Biofield Treated Nocardia otitidis

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The genus Nocardia is associated with the group of microorganisms known as the aerobic actinomycetes and belongs to the family of Mycobacteriaceae.

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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

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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 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

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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 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.

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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 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.

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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 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. References 1. Saubolle MA Sussland D 2003 Nocardiosis: Review of clinical and laboratory experience. J Clin Microbiol 41: 4497-4501. 2. Wilson JW 2012 Nocardiosis: Updates and clinical overview. Mayo Clin Proc 87: 403-407. 3. Brown-Elliott BA Brown JM Conville PS Wallace RJ Jr 2006 Clinical and laboratory features of the Nocardia spp. based on current molecular taxonomy. Clin Microbiol Rev 19: 259-282. 4. Mordarska H Mordarski M Goodfellow M 1972 Chemotaxonomic characters and classifcation of some nocardioform bacteria. J Gen Microbiol 71: 77-86. 5. Angeles AM Sugar AM 1987 Rapid diagnosis of nocardiosis with an enzyme immunoassay. J Infect Dis 155: 292-296. 6. Saubolle AM 2002 Aerobic actinomycetes. Diagnostic microbiology of the immunocompromised host ASM Press Washington DC. 7. Long PF 1994 A retrospective study of Nocardia infections associated with the acquired immune defciency syndrome AIDS Infection 22: 362-364. 8. Filice GA 2005 Nocardiosis in persons with human immunodefciency virus infection transplant recipients and large geographically defned populations. J Lab Clin Med 145: 156-162. 9. Climenhaga DB Tokarewicz AC Willis NR 1984 Nocardia keratitis. Can J Ophthalmol 19: 284-286. 10. Beaman BL Burnside J Edwards B Causey W 1976 Nocardial infections in the United States 1972-1974. J Infect Dis 134: 286-289. 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.

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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 Page 6 of 6 Volume 4 • Issue 2 • 1000143 Biol Syst Open Access ISSN: 2329-6577 BSO an open access journal 11. Benbow EP Jr. Smith DT Grimson KS 1944 Sulfonamide therapy in actinomycosis: Two cases caused by aerobic partially acid-fast Actinomyces. Am Rev Tuberc 49: 395-407. 12. Wallace RJ Jr Septimus EJ Williams TW Jr Conklin RH Satterwhite TK et al. 1982 Use of trimethoprim-sulfamethoxazole for treatment of infections due to Nocardia. Rev Infect Dis 4: 315-325. 13. Koithan M 2009 Introducing Complementary and Alternative Therapies. J Nurse Pract 5: 18-20. 14. Burr HS 1957 Bibliography of Harold Saxton Burr. Yale J Biol Med 30: 163- 167. 15. Hammerschlag R Jain S Baldwin AL Gronowicz G Lutgendorf SK et al. 2012 Biofeld research: A roundtable discussion of scientifc and methodological issues. J Altern Complement Med 18: 1081-1086. 16. Movaffaghi Z Farsi M 2009 Biofeld therapies: Biophysical basis and biological regulations Complement Ther Clin Pract 15: 35-37. 17. Niggli HJ Tudisco S Privitera G Applegate LA Scordino A et al. 2005 Laser- ultraviolet-A-induced ultraweak photon emission in mammalian cells. J Biomed Opt 10: 024006. 18. Sun Y Wang C Dai J 2010 Biophotons as neural communication signals demonstrated by in situ biophoton autography. Photochem Photobiol Sci 9: 315-322. 19. Prakash S Chowdhury AR Gupta A 2015 Monitoring the human health by measuring the biofeld “aura”: An overview. IJAER 10: 27637-27641. 20. Sances F Flora E Patil S Spence A Shinde V 2013 Impact of biofeld treatment on ginseng and organic blueberry yield. Agrivita J Agric Sci 35: 22-29. 21. Lenssen AW 2013 Biofeld and fungicide seed treatment infuences on soybean productivity seed quality and weed community. Agricultural Journal 83: 138-143. 22. Trivedi MK Patil S Shettigar H Bairwa K Jana S 2015 Phenotypic and biotypic characterization of Klebsiella oxytoca: An impact of biofeld treatment. J Microb Biochem Technol 7: 203-206. 23. Trivedi MK Patil S Shettigar H Gangwar M Jana S 2015 An effect of biofeld treatment on multidrug-resistant Burkholderia cepacia: A multihost pathogen. J Trop Dis 3: 167. 24. Trivedi MK Patil S Shettigar H Gangwar M Jana S 2015 Antimicrobial sensitivity pattern of Pseudomonas fuorescens after biofeld treatment. J Infect Dis Ther 3: 222. 25. Dabhade VV Tallapragada RR Trivedi MK 2009 Effect of external energy on atomic crystalline and powder characteristics of antimony and bismuth powders. Bull Mater Sci 32: 471-479. 26. Trivedi MK Nayak G Patil S Tallapragada RM Latiyal O 2015 Studies of the atomic and crystalline characteristics of ceramic oxide nano powders after bio feld treatment. Ind Eng Manage 4: 161. 27. Patil SA Nayak GB Barve SS Tembe RP Khan RR 2012 Impact of biofeld treatment on growth and anatomical characteristics of Pogostemon cablin Benth.. Biotechnology 11: 154-162. 28. Nayak G Altekar N 2015 Effect of biofeld treatment on plant growth and adaptation. J Environ Health Sci 1: 1-9. 29. Woods GL Brown-Elliott BA Desmond EP Hall GS Heifets L et al. 2003 Susceptibility testing of Mycobacteria Nocardiae and other aerobic Actinomycetes Approved Standard. NCCLS document M24-A 2003. 30. Welsh J McClelland M 1990 Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18: 7213-7218. 31. Kumar S Tamura K Nei M 2004 MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence alignment. Brief Bioinform 5: 150-163. 32. Burgert S 1998 Nocardiosis: A clinical review. Infect Dis Clin Pract 8: 27-32. 33. Williams JG Kubelik AR Livak KJ Rafalski JA Tingey SV 1990 DNA polymorphisms amplifed by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18: 6531-6535. 34. Bingen E Boissinot C Desjardins P Cave H Brahimi N et al. 1993 Arbitrarily primed polymerase chain reaction provides rapid differentiation of Proteus mirabilis isolates from a pediatric hospital. J Clin Microbiol 31: 1055-1059. 35. Williams JG Hanafey MK Rafalski JA Tingey SV 1993 Genetic analysis using random amplifed polymorphic DNA markers. Methods Enzymol 218: 704-740. 36. Drancourt M Bollet C Carlioz A Martelin R Gayral JP et al. 2000 16S ribosomal DNA sequence analysis of a large collection of environmental and clinical unidentifable bacterial isolates. J Clin Microbiol 38: 3623-3630. 37. Vandamme P Pot B Gillis M de Vos P Kersters K et al. 1996 Polyphasic taxonomy a consensus approach to bacterial systematics. Microbiol Rev 60: 407-438. 38. Lindstrom E Mild KH Lundgren E 1998 Analysis of the T cell activation signaling pathway during ELF magnetic feld exposure p56 lck and Ca 2+ i -measurements. Bioeletrochem Bioenerg 46: 129-137. OMICS International: Publication Benefits Features Unique features: • Increased global visibility of ar ticles thr ough w or ld wide distribution and inde xing • Sho w casing recent researc h output in a timely and updated manner • Special issues on the current trends of scientifc researc h Special features: • 700 Open Access Journals • 50000 editorial team • R apid review process • Quality and quick editorial review and publication processing • Indexing at PubMed partial Scopus EBSCO Index Copernicus and Google Scholar etc • Sharing Option: Social Networking Enabled • Authors Reviewers and Editors rewarded with online Scientifc Credits • Better discount for your subsequent articles Submit your manuscript at: http://www.omicsonline.org/submission/ 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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