Alexandros Ch. Stratakos

Industrial scale microwave processing of tomato juice using a novel continuous microwave system

This study evaluated the effect of an industrial scale continuous flow microwave volumetric heating system in comparison to conventional commercial scale pasteurisation for the processing of tomato juice in terms of physicochemical properties, microbial characteristics and antioxidant capacity. The effect against oxidative stress in Caco-2 cells, after in vitro digestion was also investigated. Physicochemical and colour characteristics of juices were very similar between technologies and during storage. Both conventional and microwave pasteurisation inactivated microorganisms and kept them in low levels throughout storage. ABTS[Symbol: see text](+) values, but not ORAC, were higher for the microwave pasteurised juice at day 0 however no significant differences between juices were observed during storage. Juice processed with the microwave system showed an increased cytoprotective effect against H2O2 induced oxidation in Caco-2 cells. Organoleptic analysis revealed that the two tomato juices were very similar. The continuous microwave volumetric heating system appears to be a viable alternative to conventional pasteurisation.

Enhancing the lycopene in vitro bioaccessibility of tomato juice synergistically applying thermal and non-thermal processing technologies

The influence of moderate intensity pulsed electric field pre-processing on increasing the lycopene bioaccessibility of tomato fruit, and the combined effect of blanching, ultrasonic and high intensity pulsed electric field processing on further enhancement of the lycopene bioaccessibility after juicing were investigated. Maximum total lycopene bioaccessibility (9.6%) of the tomato fruit was achieved by a 4μs pre-processed treatment after 24h holding period and further processing results revealed that all treatments were effective to increase the total lycopene. Most of juice processing treatments decreased the release of lycopene from the tomato matrix during digestion. Only the treatment of blanching followed by high intensity pulsed electric field showed a significant release of trans-(4.01±0.48) and cis-(5.04±0.26μg/g) lycopene, achieving 15.6% total lycopene bioaccessibility. Thus, processing of pre-blanched juice using high intensity pulsed electric field, derived from pre-processed tomato was the best overall process to achieve the highest nutritive value.

The STARTEC Decision Support Tool for Better Tradeoffs between Food Safety, Quality, Nutrition, and Costs in Production of Advanced Ready-to-Eat Foods:

A prototype decision support IT-tool for the food industry was developed in the STARTEC project. Typical processes and decision steps were mapped using real life production scenarios of participating food companies manufacturing complex ready-to-eat foods. Companies looked for a more integrated approach when making food safety decisions that would align with existing HACCP systems. The tool was designed with shelf life assessments and data on safety, quality, and costs, using a pasta salad meal as a case product. The process flow chart was used as starting point, with simulation options at each process step. Key parameters like pH, water activity, costs of ingredients and salaries, and default models for calculations of Listeria monocytogenes, quality scores, and vitamin C, were placed in an interactive database. Customization of the models and settings was possible on the user-interface. The simulation module outputs were provided as detailed curves or categorized as “good”; “sufficient”; or “corrective action needed” based on threshold limit values set by the user. Possible corrective actions were suggested by the system. The tool was tested and approved by end-users based on selected ready-to-eat food products. Compared to other decision support tools, the STARTEC-tool is product-specific and multidisciplinary and includes interpretation and targeted recommendations for end-users.

Methods for extracting essential oils

Abstract Essential oils are composite mixtures of volatile compounds most frequently present at low concentrations in plants. Several different extraction techniques are widely employed for the extraction of essential oils such as steam distillation and solvent extraction. These methods are characterized by drawbacks such as low extraction efficiency and selectivity, use of large amounts of solvents, and long extraction times. In many cases, the quality of the essential oil obtained by conventional methods can be influenced by hydrolyzation or oxidation than can take place due to long extraction time and/or high water quantity. Due to these limitations, alternative methods for the extraction of essential oils have been developed which can typically overcome these problems. Supercritical fluid extraction, microwave assisted extraction and ultrasound are novel methods that are now recognized as efficient extraction methods and can significantly reduce extraction times, enhance yields, and quality of essential oil. Although these methods are predominantly exploited on the laboratory scale, they have also found industrial applications, although in most cases to a limited extent.

The in vitro and ex vivo effect of Auranta 3001 in preventing Cryptosporidium hominis and Cryptosporidium parvum infection

BackgroundCryptosporidium is a major cause of diarrhea worldwide in both humans and farm animals with no completely effective treatment available at present. In this study, we assessed the inhibitory effect of different concentrations of Auranta 3001 (0.1, 0.5 and 1%), a novel natural feed supplement, on C. hominis and C. parvum invasion of human ileocecal adenocarcinoma (HCT-8), bovine primary cells and C. parvum invasion of HCT-8, bovine primary cells and bovine intestinal biopsies. The effect of the feed supplement on the production of pro-inflammatory cytokines IL-8 and INF-γ, the anti-inflammatory cytokine IL-10, the expression of CpSUB1 protease gene during infection was also assessed by quantitative PCR (q-PCR). Transepithelial electrical resistance (TEER) was employed to measure the integrity of tight junction dynamics of the culture models.ResultsPre-treatment of intestinal cells or oocysts with the Auranta 3001 significantly reduced the invasiveness of C. hominis and C. parvum against HCT-8 and bovine primary cells in a dose dependent manner. The most pronounced reduction in the invasiveness of both parasites was observed when Auranta 3001 was present during infection. Levels of IL-8 were significantly reduced in both HCT-8 and bovine primary cells, while the levels of INF-γ and IL-10 showed opposite trends in the two cell lines during infection in the presence of Auranta 3001. CpSUB1 gene protease expression, which mediates infection, was significantly reduced suggesting that this enzyme is a possible target of Auranta 3001.ConclusionsAlthough, C. hominis and C. parvum use different invasion mechanisms to infect cells, the novel feed additive can significantly attenuate the entry of Cryptosporidium in HCT-8 cells, primary bovine cells and bovine intestinal biopsies and thus provide an alternative method to control cryptosporidiosis.

A Review of the Effect of Management Practices on Campylobacter Prevalence in Poultry Farms

Poultry is frequently associated with campylobacteriosis in humans, with Campylobacter jejuni being the most usual Campylobacter associated with disease in humans. Far-reaching research on Campylobacter was undertaken over the past two decades. This has resulted in interventions being put in place on farms and in processing plants. Despite these interventions, coupled with increased media coverage to educate the consumer on Campylobacter prevalence and campylobacteriosis, human health incidents are still high. Recent research is now shifting toward further understanding of the microorganisms to challenge interventions in place and to look at further and more relevant interventions for the reduction in human incidents. Farm practices play a key role in the control of colonization within poultry houses and among flocks. Prevalence at the farm level can be up to 100% and time of colonization may vary widely between flocks. Considerable research has been performed to understand how farm management and animal health practices can affect colonization on farms. This review will focus on farm practices to date as a baseline for future interventions as the microorganism becomes better understood. Further research is required to understand the chicken microbiome and factors influencing vertical transmission. The persistence of Campylobacter in animal and environmental reservoirs within and around farms requires further investigation to tailor farm practices toward preventing such reservoirs. IMPLICATIONS This review gives an overview of farm practices and their effect on Campylobacter prevalence in poultry. Various elements of farm practices have been captured in this review.

A Novel Natural Antimicrobial Can Reduce the in vitro and in vivo Pathogenicity of T6SS Positive Campylobacter jejuni and Campylobacter coli Chicken Isolates

Human campylobacteriosis is considered one of the most common foodborne diseases worldwide with poultry identified as the main source of infection accounting for 50–80% of human cases. Highly virulent Campylobacter spp., positive for the Type VI secretion system (T6SS), which have an increased ability to adhere to and invade the host gastrointestinal epithelium are highly prevalent in poultry. Multidrug resistant strains of bacteria are rapidly evolving and therefore, new antimicrobials to supplement animal feed that are able to control Campylobacter species, are in great need. The work presented herein indicates that a novel phenolic antimicrobial, Auranta 3001, is able to reduce the adhesion and invasion of human intestinal epithelial cells (HCT-8) by two T6SS positive chicken isolates, C. jejuni RC039 (p < 0.05) and C. coli RC013 (p < 0.001). Exposure of C. jejuni RC039 and C. coli RC013 to Auranta 3001 downregulated the expression of hcp and cetB genes, known to be important in the functionality of T6SS. Furthermore, the reduced adhesion and invasion is associated with a significant decrease in bacterial motility of both isolates (p < 0.05–p < 0.001) in vitro. Most importantly our in vivo results show that Auranta 3001 is able to reduce cecum colonization levels from log 8 CFU/ml to log 2 CFU/ml for C. jejuni RC039 and from log 7 CFU/ml to log 2 CFU/ml for C. coli RC013. In conclusion, this novel antimicrobial is able to reduce the pathogenic properties of T6SS campylobacters in vitro and also to decrease colonization in vivo.

Comprehensive Longitudinal Microbiome Analysis of the Chicken Cecum Reveals a Shift From Competitive to Environmental Drivers and a Window of Opportunity for Campylobacter

Chickens are a key food source for humans yet their microbiome contains bacteria that can be pathogenic to humans, and indeed potentially to chickens themselves. Campylobacter is present within the chicken gut and is the leading cause of bacterial foodborne gastroenteritis within humans worldwide. Infection can lead to secondary sequelae such as Guillain-Barré syndrome and stunted growth in children from low-resource areas. Despite the global health impact and economic burden of Campylobacter, how and when Campylobacter appears within chickens remains unclear. The lack of day to day microbiome data with replicates, relevant metadata, and a lack of natural infection studies have delayed our understanding of the chicken gut microbiome and Campylobacter. Here, we performed a comprehensive day to day microbiome analysis of the chicken cecum from day 3 to 35 (12 replicates each day; final n = 379). We combined metadata such as chicken weight and feed conversion rates to investigate what the driving forces are for the microbial changes within the chicken gut over time, and how this relates to Campylobacter appearance within a natural habitat setting. We found a rapidly increasing microbial diversity up to day 12 with variation observed both in terms of genera and abundance, before a stabilization of the microbial diversity after day 20. In particular, we identified a shift from competitive to environmental drivers of microbial community from days 12 to 20 creating a window of opportunity whereby Campylobacter can appear. Campylobacter was identified at day 16 which was 1 day after the most substantial changes in metabolic profiles observed. In addition, microbial variation over time is most likely influenced by the diet of the chickens whereby significant shifts in OTU abundances and beta dispersion of samples often corresponded with changes in feed. This study is unique in comparison to the most recent studies as neither sampling was sporadic nor Campylobacter was artificially introduced, thus the experiments were performed in a natural setting. We believe that our findings can be useful for future intervention strategies and help reduce the burden of Campylobacter within the food chain.

The antimicrobial effect of a commercial mixture of natural antimicrobials against Escherichia coli O157:H7

Ruminants are important reservoirs of E. coli O157:H7 and are considered as the major source of most foodborne outbreaks (e.g., 2017 outbreak in Germany, 2014 and 2016 outbreaks in United States, all linked to beef products). A promising strategy to reduce E. coli O157 is using antimicrobials to reduce the pathogen levels and/or virulence within the animal gastrointestinal tract and thus foodborne disease. The aim of the study was to determine the efficacy of a commercial mixture of natural antimicrobials against E. coli O157. The minimum inhibitory concentration and minimum bactericidal concentration of the antimicrobial were quantitatively determined and found to be 0.5% and 0.75% (v/v) of the natural antimicrobial, respectively. Microbial growth kinetics was also used to determine the effect of the antimicrobial on the pathogen. The natural antimicrobial affected the cell membrane of E. coli O157, as demonstrated by the increase in relative electric conductivity and increase in protein and nucleic acid release. The antimicrobial was also able to significantly reduce the concentration on E. coli O157 in a model rumen system. Biofilm assays showed that subinhibitory concentrations of the antimicrobial significantly reduced the E. coli 0157 biofilm forming capacity without influencing pathogen growth. In addition, the natural antimicrobial was able to reduce motility and exopolysaccharide production. Subinhibitory concentrations of the antimicrobial had no effect on AI-2 production. These findings suggest that the natural antimicrobial exerts an antimicrobial effect against E. coli O157 in vitro and in a model rumen system and could be potentially used to control this pathogen in the animal gut. The results also indicate that subinhibitory concentrations of the antimicrobial effectively reduce biofilm formation, motility, and exopolysaccharide production.

Evaluation of the efficacy of multiple physical, biological and natural antimicrobial interventions for control of pathogenic Escherichia coli on beef

Antimicrobial effects of multiple physical, biological and natural interventions on pathogenic Escherichia coli in raw beef were assessed. A cocktail of E. coli strains was inoculated onto gamma-irradiated beef and enumerated immediately after each intervention and during storage at 4 °C for 7 days. Of the physical interventions, silver-containing antimicrobial packaging and ozone gas treatment did not show significant antimicrobial effects, however cold plasma treatment reduced E. coli levels by 0.9 and 1.82 log10 CFU/cm2 after 2 and 5 min treatments, respectively. A phage cocktail reduced E. coli counts by 0.63 and 1.16 log10 CFU/g after 24 h storage at 4 and 12 °C, respectively. Of the natural interventions, vinegar and lactic acid (5%) washes for 5 min caused reductions of ∼1 log10 CFU/g immediately after treatment, whereas lactoferrin and nisin treatments, separately or in combination, had insignificant antimicrobial effects. Nanoemulsions containing carvacrol or thyme essential oils caused immediate E. coli reductions of 1.41 and 1.36 log10 CFU/g, respectively, plus a progressive reduction in viable numbers during storage at 4 °C. Our findings suggest that cold plasma, bacteriophages, vinegar, lactic acid, or carvacrol and thyme essential oil nanoemulsions could potentially be of use to the beef industry for controlling pathogenic E. coli contamination.