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Climatic, ecological, and socioeconomic factors are facilitating the spread of mosquito-borne diseases, heightening the importance of vector surveillance and control. Citizen science is proving to be an effective tool to track mosquito populations, but methods are needed to detect and account for small scale sampling biases in citizen science surveillance. In this article we combine two types of traditional mosquito surveillance records with data from the Mosquito Alert citizen science system to explore the ways in which the socioeconomic characteristics of urban neighborhoods result in sampling biases in citizen scientists' mosquito reports, while also shaping the spatial distribution of mosquito populations themselves. We use Barcelona, Spain, as an example, and focus on Aedes albopictus, an invasive vector species of concern worldwide. Our results suggest citizen scientists' sampling effort is focused more in Barcelona's lower and middle income census tracts than in its higher income ones, whereas Ae. albopictus populations are concentrated in the city's upper-middle income tracts. High resolution estimates of the spatial distribution of Ae. albopictus risk can be improved by controlling for citizen scientists' sampling effort, making it possible to provide better insights for efficiently targeting control efforts. Our methodology can be replicated in other cities faced with vector mosquitoes to improve public health responses to mosquito-borne diseases, which impose massive burdens on communities worldwide. [ABSTRACT FROM AUTHOR]

Vesicular stomatitis (VS) is a viral disease that affects horses, cattle, and swine that is transmitted by direct contact and hematophagous insects. In 2023, a multi-state outbreak of vesicular stomatitis New Jersey virus (VSNJV) occurred in California, Nevada, and Texas, infecting horses, cattle, and rhinoceros. To identify possible insect vectors, we conducted insect surveillance at various locations in San Diego County, CA, including at a wildlife park. CO2 baited traps set from mid-May to mid-August 2023 collected 2357 Culicoides biting midges and 1215 Simulium black flies, which are insect genera implicated in VSNJV transmission. Insects were pooled by species, location, and date, then tested for viral RNA. Nine RNA-positive pools of Culicoides spp. and sixteen RNA-positive pools of Simulium spp were detected. Infectious virus was detected by cytopathic effect in 96% of the RNA-positive pools. This is the first report of VSNJV in wild-caught C. bergi, C. freeborni, C. occidentalis, S. argus, S. hippovorum, and S. tescorum. The vector competency of these species for VSNJV has yet to be determined but warrants examination. Active vector surveillance and testing during disease outbreaks increases our understanding of the ecology and epidemiology of VS and informs vector control efforts. [ABSTRACT FROM AUTHOR]

Abstract Climatic, ecological, and socioeconomic factors are facilitating the spread of mosquito-borne diseases, heightening the importance of vector surveillance and control. Citizen science is proving to be an effective tool to track mosquito populations, but methods are needed to detect and account for small scale sampling biases in citizen science surveillance. In this article we combine two types of traditional mosquito surveillance records with data from the Mosquito Alert citizen science system to explore the ways in which the socioeconomic characteristics of urban neighborhoods result in sampling biases in citizen scientists’ mosquito reports, while also shaping the spatial distribution of mosquito populations themselves. We use Barcelona, Spain, as an example, and focus on Aedes albopictus, an invasive vector species of concern worldwide. Our results suggest citizen scientists’ sampling effort is focused more in Barcelona’s lower and middle income census tracts than in its higher income ones, whereas Ae. albopictus populations are concentrated in the city’s upper-middle income tracts. High resolution estimates of the spatial distribution of Ae. albopictus risk can be improved by controlling for citizen scientists’ sampling effort, making it possible to provide better insights for efficiently targeting control efforts. Our methodology can be replicated in other cities faced with vector mosquitoes to improve public health responses to mosquito-borne diseases, which impose massive burdens on communities worldwide.

Mosquitoes (Diptera: Culicidae) and the pathogens they transmit represent a threat to human and animal health. Low-cost and effective surveillance methods are necessary to enable sustainable monitoring of mosquito distributions, diversity, and human interactions. This study examined the use of iNaturalist, an online, community-populated biodiversity recording database, for passive mosquito surveillance in the United Kingdom (UK) and Ireland, countries under threat from the introduction of invasive mosquitoes and emerging mosquito-borne diseases. The Mozzie Monitors UK & Ireland iNaturalist project was established to collate mosquito observations in these countries. Data were compared with existing long-term mosquito UK datasets to assess representativeness of seasonal and distribution trends in citizen scientist-recorded observations. The project collected 738 observations with the majority recorded 2020-2022. Records were primarily associated with urban areas, with the most common species Culex pipiens and Culiseta annulata significantly more likely to be observed in urban areas than other species. Analysis of images uploaded to the iNaturalist project also provided insights into human-biting behavior. Our analyses indicate that iNaturalist provides species composition, seasonal occurrence, and distribution figures consistent with existing datasets and is therefore a useful surveillance tool for recording information on human interactions with mosquitoes and monitoring species of concern. [ABSTRACT FROM AUTHOR]

Abstract 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. Graphical Abstract

Abstract 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. Graphical Abstract

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Abstract 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. Graphical Abstract

Abstract 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. Conclusion 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.

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. [ABSTRACT FROM AUTHOR]

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. Conclusion: 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. [ABSTRACT FROM AUTHOR]

Objective To explore the application of time series models based on meteorological factors in the population density of Aedes albopictus in Hainan Province, and to provide a scientific basis for the prevention and control of dengue fever in Hainan Province. Methods The density of Aedes albopictus in different habitats in 18 cities and counties of Hainan Province from 2017 to 2022 was monitored monthly using the double-mosquito net trapping index and the Breteau index. Mann-Kendall trend test was used to analyze the temporal trends of the two density indices; Spearman's correlation analysis was employed to assess the correlation strength between each meteorological factor and the two indexes, eliminating unrelated variables, and further selecting the final variables through the full-subset regression method. Three time-series models were constructed for the two density indices, with root mean square error (RMSE), mean absolute error (MAE), and other accuracy metrics used to determine the optimal model; predictions for the density indices for 2023-2024 were made. All statistical analyses were performed in R (4.3.1). Results The net trapping and Breteau indices showed an overall decrease over the years (Z-values of Mann-Kendall trend test were -6.15 and -4.03, respectively, and P<0.05). The meteorological factor most strongly associated with the trap index was the monthly average minimum temperature; monthly mean minimum temperature and monthly mean relative humidity were strongly correlated with the Breteau index. Based on various evaluation indicators, the multivariate time series model demonstrated the highest accuracy. The study predicts one to two peaks in both the trap index and Breteau Index for the years 2023 and 2024, with peak periods occurring between June to September and May to September, respectively. The predicted value for 2023 aligns with the measured value, demonstrating outstanding predictive accuracy. Conclusions This study has introduced meteorological factors into the seasonal time series model, allowing for more accurate predictions of the density of Aedes albopictus in Hainan Province from 2023 to 2024, providing a model framework for the prevention and control of dengue fever in Hainan Province. [ABSTRACT FROM AUTHOR]

Background & objectives: Dengue is a major vector-borne disease having public health importance. It is caused by Dengue Virus (DENV) and is transmitted by mosquitoes of Aedes species. With the unavailability of a vaccine, vector control remains the only preventive measure for dengue. Studies have already been conducted to establish the presence of dengue vectors in the north-eastern states of India. However, limited studies have been conducted in Tripura state. In the present study we aimed to identify the preferred breeding habitats of dengue vectors in the state. Methods: Clinical case data of dengue since the last five years was studied and the areas with the highest case numbers were identified. Entomological investigation was carried out in areas reporting the highest number of cases. Larvae were collected from the breeding habitats using standard protocol followed by morphological and molecular identification. Further, House index (HI), Container index (CI) and Pupal index (PI) were determined. The positive pools were then processed for incrimination for the presence of dengue virus. Calculation of entomological indices was done. Results: Of the total 815 containers searched, 36.80% containers were positive for mosquito larvae. Among the immature mosquito collection, 836 adults emerged and were identified as Aedes albopictus using standard taxonomic keys followed by molecular methods. HI, CI and PI, varied from 15.38% to 100%, 21% to 31.04 %, and 2.93% to 110.53% respectively. However, none of the pools was positive for dengue virus. Interpretation & conclusion: The present study identified Ae. albopictus as a potential vector of dengue in Tripura. The study gave important insights on the preferred larval habitats and provides information on the indication of displacement of Ae. albopictus from rural to urban and semi-urban areas. However, longitudinal studies for longer time frame are necessary for any conclusive remarks.

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. [ABSTRACT FROM AUTHOR]

Since 2015, the Dengue, Zika, and Chikungunya viruses gained notoriety for their impact in public health in many parts of the globe, including Brazil. In Recife, the capital of Pernambuco State, the introduction of ZIKV impacted human population tremendously, owing to the increase in the number of neurological cases, such as the Guillain−Barré and congenital Zika disorders. Later, Recife was considered to be the epicenter for ZIKV epidemics in Brazil. For arboviral diseases, there are some risk factors, such as climate changes, low socioeconomic conditions, and the high densities of vectors populations, that favor the broad and rapid dispersion of these three viruses in the city. Therefore, continuous arbovirus surveillance provides an important tool for detecting these arboviruses and predicting new outbreaks. The purpose of the present study was to evaluate the circulation of DENV, ZIKV, and CHIKV by RT-qPCR in mosquitoes collected in health care units from the metropolitan area of Recife (MAR), during 2018. A total of 2321 female mosquitoes (357 pools) belonging to two species, Aedes aegypti and Culex quinquefasciatus, were collected from 18 different healthcare units, distributed in five cities from the MAR. Twenty-three pools were positive for ZIKV, out of which, seventeen were of C. quinquefasciatus and six were of A. aegypti. Positive pools were collected in 11/18 health care units screened, with Cq values ranging from 30.0 to 37.4 and viral loads varying from 1.88 × 107 to 2.14 × 109 RNA copies/mL. Nosocomial Aedes- and Culex-borne transmission of arbovirus are widely ignored by surveillance and vector control programs, even though healthcare-associated infections (HAI) are considered a serious threat to patient safety worldwide. Although the results presented here concern only the epidemiological scenario from 2018 in MAR, the potential of hospital-acquired transmission through mosquito bites is being overlooked by public health authorities. It is, therefore, of the ultimate importance to establish specific control programs for these locations.

Abstract Background Anopheles stephensi is an efficient vector of both Plasmodium falciparum and Plasmodium vivax in South Asia and the Middle East. The spread of An. stephensi to countries within the Horn of Africa threatens progress in malaria control in this region as well as the rest of sub-Saharan Africa. Methods The available malaria data and the timeline for the detection of An. stephensi was reviewed to analyse the role of An. stephensi in malaria transmission in Horn of Africa of the Eastern Mediterranean Region (EMR) in Djibouti, Somalia, Sudan and Yemen. Results Malaria incidence in Horn of Africa of EMR and Yemen, increased from 41.6 in 2015 to 61.5 cases per 1000 in 2020. The four countries from this region, Djibouti, Somalia, Sudan and Yemen had reported the detection of An. stephensi as of 2021. In Djibouti City, following its detection in 2012, the estimated incidence increased from 2.5 cases per 1000 in 2013 to 97.6 cases per 1000 in 2020. However, its contribution to malaria transmission in other major cities and in other countries, is unclear because of other factors, quality of the urban malaria data, human mobility, uncertainty about the actual arrival time of An. stephensi and poor entomological surveillance. Conclusions While An. stephensi may explain a resurgence of malaria in Djibouti, further investigations are needed to understand its interpretation trends in urban malaria across the greater region. More investment for multisectoral approach and integrated surveillance and control should target all vectors particularly malaria and dengue vectors to guide interventions in urban areas.

Vector surveillance is crucial for implementation of appropriate vector control strategies to reduce the transmission of vector-borne diseases such as dengue, a major public health concern in Sri Lanka. Aedes aegypti and Ae. albopictus are the primary and secondary vectors of dengue virus, respectively. Dengue is endemic in the Jaffna District and the presence of dengue vectors from many water bodies, including brackish water habitats, is reported. The use of environmental DNA (eDNA) has emerged as a novel tool to detect and identify vector species present in any aquatic environment. This study, used environmental DNA to determine the presence of major dengue vectors in different aquatic habitats in the Municipal limits of the Jaffna District. Potential pre-imaginal developmental sites of dengue vectors were identified in randomly selected 10 blocks consisting of at least 40 houses within the study location. One hundred and twenty water samples without any visible preimaginal forms were collected from these sites to extract eDNA. PCR assays were performed using species-specific primers designed for Ae. aegypti and Ae. albopictus. The presence of Ae. aegypti and Ae. albopictus was identified in 12 (10.0%) and 30 samples (25.0%), respectively. The presence of both species was detected in nine samples (7.5%). The salinity of the water from which Aedes DNA was detected ranged from 0–7 gL-1. The results revealed that eDNA is an indirect but valid surveillance tool for monitoring preimaginal developmental forms of mosquitos in the aquatic environment.

Ovitraps can detect Aedes vectors at an early stage and can serve as an alarm indicator for outbreak prediction. This study aimed to summarize the available literature about the ovitrap system and to determine its feasibility, required resources and costs when installing and maintaining this vector surveillance system in the municipality of Los Patios, Colombia. A scoping review to assess the role of ovitraps as a tool for Aedes vector surveillance was conducted. The subsequent fieldwork consisted of mapping the municipality, manufacturing, and installing 40 ovitraps in 10 blocks, revising them weekly for 4 weeks by two half-time employed vector control technicians, and carrying out a cost analysis. A total of 38 studies were included in this review showing that ovitraps had a better performance than other entomological surveillance methods and a positive correlation with other entomological and disease variables. From the field results over 4 weeks, a high proportion of positive ovitraps (80%, 90%, 75%, 97.5%) and positive blocks (100%) as well as a good acceptance by house owners (76.9%), were identified. Operational indicators such as average installation time of the ovitraps (10h15 m), weekly reading and reinstallation (on average 7h27 m) and the cost of the intervention (COL$1,142,304.47/US$297) were calculated. Literature shows that ovitraps are sensitive to detect the presence of Aedes mosquitoes, providing data efficiently and timely for outbreak prediction. The field testing showed it is an affordable and feasible method in the context of a Colombian municipality and similar endemic areas. [ABSTRACT FROM AUTHOR]

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.

Anopheles mosquitoes present a major public health challenge in sub-Saharan Africa; notably, as vectors of malaria that kill over half a million people annually. In parts of the east and southern Africa region, one species in the Funestus group, Anopheles funestus, has established itself as an exceptionally dominant vector in some areas, it is responsible for more than 90% of all malaria transmission events. However, compared to other malaria vectors, the species is far less studied, partly due to difficulties in laboratory colonization and the unresolved aspects of its taxonomy and systematics. Control of An. funestus is also increasingly difficult because it has developed widespread resistance to public health insecticides. Fortunately, recent advances in molecular techniques are enabling greater insights into species identity, gene flow patterns, population structure, and the spread of resistance in mosquitoes. These advances and their potential applications are reviewed with a focus on four research themes relevant to the biology and control of An. funestus in Africa, namely: (i) the taxonomic characterization of different vector species within the Funestus group and their role in malaria transmission; (ii) insecticide resistance profile; (iii) population genetic diversity and gene flow, and (iv) applications of genetic technologies for surveillance and control. The research gaps and opportunities identified in this review will provide a basis for improving the surveillance and control of An. funestus and malaria transmission in Africa. [ABSTRACT FROM AUTHOR]

Background: Anopheles stephensi is an efficient vector of both Plasmodium falciparum and Plasmodium vivax in South Asia and the Middle East. The spread of An. stephensi to countries within the Horn of Africa threatens progress in malaria control in this region as well as the rest of sub-Saharan Africa. Methods: The available malaria data and the timeline for the detection of An. stephensi was reviewed to analyse the role of An. stephensi in malaria transmission in Horn of Africa of the Eastern Mediterranean Region (EMR) in Djibouti, Somalia, Sudan and Yemen. Results: Malaria incidence in Horn of Africa of EMR and Yemen, increased from 41.6 in 2015 to 61.5 cases per 1000 in 2020. The four countries from this region, Djibouti, Somalia, Sudan and Yemen had reported the detection of An. stephensi as of 2021. In Djibouti City, following its detection in 2012, the estimated incidence increased from 2.5 cases per 1000 in 2013 to 97.6 cases per 1000 in 2020. However, its contribution to malaria transmission in other major cities and in other countries, is unclear because of other factors, quality of the urban malaria data, human mobility, uncertainty about the actual arrival time of An. stephensi and poor entomological surveillance. Conclusions: While An. stephensi may explain a resurgence of malaria in Djibouti, further investigations are needed to understand its interpretation trends in urban malaria across the greater region. More investment for multisectoral approach and integrated surveillance and control should target all vectors particularly malaria and dengue vectors to guide interventions in urban areas. [ABSTRACT FROM AUTHOR]

Despite the annual implementation of a robust and extensive indoor residual spraying programme against malaria vectors in Limpopo Province (South Africa), significant transmission continues and is a serious impediment to South Africa's malaria elimination objectives. In order to gain a better understanding regarding possible causes of this residual malaria, we conducted a literature review of the historical species composition and abundance of malaria vector mosquitoes in the Limpopo River Valley region of the Vhembe District, northern Limpopo Province, the region with the highest remaining annual malaria cases in South Africa. In addition, mosquito surveys were carried out in the same region between October 2017 and October 2018. A total of 2225 adult mosquitoes were collected using CO2-baited tent and light traps, human landing catches and cow-baited traps. Of the 1443 Anopheles collected, 516 were members of the An. gambiae complex and 511 An. funestus group. In the malaria endemic rural areas outside the Kruger National Park, one specimen each of An. gambiae s.s. and An. funestus and only three of An. arabiensis were collected. The latter species was abundant at a remote hot spring in the neighboring Kruger National Park. Eighteen other species of Anopheles were collected. Our survey results support the historical findings that An. arabiensis, the species widely held to be the prime malaria vector in South Africa, is a rare species in the malaria endemic Limpopo River Valley. The implications of the mosquito surveys for malaria transmission, elimination and vector control in northern Limpopo Province and neighboring regions are discussed.

BackgroundSurveillance of outdoor resting malaria vector populations is crucial to monitor possible changes in vector resting and feeding behaviour following the widespread use of indoor-based vector control interventions. However, it is seldom included in the routine vector surveillance system in Africa due to lack of well standardized and efficient traps. This study was conducted to evaluate the performance of sticky pots for outdoor resting malaria vector surveillance in western Kenya.MethodsMosquito collections were conducted from September 2015 to April 2016 in Ahero and Iguhu sites, western Kenya using sticky pots, pit shelters, clay pots, exit traps, Prokopack aspirator and CDC light traps (outdoor and indoor). Species within Anopheles gambiae (s.l.) were identified using polymerase chain reaction (PCR). Enzyme-linked immunosorbent assay (ELISA) was used to determine blood meal sources of malaria vectors.ResultsA total of 23,772 mosquitoes were collected, of which 13,054 were female anophelines comprising An. gambiae (s.l.) (72.9%), An. funestus (13.2%), An. coustani (8.0%) and An. pharoensis (5.9%). Based on PCR assay (n = 672), 98.6% An. arabiensis and 1.4% An. gambiae (s.s.) constituted An. gambiae (s.l.) in Ahero, while this was 87.2% An. gambiae (s.s.) and 12.8% An. arabiensis in Iguhu. The sticky pots and pit shelters showed similar performance with regard to the relative abundance and host blood meal indices of An. gambiae (s.l.) and An. funestus. In terms of density per trap, a pit shelter caught on average 4.02 (95% CI: 3.06-5.27) times as many An. gambiae (s.l.) as a sticky pot, while a sticky pot captured 1.60 (95% CI: 1.19-2.12) times as many An. gambiae (s.l.) as a clay pot. Exit traps yielded a significantly lower number of An. gambiae (s.l.) than all other traps in Ahero, but a higher number of An. gambiae (s.l.) compared to the other outdoor traps in Iguhu. Indoor CDC light traps captured a significantly higher number of An. funestus than other traps.ConclusionsSticky pots could be a useful and complementary tool for outdoor resting malaria vector surveillance, in settings where using pit shelters is not feasible and less productive. The lower vector density in the sticky pots compared to pit shelters suggests that batches of sticky pots (i.e. four per compound) need to be deployed in order to make a direct comparison. This study also highlighted the need to concurrently undertake indoor and outdoor vector surveillance to better understand residual malaria transmission.

BACKGROUND:Dengue is the most prevalent mosquito-borne disease in the world, with China affected seriously in recent years. 65.8% of dengue cases identified in mainland China since 2005 were reported from the city of Guangzhou. METHODS:In this study, we described the incidence rate and distribution of dengue cases using data collected form National Notifiable Infectious Disease Reporting Information System data in Guangzhou for 2001 to 2016. All dengue cases were investigated using standardized questionnaire. RESULTS:A total of 42,469 dengue cases were reported, with an average annual incidence rate of 20.99 per 100,000 resident population. Over this time period, the incidence rate of indigenous cases increased. Dengue affected areas also expanded sharply geographically from 58.1% of communities affected during 2001-2005 to 96.4% of communities affected in 2011-2016. Overall 95.30% of the overseas imported cases were reported during March and December, while 99.79% of indigenous cases were reported during July and November. All four dengue virus serotypes were identified both in imported cases and indigenous cases. The Aedes albopictus mosquito was the only vector for dengue transmission in the area. CONCLUSIONS:Guangzhou has become the dengue epicenter in mainland China. Control strategies for dengue should be adjusted to the epidemiological characteristics above and intensive study need to be conducted to explore the factors that driving the rapid increase of dengue.

Abstract Background Aedes albopictus is a highly invasive species and an important vector of dengue and chikungunya viruses. Indigenous to Southeast Asia, Ae. albopictus has successfully invaded every inhabited continent, except Antarctica, in the past 80 years. Vector surveillance and control at points of entry (PoE) is the most critical front line of defence against the introduction of Ae. albopictus to new areas. Identifying the pathways by which Ae. albopictus are introduced is the key to implementing effective vector surveillance to rapidly detect introductions and to eliminate them. Methods A literature review was conducted to identify studies and data sources reporting the known and suspected dispersal pathways of human-mediated Ae. albopictus dispersal between 1940–2020. Studies and data sources reporting the first introduction of Ae. albopictus in a new country were selected for data extraction and analyses. Results Between 1940–2020, Ae. albopictus was reported via various dispersal pathways into 86 new countries. Two main dispersal pathways were identified: (1) at global and continental spatial scales, maritime sea transport was the main dispersal pathway for Ae. albopictus into new countries in the middle to late 20th Century, with ships carrying used tyres of particular importance during the 1980s and 1990s, and (2) at continental and national spatial scales, the passive transportation of Ae. albopictus in ground vehicles and to a lesser extent the trade of used tyres and maritime sea transport appear to be the major drivers of Ae. albopictus dispersal into new countries, especially in Europe. Finally, the dispersal pathways for the introduction and spread of Ae. albopictus in numerous countries remains unknown, especially from the 1990s onwards. Conclusions This review identified the main known and suspected dispersal pathways of human-mediated Ae. albopictus dispersal leading to the first introduction of Ae. albopictus into new countries and highlighted gaps in our understanding of Ae. albopictus dispersal pathways. Relevant advances in vector surveillance and genomic tracking techniques are presented and discussed in the context of improving vector surveillance. Graphical Abstract

Abstract There are increasing reports of the Asian malaria mosquito, Anopheles stephensi invading and spreading in Eastern Africa. We discuss the importance of these invasions in the context of broader challenges facing malaria control in Africa and argue against addressing it as an isolated problem. Anopheles stephensi is only one of multiple biological threats facing malaria control in the region—and is itself an indication of wide-ranging weaknesses in vector surveillance and control programs. Expanded investigations are needed in both urban and rural areas, especially in countries serviced by the Indian Ocean trade routes, to establish the full extent and future trajectories of the problem. More importantly, instead of tackling this vector species as a stand-alone threat, affected countries should adopt more integrated and multi-sectorial initiatives that can sustainably drive and keep out malaria.

Abstract Background Proper vector surveillance relies on the ability to identify species of interest accurately and efficiently, though this can be difficult in groups containing cryptic species. Culicoides Latreille is a genus of small biting flies responsible for the transmission of numerous pathogens to a multitude of vertebrates. Regarding pathogen transmission, the C. variipennis species complex is of particular interest in North America. Of the six species within this group, only C. sonorensis Wirth & Jones is a proven vector of bluetongue virus and epizootic hemorrhagic disease virus. Unfortunately, subtle morphological differences, cryptic species, and mitonuclear discordance make species identification in the C. variipennis complex challenging. Recently, single-nucleotide polymorphism (SNP) analysis enabled discrimination between the species of this group; however, this demanding approach is not practical for vector surveillance. Methods The aim of the current study was to develop a reliable and affordable way of distinguishing between the species within the C. variipennis complex, especially C. sonorensis. Twenty-five putative microsatellite markers were identified using the C. sonorensis genome and tested for amplification within five species of the C. variipennis complex. Machine learning was then used to determine which markers best explain the genetic differentiation between species. This led to the development of a subset of four and seven markers, which were also tested for species differentiation. Results A total of 21 microsatellite markers were successfully amplified in the species tested. Clustering analyses of all of these markers recovered the same species-level identification as the previous SNP data. Additionally, the subset of seven markers was equally capable of accurately distinguishing between the members of the C. variipennis complex as the 21 microsatellite markers. Finally, one microsatellite marker (C508) was found to be species-specific, only amplifying in the vector species C. sonorensis among the samples tested. Conclusions These microsatellites provide an affordable way to distinguish between the sibling species of the C. variipennis complex and could lead to a better understanding of the species dynamics within this group. Additionally, after further testing, marker C508 may allow for the identification of C. sonorensis with a single-tube assay, potentially providing a powerful new tool for vector surveillance in North America. Graphical Abstract

Kentucky experiences some of the highest incidence rates for ehrlichiosis nationwide. Ehrlichiosis is a bacterial infection caused primarily by the pathogen Ehrlichia chaffeensis and can be transmitted to humans through the bite of an infected tick, notably Amblyomma americanum. Amblyomma americanum, the lone star tick, is common to Kentucky and much of the southeast, but has expanded farther north in recent years. As an abundant and aggressive nondiscriminatory biter, this species is of major public health concern for transmission of pathogens to humans. As this vector's range expands, surveillance remains a necessary tool providing data that allows researchers to track this expansion over time. The historical information on tick distribution in Kentucky is variable with very little data on a statewide scale. From January 2019 to December 2020, we conducted surveillance for A. americanum in Kentucky through field collections and the establishment of a statewide tick submission program with the help of the Kentucky Department for Public Health and screened for E. chaffeensis on a county-level throughout the state. We collected 5,726 A. americanum ticks in 77 counties and detected E. chaffeensis in 32 counties. The minimum infection rate was 1.8%. With the expansion of A. americanum and increasing cases of tick-borne diseases, future surveillance is needed to monitor this important tick vector over time.

Objectives: Following the Public Health Emergency of International Concern declared on Zika by the World Health Organization during 2016, the Indian Council of Medical Research carried out nationwide vector surveillance for Zika and Dengue viruses (ZIKV and DENV) in India as a preparedness measure in 2016–19. Methods: High-risk zones distributed to 49 Districts in 14 states/union territories were included in the study. Seven ICMR institutions participated, following a standard operating protocol. Aedes specimens sampled weekly were processed by multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) for ZIKV/DENV and random samples crosschecked with real-time RT-PCR for ZIKV. Results: Altogether, 79 492 Aedes specimens in 6492 pools were processed; 3 (0.05%) and 63 (0.97%) pools, respectively, were found positive for ZIKV and DENV. ZIKV infections were recorded in Aedes aegypti sampled during the 2018 sporadic Zika outbreak in Jaipur, Rajasthan. However, these belonged to the Asian lineage of the virus, already circulating in the country. Both Ae. aegypti and Aedes albopictus distributed to 8 states/union territories were found to be infected with DENV. Both sexes of Ae. albopictus were infected, indicating transovarial transmission. Conclusion: This investigation evinced no active transmission of the American lineage-pandemic Zika virus in India during the pandemic period.

Tick abundance and diagnosed cases of tick-borne diseases have been increasing in the United States. American black bear (Ursus americanus) populations have also been increasing in the eastern United States. As a competent host of several species of ticks and a mammal capable of traveling long distances, the role of black bears as hosts for ticks requires further evaluation. Ectoparasite surveys were conducted on black bears in Pennsylvania to evaluate tick presence, abundance, spatial distribution, and association with Sarcoptes scabiei, the etiological agent of sarcoptic mange, on bears to better understand their role in tick ecology and to improve on-host surveillance techniques. Tick burden was evaluated using standard area sampling (10.16 × 10.16 cm squares) on pre-designated body regions on black bears from June 2018–December 2019. In total, 278 unique individual black bears were evaluated, with all ticks identified as Ixodes scapularis (n = 1976; 76.7% adults, 23.3% immatures). Tick presence differed by body region on bears, with the highest percentage of tick observations located on bear ears and muzzle. Ticks also partitioned on black bears by life-stage, with immature ticks primarily recorded on the lower extremities of bears and adult ticks primarily recorded on the front-quarters of bears. This includes the first known record of I. scapularis larvae parasitizing black bears, and observations of all three mobile life-stages concurrently parasitizing bears. Tick abundance was also statistically significant dependent on season, with the highest abundance of ticks recorded in spring and lowest abundance in fall. Adult ticks were less likely to be present on bears with mange. These data reveal the important role black bears may serve in tick ecology and dispersal as all three mobile life-stages of I. scapularis were found parasitizing a mammal capable of traveling far distances in a region with high numbers of Lyme disease cases.

Objective: To determine the effect of comprehensive campaign for dengue control in Southern Punjab, Pakistan. Study Design: Cross sectional study. Place and Duration of the Study: Station Health Organization Multan and Departments of Pathology, Combined Military Hospital Multan, from Mar 2019 to Dec 2020. Methodology: This study was conducted by implementing a comprehensive campaign by involvement of public health workers in high risk dengue area of Multan. There were five phases of this campaign; awareness, situation analysis, vector surveillance, indoor residual sprays where vector was found and contact tracing of dengue fever cases. The system was developed in 46 Mohallas/localities of Multan. Hot spot for mosquitoes breeding were identified at 15 places. Total of 23 health workers were recruited including 3 female workers. Insecticide spraying was carried out by indoor residual spray pumps & thermal foggers in a cyclic pattern. Results: During 2019, 42 larvae were identified followed by insecticide sprays & follow up visits; 26 cases of dengue fever were diagnosed and treated at CMH Multan in a dedicated dengue ward. Contact tracing was carried out and indoor residual sprays were carried out in 20 houses around residence of these cases. In 2020, 5 larvae were identified followed by sprays & follow up visits; 2 cases of dengue fever were diagnosed and treated. Conclusion: Comprehensive campaign against dengue fever was quite successful to control spread of dengue fever.

Background Dengue virus (DENV) and Zika virus (ZIKV) have a severe impact on human health worldwide. To understand the dynamics of these viruses in mosquito populations, it is necessary to maintain surveillance during non-epidemic years. Methods We aimed to assess the presence of DENV and ZIKV in Aedes aegypti females in the state of Sergipe, northeastern Brazil, during a non-epidemic year. The Ae. aegypti females collected were sectioned, and the heads and thorax were used to analyze the infection rate. Each female was first analyzed to detect Flavivirus using RT-PCR. Flavivirus-positive samples were further screened to detect ZIKV and DENV types 1, 2, 3 and 4. Results A total of 184 females were collected. ZIKV was identified in 5.4% of the sample and DENV in 7.1%, with the DENV-positive samples belonging to subtypes 2, 3 and 4. The presence of coinfected vectors was also observed. Of the four cities tested, only one was negative for all viruses. Conclusions These results show that the maintenance of vigilance during non-epidemic years can provide data on viruses circulating before the onset of outbreaks. This can enable the planning and implementation of local control measures to prevent the appearance of new outbreaks. [ABSTRACT FROM AUTHOR]

Mosquito-borne diseases continue to pose a great threat to global public health systems due to increased insecticide resistance and climate change. Accurate vector identification is crucial for effective control, yet it presents significant challenges. IDX - an automated computer vision-based device capable of capturing mosquito images and outputting mosquito species ID has been deployed globally resulting in algorithms currently capable of identifying 53 mosquito species. In this study, we evaluate deployed performance of the IDX mosquito species identification algorithms using data from partners in the Southeastern United States (SE US) and Papua New Guinea (PNG) in 2023 and 2024. This preliminary assessment indicates continued improvement of the IDX mosquito species identification algorithms over the study period for individual species as well as average regional accuracy with macro average recall improving from 55.3 % [Confidence Interval (CI) 48.9, 61.7] to 80.2 % [CI 77.3, 84.9] for SE US, and 84.1 % [CI 75.1, 93.1] to 93.6 % [CI 91.6, 95.6] for PNG using a CI of 90 %. This study underscores the importance of algorithm refinement and dataset expansion covering more species and regions to enhance identification systems thereby reducing the workload for human experts, addressing taxonomic expertise gaps, and improving vector control efforts., Competing Interests: Declaration of competing interest All authors are employed by and partial owners of Vectech, the company developing and distributing the IDX hardware and software, (Copyright © 2024. Published by Elsevier B.V.)

The Arizona State University (ASU) Tempe campus is inhabited by some 55,000 enrolled students as well as several mosquito species that can transmit West Nile virus, dengue, Zika, chikungunya, and yellow fever. The time of host-seeking of these vectors has not been quantified on the ASU Tempe campus, but this information is important to inform ground and/or truck-mount fogging operations targeting mosquitoes to prevent or control disease outbreaks. We quantified the time of host-seeking of the predominant mosquito vector species at the ASU Tempe campus during the post-monsoon season in 2021, using collection bottle rotators with encephalitis vector survey traps that were baited with CO2, at 3 h intervals during a full day. Culex quinquefasciatus, Aedes aegypti, and Culex tarsalis were the most abundant species captured. Pre-midnight host-seeking (18:00-00:00) accounted for 52% of all captures, whereas post-midnight host-seeking (00:00-06:00) accounted for 35% of all captures. Peak activity times were between 21:00 and 00:00 for Cx. quinquefasciatus and Cx. tarsalis, and between 15:00 and 18:00 for Ae. aegypti. Data can be used to inform local mosquito surveillance and control programs., (Copyright © 2024 by The American Mosquito Control Association, Inc.)

Abstract Background Triatomine bugs transmit Chagas disease across Latin America, where vector control-surveillance is increasingly decentralized. Locally run systems often deal with highly diverse native-vector faunas—plus, in some areas, domestic populations of non-native species. Flexible entomological-risk indicators that cover native and non-native vectors and can support local decision-making are therefore needed. Methods We present a local-scale entomological-risk score (“TriatoScore”) that leverages and builds upon information on the ecology-behavior and distribution-biogeography of individual triatomine bug species. We illustrate our approach by calculating TriatoScores for the 417 municipalities of Bahia state, Brazil. For this, we (i) listed all triatomine bug species recorded statewide; (ii) derived a “species relevance score” reflecting whether each species is native/non-native and, if native, whether/how often it invades/colonizes dwellings; (iii) mapped each species’ presence by municipality; (iv) for native vectors, weighted presence by the proportion of municipal territory within ecoregions occupied by each species; (v) multiplied “species relevance score” × “weighted presence” to get species-specific “weighted scores”; and (vi) summed “weighted scores” across species to get municipal TriatoScores. Using standardized TriatoScores, we then grouped municipalities into high/moderate/low entomological-risk strata. Results TriatoScores were higher in municipalities dominated by dry-to-semiarid ecoregions than in those dominated by savanna-grassland or, especially, moist-forest ecoregions. Bahia’s native triatomines can maintain high to moderate risk of vector-borne Chagas disease in 318 (76.3%) municipalities. Historical elimination of Triatoma infestans from 125 municipalities reduced TriatoScores by ~ 27% (range, 20–44%); eight municipalities reported T. infestans since Bahia was certified free of Trypanosoma cruzi transmission by this non-native species. Entomological-risk strata based on TriatoScores agreed well with Bahia’s official disease-risk strata, but TriatoScores suggest that the official classification likely underestimates risk in 42 municipalities. Of 152 municipalities failing to report triatomines in 2006–2019, two and 71 had TriatoScores corresponding to, respectively, high and moderate entomological risk. Conclusions TriatoScore can help control-surveillance managers to flexibly assess and stratify the entomological risk of Chagas disease at operationally relevant scales. Integrating eco-epidemiological, demographic, socioeconomic, or operational data (on, e.g., local-scale dwelling-infestation or vector-infection frequencies, land-use change and urbanization, housing conditions, poverty, or the functioning of control-surveillance systems) is also straightforward. TriatoScore may thus become a useful addition to the triatomine bug control-surveillance toolbox. Graphical abstract

Background: Aedes albopictus is a highly invasive species and an important vector of dengue and chikungunya viruses. Indigenous to Southeast Asia, Ae. albopictus has successfully invaded every inhabited continent, except Antarctica, in the past 80 years. Vector surveillance and control at points of entry (PoE) is the most critical front line of defence against the introduction of Ae. albopictus to new areas. Identifying the pathways by which Ae. albopictus are introduced is the key to implementing effective vector surveillance to rapidly detect introductions and to eliminate them. Methods: A literature review was conducted to identify studies and data sources reporting the known and suspected dispersal pathways of human-mediated Ae. albopictus dispersal between 1940–2020. Studies and data sources reporting the first introduction of Ae. albopictus in a new country were selected for data extraction and analyses. Results: Between 1940–2020, Ae. albopictus was reported via various dispersal pathways into 86 new countries. Two main dispersal pathways were identified: (1) at global and continental spatial scales, maritime sea transport was the main dispersal pathway for Ae. albopictus into new countries in the middle to late 20th Century, with ships carrying used tyres of particular importance during the 1980s and 1990s, and (2) at continental and national spatial scales, the passive transportation of Ae. albopictus in ground vehicles and to a lesser extent the trade of used tyres and maritime sea transport appear to be the major drivers of Ae. albopictus dispersal into new countries, especially in Europe. Finally, the dispersal pathways for the introduction and spread of Ae. albopictus in numerous countries remains unknown, especially from the 1990s onwards. Conclusions: This review identified the main known and suspected dispersal pathways of human-mediated Ae. albopictus dispersal leading to the first introduction of Ae. albopictus into new countries and highlighted gaps in our understanding of Ae. albopictus dispersal pathways. Relevant advances in vector surveillance and genomic tracking techniques are presented and discussed in the context of improving vector surveillance. [ABSTRACT FROM AUTHOR]

Simple Summary: The mosquito is the deadliest animal on earth, transmitting diseases that cause nearly a million deaths and >700 million infections each year. Yet only relatively few mosquito species are "vectors" that transmit diseases. Unfortunately, identifying these vector species is a difficult and time-consuming manual process. A promising and scalable solution involves "citizen science", whereby the general public provides images of mosquito specimens and breeding habitats using smartphones. However, data from such previous efforts have lacked the necessary integration for a thorough and global understanding of mosquito presence. Here, we standardize and combine data from multiple international citizen science apps—Mosquito Alert, iNaturalist, and GLOBE Observer's Mosquito Habitat Mapper and Land Cover—to aid researchers, mosquito control personnel, and policymakers. We also launched coordinated media campaigns that generated unprecedented numbers and types of observations, including successfully capturing the first images of targeted invasive and vector species. Additionally, we used citizen science imagery to develop artificial intelligence software to automatically identify the species and anatomical regions of mosquitoes. Ultimately, we establish a new surveillance system to serve as a united front to combat the ongoing threat of mosquito-borne diseases worldwide. Mosquito-borne diseases continue to ravage humankind with >700 million infections and nearly one million deaths every year. Yet only a small percentage of the >3500 mosquito species transmit diseases, necessitating both extensive surveillance and precise identification. Unfortunately, such efforts are costly, time-consuming, and require entomological expertise. As envisioned by the Global Mosquito Alert Consortium, citizen science can provide a scalable solution. However, disparate data standards across existing platforms have thus far precluded truly global integration. Here, utilizing Open Geospatial Consortium standards, we harmonized four data streams from three established mobile apps—Mosquito Alert, iNaturalist, and GLOBE Observer's Mosquito Habitat Mapper and Land Cover—to facilitate interoperability and utility for researchers, mosquito control personnel, and policymakers. We also launched coordinated media campaigns that generated unprecedented numbers and types of observations, including successfully capturing the first images of targeted invasive and vector species. Additionally, we leveraged pooled image data to develop a toolset of artificial intelligence algorithms for future deployment in taxonomic and anatomical identification. Ultimately, by harnessing the combined powers of citizen science and artificial intelligence, we establish a next-generation surveillance framework to serve as a united front to combat the ongoing threat of mosquito-borne diseases worldwide. [ABSTRACT FROM AUTHOR]

Simple Summary: The GLOBE Program's GLOBE Observer Mosquito Habitat Mapper is a free citizen science data collection tool that can be downloaded onto smartphones. The Mosquito Habitat Mapper encourages individuals to participate in locating and removing mosquito breeding habitats from use. An easy-to-use graphic interface enables users to report and describe mosquito breeding habitats, places with standing water where immature mosquitoes grow and develop. Citizen scientists are asked to determine if they see immature mosquitoes and if they wish to count and identify any mosquito larvae they see. In the last task, the user is asked to dump out or cover the standing water source, eliminating its use as a breeding habitat. In this way, the GLOBE Observer mobile app also supports the actions of individuals protecting their communities from mosquito-borne disease. In addition, all data reported by citizen scientists are publicly available. Scientists are accessing this data for a variety of research uses, including the development of automated techniques to recognize larvae and mosquito breeding sites from digital images. Since 2017, more than 32,000 Mosquito Habitat Mapper observations have been submitted by citizen scientists in 84 countries. The GLOBE Program's GLOBE Observer Mosquito Habitat Mapper is a no-cost citizen scientist data collection tool compatible with Android and iOS devices. Available in 14 languages and 126 countries, it supports mosquito vector surveillance, mitigation, and education by interested individuals and as part of participatory community surveillance programs. For low-resource communities where mosquito control services are inadequate, the Mosquito Habitat Mapper supports local health action, empowerment, and environmental justice. The tangible benefits to human health supported by the Mosquito Habitat Mapper have encouraged its wide adoption, with more than 32,000 observations submitted from 84 countries. The Mosquito Habitat Mapper surveillance and data collection tool is complemented by an open database, a map visualization interface, data processing and analysis tools, and a supporting education and outreach campaign. The mobile app tool and associated research and education assets can be rapidly deployed in the event of a pandemic or local disease outbreak, contributing to global readiness and resilience in the face of mosquito-borne disease. Here, we describe the app, the Mosquito Habitat Mapper information system, examples of Mosquito Habitat Mapper deployment in scientific research, and the outreach campaign that supports volunteer training and STEM education of students worldwide. [ABSTRACT FROM AUTHOR]

Simple Summary: Vector-borne diseases are often zoonotic and so a One Health approach must be employed in order to investigate and control them. Therefore, surveillance of arthropod vectors and pathogens among animal populations should complement human disease surveillance. Since traditional surveillance methods to collect arthropod vectors and conduct pathogen testing from animals can be challenging, data collection can be supplemented with citizen science approaches, where the general public is actively involved in collecting animals and/or samples. In this review, we discuss considerations for researchers to create a successful vector surveillance program using citizen science approaches with different stakeholders who own, have interests in, or work with animals. Many vector-borne diseases that affect humans are zoonotic, often involving some animal host amplifying the pathogen and infecting an arthropod vector, followed by pathogen spillover into the human population via the bite of the infected vector. As urbanization, globalization, travel, and trade continue to increase, so does the risk posed by vector-borne diseases and spillover events. With the introduction of new vectors and potential pathogens as well as range expansions of native vectors, it is vital to conduct vector and vector-borne disease surveillance. Traditional surveillance methods can be time-consuming and labor-intensive, especially when surveillance involves sampling from animals. In order to monitor for potential vector-borne disease threats, researchers have turned to the public to help with data collection. To address vector-borne disease and animal conservation needs, we conducted a literature review of studies from the United States and Canada utilizing citizen science efforts to collect arthropods of public health and veterinary interest from animals. We identified common stakeholder groups, the types of surveillance that are common with each group, and the literature gaps on understudied vectors and populations. From this review, we synthesized considerations for future research projects involving citizen scientist collection of arthropods that affect humans and animals. [ABSTRACT FROM AUTHOR]

Abstract Background Vector surveillance is essential in determining the geographical distribution of mosquito vectors and understanding the dynamics of malaria transmission. With the elimination of human malaria cases, knowlesi malaria cases in humans are increasing in Malaysia. This necessitates intensive vector studies using safer trapping methods which are both field efficient and able to attract the local vector populations. Thus, this study evaluated the potential of Mosquito Magnet as a collection tool for Anopheles mosquito vectors of simian malaria along with other known collection methods. Methods A randomized 4 × 4 Latin square designed experiment was conducted to compare the efficiency of the Mosquito Magnet against three other common trapping methods: human landing catch (HLC), CDC light trap and human baited trap (HBT). The experiment was conducted over six replicates where sampling within each replicate was carried out for 4 consecutive nights. An additional 4 nights of sampling was used to further evaluate the Mosquito Magnet against the “gold standard” HLC. The abundance of Anopheles sampled by different methods was compared and evaluated with focus on the Anopheles from the Leucosphyrus group, the vectors of knowlesi malaria. Results The Latin square designed experiment showed HLC caught the greatest number of Anopheles mosquitoes (n = 321) compared to the HBT (n = 87), Mosquito Magnet (n = 58) and CDC light trap (n = 13). The GLMM analysis showed that the HLC method caught significantly more Anopheles mosquitoes compared to Mosquito Magnet (P = 0.049). However, there was no significant difference in mean nightly catch of Anopheles mosquitoes between Mosquito Magnet and the other two trapping methods, HBT (P = 0.646) and CDC light traps (P = 0.197). The mean nightly catch for both An. introlatus (9.33 ± 4.341) and An. cracens (4.00 ± 2.273) caught using HLC was higher than that of Mosquito Magnet, though the differences were not statistically significant (P > 0.05). This is in contrast to the mean nightly catch of An. sinensis (15.75 ± 5.640) and An. maculatus (15.78 ± 3.479) where HLC showed significantly more mosquito catches compared to Mosquito Magnet (P

• Variations is explained by ecological and climatic characteristics than with the meteorological variations. • Only night temperatures would explain variations in oviposition activity. • Variables are strong affected by Ecoregional, and ecoregional effects are stronger in small towns than in large cities. • Ae. aegypti first recorded at High Monte ecoregion. Probably result of town size effect. (i. e. Cafayate a city has 18,000 inhabitants where Ae. aegypti was detected, and next town San Carlos (7000 habitants, 21 km away, 20 min by road). Mosquitoes were not detected. We strongly recommend carry out new studies to verify this. 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. [Display omitted] [ABSTRACT FROM AUTHOR]

There are increasing reports of the Asian malaria mosquito, Anopheles stephensi invading and spreading in Eastern Africa. We discuss the importance of these invasions in the context of broader challenges facing malaria control in Africa and argue against addressing it as an isolated problem. Anopheles stephensi is only one of multiple biological threats facing malaria control in the region—and is itself an indication of wide-ranging weaknesses in vector surveillance and control programs. Expanded investigations are needed in both urban and rural areas, especially in countries serviced by the Indian Ocean trade routes, to establish the full extent and future trajectories of the problem. More importantly, instead of tackling this vector species as a stand-alone threat, affected countries should adopt more integrated and multi-sectorial initiatives that can sustainably drive and keep out malaria. [ABSTRACT FROM AUTHOR]

We aimed to determine the presence and distribution of Borrelia burgdorferi sensu lato (s.l.) in Ixodes ricinus ticks collected from urbanized and wild areas in Kaylaka Park (Bulgaria). A total of 546 ticks were collected over three years (2017–2019). The presence of Borrelia in 334 of the collected I. ricinus was detected by dark-field microscopy (DFM) and two nested PCRs (nPCR) targeting the borrelial 5S-23S rRNA intergenic spacer and Flagellin B (FlaB) gene. DFM was performed on a total of 215 ticks, of which 86 (40%) were positive. PCR was performed on 153 of the ticks. In total, 42.5% of the 5S-23S rRNA intergenic spacer and 49% of FlaB were positive. Considering as positive any single tick in which Borrelia sp. was detected regardless of the used method, the infection rate reached 37% (10/27) in the nymphs and 48.5% (149/307) in the adults (48.7% (77/158) females, 48.3% (72/149) males). The incidence of B. burgdorferi infection in I. ricinus did not differ statistically significantly between female, male, and nymph. This study provides evidence that Lyme disease spirochetes are present in various regions of Kaylaka Park with extremely high prevalence in their vectors. [ABSTRACT FROM AUTHOR]

Background: Proper vector surveillance relies on the ability to identify species of interest accurately and efficiently, though this can be difficult in groups containing cryptic species. Culicoides Latreille is a genus of small biting flies responsible for the transmission of numerous pathogens to a multitude of vertebrates. Regarding pathogen transmission, the C. variipennis species complex is of particular interest in North America. Of the six species within this group, only C. sonorensis Wirth & Jones is a proven vector of bluetongue virus and epizootic hemorrhagic disease virus. Unfortunately, subtle morphological differences, cryptic species, and mitonuclear discordance make species identification in the C. variipennis complex challenging. Recently, single-nucleotide polymorphism (SNP) analysis enabled discrimination between the species of this group; however, this demanding approach is not practical for vector surveillance. Methods: The aim of the current study was to develop a reliable and affordable way of distinguishing between the species within the C. variipennis complex, especially C. sonorensis. Twenty-five putative microsatellite markers were identified using the C. sonorensis genome and tested for amplification within five species of the C. variipennis complex. Machine learning was then used to determine which markers best explain the genetic differentiation between species. This led to the development of a subset of four and seven markers, which were also tested for species differentiation. Results: A total of 21 microsatellite markers were successfully amplified in the species tested. Clustering analyses of all of these markers recovered the same species-level identification as the previous SNP data. Additionally, the subset of seven markers was equally capable of accurately distinguishing between the members of the C. variipennis complex as the 21 microsatellite markers. Finally, one microsatellite marker (C508) was found to be species-specific, only amplifying in the vector species C. sonorensis among the samples tested. Conclusions: These microsatellites provide an affordable way to distinguish between the sibling species of the C. variipennis complex and could lead to a better understanding of the species dynamics within this group. Additionally, after further testing, marker C508 may allow for the identification of C. sonorensis with a single-tube assay, potentially providing a powerful new tool for vector surveillance in North America. [ABSTRACT FROM AUTHOR]

Background. Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira that affects both humans and animals worldwide. Early detection of the pathogen in humans is crucial for early intervention and control of the progression of the disease to a severe state. It is also vitally important to be able to detect the presence of the pathogen in carrier animals to control the spread of the disease from the environment. Here we developed a simple and rapid loop-mediated isothermal amplification (LAMP) assay targeting the leptospiral secY gene. Results. Several reaction conditions of the LAMP reaction were optimized to ensure efficient amplification of the target DNA. The sensitivity of the developed LAMP assay obtained using a pure Leptospira culture was 2 - 104 copies of genomic DNA per reaction (equivalent to 0.1 ng) for a 40-minute reaction time. No cross-reactions were observed in the LAMP reaction against a series of non-leptospiral bacteria, indicating a specific reaction. The applicability of the LAMP assay was demonstrated on human blood and urine specimens collected from suspected leptospirosis patients and rat kidney specimens collected from suspected leptospirosis outbreak areas and high-risk areas. The developed LAMP assay demonstrated a higher detection rate for leptospiral DNA compared with the polymerase chain reaction (PCR) assay, possibly due to the presence of inhibitory substances, especially in rat kidney specimens, to which the PCR method is more susceptible. The present findings also highlight the importance of urine sample collection from patients for routine monitoring of the disease. Conclusions. In short, the developed LAMP assay can serve as a feasible alternative tool for the diagnosis of leptospirosis and be used for epidemiological and environ-mental surveillance of the disease, considering its robustness, rapidity, sensitivity, and specificity, as demonstrated in this study. [ABSTRACT FROM AUTHOR]

Background Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira that affects both humans and animals worldwide. Early detection of the pathogen in humans is crucial for early intervention and control of the progression of the disease to a severe state. It is also vitally important to be able to detect the presence of the pathogen in carrier animals to control the spread of the disease from the environment. Here we developed a simple and rapid loop-mediated isothermal amplification (LAMP) assay targeting the leptospiral secY gene. Results Several reaction conditions of the LAMP reaction were optimized to ensure efficient amplification of the target DNA. The sensitivity of the developed LAMP assay obtained using a pure Leptospira culture was 2 × 104 copies of genomic DNA per reaction (equivalent to 0.1 ng) for a 40-minute reaction time. No cross-reactions were observed in the LAMP reaction against a series of non-leptospiral bacteria, indicating a specific reaction. The applicability of the LAMP assay was demonstrated on human blood and urine specimens collected from suspected leptospirosis patients and rat kidney specimens collected from suspected leptospirosis outbreak areas and high-risk areas. The developed LAMP assay demonstrated a higher detection rate for leptospiral DNA compared with the polymerase chain reaction (PCR) assay, possibly due to the presence of inhibitory substances, especially in rat kidney specimens, to which the PCR method is more susceptible. The present findings also highlight the importance of urine sample collection from patients for routine monitoring of the disease. Conclusions In short, the developed LAMP assay can serve as a feasible alternative tool for the diagnosis of leptospirosis and be used for epidemiological and environmental surveillance of the disease, considering its robustness, rapidity, sensitivity, and specificity, as demonstrated in this study.