Low frequency of knockdown resistance mutation (L1014F) and the efficacy of PBO synergist in multiple insecticide-resistant populations of Anopheles gambiae in Ikorodu, Lagos State, Nigeria.

Objective: We evaluated the susceptibility status of Anopheles gambiae in two communities of Ikorodu, Lagos, Nigeria to DDT, deltamethrin, lambda cyhalothrin and bendiocarb. Methods: Anopheles immature stages were collected from their habitats in the surveyed community and allowed to emerge before exposure adult females to discriminating doses of WHO insecticides including DDT, deltamethrin, lambda cyhalothrin, bendiocarb and malathion. PBO synergistic bioassay was conducted for insecticides where the mosquito samples showed resistance. PCR assay was used for the detection of kdr mutation in the mosquitoes. Results: Resistance to DDT (40% and 86%) and lambda cyhalothrin (75% and 84%) in Oke-Ota and Majidun respectively. Suspected resistance to deltamethrin (94.9%) and bendiocarb (93.5%) was recorded in Oke-Ota community and the mosquitoes were susceptible to malathion in both communities. KDR mutation (L1014F) from resistance samples from both locations though with a low frequency that significantly departs from Hardy-Weinberg's probability (P> 0.01). PBO synergized bioassay was able to increase knockdown, percentage mortality and restore full susceptibility to deltamethrin and bendiocarb. Conclusion: Results from this study indicates that the metabolic resistance mechanism is highly implicated in the resistance to different classes of insecticide in Ikorodu and this should be taken into consideration when implementing vector control activities in this area.

Spatial distribution and ecological niche modeling of geographical spread of Anopheles gambiae complex in Nigeria using real time data.

The need for evidence-based data, to inform policy decisions on malaria vector control interventions in Nigeria, necessitated the establishment of mosquito surveillance sites in a few States in Nigeria. In order to make evidence-based-decisions, predictive studies using available data becomes imperative. We therefore predict the distribution of the major members of the Anopheles gambiae s.l. in Nigeria. Immature stages of Anopheles were collected from 72 study locations which span throughout the year 2020 resulted in the identification of over 60,000 Anopheline mosquitoes. Of these, 716 breeding sites were identified with the presence of one or more vector species from the An. gambiae complex and were subsequently used for modelling the potential geographical distribution of these important malaria vectors. Maximum Entropy (MaxEnt) distribution modeling was used to predict their potentially suitable vector habitats across Nigeria. A total of 23 environmental variables (19 bioclimatic and four topographic) were used in the model resulting in maps of the potential geographical distribution of three dominant vector species under current climatic conditions. Members of the An. gambiae complex dominated the collections (98%) with Anopheles stephensi, Anopheles coustani, Anopheles funestus, Anopheles moucheti, Anopheles nilli also present. An almost equal distribution of the two efficient vectors of malaria, An. gambiae and Anopheles coluzzii, were observed across the 12 states included in the survey. Anopheles gambiae and Anopheles coluzzii had almost equal, well distributed habitat suitability patterns with the latter having a slight range expansion. However, the central part of Nigeria (Abuja) and some highly elevated areas (Jos) in the savannah appear not suitable for the proliferation of these species. The most suitable habitat for Anopheles arabiensis was mainly in the South-west and North-east. The results of this study provide a baseline allowing decision makers to monitor the distribution of these species and establish a management plan for future national mosquito surveillance and control programs in Nigeria.

Genetic identity, human blood indices, and sporozoite rates of malaria vectors in Gaa-Bolorunduro, Kwara State, Nigeria.

INTRODUCTION: To identify the specific Anopheles mosquito sibling species responsible for malaria transmission, determine their vectorial potential, and predict suitable control measures, this study investigated genetic identities, human blood feeding, and sporozoite infection rates of endophilic Anopheles mosquitoes in Gaa-Bolorunduro, a cattle rearing community in Kwara State, Nigeria. METHODOLOGY: Monthly pyrethrum spray collections of Anopheles mosquitoes were conducted for one year in addition to PCR characterization of sibling species and ELISA probing of human blood meal and sporozoite infections. Mean numbers and human blood indices (HBI) of the different Anopheles sibling species identified were compared. RESULTS: The total of 668 PCR-identified mosquitoes comprised 50.8% An. arabiensis, 46.7% An. gambiae, and 2.5% An. coluzzii. Annual mean numbers of An. arabiensis was significantly higher (p = 0.001) than An. coluzzii but not An. gambiae (p = 0.602). Proportions of An. arabiensis found with human blood (0.29) were lower compared to An. gambiae (0.72) and An. coluzzii (0.75). However, the annual mean HBI of An. arabiensis was not significantly higher than An. gambiae (p = 0.195) and An. coluzzii (p = 0.249). Plasmodium falciparum sporozoite infection rate was 1.6% in An. gambiae, 0.9% in An. arabiensis and 0% in An. coluzzii. CONCLUSIONS: The prevalent An. arabiensis and An. gambiae mosquitoes found indoors, despite the outdoor cattle population barrier, could be targeted by community-scale utilization of long-lasting insecticide-treated bed nets. Further studies on outdoor mosquito surveillance and bovine blood meal identification are required for the recommendation of suitable complementary vector control measures for the community.

Pyrethroid Susceptibility in Culex quinquefasciatus Say. (Diptera: Culicidae) Populations from Delta State, Niger-Delta Region, Nigeria.

The development of insecticide resistance in different species of mosquitoes to Pyrethroids is a major challenge for vector-borne diseases transmitted by mosquitoes. Failure of Pyrethroids in control of mosquitoes would impact negatively on the gains recorded in control of mosquito-borne diseases in previous years. In anticipation of a country-wide deployment of Pyrethroid-treated nets for control of mosquito-borne diseases in Nigeria, this study assessed susceptibility of Culex quinquefasciatus Say. (Diptera: Culicidae) to Pyrethroids in Owhelogbo, Ejeme and Oria-Abraka communities in Delta State, Niger-Delta, Nigeria. Three to five day old Cx. quinquefasciatus were exposed to Deltamethrin (0.05%), Permethrin (0.75%), and Alphacypermethrin (0.05%) using World Health Organization bioassay method. Polymerase chain reaction (PCR) was employed in characterization of species and knockdown mutation. Results revealed that Cx. quinquefasciatus were generally susceptible (98-100%) to Deltamethrin, Permethrin, and Alphacypermethrin in the three communities with the exception of Owhelogbo where resistance to Deltamethrin (97%) was suspected. Knockdown time to Deltamethrin (11.51, 11.23, and 12.68 min), Permethrin (28.75, 13.26, and 14.49 min), and Alphacypermethrin (15.07, 12.50, and 13.03 min) were considerably low for Owhelogbo, Ejeme, and Oria-Abraka Cx. quinquefasciatus populations, respectively. Species identification result showed that all amplified samples were Cx. quinquefasciatus; however, no kdr allele was found in the three populations. Deployment of pyrethroid-treated nets for control of mosquito-borne diseases in Niger-Delta region of Nigeria is capable of reducing burden of diseases transmitted by Cx. quinquefasciatus as well as addressing nuisance value of the vector; however, caution must be entertained so as not to increase selection pressure thereby aiding resistance development.

Detection and Co-occurrence of kdr (F1534C and S989P) Mutations in Multiple Insecticides Resistant Aedes aegypti (Diptera: Culicidae) in Nigeria.

The outbreak of yellow fever virus transmitted by Aedes aegypti has been of major concern in Nigeria, this mosquito also transmits several other arboviruses globally. The control of many vectors of mosquito-borne diseases relies heavily on the use of insecticides. Therefore, constant monitoring of insecticide resistance status and associated mechanisms is crucial within the local mosquito population. Here, we determined the resistance profile of adult Ae. aegypti from Majidun and Oke Ota communities, Ikorodu Local Government Area of Lagos State, Nigeria to different classes of insecticides using WHO procedures. The resistant phenotypes of Ae. aegypti were screened for the presence of kdr mutations F1534C, S989P, and V1016G, which have been implicated in insecticide resistance in yellow fever vectors. A high level of resistance to DDT and pyrethroids was recorded in Ae. aegypti in this study, although possible resistance to deltamethrin, one of the pyrethroids was reported in one of the locations. Resistance to bendiocarb was recorded in the Majidun community while Ae. aegypti in both locations were susceptible to malathion. The presence of F1534C mutation associated with DDT and deltamethrin resistance in Ae. aegypti population, and the presence of S989P mutation were detected singly and in co-occurrence with F1534C for the first time in Africa, while V1016G mutation was not detected in this study. The role of these mutations in resistance phenotype expressed in Ae. aegypti in larger populations needs to be established.

Cytochrome P450 Mono-Oxygenase and Resistance Phenotype in DDT and Deltamethrin-Resistant Anopheles gambiae (Diptera: Culicidae) and Culex quinquefasciatus in Kosofe, Lagos, Nigeria.

Pyrethroids and DDT are key insecticides in the control of malaria, yellow fever, and lymphatic filariasis vectors. Knockdown and metabolic resistance mechanisms have been proven to be important in determining the efficacy of insecticides. Here we investigated cytochrome P450 as a resistance mechanism in Anopheles gambiae Giles and Culex quinquefasciatus Say exposed to deltamethrin and DDT. Two- to three-days-old adult female mosquitoes were used for insecticide exposures and PBO synergistic assays using WHO standard guidelines, kits and test papers (DDT 4%, deltamethrin 0.05%, and PBO 4%). Polymerase chain reaction (PCR) assays were used for the identification of the species and for characterization of the kdr allele. Mortality at 24 h post-exposure was 18 and 17% in An. gambiae s.s. exposed to DDT and deltamethrin, respectively; 1 and 5% in Cx. quinquefasciatus exposed to DDT and deltamethrin respectively. Significant (P < 0.01) levels of susceptibility was recorded in mosquitoes pre-exposed to PBO, as KDT50 and 24 h of exposure ranged from 37.6 min to 663.4 min and 27 to 80%, respectively. Presence of a knockdown resistance allele was recorded in An. gambiae s.s., 22.5% for homozygote resistance and 7.5% for heterozygotes, while Cx. quinquefasciatus populations showed no kdr allele despite the high level of resistance to DDT and deltamethrin. Findings from this study indicated that cytochrome P450 mono-oxygenase expression is highly implicated in the resistance phenotype to DDT and pyrethroids in An. gambiae and Cx. quinquefasciatus in the study area.

Pyrethroids resistance intensity and resistance mechanisms in Anopheles gambiae from malaria vector surveillance sites in Nigeria.

Anopheles gambiae, An. coluzzii and An. arabiensis are the three major vectors of malaria in Nigeria. These mosquitoes have developed resistance to different insecticides. Insecticides resistance intensity assay was recently introduced to provide insight into the potential operational significance of insecticide resistance. Here, we present data on pyrethroids resistance intensity and resistance mechanisms from six vector surveillance sites (Lagos, Ogun, Edo, Anambra, Kwara and Niger) in Nigeria. Adult Anopheles reared from larval collections were tested using WHO insecticides susceptibility protocol with 1x concentration of permethrin and deltamethrin followed with intensity assays with 5x and 10x concentrations of both insecticides. Synergistic and biochemical assays were carried out and underlying resistance mechanisms determined following standard protocols. Anopheles gambiae constituted >50% samples tested in five sites. Permethrin and deltamethrin resistance was observed at all the sites. The Kdt50 varied from 15 minutes (CI = 13.6-17.2) in deltamethrin to 42.1 minutes (CI = 39.4-44.1) in permethrin. For both insecticides, Kdt95 was >30 minutes with 25% to 87% post exposure mortality at the different sites. The West Africa knock down resistance (kdr-w) mechanism was found at each site. Resistant An. gambiae from Lagos, Ogun and Niger synergized prior to permethrin or deltamethrin exposure showed significant mortality (89-100%) compared to unsynergized mosquitoes (Lagos, p = 0.031; Ogun, p = 0.025; Niger, p = 0.018). Biochemical analyses revealed significant increased levels of P450 enzymes in resistant Anopheles gambiae from Lagos (p = 0.038); Ogun (p = 0.042) and Niger (p = 0.028) in addition to GST in Lagos (p = 0.028) and Ogun (p = 0.033). Overall, the results revealed high pyrethroid resistance associated with increased activities of metabolic enzymes (P450 + GST) in An. gambiae and An. coluzzii from Lagos and Ogun. The presence of kdr + P450 conferred moderate resistance whereas low resistance was the case where kdr was the sole resistance mechanism. Findings thus suggests that elevated levels of cytochrome P450 enzymes together with GST were responsible for high or severe pyrethroid resistance.

Neurogenomics in Africa: Perspectives, progress, possibilities and priorities.

The understanding of the genetic basis of neurological disorders has grown rapidly in the last two decades. Despite the genomic heterogeneity within African populations, large-scale candidate gene or linkage and exome studies are lacking. However, current knowledge on neurogenetics in African populations is limited and geographically very uneven. Isolated reports indicate the existence of autosomal dominant or recessive conditions incorporating cerebrovascular, movement, neuromuscular, seizure and motor neuron disorders in Africans. In addition, few African families with neurodegenerative disorders associated with dementia have been characterized in North, West and South Africa. The current insurgency in genomic research triggered by among others the Human Health and Heredity (H3) Africa Initiative indicates that there are unique opportunities to advance our knowledge and understanding of the influence of genomic variation on the pattern, presentations and prognosis of neurological disorders in Africa. These have enormous potential to unmask novel genes and molecular pathways germane to the neurobiology of brain disorders. It would facilitate the development of novel diagnostics, preventative and targeted treatments in the new paradigm of precision medicine. Nevertheless, it is crucial to strike a balance between effective traditional public health strategies and personalized genome based care. The translational barriers can be overcome through robust stakeholder engagement and sustainable multilevel, multigenerational and multidisciplinary capacity building and infrastructural development for genomic medicine in Africa.