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Msoffe, R. V.

Publications and source records attributed to Msoffe, R. V..

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An improved generic schema for high fidelity data linkage and sample tracing across complex multi-assay medical entomology studies

Evidence-based decision making on malaria vector control strategies increasingly rely on triangulation of data which requires informatics systems that can integrate data from complex, multi-stage studies involving mosquitoes. This manuscript describes a performance evaluation of an extended version of the generic schema underpinning the VBDs360 platform, specifically improved to accommodate multiple distinct entomological assays spanning the field, insectary and laboratory. The utility of this extension, with respect to high-fidelity data linkage and robust sample traceability across complex entomological workflows, was evaluated through a case study conducted in southern Tanzania. Wild female mosquitoes were collected from 40 locations across a >4,000 km{superscript 2} area and then reared through multiple generations in an insectary before derived iso-female lineages were tested for phenotypic susceptibility to a pyrethroid insecticide. Such multi-generational lineages (F to F where n [≥] 2) were propagated to prevent non-heritable maternal effects on phenotype and produce enough progeny for standard WHO susceptibility assays. All samples were subsequently archived in a molecular laboratory, where all F specimens were tested for sibling species identity. A paper-based implementation of the extended schema enabled successful integration of 77,017 lines of data distributed across 6 different tables that spanned 3 distinct field, insectary, and laboratory workflows, implemented by three different teams working in different locations. At each step, fully independent and redundant primary and secondary keys enabled high fidelity error correction and sample tracing. Consistently perfect linkage between assay design and sample sorting data was achieved for F0 wild-caught adults, with 100% of 66,108 record successfully linked between field capture and morphological categorization. This complete traceability extended to the propagation of derived Fn lineages, with all 100 and 243 records from 9 adult-derived and 13 larval-derived lineages, respectively, correctly linked. Insecticide susceptibility phenotype further confirmed 100% linkage for 5,654 records between exposure history and recorded mortality outcome data in the insectary. Although such cross-cleaned linkages to sample analysis and storage data recorded by the laboratory team were not entirely perfect and could be improved, they were nevertheless of very high fidelity (97.3% (1967/2,022) for F0 samples and 99.3% (437/440) for Fn samples). Overall, this pilot application of the extended generic schema ensured robust data provenance and minimized transcription errors in this complex study distributed across multiple teams and locations. These findings demonstrate how this generic informatics framework may be scaled and adapted to support data integrity across diverse, large-scale, multi-team entomological research workflows.

bioinformatics↗

Persistence of residual Anopheles gambiae populations associated with frontier human settlements and nocturnal livelihood-related activities at the fringes of a large conservation area in southern Tanzania

Following the rapid scale-up of long-lasting insecticidal nets (LLINs) in 2008 across the Kilombero Valley of southern Tanzania, Anopheles gambiae Giles, a highly efficient and human-specialized malaria vector, essentially disappeared within two years and has rarely been detected since. However, an ecological study of its sibling species within the An. gambiae complex, namely An. arabiensis Patton and An. quadriannulatus Theobald, in an area spanning human settlements and conserved wilderness, nevertheless detected sparse, highly focal populations of An. gambiae. Out of 4,704 An. gambiae complex specimens collected, only 10 were identified as nominate An. gambiae by polymerase chain reaction. Seven of these were captured at three surveyed fishing camps inside a Wildlife Management Area (WMA), while none were caught in five established villages west of the WMA or the national park to the east. Two of the other three were found among scattered farming settlements along the edge of the WMA. The final individual was caught in a near-natural area deep inside the WMA, but even this site was near small transient homesteads. Poisson regression modelling confirmed that capture rates at fishing camps were higher than in the peripheral settlements (Relative Rate [RR] [95% Confidence Interval (CI)] = 0.088 [0.011, 0.695], P = 0.021) or conserved natural areas (RR [95% CI] = 0.043 [0.003, 0.495], P = 0.012). Conservative estimates put the mean standing populations at each of the three fishing camps between 26 and 41 females per settlement. The absence of An. gambiae from well-conserved areas without resident humans aligns with its anthropophagic nature and dependency on human hosts. Residual An. gambiae populations persisting among frontier communities appear linked to locally common, livelihood-related nocturnal activities that preclude the practical use of LLINs, specifically fishing and guarding crops against wildlife. The continued presence of this previously important species in these ecological niches, where local livelihoods apparently undermine high LLIN coverage, shows persistence under sustained, partial insecticide pressure. This could lead to the re-emergence of this vector through the gradual evolution of resistance. However, their sparse and focal distribution suggests that new supplementary interventions, such as transfluthrin emanators, might effectively eliminate these remaining refuge populations.

ecology↗

Comparative attractiveness of Anopheles quadriannulatus and Anopheles arabiensis to humans estimated by comparing the relative abundance of these two species in larval samples, unbaited adult catches and human-baited adult catches.

Understanding mosquito vector behaviour, abundance, and bionomics is crucial for effective malaria prevention. Since most malaria parasites in humans are strict anthroponoses, mosquito preference for humans as a blood source is a key determinant of transmission intensity and intervention strategies. This study compares the attraction of Anopheles arabiensis and Anopheles quadriannulatus to humans by assessing their relative abundance in larval samples and adult mosquito catches using unbaited and human-baited traps. The research investigated how the abundance of these sibling species varies with the local availability of humans, livestock, and wildlife as potential blood sources. Surveys of larval and adult mosquito populations were conducted at 40 mobile camping locations distributed across a landscape mosaic of different habitat types, with a gradient of land use practices ranging from comprehensive conversion to agriculture and human settlement to essentially intact natural ecosystems inside well-protected conservation areas. Larvae were collected from all accessible water bodies within a 2 km radius of each mobile camp, while adults were surveyed using four light traps and one interception netting barrier trap at each camp. Light traps were strategically placed at locations such as beside a human-occupied tent, near the camp, in a nearby streambed, and in an open natural glade, while the netting barrier trap was placed in the open natural glade. Most adult Anopheles gambiae complex mosquitoes caught were unfed and presumably host-seeking. The mean catches of the complex in light traps next to the human-occupied tent was over four times higher than elsewhere around the camp (p<<0.0001). Catches decreased with distance from humans, suggesting attraction to humans by at least one species in the complex. An. arabiensis was caught in greater numbers in the human-occupied tent, while An. quadriannulatus catches remained consistently low across all traps, even in wild areas where it was dominant in larval population. The proportion of An. arabiensis in adult collections was higher than in larval samples (98.7% versus. 78.3%, p<<0.0001), and adults caught beside human-occupied tents had 36 times higher odds of being An. arabiensis rather than An. quadriannulatus. Similarly, the barrier trap away from humans but frequently visited by researchers showed a 24-fold enrichment of An. arabiensis. These results confirm the strong attraction of An. arabiensis to humans, contrasting with the non-vector An. quadriannulatus, which is largely unresponsive to humans. Light traps by human-occupied tents efficiently capture anthropophagic mosquitoes outdoors, while unbaited traps far from people appear to give unbiased representations of larval population composition but with very low efficiency. Moreover, netting barriers with human activity attract anthropophagic mosquitoes, turning them into semi-baited traps with moderate efficiency.

ecology↗

Blood host preferences and competitive inter-species dynamics within an African malaria vector species complex inferred from signs of animal activity around aquatic larval habitats.

The dual-specialized behavioural adaptions of Anopheles arabiensis, to feed readily upon either people or cattle, enable it to thrive and mediate persistent residual malaria transmission across much of Africa, despite widespread use of long-lasting insecticidal nets (LLINs). Indeed, LLIN scale up has often resulted in it dominating the more efficient but vulnerable malaria vector Anopheles gambiae, its sibling species within a complex of the latter name. However, the feeding behaviours and competitive relationships of An. arabiensis with other sibling species in well conserved natural ecosystems, where its known preferred hosts are scarce or absent, remain largely unexplored. Potential aquatic habitats were surveyed for An. gambiae complex larvae across a gradient of natural ecosystem integrity in southern Tanzania, encompassing fully domesticated human settlements, a partially encroached Wildlife Management Area (WMA), and well conserved areas of the recently gazetted Nyerere National Park (NNP) before substantive development of tourist access or accommodation. Direct observations, tracks, spoor and other signs of human, livestock or wild animal activity around these water bodies were recorded as indirect indicators of potential blood source availability. While only An. arabiensis was found in fully domesticated ecosystems, its non-vector sibling species An. quadriannulatus occurred in conserved areas and dominated the most intact natural ecosystems. Proportions of larvae identified as An. arabiensis were positively associated with human and/or cattle activity and negatively associated with distance inside NNP and away from human settlements. Proportions of An. quadriannulatus were positively associated with activities of impala and bushpig, implicating both as likely preferred blood hosts. While abundant impala and lack of humans or cattle in intact acacia savannah within NNP apparently allowed it to dominate An. arabiensis, presence of bushpig seemed to provide it with a foothold in miombo woodlands of the WMA, despite encroachment by people and livestock. While this antelope and suid are essentially unrelated, both are non- migratory residents of small home ranges with perennial surface water, representing preferred hosts for An. quadriannulatus that are widespread across extensive natural ecosystems all year round. Despite dominance of An. quadriannulatus in well-conserved areas, An. arabiensis was even found in absolutely intact environments >40km inside NNP, suggesting it can survive on blood from one or more unidentified wild species. While self-sustaining refuge populations of An. arabiensis inside conservation areas, supported by wild blood hosts that are fundamentally beyond the reach of insecticidal interventions targeted at humans or their livestock, may confound efforts to eliminate this key malaria vector, they might also enable insecticide resistance management strategies that could restore the effectiveness of pyrethroids in particular.

ecology↗

A self-cooling self-humidifying mosquito carrier backpack for transporting live adult mosquitoes on foot over long distances under challenging field conditions.

It is often necessary to use motorized transport to move live mosquitoes from distant field collection points into a central insectary, so that their behavioural and/or physiological phenotypes can be assessed under carefully controlled conditions. However, a survey of heritable insecticide susceptibility traits among wild-caught Anopheles arabiensis mosquitoes, collected across an extensive study area composed largely of wilderness in southern Tanzania, necessitated that live mosquitoes were carried on foot over distances up to 25 km per day because most of the area was impassable by car, motorcycle or even bicycle during the rains. A self-cooling, self-humidifying carrier backpack was therefore developed that allows live adult mosquito specimens to be transported across rugged miombo woodland and floodplain terrain throughout the year. This wettable backpack was fabricated from stitched Tanzanian kitenge cotton fabric and PVC-coated fiberglass netting that allows easy circulation of air in and out. An outer cover flap made of cotton towelling embedded inside a kitenge envelope overhangs the fiberglass netting upper body of the bag, to protect mosquitoes from direct sunlight, and can be soaked with water to maintain low temperature and high humidity inside. Mean survival of insectary-reared female Anopheles arabiensis transported through 9 different mobile camps inside the 509 km2 Ifakara-Lupiro-Mangula Wildlife Management Area (ILUMA WMA), over up to 143km and 25 days, was indistinguishable from those left in the field insectary over the same period. Although considerable variance was observed between different batches of mosquitoes from the insectary and between individual cups of mosquitoes, the different levels and positions inside the backpack had no influence on survival. Temperature and humidity inside the backpack were maintained at standard insectary condition throughout, despite much more extreme conditions immediately outside. When the backpack was applied to collecting wild An. arabiensis and Anopheles quadriannulatus across a much larger study area of >4000 km2, encompassing the ILUMA WMA, some nearby villages and adjacent parts of Nyerere National Park (NNP), it achieved a mean survival rate of 58.2% [95% confidence interval 47.5 to 68.2]. Encouragingly, no difference in survival was observed between ILUMA WMA and NNP even though transport back from NNP involves much longer distances, sometimes involving lengthy journeys by car or boat. Overall, this mosquito carrier backpack prototype appears to represent a viable and effective method for transporting live wild-caught mosquitoes on foot across otherwise impassable terrain under challenging weather conditions with minimal detrimental impact on their survival.

ecology↗