High cases of submicroscopic Plasmodium falciparum infections in a suburban population of Lagos, Nigeria.

BACKGROUND: Asymptomatic malaria parasites are significant sources of infections for onward malaria transmission. Conventional tools for malaria diagnosis such as microscopy and rapid diagnostic test kits (RDT) have relatively low sensitivity, hence the need for alternative tools for active screening of such low-density infections. METHODS: This study tested var acidic terminal sequence-based (varATS) quantitative polymerase chain reaction (qPCR) for screening asymptomatic Plasmodium falciparum infections among dwellers of a sub-urban community in Lagos, Nigeria. Clinically healthy participants were screened for malaria using microscopy, RDT and varATS qPCR techniques. Participants were stratified into three age groups: 1-5, 6-14 and > 14 years old. RESULTS: Of the 316 participants screened for asymptomatic malaria infection, 78 (24.68%) were positive by microscopy, 99 (31.33%) were positive by RDT and 112 (35.44%) by varATS qPCR. Participants aged 6-14 years had the highest prevalence of asymptomatic malaria, with geometric means of ~ 116 parasites/µL and ~ 6689 parasites/µL as detected by microscopy and varATS, respectively. CONCLUSION: This study has revealed high prevalence of asymptomatic malaria in the study population, with varATS detecting additional sub-microscopic infections. The highest concentration of asymptomatic malaria was observed among school-age children between 6 and 14 years old. A large-scale screening to identify other potential hotspots of asymptomatic parasites in the country is recommended.

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.

Urinary schistosomiasis in school aged children of two rural endemic communities in Edo State, Nigeria.

BACKGROUND: Urinary schistosomiasis is endemic in many rural communities of Nigeria and school aged children are mostly affected. A cross-sectional study was carried out to assess the prevalence and intensity of urinary schistosomiasis infection among 251 school aged children in two communities of Ovia South West LGA of Edo State, Nigeria, as well as their knowledge on the control/elimination measures. METHODS: Urine samples were collected and examined by microscopy using filtration technique. In addition, a questionnaire survey was conducted among school-aged children and health care providers, probing their knowledge, attitude and practices on on-going control activities. RESULTS: The prevalence of urinary schistosomiasis among the school-aged children was 65.3%. The prevalence was generally higher among females (68.8%) and children in the age groups 10-14 (69.9%). The intensity of infection ranged from 1 to 5044 (mean=449.8) eggs/10ml of urine with a higher proportion having heavy infections (76.8%, P<0.05). Water contact was attested by 123 (49.0%) of the children; of these 123, 74 (60.1%) were infected. The children's knowledge on urinary schistosomiasis was deficient. CONCLUSION: The high prevalences reported in these communities require integrated approach to control which essentially should incorporate the provision of safe water supply and sanitary facilities, and health education in addition to the annual mass praziquantel distribution, to reduce transmission.

In vivo anti-plasmodial activities and toxic impacts of lime extract of a combination of Picralima nitida, Alstonia boonei and Gongronema latifolium in mice infected with Chloroquine-sensitive Plasmodium berghei.

BACKGROUND: Lime extracts of powdered combination of seeds of Picralima nitida, stem bark of Alstonia boonei and leaves of Gongronema latifolium is a common remedy used in the treatment of malaria in South Western Nigeria. OBJECTIVE: To determine the antiplasmodial activities of the combined herbal extracts and its impact on the haematological, hepatological and renological parameters in mice. METHODS: The 4-day suppressive and curative tests were used to assess the antiplasmodial activities of the extract in mice infected with chloroquine-sensitive Plasmodium berghei at concentration of 200mg/kg, 400mg/kg and 800mg/kg body weight. The haematological parameters including red blood cells, white blood cells, packed cell volume and haemoglobin count were analysed with an auto analyser. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were determined, while urea, protein and creatinine were analysed by standard procedural methods. RESULTS: The 4-day suppressive test revealed that the test extract achieved percentage suppression of 39.0%, 41.6% and 54.68% for the 200mg/kg, 400mg/kg and 800mg/kg concentration respectively. Additionally, the curative test achieved a high percentage suppression of 80.97%, 83.84% and 86.16% at the 200mg/kg, 400mg/kg and 800mg/kg concentration respectively. The extracts did not induce significant change on haematological parameters (P>0.05), while significant elevation in the values of the ALT and AST (P<0.05) was observed and elevation of creatinine (P<0.05) at 800mg/kg. CONCLUSIONS: The results support the traditional use of the herbal combination in the treatment of malaria, however the liver cells were impacted by the extracts in bioassay conducted with mice.

Microsatellite markers reveal low levels of population sub-structuring of Plasmodium falciparum in southwestern Nigeria.

BACKGROUND: Genetic diversity studies provide evidence of Plasmodium falciparum differentiation that could affect fitness and adaptation to drugs and target antigens for vaccine development. This study describes the genetic structure of P. falciparum populations in urban and rural sites from southwestern Nigeria. METHODOLOGY: Ten neutral microsatellite loci were genotyped in 196 P. falciparum infections from three localities: Aramoko-Ekiti, a rural community; Lekki, an urban location and Badagry, a peri-urban border settlement. Analysis was performed on the genetic diversity, linkage disequilibrium, population structure and inter-population differentiation. RESULTS: Allelic diversity values were similar across all populations, with mean expected heterozygosity (HE) values between 0.65 and 0.79. No matching multilocus haplotypes were found and analysis of multilocus LD showed no significant index of association. Genetic differentiation between populations was low (ΦPT = 0.017). CONCLUSION: The absence of detectable population structure of P. falciparum in southwestern Nigeria is evident in the lack of significant differentiation between populations separated by about 200 km. This implies that a fairly uniform malaria control strategy may be effective over a wide geographic range in this highly endemic region. However, more wide-scale survey across the country will be required to inform malaria control in this large and densely populated endemic region.

Abundance and diversity of Anopheles species (Diptera: Culicidae) associated with malaria transmission in human dwellings in rural and urban communities in Oyo State, Southwestern Nigeria.

Mosquito samples were collected from rural and urban communities in three selected major towns in Southwestern Nigeria to determine the impact of urbanization on the diversity and abundance of Anopheles species associated with malaria transmission in human habitations. A total of ten Anopheles species were identified in the rural communities, while eight Anopheles species were identified in the urban communities. Out of the ten Anopheles species identified, only four species, Anopheles gambiae (Giles), Anopheles funestus (Giles), Anopheles moucheti (Evans), and Anopheles nili (Theobald), were established to be vectors of malaria occurring in greater than 50% of the rural communities. Only A. gambiae occurred in all the urban communities, while the other three major vectors occurred in not more than 20% of the urban communities. Margalef's and Shannon-Wiener indices showed that diversity and species richness were higher in the rural compared to the urban. Comprehensive information on malaria vector abundance and diversity in rapidly changing communities is an important tool in planning and implementing successful vector control programs.

Evidence of carbamate resistance in urban populations of Anopheles gambiae s.s. mosquitoes resistant to DDT and deltamethrin insecticides in Lagos, South-Western Nigeria.

BACKGROUND: Resistance monitoring is essential in ensuring the success of insecticide based vector control programmes. This study was carried out to assess the susceptibility status of urban populations of Anopheles gambiae to carbamate insecticide being considered for vector control in mosquito populations previously reported to be resistant to DDT and permethrin. METHODS: Two - three day old adult female Anopheles mosquitoes reared from larval collections in 11 study sites from Local Government Areas of Lagos were exposed to test papers impregnated with DDT 4%, deltamethrin 0.05% and propoxur 0.1% insecticides. Additional tests were carried out to determine the susceptibility status of the Anopheles gambiae population to bendiocarb insecticide. Members of the A. gambiae complex, the molecular forms, were identified by PCR assays. The involvement of metabolic enzymes in carbamate resistance was assessed using Piperonyl butoxide (PBO) synergist assays. The presence of kdr-w/e and ace-1R point mutations responsible for DDT-pyrethroid and carbamate resistance mechanisms was also investigated by PCR. RESULTS: Propoxur resistance was found in 10 out of the 11 study sites. Resistance to three classes of insecticides was observed in five urban localities. Mortality rates in mosquitoes exposed to deltamethrin and propoxur did not show any significant difference (P > 0.05) but was significantly higher (P < 0.05) in populations exposed to DDT. All mosquitoes tested were identified as A. gambiae s.s (M form). The kdr -w point mutation at allelic frequencies between 45%-77% was identified as one of the resistant mechanisms responsible for DDT and pyrethroid resistance. Ace-1R point mutation was absent in the carbamate resistant population. However, the possible involvement of metabolic resistance was confirmed by synergistic assays conducted. CONCLUSION: Evidence of carbamate resistance in A. gambiae populations already harbouring resistance to DDT and permethrin is a clear indication that calls for the implementation of insecticide resistance management strategies to combat the multiple resistance identified.