Mosquito species distribution and larval breeding habitats with taxonomic identification of Anopheline mosquitoes in Korea

In 2005, adult and larval mosquito surveillance was conducted at selected sites in Korea to associate larval habitats with species distribution of mosquitoes of the Anopheles Hyrcanus Group (An. sinensis, An. lesteri, An. pullus, An. belenrae and An. kleini) and other mosquito species. Anopheles specimens belonging to the Anopheles Hyrcanus Group were identified to species level by molecular confirmation using the internal transcribed spacer (ITS)‐2 within nuclear ribosomal (r)DNA. A total of 6644 mosquitoes from resting and light trap collections (4451; 67.0%) and larval collections (2193; 33.0%) comprising 32 species and nine genera (Culex [11], Anopheles [8], Ochlerotatus [5], Aedes [3], Armigeres [1], Coquillettidia [1], Mansonia [1], Tripteroides [1] and Lutzia [1]) were collected. Larval habitats were characterized into 14 categories. Of a total of 4534 Anopheles spp. collected (3766 resting and light trap collections and 768 larval collections), Anopheles sinensis (3194; 70.4%) was the most frequently captured, followed by An. kleini (813; 17.9%), An. pullus (299; 6.6%) and An. belenrae (129; 2.8%). Four species of Anopheles (An. lesteri, An. sineroides, An. koreicus and An. lindesayi) were infrequently collected (<3.0%) at all sites surveyed by all methods of collection. Anopheles kleini, An. pullus and An. belenrae were collected in greater proportions in malaria high‐risk areas north of Seoul, and were infrequently collected in other parts of Korea, where An. sinensis was the predominant Anopheles spp. captured. A total of 2110 culicine mosquitoes (685 adult collections and 1425 larval collections) comprising 24 species and eight genera were collected.     

Distribution of overwintering mosquitoes (Diptera: Culicidae) in grassy fields in the Republic of Korea, 2007–2008

Overwintering mosquitoes in a grassy field were surveyed at 13 different localities from December 2007 to February 2008 in the Republic of Korea (ROK). Mosquitoes were collected by putting a white colored tent on the grass, without the base, and heating the inside of the tent. A total of 784 female adults comprising seven species were collected during the study period, including 395 (45.6%) Anopheles pullus, 338 (43.2%) An. sinensis, 21 (2.7%) An. belenrae, 2 (0.3%) An. kleini, 43 (5.5%) Culex tritaeniorhynchus, 20 (2.5%) Cx. inatomii, and 1 (0.3%) Cx. pipiens complex. The mosquitoes showed different overwintering distributions by temperature zone and locality. Predominant species, the malaria vectors An. pullus and An. sinensis, showed different overwintering distributions. Anopheles pullus had a 90.9% collection rate except for Youngju and an average – 0.6°C temperature between December and February. Anopheles pullus showed >50% collection rate in most regions maintained below an average of ?1.4°C, and the rate of An. sinensis showed more than 50% collection rate in all regions maintained at an average above ?1.1°C. Other species showed regional characteristics; An. belenrae was distributed in the northwest part of the ROK, and An. kleini was collected rarely at one locality in the middle part of ROK. Culex tritaeniorhynchus, a Japanese encephalitis virus vector, was distributed in the southern and costal region and maintained at an above average of ?1.0°C. Culex inatomii was collected on the coastal region of the ROK.     

Distribution and larval habitats of Anopheles species in northern Gyeonggi Province,Republic of Korea

A total of 180 larval collection sites (e.g., rice paddies, marshes, ground pools, ponds, stream margins, and irrigation and drainage ditches) was surveyed within a 2 km radius from Warrior Base training area, 5 km south of Panmunjeom (Joint Security Area, demilitarized zone), Gyeonggi Province, Republic of Korea (ROK), from May through October, 2007 to characterize larval habitat distributions of members of the Anopheles Hyrcanus Group (An. sinensis, An. lesteri, An. pullus, An. belenrae, An. kleini, and An. sineroides). A total of 5,859 anopheline larvae was collected from 84.4% of the sites surveyed, of which 4,071 were identified to species by polymerase chain reaction (PCR) using the ribosomal DNA internal transcribed spacer 2 (rDNA ITS2). Anopheles sinensis (52.6%) was the most frequently collected, followed by An. kleini (29.4%), An. sineroides (9.8%), An. pullus (6.7%), An. belenrae (1.1%), and An. lesteri (0.5%). Anopheles pullus and An. kleini were collected in greater proportions in May and from May – July, respectively. Few An. sinensis were collected from May – June, but it was the predominant species collected by August, and accounted for >80% of all larvae from September – October. Anopheles kleini was found in all habitats sampled; however, it was collected most frequently in young growth rice paddies, while An. sinensis was collected more frequently in mature and post‐harvest paddies. Anopheles pullus was associated with pre‐cultivated rice paddies, including water‐filled tire ruts left from the previous fall's harvest.     

Bionomics of Anopheles spp. (Diptera: Culicidae) in a malaria endemic region of Sukabumi,West Java,Indonesia

A 15‐month bionomic study of Anopheles species was conducted in two ecologically distinct villages (coastal and upland) of Sukabumi District, West Java, Indonesia from June 2006 to September 2007. Mosquitoes were captured using human‐landing collections at both sites. During the study, a total of 17,100 Anopheles mosquitoes comprising 13 Anopheles species were caught: 9,151 at the coastal site and 7,949 at the upland site. Anopheles barbirostris, Anopheles maculatus, and Anopheles vagus were the predominant species caught at the coastal site, and Anopheles aconitus, Anopheles barbirostris, and An. maculatus predominated in the upland site. Overall, species were exophagic at both sites, but there was variation between species. Anopheles aconitus was endophagic at the coastal site, exophagic at the upland site, collected most often in April 2007 and had a peak landing time between 22:00 and 23:00. Anopheles sundaicus was only collected at the coastal site, exophagic, collected most often in October 2006, and had a peak landing time between 19:00 and 20:00. Potential malaria vector species such An. aconitus, An. maculatus, and An. sundaicus were present throughout the year. None of the 7,770 Anopheles tested using CSP‐ELISA were positive for malaria, although the risk for malaria outbreaks in Sukabumi district remains high.     

Malaria Vector Surveillance in Ganghwa-do, a Malaria-Endemic Area in the Republic of Korea

We investigated the seasonality of Anopheles mosquitoes, including its species composition, density, parity, and population densities of mosquitoes infected with the parasite in Ganghwa-do (Island), a vivax malaria endemic area in the Republic of Korea. Mosquitoes were collected periodically with a dry-ice-tent trap and a blacklight trap during the mosquito season (April-October) in 2008. Anopheles sinensis (94.9%) was the most abundant species collected, followed by Anopheles belenrae (3.8%), Anopheles pullus (1.2%), and Anopheles lesteri (0.1%). Hibernating Anopheles mosquitoes were also collected from December 2007 to March 2008. An. pullus (72.1%) was the most frequently collected, followed by An. sinensis (18.4%) and An. belenrae (9.5%). The composition of Anopheles species differed between the mosquito season and hibernation seasons. The parous rate fluctuated from 0% to 92.9%, and the highest rate was recorded on 10 September 2008. Sporozoite infections were detected by PCR in the head and thorax of female Anopheles mosquitoes. The annual sporozoite rate of mosquitoes was 0.11% (2 of 1,845 mosquitoes). The 2 mosquitoes that tested positive for sporozoites were An. sinensis. Malarial infections in anopheline mosquitoes from a population pool were also tried irrespective of the mosquito species. Nine of 2,331 pools of Anopheles mosquitoes were positive. From our study, it can be concluded that An. sinensis, which was the predominant vector species and confirmed as sporozoite-infected, plays an important role in malaria transmission in Ganghwa-do.     

Mosquito diversity (Diptera: Culicidae) and medical importance,in a bird sanctuary inside the flooded forest of Prek Toal,Cambodia

Wild birds have been identified as reservoirs of many pathogens transmissible to humans through bird-biting mosquitoes. Understanding the diversity of vector fauna in bird-conservation areas, is a cornerstone to later develop early detection systems and reduce the risk of emerging disease incidence. Prek Toal is a bird Sanctuary in the Tonle-Sap area in Cambodia hosting the largest migratory bird colony of southeast Asia. The Culicidae fauna was investigated during both dry and rainy seasons. In total, 8,224 mosquitoes from 6 genera and 25 species were collected. The two most represented genera were Anopheles (8 species) and Culex (11 species), while the dominant species was Culex vishnui (58.8%). During the rainy season, 1.6 fold more mosquitoes were caught and the number of species doubled. One of the most notable feature is the absence of any Aedinii mosquito which could be attributed to the low human density around the area, and the large populations of migratory birds. A total of 13 species from 3 genera are reported to be of medical importance. 6 medically-important species were collected in all 3 sites, and are known to bite both wild birds and humans: An. barbirostris, An. campestris, Cx. bitaeniorhynchus, Cx. vishnui, Ma. indiana and Ma. uniformis. One of the most important feature is the over-abundance of Culex vishnui, largely dominant in Prek Toal throughout the seasons and the sites, and being present everywhere in Cambodia. This finding stresses its potentiality to act as bridge vectors in the case of new disease emergence.     

Biting patterns and host preference of Anopheles epiroticus in Chang Island,Trat Province,eastern Thailand

A study of species diversity of Anopheles mosquitoes, biting patterns, and seasonal abundance of important mosquito vectors was conducted in two villages of Chang Island, Trat Province, in eastern Thailand, one located along the coast and the other in the low hills of the central interior of the island. Of 5,399 captured female anophelines, 70.25% belong to the subgenus Cellia and remaining specimens to the subgenus Anopheles. Five important putative malaria vectors were molecularly identified, including Anopheles epiroticus, Anopheles dirus, Anopheles sawadwongporni, Anopheles maculatus, and Anopheles minimus. Anopheles epiroticus was the most commonly collected species in the coastal site, whereas An. dirus was found to be most abundant in the forest‐hill site. From both locations, a greater number of mosquitoes was collected during the dry season compared to the wet. Anopheles epiroticus showed greater exophagic and zoophilic behavior with the highest blood feeding densities occurring between 18:00 and 19:00. In contrast, An. dirus demonstrated an activity peak between midnight and 01:00. We conclude that An. epiroticus and An. dirus, in coastal and inland areas, respectively, appear to be the most epidemiologically important malaria vectors on Chang Island. As no studies of vector competency specific to Chang Island have been conducted, our conclusions that these two species play a primary role in malaria transmission are based on evidence from other localities in Thailand and mainland Southeast Asia. This information serves as a basis for designing improved vector control programs that target specific species, and if integrated with other interventions could result in the elimination of malaria transmission on the island.     

A recent survey of mosquito fauna in Guandong Province, southern China, with a review of past records

Abstract The southern province of Guangdong has long been subject to endemic mosquito‐borne diseases. In recent years, this region of China has experienced rapid, extensive economic development involving environmental change, making much of the scant knowledge of its mosquito fauna obsolete. This paper reviews previous mosquito surveys, some of which may be too old to be of relevance to present‐day conditions, and presents the results of a recent survey of adult and immature mosquitoes. The main vectors of mosquito‐borne diseases endemic to the area, such as dengue and Japanese encephalitis virus, develop in container habitats. A three‐year survey was carried out, between 2004 and 2006, of 4131 breeding containers in residential areas and in open, sparsely populated areas, of which approximately 50% were positive for the presence of mosquitoes, and 10 156 larvae and pupae were collected and identified. Twelve species were found in both residential and sparsely populated areas: Aedes albopictus (Skuse) (Diptera: Culicidae), Ae. lineatopennis (Ludlow), Ae. vexans (Meigen), Tanakaius togoi (Theobald), Culex barraudi Edwards, Cx dispectus Bram, Cx malayi (Leicester), Cx pallidothorax Theobald, Cx quinquefasciatus Say, Cx sitiens Wiedemann, Lutzia fuscanus Wiedemann and Tripteroides bambusa (Yamada). Armigeres subalbatus (Coquillett) was found only in containers in villages, whereas Ae. macfarlanei Edwards, Cx mimeticus Noé, Cx sinensis Theobald, Cx vegans Wiedemann, Cx wilfredi Colless and Mansonia uniformis (Theobald) were found only in non‐ or sparsely populated areas. In residential areas, the rank order of most common species, as measured by the proportion of containers colonized, was Ae. albopictus > Cx quinquefasciatus > Lu. fuscanus, whereas in sparsely populated areas the rank order was Cx quinquefasciatus > Ae. albopictus > Lu. fuscanus. Light traps in non‐ or sparsely populated areas caught 5995 adult mosquitoes of 25 species, some of which are not container breeders. The most common species were: Anopheles sinensis Wiedemann, An. maculatus Theobald, An. minimus Theobald, Ta. togoi, Cx bitaeniorhynchus Giles, Cx malayi, Cx quinquefasciatus, Cx sinensis Theobald, Cx sitiens, Cx tritaeniorhynchus Giles and Lu. fuscanus. It is noteworthy that nine species caught had not been previously recorded in Guangdong Province, highlighting the deficient knowledge of the current composition and distribution of the mosquito fauna of this part of China.     

Diversity of Anopheles species and trophic behavior of putative malaria vectors in two malaria endemic areas of northwestern Thailand

We determined the species diversity, blood‐feeding behavior, and host preference of Anopheles mosquitoes in two malaria endemic areas of Tak (Mae Sot District) and Mae Hong Son (Sop Moei District) Provinces, located along the Thai border with Myanmar, during a consecutive two‐year period. Anopheline mosquitoes were collected using indoor and outdoor human‐landing captures and outdoor cow‐baited collections. Mosquitoes were initially identified using morphological characters, followed by the appropriate multiplex AS‐PCR assay for the identification of sibling species within Anopheles (Cellia) complexes and groups present. Real‐time PCR was performed for parasite‐specific detection in mosquitoes (Plasmodium spp. and Wuchereria bancrofti). A total of 7,129 Anopheles females were captured, 3,939 from Mae Sot and 3,190 from Sop Moei, with 58.6% and 37% of all anophelines identified as An. minimus, respectively. All three malaria vector complexes were detected in both areas. One species within the Minimus Complex (An. minimus) was present along with two related species in the Funestus Group, (An. aconitus, An. varuna), two species within the Dirus Complex (An. dirus, An. baimaii), and four species within the Maculatus Group (An. maculatus, An. sawadwongporni, An. pseudowillmori, and An. dravidicus). The trophic behavior of An. minimus, An. dirus, An. baimaii, An. maculatus, and An. sawadwongporni are described herein. The highest An. minimus densities were detected from February through April of both years. One specimen of An. minimus from Mae Sot was found positive for Plasmodium vivax.     

Malaria and the Anopheles mosquitoes of Tajikistan

Surveys of Anopheles mosquitoes were conducted in urban, rural, and natural areas of Tajikistan to obtain updated information on their distributions, especially in southern districts of the country where malaria is a prevalent disease. Nine species of Anopheles are found in Tajikistan. Anopheles superpictus, An. claviger, An. hyrcanus, and An. pulcherrimus are the most widespread and abundant species. Investigations in northern Tajikistan confirmed the presence of An. artemievi and the absence of An. martinius, both members of the An. maculipennis complex of malaria vectors. Anopheles barianensis, An. lindesayi, and An. marteri sogdianus, species previously recorded in the country, were not encountered during our surveys. The history of Anopheles and malaria research in Tajikistan is reviewed and bionomical and distributional information is provided for each of the nine species.     

Mosquito community composition and abundance at contrasting sites in northern South Africa, 2014–2017

Most data on species associations and vector potential of mosquitoes in relation to arboviral infections in South Africa date back from the 1940s to late 1990s. Contextual information crucial for disease risk management and control, such as the sampling effort, diversity, abundance, and distribution of mosquitoes in large parts of South Africa still remains limited. Adult mosquitoes were collected routinely from two horse farms in Gauteng Province; two wildlife reserves in Limpopo Province, at Orpen Gate in Kruger National Park (KNP) and Mnisi Area in Mpumalanga Province between 2014–2017, using carbon dioxide‐baited light and tent traps. Mosquito diversity and richness are greater in untransformed natural and mixed rural settings. In untransformed wilderness areas, the most dominant species were Culex poicilipes, Anopheles coustani, and Aedes mcintoshi, while in mixed rural settings such as the Mnisi area, the two most abundant species were Cx. poicilipes and Mansonia uniformis. However, in peri‐urban areas, Cx. theileri, Cx. univittatus, and Cx. pipiens sensu lato were the most dominant. Aedes aegypti, Ae. mcintoshi, Ae. metallicus, Ae. vittatus, Cx. pipiens s.l., Cx. theileri, and Cx. univittatus had the widest geographical distribution in northern South Africa. Also collected were Anopheles arabiensis and An. vaneedeni, both known malaria vectors in South Africa. Arbovirus surveillance and vector control programs should be augmented in mixed rural and peri‐urban areas where the risk for mosquito‐borne disease transmission to humans and domestic stock is greater.     

Maintenance of mosquito vectors: effects of blood source on feeding,survival, fecundity,and egg hatching rates

Artificial membrane‐feeding techniques have replaced direct feeding on animals for the maintenance of malaria and arbovirus vectors in many laboratories. Membrane feeding facilitates controlled experimentation of pathogen transmission during mosquito feeding. Sheep blood is commonly used due to its availability and low cost. We evaluated the impact of blood source (human, guinea pig, sheep, and hamster via direct feeding) on feeding rates, adult survival, fecundity, hatching rates, and developmental times for five species of laboratory‐colonized mosquitoes (Anopheles dirus, An. cracens, An. minimus, An. sawadwongporni, and Ae. aegypti). We found that feeding rates differ among blood sources within mosquito species. Survival, fecundity, and hatching rates were lower in all Anopheles species and Ae. aegypti after membrane feeding on sheep blood. Survival rates seven days post‐feeding on sheep blood were significantly lower (P<0.05) for An. dirus (84.2%), An. minimus (67.2%), An. sawadwongporni (51.5%), and An. cracens (35.5%) relative to other blood sources. An. minimus and An. sawadwongporni laid no eggs by seven days post‐feeding with sheep blood, while An. dirus and An. cracens produced significantly fewer numbers of eggs and demonstrated significantly lower hatching rates relative to what was observed with the other blood sources. These findings support the conclusion that sheep blood is not a suitable blood source for laboratory rearing of Anopheles spp.     

Seasonal prevalence of mosquitoes collected from light traps in the Republic of Korea, 2007

Adult mosquito surveillance was conducted using New Jersey light traps from 1 May to 15 October during 2007 at 14 US Army and three Air Force installations and two US Army training sites located in six provinces in the Republic of Korea to determine threshold levels to initiate pesticide applications based on trap indices. A total of 24 013 adult mosquitoes (19 763 [82.3%] females and 4250 [17.1%] males) comprising 16 species (including five members of the Anopheles Hyrcanus Group) and belonging to seven genera were collected. Females of the most commonly collected species were members of the Anopheles Hyrcanus Group (52.1%), followed by, Culex tritaeniorhynchus Giles (20.6%), Culex pipiens Coquillett (13.8%), and Aedes vexans nipponii (Theobald) (11.6%). Trap indices varied widely for species over their range, due in part, to geographical distribution and degree of association with urban or rural communities.     

Diversity,distribution, abundance,and feeding pattern of tropical ornithophilic mosquitoes

Bird–biting mosquitoes act as bridge vectors of diverse pathogens of emerging infectious diseases. In this study, we report for the first time the abundance, diversity, distribution, and feeding pattern of bird‐biting mosquitoes on an island where avifaunal diversity is rich. Monthly mosquito collections were done at six different habitats in three different climatic zones using bird‐baited traps over a year. Collected mosquitoes were identified using morphological and molecular tools. A total of 2,655 bird‐biting mosquitoes of eight genera and 25 species were identified. Of these, 52% were Culex species, which represents 35% of the Culex species in the country. The most abundant species were Culex sitiens, Cx. pseudovishnui, Cx. nigropunctatus and Cx. quinquefasciatus, whereas the latter two were common to all habitats. The highest abundance was reported in lowland forests (49.6%), while it was lowest in highland forests (22.3%). Highest species similarity was reported from highland forests. Seasonal variations of the most abundant species were significantly different in selected habitats (p< 0.05). Two distinct biting peaks were identified, from 06:00 to 21:00 and 22:00 to 02:00. The biting nature of identified ornithophilic mosquitoes suggests the potential vector status of these mosquitoes.     

Flaviviral disease mosquito vector surveillance in Incheon Metropolitan City and the Hwaseong area,Gyeonggi‐Do,Republic of Korea,in 2015

Owing to global climate change, the global resurgence of vector‐borne infectious diseases and their potential to inflict widespread casualties among human populations has emerged as a pivotal burden on public health systems. In this study, the prevalence of flaviviral diseases transmitted by mosquitoes, and their target vector diversity, abundance, and distribution was investigated to enable the mapping of hotspots for these diseases. For the surveillance of the vector mosquitoes carrying flaviviruses during April to November 2015, female mosquitoes were collected to study whether they carried pathogens from abroad at seven locations in Incheon Metropolitan City (Incheon) as a typical urban area and Hwaseong‐si (= city, Hwaseong) of Gyeonggi‐do (= province) as a rural area. A total of 15 species belonging to seven genera (29,102 female mosquitoes) were collected with black‐light and BG‐Sentinel? traps at a collection rate of 260 per trap/night from whole collection locations. The most collected mosquito species in Incheon were Aedes vexans nipponii (species ratio (SR), 29.9%) and the Culex pipiens complex (SR, 28.8%), followed by Anopheles sinensis s.l. (SR, 27.9%) and Ochlerotatus koreicus (SR, 7.1%). From the results of viral RNA detection, five flaviviruses were found in 20,981 individuals (excluding An. sinensis; 696 pools) in the Cx. pipiens complex and Ae. vexans nipponii. Three Japanese encephalitis virus (JEV)‐positive pools were from the Cx. pipiens complex, a Chaoyang virus pool was found from Ae. vexans nipponii, and the remaining unidentified flavivirus pool was from Cx. pipiens. The three JEV‐positive pools were phylogenetically grouped as genotype V. The results of our study demonstrate that enhanced monitoring and long‐term surveillance of these vector viruses are of great public health importance.     

Daily and seasonal variation in the activity of potential vector mosquitoes

In the course of season 2010 (May–September), three 2-day trappings of female mosquitoes were carried out at two sites in order to determine the daily activity of the common mosquito species (e.g. species from genus Culex, Aedes, Ochlerotatus, Anopheles, etc.) in the area. CDC light traps filled with CO₂ and placed at a height of 1 m were used to trap individuals, and were sampled every 2 h. A total of 19,604 female mosquitoes from 20 identifiable species were trapped: 7,549 at Sedlec and 12,055 at Kančí obora. The activity of the major species of mosquitoes in South Moravia differed throughout the course of the day. Calamity species of the genus Aedes and Ochlerotatus remained active throughout the day and night, but with different course. Aedes vexans, Ae. cinereus, and Ae. rossicus were most active in the late afternoon and highly active for most of the night. Ochlerotatus sticticus was captured most often in the afternoon, and its abundance decreased rapidly before sunset. The activity of Oc. cantans s.l. (Oc. cantans + Oc. annulipes) females varied little during the day and night. The daily activity for the main vectors of West Nile virus, Culex pipiens and Cx. modestus, were totally different from that of other species. Cx. pipiens females showed significant night activity, while Cx. modestus was most active in the evening. Nighttime activity was also observed in female mosquitoes of the genus Anopheles.     

Anopheline species and their Plasmodium infection status in Aligarh,India

Malaria is a global issue and India contributes substantially to global malaria incidence. Information related to malaria vectors is very limited in Aligarh. The environmental and climatological situations permit the continual breeding of vectors in permanent breeding sites. This study was designed with the aim to screen all the anophelines species and possible malaria vectors in three different localities of Aligarh. Anopheles mosquitoes were collected from three different localities (Fort, Jalali and Tappal) during peak malaria transmission season (July to November) by using mouth aspirator and CDC light traps. Enzyme-linked immunosorbent assay (ELISA) was done to detect Plasmodium falciparum, Plasmodium vivax-210 and P. vivax-247 circumsporozoite proteins (CSP) from the collected female species. A total of 794 female anopheline mosquitoes belonging to 7 species were collected by different methods. Circumsporozoite protein–enzyme-linked immunosorbent assay was performed with 780 anopheline mosquitoes out of which 13 mosquitoes were positive in CSP–ELISA. Thus, the overall infection rate was 1.66% (13/780). Four (0.51%) mosquitoes belonging to three species were positive for P. falciparum, 7 (0.89%) mosquitoes belonging to three species were positive for VK 210 and 2 (0.25%) mosquitoes belonging to Anopheles culicifacies and Anopheles stephensi species were positive for VK 247. No mixed infection was found in this study. According to species, the highest infection rate was observed in An. culicifacies (7/288, 2.43%) followed by An. stephensi (2.40%) and Anopheles annularis (1.98%). An. culicifacies and An. stephensi were previously incriminated as malaria vectors in Aligarh. There was, however, no previous report in favor of infections in An. annularis in Aligarh. The on-going Malaria Control Program in India needs up to date information on malaria vectors. A major challenge is the lack of knowledge about vectors and their role in malaria transmission. Findings of this study suggested that in the absence of major malaria vectors there is a possibility that other Anopheles species may have been playing a role in malaria transmission in Aligarh.     

Wide recognition of Culex pipiens and lack of detection of Culex torrentium through biomolecular differentiation of mosquitoes in the Emilia‐Romagna region,Northern Italy

The Culex pipiens complex includes species with reported differences in vector competence for arthropod‐borne viruses, many of which are of significant importance to human health such as the West Nile virus and the Sindbis virus. This group of mosquitoes is difficult to distinguish morphologically; particularly as adult females. In Europe, the two species of the complex, Culex pipiens Linnaeus 1758 and Culex torrentium Martini 1925, are often found sympatrically. With the aim to characterize the presence and spread of both species in the Emilia‐Romagna region, Northern Italy, mosquitoes of the complex – collected during the West Nile virus surveillance plans – were tested by multiplex real‐time PCR for the detection of the two species Cx. pipiens and Cx. torrentium. A total of 24 165 mosquitoes, collected between 2012 and 2014 from 105 sites, and sorted in 204 pools, were tested. All tested pools were found to be composed of Cx. pipiens, whereas Cx. torrentium was not detected. These results indicate a likely absence of Cx. torrentium mosquitoes within the surveyed territory, whereas Cx. pipiens is widely distributed in the area mentioned. This is in line with previous reports, which describe a pre‐alpine distribution of Cx. torrentium in Italy.     

Whole‐genome sequencing reveals absence of recent gene flow and separate demographic histories for Anopheles punctulatus mosquitoes in Papua New Guinea

Anopheles mosquitoes are the vectors of several human diseases including malaria. In many malaria endemic areas, several species of Anopheles coexist, sometimes in the form of related sibling species that are morphologically indistinguishable. Determining the size and organization of Anopheles populations, and possible ongoing gene flow among them is important for malaria control and, in particular, for monitoring the spread of insecticide resistance alleles. However, these parameters have been difficult to evaluate in most Anopheles species due to the paucity of genetic data available. Here, we assess the extent of contemporary gene flow and historical variations in population size by sequencing and de novo assembling the genomes of wild‐caught mosquitoes from four species of the Anopheles punctulatus group of Papua New Guinea. Our analysis of more than 50 Mb of orthologous DNA sequences revealed no evidence of contemporary gene flow among these mosquitoes. In addition, investigation of the demography of two of the An. punctulatus species revealed distinct population histories. Overall, our analyses suggest that, despite their similarities in morphology, behaviour and ecology, contemporary sympatric populations of An. punctulatus are evolving independently.