The global prevalence obtained w

The global prevalence obtained with the mAECT performed on 57 CATT-B positive pigs was 93%, ranging from 50% in CIFCI to 100% in Gatifla and TG. Global positivity rates with the other tests ranged from 4% (TVW PCR) to 49% (CATT-B). TBR = TBR1-2 primers, TCF = TCF1-2 primers, + = positive test * TG = Trafla Gottron. + = positive test * TG = Trafla Gottron ** = LiTat 1.5 and 1.6 occurrence was significantly higher (p values<0.001) in PK14 and Gozy comparing to the other study sites. The Glycosylphosphatidylinositol Anchor: A Linchpin for Cell Surface Versatility of Trypanosomatids. A total of 167 pigs were investigated. Conceptualization, Supervision, Methodology, They continue to impact public health and socio-economic development mainly in remote rural areas in Sub-Saharan Africa.

In our study, we observed significant differences in prevalence and in result profiles for several diagnostic tests according to the study sites that are only a few kilometers apart. Serological examination revealed 15 CATT-B positive individuals and 6 CATT-P (2 in PK5, 3 in Gatifla and 1 in Dema) for whom parasitological investigations, TBR PCR and TL were negative. Yes Unit de Recherche Trypanosomoses , Institut Pierre Richet, Bouak, Cte dIvoire, The current situation on animal trypanosomiasis in Africa.

The nested TgsGP PCR applied on these samples were negative suggesting a non-specific amplification during the first PCR round. Pig keeping is becoming increasingly common across sub-Saharan Africa. https://doi.org/10.1371/journal.pntd.0010036.g002. A previous study conducted in the Bonon and Sinfra HAT endemic foci concluded that pigs were the most trypanosome-infected domestic animals, but the presence of T. b. gambiense could not be proven [11]. 2022 Mar 7;9:828111. doi: 10.3389/fvets.2022.828111. https://doi.org/10.1371/journal.pntd.0010036.g005. Careers. These proximity and trophic interactions result on predominant free-ranging pig/tsetse transmission cycles in the vicinity of villages. and T. congolense have been previously described in pigs in the beginning of the 1980s [20], the current situation of AAT is completely unknown in that area. Formal analysis, Unit de Recherche sur les Maladies Vecteurs et Biodiversit, Centre International de Recherche-Dveloppement sur lElevage en zone Subhumide, Bobo-Dioulasso, Burkina Faso, Roles Molecular epidemiological studies on animal trypanosomiases in Ghana. Excluding or evidencing this role using field and experimental studies is crucial to adapt the control strategies and to define indicators to reach the interruption of transmission targeted by WHO in 2030 [2]. Surprisingly, two of these strains were TBR PCR negative, while the six corresponding blood samples were TBR PCR positive (Fig 6).

While T. b. rhodesiense HAT is known to be zoonotic, the role of a domestic or wild animal reservoir in the T. b. gambiense HAT epidemiology is still under debate [3,4].

Methodology, As can be seen in Fig 2, TG and Gozy (and to a lesser extent Dema, Gatifa and CIFCI), sites with exclusive free ranging pigs, represented the most heavily infected sites. The acute form, due to T. b. rhodesiense, occurs in Eastern and Southern Africa, while the chronic form, due to T. b. gambiense, is rampant in Western and Central Africa, causing 98% of HAT cases [2]. Another difficulty relies on the fact that the most exposed animal(s) are often multi-infected as already observed in several studies [11,72]. No significant differences were observed between the study sites for LiTat 1.3. Funding acquisition, Non-specific TgsGP PCR products could explain the reported presence of T. b. gambiense in domestic and wild animals in foci where no or very few HAT cases were reported [41,65,66]. Studies focusing on the deeper genetic knowledge of the circulating strains (by NGS methods) and phenotypical evidences related to resistance toward human serum are currently conducted to solve these questions. mAECT was much more sensitive than BCT. Prev Vet Med. Is the Subject Area "Swine" applicable to this article? Unit de Recherche Trypanosomoses , Institut Pierre Richet, Bouak, Cte dIvoire, Roles Fig 5 shows the global proportions of free-ranging pigs positive to 3 (11%), 2 (29%) and 1 VAT (20%) and the proportions according to the study sites, once more confirming an important heterogeneity between those. Writing review & editing, Roles This is probably due to the fact that free-ranging pigs, roaming freely in the humid and shady areas and in the protected sacred forests near by villages, are highly exposed to Glossina palpalis that shares the same biotope as already observed [11,46]. Given the 2018 historic threshold of 1,000 cases reached [2], the potential role of animals as reservoirs did not prevent to control HAT so far. Supervision, No, Is the Subject Area "Trypanosoma brucei" applicable to this article? Human African trypanosomiasis (HAT), or sleeping sickness, exists in two forms depending on the subspecies of Trypanosoma brucei [1]. These blood samples were tested with the standard buffy coat technique (BCT) [24].

Formal analysis, TgsGP PCR data were added manually. https://doi.org/10.1371/journal.pntd.0010036.g001. All the combinations of TL positive results with the three VAT were observed, but the combination Litat 1.3 negative, LiTat 1.5 positive and LiTat 1.6 positive was the most represented profile with 18 pigs (19%). World Dev. A significant difference was observed between the study sites for all tests. No HAT cases were detected. Five days post inoculation, parasitemia was determined daily by direct microscopic examination (X400) of mouse tail blood [25]. Taking into account both BCT and mAECT results, the global prevalence in free-ranging pigs was 72% (70/97). Moreover, five other strains that gave a low intensity band with the single round TgsGP PCR (S1 Fig) were negative for the nested TgsGP PCR. Funding acquisition, Control of Rhodesian sleeping sickness. Ignoring or under-estimating the importance of this topic may lead to repeat the same mistakes as for the Guinea worm for which the reporting of a canine reservoir has challenged and delayed the eradication process [73]. CIFCI and Gatifla appeared to be less affected by trypanosome infections than the other sites (Table 3). brucei trypanosoma Methodology, In the same region, the Vavoua HAT focus was epidemic at the end of the 1970s [1214] and thanks to both medical and vector control efforts [15,16], less than five HAT cases were reported yearly at the end of the 1990s [17]. While T. brucei s.l. If free-ranging pigs seems to be mostly involved in trypanosome circulation in Cte dIvoire, other animal species appeared to be involved in other countries with differences observed between different HAT foci [44,57] and even within the same focus [42]. All participants were informed of the objective of the study in their own language and signed an informed consent form.

Except for TVW PCR (p-values>0.5), study sites, and probably the husbandry method, together had a highly significant impact on the diagnostic tests results (all p-values<0.003) (S3 Table). The PCR products were visualized by electrophoresis in a 2% agarose gel (MP Biomedical, Eschwege, Germany) stained with Gel Red (Interchim, Montluon, France) and illuminated with UV light. Roles

No, Is the Subject Area "Trypanosoma" applicable to this article? Before With the increase of molecular approaches, presence of T. vivax in pigs has been recently reported [48,49] contrasting with the historical data previously reviewed [50]. LiTat 1.3 and LiTat 1.5 VATs are supposed to be specific for T. b. gambiense, while LiTat 1.6 VAT is expressed in T. b. gambiense and T. b. brucei [34].

The association between these two species, already mentioned in other studies [11,20,39,51,72] may hold epidemiological consequences that would deserve further investigations. The mAECT, known as the most sensitive parasitological test for T. b. gambiense detection in human [58,59], was applied on CATT positive pigs for which infection rates from 50% to 100% were observed. Sample 3 (pig 66), 7 (pig 69), 8 (pig 84), 12 (pig 100), 14 (pig 102) and 16 (pig 104) C1+ and C2+ = positive PCR controls (T. b. gambiense reference stock). https://doi.org/10.1371/journal.pntd.0010036.s004. However, studies conducted in the two HAT foci that are still endemic in the central-western part of the country (Bonon and Sinfra) recently demonstrated that pigs were particularly infected by trypanosomes, although infections with T. b. gambiense could not be proven due to a lack of reliability of the tools used [11]. Methodology, https://doi.org/10.1371/journal.pntd.0010036.t003. We showed that T. brucei s.l. The low infection rates observed in pigs in pen could be because pigsties are located inside or close to the villages far away from the tsetse flies favorable biotopes, and are thus probably less exposed. Yes Acta Trop. Writing original draft, Roles Supervision, Accessibility Writing review & editing, Affiliation The human active screening was conducted within the framework of medical surveys and epidemiological surveillance activities supervised by the HAT National Control Program (HAT NCP). DNA from 500 L of BC was extracted using the DNeasy Blood and Tissue kit (Qiagen, Valencia, CA, USA) following manufacturers instructions. This was a deliberate choice regarding the single round and nested PCR results firstly obtained on purified DNA from isolated strains. Second, a TgsGP-related gene (Tb10.v4.0178) that shared 81% homology with TgsGP has been described in T. b. brucei [62,63]. Yes Methodology, Roles Alternatively we cannot exclude that TgsGP nested PCR may detect very low level of DNA in animals, that does not mean any active transmission to humans.

Serological and parasitological results obtained in the field for the 97 free-ranging pigs according to the study sites are detailed in Table 3. To analyze the concordance between serological and molecular data obtained from free-ranging pigs, a Venn diagram was built using a freely available online software (http://bioinformatics.psb.ugent.be/webtools/Venn). https://doi.org/10.1371/journal.pntd.0010036.g006, https://doi.org/10.1371/journal.pntd.0010036.t004. Visualization, e0010036. -, Welburn S, Coleman PG, Maudlin I, Fvre E, Odiit M, Eisler MC. 2012 Oct 1;5:217. doi: 10.1186/1756-3305-5-217. This resulted in many p-values presented in supporting information. The https:// ensures that you are connecting to the Economic losses due to AAT are measured in billions of dollars [5,6]. We highlight the lack of tools to prove or exclude with certainty the presence of T. b. gambiense. CATT-B was performed for each pig and CATT-P was performed for CATT-B positive pigs as described above. 2011 Jan 18;5(1):e941. The LiTat 1.3 gene was already observed in T. b. brucei isolated from pigs in Cte dIvoire [68] and positive TL results were observed in cattle in rhodesiense HAT and AAT endemic region in Uganda [56]. Prevalence of Trypanosome Species in Cattle Near Ruma National Park, Lambwe Valley, Kenya: An Update From the Historical Focus for African Trypanosomosis. The SD Bioline HAT rapid diagnostic tests (RDT) [55], another serological test initially developed for HAT diagnosis, also showed a lack of T. b. gambiense specificity when used in animals [41,56,57]. Writing review & editing, Affiliations BCT = Buffy Coat Technique, CATT-B = Card Agglutination Test for Trypanosomiasis performed on blood, TG = Trafla Gottron. Only one strain from Gozy was clearly positive after a single round TgsGP PCR (S1 Fig). For the animal survey, considering T. brucei s.l.

2003;31:9971013. von Wissmann B, Machila N, Picozzi K, Fvre EM, deC Bronsvoort BM, Handel IG, Welburn SC. Investigation, However, potential animal reservoirs of Trypanosoma brucei gambiense (T. b. gambiense), the main agent of HAT, may compromise this ambitious objective. eCollection 2022. A total of 22 strains were isolated, stabilized and purified (Table 4). Writing review & editing, Roles Writing original draft, Affiliations and T. congolense actively circulated in free-ranging pigs. First, we cannot exclude a contamination during amplification workflow even if all experiments were validated by negative controls. https://doi.org/10.1371/journal.pntd.0010036.t001. Methodology, Roles During a study conducted in this area, we scrutinized the trypanosomes circulating in pigs and people sharing the same environment using serological, immunological, parasitological and molecular tools. The PCR reactions were carried out in a thermocycler (Eppendorf Mastercycler nexus) in 25 l final volume, containing 1 X Qiagen HotStarTaq Master Mix, 20 pmol of each primer and 5 l of DNA sample. Apparent lack of a domestic animal reservoir in Gambiense sleeping sickness in northwest Uganda. Writing original draft, Affiliation DNA from the isolated strains was extracted according to the same process, after having re-suspended the parasite pellets in 200 L PBS. Would you like email updates of new search results? Nineteen pigs were infected with Trypanosoma brucei s. l. (10.1%) of which eight were identified as carrying the human infective sub-species Trypanosoma brucei rhodesiense (4.8%). Laboratoire de Biodiversit et Gestion des Ecosystmes Tropicaux, Unit de Recherche en Gntique et Epidmiologie Molculaire, UFR Environnement, Universit Jean Lorougnon Gud, Daloa, Cte dIvoire, Significant differences were observed between the study sites for LiTat 1.5 and LiTat 1.6 mainly due the high positivity rates observed in Gozy and PK14 (between 70 and 80%), the low positivity rate of LiTat 1.5 and LiTat 1.6 in Gatifla and the absence of LiTat 1.5 in TG. The nested TgsGP PCR gave strongly positive bands for five additional strains from Gozy (2), PK 14 (2) and TG (1) (S2 Fig). eCollection 2021. A long-term residual human reservoir could also explain these infections as already suspected, especially in the neighboring foci of Sinfra and Bonon [33]. The corresponding free-ranging pig was positive to TBR PCR (on both BC and isolated strain) and to LiTat 1.5 and 1.6 TL (not 1.3). Project administration, Formal analysis, All positive CATT-P at a dilution of 1:4 or higher underwent parasitological investigations by direct examination of a lymph node aspirate and/or mini-anion exchange centrifugation technique (mAECT, provided by Projet de Recherches Cliniques sur la Trypanosomiase, Daloa, Cte dIvoire) [22] performed with 350 L of buffy coat (BC) as previously described [23]. Estimated number of pigs in the East African countries of Burundi, Ethiopia, Kenya,, Map of Tanzania, showing districts where pigs were sampled (courtesy of H. K., MeSH Competing interests: The authors have declared that no competing interests exist. The corresponding PCR products could not be sequenced due to a limited amount of product. Only one sample out of 55 TBR PCR positive samples (1.8%) did not exhibit a positive serology and 72% (40/55) of the TBR PCR positive samples were positive for both CATT-B and TL tests. Formal analysis, Validation, Results were very heterogeneous between the study sites but no significant differences were observed (Fig 3). Nakayima J, Nakao R, Alhassan A, Mahama C, Afakye K, Sugimoto C. Parasit Vectors. The black stars represent the individual localization of the HAT cases diagnosed between 2000 and 2012.

World Health Organization now targets the interruption of transmission for 2030. However, 19 (86%) and 9 (41%) strains were positive to TBR and TCF, respectively.

For CATT-B positive persons, 5mL of blood were collected in heparinized tubes and a twofold plasma dilution series in CATT buffer was tested to assess the highest dilution that was still positive on plasma (CATT-P). The important heterogeneity observed regarding the TL profile results according to the study sites suggested the likely circulation of different T. brucei s.l. Epub 2018 Sep 1. Only one strain was clearly positive for both the single round and the nested PCR. These were immunosuppressed with cyclophosphamide (300 mg/kg of Endoxan) administered before inoculation and then every 5 days. The site is secure. Visualization, In Cte dIvoire, HAT elimination as a public health problem is being achieved [2,10].

A deworming treatment (Bolumisol, Laprovet) was provided for free to all pigs sampled. Over 70% of free-ranging pigs were positive for CATT and parasitological investigations and 2743% were positive to trypanolysis depending on the antigen used. An integrated management approach, based on the One health concept [7], is thus recommended for a sustainable elimination of African trypanosomiases [8,9]. It is the case in the Vavoua historical focus in Cte dIvoire where HAT cases were still diagnosed in the early 2010s. No HAT cases were detected among 345 people tested. https://doi.org/10.1371/journal.pntd.0010036.t002. On the other hand, negative samples for the single round PCR were unambiguously positive for the nested PCR. Unable to load your collection due to an error, Unable to load your delegates due to an error, Estimated number of pigs in the East African countries of Burundi, Ethiopia, Kenya, Madagascar, Malawi, Mauritius, Mozambique, Reunion, Rwanda, Seychelles, Somalia, Uganda, Tanzania, Zambia, and Zimbabwe, 1961 2011, according to FAOSTAT online database [.

Methodology, The authors also thank Gervais Douvin for helpful advice in writing the manuscript. Investigation, A total of 167 pigs were tested and the complete database is given in S1 Table. Another limitation concerns the lack of data collection on both the human and pig population sizes, number of pig breeders and number of pigs per breeder in each study sites. More than 90% of the mAECT performed on CATT-B positive pigs were positive. https://doi.org/10.1371/journal.pntd.0010036.s005. ** PCR TBR (TBR1-2 primers) performed on blood samples. Validation, Affiliation This site needs JavaScript to work properly. Such contamination may have occurred in any of the laboratories that routinely work with T. b. gambiense DNA, mainly when using nested PCR to increase sensitivity. Formal analysis, In Cte dIvoire, One Health approaches are being conducted to adapt strategies for reaching HAT sustainable elimination. The CATT-B test showed high positivity rates up to 100% in three free-ranging pig sites. Conclusion: This sampling consisted on 70 pigs in pen in PK5, PK8, PK11 and PK14; and 97 free-ranging ones in CIFCI, Dema, Gatifla, Gozy PK14, and TG (Table 2). An epidemic situation broke out in the 1970s. We acknowledge all the technicians from the HAT teams of Institut Pierre Richet of Bouak, University Jean Lorougnon Gud of Daloa, Projet de Recherches Cliniques sur la Trypanosomiase of Daloa and HAT National Elimination Program of Cte dIvoire. Only four cases were still passively diagnosed in the early 2000s. These results show that in Tanzania domestic pigs may act as a significant reservoir for animal trypanosomiasis including the cattle pathogens T. vivax and T. congolense, the pig pathogen T. simiae, and provide a significant reservoir for T. b. rhodesiense, the causative agent of acute Rhodesian sleeping sickness. Trypanosome isolation in the field was performed by intraperitoneal inoculation of 0.5 mL whole blood from each pig positive to BCT and/or mAECT, in two Naval Medical Research Institute (NMRI) mice (produced in CIRDES, Bobo-Dioulasso, Burkina-Faso, from paternal strains purchased from Charles River laboratories, France). Positive results with both PCR were observed for 8 strains (36%) suggesting that mixed infections are common.

Prior to field survey, objectives of the study were explained to the inhabitants and local authorities of each village. Kalayou S, Okal MN, Odhiambo PO, Mathenge K, Gamba DO, Kariuki E, McOdimba F, Masiga D. Front Vet Sci. Conceptualization, One objective was to evaluate the possible human contamination from a pig transmission cycle in a historical HAT focus. Regarding domestic animals, it depends on the population customs, the animal species raised and the husbandry methods. https://doi.org/10.1371/journal.pntd.0010036, Editor: Joshua B. Benoit, University of Cincinnati, UNITED STATES, Received: April 9, 2021; Accepted: November 28, 2021; Published: December 22, 2021. In a recent study targeting TgsGP to detect T. b. gambiense in tsetse flies in a very low HAT prevalence area in Uganda, the TgsGP primers were suspected to cross-react with DNA from an unidentified source after sequencing the DNA products [64]. The vegetation was initially characterized by mesophilic forest with savannah inclusions mainly in the north. This research was part of the Programme Jeune Equipe Associe lIRD (JEAI ERHATryp) financed by the IRD and coordinated by MK. This study once more highlights the need for technical improvements in exploring the epidemiological role of animals in HAT. Investigation, However, the last two cases that were reported in 2011 and 2012 [18] raised the question of the infection origin. Finelle P. African animal trypanosomiasis. Copyright: 2021 Traor et al. Validation,

Disclaimer, National Library of Medicine Kasozi KI, Zirintunda G, Ssempijja F, Buyinza B, Alzahrani KJ, Matama K, Nakimbugwe HN, Alkazmi L, Onanyang D, Bogere P, Ochieng JJ, Islam S, Matovu W, Nalumenya DP, Batiha GE, Osuwat LO, Abdelhamid M, Shen T, Omadang L, Welburn SC. Methodology, doi: 10.1016/S0305-750X(03)00043-3. 1984;2:2330. No, Is the Subject Area "Serology" applicable to this article? In the absence of accurate field serological diagnostic tests for African trypanosomiases in domestic animals and taking into account the limited sensitivity of BCT, we also used the CATT and mAECT following the same procedures as HAT diagnosis, to increase the possibility of both T. b. gambiense and animal trypanosomes detection. MKN was supported for his PhD by a grant from Institut de Recherche pour le Dveloppement (IRD), Allocation de Recherche pour une Thse au Sud. The difference was remarkable in PK14 where 9 out of the 10 pigs BCT-negative were mAECT-positive.

Mix husbandry was only practiced in PK14 and we included an equivalent number of pigs in pen and free-ranging pigs. 2022 Feb 19;15(1):64. doi: 10.1186/s13071-022-05190-1. These results stress the need for an efficient molecular toolbox to easily detect and identify T. b. gambiense in any animal it may infect. Further studies on a larger and more exhaustive sample would allow a more accurate evaluation of the role of pigs in African trypanosomiases epidemiology. Software, It is interesting to stress that for the six TgsGP-positive strains, the three tests performed on blood and plasma gave positive results. Data Availability: All relevant data are within the manuscript and its Supporting Information files.

strains containing different VAT and mixed infections with several strains in single pigs. Investigation, mAECT was significantly more sensitive than BCT (p 0,001) that was only positive for 17 out of the same 57 pigs (30%). Investigation, Acta Trop. Both human and animal plasma samples were processed with the immune trypanolysis test (TL) using cloned populations of T. b. gambiense variant antigen type (VATs) LiTat 1.3, LiTat 1.5 and LiTat 1.6 as previously described [33]. Animal reservoirs can involve wild animals with a very important diversity of species [65,6971]. When parasitaemia reached 32.106 parasites/mL, blood was collected by cardiac puncture of mice as previously described [26] and parasites were separated from mice blood using mAECT.

Project administration, Conceptualization, Rome: FAO; 1983. pp. PLoS Negl Trop Dis. https://doi.org/10.1371/journal.pntd.0010036.s006. A review on the diagnosis of animal trypanosomoses. Project administration, official website and that any information you provide is encrypted Free-ranging pigs obviously represent a potential HAT risk for humans, but may also constitute a potential source of contamination for other domestic animals as zebu cattle that are sensitive to trypanosome infections [74]. TCS and TgsGP single round PCR were negative for all BC tested. For TL results (Fig 4), the global positivity rate for LiTat 1.6 was the highest (43%) ranging from 16 to 80% according to the study site, followed by LiTat 1.5 (41%) ranging from 0 to 80% and LiTat 1.3 (27%) ranging from 8 to 50%. -, Machila N, Wanyangu SW, McDermott J, Welburn SC, Maudlin I, Eisler MC. here. A systematic review and meta-analysis of small ruminant and porcine trypanosomiasis prevalence in sub-Saharan Africa (1986 to 2018). CATT-B = Card Agglutination Test for Trypanosomiasis performed on blood, * For TL (trypanolysis) data, LiTat 1.3 and 1.5 (considered as T. b. gambiense specific) results were fused into one dataset to simplify the visualization and interpretation of the diagram. Bookshelf 145.

M = molecular weight marker (100 bp DNA ladder) Sample 2 (pig 102) = positive with a clearly positive band Samples 1 (pig 82) and 3 (pig 86) = positive with a low intensity band C1+ and C2+ = positive PCR controls C- = negative PCR control. We focused on CATT-B, TL and TBR PCR data obtained from the 97 free-ranging pigs (Fig 6). No sample other than those for routine screening and diagnostic procedures was collected. Clipboard, Search History, and several other advanced features are temporarily unavailable.

Investigation, Yes Intertryp, IRD, Cirad, Universit de Montpellier, Montpellier, France, Roles Epub 2011 Dec 19. We compared the detection power for the different techniques and in the different situations (sites, husbandry method) with Fishers exact tests when possible under the R commander package (rcmdr) [35,36] for R [37], or with 100,000 randomizations in R when necessary, except for CATT-P for which we used a Krukal-Wallis test with rcmdr. 2021 Nov 1;9:720536. doi: 10.3389/fcell.2021.720536. Trends Parasitol. For the animal survey, the buffy coat technique was also used as well as the PCR using Trypanosoma species specific, including the T. b. gambiense TgsGP detection using single round and nested PCRs, performed from animal blood samples and from strains isolated from subjects positive for parasitological investigations. The PCR results obtained for the 97 free-ranging pigs are given in Fig 3. Intertryp, IRD, Cirad, Universit de Montpellier, Montpellier, France.

In this study, we also used the CATT and mAECT initially adapted for HAT diagnosis, to increase the possibility of T. b. gambiense detection.