slide 1: Biological Systems: Open Access
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 Trivedi
1
Alice Branton
1
Dahryn Trivedi
1
Gopal Nayak
1
Sambhu Charan Mondal
2
and Snehasis Jana
2
1
Trivedi Global Inc. 10624 S Eastern Avenue Suite A-969 Henderson NV 89052 USA
2
Trivedi 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 biofeld energy treatment on N. otitidis and analyzed for antimicrobial susceptibility
pattern minimum inhibitory concentration MIC DNA polymorphism by Random Amplifed 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 biofeld 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 biofeld energy treatment has a signifcant 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:
publicationtrivedisrl.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 Biofeld 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 1000 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 ofen used as the
drug of choice for the treatment of nocardiosis 12. Terefore some
alternative strategies are needed to treat against nocardiosis. Biofeld
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 biofeld putative energy
felds 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 signifcance 14. Recently it was
discovered that all the electrical processes happening in the human
body have strong relationship with the magnetic feld as required by
Keywords: Nocardia otitidis Nocardiosis Antimicrobial
susceptibility Biofeld energy treatment 16S rDNA sequencing
Random amplifed 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 Amplifed Polymorphic DNA CNS:
Central Nervous System
Introduction
Te 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
difer from the mycobacteria by the possession of short-chain 40 to
60 carbons mycolic acids 1. Nocardia otitidis N. otitidis is a weak
flamentous Gram-positive catalase-positive branching rods shaped
bacterium that appears similar to Actinomyces species. However it
can usually be diferentiated from Actinomyces by acid-fast staining
2. Te 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. Te 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 fnd
any diference between two genera 4. Based on immunoblot and
enzyme-linked immunosorbent assay ELISA techniques that detect
specifc antibodies that appear as common in various Nocardia and
Actinomadura species. Tese 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. Te most common manifestation of Nocardial disease is
pulmonary nocardiosis 7 extrapulmonary disease 8 and ocular
slide 2: Citation: Trivedi MK Branton A Trivedi D Nayak G Mondal SC et al. 2015 Evaluation of Antibiogram Genotype and Phylogenetic Analysis of
Biofeld 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
felds in the surrounding space 1516. Tus 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 biofeld. Terefore the biofeld consists of an
electromagnetic feld being generated by moving electrically charged
particles ions cell molecule etc. inside the human body 17. Te
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 scientifc instruments such as Kirlian photography polycontrast
interference photography PIP and resonance feld imaging RFI can
be extensively used to measure the biofeld of human body 19. Tus
human has the ability to harness the energy from the environment or
universe and can transmit into any living or nonliving objects around
the Globe. Te objects always receive the energy and responding into
the useful way that is called biofeld energy and the process is known as
biofeld energy treatment. Mr. Mahendra T rivedi’ s unique biofeld energy
treatment Te Trivedi Efect® has been known to improve the overall
productivity of crops 2021 altered characteristics features of microbes
22-24 alter the structural physical and thermal properties of several
metals 2526 and improved growth and anatomical characteristics of
various medicinal plants 2728.
Based on clinical signifcance of N. otitidis and signifcant impact
of Mr. Trivedi’s biofeld energy modality on microbes the present work
was undertaken to evaluate the impact of Mr. Trivedi’s biofeld energy
on N. otitidis in relation to antimicrobials susceptibility minimum
inhibitory concentration MIC random amplifed 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 diferent sealed packs were stored with proper storage
conditions until further use. All the tested antimicrobials were procured
from Sigma-Aldrich India. Te 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. Te 16S rDNA sequencing and DNA
fngerprinting RAPD studies were carried out using Ultrapure
Genomic DNA Prep Kit Cat KT 83 Bangalore Genei India.
Experimental design and biofeld treatment strategy
N. otitidis strain was divided into two groups i.e. control and
treated. Te treated group was in sealed pack and handed over to Mr.
Trivedi for biofeld energy treatment under laboratory conditions. Mr.
Trivedi provided the treatment through his energy transmission process
to the treated group without touching the sample. Afer 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. Tese two biofeld 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. Te result of treated sample was compared with respect to
the control. Te 16S rDNA analysis was performed on biofeld treated
samples A and its subcultured sample A1.
Antimicrobial susceptibility test
Te antimicrobial susceptibility testing is important for clinically
signifcant species. Te 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. Te detailed
experimental procedure and conditions were followed as per the
manufacturer’s instructions. Te 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 amplifed polymorphic DNA RAPD analysis
For DNA fngerprinting 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 fve
RAPD primers which were labelled as RBA8A RBA13A RBA20A
RBA10A and RBA15A were adopted from earlier studies. Te PCR
mixture contained 2.5 μL each of bufer 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 MgCl
2
and 9.5 μL of water in
a total of 25 μL with the following PCR amplifcation 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 fnal extension
at 72°C for 7 min. Amplifed 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. Te
percentage of polymorphism was calculated using following equation:
Percent polymorphismA/B × 100
Where ANumber of polymorphic bands in treated sample and
BNumber of polymorphic bands in control.
Amplifcation and gene sequencing of 16S rDNA
Genomic DNA was isolated from N. otitidis cells set A sample coded
as 9A using genomic purifcation kit according to the manufacturer
instructions. 16S rDNA gene 1.5 kb fragment was amplifed with the
help of high-fdelity polymerase chain reaction PCR using universal
primers forward primer 5 ’-AGAGTTTGATCCTGGCTCAG-3 ’ and
reverse primer 3’-ACGGTCATACCTTGTTACGACTT-5 ’. Amplifed
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. Te PCR amplifed
fragment was purifed from the agarose gel using a DNA gel extraction
kit. Sequencing of amplifed product was done on a commercial basis
from Bangalore Genei India. Te 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 sofware
31.
Results and Discussion
Antimicrobial susceptibility test
Antimicrobial susceptibility pattern of Nocardia can vary from
slide 3: Citation: Trivedi MK Branton A Trivedi D Nayak G Mondal SC et al. 2015 Evaluation of Antibiogram Genotype and Phylogenetic Analysis of
Biofeld 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. Te therapeutic efectiveness depends on the proper
identifcation of species in infected patients and on in vitro sensitivity
studies 32. Te 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. Te outcome of N. otitidis susceptibility pattern and MIC values of
tested antimicrobials afer biofeld energy treatment are summarized in
Table 1 and 2 respectively. Te 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. Te
treated cells of N. otitidis did not show any alteration with respect to
antimicrobial susceptibility pattern as compared to the control Table
1. Te susceptible nature of N. otitidis to amikacin sulfamethoxazole
and ciprofoxacin and resistance pattern to cefriaxone 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 biofeld 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 afer the biofeld 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 1112. 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 afer Mr. Trivedi’s biofeld energy treatment.
Hence authors assumed that this improvement of MIC value without
alteration of sensitivity pattern may be due to the efect of putative
energy transmit through biofeld healing Table 2.
Random amplifed polymorphic DNA RAPD analysis
Te treated and control samples were identifed on the basis of
their diferent and discriminative RAPD patterns. RAPD is a preferred
tool that is being used now days to correlate the genetic similarity
or mutations between species. Te 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 efciently detected using
the PCR primers and identify inter-strain variations among species in
treated samples 34. Te degree of relatedness and genetic mapping
can be correlated between similar or diferent treated sample species
35.
Te DNA fngerprinting by RAPD analysis using fve primers was
carried out on the control and treated samples. DNA fngerprinting by
RAPD analysis of the control and treated samples are shown in Figure 1
and the polymorphic bands are marked by arrows. Te RAPD patterns
of treated samples showed some unique and dissimilar patterns. DNA
polymorphism analyzed by the RAPD analysis was presented in T able 3.
Te level of polymorphism between control and treated samples A A1
B and B1 are summarized in Table 4. Te level of polymorphism was
found in an average range of 34 to 53 in treated samples as compared
to control in N. otitidis afer the biofeld treatment. Te 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
Te bacteria that are poorly diferentiated by conventional methods
needs molecular analysis method like 16S rDNA sequence 36. Tis
molecular-based technique is a suitable tool for identifcation of most
1000
900
800
700
600
500
400
300
200
100
bp
Figure 1: Random amplifed polymorphic-DNA fragment patterns of Nocardia
otitidis generated using fve 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. Ciprofoxacin S S
10. Gatifoxacin S S
11. Amoxicillin/k-clavulanate R R
12. Trimethoprim/sulfamethoxazole S S
R: Resistant S: Susceptible Control: ATCC strain of N. otitidis without biofeld
energy treatment Treatment: ATCC strain of N. otitidis with Mr. Trivedi’s biofeld
energy treatment
Table 1: Antimicrobial susceptibility pattern of antimicrobials against ATCC strain of
Nocardia otitidis after biofeld 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. Ciprofoxacin 4.0 4.0
10. Gatifoxacin 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 biofeld energy treatment
Treatment: ATCC strain of N. otitidis with Mr. Trivedi’s biofeld energy treatment
Table 2: Minimum inhibitory concentration MIC of antimicrobials against ATCC
strain of Nocardia otitidis after biofeld energy treatment on day 10 as per CLSI
guidelines.
slide 4: Citation: Trivedi MK Branton A Trivedi D Nayak G Mondal SC et al. 2015 Evaluation of Antibiogram Genotype and Phylogenetic Analysis of
Biofeld 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-specifc variability
37. Te 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. Te 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. Te 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 T able 5. Te distance
matrix based on nucleotide sequence homology data are presented in
Table 6. Te phylogenetic tree was established using BLAST-Webpage
NCBI. According to Table 6 ten di ferent related bacterial species
of N. otitidis were selected as Operational Taxonomic Units OTUs
in order to investigate the phylogenetic relationship of N. otitidis.
Tere were 1462 base-pair nucleotides of 16S rDNA gene sequences
which were analyzed and multiple alignments were constructed using
ClustalW in MEGA3.1 31. Te 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.
Tis 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. Te 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. Te results suggested that afer biofeld 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 amplifed 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.
slide 5: Citation: Trivedi MK Branton A Trivedi D Nayak G Mondal SC et al. 2015 Evaluation of Antibiogram Genotype and Phylogenetic Analysis of
Biofeld 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 sequ encing 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 fndings authors
assumed the sustainability of Te Trivedi Efect® 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 efective 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. Biofeld 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 diferent phenotypic characteristics 38. Based on these results
it is expected that biofeld treatment has the scope to be a cost efective
and alternative approach than the existing antimicrobial therapy in
near future Table 5 and 6.
Conclusions
Altogether the biofeld 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 interspecifc
relationships with N. otitidis afer biofeld treatment. Te 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. Te results suggest
that there is an impact of biofeld treatment on MIC 16S rDNA analysis
and DNA polymorphism of N. otitidis. Tese changes were found in the
organism may be due to alterations happened at the genetic level afer
biofeld treatment. Overall it seems that Mr. Trivedi’s unique biofeld
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
AM184245 11 0.007 0.009 0.007 0.012 0.007 0.009 0.026 0.011 0.283 0.283 — 0.993
9A1 12 0.000 0.008 0.000 0.010 0.000 0.008 0.018 0.012 0.280 0.280 0.007 —
AN: GenBank Accession Number
Table 6: Distance matrix Nocardia otitidis samples 9A and 9A1 based on nucleotide sequence homology Using Kimura-2 Parameter indicates nucleotide similarity
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.
slide 6: Citation: Trivedi MK Branton A Trivedi D Nayak G Mondal SC et al. 2015 Evaluation of Antibiogram Genotype and Phylogenetic Analysis of
Biofeld Treated Nocardia otitidis. Biol Syst Open Access 4: 143. doi:10.4172/2329-6577.1000143
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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 Biofeld
Treated Nocardia otitidis. Biol Syst Open Access 4: 143. doi:10.4172/2329-
6577.1000143