Microorganisms that produce enzymes active on biodegradable polyesters are ubiquitous.

Biodegradability standards measure ultimate biodegradation of polymers by exposing the material under test to a natural microbial inoculum. Available tests developed by the International Organization for Standardization (ISO) use inoculums sampled from different environments e.g. soil, marine sediments, seawater. Understanding whether each inoculum is to be considered as microbially unique or not can be relevant for the interpretation of tests results. In this review, we address this question by consideration of the following: (i) the chemical nature of biodegradable plastics (virtually all biodegradable plastics are polyesters) (ii) the diffusion of ester bonds in nature both in simple molecules and in polymers (ubiquitous); (iii) the diffusion of decomposers capable of producing enzymes, called esterases, which accelerate the hydrolysis of esters, including polyesters (ubiquitous); (iv) the evidence showing that synthetic polyesters can be depolymerized by esterases (large and growing); (v) the evidence showing that these esterases are ubiquitous (growing and confirmed by bioinformatics studies). By combining the relevant available facts it can be concluded that if a certain polyester shows ultimate biodegradation when exposed to a natural inoculum, it can be considered biodegradable and need not be retested using other inoculums. Obviously, if the polymer does not show ultimate biodegradation it must be considered recalcitrant, until proven otherwise.

Biodegradation of plastics in soil and effects on nitrification activity. A laboratory approach.

The progressive application of new biodegradable plastics in agriculture calls for improved testing approaches to assure their environmental safety. Full biodegradation (≥90%) prevents accumulation in soil, which is the first tier of testing. The application of specific ecotoxicity tests is the second tier of testing needed to show safety for the soil ecosystem. Soil microbial nitrification is widely used as a bioindicator for evaluating the impact of chemicals on soil but it is not applied for evaluating the impact of biodegradable plastics. In this work the International Standard test for biodegradation of plastics in soil (ISO 17556, 2012) was applied both to measure biodegradation and to prepare soil samples needed for a subsequent nitrification test based on another International Standard (ISO 14238, 2012). The plastic mulch film tested in this work showed full biodegradability and no inhibition of the nitrification potential of the soil in comparison with the controls. The laboratory approach suggested in this Technology Report enables (i) to follow the course of biodegradation, (ii) a strict control of variables and environmental conditions, (iii) the application of very high concentrations of test material (to maximize the possible effects). This testing approach could be taken into consideration in improved testing schemes aimed at defining the biodegradability of plastics in soil.

Monitoring biodegradation of poly(butylene sebacate) by Gel Permeation Chromatography, (1)H-NMR and (31)P-NMR techniques.

The increasing use of new generation plastics has been accompanied by the development of standard methods for studying their biodegradability. Generally, test methods are based on the measurement of CO(2) production, i.e. the mineralization degree of the tested materials. However, in order to describe the biodegradation process, the determination of the residual amount of tested material which remains in the environment and its chemical characterization can be very important. In this study, the biodegradation in soil of a model polyester (poly(butylene sebacate)) was monitored. Gel Permeation Chromatography and Nuclear Magnetic Resonance ((31)P-NMR and (1)H-NMR) were used in order to obtain information about the polyester structure and the possible by-products that can be found in soil during and at the end of the incubation. The polyester mineralization (i.e. the CO(2) production) was tested according to ASTM 5988 standard method for 245 days. When the polyester mineralization was about 21% and 37% (after 78 and 140 days of incubation) and at the end of the process (63% of mineralization, 100% if compared to the cellulose used as reference material), the soil was extracted with chloroform (solvent of the tested substance) and the extracts were analyzed using GPC and NMR acquisitions. The analytical acquisitions showed high molecular weight polyester in soil during the incubation (78 and 140 days): the polyester concentration decreased but its structure remained almost the same with a slow decreasing in molecular weight. At the end of the test (245 days) no film of the polyester could be extracted from the soil: NMR acquisitions and GPC analyses of the extracts suggested a strong degraded structure of the residual polyester. Even if at the end of the process only 63% of carbon had been lost by mineralization, the whole of the added polyester seems to have disappeared after about eight months of incubation, suggesting substantial biomass formation.

Laboratory test methods to determine the degradation of plastics in marine environmental conditions.

In this technology report, three test methods were developed to characterize the degradation of plastic in marine environment. The aim was to outline a test methodology to measure the physical and biological degradation in different habitats where plastic waste can deposit when littered in the sea. Previously, research has focused mainly on the conditions encountered by plastic items when floating in the sea water (pelagic domain). However, this is just one of the possible habitats that plastic waste can be exposed to. Waves and tides tend to wash up plastic waste on the shoreline, which is also a relevant habitat to be studied. Therefore, the degradation of plastic items buried under sand kept wet with sea water has been followed by verifying the disintegration (visual disappearing) as a simulation of the tidal zone. Most biodegradable plastics have higher densities than water and also as a consequence of fouling, they tend to sink and lay on the sea floor. Therefore, the fate of plastic items lying on the sediment has been followed by monitoring the oxygen consumption (biodegradation). Also the effect of a prolonged exposure to the sea water, to simulate the pelagic domain, has been tested by measuring the decay of mechanical properties. The test material (Mater-Bi) was shown to degrade (total disintegration achieved in less than 9 months) when buried in wet sand (simulation test of the tidal zone), to lose mechanical properties but still maintain integrity (tensile strength at break = -66% in 2 years) when exposed to sea water in an aquarium (simulation of pelagic domain), and substantially biodegrade (69% in 236 days; biodegradation relative to paper: 88%) when located at the sediment/sea water interface (simulation of benthic domain). This study is not conclusive as the methodological approach must be completed by also determining degradation occurring in the supralittoral zone, on the deep sea floor, and in the anoxic sediment.

Kinetics of monomer biodegradation in soil.

In modern intensive agriculture, plastics are used in several applications (i.e. mulch films, drip irrigation tubes, string, clips, pots, etc.). Interest towards applying biodegradable plastics to replace the conventional plastics is promising. Ten monomers, which can be applied in the synthesis of potentially biodegradable polyesters, were tested according to ASTM 5988-96 (standard respirometric test to evaluate aerobic biodegradation in soil by measuring the carbon dioxide evolution): adipic acid, azelaic acid, 1,4-butanediol, 1,2-ethanediol, 1,6-hexanediol, lactic acid, glucose, sebacic acid, succinic acid and terephthalic acid. Eight replicates were carried out for each monomer for 27-45 days. The numerical code AQUASIM was applied to process the CO2 experimental data in order to estimate values for the parameters describing the different mechanisms occurring to the monomers in soil: i) the first order solubilization kinetic constant, K(sol) (d⁻¹); ii) the first order biodegradation kinetic constant, K(b) (d⁻¹); iii) the lag time in biodegradation, t(lag) (d); and iv) the carbon fraction biodegraded but not transformed into CO2, Y (-). The following range of values were obtained: [0.006 d⁻¹, 6.9 d⁻¹] for K(sol), [0.1 d⁻¹, 1.2 d⁻¹] for K(b), and [0.32-0.58] for Y; t(lag) was observed for azelaic acid, 1,2-ethanediol, and terephthalic acid, with estimated values between 3.0 e 4.9 d.