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		Spectroscopic Characterization of Methyl-2-Naphthyl Ether
	

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		Spectroscopic Characterization of Methyl-2-Naphthyl Ether
	

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| Methyl-2-naphthyl ether is an organic compound and used as the primary moiety for the synthesis of several antimicrobial and anti-inflammatory agents. A test to investigate the impact of biofield energy treatment on the physical, thermal, and spectroscopic characterization of methyl-2-naphthyl ether.

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Mahendra Kumar Trivedi1, Alice Branton1, Dahryn Trivedi1, Gopal Nayak1, Khemraj Bairwa2 and Snehasis Jana2*

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

*Corresponding Author:

Snehasis Jana
Trivedi Science Research Laboratory Pvt. Ltd.
Hall-A, Chinar Mega Mall, Chinar Fortune City
Hoshangabad Rd., Bhopal-462026 Madhya Pradesh India
Tel: 91-755-6660006
E-mail: [email protected]

Received date: September 07, 2015; Accepted date: September 14, 2015; Published date: September 20, 2015

Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Bairwa K, et al. (2015) Physical, Thermal, and Spectroscopic Characterization of Biofield Energy Treated Methyl-2-Naphthyl Ether. J Environ Anal Chem 2:162. doi:10.4172/2380-2391.1000162

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.

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Abstract

Methyl-2-naphthyl ether (MNE) is an organic compound and used as the primary moiety for the synthesis of several antimicrobial and anti-inflammatory agents. This study was attempted to evaluate the impact of biofield energy treatment on the physical, thermal, and spectroscopic properties of MNE. The study was carried out in two groups i.e., control and treated. The treated group was subjected to Mr. Trivedi's biofield treatment. Afterward, the control and treated samples of MNE were evaluated using X-ray diffraction (XRD), surface area analyzer, differential scanning calorimetry (DSC), thermogravimetric analysis-derivative thermogravimetric analysis (TGA-DTG), Fourier transform infrared (FTIR), and ultraviolet-visible (UV-Vis) spectroscopy. The XRD study exhibited the decrease in average crystallite size by 30.70%. The surface area analysis showed 5.32% decrease in surface area of the treated sample with respect to the control. The DSC thermogram of treated MNE exhibited no significant change in the melting temperature; however, the latent heat of fusion was slightly increased (0.83%) after biofield treatment as compared to the control sample. The TGA analysis showed the onset temperature of thermal degradation at 158℃ in the control sample that was reduced to 124℃ after biofield treatment. The result showed about 21.52% decrease in onset temperature of thermal degradation of treated MNE as compared to the control. Similarly, the end-set temperature of thermal degradation was also reduced by 13.51% after biofield treatment with respect to the control. The FT-IR and UV spectroscopic studies did not show any changes in the wavenumber and wavelength, respectively in treated MNE with respect to the control. Overall, the XRD, surface area and thermal analysis suggest that biofield treatment has the impact on physical and thermal properties of the treated MNE as compared to the control.

Keywords

Methyl-2-naphthyl ether; Biofield energy; X-ray diffraction; Surface area analysis; Differential scanning calorimetry; Thermogravimetric analysis

Abbreviations

MNE: Methyl-2-Naphthyl Ether; NCCAM: National Center For Complementary And Alternative Medicine; XRD: X-Ray Diffraction; DSC: Differential Scanning Calorimetry; TGA: Thermogravimetric Analysis; DTA: Differential Thermal Analysis; DTG: Derivative Thermogravimetry; FT-IR: Fourier Transforms Infrared

Introduction

Naphthalene has been described as new class of potent antimicrobials against wide range of human pathogens. It occupies a central place among biologically active compounds owing to its varied and exciting antibiotic properties with less toxicity [ 1]. Numerous naphthalene containing antimicrobial drugs are existing like naftifine, nafacillin, terbinafine, tolnaftate, etc. that plays vital role against microbial infections [ 2, 3]. Further, the naphthalene derivatives like naproxen, nabumetone were also studied in depth as the nonsteroidal anti-inflammatory drugs (NSAIDs). They are mainly nonselective inhibitor of two cyclooxygenase (COX) isoform i.e., COX-1 and COX-2 [ 4, 5]. Moreover, the structurally relative naphthalene derivatives lower the parathyroid level by binding to calcium receptor on the parathyroid gland. Thus, it helps to regulate hyperparathyroidism especially in kidney disease or parathyroid gland neoplasm [ 6]. Based on the importance of naphthalene derivative as a main moiety for organic synthesis of several pharmaceutical drugs, it is advantageous to find out the alternate approach that can enhance the physicochemical and thermal properties of naphthalene derivative i.e., methyl-2-naphthyl ether (MNE). Recently, an alternate treatment approach i.e., healing therapy or therapeutic touch, known as the biofield energy treatment, which was reported in several fields. The National Institute of Health/ National Center for Complementary and Alternative Medicine (NIH/ NCCAM) conceived the biofield energy treatment in subcategory of energy therapies (putative energy fields) [ 7, 8]. The biofield treatment is being used in healing process to reduce pain, anxiety and to promote the overall health of human being [ 9, 10]. Biofield is an electromagnetic field that permeates and surrounds living organisms. This biologically produced electromagnetic and subtle energy field regulates the various physiological and communications functions within the human organism [ 11]. Researchers have attempted different biological studies and effects of on various biomolecules such as proteins, antibiotics [ 12], bacterial cultures [ 13], and conformational change in DNA [ 14]. Prakash et al. reported that various scientific instruments such as Kirlian photography, resonance field imaging (RFI) and polycontrast interference photography (PIP) could be extensively used to measure the biofield of human body [ 15]. Thus, the human has the ability to harness the energy from the environment or Universe and transmit it to any living or nonliving object on the Globe. The object(s) receive the energy and respond into the useful way; this process is termed as biofield treatment. Mr. Trivedi's unique biofield energy treatment is known as The Trivedi Effect®. Recently, biofield energy treatment has been described as an alternative method to alter the physicochemical and thermal properties of several metals and ceramics [ 16- 18]. It has also reported to alter the spectroscopic properties of various pharmaceutical drugs like paracetamol, piroxicam, metronidazole, and tinidazole [ 19, 20]. Moreover, the biofield treatment has been studied in several fields like agriculture research [ 21, 22], biotechnology research [ 23], and microbiology research [ 24, 25].

Based on the published literature and outstanding impact of biofield energy treatment on various living and nonliving things, the present study was aimed to evaluate the impact of Mr. Trivedi's biofield energy treatment on physical, thermal and spectroscopic properties of MNE using several analytical techniques.

Materials and Methods
Study design
The Methyl-2-Naphthyl Ether was procured from Sisco Research Laboratories, India. The study was performed in two groups i.e., control and treated. The control sample was remained as untreated, and the treated group in sealed pack was handed over to Mr. Trivedi for biofield energy treatment under laboratory conditions. Mr. Trivedi provided the biofield energy treatment to the treated group through his unique energy transmission process without touching the sample. Subsequently, the control and treated samples of MNE were analyzed using several analytical techniques like X-ray diffraction (XRD), surface area analysis, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), Fourier transform infrared (FTIR), and ultraviolet-visible (UV-Vis) spectroscopy.
XRD study
The XRD analysis of the control and treated MNE was carried out on Phillips, Holland PW 1710 X-ray diffractometer with nickel filter and copper anode. The wavelength used in XRD system was 1.54056 Å. The XRD diffractograms were obtained in the form of a chart of 2θ vs. intensity. The crystallite size (G) and percent change in crystallite size of MNE were calculated using the following equations [ 26].

Crystallite size (G) = kλ/(bCosθ)

Percent change in crystallite size (G) = [(G t-G c)/G c] ×100

Where, G c and G t are average crystallite size of control and treated powder samples respectively.
Surface area analysis
The surface area of the control and treated MNE was measured using the Brunauer-Emmett-Teller (BET) surface area analyzer, Smart SORB 90. Percent change in surface area was calculated using following equation:

Where, S Control and S Treated are the surface area of the control and treated samples, respectively.
DSC study
The control and treated samples of MNE were studied using a Pyris-6 Perkin Elmer differential scanning calorimeter on a heating rate of 10ºC/min under air atmosphere with air flow rate of 5 mL/ min. An empty pan sealed with cover was used as a reference pan. The melting temperature (T m) and latent heat of fusion (ΔH) were obtained from the DSC curve.
TGA-DTG analysis
TGA-DTG analysis was conducted to investigate the thermal stability of the control and treated MNE. The studies were carried out on Mettler Toledo simultaneous TGA-DTG system. Both the control and treated samples were heated from room temperature to 400ºC with a heating rate of 5ºC/min under air atmosphere. The onset temperature of thermal degradation and percent change in temperature at which maximum weight loss occur in samples were obtained from DTG thermogram.
Spectroscopic studies
For the purpose of FT-IR and UV-Vis spectroscopic characterization, the treated sample was divided into two groups i.e., T1 and T2. Both the treated groups were then analyzed for spectroscopic characteristics employing FT-IR and UV-Vis spectroscopy and data were compared with the respective spectrum of control sample.
FT-IR spectroscopic characterization
The samples for FT-IR spectroscopy were prepared by crushing into fine powder and then mixing in spectroscopic grade KBr. Finally, the mixture was pressed into pellets with a hydraulic press and then used for analysis. The spectra were recorded from Shimadzu's Fourier transform infrared spectrometer (Japan) with the frequency range of 4000-500 cm⁻¹. The analysis was done to evaluate the impact of biofield energy treatment at the atomic levels like force constant, dipole moment, and bond strength in chemical structure [ 27].
Uv-Vis spectroscopic analysis
The samples solutions for UV spectroscopy were prepared in methanol. UV spectra of the control and treated samples of MNE were acquired from Shimadzu UV-2400 PC series spectrophotometer with quartz cuvette having slit widths of 2.0 nm. The wavelength was set between 200-400 nm for analysis. The study was carried out to evaluate the impact of biofield energy treatment on the energy gap of highest occupied molecular orbital and lowest unoccupied molecular orbital (HOMO-LUMO gap) [ 27].

Results and Discussion
XRD analysis
XRD diffractograms of both control and treated MNE are shown in Figure 1. The control sample exhibited the XRD peaks at 2θ equal to 10.27º, 15.44º, 19.07º, 20.57º, 21.21º, 24.76º, 25.33º, and 28.77º. Likewise, the XRD diffractogram of treated MNE showed the XRD peaks at 2θ equal to 10.25º, 15.30º, 19.01º, 20.55º, 21.17º, 24.67º, 25.25º, and 28.84º. The intensity of peak was also altered after biofield energy treatment as compared to the control. XRD diffractograms of both the control and treated MNE showed the intense peaks, which suggest the crystalline nature of MNE. The decrease of XRD peaks intensity in treated sample might be attributed to the reduction in crystallinity and decrease in long-range order of the molecules. The crystallite size was calculated using Scherrer formula and the results are shown in Figure 2. The average crystallite size of the control MNE was observed as 84.55 nm that was decreased up to 58.59 nm in the treated sample. The result indicated about 30.70% decrease in average crystallite size of treated sample with respect to the control.

The increase in internal micro strain leads to decrease the corresponding crystallite size of the material [ 28]. Moreover, Zhang et al. reported that presence of strain and increased atomic displacement from their ideal lattice positions lead to reduction in crystallite size [ 29]. Hence, it is assumed that biofield energy treatment probably induce the internal strain in treated MNE sample. This might be responsible for the decrease in crystallite size of the treated MNE as compared to the control.
Surface area analysis
The surface area of control and treated samples of MNE were determined using BET surface area analyzer and data are presented in Figure 3. The surface area of the control and treated sample was observed as 0.4904 m ²/g and 0.4643 m ²/g, respectively. The result showed a considerable decrease in surface area i.e., 5.32% in the treated MNE as compared to the control sample. It is assumed that biofield energy treatment possibly induced the disappearance of inter-particle boundaries that may lead to the aggregation of particles and thus increase in particle size [ 30]. Presumably, this increase in particle size might lead to decrease in surface area of the treated sample.
DSC analysis
DSC analysis was performed to determine the melting temperature and latent heat of fusion (ΔH) of the control and treated MNE. DSC thermograms ( Figure 4) of MNE showed the melting temperature at 73.72ºC in the control and 73.66ºC in the treated sample ( Table 1). The result depicted no significant change in melting temperature of treated sample as compared to the control. The melting temperature of control MNE was well supported by literature data [ 31]. DSC thermogram exhibited the ΔH of 147.49 J/g in control sample and 148.72 J/g in treated sample of MNE. The result showed about 0.83% increase in latent heat of fusion of treated sample as compared to the control. It is hypothesized that biofield energy treatment may cause absorption of energy during the phase transition from solid to liquid that might lead to increase the latent heat of fusion of treated sample with respect to the control. Previously, our group has reported that biofield energy treatment altered the value of latent heat of fusion in lead and tin powders [ 7].

T max: temperature at maximum weight loss occurs

Table 1: Thermal analysis of control and treated samples of methyl-2-naphthylether.
TGA-DTG analysis
The TGA and DTG thermograms of control and treated samples of MNE are shown in Figure 5 and data are reported in Table 1. The onset temperature of thermal degradation was observed at 158°C and 124°C for the control and treated samples, respectively. Whereas, the end-set temperature of thermal degradation was found at 201°C and 178°C in the control and treated sample, respectively. The result showed about 21.52% decrease in the onset temperature and 11.44% decrease in the end-set temperature in treated sample with respect to the control. The percent weight loss during thermal decomposition was 63.41% in the control and 59.18% in the treated sample. It showed the decrease in % weight loss during thermal decomposition after biofield treatment. Based on this, it is presumed that biofield treated MNE may be more stable as compared to the control. The DTG thermogram exhibited the T max (the temperature at which the sample lost its maximum weight) at 177.81°C in the control sample and at 153.78°C in the treated sample of MNE. The result revealed about 13.51% decrease in T max of treated sample with respect to the control. This decrease in T max in treated sample might be due to increase in vaporization or volatilization [ 32] of treated MNE molecules after biofield energy treatment. It might be correlated with the alteration in internal energy through biofield energy treatment that results into earlier vaporization in treated sample as compared to the control.
FT-IR spectroscopic analysis
FT-IR spectra of the control and treated samples of MNE ( Figure 6) were inferred using the theoretically predicted wavenumber. The aromatic =C-H stretching peak was assigned at 3007-3057 cm⁻¹ in all three samples i.e., the control and treated (T1 and T2). Likewise, the -C-H (methyl) stretching was attributed to peak at 2962 cm⁻¹ in all three samples i.e., the control and treated (T1 and T2). The aromatic C=C stretching of naphthyl moiety was appeared in the region of 1508- 1631 cm⁻¹ in all the control and treated samples. The C-H asymmetrical bending peaks were observed at 1462-1475 cm⁻¹ in the control and T1 sample; whereas, this peak was observed at 1452-1475 cm⁻¹ region in T2 sample. In addition, the C-H symmetrical bending peak was observed at 1350 cm⁻¹ in all three samples. The C-O stretching for ether linkage was observed at 1030 cm⁻¹ in the control and the T2 sample while 1031 cm⁻¹ in the T1 sample. The =C-H in-plane deformation peaks were observed at the frequency region of 1172-1259 cm⁻¹, whereas the C-H out of plane deformation peaks were observed at 742-837 cm⁻¹ in all three samples. The observed FT-IR spectra were well supported with the literature data [ 33].
Uv-Vis spectroscopy
The UV spectra of control and treated (T1 and T2) samples are shown in Figure 7. The UV spectrum of control MNE exhibited the three different absorption maxima (λ max) at 225.4, 270.4, and 327.2 nm. The Uv spectrum of T1 sample showed the similar pattern of λ max i.e., at 224.4, 270.4, and 327.2 nm. Whereas, the T2 sample sample exhibited the λ max at 233.0, 270.5, and 327.0 nm. The result indicated the shifting of λ max in T2 sample (233.0 nm) as compared to control (225.4 nm). Although, no change in λ max was observed in T1 sample of MNE. Overall, the UV-vis spectral analysis suggests that biofield energy treatment may not cause any significant alterations in the λ max of treated MNE samples with respect to the control.

Conclusion

In brief, the XRD diffractogram of biofield treated MNE exhibited the decrease in intensity of XRD peaks as well as the decrease in average crystallite size (30.70%) as compared to the control. The surface area analysis showed an increase (5.32%) in surface area of biofield treated MNE with respect to the control. The thermal analysis (DSC, TGA/ DTG) showed a decrease in T max by 13.51%. Moreover, the latent heat of fusion was slightly increased by 0.83% in treated sample as compared to the control.

Overall, it can be concluded that Mr. Trivedi's biofield energy treatment has the impact onphysicochemical and thermal properties of treated MNE with respect to the control. Based on this, it is assumed that biofield treated MNE could be more useful chemical intermediate in the organic synthesis of various pharmaceutical drugs.

Acknowledgement

The authors like to acknowledge the Trivedi Science, Trivedi Master Wellness and Trivedi Testimonials for their steady support during the work. Authors would also like to thanks the whole team from the MGV pharmacy college, Nashik for providing the instrumental facility.

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20. Trivedi MK, Patil S, Shettigar H, Bairwa K, Jana S (2015) Spectroscopic characterization of biofield treated metronidazole and tinidazole. Med Chem 5: 340-344.

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# data[].attributes.attachment_link.keywords[]
methyl-2-naphthyl ether; biofield energy; x-ray
diffraction; surface area analysis; differential scanning calorimetry;
thermogravimetric analysis

# data[].attributes.attachment_link.title
Physical, Thermal, and Spectroscopic Characterization of Biofield Energy Treated Methyl-2-Naphthyl Ether

# data[].attributes.attachment_link.description
Mahendra Kumar Trivedi1 , Alice Branton1 , Dahryn Trivedi1 , Gopal Nayak1 , Khemraj Bairwa2 and Snehasis Jana2 * 1 Trivedi Global Inc., 10624 S Eastern Avenue Suite A-969, Henderson, NV 89052, USA 2 Trivedi Science Research Laboratory Pvt.

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		Impact of Biofield Treatment on p-Nitroaniline
	

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		Impact of Biofield Treatment on p-Nitroaniline
	

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| Para nitroaniline is an organic compound, used as an intermediate in the synthesis of pharmaceutical drugs, gasoline and dyes. Read this report to evaluate the impact of biofield treatment on the spectroscopic and physicochemical properties of p-Nitroaniline.

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Impact of Biofield Treatment on Spectroscopic and Physicochemical Properties of p-Nitroaniline | Insight Medical Publishing

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Impact of Biofield Treatment on Spectroscopic and Physicochemical Properties of p-Nitroaniline, Mahendra Kumar Trivedi, Alice Branton, Dahryn Trivedi, G

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		Biofield | Impact on Citrobacter braakii: A Urinary Pathogen
	

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		Biofield | Impact on Citrobacter braakii: A Urinary Pathogen
	

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| Citrobacter braakii is widespread in nature, mainly found in human urinary tract. The present study was designed to investigate the impact of biofield treatment on C. braakii in lyophilized as well as revived state for antimicrobial susceptibility pattern, biochemical characteristics, and biotype number.

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		Staphylococcus aureus after An Impact of Biofield Treatment
	

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		Staphylococcus aureus after An Impact of Biofield Treatment
	

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| The study conducted by Mahendra Kumar Trivedi and research team to evaluate the antibiogram, biochemical reactions and genotyping characterization of human biofield energy treated staphylococcus aureus.

# data[].attributes.attachment_link.keywords[]
staphylococcus aureus

# data[].attributes.attachment_link.title
Antibiogram, Biochemical Reactions and Genotyping Characterization of Biofield Treated

# data[].attributes.attachment_link.description
Staphylococcus aureus (S. aureus) is the key organism for food poisoning due to massive production of heat stable exotoxins. The current study was attempted to investigate the effect of Mr. Trivedi's biofield treatment on S. aureus. S. aureus (ATCC 25923) was divided into two parts, Group (Gr.) I: control and Gr.

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		Evaluation of Biofield Energy Treated Nocardia otitidis
	

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		Evaluation of Biofield Energy Treated Nocardia otitidis
	

# data[].attributes.body
| Nocardiosis is a soil-borne aerobic infection caused by Nocardia species commonly affects the respiratory tract. A study was performed to evaluate the antibiogram, genotype and phylogenetic analysis of biofield energy treated Nocardia otitidis.

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Nocardia otitidis; Nocardiosis; Antimicrobial susceptibility; Biofield energy treatment; 16S rDNA sequencing; Random amplified 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 Amplified Polymorphic DNA; CNS: Central Nervous System

The 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 differ from the mycobacteria by the possession of short-chain (40 to 60 carbons) mycolic acids [ 1]. Nocardia otitidis ( N. otitidis) is a weak filamentous Gram-positive, catalase-positive, branching rods shaped bacterium that appears similar to Actinomyces species. However, it can usually be differentiated from Actinomyces by acid-fast staining [ 2]. The 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. The 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 find any difference between two genera [ 4]. Based on immunoblot and enzyme-linked immunosorbent assay (ELISA) techniques that detect specific antibodies that appear as common in various Nocardia and Actinomadura species. These 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]. The most common manifestation of Nocardial disease is pulmonary nocardiosis [ 7], extrapulmonary disease [ 8], and ocular 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 often used as the drug of choice for the treatment of nocardiosis [ 12]. Therefore, some alternative strategies are needed to treat against nocardiosis. Biofield 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 biofield (putative energy fields) 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 significance [ 14]. Recently, it was discovered that all the electrical processes happening in the human body have strong relationship with the magnetic field as required by Ampere's law, which states that the moving charge produces magnetic fields in the surrounding space [ 15, 16]. Thus, 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 biofield. Therefore, the biofield consists of an electromagnetic field, being generated by moving electrically charged particles (ions, cell, molecule, etc.) inside the human body [ 17]. The 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 scientific instruments such as Kirlian photography, polycontrast interference photography (PIP) and resonance field imaging (RFI) can be extensively used to measure the biofield of human body [ 19]. Thus, 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. The objects always receive the energy and responding into the useful way that is called biofield energy and the process is known as biofield energy treatment. Mr. Mahendra Trivedi's unique biofield energy treatment (The Trivedi Effect ®) 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 significance of N. otitidis and significant impact of Mr. Trivedi's biofield energy modality on microbes, the present work was undertaken to evaluate the impact of Mr. Trivedi's biofield energy on N. otitidis in relation to antimicrobials susceptibility, minimum inhibitory concentration (MIC), random amplified polymorphic DNA analysis (RAPD) and 16S rDNA sequencing.

N. otitidis, American Type Culture Collection (ATCC 14630) strain was procured from Bangalore Genei, Bangalore-India in two vials A and B. Two different sealed packs were stored with proper storage conditions until further use. All the tested antimicrobials were procured from Sigma-Aldrich, India. The 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]. The 16S rDNA sequencing and DNA fingerprinting (RAPD) studies were carried out using Ultrapure Genomic DNA Prep Kit; Cat KT 83 (Bangalore Genei, India).

Experimental design and biofield treatment strategy

N. otitidis strain was divided into two groups, i.e., control and treated. The treated group was in sealed pack and handed over to Mr. Trivedi for biofield energy treatment under laboratory conditions. Mr. Trivedi provided the treatment through his energy transmission process to the treated group without touching the sample. After 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. These two biofield 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. The result of treated sample was compared with respect to the control. The 16S rDNA analysis was performed on biofield treated samples A and its subcultured sample A1.

Antimicrobial susceptibility test

The antimicrobial susceptibility testing is important for clinically significant species. The 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 nontuberculous mycobacteria (CLSI, M24-A, 2003) [ 29]. The detailed experimental procedure and conditions were followed as per the manufacturer's instructions. The 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 amplified polymorphic DNA (RAPD) analysis

For DNA fingerprinting (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 five RAPD primers, which were labelled as RBA8A, RBA13A, RBA20A, RBA10A and RBA15A were adopted from earlier studies. The PCR mixture contained 2.5 μL each of buffer, 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 amplification 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 final extension at 72°C for 7 min. Amplified 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]. The 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.

Amplification and gene sequencing of 16S rDNA

Genomic DNA was isolated from N. otitidis cells (set A, sample coded as 9A) using genomic purification kit, according to the manufacturer instructions. 16S rDNA gene (~ 1.5 kb) fragment was amplified with the help of high-fidelity polymerase chain reaction (PCR) using universal primers; forward primer (5'-AGAGTTTGATCCTGGCTCAG-3') and reverse primer (3'-ACGGTCATACCTTGTTACGACTT-5'). Amplified 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). The PCR amplified fragment was purified from the agarose gel using a DNA gel extraction kit. Sequencing of amplified product was done on a commercial basis from Bangalore Genei, India. The 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 software [ 31].

Results and Discussion

Antimicrobial susceptibility test

Antimicrobial susceptibility pattern of Nocardia can vary from species to species. The therapeutic effectiveness depends on the proper identification of species in infected patients and on in vitro sensitivity studies [ 32]. The 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]. The outcome of N. otitidis susceptibility pattern and MIC values of tested antimicrobials after biofield energy treatment are summarized in Table 1 and 2 respectively. The 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. The treated cells of N. otitidis did not show any alteration with respect to antimicrobial susceptibility pattern as compared to the control (Table 1). The susceptible nature of N. otitidis to amikacin, sulfamethoxazole, and ciprofloxacin and resistance pattern to ceftriaxone, amoxicillinclavulanic 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 biofield 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 after the biofield 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 twofold in treated sample after Mr. Trivedi's biofield energy treatment. Hence, authors assumed that this improvement of MIC value without alteration of sensitivity pattern may be due to the effect of putative energy transmit through biofield healing (Table 2).

Random amplified polymorphic DNA (RAPD) analysis

The treated and control samples were identified on the basis of their different and discriminative RAPD patterns. RAPD is a preferred tool that is being used now days to correlate the genetic similarity or mutations between species. The 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 efficiently detected using the PCR primers and identify inter-strain variations among species in treated samples [ 34]. The degree of relatedness and genetic mapping can be correlated between similar or different treated sample species [ 35].

The DNA fingerprinting by RAPD analysis using five primers was carried out on the control and treated samples. DNA fingerprinting by RAPD analysis of the control and treated samples are shown in Figure 1, and the polymorphic bands are marked by arrows. The RAPD patterns of treated samples showed some unique and dissimilar patterns. DNA polymorphism analyzed by the RAPD analysis was presented in Table 3. The level of polymorphism between control and treated samples (A, A1, B, and B1) are summarized in Table 4. The level of polymorphism was found in an average range of 34 to 53% in treated samples as compared to control in N. otitidis after the biofield treatment. The 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).

The bacteria that are poorly differentiated by conventional methods needs molecular analysis method like 16S rDNA sequence [ 36]. This molecular-based technique is a suitable tool for identification of most 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-specific variability [ 37]. The 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. The 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. The nearest homolog genusspecies 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. The distance matrix based on nucleotide sequence homology data are presented in Table 6. The phylogenetic tree was established using BLAST-Webpage (NCBI). According to Table 6, ten different related bacterial species of N. otitidis were selected as Operational Taxonomic Units (OTUs) in order to investigate the phylogenetic relationship of N. otitidis. There were 1462 base-pair nucleotides of 16S rDNA gene sequences, which were analyzed and multiple alignments were constructed using ClustalW in MEGA3.1 [ 31]. The 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. This 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. The 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. The results suggested that after biofield treatment 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 findings, authors assumed the sustainability of The Trivedi Effect ® 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 effective 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. Biofield 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 different phenotypic characteristics [ 38]. Based on these results, it is expected that biofield treatment has the scope to be a cost effective and alternative approach than the existing antimicrobial therapy in near future (Table 5 and 6).

Altogether, the biofield 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 interspecific relationships with N. otitidis after biofield treatment. The 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). The results suggest that there is an impact of biofield treatment on MIC, 16S rDNA analysis, and DNA polymorphism of N. otitidis. These changes were found in the organism may be due to alterations happened at the genetic level after biofield treatment. Overall, it seems that Mr. Trivedi's unique biofield energy treatment might be used as an alternate treatment approach in future antimicrobial therapy.

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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21. Lenssen AW (2013) Biofield and fungicide seed treatment influences on soybean productivity, seed quality and weed community. Agricultural Journal 83: 138-143.

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# data[].attributes.attachment_link.title
Evaluation of Antibiogram, Genotype and Phylogenetic Analysis of Biofield Treated Nocardia otitidis

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Nocardia otitidis; Nocardiosis; Antimicrobial susceptibility; Biofield energy treatment; 16S rDNA sequencing; Random amplified 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 Amplified Polymorphic DNA; CNS: Central Nervous System 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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		Antibiogram Typing of Biofield Energy Treated Staphylococcus Species
	

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		Antibiogram Typing of Biofield Energy Treated Staphylococcus Species
	

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| The present work was designed to investigate the impact of biofield energy treatment on MDR strains of Staphylococcus species (S. haemolyticus, S. epidermidis and S. aureus) with respect to antibiogram typing, biochemical reaction pattern, followed by biotyping.

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Antibiogram Typing of Biofield Treated Multidrug Resistant Strains of

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Antimicrobial resistance is a global health issue in the developing countries. This study was carried out to evaluate the impact of Mr. Trivedi's biofield energy treatment on multidrug resistant (MDR) clinical lab isolates (LSs) of Staphylococcus species viz. Staphylococcus haemolyticus (LS 18), Staphylococcus epidermidis (LS 21), and Staphylococcus aureus (LS 30).

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		Assessment of Enterobacter aerogenes Isolates after Biofield Treatment
	

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		Assessment of Enterobacter aerogenes Isolates after Biofield Treatment
	

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| Enterobacter is a genus of gram-negative, rod shaped, facultative anaerobic, and non-spore forming microbes of family Enterobacteriaceae. The study to analyze the assessment of antibiogram of multidrug-resistant isolates of Enterobacter aerogenes after biofield energy treatment.

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Enterobacter aerogenes; Multidrug resistant; Antimicrobial susceptibility; Biofield treatment; Biochemical reactions; Biotyping

Enterobacter is a genus of Gram-negative, rod shaped, facultative anaerobic, and non-spore forming microbes of family Enterobacteriaceae. Enterobacter aerogenes ( E. aerogenes) is well known opportunistic bacteria emerged as nosocomial pathogen in intensive care unit patients [ 1]. E. aerogenes was initially named as Aerobacter aerogenes, which was later in 1960 included in the genus Enterobacter. Since 1990s, E. aerogenes has been increasingly reported for resistant against different antimicrobials, leads to emergence of multidrugresistant (MDR) isolates [ 2]. In the last 5 years, clinical isolates of this species have shown natural resistant against aminopenicillins, often showed resistance against β-lactams antibiotics. Resistance mechanisms in β-lactams mostly involve enzymatic degradation and plasmid-mediated broad spectrum β-lactamases [ 3]. However, membrane permeability, enzyme degradation, and p-glycoprotein efflux pump also contribute it to enhance the level of resistance against carbapenems, fluoroquinolones, quinolones, tetracycline, and chloramphenicol [ 4, 5]. Enterobacter species are responsible for high morbidity and mortality rate in recent years due to nosocomial infections and other health care settings [ 6]. Due to extended resistance of Gram-negative bacteria against almost all antibiotics, early initiation of drug therapy is required, nowadays colistin, and polymyxin antibiotic have been preferred as an alternative drugs against Gramnegative pathogens [ 7, 8]. Recent update on colistin antibiotic, a drug of 21 st century reports it's associated adverse effects and serious toxicity issues such as neurotoxicity and nephrotoxicity [ 9]. Despite several new drug discoveries of broad spectrum drugs or combination therapies, associated toxicities are still a serious complication. Recently, an alternate treatment therapy approach called biofield healing therapies or therapeutic touch is reported with effectively inhibiting the growth of bacterial cultures [ 10].

The biofield is a cumulative outcome of measurable electric and magnetic field, exerted by the human body [ 11]. It generates through some internal processes in the human body such as blood flow, lymph flow, brain functions, and heart function. The energy mainly exists in different forms such as potential, kinetic, magnetic, electrical, and nuclear energy produced from different sources. The energy field that surrounds and penetrates the human body is collectively defined as biofield and the extent of energy associated with biofield is termed as biofield energy. Biofield treatment includes energy therapies that interact with patient's biofield and lead to improve people's health and wellbeing. Mr. Trivedi's possesses unique biofield energy, which has been experimentally studied in various research fields. Mr. Trivedi's unique biofield treatment is also known as The Trivedi Effect ®. Recently, Mr. Trivedi's biofield has made significant breakthrough and results in living organisms and nonliving materials in a different manner. In life sciences, biofield treatment has altered the antimicrobial sensitivity pattern of pathogenic microbes. In some bacteria, genus as well as species was found to be altered [ 12- 14]. Mr. Trivedi's biofield treatment has also well scientifically studied in different areas such as materials science research [ 15- 18], biotechnology research [ 19, 20], and agriculture research [ 21- 23]. Due to paucity of information and considering biofield energy as an alternate treatment approach, the present work was undertaken to evaluate the impact of Mr. Trivedi's biofield energy treatment on antimicrobials susceptibility, biochemical reactions pattern, and biotype of MDR isolates of E. aerogenes.

The Two clinical MDR lab isolates (LSs) of E. aerogenes (i.e., LS 45 and LS 54) were procured from stored stock cultures in microbiology lab, Hinduja hospital, Mumbai and stored as per suggested storage conditions until further use. The acceptability of the identification media and antimicrobial agents were checked prior to the study. The antimicrobials and biochemicals used in the study were procured from Sigma Aldrich, MA, USA. The antimicrobial susceptibility, biochemical reactions, and biotype number were evaluated on MicroScan Walk- Away ® (Dade Behring Inc., West Sacramento, CA) using Negative Breakpoint Combo 30 (NBPC 30) panel. The panels were allowed to equilibrate to room temperature prior to rehydration. All opened panels were used in same day.

The turbidity standard technique using direct inoculation of E. aerogenes was used. Using a sterile wooden applicator stick or bacteriological loop, the surface of 4-5 large or 5-10 small morphologically similar culture was touched for well-isolated colonies from an 18-24 hour non-inhibitory agar plate. Further, colonies were emulsified in 3 mL of inoculum water (autoclaved deionized water) to an equivalent of a 0.5 McFarland barium sulfate turbidity standard. 100 μL of the standardized suspension was pipetted into 25 mL of inoculum water using pluronic and inverted 8-10 times.

Treated group of E. aerogenes was subjected to biofield treatment, keeping the control group untreated. The treatment group in sealed pack was handed over to Mr. Trivedi for biofield treatment under laboratory condition. Mr. Trivedi provided the treatment through his energy transmission process to the treated groups without touching the samples. Treated samples were assessed for antimicrobial sensitivity, biochemical reactions, and biotyping as per experimental design. Whilst handing over these cultures to Mr. Trivedi for treatment purposes, optimum precautions were taken to avoid contamination.

Evaluation of antimicrobial susceptibility assay

Antimicrobial susceptibility pattern of MDR E. aerogenes clinical isolates were studied using MicroScan Walk-Away ® using NBPC 30 panel as per manufacturer's instructions. The antimicrobial susceptibility pattern (S: Susceptible, I: Intermediate, R: Resistant, and IB: Inducible β-lactamase) and minimum inhibitory concentration (MIC) values were determined by observing the lowest antimicrobial concentration showing growth inhibition [ 24]. Antimicrobials used in susceptibility and MIC assay viz. amikacin, amoxicillin/K-clavulanate, ampicillin/ sulbactam, ampicillin, aztreonam, cefazolin, cefepime, cefotaxime, cefotetan, cefoxitin, ceftazidime, ceftriaxone, cefuroxime, cephalothin, chloramphenicol, ciprofloxacin, gatifloxacin, gentamicin, imipenem, levofloxacin, meropenem, moxifloxacin, norfloxacin, nitrofurantoin piperacillin, piperacillin/tazobactam, tetracycline, ticarcillin/Kclavulanate, tobramycin, and trimethoprim/sulfamethoxazole.

Biochemical reaction study

Biochemical reactions of MDR isolates of E. aerogenes were determined by using MicroScan Walk-Away ® system in both control and treated groups. Biochemicals used in the study viz. acetamide, adonitol, arabinose, arginine, cetrimide, cephalothin, citrate, colistin, esculin hydrolysis, nitrofurantoin, glucose, hydrogen sulfide, indole, inositol, kanamycin, lysine, malonate, melibiose, nitrate, oxidationfermentation, galactosidase, ornithine, oxidase, penicillin, raffinose, rhamnose, sorbitol, sucrose, tartrate, tryptophan deaminase, tobramycin, urea, and Voges-Proskauer [ 24].

Identification by biotype number

The biotype number of MDR isolates of E. aerogenes control and treated sample were determined by MicroScan Walk-Away ® processed panel data report with the help of biochemical reaction data [ 24].

Antimicrobial susceptibility study

The antimicrobial susceptibility of control and treated MDR isolates of E. aerogenes are presented in Table 1. Biofield treatment in LS 45, showed altered sensitivity in three antimicrobials, i.e., cefotetan changed from I → IB, piperacillin/tazobactam sensitivity changed from R → IB, and tetracycline sensitivity changed from I → S. After biofield treatment, sensitivity of chloramphenicol and tetracycline were improved from R → I in LS 54. Biofield treatment on LS 45 showed 10.71%, while LS 54 showed 7.14% alterations among tested antimicrobials as compared to control (Figure 1). Overall, 14.28% antibiotics out of twenty-eight tested antimicrobials showed alteration in antimicrobial sensitivity assay with respect to control. Rest of the antimicrobials did not show any change in sensitivity pattern in clinical isolates as compared to their respective control.

Determination of minimum inhibitory concentration (MIC)

MIC values of all antimicrobials in control and biofield treated clinical MDR isolates of E. aerogenes are summarized in Table 2. Biofield treatment in LS 45, significantly reduced the MIC values of cefotetan, piperacillin/tazobactam, and tetracycline antimicrobials as compared to control. Chloramphenicol (16 μg/mL) and tetracycline (8 μg/mL) also showed decreased MIC values in LS 54 with respect to control. Four-fold decrease was observed in piperacillin/tazobactam (≤ 16 μg/mL) and two-fold decreases were found in cefotetan (≤ 16 μg/ mL) and tetracycline (≤ 4 μg/mL). Biofield treatment showed alteration in three antimicrobials MIC value in LS 45 (9.37%), while MIC of two antimicrobials showed alteration in LS 54 (6.25%) with respect to control (Figure 1). An overall 12.50% among tested antimicrobials out of thirty two showed alteration in MIC values. Rest of the antimicrobials did not show any alteration in MIC values after biofield treatment in both clinical isolates of E. aerogenes.

Biochemical and biotype number study

Biochemical study results of control and biofield treated isolates of E. aerogenes are summarized in Table 3. Results showed an overall 45.45% reverse reactions in tested biochemical as compared to control. LS 45 showed only 3.03% alteration, i.e., negative reaction (positive '+' to negative '-') in arginine, while in LS 54, showed 42.42% negative reaction after biofield treatment as compared to control (Figure 1).

Adonitol, arabinose, esculin hydrolysis, glucose, inositol, melibiose, oxidation-fermentation, galactosidase, raffinose, rhamnose, sorbitol, sucrose, tartrate, and Voges-Proskauer showed negative reaction after biofield treatment in LS 54. Rest of the biochemicals did not show any change in reaction with respect to the control.

Based on the above results of biochemical reactions, significant alteration in biotype number was observed in both isolates after biofield treatment as compared to control. In LS 45, changed biotype was found as 7774 5372, as compared to control biotype number 7774 7372. New organism was identified as Stenotrophomonas maltophilia with brown color as special characteristics along with altered biotype number 0406 0374 after biofield treatment in LS 54 on day 10 with respect to control, 7770 4376 (Table 4).

Biofield treatment was reported as an alternative therapy in medical health care practice [ 25]. This experimental was designed to demonstrate the influence of biofield treatment on MDR isolates of E. aerogenes for its susceptibility pattern, biochemical reaction and biotype number. The emergence of MDR isolates of E. aerogenes harbored a global health problem and an emerging Gram-negative MDROs commonly associated with severe systemic and hospital acquired infections in human. MDR is an unavoidable natural phenomenon which results due to continuous discovery of newer drugs. This experiment showed that, biofield treatment induces changes in susceptibility pattern of antimicrobials such as chloramphenicol, tetracycline, cefotetan, piperacillin/tazobactam, and tetracycline as compared to control. Tetracycline and chloramphenicol showed increased sensitivity after biofield treatment in LS 54 as compared to control. Clinical isolates of E. aerogenes have a broad ability to develop antimicrobial resistance [ 26]. Natural resistance against chloramphenicol may be mediated either enzymatically through acetylation of the drug or mechanically via active drug efflux [ 27]. However, efflux mechanism is generally involved to expel the antimicrobials such as tetracycline, fluoroquinolones and chloramphenicol in Enterobacter sp. [ 28]. Biofield treatment on clinical isolates of E. aerogenes might act on enzymatic level which may change the mechanism of resistant against chloramphenicol and tetracycline.

A significant decreased MIC values in cefotetan, piperacillin/ tazobactam, and tetracycline antimicrobials were found after biofield treatment in LS 45. Chloramphenicol and tetracycline also showed decreased in MIC values in LS 54 along with increases antimicrobial sensitivity with respect to control. Increased incidence of nosocomial infections and broad resistance against third generation cephalosporins, penicillins and quinolones is a serious problem. A number of newer agents so called "fourth generation" antimicrobials remain effective for treatment [ 29, 30]. Aminoglycosides, quinolones, trimethoprim/ sulphamethoxazole, and carbapenems displays good activity against Gram-negative pathogens including Enterobacter species [ 31, 32]. Biofield treatment significantly decreased the MIC values of piperacillin/tazobactam, cefotetan, tetracycline, and chloramphenicol, which could suggest the action of biofield treatment on enzymatic/ genetic level which could affect the β-lactamases production that may lead to decrease the minimum concentration of antimicrobials required to inhibit the in vitro growth of E. aerogenes.

Several phenotypic identification tests were available to differentiate the Enterobacter species. Experimental identification of E. aerogenes was performed using a series of biochemical analysis. Basic characteristics of Enterobacter species in biochemical reactions are presence of Voges- Proskauer, sucrose, dextrose, glucose, lactose, rhaminose, citrate, lysine, ornithine decarboxylase, and motile in nature. Indole, methyl red and hydrogen sulphide are the negative characteristics test of Enterobacter species. Enterobacter aerogenes is a common contaminant of vegetable matter which generally forms shiny colonies with entire margins and convex elevation [ 33]. Biochemical reactions of control MDR isolates of E. aerogenes were well supported with literature data [ 34]. Biofield treatment showed alteration, i.e., negative reaction in arginine biochemical in LS 45 and adonitol, arabinose, esculin hydrolysis, glucose, inositol, melibiose, oxidation-fermentation, galactosidase, raffinose, rhamnose, sorbitol, sucrose, tartrate and Voges-Proskauer in LS 54 as compared to control, which is the basic character of Enterobacter species. Various biochemical mechanisms are involved in biochemical, such as enzymatic alteration involved in sugar transferase, isomerization of biochemical, hydrolyzing reactions, etc. [ 35]. Biofield treatment on MDR isolates of E. aerogenes may alter the enzymatic biochemical reactions which could show result in altered biochemical reactions as compared to control.

Biotyping was also performed using an automated system and found a significant changed in biotype number in both isolates on day 10, and a new organism was identified as Stenotrophomonas maltophilia after biofield treatment in LS 54 as compared to control. The results of biotype number and identification of new species are based on the biochemical reaction pattern of control and treated samples. The biochemical reaction patterns of LS 54 after biofield treatment are well supported with literature data [ 36], which suggest the alteration in species as Stenotrophomonas maltophilia (Table 4). Based on this, biochemical reaction pattern, biotype number and identified species results are well collaborated.

In biomedical health care system, biofield therapies are very popularly used to enhance human wellbeing and helps in minimizing patient's health [ 25]. Biofield treatment might act as a communication system using electromagnetic frequencies which will carry message from environment to organism and vice versa [ 37]. However, National Center for Complementary and Alternative Medicine/National Institute of Health (NCCAM/NIH), now defined biofield therapies in subcategory of energy therapies as one of the five complementary medicine domains [ 25]. In microbiology, Mr. Trivedi's biofield treatment was reported on pathogenic microorganism which altered the phenotypic characteristics of microorganism [ 12- 14]. Alteration in antimicrobials sensitivity pattern after treatment might involve cell receptor protein and its interactions at molecular level. Experimental data showed that, biofield treatment include significant changes in susceptibility pattern of antimicrobials, biochemical reactions, MIC values, and biotype number. Biofield treatment could be an alternative approach to study the alteration in sensitivity pattern of MDR isolates.

Overall data illustrate that there has a significant impact of biofield treatment on antimicrobial susceptibility pattern, MIC values, biochemical reactions, and biotype number in clinical MDR isolates of E. aerogenes. Antimicrobial sensitivity assay showed 14.28% alteration, MIC values were significantly decreased, i.e., 12.50% among tested antimicrobials. Biochemical study showed an overall 45.45% altered reactions in tested biochemical as compared to control after biofield treatment in clinical isolates. On the basis of changed biotype number (0406 0374), new organism was identified as Stenotrophomonas maltophilia with brown color as special characteristics after biofield treatment in LS 54 as compared to control (7770 4376). Mr. Trivedi's biofield energy treatment could be applied to improve the sensitivity of antimicrobials, which may be an alternative therapeutic healing approach in medical science to fight against infections due to the emergence of multi drug-resistant strain of E. aerogenes.

Authors gratefully acknowledged the whole team of PD Hinduja National Hospital and MRC, Mumbai, Microbiology Lab for their support. Authors also would like to thanks Trivedi Science, Trivedi master wellness and Trivedi testimonials for their support during the work.

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15. Trivedi MK, Patil S, Tallapragada RM (2013) Effect of biofield treatment on the physical and thermal characteristics of vanadium pentoxide powder. J Material Sci Eng S11: 001.

16. Trivedi MK, Patil S, Tallapragada RM (2015) Effect of biofield treatment on the physical and thermal characteristics of aluminium powders. Ind Eng Manage 4: 151.

17. Nayak G, Altekar N (2015) Effect of biofield treatment on plant growth and adaptation. J Environ Health Sci 1: 1-9.

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20. Lenssen AW (2013) Biofield and fungicide seed treatment influences on soybean productivity, seed quality and weed community. Agricultural Journal 8: 138-143.

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# data[].attributes.attachment_link.title
Assessment of Antibiogram of Multidrug-Resistant Isolates of Enterobacter aerogenes after Biofield Energy Treatment

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Enterobacter aerogenes; Multidrug resistant ; Antimicrobial susceptibility; Biofield treatment; Biochemical reactions; Biotyping Enterobacter is a genus of Gram-negative, rod shaped, facultative anaerobic, and non-spore forming microbes of family Enterobacteriaceae. Enterobacter aerogenes ( E. aerogenes) is well known opportunistic bacteria emerged as nosocomial pathogen in intensive care unit patients [ 1].

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		Analyze The Impact of Biofield Treatment on Klebsiella pneumoniae
	

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		Analyze The Impact of Biofield Treatment on Klebsiella pneumoniae
	

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| The study conducted by Mahendra Kumar Trivedi and research team to investigate the antimicrobial susceptibility, biochemical characterization and molecular typing of human biofield energy treated Klebsiella pneumoniae. Follow the link to know more!

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Mahendra Kumar Trivedi1, Alice Branton1, Dahryn Trivedi1, Mayank Gangwar2 and Snehasis Jana2*

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, Madhya Pradesh, India

*Corresponding Author:

Dr. Snehasis Jana
Trivedi Science Research Laboratory Pvt. Ltd.
Hall-A, Chinar Mega Mall, Chinar Fortune City
Hoshangabad Rd., Bhopal-462026, Madhya Pradesh, India
Tel: +91-755-6660006
E-mail: [email protected]

Received date: September 15, 2015; Accepted date: September 22, 2015; Published date: September 25, 2015

Citation: Trivedi MK, Branton A, Trivedi D, Gangwar M, Jana S (2015) Antimicrobial Susceptibility, Biochemical Characterization and Molecular Typing of Biofield Treated Klebsiella pneumoniae. J Health Med Inform 6:206. doi:10.4172/2157-7420.1000206

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.

Visit for more related articles at Journal of Health & Medical Informatics

Keywords

Klebsiella pneumoniae; Biofield energy treatment; Antibiogram, Biochemical reactions, Polymorphism; Random Amplified Polymorphic DNA.

Abbreviations

CAM: Complementary and Alternate Medicine; NHIS: National Health Interview Survey; NCHS: National Center for Health Statistics; ATCC: American Type Culture Collection; MIC: Minimum Inhibitory Concentration; MEGA: Molecular Evolutionary Genetics Analysis; NBPC 30: Negative Breakpoint Combo Panel 30; RAPD: Random Amplified Polymorphic DNA; PCR: Polymerase chain reaction; ESBL: Extended Spectrum β-Lactamase

Introduction

The increased medical practice for antibiotic usage creates selection pressure and results emergence of nosocomial pathogens. Klebsiella pneumoniae ( K. pneumoniae ) is a Gram-negative, facultative anaerobic and rod-shaped bacterium of the Enterobacteriaceae family. It is regarded as an opportunistic pathogen that is associated with the hospital-acquired urinary tract infections, septicemia, pneumonia, and soft tissue infections [ 1]. K. pneumoniae is responsible for the nosocomial outbreaks worldwide, due to its ability to spread rapidly in the hospital environment [ 2], and results in high morbidity and mortality [ 3]. It has acquired resistance against extended-spectrum cephalosporins and penicillins, due to the production of extendedspectrum β-lactamases (ESBLs) [ 4].

Multidrug combination therapy and some alternate treatment options are required to control the infections associated with this microorganism. Due to the associated side effects and failure of drug treatment therapy, alternate and complementary therapy approach are the preferred treatment strategies. Recently, an alternate treatment approach using healing therapy or therapeutic touch known as biofield energy treatment, which has been widely reported in various research field. The biofield therapies (putative energy fields) were reported to alter the sensitivity of antimicrobial against treated microorganism [ 5], inhibits the growth of bacterial cultures [ 6], effect on in vitro cells, tissues [ 7], animals [ 8], and the clinical effects such as hematologic [ 9], immunologic effects [ 10], healing rates of wounds [ 11], etc. Biofield is the name given to the electromagnetic field that permeates and surrounds living organisms [ 12]. It is referred as the biologically produced electromagnetic and subtle energy field that provides regulatory and communication functions within the human organism. Specific environmental frequencies, are absorbed by the different biomolecules, due to changes in the movements of component parts. Therefore, the human or any living object, not only radiate but also absorb and respond to these frequencies [ 13]. Mr. Mahendra Kumar Trivedi is well known biofield treatment practitioners, and his unique biofield energy treatment is known as The Trivedi Effect ®. Mr. Trivedi's biofield energy treatment has been well known and studied in the field of materials science research [ 14- 16], agricultural science research [ 17, 18], and microbiology research [ 19, 20].

Due to the clinical importance of K. pneumoniae and outstanding results of biofield treatment, the present study was designed to evaluate the impact of Mr. Trivedi's biofield energy treatment on K. pneumoniae with respect to the antimicrobial susceptibility, biochemical study, and biotype number. Further, in order to study the phenotypic characteristics of biofield treated K. pneumoniae , molecular typing using arbitrary amplification of polymorphic DNA sequences, termed as random amplified polymorphic DNA (RAPD) analysis was used [ 21]. RAPD is a preferred technique used in different studies for typing and discriminating the epidemiology of microorganism [ 22]. RAPD has an advantage over other traditional phenotypic typing methods as it is rapid, relatively inexpensive and technically feasible [ 23]. The aim of this study was to evaluate the impact of Mr. Trivedi's biofield energy treatment on K. pneumoniae with respect to antibiogram characteristics and genotyping using RAPD of the organism.

Materials and Methods

K. pneumoniae ATCC 15380 [American Type Culture Collection] was procured from Bangalore Genei, in sealed pack, and stored as per the recommended storage conditions for further use. The antimicrobial susceptibility, minimum inhibitory concentration (MIC), biochemical reactions, and biotype number were evaluated using automated MicroScan Walk-Away ® system (Dade Behring Inc., West Sacramento, CA) using Negative Breakpoint Combo 30 (NBPC 30) panel. RAPD was carried out using Ultrapure Genomic DNA Prep Kit; Cat KT 83 (Bangalore Genei, India). All the tested antimicrobials, biochemicals, media, and reagents were procured from Sigma-Aldrich, India.
Biofield treatment modalities
K. pneumoniae strain was divided into two groups i.e. control and treated. The treated group was in sealed pack and handed over to Mr. Trivedi for the biofield energy treatment under laboratory conditions. Mr. Trivedi provided the treatment through his energy transmission process to the treated group that includes bioenergy emission of certain wavelength, which has the ability to do the changes in the microbes without touching the sample. Mr. Trivedi's unique energy treatment is known as The Trivedi Effect ®. Mr. Trivedi visited the laboratory individually over a period of treatment and for control experiments, nobody entered the experimental room during the treatment period. Whilst handing over these cultures to Mr. Trivedi for treatment purposes, optimum precautions were taken to avoid contamination. After treatment, control and treated groups were assessed on day 10 for the antimicrobial susceptibility, minimum inhibitory concentration (MIC), biochemical reactions, biotype, and genotyping using RAPD analysis. The result of treated sample was compared with respect to the control.
Investigation of antimicrobial susceptibility assay
Investigation of antimicrobial susceptibility of K. pneumoniae was carried out with the help of automated instrument, MicroScan Walk- Away ® using Negative Breakpoint Combo 30 (NBPC30) panel as per the manufacturer's instructions. The panel was allowed to equilibrate to room temperature prior to rehydration. All opened panel were used on the same day. The tests were carried out on MicroScan, which were miniaturized of the broth dilution susceptibility test that had been dehydrated. Briefly, 100 μL of the standardized suspension of K. pneumoniae was pipetted into 25 mL of inoculum water using pluronic and inverted 8-10 times and inoculated, rehydrated, and then subjected to incubation for 16 hours at 35°C. Rehydration and inoculation were performed using the RENOK ® system with inoculators-D (B1013-4). The detailed experimental procedures and conditions were followed as per the manufacturer's instructions. Briefly, after inoculation and rehydration with a standardized suspension of K. pneumoniae , it was incubated at 35°C for 16 hours. MIC and a qualitative susceptibility like susceptible (S), intermediate (I), and resistant (R) were determined by observing the lowest antimicrobial concentration showing growth inhibition [ 24].
Biochemical studies
The biochemical reactions of K. pneumoniae were determined by MicroScan Walk-Away ® where, interpretation of biochemical reactions for microbial identification of Gram-negative organisms [ 24].
Biotype number
The biotype number of K. pneumoniae was determined by MicroScan Walk-Away ® processed panel data utilizing data of biochemical reactions [ 24].
Random Amplified Polymorphic DNA (RAPD) analysis
Three series of inoculums (one for control and other two for treatment named as treated A and B) were prepared from K. pneumoniae sample. Two inoculums (treated samples A and B) were subjected to Mr. Trivedi's biofield energy treatment. Whilst handing over treated groups to Mr. Trivedi for biofield treatment, optimum precautions were taken to avoid the contamination. After that, the 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. Control and treated samples were incubated at 37°C with 160 rpm for 18 h. Subsequently, the cultures were spun down, and genomic DNA was isolated for control and treated samples using the genomic DNA Prep Kit (Bangalore Genei, India). The RAPD was performed with all samples of K. pneumoniae using five RAPD primers, which were labelled as RBA 5A, RBA 10A, RBA 15A, RBA 21A, and RBA 22A. The PCR mixture contained 2.5 μL each of buffer, 4.0 mM each of dNTP, 2.5 μM each of primer, 5.0 μL each of genomic DNA, 2U each of Taq polymerase, 1.5 μL of MgCl2 and 9.5 μL of nuclease-free water in a total of 25 μL mixture. PCR amplification protocol was followed with initial denaturation at 94ºC for 7 min, followed by 8 cycles of denaturation at 94ºC for 1 min, annealing at 35ºC for 1 min, and extension at 72ºC for 2 min; and 35 cycle of denaturation at 94ºC for 1 min, annealing at 38ºC for 1 min, and extension at 72ºC for 1.5 min; and the final extension at 72ºC for 7 min. Amplified PCR products (12 μL) from all the samples (control and treated) were separated on 1.5% agarose gels at 75 volts, stained with ethidium bromide and visualized under UV illumination [ 25].

The 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.

Results and Discussion
Antimicrobial susceptibility assay
The results of biofield treatment on K. pneumoniae with respect to antimicrobials susceptibility pattern and MIC are summarized in Table 1 and 2, respectively.

Table 1: Effect of biofield treatment on K. pneumoniae for its antimicrobial susceptibility

MIC values are presented in µg/mL;ESBL: Suspected extended-spectrum β-lactamases a, b screen

Table 2: Minimum inhibitory concentration (MIC) of tested antimicrobials against

The antimicrobial sensitivity result of three antimicrobials namely ampicillin/sulbactam, cephalothin, and chloramphenicol showed the alteration after biofield treatment with respect to control among twenty-eight tested antimicrobials. The sensitivity of ampicillin/ sulbactam and chloramphenicol was improved i.e. from intermediate (I) to susceptible (S), while cephalothin showed an altered sensitivity nature from S to I. Other tested antimicrobials did not show any alterations of sensitivity pattern as compared to the control.

MIC results were well supported with antimicrobial sensitivity data, as ampicillin/sulbactam and chloramphenicol showed decrease value of MIC after the biofield treatment. Ampicillin/sulbactam ( i.e. 16/8 to ≤8/4 μg/mL) and chloramphenicol ( i.e. 16 to ≤8 μg/mL) showed two folds change in MIC values as compared to the control. Cephalothin showed an alteration in MIC value i.e. from ≤8 to 16 μg/mL after biofield treatment. The rest of the tested antimicrobials did not show any alteration in MIC values with respect to the control.

Efficacy of sulbactam, a β-lactamase inhibitor, in combination with ampicillin, was well reported and a preferred treatment option against β-lactam-resistant K. pneumoniae infections. According to Hoffman et al. the combination of ampicillin/sulbactam was found to have synergistic effects, which significantly decreased the severity of pneumonia. Bronchoalveolar lavage cytologic findings, and extent of macroscopic lesions in lung tissue of the noninoculated regions were reported as compared to the individual ampicillin or sulbactam [ 26]. The resistance pattern of ampicillin against K. pneumoniae is because of two types of chromosomal β-lactamase enzymes being SHV-1 and LEN-1 [ 27], and results suggest that biofield treatment might alter these enzymes and alter the sensitivity pattern. Biofield treatment on K. pneumoniae has improved the sensitivity profile of ampicillin/ sulbactam and showed decreased MIC value by two-folds, which might be useful in the future treatment strategy against pneumoniae lung infection. Multi-drug therapies are another approach against β- lactamase producing strain of K. pneumoniae , as synergy has been frequently reported in vitro between β-lactams and aminoglycosides. According to Jones, a synergistic effect was frequently reported in a combination of ertapenem and ciprofloxacin [ 28]. Besides synergistic effect, antibiotics such as amikacin, gatifloxacin, gentamicin, and chloramphenicol are the preferred drug of choice with respect to cost, side effects and many other factors in K. pneumoniae associated infections [ 29]. Mr. Trivedi's biofield energy treatment on K. pneumoniae showed improved sensitivity pattern of chloramphenicol and simultaneously decreased the MIC value as compared to the control. β-lactamases are enzymes that inactivates the β-lactam containing antibiotic which is present in almost all Gram-negative bacilli such as E. coli and Klebsiella spp. [ 30]. Biofield treatment might alter the production of these enzymes which may result in the improved sensitivity of antimicrobials.
Organism identification by biochemical reactions
Biochemical study results of control and biofield treated groups are summarized in Table 3.

Table 3: Effect of biofield treatment on K. pneumoniae to the vital processes occurring in living organisms -: negative; +: positive.

The results showed overall 9.09% of tested biochemical altered reactions as compared to the control. The colistin and nitrate biochemical showed negative reaction i.e. from (+) positive to (-) negative as compared to the control. Cephalothin showed positive reaction i.e. from (-) negative to (+) positive reaction. The rest of tested biochemicals did not show any alteration in their reaction pattern after biofield treatment. The biochemical reactions of control K. pneumoniae were well supported with literature data [ 31].
Organism identification by biotype number
Based on the biochemical results, alteration in biotype number was also observed in the biofield treated K. pneumoniae as compared to the control. The control group showed biotype number as 7774 4274, while after treatment altered biotype number was reported as 7774 4272. However, no change in organism was reported after biofield treatment with respect to the control. Our research group recently reported the significant alterations in biochemical reactions followed by the change in biotype number that was also supported with published data [ 19, 20]. Biofield treatment might alter some enzymatic reactions in K. pneumoniae , which resulted in alterations in characteristic biochemical reactions and biotype number.
Random Amplified Polymorphic DNA (RAPD) analysis
RAPD has been used as a genotyping tool to that is being used to correlate the genetic similarity or mutations between species of K. pneumoniae [ 32]. DNA fingerprinting analysis of the control and treated K. pneumoniae was identified on the basis of their different and discriminative RAPD patterns. The simplicity and wide applicability of RAPD analysis mainly depend on the use of short nucleotide primers, which were unrelated to known DNA sequences of the target organism [ 21]. DNA polymorphism can be efficiently detected using the PCR primers and identify the inter-strain variations among species in the treated samples. The degree of relatedness and genetic mapping can be correlated between similar or different treated sample species [ 33].

In this experiment, DNA fingerprinting of control and treated samples using RAPD are shown in Figure 1.

The polymorphic bands are marked by arrows in the gel image. The RAPD patterns of treated samples showed some unique, dissimilar, common, and polymorphic patterns. DNA polymorphism among the different treated samples compared with the control were analyzed and presented in Table 4.

Table 4: DNA polymorphism of K. pneumoniae analyzed by random amplified polymorphic DNA (RAPD) analysis, TSA: treated sample A; TSA-1: treated sample A-1; TSB: treated sample B; TSB-1: treated sample B-1.

The level of polymorphism between control and treated samples (A, A1, B, and B1) are summarized in Table 5.

Table 5: Level of polymorphism between control and treated K. pneumoniae samples. C: Control; TSA: treated sample A; TSA-1: treated sample A-1; TSB: treated sample B; TSB-1: treated sample B-1.

The level of polymorphism was found in an average range of 11 to 15% in the treated samples as compared to control after the biofield treatment. The highest change in DNA sequence was observed in treated samples with RBA 10A primer as compared to control; whereas no change was found in treated sample with RBA 21A primer as compared to control. Thus, results indicates that treatment samples has genetic variability among organism. RAPD also explains the relevant degree of genetic diversity, however this technique has the potential to detected genetic polymorphism throughout the genome [ 34].

Biofield energy as the complementary medicine is well documented and considered as alternate medicine approach worldwide. According to the report of National Health Interview Survey (NHIS), conducted by the Centers for Disease Control and Prevention's (CDC) and National Center for Health Statistics (NCHS) till 2007, energy medicine was practiced almost 4 out of 10 adults in the past 12 months [ 35]. Current experiment was designed to demonstrate the impact of Mr. Trivedi's biofield treatment on K. pneumoniae for its antimicrobial susceptibility testing. Further, the molecular methods was performed to study the genetic alterations and similarities using RAPD sequencing methods. Increased infection of K. pneumoniae and other Gram-negative pathogen associated with nosocomial infections have become a global health problem. Results suggest that biofield treatment on microorganism can alter the in vitro sensitivity of the antimicrobials and it might be correlated with acetylation of antimicrobials that may happen via active drug efflux mechanism. Increased incidence of nosocomial infections and broad resistance against broad spectrum antibiotics would be a serious global threat. Mr. Trivedi's biofield treatment showed a significant decrease in MIC values of ampicillin/sulbactam and chloramphenicol along with improved sensitivity, which suggested an alteration at enzymatic/ genetic level that may modify ligand-receptor interaction. Hence a cascade of intra-cellular signals may be initiated, accelerated or inhibited [ 36], after biofield treatment on pathogenic microbes. Further, biofield treatment on K. pneumoniae , possible involve alterations at receptor level due to energy transfer via. biofield treatment, which may change the receptor drug interactions, which in turn alter the internal state of the microbe. However, it was reported that electromagnetic fields might alter the transmembrane concentration of cell, and will alter the receptor protein molecule. Biofield treatment might alter the receptor interaction, and results in altered antibiogram of K. pneumoniae with respect to the control [ 37].

Conclusion

In summary, it can be concluded that Mr. Trivedi's biofield energy treatment on K. pneumoniae has improved the antimicrobial sensitivity and decreased the MIC value by two folds in case ampicillin/sulbactam and chloramphenicol. Cephalothin showed altered antibiogram after biofield treatment as compared to the control. The biochemical study results showed the alteration of biochemical reactions of cephalothin, colistin, and nitrate, followed by change in the biotype number (7774 4272) after biofield treatment as compared to the control biotype (7774 4274). Using RAPD markers, the biofield treated samples were characterized and showed interspecific polymorphic relationship with K. pneumoniae after biofield treatment. Overall, it seems that Mr. Trivedi's unique biofield energy treatment on pathogenic microbes might be used as an alternate approach to alter the antimicrobial sensitivity.

Acknowledgement

This work was supported by Trivedi Science ™, Trivedi Master Wellness ™ and Trivedi Testimonials. Authors acknowledge the generosity and cooperation of all participating members from PD Hinduja National Hospital and MRC, Mumbai, Microbiology Lab for conduction antimicrobial studies. Authors are thankful to Bangalore Genei Private Limited, for conducting RAPD analysis.

References

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2. Chetoui H, Delhalle E, et al. (1999) Epidemiological typing of extended spectrum b-lactamase-producing Klebsiellapneumoniae isolates by pulsed-field gel electrophoresis and antibiotic susceptibility patterns. Res Microbiol 150: 265-272.

3. Vernet V, Philippon A, et al. (1995) Virulence factors (aerobactin and mucoid phenotype) in Klebsiellapneumoniae and E. coli blood culture isolates. FEMS Microbiol Lett 130: 51-57.

4. Corkill JE, Cuevas LE, Guegel RQ, Greensill J, Hart CA (2001) SHV-27, a novel cefotaxime-hydrolysing beta-lactamase, identified in Klebsiellapneumoniae isolates from a Brazilian hospital. J AntimicrobChemother 47: 463-465.

5. Trivedi MK, Patil S, Shettigar H, Bairwa K, Jana S (2015) Phenotypic and biotypic characterization of Klebsiellaoxytoca: An impact of biofield treatment. J Microb Biochem Technol 7: 203-206.

6. Rubik B, Brooks AJ, Schwartz GE (2006) In vitro effect of Reiki treatment on bacterial cultures: Role of experimental context and practitioner wellbeing. J Altern Complement Med 12: 7-13.

7. Chein CH, Tsuei JJ, Lee SC, Huang YC, Wei YH (1991) Effect of emitted bioenergy on biochemical function of cells. Am J Chin Med 19: 285-292.

8. Mo Z, Chen KW, Ou W, Li M (2003) Benefits of external qigong therapy on morphine-abstinent mice and rats. J Altern Complement Med 9: 827-835.

9. Movaffaghi Z, Hassanpoor M, Farsi M, Hooshmand P, Abrishami F (2006) Effects of therapeutic touch on blood hemoglobin and hematocrit level. J Holist Nurs 24: 41-48.

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13. Oschman JL (2000) Energy medicine: The scientific base. (2nd edn). Edinburgh: Churchill Livingston.

14. Trivedi MK, Tallapragada RR (2008) A transcendental to changing metal powder characteristics. Met Powder Rep 63: 22-28, 31.

15. 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.

16. 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 field treatment. IndEng Manage 4: 161.

17. Sances F, Flora E, Patil S, Spence A, Shinde V (2013) Impact of biofield treatment on ginseng and organic blueberry yield. Agrivita J AgricSci 35: 22-29.

18. Lenssen AW (2013) Biofield and fungicide seed treatment influences on soybean productivity, seed quality and weed community. Agricultural Journal 8: 138-143.

19. Trivedi MK, Patil S, Shettigar H, Gangwar M, Jana S (2015) An effect of biofield treatment on multidrug-resistant Burkholderiacepacia: A multihost pathogen. J Trop Dis 3: 167.

20. Trivedi MK, Patil S, Shettigar H, Gangwar M, Jana S (2015) Antimicrobial sensitivity pattern of Pseudomonas fluorescens after biofield treatment. J Infect Dis Ther 3: 222.

21. Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18: 6531-6535.

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24. Fader RC, Weaver E, et al. (2013) Multilaboratory study of the biomic automated well-reading instrument versus MicroScanWalkAway for reading MicroScan antimicrobial susceptibility and identification panels. J Clin Microbiol 51: 1548-1554.

25. Ashayeri-Panah M, Eftekhar F, Ghamsari MM, Parvin M, Feizabadi MM (2013) Genetic profiling of Klebsiellapneumoniae: Comparison of pulsed field gel electrophoresis and random amplified polymorphic DNA. Braz J Microbiol 44: 823-828.

26. Hoffman AM, Viel L, Muckle CA, Yager JS, Staempfli HR (1992) Evaluation of sulbactam plus ampicillin for treatment of experimentally induced Klebsiellapneumoniae lung infection in foals. Am J Vet Res 53: 1059-1067.

27. Haeggman S, Lofdahl S, Burman LG (1997) An allelic variant of the chromosomal gene for class A beta-lactamase K2, specific for Klebsiellapneumoniae, is the ancestor of SHV-1. Antimicrob Agents Chemother 41: 2705-2709.

28. Jones RN (2001) In vitro evaluation of ertapenem (MK-0826), a long-acting carbapenem, tested against selected resistant strains. J Chemother 13: 363-376.

29. Kumar AR (2013) Antimicrobial sensitivity pattern of Klebsiella pneumonia isolated from pus from tertiary care hospital and issues related to the rational selection of antimicrobials. J Chem Pharm Res 5: 326-331.

30. Jones RN, Baquero F, Privitera G, Inoue M, Wiedemann B (1997) Inducible ß-lactamase-mediated resistance to third-generation cephalosporins. Clin Microbiol Infect 3: s7-s20.

31. Hansen DS, Aucken HM, Abiola T, Podschun R (2004) Recommended test panel for differentiation of Klebsiella species on the basis of a trilateral interlaboratory evaluation of 18 biochemical tests. J Clin Microbiol 42: 3665-3669.

32. Eisen D, Russel EG, et al. (1995) Random amplified polymorphic DNA and plasmid analysis used in investigation of an outbreak of multiresistantKlebsiellapneumoniae. J Clin Microbiol 33: 713-717.

33. Williams JG, Hanafey MK, Rafalski JA, Tingey SV (1993) Genetic analysis using random amplified polymorphic DNA markers. Methods Enzymol 218: 704-740.

34. Orskov I, Orskov F (1984) Serotyping of Klebsiella. Methods in microbiology. NY: Academic Press, Inc. New York.

35. Barnes PM, Bloom B, Nahin RL (2008) Complementary and alternative medicine use among adults and children: United States, 2007. Natl Health Stat Report 10: 1-23.

36. Hintz KJ, Yount GL, et al. (2003) Bioenergy definitions and research guidelines. AlternTher Health Med 9: A13-A30.

37. Lindstrom E, Mild KH, Lundgren E (1998) Analysis of the T cell activation signaling pathway during ELF magnetic field exposure, p56lck and [Ca2+]i-measurements. Bioeletrochem Bioenerg 46: 129-137.

# data[].attributes.attachment_link.title
Antimicrobial Susceptibility, Biochemical Characterization and Molecular Typing of Biofield Treated Klebsiella pneumoniae

# data[].attributes.attachment_link.description
Mahendra Kumar Trivedi1 , Alice Branton1 , Dahryn Trivedi1 , Mayank Gangwar2 and Snehasis Jana2 * 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, Madhya Pradesh, India *Corresponding Author: Dr. Snehasis Jana Trivedi Science Research Laboratory Pvt.

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		An Experimental Study on Biofield Energy Treated p-Anisidine
	

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		An Experimental Study on Biofield Energy Treated p-Anisidine
	

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| Read this experimental study conducted by Mahendra Trivedi and the research team, to evaluate the impact of biofield energy treatment on the physicochemical and spectroscopic characterization of p-Anisidine.

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		Isotopic Abundance Analysis of Biofield Treated Benzene, Toluene and p-Xylene
	

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		Isotopic Abundance Analysis of Biofield Treated Benzene, Toluene and p-Xylene
	

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| Study the report to check and analyze the isotopic abundance ratio of biofield energy treated benzene, toluene and p-xylene using gas chromatography-mass spectrometry. Visit the link to know more in detail.

# data[].attributes.attachment_link.keywords[]
biofield treatment; benzene; gas chromatography-mass spectrometry; toluene; p-xylene

# data[].attributes.attachment_link.title
Isotopic Abundance Analysis of Biofield Treated Benzene, Toluene and p-Xylene Using Gas Chromatography-Mass Spectrometry (GC-MS)

# data[].attributes.attachment_link.description
Benzene, toluene and p-xylene are derivatives of benzene, generally produced from crude petroleum and have numerous applications in industry. The aim of the present study was to ev..

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		Biofield Energy Treated Manganese Sulfide Powder | eScholarship
	

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		Biofield Energy Treated Manganese Sulfide Powder | eScholarship
	

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| The present study was designed to evaluate the effect of Mr. Trivedi's biofield energy treatment on the atomic and physical properties of manganese sulfide powder using X-ray diffraction, Fourier transform infrared and electron spin resonance spectroscopy. Join the University of California network to know more about this research.

# data[].attributes.attachment_link.title
Characterization of Atomic and Physical Properties of Biofield Energy Treated Manganese Sulfide Powder

# data[].attributes.attachment_link.description
Manganese sulfide (MnS) is known for its wide applications in solar cell, opto-electronic devices, and photochemical industries. The present study was designed to evaluate the effect of biofield energy treatment on the atomic and physical properties of MnS. The MnS powder sample was equally divided into two parts, referred as to be control and to be treated.

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		Human Biofield Energy Treated Hi VegTM Acid Hydrolysate
	

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		Human Biofield Energy Treated Hi VegTM Acid Hydrolysate
	

# data[].attributes.attachment_link.title
Physicochemical Characterization of Biofield Energy Treated Hi VegTM Acid Hydrolysate

# data[].attributes.attachment_link.description
The hydrolysed vegetable proteins are acidic or enzymatic hydrolytic product of proteins derived from various sources such as milk, meat or vegetables. The current study was designed to evaluate the impact of biofield energy treatment on the various physicochemical and spectra properties of Hi VegTM acid hydrolysate i.e. a hydrolysed vegetable protein.

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		Spectroscopic Characterization of Yeast Extract Powder | Biofield Treatment
	

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		Spectroscopic Characterization of Yeast Extract Powder | Biofield Treatment
	

# data[].attributes.body
| Yeast extract powder is particularly used in culture media for the cultivation of microorganisms found in milk or other dairy products. The present study was intended to explore the influence of Mr. Trivedi's biofield energy treatment on the physicochemical and spectral properties of YE powder.

# data[].attributes.attachment_link.title
Physicochemical and Spectroscopic Characterization of Yeast Extract Powder After the Biofield Energy Treatment

# data[].attributes.attachment_link.description
Yeast extract powder (YE powder) is particularly used in culture media for the cultivation of microorganisms found in milk or other dairy products. The present study was intended to explore the influence of biofield energy treatment on the physicochemical and spectral properties of YE powder.

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		Phosphate Buffer Saline and Hanks Balanced Salt Medium | eScholarship
	

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		Phosphate Buffer Saline and Hanks Balanced Salt Medium | eScholarship
	

# data[].attributes.body
| The current research study was aimed to investigate the influence of Mr. Trivedi’s biofield energy treatment on the physicochemical properties of the Phosphate Buffer Saline and Hanks Balanced Salt Medium. Study the report to know more!

# data[].attributes.attachment_link.title
Comparative Physicochemical Evaluation of Biofield Treated Phosphate Buffer Saline and Hanks Balanced Salt Medium

# data[].attributes.attachment_link.description
Phosphate buffer saline (PBS) has numerous biological and pharmaceutical applications. Hank buffer salt (HBS) has been used as a medium for tissue culture applications. This research study was aimed to investigate the influence of Mr. Trivedi's biofield energy treatment on physicochemical properties of the PBS and HBS.

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		Human Biofield Energy Treated Alphonso Mango (Mangifera indica L.)
	

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		Human Biofield Energy Treated Alphonso Mango (Mangifera indica L.)
	

# data[].attributes.attachment_link.title
Morphological Characterization, Quality, Yield and DNA Fingerprinting of Biofield Energy Treated Alphonso Mango (Mangifera indica L.)

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Author(s): Mahendra, Trivedi; Trivedi Global Inc. | Abstract: Alphonso is the most delicious variety of mango (Mangifera indica L.) known for its excellent texture, taste, and richness with vitamins and minerals. The present study was attempted to evaluate the impact of Mr. Trivedi's biofield energy treatment on morphological characteristics, quality, yield and molecular assessment of mango.

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		Vegetative Growth Parameters in Biofield Treated Bottle Gourd and Okra
	

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		Vegetative Growth Parameters in Biofield Treated Bottle Gourd and Okra
	

# data[].attributes.attachment_link.title
Evaluation of Vegetative Growth Parameters in Biofield Treated Bottle Gourd (Lagenaria siceraria) and Okra (Abelmoschus esculentus)

# data[].attributes.attachment_link.description
The objective of the study was to assess the growth contributing characters of biofield treated bottle gourd (Lagenaria siceraria) and okra (Abelmoschus esculentus) seeds. The seeds of both crops were divided into two groups, one was kept aside and denoted as untreated, while the other group was subjected biofield energy treatment.

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		Biofield Energy Treated Ortho and Meta Toluic Acid Isomers
	

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		Biofield Energy Treated Ortho and Meta Toluic Acid Isomers
	

# data[].attributes.attachment_link.title
Quantitative Determination of Isotopic Abundance Ratio of 13C, 2H, and 18O in Biofield Energy Treated Ortho and Meta Toluic Acid Isomers

# data[].attributes.attachment_link.description
O-Toluic acid (OTA) and m-toluic acid (MTA) are two isomers of toluic acid that act as an important organic intermediates, mostly used in medicines and pesticides. The aim of the study was to evaluate the impact of biofield energy treatment on isotopic abundance ratios of 2H/1H, 13C/12C, (PM+1)/PM and 18O/16O, (PM+2)/PM, in toluic acid isomers using gas chromatography-mass spectrometry (GC-MS).

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		Antimony Tin Oxide Nanopowder after Biofield Energy Treatment
	

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		Antimony Tin Oxide Nanopowder after Biofield Energy Treatment
	

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The Potential Impact of Biofield Energy Treatment on the Atomic and Physical Properties of Antimony Tin Oxide Nanopowder

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Antimony tin oxide (ATO) is known for its high thermal conductivity, optical transmittance, and wide energy band gap, which makes it a promising material for the display devices, solar cells, and chemical sensor industries. The present study was undertaken to evaluate the effect of biofield energy treatment on the atomic and physical properties of ATO nanopowder.

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		Scientific Research Validating The Effect of Biofield Energy on Fish Peptone
	

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		Scientific Research Validating The Effect of Biofield Energy on Fish Peptone
	

# data[].attributes.attachment_link.title
Physical, Spectroscopic and Thermal Characterization of Biofield Treated Fish Peptone

# data[].attributes.attachment_link.description
The by-products of industrially processed fish are enzymatically converted into fish protein isolates and hydrolysates having a wide biological activity and nutritional properties. However, the heat processing may cause their thermal denaturation thereby causing the conformational changes in them.

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		Spectroscopic Properties of Biofield Energy Treated Bio Peptone
	

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		Spectroscopic Properties of Biofield Energy Treated Bio Peptone
	

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| In the present research, the bio peptone sample was divided into two parts: control and (biofield) treated. The control sample was kept without treatment, while the treated sample was handed over in sealed pack to Mr. Trivedi for the biofield energy treatment. Follow the link to see and analyze the interesting result.

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Characterization of Physicochemical and Spectroscopic Properties of Biofield Energy Treated Bio Peptone

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Bio peptone is a combination of enzymatic digest of animal tissues and casein; and generally used for the growth of several varieties of microbes. The aim of present study was to investigate the impact of biofield energy treatment on the physicochemical and spectroscopic properties of bio peptone.

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		Biofield Effect on Butylatedhydroxytoluene and 4-methoxyphenol
	

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		Biofield Effect on Butylatedhydroxytoluene and 4-methoxyphenol
	

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| In the present research paper it is shown that the increased isotopic abundance ratio after Biofield treatment on phenol derivatives may increase the bond stability, which could result in reduce autocatalytic oxidation reactions.

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Trivedi Effect | Investigation of Isotopic Abundance Ratio of Biofield Treated Phenol Derivatives Using Gas Chromatography-Mass Spectrometry

# data[].attributes.attachment_link.description
Butylatedhydroxytoluene (BHT) and 4-methoxyphenol (4-MP) are phenol derivatives that are generally known for their antioxidant properties and depigmenting activities. The aim of this study was to evaluate the impact of biofield energy treatment on

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		Alteration in Characteristics of Escherichia Coli after Biofield Treatment
	

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		Alteration in Characteristics of Escherichia Coli after Biofield Treatment
	

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| Based on the experimental results, it is assumed Mr. Trivedi’s Biofield treatment could be applied to alter the sensitivity pattern of antimicrobials, against multi-drug resistance isolates of Escherichia coli.

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		Biofield Treatment | An Experimental Study on Phenol Derivatives
	

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		Biofield Treatment | An Experimental Study on Phenol Derivatives
	

# data[].attributes.body
| In this experimental study, butylatedhydroxytoluene and 4-methoxyphenol were studied under the influence of Biofield energy treatment and observed significant changes in isotopic abundance compared to the control sample.

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		Antimicrobial Susceptibility Assay of Biofield Treated Escherichia Coli
	

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		Antimicrobial Susceptibility Assay of Biofield Treated Escherichia Coli
	

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| The present study describes that there is percentage change in antimicrobial sensitivity pattern, minimum inhibitory concentrations and biochemical reactions after Biofield treatment in multi-drug resistant lab isolates of Escherichia coli.

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		Isotopic Abundance Ratio of Biofield Treated Phenol Derivatives
	

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		Isotopic Abundance Ratio of Biofield Treated Phenol Derivatives
	

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| This research paper reveals that antioxidant properties of phenol derivatives could be enhanced by Mr. Mahendra Trivedi’s unique Biofield energy treatment. Follow the link to know more about this research

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		Effect of Biofield Treatment on MDR Isolates of Escherichia Coli
	

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		Effect of Biofield Treatment on MDR Isolates of Escherichia Coli
	

# data[].attributes.body
| In this research MDR isolates of escherichia coli showed altered pattern of antimicrobial sensitivity as compared to its control sample in all the isolates after Biofield treatment. Visit the link to know more about this research

# data[].attributes.attachment_link.title
Investigation of Biofield Treatment on Antimicrobial Susceptibility, Biochemical Reaction Pattern and Biotyping of Enteropathogenic Multidrug-Resistant Escherichia coli Isolates

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PubFacts seeks to make the world's scientific research easy to locate, access, and collaborate on.

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		Biofield Impact on Characteristics of Pogostemon cablin
	

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		Biofield Impact on Characteristics of Pogostemon cablin
	

# data[].attributes.body
| The study revealed that a single spell of biofield energy treatment produced significant increase in growth in treated group throughout all the morphogenetic phases from in vitro to in vivo level.

# data[].attributes.attachment_link.title
Impact of Biofield Treatment on Growth and Anatomical Characteristics of Pogostemon cablin (Benth.)

# data[].attributes.attachment_link.description
Pogostemon cablin is a known aromatic plant which is cultivated for its essential oil widely applicated in perfumery and cosmetic industries. In the present study, the effect of biofield treatment was studied on the growth of P. cablin.

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		Biofield | Properties of Bile Salt and Proteose Peptone
	

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		Biofield | Properties of Bile Salt and Proteose Peptone
	

# data[].attributes.body
| The present study has evaluated the influence of biofield treatment on thermal and physical properties of Bile Salt and Proteose Peptone.

# data[].attributes.attachment_link.title
Peer Evaluation : Thermal and Physical Properties of Biofield Treated Bile Salt and Proteose Peptone

# data[].attributes.attachment_link.description
Abstract : Bile salt (BS) and proteose peptone (PP) are important biomacromolecules being produced inside the human body. The objective of this study was to investigate the influence of biofield treatment on physicochemical properties of BS and PP. The study was performed in two groups (control and treated).

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		Biofield Treatment Effect in a Model of Cancer Cell Death
	

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		Biofield Treatment Effect in a Model of Cancer Cell Death
	

# data[].attributes.body
| This study assessed the potential influence of biofield treatment on cultured human cancer cells and whether such influence was affected by varying the duration of the treatment (dose) or the distance.

# data[].attributes.attachment_link.readable_text
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The snippet could not be located in the article text. This may be because the snippet appears in a figure legend, contains special characters or spans different sections of the article.

Journal of Alternative and Complementary Medicine

J Altern Complement Med. 2013 February; 19(2): 124-127.

Garret Yount, PhD, ¹ Shrikant Patil, BDS, ² Umang Dave, BS, ¹ Leonardo Alves-dos-Santos, BS, ¹ Kimberly Gon, BS, ¹ Robert Arauz, BS, ¹ and Kenneth Rachlin, MS ¹

Abstract

Objective

This study assessed the potential influence of biofield treatment on cultured human cancer cells and whether such influence was affected by varying the duration of the treatment (dose) or the distance between the biofield practitioner and the target cells.

Design

Biofield treatment dosage was assessed from a short distance (0.25 meters) in three independent experiments involving 1, 2, or 5 treatments, along with another set of three independent and comparable mock experiments. Biofield treatment distance was assessed at 0.25, 25, and ~ 2000 meters involving two treatments in three independent experiments along with another set of three mock experiments.

Intervention

Biofield treatments were delivered by a highly acclaimed biofield practitioner with the intention of diminishing growth of the cells or inducing cancer-cell death.

Outcome measure

Cell viability was quantified 20 hours after treatments, using a spectrophotometric assay for live-cell counting. The dependent measure for each experiment was the log ratio of the cell viability values of treated samples (biofield or mock) over the values of untreated control samples.

Results

A trend of decreasing cell viability with increasing biofield dose was evident in the first set of experiments assessing dose-response; however, no such effect was evident in the second set of experiments evaluating biofield treatment distance. Mock experiments yielded relatively stable viability ratios in both sets of experiments. Linear regression analysis and hypothesis testing of the data taken as a whole did not yield statistical significance at p<0.05.

Conclusions

These results represent the first indication of a biofield treatment dose-response in a controlled laboratory setting. The data are inconclusive because of the inability of reproduce the cellular response in a replicate experiment.

Introduction

Within the burgeoning field of complementary and alternative medicine, the domain of biofield treatments, or energy medicine, is replete with both faith and skepticism. Biofield treatments involve a putative bioenergy that is intentionally channeled by a practitioner to heal patients from a distance. A common assumption among biofield practitioners is that increased dosage, typically in the form of repeated treatments-and even increased distance over which the treatments are delivered-can result in increased efficacy. This trial sought to examine this assumption, using human cancer cells cultured in vitro as the target of biofield treatments. Two sets of experiments were conducted under strict laboratory conditions: one evaluating varying dosage and a second evaluating biofield treatment delivered from varying distances.

Materials and Methods

Cell culture and handling

Human U87 glioblastoma cells were expanded in standard culture media (RPMI-1640, Sigma-Aldrich, St. Louis, MO) supplemented with 10% fetal bovine serum and divided into aliquots for storage in liquid nitrogen. At the start of each experiment, an aliquot was thawed and cells were seeded into four 96-well culture plates at 1000 cells/well. The plates were each labeled with an identifying code and placed in a humidified incubator (designated as incubator #1) set at 37°C and 5% CO ₂. After 4 hours of incubation, two of the plates were selected randomly by a second scientist to receive treatment (biofield or mock), transferred to a treatment room, and placed in a humidified incubator with clear walls (incubator #2). The remaining two plates served as controls and received no treatment but were taken out of the incubator for 2 minutes at the beginning and end of the treatment period to mimic the change in ambient temperature that occurred with the treatment plates during their transfer. At the completion of the treatment period, the two treatment plates were returned to incubator #1. All four plates remained undisturbed overnight and cell viability was assessed the following day.

Cell viability analysis

Cell viability was quantified 20 hours after the initiation of experimental treatments (biofield or mock) using the Cell Count Kit-8 (Dojindo Laboratories, Santa Clara, CA), a colorimetric assay based on a tetrazolium salt that is reduced by dehydrogenase activities within cells to produce a yellow-color formazan dye. The amount of the formazan dye generated in the cell-culture media is directly proportional to the number of living cells. The absorbance was read, using a spectrophotometer with a test wavelength of 450 nm. Data were acquired from a total of 30 independent wells in each plate, avoiding wells on the edge of the culture plate. Background absorbance was compensated for, using measurements obtained from six wells that contained only media (no cells). The average absorbance of these wells was subtracted from the other wells.

Because of natural variation in cell-viability measurement from day to day, the dependent measure for each experiment was the ratio of the cancer-cell viability values from treated cultures (biofield or mock) over the values from control cultures. Cell viability ratios were computed by dividing the mean values from the duplicate plates kept in incubator #2 during the treatment period (a total of 60 wells) by the mean values from the duplicate plates in the control incubator (a total of 60 wells). Because ratios are inherently asymmetrical, a log transform was applied to the ratios. A log ratio value less than zero indicates cancer cell death or diminished growth following treatment compared to the control condition and a value greater than zero indicates the opposite.

Biofield and mock treatments

All biofield treatments were delivered by one practitioner who is recognized internationally as a healer and who has developed a technique of mental energy transmission based on innate healing abilities. Following published guidelines, the practitioner remains anonymous to avoid any potential conflict of interest. Each treatment lasted for 5 minutes. For increased dosage, treatments were repeated with 10-minute rest periods. During treatments from a close distance, the practitioner was escorted into the treatment room and seated in front of incubator #2. Distant treatments were delivered remotely, aided by a webcam link to a live image of the cells in incubator #2. During mock treatments, the duplicate plates were placed in incubator #2, and the room remained empty for the treatment time without the practitioner being involved.

Systematic negative controls

The experiments involving mock treatments served as systematic negative controls throughout the study; that is, each set of experiments included an equal number of experiments in which all of the physical manipulations of the cells and subsequent analyses were carried out except for the experimental intervention. This method of assessing variability intrinsic to the experimental system was considered applicable based on its usefulness in the study of nonionizing electromagnetic fields.

Blinding and randomization

The experimental protocol was divided between two scientists to insure that the scientist handling the cells and analyzing the data was not aware of whether the cells had received biofield treatment, following published guidelines. Identifying codes and group assignments were determined using the random number generator function of a scientific calculator.

Results

Six independent experiments were conducted to evaluate the potential for varying biofield treatment dosage: three involving increasing numbers of biofield treatments (1, 2, and 5 treatments with a total cumulative duration of 5, 10, and 25 minutes, respectively); and three involving only mock treatments. The cell-viability ratios for the three mock/control experiments were all close to zero while those involving biofield treatments of increasing dosage appeared to be monotonically decreasing (see ). Thus, the greatest cancer-cell inhibition was observed when the practitioner was closest and delivered the highest dose.

Viability plots for cancer cells exposed to either mock or biofield treatments. Cell viability ratios are plotted for (A) a set of experiments evaluating biofield dosage and (B) biofield treatment distance. Points represent the log-transform of the ratios ...

A second set of six experiments was conducted: three involving the same biofield treatment dosage (2 treatments with a total cumulative duration of 10 minutes) from various distances (0.25, 25, and ~ 2000 meters); and three involving mock treatments. None of the resulting cell-viability ratios in this second set of experiments were significantly different than zero when considering their standard deviations (see ). Notably, the experiment involving the treatment from the closest distance in these experiments is a replication of the experiment in the first set of experiments involving the 10-minute biofield treatment but the results were in opposite directions.

A weighted linear regression was performed to assess the monotonically decreasing dose-response effect using the model: Y=B ₀+B ₁X, for which X is biofield treatment time, Y is log ratio of cell viability, and the transformed standard deviations are used as weights. The data point from the replicate experiment in the second set of experiments (treatment from closest distance) was included. Although the estimated slope (B ₁) was −0.039 (and this is consistent with a trend of decreasing cell viability as a function of treatment duration) the parameter estimation was not well-determined ( p=0.348). Linear hypothesis testing against the null hypothesis (Y=0), using the estimated parameters and covariance from the linear regression analysis did not yield statistical significance ( p=0.410). Nonparametric hypothesis testing of the data considered as a whole (six biofield/control ratios versus six mock/control ratios) did not yield significance via the Wilcoxon rank-sum test ( p=0.699) or via the Kruskal-Wallis test ( p=0.631). summarizes the underlying data.

Cell Viability Values and Subsequent Calculations Underlying the Ratio Values for All 12 Independent Experiments

Discussion

The apparent dose-response observed in the first set of experiments is provocative, particularly in light of the results of the systematic negative control experiments conducted in parallel that indicated a relatively stable baseline variability in the data output from the model system. However, the interpretation of these results is complicated by the lack of replicability observed in the second set of experiments using the same model system and the same participants. Part of the struggle is reconciling the centuries of precedent in which the test of replicability has been a fundamental tenet of the scientific method, and the acknowledgment that the experimental intervention in this model system derives from a human being and may thus be inherently changing with time and situation (e.g., mental and emotional state at a particular time). Lack of replicability has been a consistent element in a series of in vitro studies of various biofield treatment modalities conducted by the current authors' group and seems to be consistent with the extant data across the field. For example, the lack of independent replications was noted as an important issue in critical reviews that assessed in vitro studies of external qigong, another form of biofield treatment. In vitro models can be advantageous in terms of minimizing confounding variables; yet, these models may be suboptimal for investigating biofield treatment efficacy for a number of reasons. Just one example is that biofield therapies may require the presence of an organized biofield in the target system for optimal effect, and this element would be disrupted or dissipated in cell cultures separated from the body. While there are encouraging data indicating that some disadvantages of in vitro models might be overcome through experimental design considerations specific to biofield treatment models, the current authors believe that in vivo models are more appropriate for investigating biofield treatments at the current stage of this research where resources are limited and evidence of efficacy remains controversial. Questions regarding the underlying mechanisms of action may ultimately be addressed in vitro better, but in vivo models offer the most hope for evaluating the efficacy of biofield treatments and would thus serve the most urgent public health needs better.

Conclusions

The results of this trial represent the first indication of a biofield treatment dose-response in a controlled laboratory setting. The data are inconclusive because of the inability to reproduce the cellular response in a replicate experiment.

Acknowledgments

The authors wish to thank Tri Luu, AS, for technical assistance. The work was supported by a grant from the Trivedi Foundation.

Disclosure Statement

No competing financial interests exist.

References

1. Lu ZY. Scientific Qigong Exploration: The Wonders and Mysteries of Qi. Malvern, PA: Amber Leaf Press; 1997. p. 171.

3. Walleczek J. Shiu E,C. Hahn GM. Increase in radiation-induced HPRT gene mutation frequency after nonthermal exposure to nonionizing 60[ Hz electromagnetic fields. Radiat Res. 1999;151:489-497. PubMed]

6. Taft R. Moore D. Yount G. Time-lapse analysis of potential cellular responsiveness to Johrei, a Japanese healing technique. BMC Complement Altern Med. 2005;24(5):2. [ PMC free article]

7. Yount G. Solfvin J. Moore D, et al. In vitro test of external qigong. BMC Complement Altern Med. 2004;15(4):5. [ PMC free article]

Articles from Journal of Alternative and Complementary Medicine are provided here courtesy of Mary Ann Liebert, Inc.

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Effect of Biofield Treated Energized Water on the Growth and Health Status in Chicken (Gallus gallus domesticus)

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 Road, Bhopal- 462026, Madhya Pradesh, India

*Corresponding Author:

Snehasis Jana
Trivedi Science Research Laboratory Pvt. Ltd.
Hall-A, Chinar Mega Mall, Chinar Fortune City
Hoshangabad Road, Bhopal- 462026, Madhya Pradesh, India
Tel: +91-755-6660006
E-mail: [email protected]

Received Date: September 08, 2015; Accepted Date: September 28, 2015; Published Date: September 30, 2015

Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Mondal SC, (2015) Effect of Biofield Treated Energized Water on the Growth and Health Status in Chicken ( Gallus gallus domesticus). Poult Fish Wildl Sci 3:140.doi:10.4172/2375-446X.1000140

Copyright: © 2015 Jana S, 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.

The current study was attempted to investigate the effect of Mr. Trivedi's biofield energy treated energized water on chicken. The total 4200 chicks were equally divided into two groups i.e. control and treated. The biofield treated energized water was provided to the treated chicks, while the control chicks were drunk with standard drinking water. During the experiment the parameters such as mortality, body weight, food intake etc. were assessed in both control and energized water treated birds. The mortality rate was reduced in the energized water treated chicks as 54.55% in week 1, 42.11% in week 6, and 39.13% in week 4, as compared to the control chicks. Moreover, the average body weight was increased by 12.50% in week 1 as compared to the control chicks. The feed conversion ratio was gradually decreased which indicated that the energized water treated chicks took less feeds while the body weight was increased in comparison to the control chicks. Besides, the energized water treated birds showed statistically significant (p<0.007) with 15.47% increase in the edible meat weight as compared to the control chicks. Moreover, the feather, skin and internal organ weight were significantly reduced by 21.22% (p<0.001) of energized water treated chicks as compared to the untreated chicks. The protein content was increased by 10.11% and cholesterol was decreased by 4.64% in birds of the treated group as compared to the control. The European efficiency factor was also increased by 10.67% in the energized water treated birds as compared to the control chicks.. The European efficiency factor was also increased by 10.67% in the energized water treated birds as compared to the control chicks. Altogether, the results suggest that Mr. Trivedi's biofield treated energized water could be a cost effective feeding approach in chicken production.

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		Biofield Energy Treatment Impact on Eggplant and Watermelon Crops
	

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| The present study was designed to evaluate the impact of biofield energy treatment on selected plots (control and treated) for the seeds of eggplant and watermelon crop. Genetic variability parameters of both the crops were studied using RAPD (DNA fingerprintign).

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		Analyze Eggplant and Watermelon Crops after an Impact of Biofield Energy
	

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		Analyze Eggplant and Watermelon Crops after an Impact of Biofield Energy
	

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| The study to check and analyze the human biofield energy influence on the growth of the overall crop yield of eggplant and watermelon. Read this report in detail to know more!

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		Human Biofield Energy Influence on Eggplant and Watermelon Crops
	

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		Human Biofield Energy Influence on Eggplant and Watermelon Crops
	

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| Visit & know the impact of human biofield energy influence on the growth of Eggplant (Solanum melongena L.) and Watermelon crops (Citrullus lanatus), a test by Mahendra Kumar Trivedi and the research team.

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		Eggplant and Watermelon Crops after Biofield Treatment
	

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		Eggplant and Watermelon Crops after Biofield Treatment
	

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| The study conducted by Mahendra Trivedi and the research team, to analyze the overall growth of the two inbreed crop varieties after the biofield energy treatment. The plots were selected for the study, and divided into two parts, control and treated.

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		Journal | Advances in Crop Science and Technology | Biofield
	

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		Journal | Advances in Crop Science and Technology | Biofield
	

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| The eggplant (Solanum melongena L.) is considered as one of the most important fruit vegetable crops all over the World. Watermelon (Citrullus lanatus) belongs to the family of Cucurbitaceae, grown as a cash crop.

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		Evaluate Growth of Eggplant and Watermelon Crops after Biofield Treatment
	

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		Evaluate Growth of Eggplant and Watermelon Crops after Biofield Treatment
	

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| Research done by Mahendra Kumar Trivedi – Study the report to evaluate the growth of the Eggplant and Watermelon crops after the impact of biofield energy treatment on it.

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		Mahendra Trivedi's Biofield Treated Eggplant and Watermelon Crops
	

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		Mahendra Trivedi's Biofield Treated Eggplant and Watermelon Crops
	

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| The study data concluded that Mr. Trivedi’s biofield energy treatment has the ability to alter the plant growth rate, and can be utilized in better way as compared with the existing agricultural crop improvement techniques to improve the overall crop yield.

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		Biofield Treatment's Effect on Eggplant and Watermelon Crops
	

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		Biofield Treatment's Effect on Eggplant and Watermelon Crops
	

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| The eggplant (Solanum melongena L.) and Watermelon (Citrullus lanatus), as one of the important vegetative crops have grown worldwide. The aim of the current study was to analyze the overall growth of the two inbreed crops varieties after the biofield treatment.

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		An Impact of Biofield Energy on Eggplant and Watermelon Crops
	

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		An Impact of Biofield Energy on Eggplant and Watermelon Crops
	

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|  Eggplant and watermelon, as one of the important vegetative crops have grown worldwide. Study the report conducted by Mr. Mahendra Trivedi to analyze the overall growth of the two inbreed crops varieties after biofield energy treatment.

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		Molecular Analysis of Eggplant and Watermelon Crops | Biofield Energy
	

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		Molecular Analysis of Eggplant and Watermelon Crops | Biofield Energy
	

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| Read this an open access research article of Mr. Mahendra Trivedi published in esciencecentral.org. The study data summarized that biofield energy treatment on the eggplant (Solanum melongena L.) and watermelon showed improved growth characteristics such as fruits, leaves and free from pest attack.

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		Biofield | Journal of Environmental Analytical Chemistry
	

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		Biofield | Journal of Environmental Analytical Chemistry
	

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| Read this research article conducted by Mr. Mahendra Trivedi and the research team to investigate the physical, thermal, and spectroscopic characterization of biofield energy treated Methyl-2-Naphthyl Ether. Follow the link to know more!

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		Spectroscopic Characterization of p-Nitroaniline | eScholarship
	

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		Spectroscopic Characterization of p-Nitroaniline | eScholarship
	

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| Para nitroaniline is an organic compound, used as an intermediate in the synthesis of pharmaceutical drugs, gasoline and dyes. Study the report to assess the impact of human biofield treatment on the spectroscopic and physicochemical properties of p-Nitroaniline.

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		Biofield Treatment Effect on Citrobacter braakii: A Urinary Pathogen
	

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		Biofield Treatment Effect on Citrobacter braakii: A Urinary Pathogen
	

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| Citrobacter braakii (C. braakii) is widespread in nature, mainly found in human urinary tract. Learn about the study conducted by Mahendra Trivedi to evaluate the phenotyping and 16s rDNA analysis after biofield energy treatment on the C. braakii.

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		Staphylococcus aureus after Biofield Treatment | eScholarship
	

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		Staphylococcus aureus after Biofield Treatment | eScholarship
	

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| Staphylococcus aureus is the key organism for food poisoning due to massive production of heat stable exotoxins. The study data suggested that biofield treatment has a significant impact on S. aureus in lyophilized as well as revived state. Follow the link to read more!

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		Nocardia otitidis | Biofield Treatment | eScholarship
	

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		Nocardia otitidis | Biofield Treatment | eScholarship
	

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| Nocardiosis is a soil-borne aerobic infection caused by Nocardia species commonly affects the respiratory tract. N. otitidis are the key organism for non-mycobacterial tuberculosis. Study this report to evaluate the antibiogram, genotype and phylogenetic analysis of biofield energy treated Nocardia otitidis.

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		Biofield Energy Treated Multidrug Resistant Strains of Staphylococcus Species
	

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		Biofield Energy Treated Multidrug Resistant Strains of Staphylococcus Species
	

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| The current study was designed to evaluate the Mahendra Kumar Trivedi’s biofield energy treatment impact on multidrug resistant clinical lab isolates of Staphylococcus species viz. Staphylococcus haemolyticus, Staphylococcus epidermidis and Staphylococcus aureus.

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		Biofield | Journal of Pharmaceutical Care & Health Systems
	

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		Biofield | Journal of Pharmaceutical Care & Health Systems
	

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| The study conducted by Mahendra Kumar Trivedi to assess the antibiogram of multidrug-resistant isolates of Enterobacter aerogenes after biofield energy treatment. Visit the link to know more about this research article in detail.

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		Study Klebsiella pneumoniae after Biofield Treatment | eScholarship
	

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		Study Klebsiella pneumoniae after Biofield Treatment | eScholarship
	

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| Study this an open access research article of Mr. Mahendra Trivedi and analyze the antimicrobial susceptibility, biochemical characterization and molecular typing of biofield energy treated Klebsiella pneumoniae. Follow the link to read more.

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		Biofield Energy Treatment Impact on p-Anisidine | eScholarship
	

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		Biofield Energy Treatment Impact on p-Anisidine | eScholarship
	

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| The p-anisidine is widely used as chemical intermediate in the production of various dyes, pigments, and pharmaceuticals. The study to evaluate the physicochemical and spectroscopic characterization of biofield energy treated p-Anisidine.

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		Analyze Biofield Treatment Effect on Benzene, Toluene and p-Xylene
	

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		Analyze Biofield Treatment Effect on Benzene, Toluene and p-Xylene
	

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| A test conducted by Mahendra Kumar Trivedi and the research team to check and analyze the isotopic abundance ratio of the biofield energy treated benzene, toluene and p-xylene using Gas Chromatography-Mass Spectrometry

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		p-Hydroxyacetophenone | Biofield | eScholarship
	

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		p-Hydroxyacetophenone | Biofield | eScholarship
	

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| P-Hydroxyacetophenone (pHAP) is an aromatic ketone derivative that is mainly used in the manufacturing of various pharmaceuticals, flavors, fragrances, etc. The study to investigate the effect of biofield energy treatment to improve the properties of  pHAP.

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		Biofield Treatment on Casein Enzyme Hydrolysate and Casein Yeast Peptone
	

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		Biofield Treatment on Casein Enzyme Hydrolysate and Casein Yeast Peptone
	

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| The study conducted by Mahendra Kumar Trivedi and the research team to evaluate the impact of biofield energy treatment on the physical and thermal properties of casein enzyme hydrolysate and casein yeast peptone.

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		Biofield | Improve the Quality of Beef Extract and Meat Infusion Powder
	

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		Biofield | Improve the Quality of Beef Extract and Meat Infusion Powder
	

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| The study to check the influence of Mahendra Trivedi's biofield treatment on two common flavoring agents used in food industries, namely beef extract powder (BEP) and meat infusion powder (MIP). Follow the link to read more.

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		Biofield Energy Treatment Effect on Neopentyl glycol | eScholarship
	

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		Biofield Energy Treatment Effect on Neopentyl glycol | eScholarship
	

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| Neopentyl glycol (NPG) has been extensively used as solid-solid phase change materials (PCMs) for thermal energy storage applications. A study was performed to evaluate the impact of biofield energy treatment on the physical, spectral and thermal properties of NPG. Read more

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		Disulfiram and Nicotinic Acid after Biofield Treatment | eScholarship
	

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		Disulfiram and Nicotinic Acid after Biofield Treatment | eScholarship
	

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| The study was designed to investigate the impact of Mr. Mahendra Kumar Trivedi  biofield energy treatment on spectral properties of disulfiram and nicotinic acid. Visit the link to know more about this research article in detail.

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		Characterization of Human Biofield Energy Treated Klebsiella oxytoca
	

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		Characterization of Human Biofield Energy Treated Klebsiella oxytoca
	

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| The current report depicted that Biofield treatment has the ability to make some alteration at the enzymatic, metabolic pathway and at genetic level of Klebsiella Oxytoca. Read this research to know more about the impact of Biofield treatment.

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		Impact of Human Biofield Energy Therapy on Klebsiella Oxytoca
	

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		Impact of Human Biofield Energy Therapy on Klebsiella Oxytoca
	

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| The present work was undertaken to evaluate the impact of Biofield energy treatment on  the antimicrobials susceptibility, biochemical reactions pattern and biotype of Klebsiella oxytoca.

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		Effect of Biofield Energy Treatment on the Inorganic Salts
	

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		Effect of Biofield Energy Treatment on the Inorganic Salts
	

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| The present study was attempted to investigate the impact of Biofield treatment on disodium hydrogen orthophosphate and sodium nitrate at atomic level like force constant, dipole moment and energy gap.

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		Characteristics of Klebsiella Oxytoca after  Biofield Energy Treatment
	

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		Characteristics of Klebsiella Oxytoca after  Biofield Energy Treatment
	

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| Klebsiella oxytoca (K. oxytoca) is a gram-negative microbe which provides a resistance against host defense mechanisms. Due to its clinical significance, a test was conducted to evaluate the effect of Biofield Treatment on phenotype and biotype characteristics of K. oxytoca.

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		Human Biofield Treated Disodium Hydrogen Orthophosphate and Sodium Nitrate
	

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		Human Biofield Treated Disodium Hydrogen Orthophosphate and Sodium Nitrate
	

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| Mr. Mahendra Trivedi and his research team conducted a test to investigate the impact of Biofield energy on the spectral properties of disodium hydrogen orthophosphate and sodium nitrate .

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		Biofield Energy Treatment Impact on Manganese Sulfide Powder
	

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		Biofield Energy Treatment Impact on Manganese Sulfide Powder
	

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| A test to investigate the impact of biofield energy treatment on the atomic and physical properties of manganese sulfide powder. To know more about biofield visit https://goo.gl/JJ1c3M

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		Biofield | Physical, Atomic and Thermal Properties of Lithium Powder
	

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		Biofield | Physical, Atomic and Thermal Properties of Lithium Powder
	

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| Learn the study and know more about the impact of biofield treatment on physical, atomic, and thermal properties of lithium powder.

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		An External (Biofield) Energy Effect on Bismuth Oxide Powder
	

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		An External (Biofield) Energy Effect on Bismuth Oxide Powder
	

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| Study to evaluate the effect of an external energy or biofield energy on the atomic, physical, and thermal properties of bismuth oxide powder.

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		Biofield Treatment on Characteristics of Selenium Powder
	

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		Biofield Treatment on Characteristics of Selenium Powder
	

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| The study conducted by Mahendra Trivedi, to evaluate the impact of biofield energy treatment on physical and thermal characteristics of selenium powder.

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		Biofield Energy Treated Barium Calcium Tungsten Oxide
	

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		Biofield Energy Treated Barium Calcium Tungsten Oxide
	

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| The study to check the the physical and structural properties of Mahendra Trivedi's biofield energy treated barium calcium tungsten oxide. To know more about biofield visit https://goo.gl/JJ1c3M

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		 Influence of Human Biofield Energy on Molybdenum Dioxide
	

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		 Influence of Human Biofield Energy on Molybdenum Dioxide
	

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| The study to check and analyze the influence of human biofield energy treatment on physical, thermal, and structural properties of molybdenum dioxide.

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	 Biofield | Physical & Thermal Properties of Silver Oxide
	

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	 Biofield | Physical & Thermal Properties of Silver Oxide
	

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| The biofield energy treatment impact on physical and thermal properties of Siver Oxide. The silver oxide XRD, TGA and other analysis shows the changes in properties of biofield treated samples as compared to the untreated samples.