Decision Support for the Comparative Evaluation and Selection of Analytical Methods: Detection of Genetically Modified Organisms as an Example

The selection of the best-fit-for-purpose analytical method to be implemented in the laboratory is difficult due to availability of multiple methods, targets, aims of detection, and different kinds and sources of more or less reliable information. Several factors, such as method performance, practicability, cost of setup, and running costs need to be considered together with personnel training when selecting the most appropriate method. The aim of our work was to prepare a flexible multicriteria decision analysis model suitable for evaluation and comparison of analytical methods used for the purpose of detecting and/or quantifying genetically modified organisms, and to use this model to evaluate a variety of changing analytical methods. Our study included selection of PCR-, isothermal-, protein-, microarray-, and next-generation sequencing-based methods in simplex and/or multiplex formats. We show that the overall result of their fitness for purpose is relatively similar; however, individual criteria or a group of related criteria exposed more substantial differences between the methods. The proposed model of this decision support system enables easy modifications and is thus suitable for any other application of complex analytical methods.     

Implementing lean practices in manufacturing SMEs: testing ‘critical success factors’ using Necessary Condition Analysis

Lean practices are known to increase operational performance. Previous research has identified critical success factors for implementing lean practices. This research aims to examine the extent to which success factors are critical for various degrees of lean practice implementation. Using multiple-respondent self-assessments from 33 Dutch manufacturing small and medium-sized enterprises (SMEs), we conducted a Necessary Condition Analysis. Our findings indicated that the criticality of success factors is progression dependent. In the initial stages of the lean journey, SMEs could improve their lean practices in a bottom-up manner through local factors such as a learning focus, improvement training and support congruence. When lean practices are more advanced, some company-wide factors must be present: top management support, a shared improvement vision and a supplier link. Our findings question the universality of success factors such as strategic involvement and indicate the need for a more dynamic model of lean implementation.     

Detection and analysis of triterpenic compounds in apple extracts

A new technique for the rapid detection and analysis of triterpenic compounds in apple extracts using HPLC was developed and validated. The main advantage of this technique is the short duration of the analysis – this makes this technique superior to others currently applied for the routine HPLC analysis of triterpenic compounds. The developed, optimized, and validated technique was used for the evaluation of triterpenic compounds in samples of different cultivars of apples, their peels, and flesh. In total, four triterpenic compounds were isolated and identified. Ursolic acid was the dominant compound in all the tested apple samples. The highest amounts of triterpenic compounds were detected in the peels of the ‘Lodel’ apple cultivar, and thus apples of this cultivar may be potentially useful for the isolation of individual compounds and the production of functional food and dietary supplements.     

Bovine milk oligosaccharides decrease gut permeability and improve inflammation and microbial dysbiosis in diet-induced obese mice

Obesity is characterized by altered gut homeostasis, including dysbiosis and increased gut permeability closely linked to the development of metabolic disorders. Milk oligosaccharides are complex sugars that selectively enhance the growth of specific beneficial bacteria in the gastrointestinal tract and could be used as prebiotics. The aim of the study was to demonstrate the effects of bovine milk oligosaccharides (BMO) and Bifidobacterium longum ssp. infantis (B. infantis) on restoring diet-induced obesity intestinal microbiota and barrier function defects in mice. Male C57/BL6 mice were fed a Western diet (WD, 40% fat/kcal) or normal chow (C, 14% fat/kcal) for 7 wk. During the final 2 wk of the study, the diet of a subgroup of WD-fed mice was supplemented with BMO (7% wt/wt). Weekly gavage of B. infantis was performed in all mice starting at wk 3, yet B. infantis could not be detected in any luminal contents when mice were killed. Supplementation of the WD with BMO normalized the cecal and colonic microbiota with increased abundance of Lactobacillus compared with both WD and C mice and restoration of Allobaculum and Ruminococcus levels to that of C mice. The BMO supplementation reduced WD-induced increase in paracellular and transcellular flux in the large intestine as well as mRNA levels of the inflammatory marker tumor necrosis factor α. In conclusion, BMO are promising prebiotics to modulate gut microbiota and intestinal barrier function for enhanced health.     

Freshness Assessment of Thawed and Chilled Cod Fillets Packed in Modified Atmosphere Using Near-infrared Spectroscopy

Near-infrared reflectance (NIR) spectra was recorded of 105 samples of cod mince prepared from chill stored thawed cod fillets of varying quality in modified atmosphere packaging (MAP). Traditional chemical, physical, microbiological and sensory quality methods developed for assessing fresh fish products were determined on the same cod fillets. The purpose was to evaluate the potential of NIR spectroscopy for estimating (i) frozen storage temperature, (ii) frozen storage period and (iii) chill storage period of thawed-chilled MAP Barents Sea cod fillets. Furthermore, the potential for measuring of selected quality attributes as drip loss, water holding capacity and content of dimethylamine by NIR was evaluated. The results of the investigation were presented using multivariate modelling methods such as partial least-squares regression (PLSR) and discriminant partial least-squares regression (DPLSR). Systematic differences in the NIR measurements on minced cod fillets were primarily due to the chill storage duration (days at 2°C) on thawed-chilled MAP fillets. PLSR models based on wavelengths selected by a new Jack-knife method resulted in a correlation coefficient of 0.90 between measured and predicted duration of chill storage period (days at 2°C). The root-mean-square error of cross-validation (RMSECV) was 3.4 d at 2°C. NIR measurements provided promising results for evaluation of freshness for thawed-chilled MAP cod fillets completing the traditionally quality methods. However, it is necessary to study the effect of e.g. sample preparation, season, fishing ground and cod size together with more sophisticated pre-treatments of NIR spectra before the NIR method can be integrated as a method for evaluation of thawed-chilled MAP cod fillets.     

Inactivation of Escherichia coli, Listeria innocua and Saccharomyces cerevisiae in relation to membrane permeabilization and subsequent leakage of intracellular compounds due to pulsed electric field processing

Membrane permeabilization, caused by pulsed electric field (PEF) processing of microbial cells, was investigated by measurement of propidium iodide (PI) uptake with flow cytometry. Inactivation of Escherichia coli, Listeria innocua and Saccharomyces cerevisiae was determined by viable counts, and leakage of intracellular compounds, such as ATP and UV-absorbing substances, was measured in the extracellular environment. Electrical field strength and pulse duration influenced membrane permeabilization of all three tested organisms of which S. cerevisiae was the most PEF sensitive, followed by E. coli and L. innocua. It was shown by viable counts, PI uptake and leakage of intracellular compounds that L. innocua was the most resistant. Increased inactivation corresponded to greater numbers of permeabilized cells, which were reflected by increased PI uptake and larger amounts of intracellular compounds leaking from cells. For E. coli and L. innocua, a linear relationship was observed between the number of inactivated cells (determined as CFU) and cells with permeated membranes (determined by PI uptake), with higher number of inactivated cells than permeated cells. Increased leakage of intracellular compounds with increasing treatment severity provided further evidence that cells were permeabilized. For S. cerevisiae, there was higher PI uptake after PEF treatments, although very little or no inactivation was observed. Results suggest that E. coli and L. innocua cells, which took up PI, lost their ability to multiply, whereas cells of S. cerevisiae, which also took up PI, were not necessarily lethally permeabilized.     

The Antimicrobial Effect of Spice‐Based Marinades against Campylobacter jejuni on Contaminated Fresh Broiler Wings

The antimicrobial effect of spice‐based marinades against Campylobacter jejuni on inoculated fresh broiler wings was investigated. Experiments were carried out with 1 strain of C. jejuni and 6 marinades. Four experimental marinades were composed for the study and contained spices (thyme, rosemary, basil, marjoram, and so on) and different combination of bioactive compounds. Two marinades were commercial and contained spices (black pepper, sweet red pepper, and so on) and chemical additives (monosodium glutamate, sodium diacetate, calcium lactate), 1 commercial marinade was also enriched with bioactive compounds (linalool, cinnamaldehyde, lactic acid). Total aerobic bacterial count was examined to estimate the possible effect of tested marinades on the shelf‐life of marinated broiler wings. Study revealed that thyme‐based marinade was the most effective against C. jejuni on broiler wings and reduced the numbers of campylobacters by 1.04 log colony forming unit (CFU)/g (P ≤ 0.05) during storage for 168 h at 4 °C temperature. Moreover, it was more effective against C. jejuni than commercial marinade with 0.47 log CFU/g (P ≤ 0.05) reduction effect. Both experimental and commercial marinades had very similar effect on the total aerobic bacterial count. Although experimental and commercial marinades had different effect on pH of broiler wings, this parameter did not show a major impact on the antimicrobial effect of tested marinades (P ≥ 0.05). Our study shows that experimental natural thyme‐based marinade can reduce numbers of C. jejuni more effectively than tested commercial marinades.