Geometric morphometrics to differentiate species and explore seasonal variation in three Mansonia species (Diptera: Culicidae) in central Thailand and their association with meteorological factors.

Mansonia mosquito species are recognised as a significant vector of human pathogens, primarily transmitting the filarial nematode, Brugia malayi. In central Thailand, the three most prevalent Mansonia species are Mansonia annulifera, Mansonia indiana and Mansonia uniformis. This study explored the influence of seasonal changes on the phenotypic variation of these Mansonia species in central Thailand using the geometric morphometrics (GM). To ensure accurate species identification, we integrated GM techniques with DNA barcoding, examining distinctions in both phenotype and genotype among the species. The intraspecific genetic divergence ranged from 0.00% to 1.69%, whereas the interspecific genetic divergence ranged from 10.52% to 16.36%. The clear distinction between intra- and interspecific distances demonstrated the presence of a barcoding gap, confirming the successful differentiation of the three Mansonia mosquito species through DNA barcoding. Similarly, the interspecies GM assessment for classifying Mansonia species demonstrated a high degree of accuracy, with an overall performance of 98.12%. Exploring seasonal variation in the three Mansonia species revealed wing variations across different seasons, and pronounced variations appearing in the cool season. Regarding their association with meteorological factors, Ma. annulifera and Ma. uniformis showed significant positive correlations with temperature (p < 0.05), and Ma. uniformis also displayed a significant negative correlation with atmospheric pressure (p < 0.05). The insights from this study will deepen our understanding of the adaptive patterns of Mansonia mosquitoes in Thailand's central region, paving the way for enhanced disease surveillance related to these vectors.

Influence of insular conditions on wing phenotypic variation in two dominant mosquito vectors, Aedes albopictus and Armigeres subalbatus (Diptera: Culicidae), in the border archipelagos of Thailand.

Insects geographically separated into island and mainland populations often exhibit phenotypic variations, a phenomenon known as insular conditions. These conditions can lead to rapid evolutionary changes that affect the morphological characteristics of mosquito vectors. Nevertheless, studies that specifically examine phenotype differences between island and mainland mosquito populations have been limited. In this study, wing variation in size and shape was investigated using the geometric morphometric (GM) technique in two dominant mosquito vectors, Aedes albopictus and Armigeres subalbatus, in the Ranong and Trat archipelagos of Thailand. Significant differences in average wing centroid size (CS) were found in 6 out of 15 population pairs for Ae. albopictus (p < 0.05) and in 5 population pairs for Ar. subalbatus (p < 0.05). After removing the allometric effect, canonical variate analyses (CVA) based on wing shape analysis revealed overlap across all populations for both Ae. albopictus and Ar. subalbatus. However, the statistical analysis indicated that Ar. subalbatus exhibited wing shape differences across all populations (p < 0.05), and most Ae. albopictus populations also displayed distinct wing shapes (p < 0.05), except for the populations from Chang Island and the mainland of Ranong, which showed no significant differences (p > 0.05). These findings enhance our understanding of mosquito adaptability in island regions and provide valuable data for the surveillance and monitoring of vector evolution.

Integrating wing morphometrics and mitochondrial DNA analysis to assess the filaria vector Mansonia uniformis (Diptera: Culicidae) populations in Thailand.

Mansonia uniformis (Diptera: Culicidae) is recognized as a vector of Brugia malayi and has been reported to transmit Wuchereria bancrofti, both causing lymphatic filariasis in humans. This study employed geometric morphometrics (GM) to investigate wing shape variation and analyzed genetic diversity through cytochrome c oxidase subunit 1 (COI) gene analyses in Ma. uniformis populations across Thailand. Wing GM analyses indicated significant differences in wing shape based on Mahalanobis distances among nearly all population pairs (p < 0.05), with no significant correlation between wing shape and geographic distance (r = 0.210, p > 0.05). Genetic analyses identified 63 haplotypes and 49 polymorphic sites, with the overall population exhibiting a nucleotide diversity of 0.006 (± 0.001) and a haplotype diversity of 0.912 (± 0.017). Deviations from neutrality, as indicated by Tajima's D and Fu's FS tests for the overall Ma. uniformis populations in Thailand, were statistically significant and negative, suggesting population expansion (both p < 0.05). Analysis of molecular variance revealed no significant genetic structure when all populations were categorized based on collection sites and geographic regions. However, significant differences in FST values were observed between some populations. These findings enhance our understanding of the geographical and genetic factors influencing Ma. uniformis populations, which are crucial for developing effective control strategies in Thailand.

Dynamic flight stability of hovering mosquitoes.

The flight of mosquitoes is unusual compared with many other insects, such as fruit-flies and honey bees: mosquitoes fly with their legs spread; they also have rather short stroke amplitude, hence use different aerodynamic mechanisms to produce lift. Could their flight-stability properties be different from those of other insects? Here, we first measured wing kinematics and morphological parameters of two hovering mosquitoes, and then use the method of computational fluid dynamics to compute the aerodynamic derivatives and the techniques of eigenvalue and eigenvector analysis to study their stability properties. We found that their natural-mode structure is the same as that of many other insects: for the longitudinal motion, one unstable oscillatory mode, one stable fast subsidence mode and one stable slow subsidence mode; for the lateral motion: an unstable divergence mode, a stable oscillatory mode and a stable subsidence mode. The different aerodynamic mechanisms of mosquitoes do not change the major aerodynamic derivatives. The spread legs of mosquitoes have great effect on the moments of inertia and make the eigenvalue of the stable lateral mode much smaller. However, the leg-spreading has only a small quantitative effect on the unstable eigenvalues: the magnitudes of the eigenvalues in the two unstable modes, or the growth rate of the disturbances, are reduced by approximately 11%, compared to those calculated without considering the spread legs.

Using multilayer perceptron computation to discover ideal insect olfactory receptor combinations in the mosquito and fruit fly for an efficient electronic nose.

The model organism, Drosophila melanogaster, and the mosquito Anopheles gambiae use 60 and 79 odorant receptors, respectively, to sense their olfactory world. However, a commercial "electronic nose" in the form of an insect olfactory biosensor demands very low numbers of receptors at its front end of detection due to the difficulties of receptor/sensor integration and functionalization. In this letter, we demonstrate how computation via artificial neural networks (ANNs), in the form of multilayer perceptrons (MLPs), can be successfully incorporated as the signal processing back end of the biosensor to drastically reduce the number of receptors to three while still retaining 100% performance of odorant detection to that of a full complement of receptors. In addition, we provide a detailed performance comparison between D. melanogaster and A. gambiae odorant receptors and demonstrate that A. gambiae receptors provide superior olfaction detection performance over D. melanogaster for very low receptor numbers. The results from this study present the possibility of using the computation of MLPs to discover ideal biological olfactory receptors for an olfactory biosensor device to provide maximum classification performance of unknown odorants.

Old ingredients for a new recipe? Neem cake, a low-cost botanical by-product in the fight against mosquito-borne diseases.

Mosquitoes (Diptera: Culicidae) represent an important threat to millions of people worldwide, since they act as vectors for important pathogens, such as malaria, yellow fever, dengue and West Nile. Control programmes mainly rely on chemical treatments against larvae, indoor residual spraying and insecticide-treated bed nets. In recent years, huge efforts have been carried out to propose new eco-friendly alternatives, with a special focus on the evaluation of plant-borne mosquitocidal compounds. Major examples are neem-based products (Azadirachta indica A. Juss, Meliaceae) that have been proven as really effective against a huge range of pests of medical and veterinary importance, including mosquitoes. Recent research highlighted that neem cake, a cheap by-product from neem oil extraction, is an important source of mosquitocidal metabolites. In this review, we examined (i) the latest achievements about neem cake metabolomics with special reference to nor-terpenoid and related content; (ii) the neem cake ovicidal, larvicidal and pupicidal toxicity against Aedes, Anopheles and Culex mosquito vectors; (iii) its non-target effects against vertebrates; and (iv) its oviposition deterrence effects on mosquito females. Overall, neem cake can be proposed as an eco-friendly and low-cost source of chemicals to build newer and safer control tools against mosquito vectors.

Mansonia titillans: New Resident Species or Infrequent Visitor in Chatham County, Georgia, and Beaufort County, South Carolina, USA.

In September, October, and November 2014, adult Mansonia titillans were collected at 4 separate sites near Savannah in Chatham County, Georgia, and 1 site in Muscogee County, GA, during routine mosquito surveillance. Although previously recorded from Beaufort County, SC, and several inland southern Georgia counties, recent reports of this species from coastal Georgia or South Carolina are lacking. These newly captured Ma. titillans specimens represent the first documented records for Muscogee County and Chatham County, GA, and may indicate a recent northern expansion or reintroduction of this species along the Georgia and South Carolina coast.

[Development of a Futuristic Microneedle Based on Nanotechnology].

The author proposes "the microtube production technique", which is based on "nanotechnology", to produce a microneedle which mimicks a female mosquito's labium to collect blood almost painlessly. The microneedle production technique is to deposit the microneedle material on a rotating wire substrate at a speed of 3-5 rpm by a sputtering deposition method and etch the wire substrate by a chemical solution after a heat treatment As the result, a titanium microneedle was produced (outer and inner diameter: 50 µm and 25 µm) in the same size of female mosquito's labium. In this paper, microtube production technique is introduced first as a key technology. The design technique of painless microneedle for blood extraction system based on quantitative pain assessment, are also proposed to produce solid type microneedle with trenches on the surface of the needle.

Phase retrieval using spatially modulated illumination.

In this Letter, we propose a method for retrieving the phase of a wavefront from the diffraction patterns recorded when the object is sequentially illuminated by spatially modulated light. For wavefronts having a smooth phase, the retrieval is achieved by using a deterministic method. When the phase has discontinuities, an iterative process is used for the retrieval and enhancement of the spatial resolution. Both the deterministic and iterative phase reconstructions are demonstrated by experiments.

A new subgenus and species of Topomyia (Diptera: Culicidae: Sabethini) based on a remarkable male mosquito from Sabah, Malaysia.

Miyagiella Harbach, subgen. nov., is introduced as a new subgenus of Topomyia Leicester for a remarkable male mosquito, Topomyia discors Harbach, sp. nov., from Sabah, Malaysia. A diagnosis of the subgenus is provided that features unique anatomical characters of the genitalia of the holotype male. Miyagiella is very distinct from the two previously recognised subgenera of Topomyia, but is perhaps more closely related to the nominotypical subgenus than to subgenus Suaymyia Thurman. Salient differences that distinguish the three subgenera are contrasted; the holotype male of To. discors is described and its unique genitalia are illustrated.

Morphological characterization of antennal sensilla in Toxorhynchites theobaldi, Toxorhynchites violaceus, and Lutzia bigoti adults: a comparative study using scanning electron microscopy.

Some mosquitoes, including species of the genus Toxorhynchites, are known for actively preying on other mosquito larvae, making these predators valuable allies in the fight against vector-borne diseases. A comprehensive understanding of the anatomy and physiology of these potential biological control agents is helpful for the development of effective strategies for controlling vector populations. This includes the antennae, a crucial component in the search for hosts, mating, and selection of oviposition sites. This study utilized scanning electron microscopy to characterize the sensilla on the antennae of adult mosquitoes from two species that are exclusively phytophagous, including Toxorhynchites theobaldi and Toxorhynchites violaceus, as well as Lutzia bigoti, which females are allegedly hematophagous. The types of sensilla in each species were compared, and five basic types of antennal sensilla were identified: trichoid, chaetic, coeloconic, basiconic, and ampullacea. The analysis also found that they were morphologically similar across the three species, regardless of feeding habits or sex. The identification and characterization of basic types of antennal sensilla in T. theobaldi, T. violaceus, and L. bigoti suggest that these structures, which play a crucial role in the behavior and ecology, have common functions across different mosquito species, despite differences in feeding habits or sex.

Midgut morphology of the predator mosquito Lutzia bigoti (Diptera: Culicidae) and its implications for feeding behavior.

Lutzia mosquitoes (Theobald, 1903) are predaceous during their larval stages, but the adult feeding is not clearly understood, especially in relation to blood feeding. In case these mosquitoes are harmless to humans and related animals, they can be useful in biological control of mosquito vectors of pathogens. Investigating the midgut morphology is a good strategy to understand the feeding behavior of this species. The midgut in Lutzia bigoti Bellardi, 1862 displays two distinct portions, a thin anterior midgut and a more dilated posterior midgut. Digestive cells form a single epithelium in the midgut. These cells have long and packed microvilli at their apex and membrane infoldings at their basal portion, the basal labyrinth. The epithelium is supported by a basal lamina. Regarding their cytoplasm, it is noteworthy the abundance of mitochondria, distributed in an apical-basal fashion, and also a whirl-shaped endoplasmic reticulum in the posterior midgut. Basal cells are also found in the midgut of L. bigoti, resembling regenerative cells. The general organization of L. bigoti's midgut closely resembles that of numerous hematophagous mosquitoes previously documented. However, it diverges due to the presence of a peritrophic matrix even when exclusively fed on sugary solutions. Peculiar aspects of L. bigoti's midgut are discussed and compared to those of other mosquito species.

A DNA barcode reference library of Croatian mosquitoes (Diptera: Culicidae): implications for identification and delimitation of species, with notes on the distribution of potential vector species.

BACKGROUND: Mosquitoes pose a risk to human health worldwide, and correct species identification and detection of cryptic species are the most important keys for surveillance and control of mosquito vectors. In addition to traditional identification based on morphology, DNA barcoding has recently been widely used as a complementary tool for reliable identification of mosquito species. The main objective of this study was to create a reference DNA barcode library for the Croatian mosquito fauna, which should contribute to more accurate and faster identification of species, including cryptic species, and recognition of relevant vector species. METHODS: Sampling was carried out in three biogeographical regions of Croatia over six years (2017-2022). The mosquitoes were morphologically identified; molecular identification was based on the standard barcoding region of the mitochondrial COI gene and the nuclear ITS2 region, the latter to identify species within the Anopheles maculipennis complex. The BIN-RESL algorithm assigned the COI sequences to the corresponding BINs (Barcode Index Number clusters) in BOLD, i.e. to putative MOTUs (Molecular Operational Taxonomic Units). The bPTP and ASAP species delimitation methods were applied to the genus datasets in order to verify/confirm the assignment of specimens to specific MOTUs. RESULTS: A total of 405 mosquito specimens belonging to six genera and 30 morphospecies were collected and processed. Species delimitation methods assigned the samples to 31 (BIN-RESL), 30 (bPTP) and 28 (ASAP) MOTUs, with most delimited MOTUs matching the morphological identification. Some species of the genera Culex, Aedes and Anopheles were assigned to the same MOTUs, especially species that are difficult to distinguish morphologically and/or represent species complexes. In total, COI barcode sequences for 34 mosquito species and ITS2 sequences for three species of the genus Anopheles were added to the mosquito sequence database for Croatia, including one individual from the Intrudens Group, which represents a new record for the Croatian mosquito fauna. CONCLUSION: We present the results of the first comprehensive study combining morphological and molecular identification of most mosquito species present in Croatia, including several invasive and vector species. With the exception of some closely related species, this study confirmed that DNA barcoding based on COI provides a reliable basis for the identification of mosquito species in Croatia.

Resolution of cryptic mosquito species through wing morphometrics.

Mosquitoes are medically important insects, and accurate species identification is crucial to understanding vector biology, forming the cornerstone of successful vector control programs. Identification is difficult owing to morphologically similar species. Wing morphometrics can provide a simple, fast, and accurate way to classify species, and using it as a method to differentiate vector species among its cryptic congeners has been underexplored. Using a total of 227 mosquitoes and 20 landmarks per specimen, we demonstrated the utility of wing morphometrics in differentiating species two groups occurring in sympatry - Culex (Culex) vishnui group and Culex (Lophoceraomyia) subgenus, as well as explored population-level variation in the wing shape of Aedes albopictus across habitats. Cytochrome oxidase subunit I (COI) gene region was sequenced to validate the morphological and morphometric identification. Procrustes ANOVA regression and CVA based on wing shape reflected that the wing landmarks across all species differed significantly, and leave-one-out cross validation revealed an overall high accuracy of >97% for the two Culex groups. Wing morphometrics uncovered population-level variation within Aedes albopictus, but cross validation accuracy was low. Overall, we show that wing geomorphometric analysis is able to resolve cryptic Culex species (including vectors) occurring sympatrically, and is a robust tool for identifying mosquitoes reliably.

Investigating the impact of climate and seasonality on mosquito (Diptera: Culicidae) vector populations in the connecting areas of the Tenasserim range forests in Thailand.

Mosquito-borne diseases pose a significant public health challenge globally. Our study focused on the seasonal diversity of mosquito species in the connecting areas of the Tenasserim (also known as Tanaosri) range forests in Thailand. Additionally, we employed the geometric morphometric technique to assess variations in wing size and shape among five predominant mosquito species. Throughout the study period, we collected a total of 9,522 mosquitoes, encompassing 42 species across eight genera. In these connecting areas of forests, the Simpson index and Shannon species diversity index were recorded at 0.86 and 2.36, respectively, indicating a high level of mosquito diversity. Our analysis using the Analysis of Similarities (ANOSIM) test showed significant seasonal differences in mosquito communities, with an R-value of 0.30 (p < 0.05) in the lower connecting areas and 0.37 (p < 0.05) in the upper connecting areas. Additionally, canonical correspondence analyses showed that the abundance of each mosquito species is influenced by various climate factors. Phenotypic analyses of wing size and shape have deepened our understanding of local adaptation and the seasonal pressures impacting these vectors. Notably, most species exhibited larger wing sizes in the dry season compared to other seasons. Additionally, seasonal assessments of wing shape in five predominant mosquito species revealed significant differences across seasonal populations (p < 0.05). Ongoing monitoring of these populations is crucial to enhancing our understanding of the seasonal effects on mosquito abundance and physiological adaptations. These insights are essential for developing more effective strategies to manage mosquito-borne diseases.

Robust mosquito species identification from diverse body and wing images using deep learning.

Mosquito-borne diseases are a major global health threat. Traditional morphological or molecular methods for identifying mosquito species often require specialized expertise or expensive laboratory equipment. The use of convolutional neural networks (CNNs) to identify mosquito species based on images may offer a promising alternative, but their practical implementation often remains limited. This study explores the applicability of CNNs in classifying mosquito species. It compares the efficacy of body and wing depictions across three image collection methods: a smartphone, macro-lens attached to a smartphone and a professional stereomicroscope. The study included 796 specimens of four morphologically similar Aedes species, Aedes aegypti, Ae. albopictus, Ae. koreicus and Ae. japonicus japonicus. The findings of this study indicate that CNN models demonstrate superior performance in wing-based classification 87.6% (95% CI: 84.2-91.0) compared to body-based classification 78.9% (95% CI: 77.7-80.0). Nevertheless, there are notable limitations of CNNs as they perform reliably across multiple devices only when trained specifically on those devices, resulting in an average decline of mean accuracy by 14%, even with extensive image augmentation. Additionally, we also estimate the required training data volume for effective classification, noting a reduced requirement for wing-based classification compared to body-based methods. Our study underscores the viability of both body and wing classification methods for mosquito species identification while emphasizing the need to address practical constraints in developing accessible classification systems.

Phase retrieval with resolution enhancement by using structured illumination.

In this Letter, we present referenceless phase retrieval methods with resolution enhancement. Structured illuminations with different orientations and phase shifts are generated by a spatial light modulator and are used to illuminate the specimen. The generated diffraction patterns are recorded by a CCD camera, and the phase of the wavefront is reconstructed from these patterns.

A new species of the genus Orthopodomyia from Ototomo, Cameroon, with notes on other Orthopodomyia group-species.

A new species belonging to the new Ototomoensis Group, Orthopodomyia ototomoensis Huang and Rueda, is described from Ototomo, Cameroon. Diagnostic features for separating members of Nkolbissonensis Group and Ototomoensis Group from other Orthopodomyia groups are provided. Two supplemental keys are presented: "A Pictorial Key to the Species of the Genus Orthopodomyia Ototomoensis Group in the Afrotropical Region (Diptera: Culicidae)" and "A Pictorial Key to the Species of the Genus Orthopodomyia Nkolbissonensis Group in the Afrotropical Region (Diptera: Culicidae)".

Annotated checklist of the mosquitoes of the Republic of Moldova.

The mosquito fauna of the Republic of Moldova is poorly known. In an effort to understand the Culicidae fauna better, mosquito collections have been conducted between early April and middle November from 2008 to 2012. A total of 10,923 larval specimens and 8,246 adults were collected from 20 regions of Moldova. Altogether 36 species have been recorded during the recent study, bringing the total Moldovan mosquito fauna to 40 species in 9 genera and 11 subgenera. New state records include the following 7 species: Anopheles pseudopictus, An. melanoon, Aedes geminus, Culex torrentium, Culiseta longiareolata, Coquillettidia buxtoni, and Uranotaenia unguiculata.

Description of male, pupa and larva of Psorophora (Grabhamia) paulli and redescription of the female (Diptera: Culicidae).

The female of Psorophora (Grabhamia) paulli Paterson & Shannon is redescribed, and the pupa, fourth-instar larva and male genitalia are described and illustrated for the first time. Information about the distribution, bionomics and taxonomy is also included. Adults of Ps. paulli can be separated from the other species of the genus and subgenus by its small size. The larva of Ps. paulli is similar to that of Ps. varinervis Edwards and Ps. discolor (Coquillett) but can be separated based on the development of setae 1-X and 5-VIII, the length of the anal papillae and the comb on a sclerotized area.