Fluorescence Spectroscopy as a Promising Tool for a Polyphasic Approach to Pseudomonad Taxonomy

Abstract

Fluorescence spectroscopy is an emerging tool for the analysis of biomolecules from complex matrices. We explored the potentialities of the method for the pseudomonad taxonomic purpose at the genus and species level. Emission spectra of three intrinsic fluorophores (namely, NADH, tryptophan, and the complex of aromatic amino acids and nucleic acid) were collected from whole bacterial cells. Their comparisons were performed through principal component analysis and factorial discriminant analysis. Reference strains from the Xanthomonas, Stenotrophomonas, Burkholderia, and Pseudomonas genera were well separated, with sensitivity and selectivity higher than 90%. At the species level, P. lundensis, P. taetrolens, P. fragi, P. chlororaphis, and P. stutzeri were also well separated, in a distant group, from P. putida, P. pseudoalcaligenes, and P. fluorescens. These results are in agreement with the generally admitted rRNA and DNA bacterial homology grouping but they also provide additional information about strain relatedness. In the case of environmental isolates, the method allows good discrimination, even for strains for which ambiguity still remained after PCR and API 20NE identification. Rapid, easy to perform, and low cost, fluorescence spectroscopy provides substantial information on cell components. Statistical analysis of collected data allows in-depth comparison of strains. Our results strongly support the view that fluorescence spectroscopy fingerprinting can be used as a powerful tool in a polyphasic approach to pseudomonad taxonomy.

Fluorescence Spectroscopy Measurement for Quality Assessment of Food Systems—a Review

“…These findings were reinforced by the high correlation (using CCA) observed between the data sets obtained from the metabolic profiling and fluorescence spectroscopy. The obtained results were fully supported by the same research group (Tourkya et al 2009) since good discrimination, even for strains for which ambiguity still remained after PCR and Analytical Profile Index 20 NE identification. Saito (2009) depicted that Napa cabbage (Brassica rapa L.) laser-induced fluorescence (LIF) spectra of a normal core and a rotten core show greater intensities in the green wavelength range of the LIF spectrum of the rotten core in comparison to that of the normal core.…”

“…The emission spectrum Boubellouta and Dufour (2008), Dufour and Riaublanc (1997), Dufour et al (2000), Hammami et al (2010), Herbert et al (1999Herbert et al ( , 2000, Dufour (2003, 2006), Karoui et al 2004a, b, 2005a, b, c, 2006b, e, 2007c, Kulmyrzaev et al (2005 Baunsgaard et al (2000a, b), Bro (1999), Karoui et al (2007a, Munck et al (1998), Nørgaard (1995, Ruoff et al (2006) Noh and Lu (2007), Seiden et al (1996) Ammor et al (2004, Dufour (2002, 2004), Leriche et al (2004), Tourkya et al (2009 Collagen Egelandsdal et al (1996Egelandsdal et al (...…”

Synchronous front-face fluorescence spectroscopy as a promising tool for the rapid determination of spoilage bacteria on chicken breast fillet

“…Considering spectroscopic techniques, fluorescence spectroscopy have also recently been used for identification of bacteria (Ammor, Yaakoubi, Chevallier, & Dufour, 2004;Leblanc & Dufour, 2002), separation of bacteria from yeast (Bhatta, Goldys, & Learmonth, 2006) and differentiation among microorganisms belonging to different taxonomical families (Giana, Silveira, Zangaro, & Pacheco, 2003;Leblanc & Dufour, 2002;Sorrell, Tribble, Reinisch, Werkhaven, & Ossoff, 1994;Tourkya, Boubellouta, Dufour, & Leriche, 2009). A lot of endogenous molecules such as proteins, vitamins, co-enzymes of bacteria are fluorescent thus making fluorescence spectroscopy a valuable technique for the determination of microbial spoilage in meat.…”

An Overview of the Evolution of Infrared Spectroscopy Applied to Bacterial Typing

“…Regarding IF spectroscopy, several studies have been published claiming the high potential of the technique in the context of bacterial typing. [94][95][96][97] However, a recent study [98] directly compared the performance of IF spectroscopy with FTIRS and MALDI-TOF MS for Acinetobacter species identification. The study concluded that IF spectroscopy has a significant lower potential for bacterial typing.…”