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Seema Rawat. / International Journal of Biopharmaceutics. 2015; 6(2): 60-65.

e- ISSN 0976 - 1047

Print ISSN 2229 - 7499

IJB
International Journal of Biopharmaceutics

Journal homepage: www.ijbonline.com

EVALUATION OF SYNERGISTIC EFFECT OF GINGER, GARLIC,

TURMERIC EXTRACTS ON THE ANTIMICROBIAL ACTIVITY OF
DRUGS AGAINST BACTERIAL PATHOGENS
Seema Rawat*
Department of Botany and Microbiology, H.N.B Garhwal (Central) University, Srinagar, Uttarakhand, India.

ABSTRACT
The present work was carried out to determine the antimicrobial potential of spices and to study whether the crude
extracts of spices exhibit any synergistic effect when used in combination with antibiotics. All extracts were most effective
against E. coli. The ethanolic extract of turmeric exhibited maximum antimicrobial potential against E. coli (29.3±0.17 mm)
followed by garlic (25±0.17 mm) and ginger(21.0±0.34 mm). Amongst methanolic extracts garlic exhibited maximum
potential (15.8±0.19 mm) followed by that of ginger (15.0±0.12 mm) and turmeric (11.6±0.17 mm). The aqueous extract of
turmeric was most effective against E. coli (14.3±0.21 mm) followed by ginger (13.0±0.16 mm) while aqueous extract of
garlic showed maximum antimicrobial potential against Staphylococcus (12.6±0.13 mm). Maximum isolates were observed
to be resistant to clindamycin, oxacillin and ampicillin. The crude extracts showed synergistic effect with antibiotics in
exhibiting antimicrobial potential against bacterial pathogens. This synergistic effect can be used to design good therapeutic
approach to combat with bacterial pathogens.

Key words: Antimicrobial potential, Garlic, Ginger, Turmeric, E. coli, Staphylococcus.

INTRODUCTION
Desolate warnings issued by several health
authorities are clear indication of us standing at the
doorstep of a post-antibiotic era (CDC, 2013; WHO,
2014). Overuse of drugs has led to the evolution of drug
resistance mechanism amongst the pathogens (Kiffer et
al., 2007; AlJohani et al., 2010). The pathogens have
evolved various mechanisms to counteract the effect of
drugs. The resistance to antibiotics can be natural,
acquired, genetic, phenotypic or biological (Thomas and
Singh, 2013). The resistance may develop due to
spontaneous mutation in gene, acquisition of plasmid or
transposon, change in the physiological state of bacterial
cell or reduced permeability of cell. Major mechanisms
Corresponding Author
Seema Rawat
E-mail: seemamillenium@gmail.com
of bacterial resistance to antimicrobial agents include the
following: (a) enzymatic drug inactivation; (b) drug
target modification; (c) drug permeability reduction; and
(d) active efflux of drugs (Davies, 1994, Webber and
Piddock, 2003; Fabrega et al., 2009; Drapeau et al.,
2010). The bacteria harbouring these drugs resist,
survive, or even grow, inthe presence of a given
antimicrobial agent. Moreover, certain bacterial variants
have evolved mechanisms to resist multiple drugs,
making such variants obstinate to chemotherapy against
such bacterial strains that are the causative agents of
infection in patients. The various drug inactivation
mechanisms involve enzymatic hydrolysis of antibiotics,
group transfer, ribosome protection and biofilm
formation (Wright, 2005; Roberts, 2005; Hoiby, 2010).
There are several non-antibiotic approaches with specific
regard to antimicrobials to the treatment and prevention
of infection including probiotics, bacteriophages and

Seema Rawat. / International Journal of Biopharmaceutics. 2015; 6(2): 60-65.
phytomedicines (Aliskyet al., 1998; Caelliet al., 2000;
Fillho-Lima et al., 2000). The present study was carried
out to investigate the synergistic effect of ginger, garlic
and turmeric on the antimicrobial activity of antibiotics
against the bacterial pathogens.
MATERIALS AND METHODS
Bacterial culture
The pathogenic bacteria viz., E. coli,
Pseudomonas, Proteus, Serratia, Staphylococcus and
Klebsiella were taken from departmental culture
collection.
Acquisitions of spices& preparation of extracts
Garlic, ginger and turmericwere procured from
the local market. The spices were sorted for separation
of dirt and unwanted materials and grounded into
fine powder. Three extractantsi.e, water, ethanol and
methanol were used. The extracts were prepared by
dissolving spices in solvents in a concentration of 1:4 and
keeping at room temperature for 24hrs in a sterile beaker
covered with aluminium foil to avoid evaporation and
then subjected to filtration through sterilized Whatman
no. 1 filter paper. The solvent was dried and concentrated
using orbital shaker at 40°C. The stock solutions of the
extracts thus obtained were prepared by diluting the dried
extracts with 50% of respective solvents.
Evaluation of antimicrobial activity of extracts
The antimicrobial activity of crude extracts
against pathogenic bacteria was evaluated by using agar
well diffusion method. The isolates were inoculated into
10ml of sterile nutrient broth, and incubated at 37±1 0C
overnight. The turbidity of culture was compared with
Mac Farland standard number II. The cultures were
swabbed on the surface of sterile Mueller-Hinton agar
plates using a sterile cotton swab and allowed to dry for
3-5 minutes. Agar wells were prepared with the help
sterilized borer with 10mm diameter. The extract of
spices was diluted to give the final concentration
1000ppm, 2000ppm, 3000ppm and 4000ppm. 100 µl of
different dilutions of the extracts was added to the wells
of the inoculated plates. 50% ethanol and 50% methanol
was used as control which was introduced into the well
instead of the extract. The plates were incubated in an
upright position at 37±10C for 24hrs. The zone of
inhibition was measured and expressed in millimetres
(mm).
Antibiotic sensitivity assay
All isolates were tested for antibiotic
sensitivity by Kirby-Bauer disc diffusion method
(Bauer et al., 1996) on Mueller-Hinton agar (MHA).
The cultures were enriched in sterile nutrient Broth
overnight at 37˚C. Using a sterile cotton swabs, the
cultures were aseptically swabbed on the surface of
61
surface MHA plates and allowed to dry for 3-5
minutes before applying the antibiotic discs. Using a
sterile forcep, 4 antibiotic discs were aseptically
placed over the inoculated plates sufficiently
separated from each other to avoid overlapping of
inhibition zones. The plates were incubated in an
upright position for 24hrs at 37˚C and diameter of
zone of inhibition was measured in mm.
Evaluation of synergistic effect of crude extracts on
the antimicrobial activity of drugs
The antibiotic and dilution of the crude extracts
exhibiting maximum antimicrobial potential against the
bacterial pathogens was chosen for further study. This
test was carried out in the similar as described under
antibiotic sensitivity assay with an addition that 100 µl of
the dilutions of the extracts exhibiting maximum
antimicrobial potential was added to the antibiotic discs.
The plates were incubated in an upright position at
37±10C for 24hrs. The zone of inhibition was measured
and expressed in millimetres (mm).
RESULTS
Antimicrobial activity of crude extracts against
bacterial pathogens
All extracts exhibited good antimicrobial
potential towards uropathogens (Table 1 to 3). The
ethanolic extract of turmeric exhibited maximum
antimicrobial potential against E. coli (29.3±0.17 mm)
and least activity towards Serratia (7.3±0.14 mm). The
methanolic extract exhibited maximum activity againstE.
coli (11.6±0.17 mm) and least against Serratia (6.5±0.14
mm). The aqueous extract exhibited maximum activity
against E. coli (14.3±0.21 mm) and minimum against
Serratia (6.9±0.12 mm). The ethanolic extract of garlic
showed maximum antimicrobial potential towards E. coli
(25±0.17 mm) and least towards Klebsiella (7.3±0.18
mm). The methanolic extract was most effective against
E. coli (15.8±0.19 mm) and least towardsKlebsiella
(7.0±0.14 mm) while aqueous extract was most effective
against Staphyloccus(12.6±0.13 mm) and least towards
Klebsiella (5.6±0.14 mm). The ethanolic extract of ginger
showed maximum activity against E.coli (21.0±0.34 mm)
and least activity against Serratia (9.3±0.32 mm). The
methanolic extract was most effective against E. coli
(15.0±0.12 mm) and least against Serratia (6.3±0.12
mm). The aqueous extract was most effective against E.
coli (13.0±0.16 mm) and least against Serratia (6.0±0.12
mm).
Antimicrobial activity of antibiotics against bacterial
pathogens
Maximum isolates were observed to be resistant
to clindamycin, oxacillin and ampicillin (Fig. 1). E. coli
was found to be resistant to ciprofloxacin.
Staphylococcus was observed to be resistant
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Seema Rawat. / International Journal of Biopharmaceutics. 2015; 6(2): 60-65.

to clindamycin, erthyromycin, oxacillin, vancomycin, Fig 1. Effect of antibiotics against bacterial pathogens
ampicillin and ciprofloxacin. Pseudomonas was resistant
to erythromycin while Klebsiella was resistant to
clindamycin, chloramphenicol, oxacillin, vancomycin,
ampicillin, ciprofloxacin and cephalothin. Proteus was
resistant to clindamycin, eryhtromycin, oxacillin,
ampicillin and cephalothin. Serratia was resistant to
clindamycin, eryhtromycin, oxacillin, vancomycin,
ampicillin, ciprofloxacin and cephalothin.

Synergistic effect of crude extracts on the

antimicrobial activity of drugs
The extracts of different spices showed
synergistic effect with antibiotics in exhibiting
antimicrobial potential against bacterial pathogens (Table CLI- Clindamycin; TPM- Trimethoprim; CHL- Chloramphenicol; ERY-
4 to 6). The zone diameters were found to increase when Erythromycin; TOB- Tobramycin; OX- Oxacillin; VAN- Vancomycin;
extracts were used in combination with antibiotics. The AMP- Ampicillin; AMK- Amikacin; CIP- Ciprofloxacin, CEPH-
Cephalothin.
combination was found to be more potent than either of
the two.

Table 1a. Antimicrobial activity of ethanolic extract of turmeric against bacterial pathogens
Zone of inhibition (mm)
Name of organism
1000 ppm 2000 ppm 3000 ppm 4000 ppm
E. coli 11.6±0.15 16.3±0.25 21.3±0.14 29.3±0.17
Staphylococcus 8.6±0.14 9.3±0.15 10.7±0.20 13.6±0.13
Pseudomonas 3.6±0.10 5.3±0.12 7.4±0.20 8.6±0.20
Klebsiella 8.6±0.10 10.3±0.21 12.6±0.14 14.3±0.18
Proteus 9.3±0.12 11.6±0.12 12.6±0.22 13.3±0.21
Serratia 3.3±0.15 4.3±0.21 5.3±0.12 7.3±0.14
Values are mean ± SD of three replicates

Table 1b. Antimicrobial activity of methanolic extract of turmeric against bacterial pathogens
Zone of inhibition (mm)
Name of organism
1000 ppm 2000 ppm 3000 ppm 4000 ppm
E. coli 5.6±0.15 7.3±0.14 9.4±0.21 11.6±0.17
Staphylococcus 7.3±0.22 8.3±0.25 9.6±0.20 10.6±0.12
Pseudomonas 3.0±0.12 4.5±0.14 6.2±0.12 7.6±0.20
Klebsiella 7.4±0.10 8.6±0.12 9.0±0.12 10.6±0.24
Proteus 3.3±0.24 5.6±0.15 6.6±0.25 8.3±0.15
Serratia 2.3±0.15 3.8±0.15 4.9±0.22 6.5±0.14
Values are mean ± SD of three replicates

Table 1c. Antimicrobial activity of aqueous extract of turmeric against bacterial pathogens
Zone of inhibition (mm)
Name of organism
1000 ppm 2000 ppm 3000 ppm 4000 ppm
E. coli 6.3±0.20 8.6±0.23 10.3±0.21 14.3±0.21
Staphylococcus 7.6±0.26 8.7±0.14 9.8±0.16 11.3±0.23
Pseudomonas 3.4±0.20 4.9±0.21 6.8±0.15 8.0±0.14
Klebsiella 7.8±0.20 9.3±0.15 10.6±0.30 12.8±0.14
Proteus 4.3±0.10 6.5±0.12 8.6±0.12 10.3±0.12
Serratia 3.0±0.12 4.0±0.12 5.1±0.24 6.9±0.12
Values are mean ± SD of three replicates
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Seema Rawat. / International Journal of Biopharmaceutics. 2015; 6(2): 60-65.

Table 2a. Antimicrobial activity of ethanolic extract of garlic against bacterial pathogens
Zone of inhibition (mm)
Name of organism
1000 ppm 2000 ppm 3000 ppm 4000 ppm
E. coli 19±0.15 21±0.10 23±0.13 25±0.17
Staphylococcus 7.0±0.14 10±0.15 12±0.11 15±0.12
Pseudomonas 6.7±0.21 7.8±0.17 8.6±0.13 9.3±0.14
Klebsiella 4.0±0.12 5.4±0.16 6.7±0.12 7.3±0.18
Proteus 6.3±0.14 8.2±0.12 9.4±0.22 10.3±0.15
Serratia 3.3±0.12 5.7±0.20 7.5±0.14 9.3±0.12
Values are mean ± SD of three replicates
Table 2b. Antimicrobial activity of methanolic extract of garlic against bacterial pathogens
Zone of inhibition (mm)
Name of organism
1000 ppm 2000 ppm 3000 ppm 4000 ppm
E. coli 9.3±0.17 11.6±0.13 12.8±0.23 15.8±0.19
Staphylococcus 6.6±0.14 7.3±0.20 9.3±0.16 11.5±0.13
Pseudomonas 6.0±0.18 7.3±0.11 8.2±0.17 9.0±0.17
Klebsiella 3.3±0.17 4.8±0.20 6.3±0.12 7.0±0.14
Proteus 6.0±0.12 7.5±0.16 9.0±0.10 10.0±0.12
Serratia 3.0±0.14 4.8±0.17 6.1±0.10 8.2±0.10
Values are mean ± SD of three replicates
Table 2c. Antimicrobial activity of aqueous extract of garlic against bacterial pathogens
Zone of inhibition (mm)
Name of organism
1000 ppm 2000 ppm 3000 ppm 4000 ppm
E. coli 3.4±0.15 4.3±0.10 5.4±0.13 10.2±0.17
Staphylococcus 4.5±0.12 6.7±0.15 10.5±0.12 12.6±0.13
Pseudomonas 5.0±0.10 6.5±0.12 7.5±0.13 8.7±0.27
Klebsiella 2.2±0.17 3.4±0.14 4.1±0.12 5.6±0.14
Proteus 5.0±0.12 6.2±0.12 7.5±0.12 9.2±0.15
Serratia 2.5±0.10 3.7±0.17 4.9±0.13 6.0±0.10
Values are mean ± SD of three replicates

Table 3a. Antimicrobial activity of ethanolic extract of ginger against bacterial pathogens
Zone of inhibition (mm)
Name of organism
1000 ppm 2000 ppm 3000 ppm 4000 ppm
E. coli 17.0±0.15 18.0±0.15 20.0±0.18 21.0±0.14
Staphylococcus 9.2±0.14 10.0±0.15 12.5±0.21 14.0±0.12
Pseudomonas 10.0±0.20 14.0±0.17 16.0±0.13 18.0±0.18
Klebsiella 6.0±0.12 8.4±0.16 10.7±0.12 12.3±0.18
Proteus 10.5±0.14 11.2±0.12 13.6±0.12 15.3±0.15
Serratia 4.7±0.12 6.7±0.21 7.4±0.14 9.3±0.12
Values are mean ± SD of three replicates

Table 3b. Antimicrobial activity of methanolic extract of ginger against bacterial pathogens
Zone of inhibition (mm)
Name of organism
1000 ppm 2000 ppm 3000 ppm 4000 ppm
E. coli 11.0±0.25 12.0±0.15 14.0±0.13 15.0±0.12
Staphylococcus 8.0±0.14 9.2±0.15 10.7±0.21 11.7±0.12
Pseudomonas 8.5±0.21 10.0±0.17 11.6±0.10 12.5±0.24
Klebsiella 4.0±0.12 6.0±0.16 7.0±0.12 8.5±0.18
Proteus 6.2±0.24 7.2±0.12 8.4±0.20 9.7±0.25
Serratia 3.2±0.12 4.7±0.21 5.4±0.14 6.3±0.12
Values are mean ± SD of three replicates.
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Seema Rawat. / International Journal of Biopharmaceutics. 2015; 6(2): 60-65.

Table 3c. Antimicrobial activity of aqueous extract of ginger against bacterial pathogens
Zone of inhibition (mm)
Name of organism
1000 ppm 2000 ppm 3000 ppm 4000 ppm
E. coli 9.0±0.15 10.6±0.17 11.4±0.10 13.0±0.16
Staphylococcus 7.3±0.14 8.7±0.24 10±0.21 12±0.21
Pseudomonas 4.57±0.17 5.8±0.22 6.4±0.20 8.3±0.12
Klebsiella 3.4±0.12 5.4±0.19 6.7±0.12 7.9±0.10
Proteus 2.3±0.14 3.6±0.12 4.75±0.22 6.5±0.15
Serratia 2.7±0.12 4.3±0.14 5.1±0.20 6.0±0.12
Values are mean ± SD of three replicates.
Table 4. Antimicrobial activity of turmeric extracts in combination with antibiotics against bacterial pathogens
Antibiotic +
Zone of Antibiotic + Antibiotic +
methanolic
Name of organism Antibiotic inhibition ethanolic extract aqueous extact
extract
(mm) (mm) (mm)
(mm)
E. coli Amp 29.0±0.67 45.5±0.20 36.0±0.16 38.1±0.30
Staphylococcus Chl 24.0 ±0.35 34.4±0.50 30.2±0.32 32.2±0.30
Pseudomonas Cip 19.0 ±0.48 25.0±0.34 22.0±0.36 23.0±0.22
Klebsiella Amk 11.0±0.62 20.0 ±0.24 14.0±0.30 17.0 ±0.24
Proteus Chl 23.0±0.56 40.5±0.36 34.0±0.32 37.2±0.35
Serratia Chl 13±0.45 18.0±0.30 13.7±0.25 16.4±0.35
Amp- Ampicillin; Chl- Chlorafloxacin; Cip- Ciprofloxacin; Amk- Amikacin Conc. of extracts used: 4000 ppm.
Table 5. Antimicrobial activity of garlic extracts in combination with antibiotics against bacterial pathogens
Antibiotic + Antibiotic +
Antibiotic +
Zone of methanolic aqueous
Name of organism Antibiotic ethanolic extract
inhibition (mm) extract extact
(mm)
(mm) (mm)
E. coli Amp 29.0±0.67 46.0±0.39 36.3±0.34 31.0±0.35
Staphylococcus Chl 24.0 ±0.35 32.4±0.56 30.1±0.24 27.2±0.32
Pseudomonas Cip 19.0 ±0.48 24.2±0.40 23.0±0.30 22.4±0.24
Klebsiella Amk 11.0±0.62 18.2 ±0.34 16.4±0.24 13.2 ±0.27
Proteus Chl 23.0±0.56 30.4±0.33 27.1±0.32 20.4±0.21
Serratia Chl 13.0±0.45 19.4±0.37 16.2±0.25 15.1±0.26
Amp- Ampicillin; Chl- Chlorafloxacin; Cip- Ciprofloxacin; Amk- Amikacin; Conc. of extracts used: 4000 ppm.
Table 6. Antimicrobial activity of ginger extracts in combination with antibiotics against bacterial pathogens
Name of Zone of Antibiotic + Antibiotic + Antibiotic +
organism Antibiotic inhibition ethanolic extract methanolic extract aqueous extact
(mm) (mm) (mm) (mm)
E. coli Amp 29.0±0.67 41.0±0.24 36.3±0.16 30.0±0.35
Staphylococcus Chl 24.0 ±0.35 32.1±0.56 27.5±0.34 25.0±0.30
Pseudomonas Cip 19.0 ±0.48 30.4±0.34 26.3±0.30 20.4±0.24
Klebsiella Amk 11.0±0.62 20.5 ±0.24 17.4±0.26 15.2±0.27
Proteus Chl 23.0±0.56 33.1±0.33 30.4±0.32 26.5±0.21
Serratia Chl 13.0±0.45 19.3±0.35 17.0±0.25 14.5±0.30
Amp- Ampicillin; Chl- Chlorafloxacin; Cip- Ciprofloxacin; Amk- Amikacin Conc. of extracts used: 4000 ppm.

DISCUSSION AND CONCLUSION increasing resistance amongst pathogens towards

The spices used in cooking are well known since
ages for adding flavor, colour and aroma to the food.
They are part of our daily diet. These spices also hold
medicinal values and therefore widely used in traditional
medical practices. The spices have started gaining
attention of scientists as an alternative approach due to
antibiotics (Uraih, 2004; Souza, 2005; Pundir and Jain,
2010). They possess a number of pharmacological
effects to treat different human ailments (Arora and Kaur,
1999; Gur et al., 2006).
Many workers are now-a-days working on the
antimicrobial potential of extracts of medicinal plants,
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Seema Rawat. / International Journal of Biopharmaceutics. 2015; 6(2): 60-65.

spices and herbs however there is a need to evaluate the
synergism between these extracts and antibiotics usually
prescribed to treat infections. This may prove to be
beneficial and probably will not allow bacterial
pathogens to easily develop resistance. The present work
was focused on synergistic aspect only.
All extracts were found to be most effective
against E. coli. The ethanolic extract of turmeric
exhibited maximum antimicrobial potential as compared
to aqueous and aqueous was more potent than methanolic
extract. However in case of ginger and garlic aqueous
extract was found to be less effective as compared to
methanolic extract. The solubility of phytochemicals in
different solvents decides which will be most effective
and therefore in this study ethanolic, aqueous and
methanolic extracts were used.
The active constituent of spices may exhibit
their antimicrobial effect either by degradation of cell
wall, disruption of cytoplasmic membrane, leakage of
cellular components, damage protein, interfere with the
enzymatic activities inside cell, affect synthesis of DNA
and RNA, affect electron transport and nutrient uptake,
leakage of cellular components, impair the energy
production inside cell, change fatty acid and phospholipid
constituents (Shan et al., 2007). The extracts showed
synergistic effect with antibiotics in exhibiting
antimicrobial potential against bacterial pathogens.The
combination was found to be more potent than either of
the two. Thus it may be concluded that the combination
of antibiotics alongwith spices can be effectively used to
combat various infections.

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