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.

In vivo antiplasmodial activities and acute toxicity assessment of two plant cocktail extracts commonly used among Southwestern Nigerians.

Discovering and developing the desired antimalarials continue to be a necessity especially due to treatment failures, drug resistance, limited availability and affordability of antimalarial drugs and costs especially in poor malarial endemic countries. This study investigated the efficacies of two plant cocktails; CtA and CtB, selected based on their traditional usage. Efficacies of the cocktail extracts, chloroquine and pyrimethamine against Plasmodium berghei berghei were evaluated in mice using the suppressive, curative and prophylactic test models, after oral and intraperitoneal acute toxicity determination of the plant cocktails in accordance with Lorke's method. Data was analyzed using SPSS software version 23.0 with level of significance set at P < 0.05. The median lethal dose was determined to be higher than 5000 mg/kg body weight orally for both CtA and CtB; and 316.23 mg/kg body weight intraperitoneally for CtA. Each cocktail exhibited high dose dependent Plasmodium berghei berghei inhibition which was 96.95% and 99.13% in the CtA800 mg/kg and CtB800 mg/kg doses in the curative groups respectively, 96.46% and 78.62% for CtA800mg/kg and CtB800mg/kg doses in the suppressive groups respectively, as well as 65.05% and 88.80% for CtA800mg/kg and CtB800mg/kg doses in the prophylactic groups respectively. Throughout the observation periods, the standard drugs, chloroquine phosphate and pyrimethamine maintained higher inhibitions up to 100%. These findings demonstrate that CtA and CtB possess good antimalarial abilities and calls for their development and standardization as effective and readily available antimalarial options. The acute toxicity results obtained underscore the importance of obtaining information on toxicities of medicinal plant remedies before their administration in both humans and animals.

Knowledge, attitudes and perception of communities on mosquitoes and its control practices in Lagos State, Nigeria.

INTRODUCTION: evidence-based mosquito control strategy is important for efficient and effective delivery of mosquito control interventions. This is hinged on effective community participation and thorough understanding of the Knowledge Attitude and Practices (KAPs) to achieve desired result. Such community dynamics are often understudied. We designed this study to assess the perception of four local communities on aspects of mosquito behavior, prevention and control in Lagos State, Nigeria. METHODS: a cross-sectional survey was carried out using pretested semi-structured questionnaires to assess socio-demographic factors and KAPs in Kosofe, Alimosho, Ibeju-Lekki and Badagry Local Government Areas of Lagos State, Nigeria. Data analysis was carried out using IBM SPSS version 23. RESULTS: a total of 746 questionnaires were analyzed. Socio-demographic profile of the sampled population reveals that majority of the study population (73.1%) was between 18 and 40 years which constitute 49% males and 51% females. The knowledge of mosquito as a disease vector was high among the respondents which correlates with their level of education (P<0.05). The use of insecticide aerosols and Insecticides Treated Nets (ITNs) are the main control measures employed for mosquito control by respondents. Cost, convenience of usage and awareness majorly influenced the type of control measures that respondents adopt. Reasons such as not being easy to setup, skin irritation and the filling of being caged are reasons why some individuals do not use ITNs. Indoors, 32.4% of the respondents indicate the use of dichlorvos (DDVP) for household control of mosquitoes. CONCLUSION: the knowledge of mosquito control is high among middle aged individuals in Lagos State. Insecticide aerosols and ITNs are two major mosquito control methods used with DDVP insecticides frequently used indoors. This can inform the design of appropriate control methods in Lagos State.

Molecular detection of drug resistant polymorphisms in Plasmodium falciparum isolates from Southwest, Nigeria.

OBJECTIVE: Nigeria bears 25% of global malaria burden despite concerted efforts towards its control and elimination. The emergence of drug resistance to first line drugs, artemisinin combination therapies (ACTs), indicates an urgent need for continuous molecular surveillance of drug resistance especially in high burden countries where drug interventions are heavily relied on. This study describes mutations in Plasmodium falciparum genes associated with drug resistance in malaria; Pfk13, Pfmdr1, PfATPase6 and Pfcrt in isolates obtained from 83 symptomatic malaria patients collected in August 2014, aged 1-61 years old from South-west Nigeria. RESULTS: Two Pfmdr1, N86 and Y184 variants were present at a prevalence of 56% and 13.25% of isolates respectively. There was one synonymous (S679S) and two non-synonymous (M699V, S769M) mutations in the PATPase6 gene, while Pfcrt genotype (CVIET), had a prevalence of 45%. The Pfk13 C580Y mutant allele was suspected by allelic discrimination in two samples with mixed genotypes although this could not be validated with independent isolation or additional methods. Our findings call for robust molecular surveillance of antimalarial drug resistance markers in west Africa especially with increased use of antimalarial drugs as prophylaxis for Covid-19.

First report of AChE1 (G119S) mutation and multiple resistance mechanisms in Anopheles gambiae s.s. in Nigeria.

Susceptibility and PBO synergist bioassays were done using 3-5 days old female Anopheles mosquito collected from Lagos State, Nigeria with WHO test papers DDT (4%), permethrin (0.75%), Bendiocarb (1%) and PBO (4%) according to standard procedures. The activities of cytochrome P450s, glutathione S-transferase and carboxylesterases were determined using biochemical assays. The presence of kdr-w, kdr-e and Ace-1R mutations were examined using molecular assays. Resistance to DDT and permethrin in An gambiae s.s from the four Local Government Areas (LGAs) was recorded while suspected resistance to bendiocarb was recorded in mosquitoes from Alimosho and Kosofe LGAs. PBO synergist reduced the knockdown time and also recorded significantly (P < 0.05) higher 24 hrs percentage mortality compared to non-synergized bioassays. Increased activities of detoxifying enzymes was recorded in wild mosquito compared to the insecticides susceptible laboratory strain and this was significant (P < 0.05) in P450s, esterase α and ß. Kdr-w was detected in An. gambiae s.s from all the LGAs, kdr-e (L1014S) was detected in Alimosho, Kosofe and Ibeju-Lekki, while the Ace-1R gene was detected in Alimosho and Kosofe. Results from this study provide evidence for resistance of An. gambiae from Lagos State to multiple classes of neurotoxic insecticides with multiple resistance mechanisms to these insecticides.

A barcode of multilocus nuclear DNA identifies genetic relatedness in pre- and post-Artemether/Lumefantrine treated Plasmodium falciparum in Nigeria.

BACKGROUND: The decline in the efficacy of artemisinin-based combination treatment (ACT) in some endemic regions threatens the progress towards global elimination of malaria. Molecular surveillance of drug resistance in malaria-endemic regions is vital to detect the emergence and spread of mutant strains. METHODS: We observed 89 malaria patients for the efficacy of artemether-lumefantrine for the treatment of uncomplicated Plasmodium falciparum infections in Lagos, Nigeria and determined the prevalence of drug resistant strains in the population. Parasite clearance rates were determined by microscopy and the highly sensitive var gene acidic terminal sequence (varATS) polymerase chain reaction for 65 patients with samples on days 0, 1, 3, 7, 14, 21 and 28 after commencement of treatment. The genomic finger print of parasite DNA from pre- and post-treatment samples were determined using 24 nuclear single nucleotide polymorphisms (SNP) barcode for P. falciparum. Drug resistance associated alleles in chloroquine resistance transporter gene (crt-76), multidrug resistance genes (mdr1-86 and mdr1-184), dihydropteroate synthase (dhps-540), dihydrofolate reductase (dhfr-108) and kelch domain (K-13580) were genotyped by high resolution melt analysis of polymerase chain reaction (PCR) fragments. RESULTS: By varATS qPCR, 12 (18.5%) of the participants had detectable parasite DNA in their blood three days after treatment, while eight (12.3%) individuals presented with genotypable day 28 parasitaemia. Complexity of infection (CoI) was 1.30 on day 0 and 1.34 on day 28, the mean expected heterozygosity (HE) values across all barcodes were 0.50 ± 0.05 and 0.56 ± 0.05 on days 0 and 28 respectively. Barcode (π) pairwise comparisons showed high genetic relatedness of day 0 and day 28 parasite isolates in three (37.5%) of the eight individuals who presented with re-appearing infections. Crt-76 mutant allele was present in 38 (58.5%) isolates. The mdr1-86 mutant allele was found in 56 (86.2%) isolates. No mutation in the K-13580 was observed. CONCLUSIONS: Persistence of DNA-detectable parasitaemia in more than 18% of cases after treatment and indications of genetic relatedness between pre- and post-treatment infections warrants further investigation of a larger population for signs of reduced ACT efficacy in Nigeria.

Toxicological effects of prolonged and intense use of mosquito coil emission in rats and its implications on malaria control.

Mosquito coil is a vector control option used to prevent malaria in low income counties, while some studies have addressed this issue, additional reseach is required to increase knowledge on the adverse health effects caused by the prolonged use of coils. In this study we investigated the toxicological effects of fumes from two locally manufactured mosquito coil insecticides (with pyrethroids: transfluthrin and d-allethrin as active ingredients) on male albino rats. For this, we recorded the haematological and biochemical indices, and made histopathology and mutagenicity evaluations in rats exposed to mosquito fumes during 2, 4, 8, 12 and 16 week periods. Haematological determination was performed using automated hematology analyzer to determine White Blood Cell (WBC), Packed Cell Volume (PCV), Red Blood Cell (RBC) and Platelet (PLT) counts, while biochemical evaluations were determined using available commercial kits. Gross histopathological changes were studied for the kidney, liver and lungs in sacrificed rats. The rat sperm head abnormalities assessment was used to evaluate mutagenicity. Mosquito coil fumes produced significant increase (P < 0.05) in the levels of total protein, total albumin and bilirubin, when animals were exposed from two weeks to 16 weeks with transfluthrin. Similarly, elevation in the activities of aspartate amino transferase, alanine amino transferase and alanine phosphatase, increased significantly in both insecticides. Increase in WBC, RBC and PCV were recorded for all the exposure periods, however PLT count showed no significant increase (P > 0.05). Mutagenicity assessment revealed sperm abnormality was statistically significant (P < 0.05) compared with the control at 8, 12 and 16 weeks post exposure to transfluthrin. Histological studies revealed severe lung damage evidenced by interstitial accumulations, pulmonary oedema and emphysema in exposed rats. Intracellular accumulations and severe sinusoidal congestion of liver cells were observed from 12 weeks exposure, indicating liver damage. Our studies indicate that mosquito coil fumes do initiate gradual damage to the host. These pathological effects must be taken into consideration by the malaria control program, particularly when regulating their long term and indoor usage.