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Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of Biofield Treated Nocardia otitidis

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  • Trivedi Global, Inc

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Nocardiosis is a soil-borne aerobic infection caused by Nocardia species commonly affects the respiratory tract. Nocardia otitidis (N. otitidis) is the key organism for non-mycobacterial tuberculosis. The current study was attempted to investigate the effect of Mr. Trivedi’s biofield energy treatment on N. otitidis and analyzed for antimicrobial susceptibility pattern, minimum inhibitory concentration (MIC), DNA polymorphism by Random Amplified Polymorphic DNA (RAPD) and 16S rDNA sequencing. The strain of N. otitidis (ATCC 14630) was divided into two parts, control and treated. Antimicrobial susceptibility was studied using the broth microdilution technique. Overall, the MIC values of 16.67% antimicrobials were changed in the treated group of N. otitidis as compared to the control. Moreover, MIC value of trimethoprim/sulfamethoxazole was reduced by two-fold (0.5/9.5 to 0.25/4.75 µg/mL) in the biofield energy treated sample as compared to the control without alteration in the sensitivity spectrum. The 16S rDNA analysis showed that the treated sample was detected as Enterobacter aerogenes strain NCTC10006T (GenBank Accession No: AJ251468) with 98% identity of gene sequencing data. However, the nearest homolog genus-species was found as Kluyvera cryocrescens (GenBank Accession No: AM184245). Using RAPD biomarkers, the sample showed an average range of 34 to 53% of polymorphism among treated samples as compared to the control. The 16S rDNA sequencing of treated sample was carried out to correlate the phylogenetic relationship of N. otitidis with other bacterial species. These results suggested that Mr. Trivedi’s biofield energy treatment has a significant impact on N. Otitidis.
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ISSN: 2329-6577
Biological Systems: Open Access
Trivedi et al,, Biol Syst Open Access 2015, 4:2
http://dx.doi.org/10.4172/2329-6577.1000143
Open Access
Research Article
Volume 4 • Issue 2 • 1000143
Biol Syst Open Access
ISSN: 2329-6577 BSO, an open access journal
Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of
Biofield Treated
Nocardia otitidis
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
Abstract
Nocardiosis is a soil-borne aerobic infection caused by Nocardia species commonly affects the respiratory tract.
Nocardia otitidis (N. otitidis) is the key organism for non-mycobacterial tuberculosis. The current study was attempted to
investigate the effect of Mr. Trivedi’s bioeld energy treatment on N. otitidis and analyzed for antimicrobial susceptibility
pattern, minimum inhibitory concentration (MIC), DNA polymorphism by Random Amplied Polymorphic DNA (RAPD)
and 16S rDNA sequencing. The strain of N. otitidis (ATCC 14630) was divided into two parts, control and treated.
Antimicrobial susceptibility was studied using the broth microdilution technique. Overall, the MIC values of 16.67%
antimicrobials were changed in the treated group of N. otitidis as compared to the control. Moreover, MIC value of
trimethoprim/sulfamethoxazole was reduced by two-fold (0.5/9.5 to 0.25/4.75 µg/mL) in the bioeld energy treated
sample as compared to the control without alteration in the sensitivity spectrum. The 16S rDNA analysis showed that
the treated sample was detected as Enterobacter aerogenes strain NCTC10006T (GenBank Accession No: AJ251468)
with 98% identity of gene sequencing data. However, the nearest homolog genus-species was found as Kluyvera
cryocrescens (GenBank Accession No: AM184245). Using RAPD biomarkers, the sample showed an average range of
34 to 53% of polymorphism among treated samples as compared to the control. The 16S rDNA sequencing of treated
sample was carried out to correlate the phylogenetic relationship of N. otitidis with other bacterial species. These
results suggested that Mr. Trivedi’s bioeld energy treatment has a signicant impact on N. otitidis.
*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:
publication@trivedisrl.com
Received August 31, 2015; Accepted September 21, 2015; Published September
29, 2014
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al. (2015)
Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of Bioeld Treated
Nocardia otitidis. Biol Syst Open Access 4: 143. doi:10.4172/2329-6577.1000143
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.
nocardiosis [9] in immunocompromised patients. Infectious Diseases
Society of America (IDSA) reported that between 500 and 1,000 cases
of Nocardial infections are recognized in the United States each year, of
which 85% are serious pulmonary or systemic infections [10].
Since 1940s, the sulfonamides have been the drugs of choice for
the treatment of nocardiosis [11]. Due to high mortality rate (50%)
in patients with central nervous system (CNS) Nocardia infections
(Nocardia brain abscess) and patients with non-CNS overwhelming or
disseminated disease, the treatment strategy is inadequate. However, the
combination of sulfamethoxazole with trimethoprim is oen used as the
drug of choice for the treatment of nocardiosis [12]. erefore, some
alternative strategies are needed to treat against nocardiosis. Bioeld
energy has been known as an alternative approach which may be useful
as an alternative treatment to Nocardia infected patients. National
Institute of Health/National Center for Complementary and Alternative
Medicine (NIH/NCCAM) have reported that bioeld (putative energy
elds) or electromagnetic based energy therapies were commonly
used to promote the health and healing [13]. Harold Saxton Burr had
performed the detailed studies on the correlation of electric current
with the physiological process and concluded that every single process
in the human body had an electrical signicance [14]. Recently, it was
discovered that all the electrical processes happening in the human
body have strong relationship with the magnetic eld as required by
Keywords: Nocardia otitidis; Nocardiosis; Antimicrobial
susceptibility; Bioeld energy treatment; 16S rDNA sequencing;
Random amplied polymorphic DNA
Abbreviations: NIH/NCCAM: National Institute of Health/
National Center for Complementary and Alternative Medicine;
ATCC: American Type Culture Collection; MIC: Minimum Inhibitory
Concentration; OTUs: Operational Taxonomic Units; NCBI: National
Center for Biotechnology Information; MEGA: Molecular Evolutionary
Genetics Analysis; PCR: Polymerase Chain Reaction; RDP: Ribosomal
Database Project; RAPD: Random Amplied Polymorphic DNA; CNS:
Central Nervous System
Introduction
e genusNocardiais associated with the group of microorganisms
known as the aerobic actinomycetes and belongs to the family of
Mycobacteriaceae. Nocardia contains tuberculostearic acids but that
dier from the mycobacteria by the possession of short-chain (40 to
60 carbons) mycolic acids [1]. Nocardia otitidis (N. otitidis) is a weak
lamentous Gram-positive, catalase-positive, branching rods shaped
bacterium that appears similar to Actinomyces species. However, it
can usually be dierentiated from Actinomyces by acid-fast staining
[2]. e taxonomic history of the genus Nocardia is controversial [3].
Nocardia typically exhibits varying degrees of acid fastness due to the
presence of cell wall mycolic acid. e genus is typically similar to the
genus of Mycobacterium. Mordarska et al. had studied the short-chain
fatty acids content in the cell wall of Nocardia and Mycobacterium
genera based on gas-liquid chromatography analysis did not nd
any dierence between two genera [4]. Based on immunoblot and
enzyme-linked immunosorbent assay (ELISA) techniques that detect
specic antibodies that appear as common in various Nocardia and
Actinomadura species. ese antigens do not react with the antibodies
produced in response to Mycobacterium tuberculosis infections [5].
Most of Nocardial infections occur in the United States due to inhalation
of airborne spores or mycelial fragments from the environmental
sources [6]. e most common manifestation of Nocardial disease is
pulmonary nocardiosis [7], extrapulmonary disease [8], and ocular
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al. (2015) Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of
Bioeld Treated Nocardia otitidis. Biol Syst Open Access 4: 143. doi:10.4172/2329-6577.1000143
Page 2 of 6
Volume 4 • Issue 2 • 1000143
Biol Syst Open Access
ISSN: 2329-6577 BSO, an open access journal
Ampere’s law, which states that the moving charge produces magnetic
elds in the surrounding space [15,16]. us, a human body emits the
electromagnetic waves in the form of bio-photons that is also known
as ultra-weak photon emissions (UPE). It surrounds the body and it
is commonly known as bioeld. erefore, the bioeld consists of an
electromagnetic eld, being generated by moving electrically charged
particles (ions, cell, molecule, etc.) inside the human body [17]. e
transfer of information from cell to cell or DNA or storage by biophotons
has been demonstrated in plants, bacteria, animal neutriophil
granulocytes and kidney cells [18]. Prakash et al. in 2015 reported that the
various scientic instruments such as Kirlian photography, polycontrast
interference photography (PIP) and resonance eld imaging (RFI) can
be extensively used to measure the bioeld of human body [19]. us,
human has the ability to harness the energy from the environment or
universe and can transmit into any living or nonliving object(s) around
the Globe. e objects always receive the energy and responding into
the useful way that is called bioeld energy and the process is known as
bioeld energy treatment. Mr. Mahendra Trivedi’s unique bioeld energy
treatment (e Trivedi Eect®) has been known to improve the overall
productivity of crops [20,21], altered characteristics features of microbes
[22-24], alter the structural, physical and thermal properties of several
metals [25,26], and improved growth and anatomical characteristics of
various medicinal plants [27,28].
Based on clinical signicance of N. otitidis and signicant impact
of Mr. Trivedi’s bioeld energy modality on microbes, the present work
was undertaken to evaluate the impact of Mr. Trivedi’s bioeld energy
on N. otitidis in relation to antimicrobials susceptibility, minimum
inhibitory concentration (MIC), random amplied polymorphic DNA
analysis (RAPD) and 16S rDNA sequencing.
Materials and Methods
N. otitidis, American Type Culture Collection (ATCC 14630) strain
was procured from Bangalore Genei, Bangalore-India in two vials A
and B. Two dierent sealed packs were stored with proper storage
conditions until further use. All the tested antimicrobials were procured
from Sigma-Aldrich, India. e antimicrobial susceptibility and
MIC were estimated with the help of broth micro dilution technique
as per the Clinical Laboratory Standards Institute (CLSI) guidelines
document number M24-A [29]. e 16S rDNA sequencing and DNA
ngerprinting (RAPD) studies were carried out using Ultrapure
Genomic DNA Prep Kit; Cat KT 83 (Bangalore Genei, India).
Experimental design and bioeld treatment strategy
N. otitidis strain was divided into two groups, i.e., control and
treated. e treated group was in sealed pack and handed over to Mr.
Trivedi for bioeld energy treatment under laboratory conditions. Mr.
Trivedi provided the treatment through his energy transmission process
to the treated group without touching the sample. Aer treatment,
control and treated groups were assessed on day 10 for antimicrobial
susceptibility, and MIC. For RAPD analysis three inoculums (one for
control and the other two for treated named as treated A and B) were
prepared from N. otitidis samples. ese two bioeld treated samples
A and B were sub-cultured by taking 1% inoculum and inoculated to
fresh 5 ml medium and labeled as treatment A-1 and treatment B-1,
respectively. e result of treated sample was compared with respect to
the control. e 16S rDNA analysis was performed on bioeld treated
samples A and its subcultured sample A1.
Antimicrobial susceptibility test
e antimicrobial susceptibility testing is important for clinically
signicant species. e investigation of antimicrobial susceptibility of N.
otitidis was carried out with the help of broth micro dilution technique,
as per CLSI guidelines. Broth micro dilution was recommended
for isolates of rapidly growing mycobacteria (RGM) based on CLSI
published guidelines and recommendations for testing of non-
tuberculous mycobacteria (CLSI, M24-A, 2003) [29]. e detailed
experimental procedure and conditions were followed as per the
manufacturer’s instructions. e antimicrobial susceptibility pattern
(S: Susceptible, R: Resistant) and minimum inhibitory concentration
(MIC) were determined by observing the lowest antimicrobial
concentration showing inhibition of growth.
Random amplied polymorphic DNA (RAPD) analysis
For DNA ngerprinting (RAPD) analysis all the treated samples
(A, A1, B, and B1) were incubated at 37°C with 160 rpm for 18 h.
Subsequently, the cultures were spun down, pelleted at 5000 rpm at
40°C for 10 minutes and the genomic DNA was isolated for control
and treated samples using Genomic DNA Prep Kit (Bangalore Genei,
India). RAPD was performed with all samples of N. otitidis using ve
RAPD primers, which were labelled as RBA8A, RBA13A, RBA20A,
RBA10A and RBA15A were adopted from earlier studies. e PCR
mixture contained 2.5 μL each of buer, 4.0 mM each of dNTP, 2.5 μM
each of primer, 5.0 μL (approximately 20 ng) of each genomic DNA,
2U each of Taq polymerase, 1.5 μL of MgCl2 and 9.5 μL of water in
a total of 25 μL with the following PCR amplication protocol; initial
denaturation at 94°C for 7 min, followed by 8 cycles of denaturation at
94°C for 1 min, annealing at 35°C for 1 min, and extension at 72°C for 2
min; and 35 cycle of denaturation at 94°C for 1 min, annealing at 38°C
for 1 min, and extension at 72°C for 1.5 min; and the nal extension
at 72°C for 7 min. Amplied PCR products from all samples (control
and treated) were separated on 1.5% agarose gels at 75 volts, stained
with ethidium bromide and visualized under UV illumination [30]. e
percentage of polymorphism was calculated using following equation:
Percent polymorphism=A/B × 100
Where, A=Number of polymorphic bands in treated sample; and
B=Number of polymorphic bands in control.
Amplication and gene sequencing of 16S rDNA
Genomic DNA was isolated from N. otitidis cells (set A, sample coded
as 9A) using genomic purication kit, according to the manufacturer
instructions. 16S rDNA gene (~ 1.5 kb) fragment was amplied with the
help of high-delity polymerase chain reaction (PCR) using universal
primers; forward primer (5-AGAGTTTGATCCTGGCTCAG-3) and
reverse primer (3-ACGGTCATACCTTGTTACGACTT-5). Amplied
products were subjected to gel electrophoresis in 1.0% agarose gel,
stained with ethidium bromide and visualized under UV light in a gel
documentation unit (BioRad Laboratories, USA). e PCR amplied
fragment was puried from the agarose gel using a DNA gel extraction
kit. Sequencing of amplied product was done on a commercial basis
from Bangalore Genei, India. e 16S rDNA sequences obtained were
aligned and compared with the sequences stored in GenBank database
available from National Center for Biotechnology Information (NCBI)
using the algorithm BLASTn program. Multiple sequence alignment/
phylogenetic tree were established using MEGA3.1 molecular soware
[31].
Results and Discussion
Antimicrobial susceptibility test
Antimicrobial susceptibility pattern of Nocardia can vary from
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al. (2015) Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of
Bioeld Treated Nocardia otitidis. Biol Syst Open Access 4: 143. doi:10.4172/2329-6577.1000143
Page 3 of 6
Volume 4 • Issue 2 • 1000143
Biol Syst Open Access
ISSN: 2329-6577 BSO, an open access journal
species to species. e therapeutic eectiveness depends on the proper
identication of species in infected patients and on in vitro sensitivity
studies [32]. e antimicrobial sensitivity assay against Nocardia
species had been especially considered in refractory cases. A standard
test for antimicrobials sensitivity assessed by broth micro dilution and
with cation-supplemented Mueller-Hinton broth has been approved by
the National Committee for Clinical Laboratory Standards (NCCLS)
[29]. e outcome of N. otitidis susceptibility pattern and MIC values of
tested antimicrobials aer bioeld energy treatment are summarized in
Table 1 and 2 respectively. e data were analyzed and compared with
respect to the control. Study was carried out in twelve antimicrobials
for assessment of antimicrobial susceptibility and MIC value. e
treated cells of N. otitidis did not show any alteration with respect to
antimicrobial susceptibility pattern as compared to the control (Table
1). e susceptible nature of N. otitidis to amikacin, sulfamethoxazole,
and ciprooxacin and resistance pattern to ceriaxone, amoxicillin-
clavulanic acid, and imipenem were well corroborated with the
literature data [3]. Beside this, the MIC value of trimethoprim/
sulfamethoxazole was reduced by two-fold (0.5/9.5 to 0.25/4.75 µg/mL)
in bioeld energy treated sample as compared to the control sample.
Moreover, the MIC value was slightly reduced (>32/16 to 32/16 µg/mL)
in amoxicillin/k-clavulanate aer the bioeld therapy. Overall, 16.67%
out of twelve antimicrobials showed an alteration of MIC values. Rest
of the antimicrobials did not show any alteration of MIC values with
respect to the control sample. Based on the literature, a combination
of sulfa and antimalarial drugs, i.e., trimethoprim/sulfamethoxazole
are the drug of choice against Nocardia infections as compared to the
single sulfa drug [11,12]. In this experiment, the susceptibility nature
of trimethoprim/sulfamethoxazole was constant in both controls as
well as in treated sample, while the MIC value was reduced by two-
fold in treated sample aer Mr. Trivedi’s bioeld energy treatment.
Hence, authors assumed that this improvement of MIC value without
alteration of sensitivity pattern may be due to the eect of putative
energy transmit through bioeld healing (Table 2).
Random amplied polymorphic DNA (RAPD) analysis
e treated and control samples were identied on the basis of
their dierent and discriminative RAPD patterns. RAPD is a preferred
tool that is being used now days to correlate the genetic similarity
or mutations between species. e simplicity and wide applicability
of RAPD analysis mainly depend upon the use of short nucleotide
primers, which were unrelated to known DNA sequences of the target
organism [33]. DNA polymorphisms can be eciently detected using
the PCR primers and identify inter-strain variations among species in
treated samples [34]. e degree of relatedness and genetic mapping
can be correlated between similar or dierent treated sample species
[35].
e DNA ngerprinting by RAPD analysis using ve primers was
carried out on the control and treated samples. DNA ngerprinting by
RAPD analysis of the control and treated samples are shown in Figure 1,
and the polymorphic bands are marked by arrows. e RAPD patterns
of treated samples showed some unique and dissimilar patterns. DNA
polymorphism analyzed by the RAPD analysis was presented in Table 3.
e level of polymorphism between control and treated samples (A, A1,
B, and B1) are summarized in Table 4. e level of polymorphism was
found in an average range of 34 to 53% in treated samples as compared
to control in N. otitidis aer the bioeld treatment. e highest change
in DNA sequence was observed in treated groups with RBA 13A primer
as compared to the control; a negligible change was found in treated
group with RBA 8A primer as compared to the control (Table 3 and 4).
16S rDNA genotyping
e bacteria that are poorly dierentiated by conventional methods
needs molecular analysis method like 16S rDNA sequence [36]. is
molecular-based technique is a suitable tool for identication of most
1000
900
800
700
600
500
400
300
200
100
bp
Figure 1: Random amplied polymorphic-DNA fragment patterns of Nocardia
otitidis generated using ve RAPD primers, RBA 8A, RBA 13A, RBA 20A, RBA
10A and RBA 15A. 1: Control; 2: Treated A; 3: Treated A-1; 4: Treated B; 5:
Treated B-1; M: 100 bp DNA Ladder.
S. No. Antimicrobial Control Treatment
1. Linezolid S S
2. Clarithromycin R R
3. Amikacin S S
4. Cefoxitin R R
5. Ceftriaxone R R
6. Imipenem R R
7. Minocycline S S
8. Tobramycin S S
9. Ciprooxacin S S
10. Gatioxacin S S
11. Amoxicillin/k-clavulanate R R
12. Trimethoprim/sulfamethoxazole S S
R: Resistant; S: Susceptible; Control: ATCC strain of N. otitidis (without bioeld
energy treatment); Treatment: ATCC strain of N. otitidis (with Mr. Trivedi’s bioeld
energy treatment)
Table 1: Antimicrobial susceptibility pattern of antimicrobials against ATCC strain of
Nocardia otitidis after bioeld treatment on day 10 as per CLSI guidelines.
S. No. Antimicrobial Control Treatment
1. Linezolid 2.0 2.0
2. Clarithromycin >32.0 >32.0
3. Amikacin <1.0 <1.0
4. Cefoxitin 256.0 256.0
5. Ceftriaxone 64.0 64.0
6. Imipenem 64.0 64.0
7. Minocycline 1.0 1.0
8. Tobramycin 8.0 8.0
9. Ciprooxacin 4.0 4.0
10. Gatioxacin 0.12 0.12
11. Amoxicillin/k-clavulanate >32/16 32/16
12. Trimethoprim/sulfamethoxazole 0.5/9.5 0.25/4.75
CLSI: Clinical and Laboratory Standards Institute; MIC values are presented
as µg/mL; Control: ATCC strain of N. otitidis (without bioeld energy treatment);
Treatment: ATCC strain of N. otitidis (with Mr. Trivedi’s bioeld energy treatment)
Table 2: Minimum inhibitory concentration (MIC) of antimicrobials against ATCC
strain of Nocardia otitidis after bioeld energy treatment on day 10 as per CLSI
guidelines.
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al. (2015) Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of
Bioeld Treated Nocardia otitidis. Biol Syst Open Access 4: 143. doi:10.4172/2329-6577.1000143
Page 4 of 6
Volume 4 • Issue 2 • 1000143
Biol Syst Open Access
ISSN: 2329-6577 BSO, an open access journal
of the bacteria on their genus and/or species level by comparison
with databases in the public domain. Because, most of the bacteria
have possess small ribosomal subunit with species-specic variability
[37]. e 16S rDNA sequence was determined in treated samples of
N. otitidis and coded as 9A and 9A1 (sub cultured sample), which are
shown in Table 5. e alignment and comparison of the consensus
gene sequences were performed with the sequences stored in GenBank
database available from NCBI using the algorithm BLASTn program.
Based on nucleotide homology and phylogenetic analysis of
the microbe, the samples 9A and 9A1 (N. otitidis) were detected as
Enterobacter aerogenes (GenBank Accession Number: AJ251468) with
98% identity of gene sequencing data. e nearest homolog genus-
species of N. otitidis (9A and 9A1) was found as Kluyvera cryocrescens
(Accession No. AM184245). Some other close homologs of N. otitidis
were found from the alignment results as shown in Table 5. e distance
matrix based on nucleotide sequence homology data are presented in
Table 6. e phylogenetic tree was established using BLAST-Webpage
(NCBI). According to Table 6, ten dierent related bacterial species
of N. otitidis were selected as Operational Taxonomic Units (OTUs)
in order to investigate the phylogenetic relationship of N. otitidis.
ere were 1462 base-pair nucleotides of 16S rDNA gene sequences,
which were analyzed and multiple alignments were constructed using
ClustalW in MEGA3.1 [31]. e numbers of base substitutions per site
from pairwise distance analysis between sequences are shown in Table
6. All the results were based on the pairwise analysis of 11 sequences.
According to the data presented in Table 6, the lowest value of the
genetic distance from N. otitidis was 0.018 base substitutions per site.
is value is due to the distance between Enterobacter aerogenes and
Kluyvera cryocrescens. All pairwise distance analysis was carried out
using the p-distance method in MEGA3.1. e proportion of remarked
distance, sometimes also called p-distance and showed as the number
of nucleotide distances site. Values in Table 6 are programmed into
Figure 2 with optimal bootstrap consensus tree. In the phylogram, there
were eleven OTUs. e results suggested that aer bioeld treatment
S. No. Primer Nucleotide Sequence
(5′-3′) Band Scored Common Bands in
Control and Treated
Unique Band
Control TSA TSA-1 TSB TSB-1
1. RBA 8A GTTTCGCTCC 19 - 1 1 5 1 -
2. RBA 13A GTGGATCCGA 21 2 3 2 - 4 -
3. RBA 20A GCGATCCCCA 15 2 3 1 - 4 -
4. RBA 10A CCGCAGCCAA 19 - 2 3 1 3 -
5. RBA 15A AAGAGCCCGT 25 2 2 3 - 1 -
TSA: Treated sample A; TSA-1: Treated sample A-1; TSB: Treated sample B; TSB-1: Treated sample B-1; -, No band
Table 3: DNA polymorphism analyzed by random amplied polymorphic DNA (RAPD) analysis.
Primer C and TSA C and TSA-1 C and TSB C and TSB-1 TSA and TSA-1 TSB and TSB-1 TSA and TSB TSA-1 and TSB-1
%
RBA 8A 15 53 15 7 53 7 0 46
RBA 13A 53 60 100 60 71 68 47 0
RBA 20A 50 35 57 35 22 33 7 0
RBA 10A 54 90 36 54 91 100 18 36
RBA 15A 56 31 25 18 52 62 31 13
Average polymorphism 45 53 46 34 57 54 20 19
C: Control; TSA: Treated Sample A; TSA-1: Treated Sample A-1; TSB: Treated Sample B; TSB-1: Treated Sample B-1
Table 4: Level of polymorphism between control and treated samples.
Alignment View AN Alignment Results Sequence Description
9A1 0.99 Sample studied
9A 0.89 Enterobacter aerogenes
AB244467 0.99 Enterobacter aerogenes strain: C1111
AB244456 0.98 Enterobacter aerogenes strain: An19-2
AJ251468 0.98 Enterobacter aerogenes strain NCTC10006T
AM184245 0.97 Kluyvera cryocrescens strain WAB1904
AJ251467 0.98 Klebsiella ornithinolytica strain JCM6096T
X93216 0.98 Klebsiella planticola strain DR3
AB364958 0.98 Raoultella ornithinolytica
AB353045 0.98 Klebsiella oxytoca strain: No.5
AB094655 0.98 Nocardia beijingensis strain: IFM 10052
AB094654 0.98 Nocardia beijingensis strain: IFM 0915
AN: GenBank Accession Number
Table 5: The closest sequences of Nocardia otitidis from sequence alignment using NCBI GenBank and ribosomal database project (RDP).
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al. (2015) Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of
Bioeld Treated Nocardia otitidis. Biol Syst Open Access 4: 143. doi:10.4172/2329-6577.1000143
Page 5 of 6
Volume 4 • Issue 2 • 1000143
Biol Syst Open Access
ISSN: 2329-6577 BSO, an open access journal
followed by 16S gene sequencing, the coded sample (9A) was detected
as Enterobacter aerogenes which was closely related to the Kluyvera
cryocrescens with 98% similarity and the lowest genetic distance were
0.018 base substitutions per site. Based on above ndings, authors
assumed the sustainability of e Trivedi Eect® in subcultured sample
(9A1), as similar type of data was observed in the case of sub-cultured
sample of N. otitidis.
Due to microbial resistance to a single drug or multiple drugs, the
invention of an eective antimicrobial therapy for the human-wellness
is urgently required. However, due to some limitations of science,
the progress of new medications is slow and very challenging for
scientists. Bioeld treatment might be responsible for doing alteration
in microorganism at the genetic level and/or enzymatic level, which
may act on the receptor protein. While altering the receptor protein,
ligand-receptor/protein interactions may altered that could lead to
show dierent phenotypic characteristics [38]. Based on these results,
it is expected that bioeld treatment has the scope to be a cost eective
and alternative approach than the existing antimicrobial therapy in
near future (Table 5 and 6).
Conclusions
Altogether, the bioeld treatment has altered the MIC values
(16.67%) of tested antimicrobials against the strain of N. otitidis. Using
RAPD markers, the sample was characterized and showed interspecic
relationships with N. otitidis aer bioeld treatment. e molecular
method using 16S rDNA analysis showed that samples were detected as
Enterobacter aerogenes (GenBank Accession Number: AJ251468) with
98% identity of gene sequencing data that was nearest homolog species
to Kluyvera cryocrescens (Accession No. AM184245). e results suggest
that there is an impact of bioeld treatment on MIC, 16S rDNA analysis,
and DNA polymorphism of N. otitidis. ese changes were found in the
organism may be due to alterations happened at the genetic level aer
bioeld treatment. Overall, it seems that Mr. Trivedi’s unique bioeld
energy treatment might be used as an alternate treatment approach in
future antimicrobial therapy.
Acknowledgement
Authors gratefully acknowledged to Trivedi science, Trivedi testimonials and
Trivedi master wellness and the whole team of PD Hinduja National Hospital
and MRC, Mumbai, Microbiology Lab for their support. Authors also would like to
thanks Bangalore Genei Pvt. Ltd., for conducting 16S rDNA sequencing and RAPD
analysis.
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Distance Matrix
AN 1 2 3 4 5 6 7 8 9 10 11 12
AB244456 1 0.992 1 0.990 1 0.992 0.982 0.988 0.720 0.720 0.993 1
AJ251467 2 0.008 0.992 0.998 0.992 1 0.973 0.991 0.720 0.720 0.991 0.992
AB244467 3 0.000 0.008 0.990 1 0.992 0.982 0.988 0.720 0.720 0.993 1
X93216 4 0.010 0.002 0.010 0.990 0.998 0.971 0.988 0.719 0.719 0.988 0.990
AJ251468 5 0.000 0.008 0.000 0.010 0.992 0.982 0.988 0.720 0.720 0.993 1
AB364958 6 0.008 0.000 0.008 0.002 0.008 0.973 0.991 0.720 0.720 0.991 0.992
9A 7 0.018 0.027 0.018 0.029 0.018 0.027 0.969 0.702 0.702 0.974 0.982
AB353045 8 0.012 0.009 0.012 0.012 0.012 0.009 0.031 0.721 0.721 0.989 0.988
AB094654 9 0.280 0.280 0.280 0.281 0.280 0.280 0.298 0.279 1 0.717 0.720
AB094655 10 0.280 0.280 0.280 0.281 0.280 0.280 0.298 0.279 0.000 0.717 0.720
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9A1 12 0.000 0.008 0.000 0.010 0.000 0.008 0.018 0.012 0.280 0.280 0.007
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(above diagonal) and distance (below diagonal) identities between the studied sample ‘9A’ and ten closest homologs microbe.
Figure 2: Phylogenetic tree of the partial 16S rDNA gene sequencing of
Nocardia otitidis using MEGA 3.1 software using neighbor joining method
(Numbers represent GenBank accession number).
Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al. (2015) Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of
Bioeld Treated Nocardia otitidis. Biol Syst Open Access 4: 143. doi:10.4172/2329-6577.1000143
Page 6 of 6
Volume 4 • Issue 2 • 1000143
Biol Syst Open Access
ISSN: 2329-6577 BSO, an open access journal
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Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, et al. (2015)
Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of Bioeld
Treated Nocardia otitidis. Biol Syst Open Access 4: 143. doi:10.4172/2329-
6577.1000143
... Biofield Energy Healing Treatment has gained rapid rapport as a holistic alternative and complementary medicine therapy that has significant impact on living organisms and nonliving materials without any adverse effects and in a manner that is more cost-effective than more conventional methods. Biofield Energy Treatment (The Trivedi Effect ® ) results has been published in numerous peer-reviewed science journals with significant outcomes in many scientific fields such as cancer research [15], microbiology [16][17][18], biotechnology [19], genetics [20,21], pharmaceutics [22,23], nutraceuticals [24], organic compounds [25,26], agricultural science [27][28][29], and altered the structure of the atom in many metals, ceramics, polymers and chemicals in materials science [30][31][32]. In this study, the authors sought to explore the impact of the Biofield Energy Treatment (The Trivedi Effect ® ) on the given herbomineral formulation and Biofield Energy Treatment per se to the animals, which might improve the immunomodulatory function with respect to the antibody titre, delayed type hypersensitivity reaction, body weight change, feed consumption, hematological parameters, and serum biochemistry using standard assays. ...
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A new proprietary herbomineral formulation was formulated, consisting of an essential ingredients viz. an herbal root extract ashwagandha and minerals (zinc, magnesium, and selenium). The aim of this study was to evaluate the immunomodulatory potential of Energy of Consciousness Healing (The Trivedi Effect®) Treatment on the herbomineral formulation in male Sprague Dawley rats. The test formulation was divided into two parts. One part was denoted as the control without any Biofield Energy Treatment, while the other part was defined as the Biofield Energy Treated sample, which received the Biofield Energy Healing Treatment remotely from seven renowned Biofield Energy Healers. Additionally, one group of animals was also received Biofield Energy Treatment per se (day -15) by Biofield Energy Healers under similar conditions. The immunological parameters viz. humoral immune analysis, paw volume, hematological study, biochemistry, body weight, feed and water intake, and histopathology analysis were performed in this experiment. The humoral immune response data showed the secondary antibody titre was significantly increased by 112.50% and 87.50% in the Biofield Energy Treated test formulation group (G4) and untreated test formulation group (G5) compared to the disease control group (G2). The results of delayed type hypersensitivity showed significant increased the paw volume by 111.76%, 51.45%, 100%, and 64.80% in the G4, G5, Biofield Energy Treatment group per se at day -15 (G6) and Biofield Energy Treated test formulation at day -15 (G7) groups, respectively compared to the G2 group. The platelet count was significantly increased by 37.72%, 8.69%, 16.30% and 33.11% in the G4, G5, G6, and G7 groups, respectively compared to the G2 group. The level of blood urea nitrogen was significantly decreased by 14.01% in the G4 group compared to the disease control. Moreover, the concentration of uric acid was significantly reduced by 28.97% in the G7 group compared to the G2 group. Animal weight parameters suggested that there were no treatment-related changes in any group. Organ to body weight ratio, feed and water intake data described that the Biofield Energy Treated test formulation was found to be safe without any side-effect during the course of the experiment. Overall, results suggested that the Biofield Energy Treated herbomineral formulation and Biofield Energy Treatment per se can be used for autoimmune and inflammatory diseases, stress management and prevention, and anti-aging by improving overall health.
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... This energy can be harnessed and transmitted by individuals into living and non-living things via the process of Biofield Energy Healing. Biofield Energy Treatment (The Trivedi Effect ® ) has been extensively studied with significant outcomes in many scientific fields such as cancer science [16, 17], altering microbial characteristics and features including changing the microbial sensitivity of pathogenic microbes in microbiology [18][19][20][21], genetics [22, 23], altered physical and chemical compounds in pharmaceutics [24][25][26][27], improved the overall productivity, quality and yield of crops and plants in agricultural science [28][29][30][31], and in materials science where The Trivedi Effect ® has demonstrated its ability to alter the structural, thermal and physical properties of metals, polymers, ceramics and chemicals [32][33][34][35]. The authors of this study sought to evaluate the impact of Biofield Energy Treatment (The Trivedi Effect ® ) on the given herbomineral formulation, which might improve the immunomodulatory function in an in vitro cellular model of mice splenocyte cells. ...
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... This energy can be harnessed and transmitted by individuals into living organisms and non-living things via the process of Biofield Energy Healing. Biofield Energy Treatments (The Trivedi Effect ® ) have been extensively studied with significant outcomes in many scientific fields such as cancer science [16, 17], altering microbial characteristics and features including changing the microbial sensitivity of pathogenic microbes in microbiology [18][19][20][21], genetics [22, 23], altered physical and chemical compounds in pharmaceutics [24][25][26][27], improved the overall productivity, quality and yield of crops and plants in agricultural science [28][29][30][31], and in materials science where The Trivedi Effect ® has demonstrated its ability to alter the structural, thermal and physical properties of metals, polymers, chemicals and ceramics [32][33][34][35]. The authors of this study sought to evaluate the impact of Biofield Energy Treatment (The Trivedi Effect ® ) on the given herbomineral formulation, which might improve its immunomodulatory function in an in vitro cellular model with mice splenocyte cells. ...
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... This energy can be harnessed and transmitted by individuals into living and non-living things via the process of Biofield Energy Healing. Biofield Energy Treatment (The Trivedi Effect ® ) has been extensively studied with significant outcomes in many scientific fields such as cancer science [16, 17], altering microbial characteristics and features including changing the microbial sensitivity of pathogenic microbes in microbiology [18][19][20][21], genetics [22, 23], altered physical and chemical compounds in pharmaceutics [24][25][26][27], improved the overall productivity, quality and yield of crops and plants in agricultural science [28][29][30][31], and in materials science where The Trivedi Effect ® has demonstrated its ability to alter the structural, thermal and physical properties of metals, polymers, chemicals and ceramics [32][33][34][35]. The authors of this study sought to evaluate the impact of Biofield Energy Treatment (The Trivedi Effect ® ) on the given herbomineral formulation, which might improve the immunomodulatory function in in vitro cellular model on mice splenocyte cells. ...
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Due to the increased popularity of herbomineral preparations in the healthcare sector, a new proprietary herbomineral formulation was formulated consisting of ashwagandha root extract and three minerals viz. zinc chloride, magnesium gluconate, and sodium selenate. The objective of the study was to evaluate the in vitro effect of Biofield Energy Healing (The Trivedi Effect®) on the test formulation using murine splenocyte cells. The herbomineral formulation was divided into two parts; one defined as the control, while the other part was treated with the Biofield Energy Healing Treatment performed from a remote distance by twenty renowned Biofield Energy Healers (The Trivedi Effect®) and defined as the Biofield Treated formulation. The splenocyte cells were exposed to test formulations at concentration from 0.00001053 to 10.53 µg/mL and were analyzed after 48 hours for cell viability using MTT assay. The expression of the cytokines (TNF-α, IFN-γ, IL-1β, and MIP-1α) was determined using ELISA assay. The cell viability data showed that all the tested concentration ranges were found to be safe with percentage cell viability at more than 80%. Further, TNF-α expression was significantly inhibited in the Biofield Treated test formulation group with respect to the vehicle control, while at 0.001053 and 0.1053 µg/mL, the expression was suppressed by 1.70% and 8.16%, respectively in the Biofield Treated test formulation compared to the untreated formulation. However, a significant immunosuppression was reported in IFN-γ expression at 0.00001053, 0.0001053, 0.01053, 0.1053, and 1.053 µg/mL by 12.63%, 2.31%, 8.31%, 9.15%, and 7.86%, respectively in the Biofield Treated test formulation compared with the untreated test formulation. The MIP-1α expression was inhibited by 8.31%, 21.53%, and 8.70% at 0.0001053, 0.01053, and 0.1053 µg/mL, respectively in the Biofield Treated formulation compared with the untreated test formulation. However, IL-1β expression was significantly suppressed by 19.72% at concentration 0.00001053 µg/mL in the Biofield Treated test formulation compared with the untreated test formulation. Thus, the down-regulation of tested cytokines and chemokines in the Biofield Energy Healing test formulation might be applicable for controlling acute and chronic inflammation in many clinical diseases. Overall, the results demonstrated that The Trivedi Effect®- Biofield Energy Healing (TEBEH) has the capacity to potentiate the immunomodulatory activity of the test formulation, which can be useful against autoimmune disorders. Biofield Treated Test formulation may also be useful in anti-aging, anti-inflammatory, stress management and in preventing immune-mediated tissue damage in organ transplants by improving overall health and quality of life.
... This energy can be harnessed and transmitted by individuals into living and non-living things via the process of Biofield Energy Healing. Biofield Energy Treatment (The Trivedi Effect ® ) has been extensively studied with significant outcomes in many scientific fields such as cancer research [16, 17], altered antimicrobial sensitivity of pathogenic microbes in microbiology [18][19][20][21], genetics [22, 23], altered physical and chemical properties of pharmaceutical compounds [24][25][26][27], improved overall growth and yield of plants in agricultural science [28][29][30][31], and in changing the structure of the atom in relation to various metals, ceramics, polymers and chemicals in materials science [32][33][34][35]. The authors of this study want to evaluate the impact of Biofield Energy Treatment (The Trivedi Effect ® ) on the given herbomineral formulation, which might improve the immunomodulatory function in in vitro cellular models on mice splenocyte cells. ...
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The use of herbomineral formulations in the healthcare sector has increased due to their high safety and better therapeutic action. A new proprietary herbomineral formulation was formulated with a mixture of the herbal root extract of ashwagandha and three minerals viz. zinc chloride, magnesium gluconate, and sodium selenate. The aim of the study was to evaluate the immunomodulatory potential of Biofield Energy Healing (The Trivedi Effect®) on the formulation when applied to splenocyte cells isolated from mice spleen. The formulation was divided into two parts; one was the control without any Biofield Energy Treatment, while the other part was defined as the Biofield Energy Treated sample, which received Biofield Energy Healing Treatment remotely by seven renowned Biofield Energy Healers. The test formulation was evaluated to find the expression of pro-inflammatory cytokines such as TNF-α, MIP-1α, and IL-1β along with non-cytotoxic concentrations by MTT assay. The splenocytes were given the Biofield Energy Treated and untreated sample at concentrations range (0.00001053 to 10.53 µg/mL) for 48 hours and was reported with safe concentration up to 1.053 µg/mL with percentage viability range from 76.7% to 109.2% in both samples. Biofield Energy Healing significantly enhanced the cell viability as compared with the untreated formulation. The expression of TNF-α was significantly inhibited in the Biofield Treated formulation at 0.01053, 0.1053, and 1.053 µg/mL by 1.77%, 1.93%, and 3.55%, respectively compared with the untreated formulation. The rest of the tested concentrations of the Biofield Treated formulation showed an increase in TNF-α expression at 0.00001053, 0.0001053, and 0.001053 µg/mL by 7.26%, 8.50%, and 8.50%, respectively compared to the vehicle control group. Similarly, the MIP-1α expression was inhibited by the Biofield Energy Treated formulation and showed immunosuppression activity at 0.01053 µg/mL by 18.47% (p≤0.001) compared to the untreated formulation. MIP-1α expression was reported as 628.94 ± 13.0 pg/mL in the untreated formulation, while it decreased to 512.74 ± 1.9 pg/mL in the Biofield Treated formulation at 0.01053 µg/mL. In addition, the IL-1β secretion was also significantly inhibited by the Biofield Treated formulation at concentrations 0.001053, 0.01053, 0.1053, and 1.053 µg/mL by 72.02%, 50.16%, 30.68%, and 22.11%, respectively as compared with the untreated formulation. Overall, The Trivedi Effect® significantly down-regulated the pro-inflammatory cytokines and potentiated the immunosuppressive effect of the treated formulation, which can be better utilized in organ transplants, anti-aging, stress management, autoimmune disorders, and inflammatory disorders, etc. to modulate the immune system by improving overall health.
... The energy can be harnessed from the universe and then, it can be applied to the living and non-living objects to achieve the alterations in the characteristic properties by the healing practitioner. The applications of The Trivedi Effect ® have gained scientific attention in the field of chemical science [17][18][19][20] 24], materials science [25][26][27], agricultural science [28][29][30]genetics [31][32][33][34], biotechnology [35][36], nutraceuticals [37] pharmaceuticals [38][39][40], and medical sciences [41, 42]. The mass spectrometry (MS) technique is the main choice, and the conventional analytical technique gas chromatography-mass spectrometry (GC-MS) can perform isotope ratio measurement at low micro molar concentration levels with sufficient precision [43][44][45][46]. ...
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... The energy from the universe can be harnessed by the expert, and it can be applied to the living and non-living objects to achieve the alterations in the characteristic properties. Various applications of The Trivedi Effect ® have achieved a milestone and recognized scientifically in the field of chemical science [17][18][19][20] 24], materials science [25][26][27], agricultural science [28][29][30], biotechnology [31, 32], genetics [33, 34], nutraceuticals [35][36][37], pharmaceuticals [38][39][40], and medical sciences [41, 42]. The mass spectrometry (MS) technique is a choice for the isotope ratio analysis [43]. ...
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