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Antibacterial Potential of Capparis spinosa and Capparis decidua Extracts.

Byline: Tehseen Gull, Bushra Sultana, Ijaz Ahmad Bhatti and Amer Jamil

Abstract

Capparis spinosa and C. decidua are indigenous medicinal plants of Pakistan which are rich in antioxidant compounds. In present investigation, the aqueous menthanolic, ethanolic and acetone extracts of stem bark, shoot, fruit, flower and roots of both species were investigated for their antimicrobial potential in comparison with amoxicillin and ciprofloxacin (control). The effect of these extracts was evaluated on the growth of four bacteria i.e., Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Pasteurella multocida using disc diffusion and minimum inhibitory concentration (MIC) assays. After comparing all extracts, it was found that methanolic extracts of selected parts of both species significantly reduced the growth of all four bacteria although different extracts expressed varying efficacy in reducing bacterial growth.

It was further noted that S. aureus growth was mainly inhibited by methanolic extracts of C. decidua flowers and roots (GIZ: 26.5 and 26.4 mm, respectively), E. coli population was effectively reduced by methanolic extracts of C. decidua roots and shoots (GIZ: 29.1 26.7 mm, respectively) while B. subtilis growth was inhibited maximally by methanolic extracts of C. spinosa stem bark and C. decidua fruits (GIZ: 26.8 and 26.8, respectively). Moreover, methanolic extract of C. decidua root markedly inhibited the growth of P. multocida (25.7 mm). On the basis of above mentioned findings, it can safely be concluded that C. decidua possess higher antimicrobial potential than C. spinosa and these can be used as natural antibacterial agent.

Keywords: Acetone; Bacillus subtilis; Escherichia coli; Ethanol; Methanol; Pasteurella multocida; Staphylococcus aureus

Introduction

The medicinal plants have been used traditionally in different civilization since ancient times due to their strong therapeutic effects on human beings. Recently, medicinal plants are acquiring good place in pharmaceutical industries owning higher nutritional quality and medicinal compounds which are capable for curing diseases (Shrishailappa et al., 2001). These are also used against microbial infections for treating ailments due to the presence of antimicrobial compounds (Iwu et al., 1999). In this modern era, the pharmacists and physicians mostly prescribe drugs discovered, isolated and manufactured synthetically which allow a fast recovery from diseases but, as their use increases, the side effects for the host and the resistance of infecting agents to these products also increase. Synthetic drugs have been preferred for over decades but now the use of the herbal medicines is being preferred to avoid blind dependency on synthetic ones.

Recently, many plants have been explored and reported for their ability to inhibit the growth of microorganisms, which can replace the chemically manufactured drugs (Ilic et al., 2004; Mandalari et al., 2007). Consequently, plant based compounds like isoflavones, gamma thionin and homoisoflavinoids have been discovered and studied extensively (Hong et al., 2006; Franco et al., 2006; Mhaeswara et al., 2006). Additionally, sulfur rich plant extracts have been studied and their potential against bacterial pathogens has been reported (Iscan et al., 2002; Kalemba and Kunicka, 2003).

Many plant species have been reported for their medicinal uses around the world. According to a safe estimate, 6000 plant species have been reported in Pakistan (Shinwari et al., 2000). Many of these are traditionally used as therapy for curing and preventing different diseases. These plants belong to different families and genera including higher plants, trees, herbs, shrubs and grasses. Capparis species belonging to family Capparaceae are not exceptions for their medicinal attributes. This family consists of 250 species, out of which C. spinosa and C. decidua are native to Pakistan which have been reported for their traditional uses in folk medicine (Mabberley, 1997; Hamed et al., 2007). The aerial parts of these species have been reported as a potential source to restrain the bacterial growth.

The extracts of fermented C. spinosa parts were reported to be effective to inhibit growth of different bacterial strains especially those which have acquired resistance to drugs like ciprofloxacin, vancomycin and teicoplanin (Perez et al., 2005; 2006a, b). Based on these findings, it is a dire need to explore the antimicrobial potential of different parts of C. spinosa and C. decidua as previously, no investigation has been carried out in Pakistan on these plants. So, the present study was designed aiming at exploring the antibacterial potential of aqueous methanol, aqueous ethanol and aqueous acetone extracts against Gram positive and negative bacteria.

Materials and Methods

Sample Collection

Five different plants of each species (C. spinosa and C. decidua) were selected and tagged from Cholistan desert, Bahawalpur region of Punjab. The samples of stem bark, shoots, fruits, flowers and roots of the both species were collected from the selected plants. The samples were further identified and authenticated by Dr. M. Hameed, Taxonomist, University of Agriculture Faisalabad, Pakistan. Average temperature (27C), dew point (16C) and relative humidity (47%) were recorded in sampling season (April).

Preparation of Extracts

The plant samples were ground to pass 2 mm sieve and the powder plant samples were separately extracted with three different solvents aqueous methanol (methanol: water, 80:20 v/v), aqueous ethanol (ethanol: water, 80:20 v/v) and aqueous acetone (acetone: water, 80:20 v/v). Dry powdered samples of each plant part (10 g) were mixed separately with 100 mL of each solvent for 6 h at room temperature in an orbital shaker (Pamico, Pak). The extract and residues were separated using Whatman's filter paper No. 1. The residues were extracted twice with the respective fresh solvents, and the obtained three extracts were pooled. The combined extracts were evaporated under reduced pressure at 45 C, using rotary evaporator (EYELA, N-N Series). The crude concentrated extracts were stored in refrigerator at - 4C, until used for further studies.

Bacterial Strains

The pure cultures of Bacterial strains (Staphylococcus aureus: NCTC 6571, Escherichia coli: ATCC 8739, Bacillus subtilis: NCTC 10400 and P. multocida: wild type) were obtained from Department of Biochemistry, University of Agriculture Faisalabad, Pakistan and Nuclear Institute for Agriculture and Biology (NIAB), Faisalabad, Pakistan which were further characterized from Institute of Microbiology, University of Agriculture Faisalabad, Pakistan. The bacteria were cultured overnight at 37C on nutrient agar (NA). Antibacterial activity of 80% aqueous methanolic, ethonolic and acetone extracts of selected parts of C. spinosa and C. decidua were individually evaluated against the above bacteria by following two methods.

a. Disc diffusion method (NCCLS, 1997). Bacterial suspension (100 L) containing 108 colony-forming units (CFU)/mL was spread on petri plates (diameter A- height: 150 A- 25 mm) containing NA medium (50 mL media/plate). The wick paper discs (6 mm in diameter, 11 discs in each petri plate including two positive and one negative control) were separately saturated with 15 L of each extract placed on the agar which had previously been inoculated with the selected test bacteria. Amoxycillin and ciprofloxacin were used as positive references for bacteria. Discs without samples were used as a negative control. Plates were kept at 4C for 1 h and incubated at 37C for 24 h. Antibacterial activity was assessed by measuring the diameter of the growth-inhibition zone (GIZ) in mm (including disc diameter of 6 mm) for the test organisms comparing to the controls.

. Resazurin microtitre-plate assay. Modified resazurin microtitre-plate assay was also used for the measurement of MIC (minimum inhibitory concentration) of test bacteria against different extracts (Sarker et al., 2007). Briefly, 100 L extract and standard drug (1 mg/mL in methanol) was pipetted into the first row of the 96 well plates. To all other wells 50 L of nutrient broth was added. Two fold serial dilutions were performed using a multichannel pipette such that each well had 50 L of the test material in serially descending concentrations. Then, 30 L of 3.3A- strength isosensitised broth and 10 L of resazurin indicator solution (prepared by dissolving 270 mg tablet in 40 mL of sterile distilled water) were added in each well. Finally, 10 L of bacterial suspension was added to each well to achieve a concentration of approx 5 A- 105 CFU mL-1.

Each plate had a set of controls: a column with ciprofloxacin and second with amoxicillin as positive control, a column with all solutions with the exception of the test compound, a column with all solutions with the exception of the bacterial solution adding 10 L of nutrient broth instead and a column with methanol as a negative control. The plates were prepared in triplicate. Plates were enfolded loosely with cling film and incubated at 37 C for 24 h. The color change was then assessed visually. The growth was indicated by color changes from purple to pink or colorless. The lowest concentration at which colour change occurred was taken as the MIC value.

Statistical Analysis

Each parameter was analyzed individually in triplicate and data was reported as mean standard error. Data were analyzed by analysis of variance (ANOVA) using Minitab 2000 Version 13.2 statistical software (Minitab Inc. Pennysalvania, USA) at 5% significance level.

Results

The growth of all four bacteria was significantly affected by the application of plant extracts. It was found that aqueous methanolic extract of the stem bark of C. spinosa effectively inhibited the growth of all

Table 1: Antimicrobial activity (mm) of stem bark extracts of C. spinosa and C. decidua (assessed by disc diffusion assay)

Plant Species###Solvents###Bacteria

###Staphylococcus aureus Escherichia coli###Bacillus subtilis###Pasteurella multocida

C. spinosa###Methanol###15.70.5b###17.90.7b###26.80.7a###24.70.4a

###Ethanol###11.10.3b###13.20.2b###19.20.2a###12.70.3b

###Acetone###13.20.1bc###18.60.7a###13.70.4bc###15.30.5b

C. decidua###Methanol###20.40.8ab###21.10.6ab###23.40.9a###19.70.7ab

###Ethanol###14.20.6b###14.30.5b###17.10.6b###21.20.9a

###Acetone###13.70.5c###16.90.4b###12.90.5c###19.00.4a

Control###Amoxicillin###31.41.2###32.31.4###31.60.9###30.30.8

###Ciprofloxacin###29.61.1###30.91.2###27.81.3###27.91.3

Table 2: Minimum inhibitory concentration (g mL-1) of stem bark extracts of C. spinosa and C. decidua

Plant Species###Solvents###Bacteria

###Staphylococcus aureus Escherichia coli###Bacillus subtilis###Pasteurella multocida

C. spinosa###Methanol###145.25.1b###184.64.8a###123.25.2c###126.78.4c

###Ethanol###189.14.3b###236.23.7a###159.73.3c###158.36.1c

###Acetone###256.73.6b###278.93.9a###207.86.5c###246.72.2b

###a###b###c

C. decidua###Methanol###236.74.5###212.34.5###128.93.1###199.06.2bc

###Ethanol###241.27.9b###268.95.3a###179.13.9d###213.27.7c

###Acetone###303.46.3a###310.18.1a###233.48.6b###296.77.5a

Control###Amoxicillin###22.21.1ab###13.70.6b###10.80.5b###29.51.4a

###Ciprofloxacin###14.80.7a###10.70.4ab###9.20.4ab###13.40.7a

Table 3: Antimicrobial activity (mm) of shoot extracts of C. spinosa and C. decidua (assessed by disc diffusion assay)

Plant Species###Solvents###Bacteria

###Staphylococcus aureus Escherichia coli###Bacillus subtilis###Pasteurella multocida

C. spinosa###Methanol###21.00.2b###21.90.2b###24.60.5a###23.60.7a

###Ethanol###17.90.2b###13.70.4c###19.90.7a###16.90.3b

###Acetone###11.10.5b###14.50.2a###14.90.3a###12.50.7b

C. decidua###Methanol###19.80.3b###26.70.7a###12.40.4c###20.70.1b

###b###b###a

###Ethanol###16.70.5###15.90.3###19.10.4###11.80.3b

###Acetone###12.90.5c###17.60.6a###14.70.3b###12.60.2c

Control###Amoxicillin###31.51.2###30.61.0###30.61.2###29.30.8

###Ciprofloxacin###30.60.9###29.81.4###28.71.1###27.20.5

selected bacteria but its maximum inhibiting potential was recorded against B. subtilis with greater inhibition zones (26.8 mm) and lower MIC values (123.2 g mL-1) followed by P. multocida (GIZ, 24.7 mm; MIC, 158.3 g mL-1), while aqueous ethanol and acetone extracts of C. spinosa stem bark in that order were effective in inhibiting the corresponding population of B. subtilis and E. coli (GIZ, 19.2, 18.6 mm; MIC, 278.9, 159.7 g mL-1, respectively) as given in Table 1 and 2. In case of C. decidua stem bark, methanolic extract maximally inhibited B. subtilis (GIZ, 23.4 mm; MIC, 128.9 g mL- 1), while its ethanol and acetone extracts showed considerable antibacterial activities with GIZ of 21.2, 19.0 mm and MIC values of 213.2, 296.7 g mL-1 against P. multocida (Table 1 and 2).

Antibacterial activity of aqueous methanolic extract of C. spinosa shoot maximally controlled the population of B. subtilis (GIZ, 24.6 mm; MIC, 148.9 g mL-1) followed by P. multocida while its ethanol and acetone extracts effectively inhibited the growth of B. subtilis and E. coli with appreciable values of inhibition zones (GIZ) 13.7, 19.9 and 14.5, 14.9 mm minimum inhibitory concentrations (MIC) of 136.7, 198.9 and 251.9, 275.6 g mL-1, respectively (Table 3 and 4). The methanolic extract of C. decidua shoot maximally reduced the growth of E. coli followed by P. multocida, while showed less activity against E. coli and S. aureus. The antibacterial activity of aqueous ethanol and acetone extracts of shoot was good against the gram positive and gram negative bacteria in similar trend like C. spinosa shoot extracts (Table 3 and 4).

Aqueous methanol, ethanol and acetone extract of C. spinosa and C. decidua fruits exhibited varying antibacterial activity as shown by the inhibition zones (GIZ) and MIC values (Table 5 and 6). The best activity of methanolic

Table 5: Antimicrobial activity (mm) of fruit extracts of C. spinosa and C. decidua (assessed by disc diffusion assay)

Plant Species###Solvents###Bacteria

###Staphylococcus aureus Escherichia coli###Bacillus subtilis###Pasteurella multocida

C. spinosa###Methanol###17.70.5###20.90.1###23.90.4###24.90.6

###Ethanol###11.90.6###15.20.4###16.10.3###17.70.5

###Acetone###12.60.2###16.70.3###12.70.7###12.10.1

C. decidua###Methanol###21.40.3###24.10.7###26.80.2###21.70.1

###Ethanol###16.90.2###15.30.1###19.20.3###23.70.6

###Acetone###13.20.4###18.90.7###18.70.5###19.90.8

Control###Amoxicillin###30.10.9###32.21.6###34.10.9###29.91.4

###Ciprofloxacin###29.70.7###27.31.3###31.71.2###26.61.0

Table 6: Minimum inhibitory concentration (g mL-1) of fruit extracts of C. spinosa and C. decidua

Plant Species###Solvents###Bacteria

###Staphylococcus aureus Escherichia coli###Bacillus subtilis###Pasteurella multocida

C. spinosa###Methanol###110.92.9b###253.28.4a###126.75.1b###120.12.5b

###Ethanol###125.63.5c###266.74.6a###160.12.2b###176.74.4b

###Acetone###272.74.7b###306.95.1a###253.96.2bc###234.98.2c

C. decidua###Methanol###231.27.9a###158.34.8c###134.25.7cd###191.94.3b

###Ethanol###269.98.3a###188.93.0c###156.72.9d###233.77.9b

###Acetone###316.49.8a###260.17.5c###212.86.8d###285.75.2b

Control###Amoxicillin###21.20.6b###9.70.5c###12.40.6c###29.20.9a

###Ciprofloxacin###14.30.4a###8.90.3ab###6.80.2b###9.70.3ab

Table 7: Antimicrobial activity (mm) of flower extracts of C. spinosa and C. decidua (assessed by disc diffusion assay)

Plant Species###Solvents###Bacteria

###Staphylococcus aureus Escherichia coli###Bacillus subtilis###Pasteurella multocida

C. spinosa###Methanol###23.70.5ab###26.50.7a###17.60.4bc###20.70.8b

###Ethanol###14.80.2###18.20.9###19.70.1###15.90.3

###Acetone###12.10.3b###21.60.7a###11.20.5b###14.70.2b

C. decidua###Methanol###26.50.4a###23.90.1b###19.60.3c###22.30.6b

###Ethanol###19.10.2a###13.40.9b###11.30.5b###13.40.3b

###Acetone###21.30.3a###16.40.5b###13.20.8c###11.30.2c

Control###Amoxicillin###30.91.2###30.20.9###31.61.1###29.70.9

###Ciprofloxacin###28.51.1###29.31.1###30.30.9###27.90.7

Table 8: Minimum inhibitory concentration (g mL-1) of flower extracts of C. spinosa and C. decidua

Plant Species###Solvents###Bacteria

###Staphylococcus aureus Escherichia coli###Bacillus subtilis###Pasteurella multocida

C. spinosa###Methanol###122.11.5c###181.23.1a###111.21.6cd###146.73.8b

###Ethanol###147.82.7bc###215.62.6a###175.63.7b###158.93.6bc

###Acetone###266.76.4a###224.05.3b###196.77.2c###206.77.1c

C. decidua###Methanol###112.33.7 ab###133.43.6a###96.34.2b###112.33.9ab

###Ethanol###143.12.7b###163.42.6a###138.24.2b###133.41.9b

###Acetone###204.53.7d###258.92.6b###295.68.9a###222.32.7c

Control###Amoxicillin###23.10.6b###11.20.4c###9.60.3c###27.20.5a

###Ciprofloxacin###17.40.3a###7.80.2b###7.40.2b###10.80.4b

extract of fruit was observed against P. multocida with large GIZ (24.9 mm) and small MIC values (120.1 g mL-1) values, while its acetone extract maximally reduced the growth of E. coli (GIZ, 16.7 mm; MIC, 3.6 g mL-1). Although the aqueous methanolic extracts of C. decidua fruits exhibited different trend against the selected bacteria, however it maximally reduced the population of B. subtilis and E. coli (GIZ, 26.8, 24.1 mm; MIC, 134.2, 158.3 g mL-1, respectively) while its ethanolic and acetone extracts effectively controlled the population of P. multocida with GIZ of 23.7, 19.9 mm and minimum inhibitory concentration 233.7, 285.7 g mL-1 (Table 5 and 6).

Table 9: Antimicrobial activity (mm) of root extracts of C. spinosa and C. decidua (assessed by disc diffusion assay)

Plant Species###Solvents###Bacteria

###Staphylococcus aureus###Escherichia coli###Bacillus subtilis###Pasteurella multocida

C. spinosa###Methanol###21.10.9###23.90.7###21.70.4###20.40.5

###Ethanol###14.90.4b###19.20.1a###16.10.6ab###13.70.8b

###Acetone###15.60.3###16.70.4###17.70.8###15.10.2

C. decidua###Methanol###26.40.2ab###29.10.4a###23.80.9b###25.70.1ab

###Ethanol###19.90.5ab###22.30.3a###18.20.6ab###23.40.5a

###Acetone###20.20.1a###21.90.6a###15.70.3b###22.90.2a

Control###Amoxicillin###32.11.6###34.21.7###32.11.3###29.71.2

###Ciprofloxacin###31.71.4###32.31.4###29.71.0###27.10.8

Table 10: Minimum inhibitory concentration (g mL-1) of root extracts of C. spinosa and C. decidua

Plant Species###Solvents###Bacteria

###Staphylococcus aureus Escherichia coli###Bacillus subtilis###Pasteurella multocida

C. spinosa###Methanol###156.71.4b###142.66.3b###157.84.2b###207.98.1a

###Ethanol###198.75.6b###194.12.1b###231.44.8a###231.46.6a

###Acetone###277.18.0a###229.67.6b###282.65.3a###264.87.9a

C. decidua###Methanol###182.35.3ab###163.75.7b###197.73.6a###204.86.9a

###b###b###a

###Ethanol###189.34.6###179.64.1###232.34.8###232.77.5a

###Acetone###263.45.5c###221.76.2d###318.17.9a###286.48.1b

Control###Amoxicillin###22.51.1b###7.90.3c###10.80.5c###33.61.6a

###Ciprofloxacin###15.70.7a###8.00.4b###5.60.2c###11.80.5ab

Screening of the antimicrobial activity of the aqueous methanol, ethanol and acetone extract of flowers of C. spinosa and C. decidua was also done against four pathogenic bacteria. The results of disc diffusion assay and measurement of inhibitory concentrations showed variation in the antibacterial activity (Tables 7 and 8). The methanol extract of C. spinosa flower showed stronger antibacterial activity against E. coli followed by S. aureus, P. multocida and B. subtilis. The values of GIZ offered for bacterial stains were 26.5, 23.7, 20.7 and 17.6 mm, respectively. Generally, the tested Gram positive bacteria were found to be more sensitive to flowers extract than Gram negative bacteria. Little differences were observed for the GIZ among these extracts, since all of them showed a strong activity for the strains assayed.

However, more precise results on the antimicrobial properties were obtained through the determination of minimum inhibitory concentration that were found to be 181.2, 122.1, 146.7 and 111.2 g mL-1 against E. coli, P. multocida, S. aureus and B. subtilis, respectively. Ethanolic extract of C. spinosa flower was effective against B. subtilis (GIZ, 19.7 mm; MIC, 175.6 g mL-1) followed by E. coli (GIZ, 18.2 mm, MIC, 215.6 g mL-1) while all three extracts of C. decidua flowers i.e., aqueous methanol, ethanol and acetone effectively reduced the population of S. aureus and E. coli (Table 7 and 8).

The antibacterial effectiveness of aqueous methanol, ethanol and acetone extract of C. spinosa and C. decidua roots was evaluated by the measurement of inhibition zones (GIZ) and minimum Inhibitory concentration (MIC) is in Table 9 and 10. The results from the disc diffusion assay indicated that aqueous methanol extract of C. spinosa root showed comparable antibacterial activity in selected strains of bacteria (E. coli, S. aureus, Bacillus subtilis and P. multocida) with GIZ 23.9, 21.7, 21.2 and 20.4 mm, respectively. The ethanol and acetone extracts of C. spinosa root exhibited best antibacterial activity against E. coli and Bacillus subtilis (GIZ, 19.2, 17.7mm; MIC, 142.6, 282.6 g mL-1, respectively) while methanolic extract of C. decidua roots showed best activity against the E. coli (GIZ, 29.1 mm; MIC, 142.6 g mL-1) and ethanolic and acetone extracts of C. decidua roots reduced the growth of P. multocida (GIZ, 23.4,22.9 mm; MIC, 232.7, 286.4 g mL-1, respectively) (Table 9 and 10).

Discussion

Resistance in bacteria against drugs is intensifying due to their extensive use in clinical medicines. No doubt, the scientists are continuously working on discovering and formulating new drugs to cure microbial infections but drug resistant bacterial strains are continuously opposing a challenge to pharmacists. It is urging for researchers to explore the natural products qualifying antibacterial potential in their extracts. Medicinal plants have been used in since ages but their antimicrobial potential as a source of drugs have not been explored extensively although these are used in folk medicine or indirectly in pharmaceutical drugs. The resistance in pathogenic bacteria against drugs and the traditional practices of using medicinal plants against infectious diseases is accelerating the research work related to explore the efficacy of plant extracts or plant derived phytochemical compounds (Chan et al., 2007; Pitchamuthu et al., 2012).

In present investigation, antibacterial potential of selected parts of C. spinosa and C. decidua was evaluated against four Gram positive and negative bacteria. The growth of all four bacteria was significantly inhibited by the application of Capparis extracts in comparison with synthetic drugs i.e., amoxycillin and ciprofloxacin. Aqueous methanolic extracts were found to be more effective in comparison with ethanol and acetone extracts against all bacteria but extracts of different parts exhibited variable efficacy in limiting bacterial development. Previously, the inhibiting potential of aerial parts of C. spinosa plants has been reported against Gram-positive and negative bacteria (Mahasneh, 2002). In this study, Capparis stem bark aqueous methanolic extract effectively inhibited Bacillus subtilis and P. multocida growth, while other bacteria showed some resistance. The difference in bacterial response against solvents might be due to cell membrane permeability.

Gram negative bacteria have an outer phospholipidic membrane, which makes the wall impermeable to antimicrobial chemical compound, while Gram positive bacteria have peptidoglycan layer, which is permeable to these substances (Sharma et al., 2010). It was further found that aqueous methanolic extract of C. decidua shoot has inhibiting potential against Bacillus subtilis and Aspergillus niger (Iqbal et al., 2006). Upadhyay et al. (2010) reported the antimicrobial potential of absolute and aqueous methanol extracts of C. decidua against Klebsiella pneumoniae, E. coli, Micrococcus luteus, Streptococcus pneumoniae, S. aureus, Bacillus cereus and Lactobacilus acidophilus. Those researchers reported 36 mm growth inhibition zone (GIZ) of S. aureus when induced with C. decidua shoot extract while in the present study 21.0 and 19.8 mm GIZ of S. aureus was recorded by aqueous methanolic extract of C. decidua and C. spinosa shoot extract, respectively.

As, the fruits and flowers of C. spinosa and C. decidua are also rich in antioxidant compounds (Zia-ul-Haq et al., 2011), their extracts exhibited good antimicrobial potential in comparison with synthetic drugs i.e., amoxicillin and ciprofloxacin as found in the present investigation. Previously, C. spinosa flower extracts have been reported as 100 and 90% effective against Gram positive and negative bacteria, respectively (Ibrahim, 2012). Likewise, extracts obtained from fermented caper fruits have been found to contain effective antimicrobial agents against bacterial strains that had become resistant to drugs like ciprofloxacin, vancomycin and teicoplanin (Perez et al., 2005; 2006a, b).

The roots of C. spinosa and C. decidua are not exception to exhibit antibacterial activity. The root extracts of both species exhibited growth inhibiting effect on all studied bacteria (Table 9, 10). The potential of C. spinosa roots has also been studied on Deinococcus radiophilus (Boga et al., 2011). Aqueous methanolic extract of C. decidua roots showed larger GIZ for S. aureus and E. coli, while Mali et al. (2004) found that ethanolic extract of C. decidua roots is more effective against S. aureus and E. coli in comparison with other extracts. This difference might be attributed to extraction technique/solvent, germplasm, climatic factors and growing conditions. A few studies are available for antimicrobial potential of C. spinosa and C. decidua root extracts while the researchers have worked on a few other species' root extracts.

The effectiveness of C. grandiflora root extract was studied against S. aureus, Bacillus subtilis, Bacillus pumillus, E. coli, Klebsiella pneumoniae, Proteus vulgaris, Candida albicans and A. niger (Sini et al., 2011). Similarly, petroleum ether extract of C. zeylanica roots have inhibiting potential against S. aureus, Bacillus subtilis, Klebsiella pneumonia and Proteus vulgaris (Rengapriya et al., 2012).

The efficacy of plant extracts against bacterial growth might be due to phytoconstituents like phenolics and flavonoids present in plant extracts. So, antibacterial potential of Capparis extracts might be due to higher phenolic and flavonoid compounds found in different parts of Capparis species (Zia-ul-Haq et al., 2011; Imran et al., 2014). Proestos et al. (2006) studied the correlation between antimicrobial activity of plant extracts and phenolic acids. The researchers studied the phenolic acids composition by employing RP-HPLC with UV detection and further GC- MS was used for phenolic acids characterization. It was found that plant extracts with higher phenolic and flavonoid compounds exhibit good antibacterial activity. Capparis extracts are also rich in antioxidant compounds, phenolics, flavonoids, rutin, tocopherols, carotenoids and vitamin C (Tlili et al., 2011a,b; Imran et al., 2014).

The inhibited growth of E. coli and S. aureus can be correlated with flavonoid compounds in C. decidua extracts (Sharma and Kumar, 2009). Such studies show that the extracts rich in phenolic acids and flavonoids, exhibit better antimicrobial potential (Sivropoulou et al., 1995). Secondly, the difference in the antimicrobial potential of different solvents might be due to the difference in polarities and chemical characteristics of these solvents (Sultana et al., 2009).

Conclusion

Methanolic extracts of the selected parts of both species were more effective in inhibiting the growth of studied bacteria in comparison with other solvents. The results manifested that the bacterial population were maximally inhibited by C. decidua root extracts except B. subtilis while S. aureus was more sensitive to methanolic flower extract of C. decidua. Based on these findings, ex vivo confirmatory studies should be carried out for further explanation of the effectiveness of these species.

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Author:Gull, Tehseen; Sultana, Bushra; Bhatti, Ijaz Ahmad; Jamil, Amer
Publication:International Journal of Agriculture and Biology
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Date:Aug 31, 2015
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