Wageningen 

Academic 
Beneficial Microbes, September 2011; 2(3): 235-243 P u b l i s h e r s

Antibacterial activity of selected medicinal plants against multiple antibiotic resistant

uropathogens: a study from Kolli Hills, Tamil Nadu, India
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A.S. Narayanan1, S.S.S. Raja2, K. Ponmurugan1, S.C. Kandekar3, K. Natarajaseenivasan4, A. Maripandi5 and
Q.A. Mandeel6

1K.S. Rangasamy College of Arts and Science, PG and Research Department of Microbiology, KSR Kalvi Nagar, Thokkavadi

Post, Tiruchengode 637 215, Tamil Nadu, India; 2Bharathidasan University College, Department of Microbiology, Perambalur,
Tiruchirapalli, Tamil Nadu, India; 3KSR Institute of Dental Science & Research, KSR Kalvi Nagar, Thokkavadi Post,
Tiruchengode 637 215, Tamil Nadu, India; 4Bharathidasan University, Department of Microbiology, Tiruchirapalli 620 024,
Tamil Nadu, India; 5Government Arts College, Dept of Microbiology, Dharmapuri 636 705, Tamil Nadu, India; 6University
of Bahrain, Department of Biology, P.O. Box 32038, Sakhir Campus, Bahrain; san_kamal_4@yahoo.com

Received: 4 August 2010 / Accepted: 26 August 2011

© 2011 Wageningen Academic Publishers

Abstract

The increasing incidence of antibiotic resistance among bacterial pathogens necessitates medicinal plants as an
alternate therapy in restricting the resistant infectious organisms. In this primitive study, the antibiotic resistance
of organisms isolated from urinary tract infected patients was evaluated using the National Committee for Clinical
Laboratory Standards (NCCLS) method and Multiple Antibiotic Resistance (MAR) index values, and the MAR values
was also calculated for plant extracts. The 10 common medicinal plants collected from Kolli hills, Namakkal, south
India were extracted using the chloroform, methanol, acetone, ethanol and saponification procedure. The efficacy
of the extracts on the uropathogens was tested by agar disc diffusion method in order to analyse the inhibitory
activity of plant extract on the organisms. Azadiracta indica A. Juss., Tinospora cordifolia (Wild.) and Euphorbia
hirta Linn. exhibited high inhibitory activity against most of the 11 tested organisms followed by Cassia javanica
Linn. and Phyllanthus niruri Linn. The maximum zone size of 46.3 mm was exhibited by methanol extract of
P. niruri Linn. against Pseudomonas aeruginosa. Asparagus racemosus Willd. and Eupatorium triplinerve Vahl had
the least activity against resistant pathogens. Saponified lipids of most of the plants exhibited maximum antibacterial
activity. Among the tested organisms, P. aeruginosa and Staphylococcus epidermidis were the most susceptible and
Serratia marcescens, Enterobacter cloaceae, Citrobacter koseri, and Citrobacter freundii were the least inhibited by
most of the extracts of medicinal plants. It is concluded that revised antibiotic policies and more importantly the
development of herbal medicine as an alternative may be incorporated in urological practice.

Keywords: antibacterial activity, multiple antibiotic resistance, uropathogens, Azadiracta indica, Tinospora cordifolia,
Euphorbia hirta

1. Introduction has become more complicated because of the appearance of

Urinary tract infections (UTI) are the most common form
of bacterial infection affecting people throughout their
life (Barnett and Stephen, 1997; Foxman, 2002). It is one
of the common reasons for adults to seek medical help
and is one of the most frequently occurring nosocomial
infections (Gastmeier et al., 1998). The treatment of UTI
pathogens with increasing resistance to antimicrobial agents
(Murakami et al., 1995). Such antibiotic resistant organisms
are difficult to eliminate during the course of an infection
and are lethal to the human population. In addition,
antibiotics in these cases are sometimes associated with
adverse effects on the host, which include hypersensitivity,
depletion of beneficial gut and mucosal microorganism,

ISSN 1876-2833 print, ISSN 1876-2891 online, DOI 10.3920/BM2010.0033 235

 A.S. Narayanan et al.
immunosuppressant and allergic reactions (Idose et al.,
1968). Therefore, there is a need to develop alternative
antimicrobial drugs for the treatment of infectious diseases
in general and of UTI in particular.
During the last decade, the use of traditional medicine has
expanded globally and is gaining popularity as an alternative
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medicine. It has continued to be used not only for primary
health care of the poor in developing countries, but also
in countries where conventional medicine is predominant
in the national health care system (Lanfranco, 1999).
According to the World Health Organization (WHO), herbal
medicines serve the health needs of about 80% of the world’s
population, especially for millions of people in the vast
rural areas of developing countries (WHO, 2001). Hence,
pharmacognostic investigations of plants are carried out to
find novel drugs or templates for the development of new
therapeutic agents. In spite of intensive investigation, only
5-10% of more than 250,000 species of higher plants have
been chemically investigated (Nahrstedt, 1996). There is an
increasing need for the documentation and development
of alternative anti-pathogenic substances. Accordingly, the
antimicrobial properties of certain Indian medicinal plants
were reported based on folklore information (Ashmad et al.,
1998; Dayal and Purohit, 1971; Hook and Thomas, 1995;
Mehmood et al., 1999; Reddy, 1995; Suresh et al., 1995)
and a few attempts were made to study inhibitory activity
against certain pathogenic bacteria and fungi (Taylor et al.,
1995). Ayyanar and Ignacimuthu (2005), Duraipandiyan
et al. (2006), Natarajan et al. (1999), Rajan et al. (2002)
and Sandhya et al. (2006) reported the practice of green
pharmacy by the population in Tamil Nadu state of India.
Considering the frequent emergence of antibiotic resistant
strains and the immense potential of traditional medicines
of plant origin, this work has been designed to study the
efficacy of 10 medicinal plants of the Kolli hills of Namakkal,
south India, against 11 multiple antibiotic resistant bacterial
strains isolated from infected persons. The medicinal
plants were selected based on folkloric medicine used by
traditional healers dwelling at the foot of the Kolli hills. The
significance of the present study lies in the fact that this is
the first of its kind in evaluating the efficiency of indigenous
plants species against multiple antibiotic resistant bacterial
strains causing nosocomial UTI with significant Multiple
Antibiotic Resistant (MAR) values.
2. Materials and methods
Study area
The Kolli hills, a part of Eastern ghat situated in the district
of Namakkal, Tamil Nadu, south India lies between 11°10’
to 11°30’N and 78°15’ to 78°30’. It has an area of 482 km2
and stretches 29 km from north to south and 19 km from
east to west. The highest point in the Kolli hills is 1,380
236
meter above sea level, but the general level of the upper
surface of the hill is not more than 1000 m (average rainfall
942 mm and temperature 27 °C during study period). The
bioresource, especially the flora, is largely unexplored for
its identity and biological functions.
Plant collection
The medicinal plants included in the Table 1 were collected
fresh from the Kolli hills during the period of June 2006
– March 2007 and maintained in the KSR medicinal
garden in college premises. The plants were identified and
authenticated by Prof. P. Ponmurugan (PG Department of
Biotechnology, KSR College of Technology, Tiruchengode,
Namakkal, India) and preserved in our college herbarium.
The identifications were based on the descriptions by
Chatterjee et al. (1997). The leaves, stems and roots of
medicinal plants were washed in running tap water to clean
the adhering particles, sterilised with 0.1% mercuric chloride
for 1 min and washed again with distilled water. Then the
plants were subjected to the antibacterial activity tests.
Bacterial isolates and their multiple antibiotic resistance
index
Three Gram positive and eight Gram negative organisms
included in this study are indicated in Table 2. The
pathogenic organisms were isolated from urine specimen
of persons suffering from UTI and identified by standard
methods (Koneman et al., 1997). The cultures were
maintained in the department laboratory. The antibiotic
susceptibility patterns of the test organisms were performed
using standard procedure (NCCLS, 2003) and their
MAR index was calculated by the ratio of the number of
ineffective antibiotics to the total number of antibiotics
tested (Krumperman, 1983) against different numbers of
isolates. The MAR index value was also calculated for the
herbal extracts. The various concentrations of antibiotics
as per the practical applications for in vitro studies have
been employed (Lee et al., 2009; Samie et al., 2005). The
antibiotics (Himedia, Mumbai, India) used in the study
were: ampicillin (10 µg), amoxicillin (30 µg), amikacin
(30 µg), cephalothin (30 µg), chloramphenicol (30 µg),
ciprofloxacin (5 µg), ceftriaxone (30 µg), clindamycin (2 µg),
co-trimoxazole (1.25 µg), erythromycin (15 µg), gentamycin
(10 µg), kanamycin (30 µg), methicillin (5 µg), novobiocin
(30 µg), nalidixic acid (30 µg), norfloxacin (10 µg), ofloxacin
(5 µg), penicillin G (10 units), rifampicin (5 µg), streptomycin
(10 µg), tetracyclin (30 µg) and vancomycin (30 µg). The
reference strains used for the study were Escherichia
coli ATCC 25922, Staphylococcus aureus ATCC 29213,
Pseudomonas aeruginosa ATCC 27853, Enterococcus faecalis
ATCC 29212, Staphylococcus epidermis ATCC 14990,
Klebsiella pneumoniae ATCC 12658, Serratia marcescens
ATCC 274, Proteus mirabilis ATCC 35491, Citrobacter
freundii ATCC 8090 and Enterobacter cloacae ATCC 10699.
Beneficial Microbes 2(3)
  Antibacterial activity of medicinal plants against uropathogens

Table 1. Antibacterial activity of the crude medicinal plant extracts (40 µg/disc) against antibiotic resistant uropathogens.

Plant name Parts Solvent3 Bacterial species1,2 (mean inhibition zone in mm)
(local name) used
Se Ef Sa Pa Kp Ecl Pm Sm Cf Ck Ec

Phyllanthus fraternus leaves e 25.1 - 20.4 32.4 - - - - - - -

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Webster m 25.4 - 18.1 46.3 - - - - - - -

(Keezhanelli) a 20.2 - - - - - - - - - -
c - - - - - - - - - - -
s 18.1 20.1 15.2 19 12.1 15.5 - 15.6 10.8 8.1 9.3
Asparagus racemosus roots e - - - 40.1 - - - - - - -
Willd. m - - - - - - - - - - -
(Thaneervittan kizhangu) a 18.2 - - - - - - - - - -
c - - - - - - - - - -
s 20.1 22 14.4 - - 13.2 20.1 15.7 - 8.2 -
Eupatorium triplinerve Vahl leaves e 25.1 - - - - - - - - - -
(Ayapana) m 20.3 15.6 - - - - - - - - -
a 16.2 12.1 - 20.7 - - - - - -
c - - - - - - - - - - -
s 18.1 18.1 - - - - - 14.3 - - -
Leucas aspera Spreng. leaves e - - - 35.1 - - - - - - -
(Thumbai) m - - - 40.3 - - - - - - -
a - - - 25 - - - - - - -
c - - - 30 - - - - - - -
s 17.1 20.1 15.1 18.1 - - 20.2 17.4 - 9.6 8.1
Glycosmis pentaphyllia Ritz leaves e - - 15.1 - - - - - - - -
(Kuttivila) m - - 13.2 - - - - - - - -
a - - - - - - - - - - -
c - - - - - - - - - - -
s 22 18.1 - 13.2 - - 18.1 17.3 9.3 - 7
Azadirachta indica A. Juss. leaves e 17 7 16 5 20 7 8 - - - -
(Vembu) m 7 7 7 7 5 10 12 - - 9 -
a 8 7 14 - 8 9 7 - - - 10
c 12 8 7 9 18 12 8 - - - -
s 28 22 20 24 22 32 23 18 - 10 -
Tinospora cordifolia Wild. stem e 7 8 7 9 10 7 - - - 7 -
(Shindilakodi) m 7 5 8 7 7 8 8 - - - -
a 7 10 12 13 11 13 11 - 9 - -
c 11 14 15 8 11 9 12 - - 9 -
s 32 19 24 19 22 27 22 17 9 10 11
Euphorbia hirta Linn. leaves e 11 10 14 15 13 17 14 - - - -
(Amman pacharisi) m 14 - 18 13 12 17 16 - - 11 -
a 12 13 18 14 12 14 15 - - - 9
c - - - - - - - - 10 - -
s 24 20 23 14 16 17 19 18 10 11 8
Phyllanthus niruri Linn. leaves e 18 - 14.2 18.1 - - - - - - -
(Keezhanelli) m 19.1 - 15.3 22.2 - 15.1 - - - - -
a 15 - 19.1 - - - - - - - -
c - - - - - - - - - - -
s 16.1 20.4 17 16 21.9 24.1 20.3 13.2 12.2 8 -
Cassia javanica Linn. seeds e - 15 - - 21.1 - - - - - -
(Konrai) m - - - 23.1 16.3 - - - - - -
a - - 9.2 19.5 - - - - - - -
c - - 11 14.4 9.4 - - - - - -
s 11.1 - 14.5 18.3 13 - - 21.1 - 7.2 9.3

1 Bacterial species: Staphylococcus epidermidis (Se), Enterococcus faecalis (Ef), Staphylococcus aureus (Sa), Pseudomonas aeruginosa (Pa), Klebsiella

pneumoniae (Kp), Escherichia coli (Ecl), Proteus mirabilis (Pm), Serratia marcescens (Sm), Citrobacter freundii (Cf), Citrobacter koseri (Ck), Enterobacter
cloacae (Ec).
2 Zone of inhibition includes the diameter of the disc (6 mm); - no zone formation.
3 Solvents used: E = ethanol; M = methanol; A = acetone; C = chloroform; S = saponified lipid.

Beneficial Microbes 2(3) 237

 A.S. Narayanan et al.

Table 2. Multiple antibiotic resistance index values of antibiotic resistant organisms and number of isolates used in this study.

Organism Number of MAR value1 Number of ineffective Name of ineffective antibiotics2

isolates antibiotics

Staphylococcus epidermidis 9 0.2 5 S, T, Nv, C, Ch

Enterococcus faecalis 9 0.41 9 Na, E, Va, T, Of, Nx, Ci, C, Ak
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Staphylococcus aureus 9 0.64 14 S, T, Nv, C, Ch, M, A, Ac, Na, Nx, P, Ci, G, Ch

Pseudomonas aeruginosa
Klebsiella pneumoniae
Escherichia coli
Proteus mirabilis
Serratia marcescens
Citrobacter freundii
Citrobacter koseri
Enterobacter cloacae
9 0.32
8 0.36
10 0.5
2 0.32
1 0.24
1 0.64
1 0.73
1 0.91
7 A, Ac, Ci, G, Of, P, C
8 A, Ac, Ci, G, Of, P, C, Cd
11 A, Ac, Ci, G, Of, P, C, Cd, Ak, Ch, T
7 A, Ac, Ci, G, Of, P, C
6 A, Ac, Ci, G, Of, P,
14 S, T, C, Ch, M, A, Ac, Cf, Ak, Of, P, Ci, G, Ch
16 S, T, C, Ch, M, A, Ac, Cf, Ak, Of, P, Ci, G, Ch, R, Va
20 S, T, Nv, C, Ch, M, A, Ac, Na, Nx, P, Ci, G, Ch, K,
Cf, Co, Va, E, G

1 A multiple antibiotic resistance (MAR) value near to 1 indicates the antibiotics are ineffective.
2 Antibiotic abbreviations: ampicillin (A), amoxicillin (Ac), amikacin (Ak), cephalothin (Ch), chloramphenicol (C), ciprofloxacin (Cf), ceftriaxone (Ci),
clindamycin (Cd), co-trimoxazole (Co), erythromycin (E), gentamycin (G), kanamycin (K), methicillin (M), novobiocin (Nv), nalidixic acid (Na), norfloxacin
(Nx), ofloxacin (Of), penicillin G (P), rifampicin (R), streptomycin (S-10 µg), tetracycline (T), vancomycin (Va).

Preparation of extract and discs
Fresh matured leaves, stems, seeds and roots of plant
specimens were dried in shade and powdered by a
mechanical mortar separately. Powdered plant materials
(10 g) were used for extraction of the bioactive compounds
in Soxhlet extractor with organic solvents chloroform,
methanol, acetone or ethanol. After concentrating the
extract at 50 °C for 6 h it was stored at 4 °C until further
use. For the extraction of saponified lipid content of
medicinal plants, a modified saponification process as
applied to plants was employed (Padmini et al., 1986).
For preparing the plant extract disc, 1 ml of concentrated
extract was incorporated into the pre-weighed watch glass
and evaporated by heat at 50-60 °C for 1 hour and weighed
again. Thus, the weight of the bioactive component in
1 ml was calculated and hence different concentrations
were loaded onto sterile discs (Himedia, Mumbai, India)
to reach 40 μg. For the present study, herbal extract discs
were prepared at concentrations of 10, 20, 30 (data not
shown) and 40 μg.
Determination of antimicrobial activity of solvent
extracted medicinal plants
Two sets of Mueller Hinton agar (HiMedia) plates were
prepared and seeded with 16 h broth cultures of different
pathogenic organisms. Before making the lawn culture
with the inoculum, the turbidity was adjusted to 0.5% of
McFarland solution (1-2×107 cfu/ml). Seeded inoculum
was treated with different concentrations of herbal extract
discs. The plates were incubated at 37 °C for 24 h. The
experiment was repeated twice and the average of the zone
of inhibition was recorded.
3. Results and discussion
The MAR index values of the test organisms are reported
in Table 2 and the MAR index value for herbal extracts are
reported in Table 3. The MAR value is a ratio of the number
of effective antibiotics to that of ineffective antibiotics tested
against the different number of isolates. The antibacterial
activities and minimum inhibitory concentration values of
different solvent extract of 10 common medicinal plants of
Kolli hills are given in Tables 1 and 4. Azadirachta indica,
Tinospora cordifolia and Euphorbia hirta exhibited a high
degree of inhibitory activity against most of the 11 tested
organisms followed by Cassia javanica and Phyllanthus
niruri. Saponified lipids of these plants exhibited
maximum activity. Among the pathogens, P. aeruginosa
and S. epidermidis were the most susceptible followed by S.
aureus and Enterococcus faecalis, E. cloaceae, S. marcescens,
Citrobacter koseri and C. freundii were the least inhibited
by most of the extracts of medicinal plants. Due to the
over-usage of antibiotics (Sydney et al., 1980), mutation and
environmental stress the antibiotic resistant organisms have
become a major challenge in hospital acquired infections.
This often causes difficulties for the treatment of UTI
patients. Because of this drug resistance, the search for
new antibiotics continues unabated. In this connection,
plants continue to be a rich source of therapeutic drugs.
The active ingredients of many drugs are found in plants, or

238 Beneficial Microbes 2(3)

  Antibacterial activity of medicinal plants against uropathogens

Table 3. Multiple antibiotic resistance index values of herbal CDCP (2002) identified a strain of methicillin resistant S.
plant extracts on antibiotic resistant organisms used in this aureus (MRSA) from United States which was resistant
study. to arbekacin and vancomycin. In our study the organism
has shown significant MAR value of 0.64 but has been
Name of the organism No. of No. of MAR susceptible to the saponified lipids of T. cordifolia (24 mm)
extracts extracts not value1 and E. hirta (23 mm).
used inhibiting
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the growth P. aeruginosa susceptibility to antimicrobials, especially

piperacillin, ceftazidime and imipenim/cilastatin, has shown
Staphylococcus epidermidis 50 19 0.38 a decreasing trend over the past 20 years (Shigemura et
Enterococcus faecalis 50 28 0.56 al., 2005). The often discussed multi-drug-resistant
Staphylococcus aureus 50 22 0.44 P. aeruginosa is generally considered to be resistant to
Pseudomonas aeruginosa 50 19 0.38 ciprofloxacin, impenem/cilastatin and amikacin (Arakawa,
Klebsiella pneumoniae 50 30 0.60 2002). Accordingly, the organism in the present study
Escherichia coli 50 32 0.64 exhibited a MAR value of 0.32. In our study the ethanol
Proteus mirabilis 50 33 0.66 and methanol extracts of P. niruri (18.1 and 22.2 mm
Serratia marcescens 50 40 0.80 respectively) and Leucas aspera (35.1 and 40.3 mm
Citrobacter freundii 50 43 0.86 respectively), ethanol extracts of A. racemosus (40.1 mm)
Citrobacter koseri 50 38 0.76 and saponified lipids of all the plants inhibited P. aeruginosa
Enterobacter cloacae 50 42 0.84 except A. racemosus and E. triplinerve. Organisms exhibited
susceptibility to the herbal extract in general and methanol
1 A multipleantibiotic resistance (MAR) value near to 1 indicates that extracts in particular. Methanol is more efficient than
the extracts are ineffective. acetone in extracting phytochemicals from plant materials
(Cowan, 1999; Eloff, 1998). Plants contained microbial
inhibitors, i.e. flavonoids soluble in aqueous methanol
(Otshudi et al., 1999). Methanol extracted more inhibitors
are produced as secondary metabolites and are increasingly from plant (Ojala et al., 2000). This could be the reason for
popular among village communities because there is real higher susceptibility of the organism in methanol extract.
improvement of disease conditions after herbal treatment
and harmful side effects and the high cost of other forms A study pertaining to the incidence of uropathogens has
of treatment are very much reduced. found that Klebsiella on average accounts for 10% of
uropathogens in Europe (Wagenlehner and Naber, 2004).
Methicillin resistant S. epidermidis (MRSE) in urinary Gram negative bacteria, which include Klebsiella spp.,
isolates is responsible for complicated UTI (Sakumoto et were resistant to broad spectrum antibiotics prescribed
al., 1996). In our study, S. epidermidis exhibited a MAR for UTI; the percentage of resistance ranged from 2.8% for
value of 0.2 but P. niruri, Eupatorium triplinerve, A. indica, imipenem to 15% for ceftazidime (Bouza et al., 2001). In
T. cordifolia, E. hirta and P. niruri exhibited inhibitory our study K. pneumoniae exhibited a MAR value of 0.36.
activity over the organism. The maximum inhibitory zone But these isolates were sensitive to ethanolic extracts of A.
of 32 and 28 mm was exhibited by saponified lipid of T. indica (20 mm) and C. javanica (21.1 mm) and saponified
cordifolia and A. indica respectively. lipids of A. indica (22 mm), T. cordifolia (22 mm) and P.
niruri (21.9 mm).
E. faecalis susceptibilities to erythromycin and minocycline
have undergone significant changes; the former decreased The isolation rate of ciprofloxacin resistant E. coli ranged
significantly in the last 5 years (P<0.05) and the latter from 18.1 to 18.9% in 1997-2004 (Gales et al., 2000). In
increased significantly in the last 10 years (P<0.05) accordance with this study, the isolate E. coli in our finding
(Shigemura et al., 2005). Nevertheless, one cannot overlook has exhibited a significant MAR value of 0.5 indicating
the possible emergence of resistant strains because of the its high degree of acquired resistance. The fact that the
frequent use of these antimicrobial agents to manage the organism was inhibited by saponified lipids of A. indica (32
frequently isolated E. faecalis. However, in our study, the mm), T. cordifolia (27 mm) and P. niruri (24.1 mm) suggests
organism exhibited a MAR value of 0.41 and susceptibility its inherent susceptibility to the tested plant extracts.
was observed against the saponified lipid of all the plants
tested except C. javanica, the maximum being that of A. In a study, E. coli was found to be the most insensitive
indica and Asparagus racemosus (22 mm). strain of all the tested bacteria to herbal extracts (Tadeg et
al., 2005). In fact, most Gram negative bacteria reported
S. aureus susceptibility to vancomycin and arbekacin to have developed multidrug resistance to many of the
has remained nearly 100%. However, a report from the antibiotics currently available on the market, of which E.

Beneficial Microbes 2(3) 239

 A.S. Narayanan et al.

Table 4. Minimum inhibitory concentration (mean±SD) of crude medicinal plant extracts against antibiotic resistant uropathogens.

Medicinal plant Solvent2 Minimum inhibitory concentration (µg/ml)1

extract
Se Ef Sa Pa Kp Ecl Pm Sm Cf Ck Ec

Phyllanthus e 20±1.1 - 34±1.3 13.5±1.1 - - - - - - -

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fraternus m 22±1.1 - 36±1.1 6±1.1 - - - - - - -

a 28±1.6 - - - - - - - - - -
c - - - - - - - - - - -
s 32±1.1 29±1.1 52±1.4 31±1.3 54±1.1 56±1.1 - 51±1.1 80±1.3 90±1.6 72±1.1
Asparagus e - - - 40.1±1.6 - - - - - - -
racemosus m - - - - - - - - - - -
a 33±1.1 - - - - - - - - - -
c - - - - - - - - - -
s 29±1.7 35±1.1 48±1.1 - - 75±1.1 30±1.3 40±1.3 - 80±1.3 -
Eupatorium e 27±1.3 - - - - - - - - - -
triplinerve m 33±1.3 40±1.6 - - - - - - - - -
a 39±1.4 59±1.1 - 28±1.1 - - - - - -
c - - - - - - - - - - -
s 35±1.4 31±1.6 - - - - - 50±1.9 - - -
Leucas aspera e - - - 15±1.1 - - - - - - -
m - - - 14±1.1 - - - - - - -
a - - - 18±1.1 - - - - - - -
c - - - 12±1.1 - - - - - - -
s 48±1.1 30±1.6 54±1.1 37±1 - - 46±1.1 55±1.1 - 60±1.1 70±1.6
Glycosmis e - - 60±1.6 - - - - - - - -
pentaphyllia m - - 68±1.1 - - - - - - - -
a - - - - - - - - - - -
c - - - - - - - - - - -
s 26±1.6 57±1.6 - 60±1.1 - - 48±1.1 46±1.1 88±1.1 - 90±1.1
Azadirachta e 52±1.1 94±1.6 40±1.6 95±1.1 38±1.1 92±1.6 94±1.1 - - - -
indica A m 92±1.1 95±1.1 94±1.1 96±1.5 95±1.1 80±1.3 82±1.1 - - 82±1 -
a 90±1.1 96±1.1 48±1.1 - 88±1.1 91±1.1 96±1.6 - - - 80±1.6
c 75±1.1 90±1.1 94±1.9 90±1.6 38±1.1 58±1.1 94±1.1 - - - -
s 22±1.6 38±1.1 30±1.1 36±1 41±1.6 14±1.3 28±1.1 45±1.1 - 78±1.6 -
Tinospora e 97±1.6 94±1.3 97±1 91±1.6 80±1.1 94±1.1 - - - 91±1.1 -
cordifolia m 97±1.1 95±1.1 96±1.6 95±1.1 86±1.1 90±1.1 95±1.1 - - - -
a 97±1.1 80±1.6 78±1.6 69±1.3 78±1.1 74±1.1 82±1.1 - 80±1.1 - -
c 78±1.1 74±1.1 50±1.1 88±1.1 77±1.1 90±1.1 77±0.8 - - 76±1.1 -
s 8±1.3 52±1.6 44±1.1 49±1 45±1.3 32±1.7 48±1 42±1.1 80±1.9 84±1.3 80±1.1
Euphorbia hirta e 54±1.3 58±1.1 48±1.1 40±1.1 44±1.1 36±1.3 44±1.1 - - - -
m 60±1.1 - 42±1.5 32±1.1 52±1.6 34±1.4 42±1.3 - - 11±1.6 -
a 52±1.1 53±1.6 38±1.1 49±1.6 50±1.6 32±1.9 52±1.5 - - - 9±1.1
c - - - - - - - - 10±1.6 - -
s 30±1.1 6.25±1.3 28±1.1 43±1.6 38±1.6 36±1.1 35±1.7 32±1.1 58±1.3 56±1.5 60±1.4
Phyllanthus niruri e 32±1.1 - 42±1.3 40±1.7 - - - - - - -
m 34±1.1 - 44±1.9 42±1.5 - 58±1.8 - - - - -
a 42±1.3 - 51±1.9 - - - - - - - -
c - - - - - - - - - - -
s 42±1.3 38±1.6 45±1.6 51±1.9 42±1.5 38±1.3 42±1.1 56±1.1 58±1.6 58±1.6 -
Cassia javanica e - 52±1.6 - - 42±1.5 - - - - - -
m - - - 28±1.1 41±1.6 - - - - - -
a - - 62±1.8 35±1.1 - - - - - - -
c - - 58±1.1 44±1.1 60±1.7 - - - - - -
s 58±1.1 - 54±1.6 38±1.1 55±1.1 - - 31±1.6 - 92±1.1 83±1.9

1 Bacterial species: Staphylococcus epidermidis (Se), Enterococcus faecalis (Ef), Staphylococcus aureus (Sa), Pseudomonas aeruginosa (Pa), Klebsiella

pneumoniae (Kp), Escherichia coli (Ecl), Proteus mirabilis (Pm), Serratia marcescens (Sm), Citrobacter freundii (Cf), Citrobacter koseri (Ck), Enterobacter
cloacae (Ec); - not done.
2 Solvents used: E = ethanol; M = methanol; A = acetone; C = chloroform; S = saponified lipid.

240 Beneficial Microbes 2(3)

  Antibacterial activity of medicinal plants against uropathogens
coli is prominent (Alonso et al., 2000). However, the plants
in our study are of special interest for further investigation
since they showed exceptionally strong activity against E.
coli, especially A. indica.
P. mirabilis accounts for 7% of uropathogens in Europe
and its resistance to cefuroxime ranged between 9-17%
http://www.wageningenacademic.com/doi/pdf/10.3920/BM2010.0033 - Saturday, October 07, 2017 8:28:41 AM - Göteborgs Universitet IP Address:130.241.16.16
(Wagenlehner and Naber, 2004). In our study, Proteus
exhibited a MAR value of 0.32. Again, these organisms
were susceptible to saponified lipid extracts of A. indica
(23 mm), T. cordifolia (22 mm), E. hirta (19 mm) and P.
niruri (20.3 mm).
In a study by Gootz et al. (1984), wild clinical strains of C.
freundii were found to be resistant to cephalothin, cefoxitin,
cefoperazone, cefotaxime and cefamandole. In our study
the MAR value of this organism was higher than 0.64.
But the organism was inhibited by chloroform extract
and saponified lipids of E. hirta (10 mm each), saponified
lipids of P. niruri extract (12.2 mm) and acetone extract
and saponified lipids of T. cordifolia (9 mm).
The drug resistance of C. koseri was found to be higher
than other Citrobacter species in their analysis of various
clinical samples including urine in hospitals of North
India (Mohanty et al., 2007). In the present investigation
C. koseri had a MAR index value of 0.73 and was inhibited
by methanol and saponified lipids of E. hirta (11 mm).
Generally, antibiotic resistance is defined, if bacteria can
still grow under achievable therapeutic concentrations of
antibiotic substances at the site of infection. Resistance
is related to the amount of application of an antibiotic
substance and is divided into primary or inherent resistance
against an antibacterial substance and secondary or
acquired resistance, if resistance emerges in intrinsically
susceptible bacteria. Bacteria do exhibit an enormous
repertoire of different resistance mechanisms. Unspecific
mechanisms such as reduced permeability or efflux alter
the tolerance to antibiotics substances less than the specific
mechanism such as inactivation of antibiotics or alteration
of target structures. By and large, resistance is a cumulative
effect of all these three factors as in Pseudomonas spp.
where the combination of gentamicin modifying enzymes,
powerful efflux mechanism and decreased permeability is
more common, or E. coli where alteration in target gyrase
enzymes and production of beta-lactamase is common
(Wagenlehner and Naber, 2004).
However, according to earlier reports (Cowan, 1999;
Thomson, 1978) the reasons for resistance could be
predicted, the high degree of susceptibility to the plant
extract in our study cannot be stated without experimental
evidence. One reason seems obvious. Basically the Gram
negative cell wall due to the presence of lipopolysaccharides
(LPS) in outer membrane is impermeable to antibiotic and
Beneficial Microbes 2(3)
hence, it is resistant to multiple antibiotics. Diffusion of
plant extracts has not been altered by the impermeable
LPS of Gram negative organisms in our study. Hence,
permeability can be considered as one reason for the
efficiency of plant extracts in our study. The bioactive
principle in plant and their mode of action needs further
evaluation. Furthermore, the plant part, solvent and the
methods used to extract the bioactive components played
an important role in its potential antibacterial activity
(Cowan, 1999; Thomson, 1978). It may be a reason for
variation of the antibacterial activity of medicinal plants
used in this study. The bactericidal properties of fatty acids
are well known (Bayliss, 1936; Kodicek, 1949; Nieman,
1954). Similarly, in the present investigation effective results
were obtained in the saponified lipids against all the tested
pathogenic organisms. However, the mode of action of
fatty acids cannot be wholly explained in physiochemical
terms. Drug action may involve a change in permeability
of the cell wall or interference with cellular metabolism
(Kabara et al., 1972).
To conclude, antibiotic resistance is an increasing problem
in urological practice. Nosocomial uropathogens in
particular may exhibit resistance to multiple antibiotics
and pose problems for empirical therapy. In future, the
rate of antibiotic resistance may well continue to increase.
Strategies to slow down this trend, such as revised antibiotic
policies and more importantly the development of herbal
medicine as an alternate may be incorporated in urological
practice.
Acknowledgements
The authors are grateful to the Management and Principal
of K.S. Rangasamy College of Arts and Science for providing
the necessary facilities.
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