Tungiasis is a neglected tropical disease caused by skin-penetrating female Tunga penetrans fleas. Although tungiasis causes severe health problems, its ecology is poorly understood and morphological descriptions of the larvae are unavailable. To identify T. penetrans immature stages and sites where they develop, diagnostic PCRs are required. However, flea larvae feed on soil organic matter rich in PCR inhibitors. Here, three DNA preparation methods, including a soil DNA kit that removes inhibitors, a simple ammonium acetate precipitation approach (AmAcet) and a crude lysate of larvae (CL), were combined with amplification by the highly processive FIREPol^® Taq or the inhibitor-resistant Phusion^® polymerase. Independent of the polymerase used, the frequency of successful amplification, C_q values and PCR efficacies for the low-cost CL and AmAcet methods were superior to the commercial kit for amplification of a 278 bp partial internal transcribed spacer-2 (ITS-2) and a 730 bp pan-Siphonaptera cytochrome oxidase II PCR. For the CL method combined with Phusion^® polymerase, the costs were approximately 20-fold lower than for the methods based on the soil DNA kit, which is a considerable advantage in resource-poor settings. The ITS-2 PCR did not amplify Ctenocephalides felis genomic or Tunga trimammilata ITS-2 plasmid DNA, meaning it can be used to specifically identify T. penetrans.

Tungiasis is a neglected tropical disease caused by skin-penetrating female Tunga penetrans fleas. Although tungiasis causes severe health problems, its ecology is poorly understood and morphological descriptions of larvae are unavailable. To identify T. penetrans immature stages and sites where they develop, diagnostic PCRs are required. However, flea larvae feed on soil organic matter rich in PCR inhibitors. Here, three DNA preparation methods, a soil DNA kit removing inhibitors, a simple ammonium acetate precipitation approach (AmAcet) and a crude lysate of larvae (CL), were combined with amplification by the highly processive FIREPol® Taq or the inhibitor-resistant Phusion® polymerase. Independent of the polymerase used, frequency of successful amplification, Cq values and PCR efficacies for the low-cost CL and AmAcet methods were superior to the commercial kit for amplification of a 278 bp partial internal transcribed spacer-2 (ITS-2) and a 730 bp pan-Siphonaptera cytochrome oxidase I PCR. For the CL method combined with Phusion® polymerase, costs were approximately 20-fold lower than for methods based on the soil DNA kit, which is a considerable advantage in resource-poor settings. The ITS-2 PCR did not amplify Ctenocephalides felis genomic or Tunga trimammilata ITS-2 plasmid DNA allowing it to be used to specifically identify T. penetrans.

The Centers for Diseases Control (CDC) light trap is widely used for malaria vector surveillance, but its acquisition logistics pose challenges in Africa. Evaluating new traps can improve surveillance tools. This study compared the efficiency of the BG-Pro UV and Silver Bullet 2.1 UV (SB 2.1 UV) against the UV LED CDC trap in western Kenya’s rice irrigation area. The traps were tested indoors in eight houses over 64 nights. Light properties and fan speed were analyzed using spectrometry and an anemometer. The BG-Pro UV trap performed better than the UV LED CDC trap for An. gambiae s.l. (RR 2.0, 95% CI 0.9–3.9) and An. funestus s.l. (RR 3.5, 95% CI 1.9–6.4). The SB 2.1 UV trap was more effective in capturing An. gambiae s.l. (RR 4.3, 95% CI 2.5–7.3) and An. funestus s.l. (RR 7.1, 95% CI 3.9–13.1), and also caught three times more Culex spp. (RR 2.7, 95% CI 1.2–6.0). SB 2.1 UV had the highest downstream force, and all traps emitting UV-A light had consistent wavelengths. Overall, the BG-Pro and SB 2.1 traps’ trapping efficiency was three to six times more than the CDC trap, making them promising surveillance tools, particularly in low-density malaria settings.

The Centres for Diseases Control (CDC) light trap is widely used for malaria vector surveillance, but its acquisition logistics pose challenges in Africa. Evaluating new traps can improve surveillance tools. This study compared the efficiency of BG-Pro UV and Silver Bullet 2.1 UV (SB 2.1 UV) against the UV LED CDC trap in western Kenya’s rice irrigation area. The traps were tested indoors in eight houses over 64 nights. Light properties and fan speed were analysed using spectrometry and an anemometer. The BG-Pro UV trap performed better than the UV LED CDC trap for An. gambiae s.l. (RR 2.0, 95% CI 0.9-3.9) and An. funestus s.l. (RR 3.5, 95% CI 1.9-6.4). The SB 2.1 UV trap was more effective in capturing An. gambiae s.l. (RR 4.3, 95% CI 2.5-7.3) and An. funestus s.l. (RR 7.1, 95% CI 3.9-13.1), and also caught three times more Culex spp. (RR 2.7, 95% CI 1.2-6.0). SB 2.1 UV had the highest downstream force, with and all traps emitting UV-A light with consistent wavelengths. Overall, BG-Pro and SB 2.1 traps trapping efficiency was 3–6 times more than the CDC trap, making them promising surveillance tools, particularly in low-density malaria settings.