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Background Black screen fly round (BFR) is a mobile sampling method for Glossina morsitans. This technique relies on the ability of operator(s) to capture flies landing on the screen with hand nets. In this study, we aimed to evaluate a vehicle-mounted sticky panel trap (VST) that is independent of the operator’s ability to capture flies against BFR, for effective and rapid sampling of G. m. morsitans Westwood and G. m. centralis Machado. We also determined the influence of the VST colour (all-blue, all-black or 1:1 blue-black), orientation and presence of odour attractants on tsetse catch. Methodology/Principal findings Using randomised block design experiments conducted in Zambia, we compared and modelled the number of tsetse flies caught in the treatment arms using negative binomial regression. There were no significant differences in the catch indices of the three colour designs and for in-line or transversely oriented panels for both subspecies (P > 0.05). When baited with butanone and 1-octen-3-ol, VST caught 1.38 (1.11–1.72; P < 0.01) times more G. m. centralis flies than the un-baited trap. Attractants did not significantly increase the VST catch index for G. m. morsitans (P > 0.05). Overall, the VST caught 2.42 (1.91–3.10; P < 0.001) and 2.60 (1.50–3.21; P < 0.001) times more G. m. centralis and G. m. morsitans respectively, than the BFR. The VST and BFR took 10 and 35 min respectively to cover a 1 km transect. Conclusion/Significance The VST is several times more effective for sampling G. m. morsitans and G. m. centralis than the BFR and we recommend its use as an alternative sampling tool., Author summary The fly round is a mobile method used to sample G. m. morsitans and G. m. centralis, important vectors of human and animal African trypanosomiasis. However, its effectiveness is largely dependent on the skill and ability of the operator(s) to catch flies using a hand net. Here, we report the evaluation of an alternative mobile sampling tool, the vehicle-mounted sticky trap (VST) which is independent of operator skill and ability to catch flies. We show that VST is more effective in catching both female and male G. m. morsitans and G. m. centralis compared to the black-screen fly round (BFR). Furthermore, VST covered the same distance of BFR in a much shorter time. This study provides a basis for the use of VST in large scale sampling of G. morsitans to determine its geographical limit, a critical aspect in the planning of vector control strategies and interventions.

Monitoring the effectiveness of tsetse fly control interventions that aim to reduce transmission of African trypanosomiasis requires highly efficient sampling tools that can catch flies at low densities. The sticky small target (StS-target) has previously been shown to be more effective in sampling Glossina fuscipes fuscipes compared to the biconical trap. However, its efficiency in terms of the proportion of flies it catches out of those that visit it has not been reported. Furthermore, there are no reports on whether tsetse samples caught using the StS-target can be used for subsequent processes such as molecular tests. In this study, we evaluated the efficiency of the biconical trap and targets for sampling G. f. fuscipes. All targets were tiny (0.25 × 0.50 m) but varied in their capture system. We used targets with sticky surface (StS-targets) and those with an electrified surface (ES-targets). We also assessed the suitability of flies caught on the StS-target for molecular tests by amplifying DNA of bacterial communities. Randomized block design experiments were undertaken in Mbita area and Manga Island on Lake Victoria of western Kenya. Fly catches of each sampling tool were compared to those of the sampling tool flanked by electric (E) nets and analyzed using a negative binomial regression. The total catch for each sampling tool alone was divided by the total catch of the sampling tool flanked by two E-nets to obtain its efficiency expressed as a percentage. A proportion of flies caught on the StS-target was preserved for molecular tests. Overall, the efficiencies of the biconical trap, ES-target and StS-target were 7.7%, 13.3% and 27.0%, respectively. A higher proportion of females (69 to 79%) than males approached all the sampling tools, but the trap efficiency was greater for male G. f. fuscipes than females. Furthermore, sequencing the 16S rRNA gene from fly samples caught on the StS-target revealed the presence of Spiroplasma. Our results indicate that the SS-target is the most efficient trap to monitor G. f. fuscipes population during interventions, with the biconical trap being the least efficient, and samples collected from StS-targets are suitable for molecular studies.

Introduction A variety of techniques have been used to control tsetse with varying degrees of success. In a study on the population structure of Glossina fuscipes fuscipes that recovered after a previous vector control trial on two Kenyan islands, it was reported that the average fly size on the intervention islands was significantly smaller than on the none intervention islands and also compared to the size before the intervention. The conclusion was that vector control using tiny targets exerted size selection pressure on the population. The study recommended for further studies and suggested that this phenomenon could be among the reasons why targets used as a sole control method have rare reports of successful elimination of tsetse populations. Therefore, in this paper we report on a study of body size of tsetse flies caught in epsilon traps (as a stationary device) and black screen fly rounds (as a mobile trapping device). Materials and methods The study was carried out in eastern Zambia to test the hypothesis that the body size (measured as wing length) of G. m. morsitans males or females, captured by epsilon traps and fly rounds is the same. Results A total of 1442 (489 females and 953 males) wing length measurements of G. m. morsitans were used in the analysis. It was established that tsetse flies caught by epsilon traps are on average larger than those caught by fly rounds. The likelihood of a large female or male fly being caught by traps, relative to a small one, significantly increased by 5.088 times (95% CI: 3.138–8.429) and by 2.563 times (95% CI: 1.584–4.148), respectively, p Conclusion This study showed that epsilon traps capture significantly larger G. m. morsitans than fly rounds do. Therefore, further research is recommended to verify i) whether the predilection of traps to capture larger flies has an effect on the process of tsetse elimination when targets are used e.g. targets may take longer to reach elimination than if the predilection was not there, ii) whether different results can be obtained on ecogeographic distribution of different sizes of the species if fly rounds are used for sampling instead of epsilon traps. The results from such studies could influence the strategies used in future control operations.

Background Small targets comprising panels of blue and insecticide-treated black netting material each 0.25 × 0.25 m have been shown to attract and kill Glossina fuscipes fuscipes Newstead, 1910 (Diptera: Glossinidae) thereby reducing its population density by over 90% in field trials. However, their attractive ability has not been fully exploited for sampling purposes. Therefore, in this study we assessed the effectiveness of using sticky small targets as sampling tools for G. f. fuscipes in western Kenya. We also determined the influence of colour on the landing response of female and male flies on sticky small targets. Methods Using a series of randomised block experiments, the numbers of tsetse flies caught with sticky small targets were compared with those caught with biconical traps. A negative binomial regression was used to model fly catches. Odds ratios as measures of association between the landing response on the blue or black panel of the sticky small target and the sex of flies were obtained from a multiple logistic regression. Results The results showed that sticky small targets caught 13.5 and 3.6 times more female and male tsetse flies than biconical traps (Z = 9.551, P < 0.0001 and Z = 5.978, P < 0.0001, respectively). Females had a 1.7 times likelihood of landing on the black panel than males (Z = 2.25, P = 0.025). Conclusion This study suggests that sticky small targets are an effective sampling tool for G. f. fuscipes. Therefore, we recommend the use of sticky small targets as an alternative to biconical traps for observational and experimental investigations of G. f. fuscipes.

Tsetse flies (Diptera: Glossinidae) are considered primary cyclical vectors that transmit pathogenic trypanosomes in Africa. They harbour a variety of microbes including Wolbachia, Sodalis and the salivary gland hypertrophy virus (SGHV) which are all vertically transmitted. Knowledge on tsetse microbiome and their interactions may identify novel strategies for tsetse fly and trypanosomiasis control. Area-wide application of such strategies requires an understanding of the natural microbiome frequency in the different species and subspecies of Glossina in their geographical populations. Consequently, this study determined the prevalence of Sodalis, Wolbachia, SGHV and trypanosome infections in Glossina morsitanscentralis from two sites of Western Zambia. We also explored possible associations of the microbes with trypanosome infections. Male G. morsitanscentralis samples were collected from two sites (Lyoni and Lusinina) in Western Zambia. The age structure of the flies at each site was determined using the wing fray method. DNA was extracted from the samples and analyzed for Wolbachia, Sodalis, SGHV and trypanosome presence using PCR. Associations and measures of associations between trypanosome infection and microbes in the fly were determined. The flies from the two locations (Lusinina, n=45 and Lyoni, n=24) had a similar age structure with their median fray category not being significantly different (p=0.698). The overall prevalence of Wolbachia was 72.5% (95% CI: 61.6-83.3%), Sodalis was 15.9% (95% CI: 7.1-24.8%), SGHV was 31.9% (95% CI: 20.6-43.2%) and Trypanosoma species was 23.2% (95% CI: 13-33.4%). The prevalence of Wolbachia was significantly higher in Lusinina than Lyoni (p=0.000). However this was not the case for Sodalis, SGHV and Trypanosoma species. Despite the low number of flies that were positive for both trypanosome and Sodalis (6; 8.7%), a statistically significant association (p=0.013; AOR 6.2; 95% CI: 1.5-25.8) was observed in G. morsitanscentralis. The study showed that the prevalence of microbiota may vary within the same species of the tsetse depending on the geographical location as was the case of Wolbachia. Further it showed that infection with Sodalis could affect vector competence. The study concludes that Sodalis could be an ideal candidate for symbiont-mediated trypanosomiasis control interventions in G. morsitanscentralis.

Trypanocides will continue to play an important role in the control of tsetse fly transmitted trypanosomosis now and in the near future. The drugs are mostly administered by farmers without any veterinary supervision leading to misuse and under dosing of medication, and these could be factors that promote trypanocidal drug resistance (TDR) development. In order to delay or prevent TDR, the Food and Agriculture Organization (FAO) recommended guidelines on trypanocide use. It is not known if these recommended guidelines are adhered to in Itezhi tezhi district of Zambia. A survey was undertaken to examine how socio-economic and environmental factors were associated with adherence to the recommended guidelines on trypanocide use in Itezhi tezhi, Central Zambia. Ninety farmers who use trypanocides were interviewed using a questionnaire to collect their socio-economic characteristics (age, education in years, cattle herd size, competence on trypanocide use and their access to extension on trypanocide use) and trypanocide usage practices while crush pens which they use were stratified according to location, whether in the Game Management Area (GMA) (Mutenda, Itumbi, Kapulwe and Banachoongo) or non-GMA (Iyanda, New Ngoma and Shinampamba) as an environmental factor. Associations and measures of associations to adherence of FAO guidelines were determined. The results showed that 25.6% of the farmers adhered to guidelines by FAO on trypanocide use and that none of the socio-economic factors under investigation were significantly associated with it. Further the farmers that used crush pens that were in the GMA had an 80% reduction in the likelihood of adhering to the FAO guidelines on trypanocide use than those that used crush pens in the non-GMA (AOR 0.20, 95% CI: 0.05–0.81, P = 0.02). There was low adherence to the recommended FAO guidelines on trypanocide use and it was associated with the location of the crush pen whether in the GMA or not, as an environmental factor. With farmers in the GMA less likely to adhere to FAO guidelines than those in the non-GMA, we recommend an integrated approach of measures to control trypanosomosis in the GMA of Itezhi tezhi to lessen overuse of trypanocides by the farmers.

Displacement rates of tsetse affect performance of targets during vector control. Fly size, one of the indicators of population structure usually obtained from wing measurement, is among the determinants of displacement rates. Although recovery of tsetse in previous intervention areas has been widely reported, the population structure of tsetse that recover is rarely evaluated despite being associated with displacements rates. Previously, intervention trials had reduced tsetse densities by over 90% from3 flies/trap/day to1fly/trap/day on Big Chamaunga and Manga islands of Lake Victoria in western Kenya. In this study, we assessed the recovery in densities of Glossina fuscipes fuscipes on the two islands and evaluated the effects vector control might have on the population structure. A before and after intervention study was undertaken on four islands of Lake Victoria in western Kenya; Small and Big Chamaunga, Manga and Rusinga Islands, two of which tsetse control intervention had previously been undertaken. Three years after intervention average G. f. fuscipes catches in biconical traps were estimated on each island. Wing centroid size (CS) (a measurement of fly size) and shape, indicators of the population structure of flies from the four islands were compared using geometric morphometric analyses. CS and shape of available female but not male tsetse wings obtained before the intervention trial on Big and Small Chamaunga islands were compared with those from the same islands after the intervention trial. G. f. fuscipes apparent density on the previous intervention islands were9 flies/trap/day. Irrespective of sex, wing shape did not isolate tsetse based on their islands of origin. The fly size from Big and Small Chamaunga did not differ significantly before intervention trials (P = 0.728). However, three years after the intervention flies from Big Chamaunga were significantly smaller than those from Small Chamaunga (P 0.003). Further, there was an increase in the divergence of wing morphology between flies collected from Big Chamaunga and those from Small Chamaunga after tsetse control. In conclusion, even though populations are not isolated, vector control could influence the population structure of tsetse by exerting size and wing morphology differential selection pressures. Therefore, we recommend further studies to understand the mechanism behind this as it may guide future vector control strategies.