Preliminary Study on the Effect of Fermented Cheese Whey on Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella Goldcoast Populations Inoculated onto Fresh Organic Lettuce.

Cheese whey fermented by an industrial starter consortium of lactic acid bacteria was evaluated for its antibacterial capacity to control a selection of pathogenic bacteria. For their relevance on outbreak reports related to vegetable consumption, this selection included Listeria monocytogenes, serotype 4b, Escherichia coli O157:H7, and Salmonella Goldcoast. Organically grown lettuce was inoculated with an inoculum level of ∼10(7) colony-forming unit (CFU)/mL and was left for about 1 h in a safety cabinet before washing with a perceptual solution of 75:25 (v/v) fermented whey in water, for 1 and 10 min. Cells of pathogens recovered were then counted and their number compared with that obtained for a similar treatment, but using a chlorine solution at 110 ppm. Results show that both treatments, either with chlorine or fermented whey, were able to significantly reduce (p < 0.05) the number of bacteria, in a range of 1.15-2.00 and 1.59-2.34 CFU/g, respectively, regarding the bacteria tested. Results suggest that the use of fermented whey may be as effective as the solution of chlorine used in industrial processes in reducing the pathogens under study (best efficacy shown for Salmonella), with the advantage of avoiding health risks arising from the formation of carcinogenic toxic chlorine derivates.

Bridging the Gap to Non-toxic Fungal Control: Lupinus-Derived Blad-Containing Oligomer as a Novel Candidate to Combat Human Pathogenic Fungi.

The lack of antifungal drugs with novel modes of action reaching the clinic is a serious concern. Recently a novel antifungal protein referred to as Blad-containing oligomer (BCO) has received regulatory approval as an agricultural antifungal agent. Interestingly its spectrum of antifungal activity includes human pathogens such as Candida albicans, however, its mode of action has yet to be elucidated. Here we demonstrate that BCO exerts its antifungal activity through inhibition of metal ion homeostasis which results in apoptotic cell death in C. albicans. HIP HOP profiling in Saccharomyces cerevisiae using a panel of signature strains that are characteristic for common modes of action identified hypersensitivity in yeast lacking the iron-dependent transcription factor Aft1 suggesting restricted iron uptake as a mode of action. Furthermore, global transcriptome profiling in C. albicans also identified disruption of metal ion homeostasis as a potential mode of action. Experiments were carried out to assess the effect of divalent metal ions on the antifungal activity of BCO revealing that BCO activity is antagonized by metal ions such as Mn2+, Zn2+, and Fe2+. The transcriptome profile also implicated sterol synthesis as a possible secondary mode of action which was subsequently confirmed in sterol synthesis assays in C. albicans. Animal models for toxicity showed that BCO is generally well tolerated and presents a promising safety profile as a topical applied agent. Given its potent broad spectrum antifungal activity and novel multitarget mode of action, we propose BCO as a promising new antifungal agent for the topical treatment of fungal infections.

Blad-Containing Oligomer Fungicidal Activity on Human Pathogenic Yeasts. From the Outside to the Inside of the Target Cell.

Blad polypeptide comprises residues 109-281 of Lupinus albus ß-conglutin precursor. It occurs naturally as a major subunit of an edible, 210 kDa oligomer which accumulates to high levels, exclusively in the cotyledons of Lupinus seedlings between the 4th and 14th day after the onset of germination. Blad-containing oligomer (BCO) exhibits a potent and broad spectrum fungicide activity toward plant pathogens and is now on sale in the US under the tradename FractureTM. In this work we demonstrate its antifungal activity toward human pathogens and provide some insights on its mode of action. BCO bioactivity was evaluated in eight yeast species and compared to that of amphotericin B (AMB). BCO behaved similarly to AMB in what concerns both cellular inhibition and cellular death. As a lectin, BCO binds strongly to chitin. In addition, BCO is known to possess 'exochitinase' and 'endochitosanase' activities. However, no clear disruption was visualized at the cell wall after exposure to a lethal BCO concentration, except in cell buds. Immunofluorescent and immunogold labeling clearly indicate that BCO enters the cell, and membrane destabilization was also demonstrated. The absence of haemolytic activity, its biological origin, and its extraordinary antifungal activity are the major outcomes of this work, and provide a solid background for a future application as a new antifungal therapeutic drug. Furthermore, its predictable multisite mode of action suggests a low risk of inducing resistance mechanisms, which are now a major problem with other currently available antifungal drugs.