Mosquito Research Articles

The First Isolation of Insect-Specific Alphavirus (Agua Salud alphavirus) in Culex (Melanoconion) Mosquitoes in the Brazilian Amazon.

Advances in diagnostic techniques coupled with ongoing environmental changes have resulted in intensified surveillance and monitoring of arbovirus circulation in the Amazon. This increased effort has resulted in increased detection of insect-specific viruses among hematophagous arthropods collected in the field. This study aimed to document the first isolation of Agua Salud alphavirus in mosquitoes collected within the Brazilian Amazon. Arthropods belonging to the family Culicidae were collected within a forest fragment located in the Environmental Protection Area of the metropolitan region of Belem. Subsequently, these specimens were meticulously identified to the species level. Afterward, the collected batches were macerated, and the resulting supernatant was then inoculated into C6/36 and Vero cell cultures to facilitate viral isolation. The presence of arboviruses within the inoculated cell cultures was determined through indirect immunofluorescence analysis. Furthermore, positive supernatant samples underwent nucleotide sequencing to precisely identify the viral strains present. Notably, a batch containing Culex (Melanoconion) mosquitoes was identified to be positive for the genus Alphavirus via indirect immunofluorescence. This study is the first report on insect-specific alphavirus isolation in Brazil and the first-ever description of Agua Salud alphavirus isolation within Amazon Forest remnants.

New Iflavirus Species Characterized from Mosquitoes Captured in the Sao Paulo Zoological Facilities.

Metagenomic studies of mosquito viromes demonstrated a more diverse composition than just an exclusive composition of pathogenic arboviruses transmitted to humans. In our study, the virome of 866 female mosquitoes collected throughout 2020 at the São Paulo Zoo, located in the city of São Paulo/SP-Brazil, was obtained. Specifically, in this paper, we describe a new virus found by viral RNA extraction and next-generation MiSeq sequencing of a group of 23 specimens of Anopheles (Nys.) strodei. The complete genome with a length of 9709 nucleotides was characterized by a positive orientation and a single strand, with a single large ORF, which encodes a polyprotein of 2987 amino acids. The phylogenetic analysis showed an association with the viral family Iflaviridae and the Riboviria realm. We carried out comparisons with translated sequences of the capsid regions of other iflavirus, and the identities in relation to our sequence were below the minimum limit of 90%, indicating that possibly it is a new species of iflavirus. Our findings contribute to expanding knowledge of virome composition among mosquito species in Brazil and globally. Moreover, we provide a viral genome reference specific to this geographic region and Culicidae family of mosquitoes. This resource facilitates future in silico recognition and assembly of viral genomes within metagenomic datasets.

First detection of Rift Valley Fever Virus among Culex pipiens in Tahoua, Niger

The Rift Valley Fever (RVF) is an arbovirus disease responsible of regular epizootics and epidemics in sub-Saharan Africa and Arabian Peninsula. In 2016, Niger experienced its first outbreak of RVF in Tahoua region, which resulted in high consequences in animal and human health. The aim of this study was to investigate on the RVFV circulation among potential vectors of the disease. This was a cross-sectional survey carried out in Tahoua and Agadez regions in August 2021. Adult mosquitoes were collected by using the morning spray in human dwellings and the CDC light trap methods. After morphological identification, viral RNA was extracted. The RNA was extracted by using QIAamp Viral RNA Mini Kit (Qiagen). The RVFV detection was performed by using the qRT-PCR method. A total of 2487 insects (1978 mosquitoes, 509 sandflies and 251 biting midges) were identified and divided into three families (Culicidae, Psychodidae and Ceratopogonidae). The Culicidae family composed of the Culex genus being the most abundant with a predominance of Cx.pipiens (31.88%; n = 793) followed by Mansonia sp (21.51%; n = 535), Anophelesgambiae s.l. (8.44%; n = 210), An. pharoensis (0.72%; n = 18), An. rufipes (0.48%; n = 12), Cx. quinquefasciatus (6.39%; n = 159), the Psychodidae with sandflies (20.46%; n = 509), and the Ceratopogonidae with Culicoides genus (10.09%; n = 251). The qRT-PCR carried out on a sample of mosquitoes (N = 96) highlighted that one individual of Cx.pipiens was found positive to RVFV. This specimen was from Tassara locality (Tahoua) and collected by CDC Light Trap method. This study reveals for the first time the circulation of RVFV among Cx.pipiens in Niger and highlights the possible vectorial role of this vector in the disease transmission. Further investigations should be carried out to identify the biological and ecological determinants that support the maintenance of the virus in this area in order to guide control interventions.

Human dirofilariasis: a rare disease or a problem of our time?

The article presents the current state of the problem of dirofilariasis. Helminth infection caused by nematodes of the genus Dirofilaria is mainly a disease of carnivores from the canine and feline families. Humans are considered occasional hosts in which Dirofilaria spp., more commonly D. repens, rarely reach sexual maturity. The climatic conditions of the territory are one of the determining factors for the spread of vector-borne diseases. Due to the fact that in recent years there has been a tendency to increase average temperatures everywhere, these changes lead to a shift and expansion of the ranges of many organisms, including pathogens and vectors of various diseases. The clinical picture of human dirofilariasis caused by D. repens most often manifests itself in one of two clinical forms -subcutaneous and ocular, although, according to the literature, there are cases of damage to other localizations. Due to the fact that damage to the organs of vision occurs most often in humans, the clinical case presented by us is relevant. The patient, in our clinical case, permanently lived in one of the districts of the Ryazan region. The landscape and natural complex of this region, taking into account the presence of a large number of wetlands, an abundance of water bodies and suitable climatic conditions, creates a favorable environment for the development of insects of the Culicidae family (mosquitoes of the genera Anopheles, Culex, etc.), which are transmissible carriers of various human and animal diseases, including dirofilariasis. The complex cycle of the parasite, the possibility of migration in the body can lead to multiple organ damage. Scientific publications describe cases of finding dirofilaria in the lungs, heart, and genitourinary system. In this regard, the awareness of doctors and patients about this disease is very relevant, and will reduce the likelihood of prolonging the correct diagnosis and treatment.

Isolation and molecular detection of dengue and chikungunya virus from field-collected adult mosquitoes in Kelantan, Malaysia.

Dengue and chikungunya infections are one of the major health problems that have plagued the human population globally. All dengue virus (DENV) serotypes circulate within Malaysia with particular serotypes dominating in different years/outbreaks. In the state of Kelantan, an increasing number of DENV and chikungunya virus (CHIKV) new cases have been reported, including several deaths. This study aimed to isolate and detect these arboviruses from adult mosquitoes in Kelantan. Adult mo squito samples were collected from January to August 2019 and were identified according to gender, species and locality. The isolation of the virus was done in C6/36 cells. Dengue NS1 antigen was carried out using direct mosquito lysate and mosquito culture supernatant. Detection and serotyping of the DENV was performed using multiplex RT-PCR and CHIKV detection using a one-step RT-PCR assay. Of 91 mosquito pools, four were positive for NS1 antigen comprising two pools (2.2%) of male Ae. albopictus (Pulau Melaka and Kubang Siput) and two pools (2.2%) of Ae. aegypti (Kampung Demit Sungai). DENV 1 was detected in one pool (0.9%) of female Ae. albopictus among 114 tested Aedes pools. Two pools of 114 pools (1.7%) from both male Aedes species were positive with double serotypes, DENV 1 and DENV 2 (Pulau Melaka). However, no pool was positive for CHIKV. The presence of DENV and the main vectors of arboviruses in Kelantan are pertinent indicators of the need to improve vector controls to reduce arbovirus infections among people in the localities.

On the Origin and Evolution of the Mosquito Male-determining Factor Nix.

The mosquito family Culicidae is divided into 2 subfamilies named the Culicinae and Anophelinae. Nix, the dominant male-determining factor, has only been found in the culicines Aedes aegypti and Aedes albopictus, 2 important arboviral vectors that belong to the subgenus Stegomyia. Here we performed sex-specific whole-genome sequencing and RNAseq of divergent mosquito species and explored additional male-inclusive datasets to investigate the distribution of Nix. Except for the Culex genus, Nix homologs were found in all species surveyed from the Culicinae subfamily, including 12 additional species from 3 highly divergent tribes comprising 4 genera, suggesting Nix originated at least 133 to 165 million years ago (MYA). Heterologous expression of 1 of 3 divergent Nix open reading frames (ORFs) in Ae. aegypti resulted in partial masculinization of genetic females as evidenced by morphology and doublesex splicing. Phylogenetic analysis suggests Nix is related to femaleless (fle), a recently described intermediate sex-determining factor found exclusively in anopheline mosquitoes. Nix from all species has a conserved structure, including 3 RNA-recognition motifs (RRMs), as does fle. However, Nix has evolved at a much faster rate than fle. The RRM3 of both Nix and fle are distantly related to the single RRM of a widely distributed and conserved splicing factor transformer-2 (tra2). The RRM3-based phylogenetic analysis suggests this domain in Nix and fle may have evolved from tra2 or a tra2-related gene in a common ancestor of mosquitoes. Our results provide insights into the evolution of sex determination in mosquitoes and will inform broad applications of mosquito-control strategies based on manipulating sex ratios toward nonbiting males.

Characterizing viral species in mosquitoes (Culicidae) in the Colombian Orinoco: insights from a preliminary metagenomic study

Mosquitoes (Diptera: Culicidae) are primary vectors of arthropod-borne viruses (arboviruses) that pose significant public health threats. Recent advances in sequencing technology emphasize the importance of understanding the arboviruses and insect-specific viruses (ISVs) hosted by mosquitoes, collectively called the “virome”. Colombia, a tropical country with favorable conditions for the development and adaptation of multiple species of Culicidae, offers a favorable scenario for the transmission of epidemiologically important arboviruses. However, entomovirological surveillance studies are scarce in rural areas of the country, where humans, mosquitoes, and animals (both domestic and wild) coexist, leading to a higher risk of transmission of zoonotic diseases to humans. Thus, our study aimed to perform a preliminary metagenomic analysis of the mosquitoes of special relevance to public health belonging to the genera Ochlerotatus, Culex, Limatus, Mansonia, Psorophora, and Sabethes, within a rural savanna ecosystem in the Colombian Orinoco. We employed third-generation sequencing technology (Oxford Nanopore Technologies; ONT) to describe the virome of mosquitoes samples. Our results revealed that the virome was primarily shaped by insect-specific viruses (ISVs), with the Iflaviridae family being the most prevalent across all mosquito samples. Furthermore, we identified a group of ISVs that were common in all mosquito species tested, displaying the highest relative abundance concerning other groups of viruses. Notably, Hanko iflavirus-1 was especially prevalent in Culex eknomios (88.4%) and Ochlerotatus serratus (88.0%). Additionally, other ISVs, such as Guadalupe mosquito virus (GMV), Hubei mosquito virus1 (HMV1), Uxmal virus, Tanay virus, Cordoba virus, and Castlerea virus (all belonging to the Negevirus genus), were found as common viral species among the mosquitoes, although in lower proportions. These initial findings contribute to our understanding of ISVs within mosquito vectors of the Culicidae family in the Eastern Plains of Colombia. We recommend that future research explore deeper into ISV species shared among diverse vector species, and their potential interactions with arboviruses. In addition, we also showed the need for a thorough exploration of the influence of local rural habitat conditions on the shape of the virome in mosquito vectors.

Catalase enzymatic activity in adult mosquitoes (Diptera: Culicidae): taxonomic distribution of the continuous trait suggests its relevance for phylogeny research.

Molecular research based on gene sequence analysis and performed for decades, in general, supported morphology-based groupings of the species within the family Culicidae, but phylogenetic relationships between some genera and tribes remained uncertain for a long time. Interspecific differences in catalase, an antioxidant enzyme important for maintaining prolonged lifespan and reproduction, have not been studied extensively by estimating enzymatic activity levels. Here, catalase enzymatic activity was assayed in extracts of male mosquitoes belonging to 10 species of the subfamily Culicinae, including species from tribes of disputable phylogeny. Three species of Chaoboridae (nearest outgroup taxon) and mosquitoes from the subfamily Anophelinae (one species complex) were also added to the study. At least in Culicinae, immature adult males (less than one day after emergence) have distinctly elevated specific activity of catalase; therefore, only mature males of all species were used for the comparative study. As a result, significant differences in catalase activity were revealed between tribes, genera and particular species. Among culicids, the genera Coquillettidia and Culiseta were found to include the species with the highest and relatively high catalase activity, which is consistent with the affinity of the tribes Mansoniini and Culisetini to each other. Within Ochlerotatus, extremely low catalase activity in Oc. hexodontus suggests the more distant position of this species from Oc. cantans (Meigen) and Oc. communis (de Geer) than the positions of the latter two species from each other. Additional study of catalase activity in overwintering females of the genus Culex revealed significantly higher enzyme activity in Cx. torrentium in comparison with Cx. pipiens, which supports their quite distant positions from each other within the genus. Considering the distribution of catalase activity within the tree obtained, the preliminary outcome is that Culiseta retains the elevated level of catalase activity that was lost during the early separation of Anopheles and subsequent separation of Culex and Aedes/Ochlerotatus after Anopheles from their common branch with Culiseta/Coquillettidia. Overall, the use of taxonomic distribution of catalase activity levels appears to be effective for resolving disputed events of mosquito phylogeny.

Mitogenome-based phylogeny of mosquitoes (Diptera: Culicidae).

Mosquitoes are of great medical significance as vectors of many deadly diseases. Mitogenomes have been widely used in phylogenetic studies, but mitogenome knowledge within the family Culicidae is limited, and Culicidae phylogeny is far from resolved. In this study, we surveyed the mitogenomes of 149 Culicidae species, including 7 newly sequenced species. Comparative analysis of 149 mosquito mitogenomes shows gene composition and order to be identical to that of an ancestral insect, and the AT bias, length variation, and codon usage are all consistent with that of other reported Dipteran mitogenomes. Phylogenetic analyses based on the DNA sequences of the 13 protein-coding genes from the 149 species robustly support the monophyly of the subfamily Anophelinae and the tribes Aedini, Culicini, Mansoniini, Sabethini, and Toxorhynchitini. To resolve ambiguous relationships between clades within the subfamily Culicinae, we performed topological tests and show that Aedini is a sister to Culicini and that Uranotaeniini is a sister to (Mansoniini+(Toxorhynchitini+Sabethini)). In addition, we estimated divergence times using a Bayesian relaxation clock based on the sequence data and 3 fossil calibration points. The results show mosquitoes diverged during the Early Jurassic with massive Culicinae radiations during the Cretaceous, coincident with the emergence of angiosperms and the burst of mammals and birds. Overall, this study, which uses the largest number of Culicidae mitogenomes sequenced to date, comprehensively reveals the mitogenome characteristics and mitogenome-based phylogeny and divergence times of Culicidae, providing information for further studies on the mitogenome, phylogeny, evolution, and taxonomic revision of Culicidae.

A cross-sectional study of malaria transmission in suggests the existence of a potential bridge vector susceptible of ensuring the transfer of simian malaria parasites to humans

IntroductionDespite all the efforts made to control or even eliminate malaria, the disease continues to claim the highest number of victims of vector-borne pathogens in the world and Sub-Saharan countries bear the heaviest burden. The lack of knowledge of the role of various protagonists involved in the transmission of this parasitic disease, such as mosquito vectors and the plasmodial species they transmit as well as the host species they infect in a locality, constitutes one of the main causes of the persistence of malaria. In Gabon, in several areas, entomological data on malaria transmission remain poorly known. Thus, this study aimed to determine the diversity of Anopheles involved in malaria transmission in different environments of the province of Nyanga in southwest Gabon.MethodsFor this, an entomological study was carried out in the four main localities of the province of Nyanga to provide answers to these shortcomings. Mosquitoes were collected over several nights using the human landing catch method. The identification of Anopheles and malaria parasites circulating in the different sites was achieved by combining morphological and molecular analysis tools.ResultsA total of five hundred and ninety-one (591) mosquitoes belonging to the Culicidae family were collected. From this collection of adult mosquitoes, nine species of Anopheles mosquitoes notably species of the Anopheles nili complex (53.46%) followed by those of Anopheles gambiae complex (22.01%), Anopheles funestus group (18.24%), Anopheles moucheti complex (5.66%) and Anopheles hancocki (0.63%). Approximately 18 percent of these Anopheles species were infected with Plasmodium spp. Anopheles funestus, known to be involved in malaria transmission to humans, and An. moucheti-like, recently discovered in Gabon, and whose status in Plasmodium transmission is not yet elucidated, were found to be infected with great ape Plasmodium.DiscussionOur results raise the question of the potential switch of simian malaria parasites to humans. If these observations are confirmed in the future, and the infective capacity of the bridge vectors is demonstrated, this new situation could ultimately constitute an obstacle to progress in the fight against malaria.

Characterization of the complete mitochondrial genome of Culex vishnui (Diptera: Culicidae), one of the major vectors of Japanese encephalitis virus.

Culex mosquitoes (Diptera: Culicidae) can transmit a variety of arthropod-borne viruses (arboviruses), causing human and animal diseases. Cx. vishnui, Cx. pseudovishnui, and Cx. tritaeniorhynchus are three representative species in Culex vishnui subgroup, which are widely distributed in southeast Asia, and they have been proved as the main vectors transmitting Japanese encephalitis virus (JEV) that could cause human infectious mosquito-borne disease across Asia. However, the epidemiology, biology, and even molecular information of those mosquitos remain poorly understood, and only the mitochondrial genome (mitogenome) of Cx. tritaeniorhynchus has been reported in these species. In the present study, we sequenced and annotated the complete mitogenome sequence of Cx. vishnui which was 15,587bp in length, comprising 37 genes. Comparisons of nucleotide and amino acid sequences between Cx. vishnui and Cx. tritaeniorhynchus revealed that most genes within Culex vishnui subgroup were conserved, except atp8, nad1, atp6, and nad6, with differences of 0.4 (rrnS) - 15.1% (tRNAs) and 0 (nad4L) - 9.4% (atp8), respectively, interestingly suggesting the genes nad4L and rrnS were the most conserved but atp8 gene was the least. The results based on nucleotide diversity also supported a relatively uniform distribution of the intraspecific differences in Cx. vishnui and Cx. tritaeniorhynchus with only one highly pronounced peak of divergence centered at the control region. Phylogenetic analyses using concatenated amino acid sequences of 13 protein-coding genes supported the previous taxonomic classification of the family Culicidae and the monophyly of tribes Aedini, Culicini, Mansoniini, and Sabethini. The present study revealed detailed information on the subgroup Culex vishnui, reanalyzed the relationships within the family Culicidae, provided better markers to identify and distinguish Culex species, and offered more markers for studying the molecular epidemiology, population genetics, and molecular phylogenetics of Cx. vishnui.

Species composition and ecology of blood-sucking mosquitoes (Diptera, Culicidae) in Yakutia

The purpose of the research is to study the species composition and ecology of mosquitoes in Yakutia.Materials and methods. The research was performed for herd horses and cattle on pastures in Central, Western and Southern Yakutia in 2005–2021. Faunistic collections were made and the number of blood-sucking mosquito imago was recorded for animals using an entomological net with removable bags, while the average abundance rate of mosquitoes was calculated per 10 net movements in a figure-eight pattern around from 10 repetitions when studying the seasonal abundance dynamics and from 5 repetitions when studying the circadian activity rhythm.Results and discussion. In Yakutia, 15 species of blood-sucking mosquitoes of the family Culicidae were found that belong to three genera: Anopheles, Culiseta, and Aedes. Biotopes occupied by mosquito larvae of the genus Aedes include 74–126 specimens/m2. The first mosquitoes were recorded near an animal bait in the first decade of May. The mass mosquito flight period was observed from the third decade of June to the middle of the second decade of July. The total mosquitofl ight period for the season was 121–124 days. The flight of blood-sucking mosquitoes on the forest pasture continues around the clock, from the third decade of June to the middle of the second decade of July.

Biodiversity and Spatial Distribution of Mosquitoes (Diptera: Culicidae) in Kurdistan Province, Western Iran.

Mosquitoes (Diptera: Culicidae) have always been considered as the vector/s of viral and parasitic diseases. This study aimed to conduct a comprehensive survey on the species composition, spatial distribution, and biodiversity indices of mosquitoes in Kurdistan Province, western Iran. This study was carried out in 10 counties of Kurdistan Province. The immature stages of mosquitoes were collected monthly from June to September. ArcGIS software was used to spatial analysis and create maps. Alpha diversity indices were calculated using the related formula. Totally, 5831 larvae belonging to the family Culicidae were collected. Twelve species were identified including: Anopheles claviger, An. maculipennis s.l, An. superpictus s.l, Culiseta. longiareolata, Cs. subochrea, Culex hortensis, Cx. mimeticus, Cx. perexiguus, Cx. pipiens, Cx. theileri, Cx. modestus and Cx. territans. Based on this analysis, the high-risk areas of the province are determined as Anopheles in the west, Culex in the north, and the Culiseta in the south of the province. Analyzing the Alpha biodiversity indices showed Baneh and Sarabad had the maximum and Bijar had the minimum mosquito biodiversity. The western counties of the province are regarded as the hotspots for anopheline mosquitos. Moreover, reporting of malaria cases in the past, bordering with Iraq and the high traffic of travelers have made these areas as potential foci for malaria transmission. So that, routine entomological inspections are proposed to detect any suspicious vector or case entrance.

Bacillus thuringiensis Cyt Proteins as Enablers of Activity of Cry and Tpp Toxins against Aedes albopictus.

Aedes albopictus is a species of mosquito, originally from Southeast Asia, that belongs to the Culicidae family and the Dipteran insect order. The distribution of this vector has rapidly changed over the past decade, making most of the temperate territories in the world vulnerable to important human vector-borne diseases such as dengue, yellow fever, zika or chikungunya. Bacillus thuringiensis var. israeliensis (Bti)-based insecticides represent a realistic alternative to the most common synthetic insecticides for the control of mosquito larvae. However, several studies have revealed emerging resistances to the major Bti Crystal proteins such as Cry4Aa, Cry4Ba and Cry11Aa, making the finding of new toxins necessary to diminish the exposure to the same toxicity factors overtime. Here, we characterized the individual activity of Cyt1Aa, Cry4Aa, Cry4Ba and Cry11Aa against A. albopictus and found a new protein, Cyt1A-like, that increases the activity of Cry11Aa more than 20-fold. Additionally, we demonstrated that Cyt1A-like facilitates the activity three new Bti toxins: Cry53-like, Cry56A-like and Tpp36-like. All in all, these results provide alternatives to the currently available Bti products for the control of mosquito populations and position Cyt proteins as enablers of activity for otherwise non-active crystal proteins.

Relative Abundance, Distribution, and Diversity of Mosquitoes in District Peshawar, Charsadda, and Swabi

Aims: To date, there are 3601 named mosquito species recognized worldwide representing the diverse and medically important fly family Culicidae. In this study, after collection and identification of mosquitoes from the study area, their diversity, relative abundance (RA), and distribution (C) were evaluated for the prevention of mosquito-borne diseases.
 Place and Duration of Study: Mosquitoes in their immature and adult stages were collected from their habitats in district Peshawar, Charsadda, and Swabi from September 2021 till April 2022.
 Study Design: The immature samples were collected from various places such as pots, small water containers, bushy places under trees, and from the edges of some open drains. While the adult mosquitoes were collected during the same period by using the light trap and Pyrethrum spray methods for outdoor and indoor environment, respectively.
 Methodology: 252 and 295 mosquitoes in their larval and pupal, and adult stages, respectively, were collected and brought to the laboratory for rearing and were observed on daily basis. All the adult specimens were preserved using silica gel in test tubes. The specimens were kept in entomological boxes after rearing and preservation for identification. Taxonomic identification of the adult mosquitoes was made using the key provided in the fauna of British India.
 Results: The identification showed the presence of four species belonging to three genera: Culex quinquefasciatus; Aedes aegypti, Aedes albopictus; and Armigeres subalbatus. Culex quinquefasciatus was the most abundant specie found at all the study sites (C=100% and RA=44.24%), while Armigeres subalbatus was found to be an infrequent specie (C=25% and RA=20.66%). Several indices were also calculated to find out the diversity of the mosquitoes. The Shannon-Weiner index (H) for the month of April and December are 1.32 and 0.60, respectively, representing highest and lowest diversities among the study period.
 Conclusion: The study found that Culex quinquefasciatus, Aedes aegypti, and Aedes albopictus are the most prevalent species in the study area and their presence increases the risk of mosquito-borne diseases such as West Nile virus, Filariasis, and Japanese Encephalitis transmission in the area. The study suggests further research to examine the diversity of mosquitoes more thoroughly to design effective vector control strategies and predict the risk of future disease outbreaks.

Serological Determination of West Nile Virus in Domestic Birds from Rapa Nui, Chile.

Background: Flaviviruses are agents with high zoonotic potential of importance to human health. They are transmitted by mosquitoes of the Culicidae family, and birds act as host-amplifiers. Birds, mammals, and humans are susceptible hosts to infection. Methods: In this study, West Nile virus (WNV), flavivirus, infection was studied in 37 serum samples from 22 hens on Easter Island, Chile. Results: WNV was detected by ELISA (ID Screen® West Nile Competition Multi-Species). We report absence of antibodies to WNV, and to related viruses of the Japanese Encephalitis Virus serocomplex, and, therefore, absence of infection across the sample. Conclusion: This is the first evaluation of its type carried out in Chile, and represents a positive result for public health at Easter Island.

Detection of genetic material of causative agents of animal viral diseases in blood-sucking dipterans from the Tyumen Region

The purpose of the research is to test blood-sucking dipterans collected in the Tyumen Region for genetic material of viruses that cause dangerous diseases in live-stock animals.Materials and methods. From May to October 2021, blood-sucking insects of the Diptera order whose species membership was established by tabular keys were collected on pastures and farms in ten Tyumen Region districts. In 60 samples formed from the captured insects according to the taxonomic affiliation and the period and place of collection, the presence of genetic material of the leukemia provirus and the dermatitis nodularis virus of cattle (bovine) and the African swine fever virus was evaluated by polymerase chain reaction (PCR) in real time.Results and discussion. Adult insects collected for the PCR analysis were blood-sucking flies (family Muscidae, genus Stomoxys), mosquitoes (family Culicidae, genus Aedes), midges (family Simuliidae, genera Byssodon and Schoenbaueria), horseflies (family Tabanidae, genera Hybomitra, Tabanus and Haematopota), and biting midges (family Ceratopogonidae, genus Culicoides). As a result of the PCR testing of the samples for the bovine leukemia provirus DNA, 1 out of 13 samples of Stomoxys spp. (7.7%) and 1 of 13 samples of Hybomitra spp. (7.7%) were positive. The bovine leukemia provirus DNA detected in blood-sucking insects indicates the presence of this pathogen in the insect collection area as well as their possible involvement in its spread. Further research is needed on the Stomoxys spp. and Hybomitra spp. vector competence in vivo, considering natural and climatic features of the Tyumen Region.

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.

Revisiting the Evolutionary History of Culex molestus and Culex pipiens: Insights Beyond WWII Origin Narratives

Culex is a genus of mosquitoes belonging to the family Culicidae. These species are the primary vectors for diseases like the West Nile virus, St. Louis encephalitis, lymphatic filariasis, and agricultural pests. The species are distributed all over the world with overlapping species distribution. To date, it is assumed that the Culex molestus ecotype originated and evolved in London underground tunnels during WWII, when people took shelter. We tested this hypothesis by constructing phylogenetic trees containing different ecotypes and species of Culex. We charted the ecological range of C.pipiens and C.molestus and investigated the evolutionary history of the Culex species using the CO1, IGS & ITS2 gene sequences. We find some indication that C.pipiens and C.molestus form a species complex at best. In all the phylogenetic trees we constructed, C.pipiens and C.molestus shared the same ancestor and showed long branch lengths indicative of a long evolutionary history. This indicates that C.molestus could not have evolved in London tunnels during WWII. C.pipiens in all the trees showed longer branch lengths as compared to C.molestus. Our study supports the concept that evolution selects from an already existing pool of genetic variants to evolve different ecotypes and species.

Sequencing and description of the complete mitochondrial genome of Limatus durhamii (Diptera: Culicidae)

The genus Limatus (Diptera: Culicidae) are wild mosquitoes belonging to the Sabethini tribe that occurs in tropical countries and is related to transmission cycles of Orthobunyavirus (Bunyaviridae), particularly in the Amazon region. Given the unavailability of information related to evolutionary biology and molecular taxonomy aspects of this genus, we report here the first complete sequencing of the mitochondrial genome of Limatus durhamii Theobald, 1901. The NextSeq 500 platform was used for sample sequencing, and the mitochondrial sequence obtained was 14,875 bp long, comprising 37 functional subunits (13 PCGs, 22 tRNA and 02 rRNA). The phylogeny reconstructed by maximum likelihood based on the concatenation of all 13 PCGs corroborated the known taxonomic classification based most on aspects of the external morphology and few molecular studies. The data and information produced here may be useful in the future development of taxonomic and evolutionary studies for the genus, as well as the Culicidae family itself.