Type‐IV pili spectroscopic markers: Applications in the quantification of piliation levels inMoraxella boviscells by a FT‐IR ANN‐based model

Abstract

Type‐IV pili are cell surface organelles found in a wide variety of Gram‐negative bacteria. They have traditionally been detected by electron microscopy and ELISA techniques. However, these methodologies are not appropriate for the rapid discrimination and quantification of piliated and nonpiliated cells in industrial or field conditions. Here, the analysis of FT‐IR spectra of piliated, nonpiliated and sheared Moraxella bovis cells, together with purified pili suspensions spectra, allowed the identification of 3 IR regions associated to spectroscopic markers of Type‐IV pili: 1750–1600, 1450–1350 and 1280–950 cm–1. Such IR‐specific markers were found for piliated cells grown in different culture systems (liquid or solid media), independently of the strain or pili serotype. They were also sensitive to pili expression levels. Therefore, on the bases of these specific spectral features, an FT‐IR ANN‐based model was developed to classify piliation levels in 5 distinct groups. An overall classification rate of almost 90% demonstrates the strong potential of the ANN system developed to monitor M. bovis cultures in vaccine production. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Analytical techniques for deoxynivalenol detection and quantification in wheat destined for the manufacture of commercial products

“…C-O-C, C-OH, C-H; cf. Bosch et al, 2010;Helm and Naumann, 1995;Kacurakova et al, 2000;Naumann, 2000).…”

IR and Raman Spectroscopy for Pathogen Detection

“…Especially, the inspiring works of Naumann and coworkers reintroduced FT-IR methods in combination with chemometric tools for in situ analyses of bacterial cells, which resulted in a boom in scientific publications over the past 20 years [8,9,15]. Since then, FT-IR spectroscopy has been successfully used to study, among others, biofilm formation, cellular components in bacteria, microbial response to stress, injury, and inactivation, as well as the development of antibiotic resistance [16][17][18]. Emphasis was also placed on the detection, discrimination, classification, and identification of bacteria -especially foodborne pathogens -such as Bacillus [19,20], Brucella [21], cyanobacteria [22], E. coli [23,24], lactobacilli [25], Listeria [26,27], Mycobacteria [28], Streptococcus [29], Salmonella [30,31], Staphylococcus [32,33], and Yersinia [34,35].…”

Evaluation of biofilm-forming capacity of Moraxella bovis, the primary causative agent of infectious bovine keratoconjunctivitis

“…Stock cultures were prepared with cells grown on TSAB for 24 h at 37 8C, and kept at À80 8C in 20% (v/v) glycerol. After thawing, the presence or absence of pili in each stock culture was confirmed by electron microscopy, enzyme-linked immunosorbent assay, and Fourier-transform infrared (FT-IR) spectroscopy (Bosch et al, 2010;Prieto et al, 2003Prieto et al, , 2008. Furthermore, at the end of biofilms cultures, the piliation level of sessile cells was controlled by ELISA technique and/ or FT-IR spectroscopy.…”