In vitro and in vivo Antitrypanosomal Activities of Annona muricata Leaf Extracts in Trypanosoma brucei brucei Experimentally Infected Albino Rats

Osho Innocent Bamidele, Durojaye Catherine Olusunmibola
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Abstract

Aims: Trypanosomiasis is an important protozoan disease that affects domestic and wild animals as well as man. It is caused by the tsetse fly-transmitted extracellular hemo-flagellates that belong to the genus, Trypanosoma. In East and Southern Africa, Human African Trypanosomiasis (HAT) is caused by Trypanosoma brucei rhodesiense while in West and Central Africa, it is caused by T. b. gambiense. Animal trypanosomiasis on the other hand is caused by T. b. brucei, T. vivax, and T. congolense. In sub-Saharan Africa, about sixty million people are at risk of infection. This current study evaluates the antitrypanosomal efficacy of extracts of Annona muricata leaf in Trypanosoma brucei brucei infected albino rats. Place and Duration of Study: Animal Parasitology and Microbiology Research Unit, Department of Animal Production and Health, Federal University of Technology, Akure, Nigeria, between February and June, 2021. Methodology: In vitro antitrypanosomal analysis was done in varied concentrations of 2.5mg/ml, 5mg/ml and 10mg/ml using various solvent extracts (ethanolic, ethyl acetate, n-hexane, chloroform and aqueous). Diminazine aceturate and normal saline were used as positive and negative controls respectively. The In vivo assay was carried out through intraperitoneal administration of graded doses (200, 400 and 600mg/kg) of ethyl acetate and chloroform extracts of the plant for three consecutive days. Results: The n-hexane, chloroform and ethyl acetate extracts yielded high percentage DPPH free radical scavenging activity of 85.52, 80.00 and 78.49% respectively. Decrease in motility of the parasites at different times were observed in all extracts tested invitro. These responses were positively concentration-dependent. 10mg/ml concentration of chloroform and ethyl acetate showed complete cessation of parasite motility at 35 and 45 minutes respectively. These two extracts (ethyl acetate and chloroform extracts) which showed the best invitro responses, were subjected to invivo analysis. Both extracts caused decrease in trypanosome parasitemia and prolongation of mean survival days of the rats to 14.67days as compared with 6.83days in the negative control group. The extracts displayed dose-dependent significant (p ≤ 0.05) antitrypanosomal activities when compared with the negative and positive controls. Conclusion: The chloroform and ethyl acetate extracts of Annona muricata showed a relatively higher antitrypanosomal activity over other solvent extracts used in this study. Further fractionation, purification and isolation should be done to confirm the active components in this plant that is responsible for the antitrypanosomal activities recorded.
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番荔枝叶提取物体外和体内抗布氏锥虫活性研究
目的:锥虫病是一种影响家畜、野生动物和人类的重要原虫病。它是由属于锥虫属的采采蝇传播的细胞外血鞭毛虫引起的。在东非和南部非洲,非洲人类锥虫病是由布氏罗得西亚锥虫引起的,而在西非和中非,是由冈比亚锥虫引起的。另一方面,动物锥虫病是由布氏锥虫、间日锥虫和刚果锥虫引起的。在撒哈拉以南非洲,大约有6000万人面临感染的风险。本研究评价了番荔枝叶提取物对布氏锥虫感染的白化大鼠的抗锥虫作用。研究地点和时间:2021年2月至6月,尼日利亚阿库雷联邦科技大学动物生产与卫生系动物寄生虫学和微生物学研究单位。方法:采用不同的溶剂提取物(乙醇、乙酸乙酯、正己烷、氯仿和水),以2.5mg/ml、5mg/ml和10mg/ml的不同浓度进行体外抗锥虫体分析。乙酸地咪那嗪和生理盐水分别作为阳性对照和阴性对照。体内实验通过连续3天腹腔注射分级剂量(200、400和600mg/kg)的乙酸乙酯和氯仿提取物进行。结果:正己烷、氯仿和乙酸乙酯提取物对DPPH自由基的清除率分别为85.52%、80.00和78.49%。在体外试验中,各萃取物在不同时间均表现出活力下降的趋势。这些反应与浓度呈正相关。氯仿浓度为10mg/ml,乙酸乙酯浓度为10mg/ml,分别在35分钟和45分钟时使寄生虫完全停止运动。对体外反应最好的两种提取物(乙酸乙酯和氯仿提取物)进行了体内分析。与阴性对照组的6.83天相比,两种提取物均能降低大鼠的锥虫寄生率,使大鼠的平均生存天数延长至14.67天。与阴性对照和阳性对照相比,提取物的抗锥虫活性呈剂量依赖性显著(p≤0.05)。结论:三氯甲烷和乙酸乙酯提取物比其他溶剂提取物具有较高的抗锥虫活性。进一步的分离、纯化和分离,以确认在该植物负责抗锥虫活性记录的有效成分。
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