Risk of Arboviral Transmission and Insecticide Resistance Status of Aedes Mosquitoes during a Yellow Fever Outbreak in Ghana

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Yellow fever is endemic in Ghana. However, there is currently no structured vector control programme for the yellow vector, Aedes mosquitoes in Ghana. Knowledge of Aedes bionomics and insecticide susceptibility status is important to control the vectors. This study therefore sought todetermine Aedes vector bionomics and their insecticide resistance status during a yellow fever outbreak. Methods The study was performed in two yellow fever outbreak sites (Wenchi, Larabanga) and two non-outbreak sites (Kpalsogu, Pagaza) in Ghana. Immature Aedes mosquitoes were sampled from water-holding containers in and around human habitations. The risk of disease transmission was determined in each site using stegomyia indices. Adult Aedes mosquitoes were sampled using Biogents Sentinel (BG) traps, Human Landing Catch (HLC), and Prokopack (PPK) aspirators. Phenotypic resistance was determined with WHO susceptibility tests using Aedes mosquitoes collected as larvae and reared into adults. Knockdown resistance (kdr) mutations were detected using allele-specific multiplex PCR. Results Of the 2,664 immature Aedes sampled, more than 60% were found in car tyres. Larabanga, an outbreak site, was classified as a high-risk zone for the Yellow Fever outbreak (BI: 84%, CI: 26.4%). Out of 1,507 adult Aedes mosquitoes collected, Aedes aegypti was the predominant vector species (92%). A significantly high abundance of Aedes mosquitoes was observed during the dry season (61.2%) and outdoors (60.6%) ( P < 0.001). Moderate to high resistance to deltamethrin was observed in all sites (33.75% to 70%). Moderate resistance to pirimiphos-methyl (65%) was observed in Kpalsogu. Aedes mosquitoes from Larabanga were susceptible (98%) to permethrin. The F1534C kdr, V1016I kdr and V410 kdr alleles were present in all the sites with frequencies between (0.05-0.92). The outbreak sites had significantly higher allele frequencies of F1534C and V1016I respectively compared to non-outbreak sites ( P < 0.001). Conclusion This study indicates that Aedes mosquitoes in Ghana pose a significant risk to public health, and there is a need for continuous surveillance to inform effective vector control strategies. Aedes aegypti yellow fever Stegomyia indices Ghana Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Aedes mosquitoes represent an ever-growing threat to public health worldwide due to their ability to transmit many infectious arboviral pathogens such as Dengue, Chikungunya, Zika and Yellow Fever ( 1 ). Yellow fever (YF), an acute viral disease affecting humans and non-human primates (NHP), is caused by the yellow fever virus (YFV) ( 2 ). The virus is transmitted by the bite of infected female Aedes mosquitoes ( 3 ). Aedes aegypti is one of the vectors for yellow fever in Africa alongside Ae. albopictus which is known to be more invasive and also a competent vector for Dengue fever and Chikungunya ( 4 ). The World Health Organisation (WHO) has reported that forty-seven (47) countries in Africa are either endemic or have regions that are endemic for YF and other arboviral infections ( 5 ). Furthermore, the WHO advises countries that have Aedes mosquitoes but no evidence of viral transmission to identify local regions with high mosquito densities and make proper preparations to deal with any possible arboviral outbreaks ( 6 ). The disease affects over 200,000 people and causes about 30,000 deaths annually ( 7 ). In Africa, annually an estimated 21 million people are at risk ( 8 ). Yellow Fever is endemic in Ghana; this situation is exacerbated by the limited vaccine coverage, vector abundance and increasing insecticide resistance ( 9 – 12 ), creating a risk for onward transmission and amplification of the virus among unvaccinated populations ( 10 ). Major arboviral disease outbreaks have been recorded in Ghana since 1969 ( 10 ). A recent outbreak of yellow fever was experienced in Ghana in 2021 within the Savannah, Oti, Bono and Upper West regions. Reports from the Ghana Health Service, as of 4th December 2021, 202 suspected cases had been reported with 85 confirmed cases and 46 mortalities ( 13 ). Evidence of the presence and exposure to other arboviral diseases such as dengue fever and chikungunya have also been reported in Ghana ( 14 – 16 ). These reports show that arboviral pathogens are in circulation in Ghana and require the establishment of effective surveillance and vector control management strategies. Despite the increasing outbreaks of arboviral diseases and the high densities of the arboviral vectors in Africa, its control is given limited attention ( 8 ). The control of arboviral diseases majorly relies on vector control using insecticides coupled with larval source management and case management. Increasing insecticide resistance in Aedes mosquitoes poses a major challenge for vector control strategies. Resistance of the Aedes mosquitoes to insecticides has been reported in some West African countries like Senegal, Burkina Faso and Ghana ( 9 , 11 , 17 , 18 ). Aedes mosquitoes populations in Ghana have been found to be resistant to several public health insecticides including pyrethroids, organochlorines and carbamates ( 9 , 11 , 12 , 19 ). Target-site mutations such as V410L, V1016I and F1534C have been found in pyrethroid-resistant Aedes mosquitoes from Ghana and other countries ( 9 , 20 – 23 ). Vector and insecticide susceptibility surveillance is still crucial in reducing the global burden of arboviral infections ( 24 ). However, there is a paucity of data on the risk of transmission and insecticide susceptibility status of these arboviral disease vectors in Ghana. The current study sought to determine the risk of arboviral transmission and insecticide susceptibility status of the Aedes mosquitoes in the selected yellow fever outbreak and non-outbreak areas in Ghana. Methods Study Sites This study was conducted in four ( 4 ) sites, two yellow fever outbreak areas [Larabanga (9°5′0″N, 1°49′0″W) and Wenchi (7°33′33″N 1°55′45″W)] and two non-outbreak areas [Kpalsogu (9°33′45.2″N, 1°01′54.6″W) and Pagaza (9°22′33.34″N, 0°42′29.67″W)] as control areas. Of the four sites, three ( 3 ) were rural areas (Larabanga, Kpalsogu, Pagaza) and one ( 1 ) urban area (Wenchi). Sampling of mosquitoes was done during the dry (April – June) and rainy (August – October) seasons. The control sites were selected due to their proximity to the districts that had the outbreak; however, no YF cases had been recorded in these villages during the yellow fever outbreak. Figure 1: Map of Ghana indicating the study areas. Characterization of Aedes breeding habitats and abundance of Aedes larvae Larval sampling was performed in each study site, to characterise the breeding habitats and abundance of immature Aedes mosquitoes. During the larval surveys, the habitat type, its location in a household (indoor or outdoor), and its physical characteristics were recorded. Eight container types were classified based on their use and material: car tyres, discarded containers, drinking pots, drums, tanks, and buckets. Discarded containers and drinking pots were 50–100 L capacity containers which included broken jars, bottles, small plastic food containers, tins, plates, cans, cooking pots, drinking troughs and broken pots made of clay, plastic or metal. Drums were defined as 100-500L capacity plastic water storage containers. Tanks were 100–500 L capacity water storage containers made of metal or concrete. Buckets included 10–25 L water storage containers made of metal or plastic. In almost all the sites, access to pipe-borne water was a big challenge, therefore, households tend to store water in storage containers, pots, drums etc for long-term use. These containers may favour the breeding of the Aedes vector. Coordinates of all collection points were recorded using a GPSMAP® 60CSx geographical position system (GPS) instrument (Garmin International, Inc., Olathe, Kansas, USA). All larval samples were transported to the insectary at the Department of Medical Microbiology, University of Ghana, where they were raised to adults under suitable conditions (temperature: 27 ± 2°C, 75 ± 10% relative humidity). The larvae were fed on TetraMin Baby fish food (Tetra Werke, Melle, Germany). Emerged adults were morphologically identified using standard taxonomic keys ( 25 ). Figure 2: Larval habitats encountered during larval sampling. a Drinking pot, b Tank, c Car tyre, g Discarded container, e Drum f Bucket Determination of Stegomyia Indices The extent of infestation by Aedes mosquitoes was estimated using the classical Stegomyia indices including BI, Breteau Index, (the number of positive containers per 100 surveyed houses); HI; House Index, (percentage of houses positive for Aedes larvae or pupae); CI; Container Index, (the percentage of containers positive per 100 houses inspected). These larval and pupal indices continue to be the predominant and frequent measures used to evaluate vector prevalence because catching adult mosquitoes is time-consuming and requires access to private land ( 12 ). Stegomyia indices are quantitative indicators of the risk of transmission. Using the WHO criteria, the risk of YF at each site was assessed as follows: In an area where BI, HI, and CI exceeded 50%, 35% and 20% respectively, the risk of Ae aegypti -transmitted Viral Haemorrhagic Fever (VHF) was considered to be high; an area where BI was between 5% and 50%, the density of Ae. aegypti was considered to be sufficient to promote an outbreak of VHF disease; an area where BI, HI and CI were less than 5%, 4% and 3% respectively, it was considered to be unlikely for YF transmission to occur ( 26 ). Adult Aedes mosquito sampling The spatio-temporal distribution of adult Aedes mosquitoes was determined by sampling indoors and outdoors using three methods; Biogent sentinel 2 traps (BG traps), Human landing catches (HLC), and prokopack aspiration (PPK) (John W. Hock Company, Gainesville, U.S.A.) ( 27 ). Geographical coordinates of each house sampled were taken. Cross-sectional surveys were undertaken in the dry season (March to May 2022) and in the rainy season (August to October 2022). Sixteen houses were randomly selected for each sampling method at each site (4 houses per day). Biogent sentinel traps were set both indoors (living rooms and bedrooms) and outdoors (open, verandas, or under a shed/tree where people sit to chat) about 5 m from the house) during the times 3:00–7:00 pm. The BG traps were baited with carbon dioxide (CO 2 ) which was produced from a mixture of 17.5 g yeast (Angel Yeast (Egypt) Co. Ltd.), 250 g sugar in 1 litre of water (van Loon et al. , 2015). After the 4 hours, mosquitoes trapped were carefully removed, placed in a cooler box containing ice and then transported to the insectary. The HLC method was also used to sample host-seeking adult Aedes mosquitoes. On each day, one trained volunteer was positioned indoors and another outdoors and collected mosquitoes from 3:00–6:00 pm. Collected Aedes were placed in well-labelled paper cups, and transported to the insectary for identification and further processing. Prokopack aspirations were used to sample resting mosquitoes. Sampling for Aedes mosquitoes was done indoors and outdoors. Sampled adult Aedes mosquitoes were knocked down with chloroform and preserved in Eppendorf tubes containing silica gel. Morphological identification of A edes mosquitoes All adult Aedes mosquitoes from the adult sampling, and those used for the susceptibility testing (Aedes larvae collected in the study sites and raised to adults), were identified morphologically using the taxonomic keys by Huang ( 25 ). Insecticide susceptibility tests Insecticide susceptibility test was conducted using WHO tubes to determine phenotypic resistance according to WHO criteria ( 28 ). Adult female Aedes mosquitoes that were 3–5-day-old were exposed to papers impregnated with permethrin (0.75%), deltamethrin (0.05%), and pirimiphos-methyl (0.25%). Larvae and pupae collected from the larval sampling as well as the Ovitraps that were set were used for the WHO susceptibility testing. Though these doses are not the recommended doses for evaluating the susceptibility of Aedes mosquitoes, they are the most commonly used ( 11 , 29 ). The knockdown time was recorded every 10 min during the 60-minute exposure period. Mortalities were recorded after a 24-hour recovery period. Alive (resistant) and dead (susceptible) mosquitoes were stored in absolute ethanol for later DNA analysis. Genotyping of kdr mutations in Aedes aegypti populations A sub-sample of 242 phenotypically pyrethroid resistant and susceptible Aedes mosquitoes were genotyped of kdr mutations, F1534C, V1016I and V410L. Total DNA was extracted from whole mosquitoes using the DNeasy Tissue Kit (Qiagen, In USA). Genotyping of the kdr mutations was done using allele-specific multiplex PCR according to well-described protocols of Villanueva-Segura et al. ( 30 ) Results Distribution and abundance of Aedes larval habitats. A total of 535 larval habitats with 86 positive breeding habitats were surveyed from all the study sites, comprising six ( 6 ) different habitat types over the entire sampling period. Overall, the most abundant habitat type was car tyres [59.3%, n = 51/86], whereas the least were tanks [2.3%, n = 2/86] and buckets [2.3%, n = 2/86] (Table 1 ). A significantly higher abundance of larval habitats was encountered during the rainy season [62.8%, n = 54/86] than in the dry season [37.2%, n = 32/86] ( X 2 = 18.5035, df = 5, P = 0.002). Within the 4 sites sampled, a significantly higher proportion (99.8%, 85/86) of larval habitats were encountered outdoors as compared to indoors (1.2%, 1/86) ( X 2 = 42.4941, df = 5, P < 0.001). Table 1 Larval habitat distribution abundance per season and location. Dry Season Rainy Season Habitat Type Indoor (%) Outdoor (%) Indoor (%) Outdoor (%) Total (%) Bucket 0 0 1 (100) 1 (1.9) 2 (2.3) Discarded container 0 1 (3.1) 0 10 (18.9) 11 (12.8) Drinking pot 0 1 (3.1) 0 14 (26.4) 15 (17.5) Drum 0 1 (3.1) 0 4 (7.5) 5 (5.8) Tank 0 1 (3.1) 0 1 (1.9) 2 (2.3) Car Tyre 0 28 (87.6) 0 23 (43.4) 51 (59.3) Total 0 (0.00) 32 (37.21) 1 (1.16) 53 (61.63) 86 (100) Larval densities of Aedes immature A total of 2,664 Aedes immatures were collected over the entire sampling period, of which the most productive habitat type was car tyres 65.1% (1,734/2664), whereas the least productive was tanks 1.1% (30/2664) (Table 2 ). Aedes larval abundance was slightly higher at 1,342 (50.4%) in the rainy season as compared to the dry season at 1,322 (49.6%) ( X 2 = 37.1991, df = 28, P = 0.115). During both seasons, the highest larval abundance was observed in Larabanga, a yellow fever outbreak site [n = 1,472 (55.3%)], whereas the least was observed in Wenchi, also an outbreak site [n = 217 (8. 1%)], Table 2 . A significantly higher abundance of Aedes immatures were collected outdoors [n = 2,626 (98.6%)] as compared to [n = 38 (1.4%)] those collected indoors ( X 2 = 86.000, df = 28, P < 0.001), (Table 2 ). Table 2 The seasonal distribution of Aedes immatures across the study sites. Wenchi Larabanga Kpalsogu Pagaza Location Dry Rainy Dry Rainy Dry Rainy Dry Rainy Indoor 0 0 0 0 0 38 0 0 Outdoor 145 72 832 640 0 392 345 200 Container Type Bucket 0 0 0 0 0 38 0 42 Discarded container 20 0 0 75 0 122 0 101 Drinking pot 0 0 0 75 0 210 30 57 Drum 10 0 0 75 0 45 0 0 Tank 0 0 15 15 0 0 0 0 Car Tyre 115 72 817 400 0 15 315 0 Total 145 72 832 640 0 430 345 200 Larval Indices (Stegomyia Indices) Larval Indices calculated using the WHO formula for each study site were used to make inferences about the risk of transmission of yellow fever in the outbreak and non-outbreak zones studied. From the results obtained, in Larabanga the indices (BI: 84% CI:26.4%) exceeded the threshold and can be classified as a high-risk zone for yellow fever transmission as compared to all the other study areas, Wenchi (BI:42.3% CI:19.3% HI:23.1%), Kpalsogu (BI:37.7% CI:12.1% HI:6.7%) and Pagaza (BI: 30.2% CI:8.9% HI:9.4%) that had values within the range sufficient to promote an outbreak. These are shown in Fig. 3. Figure 3: Stegomyia Indices per study site Spatio-temporal distribution of adult Aedes mosquitoes Overall, a total of 1,507 adult Aedes mosquitoes were collected from all the study areas, with Aedes aegypt i [92%, n = 1386/1507] as the predominant species, followed by Aedes formosus [8%, n = 121/1507]. A high abundance of Aedes mosquitoes was collected in Larabanga, a yellow fever outbreak site [ Aedes aegypti 884/1386 (63.8%); Aedes formosus 112/121 (92.3%)], whereas the least abundance was collected in Pagaza, a non-yellow fever outbreak site [ Aedes aegypti 75/1386 (5.4%); Aedes formosus 9/121 (7.7%)] ( X 2 = 52.061, df = 3, P < 0.001). A high abundance of Aedes mosquitoes was sampled outdoors [n = 914 (60.7%)] as compared to indoor collections [n = 593 (39.3%); X 2 = 68.38, df = 1, P < 0.01)]. A high abundance of adult Aedes mosquitoes was sampled for both indoor [n = 342 (34.3%)] and outdoor [n = 654 (65.7%)] collections in Larabanga whereas, the least recorded abundance was in Pagaza [Indoor (n = 45 (53.6%); Outdoor (n = 39, 46.4%; X 2 = 32.021, df = 3, P < 0.001), Table 3 . Table 3 Adult Aedes abundance per study site and location Study Sites Aedes aegypti Aedes formosus Indoor Outdoor Total per Site No. (%) No. (%) No. (%) No. (%) No. (%) Wenchi 325 (100) 0 (0.0) 159 (48.9) 166 (51.1) 325 (21.5) Larabanga 884 (88.0) 112 (12.0) 342 (34.3) 654 (65.7) 996 (66.6) Kpalsogu 102 (100) 0 (0.0) 47 (46.1) 55 (53.9) 102 (6.7) Pagaza 75 (89.0.) 9 (11.0) 45 (53.6) 39 (46.4) 84 (5.6) Total 1386 (92.0) 121 (8.0) 593 (39.3) 914 (60.7 1507 (100) Adult Aedes mosquitoes were more predominant in the dry season [n = 922/1507 (61.2%)] than in the rainy season [n = 585/1507 (38.8%)], ( X 2 = 75.36, df = 1, P < 0.001). In the dry season, the highest abundance of adult Aedes mosquitoes was recorded in Larabanga [n = 289/585, (49.4%)], followed by those from Wenchi [n = 189/585, (32.3%)], both being yellow fever outbreak sites. Mosquitoes from the non-yellow fever outbreak sites were the least, Pagaza [n = 70/585, (12.0%)] and Kpalsogu [n = 37/585, (6.3%)]. Similarly, during the rainy season, the highest abundance of Aedes mosquitoes was recorded in Larabanga [n = 707/922, (76.7%)], followed by Wenchi [n = 136/922 (14.8%)], the yellow fever outbreak sites. There were small numbers of Aedes mosquitoes sampled in the non-yellow fever outbreak sites of Kpalsogu [n = 65/922, (7.0%)] and Pagaza [n = 14/922, (1.5%)]. These are shown in Fig. 4. Figure 4: Seasonal abundance of adult Aedes mosquitoes Seasonal abundance of adult Aedes mosquitoes per trap type Three separate traps namely, HLC, PPK, and BG were used to collect a total of 1507 adult Aedes mosquitoes from all the study sites. A total of 601 (39.9%) Aedes mosquitoes were collected using HLC [dry = 280 (46.6%); rainy = 321 (53.4%)], PPK 443 (29.4%) [dry = 183 (41.3%); rainy = 260 (58.7%)] and BG 463 (30.7%) [dry = 122 (26.3%); rainy = 341 (73.7%)]. Of the 796 female adult mosquitoes caught with PPK and BG, 575 [PPK = 295 (66.6%); BG = 280 (60.5%)] were unfed, 177 [PPK = 77 (17.4%) BG = 100 (21.6%)] were blood-fed, half-gravid [PPK = 12 (2.7%); BG = 0] and gravid [PPK = 18 (4.1%); BG = 14 (3.0%)], ( X 2 = 46.745, df = 2, P < 0.001) (Fig. 5). Figure 5: Season abundance of Aedes mosquitoes per trap type (In-Indoor; OUT-Outdoor; BG-Biogents-2 sentinel traps; HLC-Human Landing Catches; PPK-Prokopack aspiration) Insecticide susceptibility status of Aedes mosquitoes Bioassay results showed resistance to deltamethrin at x1 concentration across all the study sites (33.75% − 70%) (Fig. 6). However, susceptibility to deltamethrin at x5 was observed across all sites. The mosquitoes showed possible resistance to permethrin at x1 in Kpalsogu (95%), Pagaza (96.5%), and susceptibility in Larabanga (98%). Resistance to pirimiphos-methyl at x1 was shown in Aedes mosquitoes in Kpalsogu (63.75%) and possible resistance was observed in Pagaza (93.75%) and Larabanga (92.5%). For Wenchi, for logistical challenges, not many larvae were sampled for the determination of insecticide susceptibility, so only deltamethrin was run. These results are shown in Fig. 6. Figure 6: Mortalities of Aedes mosquitoes exposed to different insecticides in the study sites. Genotypic mutations associated with resistance in Aedes aegypti A subset of 242 Ae. aegypti obtained from the phenotypic assays were genotyped for the F1534C, V1016I and V410L kdr mutations. About 20–30 mosquito samples were selected from each site per insecticide paper per concentration for the genotypic resistance determination. The F1534C mutation was detected in moderate to high allele frequencies in Ae. aegypti mosquitoes exposed to pyrethroid insecticides. Aedes aegypti from Wenchi that were exposed to deltamethrin, had a significantly high allele frequency of F1534C mutation (0.92) compared to mosquitoes from the non-outbreak sites, Pagaza (0.19) and Kpalsogu (0.35) ( χ2 = 50.50, df = 3, P < 0.001). For the V1016I mutation, low to moderate allele frequencies (0.23 to 0.54) were observed in Ae. aegypti mosquitoes exposed to both deltamethrin and permethrin except for Larabanga, an outbreak site, where mosquitoes had a high allele frequency of 0.77. However, there was a significant difference in the frequency of V1016I mutation in outbreak and non-outbreak sites ( P < 0.05). For the V410L mutation, there was no significant differences in the frequency of the mutations between outbreak sites and non-outbreak sites with low allele frequencies ranging from 0.05 to 0.15. The genotypes and allele frequencies of each kdr mutation are shown in Table 4 . Table 4 Number of genotypes and frequencies of the F1534C, V1016I and V410L mutation in the voltage-gated sodium channel gene of Aedes aegypti mosquitoes. F1534C V1016I V410L Insecticide Site Description Study site n CC FF FC Allele Freq. II VV VI Allele Freq. LL VV VL Allele Freq. Deltamethrin Non-outbreak Pagaza 60 6 43 11 0.19 3 22 35 0.34 0 45 15 0.13 Kpalsogu 44 12 25 7 0.35 1 1 42 0.5 3 40 1 0.08 Outbreak Larabanga 30 8 13 9 0.42 2 0 28 0.53 0 27 3 0.05 Wenchi 52 45 1 6 0.92 8 4 40 0.54 3 46 3 0.09 Total 186 71 82 33 14 27 145 6 160 22 Permethrin Non-outbreak Pagaza 32 8 21 3 0.3 0 17 15 0.23 2 29 1 0.15 Kpalsogu 0 0 0 0 0 0 0 0 0 0 0 0 0 Outbreak Larabanga 24 5 16 3 0.27 13 0 11 0.77 0 24 0 0 Wenchi 0 0 0 0 0 0 0 0 0 0 0 0 0 Total 56 13 37 6 13 17 26 2 53 1 Abbreviations: VV, wild type (susceptible); VL, heterozygotes; LL, mutant (resistant); VI, heterozygotes; II, mutant (resistant); FF, wild type (susceptible); FC, heterozygotes; CC, mutant (resistant); n sample size. Discussion Due to their capacity to spread a variety of arboviral infections like dengue, chikungunya, Zika, and yellow fever, Aedes mosquitoes pose an increasing hazard to public health on a global scale (31). Ghana is endemic for yellow fever with the most recent outbreak occurring in 2021 (13). Hence, the need to monitor the densities and insecticide susceptibility status of Aedes mosquitoes in the country. This study sought to determine the risk of arboviral transmission and insecticide resistance status of Aedes mosquitoes in a yellow fever outbreak and non-outbreak areas in Ghana. From the data obtained, car tyres were the most productive habitats for Aedes mosquitoes seasonally. Moreover, the outbreak area (Larabanga) had a high Stegomyia indices value and hence satisfied the WHO criteria for a high-risk zone for yellow fever transmission. Adult Aedes aegypti was the predominant vector sampled in all the study sites, with high abundance in the dry season and from outdoor collection. The distribution of larval habitat types varied significantly between seasons. Findings from this study showed that car tyres were responsible for over 60% of Aedes immatures over the entire sampling period. Car tyres seems to provide an optimal temperature, humidity and sufficient light intensity to ensure larvae development (32). This finding is consistent with studies by Owusu-Asenso et al. (11) in Ghana and Kamgang et al. (33) in the Central African Republic who also found car tyres as the most conducive and productive breeding habitat for Aedes aegypti . In the current study, results from the larval indices indicate that Larabanga had values that exceeded the WHO threshold and hence is a high-risk zone for arboviral pathogen transmission. This is similar to a previous study by Appawu et al . (10) in Larabanga, where the Stegomyia indices exceeded the threshold and was considered as a high-risk zone for yellow fever transmission. Furthermore, Stegomyia indices values within the WHO criteria reported in Wenchi, Kpalsogu and Pagaza were sufficient to promote an outbreak within these sites. These findings imply that inhabitants within these study sites are at risk of yellow fever infection, Hence, there is a need to employ preventive measures through vaccination and effective vector control strategies. Seasonal variation in population density is common seasonally for Aedes mosquitoes due to their sensitivity to changes in temperature and rainfall (34). This study found a significantly higher abundance of Aedes immature during the rainy season. The development of mosquitoes, their survival and the effective transmission of pathogens are influenced by humidity, temperature and rainfall (35,36). Sufficient humidity and rainfall influence the breeding sites, increasing vector populations (37). Hence, an increase in breeding sites may explain the observed increase in Aedes immatures in the rainy season. This finding corroborates with studies in Ghana by Owusu-Asenso et al. (11) and in Kenya by Ndenga et al. (38). Their findings showed high densities of Aedes immatures in the rainy season. However, this finding was contrary to another study in Ghana where Aedes larvae were found predominantly in the dry season. He reported that during drought conditions, the surge in the storage of water creates more breeding habitats for Aedes mosquitoes, causing an increase in their abundance (10). It was observed that adult Aedes mosquitoes were predominantly collected during the dry season and in outdoor settings. Warmer temperatures enhance the growth of mosquitoes and viral replication (34). The life-limiting elements of latitude, altitude, temperature, rainfall, humidity, season, habitat, and dispersal have an impact on the distribution and population of Aedes mosquitoes (10). Finding more adult Aedes mosquitoes outside may suggest their exophilic nature as reported in previous studies in Ghana and Kenya (11,39). It is important to note that people spending more time outdoors compared to indoors influences the biting and feeding behaviour of Aedes mosquitoes (38). Studies in Ghana have also suggested that Aedes mosquitoes often rest outdoors before and after blood feeding (40) and were more abundant from outdoor collection in the dry season (10). It was observed in this study that the majority of the adult Aedes mosquitoes were Aedes aegypti which is responsible for yellow fever transmission in Ghana and can transmit other arboviral pathogens such as dengue fever virus (12). There have been previous reports of dengue viral infections in children in Ghana (14) and exposure to dengue and chikungunya (15,16,41) that show the role of Aedes aegypti in the transmission of multiple arboviral pathogens in Ghana, which cannot be overlooked. In this study, Aedes mosquitoes across the sites showed resistance to Deltamethrin. This might be due to the indirect impact of the use of insecticides for public health vector control such as the use of Long-Lasting Insecticidal Nets (LLINs) and IRS, as well as pesticide use in agriculture (11,42,43). This finding is similar to that reported in a study conducted in Ghana (Abdulai et al., 2023). Whereas Aedes mosquitoes collected from Pagaza and Kpalsogu showed suspected resistance to Permethrin, samples from Larabanga were found to be susceptible to Permethrin. Similarly, pyrethroid resistance has also been reported in Aedes aegypti populations from Ghana and other West African countries ((11,17,18,29) Aedes mosquitoes in this study were also resistant or possibly resistant to Pirimiphos-methyl. The findings suggest that resistance to this organophosphate by the Aedes population has increased. In this study, the F1534C and V1016I kdr mutations were high in frequencies in resistant and susceptible Aedes mosquitoes while V410L kdr mutation showed low frequencies. Although suspected resistance and susceptibility to permethrin were recorded in the Aedes mosquitoes from the same population, there is the risk of resistance developing over time due to the high frequencies of the F1534C and V1016I kdr mutations. Pyrethroid resistance in Aedes aegypti is a worldwide challenge for mosquito control due to its use for insecticide-treated nets and indoor residual spraying (44). Similarly, other studies in Ghana have found high frequencies of V1016I and F1534C kdr mutations in both pyrethroid-susceptible and resistant Aedes mosquitoes collected in Ghana (9,21). This suggests that other resistance mechanism such as metabolic resistance may be involved in insecticide resistance of Aedes aegypti populations in Ghana. Hence, further studies are needed to elucidate the mechanisms mediating insecticide resistance in Aedes mosquitoes in Ghana. Conclusion Our findings reveal that Aedes immatures were most abundant in the dry season and car tyres were the most productive habitat type. Aedes aegypti were the predominant species found. The results showed that Larabanga, the epicentre of the yellow fever outbreak is a high-risk zone for arboviral pathogen transmission. In addition, high phenotypic and genotypic resistance was observed in Aedes mosquito populations in Ghana. Surveillance of Aedes mosquito bionomics and insecticide susceptibility in Ghana is crucial to help in the development of arboviral vector control strategies to control and prevent arboviral outbreaks in Ghana. Abbreviations Ae .: Aedes ; DNA: Deoxyribonucleic acid; PCR: polymerase chain reaction; PMI: President Malaria Initiative; WHO: World Health Organization; YF: Yellow Fever; HLC: Human Landing Catch; PPK: Procopack; BG: Biogent sentinel trap; GHS: Ghana Health Service; HI: House Index; CI: Container Index; BI: Breateau Index; VHF: Viral Haemorrhagic Fever; GPS: Geographical position system Declarations Acknowledgement Weappreciate the management and staff of PMI-Vector Link Tamale, especially Mr Osei Akuoko, Mr Imoro Nurudeen, Mr Cosmos Pambit Zong and Mr Abdullai Abdul-Rahman for granting permission to use their facility and resources to carry out this project. Special thanks to the Centre for Vector-borne Disease Research (CVBDR) team at the Department of Medical Microbiology, University of Ghana Medical School, for the support and technical advice during this project. We also appreciate all the indigenes in the various study sites who offered field assistance. Funding This study was supported by grants from the National Institute of Health (D43 TW 011513). Availability of data and materials All the data supporting this study are included in the article. Ethical Consideration Ethical approval for this study was obtained from the University of Ghana, College of Health Sciences (CHS) Ethics and Protocol Review Committee (EPRC) (CHS-Et/M.11- P4.1/2021-2022). Meetings were held in each study site with chiefs and the residents to introduce the research study to the public. All study participants recruited for mosquito collection in this study were adults (18 years old and above). A written and signed consent was obtained from all adults who volunteered to participate in HLC. They were then trained before sampling of mosquitoes. A copy of the signed consent form was given to each of the HLC volunteers and another was kept in a locked cabinet with restricted access in offices at The Department of Medical Microbiology, University of Ghana Medical School. Verbal consent was obtained from household heads to sample mosquitoes in their houses and compounds. Authors’ contribution MOA, SKA and YAA were responsible for the study design, supervised the data collection, and contributed to the writing of the manuscript. MOA, IKS, AA, CMO-A, ED and ARM carried out the data collection, laboratory work and data analysis. MOA, CMO-A and AA drafted and revised the manuscript. All authors read and approved the final copy of the manuscript. Conflict of interest The authors declare no conflict of interest. References WHO. Geneva, World Health Organisation. 2016 [cited 2022 Jul 20]. Monitoring and managing insecticide resistance in Aedes mosquito populations, interim guidance for entomologists. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4271509","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":291813783,"identity":"909e708d-18eb-4e46-a4d9-22a0d18dfe11","order_by":0,"name":"Margaret Owusu-Akyaw","email":"","orcid":"","institution":"Department of Medical Microbiology, Centre for Vector-Borne Disease Research, University of Ghana","correspondingAuthor":false,"prefix":"","firstName":"Margaret","middleName":"","lastName":"Owusu-Akyaw","suffix":""},{"id":291813784,"identity":"2ce7c1c7-7fed-477c-af47-b7e1d7d7b883","order_by":1,"name":"Christopher Mfum 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site\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-4271509/v1/7ec6979b9902187aafb2802e.png"},{"id":54955811,"identity":"ddb575d7-1fcc-4144-bdb1-546c32598bff","added_by":"auto","created_at":"2024-04-19 07:12:25","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":26575,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSeasonal abundance of adult \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAedes\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e mosquitoes\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-4271509/v1/0e9d7905a5918b71793b66bd.png"},{"id":54955369,"identity":"1d232534-d643-48ba-84aa-64fe48d464fd","added_by":"auto","created_at":"2024-04-19 07:04:25","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":31370,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSeason abundance of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAedes\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e mosquitoes per trap type (In-Indoor; OUT-Outdoor; BG-Biogents-2 sentinel traps; HLC-Human Landing Catches; PPK-Prokopack aspiration)\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-4271509/v1/aa98a0bbe2ebf12f81332a33.png"},{"id":54955371,"identity":"435da139-b1d9-4ed1-8958-205ce0235ba3","added_by":"auto","created_at":"2024-04-19 07:04:25","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":24915,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMortalities of \u003c/strong\u003e\u003cem\u003e\u003cstrong\u003eAedes\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e mosquitoes exposed to different insecticides in the study sites.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Fig6.png","url":"https://assets-eu.researchsquare.com/files/rs-4271509/v1/f986f6d884638ad8b2793059.png"},{"id":54956277,"identity":"9eccc2cf-4ede-4fbe-990a-a197f068e973","added_by":"auto","created_at":"2024-04-19 07:20:26","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3588089,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4271509/v1/044881eb-1414-4e83-b952-615bcbf80ad7.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Risk of Arboviral Transmission and Insecticide Resistance Status of Aedes Mosquitoes during a Yellow Fever Outbreak in Ghana","fulltext":[{"header":"Introduction","content":"\u003cp\u003e \u003cem\u003eAedes\u003c/em\u003e mosquitoes represent an ever-growing threat to public health worldwide due to their ability to transmit many infectious arboviral pathogens such as Dengue, Chikungunya, Zika and Yellow Fever (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). Yellow fever (YF), an acute viral disease affecting humans and non-human primates (NHP), is caused by the yellow fever virus (YFV) (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). The virus is transmitted by the bite of infected female \u003cem\u003eAedes\u003c/em\u003e mosquitoes (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). \u003cem\u003eAedes aegypti\u003c/em\u003e is one of the vectors for yellow fever in Africa alongside \u003cem\u003eAe. albopictus\u003c/em\u003e which is known to be more invasive and also a competent vector for Dengue fever and Chikungunya (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e). The World Health Organisation (WHO) has reported that forty-seven (47) countries in Africa are either endemic or have regions that are endemic for YF and other arboviral infections (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e). Furthermore, the WHO advises countries that have \u003cem\u003eAedes\u003c/em\u003e mosquitoes but no evidence of viral transmission to identify local regions with high mosquito densities and make proper preparations to deal with any possible arboviral outbreaks (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e). The disease affects over 200,000 people and causes about 30,000 deaths annually (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e). In Africa, annually an estimated 21\u0026nbsp;million people are at risk (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eYellow Fever is endemic in Ghana; this situation is exacerbated by the limited vaccine coverage, vector abundance and increasing insecticide resistance (\u003cspan additionalcitationids=\"CR10 CR11\" citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e), creating a risk for onward transmission and amplification of the virus among unvaccinated populations (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e). Major arboviral disease outbreaks have been recorded in Ghana since 1969 (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e). A recent outbreak of yellow fever was experienced in Ghana in 2021 within the Savannah, Oti, Bono and Upper West regions. Reports from the Ghana Health Service, as of 4th December 2021, 202 suspected cases had been reported with 85 confirmed cases and 46 mortalities (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e). Evidence of the presence and exposure to other arboviral diseases such as dengue fever and chikungunya have also been reported in Ghana (\u003cspan additionalcitationids=\"CR15\" citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e). These reports show that arboviral pathogens are in circulation in Ghana and require the establishment of effective surveillance and vector control management strategies.\u003c/p\u003e \u003cp\u003eDespite the increasing outbreaks of arboviral diseases and the high densities of the arboviral vectors in Africa, its control is given limited attention (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). The control of arboviral diseases majorly relies on vector control using insecticides coupled with larval source management and case management. Increasing insecticide resistance in Aedes mosquitoes poses a major challenge for vector control strategies. Resistance of the \u003cem\u003eAedes\u003c/em\u003e mosquitoes to insecticides has been reported in some West African countries like Senegal, Burkina Faso and Ghana (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e). \u003cem\u003eAedes\u003c/em\u003e mosquitoes populations in Ghana have been found to be resistant to several public health insecticides including pyrethroids, organochlorines and carbamates (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e). Target-site mutations such as V410L, V1016I and F1534C have been found in pyrethroid-resistant Aedes mosquitoes from Ghana and other countries (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan additionalcitationids=\"CR21 CR22\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eVector and insecticide susceptibility surveillance is still crucial in reducing the global burden of arboviral infections (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e). However, there is a paucity of data on the risk of transmission and insecticide susceptibility status of these arboviral disease vectors in Ghana. The current study sought to determine the risk of arboviral transmission and insecticide susceptibility status of the \u003cem\u003eAedes\u003c/em\u003e mosquitoes in the selected yellow fever outbreak and non-outbreak areas in Ghana.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy Sites\u003c/h2\u003e \u003cp\u003eThis study was conducted in four (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e) sites, two yellow fever outbreak areas [Larabanga (9\u0026deg;5\u0026prime;0\u0026Prime;N, 1\u0026deg;49\u0026prime;0\u0026Prime;W) and Wenchi (7\u0026deg;33\u0026prime;33\u0026Prime;N 1\u0026deg;55\u0026prime;45\u0026Prime;W)] and two non-outbreak areas [Kpalsogu (9\u0026deg;33\u0026prime;45.2\u0026Prime;N, 1\u0026deg;01\u0026prime;54.6\u0026Prime;W) and Pagaza (9\u0026deg;22\u0026prime;33.34\u0026Prime;N, 0\u0026deg;42\u0026prime;29.67\u0026Prime;W)] as control areas. Of the four sites, three (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e) were rural areas (Larabanga, Kpalsogu, Pagaza) and one (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e) urban area (Wenchi). Sampling of mosquitoes was done during the dry (April \u0026ndash; June) and rainy (August \u0026ndash; October) seasons. The control sites were selected due to their proximity to the districts that had the outbreak; however, no YF cases had been recorded in these villages during the yellow fever outbreak.\u003c/p\u003e \u003cp\u003e \u003cb\u003eFigure 1: Map of Ghana indicating the study areas.\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eCharacterization of\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003ebreeding habitats and abundance of\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003elarvae\u003c/b\u003e\u003c/p\u003e \u003cp\u003eLarval sampling was performed in each study site, to characterise the breeding habitats and abundance of immature \u003cem\u003eAedes\u003c/em\u003e mosquitoes. During the larval surveys, the habitat type, its location in a household (indoor or outdoor), and its physical characteristics were recorded. Eight container types were classified based on their use and material: car tyres, discarded containers, drinking pots, drums, tanks, and buckets. Discarded containers and drinking pots were 50\u0026ndash;100 L capacity containers which included broken jars, bottles, small plastic food containers, tins, plates, cans, cooking pots, drinking troughs and broken pots made of clay, plastic or metal.\u003c/p\u003e \u003cp\u003eDrums were defined as 100-500L capacity plastic water storage containers. Tanks were 100\u0026ndash;500 L capacity water storage containers made of metal or concrete. Buckets included 10\u0026ndash;25 L water storage containers made of metal or plastic. In almost all the sites, access to pipe-borne water was a big challenge, therefore, households tend to store water in storage containers, pots, drums etc for long-term use. These containers may favour the breeding of the \u003cem\u003eAedes\u003c/em\u003e vector. Coordinates of all collection points were recorded using a GPSMAP\u0026reg; 60CSx geographical position system (GPS) instrument (Garmin International, Inc., Olathe, Kansas, USA).\u003c/p\u003e \u003cp\u003eAll larval samples were transported to the insectary at the Department of Medical Microbiology, University of Ghana, where they were raised to adults under suitable conditions (temperature: 27\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C, 75\u0026thinsp;\u0026plusmn;\u0026thinsp;10% relative humidity). The larvae were fed on TetraMin Baby fish food (Tetra Werke, Melle, Germany). Emerged adults were morphologically identified using standard taxonomic keys (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eFigure 2: Larval habitats encountered during larval sampling. a Drinking pot, b Tank, c Car tyre, g Discarded container, e Drum f Bucket\u003c/b\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eDetermination of Stegomyia Indices\u003c/h2\u003e \u003cp\u003eThe extent of infestation by \u003cem\u003eAedes\u003c/em\u003e mosquitoes was estimated using the classical Stegomyia indices including BI, Breteau Index, (the number of positive containers per 100 surveyed houses); HI; House Index, (percentage of houses positive for \u003cem\u003eAedes\u003c/em\u003e larvae or pupae); CI; Container Index, (the percentage of containers positive per 100 houses inspected). These larval and pupal indices continue to be the predominant and frequent measures used to evaluate vector prevalence because catching adult mosquitoes is time-consuming and requires access to private land (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e). Stegomyia indices are quantitative indicators of the risk of transmission.\u003c/p\u003e \u003cp\u003eUsing the WHO criteria, the risk of YF at each site was assessed as follows: In an area where BI, HI, and CI exceeded 50%, 35% and 20% respectively, the risk of \u003cem\u003eAe aegypti\u003c/em\u003e-transmitted Viral Haemorrhagic Fever (VHF) was considered to be high; an area where BI was between 5% and 50%, the density of \u003cem\u003eAe. aegypti\u003c/em\u003e was considered to be sufficient to promote an outbreak of VHF disease; an area where BI, HI and CI were less than 5%, 4% and 3% respectively, it was considered to be unlikely for YF transmission to occur (\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eAdult\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003emosquito sampling\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe spatio-temporal distribution of adult \u003cem\u003eAedes\u003c/em\u003e mosquitoes was determined by sampling indoors and outdoors using three methods; Biogent sentinel 2 traps (BG traps), Human landing catches (HLC), and prokopack aspiration (PPK) (John W. Hock Company, Gainesville, U.S.A.) (\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e). Geographical coordinates of each house sampled were taken. Cross-sectional surveys were undertaken in the dry season (March to May 2022) and in the rainy season (August to October 2022). Sixteen houses were randomly selected for each sampling method at each site (4 houses per day). Biogent sentinel traps were set both indoors (living rooms and bedrooms) and outdoors (open, verandas, or under a shed/tree where people sit to chat) about 5 m from the house) during the times 3:00\u0026ndash;7:00 pm. The BG traps were baited with carbon dioxide (CO\u003csub\u003e2\u003c/sub\u003e) which was produced from a mixture of 17.5 g yeast (Angel Yeast (Egypt) Co. Ltd.), 250 g sugar in 1 litre of water (van Loon \u003cem\u003eet al.\u003c/em\u003e, 2015). After the 4 hours, mosquitoes trapped were carefully removed, placed in a cooler box containing ice and then transported to the insectary. The HLC method was also used to sample host-seeking adult \u003cem\u003eAedes\u003c/em\u003e mosquitoes. On each day, one trained volunteer was positioned indoors and another outdoors and collected mosquitoes from 3:00\u0026ndash;6:00 pm. Collected \u003cem\u003eAedes\u003c/em\u003e were placed in well-labelled paper cups, and transported to the insectary for identification and further processing.\u003c/p\u003e \u003cp\u003eProkopack aspirations were used to sample resting mosquitoes. Sampling for \u003cem\u003eAedes\u003c/em\u003e mosquitoes was done indoors and outdoors. Sampled adult \u003cem\u003eAedes\u003c/em\u003e mosquitoes were knocked down with chloroform and preserved in Eppendorf tubes containing silica gel.\u003c/p\u003e \u003cp\u003e \u003cb\u003eMorphological identification of\u003c/b\u003e \u003cb\u003eA\u003c/b\u003e\u003cb\u003eedes mosquitoes\u003c/b\u003e\u003c/p\u003e \u003cp\u003eAll adult Aedes mosquitoes from the adult sampling, and those used for the susceptibility testing (Aedes larvae collected in the study sites and raised to adults), were identified morphologically using the taxonomic keys by Huang (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eInsecticide susceptibility tests\u003c/h2\u003e \u003cp\u003eInsecticide susceptibility test was conducted using WHO tubes to determine phenotypic resistance according to WHO criteria (\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e). Adult female Aedes mosquitoes that were 3\u0026ndash;5-day-old were exposed to papers impregnated with permethrin (0.75%), deltamethrin (0.05%), and pirimiphos-methyl (0.25%). Larvae and pupae collected from the larval sampling as well as the Ovitraps that were set were used for the WHO susceptibility testing. Though these doses are not the recommended doses for evaluating the susceptibility of \u003cem\u003eAedes\u003c/em\u003e mosquitoes, they are the most commonly used (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e). The knockdown time was recorded every 10 min during the 60-minute exposure period. Mortalities were recorded after a 24-hour recovery period. Alive (resistant) and dead (susceptible) mosquitoes were stored in absolute ethanol for later DNA analysis.\u003c/p\u003e \u003cp\u003e \u003cb\u003eGenotyping of kdr mutations in\u003c/b\u003e \u003cb\u003eAedes aegypti\u003c/b\u003e \u003cb\u003epopulations\u003c/b\u003e\u003c/p\u003e \u003cp\u003eA sub-sample of 242 phenotypically pyrethroid resistant and susceptible Aedes mosquitoes were genotyped of kdr mutations, F1534C, V1016I and V410L. Total DNA was extracted from whole mosquitoes using the DNeasy Tissue Kit (Qiagen, In USA). Genotyping of the kdr mutations was done using allele-specific multiplex PCR according to well-described protocols of Villanueva-Segura \u003cem\u003eet al.\u003c/em\u003e (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e)\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003e \u003cb\u003eDistribution and abundance of\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003elarval habitats.\u003c/b\u003e\u003c/p\u003e \u003cp\u003eA total of 535 larval habitats with 86 positive breeding habitats were surveyed from all the study sites, comprising six (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e) different habitat types over the entire sampling period. Overall, the most abundant habitat type was car tyres [59.3%, n\u0026thinsp;=\u0026thinsp;51/86], whereas the least were tanks [2.3%, n\u0026thinsp;=\u0026thinsp;2/86] and buckets [2.3%, n\u0026thinsp;=\u0026thinsp;2/86] (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). A significantly higher abundance of larval habitats was encountered during the rainy season [62.8%, n\u0026thinsp;=\u0026thinsp;54/86] than in the dry season [37.2%, n\u0026thinsp;=\u0026thinsp;32/86] (\u003cem\u003eX\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;18.5035, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;5, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.002). Within the 4 sites sampled, a significantly higher proportion (99.8%, 85/86) of larval habitats were encountered outdoors as compared to indoors (1.2%, 1/86) (\u003cem\u003eX\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;42.4941, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;5, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eLarval habitat distribution abundance per season and location.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eDry Season\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eRainy Season\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHabitat Type\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIndoor (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOutdoor (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eIndoor (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOutdoor (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTotal (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBucket\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1 (100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1 (1.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2 (2.3)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDiscarded container\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1 (3.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e10 (18.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e11 (12.8)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDrinking pot\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1 (3.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e14 (26.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e15 (17.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eDrum\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1 (3.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e4 (7.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5 (5.8)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTank\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1 (3.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1 (1.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2 (2.3)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCar Tyre\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e28 (87.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e23 (43.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e51 (59.3)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e0 (0.00)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e32 (37.21)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e1 (1.16)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e53 (61.63)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e86 (100)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eLarval densities of\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003eimmature\u003c/b\u003e\u003c/p\u003e \u003cp\u003eA total of 2,664 \u003cem\u003eAedes\u003c/em\u003e immatures were collected over the entire sampling period, of which the most productive habitat type was car tyres 65.1% (1,734/2664), whereas the least productive was tanks 1.1% (30/2664) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). \u003cem\u003eAedes\u003c/em\u003e larval abundance was slightly higher at 1,342 (50.4%) in the rainy season as compared to the dry season at 1,322 (49.6%) (\u003cem\u003eX\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;37.1991, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;28, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.115). During both seasons, the highest larval abundance was observed in Larabanga, a yellow fever outbreak site [n\u0026thinsp;=\u0026thinsp;1,472 (55.3%)], whereas the least was observed in Wenchi, also an outbreak site [n\u0026thinsp;=\u0026thinsp;217 (8. 1%)], Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. A significantly higher abundance of \u003cem\u003eAedes\u003c/em\u003e immatures were collected outdoors [n\u0026thinsp;=\u0026thinsp;2,626 (98.6%)] as compared to [n\u0026thinsp;=\u0026thinsp;38 (1.4%)] those collected indoors (\u003cem\u003eX\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;86.000, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;28, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe seasonal distribution of \u003cem\u003eAedes\u003c/em\u003e immatures across the study sites.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eWenchi\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eLarabanga\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eKpalsogu\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003ePagaza\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLocation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDry\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eRainy\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDry\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eRainy\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eDry\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eRainy\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eDry\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eRainy\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIndoor\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOutdoor\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e145\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e832\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e640\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e392\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e345\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eContainer Type\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBucket\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDiscarded container\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e122\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e101\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDrinking pot\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e210\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDrum\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTank\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCar Tyre\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e115\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e817\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e315\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e145\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e72\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e832\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e640\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e0\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e430\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e345\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e200\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eLarval Indices (Stegomyia Indices)\u003c/h2\u003e \u003cp\u003eLarval Indices calculated using the WHO formula for each study site were used to make inferences about the risk of transmission of yellow fever in the outbreak and non-outbreak zones studied. From the results obtained, in Larabanga the indices (BI: 84% CI:26.4%) exceeded the threshold and can be classified as a high-risk zone for yellow fever transmission as compared to all the other study areas, Wenchi (BI:42.3% CI:19.3% HI:23.1%), Kpalsogu (BI:37.7% CI:12.1% HI:6.7%) and Pagaza (BI: 30.2% CI:8.9% HI:9.4%) that had values within the range sufficient to promote an outbreak. These are shown in Fig.\u0026nbsp;3.\u003c/p\u003e \u003cp\u003e \u003cb\u003eFigure 3: Stegomyia Indices per study site\u003c/b\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eSpatio-temporal distribution of adult\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003emosquitoes\u003c/b\u003e\u003c/p\u003e \u003cp\u003eOverall, a total of 1,507 adult \u003cem\u003eAedes\u003c/em\u003e mosquitoes were collected from all the study areas, with \u003cem\u003eAedes aegypt\u003c/em\u003ei [92%, n\u0026thinsp;=\u0026thinsp;1386/1507] as the predominant species, followed by \u003cem\u003eAedes formosus\u003c/em\u003e [8%, n\u0026thinsp;=\u0026thinsp;121/1507]. A high abundance of \u003cem\u003eAedes\u003c/em\u003e mosquitoes was collected in Larabanga, a yellow fever outbreak site [\u003cem\u003eAedes aegypti\u003c/em\u003e 884/1386 (63.8%); \u003cem\u003eAedes formosus\u003c/em\u003e 112/121 (92.3%)], whereas the least abundance was collected in Pagaza, a non-yellow fever outbreak site [\u003cem\u003eAedes aegypti\u003c/em\u003e 75/1386 (5.4%); \u003cem\u003eAedes formosus\u003c/em\u003e 9/121 (7.7%)] (\u003cem\u003eX\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;52.061, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003eA high abundance of \u003cem\u003eAedes\u003c/em\u003e mosquitoes was sampled outdoors [n\u0026thinsp;=\u0026thinsp;914 (60.7%)] as compared to indoor collections [n\u0026thinsp;=\u0026thinsp;593 (39.3%); \u003cem\u003eX\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;68.38, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;1, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.01)]. A high abundance of adult \u003cem\u003eAedes\u003c/em\u003e mosquitoes was sampled for both indoor [n\u0026thinsp;=\u0026thinsp;342 (34.3%)] and outdoor [n\u0026thinsp;=\u0026thinsp;654 (65.7%)] collections in Larabanga whereas, the least recorded abundance was in Pagaza [Indoor (n\u0026thinsp;=\u0026thinsp;45 (53.6%); Outdoor (n\u0026thinsp;=\u0026thinsp;39, 46.4%; \u003cem\u003eX\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;32.021, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAdult \u003cem\u003eAedes\u003c/em\u003e abundance per study site and location\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eStudy Sites\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eAedes aegypti\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eAedes formosus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eIndoor\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eOutdoor\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eTotal per Site\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo. (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNo. (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNo. (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eNo. (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNo. (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWenchi\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e325 (100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0 (0.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e159 (48.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e166 (51.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e325 (21.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLarabanga\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e884 (88.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e112 (12.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e342 (34.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e654 (65.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e996 (66.6)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eKpalsogu\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e102 (100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0 (0.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e47 (46.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e55 (53.9)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e102 (6.7)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePagaza\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75 (89.0.)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9 (11.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e45 (53.6)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e39 (46.4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e84 (5.6)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e1386 (92.0)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e121 (8.0)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e593 (39.3)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e914 (60.7\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e1507 (100)\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eAdult \u003cem\u003eAedes\u003c/em\u003e mosquitoes were more predominant in the dry season [n\u0026thinsp;=\u0026thinsp;922/1507 (61.2%)] than in the rainy season [n\u0026thinsp;=\u0026thinsp;585/1507 (38.8%)], (\u003cem\u003eX\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;75.36, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;1, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). In the dry season, the highest abundance of adult \u003cem\u003eAedes\u003c/em\u003e mosquitoes was recorded in Larabanga [n\u0026thinsp;=\u0026thinsp;289/585, (49.4%)], followed by those from Wenchi [n\u0026thinsp;=\u0026thinsp;189/585, (32.3%)], both being yellow fever outbreak sites. Mosquitoes from the non-yellow fever outbreak sites were the least, Pagaza [n\u0026thinsp;=\u0026thinsp;70/585, (12.0%)] and Kpalsogu [n\u0026thinsp;=\u0026thinsp;37/585, (6.3%)]. Similarly, during the rainy season, the highest abundance of \u003cem\u003eAedes\u003c/em\u003e mosquitoes was recorded in Larabanga [n\u0026thinsp;=\u0026thinsp;707/922, (76.7%)], followed by Wenchi [n\u0026thinsp;=\u0026thinsp;136/922 (14.8%)], the yellow fever outbreak sites. There were small numbers of Aedes mosquitoes sampled in the non-yellow fever outbreak sites of Kpalsogu [n\u0026thinsp;=\u0026thinsp;65/922, (7.0%)] and Pagaza [n\u0026thinsp;=\u0026thinsp;14/922, (1.5%)]. These are shown in Fig.\u0026nbsp;4.\u003c/p\u003e \u003cp\u003e \u003cb\u003eFigure 4: Seasonal abundance of adult\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003emosquitoes\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003eSeasonal abundance of adult\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003emosquitoes per trap type\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThree separate traps namely, HLC, PPK, and BG were used to collect a total of 1507 adult Aedes mosquitoes from all the study sites. A total of 601 (39.9%) \u003cem\u003eAedes\u003c/em\u003e mosquitoes were collected using HLC [dry\u0026thinsp;=\u0026thinsp;280 (46.6%); rainy\u0026thinsp;=\u0026thinsp;321 (53.4%)], PPK 443 (29.4%) [dry\u0026thinsp;=\u0026thinsp;183 (41.3%); rainy\u0026thinsp;=\u0026thinsp;260 (58.7%)] and BG 463 (30.7%) [dry\u0026thinsp;=\u0026thinsp;122 (26.3%); rainy\u0026thinsp;=\u0026thinsp;341 (73.7%)]. Of the 796 female adult mosquitoes caught with PPK and BG, 575 [PPK\u0026thinsp;=\u0026thinsp;295 (66.6%); BG\u0026thinsp;=\u0026thinsp;280 (60.5%)] were unfed, 177 [PPK\u0026thinsp;=\u0026thinsp;77 (17.4%) BG\u0026thinsp;=\u0026thinsp;100 (21.6%)] were blood-fed, half-gravid [PPK\u0026thinsp;=\u0026thinsp;12 (2.7%); BG\u0026thinsp;=\u0026thinsp;0] and gravid [PPK\u0026thinsp;=\u0026thinsp;18 (4.1%); BG\u0026thinsp;=\u0026thinsp;14 (3.0%)], (\u003cem\u003eX\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;46.745, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;5).\u003c/p\u003e \u003cp\u003e \u003cb\u003eFigure 5: Season abundance of\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003emosquitoes per trap type (In-Indoor; OUT-Outdoor; BG-Biogents-2 sentinel traps; HLC-Human Landing Catches; PPK-Prokopack aspiration)\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003eInsecticide susceptibility status of\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003emosquitoes\u003c/b\u003e\u003c/p\u003e \u003cp\u003eBioassay results showed resistance to deltamethrin at x1 concentration across all the study sites (33.75% \u0026minus;\u0026thinsp;70%) (Fig.\u0026nbsp;6). However, susceptibility to deltamethrin at x5 was observed across all sites. The mosquitoes showed possible resistance to permethrin at x1 in Kpalsogu (95%), Pagaza (96.5%), and susceptibility in Larabanga (98%). Resistance to pirimiphos-methyl at x1 was shown in \u003cem\u003eAedes\u003c/em\u003e mosquitoes in Kpalsogu (63.75%) and possible resistance was observed in Pagaza (93.75%) and Larabanga (92.5%). For Wenchi, for logistical challenges, not many larvae were sampled for the determination of insecticide susceptibility, so only deltamethrin was run. These results are shown in Fig.\u0026nbsp;6.\u003c/p\u003e \u003cp\u003e \u003cb\u003eFigure 6: Mortalities of\u003c/b\u003e \u003cb\u003eAedes\u003c/b\u003e \u003cb\u003emosquitoes exposed to different insecticides in the study sites.\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cb\u003eGenotypic mutations associated with resistance in\u003c/b\u003e \u003cb\u003eAedes aegypti\u003c/b\u003e\u003c/p\u003e \u003cp\u003eA subset of 242 \u003cem\u003eAe. aegypti\u003c/em\u003e obtained from the phenotypic assays were genotyped for the F1534C, V1016I and V410L kdr mutations. About 20\u0026ndash;30 mosquito samples were selected from each site per insecticide paper per concentration for the genotypic resistance determination. The F1534C mutation was detected in moderate to high allele frequencies in \u003cem\u003eAe. aegypti\u003c/em\u003e mosquitoes exposed to pyrethroid insecticides. \u003cem\u003eAedes aegypti\u003c/em\u003e from Wenchi that were exposed to deltamethrin, had a significantly high allele frequency of F1534C mutation (0.92) compared to mosquitoes from the non-outbreak sites, Pagaza (0.19) and Kpalsogu (0.35) (\u003cem\u003eχ2\u003c/em\u003e\u0026thinsp;=\u0026thinsp;50.50, \u003cem\u003edf\u003c/em\u003e\u0026thinsp;=\u0026thinsp;3, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001). For the V1016I mutation, low to moderate allele frequencies (0.23 to 0.54) were observed in \u003cem\u003eAe. aegypti\u003c/em\u003e mosquitoes exposed to both deltamethrin and permethrin except for Larabanga, an outbreak site, where mosquitoes had a high allele frequency of 0.77. However, there was a significant difference in the frequency of V1016I mutation in outbreak and non-outbreak sites (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). For the V410L mutation, there was no significant differences in the frequency of the mutations between outbreak sites and non-outbreak sites with low allele frequencies ranging from 0.05 to 0.15. The genotypes and allele frequencies of each kdr mutation are shown in Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eNumber of genotypes and frequencies of the F1534C, V1016I and V410L mutation in the voltage-gated sodium channel gene of \u003cem\u003eAedes aegypti\u003c/em\u003e mosquitoes.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"16\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c16\" colnum=\"16\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eF1534C\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eV1016I\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c14\"\u003e \u003cp\u003eV410L\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eInsecticide\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSite Description\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStudy site\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eFF\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eFC\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAllele\u003c/p\u003e \u003cp\u003eFreq.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eII\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eVV\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eVI\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003eAllele\u003c/p\u003e \u003cp\u003eFreq.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003eLL\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c14\"\u003e \u003cp\u003eVV\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c15\"\u003e \u003cp\u003eVL\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c16\"\u003e \u003cp\u003eAllele\u003c/p\u003e \u003cp\u003eFreq.\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e\u003cb\u003eDeltamethrin\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003eNon-outbreak\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003ePagaza\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0.13\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eKpalsogu\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0.08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003eOutbreak\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eLarabanga\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e28\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eWenchi\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0.09\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e186\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e71\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e82\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e33\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e14\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e27\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003e145\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e6\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e\u003cb\u003e160\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003e22\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003e\u003cb\u003ePermethrin\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003eNon-outbreak\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003ePagaza\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0.15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eKpalsogu\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cb\u003eOutbreak\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eLarabanga\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eWenchi\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e56\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e13\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e\u003cb\u003e37\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e6\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e\u003cb\u003e13\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e\u003cb\u003e17\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c11\"\u003e \u003cp\u003e\u003cb\u003e26\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c13\"\u003e \u003cp\u003e\u003cb\u003e2\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c14\"\u003e \u003cp\u003e\u003cb\u003e53\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003e1\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c16\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\u003cp\u003eAbbreviations:\u0026nbsp;VV, wild type (susceptible); VL, heterozygotes; LL, mutant (resistant); VI, heterozygotes; II, mutant (resistant); FF, wild type (susceptible); FC, heterozygotes; CC, mutant (resistant); \u003cem\u003en\u003c/em\u003e sample size.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eDue to their capacity to spread a variety of arboviral infections like dengue, chikungunya, Zika, and yellow fever, Aedes mosquitoes pose an increasing hazard to public health on a global scale\u0026nbsp;(31).\u0026nbsp;Ghana is endemic for yellow fever with the most recent outbreak occurring in 2021\u0026nbsp;(13). Hence, the need to monitor the densities and insecticide susceptibility status of \u003cem\u003eAedes\u003c/em\u003e mosquitoes in the country. \u0026nbsp;This study sought to\u0026nbsp;determine the risk of arboviral transmission and insecticide resistance status of \u003cem\u003eAedes\u003c/em\u003e mosquitoes in a yellow fever outbreak and non-outbreak areas in Ghana. \u0026nbsp;From the data obtained, car tyres were the most productive habitats for \u003cem\u003eAedes\u003c/em\u003e mosquitoes seasonally. Moreover, the outbreak area (Larabanga) had a high Stegomyia indices value and hence satisfied the WHO criteria for a high-risk zone for yellow fever transmission. Adult \u003cem\u003eAedes aegypti\u003c/em\u003e was the predominant vector sampled in all the study sites, with high abundance in the dry season and from outdoor collection.\u0026nbsp;The distribution of larval habitat types varied significantly between seasons.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFindings from this study showed that car tyres were responsible for over 60% of \u003cem\u003eAedes\u003c/em\u003e immatures over the entire sampling period. Car tyres seems to provide an optimal temperature, humidity and sufficient light intensity to ensure larvae development\u0026nbsp;(32).\u0026nbsp;This finding is consistent with studies by Owusu-Asenso \u003cem\u003eet al.\u003c/em\u003e (11)\u0026nbsp;in Ghana and Kamgang \u003cem\u003eet al.\u003c/em\u003e(33)\u0026nbsp;in the\u0026nbsp;Central African Republic who also found car tyres as the most conducive and productive breeding habitat for \u003cem\u003eAedes aegypti\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eIn the current study, results from the larval indices indicate that Larabanga had values that exceeded the WHO threshold and hence is a high-risk zone for arboviral pathogen transmission. This is similar to a previous study by Appawu \u003cem\u003eet al\u003c/em\u003e.\u0026nbsp;(10)\u0026nbsp;in Larabanga, where the Stegomyia indices exceeded the threshold and was considered as a high-risk zone for yellow fever transmission. Furthermore, Stegomyia indices values within the WHO criteria reported in Wenchi, Kpalsogu and Pagaza were sufficient to promote an outbreak within these sites. These findings imply that inhabitants within these study sites are at risk of yellow fever infection, Hence, there is a need to employ preventive measures through vaccination and effective vector control strategies.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSeasonal variation in population density is common seasonally for \u003cem\u003eAedes\u003c/em\u003e mosquitoes due to their sensitivity to changes in temperature and rainfall (34).\u0026nbsp;This study found a significantly higher abundance of \u003cem\u003eAedes\u003c/em\u003e immature during the rainy season.\u0026nbsp;The development of mosquitoes, their survival and the effective transmission of pathogens are influenced by humidity, temperature and rainfall\u0026nbsp;(35,36).\u0026nbsp;Sufficient humidity and rainfall influence the breeding sites, increasing vector populations\u0026nbsp;(37). Hence, an increase in breeding sites may explain the observed increase in \u003cem\u003eAedes\u003c/em\u003e immatures in the rainy season.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThis finding corroborates with studies in Ghana by Owusu-Asenso et al.\u0026nbsp;(11)\u0026nbsp;and in Kenya\u0026nbsp;by\u0026nbsp;Ndenga \u003cem\u003eet al.\u003c/em\u003e (38). Their findings showed high densities of \u003cem\u003eAedes\u003c/em\u003e immatures in the rainy season. However, this finding was contrary to another study in Ghana where \u003cem\u003eAedes\u003c/em\u003e larvae were found predominantly in the dry season. He reported that during drought conditions, the surge in the storage of water creates more breeding habitats for \u003cem\u003eAedes\u0026nbsp;\u003c/em\u003emosquitoes, causing an increase in their abundance\u0026nbsp;(10).\u003c/p\u003e\n\u003cp\u003eIt was observed that adult\u0026nbsp;\u003cem\u003eAedes\u003c/em\u003e mosquitoes were\u0026nbsp;predominantly\u0026nbsp;collected during the dry season and in outdoor settings. Warmer temperatures enhance the growth of mosquitoes and viral replication\u0026nbsp;(34). The life-limiting elements of latitude, altitude, temperature, rainfall, humidity, season, habitat, and dispersal have an impact on the distribution and population of Aedes\u0026nbsp;mosquitoes\u0026nbsp;(10).\u0026nbsp;Finding more adult \u003cem\u003eAedes\u003c/em\u003e mosquitoes outside may suggest their exophilic nature as reported in previous studies in Ghana and Kenya\u0026nbsp;(11,39). It is important to note that people spending more time outdoors compared to indoors influences the biting and feeding behaviour of \u003cem\u003eAedes\u003c/em\u003e mosquitoes\u0026nbsp;(38).\u0026nbsp;Studies in Ghana have also suggested that \u003cem\u003eAedes\u003c/em\u003e mosquitoes often rest outdoors before and after blood feeding\u0026nbsp;(40)\u0026nbsp;and were more abundant from outdoor collection in the dry season\u0026nbsp;(10).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIt was observed in this study that the majority of the adult \u003cem\u003eAedes\u003c/em\u003e mosquitoes were \u003cem\u003eAedes aegypti\u0026nbsp;\u003c/em\u003ewhich is responsible for yellow fever transmission in Ghana and can transmit other arboviral pathogens such as dengue fever virus\u0026nbsp;(12). There have been previous reports of dengue viral infections in children in Ghana\u0026nbsp;(14)\u0026nbsp;and exposure to dengue and chikungunya\u0026nbsp;(15,16,41)\u0026nbsp;that show the role of \u003cem\u003eAedes aegypti\u003c/em\u003e in the transmission of multiple arboviral pathogens in Ghana, which cannot be overlooked.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn this study, \u003cem\u003eAedes\u003c/em\u003e mosquitoes\u0026nbsp;across the\u0026nbsp;sites\u0026nbsp;showed\u0026nbsp;resistance to Deltamethrin. This\u0026nbsp;might\u0026nbsp;be due to the indirect impact of the use of insecticides for public health vector control such as the use of Long-Lasting Insecticidal Nets (LLINs) and IRS, as well as pesticide use in agriculture\u0026nbsp;(11,42,43). This finding is similar to that reported in a study conducted in Ghana (Abdulai \u003cem\u003eet al.,\u003c/em\u003e 2023). Whereas \u003cem\u003eAedes\u003c/em\u003e mosquitoes collected from Pagaza and Kpalsogu showed suspected resistance to Permethrin, samples from Larabanga were found to be susceptible to Permethrin. Similarly, pyrethroid resistance has also been reported in \u003cem\u003eAedes\u0026nbsp;\u003c/em\u003e\u003cem\u003eaegypti\u0026nbsp;\u003c/em\u003epopulations from Ghana and other West African countries ((11,17,18,29)\u0026nbsp;\u003cem\u003eAedes\u003c/em\u003e mosquitoes in this study were also resistant or possibly resistant to Pirimiphos-methyl. The findings suggest that resistance to this organophosphate by the \u003cem\u003eAedes\u003c/em\u003e population has increased.\u003c/p\u003e\n\u003cp\u003eIn this study, the F1534C and V1016I kdr mutations were high in frequencies in resistant and susceptible \u003cem\u003eAedes\u003c/em\u003e mosquitoes while V410L kdr mutation showed low frequencies. Although suspected resistance and susceptibility to permethrin were recorded in the \u003cem\u003eAedes\u003c/em\u003e mosquitoes from the same population, there is the risk of resistance developing over time due to the high frequencies of the F1534C and V1016I kdr mutations. Pyrethroid resistance in \u003cem\u003eAedes aegypti\u003c/em\u003e is a worldwide challenge for mosquito control due to its use for insecticide-treated nets and indoor residual spraying\u0026nbsp;(44). Similarly, other studies in Ghana have found high frequencies of V1016I\u0026nbsp;and\u0026nbsp;F1534C kdr mutations in both pyrethroid-susceptible and resistant \u003cem\u003eAedes\u003c/em\u003e mosquitoes collected in Ghana\u0026nbsp;(9,21). This suggests that other resistance mechanism such as metabolic resistance may be involved in insecticide resistance of \u003cem\u003eAedes aegypti\u003c/em\u003e populations in Ghana. Hence, further studies are needed to elucidate the mechanisms mediating insecticide resistance in Aedes mosquitoes in Ghana.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eOur findings reveal that \u003cem\u003eAedes\u003c/em\u003e immatures were most abundant in the dry season and car tyres were the most productive habitat type. \u003cem\u003eAedes aegypti\u003c/em\u003e were the predominant species found.\u0026nbsp;The results showed that Larabanga, the epicentre of the yellow fever outbreak is a high-risk zone for arboviral pathogen transmission. In addition,\u0026nbsp;high phenotypic and genotypic\u0026nbsp;resistance was observed in Aedes mosquito populations in Ghana.\u0026nbsp;Surveillance of \u003cem\u003eAedes\u003c/em\u003e mosquito bionomics and insecticide susceptibility in Ghana is crucial to help in the development of arboviral vector control strategies to control and prevent arboviral outbreaks in Ghana.\u0026nbsp;\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003e\u003cem\u003eAe\u003c/em\u003e.: \u003cem\u003eAedes\u003c/em\u003e; DNA: Deoxyribonucleic acid; PCR: polymerase chain reaction; PMI: President Malaria Initiative; WHO: World Health Organization; YF: Yellow Fever; HLC: Human Landing Catch; PPK: Procopack; BG: Biogent sentinel trap; GHS: Ghana Health Service; HI: House Index; CI: Container Index; BI: Breateau Index; VHF: Viral Haemorrhagic Fever; GPS: Geographical position system\u0026nbsp;\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWeappreciate the management and staff of PMI-Vector Link Tamale, especially Mr Osei Akuoko, Mr Imoro Nurudeen, Mr Cosmos Pambit Zong and Mr Abdullai Abdul-Rahman for granting permission to use their facility and resources to carry out this project. Special thanks to the Centre for Vector-borne Disease Research (CVBDR) team\u0026nbsp;at the Department of Medical Microbiology, University of Ghana Medical School, for the support and technical advice during this project. We also appreciate all the indigenes in the various study sites who offered field assistance.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by grants from the National Institute of Health (D43 TW 011513).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll the data supporting this study are included in the article.\u003c/p\u003e\n\u003cp\u003eEthical Consideration\u003c/p\u003e\n\u003cp\u003eEthical approval for this study was obtained from the University of Ghana, College of Health Sciences (CHS) Ethics and Protocol Review Committee (EPRC) (CHS-Et/M.11- P4.1/2021-2022).\u0026nbsp;Meetings were held in each study site with chiefs and the residents to introduce the research study to the public. All study participants recruited for mosquito collection in this study were adults (18 years old and above). A written and signed consent was obtained from all adults who volunteered to participate in HLC. They were then trained before sampling of mosquitoes. A copy of the signed consent form was given to each of the HLC volunteers and another was kept in a locked cabinet with restricted access in offices at The Department of Medical Microbiology, University of Ghana Medical School. Verbal consent was obtained from household heads to sample mosquitoes in their houses and compounds.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eMOA, SKA and YAA were responsible for the study design, supervised the data collection, and contributed to the writing of the manuscript. MOA, IKS, AA, CMO-A, ED and ARM carried out the data collection, laboratory work and data analysis. MOA, CMO-A and AA drafted and revised the manuscript. All authors read and approved the final copy of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eWHO. Geneva, World Health Organisation. 2016 [cited 2022 Jul 20]. Monitoring and managing insecticide resistance in Aedes mosquito populations, interim guidance for entomologists. Available from: https://www.who.int/csr/resources/publications/zika/insecticide-resistance/en/\u003c/li\u003e\n\u003cli\u003eGardner CL, Ryman KD. Yellow fever: a reemerging threat. (Special Issue: Emerging pathogens. Clin Lab Med. 2010;30(1):237\u0026ndash;60. \u003c/li\u003e\n\u003cli\u003eSilva NIO, Sacchetto L, De Rezende IM, Trindade GDS, Labeaud AD, De Thoisy B, et al. Recent sylvatic yellow fever virus transmission in Brazil: The news from an old disease. Vol. 17, Virology Journal. 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Evidence of chikungunya virus infections among febrile patients at three secondary health facilities in the Ashanti and the Bono Regions of Ghana. 2021 [cited 2024 Apr 1]; Available from: https://doi.org/10.1371/journal.pntd.0009735\u003c/li\u003e\n\u003cli\u003eKristan M, Fleischmann H, Della Torre A, Stich A, Curtis CF. Pyrethroid resistance/susceptibility and differential urban/rural distribution of Anopheles arabiensis and An.\u0026emsp;gambiae s.s. malaria vectors in Nigeria and Ghana. Med Vet Entomol [Internet]. 2003 Sep 1;17(3):326\u0026ndash;32. Available from: https://doi.org/10.1046/j.1365-2915.2003.00449.x\u003c/li\u003e\n\u003cli\u003eAdutwum A, 1* K, Mensah BA, Agyemang TK. Characterization of mosquito larval habitats and assessment of insecticide‐resistance status of Anopheles gambiae senso lato in urban areas in southwestern Ghana. Wiley Online LibraryAA Kudom, BA Mensah, TK AgyemangJournal of Vector Ecology, 2012\u0026bull;Wiley Online Library [Internet]. 2012 Jun [cited 2024 Apr 1];37(1):77\u0026ndash;82. Available from: https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1948-7134.2012.00202.x\u003c/li\u003e\n\u003cli\u003eMoyes CL, Vontas J, Martins AJ, Ng LC, Koou SY, Dusfour I, et al. Contemporary status of insecticide resistance in the major Aedes vectors of arboviruses infecting humans. PLoS Negl Trop Dis [Internet]. 2017 Jul 20;11(7):e0005625\u0026ndash;e0005625. Available from: https://pubmed.ncbi.nlm.nih.gov/28727779\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"parasites-and-vectors","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"parv","sideBox":"Learn more about [Parasites \u0026 Vectors](http://parasitesandvectors.biomedcentral.com/)","snPcode":"13071","submissionUrl":"https://submission.nature.com/new-submission/13071/3","title":"Parasites \u0026 Vectors","twitterHandle":"@bugbittentweets","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Aedes aegypti, yellow fever, Stegomyia indices, Ghana","lastPublishedDoi":"10.21203/rs.3.rs-4271509/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4271509/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn late 2021, Ghana was hit by a Yellow Fever outbreak that started in two (2) districts in the Savannah region and spread to several other Districts in (3) regions (Oti, Bono and Upper West).\u003c/p\u003e\n\u003cp\u003eYellow fever is endemic in Ghana. However, there is currently no structured vector control programme for the yellow vector, Aedes mosquitoes in Ghana. Knowledge of \u003cem\u003eAedes \u003c/em\u003ebionomics and insecticide susceptibility status is important to control the vectors. This study therefore sought todetermine \u003cem\u003eAedes \u003c/em\u003evector bionomics and their insecticide resistance status during a yellow fever outbreak.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was performed in two yellow fever outbreak sites (Wenchi, Larabanga) and two non-outbreak sites (Kpalsogu, Pagaza) in Ghana. Immature \u003cem\u003eAedes\u003c/em\u003e mosquitoes were sampled from water-holding containers in and around human habitations. The risk of disease transmission was determined in each site using stegomyia indices. Adult \u003cem\u003eAedes\u003c/em\u003e mosquitoes were sampled using Biogents Sentinel (BG) traps, Human Landing Catch (HLC), and Prokopack (PPK) aspirators. Phenotypic resistance was determined with WHO susceptibility tests using \u003cem\u003eAedes \u003c/em\u003emosquitoes collected as larvae and reared into adults. Knockdown resistance (kdr) mutations were detected using allele-specific multiplex PCR.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eOf the 2,664 immature \u003cem\u003eAedes\u003c/em\u003e sampled, more than 60% were found in car tyres. Larabanga, an outbreak site, was classified as a high-risk zone for the Yellow Fever outbreak (BI: 84%, CI: 26.4%). Out of 1,507 adult\u003cem\u003e Aedes \u003c/em\u003emosquitoes collected, \u003cem\u003eAedes aegypti\u003c/em\u003e was the predominant vector species (92%). A significantly high abundance of \u003cem\u003eAedes\u003c/em\u003e mosquitoes was observed during the dry season (61.2%) and outdoors (60.6%) (\u003cem\u003eP \u003c/em\u003e\u0026lt; 0.001). Moderate to high resistance to deltamethrin was observed in all sites (33.75% to 70%). Moderate resistance to pirimiphos-methyl (65%) was observed in Kpalsogu. \u003cem\u003eAedes\u003c/em\u003emosquitoes from Larabanga were susceptible (98%) to permethrin. The F1534C kdr, V1016I kdr and V410 kdr alleles were present in all the sites with frequencies between (0.05-0.92). The outbreak sites had significantly higher allele frequencies of F1534C and V1016I respectively compared to non-outbreak sites (\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.001).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study indicates that \u003cem\u003eAedes\u003c/em\u003e mosquitoes in Ghana pose a significant risk to public health, and there is a need for continuous surveillance to inform effective vector control strategies.\u003c/p\u003e","manuscriptTitle":"Risk of Arboviral Transmission and Insecticide Resistance Status of Aedes Mosquitoes during a Yellow Fever Outbreak in Ghana","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-19 06:56:20","doi":"10.21203/rs.3.rs-4271509/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorAssigned","content":"","date":"2024-04-16T10:06:00+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-04-16T09:55:11+00:00","index":"","fulltext":""},{"type":"submitted","content":"Parasites \u0026 Vectors","date":"2024-04-15T18:49:56+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"parasites-and-vectors","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"parv","sideBox":"Learn more about [Parasites \u0026 Vectors](http://parasitesandvectors.biomedcentral.com/)","snPcode":"13071","submissionUrl":"https://submission.nature.com/new-submission/13071/3","title":"Parasites \u0026 Vectors","twitterHandle":"@bugbittentweets","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"60e693de-2475-4691-9bb3-50430b63c755","owner":[],"postedDate":"April 19th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2024-04-19T06:56:20+00:00","versionOfRecord":[],"versionCreatedAt":"2024-04-19 06:56:20","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4271509","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4271509","identity":"rs-4271509","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}