Antimicrobial Sensitivity, Biochemical Characteristics and Biotyping of Staphylococcus saprophyticus: An Impact of Biofield Energy Treatment

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ISSN: 2167-0420
Women’s Health Care
Trivedi et al., J Women’s Health Care 2015, 4:6
http://dx.doi.org/10.4172/2167-0420.1000271
Volume 4 • Issue 6 • 1000271
J Women’s Health Care
ISSN: 2167-0420 JWHC, an open access journal
Open Access
Research Article
Keywords: Staphylococcus saprophyticus; Antimicrobial
susceptibility; Bioeld energy treatment; Biochemical reaction; Biotype;
Antibiogram; Gram-positive
Abbreviations: NIH/NCCAM: National Institute of Health/
National Center for Complementary and Alternative Medicine; ATCC:
American Type Culture Collection; PBPC 20: Positive Breakpoint
Combo 20; MIC: Minimum Inhibitory Concentration; CoNS:
Coagulase-negative staphylococci; UTIs: Urinary tract infections
Introduction
Staphylococcus saprophyticus (S. saprophyticus) is a Gram-positive,
coagulase-negative facultative bacterium belongs to Micrococcaceae
family. It is a unique uropathogen associated with uncomplicated
urinary tract infections (UTIs), especially cystitis in young women.
Young women are very susceptible to colonize this organism in
the urinary tracts and it is spread through sexual intercourse. S.
saprophyticus is the second most common pathogen aer Escherichia
coli causing 10-20% of all UTIs in sexually active young women [1-
3]. It contains the urease enzymes that hydrolyze the urea to produce
ammonia. e urease activity is the main factor for UTIs infection.
Apart from urease activity it has numerous transporter systems to
adjust against change in pH, osmolarity, and concentration of urea in
human urine [2]. Aer severe infections, it causes various complications
such as native valve endocarditis [4], pyelonephritis, septicemia, [5],
and nephrolithiasis [6]. About 150 million people are diagnosed with
UTIs each year worldwide [7]. Several virulence factors includes due
to the adherence to urothelial cells by release of lipoteichoic acid is a
surface-associated adhesion amphiphile [8], a hemagglutinin that
binds to bronectin and hemagglutinates sheep erythrocytes [9],
a hemolysin; and production of extracellular slime are responsible
for resistance properties of S. saprophyticus [1]. Based on literature,
S. saprophyticus strains are susceptible to vancomycin, rifampin,
gentamicin and amoxicillin-clavulanic, while resistance to other
antimicrobials such as erythromycin, clindamycin, uoroquinolones,
chloramphenicol, trimethoprim/sulfamethoxazole, oxacillin, and
Abstract
Staphylococcus saprophyticus (S. saprophyticus) is a frequent cause of urinary tract infection in the young women.
The current study was designed to analyze the effect of bioeld energy treatment on S. saprophyticus for evaluation
of its antibiogram prole, biochemical reactions pattern and biotyping characteristics. Two sets of ATCC samples
were taken in this experiment and denoted as A and B. Sample A was revived and divided into two parts Group (Gr.I)
(control) and Gr.II (revived); likewise, sample B was labeled as Gr.III (lyophilized). Gr. II and III were given with Mr.
Trivedi’s bioeld energy treatment. The control and treated groups of S. saprophyticus cells were tested with respect to
antimicrobial susceptibility, biochemical reactions pattern and biotype number using MicroScan Walk-Away® system.
The 50% out of twenty-eight tested antimicrobials showed signicant alteration in susceptibility and 36.67% out of thirty
antimicrobials showed an alteration in minimum inhibitory concentration (MIC) value of S. saprophyticus in revived
treated cells (Gr. II, day 10), while no alteration was found in lyophilized treated cells (Gr. III, day 10) as compared to
the control. It was also observed that overall 14.81%, out of twenty-seven biochemical reactions were altered in the
revived treated group with respect to the control. Moreover, biotype number was changed in Gr. II, on day 5 (246076)
and in Gr. III, on day 10 (242066), while organism along-with biotype number was also changed in Gr. II, on day 10
(342066, Staphylococcus hominis subsp. novobiosepticus) as compared to the control (242076, S. saprophyticus).
The result suggested that bioeld treatment has the signicant impact on S. saprophyticus in revived treated cells with
respect to the antimicrobial susceptibility, MIC, biochemical reactions pattern and biotype.
penicillin [10]. An alternative i.e., bioeld energy based healing therapy
is recently reported to alter the antimicrobial sensitivity pattern in
dierent microorganisms [11]. Bioeld (putative energy elds) or
electromagnetic based energy therapies, used to promote health and
healing had been exclusively reported by National Institute of Health/
National Center for Complementary and Alternative Medicine (NIH/
NCCAM) [12]. e human body naturally emits the waves in the form
of bio-photons, which surrounds the body and it is commonly known as
bioeld. In the recent year, Prakash et al. reported that various scientic
instruments such as Kirlian photography, polycontrast interference
photography and resonance eld imaging can be extensively used to
measure the bioeld of human body [13]. Although, a human has the
capability to harness the energy from the environment or universe and
can transmit it into any object(s) around the Globe. e objects always
receive the energy and responding in a useful way that is called bioeld
energy and the process is called as bioeld treatment. Mr. Trivedi’s
unique bioeld energy treatment (e Trivedi Eect®) has been known
to alter the characteristics features of pathogenic microbes [14,15], an
improved growth and productivity of plants [16,17] and also able to alter
the thermophysical properties of metal and ceramic in materials science
[18,19]. Due to the clinical importance of S. saprophyticus and literature
reports on bioeld, this work was undertaken to evaluate the impact of
*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: 917556660006; E-mail: [email protected]
Received September 21, 2015; Accepted September 23, 2015; Published
October 01, 2015
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al.
(2015) Antimicrobial Sensitivity, Biochemical Characteristics and Biotyping
of Staphylococcus saprophyticus: An Impact of Bioeld Energy Treatment. J
Women’s Health Care 4: 271. doi:10.4172/2167-0420.1000271
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.
Antimicrobial Sensitivity, Biochemical Characteristics and Biotyping of
Staphylococcus saprophyticus
: An Impact of Biofield Energy Treatment
Mahendra Kumar Trivedi1, Alice Branton1, Dahryn Trivedi1, Gopal Nayak1, 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
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al. (2015) Antimicrobial Sensitivity, Biochemical Characteristics and Biotyping of
Staphylococcus saprophyticus: An Impact of Bioeld Energy Treatment. J Women’s Health Care 4: 271. doi:10.4172/2167-0420.1000271
Page 2 of 5
Volume 4 • Issue 6 • 1000271
J Women’s Health Care
ISSN: 2167-0420 JWHC, an open access journal
bioeld treatment in relation to the antimicrobials susceptibility and
biotyping based on various biochemical characteristics.
Materials and Methods
S. saprophyticus, American Type Culture Collection (ATCC
15305) strains were procured from MicroBioLogics, Inc., USA, in
two sets A and B. e antimicrobials and biochemicals were used
in this experiment procured from Sigma-Aldrich, MA, USA. e
antimicrobial susceptibility, biochemical reaction pattern and biotype
number were estimated with the help of MicroScan Walk-Away® (Dade
Behring Inc., West Sacramento, CA, USA) using Positive Breakpoint
Combo 20 (PBPC 20) panel.
Experimental Design
Two ATCC 15305 samples A and B of S. saprophyticus were grouped
(Gr.). ATCC A sample was revived and divided into two parts named as
Gr.I (control) and Gr.II (revived, treated); likewise, ATCC B was labeled
as Gr.III (lyophilized, treated).
Bioeld Treatment Strategy
e control sample (Gr. 1) was remained as untreated. e treated
groups, Gr. II and III were handed over in sealed pack to Mr. Trivedi
for bioeld energy treatment under laboratory conditions. Mr. Trivedi
provided the treatment through his energy transmission process to
the treated groups (Gr. II and Gr. III) without touching the samples.
Aer treatment, all treated samples were stored for analysis in the
same condition. Gr.II was assessed at two time point i.e. on day 5
and 10 and Gr. III was assessed on day 10 aer the bioeld treatment,
for antimicrobial susceptibility, biochemical reactions pattern and
biotyping.
Antimicrobial Susceptibility Test
e antimicrobial susceptibility of S. saprophyticus was carried
out with the help of automated instrument, MicroScan Walk-Away®
using PBPC 20 panel. e panel was allowed to equilibrate to room
temperature before rehydration. All opened panels were used on the
same day. e tests carried out on MicroScan were miniaturized of the
broth dilution susceptibility test that has been dehydrated. Briey, 0.1
mL of the standardized suspension of S. saprophyticus cultured cells were
taken into 25 mL of inoculum water using pluronic and inverted 8 to 10
times and inoculated, rehydrated, and then subjected to incubation for
16 hours at 35°C. Aer that, rehydration and followed by inoculation
were performed using the RENOK® system with inoculators-D (B1013-
4). Approximately 25 mL of standardized inoculum suspension was
poured into the inoculum tray. e detailed experimental procedure
and conditions were maintained as per the manufacturer’s instructions.
e antibiogram prole like as susceptible, resistant, β-lactamase
positive (BLAC) and minimum inhibitory concentration (MIC) were
determined [20].
Biochemical Reaction Studies
e biochemical reactions of S. saprophyticus were determined
using MicroScan Walk-Away® system with PBPC 20 panel. Preparation
of PBPC 20 panel, inoculum and followed by dehydration and
rehydration were performed in a similar way as mentioned in the
antimicrobial susceptibility assay for the analysis of biochemical
reactions followed by biotype number. e MicroScan Walk-Away®
system contains photometric or uorogenic reader. On the basis
of nature of bacilli (i.e. Gram-positive), computerized reports were
generated using conventional panel, which utilizes the photometric
reader. Before commencing the experiment, the PBPC 20 panel was
rst incubated and read on the MicroScan Walkaway system. Aer
evaluating the experimental reading on the Walkaway system, the
PBPC 20 panel was removed from the system and recorded on the
Biomic system within 1 h. e instrument consists of a database
associated with collective information, which was required to identify
the microbes with respect to group, genera, or species of the family.
Detailed experimental procedure was followed as per manufacturer-
recommended instructions [20].
Identication of Organism By Biotype Number
e biotype number of S. saprophyticus was determined on
MicroScan Walk-Away® processed panel data report with the help
of biochemical reactions data. e similar experimental procedure
was followed for identication of biotype number as described in
biochemical reaction study, and as per manufacturer-recommended
instructions [20].
Results and Discussion
Antimicrobial susceptibility test
e data obtained in this experiment related to antimicrobials
sensitivity prole and MIC values are illustrated in Tables 1 and 2,
respectively. Aer bioeld energy treatment, the data were analyzed
and compared with respect to the control. e study was carried out
S. No Antimicrobial Gr. I Gr. II Gr. III
(Day 10)
Day 5 Day 10
1. Amoxicillin/k-clavulanate S S R S
2. Ampicillin/sulbactam S S R S
3. Ampicillin S S BLAC S
4. Azithromycin S S S S
5. Cefazolin S S R S
6. Cefepime S S R S
7. Cefotaxime S S R S
8. Ceftriaxone S S R S
9. Cephalothin S S R S
10. Chloramphenicol S S S S
11. Ciprooxacin S S S S
12. Clindamycin S S R S
13. Erythromycin S S S S
14. Gatioxacin S S S S
15. Gentamicin S S S S
16. Imipenem S S R S
17. Levooxacin S S S S
18. Linezolid S S S
19. Moxioxacin S S S S
20. Ooxacillin S S S S
21. Oxacillin S S R S
22. Penicillin S S BLAC S
23. Piperacillin/tazobactam S S S
24. Rifampin S S S S
25. Synercid S S R S
26. Tetracycline S S S S
27. Trimethoprim/sulfamethoxazole S S S S
28. Vancomycin S S R S
R: Resistant; S: Susceptible; Gr.: Group; −, Data not available; BLAC: β-lactamase
positive.
Table 1: Antibiogram of Staphylococcus saprophyticus: Effect of bioeld treatment
on antimicrobial susceptibility.
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al. (2015) Antimicrobial Sensitivity, Biochemical Characteristics and Biotyping of
Staphylococcus saprophyticus: An Impact of Bioeld Energy Treatment. J Women’s Health Care 4: 271. doi:10.4172/2167-0420.1000271
Page 3 of 5
Volume 4 • Issue 6 • 1000271
J Women’s Health Care
ISSN: 2167-0420 JWHC, an open access journal
with twenty-eight antimicrobials for assessment of susceptibility
assay and thirty antimicrobials for estimation of MIC. Several
antimicrobials viz. amoxicillin/k-clavulanate, ampicillin/sulbactam,
cefazolin, cefepime, cefotaxime, ceriaxone, cephalothin, clindamycin,
imipenem, oxacillin, synercid and vancomycin were converted
from susceptible (S) to resistance (R) in revived treated strain of S.
saprophyticus (Gr. II) on day 10, while S in Gr. II, on day 5 and in Gr. III
on day 10 as compared to the control (Gr. I). Moreover, ampicillin and
penicillin were changed the sensitivity pattern from S to β-lactamase
positive (BLAC) in Gr. II on day 10, while remained same i.e., S in Gr.
II, on day 5 and in lyophilized treated cells (Gr. III) on day 10 with
respect to the untreated group. Based on the literature, the species S.
saprophyticus has the ability to produce BLAC or penicillinase enzyme
that breakdowns the β-lactam ring present in penem heteronucleus
[21]. In this experiment, the enzyme production ability might be
enhanced aer bioeld energy treatment in revived treated cells. Hence,
these two penem containing antibiotics showed BLAC response in
Gr. II that was susceptible in the control sample. In this experiment,
authors have found that aer bioeld treatment the organism could
be able to produce β-lactamase enzyme that breaks the β-lactam ring
present in either penems or cephems nucleus. Overall, the treated cells
of S. saprophyticus showed a signicant (50%) alteration (fourteen
out of twenty-eight) in antimicrobial sensitivity pattern in the revived
treated Gr. II on day 10 as compared with the control. Fourteen, out of
twenty-eight tested antimicrobials did not show any alteration in the
sensitivity prole of the treated cells of S. saprophyticus. e MIC values
had revealed that various antibiotics such as ampicillin/sulbactam,
cefazolin, imipenem, linezolid, and synercid were altered by two-fold
in the revived treated group as compared to the control. Moreover, the
MIC values of vancomycin and oxacillin were changed aer bioeld
energy treatment by eight-fold in Gr. II on day 10 as compared to the
control. e alterations in MIC values were observed by several-fold
in ampicillin, clindamycin, oxacillin, and penicillin, in Gr. II on day
10 as compared to the control. Amoxicillin/k-clavulanate showed
slight alteration in MIC in Gr. II on day 10 while unaltered in rest of
treated groups with respect to the control. Overall, 36.67% (eleven out
of thirty) antimicrobials exhibited an alteration in MIC value in revived
treated cells (Gr. II, day 10) and rest of the antimicrobials did not report
any change in MIC values in all the treated groups as compared to the
control (Table 2). To achieve a strain-specic targeted drug therapy
for UTIs, in vitro patient-specic data are necessary because, the data
had provided wide geographical variability [22-24]. e production
of β-lactamase is common among staphylococci and is the prime
mechanism of penicillin resistance by these organisms. However, some
researchers unable to nd the enzyme that are responsible for resistance
among the strain of S. saprophyticus [25,26].
Biochemical reactions studies
e study of biochemical reactions are the test battery for
phenotypic identication of coagulase-negative staphylococci
(CoNS). e specic biochemical that showed some changes against
S. saprophyticus aer bioeld treatment as shown in Table 3. Based on
S. No. Antimicrobial Gr. I Gr. II Gr. III
(Day 10)
Day 5 Day 10
1. Amoxicillin/k-clavulanate ≤4/2 ≤4/2 >4/2 ≤4/2
2. Ampicillin/sulbactam ≤8/4 ≤8/4 >16/8 ≤8/4
3. Ampicillin ≤0.25 ≤0.25 4 ≤0.25
4. Azithromycin ≤2 ≤2 ≤2 ≤2
5. Cefazolin ≤8 ≤8 16 ≤8
6. Cefepime ≤8 ≤8 ≤8 ≤8
7. Cefotaxime ≤8 ≤8 ≤8 ≤8
8. Ceftriaxone ≤8 ≤8 ≤8 ≤8
9. Cephalothin ≤8 ≤8 ≤8 ≤8
10 Chloramphenicol ≤8 ≤8 ≤8 ≤8
11. Ciprooxacin ≤1 ≤1 ≤1 ≤1
12. Clindamycin ≤0.5 ≤0.5 >2 ≤0.5
13. Erythromycin ≤0.5 ≤0.5 ≤0.5 ≤0.5
14. Gatioxacin ≤2 ≤2 ≤2 ≤2
15. Gentamicin ≤4 ≤4 ≤4 ≤4
16. Imipenem ≤4 ≤4 >8 ≤4
17. Levooxacin ≤2 ≤2 ≤2 ≤2
18. Linezolid ≤2 ≤2 >4 ≤2
19. Moxioxacin ≤2 ≤2 ≤2 ≤2
20. Nitrofurantoin ≤32 ≤32 ≤32 ≤32
21. Noroxacin ≤4 ≤4 ≤4 ≤4
22. Ooxacin ≤2 ≤2 ≤2 ≤2
23. Oxacillin ≤0.25 ≤0.25 2 ≤0.25
24. Penicillin 0.12 0.12 >8 0.12
25. Piperacillin/tazobactam ≤4 ≤4 ≤4
26. Rifampin ≤1 ≤1 ≤1 ≤1
27. Synercid ≤1 ≤1 >2 ≤1
28. Tetracycline ≤4 ≤4 ≤4 ≤4
29. Trimethoprim/
sulfamethoxazole ≤2/38 ≤2/38 ≤2/38 ≤2/38
30. Vancomycin ≤2 ≤2 >16 ≤2
MIC data are presented in µg/mL; Gr: Group; −, Data not available
Table 2: Effect of bioeld treatment on Staphylococcus saprophyticus to minimum
inhibitory concentration (MIC) value of tested antimicrobials.
S. No. Code Biochemical Gr. I Gr. II Gr. III
(Day 10)
Day 5 Day 10
1. ARA Arabinose - - - -
2. ARG Arginine - - - -
3. BAC Bacillosamine + + + +
4. BE Bile esculin - - - -
5. CV Crystal violet - - - -
6. IDX Indoxyl phosphatase - - - -
7. INU Inulin - - - -
8. LAC Acidication Lactose + + + +
9. MAN Mannitol + + - -
10. MNS Mannose - - - -
11. MS Micrococcus screen + + + +
12. NACL Sodium chloride + + + +
13. NIT Nitrate - - +-
14. NOV Novobiocin + + + +
15. OPT Optochin + + + +
16. PGR Glycosidase*- - - -
17. PGT Glycosidase#+ + + +
18. PHO Phosphatase - - - -
19. PRV Pyruvate - - - -
20. PYR Pyrolidonyl arylamidase - - - -
21. RAF Rafnose - - - -
22. RBS Rambose - - - -
23. SOR Sorbitol - - - -
24. TFG Thymidine free growth + + + +
25. TRE Acidication trehalose + + -+
26. URE Urea + + + +
27. VP Voges-Proskauer - +- -
-: Negative; +: Positive; Gr: Group; *PGR: p-nitro phenyl β-D- glucuronide; #PGT:
p-nitro phenyl β-D-galactopyranoside
Table 3: Effect of bioeld treatment on Staphylococcus saprophyticus to the
biochemical reaction pattern.