Search Results (17)

Bovine trypanosomiasis, caused by Trypanosoma vivax, affects livestock productivity and is increasingly being reported in South America. This study aimed to detect and characterize Trypanosoma spp. infections, with a focus on T. vivax, in cattle from two distinct agroecological regions of central Argentina: a dairy-producing plain, located in the Espinal ecoregion, and a riparian zone, dedicated to beef production, located in the Delta and Islands of Paraná ecoregion. A total of 220 blood samples were collected from nine cattle farms and analyzed using real-time PCR, melting curve analysis, and the sequencing of 18S rRNA gene fragments. Trypanosoma vivax was detected at low prevalence (2.73%), exclusively in dairy cattle. In contrast, the prevalence of Trypanosoma theileri was much higher (10.91%), and it was found mainly in beef cattle from the riparian region. Phylogenetic analyses confirmed the species identity in all sequenced samples. No trypanosomes were observed by microscopy, and none of the animals showed clinical signs. The results indicate a differential distribution of T. vivax and T. theileri between regions and production systems. Although the study initially focused on T. vivax, the detection of T. theileri highlights the need to consider multiple Trypanosoma species in epidemiological surveys. This study contributes new data on the occurrence of bovine trypanosomes in central Argentina under extensive and semi-intensive management systems. Full article

Background/Objectives: Trypanosomiasis and bovine babesiosis correspond to important diseases that cause great economic losses, but there are still no studies evaluating their occurrence in herds of dairy cattle in Ceará. The aim of this study was to perform molecular diagnosis of the main hemoparasites of dairy cows in the microregion of the central hinterland of Ceará. Methods: For the molecular diagnosis of parasites, genomic material was extracted and polymerase chain reaction directed to the cdCatL-like gene for Trypanossoma vivax and SS rRNA of Babesia bigemina and Babesia. bovis was performed. In addition, the mean corpuscular volume of the samples was evaluated. The data were statistically processed. Results: T. vivax was detected in 0.40% (1/246) of the samples, while B. bigemina and B. bovis were detected in 20.62% (33/160) and 11.87% (19/160) of the samples, respectively. It was found that there was a reduction in mean corpuscular volume in animals that presented with co-infection and those infected by B. bovis only, but not in those hosting B. bigemina alone. The variables “purchase of recent animals” and “tick control” had no association with or influence on B. bovis and/or B. bigemina infection. It was possible to identify epidemiologically important factors that may facilitate the transmission of trypanosoma to healthy animals, such as the recent purchase of animals and use of the same needle and syringe for oxytocin application. Conclusions: The pathogens studied were present in the evaluated population. Daily cow management practices can facilitate the transmission of the diseases they cause. Full article

The potential danger to livestock from African animal trypanosomiasis is well known. However, the trypanosome species circulating in cattle and their genetics are poorly understood. After different alignments according to three regions (ITS1, gGAPDH and rRNA gene) of the trypanosome genome, phylogenetic analyses were used to show the genetic diversity of the different species that were circulating in the cattle in three regions (Bagoue, Poro and Tchologo) of Côte d’Ivoire. These analyses were performed by alignment of ITS1; by alignment of partial 18S, ITS1, 5.8S, ITS2 and partial 28S rRNA genes; and by alignment of gGAPDH gene with sequences of Trypanosomes found in GenBank. Three species were identified (T. vivax, T. theileri and T. congolense) in the cattle in the three northern regions of Côte d’Ivoire. T. vivax and T. theileri were the most abundant species in the present study. Contrary to the other primers used in this study, the ITS1 primers were not able to amplify T. theileri. We observed mixed infections between T. theileri and the other two species identified (T. vivax and T. congolense). As far as primers are concerned, in some cases, rRNA was able to identify the same species of trypanosomes that the ITS1 and gGAPDH primers were able to identify. Two main distinct groups of T. theileri complex were identified. The T. congolense and T. vivax strains were close to African strains, such as those from Kenya, Nigeria and Cameroon, unlike the T. theileri strain. Three trypanosome species (T. vivax, T. theileri and T. congolense) circulate in cattle in the Savannah district of Côte d’Ivoire. The genetic diversity of the trypanosome species encountered in this study cannot be classified as intraspecies according to geographical area and breed of cattle they infect. Full article

The objective of this work was to assess the anemic status and the use of an immunological test and PCR-based methods to determine the infection rates of trypanosomes species. Transhumance aims to provide cattle with greener pastures and greater water resources than in the Djerem region during the dry season. Two criteria were used to assess the health status of the animals, the prevalence of trypanosomiasis and the level of anemia. In addition, we have evaluated the effectiveness, in trypanosomiasis detection, of the Very Diag Kit (CEVA Santé animale), a Rapid diagnosis test (RDT) based on immunological identification of T. congolense s.l. and T. vivax, responsible for AAT. Four trypanosome species (Trypanosoma congolense savannah type (Tcs), T. congolense forest type (Tcf), T. brucei s.l. (Tbr) and T. vivax (Tvx)) were identified in cattle sampled in four villages. The overall infection rate determined by PCR (68.6%) was much higher than those generally reported in cattle from the Adamawa region (35 to 50%). Infections (including mixed infections) by Tc s.l. (Tcs + Tcf) were predominant (45.7%). The infection rates were also determined using the Very Diag Kit allowing us to identify Tc s.l. and Tvx in the field in less than 20 min. This method provided, for the global infection, a higher rate (76.5%) than that determined by PCR (68.6%), although it is supposed to be less sensitive than PCR. Tc s.l. infection rate (37.8%) was similar to that (38.8%) determined by PCR (Tcs + Tcf single infections). In contrast, the prevalence of Tvx single infections measured by RDT (18%) was nearly two-fold higher than that (9.4%) measured by PCR. Thus, further comparative analyses seem to be needed in order to more accurately assess the sensitivity and specificity of the Very Diag test under our conditions of use on blood samples. The mean PCVs in trypanosome-infected as well as in uninfected cattle were below 25%, the threshold below which an animal is considered anemic. Our study shows that cattle return from transhumance in poor health. It raises questions about its real benefit, especially since the herds are themselves likely to become vectors of trypanosomiasis and possibly of other diseases. At least, effective measures have to be undertaken to treat all cattle coming back from transhumance. Full article

Trypanosoma brucei is the causative agent of African trypanosomiasis, a deadly disease that affects humans and cattle. There are very few drugs to treat it, and there is evidence of mounting resistance, raising the need for new drug development. Here, we report the presence of a phosphoinositide phospholipase C (TbPI-PLC-like), containing an X and a PDZ domain, that is similar to the previously characterized TbPI-PLC1. TbPI-PLC-like only possesses the X catalytic domain and does not have the EF-hand, Y, and C2 domains, having instead a PDZ domain. Recombinant TbPI-PLC-like does not hydrolyze phosphatidylinositol 4,5-bisphosphate (PIP₂) and does not modulate TbPI-PLC1 activity in vitro. TbPI-PLC-like shows a plasma membrane and intracellular localization in permeabilized cells and a surface localization in non-permeabilized cells. Surprisingly, knockdown of TbPI-PLC-like expression by RNAi significantly affected proliferation of both procyclic and bloodstream trypomastigotes. This is in contrast with the lack of effect of downregulation of expression of TbPI-PLC1. Full article

African Animal Trypanosomiasis (AAT) is a neglected tropical disease and spreads by the vector tsetse fly, which carries the infectious Trypanosoma sp. in their saliva. Particularly, this parasitic disease affects the health of livestock, thereby imposing economic constraints on farmers, costing billions of dollars every year, especially in sub-Saharan African countries. Mainly considering the AAT disease as a multistage progression process, we previously performed upstream analysis to identify transcription factors (TFs), their co-operations, over-represented pathways and master regulators. However, downstream analysis, including effectors, corresponding gene expression profiles and their association with the regulatory SNPs (rSNPs), has not yet been established. Therefore, in this study, we aim to investigate the complex interplay of rSNPs, corresponding gene expression and downstream effectors with regard to the AAT disease progression based on two cattle breeds: trypanosusceptible Boran and trypanotolerant N’Dama. Our findings provide mechanistic insights into the effectors involved in the regulation of several signal transduction pathways, thereby differentiating the molecular mechanism with regard to the immune responses of the cattle breeds. The effectors and their associated genes (especially MAPKAPK5, CSK, DOK2, RAC1 and DNMT1) could be promising drug candidates as they orchestrate various downstream regulatory cascades in both cattle breeds. Full article

The parasite Trypanosoma brucei (T. brucei) is responsible for human African trypanosomiasis (HAT) and the cattle disease “Nagana” which to this day cause severe medical and socio-economic issues for the affected areas in Africa. So far, most of the available treatment options are accompanied by harmful side effects and are constantly challenged by newly emerging drug resistances. Since trypanosomatids are auxotrophic for folate, their pteridine metabolism provides a promising target for an innovative chemotherapeutic treatment. They are equipped with a unique corresponding enzyme system consisting of the bifunctional dihydrofolate reductase-thymidylate synthase (TbDHFR-TS) and the pteridine reductase 1 (TbPTR1). Previously, gene knockout experiments with PTR1 null mutants have underlined the importance of these enzymes for parasite survival. In a search for new chemical entities with a dual inhibitory activity against the TbPTR1 and TbDHFR, a multi-step in silico procedure was employed to pre-select promising candidates against the targeted enzymes from a natural product database. Among others, the sesquiterpene lactones (STLs) cynaropicrin and cnicin were identified as in silico hits. Consequently, an in-house database of 118 STLs was submitted to an in silico screening yielding 29 further virtual hits. Ten STLs were subsequently tested against the target enzymes in vitro in a spectrophotometric inhibition assay. Five compounds displayed an inhibition over 50% against TbPTR1 as well as three compounds against TbDHFR. Cynaropicrin turned out to be the most interesting hit since it inhibited both TbPTR1 and TbDHFR, reaching IC₅₀ values of 12.4 µM and 7.1 µM, respectively. Full article

Toxoplasma gondii is an apicomplexan parasite that infects and proliferates within many different types of host cells and infects virtually all warm-blooded animals and humans. Trypanosoma brucei is an extracellular kinetoplastid that causes human African trypanosomiasis and Nagana disease in cattle, primarily in rural sub-Saharan Africa. Current treatments against both parasites have limitations, e.g., suboptimal efficacy and adverse side effects. Here, we investigate the potential cellular and molecular targets of a trithiolato-bridged arene ruthenium complex conjugated to 9-(2-hydroxyethyl)-adenine (1), which inhibits both parasites with IC₅₀s below 10⁻⁷ M. Proteins that bind to 1 were identified using differential affinity chromatography (DAC) followed by shotgun-mass spectrometry. A trithiolato-bridged ruthenium complex decorated with hypoxanthine (2) and 2-hydroxyethyl-adenine (3) were included as controls. Transmission electron microscopy (TEM) revealed distinct ultrastructural modifications in the mitochondrion induced by (1) but not by (2) and (3) in both species. DAC revealed 128 proteins in T. gondii and 46 proteins in T. brucei specifically binding to 1 but not 2 or 3. In T. gondii, the most abundant was a protein with unknown function annotated as YOU2. This protein is a homolog to the human mitochondrial inner membrane translocase subunit Tim10. In T. brucei, the most abundant proteins binding specifically to 1 were mitochondrial ATP-synthase subunits. Exposure of T. brucei bloodstream forms to 1 resulted in rapid breakdown of the ATP-synthase complex. Moreover, both datasets contained proteins involved in key steps of metabolism and nucleic acid binding proteins. Full article

African animal trypanosomiasis (AAT) control programs rely on active case detection through the screening of animals reared in disease endemic areas. This study compared the application of the polymerase chain reaction (PCR) and microscopy in the detection of trypanosomes in cattle blood in Mambwe, a rural district in eastern Zambia. Blood samples were collected from 227 cattle and tested for infection with trypanosomes using microscopy and Ribosomal RNA Internal Transcribed Spacers (ITS)-PCR. Microscopy on the buffy coat detected 17 cases, whilst thin and thick smears detected 26 cases and 28 cases, respectively. In total, microscopy detected 40 cases. ITS-PCR-filter paper (FP) on blood spots stored on FP detected 47 cases, and ITS-PCR-FTA on blood spots stored on Whatman FTA Classic cards detected 83 cases. Using microscopy as the gold standard, ITS-PCR-FTA had a better specificity (SP) and sensitivity (SE) (SP = 72.2%; SE = 77.5%; kappa = 0.35) than ITS-PCR-FP (SP = 88%; SE = 60%; kappa = 0.45). The prevalence of Trypanosoma brucei s.l. was higher on ITS-PCR-FTA (19/227) than on ITS-PCR-FP (0/227). Our results illustrate the complexities around trypanosomiasis surveillance in rural Africa and provide evidence of the impact that field conditions and staff training can have on diagnostic results, which in turn impact the success of tsetse and trypanosomiasis control programs in the region. Full article

African Animal Trypanosomiasis (AAT) is transmitted by the tsetse fly which carries pathogenic trypanosomes in its saliva, thus causing debilitating infection to livestock health. As the disease advances, a multistage progression process is observed based on the progressive clinical signs displayed in the host’s body. Investigation of genes expressed with regular monotonic patterns (known as Monotonically Expressed Genes (MEGs)) and of their master regulators can provide important clue for the understanding of the molecular mechanisms underlying the AAT disease. For this purpose, we analysed MEGs for three tissues (liver, spleen and lymph node) of two cattle breeds, namely trypanosusceptible Boran and trypanotolerant N’Dama. Our analysis revealed cattle breed-specific master regulators which are highly related to distinguish the genetic programs in both cattle breeds. Especially the master regulators MYC and DBP found in this study, seem to influence the immune responses strongly, thereby susceptibility and trypanotolerance of Boran and N’Dama respectively. Furthermore, our pathway analysis also bolsters the crucial roles of these master regulators. Taken together, our findings provide novel insights into breed-specific master regulators which orchestrate the regulatory cascades influencing the level of trypanotolerance in cattle breeds and thus could be promising drug targets for future therapeutic interventions. Full article

Tsetse transmitted trypanosomiasis is a fatal disease commonly known as Nagana in cattle and sleeping sickness in humans. The disease threatens food security and has severe economic impact in Africa including most parts of Zambia. The level of effectiveness of commonly used African trypanosomiasis control methods has been reported in several studies. However, there have been no review studies on African trypanosomiasis control and management conducted in the context of One Health. This paper therefore seeks to fill this knowledge gap. A review of studies that have been conducted on African trypanosomiasis in Zambia between 2009 and 2019, with a focus on the control and management of trypanosomiasis was conducted. A total of 2238 articles were screened, with application of the search engines PubMed, PubMed Central and One Search. Out of these articles, 18 matched the required criteria and constituted the basis for the paper. An in-depth analysis of the 18 articles was conducted to identify knowledge gaps and evidence for best practices. Findings from this review provide stakeholders and health workers with a basis for prioritisation of African trypanosomiasis as an important neglected disease in Zambia and for formulation of One Health strategies for better control and/or management of the disease. Full article

African Animal Trypanosomiasis (AAT) is a disease caused by pathogenic trypanosomes which affects millions of livestock every year causing huge economic losses in agricultural production especially in sub-Saharan Africa. The disease is spread by the tsetse fly which carries the parasite in its saliva. During the disease progression, the cattle are prominently subjected to anaemia, weight loss, intermittent fever, chills, neuronal degeneration, congestive heart failure, and finally death. According to their different genetic programs governing the level of tolerance to AAT, cattle breeds are classified as either resistant or susceptible. In this study, we focus on the cattle breeds N’Dama and Boran which are known to be resistant and susceptible to trypanosomiasis, respectively. Despite the rich literature on both breeds, the gene regulatory mechanisms of the underlying biological processes for their resistance and susceptibility have not been extensively studied. To address the limited knowledge about the tissue-specific transcription factor (TF) cooperations associated with trypanosomiasis, we investigated gene expression data from these cattle breeds computationally. Consequently, we identified significant cooperative TF pairs (especially $DBP-PPARA$ and $DBP-THAP1$ in N’Dama and $DBP-PAX8$ in Boran liver tissue) which could help understand the underlying AAT tolerance/susceptibility mechanism in both cattle breeds. Full article

African animal trypanosomiasis is caused by vector-transmitted parasites of the genus Trypanosoma. T. congolense and T. brucei brucei are predominant in Africa; T. evansi and T. vivax in America and Asia. They have in common an extracellular lifestyle and livestock tropism, which provokes huge economic losses in regions where vectors are endemic. There are licensed drugs to treat the infections, but adherence to treatment is poor and appearance of resistances common. Therefore, the availability of a prophylactic vaccine would represent a major breakthrough towards the management and control of the disease. Selection of the most appropriate antigens for its development is a bottleneck step, especially considering the limited resources allocated. Herein we propose a vaccine strategy based on multiple epitopes from multiple antigens to counteract the parasites´ biological complexity. Epitopes were identified by computer-assisted genome-wide screenings, considering sequence conservation criteria, antigens annotation and sub-cellular localization, high binding affinity to antigen presenting molecules, and lack of cross-reactivity to proteins in cattle and other breeding species. We ultimately provide 31 B-cell, 8 CD4 T-cell, and 15 CD8 T-cell epitope sequences from 30 distinct antigens for the prospective design of a genetic ensemble vaccine against the four trypanosome species responsible for African animal trypanosomiasis. Full article

We carried out a baseline survey of cattle in Kaberamaido district, in the context of controlling the domestic animal reservoir of Trypanosoma brucei rhodesiense human African trypanosomiasis (rHAT) towards elimination. Cattle blood was subjected to capillary tube centrifugation followed by measurement of the packed cell volume (PCV) and examination of the buffy coat area for motile trypanosomes. Trypanosomes were detected in 561 (21.4%) out of 2621 cattle screened by microscopy. These 561 in addition to 724 apparently trypanosome negative samples with low PCVs (≤25%) were transported to the laboratory and tested by PCR targeting the trypanosomal Internal Transcribed Spacer (ITS-1) as well as suspect Tick-Borne Diseases (TBDs) including Anaplasmamosis, Babesiosis, and Theileriosis. PCR for Anaplasma sp yielded the highest number of positive animals (45.2%), followed by Trypanosoma sp (44%), Theileria sp (42.4%) and Babesia (26.3%); multiple infections were a common occurrence. Interestingly, 373 (29%) of these cattle with low PCVs were negative by PCR, pointing to other possible causes of aneamia, such as helminthiasis. Among the trypanosome infections classified as T. brucei by ITS-PCR, 5.5% were positive by SRA PCR, and were, therefore, confirmed as T. b. rhodesiense. Efforts against HAT should therefore consider packages that address a range of conditions. This may enhance acceptability and participation of livestock keepers in programs to eliminate this important but neglected tropical disease. In addition, we demonstrated that cattle remain an eminent reservoir for T. b. rhodesiense in eastern Uganda, which must be addressed to sustain HAT elimination. Full article

Increasing the production of meat and milk within sub-Saharan Africa should provide significant food security benefits. However, greenhouse gas (GHG) emissions represent a challenge, as cattle production in the region typically has high emissions intensity (EI), i.e., high rates of GHG emissions per unit of output. The high EI is caused by the relatively low production efficiencies in the region, which are in turn partly due to endemic cattle diseases. In theory, improved disease control should increase the efficiency and decrease the emissions intensity of livestock production; however quantitative analysis of the potential GHG mitigation effects of improved disease control in Africa is lacking. This paper seeks to respond to this by using a hybrid modelling approach to quantify the production and emissions effects of removing trypanosomiasis from East African cattle production systems. The emissions are quantified for each cattle production system using an excel version of GLEAM, the Food and Agriculture Organization’s Global Livestock Environmental Assessment Model. The results indicate that removing trypanosomiasis leads to a reduction in the emissions intensity per unit of protein produced of between 0% and 8%, driven mainly by the increases in milk yields and cow fertility rates. Despite the limitations, it is argued that the approach provides considerable scope for modelling the GHG impacts of disease interventions. Full article