Soil protein as a potential antimicrobial agent against methicillin -resistant Staphylococcus aureus.

Recently, the interest is increasing to find alternatives to replace the usage of antibiotics since their massive and improper usage enhance the antibiotic resistance in human pathogens. In this study, for the first time we showed that the soil proteins have very high antibacterial activity (98% of growth inhibition) against methicillin resistant Staphylococcus aureus (MRSA), one of the most threatening human pathogens. We found that the protein extract (C3) from the forest with past intensive management showed higher antibacterial activity than that of unmanaged forest. The MIC and IC50 were found to be 30 and 15.0 µg protein g-1 dry soil respectively. C3 was found to kill the bacteria by cell wall disruption and genotoxicity which was confirmed by optical and fluorescent microscopy and comet assay. According to qPCR study, the mecA (the antibiotic resistant gene) expression in MRSA was found to be down-regulated after C3 treatment. In contrast, C3 showed no hemolytic toxicity on human red blood cells which was confirmed by hemolytic assay. According to ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS), 144 proteins were identified in C3 among which the majority belonged to Gram negative bacteria (45.8%). Altogether, our results will help to develop novel, cost-effective, non-toxic and highly efficient antibacterial medicines from natural sources against antibiotic resistant infections.

Induction of Laccase, Lignin Peroxidase and Manganese Peroxidase Activities in White-Rot Fungi Using Copper Complexes.

Ligninolytic enzymes, such as laccase, lignin peroxidase and manganese peroxidase, are biotechnologically-important enzymes. The ability of five white-rot fungal strains Daedaleopsis confragosa, Fomes fomentarius, Trametes gibbosa, Trametes suaveolens and Trametes versicolor to produce these enzymes has been studied. Three different copper(II) complexes have been prepared ((Him)[Cu(im)4(H2O)2](btc)·3H2O, where im = imidazole, H3btc = 1,3,5-benzenetricarboxylic acid, [Cu3(pmdien)3(btc)](ClO4)3·6H2O) and [Cu3(mdpta)3(btc)](ClO4)3·4H2O, where pmdien = N,N,N',N'',N''-pentamethyl-diethylenetriamine and mdpta = N,N-bis-(3-aminopropyl)methyl- amine), and their potential application for laccase and peroxidases induction have been tested. The enzyme-inducing activities of the complexes were compared with that of copper sulfate, and it has been found that all of the complexes are suitable for the induction of laccase and peroxidase activities in white-rot fungi; however, the newly-synthesized complex M1 showed the greatest potential for the induction. With respect to the different copper inducers, this parameter seems to be important for enzyme activity, which depends also on the fungal strains.

A comprehensive study of food waste management and processing in the Czech Republic: Potential health risks and consumer behavior.

Food waste is currently a widely discussed phenomenon with significant economic and social consequences. One third of the food produced in the world is wasted at various points along the food supply chain. This article presents a comprehensive study that examines consumer behavior in dealing with food waste and activities in the composting process that enable waste sanitation. The survey conducted as part of this study showed that consumers want to eliminate odors, are concerned about potential infections, and generally sort less food waste. This study suggested that the addition of appropriate additives could be a solution. The results indicated that additives could eliminate negative side effects such as unpleasant odors, the presence of insects and rodents, and act as a prevention of the occurrence of pathogenic organisms. Tea tree oil showed the best positive physical and chemical properties among the additives tested (CaCO3 and citric acid) with a significant effect on inhibiting the growth of bacterial strains such as Salmonella strains and had the strongest antibacterial effect, neutralized unpleasant odors, and stabilized the waste. The use of additives could be a future solution to meet consumer demands, improve the quality of food waste and advance the circular economy to improve the sustainability of agricultural systems.

Closing the loop: A case study on pathways for promoting sustainable waste management on university campuses.

The implementation of circular economy (CE) strategies has facilitated a comprehensive approach to waste management (WM) in university campuses. Composting food waste (FW) and biomass can mitigate negative environmental impacts and be part of a closed-loop economy. The compost can be used as a fertilizer, thereby closing the waste cycle. Implementing nudging strategies to promote effective waste segregation can help the campus move closer towards achieving neutrality and sustainability goals. The research was conducted at the Warsaw University of Life Sciences - WULS (SGGW). The University Campus is located in the south of Warsaw (Poland) and covers an area of 70 ha with 49 buildings. The SGGW campus generates selectively collected (glass, paper, plastic and metals, and biowaste) and mixed waste. Data were collected through a year-long report from the university administration. For the survey, waste data from 2019 to 2022 were obtained. The CE efficiency indicators of CE were measured. The indicators of CE efficiency for compost (Ic,ce) and plastic (Ipb,ce) showed Ic,ce at 21.05 %, which means that 1/5th of the waste generated on the campus can be introduced into the CE paradigm through composting, and the resulting value Ipb,ce of 19.96 % indicates that this amount can be reintroduced into the CE paradigm through its reuse. The results of the seasonality study showed that there were no statistically significant differences in the generated biowaste between the separated periods of the year, and the Pearson correlation coefficient (r = 0.068) provided additional confirmation. The weak correlation between the amount of biowaste generated and the average for each year (r = 0.110) also indicates a stable biowaste generation system that does not require a reduction or increase in the efficiency of waste processing, such as composting. By implementing CE strategies, university campuses can improve WM practices and achieve sustainability goals.

Direct microwave treatment enhances antioxidant and antibacterial properties of the seed extracts of Kékfrankos grapes.

The Kékfrankos is the most frequently cultivated wine grape in Hungary, with a significant national and regional impact, resulting in considerable amounts of byproducts (e.g. pomace, seeds). To the best of our knowledge no research has been conducted on the antioxidant and antibacterial properties of its seed extracts (GSE). A novel apporach of applying direct microwave treatment on grape seeds was implemented for the first time to enhance antioxidant and antimicrobial properties of GSE. Antioxidant properties were assayed using the DPPH (2,2-diphenyl-1-picrylhydrazyl), FRAP (Ferric Reducing Antioxidant Power) and TPC (Folin-Ciocâlteu's Total Polyphenol Content) methods. Profile and content of polyphenols was studied using high-performance liquid chromatography/tandem mass spectrometry and matrix-assisted laser desorption/ionization mass spectrometry. Antibacterial properties were evaluated using Gram-positive Staphylococcus aureus (SA), methicillin-resistant Staphylococcus aureus (ST239) (MRSA) and Gram-negative Escherichia coli (EC) bacteria strains. Results proved that the mild direct microwave treatment of grape seeds significantly increased total polyphenol, (+)-catechin, (-)-epicatechin as well as antioxidant capacity levels by 20-30 % compared to untreated samples and resulted the best antibacterial properties based on bacterial growth curves (SA and MRSA: 0.015625 mg/mL, EC: 0.25 mg/mL). Results justify the importance of further pharmacological investigations on Kékfrankos grape seed extracts and that the direct microwave treatment of grape seeds is an innovative approach for the fast and cost efficient improvement of the antibacterial properties of grape seed extracts.

Food waste composting - Is it really so simple as stated in scientific literature? - A case study.

Food waste has recently gained much worldwide interest due to its influence on the environment, economy and society. Gathering and recycling of food waste is the essential issue in the waste management and the interest in processing food waste arises mainly out of influence of the processes of food putrefaction on the environment. Composting of food waste encounters a number of technical challenges, arising weak physical structure of food waste with weak porosity, high content of water, low carbon-to-nitrogen relation and fast hydrolysis and accumulation of organic acids during composting. Therefore, the aim of this study was to investigate the challenges facing installations intended for food waste composting, with the purpose to their optimization with use of appropriate additives. Physico-chemical, biochemical characteristics and phytotoxicity of the produced compost has been measured. Two additives (20% biochar and 20% sawdust) were chosen from experimental variants I-XII containing different additives (biochar, Devonian sand, sawdust) in diverse concentration. The use of selected additives seems to slightly increase potential of hydrogen value and carbon-to-nitrogen ratio, while decreasing electrical conductivity in comparison with control sample. The results obtained also show that the addition of biochar leads to an increase dehydrogenase, phosphatase and arylsulphatase activities and addition of sawdust has a positive effect on beta-D-glucosidase, protease, phosphatase and arylsulphatase activities. The phytotoxicity test shows that the compost made of food waste (control sample) and with addition of biochar is toxic to plants. By contrast, the addition of sawdust shows that the compost was not phytotoxic. In conclusion, the addition of additives does not provide unambiguous results in terms of the quality of the final product in all monitored parameters. Therefore, we can state that food waste was reduced and hygienized, and that the final product does not meet conditions for mature compost.

Composting versus mechanical-biological treatment: Does it really make a difference in the final product parameters and maturity.

One of crucial waste management problems is the management of organic waste. This activity employs the composting. In case of green waste, its application seems reasonable, whereas the use of selected mixed waste raises problems related to the compost quality. Across countries, the non-sterile organic fraction of municipal solid waste is being separated through the mechanical-biological treatment. The technology is a solution of waste treatment and meets objectives set out in the Landfill Directive. There are many problems associated with the use of output products. The use of compost as a fertilizer requires determination of its impact on the environment. Compost quality can be assessed using analytical methods and phytotoxicity tests. Therefore, the aim of this study was to describe changes in physico-chemical, enzymatic, phytotoxicity and vegetation parameters occurring in composts from two systems - a prismatic installation for green waste, and a mechanical-biological treatment installation. The compost from green waste exhibited greater stability. Values of dehydrogenase activity were lower if compared with the mechanically and biologically treated compost, which indicates lower compost maturity. The biomass production of Brassica napus L. and Fetuca rubra L. was higher in the variant with the application of green compost. The influence on Hordeum vulgare L., Cannabis sativa L., and Sinapis alba L. depended on the plant type and the compost used. Nevertheless, the compost from green waste was less toxic. The evidence from this study suggests that the mechanical-biological treatment had problems associated with the maturation and quality of the final product.

Use of soil enzyme activities to assess the recovery of soil functions in abandoned coppice forest systems.

Coppicing consists of periodically cutting back tree stems to ground level to stimulate the growth of multiple stems from the stool. In Central Europe, many coppiced forests were abandoned at the beginning of the last century owing to a decline in the demand for charcoal and wood. This was assumed to enable the forests to recover and the properties to become similar to those of unmanaged forest (high forest). Most studies on abandoned coppiced forest have focused on forest recovery, while soil recovery has generally been overlooked. With the aim of filling this gap, this study investigated the effect of coppicing abandonment on soil recovery by analysing the changes in soil enzyme activities (dehydrogenase, ß-glucosidase, invertase, urease, acid phosphatase and arylsulphatase). Two differently managed sessile oak (Quercus petraea) forests were selected for study: a former coppice forest, abandoned >90 years ago, and an undisturbed forest. The analytical data were compared to assess the degree of recovery of the soil in the abandoned coppice forest. The soil organic matter content was two times lower in the abandoned coppice than in the high forest, suggesting that organic matter depletion due the past coppicing is a long-term effect. All of the absolute enzyme activities were also two times lower in the abandoned coppice forest soil than in the high forest soil. However, the specific enzyme activities were similar in both types of soil. This indicates that metabolic activity is similar in both soil types, suggesting that it either recovers faster than organic matter and soil enzyme activity or that, despite the depletion in organic matter and enzyme activities, metabolic activity was sustained in coppiced forest soil. However, in the latter case this would imply that organic matter and soil enzymes were lost in exactly the same proportion, which is highly improbable.

Immobilization of ligninolytic enzymes from white-rot fungi in cross-linked aggregates.

Ligninolytic enzymes from white-rot fungi are widely used in biotechnological processes. However, the application of these enzymes as free enzymes is limited due to their instability and lack of reusability. Enzyme stabilization is therefore a major challenge in biocatalytic process research, and immobilization methods are desirable. Using cross-linked enzyme aggregates (CLEAs) such as magnetic CLEAs, porous-CLEAs and combi-CLEAs is a promising technique for overcoming these issues. Cross-linking methods can stabilize and immobilize enzymes by interconnecting enzyme molecules via multiple bonds using cross-linking agents such as glutaraldehyde. The high catalyst density and microporous assembly of CLEAs guarantee high catalyst activity, which, together with their long shelf life, operational stability, and reusability, provide a cost-efficient alternative to matrix-assisted immobilization approaches. Here, we review current progress in ligninolytic enzyme immobilization and provide a comprehensive review of CLEAs. Moreover, we summarize the use of these CLEAs for biocatalysis processes, bioremediation such as dye decolourization, wastewater treatment or pharmaceutically active compound elimination.

Preparation and Optimisation of Cross-Linked Enzyme Aggregates Using Native Isolate White Rot Fungi Trametes versicolor and Fomes fomentarius for the Decolourisation of Synthetic Dyes.

The key to obtaining an optimum performance of an enzyme is often a question of devising a suitable enzyme and optimisation of conditions for its immobilization. In this study, laccases from the native isolates of white rot fungi Fomes fomentarius and/or Trametes versicolor, obtained from Czech forests, were used. From these, cross-linked enzyme aggregates (CLEA) were prepared and characterised when the experimental conditions were optimized. Based on the optimization steps, saturated ammonium sulphate solution (75 wt.%) was used as the precipitating agent, and different concentrations of glutaraldehyde as a cross-linking agent were investigated. CLEA aggregates formed under the optimal conditions showed higher catalytic efficiency and stabilities (thermal, pH, and storage, against denaturation) as well as high reusability compared to free laccase for both fungal strains. The best concentration of glutaraldehyde seemed to be 50 mM and higher efficiency of cross-linking was observed at a low temperature 4 °C. An insignificant increase in optimum pH for CLEA laccases with respect to free laccases for both fungi was observed. The results show that the optimum temperature for both free laccase and CLEA laccase was 35 °C for T. versicolor and 30 °C for F. fomentarius. The CLEAs retained 80% of their initial activity for Trametes and 74% for Fomes after 70 days of cultivation. Prepared cross-linked enzyme aggregates were also investigated for their decolourisation activity on malachite green, bromothymol blue, and methyl red dyes. Immobilised CLEA laccase from Trametes versicolor showed 95% decolourisation potential and CLEA from Fomes fomentarius demonstrated 90% decolourisation efficiency within 10 h for all dyes used. These results suggest that these CLEAs have promising potential in dye decolourisation.

Effect of inoculation with white-rot fungi and fungal consortium on the composting efficiency of municipal solid waste.

An investigation was carried out on the effect of inoculation methods on the compost of an organic fraction of municipal solid waste. Three types of white-rot fungi (Phanerochaete chrysosporium, Trametes versicolor and Fomes fomentarius), and a consortium of these fungi, were used. The study assessed their influence on microbial enzymatic activities and the quality of the finished compost. It was found that the addition of white-rot fungi to municipal solid waste (after 37days of composting) could be a useful strategy for enhancing the properties of the final compost product. In comparison with the control sample (compost without inoculation), it accelerates degradation of solid waste as indicated by changes in C/N, electrical conductivity and pH. However, the effectiveness of waste degradation and compost maturation depends on the type of microorganism used for inoculation. The presence of inoculants, such as Trametes versicolor and Fomes fomentarius, led to a higher degrading ratio and a better degree of maturity. This resulted in an increase of enzymatic activities (especially dehydrogenase and protease) and a germination index in comparison with inoculation using Phanerochaete chrysosporium or a consortium of fungi.

Biofilm formation and extracellular polymeric substances (EPS) production by Bacillus subtilis depending on nutritional conditions in the presence of polyester film.

The influence of biofilm formation as the mode of microorganism growth on degradation of synthetic polymers represents an important research topic. This study focuses on the effect of biofilm developed by Bacillus subtilis (BS) cultivated submerged under various nutrition conditions on biodeterioration of poly(ε-caprolactone) film. Polymer in the film form (thickness 0.7 mm) was incubated for 21 days either continuously or by regularly renewed system. The scission of polyester chain bonds took place in all biotic media and was enhanced by biofilm formation in nutrient-rich media.