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Abstract Background Knockdown resistance (kdr) is one of the primary resistance mechanisms present in anopheline species. Although this mutation is largely spread across the Anopheles gambiae s.l. members, its prevalence in other species is still not well documented. Methods The present study investigated the distribution and allelic frequencies of kdr in An. gambiae s.l., An. pharoensis, and An. ziemanni samples collected in 2022 and 2023 in nine sites spread across five ecogeographical settings in Cameroon. Members of the An. gambiae complex were identified molecularly by polymerase chain reaction (PCR). kdr L1014F and L1014S alleles were screened by PCR and confirmed by sequencing. Results An. gambiae (49.9%), An. coluzzii (36.5%), and An. arabiensis (13%) were identified, and the frequency of the kdr L1014F was high in both An. gambiae and An. coluzzii in all sites. The kdr L1014F allele was detected for the first time in 8 out of 14 An. ziemanni samples examined and in 5 out of 22 An. pharoensis samples examined. The kdr L1014S allele was scarce and found only in the heterozygote “RS” state in An. arabiensis and An. gambiae in Yangah and Santchou. Conclusions The present study sheds light on the rapid expansion of the kdr L1014F allele in malaria vectors in Cameroon and stresses the need for surveillance activities also targeting secondary malaria vectors to improve the control of malaria transmission. Graphical Abstract

Background: The rapid spread of pyrethroid resistance has led to a change in strategy, going from pyrethroid-based nets to PBO + pyrethroid-treated nets. Although these new nets may significantly improve the control of pyrethroid-resistant mosquitoes, their durability in the field remain not yet well documented. This study investigates the durability and washing resistance of Olyset-Plus nets in the city centre and rural areas of Bertoua, Cameroon. In each site, a semi-structured questionnaire was administered to at least 190 households with an Olyset-Plus net. Factors such as net use, physical integrity and bioefficacy were recorded. Bioassays were conducted on the collected nets to assess their bioefficacy and resistance to washing. They were tested against wild Anopheles gambiae sensus lato (s.l.). Unused nets and the Kisumu strain were used as controls. Washing and cone testing of the nets was carried out according to standard WHO protocols. Results: A high rate of net use by children was recorded in the urban area (89.1% (106/119)) compared to the rural area (39.7% (118/297)). The majority of Olyset-Plus nets inspected 82.2% (162/197) in the rural area and 88% (206/234) in the urban centre were in good condition (Hole Index

Abstract Africa and the United States are both large, heterogeneous geographies with a diverse range of ecologies, climates and mosquito species diversity which contribute to disease transmission and nuisance biting. In the United States, mosquito control is nationally, and regionally coordinated and in so much as the Centers for Disease Control (CDC) provides guidance, the Environmental Protection Agency (EPA) provides pesticide registration, and the states provide legal authority and oversight, the implementation is usually decentralized to the state, county, or city level. Mosquito control operations are organized, in most instances, into fully independent mosquito abatement districts, public works departments, local health departments. In some cases, municipalities engage independent private contractors to undertake mosquito control within their jurisdictions. In sub–Saharan Africa (SSA), where most vector-borne disease endemic countries lie, mosquito control is organized centrally at the national level. In this model, the disease control programmes (national malaria control programmes or national malaria elimination programmes (NMCP/NMEP)) are embedded within the central governments’ ministries of health (MoHs) and drive vector control policy development and implementation. Because of the high disease burden and limited resources, the primary endpoint of mosquito control in these settings is reduction of mosquito borne diseases, primarily, malaria. In the United States, however, the endpoint is mosquito control, therefore, significant (or even greater) emphasis is laid on nuisance mosquitoes as much as disease vectors. The authors detail experiences and learnings gathered by the delegation of African vector control professionals that participated in a formal exchange programme initiated by the Pan-African Mosquito Control Association (PAMCA), the University of Notre Dame, and members of the American Mosquito Control Association (AMCA), in the United States between the year 2021 and 2022. The authors highlight the key components of mosquito control operations in the United States and compare them to mosquito control programmes in SSA countries endemic for vector-borne diseases, deriving important lessons that could be useful for vector control in SSA.

Objective: Understanding the epidemiological features of malaria is a key step to monitoring and quantifying the impact of the current control efforts to inform future ones. This study establishes the prevalence and frequency of malaria in a forest–savannah ecozone for 5 consecutive years in Cameroon. Methods: A retrospective study was conducted in 3 health centers of Makenene from 2016 to 2020, a period covering the second long-lasting insecticide net mass distribution campaign. Malaria infectious records were reviewed from laboratory registers. The difference in exposure to malaria was estimated using a regression logistic model. Results: A total of 13525 patients underwent malaria diagnostic tests, with a general malaria prevalence of 65.3%. A greater prevalence of malaria was observed in males (68.39%) compared to females (63.14%). The frequency of consultations in health centers was dominated by females, with a gender ratio (M/F) of 0.66. Annual trends in malaria prevalence slightly varied from 2016 to 2020, exceeding 60%: 65.2% in 2016; 66.7% in 2017, 68.1% in 2018, 63.2% in 2019, and 65.3% in 2020, with a significant seasonal variation (p < 0.0001). The highest malaria prevalence was observed during the short rainy season, no matter the year. Among positive cases, the most represented age groups were 6–15 (p < 0.0001), followed by those under 5, while the age group >25 years was the least represented. Conclusion: Close monitoring and additional intervention measures for malaria control are needed, as are more studies on vector bionomics and transmission patterns.

Human African Trypanosomiasis (HAT) is still endemic in the Republic of Congo. Although the incidence of cases has significantly decreased over years, the disease still persists in some active foci. Factors contributing to the maintenance of the disease such as the existence of an animal reservoir or population knowledge are still not well known. It is in this context that a study focusing on the knowledge and practices of the population with regard to HAT as well as on the prevalence of trypanosomes infecting animals was undertaken in three active HAT foci in the Republic of Congo. The study was performed using field surveys conducted from November 2019 to June 2021. Domestic animal blood was examined by microscopy and PCR to detect the presence of trypanosomes. A structured questionnaire was administered to the population to assess their knowledge and practices concerning HAT in these endemic foci. More than half of the animals examined were found to be infected with trypanosomes (51.22%). The main trypanosome species infecting animals were Trypanosoma congolense savannah (67.2%) and Trypanosoma brucei (s.l.) (32.8%). No trypanosomes infecting humans were detected. Concerning household surveys, more than half of the respondents (52.9%) were fully aware of the mode of transmission and symptoms of the disease. The majority of people preferred to wear clothes covering the whole body and to use locally made soap as repellents to protect themselves from tsetse fly bites. This study suggests frequent circulation of animal trypanosomes in domestic animals and the use of personal measures to protect against tsetse fly bites. Updating information on the HAT animal reservoir and population knowledge alongside regular monitoring of the tsetse fly populations and the use of traps to control tsetse flies are crucial to drive efforts towards the elimination of gHAT in the Republic of Congo.

Abstract Background Malaria remains a major public health concern in Cameroon. Understanding vector distribution and malaria transmission dynamics is of paramount importance for evaluating the performance of control strategies. This study assesses patterns of malaria transmission in four eco-epidemiological settings in Cameroon. Methods Adult mosquitoes were collected using Human Landing Catches (HLC) once every 4 months from August 2019 to November 2021 in Kaélé, Tibati, Santchou and Bertoua. Mosquitoes were sorted by genus and Anopheles gambiae sensu lato (s.l.) species complex were identified using PCR. The presence of Plasmodium falciparum circumsporozoite protein (CSP) was measured by ELISA; the entomological inoculation rates (EIR) was estimated in each locality. Results A total of 23,536 mosquitoes were collected. Anopheles gambiae and/or Anopheles coluzzii were the main malaria vectors in all sites. Anopheles arabiensis was recorded in low frequency in Kaélé and Tibati. Other species collected included Anopheles funestus, Anopheles pharoensis and Anopheles ziemmani. High anopheline biting rates were recorded outdoor in all sites except in Kaélé. Important differences in species biting dynamics were observed between sites. The sporozoite infection rate varied from 0.36 to 4%. The daily EIR was found to vary from 0.07 in Santchou to 0.26 infected bites/man/night (ib/m/n) in Kaélé). Conclusion The study suggests heterogeneous patterns of malaria transmission in different ecoepidemiological settings across the country. The findings stress the need to improve malaria vector control strategies.

Background: The endosymbiotic bacterium Wolbachia infects numerous species of insects and Wolbachia transinfection of Aedes mosquito species is now being used for biocontrol programs as Wolbachia strains can both inhibit arboviruses and invade wild mosquito populations. The discovery of novel, resident Wolbachia strains in mosquito species warrants further investigation as potential candidate strains for biocontrol strategies. Methods: We obtained mosquito specimens from diverse Culicine mosquitoes from Cameroon including ecologically diverse locations in the Central and West Regions. Wolbachia prevalence rates were assessed in addition to the environmentally acquired bacterial species Asaia in major Culicine genera. PCR-based methods were also used with phylogenetic analysis to confirm identities of host mosquito species and Wolbachia strains were classified using multi-locus sequence typing (MLST). Results: We report high Wolbachia prevalence rates for Culicine species, including in a large cohort of Aedes africanus collected from west Cameroon in which 100% of mono-specific pools were infected. Furthermore, co-infections with Asaia bacteria were observed across multiple genera, demonstrating that these two bacteria can co-exist in wild mosquito populations. Wolbachia strain MLST and phylogenetic analysis provided evidence for diverse Wolbachia strains in 13 different mosquito species across seven different genera. Full or partial MLST profiles were generated from resident Wolbachia strains in six Culex species (quinquefasciatus, watti, cinerus, nigripalpus, perexiguus and rima), two Aedes species (africanus and denderensis) and in Mansonia uniformis, Catageiomyia argenteopunctata, Lutzia tigripes, Eretmapodites chrysogaster and Uranotaenia bilineata. Conclusions: Our study provides further evidence that Wolbachia is widespread within wild mosquito populations of diverse Culicine species and provides further candidate strains that could be investigated as future options for Wolbachia-based biocontrol to inhibit arbovirus transmission.

Abstract Background To contribute to the mission of the National Malaria Control Programme (NMCP) and guide future interventions in Cameroon in general, and in Makenene in particular, this study assessed the knowledge, attitudes and practices of the population of Makenene towards the fight against malaria. Methods Using a semi-structured questionnaire, a descriptive cross-sectional household community survey was carried out in randomly selected households in Makenene, a locality situated between forest and savannah ecotypes. Results Out of the 413 households surveyed, all (100%) claimed to have heard of malaria with over 94% (n = 391) associating disease transmission with mosquito bites. The main mosquito control tools used in the area were mosquito nets (92.25%). The majority of participants had good knowledge (55.93%; n = 231), good practices (71.67%, n = 296) but moderate attitudes (47.94%; n = 198) towards malaria control and fight. Good knowledge and practices were recorded mostly in educated persons including public servants and students. Good attitudes were adopted mostly by public servants and students of secondary and higher levels of education. Conclusion In Makenene, the population exhibits good knowledge and practices towards malaria and its control. However, despite high LLINs ownership and use, people still complain about malaria in the area. Control tools should be monitored, repaired or replaced when necessary to support the achievement of the NMCP mission.

Abstract Background The practice of agriculture in urban settings contributes to the rapid expansion of insecticide resistance in malaria vectors. However, there is still not enough information on pesticide usage in most urban settings. The present study aims to assess the evolution of Anopheles gambiae (s.l.) population susceptibility to insecticides and patterns of pesticide usage in agriculture in the city of Yaoundé, Cameroon. Methods WHO susceptibility tests and synergist PBO bioassays were conducted on adult An. gambiae (s.l.) mosquitoes aged 3 to 5 days emerging from larvae collected from the field. Seven insecticides (deltamethrin, permethrin, DDT, bendiocarb, propoxur, fenitrothion and malathion) were evaluated. The presence of target site mutation conferring knockdown (kdr) resistance was investigated using TaqMan assay, and mosquito species were identified using SINE-PCR. Surveys on 81 retailers and 232 farmers were conducted to assess general knowledge and practices regarding agricultural pesticide usage. Results High resistance intensity to pyrethroids was observed with a high frequency of the kdr allele 1014F and low frequency of the kdr 1014S allele. The level of susceptibility of An. gambiae (s.l.) to pyrethroids and carbamates was found to decrease with time (from > 34% in 2017 to

Abstract Background Choosing an appropriate strategy for mosquito vector control in an eco-climatic facies requires knowledge on vector ecology including population dynamics of species and productivity of the breeding sites. The aim of this study was to characterize Culex mosquitoes breeding places and to determine their population dynamics in two localities (Dschang and Santchou) separated by a forest cliff in the West Region of Cameroon. Field surveys were conducted from November 2019 to June 2020 in Dschang and Santchou. Mosquito breeding sites were georeferenced, and for each breeding site, physical parameters were measured and immature stages were collected. The collected immatures were reared in the laboratory until adult stage. Adult mosquitoes were identified using a stereomicroscope and morphological identification keys. Results A total of 44 breeding sites were identified: 24 in Dschang and 20 in Santchou. They were grouped into seven types and were mostly shallow, close to human dwellings, sunny, with organic matters and of anthropogenic nature. A total of 2706 mosquitoes belonging to four genera were identified. Culex genus was the most represented (90.4%) and was made up of Culex pipiens s.l. (61.79%), Culex duttoni (23.17%) and Culex (Culiciomyia) sp. (05.46%). High abundance of Culex species was observed in Santchou (52.71%) compared to Dschang (47.28%), while Dschang (S = 3; H = 0.87; D = 0.54) recorded the greatest diversity compared to Santchou (S = 2; H = 0.23; D = 0.11). Conclusions The proliferation and persistence of Culex mosquitoes independently of the breeding sites and localities might be attributable to the poor environmental management which favor the creation mosquito breeding sites. This study highlights the fact that prompt sanitation measures could be undertaken in these two localities to reduce mosquito abundance and the risk of vector-borne diseases.

Although Culex species are considered to be equally affected by control measures targeting malaria vectors, there is still not enough evidence of the impact of interventions such as larviciding on the distribution of these mosquito species. The present study assessed the impact of a larviciding trial targeting malaria vectors on Culex mosquito species in the city of Yaoundé, Cameroon. A cluster randomized trial comparing 13 treated clusters and 13 untreated clusters was implemented. Data were collected at baseline and during the larviciding intervention, from March 2017 to November 2020. The microbial larvicide VectoMax G was applied once every 2 weeks in the intervention areas. Adult mosquitoes were collected using CDC light traps in both intervention and non-intervention areas and compared between arms. Globally, larviciding intervention was associated with 69% reduction in aquatic habitats with Culex larvae and 36.65% reduction of adult Culex densities in houses. Adult Culex densities were reduced both indoors (35.26%) and outdoors (42.37%). No change in the composition of Culex species was recorded. The study suggests a high impact of larviciding on Culex mosquito species distribution. The impact of the intervention can be improved if typical Culex breeding habitats including pit latrines are targeted.

Abstract The expansion of mosquito-borne diseases such as dengue, yellow fever, and chikungunya in the past 15 years has ignited the need for active surveillance of common and neglected mosquito-borne infectious diseases. The surveillance should be designed to detect diseases and to provide relevant field-based data for developing and implementing effective control measures to prevent outbreaks before significant public health consequences can occur. Mosquitoes are important vectors of human and animal pathogens, and knowledge on their biodiversity and distribution in the Afrotropical region is needed for the development of evidence-based vector control strategies. Following a comprehensive literature search, an inventory of the diversity and distribution of mosquitoes as well as the different mosquito-borne diseases found in Cameroon was made. A total of 290 publications/reports and the mosquito catalogue website were consulted for the review. To date, about 307 species, four subspecies and one putative new species of Culicidae, comprising 60 species and one putative new species of Anopheles, 67 species and two subspecies of Culex, 77 species and one subspecies of Aedes, 31 species and one subspecies of Eretmapodites, two Mansonia, eight Coquillettidia, and 62 species with unknown medical and veterinary importance (Toxorhynchites, Uranotaenia, Mimomyia, Malaya, Hodgesia, Ficalbia, Orthopodomyia, Aedeomyia, and Culiseta and Lutzia) have been collected in Cameroon. Multiple mosquito species implicated in the transmission of pathogens within Anopheles, Culex, Aedes, Eretmapodites, Mansonia, and Coquillettidia have been reported in Cameroon. Furthermore, the presence of 26 human and zoonotic arboviral diseases, one helminthic disease, and two protozoal diseases has been reported. Information on the bionomics, taxonomy, and distribution of mosquito species will be useful for the development of integrated vector management programmes for the surveillance and elimination of mosquito-borne diseases in Cameroon.

Abstract The rapid expansion of insecticide resistance and outdoor malaria transmission are affecting the efficacy of current malaria control measures. In urban settings, where malaria transmission is focal and breeding habitats are few, fixed and findable, the addition of anti-larval control measures could be efficient for malaria vector control. But field evidences for this approach remains scarce. Here we provide findings of a randomized-control larviciding trial conducted in the city of Yaoundé that support the efficacy of this approach. A two arms random control trial design including 26 clusters of 2 to 4 km2 each (13 clusters in the intervention area and 13 in the non-intervention area) was used to assess larviciding efficacy. The microbial larvicide VectoMax combining Bacillus thuringiensis var israelensis (Bti) and Bacillus sphaericus in a single granule was applied every 2 weeks in all standing water collection points. The anopheline density collected using CDC light traps was used as the primary outcome, secondary outcomes included the entomological inoculation rate, breeding habitats with anopheline larvae, and larval density. Baseline entomological data collection was conducted for 17 months from March 2017 to July 2018 and the intervention lasted 26 months from September 2018 to November 2020. The intervention was associated with a reduction of 68% of adult anopheline biting density and of 79% of the entomological inoculation rate (OR 0.21; 95% CI 0.14–0.30, P

Abstract Culex mosquitoes particularly Culex quinquefasciatus are important arboviral and filariasis vectors, however despite this important epidemiological role, there is still a paucity of data on their bionomics. The present study was undertaken to assess the insecticide resistance status of Cx. quinquefasciatus populations from four districts of Yaoundé (Cameroon). All Culex quinquefasciatus populations except one displayed high resistance to bendiocarb and malathion with mortalities ranging from 0 to 89% while high resistance intensity against both permethrin and deltamethrin was recorded. Molecular analyses revealed high frequencies of the ACE-1 G119S mutation (ranging from 0 to 33%) and kdr L1014F allele (ranging from 55 to 74%) in all Cx. quinquefasciatus populations. Significant overexpression was detected for cytochrome P450s genes CYP6AA7 and CYP6Z10, as well as for Esterase A and Esterase B genes. The total cuticular hydrocarbon content, a proxy of cuticular resistance, was significantly increased (compared to the S-lab strain) in one population. The study confirms strong insecticide resistance mediated by different mechanisms in Cx. quinquefasciatus populations from the city of Yaoundé. The expansion of insecticide resistance in Culex populations could affect the effectiveness of current vector control measures and stress the need for the implementation of integrated vector control strategies in urban settings.

Abstract Background Improving house structure is known to limit contact between humans and mosquitoes and reduce malaria transmission risk. In the present study, the influence of house characteristics on mosquito distribution and malaria transmission risk was assessed in the city of Yaoundé. Methods The study was conducted from March 2017 to June 2018 in 32 districts of the city of Yaoundé. Mosquito collections were performed indoor in 10 to 15 houses per district using CDC light traps. A total of 467 houses, selected randomly were used. A pretested questionnaire was submitted to participants of the study to collect information on the household: the number of people per house, education level, type of walls, presence of ceilings and eaves, number of windows, usage of long-lasting insecticidal nets (LLINs), number of bedroom and number of window. Mosquitoes collected were identified morphologically. Anophelines were tested by ELISA to detect infection by Plasmodium parasites. General Estimating Equations adjusting for repeated measures in the same house fitting negative binomial analysis were used to assess the influence of house characteristics on mosquito distribution. Results A total of 168,039 mosquitoes were collected; Culex spp emerged as the predominant species (96.48%), followed by Anopheles gambiae sensu lato (s.l.) (2.49%). Out of the 1033 An. gambiae s.l. identified by PCR, 90.03% were Anopheles coluzzii and the remaining were An. gambiae sensu stricto (s.s.) (9.97%). The high number of people per household, the presence of screens on window and the possession of LLINs were all associated with fewer mosquitoes collected indoors, whilst opened eaves, the high number of windows, the presence of holes in walls and living close to breeding sites were associated with high densities of mosquitoes indoor. Out of 3557 Anophelines tested using ELISA CSP, 80 were found infected by Plasmodium falciparum parasites. The proportion of mosquitoes infected did not vary significantly according to house characteristics. Conclusion The study indicated that several house characteristics such as, the presence of holes on walls, opened eaves, unscreened window and living close to breeding sites, favored mosquito presence in houses. Promoting frequent use of LLINs and house improvement measures, such as the use of screen on windows, closing eaves, cleaning the nearby environment, should be integrated in strategies to improve malaria control in the city of Yaoundé.

Malaria transmission and prevalence is still not well documented across Cameroon particularly in medium-sized cities or localities representing high transit zone. Different risk factors could be associated with persistence malaria transmission such as population movement from high to low transmission settings. A cross-sectional community-based study was carried out to determine malaria prevalence and risk factors in Makenene, a small city in a forest-savannah which is a crossroads between different parts of the country where travellers usually stop-over day and night to rest. Using malaria diagnostic test (mRDTs from SD-BIOLINE) and microscopy (thin and thick blood smears), 406 participants from 237 households were tested for malaria infection. The prevalence of malaria was high irrespective of the detection method: mRDT (41.87%) or microscopy (38.42%). At household level, 46.41% of households had at least one case of malaria with an average of 1.41 infected individuals per household. Parasite density was also high with the majority of infected individuals (64.74%) bearing more than 500 parasites/μl. Only Plasmodium falciparum was found. The chances of being infected with malaria parasites was almost the same for all participants irrespective of the sleeping behavior, bednet usage, house type and environmental factors. The study supports high malaria transmission in the locality and the need for additional studies on vectors bionomics and transmission patterns.

BackgroundUrban malaria is becoming a major public health concern in major cities in Cameroon. To improve malaria vector control, a pilot larviciding trial was conducted to assess its impact on mosquito density and malaria transmission intensity in Yaoundé. The present study investigated perceptions and practices of communities on malaria control during the larviciding trial implemented in Yaoundé.MethodsQuantitative and qualitative data were collected in non-intervention and intervention areas. Quantitative data were collected during three cross-sectional surveys using a structured pre-tested questionnaire while qualitative data were obtained through interviews. A total of 26 in-depth interviews and eight focus group discussions with community members were performed. A binary logistic regression model was used to assess the perception of the community on larviciding impact on some malaria or bed nets use indicators.ResultsPeople living in intervention areas were 2.64 times more likely to know the mode of malaria transmission (95% CI: 1.82-3.84; pConclusionThe larviciding program in the city of Yaoundé did not negatively affected community members' behaviour and practices concerning the use of treated nets. The study indicated the acceptance of larviciding program by the population. This positive environment could favour the implementation of future antilarval control activities in the city of Yaoundé.

Introduction: Human African trypanosomiasis (HAT) is a neglected tropical disease still endemic in the Republic of Congo. Despite the continuous detection of HAT cases in the country, there is still not enough data on trypanosome infections in tsetse flies, trypanosome species and tsetse flies’ species distribution in endemic foci. The present study was intended to fill this gap and improve understanding of trypanosome circulation in three active foci in the centre and south of Congo. Methods: Pyramid traps were set in various places in villages to collect tsetse flies both during the rainy and dry seasons. Once collected, tsetse flies were identified using morphological keys. DNA extracted from flies was processed by PCR for species identification and for detection of trypanosome presence. A second PCR was run for different trypanosome species identification. Results: A total of 1291 tsetse flies were collected. The average apparent density of flies per day was 0.043 in Mpouya, 0.73 in Ngabé and 2.79 in Loudima. Glossina fuscipes quazensis was the predominant tsetse fly collected in Ngabé and Mpouya, while Glossina palpalis palpalis was the only tsetse fly found in Loudima. A total of 224 (17.7%) flies were detected infected by trypanosomes; 100 (7.91%) by Trypanosoma congolense savannah, 22 (1.74%) by Trypanosoma congolense forest, 15 (1.19%) by Trypanosoma vivax, 83 (6.56%) by Trypanosoma brucei (s.l.) and 2 (0.16%) undetermined species. No T Trypanosoma brucei gambiense was found. A total of 57 co-infections between T. brucei (s.l.) and T. congolense savannah or T. brucei (s.l.) and T. congolense forest were found only in G. p. palpalis. Loudima recorded the highest number of infected tsetse flies. Conclusion: The study provided updated information on the distribution of tsetse fly populations as well as on Trypanosoma species circulating in tsetse flies in the different active HAT foci in Congo. These data suggested a high risk of potential transmission of animal trypanosomes in these foci, thus stressing the need for active surveillance in this endemic area.

Abstract Background The rapid and unplanned urbanization of African cities is considered to increase the risk of urban malaria transmission. The present study objective was to assess factors influencing the spatio-temporal distribution of Anopheles gambiae s.l. larvae in the city of Yaoundé, Cameroon. Methods All water bodies were checked once every 2 months for the presence of mosquito larvae from March 2017 to May 2018 in 32 districts of Yaoundé. Physico-chemical characteristics including the size, depth, turbidity, pH, temperature, conductivity, sulfates, organophosphates, hydrogen peroxide (H2O2), conductivity, iron and calcium were recorded and analyzed according to anopheline larvae presence or absence. High resolution satellite images from landsat sentinel Enhanced Thematic Mapper were used for spatial mapping of both field and environmental variables. Bivariate and multivariate logistic regression models were used to identify variables closely associated with anopheline larvae distribution. Results A total of 18 696 aquatic habitats were checked and only 2942 sites (15.7%) contained anopheline larvae. A high number of sites with anopheline larvae (≥ 69%) presented late instar larvae (L3, L4 and pupae). Anopheline mosquito larvae were sampled from a variety of breeding sites including puddles (51.6%), tire prints (12.9%), wells (11.7%) and drains (11.3%). Bivariate logistic regression analyses associated anopheline larvae presence with the absence of predators, absence of algae, absence of vegetation and depth of less than 1 m. Conductivity, turbidity, organophosphates, H2O2 and temperature were significantly high in breeding sites with anopheline larvae than in breeding sites without these larvae (P

Abstract Malaria still has a devastating impact on public health and welfare in Cameroon. Despite the increasing number of studies conducted on disease prevalence, transmission patterns or treatment, there are to date, not enough studies summarising findings from previous works in order to identify gaps in knowledge and areas of interest where further evidence is needed to drive malaria elimination efforts. The present study seeks to address these gaps by providing a review of studies conducted so far on malaria in Cameroon since the 1940s to date. Over 250 scientific publications were consulted for this purpose. Although there has been increased scale-up of vector control interventions which significantly reduced the morbidity and mortality to malaria across the country from a prevalence of 41% of the population reporting at least one malaria case episode in 2000 to a prevalence of 24% in 2017, the situation is not yet under control. There is a high variability in disease endemicity between epidemiological settings with prevalence of Plasmodium parasitaemia varying from 7 to 85% in children aged 6 months to 15 years after long-lasting insecticidal nets (LLINs) scale-up. Four species of Plasmodium have been recorded across the country: Plasmodium falciparum, P. malariae, P. ovale and P. vivax. Several primate-infecting Plasmodium spp. are also circulating in Cameroon. A decline of artemisinin-based combinations therapeutic efficacy from 97% in 2006 to 90% in 2016 have been reported. Several mutations in the P. falciparum chloroquine resistance (Pfcrt) and P. falciparum multidrug resistance 1 (Pfmdr1) genes conferring resistance to either 4-amino-quinoleine, mefloquine, halofanthrine and quinine have been documented. Mutations in the Pfdhfr and Pfdhps genes involved in sulfadoxine-pyrimethamine are also on the rise. No mutation associated with artemisinin resistance has been recorded. Sixteen anopheline species contribute to malaria parasite transmission with six recognized as major vectors: An. gambiae, An. coluzzii, An. arabiensis, An. funestus, An. nili and An. moucheti. Studies conducted so far, indicated rapid expansion of DDT, pyrethroid and carbamate resistance in An. gambiae, An. coluzzii, An. arabiensis and An. funestus threatening the performance of LLINs. This review highlights the complex situation of malaria in Cameroon and the need to urgently implement and reinforce integrated control strategies in different epidemiological settings, as part of the substantial efforts to consolidate gains and advance towards malaria elimination in the country.

Abstract Background Malaria prevention in Cameroon mainly relies on the use of ITNs. Although several free distribution campaigns of treated nets have been conducted across the country, bed net usage remains very low. A household survey was conducted to assess knowledge of the population and practices affecting treated net usage in the city of Yaoundé. Methods A community-based descriptive cross-sectional survey was conducted in January 2017 in 32 districts of the city of Yaoundé. Parents (household head, spouse or an elder representative) who consented to the study, were interviewed using a structured pre-tested questionnaire. Interviews were conducted in French or English. A questionnaire consisting of 22 questions was administered to know (i) people’s knowledge and attitude on preventive measures; and, (ii) attitudes concerning the treatment of malaria and estimated amount spent for malaria prevention and treatment. Results A total of 1643 household heads were interviewed. Over 94% of people interviewed associated malaria transmission to mosquito bites. The main methods used against mosquito bites were: treated bed nets (94%; n = 1526) and insecticide spray or coils (32.2%; n = 523). The majority of people interviewed reported using bed nets mainly to prevent from mosquito bites (84.4%, n = 1257), rather than for malaria prevention (47.3%). Knowledge and attitude analysis revealed that people with university or secondary level of education have better knowledge of malaria, prevention and treatment measures compared to those with the primary level (OR = 7.03; P

Dengue and chikungunya are now widely distributed in Cameroon, but there is still not enough information on their prevalence in different epidemiological settings. This study was undertaken to assess the prevalence of dengue and chikungunya in both urban and rural settings in Cameroon using three diagnostic tools. From December 2019 to September 2021, willing febrile (temperature >38 °C) outpatients visiting four healthcare facilities in the cities of Yaoundé and Dizangué were screened for dengue, and chikungunya. Clinical features of patient were recorded in a form, and their blood samples were analysed using real-time reverse transcriptase-polymerase chain reaction (rtRT-PCR), rapid diagnostic tests (RDTs) and enzyme-linked immuno-sorbent assays (ELISA). Odds ratios were used to determine the level of association between socio-demographic factors, clinical features, and infection status. The Kappa coefficient permitted to assess the level of agreement between RDTs and ELISA. Overall, 301 febrile patients were recruited in the study: 198 in Yaoundé and 103 in Dizangué. The prevalence of infection varied with the diagnostic tool used. For dengue diagnostics, 110 patients were positive to rtRT-PCR: 90 (45.45%) in Yaoundé, and 20 (19.42%) in Dizangué. The prevalence of dengue IgM using ELISA varied from 22.3% in Dizangué to 30.8% in Yaoundé. Dengue IgM rate using RDTs was 7.6% in Yaoundé and 3.9% in Dizangué. For chikungunya, one (0.5%) patient (Yaoundé, suburb) was positive to rtRT-PCR. The prevalence of chikungunya IgM according to ELISA varied from 18.4% in Dizangué to 21.7% in Yaoundé, while it was 4.5% in Yaoundé and 12.6% in Dizangué with RDTs. Only abdominal and retro-orbital pains were significantly associated with acute dengue infection. All four dengue serotypes were recorded, with a predominance of DENV-3 (35.45%) and DENV-4 (25.45%). Rapid Diagnostic Tests for either chikungunya or dengue displayed very poor sensitivity. This study further confirms the high endemicity of both dengue and chikungunya in Yaoundé and Dizangué. These data stress the need for active surveillance and the implementation of vector control measures to prevent the occurrence of outbreaks across the country.

Rapid emergence and spread of pyrethroid resistance in Anopheles gambiae populations is among the main factors affecting malaria vector control in Cameroon, but there is still not enough data on the exact pyrethroid resistance status across Cameroon. The present study assessed pyrethroid resistance profile in different eco-epidemiological settings in Cameroon. Susceptibility bioassay tests were performed with F0 An. gambiae females aged three to five days. Mosquito susceptibility to both permethrin and deltamethrin was assessed. Species of the An. gambiae s.l. complex were identified using molecular diagnostic tools. Target site mutations conferring resistance were detected using Taqman assays. Quantitative reverse transcription-real-time PCR (qRT-PCR) 3-plex TaqMan® assays were used for the quantification of detoxification genes implicated in pyrethroid resistance. An. gambiae, An. coluzzii and An. arabiensis were identified in the different settings. An. gambiae was dominant in Santchou, Kékem, Bélabo, Bertoua and Njombé, while An. coluzzii was abundant in Tibati and Kaélé. High frequencies of the kdr L1014F allele ranging from 43% to 100% were recorded in almost all sites. The L1014S kdr allele was detected at low frequency (4.10–10%) only in mosquito populations from Njombé and Tibati. The N1575Y mutation was recorded in Kaélé, Santchou, Tibati and Bertoua with a frequency varying from 2.10% to 11.70%. Six Cytochrome P450 genes (Cyp6p3, Cyp6m2, Cyp9k1, Cyp6p4, Cyp6z1, and Cyp4g16) were found to be overexpressed in at least one population. Analysis of cuticular hydrocarbon lipids indicated a significant increase in CHC content in mosquito populations from Kaélé and Njombé compared to Kékem, Bélabo and Bertoua populations. The study indicated high pyrethroid resistance across different ecological settings in Cameroon with different profile of resistance across the country. The present situation calls for further actions in order to mitigate the impact of insecticide resistance on vector control measures.

Arbovirus diseases represent a significant public health problem in Cameroon and vector surveillance is a key component of prevention strategies. However, there is still not enough evidence of the efficacy of different sampling methods used to monitor Aedes mosquito population dynamic in different epidemiological settings. The present study provides data on the evaluation of ovitraps and different adult sampling methods in the city of Yaoundé and its close vicinity. Entomological surveys were carried out from February 2020 to March 2021 in two urban (Obili, Mvan), two peri-urban (Simbock, Ahala), and two rural (Lendom, Elig-essomballa) sites in the city of Yaoundé. The efficacy of three sampling methods, namely ovitraps, Biogent Sentinel trap, and sweep nets, was evaluated. Different ovitrap indices were used to assess the infestation levels across study sites; a general linear model was used to determine if there are statistical differences between positive ovitraps across ecological zones. A total of 16,264 Aedes mosquitoes were collected during entomological surveys. Ovitraps provided the highest mosquito abundance (15,323; 91.14%) and the highest species diversity. Of the five Aedes species collected, Aedes albopictus (59.74%) was the most commonly recorded in both urban and rural settings. Different Aedes species were collected in the same ovitrap. The ovitrap positivity index was high in all sites and varied from 58.3% in Obili in the urban area to 86.08% in Lendom in the rural area. The egg density index varied from 6.42 in Mvan (urban site) to 13.70 in Lendom (rural area). Adult sampling methods recorded mostly Aedes albopictus. The present study supports high infestation of Aedes species in the city of Yaoundé. Ovitraps were highly efficient in detecting Aedes distribution across study sites. The situation calls for regular surveillance and control of Aedes population to prevent sudden occurrence of outbreaks.

The spread of pyrethroid resistance in malaria vectors is a major threat affecting the performance of current control measures. However, there is still not enough information on the resistance profile of mosquitoes to carbamates and organophosphates which could be used as alternatives. The present study assessed the resistance profile of Anopheles gambiae s.l. to bendiocarb and malathion, at the phenotypic and molecular levels, in different eco-epidemiological settings in Cameroon. Anopheles gambiae s.l. mosquitoes were collected from four eco-epidemiological settings across the country and their susceptibility level to bendiocarb and malathion was determined using WHO tubes bioassays. The ace-1 target site G119S mutation was screened by PCR. Reverse Transcription quantitative PCR 3-plex TaqMan assays were used to quantify the level of expression of eight genes associated with metabolic resistance. Resistance to malathion and/or bendiocarb was recorded in all study sites except in mosquitoes collected in Kaélé and Njombé. The Ace-1 (G119S) mutation was detected in high frequencies (>40%) in Kékem and Santchou. Both An. gambiae and An. coluzzii were detected carrying this mutation. The cytochrome P450s gene Cyp6p3 associated with carbamate resistance and the glutathione S-transferase gene Gste2 associated with organophosphate resistance were found to be overexpressed. Genes associated with pyrethroid (Cyp6m2, Cyp9k1, Cyp6p3) and organochlorine (Gste2, Cyp6z1, Cyp6m2) and cuticle resistance (Cyp4g16) were also overexpressed. The rapid spread of resistance to organophosphates and carbamates could seriously compromise future control strategies based on IRS. It is therefore becoming important to assess the magnitude of bendiocarb and malathion resistance countrywide.

Background: Pyrethroid resistance is rapidly expanding in An. gambiae s.l. populations across Sub-Saharan Africa. Yet there is still not enough information on the fitness cost of insecticide resistance . In the present study, the fitness cost of insecticide resistance on Anopheles coluzzii population from the city of Yaoundé was investigated. Methods: A resistant An. coluzzii colony was established from field collected mosquitoes resistant to both DDT and pyrethroid and selected for 12 generations with deltamethrin 0.05%. The Ngousso laboratory susceptible strain was used as control. A total of 100 females of each strain were blood fed and allowed for individual eggs laying, and then different life traits parameters such as fecundity, fertility, larval development time, emergence rate and longevity were measured. The TaqMan assay was used to screen for the presence of the L1014F and L1014S kdr mutations. Results: Field collected mosquitoes from the F0 generation had a mortality rate of 2.05% for DDT, 34.16% for permethrin and 50.23% for deltamethrin. The mortality rate of the F12 generation was 30.48% for deltamethrin, 1.25% for permethrin and 0% for DDT. The number of eggs laid per female was lower in the resistant colony compared to the susceptible (p

Abstract Background Increased use of long-lasting insecticidal nets (LLINs) over the last decade has considerably improved the control of malaria in sub-Saharan Africa. However, there is still a paucity of data on the influence of LLIN use and other factors on mosquito bionomics in different epidemiological foci. The objective of this study was to provide updated data on the evolution of vector bionomics and malaria transmission patterns in the equatorial forest region of Cameroon over the period 2000–2017, during which LLIN coverage has increased substantially. Methods The study was conducted in Olama and Nyabessan, two villages situated in the equatorial forest region. Mosquito collections from 2016–2017 were compared to those of 2000–2001. Mosquitoes were sampled using both human landing catches and indoor sprays, and were identified using morphological taxonomic keys. Specimens belonging to the An. gambiae complex were further identified using molecular tools. Insecticide resistance bioassays were undertaken on An. gambiae to assess the susceptibility levels to both permethrin and deltamethrin. Mosquitoes were screened for Plasmodium falciparum infection and blood-feeding preference using the ELISA technique. Parasitological surveys in the population were conducted to determine the prevalence of Plasmodium infection using rapid diagnostic tests. Results A change in the species composition of sampled mosquitoes was recorded between the 2000–2001 collections and those of 2016–2017. A drop in the density of the local primary vectors An. nili and An. moucheti in the forest region was recorded, whereas there was an increase in the density of An. gambiae (s.l.), An. marshallii, An. ziemannii and An. paludis. A change in the biting behaviour from indoor to outdoor was recorded in Olama. Very few indoor resting mosquitoes were collected. A change in the night biting cycle was recorded with mosquitoes displaying a shift from night biting to late evening/early in the night. Several mosquitoes were found positive for Plasmodium infection, thus sustaining continuous transmission of malaria in both sites. Reduction of malaria transmission in Nyabessan was lower than that seen in Olama and associated with deforestation and the construction of a dam that may have enabled a more efficient vector, An. gambiae (s.l.), to invade the area. A high level of resistance to pyrethroids (permethrin and deltamethrin) was detected for An. gambiae in both sites. High parasite prevalence was recorded in both sites, with children of 0–16 years being the most affected. In both Olama and Nyabessan, bed net usage appeared to correlate to protection against malaria infection. Conclusions The study shows important changes in the bionomics of vector populations and malaria transmission patterns in the equatorial forest region. The changes call for more concerted efforts to address challenges such as insecticide resistance, environmental modifications or behavioural changes affecting the performance of current control measures.

Abstract Background Following the recent discovery of the role of Anopheles rufipes Gough, 1910 in human malaria transmission in the northern savannah of Cameroon, we report here additional information on its feeding and resting habits and its susceptibility to the pyrethroid insecticide deltamethrin. Methods From 2011 to 2015, mosquito samples were collected in 38 locations across Garoua, Mayo Oulo and Pitoa health districts in North Cameroon. Adult anophelines collected using outdoor clay pots, window exit traps and indoor spray catches were checked for feeding status, blood meal origin and Plasmodium circumsporozoite protein. The susceptibility of field-collected An. rufipes to deltamethrin was assessed using WHO standard procedures. Results Of 9327 adult Anopheles collected in the 38 study sites, An. rufipes (6.5%) was overall the fifth most abundant malaria vector species following An. arabiensis (52.4%), An. funestus (s.l.) (20.8%), An. coluzzii (12.6%) and An. gambiae (6.8%). This species was found outdoors (51.2%) or entering houses (48.8%) in 35 suburban and rural locations, together with main vector species. Apart from human blood with index of 37%, An. rufipes also fed on animals including cows (52%), sheep (49%), pigs (16%), chickens (2%) and horses (1%). The overall parasite infection rate of this species was 0.4% based on the detection of P. falciparum circumsporozoite proteins in two of 517 specimens tested. Among the 21 An. rufipes populations assessed for deltamethrin susceptibility, seven populations were classified as “susceptible” (mortality ≥ 98%) , ten as “probable resistant” with a mortality range of 90–97% and four as “resistant” with a mortality range of 80–89%. Conclusions This study revealed changeable resting and feeding behaviour of An. rufipes, as well as further evidence on its ability to carry human malaria parasites in North Cameroon. Besides, this species is developing physiological resistance to deltamethrin insecticide which is used in treated nets and agriculture throughout the country, and should be regarded as one of potential targets for the control of residual malaria parasite transmission in Africa.

Abstract During the last decade, scale-up of vector control tools such as long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) contributed to the reduction of malaria morbidity and mortality across the continent. Because these first line interventions are now affected by many challenges such as insecticide resistance, change in vector feeding and biting behaviour, outdoor malaria transmission and adaptation of mosquito to polluted environments, the World Health Organization recommends the use of integrated control approaches to improve, control and elimination of malaria. Larviciding is one of these approaches which, if well implemented, could help control malaria in areas where this intervention is suitable. Unfortunately, important knowledge gaps remain in its successful application. The present review summarises key parameters that should be considered when implementing larviciding efficacy or effectiveness trials.

Wolbachia spp., known to be maternally inherited intracellular bacteria, are widespread among arthropods, including mosquitoes. Our study assessed the presence and prevalence of Wolbachia infection in wild mosquitoes collected in Cameroon, using the combination of 23s rRNA Anaplasmatacea and 16s rRNA Wolbachia genes. Mosquitoes that were positive for Wolbachia were sequenced for subsequent phylogenetic analysis. Out of a total of 1740 individual mosquitoes belonging to 22 species and five genera screened, 33 mosquitoes (1.87%) belonging to eight species (namely, Aedes albopictus, A. contigus, Culex quinquefasciatus, C. perfuscus, C. wigglesworthi, C. duttoni, Anopheles paludis and Coquillettidia sp.) were found to be positive for Wolbachia infections. Wolbachia spp. were absent in A. gambiae and A. aegypti, the main vectors of malaria and dengue, respectively. Phylogenetic analysis of the 16S RNA sequences showed they belong mainly to two distinct subgroups (A and B). This study reports the presence of Wolbachia in about eight species of mosquitoes in Cameroon and suggests that future characterisation of the strains is needed.

Introduction: The surveillance of mosquito vectors is important for the control of arboviruses diseases worldwide. Detailed information on the bionomics and distribution of their main vectors, Aedes aegypti and Aedes albopictus, is essential for assessing disease transmission risk and for better planning of control interventions. Methods: Entomological surveys were carried out from November 2019 to November 2020 in six localities of Yaoundé city following a transect from urban to rural settings: two urban (Obili, Mvan), two peri-urban (Simbock, Ahala) and two rural areas (Lendom, Elig-essomballa)—during rainy and dry seasons. All water containers were inspected. Aedes mosquito abundance, species distribution and seasonal distribution patterns were compared using generalized linear models. Stegomyia indexes were estimated to determine the risk of arbovirus transmission. Results: A total of 6332 mosquitoes larvae were collected (2342 in urban areas, 1694 in peri-urban areas and 2296 in rural sites). Aedes species recorded included Ae. albopictus, Ae. aegytpi, Ae. simpsoni and Aedes spp. High mosquito abundance was registered in the rainy season (4706) compared to the dry season (1626) (p < 0.0001). Ae. albopictus was the most abundant Aedes species in urban (96.89%) and peri-urban (95.09%) sites whereas Ae. aegypti was more prevalent in rural sites (68.56%) (p < 0.0001). Both species were found together in 71 larval habitats. Ae. albopictus was mostly found in discarded tires (42.51%), whereas Ae. aegypti was more prevalent in plastic containers used for storing water (65.87%). The majority of Aedes mosquitoes’ breeding places were situated close to human dwellings (0–10 m). Conclusion: Uncontrolled urbanization seems to greatly favour the presence of Aedes mosquito species around human dwellings in Yaoundé. Controlling Aedes mosquito distribution is becoming urgent to reduce the risk of arbovirus outbreaks in the city of Yaoundé.

Abstract Background Malaria remains a major public health threat in Cameroon and disease prevention is facing strong challenges due to the rapid expansion of insecticide resistance in vector populations. The present review presents an overview of published data on insecticide resistance in the main malaria vectors in Cameroon to assist in the elaboration of future and sustainable resistance management strategies. Methods A systematic search on mosquito susceptibility to insecticides and insecticide resistance in malaria vectors in Cameroon was conducted using online bibliographic databases including PubMed, Google and Google Scholar. From each peer-reviewed paper, information on the year of the study, mosquito species, susceptibility levels, location, insecticides, data source and resistance mechanisms were extracted and inserted in a Microsoft Excel datasheet. The data collected were then analysed for assessing insecticide resistance evolution. Results Thirty-three scientific publications were selected for the analysis. The rapid evolution of insecticide resistance across the country was reported from 2000 onward. Insecticide resistance was highly prevalent in both An. gambiae (s.l.) and An. funestus. DDT, permethrin, deltamethrin and bendiocarb appeared as the most affected compounds by resistance. From 2000 to 2017 a steady increase in the prevalence of kdr allele frequency was noted in almost all sites in An. gambiae (s.l.), with the L1014F kdr allele being the most prevalent. Several detoxification genes (particularly P450 monooxygenase) were associated with DDT, pyrethroids and bendiocarb resistance. In An. funestus, resistance to DDT and pyrethroids was mainly attributed to the 119F-GSTe2 metabolic resistance marker and over-expression of P450 genes whereas the 296S-RDL mutation was detected in dieldrin-resistant An. funestus. Conclusions The review provides an update of insecticide resistance status in malaria vector populations in Cameroon and stresses the need for further actions to reinforce malaria control strategies in the coming years.

BACKGROUND:Culex species are widespread across Cameroon and responsible for high burden of nuisance in most urban settings. However, despite their high nuisance, they remain less studied compared to anophelines. The present study aimed to assess Culex species distribution, susceptibility to insecticide, bionomics and role in Lymphatic Filariasis (LF) transmission in the city of Yaoundé. METHODS:Mosquito collections were conducted from March to December 2017 using Centre for Disease Control light traps (CDC-LT), human landing catches (HLC) and larval collections. Mosquitoes were identified using morphological identification keys. Mosquitoes from the Culex pipiens complex were further identified using Polymerase Chain Reaction (PCR) to assess the presence of sibling species. Bioassays were conducted with 2-5 day-old unfed females to assess mosquito susceptibility to DDT, permethrin, deltamethrin and bendiocarb following WHO guidelines. Dead, control and surviving mosquitoes from bioassays were screened by PCR to detect the presence of knockdown resistance (kdr) alleles. Pools of mosquitoes were examined by PCR to detect the presence of Wuchereria bancrofti. RESULTS:A total of 197,956 mosquitoes belonging to thirteen species were collected. The density of mosquito collected varied according to the collection methods, districts and seasons. Culex quinquefasciatus emerged as the most abundant and the only species of the Culex pipiens complex in Yaoundé. Culex species were found breeding in different types of breeding sites including polluted and unpolluted sites. All Culex species including Cx antennatus, Cx duttoni, Cx perfuscus and Cx tigripes were found to be highly resistant to permethrin, deltamethrin and DDT. Culex quinquefasciatus was also found to be resistant to bendiocarb. A high frequency of the West Africa kdr allele was recorded in resistant Cx. quinquefasciatus. Out of the 247 pooled samples of 25 Culex spp. examined for the presence of Wuchereria bancrofti, none was found infected. CONCLUSION:The study confirms the high adaptation of Culex species particularly Culex quinquefasciatus to the urban environment and no implication of this species in the transmission of LF in Yaoundé Cameroon. Culex species predominance in urban settings highlight potential transmission risk of West Nile and rift valley fever in Yaoundé.

To identify potential sylvatic, urban and bridge-vectors that can be involved in current or future virus spillover from wild to more urbanised areas, entomological field surveys were conducted in rural, peri-urban and urban areas spanning the rainy and dry seasons in western Cameroon. A total of 2650 mosquitoes belonging to 37 species and eight genera were collected. Mosquito species richness was significantly influenced by the specific combination of the habitat type and the season. The highest species richness was found in the peri-urban area (S = 30, Chao1 = 121 ± 50.63, ACE = 51.97 ± 3.88) during the dry season (S = 28, Chao1 = 64 ± 25.7, ACE = 38.33 ± 3.1). Aedes (Ae.) africanus and Culex (Cx.) moucheti were only found in the rural and peri-urban areas, while Cx. pipiens s.l. and Ae. aegypti were only found in the urban area. Cx. (Culiciomyia) spp., Cx. duttoni and Ae. albopictus were caught in the three habitat types. Importantly, approximately 52% of the mosquito species collected in this study have been implicated in the transmission of diverse arboviruses. This entomological survey provides a catalogue of the different mosquito species that may be involved in the transmission of arboviruses. Further investigations are needed to study the vectorial capacity of each mosquito species in arbovirus transmission.

Background: Malaria remains a major public health problem in Cameroon; however, despite reports on the adaptation of anopheline species to urban habitats, there is still not enough information on malaria transmission pattern in urban settings. In the frame of a larval control trial in the city of Yaoundé, we conducted baseline surveys to assess malaria transmission dynamics in this city. Methods: Adult mosquitoes were collected indoors and outdoors using CDC light traps and human landing catches from March 2017 to March 2018 in 30 districts of Yaoundé, Cameroon. Mosquitoes were sorted by genus and identified to the species level using PCR. The TaqMan method and ELISA were used to determine mosquito infection status to Plasmodium. Bioassays were conducted to assess female Anopheles gambiae susceptibility to insecticides. Results: A total of 218,991 mosquitoes were collected. The main malaria vectors were An. gambiae s.l. (n=6154) and An. funestus s.l. (n=229). Of the 1476 An. gambiae s.l. processed by PCR, 92.19% were An. coluzzii and 7.81% An. gambiae. An. funestus s.l. was composed of 93.01% (173/186) An. funestus and 4.84% (13/186) An. leesoni. The average biting rate of anopheline was significantly high outdoor than indoor (P=0.013). Seasonal variation in mosquito abundance and biting rate was recorded. The infection rate by Plasmodium falciparum was 2.13% (104/4893 mosquitoes processed). The annual entomological inoculation rate was found to vary from 0 to 92 infective bites/man/year (ib/m/y). Malaria transmission risk was high outdoor (66.65 ib/m/y) compared to indoor (31.14 ib/m/y). An. gambiae s.l. was found highly resistant to DDT, permethrin and deltamethrin. High prevalence of the West Africa kdr allele 1014F was recorded and this was not found to influence An. gambiae s.l. infection status. Conclusion: The study suggests high malaria transmission occurring in the city of Yaoundé and call for immediate actions to improve control strategies.

INTRODUCTION: A routine diagnosis of dengue virus (DENV) infection is not usually conducted in hospitals. Because symptoms overlap, many potential febrile illnesses due to DENV may be confused for malaria, typhoid or paratyphoid (enteric) fever. The absence of data on DENV exposure rates among children attending health facilities could undermine management of this disease. This study aimed to investigate the seroprevalence of dengue virus infection in children presenting febrile illness in some public health facilities in Cameroon. METHODS: a cross-sectional study was performed in children ≤ 15 years attending seven urban and three semi-urban public hospitals of Cameroon. From each volunteer, 2ml of whole blood was collected and tested for dengue virus IgM, malaria (Pf/Pan antigens) and enteric fever (Typhoid IgM) using rapid diagnostic tests (RDT); in order to allow the healthcare workers to quickly put the positive cases under appropriate treatment. Positive cases of dengue virus infection were confirmed by indirect ELISA. Data analysis were performed using the statistical package for social sciences software, version 22.1. RESULTS: a total of 961 children were enrolled in the study and 492 (51.2%) were infected with at least one of the three pathogens. Overall, DENV IgM seroprevalence among febrile children was 14.4% (138/961). About 390 (40.6%) and 22 (2.3%) had malaria (Pf/Pan Ag) and enteric fever (Typhoid IgM) respectively. Co-infection with dengue virus was found in 51 (5.3%) participants. The dengue virus IgM seroprevalence was higher in Bankim (19.3%), Ntui (18.3%) and Douala (18.2%). CONCLUSION: dengue virus infection seroprevalence appears to be low in children presenting with febrile illness in the studied health centres in Cameroon but call for more attention and research to further characterise the circulating strains of the dengue virus.

Following publication of the original article [1], the authors flagged an error concerning Table 3.

BACKGROUND:Despite the recent progress in establishing the patterns of insecticide resistance in the major malaria vector Anopheles funestus, Central African populations of this species remain largely uncharacterised. To bridge this important gap and facilitate the implementation of suitable control strategies against this vector, we characterised the resistance patterns of An. funestus population from northern Cameroon. METHODS AND FINDINGS:Collection of indoor-resting female mosquitoes in Gounougou (northern Cameroon) in 2012 and 2015 revealed a predominance of An. funestus during dry season. WHO bioassays performed using F1 An. funestus revealed that the population was multiple resistant to several insecticide classes including pyrethroids (permethrin, deltamethrin, lambda-cyhalothrin and etofenprox), carbamates (bendiocarb) and organochlorines (DDT and dieldrin). However, a full susceptibility was observed against the organophosphate malathion. Bioassays performed with 2015 collection revealed that resistance against pyrethroids and DDT is increasing. PBO synergist assays revealed a significant recovery of susceptibility for all pyrethroids but less for DDT. Analysis of the polymorphism of a portion of the voltage-gated sodium channel gene (VGSC) revealed the absence of the L1014F/S kdr mutation but identified 3 novel amino acid changes I877L, V881L and A1007S. However, no association was established between VGSC polymorphism and pyrethroid/DDT resistance. The DDT resistant 119F-GSTe2 allele (52%) and the dieldrin resistant 296S-RDL allele (45%) were detected in Gounougou. Temporal analysis between 2006, 2012 and 2015 collections revealed that the 119F-GSTe2 allele was relatively stable whereas a significant decrease is observed for 296S-RDL allele. CONCLUSION:This multiple resistance coupled with the temporal increased in resistance intensity highlights the need to take urgent measures to prolong the efficacy of current insecticide-based interventions against An. funestus in this African region.

The major vectors of malaria in sub-Saharan Africa belong to subgenus Cellia. Yet, phylogenetic relationships and temporal diversification among African mosquito species have not been unambiguously determined. Knowledge about vector evolutionary history is crucial for correct interpretation of genetic changes identified through comparative genomics analyses. In this study, we estimated a molecular phylogeny using 49 gene sequences for the African malaria vectors An. gambiae, An. funestus, An. nili, the Asian malaria mosquito An. stephensi, and the outgroup species Culex quinquefasciatus and Aedes aegypti. To infer the phylogeny, we identified orthologous sequences uniformly distributed approximately every 5 Mb in the five chromosomal arms. The sequences were aligned and the phylogenetic trees were inferred using maximum likelihood and neighbor-joining methods. Bayesian molecular dating using a relaxed log normal model was used to infer divergence times. Trees from individual genes agreed with each other, placing An. nili as a basal clade that diversified from the studied malaria mosquito species 47.6 million years ago (mya). Other African malaria vectors originated more recently, and independently acquired traits related to vectorial capacity. The lineage leading to An. gambiae diverged 30.4 mya, while the African vector An. funestus and the Asian vector An. stephensi were the most closely related sister taxa that split 20.8 mya. These results were supported by consistently high bootstrap values in concatenated phylogenetic trees generated individually for each chromosomal arm. Genome-wide multigene phylogenetic analysis is a useful approach for discerning historic relationships among malaria vectors, providing a framework for the correct interpretation of genomic changes across species, and comprehending the evolutionary origins of this ubiquitous and deadly insect-borne disease.

BackgroundIn the city of Yaoundé in Cameroon malaria is predominately transmitted by the M and S molecular forms of Anopheles gambiae and both are resistant to the pyrethroid insecticides and DDT. Mutations in the target site of these insecticides, present at a high frequency in malaria vectors in this city, contribute to this resistance profile. To identify additional resistance mechanisms, the expression profile of multiple DDT-resistant field populations of M and S molecular forms was compared to laboratory-susceptible populations.Methodology/principal findingsThe prevalence of DDT resistance was highest in the S form population originating from the cultivated site of Nkolondom (mortality after WHO bioassay = 4%). A high prevalence of DDT resistance was also found in two urban M form populations, Messa from a pristine unpolluted environment (DDT mortality = 54%), and Gare, where the breeding sites are heavily polluted with organic matter (DDT mortality = 38%). Microarray analysis showed that several transcripts coding for detoxification enzymes (P450s, GSTs and UDPGTs) and ABC transporters were upregulated in the three populations. Despite the presence of multiple detoxification genes over expressed in the DDT-resistant subset of these field populations, only three were commonly over expressed in resistant populations from all three environments. Two of these genes, CYP6M2 and GSTD1-6, encode enzymes that have been previously shown to metabolize DDT.Conclusion/significanceAnalogous to target site resistance, genes involved in metabolic resistance to DDT are also shared between the M and S forms of An gambiae. Alternative explanations for this occurrence are explored.

The Anopheles nili group of mosquitoes includes important vectors of human malaria in equatorial forest and humid savannah regions of sub-Saharan Africa. However, it remains largely understudied, and data on its populations' bionomics and genetic structure are crucially lacking. Here, we used a combination of nuclear (i.e. microsatellite and ribosomal DNA) and mitochondrial DNA markers to explore and compare the level of genetic polymorphism and divergence among populations and species of the group in the savannah and forested areas of Cameroon, Central Africa.All the markers provided support for the current classification within the An. nili group. However, they revealed high genetic heterogeneity within An. nili s.s. in deep equatorial forest environment. Nuclear markers showed the species to be composed of five highly divergent genetic lineages that differed by 1.8 to 12.9% of their Internal Transcribed Spacer 2 (ITS2) sequences, implying approximate divergence time of 0.82 to 5.86 million years. However, mitochondrial data only detected three major subdivisions, suggesting different evolutionary histories of the markers.This study enlightened additional cryptic genetic diversity within An. nili s.s. in the deep equatorial forest environment of South Cameroon, reflecting a complex demographic history for this major vector of malaria in this environment. These preliminary results should be complemented by further studies which will shed light on the distribution, epidemiological importance and evolutionary history of this species group in the African rainforest, providing opportunities for in-depth comparative studies of local adaptation and speciation in major African malaria vectors.

BackgroundAnthropogenic habitat disturbance is a prime cause in the current trend of the Earth's reduction in biodiversity. Here we show that the human footprint on the Central African rainforest, which is resulting in deforestation and growth of densely populated urban agglomerates, is associated to ecological divergence and cryptic speciation leading to adaptive radiation within the major malaria mosquito Anopheles gambiae.Methodology/principal findingsIn southern Cameroon, the frequency of two molecular forms--M and S--among which reproductive isolation is strong but still incomplete, was correlated to an index of urbanisation extracted from remotely sensed data, expressed as the proportion of built-up surface in each sampling unit. The two forms markedly segregated along an urbanisation gradient forming a bimodal cline of ∼6-km width: the S form was exclusive to the rural habitat, whereas only the M form was present in the core of densely urbanised settings, co-occurring at times in the same polluted larval habitats of the southern house mosquito Culex quinquefasciatus--a species association that was not historically recorded before.Conclusions/significanceOur results indicate that when humans create novel habitats and ecological heterogeneities, they can provide evolutionary opportunities for rapid adaptive niche shifts associated with lineage divergence, whose consequences upon malaria transmission might be significant.

The poor management of the urban environment in sub-Saharan Africa is affecting Anopheles gambiae susceptibility to insecticides. A study was undertaken to assess the influence of breeding sites physicochemical parameters on malaria vectors population tolerance to insecticides. A total of 18, 262 larvae collected from 104 breeding sites were exposed to diagnostic concentrations of permethrin and deltamethrin. Larvae originating from cultivated sites were more tolerant than larvae from polluted or nonpolluted sites. No significant difference was observed between polluted and nonpolluted sites. Field larvae were 142 to 325 times and 6.08 to 9.57 times more tolerant to deltamethrin and permethrin, respectively, than larvae of the A. gambiae Kisumu strain used as control. A low but significant correlation was detected between physicochemical parameters and larval insecticide tolerance. Cultivated sites were negatively and significantly correlated to larval tolerance to both deltamethrin (r=−0.421; P

It has long been the goal of vector biology to generate genetic knowledge that can be used to "manipulate" natural populations of vectors to eliminate or lessen disease burden. While long in coming, progress towards reaching this goal has been made. Aiming to increase our understanding regarding the interactions between Plasmodium and the Anopheles immune genes, we investigated the patterns of genetic diversity of four anti-Plasmodium genes in the Anopheles gambiae complex of species.Within a comparative phylogenetic and population genetics framework, the evolutionary history of four innate immunity genes within the An. gambiae complex (including the two most important human malaria vectors, An. gambiae and An. arabiensis) is reconstructed. The effect of natural selection in shaping the genes' diversity is examined. Introgression and retention of ancestral polymorphisms are relatively rare at all loci. Despite the potential confounding effects of these processes, we could identify sites that exhibited dN/dS ratios greater than 1.In two of the studied genes, CLIPB14 and FBN8, several sites indicated evolution under positive selection, with CLIPB14 exhibiting the most consistent evidence. Considering only the sites that were consistently identified by all methods, two sites in CLIPB14 are adaptively driven. However, the analysis inferring the lineage -specific evolution of each gene was not in favor of any of the Anopheles lineages evolving under the constraints imposed by positive selection. Nevertheless, the loci and the specific amino acids that were identified as evolving under strong evolutionary pressure merit further investigation for their involvement in the Anopheles defense against microbes in general.