Anopheline Species Research Articles

Identification of a biological form in the Anopheles stephensi laboratory colony using the odorant-binding protein 1 intron I sequence.

Anopheles stephensi Listen (1901) is a major vector of malaria in Asia and has recently been found in some regions of Africa. The An. stepehnsi species complex is suspected to have three sibling species: type, intermediate, and mysorensis, each with its own vector competence to the malaria parasite and ecology. To identify the members of the species complex in our An. stephensi insectary colony, we used the morphological features of eggs and genetic markers such as AnsteObp1 (Anopheles stephensi odorant binding protein 1), mitochondrial oxidases subunit 1 and 2 (COI and COII), and nuclear internal transcribed spacer 2 locus (ITS2). Eggs were collected from individual mosquitoes (n = 50) and counted for the number of ridges under stereomicroscope. Genomic DNA was extracted from female mosquitoes. After the amplification of partial fragments of AnsteObp1, COI, COII and ITS2 genes, the PCR products were purified and sequenced. Phylogenetic analysis was performed after aligning query sequences against the submitted sequences in GenBank using MEGA 7. The range of ridges number on each egg float was 12-13 that corresponds to the mysorensis form of An. stephensi. The generated COI, COII and ITS2 sequences showed 100%, 99.46% and 99.29% similarity with the sequences deposited for Chinese, Indian and Iranian strains of An. stephensi, respectively. All the generated AnsteObp1 intron I region sequences matched 100% with the sequences deposited for An. stephensi sibling species C (mysorensis form) from Iran and Afghanistan. This manuscript precisely describes the morphological and molecular details of the 'var mysorensis' form of An. stephensi that could be exploited in elucidating its classification as well as in differentiation from other biotypes of the same or other anopheline species. Based on our findings, we recommend AnsteObp1 as a robust genetic marker for rapid and accurate discrimination (taxonomic identification) of the An. stephensi species complex, rather than the COI, COII, and ITS2 marker, which could only be utilized for interspecies (Anopheles) differentiation.

Discrimination of 15 Amazonian Anopheline Mosquito Species by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism.

Precise identification of anopheline species is paramount for incrimination of malaria vectors and implementation of a sustainable control program. Anopheline mosquitoes are routinely identified morphologically, a technique that is time-consuming, needs high level of expertise, and prone to misidentifications especially when considering Amazonian species. The aim of this study was therefore to develop a DNA-based identification technique to supplement traditional morphological identification methods for the discrimination of anopheline mosquitoes collected in French Guiana. The internal transcribed spacer 2 (ITS2) region of ribosomal DNA (rDNA) for anopheline species was amplified by polymerase chain reaction (PCR), and digested with AluI/MspI restriction enzymes. PCR-restriction fragments length polymorphism (RFLP) assay was compared to sequencing of the ITS2 region for validation. Fifteen Anopheles species have shown distinct PCR-RFLP profiles. A concordance of 100% was obtained when identification by PCR-RFLP was compared to sequencing of ITS2. A high throughput, fast, and cost-effective PCR-RFLP assay has been developed for unambiguous discrimination of fifteen anopheline mosquito species from French Guiana including primary and suspected secondary malaria vectors.

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.

Characterization of the anopheline vector breeding habitats: Implications for elimination of malaria in tribal inhabited Car Nicobar Island.

Malaria is an important public health problem in Andaman & Nicobar archipelago. Among the three districts, Nicobar is the most endemic district where API is >2. In this district, the malaria incidence in Car Nicobar Tehsil has declined steadily over the past 10 years. A renewed initiative to consolidate this gain is being made with the ultimate objective of achieving zero indigenous transmission of malaria in Car Nicobar. So, the present study undertook a close environmental monitoring of water bodies for assessing changes in the risk potential of mosquito vector breeding habitats which can augment the elimination programme. The breeding habitats of anopheline mosquitoes were sampled in 16 areas of Car Nicobar Island for eight time periods during 2017-2020. Along with anophelines, various associated water parameters (n=60) were estimated, viz. physicochemical (n=13), and biological, which included culicine mosquito immatures, insect predators (n=5), phytoplanktons (n=31) and zooplanktons (n=10). In the 16 study sites, overall 1126 surface water stagnating bodies constituting 21 different habitat types were surveyed. Of these, 17 were positive for anopheline breeding. Water bodies from three villages were consistently found to be positive for anopheline breeding. However, early instars of anopheline larvae were more abundant compared to the late instars. Four anopheline species were recorded, including Anopheles sundaicus, A. barbirostris, A. insulaeflorum and A. subpictus, in which 48 per cent were A. sundaicus. Multivariable analysis indicated that anopheline density was significantly higher in permanent water bodies than in temporary habitats (P<0.05) (high risk of anophelines). The highest pH (≥8.2), dissolved solids (≥0.39) levels showed significantly (P<0.05) decreased larval densities (lower risk of breeding), adjusted with breeding sites and season. Nitrite levels increased (P=0.022) larval densities. The present study facilitated estimating the productive period of a larval habitat enabling target larval sources to reduce adult populations. Implementing larviciding strategy before monsoon season is presumably the most cost-effective strategy. The output can be utilized for environmental monitoring of mosquito breeding risk in other malaria endemic areas, particularly where medium/large water bodies are the predominant breeding sites for malaria vectors.

Phylogenetic relationships of the supercontig of sodium channel subunit I (NaV) in 17 species of Anopheles (Diptera: Culicidae).

Malaria is a global health problem and is transmitted by the Anopheles species. Due to the epidemiological importance of the genus, studies on biological, phylogenetic, and evolutionary aspects have contributed to the understanding of adaptation, vector capacity, and resistance to insecticides. The latter may result from different causes such as mutations in the gene that encodes the sodium channel (NaV). In this study, the NaV subunit I scaffold of 17 anopheline species was used to infer phylogenetic relationships of the genus Anopheles using Bayesian inference. The evolutionary phylogenetic tree of the NaV gene was aligned in the AliView program and analyzed utilizing Bayesian inference, using the software MrBayes. The anophelines were grouped into five well-supported clusters: 1 - Anopheles darlingi and Anopheles albimanus; 2 - Anopheles sinensis and Anopheles atroparvus; 3 - Anopheles dirus; 4 - Anopheles minimus, Anopheles culicifacies, Anopheles funestus, Anopheles maculatus, and Anopheles stephensi; and 5 - Anopheles christyi, Anopheles epiroticus, Anopheles merus, Anopheles melas, Anopheles gambiae, Anopheles coluzzii, and Anopheles arabiensis. The topology confirms the phylogenetic relationships proposed in studies based on the genome of some anophelines and reflects the current taxonomy of the genus, which suggests that NaV undergoes selection pressure during the evolution of the species. These data are useful tools for inferring their ability to resist insecticides and also help in better understanding the evolutionary processes of the genus Anopheles.

Functional conservation of Anopheline linalool receptors through 100 million years of evolution.

Insects rely on olfactory receptors to detect and respond to diverse environmental chemical cues. Detection of semiochemicals by these receptors modulates insect behavior and has a direct impact on species fitness. Volatile organic compounds (VOCs) are released by animals and plants and can provide contextual cues that a blood meal host or nectar source is present. One such VOC is linalool, an enantiomeric monoterpene, that is emitted from plants and bacteria species. This compound exists in nature as one of two possible stereoisomers, (R)-(-)-linalool or (S)-(+)-linalool. In this study, we use a heterologous expression system to demonstrate differential responsiveness of a pair of Anopheline odorant receptors (Ors) to enantiomers of linalool. The mosquitoes Anopheles gambiae and Anopheles stephensi encode single copies of Or29 and Or53, which are expressed in the labella of An. gambiae. (S)-(+)-linalool activates Or29 orthologs with a higher potency than (R)-(-)-linalool, while the converse is observed for Or53 orthologs. The conservation of these receptors across a broad range of Anopheline species suggests they may function in the discrimination of linalool stereoisomers, thereby influencing the chemical ecology of mosquitoes. One potential application of this knowledge would be in the design of novel attractants or repellents to be used in integrated pest management practices.

Anopheline species (Diptera: Culicidae) from the Maracá Ecological Station, Roraima State, Brazil

Anopheline species were sampled in different ecotones at Maracá Ecological Station and in Cantá municipality, both in Roraima State, Brazil, in June 2016. Nine species were recorded. Six species belong to the subgenus Nyssorhynchus and three species to the subgenus Anopheles. The most abundant species collected were Anopheles triannulatus s.l., Anopheles nuneztovari s.l. and Anopheles oswaldoi species B. The remaining species had frequencies lower than 10%. While A. triannulatus s.l. was the most frequent and the most widely distributed species in the studied area, Anopheles darlingi Root, the main malaria vector in the Brazilian Amazon region, was captured in two different ecotones and in low frequency. Our findings suggest that the risk of malaria outbreaks at Maracá Ecological Station may be low. This study also describes the first record of Anopheles costai Fonseca &amp; Ramos in Maracá Ecological Station, Roraima State, increasing the distribution range of this taxon.

Phylogeny of Anopheles darlingi (Diptera:Culicidae) based on the antimicrobial peptide genes cecropin and defensin

Cecropins and defensins are the main classes of antimicrobial peptides in the mosquito innate immune system, acting against bacteria, fungi and protozoa. There is a knowledge gap concerning these peptide genes in anopheline mosquitoes from the Brazilian Amazon. Thus, this work aimed to describe molecular techniques for detecting the genes encoding the antimicrobial peptides cecropin A (CecA) and defensin in Anopheles darlingi mosquitoes and to perform molecular phylogeny of the sequenced genes using the maximum likelihood method and Bayesian inference with other species from different geographic areas. Our results show, for the first time, a molecular biology method for detecting CecA and defensin in Anopheles darlingi that allows for the use of these molecular markers for phylogenetic analysis in anopheline species, separating the species into single and monophyletic clades.

Molecular Analysis Reveals a High Diversity of Anopheline Mosquitoes in Yanomami Lands and the Pantanal Region of Brazil.

Identifying the species of the subfamily Anophelinae that are Plasmodium vectors is important to vector and malaria control. Despite the increase in cases, vector mosquitoes remain poorly known in Brazilian indigenous communities. This study explores Anophelinae mosquito diversity in the following areas: (1) a Yanomami reserve in the northwestern Amazon Brazil biome and (2) the Pantanal biome in southwestern Brazil. This is carried out by analyzing cytochrome c oxidase (COI) gene data using Refined Single Linkage (RESL), Assemble Species by Automatic Partitioning (ASAP), and tree-based multi-rate Poisson tree processes (mPTP) as species delimitation approaches. A total of 216 specimens collected from the Yanomami and Pantanal regions were sequenced and combined with 547 reference sequences for species delimitation analyses. The mPTP analysis for all sequences resulted in the delimitation of 45 species groups, while the ASAP analysis provided the partition of 48 groups. RESL analysis resulted in 63 operational taxonomic units (OTUs). This study expands our scant knowledge of anopheline species in the Yanomami and Pantanal regions. At least 18 species of Anophelinae mosquitoes were found in these study areas. Additional studies are now required to determine the species that transmit Plasmodium spp. in these regions.

Anopheline Diversity in Indonesia: An Evaluation of Animal-Baited Sampling Techniques.

Indonesia has rich Anopheline (Diptera: Culicidae) mosquito species living in various types of ecosystems. The study was conducted to profile and compare Anopheles diversity, equitability, and dominance in various ecosystems using different animal-based sampling techniques. The present study analyzed a subset of data collected from a nation-wide vector and animal reservoirs survey in 2016. Analyses were restricted to three ecosystem types (forest, nonforest, and coastal areas) in Java and Sumatera Islands. A total of 5,477 Anopheles were collected by using animal-baited (n = 1,909) and animal-baited trap nets (n = 1,978), consisting of 23 Anopheline species. Overall, Anopheles vagus was the most abundant species, followed by An. subpictus and An. barbirostris. Among the three ecosystems, the forest had a higher diversity index (H' = 1.98), but each ecosystem has its specific predominant species. Compared with the animal-baited method, the Anopheles abundance collected by animal-baited trap nets was two-fold higher. Ecosystem, elevation, and sampling methods were associated with the abundance of female Anopheles (P-value < 0.001). Our findings revealed that Anopheles were found in a different ecosystem, indicating the potential of malaria transmission. This suggests that improved malaria vector surveillance is essential in all types of ecosystem. Furthermore, the study suggested that animal-baited trap nets could be used as the standard method of outdoor resting sampling in Indonesia in addition to the traditional human landing collection approach.

Entomological Surveillance in Former Malaria-endemic Areas of Southern Italy.

Malaria still represents a potential public health issue in Italy, and the presence of former Anopheles vectors and cases imported annually merit continuous surveillance. In areas no longer endemic, the concurrent presence of gametocyte carriers and competent vectors makes re-emergence of local transmission possible, as recently reported in Greece. In October 2017, due to the occurrence of four suspected introduced malaria cases in the province of Taranto (Apulia region), entomological investigations were performed to verify the involvement of local anopheline species. In 2019–2020 entomological surveys were extended to other areas historically prone to malaria between the provinces of Taranto and Matera and the province of Foggia (Gargano Promontory). Resting mosquitoes were collected in animal shelters and human dwellings, larvae were sampled in natural and artificial breeding sites, and specimens were both morphologically and molecularly identified. A total of 2228 mosquitoes were collected, 54.3% of which were anophelines. In all the investigated areas, Anopheles labranchiae was the most widespread species, while Anopheles algeriensis was predominant at the Gargano sites, and Anopheles superpictus and Anopheles plumbeus were recorded in the province of Matera. Our findings showed a potentially high receptivity in the surveyed areas, where the abundance of the two former malaria vectors, An. labranchiae and An. superpictus, is related to environmental and climatic parameters and to anthropic activities.

The effect of irrigation on malaria vector bionomics and transmission intensity in western Ethiopia

BackgroundIrrigation schemes may result in subsequent changes in malaria disease dynamics. Understanding the mechanisms and effects of irrigation on malaria vector bionomics and transmission intensity is essential to develop new or alternative surveillance and control strategies to reduce or control malaria risk. This study was designed to assess the effect of rice irrigation on malaria vector bionomics and transmission intensity in the Gambella Region, Ethiopia.MethodsComparative cross-sectional study was conducted in Abobo District of the Gambella Region, Ethiopia. Accordingly, clusters (kebeles) were classified into nearby and faraway clusters depending on their proximity to the irrigation scheme. Adult mosquito survey was conducted in February, August and November 2018 from three nearby and three faraway clusters using Centers for Disease Control and Prevention (CDC) light traps (LTs). During the November survey, human landing catch (HLC) and pyrethrum spray catch (PSC) were also conducted. The collected mosquitoes were morphologically identified to species and tested for Plasmodium infection using circumsporozoite protein enzyme-linked immunosorbent assay (CSP-ELISA). Furthermore, species-specific polymerase chain reaction (PCR) was performed to identify member species of the Anopheles gambiae complex. Chi-square and t-tests were used to analyze the data using the SPSS version 20 software package.ResultsA total of 4319 female anopheline mosquitoes comprising An. gambiae sensu lato, An. funestus group, An. pharoensis, An. coustani complex and An. squamosus were collected. Overall, 84.5% and 15.5% of the anopheline mosquitoes were collected from the nearby and faraway clusters, respectively. Anopheles gambiae s.l. was the predominant (56.2%) anopheline species in the area followed by An. pharoensis (15.7%). The density of anopheline mosquitoes was significantly higher in the nearby clusters in both HLCs [t(3) = 5.14, P = 0.0143] and CDC LT catches [t(271.97) = 7.446, P < 0.0001). The overall sporozoite rate of anopheline species from the nearby clusters was 10-fold higher compared to the faraway clusters.ConclusionsSignificantly higher mosquito population density was observed in areas close to the irrigation sites. Sporozoite infection rate in the mosquito population was also markedly higher from the nearby clusters. Therefore, the irrigation scheme could increase the risk of malaria in the area.Graphical abstract

Mobile app based pictorial identification key for Indian anophelines

Malaria control strategies for a particular area largely depends on the correct identification of the mosquito species prevalent in that area. This study aimed to develop an android mobile app named 'Pictorial Indian anophelines key', which can be used to identify the 58 species of Indian female adult anophelines based on morphological characteristics. The pre-requisite to use this app requires a minimum basic knowledge of mosquito morphological characteristics and features. The user needs to view the collected mosquito under a dissecting binocular microscope and has to select the observations on the app presented in the form of labelled picture format. The app uses a divide and conquer algorithm which narrows the selection from genus level to final identification of species. The app is user-friendly, quick and it accurately identifies the mosquito species in a maximum of six consecutive clicks. The result displays a complete image of identified mosquito along with its quick identification features. It can be used for both subgenus Cellia and Anopheles. This pictorial key-based mobile app could be helpful for entomologists, malariologists, researchers, public health workers as well as students in the quick and correct identification of anopheline mosquitoes prevalent in India which in turn could be helpful in devising appropriate approaches towards malaria control and prevention. The app can be modified as per future requirements.

High efficacy of microbial larvicides for malaria vectors control in the city of Yaounde Cameroon following a cluster randomized trial

The rapid expansion of insecticide resistance and outdoor malaria transmission are affecting the efficacy of current malaria control measures. In urban settings, where malaria transmission is focal and breeding habitats are few, fixed and findable, the addition of anti-larval control measures could be efficient for malaria vector control. But field evidences for this approach remains scarce. Here we provide findings of a randomized-control larviciding trial conducted in the city of Yaoundé that support the efficacy of this approach. A two arms random control trial design including 26 clusters of 2 to 4 km2 each (13 clusters in the intervention area and 13 in the non-intervention area) was used to assess larviciding efficacy. The microbial larvicide VectoMax combining Bacillus thuringiensis var israelensis (Bti) and Bacillus sphaericus in a single granule was applied every 2 weeks in all standing water collection points. The anopheline density collected using CDC light traps was used as the primary outcome, secondary outcomes included the entomological inoculation rate, breeding habitats with anopheline larvae, and larval density. Baseline entomological data collection was conducted for 17 months from March 2017 to July 2018 and the intervention lasted 26 months from September 2018 to November 2020. The intervention was associated with a reduction of 68% of adult anopheline biting density and of 79% of the entomological inoculation rate (OR 0.21; 95% CI 0.14–0.30, P < 0.0001). A reduction of 68.27% was recorded for indoor biting anophelines and 57.74% for outdoor biting anophelines. No impact on the composition of anopheline species was recorded. A reduction of over 35% of adult Culex biting densities was recorded. The study indicated high efficacy of larviciding for reducing malaria transmission intensity in the city of Yaoundé. Larviciding could be part of an integrated control approach for controlling malaria vectors and other mosquito species in the urban environment.

Residual malaria of Atlantic Forest systems and the influence of anopheline fauna.

In Brazil, the Amazon region comprises 99.5% of the reported malaria cases. However, another hotspot of the disease is the Atlantic Forest regions, with the sporadic occurrence of autochthonous human cases. In such context, this study sought to investigate the role of anopheline mosquitoes (Diptera: Culicidae) in the residual malaria transmission in Atlantic Forest areas. Two rural areas in the Espírito Santo state were the surveyed sites. Mosquitoes were captured using Shannon trap and CDC light traps and identified into species based on morphological characters. Ecological indexes (Shannon-Wiener diversity, Simpson's dominance, Pielou equability, and Sorensen similarity) were the tools used in the anopheline fauna characterization and comparison along with the two explored areas. The assessment of the sampling adequacy in the studied areas was possible through the generation of a species accumulation curve. A correlation test verified the influence of climatic variables on the anopheline species abundance. A total of 1471 female anopheline mosquitoes were collected from May 2019 to April 2020, representing 13 species. The species richness was higher in Valsugana Velha (hypo-endemic) than in Alto Caparaó (non-endemic). There was a significant variation in the species abundance between Valsugana Velha (n = 1438) and Alto Caparaó (n = 33). The most abundant species was Anopheles (Kerteszia) cruzii complex Dyar and Knab, 1908 representing 87% of the total anophelines collected. These results suggest that the Plasmodium spp. circulation in Brazilian Atlantic Forest areas occurs mainly due to the high frequency of Anopheles (K.) cruzii complex, considered the principal vector of simian and human malaria in the region.

Sialovirome of Brazilian tropical anophelines

Anophelinae is a widely dispersed Culicidae subfamily that may carry a unique virome. Here we herein report the set of viruses found in 323 salivary glands of 16 anopheline species sampled at Upper Pantanal, Chapada dos Guimarães National Park and Coxipó river basin, South Central Mato Grosso, Brazil, pooled (n = 11) and subjected to high throughput sequencing. Metagenomics revealed the presence of nine viral sequences belonging to novel viruses from seven viral families: Purunga is a putative novel orbivirus sharing 74% and 65% aa identity, respectively, with the VP1 and VP3 segments of Changuinola serogroup, Jaracatiá flavivirus shares 60% amino-acid (aa) identity with Aedes flavivirus. Coxipó dielmovirus and Chapada dielmovirus shared 51% and 39% aa identity with Merida virus. Coloiado-orthomyxo like virus is 57.1–64.8% identical at aa level to Aedes albonnulatus orthomyxo-like virus. Mujica picorna-like virus shares 49% aa identity with Flen picorna-like virus and Chiquitos virus is 50% similar to Ista virus, both from Picornavirales order. Cerrado partiti-like-virus shares 75–86% aa identity with Atrato partiti-like virus 2. We also found the S and L segments of Anopheles triannulatus orthophasmavirus (92% identity) in Anopheles lutzi from Chapada dos Guimarães. The identification of these putative novel viruses underscore the wide dispersion of viruses in culicid hosts contributing to extensions on mosquito virome descriptions.

Natural malaria infection in anophelines vectors and their incrimination in local malaria transmission in Darién, Panama.

BackgroundMore than 85% of the malaria cases in Panama occur in poor, rural and indigenous regions like Darien Province. Vector diversity, infection rate and spatial distribution are important entomological parameters of malaria transmission dynamics. Their understanding is crucial for the development of effective disease control strategies. The objective of this study was to determine the composition of Anopheles species, their natural infection rate and their geographic distribution to better understand the malaria transmission dynamics in Darién, Panama.MethodsAnophelines mosquitoes were captured during the rainy and dry season of 2016. We selected five communities where adult anophelines were collected using CDC light-traps, and through protective human-baited traps. Detection of natural infection and Plasmodium genotype were detected via nested PCR through the amplification of ssrRNA and the circumsporozoite protein gene (csp), respectively.ResultsA total of 1,063 mosquitoes were collected mosquitoes were collected for the detection of natural infection with Plasmodium spp. Nine Anophelines species were identified, with the predominant species being: An. (Nys.) darlingi (45.0%) and An. (Nys.) albimanus (42.6%). Natural infection in An. (Nys.) albimanus with P. vivax was detected in one mosquito pool from the community Pueblo Tortuga (0.6%), three from Marraganti (1.7%), two from Bajo Chiquito (1.1%) and three pools from Alto Playona 3 (1.7%). For An. (Nys.) darlingi mosquitoes, we detected seven positive pools from the community Bajo Chiquito (4.0%), two pools from Marraganti (1.1%) and two pools from Alto Playona (1.1%). The P. vivax allelic variant VK210 was detected in infected mosquitoes.ConclusionThe results from this study provide new information on the transmission dynamics associated with anophelines vectors in the Darién region. This is the first report of natural P. vivax infection in An. (Nys.) darlingi and its incrimination as a potential malaria vector in this region of Panama. Additional studies are necessary to expand our knowledge and determine crucial parameters in malaria transmission in Darién, which in turn will aid the National Malaria Program in attaining an adequate malaria control strategy towards malaria elimination.

Phylogenetic Complexity of Morphologically Identified Anopheles squamosus in Southern Zambia.

Simple SummaryDespite dramatic reductions in malaria cases in the catchment area of Macha Hospital, Choma District, Southern Province in Zambia, prevalence has remained near 1–2% by RDT for the past several years. To investigate residual malaria transmission in the area, this study focuses on the relative abundance, foraging behavior, and phylogenetic relationships of Anopheles squamosus specimens. Morphological identification, molecular verification of anopheline species, and blood meal source were determined on individual samples. Data from these collections supported earlier studies demonstrating An. squamosus to be primarily exophagic and zoophilic, allowing them to evade current control measures. The phylogenetic relationships generated from the specimens in this study supported the hypothesis of cryptic taxa among An. squamosus specimens, which further emphasizes the importance of molecular identification of vectors. The primarily exophagic behavior of An. squamosus in these collections also highlights that indoor vector control strategies will not be sufficient for elimination of malaria in southern Zambia.Despite dramatic reductions in malaria cases in the catchment area of Macha Hospital, Choma District, Southern Province in Zambia, prevalence has remained near 1–2% by RDT for the past several years. To investigate residual malaria transmission in the area, this study focuses on the relative abundance, foraging behavior, and phylogenetic relationships of Anopheles squamosus specimens. In 2011, higher than expected rates of anthropophily were observed among “zoophilic” An. squamosus, a species that had sporadically been found to contain Plasmodium falciparum sporozoites. The importance of An. squamosus in the region was reaffirmed in 2016 when P. falciparum sporozoites were detected in numerous An. squamosus specimens. This study analyzed Centers for Disease Control (CDC) light trap collections of adult mosquitoes from two collection schemes: one performed as part of a reactive-test-and-treat program and the second performed along a geographical transect. Morphological identification, molecular verification of anopheline species, and blood meal source were determined on individual samples. Data from these collections supported earlier studies demonstrating An. squamosus to be primarily exophagic and zoophilic, allowing them to evade current control measures. The phylogenetic relationships generated from the specimens in this study illustrate the existence of well supported clade structure among An. squamosus specimens, which further emphasizes the importance of molecular identification of vectors. The primarily exophagic behavior of An. squamosus in these collections also highlights that indoor vector control strategies will not be sufficient for elimination of malaria in southern Zambia.

Improved BioGents\xae Sentinel trap with heat (BGSH) for outdoor collections of Anopheline species in Burkina Faso and Mali, West Africa

BackgroundSince the late 1990s, malaria control programmes have relied extensively on mass bednet distribution and indoor residual spraying. Both interventions use pesticides and target mosquitoes coming indoors either to feed or to rest. Unfortunately, these intensified vector control campaigns have resulted in mosquito populations with high levels of resistance to most of the chemical compounds used against them and which are increasingly exophagic and exophillic, hence difficult to monitor indoors. Consequently, there is an urgent need for novel tools to sample outdoor anopheline populations for monitoring interventions and disease surveillance programmes.MethodologiesIn this study, we tested several modifications and configurations of the BioGents® Sentinel (BGS) trap, designed with the aim to increase its efficacy for sampling malaria vector species. Traps were used with chemical attractants and CO2, and the impacts of trap position, trap colour contrast combination and the addition of a heat source were tested in two studies conducted in the Sudano-Sahelian region of Burkina Faso and Mali.ResultsThe results show that of all the configurations tested, the addition of a heat source to the BGS trap with the original colour combination and an upward positioning resulted in a 1.8- and 5.9-fold increase in host-seeking Anopheles gambiae (s.l.) females in the experiments performed in Burkina Faso and Mali, respectively. BGS with heat traps, referred to as BGSH traps, captured An. gambiae (s.l.), An. pharoensis, An. coustani, Culex and Mansonia spp. Importantly, the results suggest that their efficacy does not depend on the close proximity of nearby hosts in houses.ConclusionsThe results suggest that BGSH traps can be an effective scalable tool for sampling outdoor anopheline vector populations. Further developments enabling CO2 and heat generation for longer periods of time would further improve the trap’s versatility for large-scale surveillance programmes.Graphical

Surveillance of Mosquitoes (Diptera, Culicidae) in Kyiv, Ukraine Between 2013 and 2017.

For effective control of vector-borne diseases and control of nuisance-biting insects, it is important to know which species are present and their relative abundance. In this study, we report data from a State-supported mosquito surveillance program in Kyiv, Ukraine's capital city. The surveillance identified 29 different species: 24 Culicines and 5 Anopheline species. Culicine mosquitoes included 17 in the genus Aedes, 3 Culex, 3 Culiseta, and 1 Mansonia species. The relative abundance of each genera was consistent in years 2014, 2015, and 2016; namely Aedes>Culex>Anopheles. In 2017, Aedes and Culex mosquitoes were approximately the same, predominating over Anopheles. A declining trend in the numbers of mosquitoes collected from 2013 to 2017 has not only several potential explanations, including increased urbanization and more effective control, but also may reflect changes in surveillance efforts.