VOL.

VIII, ISSUE XXVII, OCT 2018 MULTILOGIC IN SCIENCE ISSN 2277-7601

An International Refereed, Peer Reviewed & Indexed Quarterly Journal in Science, Agriculture & Engineering
SCREENING SOME LOCAL PLANTS FOR ANTIBACTERIAL ACTIVITY AGAINST GRAM POSITIVE ORGANISMS
R. B. Gade1*, A. A. Deshmukh2, M. K. Patil3, A. V. Bhonsle4, N. Z. Gaikwad5, M. A. Khan6, P.V. Jadhav7
College of Veterinary and Animal Sciences, Udgir (M.S.)
Maharashtra Animal & Fishery Sciences University, Nagpur.
(Received: 10.08.2018; Revised: 19.08.2018; Accepted: 23.08.2018)
(RESEARCH PAPER IN VETERINARY AND ANIMAL SCIENCES)
Abstract
The aim of the study was to evaluate the antibacterial effect of aqueous and methanol leaf extracts of Aegle marmelos, Bauhinia racemosa and
Psidium gaujava plant extracts against Gram Positive organisms namely Staphylococcus aureus and Streptococcus pyogenes. The aqueous
and methanol leaf extracts were prepared by the maceration method. The antibacterial activity of the aqueous and methanol leaf extracts of
different concentrations ( 5mg, 10mg, 15mg, and 20 mg) and standard antibacterial drug Cefuroxime(30µg) were studied by the dics diffusion
method. The antibacterial efficacy of aqueous and methanol leaf extracts were compared with the Cefuroxime(30µg). The results revealed
that both the aqueous and methanol leaf extracts of Aegle marmelos, Bauhinia racemosa and Psidium gaujava possess antibacterial activity
against both Staphylococcus aureus and Streptococcus pyogenes . The aqueous and methanol leaf extracts of Psidium gaujava has better
antibacterial activity against Staphylococcus aureus and Streptococcus as compared to aqueous and methanol leaf extracts of Aegle marmelos
and Bauhinia racemosa. The antibacterial activity of aqueous and methanol leaf extracts of all plants under study was lesser than that of
Cefuroxime(30µg).
Key wards: Aegle marmelos, Bauhinia racemosa, Psidium gaujava, Antibacterial activity, Cefuroxime.
Introduction The stock solutions of aqueous and methanol leaf extracts were
Despite of the substantial advances of the biomedical sciences and prepared by dissolving 5 mg, 10 mg, 15 mg, and 20 mg of respective
public health polices in the past century, the world has witnessed an extracts in 250 µL of DMSO. The sterile disc of 6 mm diameter were
expansion of the emerging and re-emerging infectious diseases. On the saturated with stock solution to obtain various concentrations (5, 10,
contrary there had been a drastic decrease in the number of 15, 20 mg/disc) of the extracts. The DMSO loaded discs were also
antimicrobial drugs launched to the market by the pharmaceutical prepared and were used as negative control. The discs were air dried
companies, which blossomed the interest worldwide to bring for 3 to 4 hr to evaporate the solvent completely before placing them
medicinal plants as an alternative to the synthetic antimicrobial drugs on agar plates. The discs of Cefuroxime (30µg) were used as a positive
(Araujo et al. 2014).The ever problem of development of bacterial control.
resistance against the synthetic antimicrobial compounds also paves Disc Diffusion Method
the way to develop antimicrobial drugs of plant origin. The present The test for susceptibility of bacteria was performed by disc diffusion
study is therefore aimed for the search of medicinal plants which has method (Balouiri et al. 2016) Mueller-Hinton agar plates were poured
antibacterial properties and could serve as an alternative to the with a suspension of bacteria so as to obtain a uniform thick lawn
allopathic antimicrobial compounds. After going through the vast growth. The prepared extract discs and standard antibiotic discs were
literature for search of the new plants having the antimicrobial activity, kept in plates with sterile forcep at equal distance. The plates were
the plants namely Aegle marmelos, Bauhinia racemosa and Psidium incubated at 370C for 24 hr during which activity was evidenced by the
guajava were found to possess the antimicrobial properties. Therefore, presence of a zone of inhibition surrounding the disc.
the present study was designed to study the antibacterial properties of Statistical analysis:
the said plants against the mastitis causing organisms namely The data of the present investigation were statistically analyzed by
Staphylococcus aureus and Streptococcus pyogenes. applying SPSS Software.
Materials and Methods Results and Discussion
Identification, Collection and authentication of plant material In the present study aqueous and methanol leaf extracts (5mg, 10mg,
The fresh leaves of the plants Aegle marmelos, Bauhinia racemosa and 15mg and 20mg) of Aegle marmelos, Bauhinia racemosa and Psidiun
Psidium gaujava were collected from campus of college of Veterinary gaujava were evaluated for antimicrobial activity against S.aureus and
and Animal Science, Udgir and suburban areas of Udgir city. The plant S.pyogenes by using disc diffusion method at concentration of 5mg,
Aegle marmelos, Bauhinia racemosa and Psidium gaujava were 10mg, 15mg and 20mg. Table 1 shows the zone of inhibitions of
identified and authenticated by the botanist from the Department of aqueous and methanol leaf extracts of Aegle marmelos, Bauhinia
Botany, Shivaji Mahavidhyalaya, Udgir. racemosa and Psidium guajava against S.aureus and S. pyogenesat
Preparation of extract concentrations of 5mg, 10mg, 15mg and 20mg.
Fifty grams of the leaf powder of respective plants was taken in six The mean zone of inhibitions observed at 5mg, 10mg, 15mg and 20mg
separate 500 ml conical flasks. The 200 ml of distilled water and concentrations of aqueous leaf extract of Aegle marmelos were
methanol were respectively added to these six flasks. Flasks were 7.06±0.29mm, 8.10.±0.30mm, 9.13±0.37mm and 10.06±0.38 mm
Stoppard tightly and were kept in refrigerator for maceration for 48 hrs. respectively against S. aureus. The mean zone of inhibitions against S.
The flasks were shaken intermittently during the maceration period. pyogenes obtained by the aqueous leaf extract of Aegle marmelos were
After maceration, the contents in flask were filtered through the muslin 7.03±0.41mm, 8.20±0.45mm, 9.13±0.32mm and 11.20±0.28mm at
cloth and the filtrate so obtained was once again filtered through 5mg,10mg,15mg and 20mg respectively. The Cefuroxime (30µg)
Whatman filter paper no.1. This filtrate was then transferred to produced mean zone of inhibition as 26.03±0.10mm
sterilized evaporating bowl, which was then placed under fan for and22.23±0.35mm against S. aureus and against S.pyogenes
evaporation of the solvent so as to make the extract as much dry as respectively. The results indicate that the antibacterial activity of
possible. aqueous leaf extract of Aegle marmelos against S. aureus is dose
Bacterial strains dependant. The mean zone of inhibitions of methanol leaf extract of
The standard strains of gram positive bacteria namely Aegle marmelos at 5mg, 10mg, 15mg and 20mg were 6.10±0.05mm,
Staphaylococcus aureus (ATCC-43300) and Streptococcus pyogenes 7.06±0.14mm, 7.06±0.14mm and 8.10±0.11mm respectively against
(ATCC-12348) were selected for the study. S.aureus. The mean zone of inhibitions against S. pyogenes produced
Preparation of discs: by the methanol leaf extract of Aegle marmelos were 6.10±0.05mm,
7.06.±0.14mm, 8.10±0.11mm and 9.1±0.26mm at 5mg, 10mg, 15mg
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 VOL. VIII, ISSUE XXVII, OCT 2018 MULTILOGIC IN SCIENCE ISSN 2277-7601
An International Refereed, Peer Reviewed & Indexed Quarterly Journal in Science, Agriculture & Engineering
and 20mg respectively. The mean zone of inhibition produced by 10mg, 15mg and 20mg concentrations of methanol leaf extract of
Cefuroxime (30µg) against S.aureus and S.pyogenes were Psidium gaujava were 12.20±0.10mm, 12.20.±0.05mm,
22.16±0.24mm and 21.45±0.05mm . The mean zone of inhibitions 13.10±0.10mm and 15.20±0.10mm respectively against S. aureus.
against S. aureus observed at 5mg, 10mg, 15mg and 20mg However, the standard drug Cefuroxime is shown to exert more
concentrations of aqueous leaf extract of Bauhinia racemosa were pronounced zone of inhibition (28.00±0.50mm) against S. aureus.
6.00±0.20mm, 7.16.±0.20mm, 7.16±0.29mm and 8.23±0.17mm Similarly the zone of inhibition produced by the Cefuroxime (30ug)
respectively. While the mean zone of inhibitions of aqueous leaf was found to be higher (28.00±0.10mm) against S. pyogenes when
extracts of Bauhinia racemosa against S. pyogenes were compared to 5mg,10mg,15mg and 20mg concentrations of methanol
5.96±0.27mm, 7.20±0.30mm, 8.06±0.29mm and 9.20±0.17 mm at leaf extract of Psidium gaujava. The zone of inhibitions against S.
concentration of 5mg, 10mg, 15mg and 20mg respectively. The zone pyogenes exerted by the methanol leaf extract of Psidium gaujava were
of inhibition produced by the Cefuroxime were 20.00±0.57mm and 13.23±0.150mm, 14.36±0.350mm, 15.16±0.100mm and
19.70±0.90mm against S.aureus and S. pyogenes respectively. The 17.03±0.100mm at 5mg, 10mg, 15mg and 20mg respectively.
mean zone of inhibition produced by the methanol leaf extract of From study it is observed that all plants under study viz. Aegle
Bauhinia racemosa at 5mg, 10mg, 15mg and 20mg concentrations marmelos, Bauhinia racemosa and Psidium gaujava possess varied
against S. aureus were 7.10±0.14mm, 8.13.±0.14mm, 10.06±0.43mm level of antibacterial activity. From the table 1 it is also evident that the
and 11.16±0.14mm respectively. While, the standard drug Cefuroxime highest zone of inhibitions of 14.13±0.12mm and 15.20±0.17mm are
(30µg) produced maximum mean zone of inhibition (20.00±0.57mm) produced by aqueous leaf and methanol leaf extractsof Psidium
against S.aureus. The mean zone of inhibitions obtained by methanol gaujava respectively against S.aureus at 20mg. While,the zone of
leaf extract of Bauhinia racemosa against S. pyogenes were inhibitions of 16.13±0.12mm and 17.03±0.37 mm are produced by
6.26±0.35mm, 8.03.±0.18mm, 8.86±0.33mm and 11.00±0.57mm at aqueous leaf and methanol leaf extractsof Psidium gaujava at 20mg
5mg,10mg,15mg and 20mg respectively. The mean zone of inhibition respectively against S. pyogenes .
produced by the Cefuroxime (30ug) was found to be maximum Conclusion
(19.70±0.30mm) against S. pyogenes. The mean zone of inhibitions From above study it is observed that all plants under study viz. Aegle
against S. aureus observed at 5mg, 10mg, 15mg and 20mg marmelos, Bauhinia racemosa and Psidium gaujava possess dose
concentrations of aqueous leaf extract of Psidium gaujava were, dependant and varied level of antibacterial activity against S.aureus
11.50±0.57mm, 12.20±0.28mm, 13.20±0.20mm and 14.13±0.12mm and S. pyogenes. The aqueous and methanol leaf extracts of Psidium
respectively. While, the Cefuroxime (30µg) exhibited significantly gaujava comparatively shows better antibacterial activity against S.
higher zone of inhibition (24.30±0.57mm) against S. aureus in aureus and S. pyogenes as compared to aqueous and methanol leaf
comparison to all concentrations of aqueous leaf extract of Psidium extracts of Aegle marmelos and Bauhinia racemosa
gaujava. The zone of inhibitions obtained by the aqueous leaf extract References
of Psidium gaujava against S. pyogenes were 12.16±0.17mm, Araujo, A. A. D., L. A. L. Soares, M. R. A. Ferreira, M. A. D. S.
14.30±0.41mm, 15.20±0.15mm and 16.13±0.12mm at Neto, G. R. D. Silva, R. F. D. Araujo, G. C. B. Guerra, M. C. N. D.
5mg,10mg,15mg and 20mg respectively. While, the standard drug Melo. (2014). Quantification of polyphenols and evaluation of
Cefuroxime (30µg) produced the zone of inhibition (23.00±0.57) antimicrobial, analgesic and anti-inflammatory activities of aqueous
against S. pyogenes which supersedes the observed zone of inhibitions and acetone–water extracts of Libidibia ferrea, Parapiptadeni arigida
exhibited by aqueous leaf extract of Psidium gaujava at 5mg, 10mg, and Psidium guajava. Journal of Ethnopharmacology.156: 88–96.
15mg and 20mg concentrations. From these it is evident there is dose Balouiri M., M. Sadiki and S. K. Ibnsouda (2016) Methods for in
dependant antibacterial activity of aqueous leaf extract of Psidium vitro evaluating antimicrobial activity: A Review. Journal of
gaujava against S. aureus. The zone of inhibitions observed at 5mg, Pharmaceutical Analysis. 6: 71-79.

Table 1: Zone of inhibitions of aqueous and methanol leaf extracts of Aegle marmelos, Bauhinia racemosa and Psidium guajava
against S. aureus and S. pyogenes
Plant Zone of inhibition in mm (Mean ± SE)
extrac Concentrations
ts 5mg 10mg 15mg 20mg Cefuroxime(30µg)
S. aureus S. S. aureus S. S. aureus S. S. aureus S. S. S.
pyogenes pyogenes pyogenes pyogenes aureus pyogenes
AAM 7.06d±0.29 7.03d±0. 8.10c±0.30 8.20c±0.4 9.13c±0.37 9.13c±0.3 10.06b±0.3 11.20b±0. 26.03a±0. 22.23a±0.
** 41 ** 5 ** 2 8** 28 10 35
MAM 6.10d±0.05 6.10e±0.0 7.06c±0.14 7.06d±0.1 7.06c±0.14 8.10c±0.1 8.10b±0.11 9.10b±0.2 22.16a±0. 21.26a±0.
* 5 * 4 ** 1 ** 6 24 18
ABR 6.10d±0.05 6.10e±0.0 7.06c±0.14 7.06d±0.1 7.06c±0.14 8.10c±0.1 8.10b±0.11 9.10b±0.2 22.16a±0. 21.26a±0.
* 5 * 4 ** 1 ** 6 24 18
MBR 7.10e±0.14 6.26e±0.3 8.13d±0.14 8.03d±0.1 10.06c±0.4 8.86c±0.3 11.16b±0.1 11.00b±0. 20.00a±0. 19.70a±0.
5 8 3 3 4 57 57 30
APG 11.50e±0.5 12.16e±0. 12.20d±0.2 14.30d±0. 13.20C±0.2 15.20c±0. 14.13b±0.1 16.13b±0. 24.30a±0. 23.00a±0.
7** 17 8** 41 0** 15 2** 12 57 57
MPG 12.20d±0.2 13.45e±0. 12.20d±0.1 14.35d±0. 13.10c±0.1 15.16c±0. 15.20b±0.1 17.03b±0. 28.00a±0. 28.10a±0.
0** 23 5** 34 1** 14 7** 37 57 57
Note:* Significant (P<0.05) ** Highly Significant (P<0.01)Means with similar subscript are not significantly different
Where, AMA= Aqueous Aegle marmelos extract MAM= Methanol Aegle marmelos extract
ABR= AqueousBauhinia racemosa extract MBR= Methanol Bauhinia racemosaextract
APG= AqueousPsidium gaujava extract MPG= Methanol Psidium gaujava extract

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