and in the early part of the night. We here review the current status of simian malaria vectors and their parasites, knowledge of vector competence from experimental infections and discuss possible vector control measures. The challenges encountered in simian malaria elimination are also discussed. We highlight key knowledge gaps on vector distribution and ecology that may impede effective control strategies.