Characterization of Antimicrobial Susceptibility Profile of Biofield Treated Multidrug-resistant Klebsiella oxytoca

Journal: Applied Microbiology: Open Access PDF  

Published: 06-Oct-15 Volume: 1 Issue: 1

DOI: 10.4172/2471-9315.1000101 ISSN: 2471-9315

Authors: Mahendra Kumar Trivedi, Alice Branton, Dahryn Trivedi, Gopal Nayak, Harish Shettigar, Mayank Gangwar and Snehasis Jana

Citation: Trivedi MK, Branton A, Trivedi D, Nayak G, Shettigar H, et al. (2016) Characterization of Antimicrobial Susceptibility Profile of Biofield Treated Multidrug-resistant Klebsiella oxytoca . Appli Micro Open Access 1: 1000101. doi:10.4172/2471-9315.1000101

  • 2703
  • 953

Abstract

Klebsiella are opportunistic pathogens that cause a wide spectrum of severe diseases. The aim of the present study was to investigate the impact of biofield treatment on multidrug resistant strain of K. oxytoca with respect to antibiogram pattern along with biochemical study and biotype number. Clinical lab isolate of K. oxytoca was divided into two groups i.e. control and treated. Control group remain untreated and treated group was subjected to Mr. Trivedi’s biofield. The analysis was done on day 10 after biofield treatment and compared with control group. Control and treated groups were analyzed for antimicrobial susceptibility pattern, minimum inhibitory concentration (MIC), biochemical reactions and biotype number using MicroScan Walk-Away® automated system. Experimental results showed the impact of biofield treatment on K. oxytoca and found alteration in both antimicrobial sensitivity and MIC values as compared with untreated group. Antimicrobial sensitivity of about 26.67% tested antimicrobials out of thirty was altered with respect to control. MIC results showed about 12.50% alterations in tested antimicrobials as compared to control. Biochemical study showed 24.24% alteration in tested biochemical reactions after biofield treatment. A significant change in biotype number (7713 5272) was identified after biofield treatment as compared to control (7775 4332). In treated group, a new species was identified as Kluyvera ascorbata, as compared to control, K. oxytoca. Study findings suggest that biofield treatment has a significant effect in altering the antimicrobial sensitivity, MIC values, biochemical reactions and biotype number of multidrug resistant strain of K. oxytoca. Biofield treatment could be applied to alter the antibiogram-resistogram pattern of antimicrobials.

Keywords:

Klebsiella oxytoca; Multidrug resistant; Antibiogram; Biofield treatment; Biochemical reactions; Biotyping

Introduction

Klebsiella oxytoca (K. oxytoca) is a Gram-negative pathogen, cylindrical rod shaped, non-motile in nature, and belongs to Enterobacteriaceae family. Klebsiella spp. are ubiquitous in environment [1], but K. oxytoca can be cultured from intestines of healthy humans and animals, oropharynx, mucous membrane and skin. K. oxytoca initially named as Aerobacter aerogens, which was identified as Klebsiella pneumoniae, but recent report classified it as K. oxytoca, on the basis of indole-positive test and ability to grow on melezitose, not in 3-hydroxybutyrate [2]. It is considered as an opportunistic pathogen, as most of the cases K. oxytoca-infected persons remain asymptomatic. However, K. oxytoca is now recognized as important clinical pathogen in hospitalized patients causing major nosocomial infections in children and neonates [3]. It is reported in many etiological human infections such as urinary tract infection, septic arthritis, bacteremia, septicemia, cholecystitis, soft tissue infections, and most recently in colicky neonates [1, 4-7]. During last few years, incidence of extended spectrum β-lactamase producing multi-drug-resistance (MDR) Klebsiella spp. had increased. Cases of MDR infections had been increased suddenly, which resulted in ineffective antimicrobials treatment. Clinicians prefer multiple combination bactericidal therapy against infection instead of single drug. Recently, an alternate approach called biofield treatment on pathogenic microorganism is reported to alter the antimicrobial susceptibility.

Biofield is a cumulative outcome of electric and magnetic field energy, exerted by the human body. However, the energy can exist in several forms such as kinetic, potential, electrical, magnetic, and nuclear. Similarly, the human nervous system consists of the energy and chemical information in the form of electrical signals. Thus, human has the ability to harness the energy from 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 useful way via biofield energy. Mr. Trivedi’s unique biofield treatment is also known as The Trivedi Effect®. In spite of countless study reports on biofield therapies [8,9], there are very few well controlled and peer-reviewed experimental studies on pathogenic or MDR microbes. According to law of mass-energy inter-conversion [10], the conversion of mass into energy is well established, but its inversion i.e. energy into mass has not yet proved scientifically. Whenever these electrical signals fluctuate with time, the magnetic field generates as per the Ampere-Maxwell law, and cumulatively known as electromagnetic field. Mr. Trivedi’s biofield treatment is well-known to change the various physicochemical characteristics of metals and ceramics [11-14]. In addition, his unique biofield treatment has considerably altered the antimicrobials susceptibility and biochemical reactions of pathogenic microbes [15-17]. In agricultural science, biofield treatment altered the growth, characteristics and yield of important medicinal plants [18-21]. On the basis of several reports on biofield treatment, present study was designed to study the impact of biofield on MDR isolate of K. oxytoca, for its antimicrobials susceptibility pattern, minimum inhibitory concentration (MIC), along with biotyping based on variation in biochemical reactions.

Materials and Methods

Experimental design and biofield treatment

MDR clinical strain of K. oxytoca was collected from stored stock cultures of clinical sample in Microbiology Lab, Hinduja Hospital, Mumbai. MDR strain was divided in two groups i.e. control and treatment. Treatment group, in sealed pack was handed over to Mr. Trivedi for biofield treatment under laboratory conditions. Mr. Trivedi provided the treatment through his energy transmission process to the treated groups without touching the samples. The biofield treated sample was returned in the similar sealed condition for further analysis on day 10 with respect to control using the standard protocols. After biofield treatment, treated sample was analyzed for antimicrobial susceptibility, biochemical reactions and biotype number using MicroScan Walk-Away® (Dade Behring Inc., USA) and Negative Break Point Combo (NBPC 30) panel with respect to control groups. The antimicrobials and biochemicals were procured from Sigma Aldrich, MA, USA.

Evaluation of antimicrobial susceptibility assay

Antimicrobial susceptibility pattern of K. oxytoca was studied using MicroScan Walk-Away® NBPC 30 as per manufacturer's instructions. The antimicrobial susceptibility pattern (S: Susceptible, I: Intermediate, and R: Resistant) and MIC were determined by observing the lowest antimicrobial concentration showing growth inhibition [22].

Biochemical reaction study

Biochemical study of K. oxytoca was determined by MicroScan Walk-Away® system in both control and treated groups [22].

Identification by biotype number

The biotype number of K. oxytoca control and treated samples were determined by MicroScan Walk-Away® processed panel data report with the help of biochemical reactions data [22].

Results and Discussion

Antimicrobial susceptibility test

Results of antimicrobial sensitivity pattern and MIC of K. oxytoca isolate are summarized in Tables 1 and 2 respectively.

Table 1: In-vitro antimicrobial susceptibility assay of multidrug resistant Klebsiella oxytoca.

The biofield treatment on MDR strain of K. oxytoca showed a significant change in sensitivity pattern of different tested antimicrobials such as amikacin and cefotetan changed from sensitive (S) to resistance (R), while aztreonam, cefotaxime, ceftazidime, and ceftriaxone sensitivity were changed from suspected extended-spectrum β-lactamases to resistance. Moreover, amoxicillin/clavulanate sensitivity changed from intermediate to resistant while piperacillin/tazobactam was changed from susceptible to intermediate as compared to control. Overall 26.67% alteration was reported out of thirty tested antimicrobials after biofield treatment. Rest of the twenty-two antimicrobials did not show any change in sensitivity after biofield treatment compared to control. MIC results showed 12.5% alteration in tested antimicrobials after biofield treatment on MDR strain of K. oxytoca. MIC value in four antimicrobials was increased out of thirty-two tested antimicrobials. Amikacin and cefotetan showed about two-folds increase in MIC value (≤ 16 to >32 µg/mL) as compared to control. Piperacillin/tazobactam showed four folds increase in MIC value (<16 to 64 µg/mL) as compared to control. Amoxicillin/k-clavulanate also showed increase MIC value with respect to control. Rest of the antimicrobials did not show any alteration in MIC values with respect to control (Table 3).

This study investigated the influence of biofield treatment on MDR strain of K. oxytoca with respect to antimicrobial sensitivity assay, and results found that biofield treatment has the potential to alter the sensitivity and MIC values of antimicrobials against biofield treated pathogen. The increased emergence of MDR strains of Klebsiella spp., in immunocompromised patients and increased infections lead to serious matter of concern worldwide. Extended spectrum β-lactamase (ESBL) producing species have still the serious problem worldwide, which may be due to continuous new drug discovery [2]. Results suggest the natural resistant pattern of MDR strain of K. oxytoca against most of the tested antimicrobials. Antimicrobial sensitivity of K. oxytoca is well supported with literature data [23]. Biofield treatment group showed significant effect on ESBL producing antimicrobials, as sensitivity after biofield treatment changed to resistant in case of aztreonam, cefotaxime, ceftazidime, and ceftriaxone. Most of the clinical strains of K. oxytoca produced chromosomal and plasmid mediated β-lactamase. Chromosomal mediated β-lactamases had the capacity to hydrolyze extended spectrum antimicrobials such as cephalosporin, and aztreonam. Mutational hyper production of β-lactamase results in a characteristic antibiogram with resistant pattern against piperacillin, cefuroxime, and aztreonam [24]. However, most of the clinical isolates of K. oxytoca have been associated with low production of β-lactamase. Biofield treatment might induce some enzymatic changes which result in significant alteration in antimicrobial sensitivity and MIC values. Resistant pattern in MDR is also associated with alteration in cell membrane, which may causes decrease uptake of antimicrobial, drug target enzyme overexpression, or alteration in drug efflux pump [25-27]. Biofield treatment in MDR K. oxytoca might alter the cell membrane permeability which results in alteration in sensitivity of tested antimicrobials.

Table 2: Minimum inhibitory concentration (MIC) tested antimicrobials of multidrug resistant Klebsiella oxytoca.

Identification of organism by biochemical reactions

Several phenotypic identification tests are available to differentiate the Klebsiella species. Experimental identification of K. oxytoca was performed using different standard biochemical reaction analysis. Adonitol, inositol, urea, and Voges-Proskauer showed negative reaction i.e. positive to negative, while cetrimide, citrate, hydrogen sulfide, and ornithine showed positive reaction i.e. negative to positive after biofield treatment as compared to control. Rest of the biochemical reactions were not altered after biofield treatment with respect to control. Overall, biochemical reactions showed the alteration of 24.24% after biofield treatment. Experimental control biochemical reaction data of K. oxytoca are well supported with literature data [2]. Biofield treatment showed a significant alteration in positive as well as negative reactions in tested biochemical, which are the basic characteristics of K. oxytoca. The standard positive biochemical reactions of K. oxytoca were reported in case of indole, lysine decarboxylase, L-sorbose, malonate, urea and Voges-Proskauer while negative reactions in ornithine decarboxylase, gas production, and citrate.

Table 3: Biochemical identification of multidrug resistant Klebsiella oxytoca.

Identification of organism by biotype number

On the basis of above biochemical changes, biotyping was performed to check the identity of microorganism after biofield treatment using an automated system. Results of biotyping found a significant changed in biotype number (7713 5272) in treated group on day 10, with respect to control (7775 4332). The organism was identified as Kluyvera ascorbata in treated group after biofield treatment as compared to control organism, K. oxytoca (Table 4). Biofield treatment on pathogenic microorganism showed significant alteration in biochemical reactions followed by altered biotype number, which are well supported with literature reports [15-17].

Biofield therapies in biomedical health care system are very popular and reported to improve human well-being with respect to several diseased conditions [28]. Increased emergence of resistant microorganisms due to widespread uses of antibiotics contributed to the spread of multidrug resistant organisms [29]. Biofield treatment is practiced by many heath care professionals as it was accepted by National Center for Complementary and Alternative Medicine (NCCAM), in complementary and alternate medicine [30]. Biofield treatment in pathogenic microorganisms had been reported to alter the antimicrobial sensitivity, phenotypic characteristics, and growth of microorganism [16,17]. It results in altered sensitivity of antimicrobials which may involve cellular changes in biofield treated K. oxytoca at molecular and/or genetic level [31]. Results showed that, biofield treatment induced changes in susceptibility pattern of antimicrobials, MIC values biochemical reactions, and biotype number of MDR strain of K. oxytoca.

Table 4: Effect of biofield treatment on multidrug resistant strain of Klebsiella oxytoca to its biotype number.

Conclusion

Altogether, the biofield treatment on MDR strain of K. oxytoca showed alteration of antimicrobial sensitivity pattern, MIC, biochemical reactions followed by biotype number. Altered biochemical reactions may be responsible for changed biotype number, and a new species was identified as Kluyvera ascorbata, as compared to control. Alteration in above standard microbiological techniques after biofield treatment might involve the changes at enzymatic or genetic level of K. oxytoca, which can be further studied at molecular level with respect to altered antimicrobial sensitivity and biotype number. Based on the study outcomes, biofield treatment could be applied to alter the sensitivity pattern of antimicrobials, against multidrug resistance strain of K. oxytoca.

Acknowledgement

The authors would like to acknowledge the whole team of PD Hinduja National Hospital and MRC, Mumbai, Microbiology Lab for their support. We are very grateful for the support of Trivedi Science™, Trivedi Master Wellness™ and Trivedi Testimonials in this research work.

Conflict of interest

The authors declare that they have no competing interest.

References

1. Gorkiewicz G (2009) Nosocomial and antibiotic-associated diarrhea caused by organisms other than Clostridium difficile Int J Antimicrob Agents 33: S37-S41

2. Podschun R, Ullmann U (1998) Klebsiella spp as nosocomial pathogens: Epidemiology, taxonomy, typing methods, and pathogenicity factors Clin Microbiol Rev 11: 589-603

3. Savino F, Cordisco L, Tarasco V, Calabrese R, Palumeri E, et al. (2009) Molecular identification of coliform bacteria from colicky breastfed infants Acta Paediatr 98: 1582-1588

4. Lin RD, Hsueh PR, Chang SC, Chen YC, Hsieh WC, et al. (1997) Bacteremia due to Klebsiella oxytoca: Clinical features of patients and antimicrobial susceptibilities of the isolates Clin Infect Dis 24: 1217-1222

5. Menard A, Harambat J, Pereyre S, Pontailler JR, Megraud F, et al. (2010) First report of septic arthritis caused by Klebsiella oxytoca J Clin Microbiol 48: 3021-3023

6. Zarate MS, Gales AC, Picao RC, Pujol GS, Lanza A, et al. (2008) Outbreak of OXY-2-producing Klebsiella oxytoca in a renal transplant unit J Clin Microbiol 46: 2099-2101

7. Sorli L, Miro E, Segura C, Navarro F, Grau S, et al. (2011) Intra- and interspecies spread of carbapenemase genes in a non-hospitalized patient Eur J Clin Microbiol Infect Dis 30: 1551-1555

8. Benor DJ (2002) Energy medicine for the internist Med Clin North Am 86: 105-125

9. Jonas WB, Crawford CC (2003) Science and spiritual healing: a critical review of spiritual healing, “energy” medicine, and intentionality Altern Ther Health Med 9: 56-61

10. Einstein A (1905) Does the inertia of a body depend upon its energy-content? Ann Phys 18: 639-641

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

12. Dhabade VV, Tallapragada RM, Trivedi MK (2009) Effect of external energy on atomic, crystalline and powder characteristics of antimony and bismuth powders Bull Mater Sci 32: 471-479

13. Trivedi MK, Patil S, Tallapragada RM (2013) Effect of biofield treatment on the physical and thermal characteristics of silicon, tin and lead powders J Material Sci Eng 2: 125

14. 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 Ind Eng Manage 4: 161

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

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

17. 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

18. Shinde V, Sances F, Patil S, Spence A (2012) Impact of biofield treatment on growth and yield of lettuce and tomato Aust J Basic Appl Sci 6: 100-105

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

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

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

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

23. Upadhyay AK, Parajuli P (2013) Extended spectrum β-lactamase producing multidrug-resistant Klebsiella species isolated at national medical college and teaching hospital, Nepal Asian J Pharm Clin Res 6: 161-164

24. Arakawa Y, Ohta M, Kido N, Mori M, Ito H, et al. (1989) Chromosomal β-lactamase of Klebsiella oxytoca, a new class A enzyme that hydrolyses broad-spectrum β-lactam antibiotics Antimicrob Agents Chemother 33: 63-70

25. Tenover FC (2006) Mechanisms of antimicrobial resistance in bacteria Am J Infect Control 119: S3-S10

26. He X, Li S, Kaminskyj SG (2013) Using Aspergillus nidulans to identify antifungal drug resistance mutations Eukaryot Cell 13: 288-294

27. Alekshun MN, Levy SB (2007) Molecular mechanisms of antibacterial multidrug resistance Cell 128: 1037-1050

28. Turner JG, Clark AJ, Gauthier DK, Williams M (1998) The effect of therapeutic touch on pain and anxiety in burn patients J Adv Nurs 28: 10-20

29. Okeke IN, Laxminarayan R, Bhutta ZA, Duse AG, Jenkins P, et al. (2005) Antimicrobial resistance in developing countries Part I: Recent trends and current status Lancet Infect Dis 5: 481-493

30. Koithan M (2009) Introducing complementary and alternative therapies J Nurse Pract 5: 18-20

31. 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.