Antimicrobial Susceptibility Pattern, Biochemical Characteristics and Biotyping of Salmonella paratyphi A: An Impact of Biofield Treatment

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Antimicrobial Susceptibility Pattern, Biochemical Characteristics and Biotyping
of Salmonella paratyphi A: An Impact of Biofield Treatment
Mahendra Kumar Trivedi1, Alice Branton1, Dahryn Trivedi1, Gopal Nayak1, Harish Shettigar1, Sambhu Charan Mondal2 and Snehasis Jana2*
1Trivedi Global Inc., 10624 S Eastern Avenue Suite A-969, Henderson, NV 89052, USA
2Trivedi Science Research Laboratory Pvt. Ltd., Hall-A, Chinar Mega Mall, Chinar Fortune City, Hoshangabad Rd., Bhopal-462026, Madhya Pradesh, India
*Corresponding author: Snehasis Jana, Trivedi Science Research Laboratory Pvt. Ltd., Hall-A, Chinar Mega Mall, Chinar Fortune City, Hoshangabad Rd.,
Bhopal-462026, Madhya Pradesh, India, Tel: +91-755-6660006; E-mail: [email protected]
Received date: July 20, 2015; Accepted date: August 13, 2015; Published date: August 20, 2015
Copyright: © 2015 Trivedi MK, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Enteric fever is a major global problem. Emergence of antimicrobial resistance threatens to render current
treatments ineffective. The current study was attempted to investigate the effect of biofield treatment on Salmonella
paratyphi A (S. paratyphi A) in terms of antimicrobial susceptibility assay, biochemical characteristics and biotyping.
S. paratyphi A strain were procured from MicroBioLogics in sealed packs bearing the American Type Culture
Collection (ATCC 9150). The study was conducted in revived and lyophilized state of S. paratyphi A. Both revived
(Group; Gr. II) and lyophilized (Gr. III) strain of S. paratyphi A were subjected to Mr. Trivedi’s biofield treatment.
Revived treated cells was assessed on day 5 and day 10, while lyophilized treated cells assessed on day 10 after
biofield treatment with respect to control (Gr. I). The antimicrobial susceptibility of S. paratyphi A showed significant
(60%) alteration in revived treated cells (Gr. II) on day 10 as compared to control. The MIC values of S. paratyphi A
also showed significant (53.12%) alteration in Gr. II and on day 10 while, no alteration was found in Gr. on day 5 as
compared to control. It was observed that overall 18.18% biochemical reactions were altered in the treated groups
with respect to control. Moreover, biotype numbers were substantially changed in Gr. II, on day 5 (53001040, S.
paratyphi A), on day 10 (57101050, Citrobacter freundii complex) as compared to control (53001000, S. paratyphi
A). Besides, biotype number was also changed in Gr. III (53001040, S. paratyphi A) as compared to control. The
overall result suggested that biofield treatment had significant impact on S. paratyphi A in Gr. II on day 10 with
respect to antimicrobial susceptibility, MIC values and biotype number.
Keywords:
S. paratyphi
A; Paratyphoid fever; Antimicrobial
susceptibility; Biofield treatment; Biochemical reaction; Biotype
Abbreviations
MIC: Minimum Inhibitory Concentration; ATCC: American Type
Culture Collection; NBPC 30: Negative Breakpoint Combo 30
Introduction
Enteric fever is a major public health problem in India.
Salmonella
enterica
(
S. enterica
) is motile, non-lactose fermenting, non-spore
forming, and Gram-negative rod shape bacterium. It can ferment
glucose with production of acid and gas. The subspecies of
enterica
are
three serotypes such as
paratyphi
A, B, and C that cause paratyphoid
fever [1]. The important virulence factor of
Salmonella paratyphi
A (
S.
paratyphi
A) is cytolysin A i.e. a cytotoxic protein which form pores in
the cell membrane [2]. Salmonellae mainly causes infection through
contaminated food or drink to the gut first through this protein then
enter into bloodstream. After that, it can infect to other parts of the
body. Manifestation of clinical symptoms such as diarrhoea and rashes
from paratyphoid fever usually occur between 6-30 days after being
infected [3].
S. enterica
serovar paratyphi A, B, or C are estimated to
cause 5.5 million cases of enteric fever each year [4]. In the United
States, paratyphoid fever is uncommon, while, an estimated 5.4 million
outbreaks occur in East Asia in 2000 [5].
Fluoroquinolone derivatives like ciprofloxacin, nalidixic acid etc.
are extensively used to combat paratyphoid fever in most parts of
world. In East Asian countries like India the prevalence of this disease
is too high. In India, newer cephalosporins derivatives, azithromycin,
chloramphenicol and ofloxacin are mostly prescribed against
paratyphoid fever, due to development of resistance towards
ciprofloxacin and nalidixic acid [6-8]. The patients with paratyphoid
fever especially in immunocompromised cases have a high rate of
relapse. Due to nature of this phenomena a short course treatment
with either fluoroquinolones or newer cephalosporins derivatives are
ineffective. There are not much data available in this area to provide
satisfactory treatment recommendations. Antibiotics are used to kill
the bacteria, but with increasing rates of antibiotic resistance, this
treatment is becoming less effective. Delay in receiving appropriate
antibiotic treatment may have serious consequences that lead to high
rates of mortality [9]. Therefore, some alternative strategies are needed
to treat against strains of
S. paratyphi
A.
Harold Saton Burr, had performed the detailed studies on the
correlation of electric current with physiological process and
concluded that every single process in the human body had an
electrical significance [10]. Recently, it was discovered that all the
electrical process happening in body have strong relationship with
magnetic field as described by Ampere’s law, which states that the
moving charge produces magnetic fields in surrounding space [11,12].
According to Rivera-Ruiz et al. reported that electrocardiography has
been extensively used to measure the biofield of human body [13].
Thus, human has the ability to harness the energy from environment
or universe and can transmit into any living or nonliving object(s)
around the Globe. The objects always receive the energy and
Clinical Microbiology: Open Access Trivedi et al., Clin Microbiol 2015, 4:4
http://dx.doi.org/10.4172/2327-5073.1000215
Research Article Open Access
Clin Microbiol
ISSN:2327-5073 CMO, an open access journal Volume 4 • Issue 4 • 1000215
responding into useful way that is called biofield energy and the
process is known as biofield treatment. Mr. Trivedi’s unique biofield
treatment is also known as The Trivedi Effect®. Mr. Mahendra
Trivedi’s biofield treatment has been known to transform the
structural, physical and thermal properties of several metals in
material science [14-16], improved the overall productivity of crops
[17,18], altered characteristics features of microbes [19-21] and
improved growth and anatomical characteristics of various medicinal
plants [22,23].
Due to the clinical significance of this organism and literature
reports on biofield treatment as an alternative approach, the present
work was undertaken to evaluate the impact of biofield treatment on
S.
paratyphi
A in relation to antimicrobials susceptibility, minimum
inhibitory concentration (MIC) and biotyping based on various
biochemical characters.
Materials and Methods
S. paratyphi
A, American Type Culture Collection (ATCC 9150)
strains were procured from MicroBioLogics, Inc., USA, in two sets A
and B. Two different sealed packs were stored with proper storage
conditions until further use. The antimicrobial susceptibility, MIC
values, biochemical reactions and biotype number were estimated with
the help of MicroScan Walk-Away® (Dade Behring Inc., West
Sacramento, CA, USA) using negative breakpoint combo 30 (NBPC
30) panel with respect to control sample. All the tested antimicrobials
and biochemicals were procured from Sigma-Aldrich (MA, USA).
Experimental design
Two ATCC samples A (revived) and B (lyophilized) of
S. paratyphi
A were grouped (Gr.). The revived sample A was divided into two
parts Gr.I (control) and Gr.II (revived; treatment); likewise, ATCC B
was labeled as Gr.III (lyophilized; treatment).
Biofield treatment strategy
Gr. I remained as untreated. The treatment Gr. II and III in sealed
pack were handed over to Mr. Trivedi for biofield treatment under
laboratory condition. Mr. Trivedi provided the treatment through his
energy transmission process to the treatment groups without touching
the samples. After treatment, all samples were handed over in the same
condition and stored for analysis. Gr.II was assessed at two time point
i.e. on day 5 and 10 and Gr. III was assessed on day 10. After biofield
treatment, all the groups (control and treated) were investigated for
antimicrobial susceptibility, MIC, biochemical reactions pattern and
biotyping.
Antimicrobial susceptibility test
Investigation of antimicrobial susceptibility of
S. paratyphi
A was
carried out with the help of automated instrument, MicroScan Walk-
Away® using NBPC 30 panel. The panel can be stored at 2°C-25°C for
analysis. The panel was allowed to equilibrate to room temperature
prior to rehydration. All opened panel was used within the same day.
The tests carried out on MicroScan were miniaturized of the broth
dilution susceptibility test that has been dehydrated. Briefly, the 0.1 ml
(100 μl) of the standardized suspension of
S. paratyphi
A was pipetted
into 25 ml of inoculum water using pluronic and inverted 8-10 times
and inoculated, rehydrated, and then subjected to incubation for 16
hours at 35°C. Rehydration and inoculation was performed using the
RENOK® system with inoculators-D (B1013-4). 25 ml of standardized
inoculum suspension was poured in to inoculum tray. The detailed
experimental procedures and conditions were followed as per the
manufacturer's instructions. The antimicrobial susceptibility pattern
(S: Susceptible, R: Resistant; and I: Intermediate) and MIC values were
determined by observing the lowest antimicrobial concentration
showing inhibition of growth [24].
Biochemical reaction studies
Biochemical activities of
S. paratyphi
A was determined using
MicroScan Walk-Away®, system with NBPC 30 panel. Preparation of
NBPC 30 panel, inoculum followed by dehydration and rehydration
were performed similar way as mentioned in antimicrobial
susceptibility assay for analysis of biochemical reaction followed by
biotype number. The detailed experimental procedures and conditions
were followed as per the manufacturer's instructions [24,25].
Identification of organism by biotype number
The biotype number of
S. paratyphi
A was determined on
MicroScan Walk-Away® processed panel data report with the help of
biochemical reactions data. [24,26].
Results and Discussion
Antimicrobial susceptibility test
The outcome of
S. paratyphi
A susceptibility pattern and MIC
values of tested antimicrobials after biofield treatment are summarized
in Tables 1 and 2 respectively. The data were analyzed and compared
with respect to control. Antimicrobial sensitivity was carried out in
twenty antimicrobials. The revived treated cells (Gr. II) of
S. paratyphi
A showed a significant (60%) alteration (twelve out of twenty) in
antimicrobial sensitivity pattern on day 10 while did not show any
alteration on day 5 as well as in Gr. III as compared to control.
Antimicrobials such as amoxicillin/k-clavulanate, ampicillin/
sulbactam, aztreonam, chloramphenicol, piperacillin, tetracycline and
ticarcillin/k-clavulanate showed alteration of susceptibility pattern
from S to I in Gr. II on day 10 as compared to control. Moreover,
antimicrobial agents such as ampicillin, ceftazidime, ceftriaxone,
ciprofloxacin and trimethoprim/sulfamethoxazole showed alteration
of sensitivity pattern from S to R in Gr. II on day 10 as compared to
control after biofield treatment. Eight out of twenty (40%)
antimicrobials did not show any change of antimicrobial sensitivity
after biofield treatment with respect to control sample (Table 1). The
MIC values of amoxicillin/k-clavulanate and ampicillin/sulbactam
were changed from ≤ 8/4 µg/ml to 16/8 µg/ml in Gr. II on day 10.
Moreover, MIC values of ampicillin and ceftazidime were also
changed from ≤ 8 µg/ml to >16 µg/ml) in Gr. II on day 10 as compared
to control. Alteration of MIC values (≤ 8 µg/ml to 16 µg/ml) were
noticed in revived treated samples of certain antimicrobials like
aztreonam, cefoxitin, and chloramphenicol except cefuroxime were
changed from ≤ 4 µg/ml to 16 µg/ml on day 10. Moreover, change in
MIC values (≤ 4 µg/ml to 8 µg/ml) were observed in Gr. II in case of
norfloxacin, tetracycline and tobramycin on day 10 after biofield
treatment. Antimicrobials showed an alteration of MIC value (≤ 8
µg/ml to >32 µg/ml) in case of ceftriaxone, (≤ 1 µg/ml to >2 µg/ml) in
case of ciprofloxacin, (≤ 16 µg/ml to 64 µg/ml) in case of piperacillin
and ticarcillin/k-clavulanate, (≤ 2/38 µg/ml to >2/38 µg/ml) in case of
trimethoprim/sulfamethoxazole on day 10 in Gr. II. Besides this,
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Shettigar H, et al. (2015) Antimicrobial Susceptibility Pattern, Biochemical Characteristics
and Biotyping of Salmonella paratyphi A: An Impact of Biofield Treatment. Clin Microbiol 4: 215. doi:10.4172/2327-5073.1000215
Page 2 of 7
Clin Microbiol
ISSN:2327-5073 CMO, an open access journal Volume 4 • Issue 4 • 1000215
ESBL-b Scrn showed an alteration of MIC value (≤ 1 to >1 µg/ml) in
Gr. II on day 10. Overall, 53.12% (seventeen out of thirty two) MIC
values were altered after biofield treatment as compared to control.
Fifteen, out of thirty two tested antimicrobials (46.88%) did not show
any alteration of MIC values in treated cells of
S. paratyphi
A as
compared to control (Table 2). Overall, the antimicrobial resistance
pattern (S to I/R) and corresponding MIC values were significantly
altered in revived strain
S. paratyphi
A after biofield treatment as
compared to control. In this experiment, ciprofloxacin and cefotaxime
were sensitive to control
S. paratyphi
A (Gr. 1) and the MIC value was
slight increase in ciprofloxacin (≤ 1 µg/ml to >2 µg/ml) while, it did
not alter in case of cefotaxime in Gr.II on day 10. This finding is
supported by literature data [27].
S. No. Antimicrobial
Type of Response
Gr. I Gr. II Gr. III
Day 5 Day 10
1 Amoxicillin/k-clavulanate S S I S
2 Ampicillin/sulbactam S S I S
3 Ampicillin S S R S
4 Aztreonam S S I S
5 Cefepime S S S S
6 Cefotaxime S S S S
7 Ceftazidime S S R S
8 Ceftriaxone S S R S
9 Chloramphenicol S S I S
10 Ciprofloxacin S S R S
11 Gatifloxacin S S S S
12 Imipenem S S S S
13 Levofloxacin S S S S
14 Meropenem S S S S
15 Moxifloxacin S S S S
16 Piperacillin/tazobactam S S S S
17 Piperacillin S S I S
18 Tetracycline S S I S
19 Ticarcillin/k-clavulanate S S I S
20 Trimethoprim/sulfamethoxazole S S R S
R: Resistant; I: Intermediate; S: Susceptible; Gr.: Group
Table 1: Antibiogram of
Salmonella paratyphi A
: effect of biofield treatment on antimicrobial susceptibility.
S. No. Antimicrobial
Type of Response
Gr. III
Gr. I Gr. II
Day 5 Day 10
1 Amikacin ≤ 16 ≤ 16 ≤ 16 ≤ 16
2 Amoxicillin/k-clavulanate ≤ 8/4 ≤ 8/4 16/8 ≤ 8/4
3 Ampicillin/sulbactam ≤ 8/4 ≤ 8/4 16/8 ≤ 8/4
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Shettigar H, et al. (2015) Antimicrobial Susceptibility Pattern, Biochemical Characteristics
and Biotyping of Salmonella paratyphi A: An Impact of Biofield Treatment. Clin Microbiol 4: 215. doi:10.4172/2327-5073.1000215
Page 3 of 7
Clin Microbiol
ISSN:2327-5073 CMO, an open access journal Volume 4 • Issue 4 • 1000215