Phytochemical and antimicrobial investigations on various parts of Sida acuta...

[Full title: Phytochemical and antimicrobial investigations on various parts of Sida acuta Burm. f.]

71 Journal of Ayurvedic and Herbal Medicine 2018; 4(2): 71-75 Research Article ISSN: 2454-5023 J. Ayu. Herb. Med. 2018; 4(2): 71-75 © 2018, All rights reserved www.ayurvedjournal.com Received: 11-05-2018 Accepted: 03-07-2018 *Corresponding author: Chinelo A. Ezeabara Department of Botany, Nnamdi Azikiwe University, P.M.B. 5025 Awka, Nigeria Email: ca.ezeabara[at]unizik.edu.ng Phytochemical and antimicrobial investigations on various parts of Sida acuta Burm. f. Chinelo A. Ezeabara 1 *, Miracle O. Egenti 1 1 Department of Botany, Nnamdi Azikiwe University, P.M.B. 5025 Awka, Nigeria ABSTRACT Sida acuta Burm. f. belongs to the mallow family, Malvaceae. The study investigated the quantitative phytochemical contents and in vitro antimicrobial activities of S. acuta at different concentrations using standard techniques. Antimicrobial activity was evaluated by disc diffusion method and Minimum Inhibitory Concentrations for the absolute concentrations were determined by Agar well diffusion method. Test of signific ance was measured using Duncan’s Multiple Range Test. There were presence of alkaloid, flavonoid, saponin, tannin, cardiac glycosides, terpenoid, anthraquinone and steroid in the leaf, stem and root of the plant at varying levels. Alkaloid at 2.31±0.03 mg/100 g was the highest and it was detected in the leaf. All the plant parts exhibited inhibitory activities against all the test organisms but were dose-dependent. At 500 mg/ml, the leaf extract gave highest inhibition of Pseudomonas aeruginosa , Micrococcus varians , Candida albicans while the root extract gave highest inhibition of Escherichia coli , Salmonella typhi and Aspergillus flavus . Findings of this study, therefore, showed that all parts of S. acuta , particularly the leaf and the root, possessed antimicrobial properties which can be pharmaceutically harnessed. Keywords: Alkaloid, antibacteria, antifungi, Candida albicans , Escherichia coli , Aspergillus flavus . INTRODUCTION Sida acuta Burm. f. belongs to the genus Sida , of the mallow family, Malvaceae. It is commonly known as stubborn weed. It is an erect, branched, small perennial shrub with a woody tap root, and hairy branches up to 1 m high [1] . It is predominant in roadsides, waste areas, grazing land, disturbed land and abandoned farmlands. Traditional medicine has started gaining credence over the last decade. Different parts as well as whole plants are used in folk medicine for treatment of different afflictions. A wide range of medicinal uses of S. acuta have been extensively documented. The use of S. acuta in treatment of asthma, renal inflammation, colds, fever, headache, ulcer and worm infections in regions around Central America has been reported [2] . The leaves are used for their diuretic, demulcent, anthelmintic and wound healing properties [3] . Moreover, it is used as a medicine in treatment of liver disorders, urinary disease, nervous disorder, blood disorder, biliary disease [4] . In Nigeria, the leaves of S. acuta are chewed for treating gonorrhea [5] . Any substance that kills or inhibits the growth of microorganisms with negligible side effects on the host is considered as an antimicrobial [8] . In addition, it could be natural or man-made. The most common fatal bacterial diseases are respiratory infections [7] . Fungal infection normally happens when the immune system is weak or the microbes are too much for it to handle. Earlier work in antimicrobial actions of this plant were based on aerial parts, mainly leaf [8-10] , hence, there is need to investigate the underground parts. The objectives of this study therefore, were to determine the quantitative phytochemical compositions of various parts of S. acuta as well as its antimicrobial properties. MATERIALS AND METHODS Collection of Sample Matured S. acuta was collected from an abandoned farm land in Nibo, Anambra State in the month of May. The plant specimen was identified by Dr. C.A. Ezeabara, of the Department of Botany, Nnamdi Azikiwe University, Awka, Anambra State, where the voucher specimen was deposited. Preparation of Samples for Analysis The fresh plant parts were washed with clean water and oven dried at a temperature 65 o C for 12 hours. The leaves, stems and roots of S. acuta were later cut into bits with knife and then oven-dried at a

J Ayu Herb Med ǀ Vol 4 Issue 2 ǀ April- June 2018 72 temperature of 70 o C for 12 hours to remove all moisture. The samples were grounded in a mortar with a pestle, and then in a blender (Omega, USA) into powdered form. Extraction of Plant Materials Methanol Extraction The ethanol extract of the plant was prepared by soaking the powdered sample of the leaf, stem and root in 100 ml of ethanol. The whole set up was left for 72 hours at room temperature and thereafter filtered using Whatman filter paper. The extract was then concentrated using rotary evaporator and allowed the solvent to evaporate. The concentrated extract was stored in an air tight container in a refrigerator at 20 o C until it is required for analysis. Quantitative phytochemical analysis Alkaloid and flavonoid determinations in the test samples were determined by the gravimetric and acid hydrolysis gravimetric method of Harborne [11] respectively. The method used was described by AOAC [12] was used to determine saponin content. Tannin determination was done with Folin-Denis spectrophotometric method of Pearson [13] . Determination of anthraquinone was done with the method described by Ezeabara and Okonkwo [14] . Cardiac glycoside was determined by alkaline picarate colorimeter method as outlined by Trease and Evans [15] . Microbial Analysis The pure cultures of the microorganisms were obtained from Department of Microbiology, IITA Ibadan, Oyo State. The bacteria isolates include gram positive and the gram negative bacteria which include Escherichia coli, Salmonella typhi, Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa and Micrococcus varians. The fungi were Aspergillus niger, Candida albicans, Fusarium solani and Rhizopus sp. Preparation of Media The various media used were prepared according to manufacturers’ instructions as indicated on the product label. The quantities required were measured using a weighing balance (in grams) into a conical flask and dissolved in the appropriate volume of water using a measuring cylinder. The media were properly mixed and sterilized by autoclaving at 121° C for 15 minutes at 760 mmHg. Isolation of the Test Organisms Using a wire loop, a colony of the test organisms were collected from pure cultures at soil Microbiology Unit, IITA Ibadan, Oyo State, Nigeria. Antimicrobial Test Procedures Preparation of Stock Solution Stock solutions of the methanolic extracts were prepared by weighing 2.0 g of each methanolic extracts using electronic weighing machine in the department. This was then dissolved completely in 2.0 ml of Dimethyl sulfoxide (DMSO) in sterile test tubes giving a stock solution concentration of 1000 mg/1 ml (that is, 1000 mg/ml or 1 g/ml) per extract. The stock solution was then labeled approximately and stored in room temperature till required for use. Determination of Inhibitory Activity Inhibitory activity of methanol extract of leaf, stem and root of S. acuta antibiotics (positive control) and Dimethyl sulfoxide (negative control) were determined using disc diffusion method. 5 mm discs were impregnated with concentration of 1000 mg/ml of the methanolic extract, 100 mg/ml of antibiotic (Streptomycin) and 1000 mg/ml of DMSO. After the disc had taken up the methanolic extractions, and the controls, the disks were removed, dried and placed on the media which the test microorganisms were freshly inoculated, then incubated at 37°C within 24 hours for bacteria and 25°C within 72 hours for fungi. Antimicrobial activity was determined after 24 hours (for bacteria) of incubation and 72 hours (for fungi) of incubation by measuring the zone of inhibition around each paper disc in millimeters (mm) [16] . Determination of Minimum Inhibitory Concentration Minimum inhibitory concentrations (MIC) for the absolute (stock) concentrations were determined by agar well diffusion method; 5- sterile plates were prepared, and nutrient broth poured into each of plates, and then allowed drying. With some Standardized inoculums (106 cfu/ml), a loop full of the different test organisms were inoculated and streaked onto each of the 5-plates when dried. Then, 10-holes were dug using an agar borer for the varying concentrations of methanolic extracts of root, leaf and stem. A marker was used to rule each plate to separate 5-holes each for the methanolic extracts. Varying concentrations of the different extracts were done by serial dilution method using DMSO as diluent. The absolute/stock concentrations of extracts used were 1000 mg/ml which is 100% respectively. Five test tubes per extract were prepared on test tube rack. The dilutions were for the following ranges: 50% (500 mg/ml), 25% (250 mg/ml), 12.5% (125 mg/ml) and 6.25% (62.5 mg/ml) in each of the test tubes. DMSO served as the negative control. A measured 50 ul volume of each dilution was added aseptically into the holes seeded with the test organisms in the nutrient agar plate using a syringe which measures in ul and allowed in the incubator for 35°C in 24 hours. The lowest concentration of methanolic extracts showing a clear zone of inhibition was considered as the MIC [17] . Determination of Minimum Bactericidal/Fungicidal Concentration The plates with the Minimum Inhibitory Concentrations (MIC) were further incubated for another 24 hours at 35°C to test which organism would grow on the zones of inhibition. Those plates after 24 hours’ organisms were completely killed and clear zones remained were referred to as bactericidal for bacteria and fungicidal for fungi [18] . Statistical analysis Analysis of Variance (ANOVA) using SPSS version 21 was employed in analyzing the data collected from the study. Test of significance was measured using Duncan’s multiple range test (DMRT). RESULTS The result of the quantitative phytochemical compositions of methanol extracts of the leaf, stem and root of S. acuta was shown in Table 1 . The table revealed that the leaf extract had the highest compositions of alkaloids, flavonoids and steroids being 0.55±0.02, 2.31±0.03 and 1.85±0.04 mg/100 g respectively. Highest content of saponins at 0.81±0.05 mg/100 g was detected in the stem extract, while highest values of tannins, anthraquinone and cardiac glycosides being 1.67±0.03, 1.91±0.08 and 1.48±0.05 mg/100 g were found in the root extract respectively. The leaf had the highest inhibition against Staphylococcus aureus , Salmonella typhi and Escherichia coli being 7.11±0.04, 6.89±0.02 and 7.63±0.04 mm respectively at 62.5 mg/ml. At 125 mg/ml, the leaf showed the highest inhibition against S. aureus , Salmonella typhi and E.

J Ayu Herb Med ǀ Vol 4 Issue 2 ǀ April- June 2018 73 coli being 8.19±0.05, 7.93±0.05 and 8.67±0.03 mm respectively. The leaf also exhibited the highest level of inhibition against S. aureus , S. typhi and P. aeruginosa being 9.40±0.21, 10.77±0.14 and 9.68±0.41 mm respectively, at 250 mg/ml. At 500 mg/ml, the leaf had highest inhibition against P. aeruginosa and M. varians being 13.34±1.82 and 13.06±1.42 mm respectively. The stem had highest inhibition against S. aureus and Bacillus cereus at 12.61±1.86 and 12.61±0.47 mm respectively. The root restrained the growth of Salmonella typhi and E. coli being 13.96±0.78 and 13.77±1.15 mm respectively. In comparison among the control and the three plant extracts, the control gave the highest inhibition of all the pathogens. There was a significant difference in the inhibitory activities of the leaf, stem and root extracts of S. acuta against all the bacterial pathogens assayed (p<0.05) (Table 2). For fungal pathogens, the leaf extract gave the highest inhibition of Aspergillus flavus and Candida albicans being 8.15±0.05 and 8.23±0.02 mm respectively; the stem extract gave the highest inhibition of Rhizopus sp. at 7.44±0.03 mm while the root extract gave highest inhibition of Fusarium solani being 6.58±0.04 mm at 62.5 mg/ml concentration. At 125 mg/ml concentration, the leaf extract gave the highest inhibition of A. flavus and C. albicans being 10.24±0.05 and 10.30±0.08 mm respectively, while the stem extract gave the highest inhibition of F. solani and Rhizopus sp. at 8.67±0.021 and 9.55±0.07 mm respectively. At 250 mg/ml concentration, the leaf extract gave highest inhibition of A. flavus being 12.95±0.92 mm and C. albicans being 13.18±0.22 mm, while the stem extract gave highest inhibition of F. solani and Rhizopus sp. being 12.16±0.33 and 10.56±1.01 mm respectively. At 500 mg/ml concentration, the leaf extract gave highest inhibition of C. albicans being 14.46±0.49 mm, while the root extract gave highest inhibition of A. flavus , F. solani mm and Rhizopus sp. at 14.82±0.22, 13.49±0.29 and 12.87±0.57 mm respectively. In comparison between the control and plant extracts, the control gave highest inhibition of all the pathogens. There was a significant difference in the inhibitory activities of the leaf, stem and root extracts of S. acuta against all the fungi pathogens assayed (p<0.05) (Table 3). DISCUSSION Sida acuta parts are loaded with bioactive compounds that have powerful health benefits. Alkaloids occurred highest in the leaf which is thought to be the major active component, with others being detected in the lesser extent. The therapeutic values of plants depend on their chemical components which produce specific pharmacological activities on the human and animal body. Hence, leaf of this plant could be regarded as rich source of alkaloid. There were considerable antibacterial and antifungal activities against all the selected bacteria and fungi in all the concentrations with maximum activity at 500 mg/ml. This indicated that plant extract hinder the growth of microorganisms at a higher level. The root extract gave the highest inhibition against E. coli and Salmonella typhi while the leaf caused the greatest growth retardation of P. aeruginosa and M. varians . This might be due to high values of cardiac glucosides, anthraquinone and tannin in the root It has been reported that tannin exhibit antimicrobial activity through a variety of mechanisms [19] . These implied that the methanol root extracts of S. acuta could be applied for formulation of novel antibacterial drugs targeted against E. coli and Salmonella typhi while the leaf extract could be used for manufacture of drugs against P. aeruginosa and M. varians . These properties could be attributable to the highest level of alkaloids present in the leaf. It was reported that alkaloids extracted from Sanguisorba officianalis had antimicrobial properties against P. aeruginosa and E . coli [20] . For fungal pathogens, the highest zone of inhibition was observed in A. flavus and C. albicans at concentrations of 62.5, 125, and 250 mg/ml; which was exhibited by the leaf extracts. At 500 mg/ml, the leaf extract also gave the highest inhibition of C. albicans being 14.46±0.50 mm, while the root extract gave highest inhibition of A. flavus being 14.82±0.22 mm. This finding disagrees with the report of Oboh et al . [21] , who stated that ethanol extracts of S. acuta aerial parts had no inhibitory activity against E. coli, P. aeruginosa and C. albicans . These showed that A. flavus and C. albicans are susceptible to methanol extract of S. acuta . This might be due to high values of cardiac glucosides, anthraquinone and tannin in the root or either of the phytochemical This property is probably the reason behind the oral use of a decoction of the dried entire S. acuta plant for venereal diseases in Nicaragua [22] . This demonstrated that it can be used for synthesis of antifungal drugs for treatment of diseases caused by these organisms. In conclusion, considering the level of alkaloid in the leaf of this plant, it could be used as an analgesic. Moreover, the phytochemicals present in leaf, stem and root of S. acuta exhibited antibacterial and antifungal properties presenting it as a potent plant in treatment of bacterial and fungal infectious diseases. Hence, the bioactive agents could be isolated and incorporated in synthesis of new drugs. Table 1: Mean quantitative phytochemical compositions of the methanol extracts of Sida acuta leaf, stem and root Compositions (mg/100 g) Plant Parts Leaf Stem Root Alkaloid 2.31±0.03 c 0.96±0.04 a 0.34±0.00 b Cardiac glucosides 1.20±0.01 a 1.36±0.02 b 1.48±0.05 c Steroid Anthraquinone 1.85±0.04 c 1.62±0.02 b 1.66±0.03 b 1.50±0.04 a 0.70±0.05 a 1.48±0.05 c Saponin 0.28±0.05 a 0.81±0.05 b 0.26±0.01 a Tannin 1.51±0.02 b 1.34±0.01 a 1.67±0.03 c Flavonoid 0.55±0.02 b 0.23±0.01 a 0.25±0.07 a Results are in Mean± Std of triplicate determinations. Means with the same letter in a column is not significantly different (p>0.05) Table 2: Effects of methanol extracts of leaf, stem and root of Sida acuta on bacterial pathogens Concentrations (Mg/ml) Bacterial Strains Mean Zone of Inhibition (mm) ± SD Control Leaf Stem Root 62.5 Staphylococcus aureus 15.79±0.09 d 7.11±0.04 c 6.34±0.05 a 6.48±0.03 b Salmonella typhi 15.23±0.02 d 6.89±0.02 c 6.36±0.50 b 6.08±0.02 a Escherichia coli 15.52±0.010 d 7.63±0.04 c 7.21±0.06 b 6.87±0.05 a Pseudomonas aeruginosa 17.23±0.00 d 6.69±0.01 b 4.98±0.00 a 6.73±0.09 c Bacillus cereus 16.12±0.06 d 5.43±0.09 c 4.66±0.54 a 5.25±0.07 b Micrococcus varians 18.27±0.06 d 7.32±0.01 b 5.64±0.02 a 7.33±0.04 c

J Ayu Herb Med ǀ Vol 4 Issue 2 ǀ April- June 2018 74 125 Staphylococcus aureus 15.79±0.09 d 8.19±0.05 c 6.40±0.06 a 7.52±0.03 b Salmonella typhi 15.23±0.02 d 7.93±0.05 c 6.08±0.11 a 7.12±0.05 b Escherichia coli 15.52±0.01 d 8.67±0.03 c 6.27±0.06 a 7.89±0.05 b Pseudomonas aeruginosa 17.23±0.01 d 7.72±0.02 b 6.01±0.01 a 7.79±0.06 c Bacillus cereus 16.12±0.06 d 6.49±0.06 c 5.06±0.04 a 6.29±0.09 b Micrococcus varians 18.27±0.06 c 8.39±0.04 b 6.68±0.05 a 8.35±0.03 b 250 Staphylococcus aureus 15.79±0.09 c 9.40±0.21 b 8.57±0.17 a 9.19±0.60 b Salmonella typhi 15.23±0.02 d 10.77±0.14 c 8.01±0.45 a 10.00±0.22 b Escherichia coli 15.52±0.01 c 11.12±1.04 b 9.44±0.34 a 11.49±0.00 b Pseudomonas aeruginosa 17.23±0.00 c 9.68±0.41 b 8.85±0.59 a 8.44±0.30 a Bacillus cereus 16.12±0.06 d 7.47±0.36 b 6.42±0.26 a 8.04±0.30 c Micrococcus varians 18.27±0.06 c 10.28±0.61 b 9.38±0.28 a 10.35±0.39 b 500 Staphylococcus aureus 15.79±0.09 b 11.70±1.38 a 12.61±1.86 a 11.16±1.31 a Salmonella typhi 15.23±0.02 d 12.91±0.77 b 10.06±0.15 a 13.96±0.78 c Escherichia coli 15.52±0.01 c 11.66±0.70 a 10.72±0.59 a 13.77±1.15 b Pseudomonas aeruginosa 17.23±0.00 c 13.34±1.82 b 11.07±1.25 a 11.59±0.29 a Bacillus cereus 16.12±0.05 c 11.59±0.40 a 12.61±0.47 b 10.61±1.14 a Micrococcus varians 18.27±0.06 b 13.06±1.42 a 11.67±1.50 a 12.16±0.08 a Results are in Mean± Std of triplicate determinations. Means with the same letter in a column is not significantly different (p>0.05) Table 3: Effects of methanol extracts of leaf, stem and root of Sida acuta on fungal pathogens Concentrations( Mg/ml) Fungal Strains Mean Zone of Inhibition (mm) ± SD Control Leaf Stem Root 62.5 Aspergillus flavus 16.83±0.06 d 8.15±0.05 c 7.34±0.08 b 7.21±0.00 a Candida albicans 17.32±0.09 d 8.23±0.02 c 6.98±0.08 b 6.69±0.01 a Rhizopus sp 16.76±0.05 c 6.47±0.00 a 7.44±0.03 b 6.47±0.03 a Fusarium solani 17.02±0.05 b 6.56±0.06 a 6.55±0.00 a 6.58±0.04 a 125 Aspergillus flavus 16.83±0.06 d 10.24±0.05 c 9.46±0.09 b 9.26±0.03 a Candida albicans 17.32±0.09 d 10.30±0.08 c 9.09±0.07 b 8.75±0.06 a Rhizopus sp 16.76±0.05 c 8.58±0.02 a 9.55±0.07 b 8.55±0.05 a Fusarium solani 17.02±0.05 b 8.66±0.04 a 8.67±0.02 a 8.64±0.08 a 250 Aspergillus flavus 16.83±0.06 b 12.95±0.92 a 11.71±0.74 a 11.30±1.23 a Candida albicans 17.32±0.09 b 13.18±0.22 a 12.92±0.09 a 13.07±0.32 a Rhizopus sp 16.76±0.05 b 10.06±0.07 a 10.56±1.01 a 10.54±0.67 a Fusarium solani 17.02±0.05 c 11.07±0.91 a 12.16±0.33 b 10.98±0.25 a 500 Aspergillus flavus 16.83±0.06 d 13.29±0.26 b 12.70±0.13 a 14.82±0.22 c Candida albicans 17.32±0.09 d 14.46±0.49 c 12.23±0.30 b 11.38±0.10 a Rhizopus sp 16.76±0.05 c 12.75±0.73 b 11.76±0.46 a 12.87±0.57 b Fusarium solani 17.02±0.05 c 13.18±0.24 b 12.75±0.34 a 13.49±0.29 b Results are in Mean± Std of three different determinations. The same letter in a column is not significantly different (p>0.05). REFERENCES 1 Akobundu IO, Agyakwa CW. A Hand Book of West African Weeds . 2 nd ed. Ibadan: International Institute of Tropical Agriculture, 1998; 564 pp. 2 Caceres A, Giron LM, Martinez AM. Diuretic activity of plants used for treatment of urinary ailments in Guatemala. J. Ethnopharmacol 1987; 19:233-245. 3 Mohideen S, Sasikala E, Gopal V. Pharmacognostic studies on Sida acuta Burm. f. Anc. Sci. Life 2002; 22(1):57-66. 4 Sreedewi CD, Lartha PG, Ancy P, Suja SR, Shyamal S, Shine VJ, et al . Hepatoprotective studies on Sida acuta Burm. f. J. Ethnopharmacol 2009; 124(2):171-5. 5 Okafor JC. Tropical Plants in Health Care Delivery in Nigeria. Ibadan: Bookbuilders, 2013; 188 pp. 6 Aarestrup FM, Seyfarth AM, Emborg HD, Pedersen K, Henriksen RS, Bager F. Effect of abolishment of the use of antimicrobial agents for growth promotion on occurrence of antimicrobial resisitant in facal enterococci from food animals in Denmark. Antimicrobial Agents and Chemotherapy 2001; 45(7):2054-2059. 7 Sears CL. A dynamic partnership: Celebrating our gut flora. Anaerobe 2005; 11(5):247-51. 8 Ekpo MA, Etim PC. Antimicrobial activity of ethanol and aqueous extracts of Sida acuta on microorganisms from skin infections. Journal of Medicinal Plants Research 2009; 3(9):621-624. 9 Akilandeswari S, Senthamarai Prema S, Valarmathi R. Antimicrobial activity of leaf extracts of Sida acuta Burm. f. International Journal Pharma Sciences and Research 2010; 1(5):248-250. 10 Gopal PR, Salunke KJ. Evaluation of Sida acuta extract for antibacterial activity. International Journal of Plant Sciences 2013; 8(2):423-425. 11 Harborne JB. Phytochemical Methods. London: Chapman and Hall, 1973; 273 pp. 12 Association of Official Analytical Chemists, Official Methods of Analysis.17 th ed . Washington D.C.: International Association of Official Analytical Chemists, 2000; 2200 pp.

J Ayu Herb Med ǀ Vol 4 Issue 2 ǀ April- June 2018 75 13 Pearson D. Laboratory Techniques in Food Analysis. London: Butterworth, 1976. 14 Ezeabara CA, Okonkwo EE. Comparison of phytochemical and proximate components of leaf, stem and root of C roton hirtus L’Herit and Croton lobatus Linn. Journal of Pharma Science 2016; 1(3):47-56. 15 Trease GE, Evans WE. Pharmacognosy. 13 th ed. London: Bailhere Tindal, 1989; 832 pp 16 Reynolds D. Recruitment of Through 319-phosphorylated Ndd 1 p to the FHA domain of Fkh 2 p requires CLB kinase activity: a mechanism for CLB cluster gene activation. Genes Development 2003; 17(14):1789-802 17 Thongson C, Davidson PM, Mahakarnchanakul W, Weiss J. Antimicrobial activity of ultrasound-assisted solvent-extracted spices. Letters in Applied Microbiology 2004; 39:401-406. 18 Espinel-Ingroff A. E-Test Method for Testing Susceptibilities of Aspergillus spp. to the New Triazoles Voriconazole and Posaconazole and to Established Antifungal Agents: Comparison with NCCLS Broth Microdilution Method. Nature 2002; 2101-2107. 19 Scalbert A. Antimicrobial properties of tannins. Phytochemistry 1991; 30(12):3875-3883 20 Oboh IE, Akerele JO, Obasuyi O. Antimicrobial activity of the ethanol extract of the aerial parts of Sida acuta Burm. f. (Malvaceae). Tropical Journal of Pharmaceutical Research 2007; 6(4):809-813. 21 Janovska D, Kubikova K, Kokoska L. Screening for antimicrobial activity of some medicinal plants species of traditional Chinese medicine. Czech J. Food Sci 2003; 21:107-110. 22 Henning A, Pongpan H, Abraham Z. Glimpses of Indian Ethno botany. New Delhi: Oxford & Publishing Co. 2011; 308-320 pp. HOW TO CITE THIS ARTICLE Ezeabara CA, Egenti MO. Phytochemical and antimicrobial investigations on various parts of Sida acuta Burm. f. J Ayu Herb Med 2018;4(2):71-75.

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Flavonoid, Venereal disease, Anthelmintic, Traditional medicine, Infectious disease, Result, Therapeutic value, Medicinal use, Statistical analysis, Antimicrobial activities, Plant material, Antimicrobial properties, Antimicrobial activity, Dimethyl sulfoxide, Folk medicine, Disc diffusion method, Zone of inhibition, Minimum inhibitory concentration, Bactericidal, Wound healing properties, Staphylococcus aureus, Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Perennial shrub, Collection of sample, Duncan's multiple range test, Diuretic, Antimicrobial, Gram positive bacteria, Gram negative bacteria, Agar well diffusion method, Positive control, Ethanol extract, Methanol extract, Tannin, Methanol extraction, Phytochemical content, Negative control, Fungal infection, Bioactive compound, Analysis of variance, Steroid, Nutrient broth, Antibacterial and antifungal activities, Antifungal properties, Salmonella Typhi, Analgesic, Microbial analysis, Plant Extract, Antibacterial drug, Saponin, Alkaloid, Serial dilution method, Leaf extract, Stock Solution, Terpenoid, Stem extract, Aspergillus flavus, Test organism, Bacterial pathogen, Preparation of stock solution, Cardiac glycoside, Bacillus cereus, Fungicidal, Department of Botany, Mean zone of inhibition, In vitro antimicrobial activities, Bacterial disease, Quantitative phytochemical analysis, Fusarium solani, Nnamdi Azikiwe University, Oral use, Bioactive agent, New Drug, Root extract, Aerial part, Antifungal drug, Pure culture, Respiratory infection, Blood disorder, Nervous disorder, Standard techniques, Anthraquinone, Preparation of sample, Abstract, Biliary disease, Antibacterial and antifungal, Liver Disorder, Inhibitory activities, Antimicrobial investigation, Fungal Pathogen, Urinary disease, Malvaceae.