Improved serodiagnosis of Trypanosoma vivax infections in cattle reveals high infection rates in the livestock regions of Argentina.

Bovine trypanosomosis, caused by Trypanosoma vivax, currently affects cattle and has a significant economic impact in sub-Saharan Africa and South America. The development of new diagnostic antigens is essential to improve and refine existing methods. Our study evaluated the efficacy of two recombinant antigens in detecting specific antibodies in cattle. These antigens are derivatives of an invariant surface glycoprotein (ISG) from T. vivax. A fraction of a previously described antigen (TvY486_0045500), designated TvISGAf, from an African strain was evaluated, and a new ISG antigen from an American isolate, TvISGAm, was identified. The two antigens were expressed as fusion proteins in Escherichia coli: TvISGAf was fused to the MBP-His-tag, and TvISGAm was obtained as a His-tag fused protein. An ELISA evaluation was conducted using these antigens on 149 positive and 63 negative bovine samples. The diagnostic performance was enhanced by the use of a combination of both antigens (referred to as TvISG-based ELISA), achieving a sensitivity of 89.6% and specificity of 93.8%. Following the validation of the TvISG-based ELISA, the seroprevalence of T. vivax infection in 892 field samples from cattle in the central region of Argentina was determined. The mean seroprevalence of T. vivax was 53%, with variation ranging from 21% to 69% among the six departments studied. These results support the use of the TvISG ELISA as a valuable serological tool for the detection and monitoring of T. vivax infection in cattle. Furthermore, we report for the first time the seroprevalence of T. vivax in Argentina, which highlights the widespread endemic nature of the disease in the region. In order to effectively manage the increasing spread of T. vivax in the vast livestock production areas of South America, it is essential to implement consistent surveillance programs and to adopt preventive strategies.

Hemopathogens in naturally infected bovine fetuses in Brazil.

The transplacental transmission of parasites and hemoparasites is crucial for understanding the epidemiology of diseases. This study aimed to assess the prevalence of hemopathogens in bovine fetuses at various gestational periods. Samples were obtained from a slaughterhouse in the state of Minas Gerais, Brazil, and a total of 236 fetuses were collected. DNA extracted from blood samples (145) and organ samples (a pool of brain and spleen) (236) underwent a nested PCR (nPCR) assay to detect Babesia spp., Theileria spp., Trypanosoma vivax, Anaplasma marginale, Anaplasma bovis, Anaplasma phagocytophilum, Ehrlichia minasensis, and hemotropic Mycoplasma spp. Additionally, serological analysis of 145 plasma samples was conducted using the indirect fluorescent antibody test-IFAT to detect IgG against Babesia bovis, Babesia bigemina, A. marginale, and Trypanosoma vivax. The observed prevalence of transplacental transmission was 19.3 %, 6.2 %, 42.7 % and 2.7 %, for A. marginale, B. bigemina, 'Candidatus M. haemobos', and Mycoplasma wenyonii, respectively. The prevalence of A. marginale by gestational trimester was 16 % (13/81) in the second trimester and 23 % (14/60) in the third trimester, with no positive samples in the first trimester. Regarding the species B. bovis and B. bigemina, all evaluated animals tested negative by nPCR, and no serological evidence for B. bovis was found by the IFAT. Babesia bigemina demonstrated an overall seroprevalence of 6.2 % (9/145), with 4.8 % (7/145) in the last trimester and 1.3 % (2/145) in the second trimester of pregnancy. In total, 42.7 % (62/145) of blood samples were positive for 'Candidatus M. haemobos', with 42 % (34/81) in the middle trimester, and 43 % (26/60) in the final trimester of pregnancy. Mycoplasma wenyonni was detected in 2.7 % (4/145) blood samples, all in coinfection with 'C. M. haemobos'. The prevalence by pregnancy trimester was 25 % (1/4) in the first trimester; 1.2 % (1/81) in the second trimester and 3.3 % (2/60) in the third trimester of pregnancy. Hemopathogen DNA was detected in fetus blood samples but not the brain or spleen samples. All the samples were negative for T. vivax, Theileria spp., Anaplasma spp. and Ehrlichia spp. Overall, in this study, approximately 70 % of fetuses were positive for one or more of the studied parasites. No significant associations were observed between pairs of pathogens, except 'C. M. haemobos' and A. marginale.

First identification of Trypanosoma vivax among camels (Camelus dromedarius) in Yazd, central Iran, jointly with Trypanosoma evansi.

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.

Molecular Diagnosis and Prevalence of Trypanosoma vivax (Trypanosomatida: Trypanosomatidae) in Buffaloes and Ectoparasites in the Brazilian Amazon Region.

Trypanosoma vivax Ziemann is a parasite that affects both wild and domestic ungulates and is transmitted mechanically via tabanids and other blood-sucking insects in the Americas. A total of 621 blood samples from water buffaloes (Bubalus bubalis (Linnaeus) (Artiodactyla: Bovidae), and 184 ectoparasite samples (Amblyomma cajennense (Fabricius) sensu stricto and Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae), and Haematopinus tuberculatus (Burmeister) (Phthiraptera: Haematopinidae)) were obtained from 60 farms in the State of Pará, Brazilian Amazon. Twelve buffalo blood samples (1.89%) and 11 ectoparasites (6%) were positive for T. vivax based on the cathepsin L-like gene. All sequences were 99% similar to T. vivax from northeastern Brazil (EU753788) in amplified PCR assays on each of the hosts tested.

First report and molecular identification of Trypanosoma (Duttonella) vivax outbreak in cattle population from Ecuador.

The bovine trypanosomosis is responsible for economic losses from tropical and subtropical areas of Africa and Latin America. This disease is characterized by fever, anaemia, loss of production and even death. Few studies have been carried out in Ecuador regarding Trypanosoma spp. presence but the species has not been determined in cattle and those have only determined the presence of genus, but not the species. The aim of this study was to identify and characterize the trypanosome species involved in the suspected bovine trypanosomosis outbreak reported in Convento Village in Manabí Province located in the coastal region of Ecuador. Twenty cattle from three farms were sampled. Three samples were positive for T. vivax, using an end-point polymerase chain reaction (PCR) to amplify a fragment of the cathepsin L-like cysteine protease (CatL-like) gene. A phylogenetic tree analysis of these three Ecuadorian isolates showed a close relationship with isolates from South America (Colombia, Brazil and Venezuela) and West Africa (Nigeria). This is the first report of T. vivax in Ecuadorian cattle.

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

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

Epidemiological and molecular identification of Trypanosoma vivax diagnosed in cattle during outbreaks in central Brazil.

Bovine trypanosomosis has been spreading in Brazil. In the present study, we evaluated the spatial distribution, prevalence and risk factors of this disease in the state of Goiás, Brazil, and performed both molecular and phylogenetical analyses of Trypanosoma vivax. A total of 4049 blood samples were collected from cattle for a period of 2 years. The parasitological diagnosis was performed using the Woo method and a questionnaire was administered to the farmers to document risk factors associated with the disease in the herd. Positive samples were DNA sequenced and compared to GenBank codes. The prevalence of T. vivax was 8.84%, occurring on 24 ranches only in dairy cattle and mainly in the central and southern portions of the state. The acquisition of new animals infected with T. vivax and the administration of exogenous oxytocin to cows using the same syringe and needle were the main associated factors (P ≤ 0.05). After an outbreak, milk production decreased by 39.62%. The presence of biting flies (tabanids, Haematobia irritans and Stomoxys calcitrans) was not a risk factor (P > 0.05) for the occurrence of T. vivax. The epidemiological data demonstrate the importance of restricting the practice of auctions as well as eliminating the use of exogenous oxytocin in animals during milking. The samples tested by polymerase chain reaction were positive for T. vivax and were genetically homologous with T. vivax found in different states of Brazil and west Africa based on the 18S rRNA gene.

High Trypanosoma vivax infection rates in water buffalo and cattle in the Brazilian Lower Amazon.

Highly sensitive and accurate molecular diagnostic methods have not yet been employed for livestock trypanosomosis in the Brazilian Lower Amazon although the first reports of Trypanosoma vivax and Trypanosoma evansi in Brazil were in water buffalo (Bubalus bubalis) in this region. The present study assessed trypanosomosis in buffalo and cattle raised in communal and seasonally flooding pastures in the state of Pará using the fluorescent fragment length barcoding (FFLB) method. T. evansi was not detected, but high infection rates of T. vivax and T. theileri were revealed by a simplified FFLB standardized in the present study that discriminates all trypanosome species infective to livestock in South America. T. vivax infection rates detected by TviCATL-PCR were 24.6% for cattle (n = 61) and 28.1% for buffalo (n = 89). Using the FFLB method, overall T. vivax infection rates increased to 59.6% and 44.3% for buffalo and cattle, respectively. Furthermore, the predominance of a single microsatellite-based genotype of T. vivax was reinforced in the Lower Amazon. Relevant T. vivax infection rates detected in clinically healthy buffalo and cattle through the sampled years (2008-2017) highlight the need for systematic studies to demonstrate the endemic steady state of T. vivax in this region. Our findings provide baseline information for livestock management, including control of T. vivax dispersal, and the introduction of naïve animals. The growing international trade of live livestock from this very important livestock breeding region represents a serious risk for T. vivax spreading outside Amazonia and Brazil.

Possible influence of Plasmodium/Trypanosoma co-infections on the vectorial capacity of Anopheles mosquitoes.

OBJECTIVE: In tropical Africa, trypanosomiasis is present in endemic areas with many other diseases including malaria. Because malaria vectors become more anthropo-zoophilic under the current insecticide pressure, they may be exposed to trypanosome parasites. By collecting mosquitoes in six study sites with distinct malaria infection prevalence and blood sample from cattle, we tried to assess the influence of malaria-trypanosomiasis co-endemicity on the vectorial capacity of Anopheles. RESULTS: Overall, all animal infections were due to Trypanosoma vivax (infection rates from 2.6 to 10.5%) in villages where the lowest Plasmodium prevalence were observed at the beginning of the study. An. gambiae s.l. displayed trophic preferences for human-animal hosts. Over 84 mosquitoes, only one was infected by Plasmodium falciparum (infection rate: 4.5%) in a site that displayed the highest prevalence at the beginning of the study. Thus, Anopheles could be exposed to Trypanosoma when they feed on infected animals. No Plasmodium infection was observed in the Trypanosoma-infected animals sites. This can be due to an interaction between both parasites as observed in mice and highlights the need of further studies considering Trypanosoma/Plasmodium mixed infections to better characterize the role of these infections in the dynamic of malaria transmission and the mechanisms involved.

Trypanosome infection rate in Glossina tachinoides: infested rivers of Limmu Kosa District Jimma Zone, Western Ethiopia.

OBJECTIVE: Trypanosomosis is a disease of domestic animals and humans resulting from infection with parasitaemic protozoa of the genus Trypanosoma transmitted primarily by tsetse flies. A cross-sectional study was conducted from January-March 2018, to estimate the infection rate of trypanosome in Glossina tachinoides, their distribution, magnitude and involved trypanosome species in Limmu Kosa District of Jimma zone. RESULTS: Study methodology involved entomological survey using monoconical traps to study the magnitude of Fly density Flay/Trap/Day (FTD) and tsetse fly dissection to estimate infection rate of trypanosome in vector flies. The study result indicated that there was only one species of Tsetse fly Glossina tachinoides detected with FTD = 4.45. From the total of (n = 284) dissected Glossina tachinoides flies only (n = 5) positive for Trypanosome resulting in 1.76% Infection Rate. Peak trypanosome infections were observed in female tsetse 2.04%, n = 4 and 1.14%, n = 1 in males. Furthermore, 1.06% of Glossina tachinoides were infected by Trypanosome vivax and the remaining 0.70% was Trypanosome congolense. Finally, the study concluded with the recommendation of control and suppression of the vector and parasite was mandatory due to Pathogenic Animal Trypanosomosis.

Prevalence of bovine trypanosomosis and vector density in a dry season in Gamo-Gofa and Dawuro Zones, Southern Ethiopia.

Trypanosomosis remains one of the biggest constraints of livestock productivity in sub-Saharan Africa. It is of particular concern in Ethiopia where crop production is largely dependent on animal traction power. This study was conducted between November 2015 and March 2016 to estimate the prevalence of bovine trypanosomosis and vector density in Gamo-Gofa and Dawuro Zones located in Southern Ethiopia. For the entomological survey, a total of 305 NGU traps were deployed for three consecutive days at different positions in the two study areas. For parasitological study, blood samples were collected from 2402 cattle and examined for the presence of trypanosomes by the buffy coat technique (BCT). Blood samples that were positive in the BCT were further tested with Giemsa-stained thin smears for identification of Trypanosoma species. In the entomological survey, a total of 4113 flies were captured of which 1605 (39%) were tsetse flies while 2508 (61%) were other biting flies of the genus Stomoxys and Tabanus. Glossina pallidipes was the only tsetse fly identified in the study areas. The overall mean apparent density of G. pallidipes was 1.8 flies per trap per day (FTD). The overall prevalence of trypanosomosis in the study areas was 5.1% (95% CI: 4.2-6.0); however, it was significantly (p < 0.001) higher in Dawuro Zone (10%) than Gamo-Gofa (1.9%). Prevalence was also noted to be significantly (p < 0.001) higher in cattle with black coat colour and those with poor body condition. Trypanosomosis was caused mainly by Trypanosoma congolense (61.8%) and to a lesser extent by T. vivax (28.5%). Mixed infection by the two spp. was seen in 9.8% of the total positive animals. Evaluation of the mean packed cell volume (PCV) of the study animals with infection status revealed a significantly (p < 0.001) lower mean PCV in parasitaemic animals (21.5 ±â€¯0.46SE) than aparasitaemic ones (24.3 ±â€¯0.11SE). Generally, the prevalence noted in the current study is moderate. However the observation of such level of infection in a dry season suggests that the disease is still an important constraint of cattle production in the study areas. Thus, we recommend that theexisting parasite and vector control interventions need to be strengthened with special emphasis to Dawuro Zone where the prevalence was significantly higher. As the current sampling was done only once and in a dry season only, further longitudinal studies including all the seasons of the year need to be considered in the future.

Genetic diversity of trypanosome species in tsetse flies (Glossina spp.) in Nigeria.

BACKGROUND: Trypanosomes cause disease in humans and livestock in sub-Saharan Africa and rely on tsetse flies as their main insect vector. Nigeria is the most populous country in Africa; however, only limited information about the occurrence and diversity of trypanosomes circulating in the country is available. METHODS: Tsetse flies were collected from five different locations in or adjacent to protected areas, i.e. national parks and game reserves, in Nigeria. Proboscis and gut samples were analysed for trypanosome DNA by molecular amplification of the internal transcribed spacer 1 (ITS1) region and part of the trypanosome specific glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) gene. RESULTS: The most abundant Trypanosoma species found in the tsetse gut was T. grayi, a trypanosome infecting crocodiles. It was ubiquitously distributed throughout the country, accounting for over 90% of all cases involving trypanosomes. Trypanosoma congolense was detected in gut samples from all locations except Cross River National Park, but not in the proboscis, while T. brucei (sensu lato) was not detected at all. In proboscis samples, T. vivax was the most prominent. The sequence diversity of gGAPDH suggests that T. vivax and T. grayi represent genetically diverse species clusters. This implies that they are highly dynamic populations. CONCLUSIONS: The prevalence of animal pathogenic trypanosomes throughout Nigeria emphasises the role of protected areas as reservoirs for livestock trypanosomes. The genetic diversity observed within T. vivax and T. grayi populations might be an indication for changing pathogenicity or host range and the origin and consequences of this diversity has to be further investigated.

A national atlas of tsetse and African animal trypanosomosis in Mali.

BACKGROUND: Tsetse-transmitted trypanosomosis is a deadly, neglected tropical disease and a major challenge for mixed crop-livestock agriculture in sub-Saharan Africa. It is caused by several species of the genus Trypanosoma. Information on the occurrence of tsetse flies and African animal trypanosomosis (AAT) is available for different areas of Mali. However, these data have never been harmonized and centralized, which prevents the development of comprehensive epidemiological maps and constrains an evidence-based planning of control actions. To address this challenge, we created a dynamic geo-spatial database of tsetse and AAT distribution in Mali. METHODS: A digital repository containing epidemiological data collected between 2000 and 2018 was assembled. In addition to scientific publications, the repository includes field datasheets, technical reports and other grey literature. The data were verified, harmonized, georeferenced and integrated into a single spatially-explicit database. RESULTS: For the tsetse component, approximately 19,000 trapping records, corresponding to 6000 distinct trapping locations and 38,000 flies were included in the database. Glossina palpalis gambiensis was the most widespread and abundant species, and it was found in the southern, southern-central and western parts of the country. Glossina tachinoides was only found in the South. Only a few specimens of Glossina morsitans submorsitans were detected. For the AAT component, approximately 1000 survey records were included, corresponding to 450 distinct survey sites and 37,000 tested bovines. AAT was found in all surveyed regions, although data for the tsetse-free North and North-East are lacking. Trypanosoma vivax and Trypanosoma congolense were the dominant species, while Trypanosoma brucei infections were much less numerous. CONCLUSIONS: The atlas of tsetse and AAT in Mali provides a synoptic view of the vector and disease situation at the national level. Still, major geographical gaps affect the North, the North-East and the West, and there is also a severe lack of data over the past five years. Trypanosomosis remains a major animal health problem in Mali. However, despite its prevalence and distribution, monitoring and control activities are presently very limited. Efforts should be made to strengthen the progressive control of AAT in Mali, and the atlas provides a new tool to identify priority areas for intervention.

Cardiac involvement in trypanosomiasis in sheep experimentally infected by Trypanosoma vivax (Ziemman, 1905).

The objective of the present study was to evaluate the clinical signs, electrocardiographic signs and evolution of histopathological lesions in the heart of sheep experimentally infected by Trypanosoma vivax during the acute and chronic phases of infection as well as to investigate the presence of parasitic DNA in the heart using polymerase chain reaction (PCR). Twenty-two male sheep were divided into the following four groups: G1, which consisted of six sheep infected by T. vivax that were evaluated until 20 days post-infection (dpi; acute phase); G2, which consisted of six sheep infected by T. vivax that were evaluated until 90 dpi (chronic phase); and G3 and G4 groups, which each consisted of five uninfected sheep. At the end of the experimental period, electrocardiographic evaluations and necroscopic examinations were performed. Fragments of the heart were collected and stained by Hematoxylin-Eosin and Masson's trichrome, and the fragments were also evaluated by PCR for T. vivax. G2 animals presented clinical signs suggestive of heart failure and electrocardiogram alterations characterized by prolonged P, T and QRS complex durations as well as by a cardiac electrical axis shift to the left and increased heart rate. In these animals, mononuclear multifocal myocarditis and interstitial fibrosis were also observed. PCR revealed positivity for T. vivax in two G1 animals and in all G2 animals. Thus, these findings suggested that T. vivax is responsible for the occurrence of cardiac lesions, which are related to heart failure, electrocardiographic alterations and mortality of the infected animals.

Epizootic Infection by Trypanosoma vivax in Cattle from the State of Minas Gerais, Brazil.

Trypanosomiasis is caused by a pathogenic protozoan of the genus Trypanosoma, being Trypanosoma vivax the most important agent for cattle. The aim of the present study was to demonstrate the expansion of T. vivax infection in different mesoregions of Minas Gerais, Brazil, and describe the clinicopathological findings of trypanosomiasis in cattle. The diagnosis was based on visualization of the parasite in blood smears and DNA detection of T. vivax in the blood of live cows and tissues of necropsied animals by the polymerase chain reaction (PCR). Thirty suspected herds were tested, of which 11 were positive for T. vivax. The most frequent clinical signs were anemia, apathy, drop in milk production, weight loss, reproductive disorders, and nervous signs. Concomitant diseases, such as malignant edema, pneumonia and increased cases of mastitis were associated with T. vivax infection. Three cows were necropsied and the most significant findings were low body condition score, pale mucous and spleen with white pulp hyperplasia. The results demonstrated the expansion of T. vivax infection in Minas Gerais, that PCR-associated blood smears are promising for diagnosis, and that other diseases often occur concomitantly to T. vivax infection in regions with trypanosomiasis in cattle.

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

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

Remarkable richness of trypanosomes in tsetse flies (Glossina morsitans morsitans and Glossina pallidipes) from the Gorongosa National Park and Niassa National Reserve of Mozambique revealed by fluorescent fragment length barcoding (FFLB).

Trypanosomes of African wild ungulates transmitted by tsetse flies can cause human and livestock diseases. However, trypanosome diversity in wild tsetse flies remains greatly underestimated. We employed FFLB (fluorescent fragment length barcoding) for surveys of trypanosomes in tsetse flies (3086) from the Gorongosa National Park (GNP) and Niassa National Reserve (NNR) in Mozambique (MZ), identified as Glossina morsitans morsitans (GNP/NNR=77.6%/90.5%) and Glossina pallidipes (22.4%/9.5%). Trypanosomes were microscopically detected in 8.3% of tsetse guts. FFLB of gut samples revealed (GNP/NNR): Trypanosoma congolense of Savannah (27%/63%), Kilifi (16.7%/29.7%) and Forest (1.0%/0.3%) genetic groups; T. simiae Tsavo (36.5%/6.1%); T. simiae (22.2%/17.7%); T. godfreyi (18.2%/7.0%); subgenus Trypanozoon (20.2%/25.7%); T. vivax/T. vivax-like (1.5%/5.2%); T. suis/T. suis-like (9.4%/11.9%). Tsetse proboscises exhibited similar species composition, but most prevalent species were (GNP/NNR): T. simiae (21.9%/28%), T. b. brucei (19.2%/31.7%), and T. vivax/T. vivax-like (19.2%/28.6%). Flies harboring mixtures of trypanosomes were common (~ 64%), and combinations of more than four trypanosomes were especially abundant in the pristine NNR. The non-pathogenic T. theileri was found in 2.5% while FFLB profiles of unknown species were detected in 19% of flies examined. This is the first report on molecular diversity of tsetse flies and their trypanosomes in MZ; all trypanosomes pathogenic for ungulates were detected, but no human pathogens were detected. Overall, two species of tsetse flies harbor 12 species/genotypes of trypanosomes. This notable species richness was likely uncovered because flies were captured in wildlife reserves and surveyed using the method of FFLB able to identify, with high sensitivity and accuracy, known and novel trypanosomes. Our findings importantly improve the knowledge on trypanosome diversity in tsetse flies, revealed the greatest species richness so far reported in tsetse fly of any African country, and indicate the existence of a hidden trypanosome diversity to be discovered in African wildlife protected areas.

Occurrence of trypanosome infections in cattle in relation to season, livestock movement and management practices of Maasai pastoralists in Northern Tanzania.

African animal trypanosomosis (AAT) is a parasitic disease considered to be one of the greatest constraints to cattle production in Tanzania. There is insufficient information on seasonal occurrence of AAT and management practices in Monduli District of the Maasai Steppe ecosystem to guide and prioritize AAT control programs. A cross-sectional survey was undertaken in 10 randomly selected villages of Monduli District. Information on seasonal animal movements, including wildlife interaction, and AAT management practices was gathered using a standardized questionnaire with 130 pastoralists. Blood samples were collected from a total of 960 cattle, in wet and dry seasons. An entomological survey was also undertaken in the dry season. Polymerase chain reaction targeting the internal transcribed spacer 1 (ITS1) was used for parasite identification in cattle blood and in tsetse flies. The overall apparent prevalence of AAT in cattle was 5.8% (95% CI of 4.1-8.3) and 4.2% (95% CI of 2.7-6.3) during wet and dry reasons, respectively. Trypanosoma vivax was the most common species identified in cattle. All tsetse flies (n = 426) collected were trapped in Esilalei village which is in close proximity to Lake Manyara National Park. Tsetse fly infection status was determined to be 7.0%; (CI 95% of 4.9-9.8%) with nearly 50% of infections due to T. congolense. All 130 cattle owners reported that they could easily recognize AAT and the majority (75%) identified the most prominent clinical signs. Nearly all owners (98.5%) identified that tsetse flies were responsible for AAT transmission. All cattle owners (100%) reported the use of trypanocides for AAT treatment, while 2.3% reported to also use herbal medicine. The trypanocides, Novidium® and Berenil®, were the most frequently reported commercial drugs and were used by 42% of cattle owners. Vector control by hand spraying was reported by the majority (90.8%) of cattle owners, while dipping and deployment of insecticide-impregnated targets were reported by few cattle owners (16.2% and 5.4%, respectively). The majority of cattle owners (83.1%) reported to move cattle away from home villages during the dry season with many migrating to areas in close proximity to wildlife parks. This study confirms the presence of circulating pathogenic trypanosomes in tsetse flies which continue to pose a threat to Maasai cattle. The seasonal movement of cattle during the dry season was associated with more clinical cases of cattle trypanosomosis. This study demonstrates the need to strengthen surveillance and control strategies for AAT.

Detection and molecular characterization of Trypanosoma (Duttonella) vivax in dairy cattle in the state of Sergipe, northeastern Brazil / Detecção e caracterização molecular de Trypanosoma (Duttonella) vivax em gado leiteiro no estado de Sergipe, no Nordeste do Brasil

Abstract Trypanosoma (Duttonella) vivax is an important cause of economic losses among feedlot cattle. These losses are related to the morbidity, mortality, reproductive issues and decreased production. It is known that the clinical signs observed in infections by this protozoon are similar to other hemoparasitosis, which difficult the diagnosis. Therefore, the aim of this study was to detect and molecularly characterize an outbreak of trypanosomiasis caused by T. (D.) vivax in dairy cattle in the municipality of São Miguel Aleixo, state of Sergipe, Brazil. Blood samples from cattle (n = 15) presenting clinical signs compatible with trypanosomiasis were collected and parasitological and molecular evaluated. Among the samples analyzed, 34% (5/15) were positive from blood smears, 60% (9/15) from the buffy coat method and 80% (12/15) from the molecular method. The DNA sequence obtained (659 bp) showed 99% similarity to T. (D.) vivax sequences that are available in the GenBank database. The presence of this protozoon in cattle herds is a problem for producers. Diagnosing trypanosomiasis is problematic because its evolution is similar to that of other parasitic blood diseases. In addition, this is the first report of infection by T. (D.) vivax in cattle in the state of Sergipe, northeastern Brazil.

Resumo Trypanosoma (Duttonella) vivax é responsável por consideráveis perdas econômicas na bovinocultura. Estas perdas estão relacionados à morbidade, mortalidade, problemas reprodutivos e declínio na produção. Sabe-se que os sinais clínicos apresentados em infecções por este protozoário se assemelha a outras hemoparasitoses, dificultando muitas vezes o diagnóstico. Portanto, objetivou-se com este estudo detectar a ocorrência de T. (D.) vivax em bovinos leiteiros no município de São Miguel Aleixo, Estado de Sergipe, Brasil. Para tanto, amostras de sangue (n = 15) foram coletadas e avaliadas através de métodos parasitológicos e moleculares. Do total das amostras analisadas, 34% (5/15) foram positivas no esfregaço sanguíneo, 60% (9/15) pelo método do Buffy Coat, enquanto na biologia molecular 80% (12/15) amplificaram um fragmento de DNA (659 pb) compatível com T. (D.) vivax (GenBank). Em conclusão a presença de T. (D.) vivax nos rebanhos bovinos caracteriza-se como um problema para os pecuaristas, como também para o diagnóstico, uma vez que essa tripanossomíase apresenta evolução semelhante a outras hemoparasitoses. Ademais, este é o primeiro relato de infecção por T. (D.) vivax em bovinos do estado de Sergipe, nordeste do Brasil.

Detection and molecular characterization of Trypanosoma (Duttonella) vivax in dairy cattle in the state of Sergipe, northeastern Brazil.

Trypanosoma (Duttonella) vivax is an important cause of economic losses among feedlot cattle. These losses are related to the morbidity, mortality, reproductive issues and decreased production. It is known that the clinical signs observed in infections by this protozoon are similar to other hemoparasitosis, which difficult the diagnosis. Therefore, the aim of this study was to detect and molecularly characterize an outbreak of trypanosomiasis caused by T. (D.) vivax in dairy cattle in the municipality of São Miguel Aleixo, state of Sergipe, Brazil. Blood samples from cattle (n = 15) presenting clinical signs compatible with trypanosomiasis were collected and parasitological and molecular evaluated. Among the samples analyzed, 34% (5/15) were positive from blood smears, 60% (9/15) from the buffy coat method and 80% (12/15) from the molecular method. The DNA sequence obtained (659 bp) showed 99% similarity to T. (D.) vivax sequences that are available in the GenBank database. The presence of this protozoon in cattle herds is a problem for producers. Diagnosing trypanosomiasis is problematic because its evolution is similar to that of other parasitic blood diseases. In addition, this is the first report of infection by T. (D.) vivax in cattle in the state of Sergipe, northeastern Brazil.