Microbiological

Communication

Biosci. Biotech. Res. Comm. 10(4): 752-763 (2017)

Pharmacological activity of different solvent extracts

Tribulus terrestris

against multi drug resistant

Staphylococcus aureus

isolated from post-operative

wound patients

M.S. Uma Maheshwari, R. Rajendran and S. Vijayalakshmi

Associate Professor, Department of Microbiology, Tiruppur Kumaran College for Women, Tiruppur, India

Associate Professor, Department of Microbiology, PSG College of Arts and Science, Coimbatore, India

Research Scholar, PG and Research Department of Zoology, Division of Microbial Technology, Chikkanna

Govt. Arts College, Tirupur, India

ABSTRACT

Despite advances in infection control surgical site infections formerly called surgical wound infections, remain a substantial cause of

morbidity and mortality among hospitalized patients. The post-operative surgical site infection is the third most commonly reported

nosocomial infections, accounting for a quarter of all such infections. The early stages of invasive infection caused by Staphylococcus

aureus, also play major role in this process. In this present study a total of 50 pus samples were collected from patients having postop-

erative wound infections from the different surgical departments in the PSG Institute of Medical Sciences and Reasearch (PSG IMSR),

Coimbatore during Dec 2016 to July 2017. Identi cation of bacterial isolates was done by standard microbiological techniques. Further,

the antimicrobial susceptibilities were done against following antibiotics, Penicillin (10 units), Chloramphenicol (30mcg), Vancomycin

(30mcg), Streptomycin (10mcg), Neomycin (30mcg), Ampicillin (10mcg), Amoxicillin (10mcg), Amikacin (10mcg), Gentamycin (10mcg)

and Cipro oxacin (10mcg). Among these strains 100%, 30%, 90%, 50%, 40%, 100%, 50%, 20%, 30% and 60% were found to be exhibit-

ing a signi cant degree of resistance to antibiotics tested. Currently, the development of bacterial resistance has necessitated the search

for new antibacterial agents to combat the infectious disease using bio-medically active different solvent extract of Tribulus terrestris

for antimicrobial activity. In this present study the maximum inhibition was observed in Ethyl acetate, moderate inhibition in Ethanol,

minimum inhibition in Acetone, Chloroform, Methanol and no inhibition in aqueous and Petroleum ether. The results con rm that, the

plant appears to contain substances that exhibit broad antimicrobial activity against wound pathogens.

KEY WORDS:

STAPHYLOCOCCUS AUREUS, ANTIBIOTIC RESISTANCE, ANTIMICROBIAL ACTIVITY OF TRIBULUS TERRESTRIS

, POSTOPERATIVE

WOUND INFECTION

752

ARTICLE INFORMATION:

*Corresponding Author: maheswariuma938@gmail.com

Received 27

Oct, 2017

Accepted after revision 27

Dec, 2017

BBRC Print ISSN: 0974-6455

Online ISSN: 2321-4007 CODEN: USA BBRCBA

Thomson Reuters ISI ESC and Crossref Indexed Journal

NAAS Journal Score 2017: 4.31 Cosmos IF: 4.006

reserved.

Online Contents Available at:

http//www.bbrc.in/

DOI: 10.21786/bbrc/10.4/22

Uma Maheshwari, Rajendran and Vijayalakshmi

INTRODUCTION

Recurrently, the post operative hospital acquired infec-

tions constitutes a major problem in surgical patients

contributing to morbidity, mortality and increased

resource utilization and health care costs. Patients in

whose surgical site infection develop have an increased

number of associated complications, the high risk of

requiring a stay in ICU have two to three times higher

risk of mortality. Their hospital stay is increased by 7 to

12 days and they are  ve times more likely to require

readmission.The post-operative wound infections can

be caused by different groups of microorganisms like

bacteria, fungi and protozoa. However, different kind of

microorganisms can exist in polymicrobial communi-

ties, especially in the margins of wounds and in chronic

wounds (Percevil and Bowler, 2004, Anaya and Del-

linger, 2006 and Jain et al. 2014) .

The risk of invasive burn wound infection is in u-

enced by the extent and depth of the burn injury, vari-

ous host factors, and the quantity and virulence of the

microbial  ora colonizing the wound (Church et al.

2006). The common burn wound pathogens are Staphy-

lococcus aureus, Pseudomonas aeruginosa, Escherichia

coli and Klebsiella spp., which produce a number of

virulence factors that are important in the pathogenesis

of invasive infection (Tredget et al. 2004) are frequently

associated with post-operative wound infection. One of

the major problems worldwide is the increase in anti-

biotic-resistant strains of bacteria, mainly in hospitals,

that poses constrain for their control without consider-

able resources and expenditure (Ayliffe et al. 2000). It

has been well documented that most of the clinical iso-

lates of Staphylococcus aureus are multiple-drug resist-

ant currently used antibiotics. The problem of microbial

resistance is increasingly alarming and the outlook for

the use of antimicrobial drugs in the future is still uncer-

tain (Nascimento et al. 2000 Naaz 2017).

Due to this treatment of postoperative wound infec-

tion with antibiotics is becoming a challenge for the sur-

geon as multidrug resistance is reported to be high. It is

therefore, important to have knowledge regarding the

prevalent microorganism in the surgical units and their

susceptibility patterns to antibiotics so that proper treat-

ment can be started earlier. It is essential to take appro-

priate steps to curtail the spread of infection within

the unit (Tahir, 1995). One of the measures to combat

this increasing rate of resistance is to have continuous

investigations into new, safe and effective antimicrobi-

als as alternative agents to substitute with less effective

ones. Plants have been traditionally proved to be a rich

source of novel drug compounds, as the herbal mix-

tures have made large contributions to human health

and well-being (Ergin and Mutlu, 1999). A wide vari-

ety of secondary metabolites, such as tannins, terpe-

noids, alkaloids, quinones and  avonoids are endowed

with antimicrobial properties (Lewis and Ausubel, 2006

Mohammad et al 2015).

Currently, the research is being carried out to inves-

tigate ethno-botanical uses of plants prevailing among

native people (Sibanda and Okoh, 2007). There are

numerous reports evidencing the antibacterial activity

of plants against microorganisms (Sundharameshwari

and Radhika, 2007). Thus, it is very much necessary

to analyze the potential of the plants in combating the

antibiotic resistant organisms, (Al Maofari 2013).

Recurrently, the Tribulus terrestris is a strong herbal

remedy which is used for various purposes in folk and

modern medicine and sport, as well. It has been used

as a tonic, aphrodisiac, astringent, analgesic, stomachic,

anti-hypertensive, antibacterial, antifungal, skin infec-

tion and urinary anti-septic (Al-Bayati and Al-Mola,

2008). T. terrestris has been commonly used as a diuretic

as well as treatment for hypertension, hypercholester-

olemia and colic pains. (Wang et al., 1990).The leaves

of Tribulus terrestris are used traditionally for the treat-

ment of various kinds of wound. T. terrestris is found to

be a rich source of calcium (Bourke et al., 1992). Extract

from T. terrestris has immune stimulatory and antimi-

crobial effect (Sengul et al., 2009, Al Maofari 2013 )

against pathogens. Recently Naz et al., (2017) stated that

the leaf extracts of plants with a history of traditional

use should be tested using modern methods for activi-

ties against multidrug resistant human pathogens, with

the aim of discovering potential new drugs. So, hence

the present study has made an attempt to point out the

different solvent extraction of Tribulus terrestris against

multidrug resistant Staphylococcus aureus isolated from

wound of post-operative patients.

MATERIALS AND METHODS

A total of 50 pus samples was collected from patients

aseptically with a sterile cotton swab suffering from post

operative wound infection at the PSG Institute of Medi-

cal Sciences and Reasearch (PSG IMSR), Coimbatore

for the period of Dec 2016 – July 2017. For collection,

the wound sample was washed thoroughly with normal

saline and it was placed in the ice box. After reaching to

the laboratory of PG and Research Department of Micro-

biology, PSG College of Arts and Science, Coimbatore.

Finally the samples were incubated at 37ºC for 24 hours

in isolation of wound pathogens.

All the strains isolated from wound samples were seri-

ally diluted, plated onto Mannitol Salt Agar, it was incu-

bated at 37°C for 48hours. The colonies with characteristic

growth were subjected to routine biochemical test accord-

ing to the Bergey’s manual of systematic bacteriology.

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS PHARMACOLOGICAL ACTIVITY OF DIFFERENT SOLVENT EXTRACTS OF

TRIBULUS TERRESTRIS

753

Uma Maheshwari, Rajendran and Vijayalakshmi

The standard Kirby Bauer disk diffusion method was

used to determine the antimicrobial pro le of wound

isolates against 10 antimicrobial agents such as Peni-

cillin (10 units), Chloramphenicol (30mcg), Vancomy-

cin (30mcg), Streptomycin (10mcg), Neomycin (30mcg),

Ampicillin (10mcg), Amoxycillin (10mcg), Amika-

cin (10mcg), Gentamycin (10mcg) and Cipro oxacin

(10mcg). The diameters of the inhibition zone were

measured using a ruler under a colony counter appara-

tus. The results were expressed as sensitive (S), margin-

ally susceptible (I), and resistant (R).

The plants of Tribulus terrestris were collected from

the dry lands of Coimbatore and Tirupur regions and

identi ed. The identi cation was authenticated by the

Botanical Survey of India ,Tamilnadu Agricultural Uni-

versity Campus (TNAU),Coimbatore. The plant extracts

were prepared as per standard procedures. The leaf of T.

Terrestris plant (Fig. 1) were washed with sterile distilled

water to remove dirt, dried under shade and were ground

to powder using household electric blender. The 20g of

dry powdered Terrestris was weighed and transferred to

a conical  ask containing 100 ml of 80% of Ethanol,

methanol, Chloroform, acetone, Ethyl acetate and Petro-

leum ether (Fig. 2) respectively and allowed to soak at

ambient temperature for 72 hours. The extract was than

 ltered using Whatman no 1  lter paper and the  ltrates

were concentrated in vaccum at 40°C using a rotary

evaporator. Residues of the extracts made into suspen-

sions using sterile distilled water and sterile dimethyl

sulphoxide at a concentration of 500 mg/ml of Tribulus

extracts respectively.

The leaf of the puri ed Tribulus extract was tested

for antibacterial activity by standard agar well diffusion

method against pathogenic bacteria S.aureus. The pure

culture of bacterial pathogen was sub cultured on nutri-

ent agar. 20ml of nutrient agar were poured into petri-

plates. The well of 6mm diameter were made on nutrient

agar using gel puncture 100μl of fresh over night grown

culture of the respective bacteria were spread on nutri-

ent agar medium containing petriplates. The culture was

FIGURE 1. Tribulus terrestris

FIGURE 2. Extraction of Solvent from bioactive leaf

of Tribulus terrestris a) Petroleum ether, b) Acetone, c)

Ethanol, d) Aqueous, e) Ethyl acetate, f) Methanol, g)

Chloroform

FIGURE 3. Colony of Staphylo-

coccus aureus on Mannitol Salt

agar

754 PHARMACOLOGICAL ACTIVITY OF DIFFERENT SOLVENT EXTRACTS OF

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Uma Maheshwari, Rajendran and Vijayalakshmi

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS PHARMACOLOGICAL ACTIVITY OF DIFFERENT SOLVENT EXTRACTS OF

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755

swabbed uniformly using a sterile cotton swab and then

50μl of the plant extract solution was loaded into the

well. After incubation at 37ºc for 24 hours the different

zone of inhibition was measured.

Minimum inhibition concentration was made by the

lowest cost of the extract of T. Terrestris leaves where

it can show the bactericidal and bacteriostatic effect.

The test was performed in 96 well micro titer plates.

Microtiter plate wells from each column in row 1 were

marked and 100μl (500mg/ml) of stock (aqueous and

solvent extract) was added. 50μl of sterile distilled water

was added to rows 2-12. Two fold serial dilutions were

performed by transferring 50μl of solution from row 1

to 2, using a multichannel pipette. This was repeated

down the row 2 to 12. 40μl of double strength nutri-

ent broth and 10μl of bacterial culture was added to all

the wells in a separate column, so the  nal concentra-

tion of the inoculum in all the wells. Finally, to prevent

dehydration, the plates were covered with a plastic cover

and then incubated at 37ºC for overnight. The bacterial

growth was determined after adding of 40μl of 2, 3, 5

Tri Phenyl Tetrazolium Chloride Red (0.02mg/ml). The

Minimum inhibitory concentration (MIC) of the isolates

was taken as the lowest concentration of the antibiotic

of which the bacteria tested did not show visible growth

(Table: 1).

RESULTS AND DISCUSSION

Totally 50 clinical wound samples, 10 positive Staphylo-

coccus aureus were isolated from different wound sites

of patients admitted in surgical ward in the PSG Insti-

tute of Medical Sciences and Research (PSG IMSR) (Fig.

2), Coimbatore. All the wound samples were tested on

Mannitol Salt Agar (MSA) for the isolation of S.aureus.

On MSA, S.aureus colonies were appeared yellow colour

which is containing mannitol to detect mannitol fermen-

tation. In the routine microbiological laboratory, prompt

identi cation of the S. aureus was done by grams stain-

ing, catalase, oxidase, coagulase, IMViC, nitrate reduc-

tion test, triple sugar iron agar test. After performing, all

these tests was con rmed as S.aureus.

All the 10 S.aureus isolates were tested invitro to

determine their antibiotic susceptibility patterns by

antibiotic disc diffusion method and the following anti-

biotic discs were used for this assay are Penicillin (10

units), Chloramphenicol (30mcg), Vancomycin (30mcg),

Streptomycin (10mcg), Neomycin (30mcg), Ampicillin

(10mcg), Amoxycillin (10mcg), Amikacin (10mcg), Gen-

tamycin (10mcg) and Cipro oxacin (10mcg).. Totally 10

antibiotic discs were used to identify vancomycin resist-

ant Staphylococcus aureus. All the isolates were shown

multiple antibiotic resistances to the antibiotic tested

none of the isolates showed 100% resistant to antibi-

otics tested. The maximum resistant pattern percentage

(80%) was recorded in strain no MBUM02, MBUM06 and

MBUM07 followed by minimum resistant pattern per-

centage (30%) was recorded in strain no MBUM01 and

MBUM08.

Among 10 strains, 2 isolates were shown 30% resist-

ant against all the antibiotics tested with strain No.

MBUM01 and MBUM08 both were showed equally with

the following antibiogram: VAN- AMP-P followed by

the strain no. MBUM10, MTUM04 were showed 40%

with antibiogram of STR-CHL-AMP-P, STR-CHL-AMP-

CIP. Two strains MBUM03 and MTUM05 were showed

50% against the antibiotics tested with antibiogram

of VAN-AMX-NEO-AMP-P followed by one strain

no. MBUM09 was exhibit 70% resistant to antibiotics

tested: VAN-AMX-AMP-P-GEN-AK-CIP, these antibio-

gram was recorded in the isolate. Finally, three strains

MBUM02, MBUM06 and MBUM07 was showed the

highest resistant percentage of 80% with the antibio-

Table 1. MIC of different strains of Petroleum ether extract of Tribulus terrestris by broth

dilution method

Test

microorganism

Plant Extract (Petroleum ether)

Concentration (500mg/ml)

100

(μl)

12.5

(μl)

6.25

(μl)

3.12

(μl)

1.56

(μl)

0.78

(μl)

0.39

(μl)

0.19

(μl)

0.09

(μl)

0.04

(μl)

MTUM02 - - + + + + + + + + + +

MTUM03 - - + + + + + + + + + +

MTUM05 - - + + + + + + + + + +

MTUM06 - - + + + + + + + + + +

MTUM07 - - + + + + + + + + + +

MTUM09 - - + + + + + + + + + +

(-) indicates absence of growth and (+) indicates presence of growth of microorganisms.

Uma Maheshwari, Rajendran and Vijayalakshmi

756 PHARMACOLOGICAL ACTIVITY OF DIFFERENT SOLVENT EXTRACTS OF

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FIGURE 4. Antimicrobial activity of different solvent extract of Tribulus terrestris against

Staphylococcus aureus shows that the maximum inhibition was seen in Ethyl acetate extract of

Tribulus terrestris. The moderate inhibition was seen in Ethanol extract of Tribulus terrestris.

The minimum inhibition was seen in the Acetone, Chloroform extract Tribulus terrestris. No

inhibition of Tribulus terrestris was seen in the Aqueous and Petroleum ether extract of Tribu-

lus terrestris. a) P.E-Petroleum ether, (b) AQ-Aqueous, (c) E.A-Ethyl acetate, (d) M-Methanol,

(e) E-Ethanol , (f) CHL-Chloroform, (g) AC-Acetone.

FIGURE 5. MIC of different strains of Petroleum ether extracts of Tribulus terrestris

by broth dilution method

Uma Maheshwari, Rajendran and Vijayalakshmi

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FIGURE 6. MIC of different strains of Aqueous extracts of Tribulus terrestris by broth dilution

method

FIGURE 7. MIC of different strains of Ethyl acetate of Tribulus terrestris by broth dilution

method

gram of STR-VAN-AMX-NEO-CHL-AMP-P-CIP; STR-

VAN-AMX-NEO-AMP-P-GEN-CIP and STR-VAN-CHL-

AMP-P-GEN-AK-CIP.

Totally 10 antibiotics were used to know the resist-

ant pattern, among, 10 antibiogram were recorded. Ten

antibiotics were showing more than 20% resistant to the

isolates. All isolates (100%) were resistant of Ampici-

lin and Penicillin, while Vancomycin showed resistance

(90%), Cipro oxacin (60%), Streptomycin and Amoxyil-

lin (50%), Neomycin (40%), Chlorompenical and Gen-

tamycin (30%) followed by Amikacin (20%). All the

Staphylococcus aureus isolates showed multiple antibi-

otic resistances. Such that one isolate resisted four, six

and seven types of antibiotics, two isolates resisted 3

types of antibiotics, three isolates resisted eight antibi-

otics. The multiple antibiotic resistances (MAR) index

was calculated according to the MAR index formula.

The Maximum MAR index 0.8 was shown by MTUM08,

MTUM06, MTUM07 and minimum MAR index 0.3 was

shown by MBUM01 and MBUM08

Uma Maheshwari, Rajendran and Vijayalakshmi

758 PHARMACOLOGICAL ACTIVITY OF DIFFERENT SOLVENT EXTRACTS OF

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FIGURE 8. MIC of different strains of Methanol of Tribulus terrestris by broth dilution method

FIGURE 9. MIC of different strains of Ethanol of Tribulus terrestris by broth dilution method

Based upon the MDR (Multiple Drug Resistance),

seven strains of Staphylococcus aureus were selected

for further studies. Finally, the strains were tested with

puri ed leaf extract of various solvents using Tribulus

for antimicrobial activity and showed maximum inhi-

bition in Ethyl acetate, moderate inhibition in Ethanol,

minimum inhibition was recorded in Acetone, Chloro-

form, Methanol and no inhibition observed in Aqueous

and Petroleum ether solvents. Minimum inhibitory con-

centrations were performed to determine the minimum

concentration of antibiotics, which is effective on the

wound pathogens were also recorded and compared to

currently used antibiotics, the plant extract of Tribulus

terrestris shows effective results (Fig. 5 to  g. 11).

Post-operative wound infection still remains one of

the most important causes of morbidity and is one of

the most common nosocomial infection (Suljagic et al.,

2010) in surgically treated patients. In the present study,

an attempt has been made to know the predominant

pathogen Staphylococcus aureus is the major respon-

Uma Maheshwari, Rajendran and Vijayalakshmi

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FIGURE 10. MIC of different strains of Methonol of Tribulus terrestris by broth dilution method

FIGURE 11. MIC of different strains of Chloroform of Tribulus terrestris by broth dilution

method

sible for surgical site infections and their antibacterial

susceptibility pattern. The rate of surgical site infection

varies greatly worldwide and from hospital to hospital.

The rate of SSI varies from 2.5% to 41.9% as per differ-

ent studies. The incidence of SSI in the present study is

2.69% even though high, is in agreement with the vari-

ous studies (Reichman and Greenberg, 2009).

Wound infections in ict clinical and societal conse-

quences on the patients, but its bacteriological charac-

teristic varies with different factors. Therefore, effective

treatment and management of wound infections in hos-

pital and community setting will require detailed epide-

miological knowledge of the infecting bacterial patho-

gens and their antibiogram peculiar to the environment.

Uma Maheshwari, Rajendran and Vijayalakshmi

760 PHARMACOLOGICAL ACTIVITY OF DIFFERENT SOLVENT EXTRACTS OF

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Table 2. MIC of different strains of Aqueous extract of Tribulus terrestris by broth dilution method

Test

microorganism

Plant Extract (Aqueous)

Concentration (500mg/ml)

100

(μl)

12.5

(μl)

6.25

(μl)

3.12

(μl)

1.56

(μl)

0.78

(μl)

0.39

(μl)

0.19

(μl)

0.09

(μl)

0.04

(μl)

MTUM02 - + + +++++ ++++

MTUM03 - + + +++++ ++++

MTUM05 - + + +++++ ++++

MTUM06 - + + +++++ ++++

MTUM07 - + + +++++ ++++

MTUM09 - + + +++++ ++++

(-) indicates absence of growth and (+) indicates presence of growth of microorganisms

Table 3. MIC of different strains of Ethyl acetate extract of Tribulus terrestris by broth dilution

method

Test

microorganism

Plant Extract (Ethyl acetate)

Concentration (500mg/ml)

100

(μl)

(μI)

12.5

(μI)

6.25

(μI)

3.12

(μI)

1.56

(μI)

0.78

(μI)

0.39

(μI)

0.19

(μI)

0.09

(μI)

0.04

(μI)

MTUM02 - - - - - - - + + + + +

MTUM03 - - - - - - - + + + + +

MTUM05 - - - - - - + + + + + +

MTUM06 - - - - - - - - - + + +

MTUM07 - - - - - - - - - + + +

MTUM09 - - - - - - - - + + + +

(-) indicates absence of growth and (+) indicates presence of growth of microorganisms

Table 4. MIC of different strains of Methanol extract of Tribulus terrestris by broth dilution

method

Test

microorganism

Plant Extract (Methanol)

Concentration (500mg/ml)

100

(μl)

(μI)

12.5

(μI)

6.25

(μI)

3.12

(μI)

1.56

(μI)

0.78

(μI)

0.39

(μI)

0.19

(μI)

0.09

(μI)

0.04

(μI)

MTUM02 - - - - + + + + + + + +

MTUM03 - - - - - - - + + + + +

MTUM05 - - - - + + + + + + + +

MTUM06 - - - + + + + + + + + +

MTUM07 - - - - - + + + + + + +

MTUM09 - - - - - - + + + + + +

(-) indicates absence of growth and (+) indicates presence of growth Microorganisms

Based on this information, the author (Bularafa Moham-

med Yasidi et al., 2015) examined the prevalence and

antibiogram of bacterial pathogens isolated from wound

infection cases seen at the hospital over the study period.

A total of 392 wound swabs/ and pus of different types

of wound infections from different anatomical sites and

associated clinical conditions were analyzed by standard

bacteriological methods. Of the 392 clinical specimens

analyzed, 301(76.8%) yielded the majority of pathogens

were recovered from septic wound infections. Overall, 7

different bacterial pathogens were identi ed.

The authors state that among different kind of micro-

organisms the major pathogen Staphylococcus aureus

was isolated and used for antimicrobial testing. The

Uma Maheshwari, Rajendran and Vijayalakshmi

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Table 5. MIC of different strains of Ethanol extract of Tribulus terrestris by broth dilution method

Test

microorganism

Plant Extract (Ethanol)

Concentration (500mg/ml)

100

(μl)

(μI)

12.5

(μI)

6.25

(μI)

3.12

(μI)

1.56

(μI)

0.78

(μI)

0.39

(μI)

0.19

(μI)

0.09

(μI)

0.04

(μI)

MTUM02 - - - + + + + + + + + +

MTUM03 - - + + + + + + + + + +

MTUM05 - - - + + + + + + + + +

MTUM06 - - + + + + + + + + + +

MTUM07 - - - + + + + + + + + +

MTUM09 - - + + + + + + + + + +

(-) indicates absence of growth and (+) indicates presence of growth of microorganisms

Table 6. MIC of different strains of Chloroform extract of Tribulus terrestris by broth dilution

method

Test

microorganism

Plant Extract (Chloroform)

Concentration (500mg/ml)

100

(μl)

(μI)

12.5

(μI)

6.25

(μI)

3.12

(μI)

1.56

(μI)

0.78

(μI)

0.39

(μI)

0.19

(μI)

0.09

(μI)

0.04

(μI)

MTUM02 - - + + + + + + + + + +

MTUM03 - - + + + + + + + + + +

MTUM05 - - + + + + + + + + + +

MTUM06 - + + + + + + + + + + +

MTUM07 - - - + + + + + + + + +

MTUM09 - - - - + + + + + + + +

(-) indicates absence of growth and (+) indicates presence of growth of microorganisms

Table 7. MIC of different strains of Acetone extract of Tribulus terrestris by broth dilution

method

Test

Microorganim

Plant Extract (Acetone)

Concentration (500mg/ml)

100

(μl)

(μI)

12.5

(μI)

6.25

(μI)

3.12

(μI)

1.56

(μI)

0.78

(μI)

0.39

(μI)

0.19

(μI)

0.09

(μI)

0.04

(μI)

MTUM02 - - - - - + + + ++++

MTUM03 - - - - + + + + ++++

MTUM05 - - - - - - - - ++++

MTUM06 - - - - - + + + ++++

MTUM07 - - - - + + + + ++++

MTUM09 - - - - - + + + ++++

(-) indicates absence of growth and (+) indicates presence of growth of microorganisms

bacterial pathogens demonstrated high resistance to

Ampicillin (78%), Amoxicillin (66%), and Cotrimoxa-

zole (78%), in contrast to the high sensitivity pattern

observed with O oxacin 83%, Nor oxacin 71%, Cip-

ro oxacin 78%, Erythromycin 72%, Chloramphenicol

62%, Gentamicin 58% and Ceftazidime 60%. But in

this present study demonstrates that all Staphylococcus

aureus isolated from pus samples of infectious patients.

The high resistance was observed in Cipro oxacin (60%),

Amoxicillin (50%) in contrast to high sensitivity pat-

tern observed with Chloramphenicol (30%), Gentamicin

(30%), Nor oxacin (40%).

Jain et al. (2014) states that in their study all Staphy-

lococci were susceptible to Vancomycin and Teicoplanin.

Uma Maheshwari, Rajendran and Vijayalakshmi

762 PHARMACOLOGICAL ACTIVITY OF DIFFERENT SOLVENT EXTRACTS OF

TRIBULUS TERRESTRIS

BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

In contrast, the Vancomycin remains the  rst choice of

treatment for MRSA and to preserve its value, its use

should be limited to those cases where there are clear

indications. But in this present study dissimilar results

were obtained by predominant pathogen Staphylococ-

cus aureus indicates resistant to Vancomycin, it proofs

that the emerging of drug resistant ability of Staph to be

currently initiated.

The result of this present study indicated that bacte-

rial isolates demonstrated high sensitivity to Chloram-

phenicol being about 70% sensitive, whereas resistant

to -lactam antibiotic namely Penicillin (100%) was

very high. These results are contrary to that obtained for

anaerobes isolated from oro-facial infections in earlier

study which reported good activities of the later agents

against the anaerobes (Nicholes, 2004).

Recurrently, the medicinal plants are the oldest form

of healthcare known to mankind. From the ancient time

people are using different herbs or plants as the rem-

edy forvarious diseases. But now a day’s people have

become dependent on synthetic medicines which have

many side effects. So to reduce the side effects we can

use medicinal plants for the treatment of common dis-

eases rather than using drugs.

Phan et al. (2001)  nd out the secondary metabolites

are synthesized by the plant as part of the defence sys-

tem of the plant. The plant contains chebulinc acid, tan-

nic acid, gallic acid, resin, anthroquinone and sennoside.

It also contains glycosides, sugar, terpenoids, steroids,

phosphoric acid and these compounds were proven to

exhibit antibacterial, antifungal, antiviral and anticar-

cinogenic (Neamsuvan et al., 2012). Hence, the present

study to logically select a leaf part of bio-medically

active Tribulus plant to exhibit a broad antimicrobial

activity against wound pathogen Staphylococcus aureus.

Ali et al., (2001) con rmed that their study the Tribulus

terrestris had no detectable antibacterial activity against

any of the infectious disease causing reference bacte-

ria. For this reason, in this present study choose a plant

material of Tribulus to check the vital activity against

infectious disease that proved the Indian leaf of T. ter-

restris were active against clinically dominant pathogen

S.aureus isolated from post-operative wound infection.

Recurrently, the plant extract is highly sensitive when

compared to the standard antibiotic. These data indi-

cate that Gram-positive bacteria are the most sensitive

strains for the different extracts, which exhibited their

main antibacterial activity on Gram-negative bacte-

ria (Bakri and Douglas 2005). Our results are in good

agreement with previous works (Al Maofari et al., 2013)

showing a weaker activity of essential oil of Anis and

Salvia tomentosa and also Nair and Chanda, a (2008)

also reported Gram positive were more sensitive. Etha-

nol extract showed more effect than aqueous extract in

inhibiting the growth of the bacterial strains (Nair and

Chanda, b 2007; Firas et al., 2008). The growth of the

Gram positive strains was found more inhibition than

Gram negative. In this present study, similar results was

observed the antimicrobial activity of 7 different solvent

plant extract was tested against S.aureus. The maximum

antibacterial activity was seen in Ethyl acetate extract

(100μl) of Tribulus terrestris. Intermediate antibacterial

activity was seen in Ethanol extract (100μl) of Tribulus

terrestris (Fig. 3).

CONCLUSION

The present study revealed that the post operative wound

infection is one of the most common chronic infectious

diseases of human is also a serious problem among wound

of post-operative patients caused by predominant patho-

gen Staphylococcus aureus. Therefore, effective treatment

and management of wound infections in hospital and

community setting will require detailed knowledge of the

infecting bacterial pathogens and their antibiogram pecu-

liar to the environment for drug selection. Resistant bac-

teria which are human pathogens may cause the disease,

they may still be dangerous because they can transfer

their antibiotic resistance genes to other organism. Anti-

microbial resistances of bacterial pathogens are a major

problem for the treatment of animal and human patients

with bacterial diseases. There is an alarming increase

of infections caused by antibiotic resistant bacteria to

urgently need of antimicrobial compound to treat a path-

ogenic organism. Recurrently the plants have been one of

the important sources of medicine since the beginning of

human civilization. There is a growing demand for plant

based medicines to activity against several infectious

pathogens. Hence the current study concluded that the

different solvent extract of Tribulus plant has different

mode of action to eradicate the wound pathogen around

the surgical site. The plant extracts can also be exploited

in designing the wound care products.

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