Available online at www.jpsscientificpublications.

com

Life Science Archives (LSA)

ISSN: 2454-1354
Volume 1; Issue - 1; Year 2015; Page: 13- 27

Research Article
SECLUSION AND MICROBIAL DEPICTION OF MULTIPLE DRUG RESISTANT
Salmonella spp. FROM TAMILNADU
Balasubramani Gangathraprabhu1, and Ponnusamy Ponmurugan*2,
1

Kodai Darwin Institute of Research and Technology, Kodaikanal, Tamil Nadu, India.
Department of Biotechnology, K.S.R College of Technology, Tiruchengode, Tamil Nadu, India.
Abstract
2

Salmonellosis continues to be a major public health problem worldwide. It also contributes to

negative economic impacts due to the cost of surveillance investigation, treatment and prevention of illness.
As such, research on Salmonella has gained great interest and concern from scientists. The purpose of this
study is to discuss the classification and characteristic, clinical manifestation, epidemiology, antibiotic
resistance and quorum sensing of Salmonella. Salmonella is closely related to the Escherichia genus and are
found worldwide in cold- and warm-blooded animals (including humans), and in the environment. They
cause illnesses like typhoid fever, paratyphoid fever, and food borne illness. In the present study, Multiple
Drug Resistant (MDR) Salmonella typhi was isolated from various geographical areas and confirmed its
antibiotic pattern by Kirby-Bauer principle.
Article History
Received : 04.02.2015
Revised : 16.02.2015
Accepted : 25.02.2015
1. Introduction

Key words: Multiple drugs resistant,

Salmonella.spp. Antibiogram and Antibiotic
sensitivity

There are 16 million annual cases of

typhoid fever, 1.3 billion cases of gastroenteritis
and 3 million deaths worldwide due to Salmonella
(Bhunia, 2008). Salmonella, a Gram-negative
bacilli member of the family enterobacteriaceae,
ranging approximately 2-3 0.4 -0.6 m in size is
a causative agent of enteric (typhoid) fever and
gastroenteritis. Over 2000 serotypes of Salmonella
infects human and virtually all known wild and
domestic animals, including birds, reptiles, and
insects. It is among the most commonly isolated
foodborne pathogens associated with fresh fruits
and vegetables. In recent years, the incidence of
foodborne outbreaks caused by the contamination
of fresh fruits and vegetables has increased and

*Corresponding author: Ponnusamy Ponmurugan

Tel.: +91 9788273734
E-mail: gprabhubt@gmail.com

become a great concern in industrialized countries.

Outbreaks of salmonellosis have been linked to a
wide variety of fresh fruits and vegetables
including apple, cantaloupe, alfalfa sprout, mango,
lettuce, cilantro, unpasteurized orange juice,
tomato, melon, celery and parsley (Pui et al.,
2011). Nowadays, Salmonella continued to
develop resistance against most antibiotics
(Baggesen and Wegener, 1994; Baggesen and
Aarestrup, 1998; Molbak et al., 1999).
Multi-drug resistant S. typhi were first
described in India in 1990 (Jesudasan et al., 1990)
and several different phage types carrying incH1
plasmids of 110-120 MDa have been described
(Threlfall et al., 1992). To better monitor and
control the spread of MDR Salmonella, it is
important to understand the mechanisms
responsible for drug resistance and how drug
resistance is transmitted to and between

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015

Salmonella strains. This present research

summarizes current knowledge on antimicrobial
drugs used to treat Salmonella infections and
provides an overview of MDR Salmonella in the
United States and a discussion of the genetics of
Salmonella drug resistance, including the
mechanisms responsible for the transmission of
drug-resistance genes in Salmonella, using data
from the United States and other countries
(Alcaine et al., 2007).
The increase of multidrug-resistant pathogens
of human and animal origins is a major public
health concern. For a better understanding of the
health consequences of multidrug-resistant
bacteria transmitted from animal products to
humans, the host interaction of zoonotic
Salmonella isolates along with other pathogenic
and commensally bacteria was evaluated using a
human intestinal Caco-2 cell system. Multidrugresistant S. agona, S. heidelberg and
S. typhimurium possessed plasmid mediated class
1 integrons. S. typhimurium DT104 isolate from
ground beef showed the well-known genotypic
and phenotypic resistance characteristics of the
species, and contained the chromosomally located
class 1 integron. Among the multidrug- resistant
Salmonella isolates, the S. heidelberg 219 had the
highest invasion number at 1.0 104 CFU/mL,
followed by the S. typhimurium DT104 isolate at
7.7 10(3) CFU/mL. Listeria monocytogenes was
the best performed among the tested species in
invading the Caco-2 cell. Multidrug-resistant
opportunistic pathogens Klebsiella pneumoniae
and Pseudomonas aeruginosa were also able to
invade the cells. The invasion of S. heidelberg
219,
S.
typhimurium
DT104
and
L. monocytogenes, the bacterial counts increased 2
log cycles in 9 hrs in the Caco-2 cells. Therefore,
these strains could rapidly proliferate after their
invasion into the cells (Kim et al., 2007).
2. Methodology
2.1. Isolation and characterization of
Salmonella strains from various geographical
areas
A total of one hundred and thirty samples
were collected from five different sources and five
different regions in and around Tamilnadu under

14

aseptic conditions and transported to the

laboratory as soon as possible for bacteriological
examination especially for the presence of
Salmonella spp. Each sample was serially diluted
and inoculated in Salmonella Shigella agar,
MacConkey agar medium separately and further
each strain was identified as per the standard
procedure of Bergeys manual of determinative
bacteriology.
2.2. Antibiogram of Salmonella Isolates using
Antimicrobial Susceptibility Test
Antibiotic susceptibility test of the
organism was carried out for eleven antibiotics.
Antimicrobial susceptibility tests were performed
by Kirby-Bauer disc diffusion method. Overnight
cultures in peptone water were lawned over the
Mueller-Hinton agar (Hi-media). The antibiotic
discs were used at the following concentration:
Ampicillin (A) 30 mcg, Amikacin (Ak) 30 mcg,
Amoxicillin (Am) 25 mcg, Chloramphenicol (C)
25 mcg, Ciprofloxacin (Cf) 30 mcg, Cotrimaxazole (Co) 25 mcg, Gentamicin (G) 30 mcg,
Nalidixic acid (Na) 30 mcg, Oflaxacin (O) 30
mcg, Rifampicin (R) 30 mcg, Tetracycline (T) 10
mcg (Hi-media).
3. Result
3.1. Collection of samples from various
geographical areas for Salmonella spp.
The present study identified Erode,
Namakkal, Perundurai, Karur and Salem as
poultry units concentrated areas in the northern
Tamilnadu, India. 27 chicken flesh samples were
collected from Erode, 35 poultry waste samples
were collected from Namakkal, 20 contaminated
food samples from Perundurai, 23 Deceased
chicken samples from Salem and 25 contaminated
egg samples were collected from Karur districts
(Table - 1).
3.2.
Microbes
isolated
Geographical area

from

various

Out of 130 samples collected from five

different districts of northern Tamilnadu only 87
isolates were found to be Salmonella spp. Highest
number of Salmonella spp. were isolated from
samples collected from Namakkal (26 isolates)
which are poultry wastes, followed by 19 isolates

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015

from fresh chicken flesh samples collected from

Erode and 17 isolates from contaminated egg
samples of Karur. Least number of Salmonella
isolates was found to be in contaminated food
samples collected from Perundurai isolates (11
numbers). However, other microorganisms were
also recorded in the samples mainly Shigella and
fungal species (Table - 2).
3.3. Antibiogram of Salmonella Isolates
Antibiotic sensitivity test were performed
for the isolates using Kirby method Kirby-Bauer
disc diffusion method. An overnight culture in
peptone water was lawned over the MuellerHinton agar (Hi-media).
The commercially available antibiotic
discs used in the following concentration:
ampicillin (A) 30 mcg, amikacin (Ak) 30 mcg,
amoxicillin (Am) 25 mcg, chloramphenicol (C) 25
mcg, ciproflaxacin (Cf) 30 mcg, co-trimaxazole

15

(Co) 25 mcg, gentamicin (G) 30 mcg, nalidixic

acid (Na) 30 mcg, oflaxacin (O) 30 mcg,
rifampicin (R) 30 mcg, tetracycline (T) 10 mcg
(Hi-media). Results showed zone of inhibition in
the range of 9 to 20 mm, large zone of inhibition
was 20 mm for Ciproflaxin, followed by 18 mm
zone of inhibition for Oflaxin and 16 mm zone of
inhibition for Co-trimaxazole. Smallest zone of
inhibition was 9 mm for Tetracycline, followed by
10 mm for Amicacin and Amoxicillin, 11 mm
zone of inhibition for Ampicillin, Gentamycin and
Rifampicin, 12 mm and 13 mm zone of inhibition
for Nalidixic acid and Chlorampenicol
respectively. As per the standards, isolates with
large zone of inhibition were identified as
sensitive to the respective antibiotics (Table 3A
to 3I). Antibiogram of Best MDR Salmonella
isolates were picked up (Table - 4).

Table - 1: Collection of Samples from various geographical areas for Salmonella spp.
S.
No.
1.
2.
3.
4.
5.

Place

Number
of samples
Erode
27
Namakkal
35
Perundurai
20
Salem
23
Karur
25

Source
Fresh Chicken Flesh
Poultry waste
Contaminated Food
Deceased chicken
Contaminated Egg

Table - 2: Microbes isolated from various Geographical area

S.
No
1.
2.
3.
4.
5.

Geographical Collected
Area
samples
Erode
27
Namakkal
35
Perundurai
20
Salem
23
Karur
25
Total
130

Number of
Salmonella spp.
19
26
11
14
17
87

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015

16

Sa-5

Sa-6

Sa-7

Sa-8

Sa-9

10

Sa-10

S
(22mm)
S
(18mm)
S
(25mm)
R
(13mm)
S
(23mm)
S
(17mm)
S
(19mm)
S
(22mm)
S
(27mm)
S
(20mm)

S
(19mm)
S
(22mm)
S
(19mm)
R
(11mm)
S
(20mm)
S
(24mm)
S
(25mm)
S
(20mm)
S
(20mm)
S
(26mm)

S
(18mm)
S
(20mm)
S
(26mm)
R
(13mm)
S
(21mm)
S
(19mm)
S
(20mm)
S
(19mm)
S
(23mm)
S
(26mm)

S
(17mm)
S
(19mm)
S
(15mm)
R
(12mm)
S
(18mm)
S
(16mm)
S
(15mm)
S
(21mm)
S
(19mm)
S
(22mm)

S
(21mm)
S
(26mm)
S
(20mm)
M
(14mm)
S
(26mm)
S
(25mm)
S
(22mm)
S
(24mm)
S
(20mm)
S
(26mm)

S
(16mm)
S
(21mm)
S
(19mm)
S
(18mm)
S
(19mm)
S
(17mm)
S
(21mm)
S
(18mm)
S
(16mm)
S
(16mm)

S
(21mm)
S
(18mm)
S
(15mm)
M
(14mm)
S
(16mm)
S
(20mm)
S
(24mm)
S
(18mm)
S
(16mm)
S
(15mm)

S
(19mm)
S
(22mm)
S
(26mm)
R
(13mm)
S
(21mm)
S
(23mm)
S
(19mm)
S
(20mm)
S
(22mm)
S
(24mm)

S
(16mm)
S
(20mm)
S
(17mm)
S
(19mm)
S
(19mm)
S
(22mm)
S
(18mm)
S
(16mm)
S
(16mm)
S
(19mm)

S
(20mm)
S
(22mm)
S
(19mm)
R
(16mm)
S
(21mm)
S
(24mm)
S
(19mm)
S
(22mm)
S
(26mm)
S
(21mm)

S
(23mm)
S
(20mm)
S
(23mm)
S
(22mm)
S
(19mm)
S
(24mm)
S
(24mm)
S
(20mm)
S
(19mm)
S
(19mm)

Multi Drug
Resistant

Tetracycline
10(g)/disc

Sa-4

Rifampicin
30(g)/disc

Oflaxacin
30(g)/disc

Sa-3

Nalidixic acid
30(g)/disc

Gentamicin
30(g)/disc

Sa-2

Co-trimaxazole
25(g)/disc

Ciproflaxacin
30(g)/disc

Sa-1

Chloramphenicol
25(g)/disc

Amoxicillin
25(g)/disc

Strain
No.

Amikacin
30(g)/disc

S. No

Ampicillin
30(g)/disc

Table 3(A): Antibiogram pattern of Salmonella strains

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015

17

Sa-15

Sa-16

Sa-17

Sa-18

Sa-19

10

Sa-20

Multi Drug
Resistant

Tetracycline
10(g)/disc

Sa-14

Rifampicin
30(g)/disc

Oflaxacin
30(g)/disc

Sa-13

Nalidixic acid
30(g)/disc

Gentamicin
30(g)/disc

Sa-12

Co-trimaxazole
25(g)/disc

Ciproflaxacin
30(g)/disc

Sa-11

Chloramphenicol
25(g)/disc

Amoxicillin
25(g)/disc

Strain
No.

Amikacin
30(g)/disc

S. No

Ampicillin
30(g)/disc

Table 3(B): Antibiogram pattern of Salmonella strains

S
(26mm)
S
(20mm)
S
(19mm)
S
(22mm)
S
(27mm)
R
(12mm)
S
(19mm)
S
(17mm)
S
(26mm)
S
(20mm)

S
(17mm)
S
(18mm)
S
(20mm)
S
(26mm)
S
(18mm)
R
(13mm)
S
(20mm)
S
(21mm)
S
(20mm)
S
(18mm)

S
(19mm)
S
(20mm)
S
(23mm)
S
(21mm)
S
(23mm)
R
(13mm)
S
(22mm)
S
(20mm)
S
(19mm)
S
(23mm)

S
(17mm)
S
(19mm)
S
(15mm)
S
(19mm)
S
(18mm)
R
(12mm)
S
(15mm)
S
(21mm)
S
(19mm)
S
(22mm)

S
(22mm)
S
(20mm)
S
(26mm)
S
(21mm)
S
(24mm)
S
(21mm)
S
(25mm)
S
(22mm)
S
(21mm)
S
(26mm)

S
(26mm)
S
(22mm)
S
(19mm)
S
(16mm)
S
(17mm)
S
(19mm)
S
(21mm)
S
(19mm)
S
(16mm)
S
(16mm)

S
(23mm)
S
(18mm)
S
(20mm)
S
(19mm)
S
(18mm)
R
(12mm)
S
(16mm)
S
(23mm)
S
(19mm)
S
(23mm)

S
(20mm)
S
(22mm)
S
(19mm)
S
(21mm)
S
(26mm)
S
(22mm)
S
(25mm)
S
(25mm)
S
(24mm)
S
(26mm)

S
(17mm)
S
(20mm)
S
(18mm)
S
(19mm)
S
(21mm)
M
(16mm)
S
(18mm)
S
(19mm)
S
(21mm)
S
(27mm)

S
(21mm)
S
(22mm)
S
(24mm)
S
(26mm)
S
(19mm)
S
(21mm)
S
(24mm)
S
(25mm)
S
(19mm)
S
(22mm)

S
(21mm)
S
(20mm)
S
(25mm)
S
(21mm)
S
(20mm)
R
(14mm)
S
(24mm)
S
(22mm)
S
(23mm)
S
(20mm)

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Sa-25

Sa-26

Sa-27

Sa-28

Sa-29

10

Sa-30

S
(19mm)
S
(25mm)
S
(27mm)
S
(20mm)
R
(12mm)
S
(22mm)
S
(24mm)
S
(22mm)
R
(11mm)
S
(23mm)

S
(19mm)
S
(20mm)
S
(19mm)
S
(17mm)
R
(14mm)
S
(20mm)
S
(19mm)
S
(20mm)
R
(10mm)
S
(19mm)

S
(26mm)
S
(21mm)
S
(18mm)
S
(26mm)
R
(11mm)
S
(26mm)
S
(20mm)
S
(27mm)
R
(10mm)
S
(17mm)

S
(17mm)
S
(19mm)
S
(20mm)
S
(19mm)
R
(10mm)
S
(19mm)
S
(21mm)
S
(21mm)
R
(12mm)
S
(22mm)

S
(21mm)
S
(26mm)
S
(22mm)
S
(26mm)
S
(23mm)
S
(25mm)
S
(21mm)
S
(24mm)
S
(20mm)
S
(25mm)

S
(16mm)
S
(21mm)
S
(16mm)
S
(18mm)
S
(19mm)
S
(21mm)
S
(24mm)
S
(17mm)
S
(19mm)
S
(18mm)

S
(23mm)
S
(18mm)
S
(19mm)
S
(22mm)
S
(16mm)
S
(19mm)
S
(20mm)
S
(25mm)
R
(11mm)
S
(25mm)

S
(19mm)
S
(22mm)
S
(27mm)
S
(25mm)
R
(13mm)
S
(19mm)
S
(28mm)
S
(25mm)
R
(12mm)
S
(27mm)

S
(18mm)
S
(20mm)
S
(17mm)
S
(19mm)
S
(22mm)
S
(21mm)
S
(18mm)
S
(19mm)
S
(18mm)
S
(19mm)

S
(20mm)
S
(22mm)
S
(28mm)
S
(21mm)
R
(16mm)
S
(25mm)
S
(23mm)
S
(22mm)
R
(11mm)
S
(20mm)

S
(23mm)
S
(20mm)
S
(22mm)
S
(25mm)
M
(17mm)
S
(24mm)
S
(26mm)
S
(21mm)
R
(9mm)
S
(24mm)

Multi Drug
Resistant

Tetracycline
10(g)/disc

Sa-24

Rifampicin
30(g)/disc

Oflaxacin
30(g)/disc

Sa-23

Nalidixic acid
30(g)/disc

Gentamicin
30(g)/disc

Sa-22

Co-trimaxazole
25(g)/disc

Ciproflaxacin
30(g)/disc

Sa-21

Chloramphenicol
25(g)/disc

Amoxicillin
25(g)/disc

Strain
No.

Amikacin
30(g)/disc

S. No

Ampicillin
30(g)/disc

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015
18
Table 3(C): Antibiogram pattern of Salmonella strains

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Sa-35

Sa-36

Sa-37

Sa-38

Sa-39

10

Sa-40

S
(20mm)
R
(12mm)
S
(26mm)
S
(17mm)
R
(13mm)
S
(22mm)
S
(22mm)
R
(13mm)
S
(24mm)
S
(19mm)

S
(17mm)
R
(13mm)
S
(20mm)
S
(26mm)
R
(11mm)
S
(20mm)
S
(21mm)
R
(14mm)
S
(20mm)
S
(27mm)

S
(20mm)
R
(13mm)
S
(21mm)
S
(25mm)
R
(13mm)
S
(21mm)
S
(20mm)
R
(11mm)
S
(25mm)
S
(20mm)

S
(17mm)
R
(11mm)
S
(15mm)
S
(19mm)
R
(12mm)
S
(16mm)
S
(15mm)
R
(12mm)
S
(19mm)
S
(22mm)

S
(21mm)
R
(15mm)
S
(24mm)
S
(22mm)
S
(22mm)
S
(21mm)
S
(24mm)
S
(23mm)
S
(25mm)
S
(26mm)

S
(17mm)
S
(24mm)
S
(21mm)
S
(19mm)
M
(18mm)
S
(17mm)
S
(21mm)
S
(19mm)
S
(16mm)
S
(19mm)

S
(21mm)
M
(13mm)
S
(18mm)
S
(20mm)
S
(15mm)
S
(15mm)
S
(18mm)
R
(12mm)
S
(23mm)
S
(16mm)

S
(19mm)
M
(16mm)
S
(20mm)
S
(24mm)
S
(23mm)
S
(21mm)
S
(25mm)
R
(13mm)
S
(23mm)
S
(22mm)

S
(16mm)
S
(20mm)
S
(16mm)
S
(17mm)
S
(19mm)
S
(22mm)
S
(19mm)
S
(19mm)
S
(21mm)
S
(19mm)

S
(26mm)
S
(22mm)
S
(24mm)
S
(19mm)
R
(15mm)
S
(20mm)
S
(22mm)
S
(22mm)
S
(25mm)
S
(25mm)

S
(20mm)
S
(25mm)
S
(20mm)
S
(25mm)
S
(20mm)
S
(24mm)
S
(21mm)
M
(18mm)
S
(26mm)
S
(19mm)

Multi Drug
Resistant

Tetracycline
10(g)/disc

Sa-34

Rifampicin
30(g)/disc

Oflaxacin
30(g)/disc

Sa-33

Nalidixic acid
30(g)/disc

Gentamicin
30(g)/disc

Sa-32

Co-trimaxazole
25(g)/disc

Ciproflaxacin
30(g)/disc

Sa-31

Chloramphenicol
25(g)/disc

Amoxicillin
25(g)/disc

Strain
No.

Amikacin
30(g)/disc

S. No

Ampicillin
30(g)/disc

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015
19
Table 3(D): Antibiogram pattern of Salmonella strains

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Sa-45

Sa-46

Sa-47

Sa-48

Sa-49

10

Sa-50

S
(21mm)
S
(22mm)
S
(18mm)
S
(22mm)
S
(24mm)
S
(17mm)
S
(20mm)
R
(13mm)
S
(21mm)
S
(22mm)

S
(19mm)
S
(20mm)
S
(21mm)
S
(25mm)
S
(21mm)
S
(22mm)
S
(26mm)
R
(11mm)
S
(20mm)
S
(23mm)

S
(22mm)
S
(26mm)
S
(20mm)
S
(26mm)
S
(24mm)
S
(19mm)
S
(17mm)
R
(13mm)
S
(20mm)
S
(19mm)

S
(17mm)
S
(19mm)
S
(15mm)
S
(19mm)
S
(18mm)
S
(16mm)
S
(15mm)
R
(11mm)
S
(19mm)
S
(22mm)

S
(21mm)
S
(23mm)
S
(24mm)
S
(21mm)
S
(24mm)
S
(25mm)
S
(26mm)
S
(21mm)
S
(25mm)
S
(24mm)

S
(26mm)
S
(22mm)
S
(17mm)
S
(19mm)
S
(21mm)
S
(17mm)
S
(23mm)
S
(19mm)
S
(16mm)
S
(19mm)

S
(24mm)
S
(18mm)
S
(19mm)
S
(20mm)
S
(21mm)
S
(18mm)
S
(16mm)
R
(11mm)
S
(17mm)
S
(21mm)

S
(21mm)
S
(22mm)
S
(19mm)
S
(23mm)
S
(27mm)
S
(23mm)
S
(27mm)
R
(13mm)
S
(20mm)
S
(25mm)

S
(21mm)
S
(20mm)
S
(17mm)
S
(19mm)
S
(21mm)
S
(22mm)
S
(18mm)
S
(25mm)
S
(16mm)
S
(19mm)

S
(20mm)
S
(22mm)
S
(19mm)
S
(21mm)
S
(24mm)
S
(20mm)
S
(27mm)
S
(23mm)
S
(23mm)
S
(25mm)

S
(23mm)
S
(22mm)
S
(25mm)
S
(20mm)
S
(22mm)
S
(26mm)
S
(27mm)
M
(18mm)
S
(25mm)
S
(25mm)

Multi Drug
Resistant

Tetracycline
10(g)/disc

Sa-44

Rifampicin
30(g)/disc

Oflaxacin
30(g)/disc

Sa-43

Nalidixic acid
30(g)/disc

Gentamicin
30(g)/disc

Sa-42

Co-trimaxazole
25(g)/disc

Ciproflaxacin
30(g)/disc

Sa-41

Chloramphenicol
25(g)/disc

Amoxicillin
25(g)/disc

Strain
No.

Amikacin
30(g)/disc

S.No

Ampicillin
30(g)/disc

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015
20
Table 3(E): Antibiogram pattern of Salmonella strains

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Sa-55

Sa-56

Sa-57

Sa-58

Sa-59

10

Sa-60

Multi Drug
Resistant

Tetracycline
10(g)/disc

Sa-54

Rifampicin
30(g)/disc

Oflaxacin
30(g)/disc

Sa-53

Nalidixic acid
30(g)/disc

Gentamicin
30(g)/disc

Sa-52

Co-trimaxazole
25(g)/disc

Ciproflaxacin
30(g)/disc

Sa-51

Chloramphenicol
25(g)/disc

Amoxicillin
25(g)/disc

Strain
No.

Amikacin
30(g)/disc

S. No

Ampicillin
30(g)/disc

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015
21
Table 3(F): Antibiogram pattern of Salmonella strains

S
(17mm)
S
(22mm)
S
(20mm)
S
(19mm)
S
(26mm)
R
(13mm)
S
(24mm)
S
(22mm)
S
(17mm)
S
(27mm)

S
(17mm)
S
(20mm)
S
(22mm)
S
(25mm)
S
(20mm)
R
(11mm)
S
(20mm)
S
(17mm)
S
(20mm)
S
(22mm)

S
(27mm)
S
(17mm)
S
(19mm)
S
(20mm)
S
(24mm)
R
(11mm)
S
(19mm)
S
(20mm)
S
(26mm)
S
(19mm)

S
(17mm)
S
(19mm)
S
(20mm)
S
(19mm)
S
(18mm)
R
(11mm)
S
(15mm)
S
(21mm)
S
(19mm)
S
(22mm)

S
(22mm)
S
(21mm)
S
(24mm)
S
(27mm)
S
(25mm)
S
(23mm)
S
(26mm)
S
(21mm)
S
(22mm)
S
(21mm)

S
(17mm)
S
(21mm)
S
(18mm)
S
(19mm)
S
(16mm)
S
(17mm)
S
(23mm)
S
(19mm)
S
(19mm)
S
(20mm)

S
(27mm)
S
(18mm)
S
(25mm)
S
(19mm)
S
(21mm)
M
(12mm)
S
(16mm)
S
(20mm)
S
(18mm)
S
(22mm)

S
(21mm)
S
(22mm)
S
(19mm)
S
(25mm)
S
(25mm)
R
(13mm)
S
(27mm)
S
(24mm)
S
(24mm)
S
(21mm)

S
(21mm)
S
(20mm)
S
(17mm)
S
(18mm)
S
(19mm)
S
(22mm)
S
(18mm)
S
(16mm)
S
(16mm)
S
(19mm)

S
(21mm)
S
(22mm)
S
(24mm)
S
(25mm)
S
(27mm)
S
(20mm)
S
(25mm)
S
(22mm)
S
(24mm)
S
(20mm)

S
(23mm)
S
(20mm)
S
(19mm)
S
(21mm)
S
(25mm)
S
(25mm)
S
(26mm)
S
(21mm)
S
(28mm)
S
(23mm)

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Sa-65

Sa-66

Sa-67

Sa-68

Sa-69

10

Sa-70

Multi Drug
Resistant

Tetracycline
10(g)/disc

Sa-64

Rifampicin
30(g)/disc

Oflaxacin
30(g)/disc

Sa-63

Nalidixic acid
30(g)/disc

Gentamicin
30(g)/disc

Sa-62

Co-trimaxazole
25(g)/disc

Ciproflaxacin
30(g)/disc

Sa-61

Chloramphenicol
25(g)/disc

Amoxicillin
25(g)/disc

Strain
No.

Amikacin
30(g)/disc

S. No

Ampicillin
30(g)/disc

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015
22
Table 3(G): Antibiogram pattern of Salmonella strains

S
(24mm)
S
(20mm)
S
(19mm)
S
(17mm)
S
(22mm)
R
(12mm)
S
(22mm)
S
(20mm)
S
(22mm)
S
(26mm)

S
(19mm)
S
(22mm)
S
(20mm)
S
(23mm)
S
(20mm)
R
(14mm)
S
(20mm)
S
(21mm)
S
(26mm)
S
(20mm)

S
(18mm)
S
(20mm)
S
(26mm)
S
(21mm)
S
(24mm)
R
(11mm)
S
(23mm)
S
(20mm)
S
(26mm)
S
(22mm)

S
(17mm)
S
(19mm)
S
(15mm)
S
(19mm)
S
(18mm)
R
(12mm)
S
(15mm)
S
(21mm)
S
(19mm)
S
(22mm)

S
(22mm)
S
(21mm)
S
(24mm)
S
(25mm)
S
(23mm)
M
(17mm)
S
(24mm)
S
(21mm)
S
(25mm)
S
(26mm)

S
(22mm)
S
(21mm)
S
(19mm)
S
(16mm)
S
(19mm)
R
(22mm)
S
(24mm)
S
(18mm)
S
(16mm)
S
(17mm)

S
(23mm)
S
(18mm)
S
(21mm)
S
(19mm)
S
(20mm)
R
(12mm)
S
(19mm)
S
(17mm)
S
(16mm)
S
(18mm)

S
(26mm)
S
(22mm)
S
(25mm)
S
(21mm)
S
(23mm)
M
(15mm)
S
(24mm)
S
(22mm)
S
(20mm)
S
(22mm)

S
(19mm)
S
(20mm)
S
(18mm)
S
(17mm)
S
(19mm)
S
(24mm)
S
(18mm)
S
(16mm)
S
(23mm)
S
(19mm)

S
(20mm)
S
(22mm)
S
(19mm)
S
(25mm)
S
(20mm)
S
(25mm)
S
(20mm)
S
(20mm)
S
(24mm)
S
(22mm)

S
(22mm)
S
(20mm)
S
(22mm)
S
(24mm)
S
(25mm)
S
(22mm)
S
(23mm)
S
(19mm)
S
(19mm)
S
(27mm)

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Sa-75

Sa-76

Sa-77

Sa-78

Sa-79

10

Sa-80

Multi Drug
Resistant

Tetracycline
10(g)/disc

Sa-74

Rifampicin
30(g)/disc

Oflaxacin
30(g)/disc

Sa-73

Nalidixic acid
30(g)/disc

Gentamicin
30(g)/disc

Sa-72

Co-trimaxazole
25(g)/disc

Ciproflaxacin
30(g)/disc

Sa-71

Chloramphenicol
25(g)/disc

Amoxicillin
25(g)/disc

Strain
No.

Amikacin
30(g)/disc

S. No

Ampicillin
30(g)/disc

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015
23
Table 3(H): Antibiogram pattern of Salmonella strains

S
(27mm)
S
(22mm)
S
(25mm)
S
(24mm)
S
(21mm)
R
(12mm)
S
(20mm)
R
(13mm)
S
(24mm)
S
(22mm)

S
(19mm)
S
(20mm)
S
(22mm)
S
(19mm)
S
(23mm)
R
(14mm)
S
(24mm)
R
(11mm)
S
(20mm)
S
(26mm)

S
(19mm)
S
(22mm)
S
(20mm)
S
(25mm)
S
(26mm)
R
(11mm)
S
(20mm)
R
(13mm)
S
(23mm)
S
(20mm)

S
(17mm)
S
(19mm)
S
(15mm)
S
(19mm)
S
(18mm)
R
(12mm)
S
(15mm)
R
(11mm)
S
(19mm)
S
(22mm)

S
(23mm)
S
(21mm)
S
(24mm)
S
(25mm)
S
(23mm)
S
(24mm)
S
(21mm)
S
(21mm)
S
(25mm)
S
(23mm)

S
(26mm)
S
(21mm)
S
(19mm)
S
(18mm)
S
(20mm)
S
(17mm)
S
(27mm)
S
(19mm)
S
(16mm)
S
(16mm)

S
(21mm)
S
(18mm)
S
(23mm)
S
(19mm)
S
(16mm)
R
(12mm)
S
(21mm)
S
(18mm)
S
(19mm)
S
(21mm)

S
(20mm)
S
(22mm)
S
(28mm)
S
(21mm)
S
(23mm)
S
(22mm)
S
(24mm)
R
(13mm)
S
(22mm)
S
(25mm)

S
(16mm)
S
(21mm)
S
(18mm)
S
(17mm)
S
(19mm)
S
(25mm)
S
(22mm)
S
(16mm)
S
(22mm)
S
(19mm)

S
(21mm)
S
(22mm)
S
(20mm)
S
(19mm)
S
(25mm)
M
(18mm)
S
(25mm)
S
(19mm)
S
(22mm)
S
(23mm)

S
(23mm)
S
(20mm)
S
(23mm)
S
(21mm)
S
(25mm)
R
(14mm)
S
(23mm)
M
(17mm)
S
(20mm)
S
(27mm)

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Sa-85

Sa-86

Sa-87

S
(22mm)
S
(18mm)
S
(26mm)
S
(25mm)
S
(22mm)
S
(22mm)
R
(13mm)

S
(26mm)
S
(20mm)
S
(22mm)
S
(17mm)
S
(25mm)
S
(24mm)
R
(11mm)

S
(19mm)
S
(22mm)
S
(20mm)
S
(25mm)
S
(23mm)
S
(20mm)
R
(11mm)

S
(17mm)
S
(19mm)
S
(15mm)
S
(19mm)
S
(18mm)
S
(16mm)
R
(12mm)

S
(24mm)
S
(21mm)
S
(22mm)
S
(25mm)
S
(25mm)
S
(24mm)
M
(19mm)

S
(19mm)
S
(25mm)
S
(17mm)
S
(18mm)
S
(19mm)
S
(22mm)
M
(15mm)

S
(24mm)
S
(18mm)
S
(19mm)
S
(21mm)
S
(16mm)
S
(18mm)
R
(12mm)

S
(19mm)
S
(22mm)
S
(26mm)
S
(23mm)
S
(20)
S
(26mm)
R
(12mm)

S
(17mm)
S
(21mm)
S
(17mm)
S
(17mm)
S
(20mm)
S
(22mm)
S
(18mm)

S
(21mm)
S
(23mm)
S
(22mm)
S
(25mm)
S
(26mm)
S
(27mm)
S
(22mm)

S
(27mm)
S
(22mm)
S
(21mm)
S
(21mm)
S
(27mm)
S
(20mm)
S
(25mm)

Multi Drug
Resistant

Tetracycline
10(g)/disc

Sa-84

Rifampicin
30(g)/disc

Oflaxacin
30(g)/disc

Sa-83

Nalidixic acid
30(g)/disc

Gentamicin
30(g)/disc

Sa-82

Co-trimaxazole
25(g)/disc

Ciproflaxacin
30(g)/disc

Sa-81

Chloramphenicol
25(g)/disc

Amoxicillin
25(g)/disc

Strain
No.

Amikacin
30(g)/disc

S. No

Ampicillin
30(g)/disc

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015
24
Table 3(I): Antibiogram pattern of Salmonella strains

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015

25

Table - 4: Antibiogram of Best MDR Salmonella Isolates

S.No

Antibiotic

Concentration
(mcg)/disc
30
30

Zone of
inhibition (mm)
11
10

Result

10
12
20
16
11
12

R
R
S
S
R
R

1
2

Ampicillin
Amikacin

3
4
5
6
7
8

Amoxicillin
Chloramphenicol
Ciproflaxacin
Co-Trimaxazole
Gentamicin
Nalidixic acid

25
25
30
25
30
30

9
10
11

Ofloxacin
Rifampicin
Tetracycline

30
30
10

4. Discussion
Enteric fever caused by S. typhi has been
uncontrolled problem in developing world due to
its high virulence. It has been reported that S. typhi
is resistant to two or more antibiotic among the
twelve commonly used antibiotics of the day,
importantly the Chloramphenicol, Ampicillin and
Cotrimaxole. In the present study showed that the
isolated strain was susceptible for only three
antibiotics Ciproflaxacin, Co-trimaxazole and
Oflaxacin out of eleven commonly used
antibiotics which includes the Chloramphenicol
and Ampicillin. However, it could detected that
continuous use of antibiotics for treatment further
increases its spectrum of drug resistance, which is
evident through that in the present study the
isolates are resistant to Tetracycline and
Ciprofloxacin which were once susceptible.
Therefore there is an urgent need of novel therapy
which could face this mutant strains at genomic
level. Hence, the present study was initiated to
repress the Salmonella virulence by novel gene
therapeutic approach.
Virulence is controlled at transcriptional
level by several proteins like RpoS sigma factor
(Fang et al., 1992), the cyclic AMP-binding
protein and the two components of regulatory

R
R

18
S
11
R
9
R
systems OmpR-EnvZ (Dorman et al., 1989) and
PhoP/PhpQ (Galan and Curtiss, 1989). PhoQ was
predicted
as
phosphorylating
and
dephosphorylating agent in response to
environmental changes were mutation in
PhoP/PhoQ as well as constitutive allele mapping
to PhoQ result in attenuation for virulence (Fields
et al., 1986; Miller, and Mekalanos, 1990).
Positively charged amino acids help in the
interaction of phoP/ phoQ DNA binding Proteins
with DNA.
The results of Gel mobility shift assay
confirmed that DEPC modifies the histidines
present in the binding sites of phoP/ phoQ DNA
binding Proteins. Histidine modified phoP/ phoQ
DNA binding proteins fails to interact with DNA
which may result in unexpression of virulence
genes (Gangathraprabhu et al., 2012). Thus, this
mechanism at a molecular level might be paving a
path in the treatment of enteric fever without
leading to the further MDR Salmonella strains.
5. Reference
1) Baggesen, D.L. and Aarestrup, F.M. 1998.
Characterization of recently emerged multiple
antibiotic-resistant
Salmonella
enterica
serovar Typhimurium DT104 and other
multiresistant phage types from Danish pig
herds. The Veterinary Record, 143: 9597.

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015

2) Baggesen, D.L. and Wegener, H.C. 1994.

Phage types of Salmonella enterica ssp.
enterica serovar Typhimurium isolated from
production animals and humans in Denmark.
Acta Veterinaria Scandinavica, 35: 349354.
3) Bhunia, A. K. 2008. Foodborne microbial
pathogens: Mechanisms and pathogenesis.
United States of America: Springer Science +
Business Media, LLC.
4) Burley, S.K. 1994. DNA-binding motifs from
eukaryotic transcription factors. Curr Opin
Struct Biol, 4: 311.
5) Chia-Lung Li, Lien-I Hor, Zee-Fen Chang, LiChu Tsai Wei-Zen Yang, and Hanna S. Yuan.
2003. DNA binding and cleavage by the
periplasmic nuclease Vvn: a novel structure
with a known active site. EMBO J, 22:40144025.
6) Dominici, P., Tancini, B. and Voltatottorni, B
.C. 1985 .Chemical modification of pig kidney
3, 4-dihydroxyphenylalanine decarboxylase
with diethyl pyrocarbonate. J. Biol Chem.,
260: 10583-10589.
7) Dorman, C. J., Chatfield, S., Higgins, C.F.,
Hayward, C. and Dougan,
G.1989.
Characterization of porin and ompR mutants
of a virulent strain of Salmonella
typhimurium: ompR mutants are attenuated in
vivo. Infect. Immun. 57: 2136 2140.
8) Fang, F. C., Libby, S.J., Buchmeier, N.A.,
Loewen, P.C., Switala, J, J., Harwood, J. and
Guiney, D.G. 1992. The alternative sigma
factor katF (rpoS) regulates Salmonella
virulence. Proc. Natl. Acad. Sci. USA,
89:1197811982.
9) Fields, P. I., Swanson, R.V., Haidaris, C.G.
and Heffron, F. 1986. Mutants of Salmonella
typhimurium that cannot survive within the
macrophage are avirulent. Proc. Natl. Acad.
Sci. USA, 83:51895193.
10) Fried, M.1989. Measurement of protein-DNA
interaction parameter by Electrophoretic Gel
Mobility Shift. Electrophoresis, 10:366-376.
11) Galan, J. E. and Curtiss, R. III. 1989.
Virulence and vaccine potential of phoP

26

mutants of Salmonella typhimurium. Microb.

Pathog, 6:433443.
12) Gangathraprabhu, B and Ponmurugan, P.
2012. Gel Mobility assay of DNA- Pho P/Pho
Q Protein Interaction of MDR Salmonella
typhi isolated from Erode Slaughter House.
International
Journal
of
Biological
Technology, 3(1):45-49.
13) Garcia Vescovi, E., Soncini, F.C., and
Groisman, E.A. 1994. The role of the
PhoP/PhoQ regulon in Salmonella virulence.
Res. Microbiol., 145:473480.
14) Groisman, E. A., Chiao, E., Lipps, C.J. and
Heffron, F. 1989. Salmonella typhimurium
phoP virulence gene is a transcriptional
regulator. Proc. Natl. Acad. Sci. USA,
86:70777081.
15) Jones, S., Heyningen, P.V., Berman, H.M. and
Thornton,
J.M.
1999. ProteinDNA
interactions: a structural analysis. J MolBiol.,
287: 877-896.
16) Karim Roder and Michael Schweitzer. 2001.
Running-buffer composition influences DNA
protein and proteinprotein complexes
detected by electrophoretic
mobilityshift assay (EMSA). Biotechnol. Appl.
Biochem., 33:209214.
17) Miller, S. I., and Mekalanos, J.J. 1990.
Constitutive expression of the phoP regulon
attenuates Salmonella virulence and survival
within macrophages. J. Bacteriol., 172:2485
2490.
18) Molbak, K., Baggesen, D.L., Aarestrup, F.M.,
Ebbesen, J.M., Engberg, J., Frydendahl, K.,
Gerner-Smidt, P., Petersen, A.M. and
Wegener, H.C. 1999. An outbreak of
multidrug-resistant,
quinolone-resistant
Salmonella enterica serotype Typhimurium
DT104. The New England Journal of
Medicine, 341:14201425.
19) Olano, J., Soler, J., Busto, F. and De Arriaga,
D.1999 .Chemical modification of NADPisocitrate
dehydrogenase
from
Cephalosporium acremonium evidence of
essential histidine and lysine groups at the

2015 Published by JPS Scientific Publications Ltd. All rights reserved

Ponnusamy Ponmurugan /Life Science Archives (LSA), Volume 1, Issue 1, Page 13 to 27, 2015

27

active site. European Journal of Biochemistry,

261: 640-649.
20) Pabo,
C.O.
and
Nekludova,
L.
2000. Geometric analysis and comparison of
proteinDNA interfaces: why is there no
simple code for recognition? J Mol Biol., 301:
597624.
21) Pui, C. F., Wong, W. C., Chai, L. C., Nillian,
E., Ghazali, F. M., Cheah, Y. K., Nakaguchi,
Y., Nishibuchi, M. and Radu, S. 2011b.
Simultaneous detection of Salmonella spp.,
Salmonella
Typhi
and
Salmonella
typhimurium in sliced fruits using multiplex
PCR. Food Control, 22: 337-342.
22) Purification of nuclear DNA from excised
cotyledons of cucurbita. Bulg. J. Plant Physiol.
28(34):13.
23) Isobel A. Wadman, Hirotaka Osada, Gerald G.
Grutz, Alan D. Agulnick, Heiner Westphal
Alan Forster and Terence H.Rabbitts.1997.
The LIM-only protein Lmo2 is a bridging
molecule assembling an erythroid, DNAbinding complex which includes the TAL1,
E47, GATA-1 and Ldb1/NLI proteins. EMBO
J 16(11):31453157
24) Wolberger,
C.
1999. MultiproteinDNA
complexes in transcription regulation. Annu
Rev Biophys Biomol Struct, 28:2956.

2015 Published by JPS Scientific Publications Ltd. All rights reserved