Four-decades evolutionary development of municipal solid waste management in China: Implications for sustainable waste management and circular economy.

This study collected data on waste generation and management in China between 1979 and 2020 from government statistics and literature and reviewed the development of municipal solid waste (MSW) management in China. The extended stochastic impact by regression on population, affluence and technology (STIRPAT) model was employed to identify the driving forces of MSW generation, and the cointegration analysis showed that economy (0.35, t = -3.47), industrial structure (3.34, t = -20.77) and urbanization (-1.5, t = 5.678) were the significant socioeconomic driving forces in the long run. By employing the framework of evolutionary economics, this study then investigated the internal rules of long-term interaction between socioeconomic factors and MSW management. The results indicate that, in the long run, MSW management development can be viewed as an evolutionary process that includes a continuous adaptation to external socioeconomic factors and the co-evolution of internal institutions and technologies. Adaptation and diversity of institutions and technologies play an important role in achieving sustainable waste management and circular economy (CE). This study offers a novel evolutionary perspective for explaining dynamic changes of MSW management in China, as well as recommendations for emerging economies to achieve sustainable waste management and CE goals.

Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications.

α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60-80 °C and is active and stable over a wide pH range (4.0-9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.

Antitumor activity of natural pigment violacein against osteosarcoma and rhabdomyosarcoma cell lines.

PURPOSE: Sarcomas are rare and heterogenic tumors with unclear etiology. They develop in bone and connective tissue, mainly in pediatric patients. To increase efficacy of current therapeutic options, natural products showing selective toxicity to tumor cells are extensively investigated. Here, we evaluated antitumor activity of bacterial pigment violacein in osteosarcoma (OS) and rhabdomyosarcoma (RMS) cell lines. METHODS: The toxicity of violacein was assessed in vitro and in vivo, using MTT assay and FET test. The effect of violacein on cell migration was monitored by wound healing assay, cell death by flow cytometry, uptake of violacein by fluorescence microscopy, generation of reactive oxygen species (ROS) by DCFH-DA assay and lipid peroxidation by TBARS assay. RESULTS: Violacein IC50 values for OS and RMS cells were in a range from 0.35 to 0.88 µM. Its selectivity toward malignant phenotype was confirmed on non-cancer V79-4 cells, and it was safe in vivo, for zebrafish embryos in doses up to 1 µM. Violacein induced apoptosis and affected the migratory potential of OS and RMS cells. It was found on the surfaces of tested cells. Regarding the mechanism of action, violacein acted on OS and RMS cells independently of oxidative signaling, as judged by no increase in intracellular ROS generation and no lipid peroxidation. CONCLUSION: Our study provided further evidence that reinforces the potential of violacein as an anticancer agent and candidate to consider for improvement of the effectiveness of traditional OS and RMS therapies.

Metagenomic Analysis of Bacterial Community and Isolation of Representative Strains from Vranjska Banja Hot Spring, Serbia.

The hot spring Vranjska Banja is the hottest spring on the Balkan Peninsula with a water temperature of 63-95 °C and a pH value of 7.1, in situ. According to the physicochemical analysis, Vranjska Banja hot spring belongs to the bicarbonated and sulfated hyperthermal waters. The structures of microbial community of this geothermal spring are still largely unexplored. In order to determine and monitor the diversity of microbiota of the Vranjska Banja hot spring, a comprehensive culture-independent metagenomic analysis was conducted in parallel with a culture-dependent approach for the first time. Microbial profiling using amplicon sequencing analysis revealed the presence of phylogenetically novel taxa, ranging from species to phyla. Cultivation-based methods resulted in the isolation of 17 strains belonging to the genera Anoxybacillus, Bacillus, Geobacillus, and Hydrogenophillus. Whole-genome sequencing of five representative strains was then performed. The genomic characterization and OrthoANI analysis revealed that the Vranjska Banja hot spring harbors phylogenetically novel species of the genus Anoxybacillus, proving its uniqueness. Moreover, these isolates contain stress response genes that enable them to survive in the harsh conditions of the hot springs. The results of the in silico analysis show that most of the sequenced strains have the potential to produce thermostable enzymes (proteases, lipases, amylases, phytase, chitinase, and glucanase) and various antimicrobial molecules that can be of great importance for industrial, agricultural, and biotechnological applications. Finally, this study provides a basis for further research and understanding of the metabolic potential of these microorganisms.

Isolation, Characterization, Genome Analysis and Host Resistance Development of Two Novel Lastavirus Phages Active against Pandrug-Resistant Klebsiella pneumoniae.

Klebsiella pneumoniae is a global health threat and bacteriophages are a potential solution in combating pandrug-resistant K. pneumoniae infections. Two lytic phages, LASTA and SJM3, active against several pandrug-resistant, nosocomial strains of K. pneumoniae were isolated and characterized. Their host range is narrow and latent period is particularly long; however, their lysogenic nature was refuted using both bioinformatic and experimental approaches. Genome sequence analysis clustered them with only two other phages into the new genus Lastavirus. Genomes of LASTA and SJM3 differ in only 13 base pairs, mainly located in tail fiber genes. Individual phages, as well as their cocktail, demonstrated significant bacterial reduction capacity in a time-dependent manner, yielding up to 4 log reduction against planktonic, and up to 2.59 log on biofilm-embedded, cells. Bacteria emerging from the contact with the phages developed resistance and achieved numbers comparable to the growth control after 24 h. The resistance to the phage seems to be of a transient nature and varies significantly between the two phages, as resistance to LASTA remained constant while resensitization to SJM3 was more prominent. Albeit with very few differences, SJM3 performed better than LASTA overall; however, more investigation is needed in order to consider them for therapeutic application.

Antioxidant and cytotoxic activity of red wine after in vitro simulated digestion in the presence of complex food matrix.

The beneficial effect of moderate wine consumption is attributed to its micronutrients, especially polyphenols and largely depends on the digestion process. This work aimed to examine the influence of in vitro simulated digestion in the presence of complex food matrix on antioxidant and cytotoxic activity of red wine. The obtained results showed that total phenolic content of wine sample after in vitro digestion was higher compared to undigested wine, while the antioxidant activity of these samples was similar before and after digestion. Furthermore, it has been noticed that digested wine showed cytotoxic activity on SKBR3 breast adenocarcinoma cells near 20% after 72 h of treatment. This pioneering study that examined biological potential of in vitro digested wine in the presence of complex food matrix indicate that antioxidant and cytotoxic activity of red wine is preserved after digestion.

Bioaccessibility of Phenolic Compounds and Antioxidant Properties of Goat-Milk Powder Fortified with Grape-Pomace-Seed Extract after In Vitro Gastrointestinal Digestion.

This study deals with the evaluation of the bioaccessibility and antioxidant properties of phenolic compounds from heat-treated skim goat-milk powder fortified with grape-pomace-seed extract, after in vitro gastrointestinal digestion. Ultra-high performance liquid chromatography coupled to diode array detection and mass spectrometry (UHPLC-DAD MS/MS) analysis confirmed the abundant presence of phenolic acids and flavan-3-ols in the grape-pomace-seed extract (SE) and heat-treated skim goat-milk/seed-extract powder (TME). After in vitro digestion of TME powder and recovery of total quantified phenolics, flavan-3-ols and phenolic acids were 18.11%, 24.54%, and 1.17%, respectively. Low recovery of grape-pomace-seed phenolics indicated strong milk protein-phenolic interactions. Electrophoretic analysis of a soluble fraction of digested heat-treated skim goat milk (TM) and TME samples showed the absence of bands originating from milk proteins, indicating their hydrolysis during in vitro gastrointestinal digestion. The digested TME sample had better antioxidant properties in comparison to the digested TM sample (except for the ferrous ion-chelating capacity, FCC), due to the presence of bioaccessible phenolics. Taking into account the contribution of the digestive cocktail, digested TME sample had lower values of total phenolic content (TPC), in vitro phosphomolybdenum reducing capacity (TAC) and ferric reducing power (FRP), compared to the undigested TME sample. These results could be attributed to low recovery of phenolic compounds. TME powder could be a good carrier of phenolics to the colon; thus, TME powder could be a promising ingredient in the formulation of functional food.

Coriolus versicolor Mushroom Grown on Selenium-Rich Zeolitic Tuff as a Potential Novel Food Supplement.

Research background: In the recent years, considerable attention has been given to selenium status since its deficiency is linked with various disorders and affects at least 13% of world population. Additionally, mushrooms are known to possess pronounced capacity for absorption of various micronutrients, including Se, from soil/substrate. Here, we investigate the possibility of using Se-rich zeolitic tuff as a supplement for production of selenized mushroom. Furthermore, the impact of the enrichment on the activity of antioxidant enzymes and biological potential of Coriolus versicolor medicinal mushroom is studied. Experimental approach: Se(IV)- and Se(VI)-modified natural zeolitic tuff from the Serbian deposit Zlatokop was used as supplement for mushroom cultivation. To examine the effectiveness of selenium enrichment, we determined total selenium with inductively coupled plasma mass spectrometry (ICP-MS), together with the activity of antioxidant enzymes in fresh fruiting bodies and biological potential of methanolic extracts. Antioxidant activity was evaluated using the appropriate tests for: inhibition of lipid peroxidation, DPPH free radical scavenging assay, Fe(III)-reducing antioxidant power assay and ability of chelating Fe2+ ions. The antibacterial activity against foodborne pathogens was measured by broth microdilution assay. Additionally, chemical composition of the prepared extracts was studied using UV-Vis and Fourier transform infrared (FTIR) spectroscopy. Results and conclusions: Content of selenium detected in biofortified C. versicolor was even 470 times higher than in control on dry mass basis ((140.7±3.8) vs (0.3±0.1) µg/g), proving that Se-rich zeolitic tuff is an excellent supplement for mushroom production. Furthermore, the results of monitoring the activity of antioxidant enzymes revealed that most of the Se-enriched mushrooms exhibited higher superoxide dismutase (SOD) and catalase (CAT) and lower glutathione peroxidase (GSH-Px) activities than control. Due to higher amounts of enzymes, which can quickly catalyze the reduction of superoxide radicals, the quality of selenium-enriched mushrooms is preserved for a longer period of time. Investigation of biological potential indicated that Se-enriched mushroom methanolic extracts, generally, expressed enhanced antioxidant properties. Additionally, extracts showed antibacterial activity against all tested pathogenic microorganisms. Novelty and scientific contribution: Cultivation of mushrooms on Se-enriched zeolitic tuff is a new technological approach for obtaining Se-fortified food/supplements with enhanced antioxidant and antibacterial activities.

Assessment of asbestos and asbestos waste quantity in the built environment of transition country.

Managing asbestos streams in developing and transition countries is particularly challenging. Deficiencies are often present for adequate procedures for the management of asbestos waste; solid quality data or databases on the quantities of asbestos production and usage are missing and asbestos inventories or the registry of asbestos-related diseases following European Union (EU) or other regulations are not in place. This paper aims to develop a model for determining and assessing the quantity of asbestos in the built environment of a transition country. Quantities of asbestos products and life expectancy of those products were assessed to develop a model that forecasts flows and stocks of asbestos products and wastes. The overall objective is to evaluate the model and show the manifestation of asbestos in the waste stream in a case study on a country with a transition economy, such as Serbia. Results show that total quantities of asbestos fibre consumption are approximately 0.5 million tonnes; the largest amount of waste generation is expected in the 2020s. Therefore, it is necessary to prepare for the forthcoming quantities of waste by improving legal procedures, implementation of existing regulations, and provision of economic resources. An important link for the adequate management of asbestos waste is to raise public awareness of the dangers and importance of proper and timely disposal of asbestos products.

WEEE treatment and system management in Italy and Serbia: A comparison.

Waste electrical and electronic equipment (WEEE) represent one of the growing waste streams in Europe, whose content of hazardous substances and valuable materials focus on how it is handled. However, there are differences between countries regarding e-waste management system in Europe. This paper analyses and presents data regarding the e-waste management system in Italy, one of the founding countries of the EU and Serbia - EU candidate country. Within this work, the following aspects were considered: legislative framework and EU directive goals in terms of e-waste, institutional setup, collection system, and existing recycling and treatment technologies. In addition, material flow analysis is used to model mass balance within WEEE treatment plants in both countries. Finally, through assessment and comparison of current systems in both countries, problems and shortages of Serbia's e-waste management system are addressed.

Bee pollen powder as a functional ingredient in frankfurters.

The objective of this study was to determine whether the addition of different pollen powder concentrations (0.0, 0.5, 1.0 and 1.5 g/100 g) to frankfurters had an influence on antioxidant potential and oxidative changes during storage, without detrimental effect on the quality of sausages. After cold storage of frankfurters, significant (P < 0.05) reductions of psychrotrophic bacteria populations were achieved with higher amounts of pollen (1.0 and 1.5 g/100 g). Good antioxidant properties and maintained TBARS values were accomplished by incorporating pollen into the frankfurters. In terms of quality parameters, statistically significant changes were obtained regarding the color, but sensory characteristics of the products were not disturbed. Also, the incorporation of pollen did not cause changes in terms of texture profile analyses of frankfurters. It can be concluded that the natural component, bee pollen powder, can be used as an antioxidant in frankfurter formulations, but further research is needed to estimate whether it can be an adequate replacement for synthetic antioxidants.

Determination of the composition and properties of PET bottles: Evidence of the empirical approach from Perm, Russia.

Efficient collection systems and information about the characteristics and quality of collected secondary plastic waste flows are of fundamental importance for the development of circular economies. In order to assess the effectiveness of the implementation of separate collection systems for plastic packaging, especially polyethylene terephthalate (PET) bottles, characteristic of the collected PET bottles in street mesh containers were studied in the city of Perm, Russia. The share of extraneous fractions was assessed and differentiation was carried out by volume, type of product, label presence, shape, content of solid and liquid impurities and colour. These results indicate that PET composition in different seasons is very similar, despite the assumption that the consumption of PET bottles in the spring and autumn seasons varies. In the mesh containers, up to 34% of the items were foreign objects, considering that only PET bottles should be collected. In each dimensional flow of PET bottles, the proportion of transparent bottles prevailed; it ranged from 31% to 70%. Based on the results of the experiment, almost all PET bottle categories had a standard shape, except packaging for food products and household chemicals, in which 26-27% of PET bottles had a non-standard shape. The results about charactersitic of source-separated PET bottles are fundamental for goal-oriented design and implementation of collection, recycling technologies, secondary separation facilities, the economics of recycling intitatives and reverse vending machines for collecting materials.

Polyphenol bioaccessibility and antioxidant properties of in vitro digested spray-dried thermally-treated skimmed goat milk enriched with pollen.

The aim of research was to determine polyphenols bioaccessibility and antioxidant properties of thermally-treated skimmed goat milk enriched with sunflower bee-collected pollen through in vitro digestion. HPLC analysis confirmed that pollen-enriched milk contained flavonols as the main phenolic fraction (80.7-76.2%) followed by phenolic acids (14.2-17.4%). Among individual compounds quercetin-3-O-glucoside (155.1-197.2 µg/L) and p-coumaric acid (29.5-30.7 µg/L) were the main quantified flavonols and phenolic acids, respectively. After digestion of milk/pollen sample, total polyphenols recovery was 30.71% with higher phenolic acids recovery (40.1%) compared to flavonols (28.3%) indicating strong interactions between caprine milk casein micelles and pollen polyphenols. Applied antioxidant assays (phosphomolybdenum, ABTS•+scavenging activity and ferrous-ion-chelating capacity) have confirmed complexity of prepared product- it had high ability to quench ABTS•+ radicals and to form chelating complexes with Fe2+ ions. Digestion provoked 20% reduction in total antioxidant capacity compared to the initial sample. TTSG milk/pollen powder could be good functional ingredient.

The large plasmidome of Lactococcus lactis subsp. lactis bv. diacetylactis S50 confers its biotechnological properties.

Plasmids are autonomous episomally replicating genetic elements, which carry backbone genes important for the replication and maintenance within their host, and accessory genes that might confer an advantage to their host under specific selective pressure in its ecological niche. The genome of dairy isolate L. lactis subsp. lactis bv. diacetylactis S50 was sequenced using the PacBio SMRT Cell Seq-RSII platform and revealed to possess one of the largest plasmidomes among L. lactis strains studied so far, harboring six plasmids: pS6 (5553 bp), pS7a (7308 bp), pS7b (7266 bp), pS19 (19,027 bp), pS74 (74,256 bp) and pS127 (127,002 bp) in total representing 8.9% of genome size (240,412 bp). Based on predicted plasmid replication proteins and origins it appears that all six plasmids replicate via the theta-type mechanism. The two the largest plasmids (pS74 and pS127), carry a number of genes known to be important for growth and survival in the dairy environment. These genes encode technological functions such as bacteriocin production, protein degradation, magnesium and cobalt/nickel transporters, selenium binding, exopolysaccharides (EPS) production, bacteriophage and stress resistance. Beside genes for replication, the small plasmids (pS6, pS7a, pS7a, and pS19) also carry genes important for mobilization and host survival such as type I restriction-modification (R-M) system, metal transporters, enzymes and transcriptional regulators. All plasmids in S50 strain are mobilizable, containing an oriT sequences, while pS127 is self-conjugative and allows for mobilization of the other plasmids. Small plasmids are prone to structural and segregational instability, while pS127 appeared to be segregationally stable thanks to the possession of two partition systems. The main characteristic of plasmid pS74 is EPS production, while plasmid pS127 is characterized by proteinase and multiple bacteriocins, tra locus, phage abortive systems and metal transporters. In addition to LcnA and LcnB, plasmid pS127 encodes several bacteriocin-pheromone molecules and a new bacteriocin named LcnS50, with narrow spectrum of action limited to lactococci, that has been successfully cloned and heterologously expressed.

Antioxidant and Antiproliferative Activity of Allium ursinum and Their Associated Microbiota During Simulated in vitro Digestion in the Presence of Food Matrix.

In this study, for the first time, the comprehensive analysis of antiproliferative and antioxidant activities of ramson, followed by the analysis of its associated microbiota and health-promoting effects of lactic acid bacteria (LAB), was performed. Ramson (Allium ursinum) is recognized as a medicinal plant with a long history of use in traditional medicine due to its antimicrobial and antioxidant activity. In this study the influence of in vitro gastrointestinal digestion on the cytotoxic activity of A. ursinum extracts against human malignant cell lines was demonstrated. Seven sulfur compounds, the degradation products of thiosulfinates, including diallyl disulfide were shown to inhibit proliferation of malignant cells by inducing accumulation within G2/M phase as well as to induce apoptosis through activation of caspase-3 and mitochondrial signaling pathway. Further, the A. ursinum microbiota, particularly LAB with potential probiotic effects, was analyzed by culture-dependent method and culture-independent method [denaturing gradient gel electrophoresis (DGGE)]. The obtained results revealed that the most abundant genera were Streptococcus, Lactobacillus, and Bacillus. The Lactobacillus genus was mainly represented by L. fermentum. The pulsed-field gel electrophoresis (PFGE) analysis revealed the presence of two PFGE pulsotypes. The probiotic potential of the strain L. fermentum BGSR163 belonging to PFGE pulsotype 1 and the strain L. fermentum BGSR227 belonging to the PFGE pulsotype 2 was characterized. The results revealed that both strains are safe for human use, successfully survive the simulated gastrointestinal conditions, have potential to transiently colonize the gastrointestinal tract (GIT) and have a protective immunomodulatory effect, inducing the production of proinflammatory cytokine IL17 and regulatory cytokine IL10, while decreasing the production of proinflammatory cytokine IFN-γ. In conclusion, the results of this study suggest that consumption of A. ursinum might have health-promoting properties, including anticancer effects, while L. fermentum strains isolated from A. ursinum leaves could be used as probiotics for human consumption.

Burkholderia cepacia YtnP and Y2-aiiA lactonases inhibit virulence of Pseudomonas aeruginosa via quorum quenching activity.

Burkholderia cepacia is well known as the causative agent of infections in humans where often shares niche with other pathogens, like Pseudomonas aeruginosa. Clinical isolate Burkholderia sp. BCC4135 was selected due to its strong quorum quenching (QQ) activity. Whole genome sequencing unveiled this isolate as B. cepacia with unique sequence type ST1485 and a myriad of genes belonging to resistome and virulome. Two QQ lactonases YtnP and Y2-aiiA originated from B. cepacia BCC4135 were cloned, expressed, and functionally characterized. They were active against a broad substrate spectrum of the N-acyl-homoserine lactones (AHLs). The YtnP lactonase was inactive, while Y2-aiiA was active against N-tetradecanoyl-dl-homoserine lactone (C14-HSL) which could imply the difference in their biological roles from the aspect of its quorum sensing (QS) autoregulation and interference with the QS systems of bacteria residing within the same niche. Both YtnP and Y2-aiiA were able to attenuate virulence potential of P. aeruginosa MMA83 clinical isolate declining its biofilm formation and virulence factors production. B. cepacia BCC4135 lactonases interfered with the las, rhl, and even pqs QS circuit of P. aeruginosa MMA83 transcription and the effect of combined enzymes was even more prominent. B. cepacia BCC4135 also employs the CepI/R QS system for governing its own virulence traits and possibly self-regulates the QQ/QS network through the different expression and activity of YtnP and/or Y2-aiiA. Our findings pointed out that BCC4135 lactonases could be exploited as an effective antivirulence drugs against P. aeruginosa and gave us a new insight into B. cepacia QQ/QS machinery.

Material efficiency to measure the environmental performance of waste management systems: A case study on PET bottle recycling in Austria, Germany and Serbia.

Material efficiency measures, such as recycling rates, are often used to set circular economy targets to achieve higher resource efficiency and lower environmental impact. The aim of this study was to identify material efficiency indicators suitable to reflect the environmental performance of waste and recycling systems using PET bottle waste management in three European countries with diverse waste management structures and recycling performance levels. Material flow analysis and life cycle assessment were performed to assess the material efficiency and environmental impacts of each system as a basis to analyze the relation between these two dimensions. PET bottle waste generation was 5.4 kg/person and year (pa) in Austria in 2013, 6.0 kg/pa in Germany in 2017 and 6.9 kg/pa in Serbia in 2015. Out of this waste flow 41%, 91%, and 11% were directed into PET recyclate in Austria, Germany and Serbia, respectively. For all systems, higher material efficiency translated into lower environmental impact and vice versa. However, linear regression analysis between different material efficiency indicators and environmental impacts showed that indicators targeted at actual recycling, specifically at closed loop, were better suited to reflect environmental performance than input-based indicators. Therefore, whenever data are available, output-based quality-related indicators should be used to measure the material efficiency of waste and resource systems because they correlate best with the goals of increasing resource efficiency and decreasing environmental impacts.