Achim Kohler

Extended Multiplicative Signal Correction as a Tool for Separation and Characterization of Physical and Chemical Information in Fourier Transform Infrared Microscopy Images of Cryo-sections of Beef Loin

Extended multiplicative signal correction (EMSC) is used to separate and to characterize physical and chemical information in spectra from Fourier transform infrared (FT-IR) microscopy. This appears especially useful for applications in infrared spectroscopy where the scatter variance in spectra changes with the chemical variance in the sample set. In these cases the chemical information of specific bands that are assigned to functional groups is easier to interpret when the scatter information is removed from the spectra. We show that scatter (physical) information in FT-IR spectra of heat-treated beef loin is related to chemical changes due to heat treatment. This information is caused by textural changes induced by the heat treatment and expressed by physical effects as the optical path length. The chemical absorbance changes introduced in the FT-IR spectra due to heat treatment are shifts in the protein region of the infrared spectrum caused by changes in the secondary structure of the proteins. If the scatter and the chemical information is not separated properly, scatter information may erroneously be interpreted as chemical information.

Explorative Multifactor Approach for Investigating Global Survival Mechanisms of Campylobacter jejuni under Environmental Conditions

ABSTRACT Explorative approaches such as DNA microarray experiments are becoming increasingly important in microbial research. Despite these major technical advancements, approaches to study multifactor experiments are still lacking. We have addressed this problem by using rotation testing and a novel multivariate analysis of variance (MANOVA) approach (50-50 MANOVA) to investigate interacting experimental factors in a complex experimental design. Furthermore, a new rotation testing based method was introduced to calculate false-discovery rates for each response. This novel analytical concept was used to investigate global survival mechanisms in the environment of the major food-borne pathogen C. jejuni. We simulated nongrowth environmental conditions by investigating combinations of the factors temperature (5 and 25°C) and oxygen tension (anaerobic, microaerobic, and aerobic). Data were generated with DNA microarrays for information about gene expression patterns and Fourier transform infrared (FT-IR) spectroscopy to study global macromolecular changes in the cell. Microarray analyses showed that most genes were either unchanged or down regulated compared to the reference (day 0) for the conditions tested and that the 25°C anaerobic condition gave the most distinct expression pattern with the fewest genes expressed. The few up-regulated genes were generally stress related and/or related to the cell envelope. We found, using FT-IR spectroscopy, that the amount of polysaccharides and oligosaccharides increased under the nongrowth survival conditions. Potential mechanisms for survival could be to down regulate most functions to save energy and to produce polysaccharides and oligosaccharides for protection against harsh environments. Basic knowledge about the survival mechanisms is of fundamental importance in preventing transmission of this bacterium through the food chain.

Determination of C22:5 and C22:6 marine fatty acids in pork fat with Fourier transform mid-infrared spectroscopy

Fatty acids in samples (n=74) of pork adipose tissue were measured with a Fourier transform mid-infrared (FT-MIR) spectrometer and by gas chromatography. The measured absorption spectra provided information to estimate partial least squares regression models for fatty acid groups, the iodine value and several fatty acids. The iodine values were predicted with correlation coefficient R=0.996 and root mean square error of cross-validation RMSECV=0.658. The sum of the two marine fatty acids of main interest, C22:5n3+C22:6n3, were predicted with R=0.982 and RMSECV=0.062. The K nearest neighbours procedure successfully classified the samples in three classes, depending on their proportions of marine fatty acids. Application of fat and absorption measurements were rapid, requiring less than 5 min of labour per sample. The results reported in this paper demonstrate that FT-MIR measurements can serve as a rapid method to determine marine fatty acids in pork fat.

Characterizing mixed microbial population dynamics using time-series analysis.

Due to a general shortage of temporal population data, dynamic structures in microbial communities remain largely unexplored. Knowledge of community dynamics is, however, essential for understanding the mechanisms by which microbes interact. Here, we have used a computational approach for quantification of bacteria in multispecies populations, generating data for time-series modeling. Moreover, we have used online FR-IR spectroscopy to monitor the main metabolic processes. The approach enabled us to provide a functional description of the parameters governing the population dynamics in a three-species model bacterial community, demonstrating density-dependent regulation, interspecies competition and even a case of cooperation between two species. Since the field of microbial ecology has yet to embrace many of the concepts and methods developed for the study of ecology of higher plants and animals, the realization that microbial systems can be analyzed within the same conceptual framework as other ecosystems is of fundamental importance.