Monitoring of the Degradation Activities and the Diversity of the Microbial Community Degrading Refinery Waste Sludge

The biodegradation of n-alkanes and branched alkanes from waste sludge were observed in landfarming soils of Motor Oil Hellas (a petroleum refinery) and changes in the bacterial communities in the soils were monitored during the remediation. Bacterial 16S rRNA gene (rDNA)-based community fingerprint patterns were obtained from soil samples by terminal restriction fragment length polymorphism (T-RFLP) analysis. Changes in T-RFLP fingerprints, as well as in the petroleum contaminant composition of the polluted soil, correlated with degradation activities in field tests.     

Biodegradation of low-density polyethylene (LDPE) by isolated fungi in solid waste medium

In this study, biodegradation of low-density polyethylene (LDPE) by isolated landfill-source fungi was evaluated in a controlled solid waste medium. The fungi, including Aspergillus fumigatus, Aspergillus terreus and Fusarium solani, were isolated from samples taken from an aerobic aged municipal landfill in Tehran. These fungi could degrade LDPE via the formation of a biofilm in a submerged medium. In the sterilized solid waste medium, LPDE films were buried for 100 days in a 1-L flask containing 400 g sterile solid waste raw materials at 28 °C. Each fungus was added to a separate flask. The moisture content and pH of the media were maintained at the optimal levels for each fungus. Photo-oxidation (25 days under UV-irradiation) was used as a pretreatment of the LDPE samples. The progress of the process was monitored by measurement of total organic carbon (TOC), pH, temperature and moisture. The results obtained from monitoring the process using isolated fungi under sterile conditions indicate that these fungi are able to grow in solid waste medium. The results of FT-IR and SEM analyses show that A. terreus and A. fumigatus, despite the availability of other organic carbon of materials, could utilize LDPE as carbon source. While there has been much research in the field of LDPE biodegradation under solid conditions, this is the first report of degradation of LDPE by A. fumigatus.     

Biodegradability and biodegradation rate of poly(caprolactone)-starch blend and poly(butylene succinate) biodegradable polymer under aerobic and anaerobic environment

The biodegradability and the biodegradation rate of two kinds biodegradable polymers; poly(caprolactone) (PCL)-starch blend and poly(butylene succinate) (PBS), were investigated under both aerobic and anaerobic conditions. PCL-starch blend was easily degraded, with 88% biodegradability in 44 days under aerobic conditions, and showed a biodegradation rate of 0.07 day⁻¹, whereas the biodegradability of PBS was only 31% in 80 days under the same conditions, with a biodegradation rate of 0.01 day⁻¹. Anaerobic bacteria degraded well PCL-starch blend (i.e., 83% biodegradability for 139 days); however, its biodegradation rate was relatively slow (6.1 mL CH₄/g-VS day) compared to that of cellulose (13.5 mL CH₄/g-VS day), which was used as a reference material. The PBS was barely degraded under anaerobic conditions, with only 2% biodegradability in 100 days. These results were consistent with the visual changes and FE-SEM images of the two biodegradable polymers after the landfill burial test, showing that only PCL-starch blend had various sized pinholes on the surface due to attack by microorganisms. This result may be use in deciding suitable final disposal approaches of different types of biodegradable polymers in the future.     

Biodegradation of Dispersed Forties Crude and Alaskan North Slope Oils in Microcosms Under Simulated Marine Conditions

For oil spills in the open sea, operational experience has found that conventional response techniques, such as mechanical recovery, tend to remove only a small fraction of oil during major spills, a recent exception being the Mississippi River spill in Louisiana [Spill Sci. Technol. Bull. 7 (2002) 155]. By contrast, the use of dispersants can enable significant fractions of oil to be removed from the sea surface by dispersing the oil into the water column. It is thought that once dispersed the oil can biodegrade in the water column, although there is little information on the mechanism and rate of biodegradation. Two studies were undertaken on dispersion, microbial colonisation and biodegradation of Forties crude and Alaskan North Slope (ANS) oils under simulated marine conditions. The study using the Forties crude lasted 27 days and was carried out in conditions simulating estuarine and coastal conditions in waters around the UK (15 °C and in the presence of nutrients, 1 mg N-NO₃/l), while the ANS study simulated low temperature conditions typical of Prince William Sound (8 °C) and took place over 35 days. The results of both studies demonstrated microbial colonisation of oil droplets after 4 days, and the formation of neutrally buoyant clusters consisting of oil, bacteria, protozoa and nematodes. By day 16, the size of the clusters increased and they sank to the bottom of the microcosms, presumably because of a decrease in buoyancy due to oil biodegradation, however biodegradation of n-alkanes was confirmed only in the Forties study. No colonisation or biodegradation of oil was noted in the controls in which biological action was inhibited. Oil degrading bacteria proliferated in all biologically active microcosms. Without dispersant, the onset of colonisation was delayed, although microbial growth rates and population size in ANS were greater than observed with the Forties. This difference reflected the greater droplet number seen with ANS at 8 °C than with Forties crude at 15 °C. Although these studies differed by more than one variable, complicating comparison, the findings suggest that dispersion (natural or chemical) changes the impact of the oil on the marine environment, potentially having important implications for management of oil spills in relation to the policy of dispersant use in an oil spill event.     

Biodegradation Behaviors of Poly(p-dioxanone) in Different Environment Media

This work was aimed at researching the aerobic biodegradation of poly(p-dioxanone) (PPDO), a novel kind of degradable polymer material, by simulating real-life conditions in a laboratory-scale test, specified by the standard methods based on two biodegradation environments, composting and aqueous media. To measure and describe the biodegradability of PPDO, not only had carbon dioxide produced by respiratory metabolism of microorganism been measured, which determines the ultimate aerobic biodegradability of chemical compounds, but also the detailed results of biodegradation were further characterized by monitoring physical, chemical and thermal properties changes of test materials at different incubation times in the two media, confirmed by using the appropriate analytical techniques. Scanning electron microscopy was used to observe the surface morphology, and the thermal performance of PPDO was characterized by differential scanning calorimetry. The changes of molecular weight were detected by intrinsic viscosity ([η]) and gel permeation chromatography, and the variations of the molecular structure were monitored by the nuclear magnetic resonance and FT-IR. The results show that PPDO has outstanding character of biodegradation and may be more adapted for biodegrading in liquid medium than in composting.     

Biodegradation Study of Starch-graft-Acrylonitrile Copolymer

In this study the biodegradability of starch-graft-acrylonitrile (St-g-AN) copolymer has been investigated using some microorganisms including Aspergillus niger. The fungus A. niger was isolated from the soil and from the wastewater of an acrylic fiber company. The effects of four factors including environment temperature, primary inoculum concentration, pH and weight of copolymer film, on the biomass generation as a measure of biodegradation rate of copolymer, were studied using Taguchi experimental design. The statistical analysis of the results showed that the primary inoculum concentration and temperature were the most important factors affecting the biodegradation of St-g-AN copolymer. The optimum levels of temperature, pH, inoculum concentration, and weight of films to attain the maximum biodegradation (as much as 8.59 % by weight percentage during 28 days) were obtained as 30 °C, 4.75, 10⁸ spore/mL, and 1.1 g, respectively. The changes in the structure and morphological properties of the copolymer before and after degradation were determined using transform infrared spectroscopy and scanning electron microscopy.     

Use of bio‐traps with multiple substrates and electron acceptors to optimize remediation

Bio‐Traps® were used to investigate biodegradation of benzene, methyl tertiary butyl ether (MTBE), and tertiary butyl alcohol (TBA) under different conditions at a fractured rock site to aid the selection of a bioremediation approach. The Bio‐Traps were amended with the ¹³C‐labeled constituent of interest and sampled sequentially at 15‐, 30‐, 60‐, and 90‐day intervals. The conditions tested were biodegradation during operation of an air sparge system, amendment with nitrate during the air sparge operation, anaerobic biodegradation with the system turned off, and anaerobic biodegradation with nitrate amendment. There was increased biomass with nitrate amendment whether the air sparge system was on or off for all the constituents of interest. The diversity of the microbial community, determined by phospholipid fatty acid analysis, decreased with nitrate amendment as more specialized degraders were selected. The most negative indicators of potential biodegradation performance were observed with the anaerobic control. There was less biomass overall, less incorporation of ¹³C into biomass, and decreased membrane permeability. As testing with additional amendments continues at the site, it is not yet certain which treatment might be selected for bioremediation, but the Bio‐Trap tests thus far have identified that the in situ, natural attenuation condition is least favorable for biodegradation. © 2009 Wiley Periodicals, Inc.     

Characterization of Chemically Modified Potato Starch Films Through Enzymatic Degradation

The present investigation was undertaken to characterize the biodegradation pattern of chemically modified starch films. Chemically modified starch films obtained by esterification of the hydroxyl groups of the polysaccharide have shown lower water sorption than native starch films, being therefore more attractive for a number of processing applications. However, no systematic study characterizing their biodegradation behavior and comparing it with the degradation pattern of native starch films has still been published. In the current contribution we characterized the enzymatic degradation pattern of three derivatized starch films by use of a commercial α-amylase from Bacillus licheniformis. Optimum degradation conditions were chosen upon assaying the effect of enzyme load and temperature on the reaction course of native starch films. Under the conditions selected, comparison of different derivatization procedures revealed that the starch film modified with octanoyl chloride was enzymatically hydrolyzed at a much higher rate than native starch film. Maleated starch films also showed higher susceptibility to α-amylolytic hydrolysis than native starch, whereas acetylated starch showed a hydrolysis pattern similar to that of native starch. Differences in degradation rates of chemically modified films were explained in terms of their amylose content which promotes dense networks that hinder the access of starch-degrading enzymes.     

Assessment of the Whole Environmental Degradation of Oxo-Biodegradable Linear Low Density Polyethylene (LLDPE) Films Designed for Mulching Applications

The current paper is aimed at understanding the environmental fate of linear low density polyethylenes (LLDPE) films designed for mulching purposes and loaded with different pro-degradant additives. These were analyzed, upon exposure to natural sunlight for a period intended to mimick a general crop season in the mediterranean region. The selected samples underwent a relatively low extent of degradation as monitored by carbonyl index, molecular weight variation, extractability by solvent, changes in the onset of the decomposition temperature and crystallinity. The tendency to biodegradation of outdoor exposed LLDPE was then assessed under different environmental compartments including soil medium, aqueous medium as well as in axenic culture of white-rot fungus Phanerochaete chrysosporium. That fungus is known to be effective in the degradation of recalcitrant organic materials and plastic items. During the soil burial biodegradation test, lasted for 27?months, samples specimen were withdrawn at time intervals and characterized by means of structural and thermal analysis. These analytical assessments allowed to monitor any progress of oxidative degradation as a direct effect of the incubation in an active microbial environment. Analogous characterizations were carried out at the end of the biodegradation tests in aqueous medium and in P. chrysosporium axenic cultures. Data presented here are in keeping with the initial abiotic oxidation via a free radical chain reaction promoted by a pro-degradant additive acting on hydroperoxides and peroxide moieties present initially in the polymer bulk. This step was followed by a free radical cascade reactions leading to degradation once the oxidation started under relatively mild conditions (sunlight exposure). During the incubation step in soil, the abiotically degraded samples underwent significant variation in the level of oxidation and degradation with respect to the detected starting values. Indications were gained on the synergistic effect of a random fashion microbial metabolization coupled to biotically mediated oxidation of the original abiotically fragmented samples. Similar results were obtained in the biodegradation tests carried out in the aqueous media and in presence of P. chrysosporium axenic cultures. These evidences are suggesting the role of natural occurring microorganisms in promoting both partial oxiditation and degradation of LLDPE samples in combination with contextual mineralization process of the oxidized fragments.     

Assessing the biodegradation potential of polymers in screening- and long-term test systems

This paper presents a test scheme for assessing the biodegradation potential of polymers, starting with aquatic screening systems (aerobic and anaerobic) and continuing to long-term systems. At the end of the scheme the material has to prove its behavior under the relevant disposal conditions. Aerobic screening was performed mainly under aquatic conditions, but also in soil, using BOD-respirometry. Carbon balances were performed to obtain a better evaluation of the biodegradation potential. Under anaerobic conditions, biodegradation in an aquatic medium was followed by measuring CH₄ and CO₂ production. Polymers not fully degraded in the screening systems were tested in aquarium systems for at least 1 year. Biodegradation was followed by monitoring the DOC released in the water, mass loss, and microbial growth on the samples and in the water as well as via FTIR spectroscopy and SEM pictures. Results are presented for the polymers PHB, PHBV, PCL, Mater-Bi AI05H and ZF03U, and Bioceta. By combining the data from the screening with the aquarium system, a good picture of the degradation behavior of the polymers is obtained.Paper presented at the Bio/Environmentally Degradable Polymer Society—Third National Meeting, June 6–8, 1994, Boston, Massachusetts.     

A New Test Method for Determining Biodegradation of Plastic Material Under Controlled Aerobic Conditions in a Soil-Simulation Solid Environment

A new test method is described for assessing biodegradation of plastic material under simulated soil conditions. An inert substrate can be activated with soil extract and nutrient and used in place of soil in biodegradation tests. The biodegradation level is evaluated by determining the carbon dioxide (CO₂) production released by the test reactors. Effects of substrate nature, solution pH, nutrient composition, soil extract concentration, and activation duration on CO₂ production were investigated, and the experimental conditions were optimized. Results obtained with cellulose showed a biodegradation rate of 80% within 28 days. Moreover, with this kind of substrate, reaction products and residues can be easily extracted and analysed.     

Comparative Response of Indigenously Developed Bacterial Consortia on Progressive Degradation of Polyhydroxybutyrate Film Composites

The global demand of bioplastics has lead to an exponential increase in their production commercially. Hence, biodegradable nature needs to be evaluated in various ecosystems viz. air, water, soil and other environmental conditions to avoid the polymeric waste accumulation in the nature. In this paper, we investigated the progressive response of two indigenously developed bacterial consortia, i.e., consortium-I (C-I: Pseudomonas sp. strain Rb10, Pseudomonas sp. strain Rb11 and Bacillus sp. strain Rb18), and consortium-II (C-II: Lysinibacillus sp. strain Rb1, Pseudomonas sp. strain Rb13 and Pseudomonas sp. strain Rb19), against biodegradation behavior of polyhydroxybutyrate (PHB) film composites, under natural soil ecosystem (in net house). The biodegraded films recovered after 6 and 9 months of incubation were analyzed through Fourier transform infrared spectroscopy and scanning electron microscopy to determine the variations in chemical and morphological parameters (before and after incubation). Noticeable changes in the bond intensity, surface morphology and conductivity were found when PHB composites were treated with C-II. These changes were drastic in case of blends in comparison to copolymer. The potential isolates not only survived, but, also, there was a significant increase in bacterial diversity during whole period of incubation. To the best of our knowledge, it is the first report which described the biodegradation potential of Lysinibacillus sp. as a part of C-II with Pseudomonas sp. against PHB film composites.     

Composting kinetics in full-scale mechanical–biological treatment plants

This study focuses on the investigation of the kinetics of municipal solid waste composting in three full-scale mechanical–biological treatment (MBT) plants. The aims were to test a kinetic model based on volatile solids (VS) content change for describing the composting process in MBT plants, and to identify the model parameters that affected the estimation of the reaction rate constant most. To achieve this, VS content and several environmental conditions, namely temperature, moisture content, oxygen concentration and total bulk density were monitored throughout the composting process. Experimental data was fitted with a first-order kinetic model, and a rate constant (k) characteristic of composting under optimum environmental conditions was obtained. The kinetic model satisfactorily described the experimental data for the three MBT plants. k values ranged from 0.043 ± 0.002 d^?1 to 0.082 ± 0.011 d^?1. Sensitivity analysis showed that the model parameters that most affected the estimation of k were the initial biodegradable volatile solids content, the maximum temperature for biodegradation and the optimum moisture content. In conclusion, we show for the first time that full-scale MBT plants can be successfully modelled with a composting kinetic model.     

Assessing Aerobic Biodegradability of Plastics in Aqueous Environment by GC-Analyzing Composition of Equilibrium Gaseous Phase

Testing biodegradability of plastics under varied conditions of the environment as well as under laboratory conditions in accordance with valid international standards is very laborious, lengthy and often also economically demanding. For this reason, applicability was verified of gas chromatography to analyze gaseous phase when investigating the biodegradation course of plastics in an aqueous environment as an alternative to customary employed methods. A mathematical model of acid–basic CO₂ equilibrium in a gas–liquid system was worked out, enabling to determine quantity of produced CO₂ through chromatographic analysis of gaseous phase, in dependence on ratio of liquid and gas phase volumes (V _l/V _g) and on actual pH of liquid phase. Experimental conditions for organizing the tests were optimized. A ratio that proved suitable was V _l/V _g ≅ 0.1 at pH ≈ 7.1 of liquid phase. Under these test conditions, biodegradability of model samples, PHB, Gellan gum and Xanthan gum, was explored; course of biodegradation was studied through produced CO₂ (values ) determined by analyzing gaseous phase through gas chromatography on the one hand, and through customary “titration” procedure on the other. With water-soluble polymers, the decrement in dissolved organic carbon (values D _DOC) was also studied. Difference between values does not exceed 5%. The procedures in question are alternative “substituting” procedures for observing course of aerobic biodegradation of substances in an aqueous environment.     

Assessment of internal condition of waste in a roofed landfill

Recently, roofed landfills have been gaining popularity in Japan. Roofed landfills have several advantages over non-roofed landfills such as eliminating the visibility of waste and reducing the spread of offensive odours. This study examined the moisture balance and aeration conditions, which promote waste stabilisation, in a roofed landfill that included organic waste such as food waste. Moisture balance was estimated using waste characterization and the total amount of landfilled waste. Internal conditions were estimated based on the composition, flux, and temperature of the landfill gas. Finally, in situ aeration was performed to determine the integrity of the semi-aerobic structure of the landfill.With the effects of rainfall excluded, only 15% of the moisture held by the waste was discharged as leachate. The majority of the moisture remained in the waste layer, but was less than the optimal moisture level for biodegradation, indicating that an appropriate water spray should be administered. To assess waste degradation in this semi-aerobic landfill, the concentration and flow rate of landfill gas were measured and an in situ aeration test was performed. The results revealed that aerobic biodegradation had not occurred because of the unsatisfactory design and operation of the landfill.     

Volatilization and Biodegradation of VOCs in Membrane Bioreactors (MBR)

Volatilization and biodegradation are major competitive volatile organic compound (VOC) removal mechanisms in biological wastewater treatment process, which depend on compound specific properties and system design/operational parameters. In this study, a mathematical model was used to determine major removal pathways at various organic loading rates (OLR), solids residence time (SRT) and dissolved oxygen (DO) concentrations in a biological process for vinyl acetate. Model results showed that biological treatment process should be designed with long SRT, high OLR and low DO concentrations to maximize biodegradation and minimize volatilization of VOCs. Unless a VOC is toxic to microorganisms under the given conditions, low VOC emission rates are an inherent advantage of MBRs, which operate at higher OLR and longer SRT compared to conventional activated sludge process. A lab scale membrane bioreactor (MBR) was operated at varying OLR to investigate the relative volatilization and biodegradation rates for acetaldehyde, butyraldehyde and vinyl acetate. Synthetic wastewater containing three VOCs was introduced to the MBR. The DO concentration and SRT was maintained at 2.0 mg L^{− 1} and 100 days, respectively. The overall VOC removal rate was more than 99.7% for three VOCs at all the OLR. For vinyl acetate, the biodegradation rate increased from 93.87 to 99.40% and the volatilization removal rate decreased from 6.09 to 0.59% as OLR was increased from 1.1 to 2.0 kg COD m^{− 3} d^{− 1}. It was confirmed that a MBR can be a promising solution to reduce VOC emissions from wastewater.     

Effects of physicochemical factors on PAH degradation by Planomicrobium alkanoclasticum

Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants that are mutagenic, carcinogenic, and toxic to living organisms. Here, the ability and effectiveness of selected bacteria isolated from an oil‐contaminated area in biodegrading PAHs were evaluated, and the optimal conditions conducive to bacterial PAH biodegradation were determined. Of six bacterial isolates identified based on their 16S rRNA sequences, Planomicrobium alkanoclasticum could subsist on and consume nearly all hydrocarbons according to the 2,6‐dichlorophenolindophenol assay. The efficacy of this isolate at PAH biodegradation was then empirically confirmed. After 30 days of incubation, P. alkanoclasticum degraded 90.8% of the 16 PAH compounds analyzed and fully degraded eight of them. The optimum P. alkanoclasticum growth conditions were 35°C, pH 7.5, and NaNO₃ as the nitrogen source. Under these biostimulant conditions, P. alkanoclasticum degraded 91.4% of the total PAH concentration and completely decomposed seven PAHs after 15 days incubation. Hence, P. alkanoclasticum is an apt candidate for the biodegradation of PAHs and the bioremediation of sites contaminated by them.     

Influence of pH and C/N Ratio on Poly(Vinyl Alcohol) Biodegradation in Mixed Bacterial Culture

Cultivation conditions affecting poly(vinyl alcohol) (PVA) degradation by a mixed bacterial culture of Bacillus sp. and Curtobacterium sp. were investigated. Bacterial strains used in this study were isolated from the watercourse and the sewage sludge of vinylonfibre mill by enrichments on PVA as the sole carbon source. The results showed that PVA was greatly degraded under the following conditions: 0.5% PVA as a substrate at the initial medium pH of 8 with 0.15% glucose and urea at C/U ratio 1.5:1 and 1% bacterial inoculum, at a temperature of 35 °C and a shaking speed of 110 rpm. The analysis of FTIR and ¹H NMR spectra before and after biodegradation indicate fission of the PVA molecular chain during the incubation.     

Volatile Organic Compounds Selection for Incorporation in Photochemical Mechanisms and the Development of Secondary Pollution Reduction Strategies

In the present work a method for the evaluation of the importance of the VOCs species is presented, aiming to provide criteria for the incorporation of these species into atmospheric photochemical mechanisms and for the successful application of secondary pollution reduction strategies. According to the method presented here, the species can be divided into more important and less important ones, taking into account their mixing ratios and emission values in combination with their reactivity. For this classification three quantitative and one qualitative criteria were introduced. Overall, it is concluded that alkenes with more than a few carbon atoms in their chain appear to be more important in urban and suburban areas, while in background conditions the alkanes, having the smaller chain (ethane, propane), become more important. In the case of alkenes there is no clear species classification, except for the biogenically emitted compounds, isoprene and limonene. In general, more important alkenes appear to be those with the smaller chain (ethene, propene, butene). Most abundant aromatics are benzene, toluene, and xylene. In background conditions higher aromatics are also important, especially 1,2,3-, 1,3,5-, and 1,2,4-trimethylbenzene. The most important carbonylic compounds are formaldehyde, acetaldehyde, and acetone. Finally, taking into account the results mentioned above, a new photochemical mechanism was developed. The species and species groups used in the proposed mechanism are: ethane, higher alkanes, ethene, propene, 2- butene, 1-alkenes, 2-alkenes, higher alkenes, benzene, toluene, m-, o-, p-xylene, 1,3,5-, 1,2,3-, 1,2,4-trimethylbenzene, higher aromatics, formaldehyde, acetaldehyde, higher aldehydes, isoprene, limonene, and other biogenic VOCs.