Department of Microbial Biotechnology, Panjab University, Chandigarh, 160014, India.
School of Biotechnology, Jawaharlal Nehru University, New Delhi, 110067, India.
J Ethnopharmacol. 2021 Jan 10;264:113230. doi: 10.1016/j.jep.2020.113230. Epub 2020 Aug 24.
Gastrointestinal anthrax, a disease caused by Bacillus anthracis, remains an important but relatively neglected endemic disease of animals and humans in remote areas of the Indian subcontinent and some parts of Africa. Its initial symptoms include diarrhea and stomachache. In the current study, several common plants indicated for diarrhea, dysentery, stomachache or as stomachic as per traditional knowledge in the Indian subcontinent, i.e., Aegle marmelos (L.) Correa (Bael), Allium cepa L. (Onion), Allium sativum L. (Garlic), Azadirachta indica A. Juss. (Neem), Berberis asiatica Roxb. ex DC. (Daruharidra), Coriandrum sativum L. (Coriander), Curcuma longa L. (Turmeric), Cynodon dactylon (L.) Pers. (Bermuda grass), Mangifera indica L. (Mango), Morus indica L. (Black mulberry), Ocimum tenuiflorum L. (Ocimum sanctum L., Holy Basil), Ocimum gratissimum L. (Ram Tulsi), Psidium guajava L. (Guava), Zingiber officinale Roscoe (Ginger), were evaluated for their anti-Bacillus anthracis property. The usage of Azadirachta indica A. Juss. and Curcuma longa L. by Santals (India), and Allium sp. by biblical people to alleviate anthrax-like symptoms is well documented, but the usage of other plants is traditionally only indicated for different gastrointestinal disturbances/conditions.
Evaluate the above listed commonly available edible plants from the Indian subcontinent that are used in the traditional medicine to treat gastrointestinal diseases including those also indicated for anthrax-like symptoms for the presence of potent anti-B. anthracis activity in a form amenable to use by the general population in the endemic areas.
Aqueous extracts made from fourteen plants indicated above were screened for their anti-B. anthracis activity using agar-well diffusion assay (AWDA) and broth microdilution methods. The Aqueous Garlic Extract (AGE) that displayed most potent anti-B. anthracis activity was assessed for its thermostability, stability under pH extremes encountered in the gastrointestinal tract, and potential antagonistic interaction with bile salts as well as the FDA-approved antibiotics used for anthrax control. The bioactive fractions from the AGE were isolated by TLC coupled bioautography followed by their characterization using GC-MS.
Garlic (Allium sativum L.) extract was identified as the most promising candidate with bactericidal activity against B. anthracis. It consistently inhibited the growth of B. anthracis in AWDA and decreased the viable colony-forming unit counts in liquid-broth cultures by 6-logs within 6-12 h. The AGE displayed acceptable thermostability (>80% anti-B. anthracis activity retained on incubation at 50 °C for 12 h) and stability in gastric pH range (2-8). It did not antagonize the activity of FDA-approved antibiotics used for anthrax control. GC-MS analysis of the TLC separated bioactive fractions of AGE indicated the presence of previously unreported constituents such as phthalic acid derivatives, acid esters, phenyl group-containing compounds, steroids etc. CONCLUSION: The Aqueous Garlic Extract (AGE) displayed potent anti-B. anthracis activity. It was better than that displayed by Azadirachta indica A. Juss. (Neem) and Mangifera indica L., while Curcuma longa L. (Turmeric) did not show any activity under the assay conditions used. Further work should be undertaken to explore the possible application of AGE in preventing anthrax incidences in endemic areas.
肠胃炭疽病是由炭疽杆菌引起的疾病,在印度次大陆和非洲部分地区偏远地区的动物和人类中仍然是一种重要但相对被忽视的地方病。其最初的症状包括腹泻和胃痛。在目前的研究中,几种在印度次大陆的传统知识中被指示用于治疗腹泻、痢疾、胃痛或作为胃药的常见植物,即番石榴(Aegle marmelos(L.)Correa)(印度醋栗)、洋葱(Allium cepa L.)、大蒜(Allium sativum L.)、印度楝树(Azadirachta indica A. Juss.)(Neem)、小檗(Berberis asiatica Roxb. ex DC.)(Daruharidra)、香菜(Coriandrum sativum L.)、姜黄(Curcuma longa L.)、百喜草(Cynodon dactylon(L.)Pers.)(百慕大草)、芒果(Mangifera indica L.)、黑桑(Morus indica L.)、圣罗勒(Ocimum tenuiflorum L.)(神圣罗勒,Holy Basil)、罗勒(Ocimum gratissimum L.)(Ram Tulsi)、番石榴(Psidium guajava L.)、生姜(Zingiber officinale Roscoe),都被评估了其抗炭疽杆菌的特性。桑塔尔人(印度)使用印度楝树和姜黄,圣经人民使用葱属植物来缓解炭疽样症状,这是有据可查的,但其他植物的使用传统上仅指示用于不同的胃肠道疾病/状况。
评估印度次大陆上常用的、可食用的上述植物,这些植物在传统医学中用于治疗胃肠道疾病,包括也指示用于炭疽样症状的植物,以评估其是否具有潜在的抗炭疽杆菌活性,使其适用于流行地区的一般人群使用。
使用琼脂孔扩散法(AWDA)和肉汤微量稀释法对上述十四种植物的水提取物进行抗炭疽杆菌活性筛选。显示出最有效抗炭疽杆菌活性的大蒜水提取物(AGE),评估了其热稳定性、在胃肠道中遇到的极端 pH 值下的稳定性,以及与胆盐的潜在拮抗作用,以及用于炭疽控制的 FDA 批准的抗生素。AGE 的生物活性馏分通过 TLC 结合生物自显影进行分离,然后使用 GC-MS 进行表征。
大蒜(Allium sativum L.)提取物被鉴定为最有前途的候选物,对炭疽杆菌具有杀菌活性。它在 AWDA 中一致地抑制了炭疽杆菌的生长,并在液体肉汤培养物中在 6-12 小时内将活菌形成单位计数减少了 6 个对数。AGE 显示出可接受的热稳定性(在 50°C 孵育 12 小时后保留>80%的抗炭疽杆菌活性)和在胃 pH 值范围内的稳定性(2-8)。它不会拮抗用于炭疽控制的 FDA 批准的抗生素的活性。AGE 的 TLC 分离生物活性馏分的 GC-MS 分析表明,存在以前未报道的成分,如邻苯二甲酸衍生物、酸酯、含苯基的化合物、甾体等。
大蒜水提取物(AGE)显示出有效的抗炭疽杆菌活性。它优于印度楝树(Neem)和芒果(Mangifera indica L.),而姜黄(Curcuma longa L.)在使用的测定条件下没有显示出任何活性。应进一步开展工作,探索 AGE 在预防流行地区炭疽病中的可能应用。