Culex Gelidus Research Articles

Demographic inference from the mt-DNA COI gene and wing geometry of Culex gelidus (Diptera: Culicidae), an important vector of Japanese encephalitis in Thailand

Culex gelidus (Diptera: Culicidae), an important vector of the Japanese encephalitis virus (JEV), contributes to human viral encephalitis in many Asian countries, including Thailand. This study represents the first investigation of the demographic patterns of Cx. gelidus populations in Thailand using cytochrome c oxidase subunit I (COI) gene analysis and wing geometric morphometrics (GM). Mosquitoes were collected from 10 provinces across six regions of Thailand in 2022. Analysis of the COI sequences (n = 182) indicated high haplotype diversity (0.882) and low nucleotide diversity (0.006), with 72 haplotypes identified. The haplotype network demonstrated no profound splits among the geographic populations. Neutral tests, including Tajima's D and Fu's Fs, displayed negative values, with a significant result observed for Fu's Fs (−33.048, p < 0.05). The mismatch distribution analysis indicated that the population does not statistically deviate from a model of sudden population expansion (SSD = 0.010, p > 0.05; Rg = 0.022, p > 0.05). The estimations suggest that the Cx. gelidus population in Thailand began its expansion approximately between 459,243 and 707,011 years ago. The Mantel test showed no significant relationship between genetic and geographic distances (r = 0.048, p > 0.05). Significant phenotypic differences (based on wing shape) were observed among most populations. Additionally, in this study, we found no significant relationships between phenotypic and genetic distances (r = 0.250, p > 0.05). Understanding the genetic and morphological dynamics of Cx. gelidus is vital for developing targeted surveillance and vector control measures. This knowledge will also help to predict how future environmental changes might affect these populations, thereby informing long-term vector management strategies.

Detection of Wuchereria bancrofti DNA in wild caught vector and non-vector mosquitoes: implications for elimination of lymphatic filariasis.

Transmission Assessment Survey (TAS) is the WHO recommended method used for decision-making to stop or continue the MDA in lymphatic filariasis (LF) elimination programme. The WHO has also recommended Molecular Xenomonitoring (MX) of LF infection in vectors as an adjunct tool in settings under post-MDA or validation period. Screening of non-vectors by MX in post-MDA / validation settings could be useful to prevent a resurgence of LF infection, as there might be low abundance of vectors, especially in some seasons. In this study, we investigated the presence of LF infection in non-vectors in an area endemic for LF and has undergone many rounds of annual MDA with two drugs (Diethylcarbamazine and Albendazole, DA) and two rounds of triple drug regimens (Ivermectin + DA). Mosquitoes were collected from selected villages of Yadgir district in Karnataka state, India, during 2019. A total of 680 female mosquitoes were collected, identified morphologically by species and separated as pools. The female mosquitoes belonging to 3 species viz., Anopheles subpictus, Culex gelidus and Culex quinquefaciatus were separated, pooled, and the DNA extracted using less expensive method and followed by LDR based real-time PCR assay for detecting Wuchereria bancrofti infection in vector as well as non-vector mosquitoes. One pool out of 6 pools of An. subpictus, 2 pools out of 6 pools of Cx. gelidus, and 4 pools out of 8 pools of Cx. quinquefaciatus were found to be positive for W. bancrofti infection by RT-PCR. The infection rate in vectors and non-vectors was found to be 1.8% (95% CI: 0.5-4.2%) and 0.9% (95% CI:0.2-2.3%), respectively. Our study showed that non-vectors also harbour W. bancrofti, thus opening an opportunity of using these mosquitoes as surrogate vectors for assessing risk of transmission to humans in LF endemic and post MDA areas.

Characterizing the Wolbachia infection in field-collected Culicidae mosquitoes from Hainan Province, China

BackgroundMosquitoes are vectors of many pathogens, such as malaria, dengue virus, yellow fever virus, filaria and Japanese encephalitis virus. Wolbachia are capable of inducing a wide range of reproductive abnormalities in their hosts, such as cytoplasmic incompatibility. Wolbachia has been proposed as a tool to modify mosquitoes that are resistant to pathogen infection as an alternative vector control strategy. This study aimed to determine natural Wolbachia infections in different mosquito species across Hainan Province, China.MethodsAdult mosquitoes were collected using light traps, human landing catches and aspirators in five areas in Hainan Province from May 2020 to November 2021. Species were identified based on morphological characteristics, species-specific PCR and DNA barcoding of cox1 assays. Molecular classification of species and phylogenetic analyses of Wolbachia infections were conducted based on the sequences from PCR products of cox1, wsp, 16S rRNA and FtsZ gene segments.ResultsA total of 413 female adult mosquitoes representing 15 species were identified molecularly and analyzed. Four mosquito species (Aedes albopictus, Culex quinquefasciatus, Armigeres subalbatus and Culex gelidus) were positive for Wolbachia infection. The overall Wolbachia infection rate for all mosquitoes tested in this study was 36.1% but varied among species. Wolbachia types A, B and mixed infections of A × B were detected in Ae. albopictus mosquitoes. A total of five wsp haplotypes, six FtsZ haplotypes and six 16S rRNA haplotypes were detected from Wolbachia infections. Phylogenetic tree analysis of wsp sequences classified them into three groups (type A, B and C) of Wolbachia strains compared to two groups each for FtsZ and 16S rRNA sequences. A novel type C Wolbachia strain was detected in Cx. gelidus by both single locus wsp gene and the combination of three genes.ConclusionOur study revealed the prevalence and distribution of Wolbachia in mosquitoes from Hainan Province, China. Knowledge of the prevalence and diversity of Wolbachia strains in local mosquito populations will provide part of the baseline information required for current and future Wolbachia-based vector control approaches to be conducted in Hainan Province.Graphical

Worldwide transmission and infection risk of mosquito vectors of West Nile, St. Louis encephalitis, Usutu and Japanese encephalitis viruses: a systematic review

The increasing trend of mosquito-borne pathogens demands more accurate global estimations of infection and transmission risks between mosquitoes. Here, we systematically review field and laboratory studies to assess the natural field infection and experimental laboratory transmission risk in Culex mosquitoes. We studied four worldwide flaviviruses: West Nile, Usutu, Japanese encephalitis, and St. Louis encephalitis, belonging to the Japanese encephalitis Serocomplex (JES). The PRISMA statement was carried out for both approaches. The Transmission-Infection Risk of the diverse mosquito species for the different viruses was estimated through seven variables. We considered 130 and 95 articles for field and experimental approach, respectively. We identified 30 species naturally infected, and 23 species capable to transmit some of the four flaviviruses. For the JES, the highest Transmission-Infection Risk estimate was recorded in Culex quinquefasciatus (North America). The maximum Infection-Transmission Risk values for West Nile was Culex restuans, for Usutu it was Culex pipiens (Europe), for St. Louis encephalitis Culex quinquefasciatus (North America), and for Japanese encephalitis Culex gelidus (Oceania). We conclude that on a worldwide scale, a combination of field and experimental data offers a better way of understanding natural infection and transmission risks between mosquito populations.

Occurrence of Some Mosquito Species in Tha-Ma-Nya Village, Hpa-an Township, Kayin State in Myanmar

Among the numerous species of blood-sucking insects, mosquitoes stand out as most offensive to man and other warm-blooded animals. Mosquitoes were collected by indoor and outdoor CDC light traps in randomly selected household and larvae were collected in and around 3 Kilo meter radius from the study site. A total of 14 species, five genera under family Culicidae were recorded in Tha-ma-nya village. Recorded species included two species under genus Aedes, three species under genus Culex one species under the genus Armigeres, one species under the genus Toxorhynchities and 7 species under genus Anopheles. Species inentification was done according to different morphological keys. Altogether seven out of 14 recorded mosquito species were medically important vector species. Aedes aegypti and Ae. albopictus were primary and secondary vectors of dengue fever. Culex quinquefasciatus is a microfilariasis vector. The important vectors of Japanese encephalitis are Culex tritaeniorhynchus and Culex gelidus. High density of malaria vector Anopheles dirus was collected in outdoor light trap than An. minimus, although An. minimus was collected high density by indoor CDC light traps. Virus of Japanese encephalitis could transmit from rearing pig and ducks to persons living in the study area. Indoor clear water containers serve as the favorable places for living Aedes spp. Outdoor water storage concrete tanks provided stagnant water to breed Culex and Anopheles species. Water pools, rock pools and rice fields water were found high preferences for breeding of malaria vectors An. dirus and An. minimus larvae and An. dirus larvae was also collected from concrete tanks. Microfilariasis vector Cx. quinquefqsciatus and JE vector Cx. tritaeniorhynchus larvae were collected from polluted water pools and Dengue vector larvae as Aedes larvae were collected from metal drums, bago jars and concrete tanks. Status of the mosquito species composition in the Tha-ma-nya village provided database not only for further researches but also for giving good knowledge and practice in health workers for effective control of vector borne diseases.

Comparative study on the larvicidal effect of some ciliated protists on Culex gelidus, Culex tritaeniorhynchus, and Aedes aegypti in Sri Lanka

Rice fields in Sri Lanka create suitable breeding places for vector mosquitoes. Such sites provide habitats for diversified naturally occurring microbiota. Ciliated protists, Zoothamnium sp., Chilodonella sp., and Vorticella microstoma are among such microbiota found in vector mosquito habitats especially in rice field habitats in Sri Lanka. The present study was carried out to determine the comparative larvicidal effect of these ciliated protists collected from naturally infested mosquito larvae in some rice-field habitats in Kurunegala, Sri Lanka, against vector mosquito larvae. Vector mosquito larvae, Culex tritaeniorhynchus, and Culex gelidus were reared in the laboratory from field-collected water samples while Aedes aegypti mosquito larvae were reared using egg sheets, for the laboratory bioassays. V. microstoma showed the potential for infection and mortality of Cx. tritaeniorhynchus larvae (71.33% ± 5.23). Results revealed a minimum of 1000V. microstoma is required to kill a single third instar larva of Cx. tritaeniorhynchus at 69.60 ± 2.40h of exposure. Cx. gelidus larvae showed 41.33% ± 3.43 mortality due to V. microstoma infestation. However, none of the ciliates were effective against Ae. aegypti larvae. Chilodonella sp. was very occasionally reported during this study hence was not possible to the mass rear for experimentations. This study concludes that V. microstoma is an effective ciliated parasite of Cx. tritaeniorhynchus larvae. Due to their effectiveness and eco-friendly nature, this species can be developed as an effective bio-controlling agent against Cx. tritaeniorhynchus mosquito species.

Diversity of mosquito species and potential arbovirus transmission in long-tailed macaque (Macaca fascicularis) breeding facilities

Background and Aim:Mosquito-borne viral infections are diseases that reduce human and animal health levels. Their transmission involves wildlife animals as reservoirs and amplifying hosts, including long-tailed macaques (Macaca fascicularis), and potentially transmits to humans and vice versa. This study aimed to determine the species diversity, richness, and biting activity of mosquitoes in a long-tailed macaque breeding area facility and discover the presence of Flavivirus and Alphavirus as the two main arboviruses reported to infect macaques.Materials and Methods:Human landing catch, light trap, and sweep net methods were used for mosquito collection around long-tailed macaques cages at parallel times for 12 h (18:00–06:00) for 12 nights. Mosquito species were identified to the species level based on the morphological identification key for Indonesian mosquitoes. Mosquito diversity was analyzed by several diversity indices. Mosquitoes caught using the human landing catch method were pooled based on mosquito species for viral ribonucleic acid extraction. Reverse transcription-polymerase chain reaction (RT-PCR) detected the non-structural protein 5 of the Flavivirus region and the non-structural protein 4 of the Alphavirus region. This study used the man-hour density and man-biting rate formulas for mosquito density.Results:Ten mosquito species were collected, namely, Aedes albopictus, Anopheles aconitus, Anopheles minimus, Anopheles vagus, Armigeres foliatus, Armigeres subalbatus, Culex gelidus, Culex hutchinsoni, Culex tritaeniorhynchus, and Culex quinquefasciatus. The number of mosquitoes caught using the light trap method had the highest abundance. In contrast, the number of mosquito species caught using the sweep net method had lower diversity than the other two methods. Seven mosquito species were obtained using the human landing catch method. The mosquito species with the highest density was Cx. quinquefasciatus within the observed densest period from 20:00 to 21:00. Negative results were obtained from RT-PCR testing on five species detected using universal Flavivirus and Alphavirus primers.Conclusion:The occurrence of mosquitoes in long-tailed macaque breeding facilities can be a source of transmission of zoonotic vector-borne diseases between animals and humans and vice versa.

Molecular characterization and genetic divergence of seven Culex mosquito (Diptera: Culicidae) species using Mt COI gene from Odisha State, India

BackgroundCulex mosquitoes are involved in the transmission of arboviral diseases worldwide. Bio-ecology and identification of mosquitoes are of paramount importance to develop species-specific vector control strategies. Genetic-based species-specific approaches that reduce the burden of vector-borne diseases are made possible by molecular techniques.ResultsIn the present study, the mitochondrial COI gene of Culex mosquitoes was used for molecular identification in addition to morpho-taxonomy. Our findings suggested the presence of important Culex mosquito vectors viz, Culex vishnui, Culex tritaeniorhynchus, Culex bitaeniorhynchus, Culex quinquefasciatus, Culex gelidus, Culex fuscocephala, and Culex fuscanus in the southern part of Odisha state, India. We examined the phylogeny and genetic diversity of the above seven different Culex populations from different geographical locations. An average intra-specific K2P distance of the COI gene was found to be 0.9%. Further, to measure the diversity of the Culex population among different geographical strains, haplotype diversity and nucleotide diversity were compared. Culex fuscanus showed high polymorphisms and mutations with high nucleotide diversity (0.013) and the Culex quinquefasciatus showed the lowest variation in P(i), 0.0013 in the intra-population polymorphism analysis of COI sequences. Similarly, the Haplotype diversity (Hd) found in Culex gelidus and Culex fuscocephala with the value of 0.972 and Culex quinquefasciatus (0.583) showed the lowest value of haplotype diversity. A haplotype network was constructed to establish the genealogical relationship between haplotypes. The phylogenetic tree was constructed that produces distinctive conspecific clusters in different Culex species. Population genetic study has illustrated the occurrence of genetic differentiation within the population.ConclusionsThe findings of this study contribute to greater evidence that DNA barcode sequences can be used to monitor mosquito species diversity. This study also adds valuable information about the systematics and molecular biology of seven public health important mosquito species acting as a significant vector for Japanese encephalitis in various Asian continents. This information is further used for the effective implementation of region-specific vector control strategies.

UV Light-Emitting-Diode Traps for Collecting Nocturnal Biting Mosquitoes in Urban Bangkok

Simple SummaryThis study was conducted to evaluate the efficacy of six ultraviolet light-emitting diodes (UV-LED) traps and a fluorescent light trap for sampling urban nocturnal mosquitoes. Results demonstrated that the fluorescent light trap outperformed all the UV-LED traps throughout the 72 sampling nights and between wet and dry seasons. Among the UV-LED traps, the LED375 trapped the highest number of mosquitoes. Additional field trials are needed to validate these findings in different ecological settings.Well-designed surveillance systems are required to facilitate a control program for vector-borne diseases. Light traps have long been used to sample large numbers of insect species and are regarded as one of the standard choices for baseline insect surveys. The objective of this study was to evaluate the efficacy of six ultraviolet light-emitting diodes and one fluorescent light for trapping urban nocturnal mosquito species within the Kasetsart University (KU), Bangkok. Ultraviolet light-emitting diodes (UV-LEDs), (LED365, LED375, LED385, LED395, and LED405) and a fluorescent light were randomly assigned to six different locations around the campus in a Latin square design. The traps were operated continuously from 18:00 h to 06:00 h throughout the night. The traps were rotated between six locations for 72 collection-nights during the dry and wet seasons. In total, 6929 adult mosquitoes were caught, with the most predominant genus being Culex, followed by Aedes, Anopheles, Armigeres and Mansonia. Among the Culex species, Culex quinquefasciatus (n = 5121: 73.9%) was the most abundant followed by Culex gelidus (n = 1134: 16.4%) and Culex vishnui (n = 21: 0.3%). Small numbers of Aedes, Armigeres, and Anopheles mosquitoes were trapped [Aedes albopictus (n = 219: 3.2%), Aedes pocilius (n = 137: 2.0%), Armigeres subalbatus (n = 97: 1.4%), Anopheles vagus (n = 70: 1.0%), Aedes aegypti (n = 23: 0.3%)]. There were 2582 specimens (37.2%) captured in fluorescent light traps, whereas 942 (13.6%), 934 (13.5%), 854 (12.3%), 820 (11.8%), and 797 (11.5%) were captured in the LED375, LED405, LED395, LED365, and LED385 traps, respectively. None of the UV-LED light traps were as efficacious for sampling nocturnal mosquito species as the fluorescent light trap. Among the five UV-LED light sources, LED375 trapped the greatest number of mosquitoes. Additional field trials are needed to validate these findings in different settings in order to substantially assess the potential of the LEDs to trap outdoor nocturnal mosquitoes.

The Emergence of Japanese Encephalitis Virus in Australia in 2022: Existing Knowledge of Mosquito Vectors.

In early 2022, the Japanese encephalitis virus (JEV) was identified as the cause of stillborn and mummified piglets in pig farms in southeastern Australia. Human cases and additional pig farms with infected piglets were subsequently identified across a widespread area encompassing four states. To inform surveillance and control programs, we synthesized existing information on Australian vectors of JEV, much of which was generated in response to incursions of JEV into the northern state of Queensland between 1995 and 2005. Members of the Culex sitiens subgroup, particularly Culex annulirostris, should be considered the primary vectors of JEV in Australia, as they yielded >87% of field detections of JEV, were highly efficient laboratory vectors of the virus, readily fed on pigs and birds (the key amplifying hosts of the virus) when they were available, and are widespread and often occur in large populations. Three introduced species, Culex quinquefasciatus, Culex gelidus and Culex tritaeniorhynchus may also serve as vectors, but more information on their geographical distribution, abundance and bionomics in the Australian context is required. Mosquitoes from other genera, such as Aedes and Verrallina, whilst considered relatively poor vectors, could play a regional or supplemental role in transmission, especially facilitating vertical transmission as a virus overwintering mechanism. Additional factors that could impact JEV transmission, including mosquito survival, dispersal and genetics, are also discussed. Possible directions for investigation are provided, especially in the context of the virus emerging in a region with different mosquito fauna and environmental drivers than northern Australia.

Composition of Mosquito Species in Three Selected Villages in Mahabubnagar District, Telangana

Mosquito prevalence was studied in three areas of Mahabubnagar district of Telangana viz. Mannanur,Janampeta, B Veerapur. Mosquito sampling was done in three selected villages in Mahabubnagar districtin all seasons from 2014-2016. Density, Distribution and habitat preferences are studied for three years2014- 2016. A total of 1787 mosquitoes were collected. Identification was done using identification keysof Christophers (1933) and Barraud (1934). Three genera Culex, Aedes and Anopheles were observed.Anopheles contributes 44%, Culex 39% and Aedes contributes 17%. Culex quinquefasciatus is predominantspecies (16%), while Aedes albopictus (15%) and Culex gelidus (15%) occupy the second place. Anophelesculicifacies contributes 12%, while Culex tritaeniorhynchus (8%) and Anopheles vagus (8%) contributeequally. Anopheles subpictus (7%), and Anopheles stephensi (7%) contributed equally, while Aedes aegyptiis with only 5%. Human dwellings inhabit 50% due to breeding sites; cattle shed have 25%, and rainwater ditch have very less, as these are available only in monsoons. Cx. quinquefasciatus, Cx. gelidus werefound in all habitats, while An. subpictus, An. stephensi, An. culicifacies, Ae. aegypti, Ae. albopictus werefound in some of the habitats.

Japanese encephalitis virus genotype III from mosquitoes in Tarlac, Philippines

ObjectivesThe aim of this study was to investigate the presence of Japanese encephalitis virus (JEV) in a rice-farming community in the Philippines and to determine its implications regarding the epidemiology of viral encephalitides in the Asia-Pacific Region. MethodsMosquitoes were collected monthly from animal-baited traps close to flooded rice fields in two barangays (villages) in the Municipality of San Jose, Tarlac Province in Luzon, from May 2009 to July 2010. Virus was detected by nested reverse transcription PCR. Phylogenetic analysis of the amplified virus envelope gene was done using the maximum-likelihood method. ResultsA total of 28 700 known vector mosquitoes were collected, namely Culex vishnui, Culex fuscocephala, Culex tritaeniorhynchus, and Culex gelidus. JEV genotype III was detected in C. tritaeniorhynchus, belonging to the same genotype but form a different clade from those reported in the 1980s and in 2020 in this country. ConclusionsJapanese encephalitis is associated with rice cultivation and the presence of infected mosquitoes in Tarlac, Philippines. It remains to be seen whether the observed genetic shift of genotype III to genotype I in Asia will in time have an impact on the epidemiology of Japanese encephalitis in the Philippines. For long-term disease control, regular surveillance and Japanese encephalitis immunization in children and travelers in high risk areas are recommended.

The abundance of mosquito larvae in relation to associated microbiota and physicochemical parameters of marshland habitats in Kelaniya in Gampaha district of Sri Lanka

The abundance of mosquito larvae in relation to associated microbiota and physicochemical parameters of water in marshes from selected habitats in Kelaniya divisional secretariat area, of the Gampaha district, Sri Lanka were studied. Culex gelidus was found to be the dominant mosquito species found in marshes followed by Culex whitmorei, Culex tritaeniorhynchus, Culex quinquefasciatus, Culex fuscocephala, and Anopheles peytoni. Fourteen species/taxa of microbiota were identified. There was a significant positive correlation between the abundance of Cx. gelidus larvae and level of organic pollution in marshes as measured by the five-day Biological Oxygen Demand. A positive correlation for the presence of Cx. fuscocephala and Cx. gelidus in the marshland was also observed. Nevertheless, abundance of Cx. gelidus was negatively correlated with Cx. quinquefasciatus and Cx. whitmorei. The presence of Culex gelidus and Cx. tritaeniorhynchus were positively correlated with Daphnia magna indicating their co-existence in the same habitat requirement. Culex gelidus was negatively affected by the epibiont, Vorticella microstoma, when the latter organisms were present in higher densities. In contrast, the presence of Cx. whitmorei was positively influenced by the Vorticella microstoma, Difflugia corona, nauplius larva, and Keratella valga. There was a negative correlation between Cx. whitmorei and Monostyla bulla (Rotifera; Monogononta) and between the abundance of Cx. fuscocephala and Lecane luna in the same marshland. This study concludes that mosquito larvae and some microbiota are interdependent in marshlands. The level of abundance of mosquito larvae and microbiota varied with the level of water pollution. Cx. gelidus larvae are biologically affected by Vorticella microstoma. Many microbiota species compete with mosquito larvae for the same food items.

Culex gelidus

This datasheet on Culex gelidus covers Identity, Distribution, Hosts/Species Affected.

Larval Ecology of Anopheles Mosquitoes in Kudat, Sabah

The emergence of human Plasmodium knowlesi malaria appeared to have been precipitated by the displacement of the natural environment of macaques and Anopheles mosquitoes resulting from deforestation and land-use changes in Malaysia. A longitudinal survey of larval habitats was conducted from May 2015 to April 2016 in the District of Kudat, Sabah to better understand how these changes have affected mosquitoes across six land use categories. Larvae were collected by dipping and reared in the laboratory for the identification of adults. Five anopheline and three culicine species were present: Anopheles balabacensis, An. barbirostris, An. lesteri, An. borneensis, An. umbrosus, Aedes albopictus, Culex gelidus, and Toxorhynchites sp. An. balabacensis was found in all six land-use types. Biodiversity by genera was high in all land-use types. The relative importance of land use types and larval habitats as sources of potential vectors was analyzed by the Kruskal-Wallis H test by ranks. In decreasing order Anopheles larvae were found in rubber tree plantation &gt; coconut plantation &gt; clearing site &gt; palm oil plantation &gt; forest &gt; settlement area. Important larval habitats were intermittent stream &gt; ditch &gt; pond &gt; artificial container &gt; puddle &gt; river &gt; slow-flowing stream. Eighteen breeding sites of An. balabacensis were within (500 m) the average maximum flight range of the species and houses at risk for malaria. Knowledge gained from the study can be used to assess the need for vector control in preventing the spread of P. knowlesi in vulnerable areas.

Identification and seasonal distribution of mosquito species in Telangana state, India

An entomological study was conducted to determine species composition, relative abundance, and seasonal population fluctuation of mosquitoes. Thirty localities were randomly selected and sampled once in summer, monsoon and winter seasons for a period of three years. Adult mosquitoes were collected from in and around human dwellings and animal sheds using aspirator and flash light between 6 PM and 7:30 PM hrs and 06:00 AM and 7:00 AM hrs. Larvae were collected from ditches, rock pool and other domestic habitats, reared to formation of adults in the lab. A total of 18425 mosquitoes were collected. Five genera (Culex, Anopheles, Aedes, Mansonia and Armigeres) were found to be circulating. Further, with respect to habitat, more number of mosquitoes were observed around cattle sheds and houses. Culex quinquefasciatus occupies top place, followed by Aedes albopictus and Culex gelidus, Culex tritaeniorhynchus with respect to density, whereas Mansonia uniformis occupies last place. Culex quinquifasciatus is top again followed by Culex gelidus and Aedes albopictus, Mansonia uniformis is last again with respect to distribution. The vector species studied were Aedes aegypti, Aedes albopictus for chikungunya and dengue, Anopheles stephensi, Anopheles culicifacies for malaria, Culex tritaeniorhynchus for Japanese encephalitis and Culex quiquefasciatus for lymphatic filaria.

Comparative Analysis of the Gut Microbiota of Adult Mosquitoes From Eight Locations in Hainan, China.

The midgut microbial community composition, structure, and function of field-collected mosquitoes may provide a way to exploit microbial function for mosquito-borne disease control. However, it is unclear how adult mosquitoes acquire their microbiome, how the microbiome affects life history traits and how the microbiome influences community structure. We analyzed the composition of 501 midgut bacterial communities from field-collected adult female mosquitoes, including Aedes albopictus, Aedes galloisi, Culex pallidothorax, Culex pipiens, Culex gelidus, and Armigeres subalbatus, across eight habitats using the HiSeq 4000 system and the V3−V4 hyper-variable region of 16S rRNA gene. After quality filtering and rarefaction, a total of 1421 operational taxonomic units, belonging to 29 phyla, 44 families, and 43 genera were identified. Proteobacteria (75.67%) were the most common phylum, followed by Firmicutes (10.38%), Bacteroidetes (6.87%), Thermi (4.60%), and Actinobacteria (1.58%). The genera Rickettsiaceae (33.00%), Enterobacteriaceae (20.27%), Enterococcaceae (7.49%), Aeromonadaceae (7.00%), Thermaceae (4.52%), and Moraxellaceae (4.31%) were dominant in the samples analyzed and accounted for 76.59% of the total genera. We characterized the midgut bacterial communities of six mosquito species in Hainan province, China. The gut bacterial communities were different in composition and abundance, among locations, for all mosquito species. There were significant differences in the gut microbial composition between some species and substantial variation in the gut microbiota between individuals of the same mosquito species. There was a marked variation in different mosquito gut microbiota within the same location. These results might be useful in the identification of microbial communities that could be exploited for disease control.

Mosquito Diversity and Population Genetic Structure of Six Mosquito Species From Hainan Island.

Hainan is a tropical island in southern China with abundant mosquito species, putting Hainan at risk of mosquito-borne virus disease outbreaks. The population genetic diversity of most mosquito species on Hainan Island remains elusive. In this study, we report the diversity of mosquito species and the genetic diversity of the predominant species on Hainan. Field populations of adults or larvae were collected from 12 regions of Hainan Island in 2018 and 2019. A fragment of the mitochondrial cytochrome c oxidase subunit I (coxI) gene was sequenced from 1,228 mosquito samples and used for species identification and genetic diversity analysis. Twenty-three known mosquito species from the genera Aedes, Armigeres, Culex, Mansonia, and Anopheles and nine unconfirmed mosquito species were identified. Aedes albopictus, Armigeres subalbatus, and Culex pipiens quinquefasciatus were the most prevalent mosquito species on Hainan. The regions north of Danzhou, Tunchang, and Qionghai exhibited high mosquito diversity (26 species). The order of the total haplotype diversity and nucleotide diversity of the populations from high to low was as follows: Culex tritaeniorhynchus, Ar. subalbatus, Culex pallidothorax, Culex gelidus, Ae. albopictus, and C. p. quinquefasciatus. Tajima’s D and Fu’s Fs tests showed that Ae. albopictus, C. p. quinquefasciatus, C. tritaeniorhynchus, and C. gelidus had experienced population expansion, while the Ar. subalbatus and C. pallidothorax populations were in genetic equilibrium. Significant genetic differentiation existed in the overall populations of Ae. albopictus, Ar. subalbatus, C. p. quinquefasciatus, and C. pallidothorax. The Ae. albopictus populations on Hainan were characterized by frequent gene exchange with populations from Guangdong and four other tropical countries, raising the risk of viral disease outbreaks in these regions. Two subgroups were reported in the Ar. subalbatus populations for the first time. Our findings may have important implications for vector control on Hainan Island.

Molecular Characterization of Culturable Aerobic Bacteria in the Midgut of Field-Caught Culex tritaeniorhynchus, Culex gelidus, and Mansonia annulifera Mosquitoes in the Gampaha District of Sri Lanka.

Background Larval and adult mosquito stages harbor different extracellular microbes exhibiting various functions in their digestive tract including host-parasite interactions. Midgut symbiotic bacteria can be genetically exploited to express molecules within the vectors, altering vector competency and potential for disease transmission. Therefore, identification of mosquito gut inhabiting microbiota is of ample importance before developing novel vector control strategies that involve modification of vectors. Method Adult mosquitoes of Culex tritaeniorhynchus, Culex gelidus, and Mansonia annulifera were collected from selected Medical Officer of Health (MOH) areas in the Gampaha district of Sri Lanka. Midgut lysates of the field-caught non-blood-fed female mosquitoes were cultured in Plate Count Agar medium, and Prokaryotic 16S ribosomal RNA partial genes of the isolated bacteria colonies were amplified followed by DNA sequencing. Diversity indices were used to assess the diversity and richness of the bacterial isolates in three mosquito species. The distribution pattern of bacterial isolates between different mosquito species was assessed by Distance-Based Redundancy Analysis (dbRDA). Results A total of 20 bacterial species (Staphylococcus pasteuri, Bacillus megaterium, Staphylococcus cohnii, Pantoea dispersa, Staphylococcus chromogenes, Bacillus aquimaris, Staphylococcus arlettae, Staphylococcus sciuri, Staphylococcus warneri, Moraxella osloensis, Enterobacter sp., Klebsiella michiganensis, Staphylococcus hominis, Staphylococcus saprophyticus, Streptomyces sp., Bacillus niacin, Cedecea neteri, Micrococcus luteus, Lysinibacillus sphaericus, and Bacillus licheniformis) were identified. All of these species belonged to three phyla, Proteobacteria, Firmicutes, and Actinobacteria, out of which phylum Firmicutes (71.1%) was the most prominent. The least number of species was recorded from Actinobacteria. The relative distribution of midgut microbes in different mosquito species differed significantly among mosquito species (Chi-square, χ2 = 486.091; df = 36; P ≤ 0.001). Midgut microbiota of Cx. tritaeniorhynchus and Cx. gelidus indicated a similarity of 21.51%, while Ma. annulifera shared a similarity of 6.92% with the cluster of above two species. The gut microbiota of Cx. tritaeniorhynchus was also significantly more diverse and more evenly distributed compared to Ma. annulifera. Simpson's diversity, Margalef's diversity, and Menhinick's diversity indices were higher in Cx. gelidus. Of the recorded species, P. dispersa and strains of nonpathogenic species in Bacillaceae family (B. megaterium, B. niacini, B. licheniformis, and L. sphaericus) can be recommended as potential candidates for paratransgenesis. Conclusion The relative distribution of midgut microbes in different mosquito species differed significantly among the three studied adult mosquito species. The present data strongly encourage further investigations to explore the potential usage of these microbes through paratransgenic approach for novel eco-friendly vector control strategies.

Entomological investigations on sporadic Japanese encephalitis sero-positivity in Tamil Nadu, India

In India, Japanese Encephalitis (JE) continues to be a public health issue in some parts of our country. JE surveillance includes early reporting of clinical cases, sentinel sero-surveys and vector surveillance in the endemic areas. In the present study, JE longitudinal vector surveillance and epidemiological investigations were carried out for the first time during two consecutive years in the endemic district of Tamil Nadu. 22,538 mosquitoes were collected, species identified and screened for JE virus by RT-PCR. Predominant was Culex tritaeniorhynchus (60%) and followed by it Anopheles subpictus (23%), Culex quinquefasciatus (8%) and Culex gelidus (3%). It suggests that Culex tritaeniorhynchus may act as major vector and An. subpictus may act as secondary vector. Monsoon and post-monsoon seasons favour breeding of Cx. tritaeniorhynchus leading to vector abundance. Preferential resting sites for Cx. tritaeniorhynchus were pig and cattle shed. Although clinical cases have been reported seasonally in the three blocks, the presence of virus among field caught mosquitoes could not be established by RT-PCR. It may be due to the low titre value of JE virus in mosquitoes. This is the first report of JE investigations in the endemic district of Tamil Nadu and it helps to formulate the effective control strategies for JE virus transmission.