Mbanga Muleba

High burden of malaria following scale-up of control interventions in Nchelenge District, Luapula Province, Zambia

BackgroundMalaria control interventions have been scaled-up in Zambia in conjunction with a malaria surveillance system. Although substantial progress has been achieved in reducing morbidity and mortality, national and local information demonstrated marked heterogeneity in the impact of malaria control across the country. This study reports the high burden of malaria in Nchelenge District, Luapula Province, Zambia from 2006 to 2012 after seven years of control measures.MethodsYearly aggregated information on cases of malaria, malaria deaths, use of malaria diagnostics, and malaria control interventions from 2006 to 2012 were obtained from the Nchelenge District Health Office. Trends in the number of malaria cases, methods of diagnosis, malaria positivity rate among pregnant women, and intervention coverage were analysed using descriptive statistics.ResultsMalaria prevalence remained high, increasing from 38% in 2006 to 53% in 2012. Increasing numbers of cases of severe malaria were reported until 2010. Intense seasonal malaria transmission was observed with seasonal declines in the number of cases between April and August, although malaria transmission continued throughout the year. Clinical diagnosis without accompanying confirmation declined from 95% in 2006 to 35% in 2012. Intervention coverage with long-lasting insecticide-treated nets and indoor residual spraying increased from 2006 to 2012.ConclusionsDespite high coverage with vector control interventions, the burden of malaria in Nchelenge District, Zambia remained high. The high parasite prevalence could accurately reflect the true burden, perhaps in part as a consequence of population movement, or improved access to care and case reporting. Quality information at fine spatial scales will be critical for targeting effective interventions and measurement of progress.

An Operational Framework for Insecticide Resistance Management Planning

Country-wide planning and coordination can improve sustainability of vectorborne disease control.

Underestimation of foraging behaviour by standard field methods in malaria vector mosquitoes in southern Africa

BackgroundDefining the anopheline mosquito vectors and their foraging behaviour in malaria endemic areas is crucial for disease control and surveillance. The standard protocol for molecular identification of host blood meals in mosquitoes is to morphologically identify fed mosquitoes and then perform polymerase chain reaction (PCR), precipitin tests, or ELISA assays. The purpose of this study was to determine the extent to which the feeding rate and human blood indices (HBIs) of malaria vectors were underestimated when molecular confirmation by PCR was performed on both visually fed and unfed mosquitoes.MethodsIn association with the Southern Africa International Centers of Excellence in Malaria Research (ICEMR), mosquito collections were performed at three sites: Choma district in southern Zambia, Nchelenge district in northern Zambia, and Mutasa district in eastern Zimbabwe. All anophelines were classified visually as fed or unfed, and tested for blood meal species using PCR methods. The HBIs of visually fed mosquitoes were compared to the HBIs of overall PCR confirmed fed mosquitoes by Pearson’s Chi-Square Test of Independence.ResultsThe mosquito collections consisted of Anopheles arabiensis from Choma, Anopheles funestus s.s., Anopheles gambiae s.s. and Anopheles leesoni from Nchelenge, and An. funestus s.s. and An. leesoni from Mutasa. The malaria vectors at all three sites had large human blood indices (HBI) suggesting high anthropophily. When only visually fed mosquitoes tested by PCR for blood meal species were compared to testing those classified as both visually fed and unfed mosquitoes, it was found that the proportion blooded was underestimated by up to 18.7%. For most Anopheles species at each site, there was a statistically significant relationship (P < 0.05) between the HBIs of visually fed mosquitoes and that of the overall PCR confirmed fed mosquitoes.ConclusionThe impact on HBI of analysing both visually fed and unfed mosquitoes varied from site to site. This discrepancy may be due to partial blood feeding behaviour by mosquitoes, digestion of blood meals, sample condition, and/or expertise of entomology field staff. It is important to perform molecular testing on all mosquitoes to accurately characterize vector feeding behaviour and develop interventions in malaria endemic areas.

Spatio-temporal heterogeneity of malaria vectors in northern Zambia: implications for vector control

BackgroundDespite large reductions in malaria burden across Zambia, some regions continue to experience extremely high malaria transmission. In Nchelenge District, Luapula Province, northern Zambia, almost half the human population carries parasites. Intervention coverage has increased substantially over the past decade, but comprehensive district-wide entomological studies to guide delivery of vector control measures are lacking. This study describes the bionomics and spatio-temporal patterns of malaria vectors in Nchelenge over a two and a half year period, investigates what household factors are associated with high vector densities and determines why vector control may not have been effective in the past to better guide future control efforts.MethodsBetween April 2012 and September 2014, twenty-seven households from across Nchelenge District were randomly selected for monthly light trap collections of mosquitoes. Anopheline mosquitoes were identified morphologically and molecularly to species. Foraging rates were estimated and sporozoite rates were determined by circumsporozoite ELISAs to calculate annual entomological inoculation rates. Blood feeding rates and host preference were determined by PCR. Zero-inflated negative binomial models measured environmental and household factors associated with mosquito abundance at study households such as season, proximity to the lake, and use of vector control measures.ResultsThe dominant species in Nchelenge was An. funestus (s.s.) with An. gambiae (s.s.) as a secondary vector. Both vectors were found together in large numbers across the district and the combined EIRs of the two vectors exceeded 80 infectious bites per person per annum. An. funestus household densities increased in the dry season whilst An. gambiae surged during the rains. Presence of insecticide treated nets (ITNs) and closed eaves in the houses were found to be associated with fewer numbers of An. gambiae but not An. funestus. There was no association with indoor residual spraying (IRS).ConclusionIn Nchelenge, the co-existence of two highly anthropophagic vectors, present throughout the year, is likely to be driving the high malaria transmission evident in the district. The vectors here have been shown to be highly resistant to pyrethroids used for IRS during the study. Vector control interventions in this area would have to be multifaceted and district-wide for effective control of malaria.

Benchmarking insecticide resistance intensity bioassays for Anopheles malaria vector species against resistance phenotypes of known epidemiological significance

BackgroundInsecticide use via indoor residual spraying (IRS) or treated nets is the primary method for controlling malaria vector populations. The incidence of insecticide resistance in vector populations is burgeoning globally making resistance management key to the design of effective malaria control and elimination strategies. Vector populations can be assessed for insecticide resistance using a binary (susceptible or resistant) classification based on the use of the standard WHO insecticide susceptibility assay for adult anopheline mosquitoes. However, the recent scaling up of vector control activities has necessitated a revision of the WHO bioassay protocol to include the production of information that not only diagnoses resistance but also gives information on the intensity of expression of resistance phenotypes detected. This revised protocol is expected to inform on the range of resistance phenotypes in a target vector population using discriminating/diagnostic insecticide concentrations (DC) as well as their potential operational significance using 5× DC and 10× DC assays. The aim of this project was to use the revised protocol to assess the intensity of pyrethroid resistance in a range of insecticide resistant Anopheles strains with known resistance mechanisms and for which there is evidence of operational significance in the field setting from which these colonies were derived.MethodsDiagnostic concentration (DC) bioassays followed by 5× DC and 10× DC assays using the pyrethroid insecticides permethrin and deltamethrin were conducted according to the standard WHO bioassay method against pyrethroid resistant laboratory strains of Anopheles funestus, An. arabiensis and An. gambiae.ResultsLow to moderate resistance intensities were recorded for the An. arabiensis and An. gambiae strains while moderate to high intensities were recorded for the An. funestus strains.ConclusionsIt is evident that resistance intensity assays can add predictive value to the decision making process in vector control settings, although more so in an IRS setting and especially when bench-marked against resistance phenotypes of known operational significance.

Distribution of yellow fever vectors in Northwestern and Western Provinces, Zambia

OBJECTIVE To determine the distribution of yellow fever (YF) vectors species in Northwestern and Western of Zambia, which sampled mosquitoes inside and outside houses in rural, urban, peri-urban and forest areas. METHODS Back-pack aspirators spray catches and CDC light traps collected adult mosquitoes including 405 Aedes, 518 Anopheles, 471 Culex and 71 Mansonia. Morphological vector identification and PCR viral determination were done at a WHO Regional Reference Centre (Institute Pasteur Dakar), Senegal. RESULTS The two main YF vectors were Aedes (Stegomyia) aegypti (Ae. aegypti) and Aedes (Stegomyia) africanus. The first was collected in peri-urban areas and the later was in forest areas, both sparsely distributed in Northwestern Province, where the 0.43 Breteau and 1.92 container indexes, respectively implied low risk to YF. Aedes (Aedimorphus) mutilus; Aedes (Aedimorphus) minutus and Aedes (Finlaya) wellmani were also found in Northwestern, not in Western Province. No Aedes were collected from rural peri-domestic areas. Significantly more Aedes species (90.7%, n=398) than Anopheles (9.1%, n=40) were collected in forest areas (P<0.001) or Culex species (0.2%, n=2) (P<0.001). Ae. aegypti was found only in a discarded container but not in flower pots, old tyres, plant axils, discarded shallow wells, disused container bottles and canoes inspected. CONCLUSIONS Ae. aegypti and Aedes africanus YF vectors were found in the study sites in the Northwestern Province of Zambia, where densities were low and distribution was sparse. The low Breteau index suggests low risk of YF in the Northwestern Province. The presence of Aedes in Northwestern Province and its absence in the Western Province could be due to differing ecological factors in the sampled areas. Universal coverage of vector control interventions could help to reduce YF vector population and the risk to arthropod-borne virus infections.

Individual and Household Level Risk Factors Associated with Malaria in Nchelenge District, a Region with Perennial Transmission: A Serial Cross-Sectional Study from 2012 to 2015

Background The scale-up of malaria control interventions has resulted in substantial declines in transmission in some but not all regions of sub-Saharan Africa. Understanding factors associated with persistent malaria transmission despite control efforts may guide targeted interventions to high-risk areas and populations. Methods Household malaria surveys were conducted in Nchelenge District, Luapula Province, in northern Zambia. Structures that appeared to be households were enumerated from a high-resolution satellite image and randomly sampled for enrollment. Households were enrolled into cross-sectional (single visit) or longitudinal (visits every other month) cohorts but analyses were restricted to cross-sectional visits and the first visit to longitudinal households. During study visits, a questionnaire was administered to adults and caretakers of children and a blood sample was collected for a malaria rapid diagnostic test (RDT) from all household residents. Characteristics associated with RDT positivity were analyzed using multi-level models. Results A total of 2,486 individuals residing within 742 households were enrolled between April 2012 and July 2015. Over this period, 51% of participants were RDT positive. Forty-three percent of all RDT positive individuals were between the ages of 5 and 17 years although this age group comprised only 30% of study participants. In a multivariable model, the odds being RDT positive were highest in 5–17 year olds and did not vary by season. Children 5–17 years of age had 8.83 higher odds of being RDT positive compared with those >18 years of age (95% CI: 6.13, 12.71); there was an interaction between age and report of symptoms, with an almost 50% increased odds of report of symptoms with decreasing age category (OR = 1.49; 95% CI 1.11, 2.00). Conclusions Children and adolescents between the ages of 5 and 17 were at the highest risk of malaria infection throughout the year. School-based programs may be effective at targeting this high-risk group.

Complete Anopheles funestus mitogenomes reveal an ancient history of mitochondrial lineages and their distribution in southern and central Africa

Anopheles funestus s.s. is a primary vector of malaria in sub-Saharan Africa. Despite its important role in human Plasmodium transmission, evolutionary history, genetic diversity, and population structure of An. funestus in southern and central Africa remains understudied. We deep sequenced, assembled, and annotated the complete mitochondrial genome of An. funestus s.s. for the first time, providing a foundation for further genetic research of this important malaria vector species. We further analyzed the complete mitochondrial genomes of 43 An. funestus s.s. from three sites in Zambia, Democratic Republic of the Congo, and Tanzania. From these 43 mitogenomes we identified 41 unique haplotypes that comprised 567 polymorphic sites. Bayesian phylogenetic reconstruction confirmed the co-existence of two highly divergent An. funestus maternal lineages, herein defined as lineages I and II, in Zambia and Tanzania. The estimated coalescence time of these two mitochondrial lineages is ~500,000 years ago (95% HPD 426,000–594,000 years ago) with subsequent independent diversification. Haplotype network and phylogenetic analysis revealed two major clusters within lineage I, and genetic relatedness of samples with deep branching in lineage II. At this time, data suggest that the lineages are partially sympatric. This study illustrates that accurate retrieval of full mitogenomes of Anopheles vectors enables fine-resolution studies of intraspecies genetic relationships, population differentiation, and demographic history. Further investigations on whether An. funestus mitochondrial lineages represent biologically meaningful populations and their potential implications for malaria vector control are warranted.

Risk Factors for Mortality in Children Hospitalized with Severe Malaria in Northern Zambia: A Retrospective Case-Control Study

Abstract. Malaria remains a public health crisis in areas where it has resisted control efforts. In Nchelenge District, a high-transmission area in northern Zambia, malaria accounts for more than one-third of pediatric hospitalizations and nearly one-half of hospital deaths in children. To identify risk factors for death due to malaria, we conducted a retrospective, time-matched case-control study of 126 children hospitalized with malaria who died (cases) and 126 children who survived (controls). There were no differences in age, gender, hemoglobin concentration, or prevalence of severe anemia between cases and controls. Children who died were more likely to come from villages located at greater distances from the hospital than children who survived (median 13.5 versus 3.2 km). Each additional kilometer of distance from the hospital increased the odds of death by 4% (odds ratio 1.04, 95% confidence interval 1.01–1.07, P < 0.01). Extent of anemia and admission during periods when blood was unavailable for transfusion were associated with early death (P ≤ 0.03). Delays in initiation of treatment of severe malaria contribute to the increased odds of death in children referred from more distant health centers, and might be mitigated by transportation improvements, capacity at rural health posts to administer treatment before transfer, hospital triage systems that minimize time to treatment, and reliable blood product stores at referral hospitals.

SEASONAL ABUNDANCE AND SPOROZOITE RATES IN MALARIA VECTORS IN NCHELENGE INCLUDING ISLANDS OF LAKE MWERU AN AREA WITH A HIGH BURDEN OF MALARIA IN NORTHERN ZAMBIA

Background Nchelenge district is a holoendemic malaria transmission zone in northern Zambia. The district occurs in a region characterised by a mix of water, marshes, islands and lagoons presenting a uniquely suitable ecology for mosquitoes. Annual indoor residual spraying (IRS) campaigns are carried out between September and December synchronised with other regions in the country with different environmental conditions. Targeted vector control interventions have been applied since 2008 without appreciable impact on disease burden. The timing and targeting of vector control measures are not guided by an informed entomological baseline. This study was aimed at providing entomological information on the seasonal abundance, spatial distribution and Plasmodium falciparum sporozoite infections in the local malaria vector species in order to guide implementation of vector control in the district. Methods Entomological studies were conducted intermittently spanning the rainy, cold-dry, and hot-dry seasons from 2015–2016. Mosquitoes were collected by CDC light traps, identified to species both morphologically and by PCR techniques. Circumsporozoite ELISA assay was used to detect P. falciparum in mosquito salivary glands. Results A total of 5437 female Anopheles funestus and An. gambiae and over 6000 culicines, mostly Mansonia mosquitoes were collected. The peak number of the An. funestus from all sites occurred in July. Overall P. falciparum infection rates in An. funestus were Kilwa island 2.7% (4/146), Mainland 2.5% (3/122), Chisenga island 0.4% (1/220), Isokwe 5.9% (2/34) and in An. leesoni from Kilwa 33% (1/3). The highest number of An. gambiae was collected from Kilwa and none was found infected with P. falciparum regardless of collection site. Conclusions The annual IRS conducted between September and December may be ineffective in controlling malaria as this misses the vector peak abundance and peak transmission season. Two rounds of IRS covering more areas would be needed to control the two vector species with different population peak seasons and malaria transmission.