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Phytochemical screening and in vitro antimicrobial activity of various parts of Cleome ciliata Schum. & Thonn.

Phytochemical screening and in vitro antimicrobial activity of various parts of Cleome ciliata... Abstract Cleome ciliata Schum. & Thonn. of the family Cleomaceae, is an annual herb. Phytochemical screening was carried out and the in vitro antimicrobial activities of leaf, stem and root extracts of C. ciliata were assessed using standard techniques. Duncan’s multiple range test was used to assess the significance of the results. The highest concentration of flavonoids (1.83 ± 0.04 mg/100 g), phenols (0.65 ± 0.00 mg/100 g) and terpenoids (1.93 ± 0.05 mg/100 g) was found in the leaf, stem and root respectively. Extracts of all the plant parts exhibited inhibitory activity against all test bacterial and fungal pathogens that are dose-dependent while the antagonistic effects of the leaf and root extracts were higher. The leaf extract showed the highest inhibition of Klebsiella pneumoniae, Streptococcus pneumoniae, Staphylococcus aureus, Salmonella typhi, Pseudomonas aeruginosa, Rhizopus stolonifer and Fusarium oxysporum; the root extract showed the highest inhibitory activity against K. pneumoniae and Penicillium chrysogenum while the three extracts were least effective against Escherichia coli, Shigella sonnei and Aspergillus niger. The study demonstrated that C. ciliata contains phytochemicals with antimicrobial properties and hence, its potential in antibacterial and antifungal drug development. Cleome ciliata, phytochemicals, antimicrobial actions, zone of inhibition, Rhizopus stolonifer, Penicillium chrysogenum Introduction Cleome is a dicot genus of the family Cleomaceae. Cleome ciliata Schum. & Thonn. is an annual herb that has digitately compound leaves with leaflets lanceolate to obovate, mostly three, acute at their apices; the stems are not prickly; the fruit is between 2.5 – 6 cm long and the flowers are white, lilac or pink in color (Crocker, 2000). Microorganisms that form a parasitic association with other organisms are classified as pathogens. Some bacteria and fungi can be pathogenic while some are not. Pathogenic bacteria are the major cause of human death and disease while harmful fungi cause infections in healthy persons or immunosuppressed patients. The search for novel potent plants and plant components against pathogenic microorganisms has become increasingly important, as a result of the development of antibiotic resistance in microbes (Ahmad, Aqil, and Owais, 2006). These days, researchers are investigating a wide range of indigenous medicinal plants for antimicrobial active principles with little or no side effects (Chandra et al., 2017). Secondary metabolites are a group of phytochemicals that are by-products of plant metabolism, and are not involved in their vital metabolic pathways. The medicinal quality of plants depends on their chemical constituents that have physiological activity in human systems (Kumar et al., 2009). Antimicrobial investigation of plant extracts and phytochemicals is the basis for antimicrobial drug discovery (Cseke et al., 2006). In a previous study, the phytochemical properties of parts of C. ciliata were determined (Ezeabara and Nwafulugo, 2015), but there is a need to evaluate the biological actions of the phytochemicals present, hence this study. The aim and objective of this study therefore, was to examine the antimicrobial potential of phytochemicals present in parts of C. ciliata. Materials and methods Sources of materials The root, stem and leaf of mature C. ciliata at the flowering stage were collected in May, from the Eastern Farm of the National Root Crop Research Institute, Umudike, Abia State, Nigeria. The plant was identified by Dr C.A. Ezeabara, a Plant Taxonomist in the Department of Botany, Nnamdi Azikiwe University. The voucher specimens were deposited in the herbarium of the same department. Preparation of samples The fresh plant parts were cut into pieces with a knife and then oven-dried at a temperature of 65°C for 12 hours. The samples were ground in a mortar with a pestle, and then in a blender into powdered form. The powdered samples were stored in an air-tight container prior to analyses. Quantitative phytochemical analysis Alkaloids, anthraquinones, flavonoids, phenols, saponins, sterols, terpenoids and tannins were quantitatively determined in the leaf, stem and root extracts of C. ciliata by standard methods (Trease and Evans, 2002). Microbial analysis Preparation of microorganisms for the experiment The pure cultures of the microorganisms were obtained from the Department of Microbiology, Biotechnology and Nuclear Agriculture Research Institute of the Ghana Atomic Energy Commission, Accra, Ghana. The bacteria isolates included Gram-positive Staphylococcus aureus and Streptococcus pneumoniae, and Gram-negative Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi and Shigella sonnei. The fungi used for testing were Aspergillus niger, Fusarium oxysporum, Penicillium chrysogenum and Rhizopus stolonifer. They were collected based on their clinical and pharmacological importance. Isolation and characterization of bacteria and fungi A measured 100 ml of distilled water was poured in a sterile bottle. A soil sample of 1 g was weighed out and added to the bottle of distilled water. The bottle was tightly covered with a lid and thoroughly shaken. One ml of the solution was aseptically and serially diluted. A 1-ml aliquot of a six-fold dilution was plated using potato dextrose agar (PDA) and Nutrient Agar for fungi and bacteria respectively. The plates were gently rotated to distribute the inoculums evenly in the plates and left to solidify under laminar airflow. Each of the inoculated plates was incubated at 37°C for 48 hours. After good growth of the colonies, distinct colonies were sub-cultured with fresh PDA and Nutrient Agar respectively using the streak plate technique and incubated again at 37°C for 72 hours. Each fungal colony was confirmed as the correct species by examining it under a light microscope (DM3000, Germany) using an oil immersion objective after staining with lactophenol in cotton blue dye, while the bacterial colonies were Gram-stained before viewing. Pure isolates were placed on PDA/nutrient agar slants and stored at 4°C until needed. The isolates were characterized based on their cultural and biochemical characteristics. Antimicrobial susceptibility testing The agar well diffusion method was used to determine the sensitivity to the plant extract, as outlined by the International Commission on Microbiological Specifications for Foods (ICMSF, 1998). The pure strains of bacteria were first grown in nutritional broth for 18 hours before use. They were later sub-cultured in Mueller-Hinton agar (Oxoid Ltd). Thirty-eight grams (38 g) of the medium was suspended in 1 litre of distilled water. The medium was completely dissolved by heating it with constant stirring and then boiled for 1 minute. The mixture was autoclaved at 121°C for 15 minutes and cooled at room temperature. Cooled Mueller-Hinton agar was poured into sterile petri dishes on a level laboratory bench and a uniform depth of 4 mm was obtained. The petri dishes were then left to solidify at room temperature. The prepared Mueller-Hinton agar was checked to ensure that the final pH was 7.3 ± 1 at 25°C. The wells were then bored into the agar medium using a sterile 6 mm cork borer. The wells were then filled up with the solution of the extract without allowing it to spill on the surface of the medium. The plates were allowed to stand on the laboratory bench for 1 hour before incubating them at 37°C for 24 hours. The plates were later observed for the zones of inhibition. The degree of inhibition was presented in terms of diameter of the inhibition zone as measured with a transparent ruler. The effect of the extract on bacteria and fungi was compared with that of a standard antibiotic; ampicillin and fungabacter for bacteria and fungi respectively at a concentration of 100 mg/ml. Statistical analysis SPSS version 21 was used to analyse the data. All data were checked for normality and homogeneity using the Kolmogorov–Smirnov and Leven median tests, respectively. Analysis of variance (F-test) was then applied, and Duncan’s multiple range test (DMRT) was used to measure the significance of any differences. The data were presented as mean ± standard error and mean ± standard deviation of three determinations for phytochemical and antimicrobial investigations respectively. Results and discussion The phytochemical composition of the extracts is shown in Table 1. This indicates that C. ciliata contains a wide range of phytochemicals. High concentrations of alkaloids, flavonoids, saponins, terpenoids and tannins were found in the leaf, stem and root. Moreover, the alkaloid, flavonoid, saponin and tannin contents of the C. ciliata parts that were measured in this study were higher than the levels reported by Ezeabara and Nwafulugo (2015). These are the classes of plant secondary metabolites that are most commonly present in plants. The variation in the levels of the phytochemicals could be a result of a number of factors. They could be dependent on location, season, as well as method of extraction. Ezeabara and Egwuoba (2016) reported that the synthesis and accumulation of phytochemicals depend on species, age, season and climatic factors. Phytochemicals accumulate in plant organs such as fruits, flowers, leaves, stems, and roots. Alkaloids (Zulak et al., 2006) and flavonoids (Crozier, Jaganath, and Clifford, 2006) play a defensive role in the plant against herbivores and pathogens. Table 1. Mean quantitative phytochemical composition of the leaf, stem and root of Cleome ciliata Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Results are in mean ± SE of triplicate determinations. The same letter in a column means that the respective means were not significantly different (p > 0.05). Open in new tab Table 1. Mean quantitative phytochemical composition of the leaf, stem and root of Cleome ciliata Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Results are in mean ± SE of triplicate determinations. The same letter in a column means that the respective means were not significantly different (p > 0.05). Open in new tab The high levels of these compounds in this study are most likely due to the solvent used in the extraction. In the earlier study (Ezeabara and Nwafulugo, 2015), ethanol was used for alkaloid and saponin extraction while water was used for tannin extraction. The higher concentrations in the current study suggest that methanol is a better solvent for the extraction of these phytochemicals and probably the major factor contributing to the detection of high levels of these biochemicals in C. ciliata. In the antimicrobial tests of the leaf, stem and root extracts of C. ciliata, the plant exhibited inhibitory activity against all test bacterial and fungal pathogens (Tables 2 and 3 and Figs 1–6). This revealed that compounds with antibacterial and antifungal actions are present in C. ciliata. Moreover, the result showed that the leaf extract had the highest inhibitory activity against K. pneumoniae, S. pneumoniae and S. aureus. It also showed high inhibitory activity against P. aeruginosa and S. typhi.Ezeabara and Egenti (2018) reported that a methanol leaf extract of Sida acuta Burm. f. at high concentration showed high inhibitory activity against P. aeruginosa (13.34 ± 1.82 (mm) ± SD). Pseudomonas is a common cause of urinary tract and wound infections where it causes acute or chronic infections (Ekhaise and Anyansi, 2005) while S. typhi is the causative agent for typhoid fever or enteric fever (Pavendan, Anbu, and Sebastian, 2011). The inhibitory activity against these microorganisms may be due to the presence of high levels of alkaloids, anthraquinones, flavonoids, saponins, sterols, terpenoids and tannins (1.41 ± 0.01, 0.74 ± 0.04, 1.83 ± 0.04, 1.52 ± 0.00, 0.50 ± 0.00, 1.73 ± 0.01 and 1.17 ± 0.01 mg/100 g respectively) in the leaf. Doss, Mubarack, and Dhanabalan (2009) reported that tannins isolated from Solanum trilobatum L. exhibited antibacterial activities against Escherichia coli, Pseudomonas aeruginosa, S. typhi and S. aureus. Moreover, some anthraquinones extracted from plants have been reported as therapeutic agents (Kanokmedhakul, Kanokmedhakul, and Phatchana, 2005) while flavonoids have a wide range of biological and pharmacological applications in the human system, probably as a result of their antimicrobial activities. Adedapo, Mogbojuri, Emikpe (2009) reported that saponins are of great pharmaceutical importance because of their relationship to compounds such as the sex hormones, steroids, vitamin D and cardiac glycosides. In addition, Sathya et al. (2012) reported that a terpenoid compound of Tridax procumbens showed antibacterial activity against both Gram-positive and Gram-negative bacteria. These findings imply that the leaf of C. ciliata will serve as a potential source of raw material in the pharmaceutical industry and in modern medicine for the treatment of human diseases caused by these bacterial pathogens. Table 2. Bacterial growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Results are in mean ± std of three replicates. Different letters in a column are significantly different (p < 0.05). Open in new tab Table 2. Bacterial growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Results are in mean ± std of three replicates. Different letters in a column are significantly different (p < 0.05). Open in new tab Table 3. Fungal growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Results are in mean ± std of three different determinations. The same letter in a column is not significantly different (p > 0.05). Open in new tab Table 3. Fungal growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Results are in mean ± std of three different determinations. The same letter in a column is not significantly different (p > 0.05). Open in new tab Figure 1. Open in new tabDownload slide Zone of Inhibition (mm) of bacterial pathogens by leaf extract of Cleome ciliata. Figure 2. Open in new tabDownload slide Zone of Inhibition (mm) of bacterial pathogens by stem extract of Cleome ciliata. Figure 3. Open in new tabDownload slide Zone of Inhibition (mm) of bacterial pathogens by root extract of Cleome ciliata. Figure 4. Open in new tabDownload slide Zone of Inhibition (mm) of fungal pathogens by leaf extract of Cleome ciliata. Figure 5. Open in new tabDownload slide Zone of Inhibition (mm) of fungal pathogens by stem extract of Cleome ciliata. Figure 6. Open in new tabDownload slide Zones of Inhibition (mm) of fungal pathogens by root extract of Cleome ciliata. The root extract of C. ciliata showed the highest inhibitory activity against K. pneumoniae and S. pneumoniae. This may be due to the high levels of alkaloids, saponins, terpenoids and tannins in the root. Organisms like S. pneumoniae have been associated with many human infections while K. pneumoniae has been found to account for up to 55% of nosocomial infections in some parts of Nigeria (Okonko et al., 2008). Moreover, the sensitivity and susceptibility of the test pathogens to the plant extracts was found to vary among the plant extracts. In particular, K. pneumoniae and S. pneumoniae were highly sensitive and susceptible to the plant extracts while Escherichia coli was the least susceptible. This suggested that E. coli is resistant to the inhibitory effects of the plant extract. Escherichia coli is associated with cholera, dysentery, gastroenteritis, typhoid fever and urinary tract infections (Orji, Ezenwaje, and Anyaegbunam, 2006). The methanol extracts of the leaf, stem and root of C. ciliata inhibited all the test fungi. Although the response of the test fungi varied among the plant extracts, the antagonistic effects of the leaf and root extracts were more evident than those of stem. This could be attributed to the higher concentration of bioactive compounds in the leaf and root than the stem. This implies that the leaf and root extracts of C. ciliata will serve as a better source of raw materials for the synthesis of new drugs that could be used to control and treat human and animal diseases, since A. niger, F. oxysporum, P. chrysogenum and R. stolonifer have all been associated with various diseases in humans and animals. Both the antibacterial and antifungal properties of the leaf, stem and root extracts of C. ciliata were found to increase with increasing concentration of the extract. This indicates that the antimicrobial actions of C. ciliata parts are dose dependent and that the three different parts have antimicrobial activities that can be fully utilized in phytomedicine. However, the standard antibiotics, ampicillin and fungabacter for bacteria and fungi, respectively, that were used as the standard control, showed higher inhibitory activities against all test bacteria and fungi. The superiority of conventional antibiotics over plant extracts has been extensively reported (Kubmarawa, Ajoku, and Okrie, 2002; Doss, Mubarack, and Dhanabalan, 2009; Samaila et al., 2014). This suggests that plant extracts need to be enhanced for better performance in health care delivery. Conclusion This study revealed that the methanol leaf extract contained the highest concentrations of anthraquinones and flavonoids while the root contained the highest concentrations of alkaloids, saponins, tannins and terpenoids. Moreover, they exhibited antibacterial and antifungal properties that were dose-dependent. The leaf extract showed the highest inhibition of K. pneumoniae, S. pneumoniae, S. aureus, S. typhi, P. aeruginosa, R. stolonifer and F. oxysporum while the root extract showed the highest inhibitory activity against K. pneumoniae and P. chrysogenum. Hence, these parts of C. ciliata will be very effective for the treatment of the diseases caused by these organisms. C. ciliata thus contains phytochemicals with antimicrobial properties, suggesting its potential use in the formulation of novel drugs. Author’s biography I.C.O. graduated with a Second Class Honours Upper Division in BSc Botany from Nnamdi Azikiwe University, Awka, Nigeria. His interest is in Phytomedicine. He performed the experiments. Dr C.A.E. designed and supervised the study and assisted in writing of the paper. References Adedapo , A. A. , Mogbojuri , O. M. and Emikpe , B. O. ( 2009 ) Safety evaluation of the aqueous extract of the leaves of Moringa oleifera , Journal of Medical Plants Research , 3 ( 8 ), 586 – 592 . Google Scholar OpenURL Placeholder Text WorldCat Ahmad , I. , Aqil , F. and Owais , M. ( 2006 ) Modern Phytomedicine: Turning Medicinal Plants into Drugs , Wiley-VCH , India , p. 404 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Chandra , H. , Bishnoi , P., Yadav , A. et al. . ( 2017 ) Antimicrobial resistance and the alternative resources with special emphasis on plant-based antimicrobials—a review , Plants , 6 ( 16 ), 11 . Google Scholar OpenURL Placeholder Text WorldCat Crocker , W. ( 2000 ) Growth of Plants , Reinhold Publishing Corporation, Ltd , London , p. 459 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Crozier , A. , Jaganath , I. B. and Clifford , M. N. ( 2006 ) Phenols, polyphenols and tannins: an overview, in Crozier A., Clifford M. N. and Ashihara H. (eds), Plant Secondary Metabolites: Occurrence, Structure and Role in the Human Diet , Blackwell Publishing Limited , Oxford, UK , pp. 1 – 24 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Cseke , L. J. , Kirakosyan , A., Kaufman , P. B. et al. . ( 2006 ) Natural Product from Plants , 2nd edn, CRC Press, Taylor & Francis Group, L.L.C , Boca Raton, Florida , p. 611 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Doss , A. , Mubarack , H. M. and Dhanabalan , R. ( 2009 ) Antibacterial activity of tannins from the leaves of Solanum trilobatum Linn , Indian Journal of Science and Technology , 2 ( 2 ), 41 – 43 . Google Scholar OpenURL Placeholder Text WorldCat Ekhaise , F. O. and Anyansi , C. C. ( 2005 ) Influence of brewery effluent discharge on the microbiological and physicochemical quality of Ikpoba River, Nigeria , African Journal of Biotechnology , 4 ( 10 ), 1062 – 1065 . Google Scholar OpenURL Placeholder Text WorldCat Ezeabara , C. A. and Egenti , M. O. ( 2018 ) Phytochemical and antimicrobial investigations on various parts of Sida acuta Burm. f , Journal of Ayurvedic and Herbal Medicine , 4 ( 2 ), 71 – 75 . Google Scholar OpenURL Placeholder Text WorldCat Ezeabara , C. A. and Egwuoba , G. C. ( 2016 ) Comparative screening of phytochemical and proximate constituents of leaf, stem and root of Oldenlandia corymbosa L. and Oldenlandia herbacea (L.) Roxb , American Journal of Life Science Research , 4 ( 3 ), 113 – 118 . Google Scholar OpenURL Placeholder Text WorldCat Ezeabara , C. A. and Nwafulugo , S. N. ( 2015 ) Comparison of phytochemical and proximate compositions of parts of Cleome ciliata Schum. & Thonn. and Cleome viscosa L. , World Journal of Biomedicine and Pharmaceutical Sciences , 1 ( 1 ), 1 – 5 . Google Scholar OpenURL Placeholder Text WorldCat International Commission on Microbiological Specifications for Foods . ( 1998 ) Principles for the establishment of microbiological food safety objectives and related control measures , Food Control , 9 ( 6 ), 379 – 384 . Crossref Search ADS WorldCat Kanokmedhakul , K. , Kanokmedhakul , S. and Phatchana , R. ( 2005 ) Biological activity of anthraquinones and triterpenoids from Prismatomeris fragrans , Journal of Ethnopharmacology , 100 ( 3 ), 284 – 288 . Google Scholar Crossref Search ADS PubMed WorldCat Kubmarawa , D. , Ajoku , G. and Okrie , D. A. ( 2002 ) Antimicrobial spectrum of hexane of Commiphora kerstingii Engl. (Buseraceae) , Technology Development , 8 , 29 – 32 . Google Scholar OpenURL Placeholder Text WorldCat Kumar , A. , Rajput , G., Dhatwalia , V. K. et al. . ( 2009 ) Phytocontent screening of Mucuna seeds and exploit in opposition to pathogenic microbes , Journal of Biological and Environmental Sciences , 3 , 71 – 76 . Google Scholar OpenURL Placeholder Text WorldCat Okonko , I. O. , Adejoye , O. D., Ogunnusi , T. A. et al. . ( 2008 ) Urban Domestic Solid Waste Management, Nimo , Rex Charles and Patrick Limited , Awka , pp. 5 – 7 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Orji , M. U. , Ezenwaje , E. E. and Anyaegbunam , B. C. ( 2006 ) Spatial appraisal of shallow well water pollution in Awka, Nigeria , Nigeria Journal of Microbiology , 20 ( 3 ), 1384 – 1389 . Google Scholar OpenURL Placeholder Text WorldCat Pavendan , P. , Anbu , S. S. and Sebastian , R. C. ( 2011 ) Physico Chemical and microbial assessment of drinking water from different water sources of Tiruchirappalli District, South India , European Journal of Experimental Biology , 1 ( 1 ), 183 – 189 . Google Scholar OpenURL Placeholder Text WorldCat Samaila , A. B. , Yarma , A. A., Oshomoh , E. O. et al. . ( 2014 ) Antimicrobial activity of the leaf and root extracts of Kigelia africana and Albizia chevalieri against Staphylococcus aureus , Greener Journal of Microbiology and Antimicrobials , 3 ( 1 ), 001 – 007 . Google Scholar Crossref Search ADS WorldCat Sathya , B. S. , Jayasurya , K. S., Sankaranarayanan , S., Bama , P. ( 2012 ) Antibacterial activity of different phytochemical extracts from the leaves of Tridax procumbens Linn.: identification and mode of action of the terpenoid compound as antibacterial , International Journal of Pharmacy and Pharmaceutical Sciences , 4 ( Suppl 1 ), 557 – 564 . Google Scholar OpenURL Placeholder Text WorldCat Trease , G. E. and Evans , W. C. ( 2002 ) Pharmacognosy , 15th edn, W.B. Saunders , London , p. 406 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Zulak , K. G. , Liscombe , D. K., Ashihara , H. et al. . ( 2006 ) Alkaloids, in Crozier A., Clifford M. N. and Ashihara H. (eds), Plant Secondary Metabolites: Occurrence, Structure and Role in the Human Diet , Blackwell Publishing Limited , Oxford, UK , pp. 102 – 136 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Author notes Supervisor: Dr Chinelo A. Ezeabara, Department of Botany, Nnamdi Azikiwe University, PMB 5025 Awka, Nigeria. © The Author(s) 2019. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com © The Author(s) 2019. Published by Oxford University Press. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png BioScience Horizons Oxford University Press

Phytochemical screening and in vitro antimicrobial activity of various parts of Cleome ciliata Schum. & Thonn.

Phytochemical screening and in vitro antimicrobial activity of various parts of Cleome ciliata Schum. & Thonn.

BioScience Horizons , Volume 11 – Jan 1, 2018

Abstract

Abstract Cleome ciliata Schum. & Thonn. of the family Cleomaceae, is an annual herb. Phytochemical screening was carried out and the in vitro antimicrobial activities of leaf, stem and root extracts of C. ciliata were assessed using standard techniques. Duncan’s multiple range test was used to assess the significance of the results. The highest concentration of flavonoids (1.83 ± 0.04 mg/100 g), phenols (0.65 ± 0.00 mg/100 g) and terpenoids (1.93 ± 0.05 mg/100 g) was found in the leaf, stem and root respectively. Extracts of all the plant parts exhibited inhibitory activity against all test bacterial and fungal pathogens that are dose-dependent while the antagonistic effects of the leaf and root extracts were higher. The leaf extract showed the highest inhibition of Klebsiella pneumoniae, Streptococcus pneumoniae, Staphylococcus aureus, Salmonella typhi, Pseudomonas aeruginosa, Rhizopus stolonifer and Fusarium oxysporum; the root extract showed the highest inhibitory activity against K. pneumoniae and Penicillium chrysogenum while the three extracts were least effective against Escherichia coli, Shigella sonnei and Aspergillus niger. The study demonstrated that C. ciliata contains phytochemicals with antimicrobial properties and hence, its potential in antibacterial and antifungal drug development. Cleome ciliata, phytochemicals, antimicrobial actions, zone of inhibition, Rhizopus stolonifer, Penicillium chrysogenum Introduction Cleome is a dicot genus of the family Cleomaceae. Cleome ciliata Schum. & Thonn. is an annual herb that has digitately compound leaves with leaflets lanceolate to obovate, mostly three, acute at their apices; the stems are not prickly; the fruit is between 2.5 – 6 cm long and the flowers are white, lilac or pink in color (Crocker, 2000). Microorganisms that form a parasitic association with other organisms are classified as pathogens. Some bacteria and fungi can be pathogenic while some are not. Pathogenic bacteria are the major cause of human death and disease while harmful fungi cause infections in healthy persons or immunosuppressed patients. The search for novel potent plants and plant components against pathogenic microorganisms has become increasingly important, as a result of the development of antibiotic resistance in microbes (Ahmad, Aqil, and Owais, 2006). These days, researchers are investigating a wide range of indigenous medicinal plants for antimicrobial active principles with little or no side effects (Chandra et al., 2017). Secondary metabolites are a group of phytochemicals that are by-products of plant metabolism, and are not involved in their vital metabolic pathways. The medicinal quality of plants depends on their chemical constituents that have physiological activity in human systems (Kumar et al., 2009). Antimicrobial investigation of plant extracts and phytochemicals is the basis for antimicrobial drug discovery (Cseke et al., 2006). In a previous study, the phytochemical properties of parts of C. ciliata were determined (Ezeabara and Nwafulugo, 2015), but there is a need to evaluate the biological actions of the phytochemicals present, hence this study. The aim and objective of this study therefore, was to examine the antimicrobial potential of phytochemicals present in parts of C. ciliata. Materials and methods Sources of materials The root, stem and leaf of mature C. ciliata at the flowering stage were collected in May, from the Eastern Farm of the National Root Crop Research Institute, Umudike, Abia State, Nigeria. The plant was identified by Dr C.A. Ezeabara, a Plant Taxonomist in the Department of Botany, Nnamdi Azikiwe University. The voucher specimens were deposited in the herbarium of the same department. Preparation of samples The fresh plant parts were cut into pieces with a knife and then oven-dried at a temperature of 65°C for 12 hours. The samples were ground in a mortar with a pestle, and then in a blender into powdered form. The powdered samples were stored in an air-tight container prior to analyses. Quantitative phytochemical analysis Alkaloids, anthraquinones, flavonoids, phenols, saponins, sterols, terpenoids and tannins were quantitatively determined in the leaf, stem and root extracts of C. ciliata by standard methods (Trease and Evans, 2002). Microbial analysis Preparation of microorganisms for the experiment The pure cultures of the microorganisms were obtained from the Department of Microbiology, Biotechnology and Nuclear Agriculture Research Institute of the Ghana Atomic Energy Commission, Accra, Ghana. The bacteria isolates included Gram-positive Staphylococcus aureus and Streptococcus pneumoniae, and Gram-negative Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi and Shigella sonnei. The fungi used for testing were Aspergillus niger, Fusarium oxysporum, Penicillium chrysogenum and Rhizopus stolonifer. They were collected based on their clinical and pharmacological importance. Isolation and characterization of bacteria and fungi A measured 100 ml of distilled water was poured in a sterile bottle. A soil sample of 1 g was weighed out and added to the bottle of distilled water. The bottle was tightly covered with a lid and thoroughly shaken. One ml of the solution was aseptically and serially diluted. A 1-ml aliquot of a six-fold dilution was plated using potato dextrose agar (PDA) and Nutrient Agar for fungi and bacteria respectively. The plates were gently rotated to distribute the inoculums evenly in the plates and left to solidify under laminar airflow. Each of the inoculated plates was incubated at 37°C for 48 hours. After good growth of the colonies, distinct colonies were sub-cultured with fresh PDA and Nutrient Agar respectively using the streak plate technique and incubated again at 37°C for 72 hours. Each fungal colony was confirmed as the correct species by examining it under a light microscope (DM3000, Germany) using an oil immersion objective after staining with lactophenol in cotton blue dye, while the bacterial colonies were Gram-stained before viewing. Pure isolates were placed on PDA/nutrient agar slants and stored at 4°C until needed. The isolates were characterized based on their cultural and biochemical characteristics. Antimicrobial susceptibility testing The agar well diffusion method was used to determine the sensitivity to the plant extract, as outlined by the International Commission on Microbiological Specifications for Foods (ICMSF, 1998). The pure strains of bacteria were first grown in nutritional broth for 18 hours before use. They were later sub-cultured in Mueller-Hinton agar (Oxoid Ltd). Thirty-eight grams (38 g) of the medium was suspended in 1 litre of distilled water. The medium was completely dissolved by heating it with constant stirring and then boiled for 1 minute. The mixture was autoclaved at 121°C for 15 minutes and cooled at room temperature. Cooled Mueller-Hinton agar was poured into sterile petri dishes on a level laboratory bench and a uniform depth of 4 mm was obtained. The petri dishes were then left to solidify at room temperature. The prepared Mueller-Hinton agar was checked to ensure that the final pH was 7.3 ± 1 at 25°C. The wells were then bored into the agar medium using a sterile 6 mm cork borer. The wells were then filled up with the solution of the extract without allowing it to spill on the surface of the medium. The plates were allowed to stand on the laboratory bench for 1 hour before incubating them at 37°C for 24 hours. The plates were later observed for the zones of inhibition. The degree of inhibition was presented in terms of diameter of the inhibition zone as measured with a transparent ruler. The effect of the extract on bacteria and fungi was compared with that of a standard antibiotic; ampicillin and fungabacter for bacteria and fungi respectively at a concentration of 100 mg/ml. Statistical analysis SPSS version 21 was used to analyse the data. All data were checked for normality and homogeneity using the Kolmogorov–Smirnov and Leven median tests, respectively. Analysis of variance (F-test) was then applied, and Duncan’s multiple range test (DMRT) was used to measure the significance of any differences. The data were presented as mean ± standard error and mean ± standard deviation of three determinations for phytochemical and antimicrobial investigations respectively. Results and discussion The phytochemical composition of the extracts is shown in Table 1. This indicates that C. ciliata contains a wide range of phytochemicals. High concentrations of alkaloids, flavonoids, saponins, terpenoids and tannins were found in the leaf, stem and root. Moreover, the alkaloid, flavonoid, saponin and tannin contents of the C. ciliata parts that were measured in this study were higher than the levels reported by Ezeabara and Nwafulugo (2015). These are the classes of plant secondary metabolites that are most commonly present in plants. The variation in the levels of the phytochemicals could be a result of a number of factors. They could be dependent on location, season, as well as method of extraction. Ezeabara and Egwuoba (2016) reported that the synthesis and accumulation of phytochemicals depend on species, age, season and climatic factors. Phytochemicals accumulate in plant organs such as fruits, flowers, leaves, stems, and roots. Alkaloids (Zulak et al., 2006) and flavonoids (Crozier, Jaganath, and Clifford, 2006) play a defensive role in the plant against herbivores and pathogens. Table 1. Mean quantitative phytochemical composition of the leaf, stem and root of Cleome ciliata Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Results are in mean ± SE of triplicate determinations. The same letter in a column means that the respective means were not significantly different (p > 0.05). Open in new tab Table 1. Mean quantitative phytochemical composition of the leaf, stem and root of Cleome ciliata Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Results are in mean ± SE of triplicate determinations. The same letter in a column means that the respective means were not significantly different (p > 0.05). Open in new tab The high levels of these compounds in this study are most likely due to the solvent used in the extraction. In the earlier study (Ezeabara and Nwafulugo, 2015), ethanol was used for alkaloid and saponin extraction while water was used for tannin extraction. The higher concentrations in the current study suggest that methanol is a better solvent for the extraction of these phytochemicals and probably the major factor contributing to the detection of high levels of these biochemicals in C. ciliata. In the antimicrobial tests of the leaf, stem and root extracts of C. ciliata, the plant exhibited inhibitory activity against all test bacterial and fungal pathogens (Tables 2 and 3 and Figs 1–6). This revealed that compounds with antibacterial and antifungal actions are present in C. ciliata. Moreover, the result showed that the leaf extract had the highest inhibitory activity against K. pneumoniae, S. pneumoniae and S. aureus. It also showed high inhibitory activity against P. aeruginosa and S. typhi.Ezeabara and Egenti (2018) reported that a methanol leaf extract of Sida acuta Burm. f. at high concentration showed high inhibitory activity against P. aeruginosa (13.34 ± 1.82 (mm) ± SD). Pseudomonas is a common cause of urinary tract and wound infections where it causes acute or chronic infections (Ekhaise and Anyansi, 2005) while S. typhi is the causative agent for typhoid fever or enteric fever (Pavendan, Anbu, and Sebastian, 2011). The inhibitory activity against these microorganisms may be due to the presence of high levels of alkaloids, anthraquinones, flavonoids, saponins, sterols, terpenoids and tannins (1.41 ± 0.01, 0.74 ± 0.04, 1.83 ± 0.04, 1.52 ± 0.00, 0.50 ± 0.00, 1.73 ± 0.01 and 1.17 ± 0.01 mg/100 g respectively) in the leaf. Doss, Mubarack, and Dhanabalan (2009) reported that tannins isolated from Solanum trilobatum L. exhibited antibacterial activities against Escherichia coli, Pseudomonas aeruginosa, S. typhi and S. aureus. Moreover, some anthraquinones extracted from plants have been reported as therapeutic agents (Kanokmedhakul, Kanokmedhakul, and Phatchana, 2005) while flavonoids have a wide range of biological and pharmacological applications in the human system, probably as a result of their antimicrobial activities. Adedapo, Mogbojuri, Emikpe (2009) reported that saponins are of great pharmaceutical importance because of their relationship to compounds such as the sex hormones, steroids, vitamin D and cardiac glycosides. In addition, Sathya et al. (2012) reported that a terpenoid compound of Tridax procumbens showed antibacterial activity against both Gram-positive and Gram-negative bacteria. These findings imply that the leaf of C. ciliata will serve as a potential source of raw material in the pharmaceutical industry and in modern medicine for the treatment of human diseases caused by these bacterial pathogens. Table 2. Bacterial growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Results are in mean ± std of three replicates. Different letters in a column are significantly different (p < 0.05). Open in new tab Table 2. Bacterial growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Results are in mean ± std of three replicates. Different letters in a column are significantly different (p < 0.05). Open in new tab Table 3. Fungal growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Results are in mean ± std of three different determinations. The same letter in a column is not significantly different (p > 0.05). Open in new tab Table 3. Fungal growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Results are in mean ± std of three different determinations. The same letter in a column is not significantly different (p > 0.05). Open in new tab Figure 1. Open in new tabDownload slide Zone of Inhibition (mm) of bacterial pathogens by leaf extract of Cleome ciliata. Figure 2. Open in new tabDownload slide Zone of Inhibition (mm) of bacterial pathogens by stem extract of Cleome ciliata. Figure 3. Open in new tabDownload slide Zone of Inhibition (mm) of bacterial pathogens by root extract of Cleome ciliata. Figure 4. Open in new tabDownload slide Zone of Inhibition (mm) of fungal pathogens by leaf extract of Cleome ciliata. Figure 5. Open in new tabDownload slide Zone of Inhibition (mm) of fungal pathogens by stem extract of Cleome ciliata. Figure 6. Open in new tabDownload slide Zones of Inhibition (mm) of fungal pathogens by root extract of Cleome ciliata. The root extract of C. ciliata showed the highest inhibitory activity against K. pneumoniae and S. pneumoniae. This may be due to the high levels of alkaloids, saponins, terpenoids and tannins in the root. Organisms like S. pneumoniae have been associated with many human infections while K. pneumoniae has been found to account for up to 55% of nosocomial infections in some parts of Nigeria (Okonko et al., 2008). Moreover, the sensitivity and susceptibility of the test pathogens to the plant extracts was found to vary among the plant extracts. In particular, K. pneumoniae and S. pneumoniae were highly sensitive and susceptible to the plant extracts while Escherichia coli was the least susceptible. This suggested that E. coli is resistant to the inhibitory effects of the plant extract. Escherichia coli is associated with cholera, dysentery, gastroenteritis, typhoid fever and urinary tract infections (Orji, Ezenwaje, and Anyaegbunam, 2006). The methanol extracts of the leaf, stem and root of C. ciliata inhibited all the test fungi. Although the response of the test fungi varied among the plant extracts, the antagonistic effects of the leaf and root extracts were more evident than those of stem. This could be attributed to the higher concentration of bioactive compounds in the leaf and root than the stem. This implies that the leaf and root extracts of C. ciliata will serve as a better source of raw materials for the synthesis of new drugs that could be used to control and treat human and animal diseases, since A. niger, F. oxysporum, P. chrysogenum and R. stolonifer have all been associated with various diseases in humans and animals. Both the antibacterial and antifungal properties of the leaf, stem and root extracts of C. ciliata were found to increase with increasing concentration of the extract. This indicates that the antimicrobial actions of C. ciliata parts are dose dependent and that the three different parts have antimicrobial activities that can be fully utilized in phytomedicine. However, the standard antibiotics, ampicillin and fungabacter for bacteria and fungi, respectively, that were used as the standard control, showed higher inhibitory activities against all test bacteria and fungi. The superiority of conventional antibiotics over plant extracts has been extensively reported (Kubmarawa, Ajoku, and Okrie, 2002; Doss, Mubarack, and Dhanabalan, 2009; Samaila et al., 2014). This suggests that plant extracts need to be enhanced for better performance in health care delivery. Conclusion This study revealed that the methanol leaf extract contained the highest concentrations of anthraquinones and flavonoids while the root contained the highest concentrations of alkaloids, saponins, tannins and terpenoids. Moreover, they exhibited antibacterial and antifungal properties that were dose-dependent. The leaf extract showed the highest inhibition of K. pneumoniae, S. pneumoniae, S. aureus, S. typhi, P. aeruginosa, R. stolonifer and F. oxysporum while the root extract showed the highest inhibitory activity against K. pneumoniae and P. chrysogenum. Hence, these parts of C. ciliata will be very effective for the treatment of the diseases caused by these organisms. C. ciliata thus contains phytochemicals with antimicrobial properties, suggesting its potential use in the formulation of novel drugs. Author’s biography I.C.O. graduated with a Second Class Honours Upper Division in BSc Botany from Nnamdi Azikiwe University, Awka, Nigeria. His interest is in Phytomedicine. He performed the experiments. Dr C.A.E. designed and supervised the study and assisted in writing of the paper. References Adedapo , A. A. , Mogbojuri , O. M. and Emikpe , B. O. ( 2009 ) Safety evaluation of the aqueous extract of the leaves of Moringa oleifera , Journal of Medical Plants Research , 3 ( 8 ), 586 – 592 . Google Scholar OpenURL Placeholder Text WorldCat Ahmad , I. , Aqil , F. and Owais , M. ( 2006 ) Modern Phytomedicine: Turning Medicinal Plants into Drugs , Wiley-VCH , India , p. 404 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Chandra , H. , Bishnoi , P., Yadav , A. et al. . ( 2017 ) Antimicrobial resistance and the alternative resources with special emphasis on plant-based antimicrobials—a review , Plants , 6 ( 16 ), 11 . Google Scholar OpenURL Placeholder Text WorldCat Crocker , W. ( 2000 ) Growth of Plants , Reinhold Publishing Corporation, Ltd , London , p. 459 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Crozier , A. , Jaganath , I. B. and Clifford , M. N. ( 2006 ) Phenols, polyphenols and tannins: an overview, in Crozier A., Clifford M. N. and Ashihara H. (eds), Plant Secondary Metabolites: Occurrence, Structure and Role in the Human Diet , Blackwell Publishing Limited , Oxford, UK , pp. 1 – 24 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Cseke , L. J. , Kirakosyan , A., Kaufman , P. B. et al. . ( 2006 ) Natural Product from Plants , 2nd edn, CRC Press, Taylor & Francis Group, L.L.C , Boca Raton, Florida , p. 611 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Doss , A. , Mubarack , H. M. and Dhanabalan , R. ( 2009 ) Antibacterial activity of tannins from the leaves of Solanum trilobatum Linn , Indian Journal of Science and Technology , 2 ( 2 ), 41 – 43 . Google Scholar OpenURL Placeholder Text WorldCat Ekhaise , F. O. and Anyansi , C. C. ( 2005 ) Influence of brewery effluent discharge on the microbiological and physicochemical quality of Ikpoba River, Nigeria , African Journal of Biotechnology , 4 ( 10 ), 1062 – 1065 . Google Scholar OpenURL Placeholder Text WorldCat Ezeabara , C. A. and Egenti , M. O. ( 2018 ) Phytochemical and antimicrobial investigations on various parts of Sida acuta Burm. f , Journal of Ayurvedic and Herbal Medicine , 4 ( 2 ), 71 – 75 . Google Scholar OpenURL Placeholder Text WorldCat Ezeabara , C. A. and Egwuoba , G. C. ( 2016 ) Comparative screening of phytochemical and proximate constituents of leaf, stem and root of Oldenlandia corymbosa L. and Oldenlandia herbacea (L.) Roxb , American Journal of Life Science Research , 4 ( 3 ), 113 – 118 . Google Scholar OpenURL Placeholder Text WorldCat Ezeabara , C. A. and Nwafulugo , S. N. ( 2015 ) Comparison of phytochemical and proximate compositions of parts of Cleome ciliata Schum. & Thonn. and Cleome viscosa L. , World Journal of Biomedicine and Pharmaceutical Sciences , 1 ( 1 ), 1 – 5 . Google Scholar OpenURL Placeholder Text WorldCat International Commission on Microbiological Specifications for Foods . ( 1998 ) Principles for the establishment of microbiological food safety objectives and related control measures , Food Control , 9 ( 6 ), 379 – 384 . Crossref Search ADS WorldCat Kanokmedhakul , K. , Kanokmedhakul , S. and Phatchana , R. ( 2005 ) Biological activity of anthraquinones and triterpenoids from Prismatomeris fragrans , Journal of Ethnopharmacology , 100 ( 3 ), 284 – 288 . Google Scholar Crossref Search ADS PubMed WorldCat Kubmarawa , D. , Ajoku , G. and Okrie , D. A. ( 2002 ) Antimicrobial spectrum of hexane of Commiphora kerstingii Engl. (Buseraceae) , Technology Development , 8 , 29 – 32 . Google Scholar OpenURL Placeholder Text WorldCat Kumar , A. , Rajput , G., Dhatwalia , V. K. et al. . ( 2009 ) Phytocontent screening of Mucuna seeds and exploit in opposition to pathogenic microbes , Journal of Biological and Environmental Sciences , 3 , 71 – 76 . Google Scholar OpenURL Placeholder Text WorldCat Okonko , I. O. , Adejoye , O. D., Ogunnusi , T. A. et al. . ( 2008 ) Urban Domestic Solid Waste Management, Nimo , Rex Charles and Patrick Limited , Awka , pp. 5 – 7 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Orji , M. U. , Ezenwaje , E. E. and Anyaegbunam , B. C. ( 2006 ) Spatial appraisal of shallow well water pollution in Awka, Nigeria , Nigeria Journal of Microbiology , 20 ( 3 ), 1384 – 1389 . Google Scholar OpenURL Placeholder Text WorldCat Pavendan , P. , Anbu , S. S. and Sebastian , R. C. ( 2011 ) Physico Chemical and microbial assessment of drinking water from different water sources of Tiruchirappalli District, South India , European Journal of Experimental Biology , 1 ( 1 ), 183 – 189 . Google Scholar OpenURL Placeholder Text WorldCat Samaila , A. B. , Yarma , A. A., Oshomoh , E. O. et al. . ( 2014 ) Antimicrobial activity of the leaf and root extracts of Kigelia africana and Albizia chevalieri against Staphylococcus aureus , Greener Journal of Microbiology and Antimicrobials , 3 ( 1 ), 001 – 007 . Google Scholar Crossref Search ADS WorldCat Sathya , B. S. , Jayasurya , K. S., Sankaranarayanan , S., Bama , P. ( 2012 ) Antibacterial activity of different phytochemical extracts from the leaves of Tridax procumbens Linn.: identification and mode of action of the terpenoid compound as antibacterial , International Journal of Pharmacy and Pharmaceutical Sciences , 4 ( Suppl 1 ), 557 – 564 . Google Scholar OpenURL Placeholder Text WorldCat Trease , G. E. and Evans , W. C. ( 2002 ) Pharmacognosy , 15th edn, W.B. Saunders , London , p. 406 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Zulak , K. G. , Liscombe , D. K., Ashihara , H. et al. . ( 2006 ) Alkaloids, in Crozier A., Clifford M. N. and Ashihara H. (eds), Plant Secondary Metabolites: Occurrence, Structure and Role in the Human Diet , Blackwell Publishing Limited , Oxford, UK , pp. 102 – 136 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Author notes Supervisor: Dr Chinelo A. Ezeabara, Department of Botany, Nnamdi Azikiwe University, PMB 5025 Awka, Nigeria. © The Author(s) 2019. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com © The Author(s) 2019. Published by Oxford University Press.

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Abstract

Abstract Cleome ciliata Schum. & Thonn. of the family Cleomaceae, is an annual herb. Phytochemical screening was carried out and the in vitro antimicrobial activities of leaf, stem and root extracts of C. ciliata were assessed using standard techniques. Duncan’s multiple range test was used to assess the significance of the results. The highest concentration of flavonoids (1.83 ± 0.04 mg/100 g), phenols (0.65 ± 0.00 mg/100 g) and terpenoids (1.93 ± 0.05 mg/100 g) was found in the leaf, stem and root respectively. Extracts of all the plant parts exhibited inhibitory activity against all test bacterial and fungal pathogens that are dose-dependent while the antagonistic effects of the leaf and root extracts were higher. The leaf extract showed the highest inhibition of Klebsiella pneumoniae, Streptococcus pneumoniae, Staphylococcus aureus, Salmonella typhi, Pseudomonas aeruginosa, Rhizopus stolonifer and Fusarium oxysporum; the root extract showed the highest inhibitory activity against K. pneumoniae and Penicillium chrysogenum while the three extracts were least effective against Escherichia coli, Shigella sonnei and Aspergillus niger. The study demonstrated that C. ciliata contains phytochemicals with antimicrobial properties and hence, its potential in antibacterial and antifungal drug development. Cleome ciliata, phytochemicals, antimicrobial actions, zone of inhibition, Rhizopus stolonifer, Penicillium chrysogenum Introduction Cleome is a dicot genus of the family Cleomaceae. Cleome ciliata Schum. & Thonn. is an annual herb that has digitately compound leaves with leaflets lanceolate to obovate, mostly three, acute at their apices; the stems are not prickly; the fruit is between 2.5 – 6 cm long and the flowers are white, lilac or pink in color (Crocker, 2000). Microorganisms that form a parasitic association with other organisms are classified as pathogens. Some bacteria and fungi can be pathogenic while some are not. Pathogenic bacteria are the major cause of human death and disease while harmful fungi cause infections in healthy persons or immunosuppressed patients. The search for novel potent plants and plant components against pathogenic microorganisms has become increasingly important, as a result of the development of antibiotic resistance in microbes (Ahmad, Aqil, and Owais, 2006). These days, researchers are investigating a wide range of indigenous medicinal plants for antimicrobial active principles with little or no side effects (Chandra et al., 2017). Secondary metabolites are a group of phytochemicals that are by-products of plant metabolism, and are not involved in their vital metabolic pathways. The medicinal quality of plants depends on their chemical constituents that have physiological activity in human systems (Kumar et al., 2009). Antimicrobial investigation of plant extracts and phytochemicals is the basis for antimicrobial drug discovery (Cseke et al., 2006). In a previous study, the phytochemical properties of parts of C. ciliata were determined (Ezeabara and Nwafulugo, 2015), but there is a need to evaluate the biological actions of the phytochemicals present, hence this study. The aim and objective of this study therefore, was to examine the antimicrobial potential of phytochemicals present in parts of C. ciliata. Materials and methods Sources of materials The root, stem and leaf of mature C. ciliata at the flowering stage were collected in May, from the Eastern Farm of the National Root Crop Research Institute, Umudike, Abia State, Nigeria. The plant was identified by Dr C.A. Ezeabara, a Plant Taxonomist in the Department of Botany, Nnamdi Azikiwe University. The voucher specimens were deposited in the herbarium of the same department. Preparation of samples The fresh plant parts were cut into pieces with a knife and then oven-dried at a temperature of 65°C for 12 hours. The samples were ground in a mortar with a pestle, and then in a blender into powdered form. The powdered samples were stored in an air-tight container prior to analyses. Quantitative phytochemical analysis Alkaloids, anthraquinones, flavonoids, phenols, saponins, sterols, terpenoids and tannins were quantitatively determined in the leaf, stem and root extracts of C. ciliata by standard methods (Trease and Evans, 2002). Microbial analysis Preparation of microorganisms for the experiment The pure cultures of the microorganisms were obtained from the Department of Microbiology, Biotechnology and Nuclear Agriculture Research Institute of the Ghana Atomic Energy Commission, Accra, Ghana. The bacteria isolates included Gram-positive Staphylococcus aureus and Streptococcus pneumoniae, and Gram-negative Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi and Shigella sonnei. The fungi used for testing were Aspergillus niger, Fusarium oxysporum, Penicillium chrysogenum and Rhizopus stolonifer. They were collected based on their clinical and pharmacological importance. Isolation and characterization of bacteria and fungi A measured 100 ml of distilled water was poured in a sterile bottle. A soil sample of 1 g was weighed out and added to the bottle of distilled water. The bottle was tightly covered with a lid and thoroughly shaken. One ml of the solution was aseptically and serially diluted. A 1-ml aliquot of a six-fold dilution was plated using potato dextrose agar (PDA) and Nutrient Agar for fungi and bacteria respectively. The plates were gently rotated to distribute the inoculums evenly in the plates and left to solidify under laminar airflow. Each of the inoculated plates was incubated at 37°C for 48 hours. After good growth of the colonies, distinct colonies were sub-cultured with fresh PDA and Nutrient Agar respectively using the streak plate technique and incubated again at 37°C for 72 hours. Each fungal colony was confirmed as the correct species by examining it under a light microscope (DM3000, Germany) using an oil immersion objective after staining with lactophenol in cotton blue dye, while the bacterial colonies were Gram-stained before viewing. Pure isolates were placed on PDA/nutrient agar slants and stored at 4°C until needed. The isolates were characterized based on their cultural and biochemical characteristics. Antimicrobial susceptibility testing The agar well diffusion method was used to determine the sensitivity to the plant extract, as outlined by the International Commission on Microbiological Specifications for Foods (ICMSF, 1998). The pure strains of bacteria were first grown in nutritional broth for 18 hours before use. They were later sub-cultured in Mueller-Hinton agar (Oxoid Ltd). Thirty-eight grams (38 g) of the medium was suspended in 1 litre of distilled water. The medium was completely dissolved by heating it with constant stirring and then boiled for 1 minute. The mixture was autoclaved at 121°C for 15 minutes and cooled at room temperature. Cooled Mueller-Hinton agar was poured into sterile petri dishes on a level laboratory bench and a uniform depth of 4 mm was obtained. The petri dishes were then left to solidify at room temperature. The prepared Mueller-Hinton agar was checked to ensure that the final pH was 7.3 ± 1 at 25°C. The wells were then bored into the agar medium using a sterile 6 mm cork borer. The wells were then filled up with the solution of the extract without allowing it to spill on the surface of the medium. The plates were allowed to stand on the laboratory bench for 1 hour before incubating them at 37°C for 24 hours. The plates were later observed for the zones of inhibition. The degree of inhibition was presented in terms of diameter of the inhibition zone as measured with a transparent ruler. The effect of the extract on bacteria and fungi was compared with that of a standard antibiotic; ampicillin and fungabacter for bacteria and fungi respectively at a concentration of 100 mg/ml. Statistical analysis SPSS version 21 was used to analyse the data. All data were checked for normality and homogeneity using the Kolmogorov–Smirnov and Leven median tests, respectively. Analysis of variance (F-test) was then applied, and Duncan’s multiple range test (DMRT) was used to measure the significance of any differences. The data were presented as mean ± standard error and mean ± standard deviation of three determinations for phytochemical and antimicrobial investigations respectively. Results and discussion The phytochemical composition of the extracts is shown in Table 1. This indicates that C. ciliata contains a wide range of phytochemicals. High concentrations of alkaloids, flavonoids, saponins, terpenoids and tannins were found in the leaf, stem and root. Moreover, the alkaloid, flavonoid, saponin and tannin contents of the C. ciliata parts that were measured in this study were higher than the levels reported by Ezeabara and Nwafulugo (2015). These are the classes of plant secondary metabolites that are most commonly present in plants. The variation in the levels of the phytochemicals could be a result of a number of factors. They could be dependent on location, season, as well as method of extraction. Ezeabara and Egwuoba (2016) reported that the synthesis and accumulation of phytochemicals depend on species, age, season and climatic factors. Phytochemicals accumulate in plant organs such as fruits, flowers, leaves, stems, and roots. Alkaloids (Zulak et al., 2006) and flavonoids (Crozier, Jaganath, and Clifford, 2006) play a defensive role in the plant against herbivores and pathogens. Table 1. Mean quantitative phytochemical composition of the leaf, stem and root of Cleome ciliata Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Results are in mean ± SE of triplicate determinations. The same letter in a column means that the respective means were not significantly different (p > 0.05). Open in new tab Table 1. Mean quantitative phytochemical composition of the leaf, stem and root of Cleome ciliata Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Composition (mg/100 g) . Plant parts . Leaf . Stem . Root . Alkaloids 1.41 ± 0.01b 1.24 ± 0.00a 1.59 ± 0.01c Phenols 0.18 ± 0.00a 0.65 ± 0.00c 0.34 ± 0.02b Steroids 0.50 ± 0.00b 0.29 ± 0.00a 0.27 ± 0.02a Anthraquinones 0.74 ± 0.04c 0.61 ± 0.01b 0.54 ± 0.03a Saponins 1.52 ± 0.00b 0.83 ± 0.00a 1.74 ± 0.02c Terpenoids 1.73 ± 0.01a 1.24 ± 0.01b 1.93 ± 0.05c Tannins 1.17 ± 0.01a 1.09 ± 0.00a 1.27 ± 0.04b Flavonoids 1.83 ± 0.04b 1.53 ± 0.01a 1.74 ± 0.03b Results are in mean ± SE of triplicate determinations. The same letter in a column means that the respective means were not significantly different (p > 0.05). Open in new tab The high levels of these compounds in this study are most likely due to the solvent used in the extraction. In the earlier study (Ezeabara and Nwafulugo, 2015), ethanol was used for alkaloid and saponin extraction while water was used for tannin extraction. The higher concentrations in the current study suggest that methanol is a better solvent for the extraction of these phytochemicals and probably the major factor contributing to the detection of high levels of these biochemicals in C. ciliata. In the antimicrobial tests of the leaf, stem and root extracts of C. ciliata, the plant exhibited inhibitory activity against all test bacterial and fungal pathogens (Tables 2 and 3 and Figs 1–6). This revealed that compounds with antibacterial and antifungal actions are present in C. ciliata. Moreover, the result showed that the leaf extract had the highest inhibitory activity against K. pneumoniae, S. pneumoniae and S. aureus. It also showed high inhibitory activity against P. aeruginosa and S. typhi.Ezeabara and Egenti (2018) reported that a methanol leaf extract of Sida acuta Burm. f. at high concentration showed high inhibitory activity against P. aeruginosa (13.34 ± 1.82 (mm) ± SD). Pseudomonas is a common cause of urinary tract and wound infections where it causes acute or chronic infections (Ekhaise and Anyansi, 2005) while S. typhi is the causative agent for typhoid fever or enteric fever (Pavendan, Anbu, and Sebastian, 2011). The inhibitory activity against these microorganisms may be due to the presence of high levels of alkaloids, anthraquinones, flavonoids, saponins, sterols, terpenoids and tannins (1.41 ± 0.01, 0.74 ± 0.04, 1.83 ± 0.04, 1.52 ± 0.00, 0.50 ± 0.00, 1.73 ± 0.01 and 1.17 ± 0.01 mg/100 g respectively) in the leaf. Doss, Mubarack, and Dhanabalan (2009) reported that tannins isolated from Solanum trilobatum L. exhibited antibacterial activities against Escherichia coli, Pseudomonas aeruginosa, S. typhi and S. aureus. Moreover, some anthraquinones extracted from plants have been reported as therapeutic agents (Kanokmedhakul, Kanokmedhakul, and Phatchana, 2005) while flavonoids have a wide range of biological and pharmacological applications in the human system, probably as a result of their antimicrobial activities. Adedapo, Mogbojuri, Emikpe (2009) reported that saponins are of great pharmaceutical importance because of their relationship to compounds such as the sex hormones, steroids, vitamin D and cardiac glycosides. In addition, Sathya et al. (2012) reported that a terpenoid compound of Tridax procumbens showed antibacterial activity against both Gram-positive and Gram-negative bacteria. These findings imply that the leaf of C. ciliata will serve as a potential source of raw material in the pharmaceutical industry and in modern medicine for the treatment of human diseases caused by these bacterial pathogens. Table 2. Bacterial growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Results are in mean ± std of three replicates. Different letters in a column are significantly different (p < 0.05). Open in new tab Table 2. Bacterial growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Concentration (%) . Bacterial strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Staphylococcus aureus 16.50 ± 0.00d 5.20 ± 0.00c 3.73 ± 0.04a 4.60 ± 0.00b Salmonella typhi 17.25 ± 0.35d 4.23 ± 0.04c 3.00 ± 0.00a 3.73 ± 0.04b Escherichia coli 15.00 ± 0.00d 3.50 ± 0.00c 2.10 ± 0.14a 3.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14d 5.40 ± 0.00c 4.20 ± 0.00a 4.73 ± 0.04b Shigella sonnei 15.61 ± 0.01d 4.50 ± 0.00c 3.30 ± 0.00a 4.33 ± 0.04b Streptococcus pneumoniae 17.50 ± 0.00c 5.40 ± 0.00b 4.73 ± 0.04a 5.35 ± 0.21b Klebsiella pneumoniae 20.43 ± 0.04d 6.20 ± 0.00c 4.80 ± 0.00a 5.80 ± 0.00b 150 Staphylococcus aureus 16.50 ± 0.00d 7.63 ± 0.04c 5.50 ± 0.00a 7.20 ± 0.00b Salmonella typhi 17.25 ± 0.35d 6.80 ± 0.00c 4.51 ± 0.13a 6.08 ± 0.11b Escherichia coli 15.00 ± 0.00d 5.63 ± 0.04c 3.30 ± 0.00a 4.91 ± 0.01b Pseudomonas aeruginosa 16.30 ± 0.14d 7.50 ± 0.00c 6.20 ± 0.00a 7.30 ± 0.00b Shigella sonnei 15.61 ± 0.01d 7.00 ± 0.00c 4.71 ± 0.01a 6.77 ± 0.10b Streptococcus pneumoniae 17.50 ± 0.00c 7.74 ± 0.06b 7.13 ± 0.04a 7.81 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04d 8.00 ± 0.00c 6.00 ± 0.00a 7.20 ± 0.00b 200 Staphylococcus aureus 16.50 ± 0.00d 9.73 ± 0.04c 7.71 ± 0.01a 8.71 ± 0.01b Salmonella typhi 17.25 ± 0.35d 8.48 ± 0.04c 5.50 ± 0.00a 7.80 ± 0.00b Escherichia coli 15.00 ± 0.00d 8.63 ± 0.04c 4.50 ± 0.00a 6.73 ± 0.04b Pseudomonas aeruginosa 16.30 ± 0.14c 11.00 ± 0.00b 9.41 ± 0.01a 9.50 ± 0.00a Shigella sonnei 15.61 ± 0.01d 9.83 ± 0.04c 6.50 ± 0.00a 9.00 ± 0.00b Streptococcus pneumoniae 17.50 ± 0.00d 9.45 ± 0.07b 8.70 ± 0.00a 9.70 ± 0.00c Klebsiella pneumoniae 20.43 ± 0.04d 10.25 ± 0.00a 10.92 ± 0.00b 12.30 ± 0.00c 250 Staphylococcus aureus 16.50 ± 0.00d 12.90 ± 0.00c 10.50 ± 0.00a 11.63 ± 0.04b Salmonella typhi 17.25 ± 0.35d 11.80 ± 0.00c 10.23 ± 0.04a 11.00 ± 0.00b Escherichia coli 15.00 ± 0.00d 11.60 ± 0.00c 9.20 ± 0.00a 10.10 ± 0.14b Pseudomonas aeruginosa 16.30 ± 0.14c 12.20 ± 0.00b 11.87 ± 0.04a 11.71 ± 0.01a Shigella sonnei 15.61 ± 0.01b 10.58 ± 1.03a 10.74 ± 0.02a 10.50 ± 0.00a Streptococcus pneumoniae 17.50 ± 0.00d 13.20 ± 0.00c 12.60 ± 0.00a 13.01 ± 0.01b Klebsiella pneumoniae 20.43 ± 0.04c 13.48 ± 0.04b 12.60 ± 0.28a 13.60 ± 0.00b Results are in mean ± std of three replicates. Different letters in a column are significantly different (p < 0.05). Open in new tab Table 3. Fungal growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Results are in mean ± std of three different determinations. The same letter in a column is not significantly different (p > 0.05). Open in new tab Table 3. Fungal growth inhibition zones of methanol extract of Cleome ciliata Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Concentration (%) . Fungal strains . Mean Zone of Inhibition (mm) ± SD . Control . Leaf . Stem . Root . 100 Aspergillus niger 19.00 ± 0.00d 5.63 ± 0.04c 4.50 ± 0.00a 5.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 5.91 ± 0.01a 6.20 ± 0.00b 6.41 ± 0.01c Rhizopus stolonifer 20.00 ± 0.00c 5.41 ± 0.01a 5.80 ± 0.00b 5.80 ± 0.00b Fusarium oxysporum 21.00 ± 0.00d 4.90 ± 0.00b 4.01 ± 0.01a 6.20 ± 0.00c 150 Aspergillus niger 19.00 ± 0.00d 7.80 ± 0.00c 6.21 ± 0.01a 7.40 ± 0.00b Penicillium chrysogenum 18.50 ± 0.00d 6.50 ± 0.00a 8.76 ± 0.01c 8.50 ± 0.00b Rhizopus stolonifer 20.00 ± 0.00d 7.50 ± 0.00a 7.82 ± 0.02b 9.00 ± 0.00c Fusarium oxysporum 21.00 ± 0.00d 8.73 ± 0.04c 5.23 ± 0.04a 7.80 ± 0.00b 200 Aspergillus niger 19.00 ± 0.00d 9.25 ± 0.07b 8.40 ± 0.00a 9.80 ± 0.00c Penicillium chrysogenum 18.50 ± 0.00d 8.43 ± 0.04a 11.41 ± 0.01b 12.30 ± 0.14c Rhizopus stolonifer 20.00 ± 0.00d 10.23 ± 0.04a 10.50 ± 0.00b 11.91 ± 0.01c Fusarium oxysporum 21.00 ± 0.00d 11.86 ± 0.09c 7.90 ± 0.00a 10.58 ± 0.18b 250 Aspergillus niger 19.00 ± 0.00d 11.74 ± 0.00a 11.80 ± 0.00b 12.61 ± 0.01c Penicillium chrysogenum 18.50 ± 0.00d 12.88 ± 0.03a 13.50 ± 0.00b 14.60 ± 0.00c Rhizopus stolonifer 20.00 ± 0.00d 13.50 ± 0.00c 12.75 ± 0.07a 13.23 ± 0.04b Fusarium oxysporum 21.00 ± 0.00d 13.00 ± 0.00c 11.87 ± 0.04a 12.80 ± 0.00b Results are in mean ± std of three different determinations. The same letter in a column is not significantly different (p > 0.05). Open in new tab Figure 1. Open in new tabDownload slide Zone of Inhibition (mm) of bacterial pathogens by leaf extract of Cleome ciliata. Figure 2. Open in new tabDownload slide Zone of Inhibition (mm) of bacterial pathogens by stem extract of Cleome ciliata. Figure 3. Open in new tabDownload slide Zone of Inhibition (mm) of bacterial pathogens by root extract of Cleome ciliata. Figure 4. Open in new tabDownload slide Zone of Inhibition (mm) of fungal pathogens by leaf extract of Cleome ciliata. Figure 5. Open in new tabDownload slide Zone of Inhibition (mm) of fungal pathogens by stem extract of Cleome ciliata. Figure 6. Open in new tabDownload slide Zones of Inhibition (mm) of fungal pathogens by root extract of Cleome ciliata. The root extract of C. ciliata showed the highest inhibitory activity against K. pneumoniae and S. pneumoniae. This may be due to the high levels of alkaloids, saponins, terpenoids and tannins in the root. Organisms like S. pneumoniae have been associated with many human infections while K. pneumoniae has been found to account for up to 55% of nosocomial infections in some parts of Nigeria (Okonko et al., 2008). Moreover, the sensitivity and susceptibility of the test pathogens to the plant extracts was found to vary among the plant extracts. In particular, K. pneumoniae and S. pneumoniae were highly sensitive and susceptible to the plant extracts while Escherichia coli was the least susceptible. This suggested that E. coli is resistant to the inhibitory effects of the plant extract. Escherichia coli is associated with cholera, dysentery, gastroenteritis, typhoid fever and urinary tract infections (Orji, Ezenwaje, and Anyaegbunam, 2006). The methanol extracts of the leaf, stem and root of C. ciliata inhibited all the test fungi. Although the response of the test fungi varied among the plant extracts, the antagonistic effects of the leaf and root extracts were more evident than those of stem. This could be attributed to the higher concentration of bioactive compounds in the leaf and root than the stem. This implies that the leaf and root extracts of C. ciliata will serve as a better source of raw materials for the synthesis of new drugs that could be used to control and treat human and animal diseases, since A. niger, F. oxysporum, P. chrysogenum and R. stolonifer have all been associated with various diseases in humans and animals. Both the antibacterial and antifungal properties of the leaf, stem and root extracts of C. ciliata were found to increase with increasing concentration of the extract. This indicates that the antimicrobial actions of C. ciliata parts are dose dependent and that the three different parts have antimicrobial activities that can be fully utilized in phytomedicine. However, the standard antibiotics, ampicillin and fungabacter for bacteria and fungi, respectively, that were used as the standard control, showed higher inhibitory activities against all test bacteria and fungi. The superiority of conventional antibiotics over plant extracts has been extensively reported (Kubmarawa, Ajoku, and Okrie, 2002; Doss, Mubarack, and Dhanabalan, 2009; Samaila et al., 2014). This suggests that plant extracts need to be enhanced for better performance in health care delivery. Conclusion This study revealed that the methanol leaf extract contained the highest concentrations of anthraquinones and flavonoids while the root contained the highest concentrations of alkaloids, saponins, tannins and terpenoids. Moreover, they exhibited antibacterial and antifungal properties that were dose-dependent. The leaf extract showed the highest inhibition of K. pneumoniae, S. pneumoniae, S. aureus, S. typhi, P. aeruginosa, R. stolonifer and F. oxysporum while the root extract showed the highest inhibitory activity against K. pneumoniae and P. chrysogenum. Hence, these parts of C. ciliata will be very effective for the treatment of the diseases caused by these organisms. C. ciliata thus contains phytochemicals with antimicrobial properties, suggesting its potential use in the formulation of novel drugs. Author’s biography I.C.O. graduated with a Second Class Honours Upper Division in BSc Botany from Nnamdi Azikiwe University, Awka, Nigeria. His interest is in Phytomedicine. He performed the experiments. Dr C.A.E. designed and supervised the study and assisted in writing of the paper. References Adedapo , A. A. , Mogbojuri , O. M. and Emikpe , B. O. ( 2009 ) Safety evaluation of the aqueous extract of the leaves of Moringa oleifera , Journal of Medical Plants Research , 3 ( 8 ), 586 – 592 . Google Scholar OpenURL Placeholder Text WorldCat Ahmad , I. , Aqil , F. and Owais , M. ( 2006 ) Modern Phytomedicine: Turning Medicinal Plants into Drugs , Wiley-VCH , India , p. 404 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Chandra , H. , Bishnoi , P., Yadav , A. et al. . ( 2017 ) Antimicrobial resistance and the alternative resources with special emphasis on plant-based antimicrobials—a review , Plants , 6 ( 16 ), 11 . Google Scholar OpenURL Placeholder Text WorldCat Crocker , W. ( 2000 ) Growth of Plants , Reinhold Publishing Corporation, Ltd , London , p. 459 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Crozier , A. , Jaganath , I. B. and Clifford , M. N. ( 2006 ) Phenols, polyphenols and tannins: an overview, in Crozier A., Clifford M. N. and Ashihara H. (eds), Plant Secondary Metabolites: Occurrence, Structure and Role in the Human Diet , Blackwell Publishing Limited , Oxford, UK , pp. 1 – 24 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Cseke , L. J. , Kirakosyan , A., Kaufman , P. B. et al. . ( 2006 ) Natural Product from Plants , 2nd edn, CRC Press, Taylor & Francis Group, L.L.C , Boca Raton, Florida , p. 611 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Doss , A. , Mubarack , H. M. and Dhanabalan , R. ( 2009 ) Antibacterial activity of tannins from the leaves of Solanum trilobatum Linn , Indian Journal of Science and Technology , 2 ( 2 ), 41 – 43 . Google Scholar OpenURL Placeholder Text WorldCat Ekhaise , F. O. and Anyansi , C. C. ( 2005 ) Influence of brewery effluent discharge on the microbiological and physicochemical quality of Ikpoba River, Nigeria , African Journal of Biotechnology , 4 ( 10 ), 1062 – 1065 . Google Scholar OpenURL Placeholder Text WorldCat Ezeabara , C. A. and Egenti , M. O. ( 2018 ) Phytochemical and antimicrobial investigations on various parts of Sida acuta Burm. f , Journal of Ayurvedic and Herbal Medicine , 4 ( 2 ), 71 – 75 . Google Scholar OpenURL Placeholder Text WorldCat Ezeabara , C. A. and Egwuoba , G. C. ( 2016 ) Comparative screening of phytochemical and proximate constituents of leaf, stem and root of Oldenlandia corymbosa L. and Oldenlandia herbacea (L.) Roxb , American Journal of Life Science Research , 4 ( 3 ), 113 – 118 . Google Scholar OpenURL Placeholder Text WorldCat Ezeabara , C. A. and Nwafulugo , S. N. ( 2015 ) Comparison of phytochemical and proximate compositions of parts of Cleome ciliata Schum. & Thonn. and Cleome viscosa L. , World Journal of Biomedicine and Pharmaceutical Sciences , 1 ( 1 ), 1 – 5 . Google Scholar OpenURL Placeholder Text WorldCat International Commission on Microbiological Specifications for Foods . ( 1998 ) Principles for the establishment of microbiological food safety objectives and related control measures , Food Control , 9 ( 6 ), 379 – 384 . Crossref Search ADS WorldCat Kanokmedhakul , K. , Kanokmedhakul , S. and Phatchana , R. ( 2005 ) Biological activity of anthraquinones and triterpenoids from Prismatomeris fragrans , Journal of Ethnopharmacology , 100 ( 3 ), 284 – 288 . Google Scholar Crossref Search ADS PubMed WorldCat Kubmarawa , D. , Ajoku , G. and Okrie , D. A. ( 2002 ) Antimicrobial spectrum of hexane of Commiphora kerstingii Engl. (Buseraceae) , Technology Development , 8 , 29 – 32 . Google Scholar OpenURL Placeholder Text WorldCat Kumar , A. , Rajput , G., Dhatwalia , V. K. et al. . ( 2009 ) Phytocontent screening of Mucuna seeds and exploit in opposition to pathogenic microbes , Journal of Biological and Environmental Sciences , 3 , 71 – 76 . Google Scholar OpenURL Placeholder Text WorldCat Okonko , I. O. , Adejoye , O. D., Ogunnusi , T. A. et al. . ( 2008 ) Urban Domestic Solid Waste Management, Nimo , Rex Charles and Patrick Limited , Awka , pp. 5 – 7 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Orji , M. U. , Ezenwaje , E. E. and Anyaegbunam , B. C. ( 2006 ) Spatial appraisal of shallow well water pollution in Awka, Nigeria , Nigeria Journal of Microbiology , 20 ( 3 ), 1384 – 1389 . Google Scholar OpenURL Placeholder Text WorldCat Pavendan , P. , Anbu , S. S. and Sebastian , R. C. ( 2011 ) Physico Chemical and microbial assessment of drinking water from different water sources of Tiruchirappalli District, South India , European Journal of Experimental Biology , 1 ( 1 ), 183 – 189 . Google Scholar OpenURL Placeholder Text WorldCat Samaila , A. B. , Yarma , A. A., Oshomoh , E. O. et al. . ( 2014 ) Antimicrobial activity of the leaf and root extracts of Kigelia africana and Albizia chevalieri against Staphylococcus aureus , Greener Journal of Microbiology and Antimicrobials , 3 ( 1 ), 001 – 007 . Google Scholar Crossref Search ADS WorldCat Sathya , B. S. , Jayasurya , K. S., Sankaranarayanan , S., Bama , P. ( 2012 ) Antibacterial activity of different phytochemical extracts from the leaves of Tridax procumbens Linn.: identification and mode of action of the terpenoid compound as antibacterial , International Journal of Pharmacy and Pharmaceutical Sciences , 4 ( Suppl 1 ), 557 – 564 . Google Scholar OpenURL Placeholder Text WorldCat Trease , G. E. and Evans , W. C. ( 2002 ) Pharmacognosy , 15th edn, W.B. Saunders , London , p. 406 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Zulak , K. G. , Liscombe , D. K., Ashihara , H. et al. . ( 2006 ) Alkaloids, in Crozier A., Clifford M. N. and Ashihara H. (eds), Plant Secondary Metabolites: Occurrence, Structure and Role in the Human Diet , Blackwell Publishing Limited , Oxford, UK , pp. 102 – 136 . Google Scholar Google Preview OpenURL Placeholder Text WorldCat COPAC Author notes Supervisor: Dr Chinelo A. Ezeabara, Department of Botany, Nnamdi Azikiwe University, PMB 5025 Awka, Nigeria. © The Author(s) 2019. Published by Oxford University Press. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com © The Author(s) 2019. Published by Oxford University Press.

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BioScience HorizonsOxford University Press

Published: Jan 1, 2018

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