Biology subjects

Govella, N. J.

Publications and source records attributed to Govella, N. J..

11 recordsLinked to original sources

bioRxiv2026

Exploring the diversity and spatial distribution of Anopheles mosquitoes and their associated Plasmodium species in Tanzania using the ANOSPP amplicon panel

Species-level identification of malaria vectors and parasites is essential for targeted vector control, surveillance, and clinical case management. However, routine surveillance often relies on morphology, species group-restricted PCR assays, and parasite-detection workflows optimised for Plasmodium falciparum, potentially masking wider vector and parasite diversity. To address this gap, we applied the previously published ANOSPP amplicon sequencing panel to Anopheles mosquitoes collected across 25 districts in mainland Tanzania between 2020 and 2023, using it to characterise vector and parasite diversity, evaluate routine species assignments, and examine how primary-vector occurrence related to national malaria endemicity patterns. Of 71,146 mosquitoes collected, 10,020 were morphologically identified as Anopheles. ANOSPP-based identification resolved 15 Anopheles taxa, including the first Tanzanian record of Anopheles funestus-like mosquitoes (n = 10) and the first resolution of Anopheles longipalpis to Type C in the country (n = 13). Using ANOSPP as the reference, morphology misclassified 6.2% of ANOSPP-analysed specimens, with Anopheles rufipes and Anopheles maculipalpis frequently misclassified, and Anopheles pretoriensis and Anopheles marshallii sensu lato completely missed. Five Plasmodium species were detected across 37 parasite-positive mosquitoes: P. falciparum, P. vivax, P. ovale, P. malariae, and P. caprae. Plasmodium caprae was detected only in Anopheles arabiensis (n = 6), representing the first record of this parasite in Tanzania and its first detection in An. arabiensis globally. Co-occurrence of An. arabiensis, An. funestus s.s., and An. gambiae s.s. was more common in higher malaria transmission strata, whereas occurrence of An. arabiensis alone predominated in very-low and low strata. These findings show that integrating ANOSPP into surveillance can substantially improve vector and parasite resolution, uncover overlooked diversity, and strengthen species-informed malaria surveillance. Author summaryEffective malaria control requires accurate data on mosquito species distributions and the parasites they carry. However, in many settings surveillance still relies mainly on morphological identification or molecular tests targeting limited vector groups, while parasite detection often focuses on Plasmodium falciparum. This limits understanding of mosquito and parasite diversity, obscuring transmission dynamics. We used the previously published ANOSPP amplicon panel to analyse Anopheles mosquitoes across Tanzania. This revealed hidden biological complexity with direct implications for malaria transmission, resolving 15 mosquito and five Plasmodium species. Notably, for the first time in Tanzania, we detected the goat-associated parasite Plasmodium caprae in Anopheles arabiensis and identified An. funestus-like, whose vectorial capacity remains unclear, and An. longipalpis Type C, implicated in malaria transmission in dryland Kenya. Interestingly, co-occurrence of all three major Tanzanian vectors aligned with higher transmission settings, whereas areas where An. arabiensis was the sole vector were mainly in lower transmission settings. These findings show that species-resolved surveillance can reveal hidden biological complexity and provide a stronger basis for locally targeted malaria control.

genomics↗

bioRxiv2026

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↗

bioRxiv2025

Microsatellite Analysis of Genetic Differentiation Among Populations of the Malaria Vector, Anopheles funestus, across Mainland Tanzania Reveals Contrasted Patterns of Geographic Isolation and Gene Flow

BackgroundDespite Anopheles funestus s.s. being a highly competent and widespread malaria vector in Africa, its population structure remains largely understudied in many countries, including Tanzania. Herein, we examine the genetic diversity, geographic isolation, and gene flow of An. funestus populations across 10 administrative regions in mainland Tanzania. MethodsWe employed 12 previously used microsatellite DNA markers to describe genetic diversity, isolation by distance, and gene flow patterns among 10 An. funestus s.s. populations (n = 654) and one An. parensis population (n = 28), used as an outgroup, sampled across 10 regions in mainland Tanzania. ResultsOverall, allelic richness (Na) and genetic diversity (HS) did not differ significantly among populations. Although some loci and populations showed significant departures from Hardy-Weinberg equilibrium, the patterns were not indicative of sub-structuring within locations. Pairwise genetic divergence (FST) values indicated clear separation between An. parensis and An. funestus s.s., with values exceeding 0.2, consistent with species-level differentiation. Among An. funestus s.s. populations, the highest divergence was observed between southeastern coastal populations (Mtwara, Ruvuma, Lindi) and inland populations, with FST values up to 0.288. There was no evidence of isolation by distance. Instead, patterns of genetic divergence suggested connectivity across the Rift Valley and heterogeneity among southeastern populations. Neighbour-joining analysis and Bayesian genotype clustering identified three distinct population groups: (i) An. parensis (Dodoma), (ii) a genetically distinct An. funestus s.s. population from Mtwara, and (iii) a more homogeneous cluster comprising the remaining An. funestus s.s. populations. Notably, the Mtwara population appeared highly differentiated, with divergence approaching that between An. Funestus s.s. and An. parensis, supporting its distinctiveness but not undermining the role of An. parensis as an outgroup. ConclusionExcept for the Mtwara population, whose status will need to be clarified through whole genome sequencing, moderate genetic divergence was found among An. funestus s.s. populations across Tanzania, despite geographical separation and the Rift Valley. The observed genetic structure suggests that anthropogenic gene flow may play a key role in shaping population divergence. Future studies should aim to delineate the effects of local adaptation from recent gene flow to further explore these dynamics.

genetics↗

bioRxiv2025

Characterization of a new laboratory colony of Anopheles funestus mosquitoes established in Ifakara, Tanzania

BackgroundAnopheles funestus, a major vector of malaria in Africa, has proven difficult to colonize in laboratory settings, impeding research on its biology and control. After several attempts, our team recently succeeded in colonizing a strain of An. funestus from Tanzania (FUTAZ). The objective of this study was to analyse the key fitness and genotypic characteristics of these mosquitoes during multiple filial generations of laboratory adaptation and compare them to wild An. funestus from Tanzania and a pre-existing colony of An. funestus from Mozambique (FUMOZ). MethodsMeasures of mating success (percentage of female mosquitoes inseminated), body size (wing length), fecundity (number of eggs laid per female), and insecticide susceptibility (percentage of 24-hour mortality after exposure to insecticides) were compared between the newly established colonies of Tanzanian An. funestus (FUTAZ colonies), the long-established FUMOZ colonies, and a colony of Anopheles arabiensis maintained in the same laboratory. The maternal lineages of the An. funestus mosquitoes were investigated through a hydrolysis probe analysis of their mitochondrial DNA to identify distinct clades, I and II. Additionally, other intragenomic variations were examined through a PCR analysis of restriction fragment length polymorphisms (RFLP) on the third domain of 28S ribosomal DNA. These molecular markers were used to compare the FUTAZ colonies, FUMOZ colonies in Tanzania and South Africa, and the wild-collected An. funestus from Tanzania. ResultThe mating success and body size of FUTAZ females declined significantly from filial generations F1 to F6 relative to the founder population (F0), but then increased from F7 onwards eventually matching FUMOZ by F9. Fecundity was similar across all colonies tested. However, it took significantly longer for 50% of the females in the FUTAZ and FUMOZ colonies (over 10 days) to mate compared to females in the An. arabiensis colony (approximately 5 days). Insecticide resistance appeared to be lost during colonization, but this varied with insecticide classes. Majority of mosquitoes in the FUTAZ colony, as well as the wild-caught Tanzanian An. funestus belonged to Clade I (80.4-89.4%) and RFLP type "Y" (90.5-91.4%), while the FUMOZ colonies were mostly Clade II (65.5-88.5%) and RFLP type "MW" (90.5-91.5%). ConclusionThis study suggests that the mating success and body size of An. funestus decreases significantly during the early stages of colonization, then increase as the mosquitoes adapt to laboratory conditions. It is therefore crucial to have a large enough founder population to persist through these early generations in order to achieve stable colonization of An. funestus. The Clade and RFLP genotyping demonstrated the genetic similarities between the FUTAZ mosquitoes and wild-caught Tanzanian An. funestus, but also showed that the new colony can be distinguished from the FUMOZ colony.

ecology↗

bioRxiv2025

Distinct Genetic Populations and Resistance Backgrounds of the Malaria Vector Anopheles funestus in Tanzania

Population genetic analysis of mosquitoes is becoming increasingly important for understanding the distribution of insecticide resistance alleles, devising sustainable insecticide-based vector control approaches, and how malaria vector populations are structured in space. Anopheles funestus is the dominant malaria vector in Tanzania and most parts of East and Southern Africa. To better understand its population genomic structure in Tanzania, we sequenced the genomes of 334 individual An. funestus mosquitoes from 11 administrative regions. We found two genetically differentiated populations; one inland and at high altitude (found in Katavi, Kagera, Kigoma, and Mwanza) and a second coastal, at low altitude (found in Pwani, Morogoro, Tanga, Ruvuma, Mtwara, Dodoma, and Lindi), with differences in genetic diversity and inbreeding. We found asynchronous selective sweeps, associated with insecticide resistance phenotypes, at the Cyp9k1 gene, and Cyp6p gene cluster, with distinct copy number-variant profiles between the coastal and inland populations. These results suggest that inland and coastal An. funestus populations have divergent histories, with the arid, central region of Tanzania, which also contains the Rift Valley being a possible barrier to gene flow. Such population disconnectedness should be considered for insecticide deployment, resistance management, and the rollout of novel genetic- based vector control approaches. These findings provide the most detailed study of Tanzanian An. funestus population structure and resistance genetics to date. Future research should examine the epidemiological relevance of this discontinuity in gene flow and whether these populations have different malaria transmission abilities.

genetics↗

bioRxiv2024

Community-Based Entomological Surveillance and Control of Vector-Borne Diseases: A Scoping Review

Community-based surveillance and control methods (CBMs) present opportunities to decentralize surveillance and control efforts while simultaneously enhancing community education, leadership, and participation in the fight against vector-borne diseases (VBDs). A scoping review was conducted to describe how CBMs are being utilized currently to combat malaria, dengue fever, Chagas disease, tick-borne diseases (TBDs) and other mosquito-borne diseases (MBD) exclusive of dengue and malaria, and to overall highlight key approaches, lessons learned, potential challenges, and recommendations. A total of 304 potential publications were identified among which 82 met the inclusion criteria. This scoping review highlighted the following benefits to CBMs: cost savings, increased sustainability, increased community knowledge, human behavior changes, increased surveillance coverage, ease in deployment, and the creation of larger, more diverse entomological datasets. Potential challenges highlighted include: participant retention and motivation, participant recruitment and incentives, continued governmental support, data quality, and collaboration with local municipal authorities. CBMs are commonly and successfully used in vector surveillance and control systems, but the chosen vector management method varies by vector-borne disease and region of the world. Additional research is needed to support the implementation of CBMs including cost-effectiveness studies and those studies with negative outcomes. Taken together, this scoping review highlights key aspects, potential challenges, and benefits of CBMs, and outlines potential future directions for incorporating CBMs into VBD control and elimination programming, and potential for community based integrated vector management (IVM) approaches.

scientific communication and education↗

bioRxiv2024

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↗

bioRxiv2024

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↗

bioRxiv2024

Discovery of knock-down resistance in the major malaria vector Anopheles funestus reveals the legacy of persistent DDT pollution.

A major mechanism of insecticide resistance in insect pests is knock-down resistance (kdr) caused by mutations in the voltage-gated sodium channel (Vgsc) gene. Despite being common in most malaria Anopheles vector species, kdr mutations have never been observed in Anopheles funestus, the principal malaria vector in Eastern and Southern Africa. While monitoring 10 populations of An. funestus in Tanzania, we unexpectedly found resistance to DDT, a banned insecticide, in one location. Through whole-genome sequencing of 333 An. funestus samples from these populations, we found 8 novel amino acid substitutions in the Vgsc gene, including the kdr variant, L976F (L1014F in An. gambiae), in tight linkage disequilibrium with another (P1842S). The mutants were found only at high frequency in one region, with a significant decline between 2017 and 2023. Notably, kdr L976F was strongly associated with survivorship to the exposure to DDT insecticide, while no clear association was noted with a pyrethroid insecticide (deltamethrin). Further study is necessary to identify the origin and spread of kdr in An. funestus, and the potential threat to current insecticide-based vector control in Africa. SignificanceKnock-down resistance (kdr) mutations confer resistance to malaria vector control insecticides and pose a grave threat to malaria control. Here, we report the first discovery of kdr in An. funestus, the principal malaria vector in East and Southern Africa. Kdr in An. funestus conferred resistance to DDT but not deltamethrin. Based on extensive DDT contamination and unofficial usage in Tanzania, it is possible that kdr emerged because of widespread organic pollution as opposed to through public health efforts. Regardless of origin, the discovery of kdr in An. funestus is an alarming development that warrants immediate, extensive follow-up and close surveillance to establish the origin, and extent to which it may threaten malaria control in An. funestus.

genetics↗

bioRxiv2021

Heritability and phenotypic plasticity of biting time behaviors in the major African malaria vector Anopheles arabiensis

Use of Insecticide Treated Nets for malaria control has been associated with shifts in mosquito vector feeding behavior including earlier and outdoor biting on humans. The relative contribution of phenotypic plasticity and heritability to these behavioural shifts is unknown. Elucidation of the mechanisms behind these shifts is crucial for anticipating impacts on vector control. We used a novel portable semi-field system (PSFS) to experimentally measure heritability of biting time in the malaria vector Anopheles arabiensis in Tanzania. In PSFS assays, the biting time of F2 offspring (early: 18:00-21:00, mid: 22:00-04:00 or late: 05:00-07:00) was significantly associated with that of their wild-caught F0 grandmothers, corresponding to an estimated heritability of 0.30 (95% CI: 0.20, 0.41). F2 from early-biting F0 were more likely to bite early than F2 from mid or late late-biting F0. Similarly, the probability of biting late was higher in F2 derived from mid and late-biting F0 than from early-biting F0. Our results indicate that variation in biting time is attributable to additive genetic variation. Selection can therefore act efficiently on mosquito biting times, highlighting the need for control methods that target early and outdoor biting mosquitoes.

evolutionary biology↗

bioRxiv2020

An Autoencoder and Artificial Neural Network-based Method to Estimate Parity Status of Wild Mosquitoes from Near-infrared Spectra

BackgroundAfter mating, female mosquitoes need animal blood to develop their eggs. In the process of acquiring blood, they may acquire pathogens, which may cause different diseases to humans such as malaria, zika, dengue, and chikungunya. Therefore, knowing the parity status of mosquitoes is useful in control and evaluation of infectious diseases transmitted by mosquitoes, where parous mosquitoes are assumed to be potentially infectious. Ovary dissections, which currently are used to determine the parity status of mosquitoes, are very tedious and limited to very few experts. An alternative to ovary dissections is near-infrared spectroscopy (NIRS), which can estimate the age in days and the infectious state of laboratory and semi-field reared mosquitoes with accuracies between 80 and 99%. No study has tested the accuracy of NIRS for estimating the parity status of wild mosquitoes. Methods and resultsIn this study, we train artificial neural network (ANN) models on NIR spectra to estimate the parity status of wild mosquitoes. We use four different datasets: An. arabiensis collected from Minepa, Tanzania (Minepa-ARA); An. gambiae collected from Muleba, Tanzania (Muleba-GA); An. gambiae collected from Burkina Faso (Burkina-GA); and An.gambiae from Muleba and Burkina Faso combined (Muleba-Burkina-GA). We train ANN models on datasets with spectra preprocessed according to previous protocols. We then use autoencoders to reduce the spectra feature dimensions from 1851 to 10 and re-train ANN models. Before the autoencoder was applied, ANN models estimated parity status of mosquitoes in Minepa-ARA, Muleba-GA, Burkina-GA and Muleba-Burkina-GA with out-of-sample accuracies of 81.9 {+/-} 2.8% (N=927), 68.7 {+/-} 4.8% (N=140), 80.3 {+/-} 2.0% (N=158), and 75.7 {+/-} 2.5% (N=298), respectively. With the autoencoder, ANN models tested on out-of-sample data achieved 97.1 {+/-} 2.2%, (N=927), 89.8 {+/-} 1.7% (N=140), 93.3 {+/-} 1.2% (N=158), and 92.7 {+/-} 1.8% (N=298) accuracies for Minepa-ARA, Muleba-GA, Burkina-GA, and Muleba-Burkina-GA, respectively. ConclusionThese results show that a combination of an autoencoder and an ANN trained on NIR spectra to estimate parity status of wild mosquitoes yields models that can be used as an alternative tool to estimate parity status of wild mosquitoes, especially since NIRS is a high-throughput, reagent-free, and simple-to-use technique compared to ovary dissections.

bioinformatics↗