Antifungal Activity of Ziziphus Mucronata and Erythrina Abyssinica Bark Crude Extracts on Cryptococcus Neofomans and Candida Albicans Species
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British Journal of Pharmaceutical Research 10(3): 1-11, 2016, Article no.BJPR.23843 ISSN: 2231-2919, NLM ID: 101631759 SCIENCEDOMAIN international www.sciencedomain.org Antifungal Activity of Ziziphus mucronata and Erythrina abyssinica Bark Crude Extracts on Cryptococcus neofomans and Candida albicans Species Tapiwa E. Manyarara 1,2*, J. Chifamba 1,2 and Felistas T. Tarugarira 2 1School of Pharmacy, College of Health Sciences, University of Zimbabwe, P.O.Box MP 167, Mount Pleasant, Harare, Zimbabwe. 2Department of Pharmaceutical Technology, School of Industrial Sciences and Technology, Harare Institute of Technology, P.O.Box BE 277, Ganges Road, Belvedere, Harare, Zimbabwe. Authors’ contributions This work was carried out in collaboration between all authors. Author TEM designed the study, wrote the protocol and wrote the first draft of the manuscript. Authors TEM, FTT and JC managed the literature searches, analyses of the study performed the spectroscopy analysis. Authors JC and FTT managed the experimental process and author FTT identified the species of plant. All authors read and approved the final manuscript. Article Information DOI: 10.9734/BJPR/2016/23843 Editor(s): (1) Rafik Karaman, Bioorganic Chemistry, College of Pharmacy, Al-Quds University, USA. Reviewers: (1) Natthanej Luplertlop, Mahidol University, Bangkok, Thailand. (2) Wagner Loyola, Brazilian Corporation of Agricultural Research, Brazil. Complete Peer review History: http://sciencedomain.org/review-history/13238 Received 24 th December 2015 th Original Research Article Accepted 20 January 2016 Published 9th February 2016 ABSTRACT Aims: The study aims at extraction, characterization and identification of bioactive compounds with antifungal properties from the plant species E. abyssinica and Z. mucronata against Cyptococcal neoformans and Candida albicans. Study Design: Phytochemical screening of crude plant extracts and determination of their minimum inhibitory concentration using Agar disc diffusion method. Place and Duration of Study: Pharmaceutical Technology Lab, Harare Institute of Technology, February 2014 and May 2015. Methodology: In this study crude bark extracts of Ziziphus mucronata and Erythrina abyssinica _____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected]; Manyarara et al.; BJPR, 10(3): 1-11, 2016; Article no.BJPR.23843 from Zimbabwe were test for antifungal activity using the disc diffusion method against Cryptococcus neoformans , Candida albicans with fluconazole as the positive control. Phytochemical screening tests on the crude extracts were carried out to identify phytoconstituents. Results: Effective minimum inhibitory concentrations against C. albicans were found to be 20% w/v and 10%w/v for Ziziphus mucronata and showed moderate growth at a 5% w/v concentration. Erythrina abyssinica had effective minimum inhibitory concentrations at 25% w/v and 12.5% w/v with moderate fungal growth observed at 6.25% w/v. The same concentration ranges for both crude extracts showed similar antifungal activity for C. neoformans . Both crude bark extracts tested positive for tannins, saponins, flavonoids and alkaloids which contribute to the antifungal activity of the plant species. Conclusion: Candida albicans and Cryptococcus neoformans are the most common fungal species that cause opportunistic infections to occur in HIV and AIDS. The effects produced by these plants have proven that they can be used to develop pharmaceutical agents alleviate the symptoms associated with these infections. The crude plant extracts were found to be active against Candida albicans and Cryptococcus neoformans . Both crude bark extracts tested positive for tannins, saponins, flavonoids and alkaloids which contribute to the antifungal activity of the plant species. Keywords: Ziziphus mucronata; Erythrina abyssinica; cryptoccocal meningitis; Candida albicans; anti- fungal . 1. INTRODUCTION including the skin, prostate and medullary cavity of the bones. Cryptococcal meningitis is an opportunistic central nervous system (CNS) infection caused It may also complicate organ transplantation, by Cryptococcus neoformans , a ubiquitous reticuloendothelial malignancy, corticosteroid encapsulated fungus. It often develops in treatment, or sarcoidosis. Common symptoms of immune-compromised people, especially AIDS pulmonary involvement include fever, general patients. It is the third most frequent opportunistic feeling of discomfort, dry cough, pain in the infection of the central nervous system in HIV- membrane surrounding the lungs, and rarely, infected individuals. In patients with HIV/AIDS, hemoptysis (blood in sputum). The presentation the yearly incidence rate is between 2 and 7 in cryptococcosis varies with the site of infection cases per 1,000 people. It is far more common in and the patient’s immune status [4,5]. HIV/AIDS patients in sub-Saharan Africa, where these patients have a mortality rate that is Globally, it has been estimated that estimated to be 50 to 70% [1–3]. approximately 957,900 cases of cryptococcal meningoencephalitis occur each year, resulting Cryptococcus neoformans , a type of yeast found in more than 600,000 deaths. The region with the worldwide can cause pulmonary and central highest number of estimated cases in 2006 was nervous system (CNS) infections that can sub-Saharan Africa (720,000 cases; range, potentially spread to other areas of the body. 144,000 to 1.3 million), followed by South and This infection is called cryptococcosis . HIV/AIDS Southeast Asia (120,000 cases; range, 24,000 to patients are especially vulnerable to developing 216,000) [1,2]. Although the incidence of the infection. If the infection spreads from the cryptococcal meningoencephalitis has declined lungs to the CNS (brain and spinal cord) of an in patients who have access to antiretroviral HIV patient, the condition is considered an AIDS- therapy (ART), cryptococcal disease remains a defining illness. This means the patient's leading cause of mortality in the developing world condition has progressed to AIDS [1–3]. where access to ART is limited. A low CD4 cell count is the main predictor of risk of cryptococcal Most infections develop after the yeast has been meningoencephalitis; the vast majority of cases inhaled into the lungs. The fungus strongly occur among AIDS patients with a CD4 count resists phagocytosis. This means the immune <100 cells/microlitre [6,7]. system cells have to work hard to engulf the organism. Cryptococcosis usually starts with a The disease epidemic is commonly managed by pulmonary (lung) infection, which then spreads to initiation of ART to boost CD4 cell count and the CNS. If left untreated, the infection may reduce risk of occurrence due to poor immune continue to spread to other organs in the body, response from the hosts’ body. Treatment or 2 Manyarara et al.; BJPR, 10(3): 1-11, 2016; Article no.BJPR.23843 prophylaxis of cryptoccocal meningitis cases may Ziziphus mucronata is a dense leafy small to also be employed by the use of amphotericin B medium-size tree, which is 2-5 m tall. This plant (Ambisome©) or fluconazole. Due to its excellent belongs to family Rhamnaceae and genus penetration into cerebrospinal fluid, fluconazole Ziziphus . Z. mucronata is found in tropical area is the agent of choice for the treatment of and along moist riverbanks. The Latin name cryptococcal meningitis and for prophylaxis of ‘Ziziphus' means thorny and ‘ mucronata' refers to cryptococcosis in AIDS patients. Although the pointed leaves of this species. The genus fluconazole is generally less effective than either Ziziphus is of some historical importance. It is ketoconazole or itraconazole against believed that Christ's crown was made from Z. nonmeningeal coccidiomycosis, it is the preferred spina-christi Willd., a species which closely treatment for coccidioidal meningitis [1,3,7]. resembles Z. mucronata but grows from central Africa northwards [9]. This plant is widely Candida albicans are a fungal species distributed in southern Africa. In Namibia, responsible for yeast infections of the vaginal Z. mucronata is widespread throughout Namibia, and oral mucosa especially as an opportunistic except in the Namib Desert, north-west of Etosha infection in diseases like HIV/AIDS and and a few areas in the southeast [11] recorded uncontrolled diabetes. The species will normally Z. mucronata to be the most widely distributed exist of the human skin but weakening immune tree in South Africa. This plant flowers between system or homeostatic control for the body may March and June producing small, round, red- lead to their overgrowth and subsequent infection brown berries with thin dry flesh which appears [8]. Oropharyngeal candidiasis develops in the all year round but is mostly common between mouth and throat and can be invasive leading to December and June [11,12]. Local names for ulceration of the upper gastrointestinal tract and Z. mucronata includes buffalo thorn (English), systemic infection from the candida species. Omukaru (Herero), (Damara/Nama), omukekete Vaginal candiasis is often associated with white (Kwanyama), mukalu (Silozi) and omusheshete discharge, irritation and burning sensation [8]. It and muchecheni in shona. occurs as a result in the imbalance of vaginal bacterial flora due to a weakened immune The leaves and fruits of Z. mucronata are a system, imbalance between estrogen and source of food for livestock, especially goats. The progesterone hormones