Pure Appl. Biol., 7(1): 138-151, March, 2018 http://dx.doi.org/10.19045/bspab.2018.70017
Research Article
Phytochemical, antibacterial and antifungal activities of leaves and bark of Colebrookea oppositifolia: an ethnomedicinal plant
Muhammad Ajaib1*, Samia Abid1, Musfirah Anjum1, Qumqum Noshad1, Muhammad Faheem Siddiqui2 and Muhammad Asif Iqbal3 1. Department of Botany, Mirpur University of Science and Technology (MUST), Mirpur-10250 (AJK)- Pakistan 2. Department of Botany, University of Karachi, Karachi 75270-Pakistan 3. Department of Chemistry, University of Management and Technology (UMT), Lahore-Pakistan *Corresponding author’s email: [email protected] Citation Muhammad Ajaib, Samia Abid, Musfirah Anjum, Qumqum Noshad, Muhammad Faheem Siddiqui and Muhammad Asif Iqbal. Phytochemical, antibacterial and antifungal activities of leaves and bark of Colebrookea oppositifolia: an ethnomedicinal plant. Pure and Applied Biology. Vol. 7, Issue 1, pp138-151. http://dx.doi.org/10.19045/bspab.2018.70017 Received: 21/10/2017 Revised: 05/01/2018 Accepted: 05/01/2018 Online First: 17/01/2018 Abstract The phytochemical, antibacterial, antifungal and MIC (minimum inhibitory concentration) study of the leaves and bark of Colebrookea oppositifolia Smith was evaluated. Phytochemical screening C. oppositifolia Sm. indicated the presence of alkaloids, flavonoids, glycosides, steroids and saponins whereas terpenoids, tannins, and cardiac glycosides are showed only in bark extract. Petroleum ether extract showed maximum effectiveness against P. aeruginosa with bark extract (18±2.2 mm) and maximum antifungal potential exhibited by petroleum ether extract of leaves (15 ±0.3 mm) against A. nigar. It was noticed that the extracts in distilled water had not shown much potential. The MIC assay was carried out for further verification of results obtained by the Zone of Inhibition against both bacterial and fungal strains. The lowest MIC value is 0.09±0.3 at 1 mg/mL against E. coli exhibited by bark extract and 0.09±0.2 at 0.9 mg/mL against K. pneumoniae shown by leaf extract. Whereas, in fungi minimal MIC value exhibited 0.11±0.3 at 0.9mg/mL by leaf extract against A. oryzae. Hence, it is recommended further investigation to explore toxicological and clinical trials before using this plant as drug. Keywords: Antibacterial; Antifungal;Colebrookea oppositifolia; Ethnomedicinal plant Introduction properties by the identification of a critical Products obtained from plants have been number of helpful compounds [1, 2]. used medicinal purposes since pre-historic Ethno-botanical plants frequently use as time. For instance, certain conventional pharmaceuticals, Nutraceuticals, makeup pharmaceuticals including numerous trees, and nourishment supplements [3]. With an bushes, climbers and herbs have been expansion of resistant microorganisms, utilized to remedy of certain diseases. conventional plants are being researched Traditionally important plants have become for their antibacterial and therapeutic sources for examining their bioactive qualities. The antibacterial and antifungal impact of a plant is because of the
Published by Bolan Society for Pure and Applied Biology 138 Ajaib et al. phytochemicals such as saponins, phenolic The plant Colebrookeaoppositifolia Sm. compounds, tannins, terpernoids, heart commonly known as Bansa Siyah was glycosides, alkaloids and flavonoids [4]. collected from the Kakra town, District Numerous emphasis made on deciding the Mirpur AJK. Some plants of that species antimicrobial action of plant concentrates also seen in Khoirata, Peer Gali and Kotli. found in local societies in the form of oils, This plant belongs to the family Lamiaceae, flavonoids, alkaloids, lactones etc.[5]. also called as Mint family. The plant Foodborne diseases fall out 76 million species collected was mounted on individuals every year in the United States, herbarium sheet and submitted in the represent almost 325,000 hospitalizations Herbarium Department of Botany MUST, and more than 5,000 passings. The diseases Bhimber campus. A voucher No. MUST. caused by food are generally dangerous, even fatal for example, microscopic organisms, growths, infections and parasites that enter the body essentially by the ingestion of debased sustenance [6]. Because of the rapid resistance to antibiotics by using allopathic medicines, there is a need to grow new and inventive antimicrobial compounds. As plants are cheap, non-poisonous source, there is a long convention of utilizing them as a part of the treatment of irresistible illness in Cameroonian people[7]. There are numerous strategies for hindering or inactivating microorganisms by utilizing plant derived products as antimicrobials BOT. 880 to obtain for further reference. [8]. Figure 1. Colebrookea oppositifolia Colebrookea oppositifolia Smith (Figure1) Test organisms is a genus of family Lamiaceae/Labiateae. Total six bacteria were selected, the three C. oppositifolia is commonly known as strains were from gram Bansa.It is medicinally important plant and positiveEnterococcus it is utilized locally as sickness for faecalis,Staphylococcus aureus and differentpurposes. The leaves paste is Bacillus subtilis and two strains of gram utilized for wound healing and bruises negativeKlebsiellapneumonia,Escherichia while the paste of its roots is utilized against coli and Pseudomonas aeruginosaand two the epilepsy [9]. Juice assimilated by strains of fungal i.e. Aspergillus oryzae and beating young leaves is utilized as Aspergillusnigerwere selected used for anthelmintic [10]. The root and stem juice investigation of antimicrobial perspective are used in eye diseases. It has intriguing of C. oppositifolia Smith. Four standard antibacterial action and had exceptional antibiotics of bacterial strains radical searching potential to be utilized as (azithromycin, methicillin, amoxicillin and cell reinforcement [11]. Keeping in view ampicillin) and four standard antibiotics of the significance of ethno botanical status of fungal strains (nystatin, tezole, fungivin, C. oppositifoliaSmith and other members of and griseofulvin) were tested for comparing family Lamiaceae the present investigation the potential of plant extract with them. was carried out. Methodology adopted Materials and methods The whole plant was washed with cold Plant material water for removing of mud. After washing the plant parts such as bark and leaves were
139 Pure Appl. Biol., 7(1): 138-151, March, 2018 http://dx.doi.org/10.19045/bspab.2018.70017 separated and kept for drying under room Chloroform, Methanol and distilled water temperature about 20-30 days. After drying and then evaporated to get final the powder was made with the help of concentrated form of extracts.Thecorporeal pestle and mortar and then exposed for analysis of herbal extracts considered its maceration method [12]. texture, colour and appearance, and Maceration of the plant specimen calculated yield before further study was The shade dried plant material of C. accompanied. oppositifolia was subjected for maceration Yields percentage (%) was calculated with different solvents i.e. Petroleum ether, the following method:
Weight of plant extract 퐸푥푡푟푎푐푡𝑖표푛 푦𝑖푒푙푑 = × 100 푊푒𝑖푔ℎ푡 표푓 𝑖푛𝑖푡𝑖푎푙 푝푙푎푛푡 푠푎푚푝푙푒
Qualitative study of phytoconstituents Phytochemical recognition key The phytochemical investigation of plant − = Absent, + = Present in minimal extracts were done by using typical process quantity, ++ = Existing in moderate [13]. quantity whereas, +++ = Present in Estimation of antimicrobial activity noticeableamount. Six bacterial strains By using agar well diffusion method for the including three gram-positive antimicrobial activity of leaf and bark (Staphylococcus aureus, Bacillus subtilis, extracts of C. oppositifolia. For Enterococcus facealis) and three gram antibacterial activity methodology of [14] negative (Escherichia coli, Pseudomonas was used. For the bacterial strains i.e., P. aeruginosa, Klebsiella pneumoniae) were aeruginosa,K. pneumonia, E.faecalis, S. used for the estimation of antimicrobial aureus, B. subtilis and E.coli, the nutrient activity of bark and fruit of Cholebrookia agar media was used. For antifungal oppositifolia Smith. All of these bacteria activity the methodology of[15] was used. were pathogenic. Furthermore, strains of Potato Dextrose Agar media was used for fungal i.e.,A.oryzaeandA. niger, selected antifungal activity. for antifungal prospective. Assessment of minimum inhibitory We also used some standard discs, to find concentration (MIC) out the receptiveness of the microbial Plant extracts of C. oppositifolia for MIC strains. Which is used as the test organisms was investigated by broth-dilution method (Tables 3 & 4). According to these adapted by [16]. standards, the specimens were categorized Statistical Analysis as vulnerable, transitional, and resistant Allthe statistical analysis was done utilizing prone. Leaf and bark of Microsoft excel 2010. C.oppositifoliaproducediffrent Zone of Results and discussion Inhibition in response of bacterial strains To checked the antimicrobial activity of the was recorded in mm. Zone of Inhibition plant material i.e. Colebrookea was produced by the p. ether extract of leaf oppositifolia Smith, extracts were prepared. showed maximum results i.e. 18±0.6 mm Macerated and their % yield was intended against B. subtilis. Whereas, bark extract in before further analysis, to provide glance of p. ether showed maximum Zone of the economical perspective of the Inhibition i.e. 18±0.8 mm against S. completepractice.The phytochemical aureusand 18±2.2 mm against P. screening the plant extracts of C. aeruginosa. The leaf of C.oppositifolia oppositifolia Smith was also achieved. Smith had displayed activity under the Results showed the presence and absence of range of 0±0 mm to 18±0.6mm. Bark had secondary metabolites (Table 1 & 2). exhibited the potential under the range of 0±0 mm to 18±2.2mm.
140 Ajaib et al.
Table 1. Phytochemical evaluations of Bark extracts of C. oppositifolia Smith Presence / Absence of Phytochemical Constituents
Plant Cardiac Solvent Alkaloids Steroids Tannins Flavonoids Glycosides Terpenoids Saponens Part Glycosides P. Ether ++ _ _ _ _ + + +++
Chloroform + + _ ++ _ +++ + ++
Bark Methanol +++ ++ + _ + + _ _ Aqueous _ _ + + _ _ _ _
Table 2. Phytochemical evaluations of Leaves extract of C. oppositifolia Smith Presence / Absence of Phytochemical Constituents Plant Solvent Cardiac Alkaloids Steroids Tannins Flavonoids Glycosides Terpenoids Saponens Part Glycosides P. Ether +++ + _ + + _ _ ++ Chloroform + ++ _ _ + _ _ +
Leaves Methanol ++ + _ _ ++ _ _ +
Aqueous _ _ _ + _ _ _ _
Table 3. Zone of Inhibition (mm) produced by the bacterial strainsagainst standard antibiotic discs Zone of inhibition (mm) Antibiotic Conc S. standard disc (µg) E. coli P. aeruginosa K. pneumoniae E. faecalis B. subtilis aureus Methicillin 15 8±0.6 9±1.2 9±0.5 8±0.5 8±0.7 8±0.5 Amoxicillin 30 8±1.5 9±1.0 8±1.0 9±1.3 - 11±1.0 Azithromycin 16 14±2.6 14±1.3 10.5±0.7 11±0.5 10±1.5 12±1.5 Ampicillin 10 9±1.7 8±2.0 9±1.2 8±1.0 7±0.5 8±0.5 GeneralResponse Resistant Resistant Resistant Resistant Resistant Resistant
Table 4. Zone of Inhibition (mm) of fungal strains against the standard antifungal discs Concentration Zone of inhibition (mm) Antifungal standard disc (µg/ml) A.niger A.oryzae Nystatin 99.9 23±2.1 30±1.6 Tezole 99.9 23±0.6 67±2.6 Fungivin 99.9 16±3.5 40±1.8 General Response Intermediate Intermediate
141 Pure Appl. Biol., 7(1): 138-151, March, 2018 http://dx.doi.org/10.19045/bspab.2018.70017
The extract of leaf had showed supreme muchalike potential with little difference. potential in contrast to S. Aureusas The potential exhibited by both parts was compared to the bark with the petroleum better as compared to standard discs. ether and aqueous, as shown in (Figure 2). Theleast potency was showed by the Whereas, chloroform extracts of leaf and methanolic extracts of both parts (Table5). bark, mutually had displayed nearly
Zone of Inhibition (mm) against S. aureus 25
20
15 Leaf Bark 10 Azithromycin
5 Ampicillin ZoneofInhibition (mm) 0 P.E Chl. MeOH Aq. Solvents
Figure 2. Graphical representation of the Zone of Inhibition against S. aureus
Table 5. The Zone of Inhibition produced by both parts of C.oppositifolia against the bacterial strains was documented in mm
Zone of Inhibition (mm)
Plant Solvents S. aureus E. coli P. aeruginosa K. pneumoniae E. faecalis B. subtilis Part P. ether 15±1.6 14±0.5 14±1.2 14±1.0 16±2.0 18±0.6 Chloroform 13±0.8 12±0.5 12±0.5 14±0.7 14±1.5 12±0.3 Methanol 10±0.8 11±0.6 10±0.3 12±0.6 11±0.7 13±0.8 Leaf Aqueous 17±0.3 17±0.3 16±1.0 15±0.3 15±0.5 17±1 P. ether 18±0.8 15±0.3 18±2.2 17±2.0 13±0.4 15±0.3 Chloroform 11±0.8 10±0.3 10±0.3 13±0.5 12±0.4 11±0.6 Methanol 11±0.8 11±0.5 11±1.2 10±1.0 16±0.7 10±0.5 Bark Aqueous 16±0.5 17±0.3 15±2.8 11±0.6 15±0.6 17±0.3
Both leaf and bark extracts had showed The leaf extract had exhibited the record marvelous potential against E. coli as well potential against P. aeruginosaas well. with the extracts in distilled With the petroleum ether being the most waterdisplaying the record potential in effective one. The aqueous extracts of response of the specific bacterial strain bothparts of C. oppositifolia Smith had (Figure 3). The p. ether extracts of both showed also satisfactory results against the parts had showed also better results against particular bacterial strains (Figure 4). E. coli.
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Zone of Inhibition (mm) against E. coli
20 18 16 14 Leaf 12 10 Bark 8 Azithromycin 6 4 Ampicillin ZoneofInhibition (mm) 2 0 P.E Chl. MeOH Aq. Solvents Figure 3. Graphical representation of the Zone of Inhibition against E. coli
Zone of Inhibition (mm) against P. aeruginosa
25 )
20
15 Leaf
10 Bark
Azithromycin 5 Ampicillin
ZoneofInhibition (mm 0 P.E Chl. MeOH Aq. Solvents
Figure 4. Graphical representation of the Zone of Inhibition against P. aeruginosa
Petroleum ether extract of bark (17±2.0 satisfactory results against E. faecalisi.e. mm) and aqueous extract of leaf (15±0.3 17±1 mm and 17±0.3 mm. Whereas, mm) exhibited maximum potential against chloroform bark extract and methanolic K. pneumoniae. Petroleum ether leaf leaf extract shown minimal potential. The extract and chloroform leaf extract had potential showed by both parts was better as showed somewhat similar results against compared to antibiotic discs which were particular strains of bacteria i.e., 14±1.0 used as standard (Figure 6). mm and 14±0.7 mm respectively. The The leaf extract had showed incredible activity displayed by both parts was greater, activity against B. subtilis, as well with the as compared to antibiotic discs (Figure 5). extracts soaked in petroleum ether The petroleum ether extract of leaf (16±2.0 represented the maximum potential i.e., mm) and methanolic extract of bark 18±0.7 against the particular bacterial (16±0.7 mm) showed maximum potential strain. The aqueous extracts of both parts against E. faecalis. Aqueous extract of both had expressed significant activity against B. leaf and bark of C. oppositifolia Smith subtilis (Figure 7). exhibited somewhat similar and
143 Pure Appl. Biol., 7(1): 138-151, March, 2018 http://dx.doi.org/10.19045/bspab.2018.70017
Zone of Inhibition (mm) against K.pneumoniae 20 ) 15 Leaf 10 Bark 5 Azithromycin
0 Ampicillin P.E Chl. MeOH Aq. ZoneofInhibition (mm Solvents
Figure 5. Graphical representation of the Zone of Inhibition against K. pneumonia
Zone of Inhibition (mm) against E. faecalis 18 16 14 12 10 Leaf 8 Bark 6 4 Azithromycin 2 Ampicillin
ZoneofInhibition (mm) 0 P.E Chl. MeOH Aq.
Solvents
Figure 6. Graphical representation of the Zone of Inhibition against E. faecalis
Zone of Inhibition (mm) against B. subtilis
25 20 15 Leaf
10 Bark 5 Azithromycin 0 Ampicillin ZoneofInhibition (mm) P.E Chl. MeOH Aq. Solvents
Figure 7. Graphical representation of the Zone of Inhibition against B. subtilis MIC Assay showed maximum activity against E. coli Themethanolic extracts were only (Table 6). employed for the MIC estimation of C. Methanolic extract inhibited growth of S. oppositifolia Smith. The leaf extract aureus very effectively and MIC value was repressedthe growth of S. 0.49±0.01 at conc. of 0.9 mg/mL and aureousefficientlywhile the bark extract 0.10±0.5 at conc. of 0.8 mg/mL (Figure 8).
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The bark extract contained the significant of C.oppositifoliaSmith were very effective potential against the all microbes, so we can against E. coli, their MIC values are say the bark extracts were more effective 0.10±0.33 and 0.09±0.3 at conc. of 0.9 and than leaf extracts. The bark and leaf extracts 1 respectively (Figure 9).
Table 6. MIC values (mg/mL) exhibited by leaf and bark of C. oppositifolia against Gram- positive and Gram-negative bacterial strains. K. S. aureus E. coli P. aeruginosa E. faecalis B. subtilis Plant pneumoniae Part Conc. MIC Conc. MIC Conc. MIC Conc. MIC Conc. MIC Conc. MIC 0.10± 0.13±0. 0.09± Leaf 0.9 0.49±0.01 0.9 0.8 0.9 0.8 0.14±0.5 0.7 0.10±0.3 0.33 7 0.2 0.15±0. 0.10± Bark 0.8 0.10±0.5 1 0.09±0.3 0.9 0.8 0.8 0.12±0.7 0.8 0.11±0.3 5 0.3
MIC values against S. aureus 0.4 0.35 0.3 0.25 0.2 Leaf 0.15
MIC(mg/mL) 0.1 Bark
0.05
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1
Conc.
Figure 8. Graphical representation of the MIC values against S. aureus
MIC values against E. coli
0.6
0.5 0.4 0.3 Leaf 0.2
MIC(mg/ mL) Bark 0.1 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 Conc.
Figure 9. Graphical representation of the MIC values against E. coli
145 Pure Appl. Biol., 7(1): 138-151, March, 2018 http://dx.doi.org/10.19045/bspab.2018.70017
The bark extract displayedsupreme The bark extract showed maximum potential in response of P. aeruginosa. The potential against E. faecalis then leaf leaf extract were less effective than bark extract. The bark extracts were extra extract (Figure 10). MIC values estimated effective as compared to leaf extracts in that for both parts were 0.13±0.7 at 0.81 mg/mL case. MIC values estimated for both parts and 0.15±0.5 at 0.9 mg/mL against E. faecalis were 0.14±0.5 at 8.0 correspondingly. mg/mL and 0.12±0.7 at 8.0 mg/mL The K. pneumoniae was stated to be correspondingly (Figure 12). maximum delicate against the bark extract. The B. subtilis was stated to be the When all methanolic extracts of C. maximum resistant, against the methanolic oppositifoliaSmith during analysis were extract of bark. Whereas, leaf extracts were taken to consideration. The bark extract extrareal than bark extracts. MIC values were more active than leaf extracts assessed for both parts of C.oppositifolia (Figure11). Actual MIC values estimated Smith against B. subtilis were 0.10±00.3 at for both parts were 0.09±0.2 at 0.9 mg/mL 0.7mg/mL and 0.11±0.3 at 0.8mg/mL and 00.10±0.3 at 0.8 mg/mL subsequently (Figure 13). correspondingly.
MIC values against P. aeruginosa
0.5 0.4
0.3
0.2 Leaf
MIC(mg/mL) Bark 0.1
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 Conc.
Figure 10. Graphical representation of the MIC values against P. aeruginosa
MIC values against K. pneumoniae 0.5 0.4
0.3 0.2 Leaf
MIC(mg/mL) Bark 0.1 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 Conc.
Figure 11. Graphical representation of the MIC values against K. pneumoniae
146 Ajaib et al.
MIC values against E. facealis 0.5 0.4
0.3 0.2 Leaf
MIC(mg/mL) 0.1 Bark
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 Conc.
Figure 12. Graphical representation of the MIC values against E. faecalis
MIC values against B. subtilis
0.5 0.4
0.3 0.2 Leaf
MIC(mg/mL) Bark 0.1 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1 Conc.
Figure 13. Graphical representation of the MIC values against B. subtilis The antimycotic potential of the C. The A. oryzae strain had presented resistant oppositifolia Smith was estimated using towards the plant extracts in comparison to two fungal strains i.e. A.niger and A.oryzae. A. niger. The supreme potential against the The extracts had shown variable potential respective fungal strain was showed by the against these fungal strains with the distilled water extract of both parts. The extreme potency being showed by the leaf extract macerated in methanol had showed extracts soaked in distilled water. However, resistance compared to other extracts. The petroleum ether, chloroform and potential exhibited by the bark extract of C. methanolic extracts had showed minimal oppositifolia Smith in chloroform was potential against these fungal strains (Table greater compared to leaf extract (Figure 15 7). & 16). The A. niger was additional sensitive near The extracts macerated in the methanol the plant extracts when equated to the A. were also utilized for the MIC estimation. oryzae. The supreme potential against the The leaf extract had exhibited maximum A. niger had showed by the aqueous extract potential against A. niger with the MIC of leaf. The petroleum ether extract of leaf value 0.55±0.5 at 0.9mg/mL whereas, bark had displayed less potential as compared to extract showed 0.50±0.5 MIC value at the petroleum extract of bark against the A. 0.8mg/mL (Table 8). niger(Figure 14; Table 8).
147 Pure Appl. Biol., 7(1): 138-151, March, 2018 http://dx.doi.org/10.19045/bspab.2018.70017
Table 7. Zone of Inhibition exhibited by leaf and bark of C. oppositifolia Smith against fungal test organisms Zone of Inhibition Plant Part Solvents A.niger A.oryzae P. ether 10±0.3 15±0.3
Chloroform 14±0.6 16±0.6 Methanol 14±0.6 17±0.9 Leaf Aqueous 28±0.3 25±0.6 P. ether 14±0.3 15±1.2
Chloroform 15±0.9 15±0.7
Bark Methanol 16±0.9 20±1.2 Aqueous 24±0.6 24±1.5
Table 8. MIC values (mg/mL) exhibited by leaf and bark of C. oppositifoliaagainst fungal test organisms Plant A. niger A. oryzae Part Conc. MIC Conc. MIC Leaf 0.9 0.55±0.5 0.8 0.11±0.3 Bark 0.8 0.50±0.5 0.8 0.21±0.5
Zone of Inhibition (mm) against A. niger
35 30 25 20 Leaf
15 Bark 10 5 Fungivin 0 Tezole ZoneofInhibition (mm) P.E Chl. MeOH Aq. Solvents
Figure 14. Graphical representation of the Zone of Inhibition against A. niger
Zone of Inhibition (mm) against A. oryzae 80 60 Leaf 40 Bark 20 Fungivin 0 Tezole P.E Chl. MeOH Aq.
ZoneofInhibition (mm) Solvents
Figure 15. Graphical representation of the Zone of Inhibition against A. oryzae
148 Ajaib et al.
(a) (b) (c) (d)
(e) (g) (f) (h) Figure 16. Zone of inhibition produced by: (a) Aqueous leaf extract against A. niger(b) P. ether bark extract against B. subtilis(c) Aqueous leaf extract against A. oryzae (d) Aqueous leaf extract against E. coli (e) Aqueous leaf extract against E. faecalis(f)MeOH leaf extract against E. faecalis(g) Aqueous leaf extract against K. pneumonia (h) Aqueous bark extract againstE. faecalis
Discussion investigation, in spite of their fundamental The highest yield obtained from the leaf of resemblances stated by [19]. C. oppossitifolia Smith was (17.13%). The The tendency of the antibacterial action highest % extraction of yield of the leaf of shown by the extracts of C. oppositifolia C. oppossitifolia Smith was 7.73% which Smith was nearly similar to that recorded by was nearly similar to the yield extract [20] on water and methanolic extracts from recorded by [17] during the assessment of diffrent South African plants. The some Indian plants with their ethno determined potential was given by leaf botanical values for their microbial extract of plant.The bark extract had potential. The lowest yield got in leaf providing good to reasonable potential. extracts of C. oppossitifolia Smith was 2.6% However, the extracts of leaf and bark had in methanol which was in accord to the yield showed almost alike activity with minute extract gained from the plant (leaf) extract of differences. lamiaceae family i.e. Menthalongi folia (L.) Within solvents overall, the petroleum ether Huds. reported by [18] during screening of extracts of C. oppositifolia Smith had some selected medicinal important plants extraordinary antibacterial potential in extracts for In vitro antimicrobial activity. contradiction of all the organisms The phytochemical screening of C. oppositi experienced during investigation. The folia Smith showed the occurrence of maximum potential exhibited was 18±2.2 alkaloids, steroids, terpenoids, saponins, mm. Similar results were reported by [2] glycosides, tannins, reducing sugar, during analysis of antimicrobial and flavonoids and cardiac glycosides in antioxidant activities of Chenopodium sufficient amount. ambrosioides. This distinct activity had Two types of bacterial strains, i.e. gram provided the evidence that the non-polar positive and gram negative were used for fractions i.e.,petroleum ether were vigorous
149 Pure Appl. Biol., 7(1): 138-151, March, 2018 http://dx.doi.org/10.19045/bspab.2018.70017 in their activity compared to the relatively excellent results against selected bacterial polar fractions i.e., methanol by [21]. and fungal strains. The MIC assay had The further amplification near the further provided the antimicrobial potential assessment of the antibacterial potential of of plant’s extracts. Thus, it can be concluded C.oppositifolia Smith was providing by that all results are significant on the basis of adopting microbroth dilution assay to increasing the concentration of the extracts. estimate MIC value. And in this The antimicrobial activity is may be due to technique,methanolicextracts were only the presence of secondary metabolites i.e., subjected to the MIC assay, because of its alkaloids, flavonoids, tannins, sterols etc. potential for the steady extraction of Authors’ contributions constituents got antibacterial potential. The Conceived and designed the experiments: M MIC value of methanolic extracts of the Ajaib, Guide and wrote the article: S Abid, plant under observation range from 0.09±0.2 Performed the experiments: M Anjum, Q to 0.49±00.01 mg/mL. Noshad & MA Iqbal, Analyzed the data: M The MIC potential displayed by the S. Ajaib & MF Sidduqui. aureus was estimated to be ≥ 0.10±0.5 References mg/mL. Furthermore, the MIC activity 1. Kamran SH, Ahmad M, Shahwar D showed against E. coli was estimated to be ≥ &Ajaib M(2016). Anti-diabetic and anti- 0.09±0.3, against P. aeruginosa was ≥ oxidant status of Loranthus pulverulentus 0.13±0.7, against K. pneumoniae was ≥ obtained from two different hosts. Bangl 0.09±0.2, against E. faecalis was ≥ 0.12±0.7 J Pharmacol 11: 181-189. and ≥ 0.10±0.3 against B. subtilis. The MIC 2. Ajaib M, Hussain T, Farooq S &Ashiq M evaluations against the bacterial strains (2016). Analysis of Antimicrobial and provided the evidence that the bark extract Antioxidant Activities of was more effective against P. aeruginosa Chenopodiumambrosioides, An followed by S. aureus. This results because Ethnomedicinal Plant. J Chem 11. of the fact that ,the bark was additional 3. Sharma S, Kelly TK & Jones PA (2010). developed and ripe than leaves and may Epigenetics in cancer Carcinogenesis. comprise some pigments and other 31: 27-36. composites which have been documented to 4. Doherty VF, Olaniran OO & Kanife UC inhibit with the antimicrobial activity of (2010). Antimicrobial activities of extracts stated by [22]. Aframomum melegueta (Alligator MIC efficiency for the fungal strain of the A. pepper). Inter J Bio 2: 126-131. niger, was assessed to be within the range of 5. Ngwendson JN, Bedir E, Efange SMN, 0.50±0.5 to 0.55±0.5mg/mL, and A. oryzae Okunji CO, Iwu MM, Schuster BG & was valued to be within the range of Khan IA (2003). Constituents of 0.11±0.3 to 0.21±0.5mg/mL. So, it was Peucedanumzenkeri seeds and their clearly established that the potential of the antimicrobial effects, Die Pharmazie- plant against fungal strain was greater in A. Inter J PharmaceuSci58: 587-589. nigerthanA. oryzaeparallel findings 6. W.H.O. (2010). World Health observed by [22] during investigation of Organization, “World health statistics antimicrobial activity of 2010,” World Health Organization. TamarindusindicaLinn.and [23] screened 7. Djeussi DE, Noumedem JA, Seukep JA, the antimicrobial and antioxidant activities Fankam AG, Voukeng IK, Tankeo SB, ofCirsiumwallichiiDC. Nkuete AH & Kuete V (2013). Conclusion Antibacterial activities of selected edible On the basis of phytochemical and plants extracts against multidrug- antimicrobial screening of Colebrookea resistant Gram-negative bacteria. BMC oppositifolia Smith, the results showed that complem alter med13: 164. the leaf and bark extracts had provided
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