Biodegradation of untreated plasticizers-free linear low-density polyethylene films by marine bacteria.

Polyethylene significantly contributes to marine plastic pollution. This study focuses on isolating bacteria from sea water and microplastic samples collected from the Tyrrhenian Sea and evaluating their ability to degrade virgin plasticizers-free linear low-density polyethylene (LLDPE) films. The isolates grew on the plastic film under aerobic conditions in shaken flasks leading to LLDPE mass losses of up to 2.597 ± 0.971 % after 60 days incubation. Biofilm formation on the film surface was confirmed by adhered protein quantification while film surface erosion and appearance of functional groups were revealed using SEM and FTIR analyses confirming biodegradation capabilities especially for isolates Bacillus velezensis MT9, Vreelandella venusta MT1 and Vreelandellatitanicae MT11. This is the first report on the biodegradation of plasticizers-free non pretreated LLDPE films by marine Bacillus sp. and Vreelandella sp.; most of the LLDPE biodegradation studies have been so far performed on plasticizer containing, pre-treated, or naturally weathered films.

Chemical-physical parameters and microbial community changes induced by electrodes polarization inhibit PCB dechlorination in a marine sediment.

Microbial reductive dechlorination of organohalogenated pollutants is often limited by the scarcity of electron donors, that can be overcome with microbial electrochemical technologies (METs). In this study, polarized electrodes buried in marine sediment microcosms were investigated to stimulate PCB reductive dechlorination under potentiostatic (-0.7 V vs Ag/AgCl) and galvanostatic conditions (0.025 mA·cm-2-0.05 mA·cm-2), using graphite rod as cathode and iron plate as sacrificial anode. A single circuit and a novel two antiparallel circuits configuration (2AP) were investigated. Single circuit polarization impacted the sediment pH and redox potential (ORP) proportionally to the intensity of the electrical input and inhibited PCB reductive dechlorination. The effects on the sediment's pH and ORP, along with the inhibition of PCB reductive dechlorination, were mitigated in the 2AP system. Electrodes polarization stimulated sulfate-reduction and promoted the enrichment of bacterial clades potentially involved in sulfate-reduction as well as in sulfur oxidation. This suggested the electrons provided were consumed by competitors of organohalide respiring bacteria and specifically sequestered by sulfur cycling, which may represent the main factor limiting the applicability of METs for stimulating PCB reductive dechlorination in marine sediments.

Site-specific response of sediment microbial community to supplementation of polyhydroxyalkanoates as biostimulants for PCB reductive dechlorination.

The use of biodegradable plastics is constantly raising, increasing the likeliness for these polymers to end up in the environment. Environmental applications foreseeing the intentional release of biodegradable plastics have been also recently proposed, e.g., for polyhydroxyalkanoates (PHAs) acting as slow hydrogen releasing compounds to stimulate microbial reductive dehalogenation processes. However, the effects of their release into the environment on the ecosystems still need to be thoroughly explored. In this work, the use of PHAs to enhance the microbial reductive dechlorination of polychlorobiphenyls (PCBs) and their impact on the metabolic and compositional features of the resident microbial community have been investigated in laboratory microcosms of a polluted marine sediment from Mar Piccolo (Taranto, Italy), and compared with recent findings on a different contaminated marine sediment from Pialassa della Baiona (Ravenna, Italy). A decreased biostimulation efficiency of PHAs on PCBs reductive dechlorination was observed in the sediment from Mar Piccolo, with respect to the sediment from Pialassa della Baiona, suggesting that the sediments' physical-chemical characteristics and/or the biodiversity and composition of its microbial community might play a key role in determining the outcome of this biostimulation strategy. Regardless of the sediment origin, PHAs were found to have a specific and pervasive effect on the sediment microbial community, reducing its biodiversity, defining a newly arranged microbial core of primary degraders and consequently affecting, in a site-specific way, the abundance of subdominant bacteria, possibly cross-feeders. Such potential to dramatically change the structure of autochthonous microbial communities should be carefully considered, since it might have secondary effects, e.g., on the natural biogeochemical cycles.

Front-of-pack labels: "Directive" versus "informative" approaches.

Front-of-pack labels (FOPLs) aim at communicating to consumers the health value of food items in support of public health policies. Two main types can be discerned: directive and semidirective FOPLs using color schemes (e.g., Nutri-Score) and informative FOPLs (e.g., Nutrinform Battery). Directive approaches tend to show a "wear-out effect" and, additionally, they tend to have various underlying conceptual problems. Usually, their nutritional scores are calculated using changing, arbitrary algorithms and involve a reductionist set of parameters of debatable validity. Thus, they overstate the effects of selected nutritional factors, such as saturated fat and energy, while overlooking the food matrix and the more holistic aspects of nourishment. Moreover, they do not reflect the portion that is consumed, ignore the preparation steps at home, and fail to serve as a useful basis for composing a healthy diet. Also, so long as the nutritional formulations match the algorithmic standards, they tend to allow ultra-processed products. Thus, this might confuse and mislead consumers. Overconfidence in green-colored labels could even result in unbalanced dietary choices, whereas avoidance of red products may eliminate certain foods from the diet that are rich in essential nutrients (e.g., cheese), leading to opposite results than aimed for. The latter is particularly relevant to vulnerable populations, such as the young, pregnant women, and older adults, or for individuals with specific needs. Taken together, directive FOPLs such as Nutri-Score contradict the declared intent of the European Commission to empower consumers to undertake healthy and balanced diets based on easily accessible and robust information. Although informative systems usually also keep the focus on a few selected nutritional parameters, they have are less paternalizing and obviate the need to classify foods as healthy or unhealthy. They also focus attention on the individual portions that are consumed (even if the definition of portion size remains contentious). Given the importance of dietary patterns, rather than individual foods or nutrients, directiveFOPLs of the Nutri-Score type represent a regretful case of nutritionism. Finally, attempts to associate the adoption of a FOPL with an improvement in the health status are few and mainly applied in virtual settings; none of which are longitudinal, nor have they been able to identify a causal link.

Effect of polyhydroxyalkanoates on the microbial reductive dechlorination of polychlorinated biphenyls and competing anaerobic respirations in a marine microbial culture.

The effect of polyhydroxyalkanoates (PHAs) with different composition on the reductive dechlorination activity of a polychlorinated biphenyls (PCBs) dechlorinating marine microbial community and on the activity of sulfate-reducing (SRB) and methanogenic bacteria (MB), were investigated in marine sediment microcosms and compared with the main monomer, 3-hydroxybutyric acid (3HB). Despite PHAs were fermented more slowly than 3HB, all electron donors stimulated constantly sulfate-reduction, methanogenesis and, only transiently, PCB reductive dechlorination. No relevant differences were observed with different compositions of PHAs. According to electron balances, the majority of the supplied electrons (50 %) were consumed by SRB and to less extent by MB (9-31 %), while a small percentage (0.01 %) was delivered to OHRB. In the studied conditions PHAs were confirmed as potential slow­hydrogen releasing compounds in marine environment but their fermentation rate was sufficiently high to mainly stimulate the competitors of organohalide respring bacteria for electron donors.

Bacterial colonization dynamics of different microplastic types in an anoxic salt marsh sediment and impact of adsorbed polychlorinated biphenyls on the plastisphere.

Plastic debris dispersed into the environment provide a substrate for microbial colonization, constituting a new human-made ecosystem called "plastisphere", and altering the microbial species distribution in aquatic, coastal and benthic ecosystems. The study aims at exploring the interaction among microplastics (MPs) made of different polymers, a persistent organic contaminant (polychlorinated biphenyls, PCBs), and the environmental microbial communities, in an anoxic marine sediment. Plastic pellets were incubated in the field in a salt marsh anoxic sediment, to observe the stages of plastisphere formation, by quantitative PCR and 16S rRNA gene sequencing, and PCB dechlorination activity on the MPs surface. Microbes from the sediment rapidly colonized the different microplastics types, with PVC recruiting a peculiar community enriched in sulfate-reducing bacteria. The composition of the plastisphere varied along the 1-year incubation possibly in response either to warmer temperatures in spring-summer or to microhabitat's changes due to the progressive plastic surface weathering. Even if PCB contaminated MPs were able to recruit potentially dehalogenating taxa, actual dechlorination was not detectable after 1 year. This suggests that the concentration of potentially dehalorespiring bacteria in the natural environment could be too low for the onset of the dechlorination process on MP-sorbed contaminants. Our study, which is among very few available longitudinally exploring the plastisphere composition in an anoxic sediment context, is the first exploring the fate and possible biodegradation of persistent organic pollutants sorbed on MPs reaching the seafloor.

Enzymatic Degradation of the Most Common Aliphatic Bio-Polyesters and Evaluation of the Mechanisms Involved: An Extended Study.

Commercial hydrolytic enzymes belonging to different subclasses (several lipases, proteinase k, cutinase) were investigated for their ability to degrade different aliphatic polyesters, i.e., poly(butylene succinate) (PBS), poly(butylene succinate-co-adipate) (PBSA), two poly(caprolactone), having two different molecular weights, poly(lactic acid) (PLA) and poly(propylene carbonate) (PPC). The enzyme screening was first carried out by investigating the capacity of fully degrading the target polymers in 24 h, then weight loss measurements of selected polyesters and target enzymes were performed. Solid residues after enzyme degradation were characterized by proton nuclear magnetic resonance (1H NMR), gel permeation chromatography (GPC), infrared spectroscopy (FT-IR), differential scanning calorimetry (DSC) and thermogravimetry (TGA). Liquid fractions were studied via GPC, 1H NMR and high-performance liquid chromatography (HPLC). PCL and PBSA were found to be the most biodegradable polyesters, under the conditions used in this study. PBS was fully degraded only by cutinase, whereas none of the tested enzymes were able to completely degrade PLA and PPC, in the conditions assessed here. Cutinase exhibited the highest hydrolytic activity on PBSA, while lipase from Candida sp. (CALB) on low molecular weight PCL. Chemical analyses on residual solids showed that the enzymatic degradation occurred homogeneously from the surface through an erosion mechanism and did not significantly affect the macromolecular structure and thermal stability. Cleaving action mode for each enzyme (endo- and/or exo-type) on the different polyesters were also proposed based on the evaluation of the degradation products in the liquid fraction.

A Multidisciplinary Perspective of Ultra-Processed Foods and Associated Food Processing Technologies: A View of the Sustainable Road Ahead.

Ultra-processed foods (UPFs) are negatively perceived by part of the scientific community, the public, and policymakers alike, to the extent they are sometimes referred to as not "real food". Many observational surveys have linked consumption of UPFs to adverse health outcomes. This narrative synthesis and scientific reappraisal of available evidence aims to: (i) critically evaluate UPF-related scientific literature on diet and disease and identify possible research gaps or biases in the interpretation of data; (ii) emphasize the innovative potential of various processing technologies that can lead to modifications of the food matrix with beneficial health effects; (iii) highlight the possible links between processing, sustainability and circular economy through the valorisation of by-products; and (iv) delineate the conceptual parameters of new paradigms in food evaluation and classification systems. Although greater consumption of UPFs has been associated with obesity, unfavorable cardiometabolic risk factor profiles, and increased risk for non-communicable diseases, whether specific food processing techniques leading to ultra-processed formulations are responsible for the observed links between UPFs and various health outcomes remains elusive and far from being understood. Evolving technologies can be used in the context of sustainable valorisation of food processing by-products to create novel, low-cost UPFs with improved nutritional value and health potential. New paradigms of food evaluation and assessment should be funded and developed on several novel pillars-enginomics, signalling, and precision nutrition-taking advantage of available digital technologies and artificial intelligence. Research is needed to generate required scientific knowledge to either expand the current or create new food evaluation and classification systems, incorporating processing aspects that may have a significant impact on health and wellness, together with factors related to the personalization of foods and diets, while not neglecting recycling and sustainability aspects. The complexity and the predicted immense size of these tasks calls for open innovation mentality and a new mindset promoting multidisciplinary collaborations and partnerships between academia and industry.

The role of biotechnology in the transition from plastics to bioplastics: an opportunity to reconnect global growth with sustainability.

Building new value chains, through the valorization of biomass components for the development of innovative bio-based products (BBPs) aimed at specific market sectors, will accelerate the transition from traditional production technologies to the concept of biorefineries. Recent studies aimed at mapping the most relevant innovations undergoing in the field of BBPs (Fabbri et al. 2019, Final Report of the Task 3 BIOSPRI Tender Study on Support to R&I Policy in the Area of Bio-based Products and Services, delivered to the European Commission (DG RTD)), clearly showed the dominant position played by the plastics sector, in which new materials and innovative technical solutions based on renewable resources, concretely contribute to the achievement of relevant global sustainability goals. New sustainable solutions for the plastic sector, either bio-based or bio-based and biodegradable, have been intensely investigated in recent years. The global bioplastics and biopolymers market size is expected to grow from USD 10.5 billion in 2020 to USD 27.9 billion by 2025 (Markets and Markets, 2020, Bioplastics & Biopolymers Market by Type (Non-Biodegradable/Bio-Based, Biodegradable), End-Use Industry (Packaging, Consumer Goods, Automotive & Transportation, Textiles, Agriculture & Horticulture), Region - Global Forecast to 2025), and this high growth is driven primarily by the growth of the global packaging end-use industry. Such relevant opportunities are the outcomes of intensive scientific and technological research devoted to the development of new materials with selected technical features, which can represent feasible substitutes for the fossil-based plastic materials currently used in the packaging sectors and other main fields. This article offers a map of the latest developments connected to the plastic sector, achieved through the application of biotechnological routes for the preparation of completely new polymeric structures, or drop-in substitutes derived from renewable resources, and it describes the specific role played by biotechnology in promoting and making this transition faster.

The bioeconomy in Italy and the new national strategy for a more competitive and sustainable country.

Italy has the third largest bioeconomy in Europe (€330 billion annual turnover, 2 million employees), making it a core pillar of the national economy. Its sectors of excellence are food and biobased products, and it is a consistent presence in research and innovation projects funded by the EU Horizon 2020 programme (Societal Challenges 2) and the European Public Private Partnership "Biobased industry" (BBI-JU). The bioeconomy reduces dependence on fossil fuels and finite materials, loss of biodiversity and changing land use. It contributes to environmental regeneration, spurs economic growth and supports jobs in rural, coastal and abandoned industrial areas, leveraging local contexts and traditions. In 2017 the Italian government promoted the development of a national Bioeconomy Strategy (BIT), recently updated (BIT II) to interconnect more efficiently the pillars of the national bioeconomy: production of renewable biological resources, their conversion into valuable food/feed, biobased products and bio-energy, and transformation and valorization of bio-waste streams. BIT II aims to improve coordination between Ministries and Italian regions in alignment of policies, regulations, R&I funding programmes and infrastructures investment. The goal is a 15 % increase in turnover and employment in the Italian bioeconomy by 2030. Based on Italy's strategic geopolitical position in the Mediterranean basin, BIT II also includes actions to improve sustainable productivity, social cohesion and political stability through the implementation of bioeconomy strategies in this area. This paper provides an insight into these strategies and discusses the strengths and weaknesses of the sectors involved and the measures, regulatory initiatives and monitoring actions undertaken.

Mediterranean Sea bacteria as a potential source of long-chain polyunsaturated fatty acids.

Long-chain polyunsaturated fatty acids (LC-PUFAs), including EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid) are important nutritional ingredients in fish feed. So far, fish oil has been used as a main source of LC-PUFAs; however, the limited global supply of fish oil is not able to meet the demand of the growing aquaculture sector. Hence, sustainability of aquaculture industry could be supported by searching alternative sources of these compounds. Marine microorganisms represent a sustainable and stable supply source of LC-PUFAs. A collection of 209 bacterial isolates obtained from sediment samples recovered in the Mediterranean Sea was screened in order to select new LC-PUFAs producers. Among 95 putative producers selected based on colourimetric screening, 31 quickly growing were selected for further studies. The detection of LC-PUFAs was confirmed from 15 isolates belonging to the genera Marinobacter, Halomonas and Thalassospira by GC-FID analysis. Among them, the isolate Marinobacter sp. G16.20 was found to be a potentially high LC-PUFA producer exhibiting relatively high levels of DHA in particular (maximum productivity of 1.85 ± 0.371 mg/g, representing 45.89% of the total fatty acids detected and identified). Microorganisms belonging to the genera reported in this study showed biotechnological traits interesting for their potential future application in aquaculture.

Biodegradation of polyvinyl chloride plastic films by enriched anaerobic marine consortia.

Plastics remarkably contribute to marine litter, which is raising serious concerns. Currently, little is known about the fate of most plastics entering the marine environment and their potential biodegradation rate and extent under anoxic conditions. In this work, biodegradation of polyvinyl chloride (PVC) films by consortia enriched from marine samples (litter and water) was evaluated in anaerobic microcosms. After 7 months, three microcosms showed dense biofilms on plastic surfaces, gravimetric weight losses up to 11.7 ± 0.6%, marked decreases in thermal stability and average molecular weight of the polymer, suggesting microbial attack towards polymer chains. After 24 months, further three consortia showed the same abilities. Microbial communities analyzed at month 24 included taxa closely related to those previously reported as halogenated organic compounds degraders. The study is the first report on PVC biodegradation by marine anaerobic microbes and provides insights on potential biodegradation of the plastic film introduced into the sea by native microbes.

Mineral-Doped Poly(L-lactide) Acid Scaffolds Enriched with Exosomes Improve Osteogenic Commitment of Human Adipose-Derived Mesenchymal Stem Cells.

Exosomes derived from mesenchymal stem cells are extracellular vesicles released to facilitate cell communication and function. Recently, polylactic acid (PLA), calcium silicates (CaSi), and dicalcium phosphate dihydrate (DCPD) have been used to produce bioresorbable functional mineral-doped porous scaffolds-through thermally induced phase separation technique, as materials for bone regeneration. The aim of this study was to investigate the effect of mineral-doped PLA-based porous scaffolds enriched with exosome vesicles (EVs) on osteogenic commitment of human adipose mesenchymal stem cells (hAD-MSCs). Two different mineral-doped scaffolds were produced: PLA-10CaSi-10DCPD and PLA-5CaSi-5DCPD. Scaffolds surface micromorphology was investigated by ESEM-EDX before and after 28 days immersion in simulated body fluid (HBSS). Exosomes were deposited on the surface of the scaffolds and the effect of exosome-enriched scaffolds on osteogenic commitment of hAD-MSCs cultured in proximity of the scaffolds has been evaluated by real time PCR. In addition, the biocompatibility was evaluated by direct-contact seeding hAD-MSCs on scaffolds surface-using MTT viability test. In both formulations, ESEM showed pores similar in shape (circular and elliptic) and size (from 10-30 µm diameter). The porosity of the scaffolds decreased after 28 days immersion in simulated body fluid. Mineral-doped scaffolds showed a dynamic surface and created a suitable bone-forming microenvironment. The presence of the mineral fillers increased the osteogenic commitment of hAD-MSCs. Exosomes were easily entrapped on the surface of the scaffolds and their presence improved gene expression of major markers of osteogenesis such as collagen type I, osteopontin, osteonectin, osteocalcin. The experimental scaffolds enriched with exosomes, in particular PLA-10CaSi-10DCPD, increased the osteogenic commitment of MSCs. In conclusion, the enrichment of bioresorbable functional scaffolds with exosomes is confirmed as a potential strategy to improve bone regeneration procedures.

Vascular Wall-Mesenchymal Stem Cells Differentiation on 3D Biodegradable Highly Porous CaSi-DCPD Doped Poly (α-hydroxy) Acids Scaffolds for Bone Regeneration.

Vascularization is a crucial factor when approaching any engineered tissue. Vascular wall-mesenchymal stem cells are an excellent in vitro model to study vascular remodeling due to their strong angiogenic attitude. This study aimed to demonstrate the angiogenic potential of experimental highly porous scaffolds based on polylactic acid (PLA) or poly-e-caprolactone (PCL) doped with calcium silicates (CaSi) and dicalcium phosphate dihydrate (DCPD), namely PLA-10CaSi-10DCPD and PCL-10CaSi-10DCPD, designed for the regeneration of bone defects. Vascular wall-mesenchymal stem cells (VW-MSCs) derived from pig thoracic aorta were seeded on the scaffolds and the expression of angiogenic markers, i.e. CD90 (mesenchymal stem/stromal cell surface marker), pericyte genes α-SMA (alpha smooth muscle actin), PDGFR-ß (platelet-derived growth factor receptor-ß), and NG2 (neuron-glial antigen 2) was evaluated. Pure PLA and pure PCL scaffolds and cell culture plastic were used as controls (3D in vitro model vs. 2D in vitro model). The results clearly demonstrated that the vascular wall mesenchymal cells colonized the scaffolds and were metabolically active. Cells, grown in these 3D systems, showed the typical gene expression profile they have in control 2D culture, although with some main quantitative differences. DNA staining and immunofluorescence assay for alpha-tubulin confirmed a cellular presence on both scaffolds. However, VW-MSCs cultured on PLA-10CaSi-10DCPD showed an individual cells growth, whilst on PCL-10CaSi-10DCPD scaffolds VW-MSCs grew in spherical clusters. In conclusion, vascular wall mesenchymal stem cells demonstrated the ability to colonize PLA and PCL scaffolds doped with CaSi-DCPD for new vessels formation and a potential for tissue regeneration.

Microbial colonization of different microplastic types and biotransformation of sorbed PCBs by a marine anaerobic bacterial community.

We investigated the colonization dynamics of different microplastic (MP) pellets, namely, polyethylene (PE), polyethylene terephthalate (PET), polystyrene (PS), polypropylene (PP) and polyvinyl chloride (PVC), either pristine or contaminated with polychlorinated biphenyls (PCBs), by an organohalide respiring marine microbial community and its biotransformation activity towards PCBs sorbed on MPs, in anaerobic laboratory microcosms of a marine sediment. All MPs were rapidly colonized by the microbial community within 2 weeks of incubation, when approximately 1010 16S rRNA gene copies cm-2 were detected on PVC, 109 copies cm-2 on PE, and 108 copies cm-2 on PET, PP and PS. A greater biofilm growth on PVC pellets than other MPs was confirmed by quantification of the reducing sugars of the EPS and biofilm staining with crystal violet. Illumina sequencing of the 16S rRNA genes and Principal Coordinate Analysis (PCoA) revealed that the biofilm community on MPs significantly differed from the sediment community, being enriched of chemoorganotrophic fermenting species, and was significantly affected by the type of polymer. The presence of sorbed PCBs did not significantly affect the overall community composition, and mainly resulted in the enrichment of Dehalococcoidia, i.e., of the organohalide respiring members of the community. Reductive dechlorination of PCBs sorbed to MPs was observed after 2 weeks of incubation, when the average number of chlorines per biphenyl molecule was reduced from 5.2 to 4.8-4.3, and was faster (35.2 ± 1.9 to 61.2 ± 5.8 µmol of Cl removed kgMP-1 week-1) than that of sediment-sorbed ones (33.9 ± 9.1 µmol of Cl removed kgsediment-1 week-1), which started only after 10 weeks of incubation. These data suggest that microbial colonization of contaminated MPs might change the composition of sorbed PCB mixtures and therefore the toxicity associated to PCB-polluted MPs.

Containment of a genetically modified microorganism by an activated sludge system.

The effectiveness of physical, chemical and biological barriers to the diffusion of genetically modified microorganisms (GMMs) to prevent their release into the environment is currently under scrutiny worldwide because of the associated potential ecological impacts. An industrial discharge of a non-sterilized fermentation broth containing GMM biomass into a conventional municipal wastewater treatment plant would deliver the GMMs into the activated sludge system process (ASSP). The present work aimed to model and evaluate the containment capability of a small ASSP (part of a 20,000 people equivalent municipal plant) in the event of receiving GMM biomass from a medium-small biotechnological plant dedicated to the production of polyhydroxyalkanoates (3000 t/year of biopolymer). An actual GMM (Pseudomonas putida KTOY06) was injected into a bench-scale ASSP (ASSPLab) in a quantity proportional to the relative dimensions of the plants mentioned. The experimental and model results indicated that the ASSP of the target municipal treatment plant would not be capable of holding back such a sudden input of GMM; 6 h after the discharge, 11-15 % of injected GMM cells were released through the clarified stream of the ASSPLab, with the rest being gradually released over time. Since the GMM employed did not exhibit any growth in the ASSPLab, its concentration in the clarified water stream would not represent a substantial risk of release into the environment if appropriate tertiary treatments were integrated. This study confirmed the necessity of a thorough risk assessment of biotechnological processes prior to their implementation.

Highly porous polycaprolactone scaffolds doped with calcium silicate and dicalcium phosphate dihydrate designed for bone regeneration.

Polycaprolactone (PCL), dicalcium phosphate dihydrate (DCPD) and/or calcium silicates (CaSi) have been used to prepare highly porous scaffolds by thermally induced phase separation technique (TIPS). Three experimental mineral-doped formulations were prepared (PCL-10CaSi, PCL-5CaSi-5DCPD, PCL-10CaSi-10DCPD); pure PCL scaffolds constituted the control group. Scaffolds were tested for their chemical-physical and biological properties, namely thermal properties by differential scanning calorimetry (DSC), mechanical properties by quasi-static parallel-plates compression testing, porosity by a standard water-absorption method calcium release, alkalinizing activity, surface microchemistry and micromorphology by Environmental Scanning electronic Microscopy (ESEM), apatite-forming ability in Hank Balanced Saline Solution (HBSS) by Energy Dispersive X-ray Spectroscopy (EDX) and micro-Raman, and direct contact cytotoxicity. All mineral-doped scaffolds released calcium and alkalinized the soaking medium, which may favor a good biological (osteogenic) response. ESEM surface micromorphology analyses after soaking in HBSS revealed: pure PCL, PCL-10CaSi and PCL-10CaSi-10DCPD kept similar surface porosity percentages but different pore shape modifications. PCL-5CaSi-5DCPD revealed a significant surface porosity increase despite calcium phosphates nucleation (p < 0.05). Micro-Raman spectroscopy detected the formation of a B-type carbonated apatite (Ap) layer on the surface of PCL-10CaSi-10DCPD aged for 28 days in HBSS; a similar phase (but of lower thickness) formed also on PCL-5CaSi-5DCPD and PCL; the deposit formed on PCL-10CaSi was mainly composed of calcite. All PCL showed bulk open porosity higher than 94%; however, no relevant brittleness was observed in the materials, which retained the possibility to be handled without collapsing. The thermo-mechanical properties showed that the reinforcing and nucleating action of the inorganic fillers CaSi and DCPD improved viscoelastic properties of the scaffolds, as confirmed by the increased value of storage modulus and the slight increase in the crystallization temperature for all the biomaterials. A detrimental effect on the mechanical properties was observed in samples with the highest amount of inorganic particles (PCL-10CaSi-10DCPD). All the scaffolds showed absence of toxicity, in particular PCL-10CaSi-10DCPD. The designed scaffolds are biointeractive (release biologically relevant ions), nucleate apatite, possess high surface and internal open porosity and can be colonized by cells, creating a bone forming osteoblastic microenvironment and appearing interesting materials for bone regeneration purposes.

Biodegradation of oil-based plastics in the environment: Existing knowledge and needs of research and innovation.

The production of synthetic oil-based plastics has led to the accumulation of huge amounts of the plastic waste in the environment, especially in the marine system, very often the final sink for many types of conventional wasted plastics. In particular, (micro)plastics account for the majority of litter items in the marine environment and a high percentage of such litter is originating from land sources. Attempts to mitigate the harmful effects of conventional plastics such as the development of novel management strategies together with the gradual substitution of them with biodegradable (bio)plastics are representing future solutions. However, high amounts of conventional plastics have been accumulating in the environment since several years. Although many studies reported on their potential biodegradation by microbes in and from terrestrial environments, very little is known about the biodegradability of these plastics in freshwater systems and only recently more reports on their biodegradation by marine microorganisms/in marine environment were made available. In this review, we first provide a summary of the approaches applied for monitoring and assessing conventional plastics biodegradation under defined conditions. Then, we reviewed historical and recent findings related to biodegradation of four major plastics produced in European Union (EU), i.e. Polyethylene, Polyvinyl Chloride, Polypropylene and Polystyrene, in terrestrial and aquatic environments and by pure and mixed microbial cultures obtained from them.

Biodegradation of mixture of plastic films by tailored marine consortia.

This work sheds light on the physicochemical changes of naturally weathered polymer surfaces along with changes of polymer buoyancy due to biofilm formation and degradation processes. To support the degradation hypothesis, a microcosm experiment was conducted where a mixture of naturally weathered plastic pieces was incubated with an indigenous pelagic community. A series of analyses were employed in order to describe the alteration of the physicochemical characteristics of the polymer (FTIR, SEC and GPC, sinking velocity) as well as the biofilm community (NGS). At the end of phase II, the fraction of double bonds in the surface of microbially treated PE films increased while changes were also observed in the profile of the PS films. The molecular weight of PE pieces increased with incubation time reaching the molecular weight of the virgin pieces (230,000 g mol-1) at month 5 but the buoyancy displayed no difference throughout the experimental period. The number-average molecular weight of PS pieces decreased (33% and 27% in INDG and BIOG treatment respectively), implying chain scission; accelerated (by more than 30%) sinking velocities compared to the initial weathered pieces were also measured for PS films with biofilm on their surface. The orders Rhodobacterales, Oceanospirillales and Burkholderiales dominated the distinct platisphere communities and the genera Bacillus and Pseudonocardia discriminate these assemblages from the planktonic counterpart. The functional analysis predicts overrepresentation of adhesive cells carrying xenobiotic and hydrocarbon degradation genes. Taking these into account, we can suggest that tailored marine consortia have the ability to thrive in the presence of mixtures of plastics and participate in their degradation.

Polyvinyl chloride biodegradation by Pseudomonas citronellolis and Bacillus flexus.

The accumulation of high amounts of petroleum-derived plastics in the environment has raised ecological and health concerns. The aim of this work was to study the biodegradative abilities of five bacterial strains, namely Pseudomonas chlororaphis, Pseudomonas citronellolis, Bacillus subtilis, Bacillus flexus and Chelatococcus daeguensis, towards polyethylene, polypropylene, polystyrene and polyvinyl chloride films under aerobic conditions. Preliminary screening resulted in the selection of P. citronellolis and B. flexus as potential PVC film degraders. Both strains were able to form a biofilm on the plastic film surface and to cause some modifications to the FTIR spectra of biomass-free PVC films. The two strains were then used to set up a PVC film biodegradation assay in 2-liter flasks. After 45 days incubation, fragmentation of the film was observed, suggesting that PVC biodegradative activity took place. Gel permeation chromatography analysis showed a reduction in average molecular weight of 10% for PVC incubated with P. citronellolis, with PVC polymer chains apparently attacked. Based on these results, the P. citronellolis strain was selected for biodegradation assays of two waste PVC films, used either nonsterile or subjected to ethanol sterilization. Chemical analyses on the incubated films confirmed the biodegradation of waste PVC plastics as shown by a gravimetric weight loss of up to about 19% after 30 days incubation. In summary, this work reports the biodegradation of PVC films by P. citronellolis and B. flexus. Both strains were shown to act mainly against PVC additives, exhibiting a low biodegradation rate of PVC polymer.