Risk of arboviral transmission and insecticide resistance status of Aedes mosquitoes during a yellow fever outbreak in Ghana.

BACKGROUND: In late 2021, Ghana was hit by a Yellow Fever outbreak that started in two districts in the Savannah region and spread to several other Districts in three regions. Yellow fever is endemic in Ghana. However, there is currently no structured vector control programme for Aedes the arboviral vector in Ghana. Knowledge of Aedes bionomics and insecticide susceptibility status is important to control the vectors. This study therefore sought to determine 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 to permethrin, deltamethrin and pirimiphos-methyl 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: Among 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. Hence there is a need to continue monitoring these vectors to develop an effective control strategy.

Larval habitat stability and productivity in two sites in Southern Ghana.

BACKGROUND: Mosquito larval source management (LSM) is a valuable additional tool for malaria vector control. Understanding the characteristics of mosquito larval habitats and its ecology in different land use types can give valuable insight for an effective larval control strategy. This study determined the stability and productivity of potential anopheline larval habitats in two different ecological sites: Anyakpor and Dodowa in southern Ghana. METHODS: A total of 59 aquatic habitats positive for anopheline larvae were identified, and sampled every two weeks for a period of 30 weeks using a standard dipping method. Larvae were collected using standard dippers and were raised in the insectary for identification. Sibling species of the Anopheles gambiae sensu lato (s.l.) were further identified by polymerase chain reaction. The presence of larval habitats, their stability and larvae positive habitats were compared between the two sites using Mann-Whitney U and the Kruskal-Wallis test. Factors affecting the presence of An. gambiae larvae and physicochemical properties at the sites were determined using multiple logistic regression analysis and Spearman's correlation. RESULTS: Out of a total of 13,681 mosquito immatures collected, 22.6% (3095) were anophelines and 77.38% (10,586) were culicines. Out of the 3095 anophelines collected, An. gambiae s.l. was predominant (99.48%, n = 3079), followed by Anopheles rufipes (0.45%, n = 14), and Anopheles pharoensis (0.064%, n = 2). Sibling species of the An. gambiae consisted of Anopheles coluzzii (71%), followed by An. gambiae s.s. (23%), and Anopheles melas (6%). Anopheles mean larval density was highest in wells [6.44 (95% CI 5.0-8.31) larvae/dip], lowest in furrows [4.18 (95% CI 2.75-6.36) larvae/dip] and man-made ponds [1.20 (95% CI 0.671-2.131) larvae/dip].The results also revealed habitat stability was highly dependent on rainfall intensity, and Anopheles larval densities were also dependent on elevated levels of pH, conductivity and TDS. CONCLUSION: The presence of larvae in the habitats was dependent on rainfall intensity and proximity to human settlements. To optimize the vector control measures of malaria interventions in southern Ghana, larval control should be focused on larval habitats that are fed by underground water, as these are more productive habitats.

Perceptions of the roles, impact, challenges and needs of community drug distributors in the control and elimination of neglected tropical diseases in difficult-to-access communities in Ghana.

INTRODUCTION: The success of mass drug administration (MDA) campaigns to control and eliminate neglected tropical diseases (NTDs) in Ghana depends, to a large extent, on the essential role community drug distributors (CDDs) play. This study aimed to investigate community's perceptions of CDDs' roles, impact of CDDs' work, challenges faced by CDDs, and views on resources required to enhance CDDs' work to sustain MDA campaigns. METHODS: A cross-sectional qualitative study employing the use of focus group discussions (FGDs) with community members and CDDs in selected NTD endemic communities together with individual interviews with district health officers (DHOs) was conducted. We interviewed 104 people aged 18 and over, purposively selected, through eight individual interviews, and 16 focus group discussions. RESULTS: Participants in the community FGDs noted that health education and the distribution of drugs were the main roles of CDDs. Participants also perceived that the work of CDDs had prevented the onset of NTDs, treated symptoms of NTDs, and generally reduced the incidence of infections. In the interviews with CDDs and DHOs, lack of cooperation/non-compliance by community members, demands by community members, lack of working resources and low financial motivation were mentioned as the main challenges to the work of CDDs. Moreover, the provision of logistics and financial motivation for CDDs were identified as factors that will enhance their work. CONCLUSIONS: Incorporating more attractive schemes will incentivise CDDs to improve output. Addressing the challenges highlighted is an important step for the work of CDDS to be effective in controlling NTDs in difficult-to-access communities in Ghana.

Coverage of preventive measures and surveillance for neglected tropical diseases in hard-to-reach communities in Ghana.

BACKGROUND: Neglected tropical diseases (NTDs) are a major public health burden which mainly affects poor populations living in tropical environments and hard-to-reach areas. The study sought to examine coverage of preventive efforts, and case surveillance for NTDs in hard-to-reach communities in Ghana. METHODS: The study investigated treatment efforts for lymphatic filariasis (LF), and onchocerciasis and schistosomiasis/soil transmitted helminths (SCH/STH) at household level, in difficult-to-access communities in Ghana. A total of 621 households were sampled from 6 communities in the Western, Oti and Greater Accra regions. RESULTS: Over 95% of the households surveyed were covered under mass drug administration (MDA) campaigns for lymphatic filariasis (LF) and onchocerciasis. More than 80% of households had received at least two visits by community drug distributors under the MDA campaigns in the last two years preceding the study. In addition, over 90% of households in the LF and onchocerciasis endemic communities had at least one member using anthelminthic medications under the MDA campaigns in the 12 months preceding the study. However, households where no member had taken anthelminthic medications in 12 months preceding the study were over 6 times likely to have someone in the household with LF. CONCLUSIONS: This study determined that SCH/STH, LF and onchocerciasis are of serious public health concern in some communities in Ghana. There is an urgent need for holistic practical disease control plan involving both financial and community support to ensure total control of NTDs in difficult-to-access communities is achieved.

Larval habitat diversity, physicochemical characterization and their effect on the density of larval malaria vectors in the city of Accra, Ghana.

Background: Malaria is more prevalent in rural areas due to fewer mosquito breeding habitats in urban settings. However, urban factors such as irrigated farming, open sewers, and discarded containers create mosquito breeding sites. This study investigates the diversity and distribution of larval habitats and the impact of physicochemical characteristics on the presence and density of Anopheles gambiae s.l. larvae in Accra, Ghana. Methods: Larval surveys and collections were conducted at fifteen locations in Accra, divided into five categories: Irrigated Urban Farming (IUF), Lower Socioeconomic Status (LS), Middle Socioeconomic Status (MS), High Socioeconomic Status (HS), and Peri-urban (PU) areas. Physicochemical parameters were measured, and species identification was performed using morphological and molecular methods. Results: A total of 727 breeding habitats were identified, with 65.34% (475/727) positive for Anopheles larvae. Drainage ditches were the most common habitat type (48.21%, 229/475). The highest abundance of An. gambiae s.l. was found in IUF sites (6,244/22,919), especially during the rainy season (77.01%, 17,650/22,919; R2 = 3.46, P = 0.000). Polluted habitats, including household effluents, had higher ammonium levels (3.4 mg/L NH -N) compared to unpolluted ones (1.3 mg/L NH -N). Other distinguishing parameters included dissolved oxygen (34% vs 52.9%), conductivity (5106 µS/cm vs 2049 µS/cm), and total dissolved solids (3181 mg/L vs 1255 mg/L). The predominant malaria vector was An. coluzzii(54.4%, 368/677). Additionally, the invasive An. stephensi, previously unreported in Ghana, was detected. Conclusion: Malaria vectors breed in diverse and often polluted urban habitats, with high larval densities in urban agricultural areas. The detection of the invasive An. Stephensi highlights the need for continuous monitoring and vector control strategies in urban settings.

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

Background: In 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).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. Aedesmosquitoes 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.

The role of car tyres in the ecology of Aedes aegypti mosquitoes in Ghana.

Aedes aegypti is an important vector of arboviral diseases including dengue and yellow fever. Despite the wide distribution of this mosquito species, there are limited data on the ecology of Ae. aegypti in Ghana. In this study, we report on the oviposition preference and the larval life tables of Ae. aegypti mosquitoes in Accra, Ghana. The oviposition preference of the mosquitoes to three habitat types (car tyres, drums and bowls) was measured by setting up ovitraps. We recorded the presence and abundance of larvae every 3 days. Two-hour-old Ae. aegypti larvae were introduced and raised in three habitat types to undertake larval life tables. The number of surviving larvae at each developmental stage was recorded daily until they emerged as adults. Car tyres showed a higher abundance of Ae. aegypti larvae (52.3%) than drums (32.5%) and bowls (15.1%) (ANOVA, F(2,159) = 18.79, P < 0.001). The mean development time of Ae. aegypti larvae was significantly lower in car tyres (7 ± 1 days) compared to that of bowls (9 ± 0.0 days) and drums (12.6 ± 1.5 days) (P = 0.024). The differences in pupation rates and emergence rates were not significant across the habitat types; however, the highest pupation rate was observed in bowls (0.92 ± 0.17) and the emergence rate was highest in tyres (0.84 ± 0.10). The proportion of first-instar larvae that survived to emergence was significantly higher in car tyres (0.84 ± 0.10) compared to that of bowls (0.72 ± 0.20) and drums (0.62 ± 0.20) (P = 0.009). No mortalities were observed after 9 days in car tyres, 10 days in bowls and 15 days in drums. The results confirm that discarded car tyres were the preferred habitat choice for the oviposition of gravid female Ae. aegypti mosquitoes and provide the best habitat conditions for larval development and survival. These findings are necessary for understanding the ecology of Ae. aegypti to develop appropriate strategies for their control in Ghana.

The Role of Car Tyres in the Ecology of Aedes aegypti Mosquitoes in Ghana.

Background: The Aedes aegyptimosquito is an important vector of arboviral diseases including dengue and yellow fever. Despite the wide distribution of the Aedes aegypti mosquito, there is limited data on the ecology of Aedes aegypti mosquitoes in Ghana. In this study, we report on the oviposition preference and the larval life table of Aedes aegypti mosquitoes in Accra, Ghana. Methods: The oviposition preference of Aedesmosquitoes to three habitat types (tyres, drums and bowls) was measured by setting up ovitraps. Ovitraps were checked for the presence of Aedes larvae every 3 days. The presence and number of larvae were recorded for each habitat type. Two-hour-old Aedes aegypti larvae were introduced into and raised in these three habitat types to undertake larval life tables. The number of surviving larvae at each developmental stage was recorded daily until they emerge as adults. Results: Car tyres showed a high abundance of Aedeslarvae (52.33%) than drums (32.49%) and bowls (15.18%) (ANOVA, F _ 18.79, df _ 2, 159, P < 0.001). The mean development time of Ae. aegypti larvae was significantly lower in car tyres (7 ± 1 days) compared to that of bowls (9 ± 0.0 days) and drums (12.6 ± 1.5 days) (H (2) = 7.448, P = 0.024). The differences in pupation rates and emergence rates were not significant across the habitat types, however, the highest pupation rate was observed in bowls (0.92) and the emergence rate was highest in tyres (0.84). The proportion of first-instar larvae that survived to adults was significantly higher in tyres with a shorter survival time (0.84; 9 days) compared to that of bowls (0.72; 10 days) and drums (0.62 ± 0.2; 13 days) (H (2) = 2.822, P= 0.009). Conclusion: The results confirm that discarded car tyres were the preferred habitat choice for the oviposition of gravid female Aedes aegypti mosquitoes and provide the best habitat condition for larval development and survival. These findings are necessary for understanding the ecology of Aedes mosquitoes to develop appropriate strategies for their control in Ghana.

Perceptions of the roles, impact, challenges and needs of community drug distributors in the control and elimination of neglected tropical diseases in difficult-to-access communities in Ghana.

The success of mass drug administration (MDA) campaigns to control and eliminate neglected tropical diseases (NTDs) in Ghana depends, to a large extent, on the essential role community drug distributors (CDDs) play. This study aimed to investigate community's perceptions of CDDs' roles, impact of CDDs' work, challenges faced by CDDs and views on resources required to enhance CDDs' work to sustain MDA campaigns. Methods : A cross-sectional qualitative study employing the use of focus group discussions (FGDs) with community members and CDDs in selected NTD endemic communities together with individual interviews with district health officers (DHOs)was conducted. We interviewed 104 people aged 18 and over, purposively selected, through eight individual interviews, and 16 focus group discussions. Results : Participants in the community FGDs noted that health education and distribution of drugs were the main roles of CDDs. Participants also perceived that the work of CDDs had prevented the onset of NTDs, treated symptoms of NTDs and generally reduced the incidence of infections. In the interviews with CDDs and DHOs, lack of cooperation/non-compliance by community members, demands by community members, lack of working resources and low financial motivation were mentioned as the main challenges to the work of CDDs. Moreover, provision of logistics and financial motivation for CDDs were identified as factors that will enhance their work. Conclusions : Incorporating more attractive schemes shall incentivise CDDs to improve output. Addressing the challenges highlighted is an important step for the work of CDDS to be effective in controlling NTDs in difficult-to-access communities in Ghana.

Insecticide resistance status of Aedes aegypti in southern and northern Ghana.

BACKGROUND: Outbreaks of Aedes-borne arboviral diseases are becoming rampant in Africa. In Ghana, there is no organized arboviral control programme with interventions restricted to mitigate outbreaks. Insecticide application is a crucial part of outbreak responses and future preventative control measures. Thus, knowledge of the resistance status and underlying mechanisms of Aedes populations is required to ensure optimal insecticide choices. The present study assessed the insecticide resistance status of Aedes aegypti populations from southern Ghana (Accra, Tema and Ada Foah) and northern Ghana (Navrongo) respectively. METHODS: Phenotypic resistance was determined with WHO susceptibility tests using Ae. aegypti collected as larvae and reared into adults. Knockdown resistance (kdr) mutations were detected using allele-specific PCR. Synergist assays were performed with piperonyl butoxide (PBO) to investigate the possible involvement of metabolic mechanisms in resistance phenotypes. RESULTS: Resistance to DDT was moderate to high across sites (11.3 to 75.8%) and, for the pyrethroids deltamethrin and permethrin, moderate resistance was detected (62.5 to 88.8%). The 1534C kdr and 1016I kdr alleles were common in all sites (0.65 to 1) and may be on a trajectory toward fixation. In addition, a third kdr mutant, V410L, was detected at lower frequencies (0.03 to 0.31). Pre-exposure to PBO significantly increased the susceptibility of Ae. aegypti to deltamethrin and permethrin (P < 0.001). This indicates that in addition to kdr mutants, metabolic enzymes (monooxygenases) may be involved in the resistance phenotypes observed in the Ae. aegypti populations in these sites. CONCLUSION: Insecticide resistance underpinned by multiple mechanisms in Ae. aegypti indicates the need for surveillance to assist in developing appropriate vector control strategies for arboviral disease control in Ghana.

The resting behavior of malaria vectors in different ecological zones of Ghana and its implications for vector control.

BACKGROUND: In sub-Saharan Africa there is widespread use of long-lasting insecticidal nets and indoor residual spraying to help control the densities of malaria vectors and decrease the incidence of malaria. This study was carried out to investigate the resting behavior, host preference and infection with Plasmodium falciparum of malaria vectors in Ghana in the context of the increasing insecticide resistance of malaria vectors in sub-Saharan Africa. METHODS: Indoor and outdoor resting anopheline mosquitoes were sampled during the dry and rainy seasons in five sites in three ecological zones [Sahel savannah (Kpalsogo, Pagaza, Libga); coastal savannah (Anyakpor); and forest (Konongo)]. Polymerase chain reaction-based molecular diagnostics were used to determine speciation, genotypes for knockdown resistance mutations (L1014S and L1014F) and the G119S ace1 mutation, specific host blood meal origins and sporozoite infection in the field-collected mosquitoes. RESULTS: Anopheles gambiae sensu lato (s.l.) predominated (89.95%, n = 1718), followed by Anopheles rufipes (8.48%, n = 162) and Anopheles funestus s.l. (1.57%, n = 30). Sibling species of the Anopheles gambiae s.l. revealed Anopheles coluzzii accounted for 63% (95% confidence interval = 57.10-68.91) and 27% (95% confidence interval = 21.66-32.55) was Anopheles gambiae s. s.. The mean resting density of An. gambiae s.l. was higher outdoors (79.63%; 1368/1718) than indoors (20.37%; 350/1718) (Wilcoxon rank sum test, Z = - 4.815, P < 0.0001). The kdr west L1014F and the ace1 mutation frequencies were higher in indoor resting An. coluzzii and An. gambiae in the Sahel savannah sites than in the forest and coastal savannah sites. Overall, the blood meal analyses revealed that a larger proportion of the malaria vectors preferred feeding on humans (70.2%) than on animals (29.8%) in all of the sites. Sporozoites were only detected in indoor resting An. coluzzii from the Sahel savannah (5.0%) and forest (2.5%) zones. CONCLUSIONS: This study reports high outdoor resting densities of An. gambiae and An. coluzzii with high kdr west mutation frequencies, and the presence of malaria vectors indoors despite the use of long-lasting insecticidal nets and indoor residual spraying. Continuous monitoring of changes in the resting behavior of mosquitoes and the implementation of complementary malaria control interventions that target outdoor resting Anopheles mosquitoes are necessary in Ghana.