Interactive Effects of Salinity and Mosquito Larvicides Toxicity to Larvae of Aedes Taeniorhynchus.

Understanding the influence of salinity on the efficacy of mosquito larvicides in brackish water habitats is crucial for effective salt-marsh Aedes taeniorhynchus control. This study investigated the interactive effects of salinity on the toxicity of 3 commonly used mosquito larvicides: Bacillus thuringiensis israelensis (VectoBac® 12AS), spinosad (Natular® SC), and S-methoprene (Altosid® 12AS) against Ae. taeniorhynchus larvae. Four salinity levels (0 ppt [parts per thousand], 8 ppt, 16 ppt, and 32 ppt) were tested in laboratory bioassays. The results revealed distinct responses of these larvicides to varying salinity levels. VectoBac 12AS displayed consistent efficacy across all salinity levels, indicating its suitability for brackish water habitats. In contrast, Natular 2EC exhibited increased effectiveness with higher salinity, making it a preferable choice for saline environments. Altosid 12AS showed its highest efficacy in freshwater, with reduced effectiveness as salinity increased. These findings underscore the need to consider salinity levels when selecting and applying mosquito larvicides in diverse aquatic habitats. Understanding the complex interplay between salinity and larvicide performance is essential for optimizing mosquito control strategies and mitigating mosquito-borne diseases in various environments.

Detection of Aedes Scapularis In Southwestern Florida.

Aedes scapularis has recently been detected for the first time in southwestern Florida. During the course of research and surveillance activities by local mosquito control districts in 2020 and 2021, a total of 190 adult females were collected from 14 separate locations in Collier and Lee Counties. To date, Ae. scapularis has been found in 5 counties since its rediscovery in Florida in 2006. Its detection and likely northward expansion into Collier and Lee Counties from the southern Florida Peninsula is in line with ecological niche model predictions that found the environment of the Gulf Coast of southwestern Florida to be highly suitable for the species. Due to its potential ability to transmit both exotic and endemic pathogens such as Everglades virus and Dirofilaria immitis, understanding the range and distribution of Ae. scapularis should be a priority for Florida mosquito control and public health agencies.

Wing Interferential Patterns (WIPs) and machine learning for the classification of some Aedes species of medical interest.

Hematophagous insects belonging to the Aedes genus are proven vectors of viral and filarial pathogens of medical interest. Aedes albopictus is an increasingly important vector because of its rapid worldwide expansion. In the context of global climate change and the emergence of zoonotic infectious diseases, identification tools with field application are required to strengthen efforts in the entomological survey of arthropods with medical interest. Large scales and proactive entomological surveys of Aedes mosquitoes need skilled technicians and/or costly technical equipment, further puzzled by the vast amount of named species. In this study, we developed an automatic classification system of Aedes species by taking advantage of the species-specific marker displayed by Wing Interferential Patterns. A database holding 494 photomicrographs of 24 Aedes spp. from which those documented with more than ten pictures have undergone a deep learning methodology to train a convolutional neural network and test its accuracy to classify samples at the genus, subgenus, and species taxonomic levels. We recorded an accuracy of 95% at the genus level and > 85% for two (Ochlerotatus and Stegomyia) out of three subgenera tested. Lastly, eight were accurately classified among the 10 Aedes sp. that have undergone a training process with an overall accuracy of > 70%. Altogether, these results demonstrate the potential of this methodology for Aedes species identification and will represent a tool for the future implementation of large-scale entomological surveys.

Seasonal Abundance of Aedes Sollicitans and Aedes Taeniorhynchus Related to Temperature, Rainfall and Tidal Levels in Northeastern Florida.

The Anastasia Mosquito Control District, which manages mosquitoes in St. Johns County in northeastern Florida, has observed that the maximum numbers of the salt marsh mosquitoes, Aedes taeniorhynchus and Ae. sollicitan appeared to shift or change relative to each other, as evidenced by the Centers for Disease Control and Prevention (CDC) light trap data in the past 17 years. The aim of this study was to analyze environmental data to identify and explore these changes. Data from CDC light traps, temperature, rainfall, and tidal levels were analyzed using ANOVA. Analyses showed the 2 species had maximum abundance at different temperatures, which translated into seasonal differences with peaks of Ae. taeniorhynchus in the summer and, to a lesser extent, later in the year, and Ae. sollicitans with a peak in the autumn. This seasonal pattern was reflected in rainfall (more rain in autumn than in summer) and also, in the general area, in tidal levels (mean highest tide levels at the recording station were in autumn). The research demonstrated that simplifying the mosquito data, initially using only very high trap numbers (Mean ± 2 SD) that are important for control, identified, and made the seasonal pattern very obvious. The pattern was also observed using all the data but, although significant, was not as clear. Having identified tide as a potential driving variable, further research needs to detail spatial tidal patterns to identify areas and timing of flooding and explore the relationship between salinity and mosquito species and abundance. This is important as sea levels rise and climate changes, both potentially changing the mosquito situation and affecting control actions.

Seasonality of Mosquitoes in North Dakota, 2003-2006, with Four New State Records.

Thirty-seven species and subspecies of mosquitoes were identified from 3,580,610 specimens collected in eastern (Cass, Nelson, and Richland counties) and western (Williams County) North Dakota in 2003-2006. Four species were new state records (Aedes schizopinax, Psorophora ciliata, Ps. ferox, and Ps. horrida). Aedes vexans was dominant (82.9%). Other relatively abundant species were Ae. trivittatus (7.7%), Ae. melanimon (2.7%), Culex tarsalis (2.6%), Ae. dorsalis (1.6%), Ae. sticticus (1.0), and Culiseta inornata (0.9%). The seasonality of the species is presented.

Entomological surveillance of invasive Aedes mosquitoes in Mazandaran Province, northern Iran from 2014 to 2020.

Mosquitoes are the most important vectors of serious infectious diseases in the world. Dengue, Zika, chikungunya and yellow fever are emerging and re-emerging infectious diseases, associated with the distribution of two key vectors i.e. Aedes aegypti and Aedes albopictus throughout the world including countries neighbouring Iran. Entomological surveillance was planned and performed monthly from May to December during 2014-2020 in selected counties of Mazandaran Province, North of Iran, by ovitrap, larval collection, hand catch and human baited trap. Overall, 4410 Aedes specimens including 2376 larvae (53.9%) and 2034 (46.1%) adults belonging to six species, namely Aedes vexans, Aedes geniculatus, Aedes caspius, Aedes echinus, Aedes pulcritarsis and Aedes flavescence were collected and morphologically identified. Over the seven years of surveillance, Ae. aegypti and Ae. albopictus were not found by any sampling method. Aedes vexans and Ae. geniculatus were the most abundant species, their populations peaked in October and November and was positively correlated with precipitation and relative humidity. Aedes flavescence was a new species record for the province. A flowchart for planning and implementation of invasive mosquito surveillance for provincial health authorities in the country is proposed. These surveillance efforts provide basic and timely information for the health system to act promptly on integrated and intensified surveillance and control programs should Ae. aegypti and Ae. albopictus detected in the province.

Jamestown Canyon virus (Bunyavirales: Peribunyaviridae) vector ecology in a focus of human transmission in New Hampshire, USA.

Jamestown Canyon virus disease (JCVD) is a potentially neuroinvasive condition caused by the arbovirus Jamestown Canyon virus (JCV). Human cases of JCVD have increased in New Hampshire (NH) over the past decade, but vector surveillance is limited by funding and person power. We conducted mosquito surveillance with a focus on human JCVD cases south central NH during 2021. Routine surveillance with CDC miniature traps baited with CO2 (lights removed) was supplemented by a paired trapping design to test the collection efficiency of octenol, and New Jersey light traps. We performed virus testing, blood meal analysis, and compared morphological identification with DNA barcoding. Over 50,000 mosquitoes were collected representing 28 species. Twelve JCV-positive pools were derived from 6 species of more than 1,600 pools tested. Of those, Aedes excrucians/stimulans (MLE 4.95, Diptera: Culicidae, Walker, 1856, 1848), and Aedes sticticus (MLE 2.02, Meigen, 1838) had the highest JCV infection rates, and Aedes canadensis (MLE 0.13, Theobold, 1901) and Coquillettidia perturbans (0.10, Diptera: Culicidae, Walker, 1856) had the lowest infection rates. One hundred and fifty-one blood meals were matched to a vertebrate host. All putative vectors fed on the amplifying host of JCV, white-tailed deer (36-100% of bloodmeals). Putative vectors that fed on human hosts included Aedes excrucians (8%), Anopheles punctipennis (25%, Diptera: Culicidae, Say, 1823), and Coquillettidia perturbans (51%). CDC traps baited with CO2 were effective for collecting putative vectors. DNA barcoding enhanced morphological identifications of damaged specimens. We present the first ecological overview of JCV vectors in NH.

Haemagogus equinus in Kinney County, Texas.

Larvae collected from an oak tree hole east of Brackettville, Kinney County, Texas, resulted in the collection of 3 species: Aedes triseriatus, Ae. zoosophus, and Haemagogus equinus. One larva and 1 female of Hg. equinus were recovered from this sample. This is the first report of Hg. equinus in Kinney County and the first report of this species being collected in Texas since 1962. Samples collected from other sites documented the presence of Ae. epactius, Culex nigripalpus, and Psorophora signipennis in Kinney County.

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.

New Records of Mosquitoes in Bolivia and Northwestern Argentina.

Two mosquito species, Culex (Culex) hepperi and Culex (Culex) maxi are reported for the first time for Bolivia, in Tarija department. Aedes (Ochlerotatus) stigmaticus and Mansonia (Mansonia) indubitans are the first records in Aniceto Arce province, in Tarija department, southern Bolivia. In addition, Aedes (Protomacleaya) alboapicus and Culex (Phytotelmatomyia) renatoi are reported for the first time in the northwestern region of Argentina. Anopheles (Nyssorrhynchus) nuneztovari s.l. is reported for the first time in Jujuy and Tucumán provinces, and Aedes (Ochlerotatus) raymundi and Anopheles (Anopheles) neomaculipalpus are the first reports in Tucumán province. The 3 species are extending their geographical distribution in these provinces. Data on the collection localities and comments about the medical relevance of some species are also presented.

Aedes pertinax, Anopheles perplexens, Culex declarator, and Cx. interrogator: An Update of Mosquito Species Records for Charlotte County, Florida.

Understanding the geographic occurrence of mosquito species is an important element to addressing public health and nuisance mosquito-related issues, particularly as changing climates and increased global connectivity is likely to facilitate changes in the distribution of mosquitoes and other species. In Charlotte County, FL, routine surveillance of mosquito species for public health in 2019-21 identified 4 mosquito species not previously documented in the county. Aedes pertinax, Anopheles perplexens, Culex declarator, and Cx. interrogator adults were collected and verified to species level. Aedes pertinax and Cx. declarator and were collected in 2019, whereas An. perplexens and Cx. interrogator were documented from collections in 2021. All 4 species were initially visually identified by external morphology and confirmed by sequencing the DNA barcoding region of the cytochrome c oxidase subunit I gene. Apart from native An. perplexens, in which only 1 specimen has been confirmed to date, the 3 newly documented nonnative species are now recognized throughout the county.

Aedes pertinax, A Lost and Found New Species Record for Monroe County, Florida.

Aedes pertinax, a little-studied tropical mosquito that has found its way into the USA, has recently been discovered to have existed in the Florida Keys for almost 25 years. A 2021 collection of Ae. pertinax adult females in Key Largo, FL, sparked a retrospective search for stored specimens collected in 1998 on Big Pine Key, FL. Positive identification of the specimens from the 1998 collection confirmed the specimens to be Ae. pertinax, predating the first reported specimen in the USA by 13 years.

A deeper insight into the sialome of male and female Ochlerotatus triseriatus mosquitoes.

Over the last 20 years, advancements in sequencing technologies have highlighted the unique composition of the salivary glands of blood-feeding arthropods. Further biochemical and structural data demonstrated that salivary proteins can disrupt host hemostasis, inflammation and immunity, which favors pathogen transmission. Previously, a Sanger-based sialome of adult Ochlerotatus triseriatus female salivary glands was published based on 731 expressed sequence tag (ESTs). Here, we revisited O. triseriatus salivary gland contents using an Illumina-based sequencing approach of both male and female tissues. In the current data set, we report 10,317 DNA coding sequences classified into several functional classes. The translated transcripts also served as a reference database for proteomic analysis of O. triseriatus female saliva, in which unique peptides from 101 proteins were found. Finally, comparison of male and female libraries allowed for the identification of female-enriched transcripts that are potentially related to blood acquisition and virus transmission.

Insecticide Susceptibility Tests of Aedes taeniorhynchus in Yucatan, Mexico.

We report results of susceptibility tests for Aedes taeniorhynchus from 2 localities of Yucatan State, Mexico, to different insecticides. The Centers for Disease Control and Prevention bottle bioassays were performed using the active ingredients of 3 pyrethroids, 2 organophosphates, and 1 carbamate: permethrin (15 µg/ml), deltamethrin (10 µg/ml), alpha-cypermethrin (10 µg/ml), malathion (50 µg/ml), chlorpyrifos (85 µg/ml), and bendiocarb (12.5 µg/ml). The mortality recorded at the diagnostic time of exposure (30 min) was 100% with all insecticides evaluated and for both populations. These results suggest complete susceptibility to the 3 chemical groups generally used for urban Ae. aegypti mosquito control.

Characterisation of the RNA Virome of Nine Ochlerotatus Species in Finland.

RNA viromes of nine commonly encountered Ochlerotatus mosquito species collected around Finland in 2015 and 2017 were studied using next-generation sequencing. Mosquito homogenates were sequenced from 91 pools comprising 16-60 morphologically identified adult females of Oc. cantans, Oc. caspius, Oc. communis, Oc. diantaeus, Oc. excrucians, Oc. hexodontus, Oc. intrudens, Oc. pullatus and Oc. punctor/punctodes. In total 514 viral Reverse dependent RNA polymerase (RdRp) sequences of 159 virus species were recovered, belonging to 25 families or equivalent rank, as follows: Aliusviridae, Aspiviridae, Botybirnavirus, Chrysoviridae, Chuviridae, Endornaviridae, Flaviviridae, Iflaviridae, Negevirus, Partitiviridae, Permutotetraviridae, Phasmaviridae, Phenuiviridae, Picornaviridae, Qinviridae, Quenyavirus, Rhabdoviridae, Sedoreoviridae, Solemoviridae, Spinareoviridae, Togaviridae, Totiviridae, Virgaviridae, Xinmoviridae and Yueviridae. Of these, 147 are tentatively novel viruses. One sequence of Sindbis virus, which causes Pogosta disease in humans, was detected from Oc. communis from Pohjois-Karjala. This study greatly increases the number of mosquito-associated viruses known from Finland and presents the northern-most mosquito-associated viruses in Europe to date.

First national record of Aedes melanimon (Diptera: Culicidae) in Mexico.

Aedes (Ochlerotatus) melanimon Dyar 1924 has been considered an important pest in agricultural and rural communities. Aedes melanimon is a vector of WEEV and CEV and is a competent laboratory vector of WNV. The known range of Ae. melanimon extends throughout Southwest Canada, part of Central and most of Western USA. Here we report the first record of Ae. melanimon in Mexico, at Ciudad Juárez, Chihuahua. The collect represents the southernmost distributional record for this species. Its indigenous presence in a highly urbanized and dry area was not expected. A permanent surveillance program to detect and determine the species in border-crossing cities is encouraged.

First description of the mitogenome and phylogeny:Aedes vexansand Ochlerotatus caspius of the Tribe Aedini (Diptera: Culicidae).

Culicidae, the mosquito family, includes more than 3600 species subdivided into the subfamilies Anophelinae and Culicinae. One-third of mosquitoes belong to the Aedini tribe, which is subordinate to the subfamily Culicinae, which comprises common vectors of viral zoonoses. The tribe of Aedini is extremely diverse in morphology and geographical distribution and has high ecological and medical significance. However, knowledge about the systematics of the Aedini tribe is still limited owing to its large population and the similar morphological characteristics of its species. This study provides the first description of the complete mitochondrial (mt) genome sequence of Aedes vexans and Ochlerotatus caspius belonging to the Aedini tribe. The mt genomes of A. vexans and O. caspius are circular molecules that are 15,861 bp and 15,954 bp in size, with AT contents of 78.54% and 79.36%, respectively. Both the circular mt genomes comprise 37 functional subunits, including 13 protein-coding genes (PCGs), two ribosomal RNA genes, 22 transfer RNA genes (tRNAs), and a control region (also known as the AT-rich region). The most common start codons are ATT/ATG, apart from cox1 (TCG) and nad5 (GTG), while TAA is the termination codon for all PCGs. All tRNAs have a typical clover leaf structure, except tRNA Ser1. Phylogenetic analysis of the concatenated, aligned amino acid sequences of the 13 PCGs showed that A. vexans gathered with Aedes sp. in a sister taxon, and O. caspius gathered with Ochlerotatus sp. in a sister taxon. The findings from the present study support the concept of monophyly of all groups, ratify the current taxonomic classification, and provide vital molecular marker resources for further studies of the taxonomy, population genetics, and systematics of the Aedini tribe.

Rediscovering the mosquito fauna (Diptera: Culicidae) of Estonia: an annotated checklist with distribution maps and DNA evidence.

Female mosquitoes (Diptera: Culicidae) affect their hosts in numerous negative ways and are crucial to the spread of vector-borne pathogens. It is, therefore, important to have a detailed overview of regional mosquitoes, to be able to detect changes in species diversity and identify possible health threats. The aim of this study was to update the checklist of the mosquito fauna of Estonia for the first time since 1957. For this purpose, 24,344 adult mosquitoes (94% females) were collected in Estonia from 2008 to 2020 using various trapping methods. Specimens were primarily identified by morphological characteristics, but DNA barcoding based on the partial cytochrome c oxidase subunit 1 gene (COI) was also used. Species were included in the checklist based on historical records as well as new collections, while also considering reports from neighboring countries. Species records are supplemented with voucher specimens, distribution maps and DNA evidence. The updated checklist includes 34 species, 27 of which were confirmed with recently collected material. All in all, Aedes communis (de Geer, 1776) proved to be the most common mosquito in Estonia, accounting for 30.1% of the specimens collected. This is noteworthy, as this species has been implicated in the transmission of multiple disease agents present in the area. New evidence revealed the presence of Ae. hexodontus Dyar, 1916, Ae. sticticus (Meigen, 1838), Anopheles messeae Falleroni, 1926 and Culiseta bergrothi (Edwards, 1921) in Estonia.

Using a pupal exuvia to designate the undamaged neotype of a species belonging to a complex of sibling species - the case of Aedes coluzzii (Diptera, Culicidae).

The mosquito species Aedes (Ochlerotatus) coluzzii Rioux, Guilvard & Pasteur, 1998 was distinguished from its sibling species Aedes detritus (Haliday, 1833) using an isoenzymatic method that required the destruction of the entire specimen, therefore no holotype was designated by the species authors. We aimed to designate a neotype for Ae. coluzzii from specimens collected from the type-locality and individually reared up to adult stage. Genomic DNA was extracted from pupal exuvia and ITS2 was sequenced, enabling verification of the identity of each specimen as Ae. coluzzii or Ae. detritus. Among the series of Ae. coluzzii, a male was designated as neotype and deposited in a collection. To our knowledge, this is the first time the type of a mosquito species is deposited thanks to its molecular identification from its pupal exuvia. The set of identified specimens allowed additional phylogenetic and morphologic studies.

Title: Utilisation d'une exuvie nymphale pour désigner le néotype intact d'une espèce appartenant à un complexe d'espèces jumelles - le cas d'Aedes coluzzii (Diptera, Culicidae). Abstract: L'espèce de moustique Aedes (Ochlerotatus) coluzzii Rioux, Guilvard & Pasteur, 1998 a été distinguée de son espèce jumelle Aedes detritus (Haliday, 1833) par une méthode isoenzymatique qui a nécessité la destruction de l'ensemble du spécimen, et donc aucun holotype n'a été désigné par les auteurs de l'espèce. Notre objectif était de désigner un néotype pour Ae. coluzzii à partir de spécimens collectés dans la localité-type et élevés individuellement jusqu'au stade adulte. L'ADN génomique a été extrait de l'exuvie nymphale et l'ITS2 a été séquencé, permettant la vérification de l'identité de chaque spécimen comme Ae. coluzzii ou Ae. détritus. Parmi la série d'Ae. coluzzii, un mâle a été désigné comme néotype et déposé dans une collection. À notre connaissance, c'est la première fois que le type d'une espèce de moustique est déposé grâce à l'identification moléculaire à partir de son exuvie nymphale. L'ensemble des spécimens identifiés a permis des études phylogénétiques et morphologiques complémentaires.

First records of Aedes pulcritarsis (Rondani, 1872) (Diptera: Culicidae) in Austria.

Aedes pulcritarsis is a tree-hole breeding species with its main distribution in the Mediterranean area. Within the scope of two independent monitoring programmes, this mosquito species was detected for the first time in Austria, in the province of Lower Austria (2018, districts Mistelbach and Gaenserndorf; 2020, district Bruck an der Leitha). As the climatic and habitat situation in Central Europe seems to be generally suitable for this species, the most likely explanation for the species not being recorded previously is that it might have been overlooked in the past due to its specialized breeding habitat. However, further research on the distribution of Ae. pulcritarsis in Austria would be needed to support this hypothesis. The results from this study will contribute to the investigation of the northern distribution limit of Ae. pulcritarsis in Europe and possible changes thereof.