Molecular Identification and Characterization of Plasmodium ovale curtisi in Field Isolates from Symptomatic Children in North-Central Nigeria.

PURPOSE: Plasmodium ovale is not usually the focus of most malaria research or intervention programmes and has lately been termed the neglected human malaria parasites. The parasite exists as two genetically distinct sympatric species namely P. ovale curtisi and P. ovale wallikeri but information on the distribution of P. ovale sub-species is lacking in Nigeria. The objective of this study, therefore, was aimed at characterizing the P. ovale sub-species in isolates from symptomatic individuals in North-central Nigeria. METHODS: Parasites were identified by light microscopy of Giemsa stained thick and thin blood films. Molecular characterization and confirmation of P. ovale sub-species were done by species-specific nested PCR and sequencing of the small subunit ribosomal RNA (SSUrRNA) gene. RESULTS: A total of 412 children were enrolled into this study of which 88.6% (n = 365) were positive for Plasmodium species by nested PCR and P. falciparum was predominant. Of the 365 isolates, 4 (1.1%) had P. ovale infections and of these, 3 (0.8%) were mixed species infections of P. ovale with P. falciparum. DNA sequence analysis confirmed that all the four P. ovale parasites were P. ovale curtisi as their sequences were 99-100% identical to previously published P. ovale curtisi sequences in the GenBank and they cluster with the P. ovale curtisi sequences by phylogeny. CONCLUSION: Our findings demonstrate the occurrence of P. ovale curtisi in the study area. This has implications for public health and malaria elimination programmes, since they also serve as potential risk to travellers from malaria-free regions.

Genetic diversity and complexity of Plasmodium falciparum infections in the microenvironment among siblings of the same household in North-Central Nigeria.

BACKGROUND: Plasmodium falciparum parasites are known to exhibit extensive genetic diversity in areas of high transmission intensity and infected individuals in such communities often harbour several complex mixtures of parasite clones with different genetic characteristics. However, in the micro-environment, the extent of genetic diversity of P. falciparum parasites remain largely unknown. In this study therefore, the complexity of P. falciparum infections in households was investigated among symptomatic siblings, living under the same roof in north-central Nigeria. METHODS: Children were enrolled into the study if they were at least two from a household and presented with symptoms of uncomplicated malaria. Clinical malaria was confirmed by light microscopy of Giemsa-stained thick and thin blood films. Genomic DNA was isolated from blood spots on filter paper. Molecular characterization of P. falciparum isolates was done by allele-specific nested PCR of the highly polymorphic merozoite surface protein-2 (msp-2) gene. RESULTS: Ninety-three children from 43 households were enrolled into this study. A total of 26 different msp-2 alleles were identified from 215 fragments (range: 180-480 bp). Majority of the isolates [65.6% (n = 61)] were polyclonal infections consisting of 2-6 clones and were significantly more common with the FC27 allelic family (p = 0.036). The multiplicity of infection (MOI) per household ranged from 1.0 to 4.5 while the overall MOI in the study population was 2.31. The pattern of distribution of msp-2 allele types among the households fell into two categories: households where both msp-2 allele types (FC27 and 3D7) were present; households where only one msp-2 allele type (FC27 or 3D7) was present. Majority of the households [88.4% (n = 38)], had both msp-2 allele types but they were disproportionately distributed among the children while in a few households [11.6% (n = 5)], all the children were infected with only one type of msp-2 allele. CONCLUSION: These findings showed that P. falciparum isolates exhibit remarkable degree of genetic diversity in the micro-environment and are composed mainly of multiclonal infections, which is an indication of a high ongoing parasite transmission. This suggests that the micro-environment is an important area of focus for malaria control interventions and for evaluating intervention programmes.

Safety of Artemisinin-Based Combination Therapies in Nigeria: A Cohort Event Monitoring Study.

BACKGROUND: A pilot programme of Cohort Event Monitoring (CEM) was conducted across the six geopolitical zones of Nigeria on patients treated for uncomplicated malaria with artemisinin-based combination therapy (ACT). The emergence and spread of malaria parasites resistant to commonly available antimalarial drugs necessitated a shift in policy for malaria treatment by the Federal Government from the use of chloroquine and sulphadoxine-pyrimethamine (SP) as first-line treatments to ACTs. Initial reports following deployment of ACTs in clinical settings raised safety concerns regarding their use. Although artemisinin and its derivatives are generally thought to be safe, there are currently few or no data on their safety among populations in Nigeria. OBJECTIVES: The main objectives of the CEM programme were to proactively determine the adverse event (AE) profile of artesunate/amodiaquine (AA) and artemether/lumefantrine (AL) in real-life settings and to find out the factors predisposing to AEs. METHODS: The CEM study was observational, longitudinal, prospective, and inceptional. Patients were observed in real-life situations. It was conducted in six public health facilities in Nigeria on patients with a clinical diagnosis of uncomplicated malaria treated with ACTs. Patients were prescribed one of the ACTs on an alternate basis as they enrolled into the programme. Follow-up reviews were undertaken on days 3 and 7 following commencement of ACT treatment. At follow-up, patients were evaluated for any clinical event that they might have experienced following the use of the ACTs. We report the result of this initial pilot in which 3,010 patients treated for uncomplicated malaria with AA or AL were enrolled. RESULTS: The seven most common AEs seen were general body weakness 25.0/36.6% (AL/AA); dizziness 11.9/17.2% (AL/AA); vomiting 8.0/10.2% (AL/AA); abdominal pain 8.5/7.2% (AL/AA); insomnia 6.3/5.9% (AL/AA); body pains 3.4/5.2 (AL/AA) %; anorexia 8.5/4.6% (AL/AA). Most adverse events occurred from day 1 and peaked by day 2 and 3 of medication with the mean duration of events being 3 days. By the end of the follow-up visit on day 7, the AEs had resolved in the majority of patients. Adverse events were more common in the AA group than AL revealing a better safety profile for AL (p < 0.001). Both ACTs demonstrated good ability to resolve the clinical symptoms of uncomplicated malaria. CONCLUSION: In conclusion, this pilot CEM programme suggests that adverse events with ACTs were common. However, serious life-threatening events were not common. It appears that ACTs have a tolerable safety profile among Nigerians.