Chicken, beams, and Campylobacter: rapid differentiation of foodborne bacteria via vibrational spectroscopy and MALDI-mass spectrometry

Abstract

Campylobacter species are one of the main causes of food poisoning worldwide. Despite the availability of established culturing and molecular techniques, due to the fastidious nature of these microorganisms, simultaneous detection and species differentiation still remains challenging. This study focused on the differentiation of eleven Campylobacter strains from six species, using Fourier transform infrared (FT-IR) and Raman spectroscopies, together with matrix-assisted laser desorption ionisation-time of flight-mass spectrometry (MALDI-TOF-MS), as physicochemical approaches for generating biochemical fingerprints. Cluster analysis of data from each of the three analytical approaches provided clear differentiation of each Campylobacter species, which was generally in agreement with a phylogenetic tree based on 16S rRNA gene sequences. Notably, although C. fetus subspecies fetus and venerealis are phylogenetically very closely related, using FT-IR and MALDI-TOF-MS data these subspecies were readily differentiated based on differences in the lipid (2920 and 2851 cm(-1)) and fingerprint regions (1500-500 cm(-1)) of the FT-IR spectra, and the 500-2000 m/z region of the MALDI-TOF-MS data. A finding that was further investigated with targeted lipidomics using liquid chromatography-mass spectrometry (LC-MS). Our results demonstrate that such metabolomics approaches combined with molecular biology techniques may provide critical information and knowledge related to the risk factors, virulence, and understanding of the distribution and transmission routes associated with different strains of foodborne Campylobacter spp.

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

“…Species FTIRS HCA [51] Klebsiella spp. Species FTIRS HCA, ANN [51,92] Proteus mirabilis Species FTIRS HCA, ANN [51,92] Lipopolyssacharide structure FTIRS-ATR HCA, PCA, RF [85] Geobacillus caldoxylosilyticus Species FTIRS HCA [53] Thermobrachium celere Species FTIRS HCA [53] Thermoanaerobacter ethanolicus Species FTIRS HCA [53] Micrococcus luteus Species FTIRS PCA, HCA [53,54] Pantoea agglomerans Species FTIRS PCA, HCA [54] Legionella pneumophila Serogroup FTIRS - [66] Brucella spp. Biovars FTIRS PCA, LDA [78] Xanthomonas oryzae Pathovars FTIRS - [79] Campilobacter spp.…”

“…Species FTIRS, FTIRS-ATR HCA, PCA, DFA, ANN [43][44][45] Acinetobacter baumannii Sequence type FTIRS-ATR PLSDA [69,70] Burkholderia cepacia complex Species FTIRS PLS, PCA, SIMCA, ANN [45,46] Ribopatterns FTIRS PCA, PLS-DA [46] Pseudomonas spp. Species FTIRS HCA, ANN, PCA, SIMCA [45,49,53,86,92] Stenotrophomonas maltophilia Species FTIRS PCA, ANN [45,55] Ralstonia pickettii Species FTIRS ANN [45] Achromobacter spp Species FTIRS ANN [45] Yersinia spp. Species FTIRS, FTIRS-ATR ANN, SIMCA, PCA, CVA [47,50,52] Yersinia enterocolitica Serotype FTIRS ANN [47] Biotype FTIRS ANN [47] Yersinia ruckeri Biotype FTIRS ANN [80] Oscillatoria limosa Species FTIRS-ATR PCA, ANN [48] Arthrospira platensis Species FTIRS-ATR PCA, ANN [48] Phormidium spp.…”

“…Species FTIRS HCA, ANN [31,32,39,40,53] Enterococcus spp. Species FTIRS, Diffuse Reflectance FTIRS HCA, ANN [33,36,[39][40][41]51,92] Listeria monocytogenes Serotype FTIRS CVA, ANN, HCA [21,25,56,57] Epidemic clones FTIRS CVA, LDA [76] Halotype FTIRS HCA, CVA [57] Intact/Injured FTIRS CVA, LDA, PCA [76,84] Enterococcus faecium PFGE type FTIRS PLSDA [71] Streptococcus spp. Species FTIRS, Diffuse Reflectance FTIRS ANN, HCA [4,34,36,39,40] Streptococcus pneumonia Serotype FTIRS [64] Staphylococcus spp.…”

“…Sulphate and thiosulphate reducing bacteria Genus FTIRS, FTIRS-ATR HCA, PCA [16,17] Alicylobacillus Genus FTIRS PCA, SIMCA [20] Strain FTIRS-HATR LDA, PCA [75] Photorhabdus Genus FTIRS-ATR HCA, PCA [18] Xenorhabdus Genus FTIRS-ATR HCA, PCA [18] Arcanobacterium Genus FTIRS ANN [19] Actinomyces Genus FTIRS ANN [19] Trueperella Genus FTIRS ANN [19] Species FTIRS ANN [19] Bacillus Genus FTIRS PCA, SIMCA [20] Species FTIRS, FTIRS-ATR, Diffuse Reflectance-FTIRS PCA, HCA, DFA, CVA, SIMCA [26][27][28][29][30]49,50,54,55] Vegetative/Sporulated FTIRS PCA, CART [83] Escherichia Genus FTIRS-ATR HCA, PCA [18] Species FTIRS, FTIRS-ATR HCA, ANN, PCA, SIMCA, CVA [49][50][51][52][53][54]86,92] Escherichia coli Sequence type FTIRS, FTIRS-ATR PC-DFA, PLSDA, HCA, SIMCA [67,68] Strain FTIRS PCA, SIMCA, CVA [49,50,52,73] MLVA profile FTIRS HCA, CVA [72] Listeria spp. Species FTIRS, FTIRS-ATR, FTIR microspectroscopy CVA, ANN, SCDA, PLSRDA, PCA [5,[21][22][23][24][25]…”

“…In contrast, similar results were obtained with FTIRS and MALDI-TOF MS regarding the discrimination of Klebsiella oxytoca [87] and Campylobacter strains. [92] FTIRS was compared with Raman, Raman micro-spectroscopy, atomic force microscopy (AFM), and IF. Raman spectroscopy exhibits some known advantages over IR spectroscopy such as the lower interference of water and narrower bands.…”