Antimicrobial properties of lyophilized extracts of olive fruit, pomegranate and orange peel extracts against foodborne pathogenic and spoilage bacteria and fungi in vitro and in food matrices

Several novel antimicrobials with different concentrations of olive, pomegranate, and orange fruit pulp extracts were produced from agricultural byproducts and, after lyophilization, their antimicrobial activity and potential synergistic effects were evaluated in vitro and in food samples against foodborne pathogenic and spoilage bacteria and fungi. The Minimum Inhibitory of the tested bacteria was 7.5% or 10%, while fungi were inhibited at a concentration of 10% or above. The optical density of bacterial and yeast cultures was reduced to a different extent with all tested anti-microbial powders, compared to a control without antimicrobials, and mycelium growth of fungi was also restricted with extracts containing at least 90% olive extract. In food samples with inoculated pathogens and spoilage bacteria and fungi, the 100% olive extract was most inhibitory against E. coli, S. typhimurium, and L. monocytogenes in fresh burger and cheese spread samples (by 0.6 to 1.8 log cfu/g), except that S. typhimurium was better inhibited by a 90% olive and 10% pomegranate extract in burgers. The latter extract was also the most effective in controlling the growth of inoculated fungi (Αspergillus niger, Penicillium italicum, Rhodotorula mucilaginosa) in both yogurt and tomato juice samples, where it reduced fungal growth by 1–2.2 log cfu/g at the end of storage period. The results demonstrate that these novel encapsulated extracts could serve as natural antimicrobials of wide spectrum, in order to replace synthetic preservatives in foods and cosmetics. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

The effect of agitation and aeration on the synthesis and molecular weight of gellan in batch cultures of sphingomonas paucimobilis

The effects of agitation and aeration upon synthesis and molecular weight of the biopolymer gellan were systematically investigated in batch fermenter cultures of the bacterium, Sphingomonas paucimobilis. High aeration rates and vigorous agitation enhanced growth of S. paucimobilis. Although gellan formation occurred mainly in parallel with cell growth, the increase in cells able to synthesise gellan did not always lead to high gellan production. For example, at very high agitation rates (1000 rpm) growth was stimulated at the expense of biopolymer synthesis. Maximal gellan concentration was obtained at 500 rpm agitation and either 1 or 2 vvm aeration (12.3 and 12.4 g/l gellan, respectively). An increase in aeration (from 1 to 2 vvm) enhanced gellan synthesis only at low agitation rates (250 rpm). However, high aeration or dissolved oxygen was not necessary for high gellan synthesis, in fact oxygen limitation always preceded the phase of maximum gellan production and probably enhanced polysaccharide biosynthesis. Some gellan was formed even after glucose exhaustion. This was attributed to the intracellular accumulation of polyhydroxyalkanoates, (such as polyxydroxybutyrate) which were found in S. paucimobilis cells indicating the existence of a carbon storage system, which may contribute to gellan biosynthesis under glucose-limiting conditions. The autolysis of the culture, which occurred at the late stages of the process, seemed to be triggered mainly by limitations in mass (nutrient) transfer, due to the highly viscous process fluid that gradually develops. Rheological measurements generally gave a very good near real time estimate of maximum biopolymer concentration offering the possibility of improved process control relative to time consuming gravimetric assay methods. While mechanical depolymerisation of gellan did not occur, high aeration rates (2 vvm) led to production of gellan of low molecular weight (at either 250 or 500 rpm). This effect of aeration rate upon gellan molecular weight is reported here for the first time, and is important for the properties and applications of gellan. Mechanisms which may have led to this are discussed, but control of molecular weight of the biopolymers is clearly an area needing further research

Vacuum microwave-assisted aqueous extraction of polyphenolic compounds from avocado (Persea americana) solid waste

The extraction efficacy of avocado fruit peels (AP) and seeds (AS) with the use of vacuum microwave-assisted aqueous extraction (VMAAE) was optimized in this study, with regard to extract’s total phenolic content (TPC), maximum antioxidant activity and minimal operational cost. Temperature (79.64 and 78.11 °C for AP and 43.90 and 45.26 °C for AS), time (11.89 and 11.75 min for AP, 10.18 and 10.28 min for AS), ratio of water to raw material (16.45% and 10.02% for AP, 38.73% and 37.65% for AS) and microwave power (5708.04 and 5699.10 W for AP, 5549.08 and 4797.29 W for AS) were estimated statistically as the optimal conditions in order to achieve high rates of extracts with high TPC and antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH●) scavenging radical methods, respectively. VMAAE performed under these conditions resulted in received extracts with TPC (0.352 gallic acid equivalent-GAE/g fresh AP/min and 0.124 GAE/g fresh AS/min). Furthermore, it was calculated the DPPH● radical scavenging activity was equal to 100 mg/L expressed in L of 0.104 L/min for AP and 0.045 L/min for AS. The results of our study may give a promising solution to avocado processing companies for further utilization of their waste. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Optimization of vacuum microwave-assisted extraction of pomegranate fruits peels by the evaluation of extracts’ phenolic content and antioxidant activity

The global interest in the use of plant by-product extracts as functional ingredients is continuously rising due to environmental, financial and health benefits. The latest advances in extraction technology have led to the production of aqueous extracts with high bioactive properties, which do not require the use of organic solvents. The purpose of this study was to optimize the conditions applied for the extraction of pomegranate peels (PP) via a “green” industrial type of vacuum microwave-assisted aqueous extraction (VMAAE), by assessing the potential bioactivity of the extracts (in terms of phenolic content and antioxidant activity), using a response surface methodology. The extraction conditions of temperature, microwave power, time and water/PP ratio were determined by the response surface methodology, in order to yield extracts with optimal total phenolics concentrations (TPC) and high antioxidant activity, based on the IC50 value of the scavenging of the 2,2-diphenyl-1-picrylhydrazyl (DPPH•) radical. The values of the optimum extraction parameters, such as extraction temperature (61.48 and 79.158◦C), time (10 and 12.17 min), microwave power (3797.24 and 3576.47 W) and ratio of water to raw material (39.92% and 38.2%), were estimated statistically for the two responses (TPC and IC50 values), respectively. Under these optimal extraction conditions, PP extracts with high TPC ((5.542 mg Gallic Acid Equivalent (GAE)/g fresh PP))/min and radical scavenging activity (100 mg/L (1.6 L/min)) could be obtained. Our results highlighted that the optimized industrial type of VMAAE could be a promising solution for the valorization of the PP by-products. © 2020 by the authors. Licensee MDPI, Basel, Switzerland

Microbiomic Prospects in Fermented Food and Beverage Technology

Fermentation allows production of healthy, preservable, and nutritious foodstuff and beverages, such as respective dairy and meat products, vegetables (pickles) and olives, beer, and wine, where a wide array of microbiota, both prokaryotes and eukaryotes, participate simultaneously or successively, creating distinct though multiple growth and developmental dynamics in terms of succession and prevalence of different genders of cellular microbiota, during and after fermentation. Genomic methodology reveals and elucidates the diversity of similar populations in different localities, enabling a more qualitative marketing of products. Simultaneously, spoilage and pathogenic microbial taxa contaminate and degrade fermented foodstuff, adversely affecting quality, shelf-life, and safety of the final product, which can be stratified by genomic and metabolomic approaches to further detail than hitherto. The positive or negative interactions of productive and counterproductive taxa within these microbiomes and their cumulative effect define the success of the process in terms of marketing efficiency, thus establishing practices of using either indigenous microbiota or added starter cultures, each with different effects on quality, sensory characteristics, storability, and safety. © 2020 Elsevier Inc. All rights reserved

Effect of Different Formulations of Polyphenolic Compounds Obtained from OMWW on the Growth of Several Fungal Plant and Food Borne Pathogens. Studies in vitro and in vivo

AbstractThe assessment of different concentrations and forms of polyphenols (free and encapsulated) from olive mill waste (OMWW) as plant protection materials against economically important plant pathogenic fungi in in vitro and in vivo tests is the main research area of this work. In the first stage of the experimental process, it was assessed the zone of inhibition in mm of several fungal pathogens using the methods of disk diffusion assay, and well diffusion assay. In the second stage of the evaluation, the effect of polyphenols against 14 fungal microorganisms was examined in order to determine the MIC / microbicidal concentration (MIC / MFC). In a later stage in vivo evaluation of liquid polyphenols (LFP) obtained from OMWW as natural bio-chemicals against several fungal pathogens on tomato plants was carried. The evaluation of the results obtained by determining the MIC and MLC, demonstrated that the fungus Aspergillus flavus appeared highly resistant to the LFP concentration required minimum sample rate > 35-40% for inhibition and killing effects respectively. In order of major protection resulting from the use of polyphenolic compound against major diseases, Botrytis cinerea, Sclerotinia sclerotiorum and Ascochyta lentis gave the most promising results. Moreover, the use of low concentration of LFP at 5 and 10% could control in some cases fungal pathogens. However, higher concentration of LFP (20 and 30%) appeared possible phytotoxic effects

Potential synergistic action of liquid olive fruit polyphenol extract with aqueous extracts of solid wastes of pomegranate or/and orange juice industry as organic phyto-protective agents against important plant pathogens - Part 1 (in vitro studies)

In this research study potential synergistic action of aqueous extracts of olive-pomegranate-orange fruits isolated by microwave assisted extraction were investigated as biocontrol agents against several plant pathogenic fungi. The aim of the research study was the production of mixed liquid extracts to be used as natural phytoprotective agents in organic farming. The results proved that the use of 100%-pure olive fruit polyphenol extract affected the mycelium growth of B. cinerea, V. dahliae, R. solani, E. lata, M. laxa, P. lycopersici and P. punicae. However, it did not affect the mycelium growth of F. oxysporum f. sp., lycopersici, A. niger, and G. graminis. Moreover, pomegranate polyphenol extract at 100% concentration inhibited mycelium growth of B. cinerea. Various olive-pomegranate-orange polyphenol aqueous extract combinations may be used in order to control the mycelium growth of the majority of the tested microorganisms. However, the use of any combination and concentrations of the tested samples (1-72) did not affect the mycelium growth of F. oxysporum f. sp., lycopersici, A. niger, and G. graminis thus, their use as biological control agent is not recommended. According to the findings of the present research, it can be suggested that the olive fruit polyphenol extract be used as potential biocontrol agent, in order to stimulate mycelium growth of various plant pathogenic fungi. Copyright © 2020 by authors, all rights reserved. Authors agree that this article remains permanently open access under the terms of the Creative Commons Attribution License 4.0 International Licens

Preliminary Studies on Suppression of Important Plant Pathogens by Using Pomegranate and Avocado Residual Peel and Seed Extracts

Potential synergistic action of aqueous extracts of pomegranate peel (PP), avocado peel (AP), and avocado seed (AS) wastes isolated by microwave-assisted extraction were assessed in in vitro and in vivo assays as biocontrol agents against several plant pathogenic fungi. The study findings contribute to the utilization of a value-added industrial byproduct and provide significant value in advancing the development of new plant protecting compositions that benefit from the synergistic effects between two important plant species that contain several natural bioactive compounds. More specifically, the in vitro results proved that the use of 100%-pure (PP) extracted waste affected the mycelium growth of Penicillium expansum. Furthermore, mycelium growth of Aspergillus niger was decreased by 10.21% compared to control after 7 days of growth in medium agar containing 100% AP and extracted waste. Moreover, mycelium growth of Botrytis cinerea was affected by equal volume of avocado extraction wastes (50% peel and 50% seed) only at the first 3 days of the inoculation, while at the seventh day of the inoculation there was no effect on the mycelium growth. Equal volumes of the examined wastes showed decreased mycelium growth of Fusarium oxysporum f.sp. lycopersici by 6%, while Rhizoctonia solani mycelium growth was found to be the most sensitive in PP application. In addition, the in vivo assay shown that PP extract suppresses damage of tomato plants caused by R. solani followed by extracted wastes from AP. Based on the research findings, it can be argued that PP and AP extracts can be used as natural antifungals instead of dangerous synthetic antifungals to effectively treat phytopathogens that cause fruit and vegetable losses during cultivation. © 2022 by the authors. Licensee MDPI, Basel, Switzerland