Evaluation of In-Vitro Anti-Tuberculosis Activity of Tetrapleura tetraptera Crude and Fractions on Multidrug Resistant Mycobacterium tuberculosis


The management, control and elimination of tuberculosis (TB) have been difficult with the advent of HIV and cases of multidrug resistant (MDR-TB) tuberculosis. The cases of multidrug resistance to rifampicin and isoniazid pose greater challenges on first line and second line drugs to eliminate TB. The study is aimed at establishing anti-tuberculosis activity of Tetrapleura tetraptera against Mycobacterium tuberculosis and MDR-TB and the phytochemical present. The leaves of Tetrapleura tetraptera were collected, weighed, dried and pulverized to powder. The pulverized leaves of Tetrapleura tetraptera were subjected to 70% methanol extraction and screened for phytochemical. The crude extract was further purified into fractions using silica gel and thin layer chromatography techniques. M. tuberculosis and MDR-TB were obtained from positive acid fast bacilli sputa of TB patients and confirmed using GeneXpert to differentiate genotypic drug susceptible M. tuberculosis and MDR-TB. The sputa were digested using sodium hydroxide-cysteine technique and cultured in Middlebrook 7H9. The crude extract and fractions were screened for anti-tuberculosis activity using tetrazolium microtitre plate assay. The results showed that Tetrapleura tetraptera crude had activities against M. tuberculosis at 7.4 ± 0 mg/ml and 27.5 ± 0 mg/ml for MDR-TB. One of the fractions inhibited the growth of M. tuberculosis at 0.24 ± 0 mg/ml and MDR-TB at 0.89 ± 0 mg/ml. The phytochemical screened includes tannins, alkaloids, saponins, flavonoids, phenols and resins. T. tetraptra possesses anti-tuberculosis potential at low concentration on MDR-TB and can be a lead compound in drug development for the treatment of tuberculosis and multidrug resistant tuberculosis.

Share and Cite:

Izebe, K. , Ibrahim, K. , Onaolapo, J. , Oladosu, P. , Ya’aba, Y. , Njoku, M. , Shehu, M. , Ezeunala, M. and Ibrahim, Y. (2020) Evaluation of In-Vitro Anti-Tuberculosis Activity of Tetrapleura tetraptera Crude and Fractions on Multidrug Resistant Mycobacterium tuberculosis. Journal of Tuberculosis Research, 8, 165-176. doi: 10.4236/jtr.2020.83015.

1. Introduction

Tuberculosis (TB) is caused by Mycobacterium tuberculosis, a bacterium that infects a one third of the world population with 2 billion people infected and 1.8 million deaths in 2016 [1] [2]. According to WHO [2] estimates that between year 2000 and 2020 nearly one billion people will become newly infected with, 200 million people will get sick and another 35 million will die from tuberculosis. The advent of HIV/AIDS pandemic in the 1980s, struck a blow to this optimism and there has been a global resurgence of TB [3] - [9]. MDR tuberculosis is among the most worrisome element of the pandemic of antibiotic resistance because TB patients fail treatments which have high risk of death [10] [11] [12]. In 2016, the WHO estimated that 4.1% of new cases and 20% of the previously treated cases of TB were MDR-TB [2]. There were also approximately 190,000 deaths from MDR TB. More than half of these patients were in India, China and the Russia Federation. Among patients with infectious (smear positive) pulmonary disease, MDR TB was seen in 35% of newly detected cases, and in a massive 76% of previously treated patients [12]. There are high burden drug resistant TB countries including India and South Africa. Today MDR TB spreads unchecked in most of the world. It is fueled by poverty at the individual and family levels, limiting access to effective treatment and at the regional and national levels where under resourced governments lacks the capacity to tackle this disease [13]. After the introduction of rifampicin, no worthwhile antituberculosis drug with new mechanisms of action has been developed in over thirty years [11]. Globally, there is a challenge to develop new drugs especially for the MDR TB that is ravaging the pandemic of antibiotic resistance because TB patients fail treatments which have a high risk of death [4]. The available TB drugs used especially for the MDR TB have varied degrees of toxicity on the users. The plants are diverse in nature and source of future drugs lead [14] [15]. Anochie et al. [15] reported that some Nigerian medicinal plants claimed by the traditional medicinal practitioners to cure TB were found not active against M. tuberculosis, the plants include Crinum glaucum, Treculis aficana, Erthrina mildaedi, Fucus thonningis and Xylopis aethiopica. Nigerian plants have been reported to possess antitubercular activities. Ethnobotanical surveys on Nigerian medicinal plants have been reportedly carried in the different Nigeria geo-political zones. Ibekwe et al. [16] reported the findings of the efficacy of some Nigerian plants used by traditional medicine practitioners in the management of tuberculosis. Others reported the potentials of Nigerian plants in the treatment of tuberculosis in the Western part of Nigeria [17], Middle belt areas of Nigeria [3], Adamawa State of Nigeria [18]. Some medicinal plants indigenous in Ethiopia including Pterolobium stellatum, Vernonia amygdalina, Allium ursium and Dodonaea angulstifolia have been reported to possess anti-mycobacterial activity [13] [19]. Tetrapleura tetraptera is a woody plant growing in the deciduous and fringing forest. The plant can grow up to 80 ft high and over 4 ft girth with sharp buttresses. The foliage is dark and bi-pinnate leaves with thin bark. The leaves of T. tetraptera are commonly used as flavor because of the caramel-like smell [20]. The medicinal potentials have been explored by the locals in West Africa. The Binis use the seeds to improve taste of soup. The fruits are used as medicinal for children in Yorubaland of Nigeria. The common names include Twi by the Ashanti in Ghana [20], Ighirehimi in Esan (Edo) [21]. It is commonly used in soups for nursing mothers to prevent post-partum contractions [22]. The soft parts of the fruit and bark are known to contain sugars, tannins, traces of saponin and amino acids [22]. Many works have been done on the plants of T. tetraptera [22] [23] [24] [25] [26] but little had been reported on the anti-mycobacterial activity of the leaves of T. tetraptera in the literature. The study is aimed to investigate the anti-mycobacterial activity and phytochemical screening of the leaves of T. tetraptera.

2. Materials and Methods

2.1. Collection and Authentication of the Plants

The T. tetraptera plant materials were collected from the wild in forest reserve in Ihievbe Town in Owan East of Edo State of Nigeria. The leaves and seeds were identified and authenticated at the Herbarium Unit of National Institute for Pharmaceutical Research and Development with a voucher number NIPRD/H/6808. The leaves were washed thoroughly in tap water and with distilled water and air dried at room temperature to constant weight. The plant materials were sorted out and dry milled into the powder form, sieved and kept safe until needed in the Department of Microbiology and Biotechnology, National Institute for Pharmaceutical Research and Development.

2.2. Preparation of Extracts and Fractionation

Two hundred gram of the pulverized powder was extracted by maceration with 70% aqueous methanol for 72 hours at 25˚C on rotary shaker at 1000 rpm [27]. The extracts were then filtered and concentrated aseptically under reduced pressure using rotary evaporator to remove methanol and some water.

2.3. Preparation of Purified Fractions

The crude methanol extract of the plant was purified using column chromatography and thin layer chromatographer (TLC). Glass column was packed with silcal gel which then adsorbed the crude extract. It was then packed onto the column layer. The mobile phase consisted of three solvents, hexane, ethyl-acetate and methanol mixed in different proportions 100, 80, 60, 40, and 20 mL of methanol followed by 20, 40, 60, 80 and 100 mL ethyl-acetate. The various proportions of the solvents were pushed through the bed. Ten fractions were obtained. The fractions were pooled together by thin layer chromatography according to fingerprint (Table 1). This reduced the number of the fractions to six. Fractions A, B and C were used for anti-tubercular activity. The three fractions produced minute fraction and therefore not used in anti-mycobacterial assay.

2.4. Analysis for the Phytochemical Components

The plant extracts were screened for the presence of biological active components. The phyto-chemical tested for included alkaloids, anthraquinones, catachol, flavonoids, phenols, saponin, steroids, tri-terpenoids, and tannins using the standard methods. The standard techniques adopted were for screening the qualitative analysis of the phyto-chemical components. [14] [28].

2.5. Isolation and Identification of M. tuberculosis and MDR Tuberculosis Isolates from Patient Sputum

The sputa obtained from DOTS center of National Institute for Pharmaceutical Research and Development (NIPRD) were screened for acid fast bacilli (AFB) using Ziehl-Neelsen technique [29] [30] [31]. The positive AFB sputa were

Table 1. TLC spots on TLC plate of Tetrapleura tetraptera fractions.

analysed with GeneXpert [29] to confirm drug susceptible and multidrug-resistant tuberculosis. These sputa positive for drug susceptible and resistant TB were digested and decontaminated with NaOH-cysteine technique [32] [33] and were cultured in Middlebrook 7H9 (Sigma) for seven days with daily observation for turbidity and monitor for AFB with Ziehl-Neelsen technique.

2.6. Determination of Minimum Inhibitory Concentration

Two milligram each of the extract was dissolved in 1.0 mL dimethyl-sulphoxide (DMSO) to aid dissolution and made up with sterile distilled water to give 5.0 mL volume and filtered with syringe fixed membrane filters (0.45 µm). Two fold dilutions of the crude extract and fractions were made to obtain the desired starting concentrations mg per ml of the both crude and fractions to be used. 50 µl of sterile Middle Brook 7H9 broth was dispensed onto the 96 labeled micro well plates excluding the first row each of the eight rows. 100 µl of diluted extract was dispensed onto the first row of the 96 micro well plates. Two fold dilutions were carried out using 50 µl broth of each of the 1stwell and dispensed onto the 2nd well and continue to the 9th. The 10th well contained 100 µl of the extract concentration (extract sterility), 11th well contained 100 µl of media (media sterility control) and the 12th well contained the organism viability control. The procedure was repeated in triplicate. All inoculated plates were incubated at 37˚C for 5 days. At the end 5th day 25 µl of tetrazolium salt was added to wells to determine color change indicative of growth and not activity, while no color change showed further incubation for 48 hours. At end 48 hours 25 µl tetrazolium was further dispensed into controlled wells for activity by change in color. The absence of color change showed inhibition of M. tuberculosis (activity) and presence of color showed no inhibition or growth of M tuberculosis. The highest dilution that inhibits the growth of the Mycobacterium specie is the minimum inhibitory concentration of the plant extract. The above procedure was carried out for the different plants and on the different test Mycobacterium species. The above procedure was adopted from their works [34] [35].

2.7. Minimum Bactericidal Concentration

The results from the MIC plates were used for the MBC determination in the wells that did not show growth by means of no color change, 100 µl each was inoculated into sterile Middle Brook 7H10 agar and incubated at 37˚C for 7 days and count the number of colonies and compared with the initial growth per ml (105 cfu/ml). The minimum bactericidal concentration was computed if the growth was less than 0.1%. The procedure was adopted from his work [36].

2.8. Statistical Analysis

The statistical used included basic tool and graphical representation to analyze the data generated and use of SSPS version 22 software to analyze the data.

3. Results

3.1. Phytochemical Screening

Table 2 depicts the phytochemical composition of methanol extract of T. tetraptera. The qualitative test carried out on the T. tetraptera showed the presence of alkaloid, tannins, flavonoids and other phytochemical metabolites.

3.2. Anti-Tubercular Assay

Table 3 depicts the susceptibility profile of crude and fraction extracts of T. tetraptera against drug susceptible M. tuberculosis. The crude extract inhibited the growth of M. tuberculosis at the concentration of 7.4 ± 0 mg/ml (MIC) and 14.8 ± 0 mg/ml (MBC). The fractions A and B inhibited the growth of M. tuberculosis at concentrations of 0.25 ± 0 (MIC) and 0.50 ± 0 mg/ml (MBC). The fraction A extract had activity at concentrations of 0.11 ± 0 mg/ml (MIC) and 0.22 ± 0 mg/ml (MBC). The rifampicin had activity at 0.002 ± 0 mg/ml. The susceptible profile of M. tuberculosis as summarized in Figure 1.

Figure 1. MIC of crude and fractions of T. tetraptera and Rifampicin against M. tuberculosis.

Table 2. Phytochemical compositions of methanol crude extract T. tetraptera screened.

Table 4 depicts the susceptibility profile of crude and fraction extracts of T. tetraptera against MDR-TB isolates. The crude extract had activity against MDR-TB isolates at the concentration of 27.5 ± 0 mg/ml (MIC). The fraction C had activity against MDR-TB isolates at concentrations of 0.87 ± 0 mg/ml (MIC) and 1.78 ± 0 mg/ml (MBC). The fractions A and B extracts and rifampicin had no activity against MDR-TB isolates which was indicated with growths of MDR-TB isolates in the presence of different concentrations of fractions and rifampicin (0.002 mg). The susceptible pattern of T. tetraptera extracts and rifampicin summarized in Figure 2.

Figure 2. MBC of crude and fractions of T.tetraptera and rifampicin against M. tuberculosis.

Table 3. Anti-TB activity of crude extract and fractions T. tetraptera against drug susceptible M. tuberculosis.

Keys: (-) Activity, (+) No Activity, (A, B, C) fractions, (Rif) Rifampicin, MIC Minimum inhibitory concentration. MBC Minimum bactericidal concentration.

Table 4. Anti-TB activity of crude extract and fractions of T. tetraptera against multidrug-resistant tuberculosis.

Keys: (-) Activity, (+) No Activity, (R) Resistant (A, B, C) fractions, (Rif) Rifampicin ND Not Done, MIC Minimum inhibitory concentration. MBC Minimum bactericidal concentration.

4. Discussion

There have been ethno-botanic survey reports on the potentials of Nigerian medicinal plants used for the treatment of tuberculosis [15] [16] [17] [27]. Others reported on other plants from other countries for the management of TB in Ethiopian and South Africa [31] [37] [38] [39]. Akintola et al. [40] reported the anti-tubercular activity of Crinum jagus against M. tuberculosis. Similarly, Oladosu et al. [41] reported the activity of bioactive compounds from fruits extract of Acacia nilotica against M. tuberculosis. The information of activity of T. tetraptera on M. tuberculosis was found to be scanty and not documented.

The findings of this study revealed the presence of phytochemicals including tannins, alkaloids, saponins, flavonoids, phenols and resins with absence of terpenes, steroids, glycosides, and anthraquinones in T. tetraptra. The findings of this study agree the results of other researchers on phytochemical screening of T. tetraptera plant [23] [42] - [47]. The findings of this study also differed from report of Ebama [48] the presence of glycosides in the leaves of T. tetraptera. The presence of phytochemical secondary metabolites is responsible for the antimicrobial activity of the medicinal plants [49] [50]. Copp [51] reported presence of steroids, alkaloids; terpenes are associated with anti-mycobacterial activities. T. tetraptera methanol extracts contain saponnins, tannins, alkaloids, flavonoids. T. tetraptera does not have volatile oil and resins and may not have antioxidant potential. Abdel-Hamid et al. [39] reported the presence of glycosides, phenol, volatile oil, and resin that enhances immunity against pathology induced free radical generation.

T. tetraptera crude extract has anti-tubercular activity against susceptible M. tuberculosis at low concentration but differed with other researchers who reported lower concentrations of activity with different plants against M. tuberculosis [3] [16] [52] [53]. Adebiyi et al. [54] reported activity of Guiera senegalensis at 40 mg/ml against M. tuberculosis. Endale et al. [19] reported MICs range of 250 to 12.5 mg/ml for Allium ursinum and Dodonaea anguistfolia respectively. Kahaliw et al. [55] reported that Pterolobium stellatum had MIC of 0.039 mg/ml. The findings in this study, the MIC of T. tetraptera on M. tuberculosis was 7.4 mg/ml which was found lower in concentrations when compared with other reports. The methanol fractions of T. tetraptera showed much lower concentrations of 0.25 and 0.11 mg/ml (MICs) that inhibited the growth of M. tuberculosis. This study also revealed that the crude of T. tetraptera at the concentration of 27.5 mg/ml (MIC) had activity against MDR-tuberculosis isolates. The methanol fraction of T. tetraptera inhibited the growth of MDR-tuberculosis isolates at the concentration of 0.89 mg/ml (MIC), while the concentration of 0.002 mg/ml of rifampicin concentration was found to be resistant to MDR-TB. There was no information on the anti-tuberculosis activity of Tetrapleura tetraptera on M. tuberculosis and MDR-TB.

5. Conclusion

The T. tetraptera inhibited the growth of genetically analyzed drug susceptible M. tuberculosis at low concentrations. T. tetraptera is indicating a potential drug lead and the ability to further inhibition of MDR-TB that shows the diversity of the plant as it possesses anti-tuberculosis activity. T. tetraptera is a potent anti-tuberculosis drug against drug susceptible and MDR-TB.


We appreciate the Herbarium Unit of Department of Medicinal Plant Research and Traditional Medicine, National Institute for Pharmaceutical Research and Development for the identification of plant and extractions. We also wish to thank the Staff of Diagnostic Unit of Clinic staff for their contributions to the work.

Conflicts of Interest

The authors declare no conflicts of interest regarding the publication of this paper.


[1] World Health Organization (2017) Global Tuberculosis Report.
[2] World Health Organization (2018) Global Tuberculosis Report.
[3] Abdullahi, M., Ibrahim, K., Oyewale, A., Amupitan, J. and Okogun, J. (2009) Antimycobacterial Activity of some Medicinal Plants in Niger State. African Journal of Infectious Diseases, 3, 44-48.
[4] Dye, C., Espinal, M.A., Watt, J.C., Mbiaga, C. and Williams, B.G. (2002) Worldwide Incidence of Multidrug-Resistant Tuberculosis. The Journal of Infectious Diseases, 185, 1197-1202.
[5] Corbett, E.L., Churchyard, G.J., Charalambous, S., Samb, B., Moloi, V., Clayton, T.C., Grant, A.D., Murray, J., Hayes, R.J. and De Cork, K.M. (2003) Morbidity and Mortality in South African Gold Miners: Impart of Untreated Disease Due to Human Immunodeficiency Virus. Clinical Infectious Diseases, 34, 1251-1258.
[6] Mohan, A. and Sharma, S.K. (2001) Epidemiology. In: Sharma, S.K. and Mohan, A., Eds., Tuberculosis, Jayee Brothers Medical Publisher, New Delhi, 14-29.
[7] Raviglione, M.C., Snider Jr., D.E. and Kochi, A. (1995) Global Epidemiology of Tuberculosis: Morbidity and Mortality of a Worldwide Epidemic. JAMA, 273, 220-226.
[8] World Health Organization (2015) Global Tuberculosis Report.
[9] Orwa, C., Mutua, A., Kindt, R., Jamnadass, R. and Anthony, S. (2009) Agroforestry Database: A Tree Reference and Selection Guide Version 4.0, 1-5.
[10] Ormerod, I.P. (2005) Multidrug-Resistant Tuberculosis (MDR-TB): Epidemiology, Prevention and Treatment. British Medical Bulletin, 73-74, 17-24.
[11] Sharma, S.K. and Mohan, A. (2004) Multidrug-Resistant Tuberculosis. Indian Journal of Medical Research, 120, 354-367.
[12] Hoffner, S. (2012) Unexpected High Level of Multidrug-Resistant Tuberculosis Present New Challenges for Tuberculosis Control. The Lancet, 380, 1367-1369.
[13] Keshavjee, S. and Farmer, P.E. (2010) Time to Put Boots on the Ground Making Universe Access to MDR-TB Treatment a Reality. International Journal of Tuberculosis and Lung Disease, 14, 1222-1225
[14] Safowora, A. (1993) Medicinal Plant and Traditional Medicine in Africa. 2nd Edition, Spectrum Books Ltd., Ibadan Nigeria, 130.
[15] Anochie, P.I., Onyejepu, N., Ogu, A.C., Adentunji, M.A., Efere, L.O., Onnyezirila, A.C., Onyeneke, C.N., Obinna, J.U., Srikanth, A. and Bueno, J. (2011) Anti-Tuberculosis Activities of Medicinal Plants Used in the Treatment of Tuberculosis in HIV Patients in Nigeria. African Journal of Microbiology Research, 5, 1126-1130.
[16] Ibekw, N.N., Nvau, J.B., Oladosu, P.O., Usman, A.M., Ibrahim, K., Boshoff, H.I., Dowdc, C.S., et al. (2014) Some Nigerian Anti-Tuberculosis Ethnomedicines: A Preliminary Efficacy Assessment. Journal of Ethnopharmacology, 155, 524-532.
[17] Ogbole O. and Ajaiyeoba, O. (2010) Traditional Management of Tuberculosis in Ogun State of Nigeria: The Practice and Ethnobotanical Survey. African Journal of Traditional, Complementary and Alternative Medicines, 7, 79-84.
[18] Hassan, M.S., Kubmarawa, D., Oladosu, P. and Osemeahon, S.A. (2017) Evaluation of Some Medicinal Plants for Anti-Tuberculosis Activity from Adamawa State, Nigeria. Trends in Phytochemical Research, 1, 69-76.
[19] Endale, B., Berhan, M., Mebrahtom, G., Adane, W. and Gobena, A. (2014) Evaluation of In-Vitro Anti-Mycobacterial Activity of Selected Medicinal Plants in Mekelle, Ethiopia. World Applied Science Journal, 31, 1217-1220
[20] Irvine, F.R. (1961) Woody Plants of Ghana. Oxford University Press, Oxford, 341-343.
[21] Ugbogu, O.A., Soladoye, M.O., Baiyewu, R.A., and Ibhanesebhor, G.A. (2012) Vernacular Names of Some Nigerian Plants. Forestry Research, Benin City, 20.
[22] Adetola, O.O., Bodede, A.J. and Ogunbanjo, O.R. (2015) Nutritional and Medicinal Importance of Tetrapleura tetraptera Fruits (Aridan). Arican Journal of Science and Research, 4, 33-38.
[23] Adeleye, A.S., Oguoma, O.I., Braide, W. and Osogbase, E.E. (2016) Phytochemical and Antimicrobial Properties of 2:1 and 1:2 Ethanol-Methanol Extraction of Tetrapleura tetraptera. American Journal of current Microbiology, 4, 17-25.
[24] Abil, T.A. and Elegalam, A. (2007) Investigation into Chemical Composition of the Dry Fruits of Tetrapleura tetraptera (Ubukirhu). Journal of Food Technology, 5, 229-232.
[25] Borokini, T.I., Ighere, D.A., Clement, M., Ajiboye, T.O. and Aloowonle, A.A. (2013) Ethnoboiological Survey of Traditional Medicine Practices in Oyo State. Journal of Medicine Plant Studies, 1, 17-29.
[26] Okwu, J.A.O. (2001) Evaluation of the Chemical Composition of Indigenenous Spices and Flavouring Agents. Global Journal of Pure and Applied Science, 7, 445-459.
[27] Mann, A., Amupitan, J.O., Oyewale, A.O., Okogun, J.I., Ibrahim, K., Oladosu, P., Lawson, I., Olajide, I. and Nnamdi, A. (2008) Evaluation of in Vitro Antimycobacterial Activity of Nigerian Plants Used for the Treatment of Respiratory Diseases. African Journal of Biotechnology, 7, 1630-1636.
[28] Trease, G.E. and Evans, W. (2002) A Textbook of Pharmacognosy. 5th Edition, Elsevier Ltd., Edinburgh, 20-23.
[29] Centers for Disease Control and Prevention (2012) Trends in Tuberculosis-United States, 2012. MMWR, 62, 201.
[30] Ochei, J. and Kolhatkar, A. (2007) Medical Laboratory Science: Theory and Practice. 6th Edition, Tata McGraw-Hill, New York, 730.
[31] Cheesbourgh, M. (2002) Reaction Isolates on Tropical Disease. The Effects. Cambridge University Press, London, 76-100.
[32] Pfyffer, G.E., Brown-Elliot, B.A. and Wallace Jr., R.J. (2003) Mycobacterium. In: Murray, P.R., Baron, E.J., Jorgensen, J.H., Pfaller, M.A. and Yolken, R.H., Eds., Manual of Clinical Microbiology, 8th Edition, ASM Press, Washington DC, 532-584.
[33] Kent, P.T. and Kubica, G.P. (1985) Public Health Mycobacteriology: A Guide for the Level III Laboratory. US Department of Health and Human Services, Centers for Diseases Control, Atlanta, 36-38.
[34] Luz, C., Delgado, J. and Gilman, R.H. (2002) Tetrazolium Microplate Assay as a Rapid and Inexpensive Colorimetric Method for Determination of Antibiotic Susceptibility of Mycobacterium tuberculosis. Journal of Clinical Microbiology, 40, 873-1874.
[35] Ncube, N.S., Afolayan A.J. and Okoh, A.I. (2008) Assessment Techniques of Antimicrobial Properties of Natural Compounds of Plant Origin: Current Methods and Future Trends. African Journal of Biotechnology, 7, 1797-1806.
[36] Baron, E.J.O., Peterson, L.R. and Finegold, S.M. (1994) Bailey & Scott’s Diagnostic Microbiology. 9th Edition, Mosby Year Book, Maryland Heights, 168-175.
[37] Gizachew, Y.E., Giday, M. and Teklehaymanot, T. (2013) Antimycobacterial Activities of Selected Ethiopian Traditional Medicinal Plants Used for Treatment of Symptoms of Tuberculosis. Global Advanced Research Journal of Medicinal Plants (GARJMP), 2, 22-29.
[38] Semenya, S.S. and Maroyi, A. (2013) Medicinal Plants Used for the Tuberculosis by Bapedi Traditional Healers in Three Districts of Limpopo Province, South Africa. African Journal of Traditional, Complementary and Alternative, 10, 316-323.
[39] Abdel-Hamid, S.G., Bakhi, M.A., Yar, M.S., Al-Qasoumi, S.I. and Samad, A. (2010) Molecular Properties Prediction Synthesis and Antimicrobial Activity of Some Newer Oxadiazole Derivatives. European Journal of Medicinal Chemistry, 45, 5862-5869.
[40] Akintola, A.O., Kehinde, A.O., Adebiyi, O.E. and Ademowo, O.G. (2013) Anti-Tuberculosis of the Crude Methanolic Extract and Purified Fractions of the Bulb of Crinum jagus. Nigerian Journal of Physiological Sciences, 28, 135-140.
[41] Oladosu, P.O., Isu, N.R., Ibrahim, K., Orishadipe, A.T. and Lawson, L. (2013) Antituberculosis Activity of Bioactive Compounds from Fruit Extract of Acacia nilotica. Journal of Microbiology Research, 3, 247-254.
[42] Uchechi, E.N. and Chigozie, O.F. (2010) Antibacterial Activity of Tetrapleura tetraptera Taub. Pod Extracts. International Journal of Pharma and Bio Sciences, 1, 734-741.
[43] Akin-Idowu, P.E., Ibitoye, D.O., Ademoyegun, O.T. and Adeniyi, O.T. (2011) Chemical Composition of the Dry Fruit of Tetrapleura tetraptera and Its Potential Impact on Human Health. Journal of Herbs, Spices & Medicinal Plants, 17, 52-61.
[44] Mbota I.C., Dorenyi, O.J., Enyoga, I. and Udo Yoga, I (2013) Phytochemicals Properties and Antibacterial Activity of Tetrapleura tetraptera Fruit (Uyaka) on Medical Important Bacteria. Journal of Microbiology and Biotechnology, 3, 82-92.
[45] Koma, O.S., Olawumi, O.O., Godwin, E.-U. and Theophilus, O.A. (2016) Phytochemical Screening, In-Vitro Antimicrobial Activity and Antioxidant Characteristics of Tetrapleura tetraptera Extracts. European Journal of Plants, 18, 1-10.
[46] Gberikon, G.M., Agbulu, C.O. and Adeoti, I.I. (2016) Single and Combined Effects of Antibacterial Activities of Fruit and Stem Bark Extracts of Tetrapleura tetraptera on Streptococcus mutans and Pseudomonas aeruginosa. International Journal of Sciences, 4, 46-49.
[47] Ogugor, J.C., Ayuk, E.L , Osuji, A.S., Ugwu, M.O. and Unaegbu, M.E. (2018) Phytochemical, Antimicrobial, Proximate and Heavy Metals Analysis of Extracts of the Bark of Tetrapleura tetraptera Fruit. Global Scientific Journals, 6, 1-15.
[48] Ebama, R.U.B., Edet, U.D., Ekanemesang, G.M., Ikon, C.A., Etok, C.A. and Edet, A.P. (2016) Antimicrobial Activity, Phytochemical Screening and Nurient Analysis of Tetrapleura tetraptera and Piper guineense. Asian Journal of Medicine and Health, 1, 1-8.
[49] Rosa, M.D.S.S., Mendonca, F.R.R., Bizzo, H.R., Rodrigues, I.A., Soares, R.M.A., Souto-Padroin, T., Alviano, C.S. and Lopes, A.H.C.S. (2003) Antileishmanial Activity of a Linalool-Rich Essential Oil from Croton cajucara. Antimicrobial Agents and Chemotherapy, 47, 1895-1901.
[50] Vieira, R.H.S.F., Rodrigues, D.P., Goncalves, F.A., Menezee, F.G.R., Aragao, J.S., and Sousa, A.U. (2001) Microbial Effect of Medicinal Plant Extracts (Psdium guajava linn. and Carica papaya linn.) upon Bacteria Isolated from Fish Muscle and Known to Induce Diarrhea in Children. Revista Deinstitute de Medecina Tropica de Sao Paulo, 43, 145-148.
[51] Copp, B.R. (2003) Antimycobacterial Natural Products. Natural Product Reports, 20, 535-557.
[52] Oladosu, I.A., Ogundajo, A.L., Aiyelaagbe, O.O. and Emenyonu, N. (2011) Phytochemical and Anttuberculosis Activity of Coffea brivipes HIERN Extracts. Research Journal of Phytochemistry, 5, 130-135.
[53] Nvau, J.B., Oladosu, P.O., Orishadipe, A.T. (2011) Antimycobacterial Evaluation of Some Medicinal Plants Used in Plateau State of Nigeria for the Treatment of Tuberculosis. Agriculture and Biology Journal of North America, 2, 1270-1272.
[54] Adebayo, A., Ayo, R., Bello, I. and Habila, J. (2015) Phytochemical Screening and Anti-TB Activity of Root Extracts of Guirera senegalensis (J.F. Gmel). American Journal of Bioscience and Bioengineering, 3, 208-213.
[55] Kahaliw, W., Aseffa, A., Abebe, M., Teferi, M. and Engidawork, E. (2017) Evaluation of the Antimycobacterial Activity of Crude Extracts and Solvent Fractions of Selected Ethiopian Medicinal Plants. BMC Complementary and Alternative Medicine, 17, Article No. 143.

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.