Raman spectroscopy reveals distinct differences between two closely related bacterial strains, Mycobacterium indicus pranii and Mycobacterium intracellulare

Verma, Taru; Podder, Santosh; Mehta, Mayank R.; Singh, Sarman; Singh, Amit; Umapathy, Siva; Nandi, Dipankar

doi:10.1007/s00216-019-02197-z

Cited by 14 publications

(7 citation statements)

References 45 publications

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“…It has also been used to study the biology of mycobacteria ( Buijtels et al, 2008 , Kumar et al, 2020 , Perumal et al, 2018 , Stöckel et al, 2017 , Stöckel et al, 2015 , Verma et al, 2019 ). Raman spectroscopy also has potential as a diagnostic tool, as it allows the identification of mycobacteria to the species level ( Buijtels et al, 2008 , Stöckel et al, 2017 , Stöckel et al, 2015 , Verma et al, 2019 ), and even to determine the viability of the identified bacilli ( Kumar et al, 2020 ). We generated the Raman spectra from M. chelonae planktonic, as well as 5 day (Biofilm t1) and 10 day old biofilms (Biofilm t2) ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Biofilms of the non-tuberculous Mycobacterium chelonae form an extracellular matrix and display distinct expression patterns

et al. 2020

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Mycobacterium chelonae is an environmental, non-tuberculous mycobacterial species, capable of causing infections in humans. Biofilm formation is a key strategy used by M. chelonae in colonising niches in the environment and in the host. We studied a water-air interface (pellicle) biofilm of M. chelonae using a wide array of approaches to outline the molecular structure and composition of the biofilm. Scanning electron micrographs showed that M. chelonae biofilms produced an extracellular matrix. Using a combination of biochemical analysis, Raman spectroscopy, and fluorescence microscopy, we showed the matrix to consist of proteins, carbohydrates, lipids and eDNA. Glucose was the predominant sugar present in the biofilm matrix, and its relative abundance decreased in late (established) biofilms. RNA-seq analysis of the biofilms showed upregulation of genes involved in redox metabolism. Additionally, genes involved in mycolic acid, other lipid and glyoxylate metabolism were also upregulated in the early biofilms.

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Section: Resultsmentioning

confidence: 99%

Biofilms of the non-tuberculous Mycobacterium chelonae form an extracellular matrix and display distinct expression patterns

et al. 2020

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“…Samples were processed for AFM imaging as described previously [8,25,26]. Briefly, bacterial cultures were grown for 6 hours at 37 C, under different conditions, after which they were centrifuged at 5000 rpm for 5 minutes.…”

Section: Atomic Force Microscopy (Afm)mentioning

confidence: 99%

“…After washing with sterile water, 10 μl of the bacterial pellet was drop-cast onto a clean and dry magnesium fluoride coverslip and air-dried for 5 to 10 minutes. Raman spectra were acquired from the dried bacterial drop using the Renishaw InVia Raman Microscope (Renishaw, UK) equipped with 785 nm laser [26][27][28]. The laser was focussed onto the sample using a 50× objective with numerical aperture of 0.75 (Leica, Germany).…”

Section: Sample Preparation and Raman Spectral Acquisitionsmentioning

confidence: 99%

Profiling antibiotic resistance in Escherichia coli strains displaying differential antibiotic susceptibilities using Raman spectroscopy

Verma

Annappa

Singh

et al. 2020

Journal of Biophotonics

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The rapid identification of antibiotic resistant bacteria is important for public health. In the environment, bacteria are exposed to subinhibitory antibiotic concentrations which has implications in the generation of multi-drug resistant strains. To better understand these issues, Raman spectroscopy was employed coupled with partial least squares-discriminant analysis to profile Escherichia coli strains treated with sub-inhibitory concentrations of antibiotics. Clear differences were observed between cells treated with bacteriostatic (tetracycline and rifampicin) and bactericidal (ampicillin, ciprofloxacin, and ceftriaxone) antibiotics for 6 hr: First, atomic force microscopy revealed that bactericidal antibiotics cause extensive cell elongation whereas short filaments are observed with bacteriostatic antibiotics. Second, Raman spectral analysis revealed that bactericidal antibiotics lower nucleic acid to protein (I 812 /I 830) and nucleic acid to lipid ratios (I 1483 / I 1452) whereas the opposite is seen with bacteriostatic antibiotics. Third, the protein to lipid ratio (I 2936 /I 2885 and I 2936 /I 2850) is a Raman stress signature common to both the classes. These signatures were validated using two mutants, Δlon and ΔacrB, that exhibit relatively high and low resistance towards antibiotics, respectively. In addition, these spectral markers correlated with the emergence of phenotypic antibiotic resistance. Overall, this study demonstrates the efficacy of Raman spectroscopy to identify resistance in bacteria to sub-lethal concentrations of antibiotics.

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“…Raman spectra were collected using a Renishaw InVia Raman spectrometer equipped with an upright microscope. The spectra were recorded from the air-dried drop using two different lasers: a 785 nm diode laser (power ∼35 mW) and a 633 nm He–Ne laser (power ∼8 mW), and the spectrometer was equipped with 1200 grooves/mm grating. ,− The spectrometer calibration was done using a silicon peak at 520.5 cm –1 . Raman spectral collection was done using 100× objective (N.A.…”

Section: Methodsmentioning

confidence: 99%

Herbicides 2,4-Dichlorophenoxy Acetic Acid and Glyphosate Induce Distinct Biochemical Changes in E. coli during Phenotypic Antibiotic Resistance: A Raman Spectroscopic Study

et al. 2022

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Antibiotic resistance is a major global health concern. The increased use of herbicides may lead to multiple antibiotic resistance in bacteria. Conventional techniques for diagnosing antibiotic resistance are laborious, time-intensive, expensive, and lack information about antibiotic susceptibility. On the other hand, Raman spectroscopy is a rapid, label-free, noninvasive alternative to traditional techniques to detect antibiotic resistance. In this study, two popular herbicides 2,4-dichlorophenoxy acetic acid (2,4-D) and N-(phosphonomethyl)glycine (glyphosate) were used to study their effects on the emergence of antibiotic resistance. The Escherichia coli wild-type (WT) MG1655 strain and two isogenic mutants, Δlon and ΔacrB, were used together with Raman spectroscopy. The WT E. coli is sensitive to antibiotics, but exposure to both herbicides induces antibiotic resistance. Using an excitation wavelength of 785 nm, the intensity ratios (e.g., I 740/I 785, I 740/I 1003, I 1480/I 1445, I 2934/I 2868, and I 2934/I 2845) were identified as biomarkers to study the induction of antibiotic resistance in bacteria but not NaCl-mediated stress. Using an excitation wavelength of 633 nm, the peak intensity at 740 cm–1 assigned to cytochrome bd decreases under antibiotic stress but increases upon exposure to both herbicides and antibiotics, indicating the development of resistance. Thus, this study can be applied to monitor antibiotic resistance using Raman spectroscopy.

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Raman spectroscopy reveals distinct differences between two closely related bacterial strains, Mycobacterium indicus pranii and Mycobacterium intracellulare

Cited by 14 publications

References 45 publications

Biofilms of the non-tuberculous Mycobacterium chelonae form an extracellular matrix and display distinct expression patterns

Biofilms of the non-tuberculous Mycobacterium chelonae form an extracellular matrix and display distinct expression patterns

Profiling antibiotic resistance in Escherichia coli strains displaying differential antibiotic susceptibilities using Raman spectroscopy

Herbicides 2,4-Dichlorophenoxy Acetic Acid and Glyphosate Induce Distinct Biochemical Changes in E. coli during Phenotypic Antibiotic Resistance: A Raman Spectroscopic Study

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