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Antibiotic Susceptibility Pattern of Some Clinical Gram Positive Isolates

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 ايمان محمد جار الله مسير 21/01/2019 19:45:05
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ISSN: 0975-8542
Journal of Global Pharma Technology
Available Online at www.jgpt.co.in
Research Paper
©2009-2017, JGPT. All Rights Reserved 262
Antibiotic Susceptibility Pattern of Some Clinical Gram Positive
Isolates
Ehsan F. Hussein1*, Eman M. Jarallah2
1 University of Sumer, Iraq.
2 University of Babylon, Iraq.
*Corresponding Author: Ehsan F. Hussein (E mail: ehsan.algrani@yahoo.com)
Abstract
One hundred fifty samples have been collected through the period from July/2016 to December/2016.
Were for isolation and identification of pathogenic gram positive bacteria. The samples were involved
fifty samples from each skin, mouth and soil for isolate Staphylococcus aureus, Streptococcus pyogenes
and Bacillus spp., respectively. Isolates were identified according to microscopic features and biochemical
tests .Antibiotics susceptibility pattern were examined by disc diffusion method. The antibiotics
explained different action modes and some of the have significant differences when comparisons
together. The Trimethoprim antibiotic has high activity against Staph. aureus, and Strep. pyogenes while
the Carbnicillin, Rifaxim in and Trimethoprim antibiotics have high activity against Bacillus spp. These
antibiotics were Amoxicillin, Ampicillin, Trimethoprim, Aztreonam, Phamethoxazole, Carbnicillin,
Novobiocin, Erythromycin, Rifaximin, Penicillin-G, Oxolinic Acid, Bacitracin and Clindamycin.
Trimethoprim antibiotic has high activity when comparison the activity of these antibiotics together.
Keywords: Pathogenic bacteria, Trimethoprim, Antibiotics, Resistant.
Introduction
The resistant in bacteria is occurring
worldwide endangering the efficacy of
antibiotics which have transformed medicines
and save lives of millions [1]. Many reports
showed that the after use of antibiotics in
treated the first patient, the infections of
bacteria become a threat again [2].
The causes of antibiotics resistance have been
attributed to the misuse of these medications
and overuse as well as a loss of development
of new drug by the pharmaceutical industry
caused by reduced challenging regulatory
requirements and economic incentives [3].
Emergency of bacterial resistant threat the
extraordinary health benefit that has been
achieved with antibiotics treatment [4].
Staphylococcus is heterogeneous bacterial
group and consists of thirty species but the
Staphylococcus aurous is the most clinically
important species [5]. This in the family of
Staphylococcaceae, an important opportunistic
pathogen and can causes infections for both
human and animals [6]. Staph. aureus is gram
positive with diameter 0.8-1 ?m, non-motile,
non-speculated and non-capsulated [7].
Staphylococcus aurous was considered has
over the past studies as one of the most
important pathogens for human and leading to
causes of community acquired infections and
hospital [8]. Infections of staphylococcal
associated with wide spectrum of diseases in
humans [9].
This bacterium can colonize of the skin and
mucous membrane and from which it have
ability to invade other organs in body [10], and
considered as an important causative agent
for skin and soft tissue infections [10].In
despite of, Staphylococcus infections can easily
treated with antibiotics, but in recent years,
this bacterium developed resist against most
commonly used and effective antibiotics [11-
12].
Now, the methicillin resistant Staph. aurous
(MRSA) has become prevalence problem in
hospitals and wide range of acquired
infections [13]. Furthermore, the multi drug
Ehsan F. Hussein et. al.: Journal of Global Pharma Technology. 2017; 10(9):262-274
©2009-2017, JGPT. All Rights Reserved 263
methicillin resistant Staph. aureus with very
limited treatment choice also increased [14].
Streptococcus pyogenes the major causes
pathogen and associated with acute
respiratory tract infections [15]. This
bacterium can cause several types of
infections such as pharyngitis and pyoderma
[16]. This microorganism have been
distinguished as one of infectious agents that
leading to puerperal fever in human and also
contributing in the skin, ear, vagina and
throat infections[17]. Invasive infections like
streptococcal toxic shock syndrome and sepsis
began to receive attention from
epidemiologists and recently reported as
higher incidence rates and even epidemics
these bacteria [18].
The number of such infections associated
with Streptococcus pyogenes especially in
developed countries caused May by loses of
diagnostic methods and therapeutic problem
[19]. Increasing the resistance of antibiotics
among Streptococcus pyogenes have been
observed during the last decade in worldwide
[20]. The increase in the rate of antibiotic
resistant among clinical isolates of
Streptococcus pyogenes, this contributes in
the needs for continuous studies for known
the patterns of antimicrobial resistance [21].
Species of Bacillus bacteria are spore
forming, gram-positive and chemo
heterotrophic bacilli which are usually
aerobic or facultative anaerobic [22]. The
members of these bacteria are generally
found in soils and have abilities for grow in
every environment as well as have capability
to formation resistant spores and metabolite
products [23].
Bacillus spp. is a gram positive rods bacteria
usually found in soil [24]. These bacteria
responsible for No socomial infections [25].
Also, the spores produced by Bacillus are
frequently causes foods contamination which
resulting in diseases like diarrhea and
vomiting[26]. Bacillus spp. have ability to
resistant several types of antibiotic [27].
However, there are limited information about
antibiotics resistance by these bacteria [28].
Materials and Methods
Isolation of Pathogenic Positive Bacteria
One hundred fifty samples have been collected
were for isolation of pathogenic gram positive
bacteria. The samples were involve fifty skin
samples for is oblate Staphylococcus aureus,
fifty mouth samples for isolate Streptococcus
pyogenes and fifty soil samples for Bacillus
spp.
Figure1: Number and percent of pathogenic gram positive isolated bacteria
Figure (1) the number and percent of Staph.
aurous isolated from Skin, Strep. Pyogenes
isolated from Mouth and Bacillus spp.
isolated from Soil were (20; 33%), (16; 27%)
and (24; 40%) respectively.
Identification of bacterial Isolates
The isolates of Staph aureus were
characterized and identified according to [29].
This done by use of microscope and
biochemical tests such as gram staining,
coagulates test and catalase test [9].
The isolates of Strep Pyogenes were identified
and determined according to [30]. This carries
out by use of microscope and biochemical tests
such as gram staining, catalase test and blood
agar hemolytic [31]. The isolates of Bacillus
spp were isolated by use of serial diluted of
soil samples in sterile distilled water and
plated on nutrients agar media [32]. As well as
descriptive and identified according to [33].
This worked by use of microscope and
biochemical tests such as gram staining,
catalase test, oxidase test and indol test [34].
Ehsan F. Hussein et. al.: Journal of Global Pharma Technology. 2017; 10(9):262-274
©2009-2017, JGPT. All Rights Reserved 264
Antibiotic Susceptibility Test
Antibiotic susceptibility tests for pathogenic
gram positive bacteria were carried out by
technique of disc diffusion method on Muller
Hinton agar plates [35].
Statistical Analysis
The results were analyzed by the Statistical
Package Social Sciences (SPSS) version 20 for
determine of Mean, Medium, Standard
deviation and Standard error of mean, in
addition the significance between antibiotics
calculated according to One Way ANOVA by
descriptive excluded cases analysis by
analysis with LSD at (95%) confidence and
significant level at (P=0.05).
Figure 2: Inhibition zones formed by antibiotic types
Results
One hundred fifty samples have been
collected were for isolation of pathogenic
gram positive bacteria. This carries out
through the period from July/2016 to
December/2016. The samples were involve
fifty skin samples of isolate Staphylococcus
aureus, fifty mouth samples of isolate
Streptococcus pyogenes and fifty soil samples
of Bacillus spp. these isolated were
characterized and identified by use of several
type of biochemical tests as well as the
antibiotics sensitivity and resistant were
doing by use 13 types of antibiotics. These
antibiotics have different modes of action as
well as showed the Trimethoprim antibiotic
has high activity against both Staphs. aureus
and Strep. pyogenes while the Carbnicillin
antibiotic has high activity against Bacillus
spp. Also explained the Trimethoprim
antibiotic has high activity when comparison
the activity of these antibiotics together. Also
the antibiotics have significant differences
and their activity against the bacteria. Figure
(2) the inhibition zones that formed by
different types of antibiotics on Muller
Hinton Agar after (24 h) against
experimental pathogenic gram positive
bacteria when cultured with antibiotics
together on this medium.
Table1: Replicates of inhibition zones formed by antibiotic types against Staph. aurous
Inhibition zones replicates measured by (mm)
CC 5
?g
B 10
?g
OA 2
?g
P 10
?g
R 40
?g
E 15
?g
NV 30
?g
PY 100
?g
SMZ 25
?g
ATM 30
?g
TMP 10
?g
AM 10
?g
AX 15
?g
Replicates/*
1 6 7 16 9 6 6 0 7 4 8 6 5 5
2 7 8 6 12 0 2 7 0 9 2 6 3 8
3 8 10 10 9 0 3 11 0 0 1 0 12 4
4 20 10 25 6 25 4 4 0 4 8 11 3 11
5 25 11 25 4 6 2 6 0 5 0 6 4 3
6 16 13 20 13 11 7 9 5 7 6 9 6 2
7 6 5 8 14 22 0 20 0 5 12 18 0 11
8 32 26 32 20 0 14 0 10 6 0 0 6 6
9 0 18 11 24 18 4 10 7 6 8 10 9 0
10 10 8 29 4 4 10 8 3 9 6 8 2 8
11 18 15 22 13 13 9 5 8 18 2 11 8 9
12 12 8 19 16 20 18 6 0 7 16 0 7 7
13 30 29 30 6 6 0 20 16 16 0 22 4 13
14 15 10 14 12 16 9 10 6 12 4 18 2 15
15 18 13 25 2 10 9 14 12 8 10 6 14 5
Mean 14.866 12.734 19.467 10.933 10.466 6.467 8.666 4.933 7.7334 5.533 8.734 5.666 7.133
Median 15.000 10.000 20.000 12.000 10.000 6.000 8.000 5.000 7.000 6.000 8.000 5.000 7.000
9.226 6.861 8.339 6.158 8.279 5.139 5.961 5.119 4.667 4.853 6.649 3.848 4.189
Std.
Deviation
2.382 1.771 2.153 1.591 2.137 1.326 1.538 1.321 1.205 1.253 1.716 0.993 1.081
Std. Error of
Mean
Ehsan F. Hussein et. al.: Journal of Global Pharma Technology. 2017; 10(9):262-274
©2009-2017, JGPT. All Rights Reserved 265
Table (1) the Mean, Median, Standard
deviation and Standard error for replicates of
inhibition zones were measured by (mm) of
each antibiotics against Staph. aurous
bacteria and found the Trimethoprim
antibiotic has high activity (19.467 mm)
while the (Erythromycin) antibiotic has low
activity (4.933mm) against this bacterium by
measured of mean.
? *AX 15=Amoxicillin15 *E 15= Erythromycin15
? *AM 10= Ampicillin10 *R 40=Rifaximin40
? *TMP 10= Trimethoprim10 *P 10=Penicillin-G10
? *ATM 30= Aztreonam30 *OA 2= Oxolinic Acid2
? *SMZ 25= Phamethoxazole25 *B 10=Bacitracin10
? *PY 100= Carbnicillin100 *CC 5= Clindamycin5
? *NV 30=Novobiocin3
Table2: LSD system for study of significant differences between of antibiotic types against Staph. aurous
Significantly results according to LSD system at P-Value= 0.05
Y side / X side
(antibiotics)
AX15
?g
AM10
?g
TMP10
?g
ATM30
?g
SMZ25
?g
PY100
?g
NV30
?g
E 15
?g
R 40
?g
P 10
?g
OA 2
?g
B 10
?g
CC 5
?g
AX15
?g
Sig. 0.356 0.048* 0.090 0.058 0.001* 0.008* 0.001* 0.002* 0.001* 0.009* 0.001* 0.001*
M.D. ** -2.133 4.601 -3.933 -4.401 -8.401 -6.201 -9.933 -7.133 -9.333 -6.133 -9.201 -7.733
AM10
?g
Sig. 0.356 0.004* 0.436 0.327 0.007* 0.079 0.001* 0.031* 0.002* 0.048* 0.003* 0.016*
M.D. ** 2.133 6.733 -1.801 -2.266 -6.266 -4.066 -7.801 -5.001 -7.201 -4.001 -7.066 -5.601
TMP10
?g
Sig. 0.048* 0.004* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001*
M.D. ** -4.601 -6.733 -8.533 -9.001 -13.001 -10.801 -14.533 -11.733 -13.933 -10.733 -13.801 -12.333
ATM30
?g
Sig. 0.090 0.436 0.001* 0.840 0.054 0.327 0.001* 0.167 0.001* 0.341 0.001* 0.101
M.D. ** 3.933 1.801 8.533 -0.466 -4.466 -2.266 -6.001 -3.201 -5.401 -2.201 -5.266 -3.801
SMZ25
?g
Sig. 0.058 0.327 0.001* 0.840 0.084 0.436 0.001* 0.237 0.034* 0.453 0.039* 0.150
M.D. ** 4.401 2.266 9.001 0.466 -4.001 -1.801 -5.533 -2.733 -4.933 -1.733 -4.801 -3.333
PY100
?g
Sig. 0.001* 0.007* 0.001* 0.054 0.084 0.341 0.507 0.583 0.686 0.327 0.729 0.773
M.D. ** 8.401 6.266 13.001 4.466 4.001 2.201 -1.533 1.266 0.933 2.266 -0.801 0.666
NV30
?g
Sig. 0.008* 0.079 0.001* 0.327 0.436 0.341 0.107 0.686 0.176 0.977 0.195 0.507
M.D. ** 6.201 4.066 10.801 2.266 1.801 -2.201 -3.733 -0.933 -3.133 0.066 -3.001 -1.533
E15
?g
Sig. 0.001* 0.001* 0.001* 0.001* 0.001* 0.507 0.107 0.226 0.795 0.101 0.751 0.341
M.D. ** 9.933 7.801 14.533 6.001 5.533 1.533 3.733 2.801 0.601 3.801 0.733 2.201
R40
?g
Sig. 0.002* 0.031* 0.001* 0.167 0.237 0.583 0.686 0.226 0.341 0.665 0.371 0.795
M.D. ** 7.133 5.001 11.733 3.201 2.733 -1.266 0.933 -2.801 -2.201 1.001 -2.066 -0.601
P10
?g
Sig. 0.001* 0.002* 0.001* 0.001* 0.034* 0.686 0.176 0.795 0.341 0.167 0.954 0.489
M.D. ** 9.333 7.201 13.933 5.401 4.933 0.933 3.133 -0.601 2.201 3.201 0.133 1.601
OA2
?g
Sig. 0.009* 0.048* 0.001* 0.341 0.453 0.327 0.977 0.101 0.665 0.167 0.185 0.489
M.D. ** 6.133 4.001 10.733 2.201 1.733 -2.266 -0.066 -3.801 -1.001 -3.201 -3.066 -1.601
B10
?g
Sig. 0.001* 0.003* 0.001* 0.001* 0.039* 0.729 0.195 0.751 0.371 0.954 0.185 0.525
M.D. ** 9.201 7.066 13.801 5.266 4.801 0.801 3.001 -0.733 2.066 -0.133 3.066 1.466
CC5
?g
Sig. 0.001* 0.016* 0.001* 0.101 0.150 0.773 0.507 0.341 0.795 0.489 0.489 0.525
M.D. ** 7.733 5.601 12.333 3.801 3.333 -0.666 1.533 -2.201 0.601 -1.601 1.601 -1.466
*Significant differences of mean at the p-valve equal to 0.05 level
**Differences of Mean
Table (2) the significant differences between
types of different antibiotics when study the
activities of these antibiotics against Staph.
aurous by statistical analysis by used of LSD
system.Also explained the significant
differences that found between used
antibiotics and this showed through the star
that present at p-valve equal to 0.05 levels.Not
all antibiotics that studied have
significant differences through comparison
together, some of these antibiotics have
significant differences and this showed
through the star that found at p-valve equal to
0.05 levels. Comparisons involved two types.
First, involve the (X) side antibiotics with the
(Y) side antibiotics and second involved
comparison the (Y) side antibiotics with the
(X) side antibiotics.
Figure 3: Action modes of antibiotic types against Staph. aurous
Ehsan F. Hussein et. al.: Journal of Global Pharma Technology. 2017; 10(9):262-274
©2009-2017, JGPT. All Rights Reserved 266
Figure (3) the types of antibiotics give
different action modes, this mean each
antibiotic explained inhibition zones with
different size against same bacteria at fifteen
replicates, as well as (AX 15 & TMP 10)
antibiotics have high inhibition zones were
(32 mm).
Figure4: Percent of antibiotic types against Staph. aureus
Figure (4) action percentages of different
types of antibiotic; also showed (TMP 10)
antibiotics has high percent equal to 16% as
well as the (E 15) antibiotic has low percent
equal to 4% from all activities of antibiotics
against Staph. aureus.
Table3: Replicates of inhibition zones formed by antibiotic types against Strep. pyogenes
Inhibition zones replicates measured by (mm)
CC 5
?g
B 10
?g
OA 2
?g
P 10
?g
R 40
?g
E 15
?g
NV 30
?g
PY 100
?g
SMZ 25
?g
ATM 30
?g
TMP 10
?g
AM 10
?g
AX 15
?g
Replicates/*
1 0 7 16 10 10 3 16 4 6 0 0 0 3
2 6 4 18 12 8 4 15 0 7 5 3 10 0
3 12 0 10 7 12 5 18 10 10 6 0 0 5
4 5 8 25 10 13 7 13 3 6 5 3 2 8
5 0 9 25 12 7 2 14 0 9 8 0 0 9
6 5 3 24 22 16 9 17 3 6 4 3 3 6
7 4 9 23 16 11 4 19 2 11 9 2 8 4
8 10 2 20 12 10 5 12 10 5 7 7 2 5
9 3 10 19 9 8 0 15 6 4 4 2 5 4
10 9 2 20 14 12 7 13 2 3 2 1 3 2
11 5 7 24 15 16 10 20 8 13 10 5 7 3
12 11 4 28 12 18 5 18 4 0 2 4 5 6
13 8 8 24 11 20 7 23 2 8 5 3 3 0
14 4 0 20 18 14 3 15 3 9 3 6 4 4
15 7 4 21 20 12 2 19 4 3 6 2 0 12
Mean 5.934 5.133 21.134 13.333 12.467 4.866 16.467 4.066 6.667 5.066 2.734 3.466 4.733
Median 5.000 4.000 21.000 12.000 12.000 5.000 16.000 3.000 6.000 5.000 3.000 3.000 4.000
Std. Deviation 3.614 3.356 4.421 4.186 3.777 2.748 3.044 3.151 3.436 2.737 2.121 3.217 3.217
Std. Error of 0.334 0.866 1.141 1.081 0.975 0.709 0.785 0.813 0.887 0.706 0.546 0.798 0.831
Mean
Table (3) the Mean, Median, Standard
deviation and Standard error for replicates of
inhibition zones were measured by (mm) for
each antibiotics against Strep. pyogenes
bacteria. And found the Trimethoprim
antibiotic has high activity (21.134 mm) while
0.00%
5.00%
10.00%
15.00%
20.00%
CC 5?g
B 10?g
OA 2?g
P 10 ?g
R 40?g
E 15?g
NV 30?g PY 100?g
SMZ 25?g
ATM 30?g
TMP 10?g
AM 10?g
Staph. aureus AX 15?g
Ehsan F. Hussein et. al.: Journal of Global Pharma Technology. 2017; 10(9):262-274
©2009-2017, JGPT. All Rights Reserved 267
the (Oxolinic Acid) antibiotic has low activity
(2.734mm) against this bacterium by
measured of mean.
? *AX 15=Amoxicillin15 *E 15= Erythromycin15
? *AM 10= Ampicillin10 *R 40=Rifaximin40
? *TMP 10= Trimethoprim10 *P 10=Penicillin-G10
? *ATM 30= Aztreonam30 *OA 2= Oxolinic Acid2
? *SMZ 25= Phamethoxazole25 *B 10=Bacitracin10
? *PY 100= Carbnicillin100 *CC 5= Clindamycin5
? *NV 30=Novobiocin3
Table 4: LSD system for study of significant differences between of antibiotic types against Strep pyogenes
Significantly results according to LSD system at P-Value= 0.05
Y side / X side
(antibiotics)
AX15
?g
AM10
?g
TMP10
?g
ATM30
?g
SMZ25
?g
PY100
?g
NV30
?g
E 15
?g
R 40
?g
P 10
?g
OA 2
?g
B 10
?g
CC 5
?g
AX15
?g
Sig. 0.514 0.001* 0.001* 0.001* 0.385 0.001* 0.129 0.550 0.480 0.011* 0.045* 0.328
M.D. ** -0.801 15.201 7.4001 6.533 -1.066 10.533 -1.866 0.733 -0.866 -3.201 -2.466 -1.201
AM10
?g
Sig. 0.514 0.001* 0.001* 0.001* 0.828 0.001* 0.385 0.212 0.957 0.051 0.175 0.744
M.D. ** 0.801 16.001 8.201 7.333 -0.266 11.333 -1.066 1.533 -0.066 -2.401 -1.666 -0.401
TMP10
?g
Sig. 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001*
M.D. ** -15.201 -16.001 -7.801 -8.666 -16.266 -4.666 -17.066 -14.466 -16.066 -18.401 -17.666 -16.401
ATM30
?g
Sig. 0.001* 0.001* 0.001* 0.480 0.001* 0.011* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001*
M.D. ** -7.401 -8.201 7.801 -0.866 -8.466 3.133 -9.266 -6.666 -8.266 -10.601 -9.866 -8.601
SMZ25
?g
Sig. 0.001* 0.001* 0.001* 0.480 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001*
M.D. ** -6.533 -7.333 8.666 0.866 -7.601 4.001 -8.401 -5.801 -7.401 -9.733 -9.001 -7.733
PY100
?g
Sig. 0.385 0.828 0.001* 0.001* 0.001* 0.001* 0.514 0.143 0.870 0.083 0.254 0.913
M.D. ** 1.066 0.266 16.266 8.466 7.601 11.601 -0.801 1.801 0.201 -2.133 -1.401 -0.133
NV30
?g
Sig. 0.001* 0.001* 0.001* 0.011* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001* 0.001*
M.D. ** -10.533 -11.333 4.666 -3.133 -4.001 -11.601 -12.401 -9.801 -11.401 -13.733 -13.001 -11.733
E15
?g
Sig. 0.129 0.385 0.001* 0.001* 0.001* 0.514 0.001* 0.035* 0.415 0.278 0.625 0.587
M.D. ** 1.866 1.066 17.066 9.266 8.401 0.801 12.401 2.601 1.001 -1.333 -0.600 0.666
R40
?g
Sig. 0.550 0.212 0.001* 0.001* 0.001* 0.143 0.001* 0.035* 0.193 0.002* 0.011* 0.116
M.D. ** -0.733 -1.533 14.466 6.666 5.801 -1.801 9.801 -2.601 -1.601 -3.933 -3.201 -1.933
P10
?g
Sig. 0.480 0.957 0.001* 0.001* 0.001* 0.870 0.001* 0.415 0.193 0.058 0.193 0.786
M.D. ** 0.866 0.066 16.066 8.266 7.401 -0.201 11.401 -1.001 1.601 -2.333 -1.601 -0.333
OA2
?g
Sig. 0.011* 0.051 0.001* 0.001* 0.001* 0.083 0.001* 0.278 0.002* 0.058 0.550 0.104
M.D. ** 3.201 2.401 18.401 10.601 9.733 2.133 13.733 1.333 3.933 2.333 -0.733 -0.551
B10
?g
Sig. 0.045* 0.175 0.001* 0.001* 0.001* 0.254 0.001* 0.625 0.011* 0.193 0.550 0.302
M.D. ** 2.466 1.666 17.666 9.866 9.001 1.401 13.001 0.600 3.201 1.601 -0.733 1.266
CC5
?g
Sig. 0.328 0.744 0.001* 0.001* 0.001* 0.913 0.001* 0.587 0.116 0.786 0.104 0.302
M.D. ** 1.201 0.401 16.401 8.601 7.733 0.133 11.733 -0.666 1.933 0.333 0.551 -1.266
*Significant differences of mean at the p-valve equal to 0.05 level
**Differences of Mean
Table (4) the significant differences between
types of different antibiotics when study the
activities of these antibiotics against Strep.
pyogenes by statistical analysis by used of
LSD system.
Also explained the significant differences that
found between used antibiotics and this
showed through the star that present at pvalve
equal to 0.05 levels. Not all antibiotics
that studied have significant differences
through comparison together, some of these
antibiotics have significant differences and
this showed through the star that found at pvalve
equal to 0.05 levels.
Comparisons involved two types. First,
involve the (X) side antibiotics with the (Y)
side antibiotics and second involved
comparison the (Y) side antibiotics with the
(X) side antibiotics.
Figure 5: Action modes of antibiotic types against Strep. Pyogenes
Ehsan F. Hussein et. al.: Journal of Global Pharma Technology. 2017; 10(9):262-274
©2009-2017, JGPT. All Rights Reserved 268
Figure (5) the types of antibiotics give
differentiation modes, this mean each
antibiotic explained inhibition zones with
different size against same bacteria at fifteen
replicates, as well as (TMP 10) antibiotics has
high inhibition zone was (28 mm).
Figure 6: Percent of antibiotic types against Strep. Pyogenes
Figure (6) action percentages of different types
of antibiotic; also showed (TMP 10) antibiotics
has high percent equal to 20% as well as the
(OA 2) antibiotic has low percent equal to 3%
from all activities of antibiotics against Strep.
pyogenes.
Table 5: Replicates of inhibition zones formed by antibiotic types against Bacillus spp
Inhibition zones replicates measured by (mm)
CC 5
?g
B 10
?g
OA 2
?g
P 10
?g
R 40
?g
E 15
?g
NV 30
?g
PY 100
?g
SMZ 25
?g
ATM 30
?g
TMP 10
?g
AM 10
?g
AX 15
?g
Replicates/*
1 4 7 7 3 8 12 5 5 10 8 4 3 0
2 12 3 17 0 3 13 4 9 8 8 13 5 3
3 6 7 8 6 9 7 3 8 6 13 6 4 0
4 13 6 2 0 4 10 3 5 12 5 9 5 2
5 5 5 5 11 3 8 8 15 9 13 10 0 4
6 8 0 13 5 6 12 0 7 10 8 8 6 5
7 16 6 16 4 4 14 4 6 12 6 16 4 3
8 10 7 15 7 6 19 11 10 16 6 10 8 7
9 6 6 8 6 8 9 5 6 14 15 6 9 2
10 6 8 7 9 5 18 4 7 9 6 7 3 8
11 3 9 11 5 5 13 6 9 13 7 10 8 6
12 8 6 8 4 12 6 6 5 11 11 8 5 5
13 12 10 15 7 5 20 6 0 15 6 15 6 5
14 9 13 11 0 2 10 8 8 7 9 9 0 8
15 0 5 9 6 9 16 4 7 18 4 8 8 9
Mean 7.866 6.533 10.134 4.866 5.933 12.467 5.133 7.134 11.333 8.334 9.266 4.933 4.466
Median 8.000 6.000 9.000 5.000 5.000 12.000 5.000 7.000 11.000 8.000 9.000 5.000 5.000
Std. Deviation 4.241 2.972 4.356 3.204 2.763 4.323 2.587 3.226 3.415 3.265 3.305 3.326 2.825
1.064 0.767 1.124 0.827 0.713 1.116 0.668 0.883 0.881 0.843 0.858 0.853 0.792
Std. Error of
Mean
Table (5) the Mean, Median, Standard
deviation and Standard error for replicates of
inhibition zones were measured by (mm) for
each antibiotics against Bacillus spp.
bacteria. And found the Carbnicillin has high
activity was (12.467mm) while the
Clindamycin antibiotics have low activity
0.00%
5.00%
10.00%
15.00%
20.00%
25.00%
CC 5?g
B 10?g
OA 2?g
P 10 ?g
R 40?g
E 15?g
NV 30?g PY 100?g
SMZ 25?g
ATM 30?g
TMP 10?g
AM 10?g
Strep. pyogenes AX 15?g
Ehsan F. Hussein et. al.: Journal of Global Pharma Technology. 2017; 10(9):262-274
©2009-2017, JGPT. All Rights Reserved 269
(4.466mm) against this bacterium by
measured of mean.
? *AX 15=Amoxicillin15 *E 15= Erythromycin15
? *AM 10= Ampicillin10 *R 40=Rifaximin40
? *TMP 10= Trimethoprim10 *P 10=Penicillin-G10
? *ATM 30= Aztreonam30 *OA 2= Oxolinic Acid2
? *SMZ 25= Phamethoxazole25 *B 10=Bacitracin10
? *PY 100= Carbnicillin100 *CC 5= Clindamycin5
? *NV 30=Novobiocin3
Table 6: LSD system for study of significant differences between of antibiotic types against Bacillus spp
Significantly results according to LSDsystem at P-Value= 0.05
Y side / X side
(antibiotics)
AX15
?g
AM10
?g
TMP10
?g
ATM30
?g
SMZ25
?g
PY100
?g
NV30
?g
E 15
?g
R 40
?g
P 10
?g
OA 2
?g
B 10
?g
CC 5
?g
AX15
?g
Sig. 0.287 0.071 0.017* 0.123 0.001* 0.031* 0.557 0.006* 0.709 0.263 0.021* 0.007*
M.D. ** -1.333 2.266 -3.001 -1.933 4.601 -2.733 -0.733 3.466 0.466 1.400 -2.933 -3.401
AM10
?g
Sig. 0.287 0.004* 0.183 0.631 0.001* 0.263 0.631 0.001* 0.151 0.030* 0.201 0.099
M.D. ** 1.333 3.601 -1.666 0.601 5.933 -1.401 0.601 4.801 1.801 2.733 -1.601 -2.066
TMP10
?g
Sig. 0.071 0.004* 0.001* 0.001* 0.063 0.001* 0.017* 0.337 0.151 0.488 0.001* 0.001*
M.D. ** -2.266 -3.601 -5.266 -4.201 2.333 -5.001 -3.001 1.201 -1.801 -0.866 -5.201 -5.666
ATM30
?g
Sig. 0.017* 0.183 0.001* 0.394 0.001* 0.831 0.071 0.001* 0.006* 0.001* 0.957 0.749
M.D. ** 3.001 1.666 5.266 1.066 7.601 0.266 2.266 6.466 3.466 4.401 0.066 -0.401
SMZ25
?g
Sig. 0.123 0.631 0.001* 0.394 0.001* 0.522 0.337 0.001* 0.056 0.008* 0.424 0.241
M.D. ** 1.933 0.601 4.201 -1.066 6.533 -0.801 1.201 5.401 2.401 3.333 -1.001 -1.466
PY100
?g
Sig. 0.001* 0.001* 0.063 0.001* 0.001* 0.001* 0.001* 0.365* 0.001* 0.001* 0.001* 0.001*
M.D. ** -4.601 -5.933 -2.333 -7.601 -6.533 7.333 5.333 -1.133 -4.133 -3.201 -7.533 -8.001
NV30
?g
Sig. 0.031* 0.263 0.001* 0.831 0.522 0.001* 0.111 0.001* 0.011* 0.001* 0.873 0.594
M.D. ** 2.733 1.401 5.001 -0.266 0.801 7.333 2.001 6.201 3.201 4.133 -0.201 -0.666
E15
?g
Sig. 0.557 0.631 0.017* 0.071 0.337 0.001* 0.111 0.001* 0.337 0.089 0.080 0.034*
M.D. ** 0.733 -0.601 3.001 -2.266 1.201 -5.333 -2.001 4.201 1.201 2.133 -2.201 -2.666
R40
?g
Sig. 0.006* 0.001* 0.337 0.001* 0.001* 0.365* 0.001* 0.001* 0.017* 0.099 0.001* 0.001*
M.D. ** -3.466 -4.801 -1.201 -6.466 -5.401 1.133 -6.201 -4.201 -3.001 -2.066 -6.401 -6.866
P10
?g
Sig. 0.709 0.151 0.151 0.006* 0.056 0.001* 0.011* 0.337 0.017* 0.455 0.007* 0.002*
M.D. ** -0.466 -1.801 1.801 -3.466 -2.401 4.133 -3.201 -1.201 3.001 0.933 -3.401 -3.866
OA2
?g
Sig. 0.263 0.030* 0.488 0.001* 0.008* 0.001* 0.001* 0.089 0.099 0.455 0.001* 0.001*
M.D. ** -1.400 -2.733 0.866 -4.401 -3.333 3.201 -4.133 -2.133 2.066 -0.933 -4.333 -4.801
B10
?g
Sig. 0.021* 0.201 0.001* 0.957 0.424 0.001* 0.873 0.080 0.001* 0.007* 0.001* 0.709
M.D. ** 2.933 1.601 5.201 -0.066 1.001 7.533 0.201 2.201 6.401 3.401 4.333 -0.466
CC5
?g
Sig. 0.007* 0.099 0.001* 0.749 0.241 0.001* 0.594 0.034* 0.001* 0.002* 0.001* 0.709
M.D. ** 3.401 2.066 5.666 0.401 1.466 8.001 0.666 2.666 6.866 3.866 4.801 0.466
*Significant differences of mean at the p-valve equal to 0.05 level
**Differences of Mean
Table (6) the significant differences between
types of different antibiotics when study the
activities of these antibiotics against Bacillus
spp. By statistical analysis by used of LSD
system.
Also explained the significant differences that
found between used antibiotics and this
showed through the star that present at pvalve
equal to 0.05 levels.
Not all antibiotics that studied have
significant differences through comparison
together, some of these antibiotics have
significant differences and this showed
through the star that found at p-valve equal
to 0.05 levels.
Comparisons involved two types. First,
involve the (X) side antibiotics with the (Y)
side antibiotics and second involved
comparison the (Y) side antibiotics with the
(X) side antibiotics.
Figure7: Action modes of antibiotic types against Bacillus spp
Ehsan F. Hussein et. al.: Journal of Global Pharma Technology. 2017; 10(9):262-274
©2009-2017, JGPT. All Rights Reserved 270
Figure (7) the types of antibiotics give
differentiation modes, this mean each
antibiotic explained inhibition zones with
different size against same bacteria at fifteen
replicates, as well as (PY 100) antibiotics has
high inhibition zone was (20 mm).
Figure 8: Percent of antibiotic types against Bacillus spp
Figure (8) action percentages of different
types of antibiotic; also showed (PY 100)
antibiotics has high percent equal to 13%
as well as the (CC 5) antibiotic has low
percent equal to 5% from all activities of
antibiotics against Bacillus spp.
Figure 9: Comparisons between all antibiotics means against pathogenic positive bacteria
Figure (9) the Trimethoprim antibiotic has
high activity when comparison the activity of
the all antibiotic types against pathogenic
gram positive bacteria.
Discussion
In this study the trimethoprim antibiotic has
high activity against Staphylococcus aurous
(Table: 1 and Figure: 4). These were agreed
with the Kaka et al who found this antibiotic
has bactericidal activity against these
bacteria [36]. While dis agreed with Tiwar et
al who presented some isolate of this bacteria
were resistant to trimethoprim [37]. The
current study explained the erythromycin
antibiotic has low activity against Staph.
aureus (Table: 1 and Figure: 4). These were
agreed with Brown who found the isolate of
this bacteria were resistant to this antibiotic
[38]. Staphylococcus aureus is the major
causes of acquired infections in hospital and
demonstrated this pathogen in community
acquired infection [39]. These bacteria now
have capacity to resistant most antibiotic
types [40].
In the present study the trimethoprim
antibiotic has high activity against
Streptococcus Pyogenes (Table:3 and
Figure:6). These results were agreed with
Bowen et al who found these bacteria were
Susceptible to this antibiotic [41]. But dis
agreed with study of Bergmann who found
these bacteria were resistant to this
antibiotic [42]. And with Dangwetngam et al
who found this antibiotic has no activity but
against Streptococcus agalactiae [43]. The
0.00%
5.00%
10.00%
15.00%
CC 5?g
B 10?g
OA 2?g
P 10 ?g
R 40?g
E 15?g
NV 30?g PY 100?g
SMZ 25?g
ATM 30?g
TMP 10?g
AM 10?g
Bacillus spp. AX 15?g
Ehsan F. Hussein et. al.: Journal of Global Pharma Technology. 2017; 10(9):262-274
©2009-2017, JGPT. All Rights Reserved 271
current study showed the Oxolinic acid
antibiotic has low activity against Strep.
pyogenes. These were agreed with Matsuoka
and Wada who found this antibiotic has low
activity but against Streptococcus iniae [44].
Also agreed with Dangwetngamet al but
against the Streptococcus agalactiae [43].
Streptococcus pyogenes can causes infection
and has ability for large number of diseases
[16]. Also this bacteria can resistant wide
range of antibiotic types [15].
In the current study the Carbnicillin
antibiotic has high activity against Bacillus
spp. (Table:5 andFigure:8). These results
were agreed with Naik et al who found the
isolates of B. cereus were sensitive to this
antibiotic, but dis agreed with Naik et al who
found the isolate of B. subtilis were resistant
to this antibiotic[45]. In the current study
showed the Clindamycin antibiotic has low
activity against Bacillus spp. This was agreed
with Kervick et al who explained the isolates
of Bacillus spp. were resistant to
Clindamycin [46]. However, Bacillus spp. are
the widely distributed in soil and have ability
to resistant of essential antibiotics [47]. In
addition, these bacteria can causes and
associated with several types of infections
[48].
Figures (2, 3, 5, and 7) showed the antibiotic
types in current study have different action
modes. These differences may cause by types
of antibiotics that used and types of bacteria
under study as well as type of antibiotics
targets, however, some type of antibiotics act
by destroyed the nucleic acids DNA and/or
RNA, inactive protein synthesis, cell wall
breaking, cell wall destroyed and stopped of
essential path ways [49]. Antibiotics can be
classified to whether they induce death of cell
called bactericidal antibiotics or inhibit
growth of cell called bacteriostatic antibiotics
[50]. Also, this depended on mechanisms of
antimicrobial resistance such as, multi
antibiotic efflux pumps, antibiotic modified or
breakdown enzymes and change or invisible
of antibiotic targeting [51].
Figure (9) explained the trimethoprim
antibiotic has high activity when comparison
the activity of all antibiotic types together.
Trimethoprim has activity against wide
range of gram positive aerobic bacteria and
comparable in efficacy with ampicillin,
oxolinic acid and cephalexin [52]. However,
trimethoprim has inhibitory activity against
wide range of infections such as bacterial of
typhoid fever, bacterial pneumonia and
chronic bronchitis, as well as being
investigated as definitive therapy against
most infections [53].
Conclusions
The Trimethoprim antibiotic has high
activity against both of the
Staphylococcus aureus and Streptococcus
Pyogenes, while the Carbnicillin
antibiotic has high activity against of the
Bacillus spp, and when comparison the
activity of the all antibiotics types used
found of the Trimethoprim has high
activity against pathogenic gram positive
bacteria were used in this study.
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