African Animal Trypanosomosis Research Articles

Immune Response in Cattle Trypanosomosis and Trypanotolerance: Main Findings and Gaps.

Trypanosome parasites of the genus Trypanosoma cause African animal trypanosomosis, a devastating livestock disease plaguing sub-Saharan Africa. Unlike many protozoan parasites, these extracellular blood-borne pathogens directly engage the host's immune system. While the mouse model has provided valuable insights, a comprehensive understanding of the bovine immune response to trypanosomes remains elusive. Addressing the immune response in cattle, the most relevant host species, and how it takes part in mitigating the negative impact of the disease could contribute to setting up sustainable control strategies. This review summarises the current knowledge of the immune response in cattle during trypanosomosis. Following a brief overview of infection processes and bovine trypanotolerance, we present advances in the regulation of host innate, inflammatory and adaptive responses and delve into the key immunological players involved in immunoactivities and immunosuppression. We discuss how these mechanisms contribute to tolerance or susceptibility to infection, highlighting critical gaps in knowledge that require further investigation.

Bovine trypanosomosis, vector distribution and infection rate in three districts of Gamo Zone, southwestern Ethiopia

African animal trypanosomosis is one of the main obstacles to the development of livestock and agricultural output in Ethiopia. It usually results in a severe, frequently fatal sickness, and the infected animals were more weakened as the disease progress and become unfit for work. A cross sectional study design was conducted from December 2021 to April 2022 with the aim of estimating the prevalence of trypanosome infection both in Glossina spp. and cattle, and to assess apparent density of Glossina spp. A total of 298 cattle were selected and examined for trypanosome by using buffy coat technique. The overall prevalence of bovine trypanosomosis was 19.1%; and two species of trypanosomes, T. congolense and T. vivax, were identified in the study area. The prevalence of T. congolense and T. vivax were 15.8% and 2.3%, respectively. The prevalence of trypanosomosis was significantly higher in adult animals (OR = 2.7; p < 0.05) than in younger cattle and poor body condition (OR = 3.18; p < 0.05) than medium body condition animals. The mean PCV value of infected animals was 14.3% (13.3–15.4) significantly lower than the non-infected animals 18.5% (17.8–19.2). Glossina pallidipes is the only tsetse species encountered in all the study areas. In total, 2992 flies were caught of which 90.8% belong to G. pallidipes and 9.2% were other biting flies. The overall apparent density of G. pallidipes was 20.1 F/T/D and other biting flies were 2.0 F/T/D. A total of 307 live Glossina pallidipes were dissected. The overall prevalence of Glossina pallidipes infection rate was 9.1% (95% CI = 5.9–12.4). The prevalence of G. pallidipes infection was significantly higher in Kucha district (OR = 3.2, χ2 = 2.6, p < 0.05) than the other two districts, Daramalo and Arba Minch Zuria. Also it was significantly higher in flies trapped from riverine forest areas (OR = 5.5, χ2 = 2.86, p < 0.05). Therefore, to reduce the impact of trypanosomosis and Glossina, vector control and treating infected cattle with prophylactic or chemotherapeutic drugs and active community participation can play a key role.

An Overview of Tsetse Fly Repellents: Identification and Applications.

Tsetse flies are vectors of the parasite trypanosoma that cause the neglected tropical diseases human and animal African trypanosomosis. Semiochemicals play important roles in the biology and ecology of tsetse flies. Previous reviews have focused on olfactory-based attractants of tsetse flies. Here, we present an overview of the identification of repellents and their development into control tools for tsetse flies. Both natural and synthetic repellents have been successfully tested in laboratory and field assays against specific tsetse fly species. Thus, these repellents presented as innovative mobile tools offer opportunities for their use in integrated disease management strategies.

Assessment of optimal blood exposure time at 37 °C during in vitro tsetse flies feeding for quality production in mass-rearing colonies.

Control of African animal trypanosomosis is implemented through an integrated control strategy, with the sterile insect technique (SIT) as one of its components. The SIT requires mass rearing of tsetse fly colonies using an in vitro feeding system. The exposure of blood at 37 °C on heating plates over time can have an impact on the quality of fly productivity. In this study, we investigated the survival and fecundity of adult tsetse flies fed at 37 °C on 8 blood exposure times ranging from 30 min to 4 h with increments of 30 min (treatment 1, flies were fed 30 min after exposure to blood at 37 °C; treatment 2, 1 h and so on until treatment 8 [4 h after]) in order to determine the optimal exposure time. In addition, bacterial growth in blood from each treatment was assessed by agar culture at 37 °C for 72 h. The results showed that the adult female survival rates were similar regardless of the treatment. For males, only those of treatment 1 (30 min) showed a marginal lower survival than those of treatments 7 and 8 fed after 3 h 30 min and 4 h of blood exposure, respectively. Over the 4-h interval of blood exposure at 37 °C, the results showed that the number of pupae produced per initial female and pupal weight tended to increase with exposure time, but the differences were not significant. We discuss the implications of these results on tsetse mass rearing for the SIT program.

Prophylactic activity of orally administered dry-heat-sterilized Acremonium egyptiacum against Trypanosoma congolense-induced animal African trypanosomosis

Animal African trypanosomosis (AAT) is an important global disease of livestock that causes economic losses of up to 4.5 billion US dollars per year. Thus, eliminating AAT in endemic countries will improve agricultural productivity and economic growth. To prevent AAT, vector control and the development of prophylactic drugs are crucial. Ascofuranone (AF) is a bioactive fungal compound with proven in vitro trypanocidal potency and in vivo treatment efficacy. However, the complex stereoselective synthesis of AF has prevented its cost-effective industrial production. Recently, a genetically modified strain of Acremonium egyptiacum fungus that produces a high yield of AF was developed. Therefore, we hypothesized that the oral administration of the AF-producing fungus itself may be effective against AAT. Hence, this study aimed to evaluate the prophylactic activity of orally administered dry-heat-sterilized A. egyptiacum against Trypanosoma congolense IL3000 infection using a mouse model. The survival rate was significantly prolonged (p = 0.009), and parasitemia was suppressed in all AF-fungus-treated groups (Group 1–9) compared with that in the untreated control group (Group 10). Hence, the trypanocidal activity of AF was retained after dry-heat-sterilization of the AF-producing fungus and that its oral administration effectively prevented AAT. Since AAT is endemic to rural areas with underdeveloped veterinary infrastructure, dry-heat-sterilized A. egyptiacum would be the most cost-effective potential treatment for AAT.

In vitro antitrypanosomal activity of synthesized nitrofurantoin-triazole hybrids against Trypanosoma species causing animal African trypanosomosis

Animal African trypanosomosis (AAT) is a disease caused by Trypanosoma brucei brucei, T. vivax, T. evansi and T. congolense which are mainly transmitted by tsetse flies (maybe the family/genus scientific name for the tsetse flies here?). Synthetic trypanocidal drugs are used to control AAT but have reduced efficacy due to emergence of drug resistant trypanosomes. Therefore, there is a need for the continued development of new safe and effective drugs. The aim of this study was to evaluate the in vitro anti-trypanosomal activity of novel nitrofurantoin compounds against trypanosomes (Trypanosoma brucei brucei, T. evansi and T. congolense) causing AAT. This study assessed previously synthesized nineteen nitrofurantoin-triazole (NFT-TZ) hybrids against animal trypanosomes and evaluated their cytotoxicity using Madin–Darby bovine kidney cells. The n-alkyl sub-series hybrids, 8 (IC50 0.09 ± 0.02 μM; SI 686.45) and 9 (IC50 0.07 ± 0.04 μM; SI 849.31) had the highest anti-trypanosomal activity against T. b. brucei. On the contrary, the nonyl 6 (IC50 0.12 ± 0.06 μM; SI 504.57) and nitrobenzyl 18 (IC50 0.11 ± 0.03 μM; SI 211.07) displayed the highest trypanocidal activity against T. evansi. The nonyl hybrid 6 (IC50 0.02 ± 0.01 μM; SI 6328.76) was also detected alongside the undecyl 8 (IC50 0.02 ± 0.01 μM; SI 3454.36) and 3-bromobenzyl 19 (IC50 0.02 ± 0.01 μM; SI 2360.41) as the most potent hybrids against T. congolense. These hybrids had weak toxicity effects on the mammalian cells and highly selective submicromolar antiparasitic action efficacy directed towards the trypanosomes, hence they can be regarded as potential trypanocidal leads for further in vivo investigation.

Animal trypanosomosis eliminated in a major livestock production region in Senegal following the eradication of a tsetse population.

African animal trypanosomosis (AAT) was one of the main disease-related constraints to the development of intensive livestock production systems in the Niayes region of Senegal, a 30km wide strip of land along the coast between Dakar and Saint-Louis. To overcome this constraint, the Government of Senegal initiated an area-wide integrated pest management programme combining chemical control tactics with the sterile insect technique to eradicate a population of the tsetse fly Glossina palpalis gambiensis Vanderplank, 1949 (Diptera, Glossinidae) in this area. The project was implemented following a phased conditional approach, and the target area was divided into three blocks treated sequentially. This study aims to assess the temporal dynamics of the prevalence of Trypanosoma spp. during the implementation of this programme. Between 2009 and 2022, 4,359 blood samples were collected from cattle and screened for trypanosomes using both the buffy coat and ELISA techniques, and PCR tests since 2020. The seroprevalence decreased from 18.9% (95%CI: 11.2-26.5) in 2009 to 0% in 2017-2022 in block 1, and from 92.9% (95%CI: 88.2-97) in 2010 to 0% in 2021 in block 2. The parasitological and serological data confirm the entomological monitoring results, i.e., that there is a high probability that the population of G. p. gambiensis has been eradicated from the Niayes and that the transmission of AAT has been interrupted in the treated area. These results indicate the effectiveness of the adopted approach and show that AAT can be sustainably removed through the creation of a zone free of G. p. gambiensis.

Ethno-Botanical Survey: Exploring Indigenous Remedies for African Animal Trypanosomosis in Nigeria

The Fulani pastoralist of Nigeria and other indigenous livestock farmers have used medicinal plants to treat various animal diseases for centuries, amongst which include the African animal trypanosomosis (AAT). It is a severe debilitating parasitic disease of livestock and some wild animal species. It is caused by pathogenic Trypanosomes and transmitted by Tsetse flies. It causes severe economic losses to livestock farmers in Africa. In this study, an ethno-botanical survey was conducted in the Tsetse-endemic areas of Bauchi, Plateau and Kaduna States of Nigeria to identify medicinal plants used traditionally to treat AAT. During the study, eighty questionnaires were administered to the respondents who were predominantly Fulani pastoralists, and other indigenous livestock farmers. seventy-eight questioners were fully completed and retrieved. The study revealed that that most of the respondents have retained the tradition knowledge system of medicinal plants passed down to them for generations. The study revealed various herbal preparation methods employed by indigenous Nigerian livestock farmers, including decoctions, infusions, and incorporating plant material into feed. Fifteen medicinal plants species belonging to 13 families were identified to be used by Nigerian pastoralist for the treatment AAT. These included Khaya senegalensis (28.2%), Azadirachta indica (25.6%), Boswellia dalziellii (23%), Prosopis africana (23%), Anogeisus leocarpus and Terminalia avicennioides (21.8%), Adansonia digitata and Annona senegalensis (19.2%), Pilostigma thoningii (16.7%), Nauclea latifolia (15.0%). Balanites aegyptiaca (12.5%). Eucalyptus torelliana (11.5%), Tamarindus indica (10.3%), Ximenia americana (9%) and Guiera senegalensis 7.7%).&#x0D; Conclusions: The information obtained from this study and the medicinal plants identified can be harnessed for the development of plant-based medicaments for the treatment of AAT.

In Vitro Trypanocidal Effect of the Extract and Isolated Compound of Corymbia torelliana Stem Bark against Trypanosoma brucei brucei (Federe Strain)

African animal trypanosomosis (AAT) is a severe debilitating protozoan disease of livestock and some wild animal species. It is caused by the pathogenic trypanosome species, and transmitted by Tsetse flies. It affects about 60 million cattle in Africa, with about US$1.5 billion in annual losses. Eradication of the disease through chemotherapy and vaccine development has been hindered by parasite antigenic variation and its resistance to drugs. In this study, the crude ethanol stem bark extract of Corymbia torelliana was prepared by cold maceration in 70% Ethanol. It was further purified using the modified Kupchan partition method into n-hexane, Ethyl acetate, N-butanol and Aqueous pure fractions. A pure compound was isolated from the ethyl acetate purified fraction using the column and thin layer chromatographic techniques. The isolated compound was named Makoshi_Mak1. The in vitro antitrypanosomal effects of the crude extract, the purified fractions and the isolated compound were then tested against the blood stream forms of Trypanosoma brucei brucei (Federe strain). 0.2 ml of prepared trypanosome suspension containing about 2x104 parasites was dispensed into each of seven clean test tubes using micropipettes. 0.2ml of prepared Ringer’s solution was also added to stabilize the parasites. Five varying concentrations of the crude extract, the extract fractions and the isolated compound (Makoshi_Mak1) were added to test tubes 1-5. Test tube 6 and 7 served as the negative and positive controls respectively. Aliquots of the contents of each test tube were examined under the microscope at x400 magnification for parasites motility at different time interval for 2 hours. The crude extract was effective at the concentrations of 12 mg/ml, 1.2 mg/ml,0.12 mg/ml and 0.012 mg/ml. the n-hexane fraction had efficacy at 0.12mg/ml and 0.096mg/ml. The Eethyl acetate fractions were effective at 0.12mg/ml, 0.096 mg/ml, 0.048mg/ml and 0.024mg/ml, the N-butanol fraction had efficacy at 0.12mg/ml and 0.096mg/ml, while the Aqueous fraction was effective at 0.2mg/ml, 0.16mg/ml and 0.08mg/ml. Makoshi_Mak1 was effective in vitro against T. brucei beucei at all the tested concentrations of 0.219mg/ml, 0.109mg/ml, 0.055mg/ml and 0.027mg/ml. Efficacy of the extracts and isolated compound was determined by parasite mortality, and was seen to be concentration dependent. Dead parasites appeared to float freely on the fluid environment, while the lysed parasites were seen as floating debris. The results of the study showed that both the crude extract, the purified extract and the pure isolated compound of Corymbia torelliana had in vitro activity against Trypanosoma brucei brucei in a concentration dependent manner. Corymbia torelliana can then be considered as a potential herbal antitrypanosomal drug candidate.

Performance of diagnostic tests for Trypanosoma brucei brucei in experimentally infected pigs.

Animal African trypanosomosis is an important vector-borne disease of livestock in sub-Saharan Africa. Pigs seem relatively tolerant to trypanosome infection and could act as a reservoir of trypanosomes affecting animals and humans. Our ability to reliably detect trypanosome infection in pigs depends on the performance of diagnostic tools, which is not well known. In pigs experimentally infected with Trypanosoma brucei brucei, we evaluated the performance of parasitological Buffy Coat Technique (BCT), two molecular (TBR and 5.8S PCR) and four serological tests (CATT, HAT Sero-K-Set rapid diagnostic test-RDT, indirect ELISA, immune trypanolysis). Most diagnostic tests showed high specificity, estimated at 100% (95% CI = 74-100%) with the exception of CATT and RDT whose specificity varied between 100% (95% CI = 74-100%) to 50% (95% CI = 7-93%) during the experiment. The sensitivity of each test fluctuated over the course of the infection. The percentage of positive BCT over the infection (30%) was lower than of positive PCR (56% and 62%, depending on primers). Among the serological tests, the percentage of positive tests was 97%, 96%, 86% and 84% for RDT, ELISA, immune trypanolysis and CATT, respectively. Fair agreement was observed between both molecular tests (κ = 0.36). Among the serological tests, the agreement between the ELISA and the RDT was substantial (κ = 0.65). Our results on the T.b. brucei infection model suggest that serological techniques are efficient in detecting the chronic phase of infection, PCR is able to detect positive samples several months after parasites inoculation while BCT becomes negative. BCT examination and RDT are useful to get a quick information in the field, and BCT can be used for treatment decision. ELISA appears most suited for epidemiological studies. The selection of diagnostic tests for trypanosomosis in pigs depends on the context, the objectives and the available resources.

Diversity of trypanosome species in small ruminants, dogs and pigs from three sleeping sickness foci of the south of Chad.

Despite considerable data generated on livestock trypanosomoses in tsetse-infested areas, little attention was paid for animal African trypanosomosis (AAT) in sleeping sickness foci. This study aimed to fill this gap by determining the diversity and prevalence of trypanosome species in animals from three Chadian human African trypanosomosis (HAT) foci. Blood samples were collected from 443 goats, 339 sheep, 228 dogs and 98 pigs of the Mandoul, Maro and Moissala HAT foci in the south of Chad. Capillary tube centrifugation (CTC) and specific primers were used to search trypanosomes. The prevalence of trypanosome infections was 6.3% for CTC and 22.7% for PCR. Trypanosomes of the sub-genus Trypanozoon had the highest prevalence (16.6%) while T. congolense savannah (1.9%) was least prevalent. Significant differences were recorded between the prevalence of trypanosome species (χ2 = 8.34; p = 0.04) and HAT foci (χ2 = 24.86; p ≤0.0001). Maro had the highest prevalence (32.7%) and Mandoul the lowest (17.4%). Significant differences were also recorded for T. congolense forest (χ2 = 45.106; p < 0.0001) and all T. congolense (χ2 = 34.992; p < 0.0001). Goats had the highest prevalence (26.9%) and sheep the lowest one (18.6%). Between animals, significant differences were recorded for trypanosome of the sub-genus Trypanozoon (χ2 = 9.443; p = 0.024), T. congolense forest (χ2 = 10.476; p = 0.015) and all T. congolense (χ2 = 12.152; p = 0.007). Of the 251 animals carrying trypanosome infections, 88.8% had single infections while 11.2% had more than one trypanosome species. The overall prevalence of single and mixed trypanosome infections were respectively 20.1% and 2.6% in animal taxa from of all foci. This study highlighted a diversity of trypanosomes in animal taxa of all HAT foci. It showed that AAT constitutes a threat for animal health and animal breeding in Chadian HAT foci. In these tsetse infested areas, reaching the elimination of AAT requires the designing and the implementation of control measures against trypanosome infections.

Recent advances in genome editing of bloodstream forms of Trypanosoma congolense using CRISPR-Cas9 ribonucleoproteins: Proof of concept

African Animal Trypanosomosis (AAT or Nagana) is a vector-borne disease caused by Trypanosomatidae, genus Trypanosoma. The disease is transmitted by the bite of infected hematophagous insects, mainly tsetse flies but also other blood-sucking insects including stomoxes and tabanids. Although many trypanosome species infect animals, the main agents responsible for this disease with a strong socio-economic and veterinary health impact are Trypanosoma congolense (T. congolense or Tc), Trypanosoma vivax (T.vivax), and to a lesser extent, Trypanosoma brucei brucei (T.brucei brucei or Tbb). These parasites mainly infect livestock, including cattle, in sub-Saharan Africa, with major repercussions in terms of animal productivity and poverty for populations which are often already very poor. As there is currently no vaccine, the fight against the disease is primarily based on diagnosis, treatment and vector control. To develop new tools (particularly therapeutic tools) to fight against the disease, we need to know both the biology and the genes involved in the pathogenicity and virulence of the parasites. To date, unlike for Trypanosoma brucei (T.brucei) or Trypanosoma cruzi (T.cruzi), genome editing tools has been relatively little used to study T. congolense. We present an efficient, reproducible and stable CRISPR-Cas9 genome editing system for use in Tc bloodstream forms (Tc-BSF). This plasmid-free system is based on transient expression of Cas9 protein and the use of a ribonucleoprotein formed by the Cas9 and sgRNA complex. This is the first proof of concept of genome editing using CRISPR-Cas9 ribonucleoproteins on Tc-BSF. This adapted protocol enriches the “toolbox” for the functional study of genes of interest in blood forms of the Trypanosoma congolense. This proof of concept is an important step for the scientific community working on the study of trypanosomes and opens up new perspectives for the control of and fight against animal trypanosomosis.

Efficacy of oral administration of ascofuranone with and without glycerol against Trypanosoma congolense

In many developing countries, trypanosomosis in animals results in the reduction of livestock productivity. Since trypanosomosis is endemic to rural areas where medical and veterinary infrastructure is underdeveloped, development of affordable and easy-to-maintain drugs for treatment and prophylaxis against trypanosomosis is necessary. To this end, in this study, we evaluated the efficacy of oral administration of ascofuranone (AF), with and without glycerol (GOL), against trypanosomosis, using a mouse model. We used T. congolense IL3000, the most virulent animal-infecting trypanosome, and BALB/c mice in this study. Eight mice were assigned to either of Groups 1–7: non-infected, untreated, AF 10, 20, 30, 50, and 100 mg/kg with or without GOL, respectively. In the experiment with AF administered with GOL, survival rates were 0% in Group 2 (untreated) and Group 3 (AF 10 mg/kg), 37.5% in Group 4 (AF 20 mg/kg) and Group 5 (AF 30 mg/kg), 50% in Group 6 (AF 50 mg/kg), and 100% in Group 7 (AF 100 mg/kg). In groups in which AF was administered without GOL, survival rates were 0% in Group 2 (untreated), Group 3 (AF 10 mg/kg), Group 4 (AF 20 mg/kg), Group 5 (AF 30 mg/kg), and Group 6 (AF 50 mg/kg), and 12.5% in Group 7 (AF 100 mg/kg), with one mouse surviving till the end of the observation period. The results of the analysis showed that survival rates were significantly higher in all groups (Groups 3–7) than in the untreated group (Group 2) (p < 0.05). Furthermore, a comparison of groups with or without GOL at the same AF concentration revealed that the survival rate was significantly higher in the group treated with GOL. These results suggest that the treatment efficacy of AF against animal trypanosomosis caused by T. congolense is greater when co-administered with GOL, and that oral administration of AF could be a new therapeutic strategy for animal African trypanosomosis.

Surveillance and control of Trypanosoma evansi in the canary Islands: A descriptive analysis

This study examines the occurrence of Surra, a disease caused by Trypanosoma evansi, in camels in the Canary Islands. The 1997 detection of T. evansi in camels in the Canary Islands led to the implementation of an initial control program, resulting in a decrease in prevalence. Following an outbreak in 2014, and due to the impossibility of eradicating it using the conventional measures, a lazaret was set up to separate positive and suspicious animals, in addition to the control measures previously implemented. Stomoxys calcitrans was the only vector captured, and no other animals tested were found to be positive for T. evansi. In November 2019, the last camels that tested serologically positive were detected; however, since February 2018, no camels that tested positive for PCR have been found in the farms were the outbreak was detected, suggesting that the sanitary measures implemented are adequate. The duration of the outbreak control and potential eradication for the disease has yet to be established. This study provides evidence to facilitate the control of African Animal Trypanosomosis in endemic areas of the world, which may contribute to revise the World Organization for Animal Health (WOAH) protocol to implement recommendations of surveillance and control strategies for animal Trypanosomosis in camels.

Population genetics of Glossina fuscipes fuscipes from southern Chad

In Subsaharan Africa, tsetse flies (genus Glossina) are vectors of trypanosomes causing Human African Trypanosomiasis (HAT) and Animal African Trypanosomosis (AAT). Some foci of HAT persist in Southern Chad, where a program of tsetse control was started against the local vector Glossina fuscipes fuscipes in the Mandoul focus in 2014, and in Maro in 2018. Flies were also sampled in 2018 in Timbéri and Dokoutou. We analyzed the population genetics of G. fuscipes fuscipes from the four tsetse-infested zones. The trapping samples were characterized by a strong female biased sex-ratio, except in Timbéri and Dokoutou that had high tsetse densities. Apparent density and effective population density appeared smaller in the main foci of Mandoul and Maro and the average dispersal distance (within the spatial scale of each zone) was as large as or larger than the total length of each respective zone. The genetic signature of a population bottleneck was found in the Mandoul and Timbéri area, suggesting a large ancient interconnected metapopulation that underwent genetic subdivision into small, isolated pockets due to adverse environmental conditions. The long-range dispersal and the existence of genetic outliers suggest a possibility of migration from remote sites such as the Central African Republic in the south (although the fly situation remains unknown there) and/or a genetic signature of recent exchanges. Due to likely isolation, an eradication strategy may be considered for sustainable HAT control in Mandoul focus. Another strategy will probably be required in Maro focus, which probably experiences much more exchanges with its neighbors.

Feeding frequency and its associated effects on the production and survival rate of Glossina fuscipes fuscipes

Tsetse flies are large blood-sucking flies of the genus Glossina, which are exclusively found in Africa. They are important vectors of Trypanosomes, which cause sleeping sickness in humans and African Animal Trypanosomosis (AAT) in livestock. Due to the lack of effective vaccines to control this disease, vector control has been an ideal strategy. There are many vector control methods developed so far. Sterile Insect Technique (SIT) is one of such method which depends on the mass production of male flies in the laboratory, sterilization, and release of these flies in the target area. The study was conducted to evaluate the effect of different feeding frequencies on the production and survival rate of Glossina fuscipes fuscipes to identify the best feeding frequency that could optimize the production and minimize the cost of production in a massrearing insectary facility at Kaliti Tsetse Fly Mass Rearing and Irradiation Centre, Addis Ababa, Ethiopia. Three experimental groups were established based on feeding frequency regimes. The first group was fed three times per week, the second four times per week, and the third group five times per week. Parameters that are routinely used for assessing colony performance such as; survival rate, fecundity, pupae per initial females (PPIF), and pupal quality were evaluated for each treatment group and statistical comparisons were made between the groups. The results revealed that feeding frequency had a crucial effect on fly production and survival. The lowest results for all parameters were obtained in flies fed three times per week and relatively the best results were obtained in the four-times-per-week feeding regime. Therefore, we recommend a further study on aspects that are not addressed in this study, such as emergency rate and progeny quality, to apply the four times per week feeding regime as it will have a positive economic impact that could enable more resources to be re-injected into the insectary compared to five or more feeding regimes which are currently in use in the study site.

Vector competence of sterile male Glossina fuscipes fuscipes for Trypanosoma brucei brucei: implications for the implementation of the sterile insect technique in a sleeping sickness focus in Chad

BackgroundHuman African trypanosomiasis (HAT) is a neglected tropical disease caused by Trypanosoma brucei gambiense transmitted by tsetse flies in sub-Saharan West Africa. In southern Chad the most active and persistent focus is the Mandoul focus, with 98% of the reported human cases, and where African animal trypanosomosis (AAT) is also present. Recently, a control project to eliminate tsetse flies (Glossina fuscipes fuscipes) in this focus using the sterile insect technique (SIT) was initiated. However, the release of large numbers of sterile males of G. f. fuscipes might result in a potential temporary increase in transmission of trypanosomes since male tsetse flies are also able to transmit the parasite. The objective of this work was therefore to experimentally assess the vector competence of sterile males treated with isometamidium for Trypanosoma brucei brucei.MethodsAn experimental infection was set up in the laboratory, mimicking field conditions: the same tsetse species that is present in Mandoul was used. A T. b. brucei strain close to T. b. gambiense was used, and the ability of the sterile male tsetse flies fed on blood with and without a trypanocide to acquire and transmit trypanosomes was measured.ResultsOnly 2% of the experimentally infected flies developed an immature infection (midgut) while none of the flies developed a metacyclic infection of T. b. brucei in the salivary glands. We did not observe any effect of the trypanocide used (isometamidium chloride at 100 mg/l) on the development of infection in the flies.ConclusionsOur results indicate that sterile males of the tested strain of G. f. fuscipes were unable to cyclically transmit T. b. brucei and might even be refractory to the infection. The data of the research indicate that the risk of cyclical transmission of T. brucei by sterile male G. f. fuscipes of the strain colonized at IAEA for almost 40 years appears to be small.Graphical

Nanoformulation of a Trypanocidal Drug Isometamidium Chloride Ameliorates the Apurinic-Apyrimidinic DNA Sites/Genotoxic Effects in Horse Blood Cells.

Isometamidium chloride (ISM) is a trypanocide for the prophylactic and therapeutic use against vector-borne animal trypanosomosis (mainly Surra caused by Trypanosoma evansi) and African animal trypanosomosis caused by T. congolense/T. vivax/T. brucei). ISM was found to be an efficient trypanocide for therapeutic/prophylactic use against trypanosomosis; however, it produces some local and systemic detrimental effects in animals. We synthesized isometamidium chloride-loaded alginate gum acacia nanoformulation (ISM SANPS) to lessen the detrimental side effects of isometamidium chloride (ISM) while treating trypanosomal diseases. We intended to determine the cytocompatibility/toxicity, and DNA deterioration/chromosomal structural or number changes (genotoxicity) of ISM SANPs using mammalian cells in a concentration-dependent manner. Apurinic/apyrimidinic (AP) sites are one of the major types of DNA lesions formed during base excision and repair of oxidized, deaminated, or alkylated bases. The intensity of the cellular AP site is an excellent marker of the deterioration of DNA quality. We thought it pertinent to quantify the AP sites in ISM SANPs-treated cells. Our investigations established a dose-dependent cyto-compatibility or toxicity and DNA impairment (genotoxicity) in ISM SANPs-treated horse peripheral blood mononuclear cells. ISM SANPs were biocompatible at various concentrations tested on the mammalian cells.

Whole blood transcriptome profiles of trypanotolerant and trypanosusceptible cattle highlight a differential modulation of metabolism and immune response during infection by Trypanosoma congolense

Animal African trypanosomosis, caused by blood protozoan parasites transmitted mainly by tsetse flies, represents a major constraint for millions of cattle in sub-Saharan Africa. Exposed cattle include trypanosusceptible indicine breeds, severely affected by the disease, and West African taurine breeds called trypanotolerant owing to their ability to control parasite development, survive and grow in enzootic areas. Until now the genetic basis of trypanotolerance remains unclear. Here, to improve knowledge of the biological processes involved in trypanotolerance versus trypanosusceptibility, we identified bovine genes differentially expressed in five West African cattle breeds during an experimental infection by Trypanosoma congolense and their biological functions. To this end, whole blood genome-wide transcriptome of three trypanotolerant taurine breeds (N’Dama, Lagune and Baoulé), one susceptible zebu (Zebu Fulani) and one African taurine x zebu admixed breed (Borgou) were profiled by RNA sequencing at four time points, one before and three during infection. As expected, infection had a major impact on cattle blood transcriptome regardless of the breed. The functional analysis of differentially expressed genes over time in each breed confirmed an early activation of the innate immune response, followed by an activation of the humoral response and an inhibition of T cell functions at the chronic stage of infection. More importantly, we highlighted overlooked features, such as a strong disturbance in host metabolism and cellular energy production that differentiates trypanotolerant and trypanosusceptible breeds. N’Dama breed showed the earliest regulation of immune response, associated with a strong activation of cellular energy production, also observed in Lagune, and to a lesser extent in Baoulé. Susceptible Zebu Fulani breed differed from other breeds by the strongest modification in lipid metabolism regulation. Overall, this study provides a better understanding of the biological mechanisms at work during infection, especially concerning the interplay between immunity and metabolism that seems differentially regulated depending on the cattle breeds.

Prevalence and Association of Trypanosomes and Sodalis glossinidius in Tsetse Flies from the Kafue National Park in Zambia

Tsetse flies are obligate hematophagous vectors of animal and human African trypanosomosis. They cyclically transmit pathogenic Trypanosoma species. The endosymbiont Sodalis glossinidius is suggested to play a role in facilitating the susceptibility of tsetse flies to trypanosome infections. Therefore, this study was aimed at determining the prevalence of S. glossinidius and trypanosomes circulating in tsetse flies and checking whether an association exists between trypanosomes and Sodalis infections in tsetse flies from Kafue National Park in Zambia. A total of 326 tsetse flies were sampled from the Chunga and Ngoma areas of the national park. After DNA extraction was conducted, the presence of S. glossinidius and trypanosome DNA was checked using PCR. The Chi-square test was carried out to determine whether there was an association between the presence of S. glossinidius and trypanosome infections. Out of the total tsetse flies collected, the prevalence of S. glossinidius and trypanosomes was 21.8% and 19.3%, respectively. The prevalence of S. glossinidius was 22.2% in Glossina morsitans and 19.6% in Glossina pallidipes. In relation to sampling sites, the prevalence of S. glossinidius was 26.0% in Chunga and 21.0% in Ngoma. DNA of trypanosomes was detected in 18.9% of G. morsitans and 21.4% of G. pallidipes. The prevalence of trypanosomes was 21.7% and 6.0% for Ngoma and Chunga, respectively. The prevalences of trypanosome species detected in this study were 6.4%, 4.6%, 4.0%, 3.7%, 3.1%, and 2.5% for T. vivax, T. simiae, T. congolense, T. godfreyi, T. simiae Tsavo, and T. b. brucei, respectively. Out of 63 trypanosome infected tsetse flies, 47.6% of the flies also carried S. glossinidius, and the remaining flies were devoid of S. glossinidius. A statistically significant association was found between S. glossinidius and trypanosomes (p < 0.001) infections in tsetse flies. Our findings indicated that presence of S. glossinidius increases the susceptibility of tsetse flies to trypanosome infections and S. glossinidius could be a potential candidate for symbiont-mediated vector control in these tsetse species.