Identification of Trueperella pyogenes Isolated from Bovine Mastitis by Fourier Transform Infrared Spectroscopy

Abstract

The present study was designed to investigate the potential of Fourier transform infrared (FT-IR) spectroscopy to identify Trueperella (T.) pyogenes isolated from bovine clinical mastitis. FT-IR spectroscopy was applied to 57 isolates obtained from 55 cows in a period from 2009 to 2012. Prior to FT-IR spectroscopy these isolates were identified by phenotypic and genotypic properties, also including the determination of seven potential virulence factor encoding genes. The FT-IR analysis revealed a reliable identification of all 57 isolates as T. pyogenes and a clear separation of this species from the other species of genus Trueperella and from species of genus Arcanobacterium and Actinomyces. The results showed that all 57 isolates were assigned to the correct species indicating that FT-IR spectroscopy could also be efficiently used for identification of this bacterial pathogen.

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

“…Principal component analysis (PCA) as well as a HCA revealed three distinct clusters, each containing isolates of one single genus. In 2014, Nagib et al [19] discriminated bovine mastitis Trueperella pyogenes isolates belonging to Arcanobacterium and Actinomyces genus with an artificial neural network (ANN). More recently, Al-Holy and co-workers [20] discriminated Bacillus from Alicylobacillus isolates through PCA and soft independent modeling of class analogy (SIMCA).…”

“…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]…”

“…Other works proved the ability of FTIRS combined with chemometrics for Carnobaterium typing, [37] to discriminate species of Actinomycetes, [38] lactic acid bacteria, [39,40] food-borne pathogens commonly found in cheese and milk, [41,42] and pathogens of domestic ruminants and pigs causing mastitis such as Trueperella spp. [19] Regarding Gram-negative bacteria, only Acinetobacter, Burkholderia, and Yersinia genus were explored for species discrimination exclusively within the genus itself. Acinetobacter genus comprises several genomic and closely related species and their accurate identification remains a challenge for microbiologists.…”

Chemometric analysis of attenuated total reflectance infrared spectra of Proteus mirabilis strains with defined structures of LPS

“…6 FT-IR analyzes the chemical bonding in the total biochemical composition of the cell, including the cell wall, membrane (composed of phospholipid bilayer, peptidoglycan and LPS) and the cytoplasm (fatty acids, water, nucleic acids, proteins and polysaccharides). [4][5][6][7][8] The most commonly used method is the attenuated total reflectance FT-IR technique (ATR FT-IR). When using this, it is not required in samples to suspend and then dehydrate colonies, which results in simpler (direct) sample preparation, is easy to implement, allows the analysis of small quantities of biomass, and requires no consumables or reagents, and reduces costs.…”

“…7,8,13,33 Funding The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by a grant from the National Science Center, Poland issued on the basis of decision number DEC-2012/07/N/NZ6/04118. Some of the experiments were run on apparatus purchased with European Union grant 307 under 2.2 Innovation Industry.…”