Highlights on Anopheles nili and Anopheles moucheti, Malaria Vectors in Africa

In this chapter, we review knowledge gained so far on mosquitoes from An. moucheti and closely related species, as well as the An. nili complex. We highlight specific bionomical, ecological and genetic attributes that distinguish these species from the most well-known major African malaria vectors, providing opportunities for further research on neglected aspects of vector biology and control

Awareness, attitudes and prevention of malaria in the cities of Douala and Yaoundé (Cameroon)

<p>Abstract</p> <p>Background</p> <p>There is little information on the social perception of malaria and the use of prevention methods in Cameroon. This study was designed to assess knowledge, attitude and management of malaria in households living in the cities of Douala and Yaoundé.</p> <p>Results</p> <p>Over 82% of people interviewed associated malaria transmission to mosquito bites. Methods used for malaria prevention were: environmental sanitation 1645 (76.1%), use of bed nets 1491 (69%), insecticide spray/coils 265 (12.3%) and netting of doors or windows 42 (1.9%). Bed net ownership was significantly high in Yaoundé (73.8%) (P < 0.0001), whereas the use of insecticide spray or coils was significantly important in Douala (16.3%) (P < 0.0001). Some of the problems experienced by families using ITN were the difficulty in finding chemicals for the retreatment of nets 702 (47%), insufficient financial means to buy new bed nets to replace old ones 366 (24.5%) or, to provide bed nets to everybody in the household 289 (19.4%) and the sensation of feeling excessive heat when sleeping under a bed net 74 (5%). The amount spent monthly by a household for vector control and malaria treatment was estimated at 2377 fcfa (3.6 euros) and 4562 fcfa (6.95 euros) respectively. These amounts were not significantly different between households of Douala and Yaoundé. Concerning management of malaria cases, 18.6% of people declare going to the hospital when suffering from malaria. The majority of people (81.4%) do self medication - they either buy drugs from the pharmacists, street sellers or they use plants to cure malaria.</p> <p>Conclusion</p> <p>The study revealed a high awareness of populations on malaria and ITNs. However some attitudes hindering the use of ITN or related to the management of clinical cases need further attention.</p

Chikungunya outbreak in a rural area of Western Cameroon in 2006: A retrospective serological and entomological survey

<p>Abstract</p> <p>Background</p> <p>Although arboviral infections including Chikungunya virus (CHIKV) are common in sub-Saharan Africa, data on their circulation and prevalence are poorly documented. In 2006, more than 400 cases of dengue-like fever were reported in Kumbo (Northwest Region of Cameroon). The aim of this study was to identify the aetiology of this fever and to define its extent in the area.</p> <p>Methods</p> <p>We conducted a cross-sectional seroprevalence survey one year after clinical investigations to define the extent of the infection. An entomological survey consisted of the collection and identification of mosquito immature stages in water containers in or around human dwellings.</p> <p>Results</p> <p>A total of 105 sera were obtained from volunteers and tested for CHIKV, O'Nyong-nyong virus (ONNV) and Dengue virus (DENV) specific IgM and IgG antibodies by enzyme-linked immunosorbent assays (ELISA). CHIKV infection was defined as the presence of IgM antibodies to CHIKV. There was serological evidence for recent Chikungunya infection, as 54 subjects (51.4%) had detectable IgM anti-CHIKV in their sera. Amongst these, 52 showed both anti-CHIKV IgM and IgG, and 2 (1.9%) had IgM anti-CHIKV in the absence of IgG. Isolated anti-CHIKV IgG positives were detected in 41 (39%) cases. No anti-ONNV and anti-DENV IgM antibodies were found amongst the sample tested. Out of 305 larvae collected in the different breeding sites, 87 developed to the adult stage; 56 (64.4%) were <it>Aedes africanus </it>and the remaining <it>Culex </it>spp.</p> <p>Conclusions</p> <p>These findings suggest that the outbreak of febrile illness reported in three villages of Western Cameroon was due to CHIKV. The issue of a possible persistence of anti-CHIKV IgM antibodies is discussed. <it>Ae. africanus </it>which was found to be relatively abundant among the raffia palm bushes probably plays a role in the transmission of CHIKV along the chain of sylvatic/domestic mosquito species in this rural area. Particular attention should therefore be given to arbovirus infections in the Central African sub-region where these infections are becoming an emerging public health threat.</p

Improving the population genetics toolbox for the study of the African malaria vector Anopheles nili: microsatellite mapping to chromosomes

<p>Abstract</p> <p>Background</p> <p><it>Anopheles nili </it>is a major vector of malaria in the humid savannas and forested areas of sub-Saharan Africa. Understanding the population genetic structure and evolutionary dynamics of this species is important for the development of an adequate and targeted malaria control strategy in Africa. Chromosomal inversions and microsatellite markers are commonly used for studying the population structure of malaria mosquitoes. Physical mapping of these markers onto the chromosomes further improves the toolbox, and allows inference on the demographic and evolutionary history of the target species.</p> <p>Results</p> <p>Availability of polytene chromosomes allowed us to develop a map of microsatellite markers and to study polymorphism of chromosomal inversions. Nine microsatellite markers were mapped to unique locations on all five chromosomal arms of <it>An. nili </it>using fluorescent <it>in situ </it>hybridization (FISH). Probes were obtained from 300-483 bp-long inserts of plasmid clones and from 506-559 bp-long fragments amplified with primers designed using the <it>An. nili </it>genome assembly generated on an Illumina platform. Two additional loci were assigned to specific chromosome arms of <it>An. nili </it>based on <it>in silico </it>sequence similarity and chromosome synteny with <it>Anopheles gambiae</it>. Three microsatellites were mapped inside or in the vicinity of the polymorphic chromosomal inversions <it>2Rb </it>and <it>2Rc</it>. A statistically significant departure from Hardy-Weinberg equilibrium, due to a deficit in heterozygotes at the <it>2Rb </it>inversion, and highly significant linkage disequilibrium between the two inversions, were detected in natural <it>An. nili </it>populations collected from Burkina Faso.</p> <p>Conclusions</p> <p>Our study demonstrated that next-generation sequencing can be used to improve FISH for microsatellite mapping in species with no reference genome sequence. Physical mapping of microsatellite markers in <it>An. nili </it>showed that their cytological locations spanned the entire five-arm complement, allowing genome-wide inferences. The knowledge about polymorphic inversions and chromosomal locations of microsatellite markers has been useful for explaining differences in genetic variability across loci and significant differentiation observed among natural populations of <it>An. nili</it>.</p

Multiple Insecticide Resistance in the Malaria Vector Anopheles funestus from Northern Cameroon Is Mediated by Metabolic Resistance Alongside Potential Target Site Insensitivity Mutations

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

Investigation of mechanisms of bendiocarb resistance in Anopheles gambiae populations from the city of Yaoundé, Cameroon

BACKGROUND Resistance to the carbamate insecticide bendiocarb is emerging in Anopheles gambiae populations from the city of Yaoundé in Cameroon. However, the molecular basis of this resistance remains uncharacterized. The present study objective is to investigate mechanisms promoting resistance to bendiocarb in An. gambiae populations from Yaoundé. METHODS The level of susceptibility of An. gambiae s.l. to bendiocarb 0.1 % was assessed from 2010 to 2013 using bioassays. Mosquitoes resistant to bendiocarb, unexposed and susceptible mosquitoes were screened for the presence of the Ace-1(R) mutation using TaqMan assays. Microarray analyses were performed to assess the pattern of genes differentially expressed between resistant, unexposed and susceptible. RESULTS Bendiocarb resistance was more prevalent in mosquitoes originating from cultivated sites compared to those from polluted and unpolluted sites. Both An. gambiae and Anopheles coluzzii were found to display resistance to bendiocarb. No G119S mutation was detected suggesting that resistance was mainly metabolic. Microarray analysis revealed the over-expression of several cytochrome P450 s genes including cyp6z3, cyp6z1, cyp12f2, cyp6m3 and cyp6p4. Gene ontology (GO) enrichment analysis supported the detoxification role of cytochrome P450 s with several GO terms associated with P450 activity significantly enriched in resistant samples. Other detoxification genes included UDP-glucosyl transferases, glutathione-S transferases and ABC transporters. CONCLUSION The study highlights the probable implication of metabolic mechanisms in bendiocarb resistance in An. gambiae populations from Yaoundé and stresses the need for further studies leading to functional validation of detoxification genes involved in this resistance

Influence of the agrochemicals used for rice and vegetable cultivation on insecticide resistance in malaria vectors in southern Côte d’Ivoire

Background Vector control can contribute to the development of resistance to insecticides in malaria vectors. As the swamps and wetlands used for some agricultural activities constitute productive breeding sites for many mosquito species, agricultural pest control may increase the selection pressure for insecticide resistance in mosquitoes. Understanding the use of agrochemicals by farmers is important to plan and initiate effective integrated pest and vector management interventions. Methods A knowledge–attitude–practice study, using questionnaires, was undertaken with 102 rice farmers in Tiassalé and 106 vegetable farmers in Dabou (South Côte d’Ivoire) in order to generate information on pesticide usage. In addition, insecticide susceptibility bioassays were conducted using adult mosquitoes obtained from larvae collected within farms, and the persistence of agricultural pesticides in the farming environment, including sediment and mosquito breeding site water, was investigated by HPLC. Results Herbicides and insecticides appeared to be the most frequently used pesticides for both crops. Amino phosphonates (mostly glyphosate) represented the most used herbicides (45 % for rice up to 89 % for vegetables). Pyrethroids appeared to be the most used insecticides (accounting for 90 % of all the insecticide use reported). Approximately 75 % of respondents had not been to school and do not understand product labels. Only about 45 % of farmers respect the recommended pesticide dosage and about 10–15 % of pesticides used for rice and vegetable, respectively, are not recommended for these crops. As per WHO criteria, the mosquito local populations from the two localities were resistant to three of the four insecticides tested, as mortalities were less than 35 % for deltamethrin, DDT and bendiocarb. Higher susceptibility was observed for malathion, although the population was considered resistant in Dabou (80 % mortality) and susceptible in Tiassalé (98 % mortality). With the exception of glyphosate, residues from each of six chemicals tested for were detected in each of the sites visited in the two localities. Conclusion The study describes the use of insecticides and herbicides on crops and highlights the importance of considering agriculture practices when attempting to manage resistance in malaria vectors. Inter-sectoral collaboration between agriculture and public health is required to develop efficient integrated pest and vector management interventions

Habitat and Seasonality Affect Mosquito Community Composition in the West Region of Cameroon.

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

Status of Insecticide Resistance and Its Mechanisms in and Populations from Forest Settings in South Cameroon.

A key factor affecting malaria vector control efforts in Cameroon is the rapid expansion of insecticide resistance in s.l ) populations; however, mechanisms involved in insecticide resistance in forest mosquito populations are still not well documented yet. The present study was conducted to screen molecular mechanisms conferring insecticide resistance in s.l. populations from the South Cameroon forest region. WHO bioassays were conducted with F0 females aged three to four days from forest (Sangmelima, Nyabessan, and Mbandjock) and urban sites (Yaoundé (Bastos and Nkolondom)), against pyrethroids (permethrin 0.75% and deltamethrin 0.05%) and carbamates (bendiocarb 0.1%). Members of the s.l. species complex were identified using molecular diagnostic tools. TaqMan assays were used to screen for target site mutations. The expression profiles of eight genes implicated in insecticide resistance were assessed using RT-qPCR. Cuticle hydrocarbon lipids were measured to assess their potential implication in insecticide resistance. Both and were detected. was highly prevalent in Sangmelima, Nyabessan, Mbandjock, and Nkolondom. was the only species found in the Yaoundé city center (Bastos). Low mortality rate to both pyrethroids and bendiocarb was recorded in all sites. High frequency of L1014F allele (75.32-95.82%) and low frequencies of L1014S (1.71-23.05%) and N1575Y (5.28-12.87%) were recorded. The G119S mutation (14.22-35.5%) was detected for the first time in populations from Cameroon. This mutation was rather absent from populations. The detoxification genes , , , , as well as which catalyzes epicuticular hydrocarbon biosynthesis, were found to be overexpressed in at least one population. The total cuticular hydrocarvbon content, a proxy of cuticular resistance, did not show a pattern associated with pyrethroid resistance in these populations. The rapid emergence of multiple resistance mechanisms in s.l. population from the South Cameroon forest region is of big concern and could deeply affect the sustainability of insecticide-based interventions strategies in this region

Population structure of the malaria vector Anopheles moucheti in the equatorial forest region of Africa

<p>Abstract</p> <p>Background</p> <p><it>Anopheles moucheti </it>is a major malaria vector in forested areas of Africa. However, despite its important epidemiological role, it remains poorly known and insufficiently studied. Here, levels of genetic differentiation were estimated between different <it>A. moucheti </it>populations sampled throughout its distribution range in Central Africa.</p> <p>Methods</p> <p>Polymorphism at ten microsatellite markers was compared in mosquitoes sampled in Cameroon, the Democratic Republic of Congo and an island on Lake Victoria in Uganda. Microsatellite data were used to estimate genetic diversity within populations, their relative long-term effective population size, and the level of genetic differentiation between them.</p> <p>Results</p> <p>All specimens collected in Tsakalakuku (Democratic Republic of Congo) were identified as <it>A. m. bervoetsi </it>while other samples consisted of <it>A. m. moucheti</it>. Successful amplification was obtained at all microsatellite loci within all <it>A. m. moucheti </it>samples while only six loci amplified in <it>A. m. bervoetsi</it>. Allelic richness and heterozygosity were high for all populations except the island population of Uganda and <it>A. m. bervoetsi</it>. High levels of genetic differentiation were recorded between <it>A. m. bervoetsi </it>and each <it>A. m. moucheti </it>sample as well as between the island population of <it>A. m. moucheti </it>and mainland populations. Significant isolation by distance was evidenced between mainland populations.</p> <p>Conclusion</p> <p>High levels of genetic differentiation supports complete speciation of <it>A. m. bervoetsi </it>which should henceforth be recognized as a full species and named <it>A. bervoetsi</it>. Isolation by distance is the main force driving differentiation between mainland populations of <it>A. m. moucheti</it>. Genetically and geographically isolated populations exist on Lake Victoria islands, which might serve as relevant field sites for evaluation of innovative vector control strategies.</p