Epidemiological study on cattle trypanosomiasis and its vectors distributions in the Gambella regional state, southwestern Ethiopia.

African animal trypanosomosis is a parasitic disease that causes significant economic losses in livestock due to anaemia, loss of condition, emaciation, and mortality. It is a key impediment to increased cattle output and productivity in Ethiopia. Cross-sectional entomological and parasitological studies were performed in the Gambella Region state of southwestern Ethiopia to estimate the prevalence of bovine trypanosomosis, apparent fly density, and potential risk factors. Blood samples were taken from 546 cattle for the parasitological study and analyzed using the buffy coat technique and stained with Giemsa. A total of 189 biconical (89) and NGU (100) traps were deployed in the specified districts for the entomological survey. The overall prevalence of trypanosomosis at the animal level was 5.5% (95% CI: 3.86-7.75). Trypanosoma vivax (50.0%), T. congolense (30.0%), T. brucei (20.0%), and no mixed trypanosome species were found. The prevalence of trypanosomosis was significantly (p < 0.05) affected by altitude, body score conditions, age, mean packed cell volume (PCV), and peasant associations, while sex and coat color had no significant effect. According to the entomological survey results, a total of 2303 flies were captured and identified as tsetse (Glossina pallidipes (5.3%)) and G. fuscipes fuscipes (3.3%) and other biting flies (Tabanus (60.1%) and Stomoxys (31.3%)). In the current study, the overall apparent density was 4.1 flies/trap/day. This study shows that trypanosomosis remains a significant cattle disease in the Gambella regional state even during the dry season. Thus, the findings support the necessity to improve vector and parasite control measures in the area.

Detection of anti-Trypanosoma spp. antibodies in cattle from southern Brazil.

Bovine trypanosomosis, caused by Trypanosoma vivax, is a disease that originated in Africa and currently affects cattle in several South American countries, including almost all Brazilian states. Despite the reports on T. vivax infection in southern Brazil, data on its circulation status is currently unavailable. In this study, we aimed to detect anti-Trypanosoma spp. IgG antibodies in cattle from Rio Grande do Sul and suggest areas with T. vivax transmission risk. A total of 691 serum samples from cattle in the intermediate regions of Rio Grande do Sul were analyzed using indirect immunofluorescence assay (IFA). The overall seroprevalence of anti-Trypanosoma antibodies in cattle was 24.6% (170/691). The detection rate ranged from 0-37.3%, with a high prevalence in the intermediate regions of Ijuí (37.3%), Uruguaiana (30.7%), and Passo Fundo (28.9%). Thus, these regions were suggested as possible bovine trypanosomosis risk areas due to the high seroprevalence. This is the first serological study to determine Trypanosoma spp. infection status in cattle from Rio Grande do Sul, providing data on the epidemiology of trypanosomosis in the state.

Study on the prevalence and associated risk factors of bovine trypanosomiasis in Zaba Gazo Woreda, Southern Ethiopia.

In tsetse-infested areas of Ethiopia, African Animal Trypanosomiasis is the most serious threat to livestock productivity. A cross-sectional study was conducted from May 2021 to September 2021 in the Zaba Gazo district to assess the prevalence and associated risk factors of bovine trypanosomiasis through parasitological and hematological techniques. In the current study, the overall prevalence of trypanosomiasis was found to be 21%. Among anticipated risk factors, the origin of animals has shown a statistically significant association (p < 0.05) with the occurrence of bovine trypanosomiasis. Other potential risk factors, such as sex, age, BCS, coat color, and agroecology, were not shown to have a statistically significant relationship (p > 0.05) with trypanosomiasis. The overall mean PCV values for parasitaemic and aparasitaemic cattle were 23.24 ± 3.04SD and 27.37 ± 4.18SD, respectively, thus it was lower in parasitaemic animals than the aparasitaemic animals. A high prevalence of bovine trypanosomiasis in the study area necessitates an integrated parasite and vector control approach to curve the prevalence and effect of the disease.

Novel protein candidates for serodiagnosis of African animal trypanosomosis: Evaluation of the diagnostic potential of lysophospholipase and glycerol kinase from Trypanosoma brucei.

African trypanosomosis, a parasitic disease caused by protozoan parasites transmitted by tsetse flies, affects both humans and animals in sub-Saharan Africa. While the human form (HAT) is now limited to foci, the animal form (AAT) is widespread and affects the majority of sub-Saharan African countries, and constitutes a real obstacle to the development of animal breeding. The control of AAT is hampered by a lack of standardized and easy-to used diagnosis tools. This study aimed to evaluate the diagnostic potential of TbLysoPLA and TbGK proteins from Trypanosoma brucei brucei for AAT serodiagnosis in indirect ELISA using experimental and field sera, individually, in combination, and associated with the BiP C-terminal domain (C25) from T. congolense. These novel proteins were characterized in silico, and their sequence analysis showed strong identities with their orthologs in other trypanosomes (more than 60% for TbLysoPLA and more than 82% for TbGK). TbLysoPLA displays a low homology with cattle (<35%) and Piroplasma (<15%). However, TbGK shares more than 58% with cattle and between 45-55% with Piroplasma. We could identify seven predicted epitopes on TbLysoPLA sequence and 14 potential epitopes on TbGK. Both proteins were recombinantly expressed in Escherichia coli. Their diagnostic potential was evaluated by ELISA with sera from cattle experimentally infected with T. congolense and with T.b. brucei, sera from cattle naturally infected with T. congolense, T. vivax and T.b. brucei. Both proteins used separately had poor diagnostic performance. However, used together with the BiP protein, they showed 60% of sensitivity and between 87-96% of specificity, comparable to reference ELISA tests. In conclusion, we showed that the performance of the protein combinations is much better than the proteins tested individually for the diagnosis of AAT.

In vitro and in vivo effectiveness of disinfectants against Trypanosoma vivax.

Trypanosoma vivax causes bovine trypanosomosis in cattle and resulting in economic losses to farmers. In Brazil, shared contaminated materials is the main transmission pathway. To evaluate the effectiveness of different disinfectants for T. vivax, in vitro and in vivo analyses were performed. At the laboratory, 21 disinfectants were tested. The disinfectants were placed in microtubes containing blood with approximately 1.0 × 106 trypomastigotes of T. vivax. The viability and motile of trypomastigotes after 30 s, one, 10, 15 and 30 min was evaluated by the thick drop method and the efficacy calculated. Disinfectants that showed 100% effectiveness were used in in vivo tests. Thirty calves negative for T. vivax were divided into six groups and were inoculated with disinfectant solutions (46% alcohol, 70% alcohol, or 0.5% iodine) + 1 × 106 trypomastigotes of the protozoa. Blood from each animal was collected at seven, 14 and 21 days after inoculation to verify the viability and presence of the protozoan by Woo, Brener, PCR, and LAMP methods. In the in vitro step, 13 of the 21 disinfected solutions exhibited 100% effectiveness against T. vivax at all evaluation times. In contrast, 70% alcohol and 0.5% iodine solutions exhibited 100% effectiveness in the in vivo tests and can be used to disinfect needles and syringes. The use of disinfectants is a rapid and efficient procedure to disinfect materials utilized in the field and concomitantly could help to reduce the dissemination of T. vivax in the cattle herd in cases of iatrogenic transmission.

Evaluation of five primer sets for molecular detection of Trypanosoma vivax by polymerase chain reaction (PCR) and their implementation for diagnosis in naturally infected ruminants from Venezuela.

Trypanosoma vivax is a protozoan parasite that causes trypanosomosis in ruminants and is widely distributed in tropical areas in the world. The control of this disease depends on the sensitivity and specificity of the diagnostic tests implemented for naturally infected samples, where parasitaemias are usually low. This study aimed to evaluate the analytical sensitivity and specificity of several primers for T. vivax detection in experimental infections and their implementation for the diagnosis of trypanosomosis in naturally infected bovine and ovine samples. Using a T. vivax Venezuelan isolate, five sets of primers were evaluated: TviSL1/2, ITS1CF/BR, TVMF/R, ILO1264/1265, TVWA/B. Additionally, we tested the PCR protocols using different DNA quantities. The best set of primers (ILO1264/1265) was used to detect T. vivax DNA from whole blood and buffy coat samples of 12 sheep (ovine) and 45 cattle (bovine) of small farms from Venezuela, and compared to the micro-haematocrite centrifugation technique (MHCT). The highest sensitivity was 0.0001 ng for ILO1264/1265 and TVWA/B primers. Using 100 ng of DNA extracted from the buffy coat and the ILO1264/1265 primers for trypanosomosis diagnosis from naturally infected samples, yielded 66.7% (8/12) and 35.7% (16/45) positives in ovine and bovine respectively. The percentage of positives samples increased to 83.3% (10/12) and 64.4% (29/45), with 300 ng in the assays. Contrary, using 300 ng of DNA extracted from the whole blood yielded only 50% (6/12) and 28.9% (13/45) of positives samples for T. vivax respectively. MHCT only detected the parasite in bovine samples with 17.8% (8/45) of positives. Based on our results, we recommend the use of the ILO1264/1265 primers and 300 ng of DNA extracted from the buffy coat for epidemiological studies of naturally infected animals. Moreover, detection of the parasite in ovine herds highlights a possible role of this host in the epidemiology of trypanosomosis in Venezuela.

Diminazene resistance in Trypanosoma congolense is not caused by reduced transport capacity but associated with reduced mitochondrial membrane potential.

Trypanosoma congolense is a principal agent causing livestock trypanosomiasis in Africa, costing developing economies billions of dollars and undermining food security. Only the diamidine diminazene and the phenanthridine isometamidium are regularly used, and resistance is widespread but poorly understood. We induced stable diminazene resistance in T. congolense strain IL3000 in vitro. There was no cross-resistance with the phenanthridine drugs, melaminophenyl arsenicals, oxaborole trypanocides, or with diamidine trypanocides, except the close analogs DB829 and DB75. Fluorescence microscopy showed that accumulation of DB75 was inhibited by folate. Uptake of [3 H]-diminazene was slow with low affinity and partly but reciprocally inhibited by folate and by competing diamidines. Expression of T. congolense folate transporters in diminazene-resistant Trypanosoma brucei brucei significantly sensitized the cells to diminazene and DB829, but not to oxaborole AN7973. However, [3 H]-diminazene transport studies, whole-genome sequencing, and RNA-seq found no major changes in diminazene uptake, folate transporter sequence, or expression. Instead, all resistant clones displayed a moderate reduction in the mitochondrial membrane potential Ψm. We conclude that diminazene uptake in T. congolense proceed via multiple low affinity mechanisms including folate transporters; while resistance is associated with a reduction in Ψm it is unclear whether this is the primary cause of the resistance.

Parasitological and clinical-pathological findings in twelve outbreaks of acute trypanosomiasis in dairy cattle in Rio de Janeiro state, Brazil.

In Brazil, infection in cattle was first reported in the state of Pará, in 1944, and the presence of the parasite has already been recorded in several states. The purpose of this study was to report the clinical-pathological aspects of a natural infection by T. vivax in dairy cattle in the state of Rio de Janeiro. Twelve outbreaks of the infection were diagnosed in 11 municipalities from April 2016 to October 2018. All properties had acquired cattle from states where the disease had already been recorded and it was found that needles for oxytocin administration had been shared. These outbreaks were studied by visiting the properties to perform anamnesis, clinical exams and collection of material for laboratory diagnosis. Laboratory diagnosis was performed through parasitological, molecular and histopathological techniques. Animals with confirmed diagnosis for T. vivax showed anemia, lack of appetite, decreased milk production, weight loss, weakness, abortion, diarrhea and neurological signs. The main histological lesions found were meningoencephalitis and lymphohistiocytic myocarditis. In the central nervous system, the lesions were more severe in the brain compared to the spinal cord, being progressively more severe in the rostro-dorsal direction. Also, they were more accentuated in the white matter compared to the gray matter. Due to nonspecific clinical signs, laboratory tests were key for diagnosis. Trypanosomiasis in cattle herds in the state of Rio de Janeiro, Brazil, is of great concern because of its potential to cause economic losses.

Trypanosomosis prevalence in natural field-based infection: insights into systematic review and meta-analysis of studies from 1980 to 2018 on The Gambian ruminants with special emphasis on cattle.

Consistent quantification of trypanosomes, the parasite responsible for African animal trypanosomosis, is important for effective surveillance, control, and eradication strategies. Here, we used a rigorously predefined protocol to search and select eligible publications that utilized either microscopy, serology, or molecular methods to investigate prevalence of trypanosomosis based on the presence of any of three most common Trypanosoma spp. (T. congolense, T. vivax, and T. brucei) in the field-based naturally grazed Gambian cattle, sheep, and goats. To combine results of studies on cattle through meta-analysis, sensitivity and subgroup analyses were carried out with the random effects model, and prevalence estimates of each study with 95% confidence intervals (CI) were presented with a forest plot. All the eligible studies utilized the buffy coat technique (BCT) to detect trypanosomes in the blood samples, while the more sensitive serological and molecular detection methods are yet to be widely exploited. Heterogeneity among the studies on cattle was moderate (I2 = 55%), and the pooled trypanosomosis prevalence based on the BCT was 5.2% (95% CI: 4.0-6.4). Meanwhile, estimated prevalence varied according to the trypanosome detection methods, study locations, types of publication, year, and length of observations. We could not pool the trypanosomosis prevalence in sheep and goats through meta-analysis due to small number of studies. The prevalence estimates based on the BCT ranged from 3.2 to 8.1% in goats and 2.8 to 10.6% in sheep. Even though there seems to be a slight decrease in trypanosomosis prevalence in cattle in one of the Gambian districts, there was no consistent trend across the years. It is thought that the literature search and formatting procedures presented in this study contribute to doing systematic reviews on the investigated subject and can be adapted for similar cases.

A case of bovine trypanosomiasis caused by Trypanosoma theileri in Sicily, Italy.

Despite some researchers reporting clinical signs in cattle associated with Trypanosoma theileri, its role as a pathogen is still unclear. We describe here the isolation of Trypanosoma theileri during a routine laboratory investigation. Mature and immature vital parasitic forms were observed within hematopoietic cell cultures from the bone marrow of one cow for monocyte isolation. The animal was submitted to clinical examination and blood sample counting (CBC). Postmortem analysis included gross and histological examination and PCR in the liver, spleen, brain, lymph nodes, and lungs. PCR and Giemsa staining were used for parasite identification. A second cow belonging to the same farm was positive for Trypanosoma theileri by PCR performed on blood sample. In this case, the postmortem analysis included also testis. Clinical examination showed only a reduction in body weight in both cases. The CBC revealed an increase of lymphocytes and neutrophils while red blood cells were within the normal range. Spleen was slightly increased in volume and the histology revealed a proliferative activity of the white and red pulp. The biomolecular analysis identified the parasite as Trypanosoma theileri and its DNA was detected in the bone marrow, testis, and brain. The unusual finding of parasite in the brain, testis, and bone marrow raises new clinical implication on disease course and also possible sexual transmission.

Molecular identification of bovine trypanosomes in relation to cattle sources in southwest Nigeria.

Bovine trypanosomosis is a problem in the livestock industry in Nigeria. A longitudinal survey of cattle sampled during the wet and dry seasons was conducted from April 2016 to March 2017. Blood samples were collected by random sampling from 745 cattle in southwest Nigeria and screened for trypanosomes by internal transcribed spacer-polymerase chain reaction (ITS-PCR). Cattle positive for Trypanozoon DNA were further screened with the Rode Trypanozoon antigen type (RoTat) 1.2 PCR and Trypanosoma brucei gambiense glycoprotein (TgsGP) genes for T. evansi and T. b. gambiense respectively. Trypanosome DNA was amplified in 23.8% (95%CI: 20.8-26.9) of cattle with significantly higher prevalence in wet season (95%CI: 22.9-30.8) when compared to the dry season (95%CI: 14.3-23.6). A high prevalence was observed in Fulani cattle farms 54.1% (95%CI: 42.78-64.93%) while the prevalence was lower in institutional farms 14.7% (95%CI: 10.10-20.97%). Trypanosoma vivax was the most prevalent trypanosome observed (11.54% (95%CI: 9.44-14.04%)), followed by T. congolense 8.5% (95%CI: 6.67-10.67%) T. b. brucei 4.8% (95%CI: 3.51-6.62%) and T. evansi 1.74% (95%CI: 1.02-2.96%). Mixed infections were observed in 2.8% (95%CI: 1.85-4.27%) of cattle. Seasonal variation revealed a predominance of T. congolense and T. vivax in wet and dry season, respectively. The high prevalence of Trypanosoma species in cattle indicates a need for expanded surveillance for AAT in southwest Nigeria. Migration, settlement patterns, increased marketing and management types were some of the risk factors identified for AAT.

Trypanosoma brucei RRP44 is involved in an early stage of large ribosomal subunit RNA maturation.

Ribosomal RNA precursors undergo a series of structural and chemical modifications to generate matured RNA molecules that will comprise ribosomes. This maturation process involves a large set of accessory proteins as well as ribonucleases, responsible for removal of the external and internal transcribed spacers from the pre-rRNA. Early-diverging eukaryotes belonging to the Kinetoplastida class display several unique characteristics, in particular in terms of RNA synthesis and maturation. These peculiarities include the rRNA biogenesis and the extensive fragmentation of the large ribosomal subunit (LSU) rRNA. The role of specific endo- and exonucleases in the maturation of the unusual rRNA precursor of trypanosomatids remains largely unknown. One of the nucleases involved in rRNA processing is Rrp44, an exosome associated ribonuclease in yeast, which is involved in several metabolic RNA pathways. Here, we investigated the function of Trypanosoma brucei RRP44 orthologue (TbRRP44) in rRNA processing. Our results revealed that TbRRP44 depletion causes unusual polysome profile and accumulation of the complete LSU rRNA precursor, in addition to 5.8S maturation impairment. We also determined the crystal structure of TbRRP44 endonucleolytic domain. Structural comparison with Saccharomyces cerevisiae Rrp44 revealed differences in the catalytic site and substitutions of surface residues, which could provide molecular bases for the lack of interaction of RRP44 with the exosome complex in T. brucei.

Genetic diversity and molecular survey of Trypanosoma (Megatrypanum ) theileri in cattle in Brazil's western Amazon region / Diversidade genética e levantamento molecular de Trypanosoma (Megatrypanum) theileri em bovinos na região da Amazônia Ocidental do Brasil

Abstract Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL.

Resumo Trypanosoma (Megatrypanum) theileri é um protozoário flagelado que infecta ruminantes e apresenta alta diversidade genética. Neste estudo, investigamos as taxas de prevalência deste protozoário com base na hemocultura e no diagnóstico molecular. Os isolados de T . theileri obtidos foram caracterizados pelos marcadores moleculares SSU rDNA e gGAPDH e o diagnóstico molecular foi baseado no gene do tipo Catepsina L (PCR-TthCATL). O PCR-TthCATL e a hemocultura indicaram uma taxa de prevalência total de 8,13% e a sequência derivada do gene Catepsina L denominada IB de T. theileri foi identificada pela primeira vez em bovinos da Amazônia Ocidental, bem como a IF no Brasil. Também descrevemos uma possível nova sequência derivada da PCR-TthCATL em bovinos, designada IL.

Haematology of N'Dama and West African Shorthorn cattle herds under natural Trypanosoma vivax challenge in Ghana.

Background: Animal trypanosomosis is a major cause of economic loss in livestock production in Africa. A suggested control measure is to use breeds with traits of trypanotolerance. The study examines the effect of natural Trypanosoma vivax challenge on haematological parameters in two trypanotolerant cattle [N'Dama and West African Short Horn (WASH)] herds. Methods:T. vivax-specific primers were used to diagnose T. vivax infection in an N'Dama herd at Cape Coast in southern Ghana and a WASH herd at Chegbani in northern Ghana from May to July 2011 in a cross-sectional study. Levels of haematological parameters comprising packed cell volume (PCV), haemoglobin (Hb) concentration and total red blood cell (RBC) and white blood cell (WBC) counts; differential WBC counts (neutrophils, lymphocytes, eosinophils, monocytes and basophils); and RBC indices of mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were determined in blood samples and then compared between infected and uninfected cattle. Results: We found that haematological indices for infected and uninfected animals in both breeds were within the normal range. However, the mean PCV values for T. vivax-infected WASH and N'Dama were lower in infected compared to uninfected animals. The difference was significant ( p< 0.05) in N'Dama but not in WASH. The RBC indices were higher in infected N'Dama compared to infected WASH with a significant difference in total RBC ( p < 0.05). Conclusion: We conclude from our findings that despite the presence of infection by T. vivax, N'Dama and WASH cattle maintained their haematological parameters within acceptable normal ranges, and this underscores the need for routine diagnosis and treatment so that such trypanotolerant cattle do not serve as potential reservoirs of trypanosome parasites.

Genetic diversity and molecular survey of Trypanosoma (Megatrypanum ) theileri in cattle in Brazil's western Amazon region.

Trypanosoma (Megatrypanum) theileri is a flagellated protozoan that infects ruminants and it displays high genetic diversity. In this study, we investigated the prevalence rates of this protozoan based on hemoculture and molecular diagnosis. The isolates of T. theileri thus obtained were characterized by molecular markers SSU rDNA and gGAPDH and molecular diagnosis based on Cathepsin L-like gene (PCR-TthCATL). The PCR-TthCATL and hemoculture indicated an overall prevalence rate of 8.13%, and the CATL derived sequence named IB was identified for the first time in cattle in the western Amazon region, as well as IF in Brazil. We also describe a possible new PCR-TthCATL derived sequence in cattle, designated IL.

A meta-analysis of the prevalence of African animal trypanosomiasis in Nigeria from 1960 to 2017.

BACKGROUND: African animal trypanosomiasis is an economically significant disease that affects the livestock industry in Nigeria. It is caused by several parasites of the genus Trypanosoma. National estimates of the disease prevalence in livestock and tsetse flies are lacking, therefore a systematic review and meta-analysis were performed to understand the trend of the disease prevalence over the years. METHODS: Publications were screened in Web of Science, Ovid MEDLINE, Global Health, EMBASE and PubMed databases. Using four-stage (identification, screening, eligibility and inclusion) process in the PRIMSA checklist, only studies that met the inclusion criteria for AAT and tsetse infections were analysed. Point estimates prevalence and subgroup analyses based on diagnostic techniques in livestock were evaluated at 95% confidence interval (CI). RESULTS: A total of 74 eligible studies published between 1960 and 2017 were selected for meta-analysis. This covers the six geopolitical zones, involving a total of 53,924 animals. The overall prevalence of AAT was 16.1% (95% CI: 12.3-20.3%). Based on diagnostic techniques, the prevalence of AAT in cattle was highest in PCR followed by serology and microscopy while the highest prevalence in pigs was observed with serology. Out of 12,552 tsetse flies examined from 14 eligible studies, an overall prevalence of 17.3% (95% CI: 4.5-36.0%) and subgroup prevalence of 49.7% (95% CI: 30.7-68.8%), 11.5% (95% CI: 6.1-18.5) and 4.5% (95% CI: 1.8-8.8%) in G. morsitans, G. tachinoides and G. palpalis, respectively, were observed using the random effects-model. CONCLUSIONS: The prevalence of trypanosomes in both vectors and animal hosts was high in Nigeria. Therefore, further research on risk factors, seasonal and transhumance effects, vectoral capacity and competence are warranted for an effective control of AAT in Nigeria.

A 3D-QSAR Study on the Antitrypanosomal and Cytotoxic Activities of Steroid Alkaloids by Comparative Molecular Field Analysis.

As part of our research for new leads against human African trypanosomiasis (HAT), we report on a 3D-QSAR study for antitrypanosomal activity and cytotoxicity of aminosteroid-type alkaloids recently isolated from the African medicinal plant Holarrhena africana A. DC. (Apocynaceae), some of which are strong trypanocides against Trypanosoma brucei rhodesiense (Tbr), with low toxicity against mammalian cells. Fully optimized 3D molecular models of seventeen congeneric Holarrhena alkaloids were subjected to a comparative molecular field analysis (CoMFA). CoMFA models were obtained for both, the anti-Tbr and cytotoxic activity data. Model performance was assessed in terms of statistical characteristics (R², Q², and P² for partial least squares (PLS) regression, internal cross-validation (leave-one-out), and external predictions (test set), respectively, as well as the corresponding standard deviation error in prediction (SDEP) and F-values). With R² = 0.99, Q² = 0.83 and P² = 0.79 for anti-Tbr activity and R² = 0.94, Q² = 0.64, P² = 0.59 for cytotoxicity against L6 rat skeletal myoblasts, both models were of good internal and external predictive power. The regression coefficients of the models representing the most prominent steric and electrostatic effects on anti-Tbr and for L6 cytotoxic activity were translated into contour maps and analyzed visually, allowing suggestions for possible modification of the aminosteroids to further increase the antitrypanosomal potency and selectivity. Very interestingly, the 3D-QSAR model established with the Holarrhena alkaloids also applied to the antitrypanosomal activity of two aminocycloartane-type compounds recently isolated by our group from Buxus sempervirens L. (Buxaceae), which indicates that these structurally similar natural products share a common structure⁻activity relationship (SAR) and, possibly, mechanism of action with the Holarrhena steroids. This 3D-QSAR study has thus resulted in plausible structural explanations of the antitrypanosomal activity and selectivity of aminosteroid- and aminocycloartane-type alkaloids as an interesting new class of trypanocides and may represent a starting point for lead optimization.

Integrated cost-benefit analysis of tsetse control and herd productivity to inform control programs for animal African trypanosomiasis.

BACKGROUND: Animal African trypanosomiasis (AAT) and its tsetse vector are responsible for annual losses estimated in billions of US dollars ($). Recent years have seen the implementation of a series of multinational interventions. However, actors of AAT control face complex resource allocation decisions due to the geographical range of AAT, diversity of ecological and livestock systems, and range of control methods available. METHODS: The study presented here integrates an existing tsetse abundance model with a bio-economic herd model that captures local production characteristics as well as heterogeneities in AAT incidence and breed. These models were used to predict the impact of tsetse elimination on the net value of cattle production in the districts of Mambwe, in Zambia, and Faro et Déo in Cameroon. The net value of cattle production under the current situation was used as a baseline, and compared with alternative publicly funded control programmes. In Zambia, the current baseline is AAT control implemented privately by cattle owners (Scenario Z0). In Cameroon, the baseline (Scenario C0) is a small-scale publicly funded tsetse control programme and privately funded control at farm level. The model was run for 10 years, using a discount rate of 5%. RESULTS: Compared to Scenario C0, benefit-cost ratios (BCR) of 4.5 (4.4-4.7) for Scenario C1 (tsetse suppression using insecticide treatment of cattle (ITC) and traps + maintenance with ITC barrier), and 3.8 (3.6-4.0) for Scenario C2 (tsetse suppression using ITC and traps + maintenance with barrier of targets), were estimated in Cameroon. For Zambia, the benefit-cost ratio calculated for Scenarios Z1 (targets, ITC barrier), Z2 (targets, barrier traps), Z3 (aerial spraying, ITC barrier), and Z4 (aerial spraying, barrier traps) were 2.3 (1.8 - 2.7), 2.0 (1.6-2.4), 2.8 (2.3-3.3) and 2.5 (2.0-2.9), respectively. Sensitivity analysis showed that the profitability of the projects is relatively resistant to variations in the costs of the interventions and their technical efficiency. CONCLUSIONS: It is envisioned that the methodologies presented here will be useful for the evaluation and design of existing and future control programmes, ensuring they have tangible benefits in the communities they are targeting.

Isothermal microcalorimetry - A quantitative method to monitor Trypanosoma congolense growth and growth inhibition by trypanocidal drugs in real time.

Trypanosoma congolense is a protozoan parasite that is transmitted by tsetse flies, causing African Animal Trypanosomiasis, also known as Nagana, in sub-Saharan Africa. Nagana is a fatal disease of livestock that causes severe economic losses. Two drugs are available, diminazene and isometamidium, yet successful treatment is jeopardized by drug resistant T. congolense. Isothermal microcalorimetry is a highly sensitive tool that can be used to study growth of the extracellular T. congolense parasites or to study parasite growth inhibition after the addition of antitrypanosomal drugs. Time of drug action and time to kill can be quantified in a simple way by real time heat flow measurements. We established a robust protocol for the microcalorimetric studies of T. congolense and developed mathematical computations in R to calculate different parameters related to growth and the kinetics of drug action. We demonstrate the feasibility and benefit of the method exemplary with the two standard drugs, diminazene aceturate and isometamidium chloride. The method and the mathematical approach can be translated to study other pathogenic or non-pathogenic cells if they are metabolically active and grow under axenic conditions.

Mini-review on CRISPR-Cas9 and its potential applications to help controlling neglected tropical diseases caused by Trypanosomatidae.

The CRISPR-Cas system, which was originally identified as a prokaryotic defense mechanism, is increasingly being used for the functional study of genes. This technology, which is simple, inexpensive and efficient, has aroused a lot of enthusiasm in the scientific community since its discovery, and every month many publications emanate from very different communities reporting on the use of CRISPR-Cas9. Currently, there are no vaccines to control neglected tropical diseases (NTDs) caused by Trypanosomatidae, particularly Human African Trypanosomiasis (HAT) and Animal African Trypanosomoses (AAT), and treatments are cumbersome and sometimes not effective enough. CRISPR-Cas9 has the potential to functionally analyze new target molecules that could be used for therapeutic and vaccine purposes. In this review, after briefly describing CRIPSR-Cas9 history and how it works, different applications on diseases, especially on parasitic diseases, are reviewed. We then focus the review on the use of CRISPR-Cas9 editing on Trypanosomatidae parasites, the causative agents of NTDs, which are still a terrible burden for human populations in tropical regions, and their vectors.