A Highly Sensitive Method for the Detection of Hydrolyzed Gluten in Beer Samples Using LFIA.

Most gluten analysis methods have been developed to detect intact gluten, but they have shown limitations in certain foods and beverages in which gluten proteins are hydrolyzed. Methods based on G12/A1 moAbs detect the sequences of gluten immunogenic peptides (GIP), which are the main contributors to the immune response of celiac disease (CD). Immunogenic sequences with tandem epitopes for G12/A1 have been found in beers with <20 mg/kg gluten, which could be consumed by CD patients according to the Codex Alimentarius. Therefore, an accurate method for the estimation of the immunogenicity of a beer is to use two moAbs that can recognize celiac T cell epitopes comprising most of the immunogenic response. Here, a specific and sensitive method based on G12/A1 LFIA was developed to detect GIP in beers labeled gluten-free or with low gluten content, with an LOD of 0.5 mg/kg. A total of 107 beers were analyzed, of those 6.5% showed levels higher than 20 mg/kg gluten and 29% showed levels above the LOD. In addition, G12/A1 LFIA detected gluten in 15 more beer samples than competitive ELISA with another antibody. Despite their labeling, these beers contained GIP which may cause symptoms and/or intestinal damage in CD patients.

Probiotic and Functional Properties of Limosilactobacillus reuteri INIA P572.

Limosilactobacillus reuteri INIA P572 is a strain able to produce the antimicrobial compound reuterin in dairy products, exhibiting a protective effect against some food-borne pathogens. In this study, we investigated some probiotic properties of this strain such as resistance to gastrointestinal passage or to colonic conditions, reuterin production in a colonic environment, and immunomodulatory activity, using different in vitro and in vivo models. The results showed a high resistance of this strain to gastrointestinal conditions, as well as capacity to grow and produce reuterin in a human colonic model. Although the in vitro assays using the RAW 264.7 macrophage cell line did not demonstrate direct immunomodulatory properties, the in vivo assays using a Dextran Sulphate Sodium (DSS)-induced colitic mice model showed clear immunomodulatory and protective effects of this strain.

Optimization of reuterin production in cheese by Lactobacillus reuteri.

Cheeses manufactured from pasteurized milk supplemented with glycerol and reuterin-producing Lactobacillus reuteri INIA P572 as adjunct to the commercial starter culture were analysed in order to optimize a biopreservation strategy. The highest reuterin concentration determined by a colorimetric assay was detected on day 1 in cheeses with 100-500 mM glycerol. The presence of reuterin was confirmed by a direct detection technique as HPLC. Cheeses made with L. reuteri and 200 or 500 mM glycerol showed a red tendency in color in comparison with control. The results with purified reuterin suggested that the development of slightly rosy colour in cheese was related to some compound produced/overproduced when higher levels of glycerol were present in cheese, but not due to reuterin. Application of L. reuteri INIA P572 as adjunct to the commercial starter with 100 mM glycerol led to such a reuterin concentration in cheese that could control undesirable microorganisms, avoiding the presence of color-changing compounds.

In vitro toxicity of reuterin, a potential food biopreservative.

Reuterin has a high potential as a food preservative due to both its chemical characteristics and its antimicrobial activity against food-borne pathogens and spoilage bacteria. However, there is a lack of information about its toxicity and its capacity to interfere with the metabolism of drugs by inhibiting cytochrome P450 (CYP) activity. The results of this study indicated that reuterin exhibited a moderate cytotoxicity in the human hepatoma cell line HepG2 according to assays measuring three different endpoints in the same set of cells. Reuterin was much less toxic than acrolein and only four times more toxic than diacetyl, a generally recognized as safe flavoring compound. In vitro experiments utilizing human liver microsomes showed that reuterin presents low possibility of displaying in vivo drug interactions by inhibition of CYP3A4, CYP2D6, and CYP2C9. Therefore, reuterin can be considered a promising food biopreservative, although additional toxicology research is needed before permission for use can be granted.

Natural antimicrobials and high-pressure treatments on the inactivation of Salmonella Enteritidis and Escherichia coli O157:H7 in cold-smoked salmon.

BACKGROUND: High hydrostatic pressure (HHP) combined with reuterin and lactoperoxidase system (LPS) has exerted antimicrobial activity against Listeria monocytogenes in cold-smoked salmon at chilled temperatures. Therefore the purpose of this work was to evaluate the effect of HHP combined with reuterin, LPS and lactoferrin (LF) on the survival of Salmonella enterica subsp. enterica serovar Enteritidis and Escherichia coli O157:H7 in cold-smoked salmon stored at 4 and 10 °C. RESULTS: Salmonella Enteritidis and E. coli O157:H7 were reduced more than 3 log colony-forming units (CFU) g(-1) by the pressure treatment (450 MPa/5 min). LPS slightly diminished pathogen levels throughout storage, whereas no effect was recorded when reuterin or LF was added. The Salmonella population was below the detection limit (<1 log CFU g(-1) ) during the storage of HHP-treated smoked salmon at 4 and 10 °C. The antimicrobial activity of HHP against E. coli O157:H7 was increased when 450 MPa was applied in combination with LPS in cold-smoked salmon at 4 and 10 °C. CONCLUSION: HHP at 450 MPa/5 min inactivated S. Enteritidis in cold-smoked salmon and in combination with LPS would be useful as a hurdle technology approach against E. coli O157:H7, even under mild temperature abuse conditions. © 2015 Society of Chemical Industry.