The fourth industrial revolution in the food industry-Part I: Industry 4.0 technologies.

Climate change, the growth in world population, high levels of food waste and food loss, and the risk of new disease or pandemic outbreaks are examples of the many challenges that threaten future food sustainability and the security of the planet and urgently need to be addressed. The fourth industrial revolution, or Industry 4.0, has been gaining momentum since 2015, being a significant driver for sustainable development and a successful catalyst to tackle critical global challenges. This review paper summarizes the most relevant food Industry 4.0 technologies including, among others, digital technologies (e.g., artificial intelligence, big data analytics, Internet of Things, and blockchain) and other technological advances (e.g., smart sensors, robotics, digital twins, and cyber-physical systems). Moreover, insights into the new food trends (such as 3D printed foods) that have emerged as a result of the Industry 4.0 technological revolution will also be discussed in Part II of this work. The Industry 4.0 technologies have significantly modified the food industry and led to substantial consequences for the environment, economics, and human health. Despite the importance of each of the technologies mentioned above, ground-breaking sustainable solutions could only emerge by combining many technologies simultaneously. The Food Industry 4.0 era has been characterized by new challenges, opportunities, and trends that have reshaped current strategies and prospects for food production and consumption patterns, paving the way for the move toward Industry 5.0.

Prebiotic effect of inulin-type fructans on faecal microbiota and short-chain fatty acids in type 2 diabetes: a randomised controlled trial.

PURPOSE: Compared to a healthy population, the gut microbiota in type 2 diabetes presents with several unfavourable features that may impair glucose regulation. The aim of this study was to evaluate the prebiotic effect of inulin-type fructans on the faecal microbiota and short-chain fatty acids (SCFA) in patients with type 2 diabetes. METHODS: The study was a placebo controlled crossover study, where 25 patients (15 men) aged 41-71 years consumed 16 g of inulin-type fructans (a mixture of oligofructose and inulin) and 16-g placebo (maltodextrin) for 6 weeks in randomised order. A 4-week washout separated the 6 weeks treatments. The faecal microbiota was analysed by high-throughput 16S rRNA amplicon sequencing and SCFA in faeces were analysed using vacuum distillation followed by gas chromatography. RESULTS: Treatment with inulin-type fructans induced moderate changes in the faecal microbiota composition (1.5%, p = 0.045). A bifidogenic effect was most prominent, with highest positive effect on operational taxonomic units (OTUs) of Bifidobacterium adolescentis, followed by OTUs of Bacteroides. Significantly higher faecal concentrations of total SCFA, acetic acid and propionic acid were detected after prebiotic consumption compared to placebo. The prebiotic fibre had no effects on the concentration of butyric acid or on the overall microbial diversity. CONCLUSION: Six weeks supplementation with inulin-type fructans had a significant bifidogenic effect and induced increased concentrations of faecal SCFA, without changing faecal microbial diversity. Our findings suggest a moderate potential of inulin-type fructans to improve gut microbiota composition and to increase microbial fermentation in type 2 diabetes. TRIAL REGISTRATION: The trial is registered at clinicaltrials.gov (NCT02569684).

Correction to: Prebiotic effect of inulin­type fructans on faecal microbiota and short­chain fatty acids in type 2 diabetes: a randomised controlled trial.

The original version of this article unfortunately contained a mistake. The presentation of Fig. 4 was incorrect.

Microbiota formed on attached stainless steel coupons correlates with the natural biofilm of the sink surface in domestic kitchens.

Stainless steel coupons are frequently used in biofilm studies in the laboratory, as this material is commonly used in the food industry. The coupons are attached to different surfaces to create a "natural" biofilm to be studied further in laboratory trials. However, little has been done to investigate how well the microbiota on such coupons represents the surrounding environment. The microbiota on sink wall surfaces and on new stainless steel coupons attached to the sink wall for 3 months in 8 domestic kitchen sinks was investigated by next-generation sequencing (MiSeq) of the 16S rRNA gene derived from DNA and RNA (cDNA), and by plating and identification of colonies. The mean number of colony-forming units was about 10-fold higher for coupons than sink surfaces, and more variation in bacterial counts between kitchens was seen on sink surfaces than coupons. The microbiota in the majority of biofilms was dominated by Moraxellaceae (genus Moraxella/Enhydrobacter) and Micrococcaceae (genus Kocuria). The results demonstrated that the variation in the microbiota was mainly due to differences between kitchens (38.2%), followed by the different nucleic acid template (DNA vs RNA) (10.8%), and that only 5.1% of the variation was a result of differences between coupons and sink surfaces. The microbiota variation between sink surfaces and coupons was smaller for samples based on their RNA than on their DNA. Overall, our results suggest that new stainless steel coupons are suited to model the dominating part of the natural microbiota of the surrounding environment and, furthermore, are suitable for different downstream studies.

High nutrient availability reduces the diversity and stability of the equine caecal microbiota.

BACKGROUND: It is well known that nutrient availability can alter the gut microbiota composition, while the effect on diversity and temporal stability remains largely unknown. METHODS: Here we address the equine caecal microbiota temporal stability, diversity, and functionality in response to diets with different levels of nutrient availability. Hay (low and slower nutrient availability) versus a mixture of hay and whole oats (high and more rapid nutrient availability) were used as experimental diets. RESULTS: We found major effects on the microbiota despite that the caecal pH was far from sub-clinical acidosis. We found that the low nutrient availability diet was associated with a higher level of both diversity and temporal stability of the caecal microbiota than the high nutrient availability diet. These observations concur with general ecological theories, suggesting a stabilising effect of biological diversity and that high nutrient availability has a destabilising effect through reduced diversity. CONCLUSION: Nutrient availability does not only change the composition but also the ecology of the caecal microbiota.

Diagnosing the cage of covariance to increase understanding and robustness of spectroscopic calibration models.

When vibrational spectroscopy is used for quantification purposes, multivariate analysis is often used to extract information from covariances between the spectra and any given reference values. In complex samples, there is a high risk that the constituents covary with each other. In such scenarios many methods may confuse the analytes and use signal from several analytes, rather than just the analyte of interest. While this allows the method to use more signal, and thus have a better effective signal-to-noise ratio, it also makes them less robust to changes to the chemical composition in the samples. This effect has been termed the cage of covariance. In order to avoid cage of covariance to affect predictive performances, it is highly important to have simple diagnostic tools to analyze and review this effect. Therefore, in the present paper, a systematic overview of tools for diagnosing and quantifying the cage of covariance in spectroscopic calibration models is provided. A collection of previously published methods with some expansions is provided, as well as two completely new tools: covariance ratio and virtual spiking. Practical applications of the tools on three different datasets are also shown.

A portable dry film FTIR instrument for industrial food and bioprocess applications.

The main objective of this study was to design, build, and test a compact, multi-well, portable dry film FTIR system for industrial food and bioprocess applications. The system features dry film sampling on a circular rotating disc comprising 31 wells, a design that was chosen to simplify potential automation and robotic sample handling at a later stage. Calibration models for average molecular weight (AMW, 200 samples) and collagen content (68 samples) were developed from the measurements of industrially produced protein hydrolysate samples in a controlled laboratory environment. Similarly, calibration models for the prediction of lactate content in samples from cultivation media (59 samples) were also developed. The portable dry film FTIR system showed reliable model characteristics which were benchmarked with a benchtop FTIR system. Subsequently, the portable dry film FTIR system was deployed in a bioprocessing plant, and protein hydrolysate samples were measured at-line in an industrial environment. This industrial testing involved building a calibration model for predicting AMW using 60 protein hydrolysate samples measured at-line using the portable dry film FTIR system and subsequent model validation using a test set of 26 samples. The industrial calibration in terms of coefficient of determination (R2 = 0.94), root mean square of cross-validation (RMSECV = 194 g mol-1), and root mean square of prediction (RMSEP = 162 g mol-1) demonstrated low prediction errors as compared to benchtop FTIR measurements, with no statistical difference between the calibration models of the two FTIR systems. This is to the authors' knowledge the first study for developing and employing a portable dry film FTIR system in the enzymatic protein hydrolysis industry for successful at-line measurements of protein hydrolysate samples. The study therefore suggests that the portable dry film FTIR instrument has huge potential for in/at-line applications in the food and bioprocessing industries.

In-Line Near-Infrared Spectroscopy Gives Rapid and Precise Assessment of Product Quality and Reveals Unknown Sources of Variation-A Case Study from Commercial Cheese Production.

Quality testing in the food industry is usually performed by manual sampling and at/off-line laboratory analysis, which is labor intensive, time consuming, and may suffer from sampling bias. For many quality attributes such as fat, water and protein, in-line near-infrared spectroscopy (NIRS) is a viable alternative to grab sampling. The aim of this paper is to document some of the benefits of in-line measurements at the industrial scale, including higher precision of batch estimates and improved process understanding. Specifically, we show how the decomposition of continuous measurements in the frequency domain, using power spectral density (PSD), may give a useful view of the process and serve as a diagnostic tool. The results are based on a case regarding the large-scale production of Gouda-type cheese, where in-line NIRS was implemented to replace traditional laboratory measurements. In conclusion, the PSD of in-line NIR predictions revealed unknown sources of variation in the process that could not have been discovered using grab sampling. PSD also gave the dairy more reliable data on key quality attributes, and laid the foundation for future improvements.

FTIR-based prediction of collagen content in hydrolyzed protein samples.

Fourier transform infrared spectroscopy (FTIR) is a powerful analytical tool that has been used for protein and peptide characterization for decades. In the present study, the objective was to investigate if FTIR can be used to predict collagen content in hydrolyzed protein samples. All samples were obtained from enzymatic protein hydrolysis (EPH) of poultry by-products providing a span in collagen content from 0.3% to 37.9% (dry weight), and the FTIR analysis was performed using dry film FTIR. Since nonlinear effects were revealed by calibration using standard partial least squares (PLS) regression, Hierarchical Cluster-based PLS (HC-PLS) calibration models were constructed. The HC-PLS model provided a low prediction error when validated using an independent test set (RMSE = 3.3% collagen), while validation using real industrial samples also showed satisfying results (RMSE = 3.2%). The results corresponded well with previously published FTIR-based studies of collagen, and characteristic spectral features for collagen were well identified in the regression models. Covariance between collagen content and other EPH related processing parameters could also be ruled out in the regression models. To the authors' knowledge, this is the first time that collagen content has been systematically studied in solutions of hydrolysed proteins using FTIR. This is also one of few examples where FTIR is successfully used to quantify protein composition. The dry-film FTIR approach presented in the study is expected to be an important tool in the growing industrial segment that is based on sustainable utilization of collagen-rich biomass.

The role of biospectroscopy and chemometrics as enabling technologies for upcycling of raw materials from the food industry.

It is important to utilize the entire animal in meat and fish production to ensure sustainability. Rest raw materials, such as bones, heads, trimmings, and skin, contain essential nutrients that can be transformed into high-value products. Enzymatic protein hydrolysis (EPH) is a bioprocess that can upcycle these materials to create valuable proteins and fats. This paper focuses on the role of spectroscopy and chemometrics in characterizing the quality of the resulting protein product and understanding how raw material quality and processing affect it. The article presents recent developments in chemical characterisation and process modelling, with a focus on rest raw materials from poultry and salmon production. Even if some of the technology is relatively mature and implemented in many laboratories and industries, there are still open challenges and research questions. The main challenges are related to the transition of technology and insights from laboratory to industrial scale, and the link between peptide composition and critical product quality attributes.

Dietary intake of micronized avian eggshell membrane in aged mice reduces circulating inflammatory markers, increases microbiota diversity, and attenuates skeletal muscle aging.

Introduction: Avian eggshell membrane (ESM) is a complex extracellular matrix comprising collagens, glycoproteins, proteoglycans, and hyaluronic acid. We have previously demonstrated that ESM possesses anti-inflammatory properties in vitro and regulates wound healing processes in vivo. The present study aimed to investigate if oral intake of micronized ESM could attenuate skeletal muscle aging associated with beneficial alterations in gut microbiota profile and reduced inflammation. Methods: Elderly male C57BL/6 mice were fed an AIN93G diet supplemented with 0, 0.1, 1, or 8% ESM. Young mice were used as reference. The digestibility of ESM was investigated using the static in vitro digestion model INFOGEST for older people and adults, and the gut microbiota profile was analyzed in mice. In addition, we performed a small-scale pre-clinical human study with healthy home-dwelling elderly (>70 years) who received capsules with a placebo or 500 mg ESM every day for 4 weeks and studied the effect on circulating inflammatory markers. Results and discussion: Intake of ESM in elderly mice impacted and attenuated several well-known hallmarks of aging, such as a reduction in the number of skeletal muscle fibers, the appearance of centronucleated fibers, a decrease in type IIa/IIx fiber type proportion, reduced gene expression of satellite cell markers Sdc3 and Pax7 and increased gene expression of the muscle atrophy marker Fbxo32. Similarly, a transition toward the phenotypic characteristics of young mice was observed for several proteins involved in cellular processes and metabolism. The digestibility of ESM was poor, especially for the elderly condition. Furthermore, our experiments showed that mice fed with 8% ESM had increased gut microbiota diversity and altered microbiota composition compared with the other groups. ESM in the diet also lowered the expression of the inflammation marker TNFA in mice and in vitro in THP-1 macrophages. In the human study, intake of ESM capsules significantly reduced the inflammatory marker CRP. Altogether, our results suggest that ESM, a natural extracellular biomaterial, may be attractive as a nutraceutical candidate with a possible effect on skeletal muscle aging possibly through its immunomodulating effect or gut microbiota.

A Three-Day Intervention With Granola Containing Cereal Beta-Glucan Improves Glycemic Response and Changes the Gut Microbiota in Healthy Individuals: A Crossover Study.

Intake of soluble fibers including beta-glucan, is known to improve post-prandial glycemic response. The mechanisms have been attributed to the viscous gel forming in the stomach and small intestine, giving a longer absorption time. However, recent evidence suggests a link between intake of beta-glucan and improved glycemic regulation at subsequent meals through the gut microbiota. We investigated the short-term effect of granola with different amounts of cereal beta-glucan on glycemic response and gut microbiota. After a two-week run-in period (baseline), fourteen healthy, normal weight adults completed a dose-response dietary crossover study. Different amounts of cereal beta-glucan (low: 0.8 g, medium: 3.2 g and high: 6.6 g) were provided in granola and eaten with 200 ml low-fat milk as an evening meal for three consecutive days. Blood glucose and insulin were measured fasted and after an oral glucose tolerance test (OGTT) the following day, in addition to peptide YY (PYY) and glucagon-like peptide (GLP-2), fasting short chain fatty acids (SCFA) in blood, breath H2, and gut microbiota in feces. Only the intervention with medium amounts of beta-glucan decreased blood glucose and insulin during OGTT compared to baseline. Fasting PYY increased with both medium and high beta-glucan meal compared to the low beta-glucan meal. The microbiota and SCFAs changed after all three interventions compared to baseline, where acetate and butyrate increased, while propionate was unchanged. Highest positive effect size after intake of beta-glucan was found with Haemophilus, followed by Veillonella and Sutterella. Furthermore, we found several correlations between different bacterial taxa and markers of glycemic response. In summary, intake of granola containing 3.2 g cereal beta-glucan as an evening meal for three consecutive days reduced the glycemic response after an OGTT 0-180 min and changed gut microbiota composition. Since we cannot rule out that other fiber types have contributed to the effect, more studies are needed to further explore the effect of cereal beta-glucan on glycemic regulation. Clinical Trial Registration: [www.clinicaltrials.gov], identifier [NCT03293693].

Analysing microbiome intervention design studies: Comparison of alternative multivariate statistical methods.

The diet plays a major role in shaping gut microbiome composition and function in both humans and animals, and dietary intervention trials are often used to investigate and understand these effects. A plethora of statistical methods for analysing the differential abundance of microbial taxa exists, and new methods are constantly being developed, but there is a lack of benchmarking studies and clear consensus on the best multivariate statistical practices. This makes it hard for a biologist to decide which method to use. We compared the outcomes of generic multivariate ANOVA (ASCA and FFMANOVA) against statistical methods commonly used for community analyses (PERMANOVA and SIMPER) and methods designed for analysis of count data from high-throughput sequencing experiments (ALDEx2, ANCOM and DESeq2). The comparison is based on both simulated data and five published dietary intervention trials representing different subjects and study designs. We found that the methods testing differences at the community level were in agreement regarding both effect size and statistical significance. However, the methods that provided ranking and identification of differentially abundant operational taxonomic units (OTUs) gave incongruent results, implying that the choice of method is likely to influence the biological interpretations. The generic multivariate ANOVA tools have the flexibility needed for analysing multifactorial experiments and provide outputs at both the community and OTU levels; good performance in the simulation studies suggests that these statistical tools are also suitable for microbiome data sets.

Sensory Assessment of Fish and Chicken Protein Hydrolysates. Evaluation of NMR Metabolomics Profiling as a New Prediction Tool.

Nuclear magnetic resonance (NMR) metabolomics profiling was evaluated as a new tool in sensory assessment of protein hydrolysates. Hydrolysates were produced on the basis of different raw materials (cod, salmon, and chicken), enzymes (Food Pro PNL and Bromelain), and hydrolysis time (10 and 50 min). The influence of raw material and hydrolysis parameters on sensory attributes was determined by traditional descriptive sensory analysis and 1H NMR spectroscopy. The raw material had a major influence on the attribute intensity and metabolite variation, followed by enzyme and hydrolysis time. However, the formation of bitter taste was not affected by the raw material. Partial least-squares regression (PLSR) on 1H NMR and sensory data provided good models (Q2 = 0.55-0.89) for 11 of the 17 evaluated attributes, including bitterness. Significant metabolite-attribute associations were identified. The study confirms the potential prediction of the sensory properties of protein hydrolysates from cod, salmon, and chicken based on 1H NMR metabolomics profiling.

Fraud in Animal Origin Food Products: Advances in Emerging Spectroscopic Detection Methods over the Past Five Years.

Animal origin food products, including fish and seafood, meat and poultry, milk and dairy foods, and other related products play significant roles in human nutrition. However, fraud in this food sector frequently occurs, leading to negative economic impacts on consumers and potential risks to public health and the environment. Therefore, the development of analytical techniques that can rapidly detect fraud and verify the authenticity of such products is of paramount importance. Traditionally, a wide variety of targeted approaches, such as chemical, chromatographic, molecular, and protein-based techniques, among others, have been frequently used to identify animal species, production methods, provenance, and processing of food products. Although these conventional methods are accurate and reliable, they are destructive, time-consuming, and can only be employed at the laboratory scale. On the contrary, alternative methods based mainly on spectroscopy have emerged in recent years as invaluable tools to overcome most of the limitations associated with traditional measurements. The number of scientific studies reporting on various authenticity issues investigated by vibrational spectroscopy, nuclear magnetic resonance, and fluorescence spectroscopy has increased substantially over the past few years, indicating the tremendous potential of these techniques in the fight against food fraud. It is the aim of the present manuscript to review the state-of-the-art research advances since 2015 regarding the use of analytical methods applied to detect fraud in food products of animal origin, with particular attention paid to spectroscopic measurements coupled with chemometric analysis. The opportunities and challenges surrounding the use of spectroscopic techniques and possible future directions will also be discussed.

Near-infrared spectroscopy detects woody breast syndrome in chicken fillets by the markers protein content and degree of water binding.

The muscle syndrome woody breast (WB) impairs quality of chicken fillets and is a challenge to the poultry meat industry. There is a need for online detection of affected fillets for automatic quality sorting in process. Near-infrared spectroscopy (NIRS) is a promising method, and in this study we elucidate the spectral properties of WB versus normal fillets. On a training set of 50 chicken fillets (20 normal, 30 WB), we measured NIR, nuclear magnetic resonance (NMR) T2 relaxation distributions, and crude chemical composition. NIRS could estimate protein in the fillets with an accuracy of ±0.64 percentage points. T2 distributions showed that there was a larger share of free water in WB fillets. This difference in water binding generated a shift and narrowing of the water absorption peak in NIR around 980 nm, quantified by a bound water index (BWI). The correlation between BWI and T2 distributions was 0.78, indicating that NIRS contains information about degree of water binding. Discriminant analysis showed that NIRS obtained 100% correct classification of normal versus WB on the training set, and 96% correct classification on a test set of 52 fillets. The main reason for why NIRS can successfully discriminate between WB and normal fillets is the methods sensitivity to both protein content and degree of water binding in the muscle, both established markers for WB. The classification model can be based on NIR spectra only, calibration against protein is not needed. The affected muscle tissue associated with the WB syndrome is unevenly distributed in the fillets, and this heterogeneity was characterized by NIRS and NMR. Clear differences in water binding properties were found between the superficial 1 cm layer and the deeper layer at 1 to 2 cm depth. Significant differences in protein estimates by NIRS at different measurement points along the chicken fillets were obtained for WB fillets. The findings suggest how to obtain optimal sampling with NIRS for best possible discrimination between WB and normal breast fillets.

Determination of critical levels of residual oxygen to minimize discoloration of sliced packaged Norwegian salami under light display.

Discoloration of sliced packaged salami is contributing to rejection of the product, food waste and economical loss. A combination of residual O2 in the headspace of packages and light is causing photooxidation and deterioration of colour. The aim of this study was to establish maximum tolerable concentrations of residual O2 in packages of salami slices with 100% N2 under light display at 4 and 20°C. Salami sausages had variable inherent O2 consumption rate. Storage of salami in 1% O2 in darkness did not induce discoloration. The upper limits for O2 for avoiding discoloration under light were variable in the range 0.1-1.0%, depending on temperature and type of salami. Display at 20°C increased the rate of O2 depletion compared to 4°C. To minimize discoloration, sliced and packaged salami should be stored in darkness at approximately 20°C until the level of residual O2 is reduced below a critical limit.

Survival of Five Strains of Shiga Toxigenic Escherichia coli in a Sausage Fermentation Model and Subsequent Sensitivity to Stress from Gastric Acid and Intestinal Fluid.

The ability of foodborne pathogens to exhibit adaptive responses to stressful conditions in foods may enhance their survival when passing through the gastrointestinal system. We aimed to determine whether Escherichia coli surviving stresses encountered during a model dry-fermented sausage (DFS) production process exhibit enhanced tolerance and survival in an in vitro gastrointestinal model. Salami sausage batters spiked with five E. coli isolates, including enterohaemorrhagic E. coli strains isolated from different DFS outbreaks, were fermented in a model DFS process (20°C, 21 days). Control batters spiked with the same strains were stored at 4°C for the same period. Samples from matured model sausages and controls were thereafter exposed to an in vitro digestion challenge. Gastric exposure (pH 3) resulted in considerably reduced survival of the E. coli strains that had undergone the model DFS process. This reduction continued after entering intestinal challenge (pH 8), but growth resumed after 120 min. When subjected to gastric challenge for 120 min, E. coli that had undergone the DFS process showed about 2.3 log10⁡ lower survival compared with those kept in sausage batter at 4°C. Our results indicated that E. coli strains surviving a model DFS process exhibited reduced tolerance to subsequent gastric challenge at low pH.

Effect of relevant environmental stresses on survival of enterohemorrhagic Escherichia coli in dry-fermented sausage.

Dry-fermented sausages (DFSs) have been linked to several serious foodborne outbreaks of enterohemorrhagic Escherichia coli (EHEC). The ability of pathogens to utilize adaptive responses to different stressful conditions intended to control their growth in foods, food preparation and production processes may enhance their survival. In certain cases, induced tolerance to one type of stress may lead to enhanced resistance to the applied stress as well as to other stresses. We exposed two EHEC strains, MF3582 of serotype O157:H- and MF5554 of serogroup O145, to different stresses commonly encountered during a production process. The two EHEC strains, previously shown to have different abilities to survive DFS production process conditions, were subjected to low temperatures (4°C and 12°C), 5% NaCl or 1% lactic acid for 6days prior to being added to sausage batters. Survival of EHEC was recorded in salami of two recipes, fermented at two temperatures (20°C and 30°C). The results showed that recipe type had the largest impact on EHEC reductions where Moderate recipe (MR) salami batters containing increased levels of NaCl, glucose and NaNO2 provided enhanced EHEC reductions in salami (2.6 log10) compared to Standard recipe (SR) salami (1.7 log10). Effects of pre-exposure stresses were dependent both on strain and recipe. While acid adaptation of MF5554 provided enhanced log10 reductions from 2.0 to 3.0 in MR sausages, adaptation to a combination of acid and salt stress showed the opposite effect in SR sausages with reductions of only 1.1 log10 as compared to the average of 1.8 log10 for the other SR sausages. Otherwise, the salt and acid adaptation single stresses had relatively small effects on EHEC survival through the DFS production process and subsequent storage and freeze/thaw treatments. Growing cells and cells frozen in batter survived poorly in MR sausages with an average reduction of 3.4 and 3.2 log10, respectively. The reductions of EHEC after storage of DFS increased with higher temperature and storage time. Up to 3.7 log10 additional reduction was obtained when MF3582 was stored for 2months at 20°C. In conclusion, adaptation of EHEC to acid, salt and low temperatures prior to being introduced in a DFS production process has limited, but strain dependent effects on EHEC reductions. Producers should avoid conditions leading to acid and salt adapted cells that can contaminate the sausage batter. Recipe parameters had the largest impact on EHEC reductions while storage at 20°C is effective for enhanced reductions in finished products.

Effect of Dietary Fibers on Cecal Microbiota and Intestinal Tumorigenesis in Azoxymethane Treated A/J Min/+ Mice.

Foods naturally high in dietary fiber are generally considered to protect against development of colorectal cancer (CRC). However, the intrinsic effect of dietary fiber on intestinal carcinogenesis is unclear. We used azoxymethane (AOM) treated A/J Min/+ mice, which developed a significantly higher tumor load in the colon than in the small intestine, to compare the effects of dietary inulin (IN), cellulose (CE) or brewers spent grain (BSG) on intestinal tumorigenesis and cecal microbiota. Each fiber was tested at two dose levels, 5% and 15% (w/w) content of the AIN-93M diet. The microbiota was investigated by next-generation sequencing of the 16S rRNA gene (V4). We found that mice fed IN had approximately 50% lower colonic tumor load than mice fed CE or BSG (p<0.001). Surprisingly, all three types of fiber caused a dose dependent increase of colonic tumor load (p<0.001). The small intestinal tumor load was not affected by the dietary fiber interventions. Mice fed IN had a lower bacterial diversity than mice fed CE or BSG. The Bacteroidetes/Firmicutes ratio was significantly (p = 0.003) different between the three fiber diets with a higher mean value in IN fed mice compared with BSG and CE. We also found a relation between microbiota and the colonic tumor load, where many of the operational taxonomic units (OTUs) related to low tumor load were significantly enriched in mice fed IN. Among the OTUs related to low tumor load were bacteria affiliated with the Bacteroides genus. These results suggest that type of dietary fiber may play a role in the development of CRC, and that the suppressive effect of IN on colonic tumorigenesis is associated with profound changes in the cecal microbiota profile.