Using Laser Induced Breakdown Spectroscopy (LIBS) for Investigation Different Growth Phases of Escherichia coli BL21

Document Type : Original Article

Authors

1 Science and Technology Center of Laser and Optics, Faculty of Basics Science, Imam Hossein University, Tehran, Iran

2 Associate Professor of Imam Hossein Comprehensive University, Tehran, Iran.

3 Assistant Professor, Malik Ashtar University of Technology, Tehran, Iran.

4 Assistant Professor، Imam Hossein Comprehensive ، University, Tehran, Iran.

Abstract

Due to the limitations of conventional biological identification methods, spectroscopic methods, including laser induced breakdown spectroscopy, have been used to characterize the genus, species and strains of different bacteria in the static phase of growth. Due to the increading importance of biological problems, the characterization, detection and classification of bacterial specimens has been considered by researchers in the military, industrial and medical fields. In this paper, in order to monitor the changes in the elemental structure of bacteria in the growth process, the constituent elements of the two phases of exponential and static growth of Escherichia coli were studied and compared based on this spectroscopic method. In this experiment, Nd: YAG laser with 100 mJ pulse energy, spectrometer and CCD detector were used. The industrial strain of gram-negative Escherichia coli BL21 bacteria was selected. Bacteria were selected at three different times (2.5, 4 and 6 hours after inoculation) during the growth process in broth, at optical density of 0.4 and 0.8 (exponential growth phase) and optical density of 1.2 (Static growth phase), respectively, and after dry freezing, under pressure has been pressed into tablets. Considering the chemical structure of Escherichia coli, the elements of phosphorus, carbon, calcium, sodium, magnesium, potassium, oxygen, nitrogen and hydrogen were considered in the sample. The results showed that Escherichia coli bacteria in the growth process have different patterns of recognizable elemental composition in its structure in the growth process. It is predicted that due to the change in the composition of structural elements of bacteria in growth phases, the use of this spectroscopic method to identify bacteria will be dependent on the growth phase. The results of this study showed that this method can play an effective role in the development of polyphasic methods to indentify bacteria, determine the composition of the culture medium as well as optimizing and evaluating the production conditions in this industrial strain.

Keywords


  1. Gottfried, J. L.; Delucla F. C.; Chase, A.; Munson, A.; Miziolek, W. “Standoff Detection of Chemical and Biological Threats Using Laser-Induced Breakdown Spectroscopy”; Appl. Spectrosc. 2008, 62, 353-363.##
  2. Dudrange, L.; Adam, L. D.; Amouroux, J. “Time-Resolved Laser-Induced Breakdown Spectroscopy: Application for Fluorine, Chlorine, Sulfur, and Carbon in Air”; Appl. Spectrosc. 1998, 52, 1321–1327.##
  3. Morel, S.; Leone N.; Adam P.; Amouroux, J. “Detection of Bacteria by Time-Resolved Laser-Induced Breakdown Spectroscopy”; Appl. Opt. 2003, 42, 6184-6191.##
  4. Mohaidat, Q. I. “Laser-Induced Breakdown Spectroscopy (LIBS): An Innovative Tool for Studying Bacteria” PhD. Diss. Detroit, Michigan, Wayne State Univ, 2011.##
  5. Farooq, W. A.; Atif, M.; Tawfik, W.; Alsalhi, M. S.; Alahmed, Z. A.; Sarfraz M. “Study of Bacterial Samples Using Laser-Induced Breakdown Spectroscopy”; Plasma Sci. Technol. 2014, 16, 1141-1146.##
  6. Kim, T.; Specht, Z. G.; Vary, P. S.; Lin, C. T. “Spectral Fingerprints of Bacterial Strains by Laser-Induced Breakdown Spectroscopy”; J. Phys. Chem. B 2004, 17, 5477-5482.##
  7. Rehse, S. J.; Diedrich, J.; Palchaudhuri, S. “Identification and Discrimination of Pseudomonas Aeruginosa Bacteria Grown in Blood and Bile by Laser-Induced Breakdown Spectroscopy”; Spectrochim. Acta Part B 2007, 62, 1169-1176.##
  8. Mohaidat, Q. I.; Sheikh, K.; Palchaudhuri, S. ; Rehse, S. J. “Pathogen Identification with Laser-Induced Breakdown Spectroscopy: The Effect of Bacterial and Biofluid Specimen Contamination”; Appl. Opt. 2012, 51, 99-107.##
  9. Diedrich, J.; Rehse, S. J. ; Palchaudhuri S. “Escherichia Coli Identification and Strain Discrimination Using Nanosecond Laser-Induced Breakdown Spectroscopy”; Appl. Phys. Lett. 2007, 90, 163901-3.##
  10. Sivakumar, V.; Sujatha, N. U.; Nilesh, J. V.; Srikanth, P.; Arumugam, I. “Bacterial Strain Discrimination Using a Low-Cost Laser-Induced Breakdown Spectroscopy Technique under Optimized Growth Conditions”; Proceedings SPIE Photonics Euroope, Clinical Biophotonics 2020, 11362.##
  11. Rehse, S. J.; Mohaidat, Q. I.; Palchaudhuri, S. “Towards the Clinical Application of Laser-Induced Breakdown Spectroscopy for Rapid Pathogen Diagnosis: The Effect of Mixed Cultures and Sample Dilution on Bacterial Identification”; Appl. Phys. 2010, 49, 27-35.##
  12. Dixon, P. B.; Hahn, D. W. “Feasibility of Detection and Identification of Individual Bioaerosols Using Laser-Induced Breakdown Spectroscopy”; Analitical Chemistry 2005, 77, 631-638.##
  13. Multari, R.; Cremers, D. A.; Dupre, J. M. Gustafson, J. E. “The Use of Laser-Induced Breakdown Spectroscopy for Distinguishing Between Bacterial Pathogen Species and Strains”; Appl. Spectrosc. 2010, 64, 750-759.##
  14. Malenfant, D. J.; Gillies, D. J.; Rehse, S. J. “Bacterial Suspensions Deposited on Microbiological Filter Material for Rapid Laser-Induced Breakdown Spectroscopy Identification”; Appl. Spectrosc. 2016, 70, 485-493.##
  15. Gottfried, J. L.; De Lucia, Jr. F. C.; Munson, C. A.; Miziolek A. W. “Double-Pulse Standoff Laser-Induced Breakdown Spectroscopy for Versatile Hazardous Materials Detection”; Spectrochim Acta Part B 2007, 62, 1405-1411.##
  16. Mohaidat, Q.; Palchaudhuri, S.; Rehse, S. J. “The Effect of Bacterial Environmental and Metabolic Stresses on a LIBS-Based Identification of Escherichia coli and Streptococcus Viridans”; Appl. Spectrosc. 2011, 65, 386-392.##
  17. Sing, V. K.; Sharma, J.; Phathak, A. K,; Ghany, C. T.; Gondal, M. A. “Laser Induced Breakdown Spectroscopy (LIBS): A Novel Technology for Identifying Microbes Causing Infectious Diseases”; Biophysical Reviews 2018, 10, 1221-1239.##
  18. Shakouri moghaddam, A.; Shorsheyni, S. Z.; Shirvani mahdavi, H. R.; Khodabakhshi, Z. “Identification and Discrimination Bacterias of Gram Positive and Negative using Laser-Induced Break Down Spectroscopy (LIBS)”; 24th ICOP, 2018, 901-904 (In Persian).##
  19. Rehse, S. J. “A Review of the Use of Laser-Induced Breakdown Spectroscopy for Bacterial Classification, Quantification, and Identification”; Spectrochim Acta Part B. 2019, 154, 50-69.
  20. NIST.gov.##
  21. Paulick, A. E. “Development of Laser Induced Breakdown Spectroscopy as Rapid Diagnostic Tool for Bacterial Infection” PhD. Diss., Windsor Univ 2018.##
Volume 12, Issue 4 - Serial Number 46
February 2022
Pages 409-418
  • Receive Date: 01 November 2021
  • Revise Date: 14 December 2021
  • Accept Date: 22 December 2021
  • Publish Date: 20 February 2022