Entomological study on vectorial density, temporal variation of tsetse fly and other biting flies in intervention and non-intervention areas of South Omo Zone, Ethiopia.

A repeated cross-sectional entomological survey was conducted to estimate Glossina (tsetse) and other biting flies density, their seasonal variation and associated risk factors in intervention and non- intervention areas of South Omo Zone, Southwest Ethiopia from January 2019-November 2019. In both dry and wet seasons, a total of 96 NGU traps (64 traps in tsetse intervention districts and 32 traps in tsetse non- intervention districts) were deployed at an interval of about 100-200 m in purposively selected and suspected tsetse habitats. Thus, Glossina pallidipes was found to be the only cyclical vector along with mechanical vectors of Tabanus, Stomoxys and Haematopota. In tsetse intervention areas, G. pallidipes apparent density of 2.64 F/T/D and 0.42 F/T/D was recorded in dry and wet season respectively. Mechanical vectors (dry; wet) of Tabanus (205; 155), Stomoxys (34; 54) and Haematopota (50; 33) were also recorded in tsetse intervened areas. Whereas, in non- intervention areas, apparent density of G. pallidipes was 2.03 F/T/D and 0.56 F/T/D, respectively in dry and wet season. Similarly, Tabanus (22; 56), Stomoxys (10; 8) and Haematopota (5; 7) respectively in dry and wet (dry; wet) season were recorded in tsetse non- intervention areas. According to Negative Binomial Regression (NBR), season was the only variable significantly affecting (P < 0.05) the Glossina count in the current study area. Accordingly, the incidence G. pallidipes during wet season was decreased by the factor of 0.21 (CI; 0.097-0.47) when compared to its incidence in dry season by holding other variables constant. In conclusion, cyclical vectors were playing vital role in transmission of trypanosomosis in South Omo Zone along with numerous mechanical vectors even though there have been vector intervention activities in the areas. Therefore, strong, sustainable, environmentally friend and community participating vector control strategies should be followed to tackle the vector distribution in the area.

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

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

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

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

Parasitological investigation of bovine Trypanosomosis, vector distribution and tsetse flies infection rate study, Dabo Hana District, Buno Bedelle Zone, Southwest Ethiopia.

A cross-sectional study was conducted to explore the prevalence of Trypanosome infections in cattle and within the tsetse flies from December 2020 to May 2021 in Dabo Hana district, Buno Bedelle Zone, Southwest Ethiopia. A total of 415 blood samples were examined utilizing Buffy coat and Giemsa-stained thin blood smear techniques. Vector distribution and tsetse fly infection rate were studied by deploying 60 traps in four purposively chosen villages of the district. The prevalence of Trypanosomes was 10.6% and 6.5% in cattle and in tsetse flies, respectively. Trypanosoma congolense (59.1%) in cattle and T. vivax (62.5%) in tsetse flies, were the foremost common species distinguished in the area. A significant difference (P ≤ 0.05) was observed in the prevalence of bovine Trypanosomosis between body condition scores of cattle. However, differences were not significant between coat color, sex, and age categories (P > 0.05). The mean PCV values of Trypanosome-infected cattle (22.6 ± 0.6) were significantly (P < 0.05) lower than those of non-infected cattle (25.6 ± 0.3). Out of 1441 flies caught, 1242 (86.2%) were Glossina, 113 (7.84%) were Stomoxys, and 86 (5.97%) were Tabanus. Of 1242 Glossina, 85% were G. tachinoides and the remaining 15% were G. m. sub-morsitans. This finding revealed that, three Trypanosoma species are circulating in cattle as well as in tsetse flies. It is recommended that, sustainable and integrated tsetse and Trypanosomosis control practices should be implemented to foster live stock health and agricultural development in the district. Other sensitive methods should be employed to determine the true picture of infection in the area.

Prevalence of bovine trypanosomiasis in Côte d'Ivoire: Systematic review and meta-analysis.

Bovine trypanosomiasis is a parasitic disease caused by protozoans of the genus Trypanosoma. The disease cause economic losses in livestock production. In order to determine the status of research on this disease in Côte d'Ivoire, we used the systematic review method and meta-analysis. Three electronics databases, namely Google Scholar, PubMed and CrossRef were used to search for publications on trypanosomiasis prevalence that met our inclusion criteria. Twenty five articles were identified, 11 of which met the inclusion criteria. Bovine trypanosomiasis prevalence of 2.99% (95% confidence interval [CI]: 2.96% - 3.01%) to 25.28% (95% CI: 25.17% - 25.38%) were recorded between 1960 and 2021. The analyses showed that the most infected regions were the Bagoue 11.26% (95% CI: 11.25% - 11.27%), Bounkani 14.94% (95% CI: 14.93% - 14.95%), Gbeke 10.34% (95% CI: 10.33% - 10.35%), Marahoue 13.79% (95% CI: 13.78% - 13.80%), Poro 8.50% (95% CI: 8.49% - 8.51%), and Tchologo 11.83% (95% CI: 11.82% - 11.84%).The most sensitive diagnostic method used was the polymerase chain reaction (PCR). The species of trypanosomes diagnosed were Typanosoma vivax 4.99% (95% CI: 4.97% - 5.01%), T. congolense 1.51% (95% CI: 1.49% - 1.52%), and T. brucei 0.61% (95% CI: 0.59% - 0.62%). Despite some variation, the prevalence of bovine trypanosomiasis in Côte d'Ivoire caused mainly by T. vivax has increased in the years between 1977 and 2017. Efforts to control tsetse and other mechanical vectors should also be put in place to minimize its transmission.Contribution: The authors studied the prevalence of bovine trypanosomiasis using the systematic review method and MA in order to determine the status of research on this disease in Côte d'Ivoire.

Methods Applied to the Diagnosis of Cattle Trypanosoma vivax Infection: An Overview of the Current State of the Art.

Bovine trypanosomiasis caused by Trypanosoma vivax is a relevant disease in domestic ungulates in Latin America, causing different types of livestock losses, particularly in African and South American countries, leading to loss of millions of dollars/year related to dairy and meat production. In addition, T. vivax trypanosomiasis requires intensive veterinary care. While vector control is a feasible measure to manage disease spreading, the search for accurate diagnostic tools still represents a gap in routine veterinary practices and a challenge for the scientific community. The parasite is mechanically transmitted by fomites or by the saliva of haematophagous flies, such as Stomoxys sp. and Tabanus sp., infecting cattle as well as a number of animal hosts. The main symptoms of T. vivax bovine trypanosomiasis are apathy, fever, restricted growth, miscarriage, progressive weakness, neurological signs, pale mucous, loss of appetite, lethargy, and substantial weight loss. In most cases, the presence of animals with subclinical infections, nonspecific symptoms and without apparent parasitaemia presents a challenge when making a diagnosis, which requires accurate methods. Herein, we review state of the art concerning current methods available for the diagnosis of T. vivax bovine trypanosomiasis, focusing on clinical, parasitological, immunological and molecular approaches, highlighting the main features of each method, including "pros and cons". Overall, combining several diagnostic techniques is a better choice since it leads to fewer false negative results and contributes to better disease control.

The economic cost of bovine trypanosomosis in pastoral and ago pastoral communities surrounding Murchision Falls National park, Buliisa district, Uganda.

BACKGROUND: Animal diseases that are endemic like tsetse transmitted trypanosomosis cause the continuous expenditure of financial resources of livestock farmers and loss of productivity of livestock. Estimating the cost of controlling animal trypanosomosis can provide evidence for priority setting and targeting cost-effective control strategies. METHODOLOGY: A cross-sectional survey to estimate the economic cost of bovine trypanosomosis was conducted in cattle-keeping communities living around Murchision falls National Park, in Buliisa district Uganda. Data was collected on herd structure, the cost of treatment and control, prevalence of morbidity and mortality rates due to trypanosomosis, and salvage sales losses in cattle herds in the last year. RESULTS: In this study, 55.4% (n = 87) of the households reported their cattle had been affected by trypanosomosis during the previous last year. There was a high economic cost of trypanosomosis (USD 653) per household in cattle-keeping communities in Buliisa district of which 83% and 9% were due to mortality and milk loss respectively/ High mortality loss was due to low investment in treatment. The study showed that prophylactic treatment 3 times a year of the whole herd of cattle using Samorin ® (Isometamidium chloride) at a cost of USD 110 could drastically reduce cattle mortality loss due to trypanosomosis due to trypanosomosis with a return on investment of USD 540 annually per herd. This could be coupled with strategic restricted insecticide spraying of cattle with deltamethrin products. CONCLUSION: The results show a high economic cost of trypanosomosis in cattle-keeping communities in Buliisa district, with cattle mortality contributing the largest proportion of the economic cost. The high mortality loss was due to low investment in treatment of sick cattle.

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

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

A cerumenolomic approach to bovine trypanosomosis diagnosis.

INTRODUCTION: Trypanosomiasis caused by Trypanosoma vivax (T. vivax, subgenus Duttonella) is a burden disease in bovines that induces losses of billions of dollars in livestock activity worldwide. To control the disease, the first step is identifying the infected animals at early stages. However, convention tools for animal infection detection by T. vivax present some challenges, facilitating the spread of the pathogenesis. OBJECTIVES: This work aims to develop a new procedure to identify infected bovines by T. vivax using cerumen (earwax) in a volatilomic approach, here named cerumenolomic, which is performed in an easy, quick, accurate, and non-invasive manner. METHODS: Seventy-eight earwax samples from Brazilian Curraleiro Pé-Duro calves were collected in a longitudinal study protocol during health and inoculated stages. The samples were analyzed using Headspace/Gas Chromatography-Mass Spectrometry followed by multivariate analysis approaches. RESULTS: The cerumen analyses lead to the identification of a broad spectrum of volatile organic metabolites (VOMs), of which 20 VOMs can discriminate between healthy and infected calves (AUC = 0.991, sensitivity = 0.967, specificity = 1.000). Furthermore, 13 VOMs can indicate a pattern of discrimination between the acute and chronic phases of the T. vivax infection in the animals (AUC = 0.989, sensitivity = 0.944, specificity = 1.000). CONCLUSION: The cerumen volatile metabolites present alterations in their occurrence during the T.vivax infection, which may lead to identifying the infection in the first weeks of inoculation and discriminating between the acute and chronic phases of the illness. These results may be a breakthrough to avoid the T. vivax outbreak and provide a faster clinical approach to the animal.

How many cattle can be infected by Trypanosoma vivax by reusing the same needle and syringe, and what is the viability time of this protozoan in injectable veterinary products?

It was investigated how many cattle become infected with Trypanosoma vivax by subcutaneous (SC), intramuscular (IM) and intravenous (IV) routes, using the same syringe and needle from an animal with acute T. vivax infection. Besides, the T. vivax viability in 109 injectable veterinary drugs (antibiotics, antiparasitics, reproductive hormones, vitamin complex and derivatives, vaccines, anaesthetics, anti-inflammatory/antipyretics, antitoxics). In the field assay, four groups were performed: T01, T02 and T03 animals that received saline solution with the same syringe and needle contaminated with T. vivax via SC, IM and IV routes, respectively, and T04 control animals that received only saline solution with the same syringe and needle IV. In the laboratory, drugs had their pH measured and T. vivax viability verified. The number of cattle infected with T. vivax via SC (3/20) was lower (P ≤ 0.05) compared to via IM (9/20), which was lower (P ≤ 0.05) compared to IV (15/20). The solution pH did not influence T. vivax viability. In 44% (48/109) of the products, T. vivax remained viable regardless of time, stooding out that in 100% of oxytocins the protozoan was verified, at some evaluation times. The mean of T. vivax quantified in foot-and-mouth and brucellosis vaccines and in doramectin-based products were higher (P ≤ 0.05) than found in blood + saline solution.

Evaluation of techniques for diagnosis of Trypanosoma vivax infections in naturally infected cattle in the Zona da Mata Mineira.

This study aimed to evaluate diagnostic techniques for trypanosomiasis, caused by Trypanosoma vivax, in naturally infected cattle in Minas Gerais, Zona da Mata. The deaths of six lactating cows with similar clinical conditions-characterized by hyporexia, hypogalactia, and recumbency-had been reported from one property. Initially, two animals were examined and diagnosed with trypanosomiasis through identification of the protozoan in a blood smear. After the initial diagnosis, all lactating cows (n=37) on the property were examined, and blood samples were collected for tests including whole blood smear, buffy coat smear, Woo's technique, enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction (PCR). Woo's test, buffy coat smears, and whole blood smears indicated that 4/37 (10.81%) animals were positive for trypanosomiasis, whereas ELISA and PCR indicated that 33/37 (89.19%) and 27/37 (72.97%) animals, respectively, were positive. The agreement obtained between parasitological techniques was classified as high, while between ELISA and PCR, no agreement. In conclusion, parasitological techniques have a low capacity to identify infected animals in the chronic stage of T. vivax infection. Therefore, techniques such as PCR and/or ELISA should be used to minimize the occurrence of false negatives.

Serum prevalence of anti-Trypanosoma vivax antibodies in cattle in Tapira-MG, Brazil / Soroprevalência de anticorpos anti-Trypanosoma vivaxem bovinos em Tapira-MG, Brasil

Trypanosoma vivax is considered the most important pathogenic Trypanosoma for cattle and causes great damage to the dairy and beef cattle industries. This study aimed to evaluate the prevalence of anti-T. vivax antibodies in dairy cattle from the municipality of Tapira, located in the Alto Paranaíba region, Minas Gerais, Brazil. The 74 blood serum samples from dairy cattle were analyzed using an indirect immunofluorescence reaction. The seroprevalence was 82.4 % (61/74), and the highest incidence observed can be correlated with the transit of untested animals, the presence of vectors, and needle sharing by owners. The data allowed defining Tapira as an area of expansion of T. vivax epizootic infections in the state of Minas Gerais.

O Trypanosoma vivax é considerado o mais importante trypanosoma patogênico para bovinos e causa grandes prejuízos na pecuária de corte e leite. Este estudo teve como objetivo avaliar a prevalência anticorpos de anti-Trypanosoma vivax em bovinos leiteiros do município de Tapira, localizado na região do Alto Paranaíba, Minas Gerais, Brasil. As 74 amostras de soro sanguíneo de bovinos leiteiros foram analisadas por meio de reação de imunofluorescência indireta. A soroprevalência foi de 82,4% (61/74), que pode estar relacionada ao trânsito de animais não testados, presença de vetores e compartilhamento de agulhas pelos proprietários. Os dados permitiram definir Tapira como uma área de expansão das infecções epizoóticas por Trypanosoma vivax no estado de Minas Gerais.

Evaluation of techniques for diagnosis of Trypanosoma vivax infections in naturally infected cattle in the Zona da Mata Mineira / Avaliação de técnicas para diagnóstico de infecções por Trypanosoma vivax em bovinos naturalmente infectados na Zona da Mata Mineira

Abstract This study aimed to evaluate diagnostic techniques for trypanosomiasis, caused by Trypanosoma vivax, in naturally infected cattle in Minas Gerais, Zona da Mata. The deaths of six lactating cows with similar clinical conditions—characterized by hyporexia, hypogalactia, and recumbency—had been reported from one property. Initially, two animals were examined and diagnosed with trypanosomiasis through identification of the protozoan in a blood smear. After the initial diagnosis, all lactating cows (n=37) on the property were examined, and blood samples were collected for tests including whole blood smear, buffy coat smear, Woo's technique, enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction (PCR). Woo's test, buffy coat smears, and whole blood smears indicated that 4/37 (10.81%) animals were positive for trypanosomiasis, whereas ELISA and PCR indicated that 33/37 (89.19%) and 27/37 (72.97%) animals, respectively, were positive. The agreement obtained between parasitological techniques was classified as high, while between ELISA and PCR, no agreement. In conclusion, parasitological techniques have a low capacity to identify infected animals in the chronic stage of T. vivax infection. Therefore, techniques such as PCR and/or ELISA should be used to minimize the occurrence of false negatives.

Resumo Este estudo objetiva avaliar as técnicas de diagnóstico da tripanossomíase, causada pelo Trypanosoma vivax, em bovinos naturalmente infectados, em Minas Gerais, Zona da Mata. A morte de seis vacas em lactação com condições clínicas semelhantes - caracterizadas por hiporexia, hipogalaxia e decúbito - foi relatada em uma propriedade. Inicialmente, dois animais foram examinados e diagnosticados com tripanossomíase através da identificação do protozoário em esfregaço sanguíneo. Após o diagnóstico inicial, todas as vacas em lactação (n = 37) na propriedade foram examinadas, e amostras de sangue foram coletadas para testes, incluindo esfregaço de sangue total, esfregaço de capa leucocitária, técnica de Woo, ensaio imunoenzimático (ELISA) e reação em cadeia da polimerase (PCR). O teste de Woo, os esfregaços de capa leucocitária e de sangue total indicaram que 4/37 (10,81%) animais foram positivos para tripanossomíase, enquanto ELISA e PCR indicaram que 33/37 (89,19%) e 27/37 (72,97%) animais, respectivamente, foram positivos. A concordância entre técnicas parasitológicas foi classificada como alta, enquanto entre ELISA e PCR, sem concordância. As técnicas parasitológicas apresentam baixa capacidade para identificar animais infectados na fase crônica da infecção por T. vivax. Dessa forma, técnicas como PCR e/ou ELISA devem ser utilizadas para minimizar a ocorrência de falsos negativos.

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

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

Prevalence and risk factors for trypanosome infection in cattle from communities surrounding the Murchison Falls National Park, Uganda.

BACKGROUND: Bovine trypanosomosis transmitted by tsetse flies is a major constraint to cattle health and productivity in all sub-Saharan countries, including Uganda. The objectives of this study were to determine the prevalence of bovine trypanosomosis and identify its associated risk factors and the species of trypanosomes associated with the disease. METHODOLOGY: A cross-sectional study was conducted around Murchison Falls National Park, Uganda from January 2020 to April 2020. Trypanosomes were detected in blood samples by PCR analysis targeting the internal transcribed spacer 1 (ITS-PCR assays), and trypanosomes in positive blood samples were sequenced. RESULTS: Of 460 blood samples collected and tested, 136 (29.6%) were positive for trypanosome infections and 324 (70.4%) were negative. The overall trypanosome prevalence was 29.6% (95% confidence interval 25.4-33.8%), attributed to three trypanosome species. Of these three species, Trypanosoma vivax was the most prevalent (n = 130, 28.3%) while the others were detected as mixed infections: T. vivax + Trypanosoma congolense (n = 2, 0.4%) and T. vivax + Trypanosoma evansi (n = 1, 0.2%). There were significant differences in trypanosome prevalence according to sex (χ2 = 62, df = 1, P < 0.05), age (χ2 = 6.28, df = 2, P = 0.0043) and cattle breed (χ2 = 10.61, df = 1, P = 0.001). CONCLUSIONS: Trypanosomosis remains a major limitation to cattle production around Murchison Falls National Park and interventions are urgently needed. In our study, the prevalence of trypanosome infections was high, with T. vivax identified as the most prevalent species. Age, sex and breed of cattle were risk factors for trypanosome infection.

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

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

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

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

New detection of the occurrence of trypanosomiasis in a bovine herd in a rural property in the state of Bahia/BA ­ case report / Nova detecção da ocorrência de tripanossomíase em rebanho bovino em propriedade rural no estado da Bahia/BA ­ relato de caso

This report aims to discuss the occurrence of protozoan infestation in a Girolanda cattle herd in the state of Bahia / BA. Bovine trypanosomiasis is a severe disease that is harmful to cattle, inducing large productive and economic losses. The animals are affected by protozoa of the genus Trypanosoma spp., giving rise to infestations in the vast majority of them in a silent and lasting way. In the Bahian rural property described, 10% of adult cows died and the rest of the animals showed clinical signs of anemia, drop in milk production, low body score, apathy, among others. Among the information obtained through the farm manager, who helped in the diagnosis and in ordering tests for the respective disease were: purchase of cattle from the region of the state of Minas Gerais and dubious management of oxytocin for improvements in milk production (suspected shared needles). Faced with the suspicion and in conjunction with the information obtained, laboratory tests and molecular tests were carried out to confirm the suspicion. After the examinations, it was found that the animals were affected by protozoa of the genus Trypanossoma vivax. The animals were treated with volume replenishers and drugs such as isometamidium chloride in a single dose calculated according to the body weight.

O presente relato tem por objetivo discorrer sobre a ocorrência da infestação protozoária em um rebanho bovino da raça Girolanda no estado da Bahia/BA. A tripanossomíase bovina é uma doença severa e prejudicial aos bovinos, induzindo-os a grandes perdas produtivas e econômicas. Os animais são afetados por protozoários do gênero Trypanosoma spp., dandoorigem a infestações em sua grande maioria de forma silenciosa e duradoura. Na propriedade rural baiana descrita, 10% das vacas adultas morreram e o restante dos animais apresentava sinais clínicos de anemia, queda da produção leiteira, baixo escore corpóreo, apatia, prostração, entre outros. Dentre as informações obtidas através do gerente da fazenda, que auxiliaram no diagnóstico e na requisição de exames para a respectiva doença estavam: compra de bovinos da região do estado de Minas Gerais e manejo duvidoso de ocitocina para melhorias na produção leiteira (suspeita de agulhas compartilhadas). Diante da suspeita e em conjunto com informações obtidas, foram realizados exames laboratoriais e testes moleculares para confirmar a suspeita. Após a realização dos exames, constatou-se que os animais estavam acometidos por protozoários do gênero Trypanossoma vivax. Os animais foram tratados com repositores de volume e fármacos como o cloreto de isometamidium em uma única dose calculada de acordo com o peso corpóreo.

An update on the distribution of Glossina (tsetse flies) at the wildlife-human-livestock interface of Akagera National Park, Rwanda.

BACKGROUND: Glossina (tsetse flies) biologically transmit trypanosomes that infect both humans and animals. Knowledge of their distribution patterns is a key element to better understand the transmission dynamics of trypanosomosis. Tsetse distribution in Rwanda has not been well enough documented, and little is known on their current distribution. This study determined the current spatial distribution, abundance, diversity, and seasonal variations of tsetse flies in and around the Akagera National Park. METHODS: A longitudinal stratified sampling following the seasons was used. Biconical traps were deployed in 55 sites for 6 consecutive days of each study month from May 2018 to June 2019 and emptied every 48 h. Flies were identified using FAO keys, and the number of flies per trap day (FTD) was used to determine the apparent density. Pearson chi-square (χ2) and parametrical tests (t-test and ANOVA) were used to determine the variations between the variables. The significance (p < 0.05) at 95% confidence interval was considered. Logistic regression was used to determine the association between tsetse occurrence and the associated predictors. RESULTS: A total of 39,516 tsetse flies were collected, of which 73.4 and 26.6% were from inside Akagera NP and the interface area, respectively. Female flies accounted for 61.3 while 38.7% were males. Two species were identified, i.e. G. pallidipes [n = 29,121, 7.4 flies/trap/day (FTD)] and G. morsitans centralis (n = 10,395; 2.6 FTD). The statistical difference in numbers was significant between the two species (p = 0.000). The flies were more abundant during the wet season (15.8 FTD) than the dry season (4.2 FTD). Large numbers of flies were trapped around the swamp areas (69.1 FTD) inside the park and in Nyagatare District (11.2 FTD) at the interface. Glossina morsitans was 0.218 times less likely to occur outside the park. The chance of co-existing between the two species reduced outside the protected area (0.021 times). CONCLUSIONS: The occurrence of Glossina seems to be limited to the protected Akagera NP and a narrow band of its surrounding areas. This finding will be crucial to design appropriate control strategies. Glossina pallidipes was found in higher numbers and therefore is conceivably the most important vector of trypanosomosis. Regional coordinated control and regular monitoring of Glossina distribution are recommended.

A recombinant protein (MyxoTLm) for the serological diagnosis of acute and chronic Trypanosoma vivax infection in cattle.

Human trypanosomiases and animal trypanosomoses are caused by distinct protozoan parasites of the genus Trypanosoma. The etiological agents of bovine trypanosomosis (BT) are T. vivax, T. congolense, or T. brucei, whose acute infections are initially characterized by hyperthermia, following moderate to severe anemia, subcutaneous edema, lethargy, reduced milk production, progressive weight loss, enlarged lymph nodes, reproductive disorders and death. Animals that survive the acute phase might recover and progress to the chronic, often asymptomatic, phase of infection. Despite their low sensitivity due to the characteristic low parasitemia, simple and costless direct parasitological examinations are the preferred diagnostic methods for animals. Thus, most of the epidemiological studies of BT are based on serological techniques using crude antigen. In this study, we describe the use of the MyxoTLm recombinant protein as an antigen on serological assays. Anti-T. vivax IgM and anti-T. vivax IgG ELISA assays using purified MyxoTLm revealed specificity rates of 91.30 % and 95.65 % and sensitivity rates of 82.35 % and 88.23 %, respectively, being higher than reported for crude antigens. Also, MyxoTLm demonstrated a good performance to detect IgM (ROC curve area = 0.8568) and excellent performance to detect IgG (ROC curve area = 0.9565) when compared to a crude antigen. T. evansi crude antigen used in the indirect anti-T. vivax IgM ELISA reached 70.58 % sensitivity and 78.26 % specificity, and had a lower test performance (ROC curve area = 0.7363). When applied to the anti-T. vivax IgG ELISA, the crude antigen reached 82.35 % sensitivity and 69.56 % specificity, also presenting a low performance with area under the ROC curve of 0.7570. Therefore, the use of MyxoTLm as an antigen on serological diagnosis of BT revealed to increase the sensitivity and the specificity if compared to crude antigens.