Safety assessment of the biotechnologically produced human-identical milk oligosaccharide 3-Fucosyllactose (3-FL).

3-Fucosyllactose (3-FL), a highly abundant complex carbohydrate in human breast milk, functions as a prebiotic promoting early microbial colonization of the gut, increasing pathogen resistance and modulating immune responses. To investigate potential health benefits, 3-FL was produced by fermentation using a genetically modified E. coli K12 strain. The safety assessment of 3-FL included acute oral toxicity, in vitro and in vivo assessment of genetic toxicity, and a subchronic rodent feeding study. 3-FL was not acutely toxic at 5000 mg/kg bw, and there was no evidence of genetic toxicity in the bacterial reverse mutation test and chromosomal aberration assay. There was a repeatable statistically-significant trend in the 4-h S9-activated test conditions in the in vitro micronucleus assay; the confirmatory in vivo mouse micronucleus study was negative at all doses. Dietary subchronic exposure of rats to 3-FL (5% and 10%) did not produce any statistical or biologically-relevant differences in growth, food intake or efficiency, clinical observations, or clinical or anatomic pathology changes at average daily intakes of 5.98 and 7.27 g/kg bw/day for males and females, respectively. The weight of evidence from these studies support the safe use of 3-FL produced using biotechnology as a nutritional ingredient in foods.

Inventory of microbial species with a rationale: a comparison of the IDF/EFFCA inventory of microbial food cultures with the EFSA Biohazard Panel qualified presumption of safety.

In order to provide a harmonised preassessment to support risk assessment performed by the European Food Safety Authority (EFSA), the Biohazard Panel in 2007 published guidelines for evaluation of the safety of a strain included in the food chain, the Qualified Presumption of Safety (QPS). Since 2008, the Biohazard Panel has published on a regular basis an update of the microbial strains submitted for approval and extends the list of species which have been granted QPS status. The International Dairy Federation (IDF) and the European Food and Feed Cultures Association (EFFCA) have, since 2002, been conducting a project on the safety demonstration of microbial food cultures (MFCs). Following the publication of IDF Bulletin 377-2002, an inventory of MFCs was published in IDF Bulletin 455-2012 and updated most recently in IDF Bulletin 495-2018. These two lists developed by EFSA (QPS) and IDF/EFFCA both propose as an outcome an inventory of microbial species that are safe for human consumption. To avoid confusion when these two inventories are compared, this review attempts to explain the rationale that was used to develop them and explain how the two lists should be understood.

Safety evaluation of Evesse EPC, an apple polyphenol extract rich in flavan-3-ols.

The safety of the apple polyphenol extract EvesseEPC, which is rich in flavan-3-ols, particularly epicatechin, was evaluated. Both in a bacterial reverse mutation test and a mouse lymphoma assay, EvesseEPC showed a positive response in vitro. In vivo studies (UDS test in hepatocytes, bone marrow micronucleus test and comet assay in intestinal cells) were all negative and hence Evesse EPC is considered not to have genotoxic properties in vivo. In a 90-day study in rats, EvesseEPC was administered at dietary levels of 0%, 1.25%, 2% and 3.25%. Body weights were decreased in the high-dose group in both sexes without effects on feed or water intake. In the high-dose group, thrombocytes (males) and creatinine (both sexes) were decreased, prothrombin time (males) was increased, and liver, kidneys and spleen weights were increased (males), without histological correlates. Diffuse acinar cell hypertrophy, observed in the parotid salivary glands in all treatment groups, was not considered as adverse and presumably reflected a local, reversible and adaptive response to direct contact with EvesseEPC. The NOAEL for EvesseEPC in rats was 2% in the diet, equivalent to an overall average intake of 1.3 and 1.5 g/kg body weight/day for males and females, respectively.

28-Day oral toxicity study in rats with high purity barley beta-glucan (Glucagel).

Beta-glucans are glucose polymers present in cereal grains, particularly barley and oat. Consumption of these grains or concentrated beta-glucan preparations has been shown to lower blood cholesterol. The present study was conducted to assess the safety of a high purity (>75%) barley beta-glucan (Glucagel). The product was fed to Wistar rats (5/sex/group) at dietary levels of 0% (control), 1%, 5% and 10% for 28 days. Clinical and neurobehavioural observations, growth, feed and water consumption, ophthalmoscopy, haematology, clinical chemistry, urinalysis, organ weights, necropsy and histopathological examination revealed no adverse effects of Glucagel. High-dose males exhibited lower plasma cholesterol and phospholipids levels and a higher plasma urea level. These slight changes were considered of no toxicological significance. Full and empty caecum weights were increased in mid- and high-dose males. This caecal enlargement was a physiological response to the consumption of a high amount of indigestible carbohydrate and considered of no toxicological concern. In conclusion, feeding Glucagel at dietary levels up to 10% for 28 days was tolerated without any signs of toxicity. This dietary level was equivalent to 7.7 g Glucagel (5.8 g beta-glucan)/kg body weight/day in male rats and 7.8 g Glucagel (5.9 g beta-glucan)/kg body weight/day in female rats.

Cloning and transfer of genes for antifungal compounds from Erwinia herbicola to Escherichia coli.

Erwinia herbicola CHS1065 produces antifungal compounds highly related to herbicolin A. In 11 Tn5 mutants that have lost the antifungal activity, the transposon insertion is located on a 170-kb plasmid present in CHS1065. This plasmid was designated pHER1065. When analyzing these antifungal mutants, it was found that the genes for the biosynthesis of the antifungal compounds were organized in at least two clusters on pHER1065. Upon insertion of the aphII gene of Tn5 and genes for plasmid mobilization in pHER1065, the plasmid could be stably introduced into Escherichia coli. All the E. coli exconjugants expressed an antifungal activity that was quantitatively and qualitatively comparable to the activity produced by E. herbicola CHS1065. Amino acid analysis and molecular weight determinations of the antifungal compound produced by CHS1065 were identical to those of herbicolin A.

[Genetic recombination and Fib genes transfer of the plasmid of E. herbicola CSH 1065].

We inserted kanamycin resistance (Km) gene and mobilized function (Mob) gene on the plasmid of E. herbicola CSH1065 by using DNA molecular cloning and genetic recombination techniques. Therefore, the plasmid of E. herbicola CSH1065 to E. coli HB101 could be transferred by conjugation. The expression of those genes concerning fungi inhibition function (Fib) of E. herbicola CSH1065 in E. coli HB101 was observed. This result confirmed again the fungi inhibition genes of E. herbicola CSH1065 is only related to its plasmid genome not to its chromosome genome. Meanwhile those yellow pigment genes located on the plasmid don't involve the antifungi function of E. herbicola CSH1065. All those results were convinced by DNA molecular hybridizations.

Transformation of Brassica napus and Brassica oleracea Using Agrobacterium tumefaciens and the Expression of the bar and neo Genes in the Transgenic Plants.

An efficient and largely genotype-independent transformation method for Brassica napus and Brassica oleracea was established based on neo or bar as selectable marker genes. Hypocotyl explants of Brassica napus and Brassica oleracea cultivars were infected with Agrobacterium strains containing chimeric neo and bar genes. The use of AgNO(3) was a prerequisite for efficient shoot regeneration under selective conditions. Vitrification was avoided by decreasing the water potential of the medium, by decreasing the relative humidity in the tissue culture vessel, and by lowering the cytokinin concentration. In this way, rooted transformed shoots were obtained with a 30% efficiency in 9 to 12 weeks. Southern blottings and genetic analysis of S1-progeny showed that the transformants contained on average between one and three copies of the chimeric genes. A wide range of expression levels of the chimeric genes was observed among independent transformants. Up to 25% of the transformants showed no detectable phosphinotricin acetyltransferase or neomycin phosphotransferase II enzyme activities although Southern blottings demonstrated that these plants were indeed transformed.