Trypanosome Species Research Articles

First detection and phylogenetic analysis of Trypanosoma species in European wolves and bears: discovery of novel haplotypes.

Wild carnivores may be important sources of emerging zoonotic pathogens worldwide, however, specific data concerning their presence in large carnivores are limited. The genus Trypanosoma (protozoan parasites) comprises numerous species adapted to all classes of vertebrates. This genus includes highly pathogenic species, such as T. brucei ssp., T. congolense or T. vivax. The aim of this study was to expand on the ecological or zoonotic significance of detecting Trypanosoma in large carnivores. Samples from free-living carnivores (n = 26) were collected. The diversity and occurrence of Trypanosoma species among wildlife in Europe were investigated through the analysis of 18S rRNA gene sequences. The phylogenetic analysis showed three new haplotypes of the 18S rRNA gene of Trypanosoma sp. isolated from two grey wolves (Canis lupus) and two brown bears (Ursus arctos). To the best of our knowledge, this this study represents the first detection of trypanosomes in large carnivores in Europe. The detection of Trypanosoma sp. was achieved through nested- PCR amplification. Furthermore, the identification of new haplotypes of these protozoan highlights variability within Trypanosoma sp., which could lead to the emergence of new, potentially more virulent variants of these parasites in wild animal populations, posing a potential threat to their health. Moreover, these findings underscore the need for further research to comprehensively understand the diversity, distribution, and potential impact of Trypanosoma species on the populations of large carnivores.

Morphological and molecular characterization of avian trypanosomes in domestic chickens (Gallus gallus domesticus) in Southeast Asia and review of the parasite morphometry in different avian hosts

ABSTRACT Avian trypanosomiasis is caused by flagellate protists of the genus Trypanosoma (Trypanosomatidae, Kinetoplastida). These parasites are transmitted by various blood-sucking arthropods. While these parasites are generally considered to have low pathogenicity, certain species can induce clinical symptoms in susceptible birds. In this study, detail morphological and molecular characteristics of Trypanosoma species isolated from domestic chicken (Gallus gallus domesticus) in Northeastern Thailand were investigated. In total, 58 buffy coat blood films, 60 thin blood films of fresh blood and 60 partial DNA sequences were analysed. Subsequently, Trypanosoma avium and Trypanosoma calmettei, which are rarely reported species in poultry as well as Trypanosoma sp. were characterised. Additionally, we provided the details of reviewed information of the parasite morphometry in different avian hosts. Phylogenetic analysis identified the most closely related avian Trypanosoma lineages. This study reports T. avium in domestic chickens and along with molecular characterization of T. calmettei, which are rarely reported species. Buffy coat and nested-PCR diagnostic methods were nearly similar in terms of sensitivity for trypanosome diagnostics, and both methods were more sensitive in comparison to microscopic examination of thin blood films. Relatively cheap buffy coat method is helpful for screening of trypanosomes infection in poultry, particularly during rapid fieldwork. In parasitology research, buffy coat and nested-PCR diagnostics are worth combining with thin blood smear microscopic examination, which provides valuable information about non-deformed trypomastigotes that are essential for morphospecies identification.

Prevalence of <i>Trypanasomiasis</i> in Northern Nigeria: A review

Trypanosomiasis, a parasitic disease caused by protozoan parasites of the genus Trypanosoma, is a significant public health concern in sub-Saharan Africa, including Northern Nigeria. The disease, transmitted by tsetse flies, poses a significant threat to human health, livestock production, and economic development. Animal African trypanosomiasis (AAT), also known as Nagana, is caused by various trypanosome species, including T. brucei, T. congolense, T. vivax, T. equiperdum, T. evansi, T. simiae, T. suis, and T. theileri. Cattle are the most affected, but other animals such as goats, dogs, sheep, pigs, and wild animals are also susceptible. Symptoms of AAT include parasitaemia, anemia, loss of weight, reduced productivity, and high mortality, which limit the pace of rural development in tropical Africa. The current threat of AAT ranks among the top cattle diseases on sustainable livestock production and mixed farming. At least 3 million livestock deaths each year are caused by AAT, with a loss in livestock production and mixed agriculture valued at $5 billion US dollars yearly in Africa. Effective and sustainable control measures can result in up to threefold increases in the estimated livestock population in Nigeria. Trypanosomiasis is considered a haemoparasites, posing a major threat for the development of the cattle industry in Africa, Asia, and Latin America. It kills a variety of domestic and wild animals and causes anemia, weight loss, miscarriage, and productivity losses. In Northern Nigeria, both human African trypanosomiasis (HAT) and animal African trypanosomiasis (AAT) are prevalent. The epidemiology of trypanosomosis depends on the distribution of vectors, the virulence of the parasite, and the host's response.

Molecular detection of Trypanosoma minasense in captive black-faced black spider monkeys Ateles chamek (mammalia; primate)

The present study aimed to assess Trypanosoma spp. infection in 17 captive black-faced black spider monkeys (Ateles chamek), living at the Zoological Garden of Curitiba, Paraná, Brazil. Blood samples (3 mL) were collected by femoral puncture by certified veterinarians. A total of 5 μL of extracted blood DNA was submitted to Nested-PCR 18S rDNA. In overall, 4/17 (23.5%) spider monkeys were positive to Trypanosoma spp. by PCR. Sequencing results for 18S rDNA revealed Trypanosoma minasense infection. This is the first report of T. minasense in black-faced black spider monkey worldwide and the first assessment of Trypanosoma species in NHP of southern Brazil.

Study of bovine trypanosomiasis in Mali: Case of the Kita region

This study conducted between September and December 2022 in six municipalities in the Kita circle, with the aim of determining the prevalence of bovine trypanosomiasis. A sample of 450 cattle had their blood taken to test for trypanosomes. The cattle examined consisted of 239 females and 211 males. The different species of trypanosomes identified using the Buffy coat technique. This technique made it possible to obtain a prevalence of trypanosomes of 22.67%. This prevalence varied depending on sex (male 25.12% and female 20.50%), age (young 33.05% and adults 11.21%) and the species of trypanosomes (Trypanosoma. congolense 61.76%, T. vivax 26.47% and T. brucei 11.76%).

Prevalence of bovine trypanosomiasis in the kangaba circle (Mali)

This study carried out between September and November 2023, in the Kangaba circle aimed to determine the prevalence of trypanosomiasis. Blood samples taken from 219 cattle comprising 142 females and 77 males examined for trypanosomes. Trypanosomes identified using the Buffy coat technique. This work made it possible to obtain a prevalence of 35.16% bovine trypanosoma. Statistical analysis revealed a statistically significant difference (p<0.05) between the prevalence rates observed in the different species of trypanosomes identified, such as Trypanosoma .congolense (68.83%), T. vivax (19.48%). %) and T. brucei (11.69%). The prevalence of trypanosomiasis observed in male and female cattle was 42.86% for male cattle and 30.99% for females. Statistical analysis of the results revealed a significant difference (p<0.05) between trypanosomiasis prevalence rates observed in adult cattle (8.14%) and in young cattle (52.63%). In the commune of Kaniogo, out of 110 cattle examined, 42 individuals were positive for trypanosomiasis, representing an overall prevalence rate of 38.18%. Regarding the commune of Karan, 35 cattle were positive for trypanosomiasis out of 109 subjects examined, representing an overall prevalence rate of 32.11%. The results of our work revealed that bovine trypanosomosis constitutes a real threat to cattle in the Kangaba circle.

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

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

Animal Trypanosomiasis: Challenges and Prospects for New Vaccination Strategies.

Animal trypanosomiasis, such as nagana, surra, and dourine, represent a significant challenge to animal health and economic development, especially in tropical and subtropical regions where livestock production is an essential component of a country's economy. Despite advances in the control of human trypanosomiasis, animal diseases caused by several species of trypanosomes remain neglected. The lack of funding for the development of new treatments and vaccines contributes to sustaining the severe economic impacts these diseases have on the farming industry, especially in low-income rural areas. Recent advances in the understanding of the immune processes involved during infection have been essential for the development of new approaches towards disease control including vaccines. These new approaches must be part of integrated control programs, which must also include vector management and the awareness of good veterinary practices. Addressing the challenges posed by the control of animal trypanosomiasis requires collaborative and continuous efforts shared among scientists, governments, and the farming industry, if significant progress is to be made to mitigate the impact of these diseases. In this literature review, we discuss the main challenges for the development of vaccines for animal trypanosomiasis and the research underway, including the prospects for employing new vaccine platforms, such as an mRNA vaccine, vector-based vaccine, and CRISPR-attenuated parasite vaccine.

Discovery and Development of an Advanced Lead for the Treatment of African Trypanosomiasis.

African trypanosomiasis is a widespread disease of human and veterinary importance caused by various Trypanosoma spp. with a globally devastating impact and a need for novel treatment options. We here provide a comprehensive preclinical evaluation of nucleoside analogues, 6-thioether-modified tubercidins, with curative activity against African trypanosomiasis. Promising hits were identified following in vitro screening against the most relevant trypanosome species. Selected hit compounds were extensively tested for in vitro metabolic stability, potency in in vivo mouse models for the various species, genotoxicity in an in vitro testing battery, and mode of action studies (i.e., genome-wide RNA interference library screening and metabolomics). Among the nucleoside analogues, analogue 3 was curative in mouse models with no indication of genotoxicity and a low ecotoxicological footprint. Mode-of-action studies revealed that P1-type nucleoside transporters and adenosine kinase are involved in the uptake and activation, respectively. Analogue 3 represents a potent, advanced lead fitting the preferred target product profile for a broad-spectrum trypanocide regardless of the causative species.

First report of Trypanosoma microti in Cricetid rodents from Querétaro, Mexico: Expanding the known distribution of Herpetosoma in the Mexican transition zone

Members of the subgenus Herpetosoma are specialized parasites of mammals of the order Lagomorpha and Rodentia. It is estimated that around 50 species within this subgenus are transmitted by several flea species. These parasites have recently gained attention from a public health perspective because several species have been reported to infect humans. Several species of the Herpetosoma subgenus have been recognized on the American continent, although there have been no reports from the Mexican Transition Zone. Therefore, the present study aimed to report the presence of a member of the Herpetosoma subgenus in cricetid rodents from Querétaro, Mexico.A total of 17 rodents belonging to three species, Peromyscus boylii, Peromyscus gratus, and Peromyscus melanophrys, were collected. Genetic material was extracted using the Chelex 100 resin, followed by PCR and sequencing of a fragment of 800 bp of the 18S ribosomal gene of the genus Trypanosoma. Out of these hosts, eight individuals of P. gratus (1♀), P. melanophrys (1♂), and P. boylii (3♂, 3♀) tested positive for Trypanosoma microti DNA, previously detected in the England and Japan. The present study, therefore, provides the first discovery of T. microti in Mexico, thereby increasing the known richness of species of the genus Trypanosoma in rodents from this country and the Mexican Transition Zone.

Insights into trypanosomiasis transmission: Age, infection rates, and bloodmeal analysis of Glossina fuscipes fuscipes in N.W. Uganda.

Tsetse flies (Glossina) transmit species of Trypanosoma which cause human African trypanosomiasis (HAT) and animal African trypanosomiasis (AAT). Understanding the epidemiology of this disease and controlling the vector rationally requires analysis of the abundance, age structure, infection rates and feeding patterns of tsetse populations. We analysed a population of G. fuscipes fuscipes in the Koboko district of Uganda. Seasonal variation in the abundance of tsetse was assessed from the numbers of tsetse caught in pyramidal traps. The age structure of the population was assessed by dissecting female tsetse to estimate their ovarian categories. Classical and PCR-based methods were utilised to determine the presence of the three major pathogenic species of salivarian trypanosomes: T. vivax, T. congolense and T. brucei in a subset (n = 2369) of flies. Further, bloodmeal analysis was carried out using PCR to amplify and sequence a portion of the vertebrate cytb gene. The abundance and age structure of tsetse populations were relatively stable and a slight seasonal four-fold variation in abundance appeared to be correlated with rainfall. Analyses of age structure suggests a low natural daily mortality of 1.75% (1.62-1.88). Infection rates estimated were significantly greater (1.9-9.3 times) using the PCR-based method compared to the classical dissection-based method. Positive rates for T. brucei sl, T. congolense and T. vivax were 1.6% (1.32-2.24), 2.4% (1.83-3.11and 2.0% (1.46-2.63), respectively by PCR. The majority of bloodmeals were identified as cattle (39%, 30.5-47.8) and human (37%, 28.4-45.6). The seasonally stable abundance, low mortality rate and high proportion of bloodmeals from humans may explain, in part, why this district has historically been a focus of sleeping sickness. Additionally, the high rates of cattle feeding indicate insecticide treated cattle may prove to be a useful vector control strategy in the area.

Design, synthesis, in vitro and in vivo trypanosomaticidal efficacy of novel 5-nitroindolylazines

Leishmaniasis and trypanosomiasis rank among lethal vector-borne parasitic diseases that are endemic in tropical and sub-tropical countries. There are currently no preventive vaccines against them, and once diagnosed, a handful of less effective drugs clinically accessible are the only therapeutic options offered to treat these ailments. And although curable, the eradication and elimination of these diseases are hampered by the emergence of multidrug-resistant strains of the causal pathogens. Hence, there is accrued necessity for the development of new, effective, and affordable drugs. In recent decades, several molecular scaffolds, including nitroaromatics, endoperoxides, etc., have been attempted as building blocks to generate new effective clinical antitrypanosomatid agents with low toxicity so far to no avail. In this regard, a series of nitroindolylazine derivatives was synthesised in a three-step process involving nucleophilic substitution (SN), hydrazination and Schiff base condensation reactions, and was evaluated against various Leishmania and Trypanosoma species and strains. Several promising hits portraying leishmanicidal and trypanocidal with in vitro submicromolar activities, and devoid of toxicity on mammalian cells were uncovered. Among these, nitrofurylazine 11 (Tc IC50: 0.08 ± 0.03 μM) and nitrothienylazine 13 (Tc IC50: 0.09 ± 0.01 μM) were evaluated in vivo against Trypanosoma congolense, the causative agent of nagana, which is livestock most virulent trypanosome species in mice-infected preliminary study. However, only partial efficacy was observed as all mice succumbed due to high parasitemia within 13 days post-infection during the treatment. The translational potential of antileishmanial and antichagasic hits, as well as further identification of their molecular targets, will be assessed in future research.

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

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

Detection of tick-borne pathogens in the pangolin tick, Amblyomma javanense, from Vietnam and Laos, including a novel species of Trypanosoma

Two species of Southeast Asian pangolins (the Chinese pangolin, Manis pentadactyla and the Malayan or Sunda pangolin, Manis javanica) are critically endangered species. Therefore, knowledge on their parasitic infections is very important, especially considering ticks that can transmit which pathogens. In this study, 32 pangolin ticks (Amblyomma javanense), that were collected in Vietnam and Laos, were analyzed with molecular methods for the presence of tick-borne pathogens. Two members of the family Anaplasmataceae were shown to be present in 14 pangolin ticks, i.e., Candidatus Anaplasma pangolinii and an Ehrlichia sp. In three ticks, a single Rickettsia genotype was also detected, and in seven ticks four 18S rRNA sequence variants of a Babesia sp. Most importantly, a novel protozoan agent, tentatively called here Trypanosoma sp. “PAT14” was detected in one A. javanense nymph. These results imply the first molecular finding of any species of Anaplasma, Ehrlichia and Babesia in pangolin ticks from Vietnam and Laos. On the other hand, detection of a new tick-associated Trypanosoma sp. in A. javanense from Southeast Asia is not only important from a taxonomic point of view, but it is also the first finding of any trypanosomes in the genus Amblyomma in Eurasia and adds pangolins to the potential placental mammalian hosts of any trypanosomes.

Domestic dogs as reservoirs for African trypanosomiasis in Mambwe district, eastern Zambia

The control of African trypanosomiasis (AT) in Eastern and Southern Africa, including Zambia, faces huge challenges due to the involvement of wild and domestic animal reservoirs. Free-roaming dogs in wildlife-populated and tsetse-infested villages of Zambia’s Mambwe district are exposed to infectious tsetse bites. Consuming fresh raw game meat and bones further exacerbates their risk of contracting AT. We focus on the reservoir role of such dogs in maintaining and transmitting diverse species of trypanosomes that are infective to humans and livestock in Zambia’s Mambwe district. A cohort of 162 dogs was enrolled for follow-up at 3 different time points from June to December 2018 in selected villages of Malama, Mnkhanya, and Nsefu chiefdoms of Mambwe district, eastern Zambia. Blood and serum were screened for AT by microscopy, GM6 ELISA, PCR (ITS1 and SRA), and Sanger sequencing. Out of the 162 dogs in the cohort, 40 were lost to follow-up and only 122 remained traceable at the end of the study. GM6 ELISA detected Trypanosoma antibodies in 121 dogs (74.7%) and ITS1-PCR detected DNA involving single and mixed infections of T. congolense, T. brucei, and suspected T. simiae or T. godfreyi in 115 dogs (70.9%). The human-infective T. b. rhodesiense was detected by SRA PCR in 67 dogs (41.4%), and some sequence data that support the findings of this study have been deposited in the GenBank under accession numbers OL961811, OL961812, and OL961813. Our study demonstrates that the Trypanosoma reservoir community in Zambia is wider than was thought and includes domesticated dogs. As dogs are active carriers of human and livestock-infective trypanosomes, they pose a risk of transmitting AT in endemic villages of Mambwe district as they are neglected and left untreated. To fully bring AT under control, countries such as Zambia where the role of animal reservoirs is important, should not limit their prevention and treatment efforts to livestock (especially cattle) but also include dogs that play an integral part in most rural communities.

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

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

Revisiting the diversity and phylogenetic relationships of trypanosomes (Trypanosoma) infecting pelomedusid (Pelomedusidae) freshwater turtles in Southern Africa

Abstract Species of Trypanosoma infecting reptiles are poorly understood and understudied. The study of trypanosomes infecting turtles could lead to a greater understanding of the genus and its evolutionary history. The present study re-evaluates and classifies two previously described species of Trypanosoma in freshwater pelomedusid turtles, Trypanosoma (Haematomonas) neitzi and Trypanosoma (Haematomonas) sheppardi, and characterizes a new species, Trypanosoma (Haematomonas) pienaari, using morphological and molecular data. This study provides the first morphological descriptions and molecular data of South African turtle trypanosomes, setting a base for future research of the reptile trypanosomes of Africa.

Prevalence of trypanosomiasis caused by Trypanosoma evansi (Kinetoplastea, Trypanosomatidae) in domestic ruminants from Southern Punjab, Pakistan.

Trypanosomiasis, a parasitic infection caused by various Trypanosoma species, poses a significant threat to global livestock, affecting both human health and economic sectors. This study aimed to estimate the prevalence of Trypanosoma evansi in Southern Punjab, Pakistan, focusing on key ruminant species, including camels, cattle, buffaloes, goats, and sheep. A total of 240 blood samples, comprising 48 samples from each animal species (camel, cattle, buffaloes, goat, and sheep) were collected from three districts in Southern Punjab. The collected samples were subjected to thin smear microscopy, DNA extraction, and polymerase chain reaction (PCR) amplification. The molecular characterization was conducted using the TBR primer set, which targeted repeated satellite DNA regions and the cytochrome oxidase II gene of T. evansi. About 22.08% (53/240) of overall samples were positive for trypanosomiasis, with prevalence rates being 23.75% (19/80), 21.25% (17/80), and 21.75% (17/80) for districts Muzaffargarh, Lodhran, and Bahawalpur, respectively. 5.83% (14/240) of samples tested for T. evansi using PCR were positive in the districts of Muzaffargarh 7.50% (6/80), Lodhran 5.00% (4/80), and Bahawalpur 5.00% (4/80). Among the animals tested, camels had the highest positivity rate. The microscopic examination confirmed infection rates of 45.83% (22/48) for camels, 18.75% (9/48) for cattle, 8.33% (4/48) for buffaloes, 18.75% (9/48) for goats, and 18.75% (9/48) for sheep (p < 0.001). PCR results did not reveal substantial differences (p < 0.05) in prevalence: camels 12.50% (6/48), cattle 6.25% (3/48), buffaloes 0% (0/48), goats 8.33% (4/48), sheep 2.08% (1/48); while distinct disparities were detected district-wise: Muzaffargarh 23.75% (19/80), Lodhran 21.25% (17/80), and Bahawalpur 21.25% (17/80). The PCR results for these districts were insignificantly different: 7.50% (6/80), 5% (4/80), and 5% (4/80). The microscopic infection rate in camels from Bahawalpur was 56.30% (9/16). The microscopic analysis in Buffaloes reported a 6.30% (1/16) infection rate, but PCR results indicated no infections (0%) in any district. A significant difference (p < 0.001) in identifying Trypanosoma species was found between positively and negatively tested animals in both microscopic and PCR methods. This study emphasizes the necessity of regularly using PCR-based screening for its superior sensitivity and specificity over traditional microscopy. The varying occurrence of trypanosomiasis among districts reflects the intricate nature of this diseases epidemiology in the region. Reducing economic losses from trypanosomiasis in Southern Punjab, Pakistan, requires targeted interventions, such as vector control measures and farmer education.

Tick-borne piroplasms and trypanosomes incidentally detected in eastern grey kangaroos (Macropus giganteus) during a mortality and morbidity event in southern New South Wales, Australia

Tick-borne haemoparasites, including piroplasms and trypanosomes, are almost ubiquitous in Australian wildlife, with some associated with health impacts to individual animals and declining wildlife populations. An array of ecologically distinct piroplasm and trypanosome species occur throughout Australia although many of these species and their sylvatic ecologies are poorly characterised. Between May 2022 and October 2023, an anecdotally reported localised eastern grey kangaroo (Macropus giganteus) morbidity/mortality event occurred in coastal southern New South Wales, Australia, characterised by animals presenting with blindness, emaciation, lethargy, ataxia, and astasia. Here we used molecular techniques to identify tick-borne piroplasms (Babesia and Theileria) and trypanosomes in affected animals. Blood (n = 89) and liver (n = 19) samples were collected after the humane euthanasia of wild animals due to welfare concerns, and brief notes on the animal's health were recorded. In total, 20 (22.5%) animals were infected with tick-borne haemoparasites, including a novel Theileria sp. nov. (14, 15.7%), Babesia macropus (2, 2.2%), Trypanosoma gilletti (5, 5.6%), and Trypanosoma vegrandis (1, 1.1%). Liver samples were also screened for Wallal and Warego viruses due to animals' blindness, but were negative. This is the first report of T. gilletti and T. vegrandis in eastern grey kangaroos, although they have been previously reported in high numbers in ticks which commonly parasites this host. The novel Theileria sp. was previously reported in questing Ixodes holocyclus and in ticks from an opportunistically collected eastern grey kangaroo and red-necked wallaby (Notamacropus rufogriseus). However, we show for the first time this Theileria sp. can occur widely in eastern grey kangaroos. Ultimately, this small study did not intend, and is not able to draw inference regarding the pathogenicity of these haemoparasites to eastern grey kangaroos and it is likely that other factors, such as chronic Phalaris grass toxicity, had a role in this localised mortality/morbidity event.

Trypanocide Use and Molecular Characterization of Trypanosomes Resistant to Diminazene Aceturate in Cattle in Northern Côte D'Ivoire.

The resistance of trypanosomes to the doses of trypanocide administered by farmers to their animals acts as a real brake on efforts to control to combat African trypanosomiasis. Thus, in-depth knowledge of the use of these different molecules and their resistance profiles will be necessary to establish an integrated strategy to combat African trypanosomiasis. To achieve these objectives, a participatory survey among farmers and a resistance diagnosis of trypanosome strains identified in three regions of northern Côte d'Ivoire (Bagoué, Poro and Tchologo) was carried out using the PCR-RFLP technique, followed by sequencing of genes of interest. This study made it possible to identify three molecules that are commonly used by 85% (63/74) of farmers. In descending order of use, we identified Isometamidium chloride (43%), Diminazene aceturate (28%) and Homidium bromide (14%). Three species of trypanosomes, Trypanosoma congolense, Trypanosoma. theileri and Trypanosoma vivax, were identified in farms, and only one strain had the adenosine transporter gene (Trypanosoma congolense), but this strain was sensitive to the Diminazene aceturate molecule. Comparison of the sequence of this trypanosome strain showed that it is different to the Kenyan strain diagnosed as resistant to the Diminazene aceturate molecule. This study shows that a variety of trypanocides are used by farmers, and that the resistance profile of the strains to the Diminazene aceturate molecule could not be observed. However, it is important to further investigate the other molecules encountered in Côte d'Ivoire.