Trypanosome Infection Research Articles

Impact of trypanosomiasis on male camel infertility

IntroductionBlood parasitism is a significant clinical disease that silently undermines the livestock industry, particularly affecting camels. This study aimed to assess the prevalence of Trypanosoma evansi in Arabian camels (Camelus dromedarius) and its impact on infertility by examining serum protein fractions, lipids, reproductive indices, and the expression of heat shock protein (HSP70) during breeding season.MethodsA total of 107 male post-pubertal camels, aged between 5 and 10 years, were collected randomly from slaughtering house in Assiut Governorate, Egypt.ResultsMicroscopic and serological examinations revealed that 23.4% (25/107) of the camels were infected with T. evansi. Infected camels exhibited a highly significant increase in total serum protein. The assessment of dyslipidemia, measure as binary variables for lipid profiles (cholesterol, triglycerides, HDL, and LDL), indicated a nonsignificant increase in risk of dyslipidemia in infected camels compared to healthy camels. Proteomic analysis identified four major protein fractions in the infected camels compared to healthy camels with molecular weights of 181.72, 87.59, 30.5, and 19.5 kDa using SDS electrophoresis. Testicular tissue of the infected camels showed degeneration and necrotic changes in seminiferous tubules and interstitial tissue, along with edema and congestion. There was a significant reduction in the diameter of seminiferous tubules and germinal epithelium height. A marked reduction in testosterone levels and a high expression of HSP70 in spermatogonia, spermatocytes, Sertoli cells, and Leydig cells were observed.DiscussionConsequently, a combination of physiological and hormonal analyses may serve as a reliable indicator of Trypanosoma infection.

Comparative evaluation of rapid diagnostic test and PCR-based diagnostic assay for identification of trypanosomes in cattle of Apac and Kiryandongo districts, Uganda: A cross sectional study

BackgroundDiagnostic tests and knowledge of their diagnostic accuracies are important for animal trypanosomosis surveillance and treatment.Methodology: A cross-sectional study was conducted in November 2021 to compare the performance of rapid diagnostic test (RDT) and PCR-based assay for the detection of trypanosome infections. Random sampling and probability proportional to size sampling were used to sample study households and animals from the sampled household respectively. Animals were screened for the presence of trypanosomes using both tests. A total of 100 cattle (52 from Apac and 48 from Kiryandongo districts) were included in the study. The percentage of positive tests, sensitivity, and specificity of the tests using mini PCR as a reference were computed. Cohen’s kappa statistics was computed to assess how well the rapid diagnostic test agrees with the mini PCR. McNemar’s statistic assessed if the proportion of positive tests identified by mini PCR significantly differed from that identified by the rapid diagnostic test.ResultsThe mini PCR test detected 31.2% Trypanosome spp positive samples in Kiryandongo while it detected only 5.7% positive samples in Apac district. The rapid diagnostic test (RDT) analysis detected 91.6% Trypanosome spp positive samples in Kiryandongo district and only 46.1% in Apac district. T. congolense was the most common Trypanosoma species identified in Apac (36.5%) and Kiryandongo (47.9%) by the RDT. The percentage of each of the Trypanosome species (T.vivax, T.congolense, and mixed infection of T.vivax and T. congolense) detected by RDT differed significantly (p < 0.001) between the two districts. The RDT had a high sensitivity of 94.4% (95% Confidence Interval (CI): 72.7–99.9) but a very low specificity of 36.6% (95% CI: 26.2–48.0). The kappa test showed evidence of only a slight agreement (kappa = 0.1547, Accuracy = 0.48 (95% Confidence Interval (CI): 0.379,0.5822) between the two tests. The observed agreement between the tests was 47% while the expected agreement was 37.84%.ConclusionThis study found high sensitivity but low specificity of RDT using mini PCR as a reference.

High prevalence of &lt;em&gt;Trypanosoma&lt;/em&gt; infection in Iberian green frogs (&lt;em&gt;Pelophylax perezi&lt;/em&gt;) and evidence of a negative relationship between blood parasites and two indices of frog body condition

Trypanosoma commonly parasitizes anuran hosts but very few studies have investigated ecological relationships in multiparasitized amphibians. We analysed a sample of 29 adult Iberian green frogs (Pelophylax perezi) from a monitored population in central Spain and found that 28 of these individuals (96.5%) were infected with blood parasites. The protozoa genera Lankesterella (Apicomplexa: Eimeriorina) (72.4%) and Trypanosoma (Euglenozoa: Trypanosomatida) (69%) had the highest prevalence, followed by an intraerythrocytic bacteria of the genus Aegyptianella (Pseudomonadota: Rickettsiales) (31.0%). We also report an infection by hematic microfilariae (Nematoda: Spirurida) (6.8%), which to our knowledge represents the first documented case in Iberian amphibians. Infections with more than one parasite type occurred in 62.1% of the frogs. A multimodel inference approach indicated that the infection intensities of Trypanosoma and Aegyptianella were the most important predictors, both negatively affecting the body condition of the frogs. Furthermore, the number of leeches that frogs had when captured showed a strong positive association with Trypanosoma infection intensity. This suggests that leeches act as primary vectors for Trypanosoma. Our results revealed a high taxonomic diversity of blood parasites in green frogs, thus contributing to expand our knowledge of the biodiversity of Mediterranean wetlands and highlighted the potential negative effects of certain infections on the fitness of these amphibian hosts.

Haemoparasite infection risk in multi-host avian system: an integrated analysis.

Avian blood parasites play a crucial role in wildlife health and ecosystem dynamics, exhibiting heterogeneous spatial distribution influenced by various factors. Although factors underlying heterogeneity in infection with blood parasites have been explored in many avian hosts, their importance in the context of host species and the parasite taxon remains poorly understood, particularly in cohabiting host species. Using next-generation sequencing for parasite screening, we investigate the association between Haemoproteus, Plasmodium and Trypanosoma infections in relation to individual parameters, host densities and landscape features in 3 cavity-nesting passerines: great tit (Parus major), blue tit (Cyanistes caeruleus) and collared flycatcher (Ficedula albicollis) in a highly fragmented forest habitat. Overall, Haemoproteus infections predominated, followed by Plasmodium and Trypanosoma, with great tits most and collared flycatchers least parasitized. There were no common patterns across host species in the probability of infection with locally transmitted parasites from each genus. Specifically, in all cases, the effect of particular parameters, if present, was observed only in 1 host species. Body condition influenced Haemoproteus and Plasmodium infections differently in tits. Host density, whether own species or all pooled, explained Haemoproteus infections in great tits and collared flycatchers, and Plasmodium in great tits. Landscape metrics, such as moisture index and distance to coast edge and pastures, affected infection probability in specific host–parasite combinations. Relative risk maps revealed infection risk gradients, but spatial variation repeatability over time was low. Our study highlights the complex dynamics of avian blood parasites in multi-host systems, shedding light on host–parasite interactions in natural ecosystems.

Spiroplasma endosymbiont reduction of host lipid synthesis and Stomoxyn-like peptide contribute to trypanosome resistance in the tsetse fly Glossina fuscipes.

Tsetse flies (Glossina spp.) vector African trypanosomes that cause devastating diseases in humans and domestic animals. Within the Glossina genus, species in the Palpalis subgroup exhibit greater resistance to trypanosome infections compared to those in the Morsitans subgroup. Varying microbiota composition and species-specific genetic traits can significantly influence the efficiency of parasite transmission. Notably, infections with the endosymbiotic bacterium Spiroplasma have been documented in several Palpalis subgroup species, including Glossina fuscipes fuscipes (Gff). While Spiroplasma infections in Gff are known to hinder trypanosome transmission, the underlying mechanisms remain unknown. To investigate Spiroplasma-mediated factors affecting Gff vector competence, we conducted high-throughput RNA sequencing of the midgut tissue along with functional assays. Our findings reveal elevated oxidative stress in the midgut environment in the presence of Spiroplasma, evidenced by increased expression of nitric oxide synthase, which catalyzes the production of trypanocidal nitric oxide. Additionally, we observed impaired lipid biosynthesis leading to a reduction of this important class of nutrients essential for parasite and host physiologies. In contrast, trypanosome infections in Gff's midgut significantly upregulated various immunity-related genes, including a small peptide, Stomoxyn-like, homologous to Stomoxyns first discovered in the stable fly Stomoxys calcitrans. We observed that the Stomoxyn-like locus is exclusive to the genomes of Palpalis subgroup tsetse species. GffStomoxyn is constitutively expressed in the cardia (proventriculus) and synthetic GffStomoxyn exhibits potent activity against Escherichia coli and bloodstream form of Trypanosoma brucei parasites, while showing no effect against insect stage procyclic forms or tsetse's commensal endosymbiont Sodalis in vitro. Reducing GffStomoxyn levels significantly increased trypanosome infection prevalence, indicating its potential trypanocidal role in vivo. Collectively, our results suggest that the enhanced resistance to trypanosomes observed in Spiroplasma-infected Gff may be due to the reduced lipid availability necessary for parasite metabolic maintenance. Furthermore, GffStomoxyn could play a crucial role in the initial immune response(s) against mammalian parasites early in the infection process in the midgut and prevent gut colonization. We discuss the molecular characteristics of GffStomoxyn, its spatial and temporal expression regulation and its microbicidal activity against Trypanosome parasites. Our findings reinforce the nutritional influences of microbiota on host physiology and host-pathogen dynamics.

Diversity of trypanosomes in tsetse fly caught in two “silent” sleeping sickness foci of Bafia and the Manoka Island in Cameroon

BackgroundSurveillance of “silent” human African Trypanosomiasis (HAT) foci is important for the achievement of the World Health Organization (WHO) goal of interrupting the transmission of this disease by 2030. It is in this context that this study was carried out to determine the trypanosome species circulating in the “silent” HAT foci of Bafia and the Manoka island in Cameroon. MethodsIn the Bafia and Manoka HAT foci, georeferenced pyramidal traps were used to trap tsetse flies. After DNA extraction from each whole fly, molecular tools were used to detect different trypanosome species as well as the origin of tsetse fly blood meals. Geographical information system was used to map the trypanosome infections and entomological data and to localize areas at high risk for trypanosome transmission. ResultsFor this study, 1683 tsetse flies were caught and the relative apparent densities was 2.96: 0.03 in the Bafia HAT focus and 5.23 in the Manoka island. For the molecular identification of trypanosomes, 708 non-teneral tsetse flies (8 from Bafia and 700 from Manoka) were randomly selected. The overall trypanosome infection rate was 7.34 % with no infection in the Bafia HAT focus. Among the analysed flies, 4.57 % had trypanosomes of the subgenus Trypanozoon while 4.1 % and 1.13 % had respectively T. congolense and T. vivax. The most common mixed infections were the combination of trypanosomes of the subgenus Trypanozoon and T. congolense. Of the 708 tsetse flies analysed, 134 (18.93 %) tsetse flies were found with residual blood meals, 94 % and 6 % were respectively from humans and dogs. The trapping sites of Plateau, Sandje and Hospital appeared as the areas where contact with tsetse flies is most common. ConclusionThis study revealed a discrepancy in the abundance tsetse flies as well as the trypanosome infection rates in tsetse of the two “silent” HAT foci of Cameroon. The detection of different trypanosome species in tsetse from the Manoka Island highlights their transmission. The high percentage of human blood meals in tsetse flies indicates an important contact between tsetse flies and human; emphasizing the risk of trypanosome transmission to human in this island.

Agonistic song rate positively correlates with male breeding success and avian malaria infection in Acrocephalus paludicola (Aquatic Warbler), a promiscuous songbird with female-only parental care

Abstract The link between male song and reproductive success has been explored at length in socially monogamous birds, but results were inconsistent and under-represented socially non-monogamous species with uniparental care. Here, we evaluated whether metrics of male song in Acrocephalus paludicola (Aquatic Warbler), a promiscuous songbird with female-only care and large repertoires, are associated with fitness- and quality-related traits. We showed that the number of 9-day-old nestlings fathered and probability of avian malaria infection increased with the rate of agonistic song, produced in male–male interactions, and that male-scaled mass index (proxy for energy reserves) positively correlated with the repertoire size. The male breeding success was not explained by repertoire size and song duty cycle. None of the song variables clearly explained the male return rate or change in the tarsus length (proxy for structural body size) and scaled mass index of the fathered chicks between days 2 and 9 post-hatch. No other relationships between the song characteristics and the inbreeding coefficient, scaled mass index, wing-length and Trypanosoma infection status of the males were supported, and Plasmodium infection was unrelated to the song duty cycle and repertoire size. We conclude that in male A. paludicola the repertoire size could be a signal of early developmental stress or current body condition, and the agonistic song rate could be shaped by sexual selection and signal to females a genetic makeup that enhances survival upon infection by Plasmodium.

Tsetse fly density and trypanosoma infection rate in Bedele and Dabo Hana districts of Buno Bedele Zone, Southwest Ethiopia

BackgroundTrypanosomiasis is an infectious disease caused by parasitic protozoa of the genus Trypanosome and primarily transmitted by tsetse flies. This study aimed to determine the density of tsetse flies and the rate of trypanosome infection in the Bedele and Dabo Hana districts of the Buno Bedele Zone in Ethiopia.ResultsA cross-sectional study was conducted from January to February 2023 to catch tsetse flies, determine tsetse density, and estimate the trypanosome infection rate. We used 100 traps (40 NGU, 30 pyramidal, and 30 biconical) to catch the flies. The following standard procedures were followed to identify the specific trypanosome species in the collected tsetse flies: The flies were dissected, and the salivary glands were removed. We placed the salivary glands in a drop of saline solution on a microscope slide. A coverslip was placed over the salivary glands, the slide was examined under a microscope, and the trypanosomes were identified based on their morphology. A total of 3,740 tsetse flies were captured from 100 traps, resulting in an overall apparent density of 18.7 flies per trap per day. Within the study area, only one species of tsetse fly, Glossina tachinoides, was identified. Of the 1,320 dissected Glossina tachinoides, 1.82% were found to be infected with trypanosome parasites. Among these infections, 58.33% were attributed to Trypanosoma congolense, while the remaining 41.67% were caused by Trypanosoma brucei. The infection rate of trypanosomes was significantly higher in female tsetse flies (87.5%) as compared to male flies (12.5%). Furthermore, a significantly higher infection rate was observed in flies older than 20 days (83.33%) and in hunger stage 1 flies (58.33%) compared to hunger stages 2, 3, and 4.ConclusionsThis study highlights the necessity of implementing control and suppression measures targeting the vector (tsetse flies) and the parasite (trypanosomes) to effectively manage and prevent pathogenic animal trypanosomiasis.

Trypanosoma Infection in Freshwater Fish and Its Corellation with Water Quality

Trypanosoma Infection in Freshwater Fish and Its Corellation with Water Quality

Symbiotic bacteria Sodalis glossinidius, Spiroplasma sp and Wolbachia do not favour Trypanosoma grayi coexistence in wild population of tsetse flies collected in Bobo-Dioulasso, Burkina Faso.

Tsetse flies, the biological vectors of African trypanosomes, have established symbiotic associations with different bacteria. Their vector competence is suggested to be affected by bacterial endosymbionts. The current study provided the prevalence of three tsetse symbiotic bacteria and trypanosomes in Glossina species from Burkina Faso. A total of 430 tsetse flies were captured using biconical traps in four different collection sites around Bobo-Dioulasso (Bama, Bana, Nasso, and Peni), and their guts were removed. Two hundred tsetse were randomly selected and their guts were screened byPCR for the presence of Sodalis glossinidius, Spiroplasmasp., Wolbachia and trypanosomes. Of the 200 tsetse, 196 (98.0%) were Glossina palpalis gambienseand 4 (2.0%) Glossina tachinoides. The overall symbiont prevalence was 49.0%, 96.5%, and 45.0%, respectively for S. glossinidius, Spiroplasma and Wolbachia. Prevalence varied between sampling locations: S. glossinidius(54.7%, 38.5%, 31.6%, 70.8%); Spiroplasma (100%, 100%, 87.7%, 100%); and Wolbachia(43.4%, 38.5%, 38.6%, 70.8%),respectively in Bama, Bana, Nasso and Peni. Noteworthy, no G. tachhnoideswas infected by S. glossinidius and Wolbachia, but they were all infected by Spiroplasma sp. A total of 196 (98.0 %) harbored at least one endosymbionts. Fifty-five (27.5%) carried single endosymbiont. Trypanosomes were found only in G.p. gambiense, but not G. tachinoides. Trypanosomes were present in flies from all study locations with an overall prevalence of 29.5%. In Bama, Bana, Nasso, and Peni, the trypanosome infection rate was respectively 39.6%, 23.1%, 8.8%, and 37.5%. Remarkably, only Trypanosoma grayi was present. Of all trypanosome-infected flies, 55.9%, 98.3%, and 33.9% hosted S. glossinidius, Spiroplasma sp and Wolbachia, respectively. There was no association between Sodalis, Spiroplasma and trypanosome presence, but there was a negative association with Wolbachia presence. We reported1.9 times likelihood of trypanosome absence when Wolbachia was present. This is the first survey reporting the presence of Trypanosoma grayi in tsetse from Burkina Faso. Tsetse from these localities were highly positive for symbiotic bacteria, more predominantly with Spiroplasma sp. Modifications of symbiotic interactions may pave way for disease control.

Trypanosoma infection and bloodmeal analysis in post-feeding sand flies across Thailand

Phlebotomine sand flies are recognized as a primary vector of Leishmania and are also suspected vectors of Trypanosoma. The transmission cycle of these parasites relies on the distribution of sand fly vectors, parasites, and reservoir animals. This study aimed to detect Leishmania and Trypanosoma DNA and identify the sources of bloodmeals in post-feeding sand flies captured across Thailand. A total of 42,911 field female sand flies were collected from 11 provinces across Thailand using CDC light traps. Among these, 253 post-feeding sand flies were selected for analysis. The predominant species in this study was Sergentomyia khawi (33.60 %). The DNA was extracted from individual female sand flies. Polymerase chain reaction (PCR), specific to the internal transcribed spacer 1 (ITS1) and the small subunit ribosomal RNA (SSU rRNA) gene regions were used to detect the presence of Leishmania and Trypanosoma DNA, respectively. Additionally, cytochrome c oxidase subunit I (COI) gene region was utilized to identify the sources of host bloodmeals. Leishmania DNA was not detected in any specimens. The analysis of SSU rRNA sequences revealed the presence of Trypanosoma DNA (11.46 %, 29/253) in sand fly samples. Among these samples, T. noyesi (1.58 %, 4/253) was identified in Idiophlebotomus longiforceps and Phlebotomus asperulus, Trypanosoma Anura01+02/Frog2 (1.18 %, 3/253) in Se. khawi, and Trypanosoma Anura04/Frog1 (8.70 %, 22/253) in Se. khawi, Se. hivernus and Grossomyia indica. Bloodmeal analysis utilizing the COI gene revealed a diverse range of vertebrate hosts’ blood, including bird, bat, frog and sun skink. Our findings confirm the presence of Trypanosoma DNA and identify the sources of bloodmeals from vertebrate hosts in various sand fly species, suggesting their potential as possible vectors for Trypanosoma in Thailand. Furthermore, our study is the first to provide molecular evidence using the COI gene to identify frogs as a host blood source for sand flies in Thailand. Further studies focusing on the isolation of live parasites in sand flies to confirm vector potential and examining the role of animal reservoirs will enhance our understanding of the host-parasite relationship and enable more efficient control for disease transmission.

Bovine trypanosomosis, vector distribution and infection rate in three districts of Gamo Zone, southwestern Ethiopia

African animal trypanosomosis is one of the main obstacles to the development of livestock and agricultural output in Ethiopia. It usually results in a severe, frequently fatal sickness, and the infected animals were more weakened as the disease progress and become unfit for work. A cross sectional study design was conducted from December 2021 to April 2022 with the aim of estimating the prevalence of trypanosome infection both in Glossina spp. and cattle, and to assess apparent density of Glossina spp. A total of 298 cattle were selected and examined for trypanosome by using buffy coat technique. The overall prevalence of bovine trypanosomosis was 19.1%; and two species of trypanosomes, T. congolense and T. vivax, were identified in the study area. The prevalence of T. congolense and T. vivax were 15.8% and 2.3%, respectively. The prevalence of trypanosomosis was significantly higher in adult animals (OR = 2.7; p < 0.05) than in younger cattle and poor body condition (OR = 3.18; p < 0.05) than medium body condition animals. The mean PCV value of infected animals was 14.3% (13.3–15.4) significantly lower than the non-infected animals 18.5% (17.8–19.2). Glossina pallidipes is the only tsetse species encountered in all the study areas. In total, 2992 flies were caught of which 90.8% belong to G. pallidipes and 9.2% were other biting flies. The overall apparent density of G. pallidipes was 20.1 F/T/D and other biting flies were 2.0 F/T/D. A total of 307 live Glossina pallidipes were dissected. The overall prevalence of Glossina pallidipes infection rate was 9.1% (95% CI = 5.9–12.4). The prevalence of G. pallidipes infection was significantly higher in Kucha district (OR = 3.2, χ2 = 2.6, p < 0.05) than the other two districts, Daramalo and Arba Minch Zuria. Also it was significantly higher in flies trapped from riverine forest areas (OR = 5.5, χ2 = 2.86, p < 0.05). Therefore, to reduce the impact of trypanosomosis and Glossina, vector control and treating infected cattle with prophylactic or chemotherapeutic drugs and active community participation can play a key role.

Iatrogenic transmission of Trypanosoma evansi infection in camels and its consequences

Trypanosoma evansi infection has started to become a wide spread phenomena around the camel-rearing areas of North Africa and the Middle East. The disease caused by trypanosomes is locally known as “Surra” and it can seriously impact not only the health of domestic animals but the local economy as well. After taking over the management of a farm containing approximately 700 camels, it was found that a large number were suffering from trypanosome infection and it was of the utmost importance to find the source of this infection. An extensive dive into the records and observations were initially made to identify the infected population. Under closer inspection it was found that the infection was limited mostly to female individuals that had undergone extended reproductive analysis or treatment. Blood samples were taken from each of the individuals for buffy coat test and blood smears. Among the total number of tested camels (n = 590), almost 40% were infected with trypanosomes. The number and percentage of infection correlate with the number of fertility and pregnancy treatments that the camels had undergone. The most severely infected group, underwent between 17 and 20 instances of treatment or tests, had an infection rate of almost 90%. The devastating effect of trypanosomiasis on camel pregnancy and birth were also verified with 61% of all abortions and 82% of all neonatal deaths coming from trypanosome infected individuals. These results clearly demonstrate how damaging iatrogenic infections of T. evansi can be and how simply they could have been prevented.

A narrative review on trypanosomiasis and its effect on food production

Trypanosomiasis is an endemic parasitic disease affecting both humans and animal with a severe negative impact on food production in almost all parts of the world. This review seeks to summarize the history of trypanosomiasis and examine the prevalence of trypanosome infection in animals and its effects on food production. A narrative review was conducted on the history of trypanosomiasis. The literature search was conducted on different databases, and selected articles were screened, data extracted, and analyzed. It is believed that trypanosomiasis has been in existence for several decades dating as far as about 2500 BC during the era of the Egyptian kingdom. Africa was found to have the most common cases of animal trypanosomiasis, with 36 out of 40 articles reporting cases. Cattle among other mammals are the most studied animals and they are the most affected. Hence, milk and meat production are greatly affected by trypanosomiasis. The age of animals and the feed provided to animals also played a role in the prevalence and distribution of the pathogen. This review reveals a wide geographical distribution and diverse host range of trypanosome species. The study also highlights the severity of trypanosomiasis and its impact on food production.

A Nanobody/Monoclonal Antibody "hybrid" sandwich technology offers an improved immunoassay strategy for detection of African trypanosome infections.

The scarcity of reliable devices for diagnosis of Animal African trypanosomiasis (AAT) presents a limitation to control of the disease. Existing high-sensitivity technologies such as PCR are costly, laborious, time-consuming, complex, and require skilled personnel. Hence, utilisation of most diagnostics for AAT is impracticable in rural areas, where the disease occurs. A more accessible point-of-care test (POCT) capable of detecting cryptic active infection, without relying on expensive equipment, would facilitate AAT detection. In turn, early management, would reduce disease incidence and severity. Today, several ongoing research projects aim at modifying complex immunoassays into POCTs. In this context, we report the development of an antigen (Ag) detection sandwich ELISA prototype for diagnosis of T. congolense infections, which is comprised of nanobody (Nb) and monoclonal antibody (mAb) reagents. The Nb474H used here, originated from a past study. Briefly, the Nb was engineered starting from mRNA of peripheral blood lymphocytes of an alpaca immunized with soluble lysate of Trypanosoma congolense (TC13). T. congolense glycosomal fructose-1,6-bisphosphate aldolase (TcoALD) was discovered as the cognate Ag of Nb474H. In this study, splenocytes were harvested from a mouse immunized with recombinant TcoALD and fused with NS01 cells to generate a hybridoma library. Random screening of the library on TcoALD retrieved a lone binder, designated IgM8A2. Using Nb474H as Ag-capture reagent in combination with the IgM8A2 monoclonal antibody Ag-detection reagent resulted in a tool that effectively detects native TcoALD released during infection by T. congolense parasites. Hitherto, development of POCT for detection of active trypanosome infection is elusive. The Nanobody/Monoclonal Antibody (Nb/mAb) "hybrid" sandwich technology offers prospects for exploration, using the unique specificity of Nb as a key determinant in Ag capturing, while using the versatility of monoclonal Ab to adapt to various detection conditions.

Evaluation of ITS1 rDNA primers for the detection and identification of African trypanosomes in mammalian hosts and tsetse flies.

Although several primers targeted to the internal transcribed-spacer 1 (ITS1) of the ribosomal DNA (rDNA) have been designed to improve the detection of African trypanosomes, no study tried to compare their agreement level and ability to amplify different trypanosome species in tsetse flies and mammals in various epidemiological settings. This study was designed to fill this gap, by targeting tsetse-infested areas of Cameroon. For this, archived DNA samples reporting at-least one trypanosome species with species-specific PCR primers were reviewed. Ten sets of primers targeting different ITS1 rDNA sequences of trypanosomes were selected for assessment using single-round and nested-PCR method. Amplification rates (sensitivity) and agreement level of different ITS1 assays were compared using Cohen's-Kappa and McNemar's x2 statistic. Little agreement level (k = 0.05–0.52) were observed between different ITS1-primers PCRs detection of African trypanosome species despite significant (X2=54.3, p = 0.0001) high amplification rate 91.6 % (339/370). This sensitivity varied from quite low for T. simiae (11.9 %) and T. vivax (27.3 %) to fairly good for T. congolence (51.9 %), Trypanozoon (32.4 %) and T. theileri (40.3 %). Primers set targeting ITS1-A sequence of trypanosome species recorded the highest sensitivity (50.5 %) with fairly good agreement compared to 39.2 % for ITS1-C (k = 0.52), 32.4 % for ITS1-R (k = 0.47), 29.7 % for ITS1-N (k = 0.48) and 23.0 % for ITS1-KIN (k = 0.43) respectively. This study revealed a diversity in the sensitivity of different trypanosome species with different sets of ITS-primers enhancing the need to use the same sets of primers in different bio-ecological settings. The use of nested-PCR instead of single-round PCR enabled improvement of trypanosome infections detection in both tsetse and mammals. Among the sets of ITS1-primers tested, those designed by to amplify ITS1-A can be considered as the most appropriate for the detection of trypanosome infections in mammals and tsetse flies.

Molecular detection of Anaplasma, Babesia, Theileria, and Trypanosoma infection in cattle and buffaloes in India.

The online version contains supplementary material available at 10.1007/s12639-024-01673-3.

Adipose and skin distribution of African trypanosomes in natural animal infections

BackgroundAnimal African trypanosomiasis, which is caused by different species of African trypanosomes, is a deadly disease in livestock. Although African trypanosomes are often described as blood-borne parasites, there have been recent reappraisals of the ability of these parasites to reside in a wide range of tissues. However, the majority of those studies were conducted on non-natural hosts infected with only one species of trypanosome, and it is unclear whether a similar phenomenon occurs during natural animal infections, where multiple species of these parasites may be present.MethodsThe infective trypanosome species in the blood and other tissues (adipose and skin) of a natural host (cows, goats and sheep) were determined using a polymerase chain reaction-based diagnostic.ResultsThe animals were found to harbour multiple species of trypanosomes. Different patterns of distribution were observed within the host tissues; for instance, in some animals, the blood was positive for the DNA of one species of trypanosome and the skin and adipose were positive for the DNA of another species. Moreover, the rate of detection of trypanosome DNA was highest for skin adipose and lowest for the blood.ConclusionsThe findings reported here emphasise the complexity of trypanosome infections in a natural setting, and may indicate different tissue tropisms between the different parasite species. The results also highlight the need to include adipose and skin tissues in future diagnostic and treatment strategies.Graphical

The internal transcribed spacer 1 sequence polymorphism brings updates to tsetse species distribution in the northern Cameroon: Importance in planning efficient vector control.

Vector control remains one of the best strategies to prevent the transmission of trypanosome infections in humans and livestock and, thus, a good way to achieve the elimination of human African trypanosomiasis and animal African trypanosomiasis. A key prerequisite for the success of any vector control strategy is the accurate identification and correct mapping of tsetse species. In this work, we updated the tsetse fly species identification and distribution in many geographical areas in Cameroon. Tsetse flies were captured from six localities in Cameroon, and their species were morphologically identified. Thereafter, DNA was extracted from legs of each tsetse fly and the length polymorphism of internal transcribed spacer-1 (ITS1) region of each fly was investigated using PCR. ITS1 DNA fragments of each tsetse species were sequenced. The sequences obtained were analysed and compared to those available in GenBank. This enabled to confirm/infirm results of the morphologic identification and then, to establish the phylogenetic relationships between tsetse species. Morphologic features allowed to clearly distinguish all the tsetse species captured in the South Region of Cameroon, that is, Glossina palpalis palpalis, G. pallicera, G. caliginea and G. nigrofusca. In the northern area, G. morsitans submorsitans could also be distinguished from G. palpalis palpalis, G. tachinoides and G. fuscipes, but these three later could not be distinguished with routine morphological characters. The ITS1 length polymorphism was high among most of the studied species and allowed to identify the following similar species with a single PCR, that is, G. palpalis palpalis with 241 or 242 bp and G. tachinoides with 221 or 222 bp, G. fuscipes with 236 or 237 bp. We also updated the old distribution of tsetse species in the areas assessed, highlighting the presence of G. palpalis palpalis instead of G. fuscipes in Mbakaou, or in sympatry with G. morsitans submorsitans in Dodeo (northern Cameroon). This study confirms the presence of G. palpalis palpalis in the Adamawa Region of Cameroon. It highlights the limits of using morphological criteria to differentiate some tsetse species. Molecular tools based on the polymorphism of ITS1 of tsetse flies can differentiate tsetse species through a simple PCR before downstream analyses or vector control planning.

The cryo-EM structure of trypanosome 3-methylcrotonyl-CoA carboxylase provides mechanistic and dynamic insights into its enzymatic function

3-Methylcrotonyl-CoA carboxylase (MCC) catalyzes the two-step, biotin-dependent production of 3-methylglutaconyl-CoA, an essential intermediate in leucine catabolism. Given the critical metabolic role of MCC, deficiencies in this enzyme lead to organic aciduria, while its overexpression is linked to tumor development. MCC is a dodecameric enzyme composed of six copies of each α- and β-subunit. We present the cryo-EM structure of the endogenous MCC holoenzyme from Trypanosoma brucei in a non-filamentous state at 2.4Å resolution. Biotin is covalently bound to the biotin carboxyl carrier protein domain of α-subunits and positioned in a non-canonical pocket near the active site of neighboring β-subunit dimers. Moreover, flexibility of key residues at α-subunit interfaces and loops enables pivoting of α-subunit trimers to partly reduce the distance between α- and β-subunit active sites, required for MCC catalysis. Our results provide a structural framework to understand the enzymatic mechanism of eukaryotic MCCs and to assist drug discovery against trypanosome infections.