BackgroundForecariah is one of the most active foci of gambiense human African trypanosomiasis (gHAT) in Guinea. The study aimed to evaluate the impact of a vector control intervention carried out in this focus from 2018 to 2021. To achieve this, thousands of deltamethrin-impregnated blue tiny targets were deployed annually.Methodology and principal findingsThirty-one sentinel traps were used to assess the following entomological and parasitological parameters: (i) the apparent density (AD) of tsetse flies, monitored twice per year; (ii) the tsetse fly infection rates, evaluated once per year; and (iii) the trypanosome species, determined by molecular approaches before and after three years of vector control. Prior to the VC campaign, significantly higher densities of Glossina palpalis gambiensis were observed in the mangroves (AD = 17.2 F/T/D) than in mainland areas (AD = 6.2 F/T/D). Just 18 months after the start of the VC campaign, the maximum reduction in AD was reached (mean AD = 1.1 F/T/D; p < 0.001; 92% reduction) and was maintained throughout the campaign. Prior to VC, 18 out of 31 (58%) sentinel traps presented at least one infected fly observed by microscopy, whereas only two out of 31 (6%; p = 0.002) traps were positive after three years of VC. Molecular analysis of the tsetse midgut revealed positivity for T. brucei sl DNA in 45.2% of the traps prior to the VC baseline. T. congolense and T. vivax were observed in 6.5% of traps. Although one T. b. gambiense infection, which is responsible for human disease, was detected three years after the introduction of vector control, a significant decrease in T. brucei sl PCR positivity was observed (45.2% versus 22.6%; p = 0.05), whereas a slight increase was observed for T. vivax (6.5% versus 16.1%), while T. congolense trap positivity (6.5%) remained stable.ConclusionIn Forecariah, the prevalence of the disease was reduced by 76% during the study period. Hence, these results further emphasize that implementing vector control alongside medical control is an effective way of reducing the transmission of parasites to humans.

The Republic of Guinea has faced an important challenge with human African trypanosomiasis (HAT), which was endemic over the last century. After initial control in the 1960s-1970s, HAT resurged in the 1990s along the Guinean coast, driven by economic and demographic pressures on the mangrove ecosystem. In response, the Guinean government established a national control program in 2002, focusing on medical mass screenings. In 2012, vector control using tiny targets was introduced in the East Boffa focus to reduce fly density and human-vector contact. However, the Ebola epidemic from 2013 to 2016 disrupted these efforts, leading to a reliance on passive screening. Resuming screenings in 2016-2017 revealed increased cases in all foci except the East Boffa area, where vector control had been effective. Vector control continued during the SARS-CoV2 pandemic and at the same time targeted door-to-door screenings were introduced to target high-risk individuals. Since 2018, around 30,000 at-risk individuals have been screened annually. These strategies reduced the total number of new cases below 1 per 10,000 inhabitants in endemic areas over the period 2019-2023, allowing to validate the elimination of HAT as a public health problem. The Guinean team and partners then focused on systematic spatial monitoring of patients and community engagement in vector control. The program also integrates control of other neglected tropical diseases and addresses new research questions, especially about anatomical and animal reservoirs for parasites. These efforts, combined with implementation of improved diagnostic tests and new oral treatments, through active involvement in multiple clinical trials and studies, now aim to interrupt HAT transmission by 2030.

Human leukocyte antigen-G (HLA-G) is an immunomodulatory molecule known to play a crucial role in immune tolerance and regulation. In the context of human African trypanosomiasis (HAT), higher soluble HLA-G levels were detected in the plasma of confirmed cases, representing a serological marker of T. b. gambiense infection. As trypanosomes also invade extravascular tissues, especially the skin, this study explored the potential role of HLA-G in the dermal immune response during T. b. gambiense infection. Blood and skin samples from 50 seronegative individuals, 45 seropositive suspects and 36 confirmed HAT cases, collected between 2018 and 2022 in endemic foci of Guinea and Côte d'Ivoire, were analyzed. Plasmatic and dermal levels of HLA-G proteins were quantified by ELISA and immuno-histochemistry, respectively, and compared to the trypanosome detection results in the same samples. The implication of soluble HLA-G plasma level as a biomarker of T. b. gambiense infection was confirmed. In the dermis, HLA-G isoforms were expressed either with a granular distribution or with in diffuse halos. Granular patterns of dermal HLA-G were directly associated with the presence of trypanosomes in the dermis. The presence of diffuse halos was correlated to higher sHLA-G levels in the plasma. In total, this study provides the first evidence of the involvement of HLA-G in the extravascular immune response against parasites, especially in the skin. It shows that HLA-G distribution in the extravascular compartment also represents a biomarker of trypanosome infection.

In the population at risk of gambiense human African trypanosomiasis (gHAT), the prevalence of extravascular parasite carriage remains unclear. Here, we conducted an observational clinical study in the hypo-endemic gHAT foci of Sinfra and Bonon in Côte d'Ivoire from 2019 to 2022. A total of 74 individuals were enrolled, including 45 suspects previously found positive at least once in a serological test for gHAT and followed by the national elimination programme of Côte d'Ivoire, as well as 29 seronegative controls. No significant differences between groups were observed for any epidemiological parameters and any clinical parameters at enrolment. Whereas trypanosome DNA was detected in the blood of 0/29 controls and 2/45 suspects, the presence of extravascular dermal trypanosomes was confirmed by immuno-histochemistry (fixed trypanosome cells) and/or PCR (trypanosome DNA) in about 1/3 of the suspects (14/45, 31%). However, no gambiense-specific test was found positive in the present study. Hence, the skin could represent an anatomical reservoir for African trypanosomes sustaining a low level of transmission in hypo-endemic foci.

In the mangroves of Guinea, where the most active foci of human African trypanosomiasis in West Africa are located, vector control against tsetse flies using insecticide-impregnated Tiny Targets was first introduced in 2012. While annual deployments of Tiny Targets have resulted in an overall 90

Objectif : Évaluer la faisabilité du lancement du nouveau médicament contre la THA et son utilisation dans les trois foyers endémiques de Guinée. Méthodologie : Une étude transversale analytique a été réalisée pour évaluer la faisabilité de cette mise en route dans les foyers de Boffa, Dubréka et Forécariah situés sur le littoral Guinéen. Résultats : Les résultats de cette étude ont montré que (i) près d’un répondant sur trois 27,5 % avait une bonne connaissance sur le mode de transmission de la THA, (ii) la moitié des répondants 49,6 % avait entendu parler d’un nouveau traitement de la THA. La grande majorité de répondants reconnaissait qu’il a les avantages de ne pas nécessiter de ponction lombaire 61,4 % et d’être gratuit 38,6 %. Presque tous les répondants accepteraient volontiers un traitement oral de la THA 99,3 % et seraient prêts à le prendre même à domicile 98,3 %. Beaucoup parmi eux se sentiraient moins stigmatisés en suivant ce traitement. Conclusion : Bien que les connaissances requises, l’attitude favorable, et les pratiques disposant à l’adoption de ce nouveau traitement aient été différentes en fonction du site d’étude et du niveau de scolarisation, les répondants ont largement indiqué qu’ils adopteraient ce nouveau traitement avec confiance.

Objective: To evaluate the feasibility of launching the new human African trypanosomiasis (HAT) drug and its use in the three endemic foci in Guinea. Methodology: An analytical cross-sectional study was conducted to assess the feasibility of this implementation in the foci of Boffa, Dubr & eacute;ka, and For & eacute;cariah located on the Guinean coast. Results: The results of this study showed that (i) nearly one in three respondents (27.5%) had good knowledge of how HAT is transmitted, and (ii) half of the respondents (49.6%) had heard of a new treatment for HAT. Most of the respondents recognized that it has the advantages of not requiring a lumbar puncture (61.4%) and of being free of charge (38.6%). Almost all respondents would readily accept oral HAT treatment (99.3%) and would even be willing to take it at home (98.3%). Many of them would feel less stigmatized by following this treatment. Conclusion: Although the knowledge required, favorable attitude, and practices amenable to adopting this new treatment varied, depending on the study site and level of education, the respondents largely indicated that they would be confident in adopting this new treatment.

Animal African trypanosomosis (AAT), caused by protist parasites of the genus Trypanosoma, puts upward of a million head of livestock at risk across 37 countries in Africa. The economic impact of AAT and the presence of human-infectious trypanosomes in animals place a clear importance on improving diagnostics for animal trypanosomes to map the distribution of the veterinary parasites and identify reservoirs of human-infectious trypanosomes. We have adapted the CRISPR-based detection toolkit SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) for trypanosomatid parasites responsible for AAT (SHERLOCK4AAT) including Pan-trypanosomatid, Trypanozoon, T. vivax, T. congolense, T. theileri, T. simiae and T. suis assays. To test the applicability of this technique in the field, we analysed dried blood spots collected from 200 farm and 224 free-ranging pigs in endemic and historical human African trypanosomiasis foci in Guinea and Cote d Ivoire, respectively. The results revealed that SHERLOCK4AAT can detect and discriminate between trypanosome species involved in multiple infections with a high sensitivity. 62.7 % [58.1, 67.3] of pigs were found infected with at least one trypanosome species. T. brucei gambiense, a human-infectious trypanosome, was found in one animal at both sites, highlighting the risk that these animals may act as persistent reservoirs. These data suggest that, due to their proximity to humans and their attractiveness to tsetse flies, pigs could act as sentinels to monitor T. b. gambiense circulation using the SHERLOCK4AAT toolbox. ### Competing Interest Statement The authors have declared no competing interest.

Background . Although Guinea has eliminated human African trypanosomiasis (HAT) as a public health problem, challenges remain in identifying the remaining cases, particularly among young men who work in mangrove areas and are highly exposed to the tsetse fly vector. This study aimed to evaluate the effectiveness of targeted sensitisation campaigns for young men in improving their participation in active screening campaigns for HAT in Guinea. Method . A door-to-door screening campaign in Boffa, Guinea, coincided with the 2022 Football World Cup. Football matches were projected to take advantage of the opportunity to raise awareness of HAT among the population. Twenty-seven villages were selected for the screening, and four villages where HAT cases had recently been diagnosed were chosen to broadcast World Cup matches, followed by a 30-minute film about HAT in the local language. Demographic data of those who attended HAT screening was collected using an existing Open Data Kit (ODK) system. Results . The results indicated significant interest in the awareness-raising events, with over 500 people attending over the four days of broadcasting. Comparing participation rates between World Cup villages (WCVs) and non-WCVs, a higher proportion of males aged 8–27 participated in the WCVs (17.7%) than in non-WCVs (14.6%). This difference was not observed for females in the same age group. Furthermore, compared to data from the previous year, there was an increase in the relative participation of young males, rising from 16.4% to 19.6%. Conclusion . Innovative strategies, such as using football matches to raise awareness, are essential for identifying remaining HAT cases and reaching at-risk populations. Football holds significant cultural and social importance in Guinean villages, making it an effective platform for raising awareness of HAT. Expanding such strategies could further increase awareness and participation in screening campaigns, accelerating the elimination of HAT transmission.

Animal African trypanosomosis (AAT), caused by protist parasites of the genus Trypanosoma, puts upward of a million head of livestock at risk across 37 countries in Africa. The economic impact of AAT and the presence of human-infectious trypanosomes in animals place a clear importance on improving diagnostics for animal trypanosomes to map the distribution of the veterinary parasites and identify reservoirs of human-infectious trypanosomes. We have adapted the CRISPR-based detection toolkit SHERLOCK (Specific High-sensitivity Enzymatic Reporter unLOCKing) for trypanosomatid parasites responsible for AAT (SHERLOCK4AAT) including Pan-trypanosomatid, Trypanozoon, T. vivax, T. congolense, T. theileri, T. simiae, and T. suis assays. To test the applicability of this technique in the field, we analysed dried blood spots collected from 200 farm and 224 free-ranging pigs in endemic and historical human African trypanosomiasis foci in Guinea and Côte d’Ivoire, respectively. The results revealed that SHERLOCK4AAT can detect and discriminate between trypanosome species involved in multiple infections with a high sensitivity. 62.7% [58.1, 67.3] of pigs were found infected with at least one trypanosome species. T. brucei gambiense, a human-infectious trypanosome, was found in one animal at both sites, highlighting the risk that these animals may act as persistent reservoirs. These data suggest that, due to their proximity to humans and their attractiveness to tsetse flies, pigs could act as sentinels to monitor T. b. gambiense circulation using the SHERLOCK4AAT toolbox.

Background. The study aimed to evaluate the effectiveness of sensitization events targeting young males to improve their participation in active screening campaigns for Human African Trypanosomiasis (HAT), also known as sleeping sickness, in Guinea. Despite progress towards eliminating HAT as a public health problem, challenges remain in detecting the last remaining cases, particularly among individuals, especially young men, who work in mangroves and are highly exposed to the tsetse fly vector. Method. During the 2022 Football World Cup, coinciding with a door-to-door screening campaign in the Boffa focus of Guinea, football matches were used as an opportunity to sensitize the population. Twenty-seven villages were selected for screening, and four villages with recently diagnosed HAT cases were chosen to broadcast World Cup matches followed by a 30-minute sensitization movie about HAT in the local language. Demographic data were collected using an Open Data Kit system. Results. The results indicated significant interest in the sensitization events, with over 500 attendees at all during the four days of broadcasting. A comparison of participation rates between World Cup villages (WCV) and non-WCV villages showed a higher proportion of males aged 8 to 27 participating in the WCV (17.7%) compared to non-WCV villages (14.6%). This difference was not observed for females of the same age group. Furthermore, compared to data from the previous year, there was an increase in the relative participation of young males from 16.4% to 19.6%. Conclusion. Innovative strategies, such as using football matches for sensitization, are essential for reaching at-risk populations and identifying remaining HAT cases. Football holds significant cultural and social importance in Guinean villages, making it an effective platform for HAT sensitization efforts. Scaling up such strategies could further enhance awareness and participation in screening campaigns, ultimately aiding in the elimination of HAT transmission.

Human African trypanosomosis (HAT), also known as sleeping sickness, is still a major concern in endemic countries. Its cyclical vector are biting insects of the genus Glossina or tsetse flies. In Guinea, the mangrove ecosystem contains the main HAT foci of Western Africa. There, the cyclical vector is Glossina palpalis gambiensis . A still ongoing vector control campaign (VCC) started in 2011 in the focus of Boffa, using tiny targets, with a 79% tsetse density reduction in 2016 and significant impact on the prevalence of the disease (from 0.3% in 2011 to 0.11% in 2013, 0.0352% in 2016 and 0.0097% in 2019). To assess the sustainability of these results, we have studied the impact of this VCC on the population biology of G. p. gambiensis in Boffa. We used the genotyping at 11 microsatellite markers and population genetic tools of tsetse flies from different sites and at different dates before and after the beginning of the VCC. In variance with a significant impact of VCC on the apparent densities of flies captured in the traps deployed, the global population of G. p. gambiensis displayed no variation of the sex-ratio, no genetic signature of control, and behaved as a very large population occupying the entire zone. This implies that targets deployment efficiently protected the human populations locally, but did not impact tsetse flies where targets cannot be deployed and where the main tsetse population exploits available resources. We thus recommend the pursuit of vector control measures with the same strategy, through the joint effect of VCC and medical surveys and treatments, in order to protect human populations from HAT infections until the disease can be considered as entirely eradicated from the focus.

Background:Serological screening tests play a crucial role to diagnose gambiense human African trypanosomiasis (gHAT). Presently, they preselect individuals for microscopic confirmation, but in future “screen and treat” strategies they will identify individuals for treatment. Variability in reported specificities, the development of new rapid diagnostic tests (RDT) and the hypothesis that malaria infection may decrease RDT specificity led us to evaluate the specificity of 5 gHAT screening tests. Method: During active screening, venous blood samples from 1095 individuals from Côte d’Ivoire and Guinea were tested consecutively with commercial (Bioline HAT 2.0, HAT Sero-K-SeT, CATT) and prototype (HAT Sero-K-SeT 2.0, DCN) gHAT screening tests and with a malaria RDT. Individuals with ≥ 1 positive gHAT screening test underwent microscopy and further immunological (trypanolysis, indirect ELISA) and molecular laboratory tests (conventional PCR, SHERLOCK, Trypanozoon S²-RT-qPCR, SNP RT-qPCR). Microscopic trypanosome detection confirmed gHAT, while other individuals were considered gHAT free. Results: One gHAT case was diagnosed. Overall test specificities (n=1094) were: CATT 98.9% (98.1-99.4%); HAT Sero-K-SeT 86.7% (84.5-88.5%); Bioline HAT 2.0 82.1% (79.7-84.2%); DCN HAT RDT 78.2% (75.7-80.6%); and HAT Sero-K-SeT 2.0 78.4% (75.9-80.8%). In malaria positives, gHAT screening tests appeared less specific, but the difference was significant only in Guinea for Bioline HAT 2.0 and HAT Sero-K-Set 2.0. The specificities of immunological and molecular laboratory tests in gHAT seropositives were 98.7-100% (n=399) and 93.0-100% (n=302), respectively. Among 44 laboratory test positives, only the confirmed gHAT patient and one screening test seropositive combined immunological and molecular laboratory test positivity. Conclusions:Although a minor effect of malaria cannot be excluded, gHAT RDT specificities are far below the 95% minimal specificity stipulated by the WHO target product profile for a simple diagnostic tool to identify individuals eligible for treatment. Unless specificity is improved, an RDT-based “screen and treat” strategy would result in massive overtreatment. In view of their inconsistent results, additional comparative evaluations of the diagnostic performance of laboratory tests are indicated for better identifying, among screening test positives, those at increased suspicion for gHAT. Trial registration: The trial was retrospectively registered under NCT05466630 in clinicaltrials.gov on July 15 2022.

Trypanosoma brucei gambiense (Tbg) group 2 is a subgroup of trypanosomes able to infect humans and is found in West and Central Africa. Unlike other agents causing sleeping sickness, such as Tbg group 1 and Trypanosoma brucei rhodesiense, Tbg2 lacks the typical molecular markers associated with resistance to human serum. Only 36 strains of Tbg2 have been documented, and therefore, very limited research has been conducted despite their zoonotic nature. Some of these strains are only available in their procyclic form, which hinders human serum resistance assays and mechanistic studies. Furthermore, the understanding of Tbg2’s potential to infect tsetse flies and mammalian hosts is limited. In this study, 165 Glossina palpalis gambiensis flies were experimentally infected with procyclic Tbg2 parasites. It was found that 35 days post-infection, 43 flies out of the 80 still alive were found to be Tbg2 PCR-positive in the saliva. These flies were able to infect 3 out of the 4 mice used for blood-feeding. Dissection revealed that only six flies in fact carried mature infections in their midguts and salivary glands. Importantly, a single fly with a mature infection was sufficient to infect a mammalian host. This Tbg2 transmission success confirms that Tbg2 strains can establish in tsetse flies and infect mammalian hosts. This study describes an effective in vivo protocol for transforming Tbg2 from procyclic to bloodstream form, reproducing the complete Tbg2 cycle from G. p. gambiensis to mice. These findings provide valuable insights into Tbg2’s host infectivity, and will facilitate further research on mechanisms of human serum resistance.

The skin is an anatomical reservoir for African trypanosomes, yet the prevalence of extravascular parasite carriage in the population at risk of gambiense Human African Trypanosomiasis (gHAT) remains unclear. Here, we conducted a prospective observational cohort study in the HAT foci of Forecariah and Boffa, Republic of Guinea. Of the 18,916 subjects serologically screened for gHAT, 96 were enrolled into our study. At enrolment and follow-up visits, participants underwent a dermatological examination and had blood samples and superficial skin snip biopsies taken for examination by molecular and immuno-histological methods. In seropositive individuals, dermatological symptoms were significantly more frequent as compared to seronegative controls. Trypanosoma brucei DNA was detected in the blood of 67% of confirmed cases (22/33) and 9% of unconfirmed seropositive individuals (3/32). However, parasites were detected in the extravascular dermis of up to 71% of confirmed cases (25/35) and 41% of unconfirmed seropositive individuals (13/32) by PCR and/or immuno-histochemistry. Six to twelve months after treatment, trypanosome detection in the skin dropped to 17% of confirmed cases (5/30), whereas up to 25% of unconfirmed, hence untreated, seropositive individuals (4/16) were still found positive. Dermal trypanosomes were observed in subjects from both transmission foci, however, the occurrence of pruritus and the PCR positivity rates were significantly higher in unconfirmed seropositive individuals in Forecariah. The lower sensitivity of superficial skin snip biopsies appeared critical for detecting trypanosomes in the basal dermis. These results are discussed in the context of the planned elimination of gHAT.

Abstract Background Serological screening tests play a crucial role to diagnose gambiense human African trypanosomiasis (gHAT). Presently, they preselect individuals for microscopic confirmation, but in future “screen and treat” strategies they will identify individuals for treatment. Variability in reported specificities, the development of new rapid diagnostic tests (RDT) and the hypothesis that malaria infection may decrease RDT specificity led us to evaluate the specificity of 5 gHAT screening tests. Methods During active screening, venous blood samples from 1095 individuals from Côte d’Ivoire and Guinea were tested consecutively with commercial (CATT, HAT Sero-K-SeT, Abbott Bioline HAT 2.0) and prototype (DCN HAT RDT, HAT Sero-K-SeT 2.0) gHAT screening tests and with a malaria RDT. Individuals with ≥ 1 positive gHAT screening test underwent microscopy and further immunological (trypanolysis with T.b. gambiense LiTat 1.3, 1.5 and 1.6; indirect ELISA/T.b. gambiense; T.b. gambiense inhibition ELISA with T.b. gambiense LiTat 1.3 and 1.5 VSG) and molecular reference laboratory tests (PCR TBRN3, 18S and TgsGP; SHERLOCK 18S Tids, 7SL Zoon, and TgsGP; Trypanozoon S2-RT-qPCR 18S2, 177T, GPI-PLC and TgsGP in multiplex; RT-qPCR DT8, DT9 and TgsGP in multiplex). Microscopic trypanosome detection confirmed gHAT, while other individuals were considered gHAT free. Differences in fractions between groups were assessed by Chi square and differences in specificity between 2 tests on the same individuals by McNemar. Results One gHAT case was diagnosed. Overall test specificities (n = 1094) were: CATT 98.9% (95% CI: 98.1–99.4%); HAT Sero-K-SeT 86.7% (95% CI: 84.5–88.5%); Bioline HAT 2.0 82.1% (95% CI: 79.7–84.2%); DCN HAT RDT 78.2% (95% CI: 75.7–80.6%); and HAT Sero-K-SeT 2.0 78.4% (95% CI: 75.9–80.8%). In malaria positives, gHAT screening tests appeared less specific, but the difference was significant only in Guinea for Abbott Bioline HAT 2.0 (P = 0.03) and HAT Sero-K-Set 2.0 (P = 0.0006). The specificities of immunological and molecular laboratory tests in gHAT seropositives were 98.7–100% (n = 399) and 93.0–100% (n = 302), respectively. Among 44 reference laboratory test positives, only the confirmed gHAT patient and one screening test seropositive combined immunological and molecular reference laboratory test positivity. Conclusions Although a minor effect of malaria cannot be excluded, gHAT RDT specificities are far below the 95% minimal specificity stipulated by the WHO target product profile for a simple diagnostic tool to identify individuals eligible for treatment. Unless specificity is improved, an RDT-based “screen and treat” strategy would result in massive overtreatment. In view of their inconsistent results, additional comparative evaluations of the diagnostic performance of reference laboratory tests are indicated for better identifying, among screening test positives, those at increased suspicion for gHAT. Trial registration The trial was retrospectively registered under NCT05466630 in clinicaltrials.gov on July 15 2022. Graphical Abstract

Strategies to detect Human African Trypanosomiasis (HAT) cases rely on serological screening of populations exposed to trypanosomes. In Guinea, mass medical screening surveys performed with the Card Agglutination Test for Trypanosomiasis have been progressively replaced by door-to-door approaches using Rapid Diagnostic Tests (RDTs) since 2016. However, RDTs availability represents a major concern and medical teams must often adapt, even in the absence of prior RDT performance evaluation. For the last 5 years, the Guinean HAT National Control Program had to combine three different RDTs according to their availability and price: the SD Bioline HAT (not available anymore), the HAT Sero-K-SeT (most expensive), and recently the Abbott Bioline HAT 2.0 (limited field evaluation). Here, we assess the performance of these RDTs, alone or in different combinations, through the analysis of both prospective and retrospective data. A parallel assessment showed a higher positivity rate of Abbott Bioline HAT 2.0 (6.0%, n = 2,250) as compared to HAT Sero-K-SeT (1.9%), with a combined positive predictive value (PPV) of 20.0%. However, an evaluation of Abbott Bioline HAT 2.0 alone revealed a low PPV of 3.9% (n = 6,930) which was surpassed when using Abbott Bioline HAT 2.0 in first line and HAT Sero-K-SeT as a secondary test before confirmation, with a combined PPV reaching 44.4%. A retrospective evaluation of all 3 RDTs was then conducted on 189 plasma samples from the HAT-NCP biobank, confirming the higher sensitivity (94.0% [85.6-97.7%]) and lower specificity (83.6% [76.0-89.1%]) of Abbott Bioline HAT 2.0 as compared to SD Bioline HAT (Se 64.2% [52.2-74.6%]-Sp 98.4% [94.2-99.5%]) and HAT Sero-K-SeT (Se 88.1% [78.2-93.8%]-Sp 98.4% [94.2-99.5%]). A comparison of Abbott Bioline HAT 2.0 and malaria-RDT positivity rates on 479 subjects living in HAT-free malaria-endemic areas further revealed that a significantly higher proportion of subjects positive in Abbott Bioline HAT 2.0 were also positive in malaria-RDT, suggesting a possible cross-reaction of Abbott Bioline HAT 2.0 with malaria-related biological factors in about 10% of malaria cases. This would explain, at least in part, the limited specificity of Abbott Bioline HAT 2.0. Overall, Abbott Bioline HAT 2.0 seems suitable as first line RDT in combination with a second HAT RDT to prevent confirmatory lab overload and loss of suspects during referral for confirmation. A state-of-the-art prospective comparative study is further required for comparing all current and future HAT RDTs to propose an optimal combination of RDTs for door-to-door active screening.

Although studies on African Trypanosomiases revealed a variety of trypanosome species in the blood of various animal taxa, animal reservoirs of Trypanosoma brucei gambiense and anatomical niches such as skin have been overlooked in most epidemiological settings. This study aims to update epidemiological data on trypanosome infections in animals from human African trypanosomiasis (HAT) foci of Cameroon. Blood and skin snips were collected from 291 domestic and wild animals. DNA was extracted from blood and skin snips and molecular approaches were used to identify different trypanosomes species. Immunohistochemical analyses were used to confirm trypanosome infections in skin snips. PCR revealed 137 animals (47.1%) with at least one trypanosome species in the blood and/or in the skin. Of these 137 animals, 90 (65.7%) and 32 (23.4%) had trypanosome infections respectively in the blood and skin. Fifteen (10.9%) animals had trypanosome infections in both blood and skin snip. Animals from the Campo HAT focus (55.0%) were significantly (X2 = 17.6; P< 0.0001) more infected than those (29.7%) from Bipindi. Trypanosomes of the subgenus Trypanozoon were present in 27.8% of animals while T. vivax, T. congolense forest type and savannah type were detected in 16.5%, 10.3% and 1.4% of animals respectively. Trypanosoma b. gambiense infections were detected in the blood of 7.6% (22/291) of animals. No T. b. gambiense infection was detected in skin. This study highlights the presence of several trypanosome species in the blood and skin of various wild and domestic animals. Skin appeared as an anatomical reservoir for trypanosomes in animals. Despite methodological limitations, pigs, sheep, goats and wild animals were confirmed as potential reservoirs of T. b. gambiense. These animal reservoirs must be considered for the designing of control strategies that will lead to sustainable elimination of HAT.

Human African Trypanosomiasis (HAT) is caused by Trypanosoma brucei which is transmitted by the tsetse fly insect vector (Glossina spp). It is one of the 20 Neglected Tropical Diseases (NTD) listed by the WHO. These diseases affect the poorest and most vulnerable communities, for which the WHO has established a dedicated 2021-2030 roadmap. At the time of Alphonse Laveran, HAT devastated the African continent. In the 1960s, the disease was nearly under control, but it strongly re-emerged in the 1990s. A coordinated effort of all stakeholders, with national control programs as the main actors, a strong contribution of research and important donations by the private sector, allowed to decrease the HAT burden significantly. Since 2018, less than 1000 cases are detected annually. We here review new diagnostics, treatments and vector control tools that have been implemented jointly and successfully in several endemic countries.The next key challenge will be to sustain the gains. Newly emerging research questions include long-term carriage of trypanosomes and adaptation of tools to low prevalence contexts. Challenges out of the research area comprise the continued need of funding, maintenance of dedicated human resources, and the key question of access. Sustainable elimination as "interruption of transmission", which is the 2030 NTD roadmap target, can be reached, if these challenges are solved. We stress the importance of continuing to combine the efforts in the fight against the disease, because sustainable elimination of HAT is the best long-term prevention strategy against re-emergence. As such, HAT elimination can serve as an example for other infectious diseases.