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Background: Trypanosomiasis continues to pose a global threat to human health, with human infection mainly caused by Trypanosoma brucei and Trypanosoma cruzi., Methods: We present a 30-year-old pregnant woman with persistent high fever from Shandong Province, China. High-throughput sequencing revealed the presence of Trypanosoma dionisii in blood. We conducted an analysis of the patient's clinical, epidemiological, and virological data., Results: The patients exhibited fever, shortness of breath, chest tightness, accompanied by change in liver function and inflammatory response. She made a full recovery without any long-term effects. T. dionisii was detected in blood collected 23 days after onset of illness. The 18S rRNA gene sequence showed close similarity to T. dionisii found in bats from Japan, while the gGAPDH gene was closely related to T. dionisii from bats in Mengyin County, Shandong Province. Phylogenetic analysis demonstrated the current T. dionisii belongs to clade B within its species group. Positive anti-Trypanosoma IgG antibody was detected from the patient on Day 23, 66 and 122 after disease onset, as well as the cord blood and serum from the newborn. Retrospective screening of wild small mammals captured from Shandong Province revealed a prevalence rate of 0.54% (7/1304) for T. dionisii; specifically among 0.81% (5/620) of Apodemus agrarius, and 0.46% (2/438) of Mus musculus., Conclusions: The confirmation of human infection with T. dionisii underscores its potential as a zoonotic pathogen, while the widespread presence of this parasite in rodent and bat species emphasizes the emerging threat it poses to human health., Competing Interests: Declaration of Competing Interest The authors declare no conflicts of interest., (Copyright © 2024. Published by Elsevier Ltd.)

Background: Surra is a parasitic disease caused by Trypanosoma evansi that threatens the health and productivity of camels. Despite its significant impact on camels in Ethiopia, surra has not received as much attention as diseases in cattle and other domestic animals. The objective of the study was to estimate the prevalence of surra, identify the potential risk factors and assess the traditional knowledge, attitude and practices of camel herders towards the disease., Methods: The study used a parasitological and participatory epidemiological (PE) approach. Between February and July 2022, a total of 335 blood samples were collected from camels across three districts and tested using the buffy coat technique. The PE investigation involved six key informant groups consisting of 8 to 12 key persons, and used a semi-structured interview and various PE tools and principles., Result: The study found that the prevalence of surra among examined camels was 3.9% (95% CI: 2.1-6.5). The prevalence was significantly higher in camels with a poor body condition score (BCS) (OR = 9.3; 95% CI: 1.8-47.5; p = 0.008) compared with camels with a good BCS. However, district, age, sex, and ethnicity had no effect on the prevalence of surra (p > 0.05). The study also found that the packed cell volume (PCV) was significantly lower (p < 0.0001) in parasitaemic animals (18.92 ± 2.63) than in aparasitaemic animals (25.13 ± 4.56). Camels with poor BCS (22.7 ± 3.5) had a significantly (p < 0.001) lower mean PCV than camels with good BCS (26.2 ± 5.0). The PE investigation showed that all the camel herders were well aware of surra, known locally as Dhukana. The clinical symptoms, the season of high incidence, routes of transmission, impact on production, and control methods were accurately described. Moreover, this study emphasized that surra is the primary disease affecting camel health and productivity., Conclusion: The study identified a moderate prevalence of surra in the research area. To reduce surra incidence and associated losses, enhancing veterinary services and providing support for proper camel husbandry practices in the region is recommended. Additionally, future studies should consider using more sensitive and specific techniques like serological and molecular assays, as this study relied on microscopy only., (© 2024. The Author(s).)

The present research aimed to document the incidence, clinical signs, haematological, and serum biochemical alterations, as well as electrocardiography and echocardiography findings in 62 buffaloes (selected from a total of 240) infected with Trypanosoma evansi. The study spanned one year, from January 2022 to December 2022. Morphological identification of Trypanosoma evansi was done by the presence of a centrally positioned nucleus with a small sub-terminal kinetoplast at the posterior position through microscopic examination of Giemsa stained peripheral blood smears. The incidence of trypanosomosis were determined to be 26% (62/240) using stained blood smear examination and 41% (98/240) through polymerase chain reaction assay. Clinical signs exhibited by buffaloes with trypanosomosis included the lack of rumination (94%; 58/62), anorexia (90%; 56/62), emaciation (87%; 54/62), loss of milk yield (84%; 52/62), ocular discharges (82%; 51/62), depressed demeanour (81%; 50/62), sunken eye balls (61%; 38/62), fever (60%; 37/62), scleral congestion (56%; 35/62) and intermittent fever (42%; 26/62). Cardiovascular clinical findings in affected buffaloes included tachycardia (44%; 27/62), cardiac arrhythmia (24%; 15/62), cardiac murmurs (19%; 12/62) and muffled heart sounds (18%; 11/62). In the present study, buffaloes with trypanosomosis exhibited significant reduction in haemoglobin (p = 0.008), packed cell volume (p = 0.004), total erythrocyte count (p = 0.003), mean corpuscular volume (p = 0.042), total leucocyte count (p = 0.048) and absolute neutrophil count (p = 0.012); a significant increase in absolute eosinophil count (p = 0.011) and absolute monocyte count (p = 0.008) compared to the apparently healthy buffaloes. Additionally significant decrease in albumin (p = 0.001), A/G ratio (p = 0.007), calcium (p = 0.008), glucose (p = 0.007), phosphorous (p = 0.048), sodium (p = 0.008), potassium (p = 0.041) and chloride (p = 0.046) were observed in buffaloes with trypanosomosis compared to healthy ones. Buffaloes with trypanosomosis also showed significant increase in globulin (p = 0.004), aspartate aminotransferase (p = 0.008), bilirubin (p = 0.034), blood urea nitrogen (p = 0.071), creatinine (p = 0.029), cholesterol (p = 0.046), lactate dehydrogenase (p = 0.009), gamma-glutamyl transferase (p = 0.004) and creatine kinase-myoglobin binding levels (p = 0.005). Electrocardiography explorations in buffaloes with trypanosomosis revealed sinus tachycardia, low voltage QRS complex, ST segment elevation, wide QRS complex, sinus arrhythmia, sinus bradycardia, wandering pace maker, first degree atrio ventricular block, biphasic T wave and tall T wave. Echocardiography examination unveiled cardiac chamber dilatation, ventricular wall thickening and indications of pericarditis/cardiac tamponade. Necropsy was carried on the dead buffaloes during the study period disclosed severely congested blood vessels on epicardial surface, endocardial haemorrhages, and presence of pericardial fluid. Histopathological examination of the heart revealed hyaline degeneration, haemorrhages in the cardiac muscles and varying degrees of degenerative changes. Additionally, the pericardium displayed increased thickness due to presence of more elastic fibres, fibroblast cells in the myocardium, discontinuity of muscle layers, vascular congestion, perivascular mono nuclear cell infiltration and augmented thickness of the endocardium with fibroblast cell proliferation. The study's conclusion highlights cardiac alterations as secondary complications in buffaloes infected with Trypanosoma evansi. Further investigations are recommended to elucidate therapeutic modifications and refine the treatment paradigm., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Trypanosomosis due to Trypanosoma evansi (surra) is one of the most important diseases with a significant impact on camel health and production. Trypanosoma-induced immunosuppression mechanisms, which are key factors of disease pathogenesis, have been characterized in several animal species. The present study investigated, therefore, the impact of trypanosomosis on the immunophenotype of blood leukocytes in camels. For this, the relative and absolute values of blood leukocyte populations, their expression pattern of cell surface molecules, and the numbers of the main lymphocyte subsets were compared between healthy camels and camels with clinical symptoms of chronic surra and serological evidence of exposure to Trypanosoma infection. Leukocytes were separated from the blood of healthy and diseased camels, labeled with fluorochrome-conjugated antibodies, and analyzed by flow cytometry. Compared to healthy camels, the leukogram of diseased camels was characterized by a slightly increased leukocyte count with moderate neutrophilia and monocytosis indicating a chronic inflammatory pattern that may reflect tissue injury due to the long-lasting inflammation. In addition, the analysis of lymphocyte subsets revealed a lower number and percentage of B cells in diseased than healthy camels. In vitro incubation of camel mononuclear cells with fluorochrome-labeled T. evansi revealed a higher capacity of camel B cells than T cells to bind the parasite in vitro. Furthermore, cell viability analysis of camel PBMC incubated in vitro with T. evansi whole parasites but not the purified antigens resulted in Trypanosoma-induced apoptosis and necrosis of camel B cells. Here we demonstrate an association between trypanosomosis in camels and reduced numbers of blood B cells. In vitro analysis supports a high potential of T. evansi to bind to camel B cells and induce their elimination by apoptosis and necrosis., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Bats are hosts for diverse Trypanosoma species, including trypanosomes of the Trypanosoma cruzi clade. This clade is believed to have originated in Africa and diversified in many lineages worldwide. In several geographical areas, including Cameroon, no data about trypanosomes of bats has been collected yet. In this study, we investigated the diversity and phylogenetic relationships of trypanosomes of different bat species in the central region of Cameroon. Trypanosome infections were detected in six bat species of four bat families, namely Hipposideridae, Pteropodidae, Rhinolophidae, and Vespertilionidae, with an overall prevalence of 29% and the highest infection rate in hipposiderid bat species. All trypanosomes were identified as belonging to the Trypanosoma livingstonei species group with one clade that might represent an additional subspecies of T. livingstonei. Understanding the prevalence, distribution, and host range of parasites of this group contributes to our overall knowledge of the diversity and host specificity of trypanosome species that phylogenetically group at the base of the T. cruzi clade., (© 2024. The Author(s).)

Animal trypanosomosis, caused by Trypanozoon trypanosomes (Trypanosoma evansi and T. equiperdum), and Trypanosoma vivax, is endemic to South American countries and has a negative impact on the livestock industry. However, the risk factors for trypanosomosis in Paraguay remain unknown. This study aimed to determine the risk factors for equine trypanosomosis in Paraguay based on a PCR-based molecular survey and individual horse sampling data. In this study, 739 blood samples were collected from horses in 16 departments of Paraguay between August 2019 and November 2020. To elucidate the risk factors for trypanosome infection, the relationship between trypanosome infection status detected by PCR and the location, sex, age, breed of horses, and season of sample collection was analyzed. There were no significant differences in trypanosome prevalence in horses between the eastern and western regions, ages, or breeds of horses in Paraguay. Sex and season were identified as risk factors for trypanosome infection in horses in Paraguay in the current study. These results suggest that the rainy-summer season, when vectors increase in number and their blood-sucking activity, could be the most important risk factor for trypanosome infection in Paraguay horses. Preventive measures and treatments should be developed to address these factors., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Trypanosomes are protozoan parasites of class Kinetoplastida. Trypanosoma vivax is one of the organisms that can cause Nagana and Trypanosoma evansi can cause Surra. In Africa, Trypanosoma vivax is mainly transmitted by Glossina spp. (tsetse fly) but it can be transmitted mechanically by other blood-feeding dipters. Trypanosoma evansi is transmitted mechanically and non-dependent to tsetse fly. In this research, T. vivax and T. evansi among camels (Camelus dromedarius) in Yazd, Iran were identified by microscopy and molecular examinations but the sensitivity of microscopy was lower than molecular examinations. Trypanosoma vivax and T. evansi were observed in 4 out of 134 blood film samples (2.98%). The prevalence of Trypanosoma spp. among 134 male camels (C. dromedarius) based on molecular examinations was 30.6% (22.76-38.44% with 95% confidence interval), 25 out of 134 (18.65%) had co-infection of T. evansi and T. vivax, and 16 out of 134 (11.94%) had an infection of T. vivax alone. We provided the first confirmation of infection with T. vivax among camels in Iran, and also in Asia, which has important implications on our knowledge of the occurrence and possible spread of this pathogen at the global level. Investigations in other species such as cattle and sheep are strongly recommended., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Tabanids (syn. horse flies) are biting-flies of medical and veterinary significance because of their ability to transmit a range of pathogens including trypanosomes - some species of which carry a combined health and biosecurity risk. Invertebrate vectors responsible for transmitting species of Trypanosoma between Australian wildlife remains unknown, thus establishing the role of potential vector candidates such as tabanids is of utmost importance. The current study aimed to investigate the presence of indigenous trypanosomes in tabanids from an endemic area of south-west Australia. A total of 148 tabanids were collected, with morphological analysis revealing two subgenera: Scaptia (Pseudoscione) and S. (Scaptia) among collected flies. A parasitological survey using an HRM-qPCR and sequencing approach revealed a high (105/148; 71%) prevalence of trypanosomatid DNA within collected tabanids. Individual tissues - proboscis (labrum, labium and mandibles, hypopharynx), salivary glands, proventriculus, midgut, and hindgut and rectum - were also tested from a subset of 20 tabanids (n = 140 tissues), confirming the presence of Trypanosoma noyesi in 31% of screened tissues, accompanied by T. copemani (3%) and T. vegrandis/T.gilletti (5%). An unconfirmed trypanosomatid sp. was also detected (9%) within tissues. The difference between tissues infected with T. noyesi compared with tissues infected with other trypanosome species was statistically significant (p < 0.05), revealing T. noyesi as the more frequent species detected in the tabanids examined. Fluorescence in situ hybridisation (FISH) and scanning electron microscopy (SEM) confirmed intact parasites within salivary glands and the proboscis respectively, suggesting that both biological and mechanical modes of transmission could occur. This study reveals the presence of Australian Trypanosoma across tabanid tissues and confirms intact parasites within tabanid salivary glands and the proboscis for the first time. Further investigations are required to determine whether tabanids have the vectorial competence to transmit Australian trypanosomes between wildlife., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

In South America, Colombia is the third-largest livestock producer with approximately 28.8 million cattle, of which Colombian Creole cattle represent around 1% of the livestock population. Animal Trypanosomiasis (AT) is one of the most critical problems in the livestock industry, reducing its production by about 30 %. Considering the paucity of information to understand the epidemiological features of AT in Colombian Creole cattle, the present study reports the molecular prevalence and clinical traits associated with the infection of Trypanosoma spp. in three Colombian Creole breeds. From 2019 to 2020, cross-sectional surveillance in farms of central and west of Colombia was designed to evaluate the mentioned characteristics in Casanareño, Chino Santandereano, and Sanmartinero Creole breeds. Molecular analysis showed an AT prevalence of 60.2 % (95 % CI = 54.2 % - 66.2 %). The Chino Santandereano population presented the highest value (Trypanosoma spp., 75.2 %, T. theileri 59.6 % and T. evansi 15.6 %), followed by Casanareño (Trypanosoma spp., 65.3 %, T. theileri 38.6 %, T. evansi 24.0 %, and T. vivax 5.3 %) and Sanmartinero (Trypanosoma spp., 33.3 %, T. theileri 24.0 % and T. evansi 9.3 %). Features such as breeds, age, and feeding system were significantly associated with AT prevalence (P <  0.05). Additionally, a low level of serum total proteins was observed during T. evansi infection in Sanmartinero (P <  0.05). To our knowledge, this is the first cross-sectional survey that evaluates using molecular methods the infection of Trypanosoma spp. in Colombian Creole breeds, showing significant variations in the prevalence and clinical signs associated with the infection. These results suggest different degrees of trypanotolerance in these breeds, as well as a possible effect of environmental variables on the prevalence and clinical characteristics associated with the infection. The epidemiological and economic implications of these findings are discussed here., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Trypanosoma brucei evansi and T. brucei equiperdum are animal infective trypanosomes conventionally classified by their clinical disease presentation, mode of transmission, host range, kinetoplast DNA (kDNA) composition and geographical distribution. Unlike other members of the subgenus Trypanozoon , they are non-tsetse transmitted and predominantly morphologically uniform (monomorphic) in their mammalian host. Their classification as independent species or subspecies has been long debated and genomic studies have found that isolates within T. brucei evansi and T. brucei equiperdum have polyphyletic origins. Since current taxonomy does not fully acknowledge these polyphyletic relationships, we re-analysed publicly available genomic data to carefully define each clade of monomorphic trypanosome. This allowed us to identify, and account for, lineage-specific variation. We included a recently published isolate, IVM-t1, which was originally isolated from the genital mucosa of a horse with dourine and typed as T. equiperdum . Our analyses corroborate previous studies in identifying at least four distinct monomorphic T. brucei clades. We also found clear lineage-specific variation in the selection efficacy and heterozygosity of the monomorphic lineages, supporting their distinct evolutionary histories. The inferred evolutionary position of IVM-t1 suggests its reassignment to the T. brucei evansi type B clade, challenging the relationship between the Trypanozoon species, the infected host, mode of transmission and the associated pathological phenotype. The analysis of IVM-t1 also provides, to our knowledge, the first evidence of the expansion of T. brucei evansi type B, or a fifth monomorphic lineage represented by IVM-t1, outside of Africa, with important possible implications for disease diagnosis.

This is the first case report for the positive Trypanosoma evansi incident in Kelantan, Malaysia confirmed through protozoa detection in a Siam B mare. The horse was presented with complaints of lethargy and inappetence and it collapsed on the day of visit. Normal saline and dextrose solution were administered intravenously, while multivitamins and nerve supplements were given intramuscularly to stabilise the horse before further treatment. Haematological findings showed normocytic hypochromic anaemia and are suggestive of regenerative anaemia. Thin blood smear and examination revealed the presence of Trypanosoma sp., and it was confirmed as T. evansi through molecular identification. The horse was found dead 2 days after and post-mortem was conducted. Histopathology revealed that the horse had developed a neurological form of the disease, besides the detection of the protozoa in heart, spleen and kidney tissue. This first positive Surra case in Kelantan, Malaysia, that is bordering Thailand confirms the increasing concern of transboundary infections. In conclusion, Surra is a potential emerging disease and should be considered as differential diagnosis in horses with pale mucous membrane. This condition is particularly imperative in horses found in these regions as Surra is endemic., (© 2020 The Authors Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Parasitic protozoan trypanosomes of the genus Trypanosoma cause infections in both man and livestock in Africa. Understanding the current spatial distribution of trypanosomes, herd-level factors associated with Trypanosoma brucei infection as well as local knowledge of African trypanosomosis is key to its prevention and control. A cross-sectional study was performed that sampled 53 livestock farmers and 444 cattle throughout Malawi. Cattle were screened for trypanosomes using serology and molecular techniques. Questionnaires were administered to livestock herders and incidence of hospital diagnosed human trypanosome infections was estimated from reports submitted to the Department of Health Unit. The apparent prevalence of trypanosome species based on molecular detection was low for Trypanosoma brucei (2%; 95 % CI: 1-4 %) and Trypanosoma congolense (3%; 95 % CI: 2-5 %) but higher for Trypanosoma theileri (26 %; 95 % CI: 22-30 %). The central region of the country was identified as being at a higher risk of T.brucei infection. One of the sampled cattle was confirmed as being infected with Trypanosoma brucei rhodesiense. Human trypanosome cases were more frequently reported in the northern region with an estimated incidence of 5.9 cases per 100,000 people in Rumphi District. The control of zoonotic diseases that impact poor livestock herders requires a One Health approach due to the close contact between humans and their animals and the reliance on animal production for a sustainable livelihood., (Copyright © 2020. Published by Elsevier B.V.)

A 2.5-year-old male German Shepherd was presented to a private veterinary clinic in Hanoi, Vietnam showing anorexia, weakness, lethargy, reluctant to go for walks with a recent history of intermittent fever. Clinical examination of the dog showed pale mucous membrane, impaired eyesight, edema of the back legs. Complete blood count revealed severe anemia; red blood cell 3.8 × 10 12 /l, hemoglobin 8.7 g/dl, hematocrit 26.4%, associated with thrombocytopenia 145 × 10 9 /l. Biochemical analysis showed a moderate increase of alanine transaminase (150.7 UI/l) and alkaline phosphatase activities (266 UI/I) with mild hypoglycemia (71.46 mg/dl). Trypanosoma evansi was observed in Giemsa-stained blood smears under microscopic observation which was confirmed by PCR. This is the first report of canine trypanosomiasis caused by T. evansi in Vietnam., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Surra, caused by Trypanosoma evansi, is a re-emerging animal trypanosomosis, which is of special concern for camel-rearing regions of Africa and Asia. Surra decreases milk yield, lessens animal body condition score and reduces market value of exported animals resulting in substantial economic losses. A cross-sectional seroprevalence study of dromedary camels was conducted in Algeria, and major risk factors associated with infection were identified by collecting data on animal characteristics and herd management practices. The seroprevalence of T. evansi infection was determined in sera of 865 camels from 82 herds located in eastern Algeria using an antibody test (card agglutination test for Trypanosomiasis - CATT/T. evansi). Individual and herd seroprevalence were 49.5% and 73.2%, respectively, indicating substantial exposure of camels to T. evansi in the four districts studied. Five significant risk factors for T. evansi hemoparasite infection were identified: geographical area, herd size, husbandry system, accessibility to natural water sources and type of watering. There was no association between breed, sex or age with T. evansi infection. Results of this study provide baseline information that will be useful for launching control programmes in the region and potentially elsewhere.

Trypanosoma evansi (T. evansi) is a flagellated parasite with worldwide distribution, mainly affecting camels, horses, dogs, buffaloes and wild animals. Trypanosomosis caused by T. evansi, known as surra, is a vector borne disease that affects the health and productivity of camels. The aim of our study was to assess the prevalence of trypanosomosis due to T. evansi in camels by the immune trypanolosis test and to identify associated risk factors. Our cross-sectional study was performed on 161 camels from Ghardaïa district, southern Algeria. A structured questionnaire was used to collect data on individual characteristics (age, gender and breed) husbandry management (herd size and activity of animals) and health conditions (history of abortion and clinical symptoms). The immune trypanolysis test revealed an overall seroprevalence of 9.3% (CI 95%, 5.9-14.9). Possible factors associated with T. evansi infection were analysed by univariate and multivariate logistic regression. The results showed that risk factors for camels were history of symptoms (P = 0.002, OR = 21.91, CI95% = 3.48-169.80), racing activities (P = 0.003, OR = 0.01, CI95% = 0.001-0.18) and small herd size (P = 0.013, OR = 8.22, CI95% = 1.64-49.75). In conclusion, this study showed that T. evansi is endemic in camels of Ghardaïa district. To reduce dissemination of the disease to non-endemic areas, it is recommended to minimise risk factors associated with the infection., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Trypanosomes are blood-borne parasites that can infect a variety of different vertebrates, including animals and humans. This study aims to broaden scientific knowledge about the presence and biodiversity of trypanosomes in Australian bats. Molecular and morphological analysis was performed on 86 blood samples collected from seven different species of microbats in Western Australia. Phylogenetic analysis on 18S rDNA and glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) sequences identified Trypanosoma dionisii in five different Australian native species of microbats; Chalinolobus gouldii, Chalinolobus morio, Nyctophilus geoffroyi, Nyctophilus major and Scotorepens balstoni. In addition, two novels, genetically distinct T. dionisii genotypes were detected and named T. dionisii genotype Aus 1 and T. dionisii genotype Aus 2. Genotype Aus 2 was the most prevalent and infected 20.9% (18/86) of bats in the present study, while genotype Aus 1 was less prevalent and was identified in 5.8% (5/86) of Australian bats. Morphological analysis was conducted on trypomastigotes identified in blood films, with morphological parameters consistent with trypanosome species in the subgenus Schizotrypanum. This is the first report of T. dionisii in Australia and in Australian native bats, which further contributes to the global distribution of this cosmopolitan bat trypanosome.

Several species of trypanosomes can infect bats (Chiroptera), but current information about bat trypanosomes in Colombia is scarce. The objectives of this study were to estimate the infection rate and to characterize the trypanosome species infecting bats from three rural regions near the municipality of Cumaribo in Vichada, Colombia. Blood samples were collected from 39 bats. DNA was extracted from the blood samples and analyzed using nuclear genetic markers (SSU rDNA, ITS rDNA, and cathepsin genes) to discriminate among trypanosome species. Trypanosomes were detected in 66.7 % (26/39) of blood samples using PCR; 61.5 % (24/39) of infections were identified as Trypanosoma theileri and 5.1 % (2/39) as T. wauwau-like parasites. The phylogeographic analysis revealed that our T. theileri sequences were associated with the TthIIB genotype from cattle in Brazil and Venezuela. The T. wauwau-like parasites represent a new genotype of the species and were found in Molossus molossus and Platyrrhinus helleri bats. These data represent the first evidence of this trypanosome in both Colombia, and in these species of bats. Bat infections with T. theileri suggest an important role of these hosts in maintaining this genotype, probably acquired by ingesting insect vectors. The T. wauwau-like genotype in new mammalian host species supports the 'bat seeding' hypothesis of the T. cruzi clade. The epidemiological and evolutionary implications of these findings are discussed., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The Indonesian island of Sulawesi is a globally significant biodiversity hotspot with substantial undescribed biota, particularly blood-borne parasites of endemic wildlife. Documenting the blood parasites of Sulawesi's murine rodents is the first fundamental step towards the discovery of pathogens likely to be of concern for the health and conservation of Sulawesi's endemic murines. We screened liver samples from 441 specimens belonging to 20 different species of murine rodents from 2 mountain ranges on Sulawesi, using polymerase chin reaction (PCR) primers targeting the conserved 18S rDNA region across the protozoan class Kinetoplastea. We detected infections in 156 specimens (10 host species) with a mean prevalence of 35.4% (95% confidence interval [CI] = 30.9-39.8%). Sequences from these samples identified 4 infections to the genus Parabodo, 1 to Blechomonas, and the remaining 151 to the genus Trypanosoma. Within Trypanosoma, we recovered 17 haplotypes nested within the Trypanosoma theileri clade infecting 117 specimens (8 host species) and 4 haplotypes nested within the Trypanosoma lewisi clade infecting 34 specimens (6 host species). Haplotypes within the T. theileri clade were related to regional Indo-Australian endemic trypanosomes, displayed geographic structuring but with evidence of long-term connectivity between mountains, and had substantial phylogenetic diversity. These results suggest T. theileri clade parasites are native to Sulawesi. Conversely, T. lewisi clade haplotypes were recovered from both endemic and introduced rodents, demonstrated complete geographic separation between clades, and had low genetic diversity. These results suggest that the T. lewisi clade parasites invaded Sulawesi recently and likely in 2 separate invasion events. Our results provide the first records of metakinetoplastids in Sulawesi's rodents and highlight the need for more extensive sampling for pathogens in this biodiversity hotspot., (© American Society of Parasitologists 2020.)

This study investigated the occurrence and phylogenetic relationship of protozoan parasites and Ehrlichia infecting domestic animals from three municipalities in uMkhanyakude district of KwaZulu-Natal province, South Africa. A total of 208 blood samples collected from clinically healthy cattle, sheep, goats and dogs from uMkhanyakude district were examined by polymerase chain reaction (PCR) assays, using either genus or species-specific primers to determine the occurrence and phylogenetic relationship of various protozoan parasites and Ehrlichia of veterinary importance. A total of 5/109 (4.6%) cattle were PCR-positive for the presence of Toxoplasma gondii, 33/109 (30.3%) for Babesia bovis, 24/109 (22.02%) for Babesia bigemina and 20/109 (18.3%) for Trypanosoma sp., while 3/10 (30%) of sheep were PCR-positive for Theileria ovis and none of the goats were positive for any of the detected pathogens. The co-infection of 4/109 (3.7%) B. bovis and B. bigemina was detected in cattle. Only Ehrlichia canis was detected in dogs with infection rate of 20/48 (41.7%). Sequences of PCR-positive isolates (B. bovis, B. bigemina, E. canis, T. ovis and T. gondii) showed that they were closely related to their relevant species from various countries. These findings have expanded our knowledge about the prevalence and phylogenetic similarity between protozoan parasites and Ehrlichia isolates of South African origin. To date, this is the first study in South Africa to detect T. gondii infections from cattle blood using PCR.

This study reports the first autochthonous "surra" outbreak in horses in the State of Santa Catarina, southern Brazil. Six horses with clinical suspicion of trypanosomosis had the natural infection by T. evansi confirmed by PCR and rapid serum agglutination test. Clinical, parasitological, and hematology evaluations were performed at initial observation (T0) and 90 days after (T1). At T0, all animals that tested positive for T. evansi in PCR presented with severe clinical signs and out of normal range hematological hematological (hematocrit, leukocytes, platelets, hemoglobin) and serum biochemical parameters (alanine aminotransferase (ALT), creatinine phosphokinase (CPK), lactate dehydrogenase (LDH), creatinine, bilirubin, and glucose). At T1, the progressive clinical recovery of animals, normalization of hemato-biochemical parameters, and negative PCR results for T. evansi were observed. It was not possible to identify the vector/mechanism of transmission through which animals were infected; therefore, the implementation of surveillance and control measures is essential to prevent the spread of this disease in horse herds, as well as to other animal species., Competing Interests: Declaration of Competing Interest The authors declared no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Detection of trypanosomes in tsetse or domestic livestock is a basic requirement for epidemiological studies as well as for planning and implementing control measures against tsetse and trypanosomiasis. This epidemiological study aimed at assessing the prevalence of trypanosomes in pigs and tsetse flies in the Jomoro district of the western region of Ghana using molecular techniques. Blood was collected from pigs and biconical traps were used to collect tsetse flies. DNA was isolated from 300 pig blood samples and 300 flies for trypanosome detection and identification by PCR. Packed Cell Volume (PCV) of blood samples from 300 pigs was measured using a micro-haematocrit reader. Glossina palpalis palpalis was the only tsetse species found in the area with fly apparent density of 18.4 fly/trap/day. An overall prevalence of trypanosomes in the study area was 4.3% and 0.8% in pigs and tsetse flies respectively. Mixed infection with Trypanosoma (T.) congolense forest and T. vivax was most prevalent 46.2% followed by single infection of T. vivax 15.4%, T. congolense and a mixed infection of T. congolense, T. vivax and T. brucei sl. were the least with 7.7% each. There were no significant differences in trypanosome prevalence among different age groups and between both sexes of the studied pigs (p > 0.05). Trypanosome prevalence was lower in healthy looking 1.9% than the sick looking 20%, pigs (P < 0.05). Mean PCV of parasitaemic pigs 29.3% was significantly lower than that of aparasitaemic pigs 37.8%. Two out of the five species-specific primers used could not identify any trypanosome species from the total blood samples examined. This could possibly mean that those species are not found in the present study area. These results provide useful background information for further study and justification to extend tsetse control to the Jomoro district., Competing Interests: Declaration of Competing Interest All authors declare that they have no Conflict of Interest for the publication of this Manuscript., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The increasing number and severity of surra outbreaks in the Philippines led the government to consider it as the second most important disease of livestock in the country. It is one of the most economically important animal parasitic diseases and has been reported in several animal species, including water buffaloes, cattle, and horses in different regions of the Philippines. However, it has not yet been reported in Cebu, the usual gateway of livestock trade in the area that raises 6% of the 3.75 million goats in the country. In the current study, a PCR-based assay was conducted for the molecular detection and characterization of Trypanosoma evansi in goats in Cebu. A total of 251 goats were randomly sampled from four farms. DNA was extracted and ITS1-PCR was applied to detect different trypanosomes in goats. Eighty-five out of the 251 (33.9%) samples tested positive for T. evansi, two of which were also positive for T. theileri-like trypanosome. The detection rate of T. evansi was slightly higher in male goats (38.3%) than in females (32.5%), and in younger goats (34.5%) than in adults (33.5%). The findings, however, did not differ significantly to suggest any association between sex and age with T. evansi infection in goats. The detection of T. evansi and T. theileri-like trypanosome in goats was confirmed by sequence analysis of ITS1 region. To our knowledge, this is the first report on the molecular detection and identification of caprine T. evansi infection in Cebu, Philippines., Competing Interests: Declaration of Competing Interest The authors declare no competing interests in association with this study., (Copyright © 2020. Published by Elsevier B.V.)

Trypanosoma evansi (T. evansi) is a hemoflagellate parasite that affects a broad range of mammalian hosts and that causes a disease called surra. Diagnosis of surra based on clinical symptoms alone is inaccurate. Therefore, a variety of serological and molecular diagnostic tests are used to assist in the detection of T. evansi infections. The aim of this study was to compare the diagnostic performance of four serological tests (CATT/T.evansi, immune trypanolysis, ELISA with purified variant surface glycoprotein RoTat 1.2 and with whole cell lysate) and two molecular PCR tests targeting sequences within the ribosomal genes locus (ITS1 TD PCR and 18S qPCR). Tests were carried out on blood samples from 161 dromedary camels, 93 horses, 129 goats, 168 sheep, 127 bovines and 76 dogs. Latent class analysis was carried out to calculate the sensitivity and specificity of each diagnostic test. Cohen's Kappa test was used to assess the concordance between the different diagnostic tests. Overall positivity rates observed with the serological tests were as follows: 3.1 % with CATT/T.evansi, 4.9 % with ELISA/RoTat 1.2, 3.4 % with ELISA/whole lysate and 2.0 % with immune trypanolysis (TL). Among the 754 samples tested with the molecular tests, 1.7 % were positive with 18S qPCR and 1.3 % with ITS1 TD PCR. Cohen's Kappa test showed agreement ranging from fair to substantial (k = 0.2-0.8) between serological diagnostic tests. However, it showed a perfect agreement (k = 0.868) between molecular diagnostic tests. Latent class analysis showed that all serological tests were 100 % sensitive, in contrast to the molecular tests with 47 % sensitivity. All tests, though, were highly specific (≥ 97 %). Given the persistence of circulating antibodies after cure, detectable by serological tests, it is recommend combining a serological and a molecular diagnostic test for accurate diagnosis of infection with T. evansi in domestic animals., Competing Interests: Declaration of Competing Interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Callithrix jacchus and C. penicillata marmosets are invasive to the state of Rio de Janeiro, Brazil, threatening the native and vulnerable C. aurita. Both invasive species can be hosts of Trypanosoma cruzi, T. minasense, T. rangeli and T. devei. We aim to investigate the occurrence of trypanosomatids in Callithrix sp. from Jardim Botânico do Rio de Janeiro, located in a central and populous area of the city. Fifteen marmosets were captured. Blood samples were collected for light microscopy and molecular genetics analysis. Parasites morphometric values were evaluated for species identification. DNA was extracted from blood samples by phenol-chloroform method, for partial amplification of the 18S rRNA gene. PCR products were sequenced and aligned using BLAST®. A maximum likelihood phylogenetic tree was constructed to analyze the proximity between the observed sequences. By light microscopy, trypomastigotes were detected in five of the fifteen marmosets. Morphometric measurements and size polymorphism corresponded to those previously described for T. minasense. The DNA sequences of approximately 600 base pairs of the 18S rRNA gene were obtained for three samples with 99% identity with T. minasense sequence, forming a cluster in the phylogenetic tree and corroborating morphometric analysis. Trypanosoma minasense is a highly specific parasite to non-human primates considered as non-pathogenic. There is no evidence of infection in humans and these parasite findings from invasive marmosets do not support additional risks for the native species., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Equine trypanosomiasis is a severe and prevalent disease that has the greatest impact globally upon working equids due to its distribution across lower income countries. Morbidity and mortality rates are high; disease management strategies in endemic regions are ineffective and cost prohibitive. Individual variation in disease phenotype in other species suggests host factors could reveal novel treatment and control targets but has not been investigated in equids., Methods: A prospective clinical evaluation of equines presenting for a free veterinary examination was performed in hyperendemic villages in The Gambia. Age, body condition score and body weight were estimated by validated methods, and haematocrit and total protein concentration measured. Animals fulfilling 2 out of 5 clinical inclusion criteria (anaemia, poor body condition, pyrexia, history of abortion, oedema) for a diagnosis of trypanosomiasis received trypanocidal treatment with follow-up at 1 and 2 weeks. Blood samples underwent PCR analysis with specific Trypanosoma spp. primers and results were compared to the subject's clinical and clinicopathological features. A mixed effects generalised linear model was generated to evaluate the association of infection status with degree of pyrexia and anaemia., Results: Morbidity was high within examined (n = 641) and selected (n = 247) study populations. PCR status was not associated with a defined disease phenotype; there was intra- and inter-species variability. Donkeys were more frequently Trypanosoma spp.-positive (P < 0.001) and febrile (P < 0.001) than horses, but infected horses were more anaemic (P < 0.001), and in poorer body condition (P < 0.001) than donkeys. Sex was correlated to disease phenotype: males were more anaemic (P = 0.03) and febrile (P < 0.001). Haemoparasite co-infections were more common than a single infection., Conclusions: There was evidence of diversity in trypanosomiasis clinical signs plus variable disease phenotypes within equid subpopulations that warrant further investigation. The complex co-infection profile of field cases requires greater consideration to optimise disease management.

Trypanosomes of the subgenus Megatrypanum have been isolated from many mammalian hosts around the world. They are usually non-pathogenic, although they may confuse the parasitological diagnosis of trypanosomosis. Additionally, Trypanosoma theileri has been associated with disease in cattle. Megatrypanum trypanosomes are considered to be transmitted by different arthropods, including tabanids. However, little is known about the potential vectors of Megatrypanum trypanosomes in different parts of the world. The present study reports on the detection of Megatrypanum trypanosomes in Heamatopota pluvialis, Tabanus bromius, Tabanus maculicornis and Tabanus distinguendus in Poland. It also discusses the possible role of these tabanids in the transmission of Megatrypanum trypanosomes., (© 2019 The Royal Entomological Society.)

Background: Animal trypanosomosis caused by Trypanosoma evansi is known as "surra" and is a widespread neglected tropical disease affecting wild and domestic animals mainly in South America, the Middle East, North Africa and Asia. An essential necessity for T. evansi infection control is the availability of reliable and sensitive diagnostic tools. While DNA-based PCR detection techniques meet these criteria, most of them require well-trained and experienced users as well as a laboratory environment allowing correct protocol execution. As an alternative, we developed a recombinase polymerase amplification (RPA) test for Type A T. evansi. The technology uses an isothermal nucleic acid amplification approach that is simple, fast, cost-effective and is suitable for use in minimally equipped laboratories and even field settings., Methodology/principle Findings: An RPA assay targeting the T. evansi RoTat1.2 VSG gene was designed for the DNA-based detection of T. evansi. Comparing post-amplification visualization by agarose gel electrophoresis and a lateral flow (LF) format reveals that the latter displays a higher sensitivity. The RPA-LF assay is specific for RoTat1.2-expressing strains of T. evansi as it does not detect the genomic DNA of other trypanosomatids. Finally, experimental mouse infection trials demonstrate that the T. evansi specific RPA-LF can be employed as a test-of-cure tool., Conclusions/significance: Compared to other DNA-based parasite detection methods (such as PCR and LAMP), the T. evansi RPA-LF (TevRPA-LF) described in this paper is an interesting alternative because of its simple read-out (user-friendly), short execution time (15 minutes), experimental sensitivity of 100 fg purified genomic T. evansi DNA, and ability to be carried out at a moderate, constant temperature (39°C). Therefore, the TevRPA-LF is an interesting tool for the detection of active T. evansi infections., Competing Interests: The authors have declared that no competing interests exist.

Background: Trypanosoma evansi is the causative agent of surra, a disease that occurs in many animal species. The disease is responsible for substantial losses in global production and can be fatal if not diagnosed early. This study aims to determine the prevalence of T. evansi in livestock, equids and dromedary camels in Palestine., Methods: Blood samples were collected during 2015-2017 from domesticated animals (n = 259 animals; 77% females and 23% males) including camels (n = 87), horses (n = 46), donkeys (n = 28), mules (n = 2), sheep (n = 49) and goats (n = 48) from eight districts: Ariha (Jericho), Nablus, Bethlehem, Deir Al Balah, Jenin, Rafah, Tubas, and Khan Yunis. Parasite prevalence was determined using PCR and blood smear microscopy. PCR-positive samples were further phylogenetically analyzed using DNA sequences of the 18S ribosomal RNA gene., Results: The overall infection prevalence was 18% (46/259). The positivity rates according to PCR and microscopy examination were 17% (45/259) and 2.7% (7/259), respectively. The infection rates were as follows: camels, 26/61 (30%); horses, 8/46 (17%); donkeys, 3/28 (11%); mules, 1/2 (50%); sheep, 2/42 (4%); and goats, 6/42 (13%). Phylogenetic analyses of the 18S rRNA gene showed that 24 positive T. evansi samples from Palestine formed a monophyletic cluster with seven T. evansi sequences from Africa, Asia and South America, and three T. brucei sequences from Africa retrieved from GenBank. The spatial analysis showed three statistically significant foci of T. evansi infection in Jenin, Tubas (P = 0.02) and Ariha (Jericho) (P = 0.04). No statistically significant foci were detected in the Gaza Strip., Conclusions: To the best of our knowledge, this is the first confirmation of high levels of infection with T. evansi as a causative agent of surra in Palestine. Our study emphasizes the need for a stringent surveillance system and risk assessment studies as prerequisites for control measures. Further investigations focusing on vectors and evaluation of risk factors are needed.

High-throughput sequencing of cDNA (RNASeq) is now the method of choice for analysis of transcriptomes. This chapter details important considerations in the design of RNASeq experiments for kinetoplastids grown in culture or experimental animals. It contains protocols for obtaining parasites from rodents, and for removal of rRNA from total RNA. In addition, custom pipelines for sequence alignment, and for data analysis and visualization, are described.

Even if the number of Human African Trypanosomiasis (HAT) cases from Kinshasa province in DRC is going towards elimination for the last decade, cases still occur in the periphery of the city. The diagnosis of 21 cases in the south periphery of Kinshasa, between 2015 and 2017 gives evidence of the existence of an active focus in this area. Here, we present the results of a punctual entomological survey that was realized in july 2014 in the outskirts of the southeast of Kinshasa. Using pyramidal traps, we caught tsetse flies during 2 days, dissecting the fresh ones for further molecular analysis. The average Apparent Density of flies per Trap and per Day was three with a maximum of 5.6 flies in Nganda PIO. Polymerase chain reaction analysis of the midguts provided evidence of a high prevalence (57.2%) of infected flies. Ninety three percent of the trypanosomes that were identified belonged to the Nanomonas species, but Trypanozoon trypanosomes were also present in 24% of the infected flies, including mixed infections with Nanomonas, including 3 flies carrying Trypanosoma brucei gambiense, the human pathogen of trypanosomiasis. These results show that at the time of the field's study there was an active reservoir of trypanosomes, closed to pigsties, knowing that pig is a potential animal reservoir. It also demonstrates that xenomonitoring using the entomological approach can be an efficient tool for monitoring sleeping sickness. Finally, results are discussed in the frame of WHO's HAT elimination project. Regarding Kinshasa, it points out the need of regular epidemiologic surveys., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Camel trypanosomiasis or surra is of great concern in Somalia, since the country possesses the largest one-humped camel (Camelus dromedarius) population in the world. Civil war in Somalia has resulted in the destruction of educational, research, economic and social structures, making the country scores very low for most humanitarian indicators. Previous studies on detection of Trypanosoma species in Somali camels have only been performed during the 1990s using standard trypanosome detection methods (STDM). Considering the lack of state-of-the-art knowledge on camel trypanosomiasis in Somalia, the present study aimed to assess the prevalence of Trypanosoma spp. in three districts of Somalia., Methods: A total of 182 blood samples from C. dromedarius from nomadic and dairy farms were evaluated using STDM, serological (CATT/T. evansi) and molecular (ITS1-PCR) methods., Results: All samples were negative for Trypanosoma spp. by STDM. A total of 125/182 (68.7%, 95% CI: 61.4-75.3%) camels were seropositive for T. evansi by CATT/T. evansi. Camels reared in nomadic system were more likely to be seropositive for T. evansi than those under dairy production system (OR: 5.6, 95% CI: 2.1-15.2, P = 0.0001). Five out of 182 (2.7%, 95% CI: 0.9-6.3%) camels tested positive for Trypanosoma sp. by ITS1-PCR. Sequencing of the ITS1 region of the Trypanosoma species detected herein revealed that camels were infected with T. evansi and T. simiae., Conclusions: Trypanosoma evansi is highly prevalent in camels from the Banadir region of Somalia, particularly in nomadic herds. To our knowledge, this is the first study to confirm infections with T. evansi and T. simiae in Somali camels through DNA sequencing. Our data highlight the need for implementation of adequate control measures aiming to reduce the impact on camel production in the country.

Surra, caused by Trypanosoma evansi, is a widely distributed animal trypanosomosis; it affects both domestic and wild mammals with high economic impact. Clinical picture is moderate in bovines but severe in equids. Surra is also an important constraint for international animal trade and movements. Despite its impact, surra remains poorly diagnosed because of low sensitivity tests. To improve epidemiological knowledge of the disease and to secure international movement, efficient diagnosis tools are required. Here, we optimized and applied to equids the OIE-recommended indirect ELISA T. evansi that was validated in other species. Based on 96 positive and 1,382 negative horse reference samples from Thailand, a TG-ROC analysis was conducted to define the cutoff value. ELISA's sensitivity and specificity were estimated at 97.5% and 100%, respectively, qualifying the test to provide a reliable immune status of equids. The test was then applied on 1,961 horse samples from 18 Thai Provinces; the only scarce positives suggested that horses do not constitute a reservoir of T. evansi in Thailand. All samples from racing horses were negative. Conversely, two outbreaks of surra reported to our laboratory, originating from a bovine reservoir, exhibited high morbidity and lethality rates in horses. Finally, posttreatment follow-ups of infected animals allowed us to provide outbreak management guidelines., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2019 Margot Camoin et al.)

Distinct species of Trypanosoma have been documented sharing the same hosts in different environments in intricate transmission networks. Knowing this, this study investigated the role of different hosts in the transmission cycles of Trypanosoma species in the Pantanal biome. The mammals were sampled from November 2015 to October 2016. We sampled a total of 272 wild mammals from 27 species belonging to six orders and 15 families, and three species of triatomines (n = 7). We found high parasitemias by Hemoculture test for Trypanosoma cruzi (TcI), Trypanosoma rangeli, Trypanosoma cruzi marinkellei and Trypanosoma dionisii, and high parasitemias by Microhematocrit Centrifuge Technique for Trypanosoma evansi. The carnivore Nasua nasua is a key host in the transmission cycles since it displayed high parasitemias for T. cruzi, T. evansi and T. rangeli. This is the first report of high parasitemias in Tamandua tetradactyla and cryptic infection in Dasypus novemcinctus by T. cruzi; cryptic infection by T. evansi in Eira barbara, Euphractus sexcinctus and Dasyprocta azarae. The collection of Panstrongylus geniculatus increased the geographic distribution of this vector species in the South America. Our results indicate that Trypanosoma species circulate in a complex reservoir system including different host species with different infective competences., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The prevalence and distribution of Trypanosoma evansi (T. evansi) infection on camels in Somaliland were studied using the card agglutination test (CATT/T. evansi). A total of 2575 camels were surveyed. The overall animal-level apparent (test) prevalence in Somaliland was 26.4%. The prevalence was highest in the Sahil Region (37.3%) followed by Togdheer (28.4%), Sanaag (27.4%), and Sool (26.2%). There was a significant difference in the seroprevalence between the regions (χ 2  = 35.2, df = 1, P < 0.001) with the prevalence being significantly higher in Sahil than in Woqooyi Galbeed and Awdal, respectively. Camels from the eastern districts were two times more likely to be infected than camels from the western districts (P < 0.001). Herds in the coastal area were also more likely to be infected (37.0%) than herds from the inland areas (26.9%).The seroprevalence varied significantly between body condition score groups (P < 0.001) with the highest seroprevalence in camels of poor body condition (39.5%) and the lowest in camels of good body condition (18.4%) score. Animals in poor body condition were almost three times more likely to be positive than animals in good body condition (OR = 2.9; 95% CI 2.2, 3.8).

Trypanosoma evansi is enzootic in camels in Egypt, and water buffaloes act as a reservoir for camel infection. Molecular techniques have contributed towards understanding the epidemiology of T. evansi. Trypanosoma evansi was detected in acute and chronic stages of the disease in male and female mice by polymerase chain reaction (PCR) using two primers. Two experiments were conducted. In experiment I, two groups consisting of 26 female and 26 male mice received 104 trypanosome by I/P inoculation for each mouse. In experiment II, 42 female and 42 male mice were inoculated I/P with 102 trypanosome/mouse. In addition, five mice were kept as uninfected control for each group. Mice were monitored daily for parasitaemia level during the pre-patent period using the micro-haematocrit centrifugation technique (MHCT) and conventional PCR. The primer pairs, (Trypanosoma brucei) TBR1/2 and TeRoTat1.2 (T. evansi Rode Trypanozoon antigen type [RoTat] 1.2), detected the infection after 24 hours earlier than MHCT in both experiments. The course of infection that was detected by MHCT revealed three waves of parasitaemia in female mice and two waves in male mice in the chronic stage of infection. In addition, PCR was able to detect T. evansi in different organs in the chronic stage (i.e. disappearance of parasite from blood). Application of the two primer sets on blood samples from camels showed that all samples were positive by TBR1/2 primers and only 32 of 44 were positive by TeRoTat1.2 primers. Acutely and chronically Trypanosoma-infected mice were detected by PCR in blood and organs. TBR1/2 primers were more sensitive than TeRoTat1.2 primers in detecting Trypanosoma-infected mice, and more reliable in detecting field-infected camels and excluding carrier animals.

The diseases caused by hemoprotozoan parasites in cattle often result in economic losses. In Sri Lanka, previous studies found that the up-country wet zone, which is located in central Sri Lanka, was characterized by a high rate of Theileria orientalis and a low rate of Theileria annulata compared with the dry zone. In this study, DNA samples were prepared from the blood of 121 cattle in Galle, a coastal district located in low-country wet zone in Sri Lanka, and were PCR-screened for Babesia bovis, Babesia bigemina, T. annulata, T. orientalis, and Trypanosoma theileri. All the parasite species, except B. bovis, were detected among the surveyed cattle. The animals had a high rate of T. orientalis (100%) and a low rate of T. annulata (1.6%), as in the up-country wet zone. Babesia bigemina and Tr. theileri were detected in 19.0% and 20.6% of the animals, respectively, and their infection rates were higher in the animals reared in extensive management systems (32.8% and 27.9%, respectively) than in those managed in intensive/semi-intensive systems (5.0% and 13.3%, respectively). Genotypic analyses found that the T. orientalis mpsp type 5 was predominant similar to up-country wet zone, and that Tr. theileri consisted of seven catl genotypes, including two new genotypes (IL and IM) and four previously detected genotypes (IA, IB, II, and IK). These findings suggest that the hemoprotozoan infection profiles are largely conserved within the wet zone, despite differences in the geography, cattle breeds, and management practices between the up-country and low-country wet zones., (Copyright © 2019 Elsevier B.V. All rights reserved.)

From September 2012 to May 2018, blood samples from 364 raptors (mostly adults) were collected and screened for trypanosomes and haemosporidians by microscopic examination and nested polymerase chain reactions (PCR). Trypanosoma spp. were identified in 15 birds from eight different species. Light microscopy revealed 14 cases of infection with Trypanosoma cf. corvi, including one each in black-shouldered kite (Elanus caeruleus, n = 49), Brahminy kite (Haliastur indus, n = 50), and spotted owlet (SO, Athene brama, n = 27); two mountain hawk-eagles (Spizaetus nipalensis, n = 3); and three each in Asian barred owlets (ABO, Glaucidium cuculoides, n = 27), barn owls (BO, Tyto alba, n = 65) and collared scops owls (CSO, Otus lettia, n = 41). In addition, one case of infection with T. avium was identified in an oriental scops owl (OSO, Otus sunia, n = 2). All infected raptors showed very low parasitemia levels. The PCR detected more three positives in one CSO, one Japanese sparrowhawk (Accipiter gularis), and one OSO. The sensitivity and specificity of the PCR method were 93.3% and 99.1%, respectively. The overall infection rate was very low (4.9%). The highest infection rate was recorded in cold-dry season (9.9%). Coinfection of Plasmodium with trypanosomes was found in all three ABOs. Coinfection with Haemoproteus spp. was found in one BO, three CSOs, and one SO. Coinfection with Haemoproteus spp. and Leucocytozoon danilewskyi was found in the OSO. Microfilarias were detected in one ABO and one CSO. The ultrastructure of trypomastigotes of T. cf. corvi in an ABO revealed fine structures. All small subunit ribosomal RNA (SSU rRNA) sequences belong to two clades: T. avium and T. corvi-culicavium complex/group. SSU rRNA gene amplification was not successful in one BO. The raptors with trypanosome infections showed normal hematological values and healthy appearance. Furthermore, this is the first report of T. avium in a nocturnal raptor from Thailand.

We studied infection by Trypanosomatidae in bats captured in two areas with different degradation levels in the Atlantic Forest of Rio de Janeiro state: Reserva Ecológica de Guapiaçu (REGUA) and Estação Fiocruz Mata Atlântica (EFMA). Furthermore, we evaluated whether the diversity of trypanosomatids changes according to bat diversity and the different levels of preservation in the region. The results showed no influence of the level of preservation on bat species richness (15 and 14 species, respectively), with similar chiropterofauna and higher abundance of two common fruit-eating bat species in the tropics: Carollia perspicillata and Artibeus lituratus. Of the 181 bat specimens analyzed by LIT/Schneider hemoculture, we detected 24 infected individuals (13%), including one positive Sturnira lilium individual that was also positive by fresh blood examination. Molecular characterization using nested PCR targeting the 18 SSU rRNA-encoding gene fragment showed similar trypanosomatid infection rates in bats from the two areas: 15% in REGUA and 11% in EFMA (p = 0.46). Trypanosoma dionisii was the most frequently detected parasite (54%), followed by T. cruzi DTUs TcI and TcIV and Trypanosoma sp., in Neotropical phyllostomid bats (RNMO63 and RNMO56); mixed infections by T. dionisii/T. cruzi TcIII and T. dionisii/T. cruzi TcI were also observed. The T. cruzi DTUs TcI and TcIV are the genotypes currently involved in cases of acute Chagas disease in Brazil, and T. dionisii was recently found in the heart tissue of an infected child. Surprisingly, we also describe for the first time Crithidia mellificae, a putative monoxenous parasite from insects, infecting a vertebrate host in the Americas. Bats from the Atlantic Forest of Rio de Janeiro state harbor a great diversity of trypanosomatids, maintaining trypanosomatid diversity in this sylvatic environment., Competing Interests: The authors have declared that no competing interests exist.

A laboratory prototype system that correlates murine blood absorbance with degree of infection for Plasmodium berghei and Trypanosoma avensi has been designed, constructed and tested. A population (n = 6) of control uninfected, Plasmodium infected and Trypanosoma infected BALB/c mice were developed and spectral absorption measurements pre and post infection were made every 3 days. A fibre optic spectrometer set-up was used as the basis of a laboratory prototype biosensor that uses the Beer Lambert Law to relate Ultraviolet-Visible-Near-infrared absorbance data to changes in murine blood chemistry post infection. Spectral absorption results indicate a statistically relevant correlation at a 650 nm with infection for Plasmodium from between 4 and 7 sampling days' post infection, in spite of significant standard deviations among the sample populations for control and infected mice. No significant spectral absorption change for Trypanosoma infection was been detected from the current data. Corresponding stained slides of control and infected blood at each sampling date were taken with related infected cell counts determined and these correlate well for Plasmodium absorbance at 650 nm., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The aim of this study was to evaluate the effect of trypanosomes on cultured largemouth bass (Micropterus salmoides) and describe the taxonomic identification of the parasite. The effects of the parasite on M. salmoides were examined based on clinical symptoms, hemograms, histopathology, and serum biochemistry. Diseased fish showed typical clinical symptoms of trypanosomiasis, which included lethargy, anorexia, and histopathological lesions in the liver, head kidney, and spleen. The serum of diseased fish had significantly lower concentrations of glucose, triglyceride, and low-density lipoprotein, and significantly higher alanine transaminase (ALT), aspartate transaminase (AST), and lactate dehydrogenase (LDH) activities. The morphology of the trypanosomes was also analyzed using light microscopy, and their 18S rDNA sequence was analyzed to establish genetic relationships with other known strains. We found that the trypomastigote form of the trypanosomes from M. salmoides was similar to those isolated from Pelteobagrus fulvidraco. The trypanosomes had a slender and narrow body with a relatively long free flagellum, not well-developed undulating membrane, and an oval kinetoplast located near the subterminal posterior end of the body. The 18S rDNA sequences of the trypanosome from M. salmoides had the highest similarity (99.8%) with that of P. fulvidraco, suggesting they are identical species. Based on the differences in morphological characteristics and 18S rDNA sequence compared to trypanosomes isolated from other freshwater fish, it is considered as a new species and we propose the name Trypanosoma micropteri n. sp.

Trypanosoma (Megatrypanum) theileri is a cosmopolitan, usually non-pathogenic, trypanosome of cattle transmitted by blood-sucking arthropods, mainly tabanid flies. Several T. theileri strains isolated from domestic and wild ruminants via co-culturing with mammalian feeder cells or blood cells have been characterized morphologically and genetically. Here, we cultured a new trypanosome isolate from a Holstein cow in Hokkaido, Japan, and performed morphological and molecular characterization studies. The new isolate (Obihiro strain) was co-cultivated with Madin-Darby bovine kidney (MDBK) cells in GIT medium supplemented with 10% fetal bovine serum. Trypomastigotes and epimastigotes, but not intracellular parasites, were identified in the culture. Analysis of the V7-V8 region of 18S rRNA sequences showed that the Obihiro strain is positioned within the subgenus Megatrypanum. A dendrogram based on whole internal transcribed spacer rDNA sequence showed that the Obihiro strain clustered in the lineage TthII together with the Japanese isolates of T. theileri, Esashi 9, and Esashi 12, and isolates from Zambia and the USA. T. theileri of the KM strain and a T. theileri-like trypanosome isolated from deer (TSD1 strain) clustered in the lineage TthI, separate from the Obihiro strain. Based on a partial cathepsin L-like protein gene analysis, the Obihiro strain clustered with isolates of the TthIIF genotype, which includes T. theileri from Vietnam, Sri Lanka, and Brazil. Our analyses of the T. theileri Obihiro strain provide relevant insights into its genetic diversity in Japanese cattle and corroborate the host specificity of cattle and deer trypanosomes of the subgenus Megatrypanum.

The interactions of host, vector and parasite in bovine trypanosomiasis transmission cycles in southwest Nigeria are not yet well understood. Trypanosoma (Trypanosomatida: Trypanosomatidae) species infection prevalences and bloodmeal sources were determined in transmitting vectors of the genera Glossina (Diptera: Glossinidae), Tabanus (Diptera: Tabanidae) and Stomoxys (Diptera: Muscidae) collected using Nzi traps in cattle settlements in southwest Nigeria. Sequenced cytochrome B mitochondrial DNA segments obtained from vector digestive tracts identified bloodmeal sources from eight host species, namely human, cattle, hippopotamus, giraffe, gazelle, spotted hyena, long-tailed rat and one unidentified species. Overall, 71.1% [95% confidence interval (CI) 63.0-78.1], 33.3% (95% CI 21.9-47.0) and 22.2% (95% CI 16.2-29.9), respectively, of Glossina, Tabanus and Stomoxys flies were positive for trypanosomes. The observed trypanosome species were Trypanosoma vivax, Trypanosoma congolense, Trypanosoma brucei, Trypanosoma evansi, Trypanosoma simiae and Trypanosoma godfreyi. Trypanosome DNA was more prevalent in tsetse (34.8% Tr. vivax, 51.1% Tr. b. brucei, 5.2% Tr. congolense, 4.4% Tr. simiae and 24.4% mixed infections) than in other flies and the main determinants in all flies were seasonal factors and host availability. To the best of the present group's knowledge, this is the first report of Trypanosoma species in Tabanus and Stomoxys flies in Nigeria. It indicates that vector control programmes should always consider biting flies along with tsetse flies in the control of human and animal trypanosomiasis., (© 2019 The Royal Entomological Society.)

Although bats were considered as a major host of trypanosomatid flagellates, information of trypanosomes in bats is unknown in China. We collected bats in 2015 from Shandong Province of China and used PCR to amplify the Trypanosoma glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) gene and 18S rRNA gene from the bat blood samples and heart tissues. The results showed that 10.3% (13/126) of bats (Eptesicus serotinus and Myotis pequinius) were positive for trypanosomatid DNA and DNA sequencing showed that all PCR amplified Trypanosoma DNA belonged to T. dionisii. We concluded that T. dionisii had a infection rate in bats from China. For the first time, Trypanosoma infections were detected in bats from China, providing valuable information on the prevalence of these parasites in Asia. This is also the first report of Trypanosoma dionisii in Myotis pequinius, suggesting that Trypanosoma dionisii has a broad host species., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Surra is an animal trypanosomosis, caused by infection with Trypanosoma evansi and leading to severe economic loss due to mortality and morbidity. Compared to tsetse-transmitted animal trypanosomoses, little attention is given to the epidemiology and control of surra. Understanding its epidemiology is a first step in local and global efforts to control the disease. We conducted a systematic review and meta-analysis of published studies on distribution, host ranges and prevalence of T. evansi infection., Methods: Four electronic databases were searched for publications on T. evansi that met our inclusion criteria for the systematic review. Subsets of publications were subjected to meta-analysis for the pooled prevalence of T. evansi in various hosts as determined by multiple detection methods., Results: A total of 272 references published between 1906-2017 were included. Trypanosoma evansi was reported from 48 countries; largely confined to Africa and Asia with publications on natural T. evansi infections from 77% (n = 48) of countries, contrasting with seven countries in South America, and four in Europe where T. evansi is not endemic but was imported with infected animals. Although surra is a notifiable disease, many countries do not report surra cases to OIE. Trypanosoma evansi was mainly reported from dromedary camels in Africa and the Middle East, water buffaloes, cattle, dogs and horses in East and Southeast Asia. In South America, the acute form of the disease was reported in horses and dogs. Surra was also reported in a wide range of wild animals. Some rare human cases occurred in India and Vietnam. Meta-analysis on a subset of 165 publications indicated pooled prevalence of T. evansi in domestic animals ranging from 14-31%, 6-28% and 2-9% using respectively antibody detection, molecular and parasitological tests, with camels as the most affected, followed by buffalo and cattle., Conclusions: This study illustrates that T. evansi affects a wide range of domestic and wild animals in Africa, Asia and South America with highest prevalence observed in dromedary camels. For successful control of T. evansi, both locally and globally, the role of wild animals in the epidemiology of surra needs further investigation.

Trypanosoma lewisi (Kinetoplastea: Trypanosomatida: Trypanosomatidae) with a cosmopolitan distribution is the type species of the subgenus Herpetosoma, which includes ca. 50 nominal species isolated mainly from rodents. Since members of Herpetosoma in different host species have an almost identical morphology of bloodstream forms, these trypanosomes are referred to as 'T. lewisi-like', and the molecular genetic characterization of each species is necessary to verify their taxonomy. In the present study, we collected blood samples from 89 murid rodents of 15 species and 11 soricids of four species in Indonesia, Philippines, Vietnam, Taiwan, and mainland China for the detection of hemoprotozoan infection. T. lewisi and T. lewisi-like trypanosomes were found in the blood smears of 10 murid animals, which included Bandicota indica (two rats), Rattus argentiventer (one rat), and Rattus tiomanicus (two rats) in Indonesia; Rattus rattus (one rat) in the Philippines; and Niviventer confucianus (four rats) in mainland China. Furthermore, large- or medium-sized non-T. lewisi-like trypanosomes were detected in two soricids, Crocidura dracula in Vietnam and Anourosorex yamashinai in Taiwan, respectively. Molecular genetic characterization of the small subunit (SSU) ribosomal RNA gene (rDNA) and glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene indicated that the trypanosomes from all the murid hosts had identical SSU rDNA or gGAPDH gene nucleotide sequences except for those in N. confucianus in mainland China. These N. confucianus-infecting trypanosomes also showed several unique morphological features such as smaller bodies, anteriorly positioned nuclei, and larger rod-shaped kinetoplasts when compared with T. lewisi trypomastigotes. Trypanosoma (Herpetosoma) niviventerae n. sp. is erected for this new species. Similarly, based on morphological and molecular genetic characterization, Trypanosoma sapaensis n. sp. and Trypanosoma anourosoricis n. sp. are proposed for the trypanosomes in C. dracula in Vietnam and A. yamashinai in Taiwan, respectively. More effort directed toward the morphological and molecular genetic characterization of the trypanosomes of rodents and soricids is required to fully understand the real biodiversity of their hemoflagellates.

We conducted a cross-sectional study to detect trypanosome infections of horses and donkeys in the Riyadh Province of Saudi Arabia. DNA was extracted from blood samples collected from 368 horses and 142 donkeys, and subjected to universal first ribosomal internal transcribed spacer region (ITS1)-PCR followed by Trypanosoma evansi species-specific RoTat1.2-PCR. The universal ITS1-PCR revealed T. evansi infection in horses ( n = 12; 3.3%) and donkeys ( n = 4; 2.8%). There was no significant effect of sex or age on the prevalence of trypanosomiasis in horses or donkeys. Application of the RoTat1.2-PCR revealed that the RoTat1.2 VSG gene was absent from the positive ITS1-PCR samples of 3 horses and 1 donkey. This discrepancy could be explained by the circulation of T. evansi type B in Saudi Arabia; however, this suspicion requires confirmation.

In Egypt, although the Trypanosoma evansi has been reported frequently among domestic animals, there is no published data on T. evansi in horses. Therefore, this study aimed to assess the prevalence and characterization of T. evansi in three governorates by examining blood samples from 40 local camels, 35 imported camels, 25 horses and 10 donkeys by PCR targeting the sequences of TBR and RoTat 1.2VSG. The overall prevalence of T. evansi was 54.5% and 21.8% by TBR PCR and RoTat 1.2VSG PCR, respectively. The TBR PCR detected T. evansi in 60% and 71.4%, respectively, of local and imported camels and in 10% and 40% of donkeys and horses, respectively. For RoTat 1.2VSG PCR, T. evansi was detected in 32.5% and 31.4 of local and imported camels, respectively. All horses and donkeys were negative by RoTat 1.2VSG PCR. TBR PCR was superior to RoTat 1.2VSG PCR in T. evansi infection detection. Statistically significant differences in the prevalence of the infection were observed on the basis of body condition and location. Sequencing and phylogenetic analysis, based on RoTat 1.2VSG, confirmed the presence of T. evansi, which was closely related to Egyptian and Indian isolates. In conclusion, TBR PCR is the best assay to monitor T. evansi infections in camels, horses, and donkeys. The presence of T. evansi in horses and donkeys possibly play a role in the transport of the infection to camels. This is the first report of T. evansi infection in horses in Egypt using TBR PCR., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Hemoparasitic diseases like trypanosomiasis have an adverse influence on the health and working capability of infected animals. Monitoring and identification of blood born parasitic infections in dairy animals are of vital importance to get the optimum production. In this study blood samples were collected from Nili Ravi buffaloes (n = 390) kept at different villages of district Lodhran, Punjab province of Pakistan. Blood samples were evaluated for red blood cell counts, total and differential leukocyte counts, hematocrit, hemoglobin, total proteins and different serum parameters such as alanine aminotransferase, aspartate aminotransferase, malondialdehyde, alkaline phosphatase, lactate dehydrogenase, phosphorous, copper, calcium and magnesium. Overall prevalence of Trypanosoma evansi was 4.61% based on microscopic smear examination, 11.02% with Formol Gel Test and 16.15% with PCR. Infected buffaloes showed different clinical signs, including high fever (105 ± 1.0 °F), edema of face and legs, hyperemic mucosa of eyes, lachrymation, bulging eyes, pale mucus membranes and frequent urination. Microscopic examination of blood films showed morphologically different parasites. Statistical analysis did not indicate an association of infection based on age and sex of buffaloes. Results revealed significantly (p < 0.05) lower values of red blood cell counts, hemoglobin, hematocrit, mean corpuscular hemoglobin concentration and total proteins, while increased values of mean corpuscular volume, total white blood cells, monocyte, neutrophils, basophils and eosinophils in infected animals. Infected buffaloes were suffering from macrocytic hypochromic anemia. A significant (p < 0.05) increase in serum lipid per oxidation product (malondialdehyde) level and serum enzymes while a decrease in macrominerals and trace mineral (copper) in trypanosomiasis positive buffaloes were recorded. It was concluded that Trypanosoma evansi is prevalent in Pakistan under tropical and subtropical climatic conditions. It causes clinical disease with macrocytic hypochromic anemia and oxidative stress in infected buffaloes., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Trypanosoma evansi is the most widespread of the pathogenic salivarian trypanosomes; it causes a serious disease called surra that affects domestic animals such as camels, horses, and dogs, and often leads to reduced productivity and economic losses. Therefore, the objectives of the present study were to determine the prevalence rates of trypanosomiasis using 3 parasitological tests (wet blood film, Giemsa staining, and microhematocrit centrifugation technique) and polymerase chain reaction (PCR) among stray dogs from Riyadh Province, Saudi Arabia. In the current study, 117 dog blood samples collected from certain districts of Riyadh Province showed that 5 of 117 dogs (4.3%) were positive for the genus Trypanosoma. In addition, the findings indicated no effect of dog gender or age on parasite infection. For a more specific diagnosis, PCR amplification of the RoTat 1.2 VSG gene in 5 internal transcribed spacer1-positive samples diagnosed with Trypanosoma indicated that 2 were positive for RoTat 1.2 T. evansi. The absence of the RoTat 1.2 VSG gene in 3 of the 5 T. evansi-positive samples could be explained by the circulation of T. evansi type B in dogs from Saudi Arabia. Thus, this is the first study demonstrating T. evansi type B outside of Africa.