BACKGROUND: Malaria impacts nearly 250 million individuals annually. Specifically, Uganda has one of the highest burdens, with 13 million cases and nearly 20,000 deaths. Controlling the spread of malaria relies on vector surveillance, a system where collected mosquitos are analyzed for vector species\' density in rural areas to plan interventions accordingly. However, this relies on trained entomologists known as vector control officers (VCOs) who identify species via microscopy. The global shortage of entomologists and this time-intensive process cause significant reporting delays. VectorCam is a low-cost artificial intelligence-based tool that identifies a mosquito\'s species, sex, and abdomen status with a picture and sends these results electronically from surveillance sites to decision makers, thereby deskilling the process to village health teams (VHTs).
OBJECTIVE: This study evaluates the usability of the VectorCam system among VHTs by assessing its efficiency, effectiveness, and satisfaction.
METHODS: The VectorCam system has imaging hardware and a phone app designed to identify mosquito species. Two users are needed: (1) an imager to capture images of mosquitos using the app and (2) a loader to load and unload mosquitos from the hardware. Critical success tasks for both roles were identified, which VCOs used to train and certify VHTs. In the first testing phase (phase 1), a VCO and a VHT were paired to assume the role of an imager or a loader. Afterward, they swapped. In phase 2, two VHTs were paired, mimicking real use. The time taken to image each mosquito, critical errors, and System Usability Scale (SUS) scores were recorded for each participant.
RESULTS: Overall, 14 male and 6 female VHT members aged 20 to 70 years were recruited, of which 12 (60%) participants had smartphone use experience. The average throughput values for phases 1 and 2 for the imager were 70 (SD 30.3) seconds and 56.1 (SD 22.9) seconds per mosquito, respectively, indicating a decrease in the length of time for imaging a tray of mosquitos. The loader\'s average throughput values for phases 1 and 2 were 50.0 and 55.7 seconds per mosquito, respectively, indicating a slight increase in time. In terms of effectiveness, the imager had 8% (6/80) critical errors and the loader had 13% (10/80) critical errors in phase 1. In phase 2, the imager (for VHT pairs) had 14% (11/80) critical errors and the loader (for VHT pairs) had 12% (19/160) critical errors. The average SUS score of the system was 70.25, indicating positive usability. A Kruskal-Wallis analysis demonstrated no significant difference in SUS (H value) scores between genders or users with and without smartphone use experience.
CONCLUSIONS: VectorCam is a usable system for deskilling the in-field identification of mosquito specimens in rural Uganda. Upcoming design updates will address the concerns of users and observers.

BACKGROUND: Vector control based on indoor residual spraying (IRS) is one of the main components of the visceral leishmaniasis (VL) elimination programme in India. Dichlorodiphenyltrichloroethane (DDT) was used for IRS until 2015 and was later replaced by the synthetic pyrethroid alpha-cypermethrin. Both classes of insecticides share the same target site, the voltage-gated sodium channel (Vgsc). As high levels of resistance to DDT have been documented in the local sand fly vector, Phlebotomus argentipes, it is possible that mutations in the Vgsc gene could provide resistance to alpha-cypermethrin, affecting current IRS pyrethroid-based vector control.
METHODS: This study aimed to compare frequencies of knockdown resistance (kdr) mutations in Vgsc between two sprayed and two unsprayed villages in Bihar state, India, which had the highest VL burden of the four endemic states. Across four villages, 350 female P. argentipes were collected as part of a 2019 molecular xenomonitoring study. DNA was extracted and used for sequence analysis of the IIS6 fragment of the Vgsc gene to assess the presence of kdr mutations.
RESULTS: Mutations were identified at various positions, most frequently at codon 1014, a common site known to be associated with insecticide resistance in mosquitoes and sand flies. Significant inter-village variation was observed, with sand flies from Dharampur, an unsprayed village, showing a significantly higher proportion of wild-type alleles (55.8%) compared with the three other villages (8.5-14.3%). The allele differences observed across the four villages may result from selection pressure caused by previous exposure to DDT.
CONCLUSIONS: While DDT resistance has been reported in Bihar, P. argentipes is still susceptible to pyrethroids. However, the presence of kdr mutations in sand flies could present a threat to IRS used for VL control in endemic villages in India. Continuous surveillance of vector bionomics and insecticide resistance, using bioassays and target genotyping, is required to inform India\'s vector control strategies and to ensure the VL elimination target is reached and sustained.

BACKGROUND: Soft ticks of the genus Ornithodoros are responsible for the maintenance and transmission of the African swine fever (ASF) virus in the sylvatic and domestic viral cycles in Southern Africa. They are also the main vectors of the Borrelia species causing relapsing fevers. Currently, no genetic markers are available for Afrotropical Ornithodoros ticks. As ASF spreads globally, such markers are needed to assess the role of ticks in the emergence of new outbreaks. The aim of this study is to design microsatellite markers that could be used for ticks of the Ornithodoros moubata complex, particularly Ornithodoros phacochoerus, to assess population structure and tick movements in ASF endemic areas.
METHODS: A total of 151 markers were designed using the O. moubata and O. porcinus genomes after elimination of repeated sequences in the genomes. All designed markers were tested on O. phacochoerus and O. porcinus DNA to select the best markers.
RESULTS: A total of 24 microsatellite markers were genotyped on two populations of O. phacochoerus and on individuals from four other Ornithodoros species. Nineteen markers were selected to be as robust as possible for population genetic studies on O. phacochoerus.
CONCLUSIONS: The microsatellite markers developed here represent the first genetic tool to study nidicolous populations of O. phacochoerus.

Diverse larval habitats significantly influence female mosquito oviposition. Utilizing traps that simulate these habitats is helpful in the study of the bioecology and characteristics of pathogen-transmitting species during oviposition. This study evaluated the feasibility of different traps in natural environments by comparing sampling methods and detecting the oviposition of epidemiologically important mosquitoes, with emphasis on Haemagogus species, in a fragment of the Atlantic Forest in Silva Jardim, Rio de Janeiro State, Brazil. Monthly collections were conducted from March 2021 to October 2023 using four types of traps: plastic containers, tires, bamboo, and sapucaia. Immatures were collected from these traps using a pipette, placed in plastic bags, and transported to the laboratory. Tire was the most efficient trap, showing the highest mosquito abundance (n = 1239) and number of species (S = 11). Conversely, the plastic container trap exhibited the lowest diversity (H = 0.43), with only two species and a low mosquito abundance (n = 26). The bamboo trap captured six species and recorded the second-highest diversity index (H = 1.04), while the sapucaia trap captured five species and had the third-highest diversity index (H = 0.91). Of the total immatures collected, 1817 reached adulthood, comprising 13 species, two of which are vectors of the sylvatic yellow fever virus: Haemagogus leucocelaenus and Haemagogus janthinomys. In conclusion, detecting key vectors of the sylvatic yellow fever virus in Brazil highlights the need for ongoing entomological and epidemiological surveillance in the study area and its vicinity. These efforts are crucial for monitoring vector presence and activity, identifying potential transmission hotspots, and devising effective control and prevention strategies.

BACKGROUND: Anopheles funestus is a leading vector of malaria in most parts of East and Southern Africa, yet its ecology and responses to vector control remain poorly understood compared with other vectors such as Anopheles gambiae and Anopheles arabiensis. This study presents the first large-scale survey of the genetic and phenotypic expression of insecticide resistance in An. funestus populations in Tanzania.
METHODS: We performed insecticide susceptibility bioassays on An. funestus mosquitoes in nine regions with moderate-to-high malaria prevalence in Tanzania, followed by genotyping for resistance-associated mutations (CYP6P9a, CYP6P9b, L119F-GSTe2) and structural variants (SV4.3 kb, SV6.5 kb). Generalized linear models were used to assess relationships between genetic markers and phenotypic resistance. An interactive R Shiny tool was created to visualize the data and support evidence-based interventions.
RESULTS: Pyrethroid resistance was universal but reversible by piperonyl-butoxide (PBO). However, carbamate resistance was observed in only five of the nine districts, and dichloro-diphenyl-trichloroethane (DDT) resistance was found only in the Kilombero valley, south-eastern Tanzania. Conversely, there was universal susceptibility to the organophosphate pirimiphos-methyl in all sites. Genetic markers of resistance had distinct geographical patterns, with CYP6P9a-R and CYP6P9b-R alleles, and the SV6.5 kb structural variant absent or undetectable in the north-west but prevalent in all other sites, while SV4.3 kb was prevalent in the north-western and western regions but absent elsewhere. Emergent L119F-GSTe2, associated with deltamethrin resistance, was detected in heterozygous form in districts bordering Mozambique, Malawi and the Democratic Republic of Congo. The resistance landscape was most complex in western Tanzania, in Tanganyika district, where all five genetic markers were detected. There was a notable south-to-north spread of resistance genes, especially CYP6P9a-R, though this appears to be interrupted, possibly by the Rift Valley.
CONCLUSIONS: This study underscores the need to expand resistance monitoring to include An. funestus alongside other vector species, and to screen for both the genetic and phenotypic signatures of resistance. The findings can be visualized online via an interactive user interface and could inform data-driven decision-making for resistance management and vector control. Since this was the first large-scale survey of resistance in Tanzania\'s An. funestus, we recommend regular updates with greater geographical and temporal coverage.

We present an annotated checklist of fleas (Siphonaptera) known to occur in the state of Delaware based on an examination of Siphonaptera collections at the University of Delaware and the Carnegie Museum of Natural History, as well as new specimens of fleas we collected from wildlife, other hosts, and tick flags. We review published records and compile them herein with our new records, which include 3 species previously unreported from Delaware. With these additions, there are now 18 flea species from 19 avian and mammalian hosts documented from Delaware.

BACKGROUND: Insecticide resistance (IR) is one of the major threats to malaria vector control programs in endemic countries. However, the mechanisms underlying IR are poorly understood. Thus, investigating gene expression patterns related to IR can offer important insights into the molecular basis of IR in mosquitoes. In this study, RNA-Seq was used to characterize gene expression in Anopheles gambiae surviving exposure to pyrethroids (deltamethrin, alphacypermethrin) and an organophosphate (pirimiphos-methyl).
RESULTS: Larvae of An. gambiae s.s. collected from Bassila and Djougou in Benin were reared to adulthood and phenotyped for IR using a modified CDC intensity bottle bioassay. The results showed that mosquitoes from Djougou were more resistant to pyrethroids (5X deltamethrin: 51.7% mortality; 2X alphacypermethrin: 47.4%) than Bassila (1X deltamethrin: 70.7%; 1X alphacypermethrin: 77.7%), while the latter were more resistant to pirimiphos-methyl (1.5X: 48.3% in Bassila and 1X: 21.5% in Djougou). RNA-seq was then conducted on resistant mosquitoes, non-exposed mosquitoes from the same locations and the laboratory-susceptible An. gambiae s.s. Kisumu strain. The results showed overexpression of detoxification genes, including cytochrome P450s (CYP12F2, CYP12F3, CYP4H15, CYP4H17, CYP6Z3, CYP9K1, CYP4G16, and CYP4D17), carboxylesterase genes (COEJHE5E, COE22933) and glutathione S-transferases (GSTE2 and GSTMS3) in all three resistant mosquito groups analyzed. Genes encoding cuticular proteins (CPR130, CPR10, CPR15, CPR16, CPR127, CPAP3-C, CPAP3-B, and CPR76) were also overexpressed in all the resistant groups, indicating their potential role in cross resistance in An. gambiae. Salivary gland protein genes related to \'salivary cysteine-rich peptide\' and \'salivary secreted mucin 3\' were also over-expressed and shared across all resistant groups.
CONCLUSIONS: Our results suggest that in addition to metabolic enzymes, cuticular and salivary gland proteins could play an important role in cross-resistance to multiple classes of insecticides in Benin. These genes warrant further investigation to validate their functional role in An. gambiae resistance to insecticides.

OBJECTIVE: To investigate the distribution of malaria vector Anopheles in Sichuan Province from 2011 to 2021, so as to provide the scientific evidence for improving the surveillance of malaria vector Anopheles and preventing re-establishment of imported malaria in Sichuan Province.
METHODS: The density and species of Anopheles mosquitoes were investigated using human-bait trapping and light trapping techniques in malaria vector surveillance sites of Sichuan Province from 2011 to 2021. The number, population and density of captured Anopheles mosquitoes were collected and descriptively analyzed, and the geographical distribution map of malaria vectors was plotted using the software ArcGIS 10.7 in Sichuan Province.
RESULTS: A total of 152 243 Anopheles mosquitoes were captured in malaria vector surveillance sites of Sichuan Province from 2011 to 2021, including 150 987 An. sinensis (99.18%) and 1 256 An. anthropophagus (0.82%), and no other Anopheles species were captured. The annual densities of An. sinensis and An. anthropophagus were 0.64 to 1.27 mosquitoes/(person-hour) and 0 to 0.07 mosquitoes/(person-hour) by the human-bait trapping technique, and 6.46 to 26.50 mosquitoes/(light-night) and 0 to 0.82 mosquitoes/(light-night) by the light trapping technique in malaria vector surveillance sites of Sichuan Province from 2011 to 2021. A relatively higher density of An. anthropophagus was seen in Renshou County, Jianyang City, Weiyuan County and Mabian Yi Autonomous County [> 0.40 mosquitoes/(person-hour)] by the human-bait trapping technique, and in Cuiping District and Gaoxian County in Yibin City [> 1.00 mosquito/(light-night)] by the light trapping technique in Sichuan Province from 2011 to 2018, with no An. anthropophagus captured from 2019 to 2021, and a relatively higher density of An. sinensis was detected in Emeishan City, Lushan County, Luojiang District, Tongchuan District and Zhaohua District [> 4.00 mosquitoes/(person-hour)] by the human-bait trapping technique, and in Huili County, Yuexi County, Dechang County, Langzhong City, Pingchang County and Xuanhan County [> 40.00 mosquitoes/(light-night)] by the light trapping technique in Sichuan Province from 2011 to 2021.
CONCLUSIONS: Malaria vectors were still widespread in Sichuan Province from 2011 to 2021, and An. sinensis was the dominant species of malaria vectors. There is still a risk of local re-establishment of imported malaria in Sichuan Province, and it is needed to continue to improve the surveillance of imported malaria cases and malaria vectors.
［摘要］ 目的 分析2011—2021年四川省传疟媒介按蚊分布, 为进一步提高传疟媒介按蚊监测质量、制定防止输入性 疟疾再传播措施提供科学依据。方法 2011—2021年, 于四川省传疟媒介监测点分别采用人诱法和灯诱法开展按蚊密 度及种群调查, 对按蚊捕获数量、种群及密度等进行描述性统计分析, 采用ArcGIS 10.7软件绘制四川省传疟媒介分布 图。结果 2011—2021年, 四川省传疟媒介监测点累计捕获按蚊152 243只, 其中中华按蚊150 987只 (占99.18%)、嗜人 按蚊1 256只 (占0.82%), 未捕获其他按蚊。2011—2021年, 四川省传疟媒介监测点采用人诱法调查发现, 中华按蚊年密 度为0.64 ~ 1.27只/(人·h)、嗜人按蚊年密度为0 ~ 0.07只/(人·h); 采用灯诱法调查发现, 中华按蚊年密度为6.46 ~ 26.50 只/(灯·夜)、嗜人按蚊年密度为0 ~ 0.82只/(灯·夜)。2011—2018年采用人诱法调查发现, 仁寿县、简阳市、威远县和马边 彝族自治县嗜人按蚊密度较高[> 0.40只/(人·h)]; 采用灯诱法调查发现, 宜宾市翠屏区和高县嗜人按蚊密度较高[> 1.00 只/(灯·夜)]; 2019—2021年未捕获嗜人按蚊。2011—2021年采用人诱法调查发现, 峨眉山市、芦山县、罗江区、通川区和 昭化区等5个监测点中华按蚊密度较高[均> 4.00只/(人·h)]; 采用灯诱法调查发现, 会理县、越西县、德昌县、阆中市、平 昌县和宣汉县等6个监测点中华按蚊密度较高[均> 40.00只/(灯·夜)]。结论 2011—2021年, 四川省传疟媒介依然广 泛分布, 中华按蚊为传疟优势按蚊; 输入性疟疾引起本地再传播风险依然存在, 需持续加强四川省输入性疟疾病例及传 疟媒介监测工作。.

We assessed the presence of Aedes aegypti in five ecorregions of Salta province and compared the oviposition activity of Ae. aegypti using ovitraps in towns of two contrasting ecoregions (yungas and Chaco dry forests) in the province of Salta, Argentina, a major contrast in these ecoregions are rain patterns and altitude. Our aim was to estimate how oviposition activities were associated with the ecoregion and site scale local environmental variables. Mosquito oviposition activity was monitored weekly during the summer using ovitraps. Predictor variables were ecoregion, town, and meteorological variables. The effect of the predictor variables was measured on the response variables using multi-model inference. Besides yungas, the presence of Aedes aegypti was confirmed in towns of dry Chaco and High Monte. The only factor that had a significant effect on the presence of eggs in the ovitraps was the ecoregion, with the frequency of positives being higher in yungas. For the number of eggs, the ecoregion, the night temperature of the first week and the NDVI would explain said variable. Overall, results indicate that the variations between towns would be more related with their ecological and climatic characteristics than with the more immediate meteorological variations.

Phlebotomine sand flies are crepuscular and nocturnal small dipteran insects in the family Psychodidae. Several disease agents, including Leishmania parasites, are transmitted to humans and other vertebrate hosts by the bite of an infected female sand fly. As part of leishmaniasis surveillance programs, light traps have been routinely used in sand fly collections. In this context, new trapping devices are always being required to improve vector monitoring. Here, the efficiency of a new suction light trap, named Silva suction trap or SS trap, was field evaluated in collecting sand flies. Two SS traps, one with green (520 nm, 15,000 mcd) and the other with white (wide spectrum, 18,000 mcd) LEDs, and one CDC-type trap were deployed in a rural forested environment. A total of 4686 phlebotomine sand flies were captured. The most frequent species were females of the Ps. Chagasi series (77.8%) followed by males of Ps. wellcomei (11.6%), Nyssomyia whitmani (3.3%), and Bichromomyia flaviscutellata (2.4%). The CDC-type light trap collected 101.9 ± 20.89 sand flies and 14 species, followed by the white-baited SS trap (87.78 ± 16.36, 14), and the green-baited SS trap (70.61 ± 14.75, 15), but there were no statistically significant differences among traps. A discussion on the considerable advantages of the use of SS traps over CDC traps is included. In this study, the Silva suction trap proved to be efficient and can be an alternative to CDC traps for monitoring adult phlebotomine sand fly populations.