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    Peroxynitrous Acid Generated In Situ from Acidified H2O2 and NaNO2. A Suitable Novel Antimicrobial Agent?
    (Basel : MDPI, 2021) Balazinski, Martina; Schmidt-Bleker, Ansgar; Winter, Jörn; Woedtke, Thomas von
    Peroxynitrite (ONOO−) and peroxynitrous acid (ONOOH) are known as short acting reactive species with nitrating and oxidative properties, which are associated with their antimicrobial effect. However, to the best of our knowledge, ONOOH/ONOO- are not yet used as antimicro-bial actives in practical applications. The aim is to elucidate if ONOOH generated in situ from acidified hydrogen peroxide (H2O2 ) and sodium nitrite (NaNO2 ) may serve as an antimicrobial active in disinfectants. Therefore, the dose-response relationship and mutagenicity are investigated. Antimicrobial efficacy was investigated by suspension tests and mutagenicity by the Ames test. Tests were conducted with E. coli. For investigating the dose-response relationship, pH values and concentrations of H2O2 and NaNO2 were varied. The antimicrobial efficacy is correlated to the dose of ONOOH, which is determined by numerical computations. The relationship can be described by the efficacy parameter W, corresponding to the amount of educts consumed during exposure time. Sufficient inactivation was observed whenever W ≥ 1 mM, yielding a criterion for inactivation of E. coli by acidified H2O2 and NaNO2 . No mutagenicity of ONOOH was noticed. While further investigations are necessary, results indicate that safe and effective usage of ONOOH generated from acidified H2O2 and NaNO2 as a novel active in disinfectants is conceivable. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
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    Optimizing the application of plasma functionalised water (PFW) for microbial safety in fresh-cut endive processing
    (New York, NY [u.a.] : Elsevier Science, 2021) Schnabel, Uta; Balazinski, Martina; Wagner, Robert; Stachowiak, Jörg; Boehm, Daniela; Andrasch, Mathias; Bourke, Paula; Ehlbeck, Jörg
    The microbiological profiles and responses of native microflora of endive were investigated using a model process line, to establish where a defined PFW should be optimally applied to retain or improve produce microbiological quality. The PFW processes were compared with tap water and ClO2. The antimicrobial efficacy of PFW was quantified by determining the reduction in microbial load, the microbial viability and vitality. Depending on the stage of application of PFW, up to 5 log10-cycles reduction was achieved, accompanied by a reduction of metabolic activity, but not necessarily with a decrease in metabolic vitality. Multiple application (3-step-PFW-application) was more effective than single application (1-step-PFW-application) and PFW showed stronger antimicrobial effect in pre-cleaned endive. High concentrations of nitrite (315 mg l−1) and nitrate (472 mg l−1) in PFW were the main factors for the antimicrobial efficacy of PFW against bacteria. Furthermore, H2O2 and an acidic pH supported the mechanism of action against the endive microflora. These results identify the pathway to scale up successful industrial application of PFW targeting microbiological quality and safety of fresh leafy products.Industrial relevance The safety, quality and shelf life of freshly cut vegetables, e.g. lettuce, are strongly influenced by the microbial load. In addition, the hygienic design of production line, and a good handling/ production practice are indispensable. This study shows that the application of PFW, as a promising non-thermal sanitation technology, enables the inactivation of native microbial contamination on fresh-cut endive depending on the process stage of application. It further describes the impact of PFW on the metabolic activity and metabolic vitality of the lettuce-associated microflora. For higher acceptance, the mechanism of action of PFW was assumed based on previous chemical analyses and compared to the industrial standard of ClO2. The results contribute to the understanding and product-specificity of PFW-induced effects on safety, quality and shelf life of fresh cut lettuce and could be a basis for a possible industrial implementation and complement of common technologies.
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    Assessment of Phycocyanin Extraction from Cyanidium caldarium by Spark Discharges, Compared to Freeze-Thaw Cycles, Sonication, and Pulsed Electric Fields
    (Basel : MDPI, 2021) Sommer, Marie-Christine; Balazinski, Martina; Rataj, Raphael; Wenske, Sebastian; Kolb, Juergen F.; Zocher, Katja
    Phycocyanin is a blue colored pigment, synthesized by several species of cyanobacteria and red algae. Besides the application as a food-colorant, the pigmented protein is of high interest as a pharmaceutically and nutritionally valuable compound. Since cyanobacteria-derived phycocyanin is thermolabile, red algae that are adapted to high temperatures are an interesting source for phycocyanin extraction. Still, the extraction of high quality phycocyanin from red algae is challenging due to the strong and rigid cell wall. Since standard techniques show low yields, alternative methods are needed. Recently, spark discharges have been shown to gently disintegrate microalgae and thereby enable the efficient extraction of susceptible proteins. In this study, the applicability of spark discharges for phycocyanin extraction from the red alga Cyanidium caldarium was investigated. The efficiency of 30 min spark discharges was compared with standard treatment protocols, such as three times repeated freeze-thaw cycles, sonication, and pulsed electric fields. Input energy for all physical methods were kept constant at 11,880 J to ensure comparability. The obtained extracts were evaluated by photometric and fluorescent spectroscopy. Highest extraction yields were achieved with sonication (53 mg/g dry weight (dw)) and disintegration by spark discharges (4 mg/g dw) while neither freeze-thawing nor pulsed electric field disintegration proved effective. The protein analysis via LC-MS of the former two extracts revealed a comparable composition of phycobiliproteins. Despite the lower total concentration of phycocyanin after application of spark discharges, the purity in the raw extract was higher in comparison to the extract attained by sonication.