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Esterhuizen, J.

Publications and source records attributed to Esterhuizen, J..

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Evaluation of the vectorial competence of Glossina brevipalpis in the transmission of Trypanosoma congolense-savannah type, in the Matutuine District, Maputo Province, Mozambique

African Animal Trypanosomosis (AAT) is a parasitic disease caused by trypanosome species, posing a major challenge to animal health and production in Africa. In the Matutuine District, southern Mozambique, where Glossina brevipalpis is prevalent, AAT is endemic. Previous research conducted in South Africa suggested that G. brevipalpis plays a marginal role in the transmission of Trypanosoma congolense. However, considering the wide distribution of G. brevipalpis in Southeast Africa and the major epidemiological implications of the aforementioned observations, this study was designed to re-assess the vectorial competence of G. brevipalpis in the transmission of T. congolense-savannah type. The research involved 915 G. brevipalpis flies, fed six times on two cattle infected with T. congolense. After 48 hours, the flies were later fed on a non-infected animal to clean the proboscis, followed by extended feeding on four susceptible cattle for 22 days. These five clean animals were monitored for 60 days based on body temperature, packed cell volume, buffy coat technique and polymerase chain reaction. In the meantime, flies were dissected weekly to observe the development of trypanosomes in the midgut and proboscis. To discard any possible mechanical transmission by G. brevipalpis 48 hours after an infected blood meal, 97 flies from a colony were fed once on two infected cattle and once on a susceptible bovine. The Spearman correlation and Mann-Whitney U tests were used to analyse the data. Results showed that the four susceptible cattle became infected with T. congolense, while the animal used for proboscis cleaning and discard any mechanical transmission remained uninfected. Dissections revealed that 89% of flies were positive for trypanosome through microscopy and 100% through PCR. These findings confirm that G. brevipalpis is a competent biological vector for transmitting T. congolense, challenging earlier assumptions and emphasizing its role in AAT dynamics.

ecology↗

Impact of a national tsetse control programme to eliminate Gambian sleeping sickness in Uganda: a spatio-temporal modelling study.

IntroductionTsetse flies (Glossina) transmit Trypanosoma brucei gambiense which causes gambiense human African trypanosomiasis (gHAT). As part of national efforts to eliminate gHAT as a public health problem, Uganda implemented a large-scale programme of deploying Tiny Targets, which comprise panels of insecticide-treated material which attract and kill tsetse. At its peak, the programme was the largest tsetse control operation in Africa. Here, we quantify the impact of Tiny Targets and environmental changes on the spatial and temporal patterns of tsetse abundance across north-western Uganda. MethodsWe leverage a 100-month longitudinal dataset detailing Glossina fuscipes fuscipes catches from monitoring traps between October 2010 and December 2019 within seven districts in north-western Uganda. We fitted a boosted regression tree model assessing environmental suitability which was used alongside Tiny Target data to fit a spatio-temporal geostatistical model predicting tsetse abundance across our study area ([~]16,000 km2). We used the spatio-temporal model to quantify the impact of Tiny Targets and environmental changes on the distribution of tsetse, alongside metrics of uncertainty. ResultsEnvironmental suitability across the study area remained relatively constant over time, with suitability being driven largely by elevation and distance to rivers. By performing a counterfactual analysis using the fitted spatio-temporal geostatistical model we show that deployment of Tiny Targets across an area of 4000 km2 reduced the overall abundance of tsetse to low levels (median daily catch = 1.1 tsetse/trap, IQR = 0.85-1.28) with no spatial-temporal locations having high (>10 tsetse/trap/day) numbers of tsetse compared to 18% of locations for the counterfactual. ConclusionsIn Uganda, Tiny Targets reduced the abundance of G. f. fuscipes and maintained tsetse populations at low levels. Our model represents the first spatio-temporal model investigating the effects of a national tsetse control programme. The outputs provide important data for informing next steps for vector-control and surveillance. Key questionsO_ST_ABSWhat is already known on this topic?C_ST_ABSSmall panels of insecticide-treated fabric, called Tiny Targets, are used to attract, and kill riverine tsetse, the vectors of T. b. gambiense which causes gambiense human African trypanosomiasis (gHAT). In large-scale (250-2000 km2) trials conducted in five countries, deployment of Tiny Targets reduced the densities of tsetse by between 60 and >90%. What this study addsWe report an analysis of, and data from, a large-scale ([~]4,000km2) national tsetse control programme, implemented in Uganda to eliminate gHAT as a public health problem. We found that Tiny Targets reduced tsetse abundance across the study period (2011-2019) and maintained densities at low (<1 tsetse/trap/day) levels. We produce maps which detail spatial variances in tsetse abundance in response to vector control. How this study might affect research, practice, or policyIn 2022, Uganda received validation from the World Health Organisation (WHO) that it had eliminated gHAT as a public health problem. The large-scale deployment of Tiny Targets contributed to this achievement. Our findings provide evidence that Tiny Targets are an important intervention for other countries aiming to eliminate gHAT.

ecology↗

Insights into Trypanosomiasis Transmission: Age, Infection Rates and Bloodmeal Analysis of Glossina fuscipes fuscipes in N.W. Uganda

BackgroundTsetse flies (Glossina) transmit species of Trypanosoma which cause human African trypanosomiasis (HAT) and animal African trypanosomiasis (AAT). Measuring the infection status of wild-caught tsetse is an important part of operations to control HAT. In north-west Uganda, we conducted field studies over a 15-month period to compare classical, microscope-based, and PCR-based methods of detecting trypanosomes in tsetse. We also quantified the age structure and host preferences of tsetse. MethodsUsing Pyramidal traps placed along the Kochi River, in Koboko district, 12512 G. fuscipes fuscipes were caught. A subset of females ({not}n= 5051) and males (n= 1221) underwent dissection wherein mouthparts, midguts and salivary glands were screened for trypanosomes. Additionally, the age of the females was estimated using ovarian ageing and the trypanosome-positive status of 1931 females and 438 males was investigated by ITS PCR. Further the bloodmeal sources of 131 tsetse were identified using vertebrate cytochrome b PCR. ResultsInfection rates estimated were significantly greater (1.9-9.3 times) using the PCR-based method compared to the classical dissection-based method. Positive rates for T. brucei sl, T. congolense and T. vivax were 1.6% (1.32-2.24), 2.4% (1.83-3.11and 2.0% (1.46-2.63), respectively by PCR. The abundance and age structure of tsetse populations were relatively stable and the slight seasonal four-fold variation in abundance appeared to be correlated with rainfall. Analyses of age structure suggests a low natural daily mortality of 1.75% (1.62-1.88). The majority of bloodmeals were identified as cattle (39%, 30.5-47.8) and human (37% of meals, 28.4-45.6). ConclusionPCR provides a more sensitive and specific method of estimating the infection rates of all pathogenic species of trypanosome circulating in Koboko. The seasonally stable abundance, low mortality rate and high proportion of bloodmeals from humans may explain, in part, why this district has historically been a focus of sleeping sickness. Author summaryTsetse flies (Glossina) transmit African trypanosomiasis in humans ( sleeping sickness) and livestock ( nagana). Identifying trypanosome species and quantifying infection rates in tsetse is important in understanding and controlling trypanosomiasis. Traditionally this relied on microscopic examination of tsetse, however, this method is unable to reliably identify different species of trypanosome. We compared microscopy and PCR, trypanosome-detection methods, using G. fuscipes fuscipes collected from traps deployed in Koboko district, in North-West Uganda. Our results show that the PCR-based method was 1.9-9.3 times more sensitive than the classical approach and able to identify different species of Trypanosoma, including mixed infections. In collecting tsetse we also obtained data on the seasonal abundance, age and diet of the tsetse population. Analyses of these results showed that natural mortality was low (1.75%/day) among adult females and only a slight seasonal variation in abundance and age structure occurred. The most important hosts were cattle and humans, providing 39% and 37% of bloodmeals, respectively. The PCR method provided a sensitive means of identifying trypanosomes to the species level. The longevity and diet of tsetse in Koboko may explain, why this district was a persistent focus of disease prior to the deployment of Tiny Targets to control tsetse.

ecology↗

Impact of vector control on effective population sizes; empirical evidence for a control- based genetic bottleneck in the tsetse fly Glossina fuscipes

We investigated genetic variation at 37 newly-developed microsatellite loci in populations of the tsetse fly Glossina fuscipes fuscipes captured from the upper and lower reaches of a single hydrographical network within an endemic Human African Trypanosomiasis focus. Our primary aim was to assess the impact of vector control using insecticide-treated baits (Tiny Targets) on genetic structure. We initially used STRUCTURE to delineate geographical boundaries of two stable ancestral reference populations without any history of vector control but marked for either vector control ( intervention) or no control ( non-intervention). We then used the ADMIXTURE model to assess genetic divergence in temporal populations collected after vector control implementation. We applied the Linkage Disequilibrium method to explicitly measure spatial and temporal changes in effective population size (Ne). We observed a significant reduction in Ne coincident with vector control, whereas Ne remained stable in the non-intervention area. Our empirical findings show how classical population genetics approaches detected within a short period of time, a significant genetic bottleneck associated with vector control, and opens up the possibility of using routine genomic surveillance. We have also generated a resource of new genetic markers for studies on the population genetics of tsetse at finer-scale resolution. FundingThis work was funded through a Wellcome Trust Masters Fellowship in Public Health and Tropical Medicine awarded to Allan Muhwezi (103268/Z/13/Z).

evolutionary biology↗

Evidence of the absence of Human African Trypanosomiasis in northern Uganda: analyses of cattle, pigs and tsetse flies for the presence of Trypanosoma brucei gambiense

BackgroundLarge-scale control of sleeping sickness has led to a decline in the number of cases of Gambian human African trypanosomiasis (g-HAT) to <2000/year. However, achieving complete and lasting interruption of transmission may be difficult because animals may act as reservoir hosts for T. b. gambiense. Our study aims to update our understanding of T. b. gambiense in local vectors and domestic animals of N.W. Uganda.\n\nMethodsWe collected blood from 2896 cattle and 400 pigs and In addition, 6664 tsetse underwent microscopical examination for the presence of trypanosomes. Trypanosoma species were identified in tsetse from a subsample of 2184 using PCR. Primers specific for T. brucei s.l. and for T. brucei sub-species were used to screen cattle, pig and tsetse samples.\n\nResultsIn total, 39/2,088 (1.9%; 95% CI=1.9-2.5) cattle, 25/400 (6.3%; 95% CI=4.1-9.1) pigs and 40/2,184 (1.8%; 95% CI=1.3-2.5) tsetse, were positive for T. brucei s.l.. Of these samples 24 cattle (61.5%), 15 pig (60%) and 25 tsetse (62.5%) samples had sufficient DNA to be screened using the T. brucei sub-species PCR. Further analysis found no cattle or pigs positive for T. b. gambiense, however, 17/40 of the tsetse samples produced a band suggestive of T. b. gambiense. When three of these 17 PCR products were sequenced the sequences were markedly different to T. b. gambiense, indicating that these flies were not infected with T. b. gambiense.\n\nConclusionThe absence of T. b. gambiense in cattle, pigs and tsetse accords with the low prevalence of g-HAT in the human population. We found no evidence that livestock are acting as reservoir hosts. However, this study highlights the limitations of current methods of detecting and identifying T. b. gambiense which relies on a single copy-gene to discriminate between the different sub-species of T. brucei s.l.\n\nAuthor SummaryThe decline of annual cases of West-African sleeping sickness in Uganda raises the prospect that elimination of the disease is achievable for the country. However, with the decrease in incidence and the likely subsequent change in priorities there is a need to confirm that the disease is truly eliminated. One unanswered question is the role that domestic animals play in maintaining transmission of the disease. The potential of cryptic-animal reservoirs is a serious threat to successful and sustained elimination of the disease. It is with the intent of resolving this question that we have carried out this study whereby we examined 2088 cattle, 400 pigs and 2184 tsetse for Trypanosoma brucei gambiense, the parasite responsible for the disease. Our study found T. brucei s.l. in local cattle, pigs and tsetse flies, with their respective prevalences as follows, 1.9%, 6.3% and 1.8%. Further analysis to establish identity of these positives to the sub-species level found that no cattle, pigs or tsetse were carrying the pathogen responsible for Gambian sleeping sickness. Our work highlights the difficulty of establishing the absence of a disease, especially in an extremely low endemic setting, and the limitations of some of the most commonly used methods.

microbiology↗