Biodegradation of Whey Protein-Based Edible Films

The biodegradability of the edible films made of whey proteins by disulfide cross-linking was investigated. Whey protein concentrate (WPC) and whey protein isolate (WPI) films were subjected to microbial degradation using Pseudomonas aeruginosa and composting burial degradation. Results from the microbial degradation showed that whey protein films could support the growth of P. aeruginosa. The bacterial growth characteristics were well described using the Gompertz model. WPC films degraded faster than WPI films, suggesting that the biodegradability of protein films is associated with the film composition and the extent of covalent cross-linking. WPI films buried in a compost pile began to degrade in two days and became darker over time. More than 80% of total solids were lost in 7 days.     

Bacterial degradation of a new polyester,polyethylene glycol-phthalate polyester

Microorganisms which can assimilate a new polyester synthesized from polyethylene glycol (PEG) as a dihydroxyl compound and phthalic acid as a dicarboxyl compound were isolated from soils by enrichment culture techniques. Two cultures, K and N, were obtained: Culture K grew on PEG 4000 polyester and culture N assimilated PEG 6000 polyester. Each culture included two bacteria indispensable for the degradation of polyesters: bacteria K1 and K2 for PEG 4000 polyester-utilizing culture K and bacteria N1 and N2 for PEG 6000 polyester-utilizing culture N. Bacteria K2 and N2 were responsible for the hydrolysis of ester bonds in a polyester and both were identified as the same species,Comamonas acidovorans. Bacteria K1 and N6 could assimilate PEG as a sole carbon and energy source. Both are Gram-negative, non-spore-forming rods and resembled each other on their colony characteristics, although strain K1 could not grow on PEG 6000.C. acidovorans N2 (K2) grew on dialkyl phthalates (C₂–C₄) and phthalate and tributyrin, but not on PEG, diphthalic PEG, and PEG phthalate polyesters. Their culture supernatant and washed cells hydrolyzed PEG (400–20,000) phthalate and sebacate polyesters.C. acidovorans had higher esterase activity toward PEG phthalate, isophthalate, and terephthalate polyesters than known esterase and lipases. The esterase seemed to be an extracellular one and attached to the cell surface.     

Microbial degradation of poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in compost

The microbial degradation of tensile test pieces made of poly(3-hydroxybutyrate) [P(3HB)] or copolymers with 10% [P(3HB-co-10%3HV)] and 20% [P(3HB-co-20%3HV)] 3-hydroxyvaleric acid was studied in small household compost heaps. Degradation was measured through loss of weight (surface erosion) and changes in molecular weight and mechanical strength. It was concluded, on the basis of weight loss and loss of mechanical properties, that P(3HB) and P(3HB-co-3HV) plastics were degraded in compost by the action of microorganisms. No decrease inM _w could be detected during the degradation process. The P(3HB-co-20%3HV) copolymer was degraded much faster than the homopolymer and P(3HB-co-10%3HV). One hundred nine microbial strains capable of degrading the polymersin vitro were isolated from the samples used in the biodegradation studies, as well as from two other composts, and identified. They consisted of 61 Gram-negative bacteria (e.g.,Acidovorax facilis), 10 Gram-positive bacteria (mainlyBacillus megaterium), 35Streptomyces strains, and 3 molds.     

Assessment of Several Test Methods for the Determination of the Anaerobic Biodegradability of Polymers

Anaerobic degradation of eight commercially available biodegradable polymers was compared in two anaerobic tests using digestion sludge, according to ISO 11734 and ASTM D.5210-91. Cotton, polyhydroxybutyrate/hydroxyvalerate copolymer (PHB/PHV), starch blend, thermoplastic cellulose acetate, and cellulose acetate fibers proved to be anaerobically degradable, but only a low extent of degradation was found for polylactide, polyvinylalcohol, and polycaprolactone. Both test methods gave the same overall results, but with the ISO medium, longer lag phases and greater ranges of variation in the results were observed. These effects are presumably due to low concentrations of carbon dioxide in the ISO medium. Carbon dioxide has been demonstrated to be essential for the growth of various anaerobic bacteria, notably homoacetogenic and methanogenic bacteria.     

Characterization of Aerobic Bacteria Involved in Degrading Polyethylene Glycol (PEG)-3400 Obtained by Plating and Enrichment Culture Techniques

Polyethylene glycol (PEG) 3400-degrading aerobic bacteria were isolated from tap water and wetland sediments and then characterized. Only one Sphingomonas strain was obtained in enrichment cultures from each inoculum source whereas a total of 15 bacterial strains were isolated on agar plates. Nine of the 15 isolates were confirmed as PEG 3400 degraders. Three of the 9 PEG 3400 degraders were Gram-negative bacteria belonging to the genus Pseudomonas and genus Sphingomonas. The remaining six isolates were Gram-positive bacteria belonging to genera Rhodococcus, Williamsia, Mycobacterium and Bacillus. PEG 3400 was quantified at 194 nm spectrophotometrically and, at the same time, the growth of two Gram-negative (isolates P1 and P7) and five Gram-positive (isolates P2, P3, P4, P5 and P6) PEG 3400-degrading bacteria were assayed in liquid media and on agar plates amended with PEG 3400, and also on Nutrient Agar plates and pure agar plates without PEG 3400 addition. No growth was observed on the pure agar plates for all the tested strains for a period of 31 days. All tested PEG 3400 degraders showed much lower viability in liquid culture than on the corresponding agar plates in the presence of PEG 3400. Two Gram-negative isolates P1 and P7 did not show significant growth advantage over the Gram-positive isolates both on the agar plates and in the liquid medium amended with PEG 3400. Our results suggest that diversity of PEG degrading bacteria is high in the environments and culturing techniques affect the successful isolation of the bacteria responsible for degradation.     

Enhancing Performance of Sphingobacterium spiritivorum in Bioremediation Phenanthrene Contaminated Sand

Many biodegradation studies have focused on survival of isolated bacteria to increase the bacteria population and subsequently enhance the efficiency of polycyclic aromatic hydrocarbon (PAH) biodegradation. However, there is limited research on enhancing the performance of isolated bacteria through reinoculation. Thus, this study was designed to evaluate the effects of reinoculation on the performance of Sphingobacterium spiritivorum in degradation of phenanthrene contaminated sand. Experiments were performed in three different reactors. Inoculation was performed once (day 0) in reactor 1. In reactor 2, inoculation was performed twice (day 0 and day 5). The bacteria was isolated from reactor 2 and inoculated into reactor 3. The study results show reactor 3 having the highest degradation rate (13.61 mg/kg/day) and percentage removal (95.36 percent). In contrast, without reinoculation in reactor 1, 68.93 percent of phenanthrene was removed. Thus, the performance of S. spiritivorum in phenanthrene degradation can be enhanced through reinoculation. © 2014 Wiley Periodicals, Inc.     

Improvement of digestibility of sunflower and corn residues by some saprophytic fungi

The in vitro degradation of fiber from two agricultural residues (Helianthus annuus and Zea mays) was studied using four saprophytic fungi. The changes in cellulolytic enzyme and phosphatase activities, the C/N ratio, the dry weight, the crude fiber (CF) content, and the protein and fiber digestibility coefficients were investigated in solid-state culture during a 45-day period of biodegradation. The FPase activity of Phanerochaete chrysosporium and the endoglucanase activities of P. chrysosporium and Trichoderma reesei compared to those of the other fungi were significantly (P < 0.05) high on both crop resides. Both acid and alkaline phosphatases were produced in greater amounts by P. chrysosporium than by the other fungi. The organic carbon contents of both residues were reduced more by T. reesei than by the other fungi. The fiber digestibility coefficient of untreated corn residue was higher than that of sunflower residue, but after treatment with fungi, the sunflower fiber digestibility increased more than that of corn did. Although the CF contents of both residues were decreased more by P. chrysosporium than by T. reesei, the fiber digestibility coefficients of both residues were increased more by T. reesei than by P. chrysosporium. The treatment of both crop residues increased their protein digestibility coefficients significantly compared to the control. The protein digestibility coefficients of both residues were increased more by T. reesei and P. chrysosporium than by the other fungi. In all treatments, the protein digestibility coefficient of sunflower residue was higher than that of corn residue.     

Biodegradation of chlorinated organic pesticides endosulfan and chlorpyrifos in soil extract broth using fungi

The present study was designed to screen 20 fungi for their potential to degrade the chlorinated organic pesticides endosulfan and chlorpyrifos. Fungi were first screened for their tolerance to various concentrations of target pesticides using soil extract agar and subsequent degradation studies were performed in soil extract broth containing 25 mg/L of the individual pesticide. Pesticide degradation was evaluated using gas chromatography. Other parameters, such as pH and mycelial weight, were also determined. Based on percent growth inhibition of test fungi and subsequent analysis of EC₅₀ values, the overall results revealed that chlorpyrifos showed significantly more growth inhibition in all tested fungi compared with endosulfan. Trametes hirsuta showed complete degradation of both α‐ and β‐endosulfan isomers and Cladosporium cladosporioides displayed maximum degradation of chlorpyrifos. All test fungi degraded endosulfan more efficiently than chlorpyrifos, except Phanerochaete chrysosporium, Trichoderma harzianum, and Trichoderma virens which showed higher degradation of chlorpyrifos than endosulfan. It was also found that all tested fungi degraded α‐endosulfan more efficiently than β‐endosulfan. Endosulfan sulfate was found to be the major degradation product with all tested fungi. Fungi which showed more endosulfan degradation also produced more endosulfan sulfate. However, less endosulfan sulfate was detected with T. hirsuta and Trametes versicolor, although they degraded endosulfan more efficiently.     

Influence of Processing Additives on the Degradation of Melt-Pressed Films of Poly(ε-Caprolactone) and Poly(Lactic Acid)

Melt-pressed films of polycaprolactone (PCL) and poly(lactic acid) (PLA) with processing additives, CaCO₃, SiO₂, and erucamide, were subjected to pure fungal cultures Aspergillus fumigatus and Penicillium simplicissimum and to composting. The PCL films showed a rapid weight loss with a minor reduction in the molecular weight after 45 days in A. fumigatus. The addition of SiO₂ to PCL increased the rate of (bio)erosion in A. fumigatus and in compost. The use of a slip additive, erucamide, was shown to modify the properties of the film surface without decreasing the rate of bio(erosion). Both the rate of weight loss and the rate of molecular weight reduction of PCL increased with decreasing film thickness. The addition of CaCO₃ to PLA significantly reduced the thermal degradation during processing, but it also reduced the rate of the subsequent (bio)degradation in the pure fungal cultures. PLA without additives and PLA containing SiO₂ exhibited the fastest (bio)degradation, followed by PLA with CaCO₃. The degradation of the PLA films was initially governed by chemical hydrolysis, followed by an acceleration of the weight change and of the molecular weight reduction. PLA film subjected to composting exhibits a rapid decrease in molecular weight, which then remains unchanged during the measurement period, probably because of crystallization.     

Pure-culture and enzymatic assay for starch-polyethylene degradable plastic biodegradation withStreptomyces species

Eleven starch-polyethylene degradable plastic films were prepared from masterbatches from Archer Daniels Midland Inc. (ADM), EcoStar Inc. (SLS), and Fully Compounded Plastic Inc. The biodegradability of initial and 70°C heat-treated materials was determined using a pure-culture assay withStreptomyces badius 252,S. setonii 75Vi2, orS. viridosporus T7A or without bacterial culture (control). Films were treated with 10-foldS. setonii culture concentrates and compared with inactive enzyme controls. Changes in each films mechanical property, molecular weight distribution, and Fourier-transformed infrared spectrum (FT-IR) were determined, and results were evaluated for significant differences by analysis of variance. Cell mass accumulation on each film was quite pronounced. In pure-culture studies, biodegradation was demonstrated for ADM-7 and SLS-2 initial films and for ADM-6 heat-treated films, whereas after 3-week treatment with activeS. setonii culture concentrates (enzyme assay), reductions in mechanical properties and changes in FT-IR spectrum were illustrated by all the films except SLS-2. Thus the absence of biofilm formation on the film surface permitted enzymatic attack of the materials. Furthermore, inhibition of chemical oxidative degradation in the pure-culture assay was demonstrated for ADM-11, SLS-5, and SLS-10 initial materials and for ADM-4, ADM-7, SLS-8, and SLS-10 heat-treated films. These data suggest that biological and chemical degradation were directly affected by the reduction in oxygen tension on the plastic film surface due to cell mass accumulation. This same phenomenon could be the cause for slow degradation rates in nature.Journal Paper No. J-15061 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Project Nos. 0178 and 2889.     

Stem Growth of Picea Abies in South Western Sweden in the 10 Years Following Liming and Addition of PK and N

Liming and/or application of specific nutrients have been proposed as countermeasures to the acidification of forest soils in southern Sweden. In this study the stem growth of Picea abies (L.) Karst. growing on acidic mineral soils in SW Sweden was investigated 10 years after additions of lime (Ca; 3000 kg lime ha⁻¹), lime plus P (25 kg ha⁻¹) and K (80 kg ha⁻¹), or N in low doses (2 × 10 kg ha⁻¹ yr⁻¹) (treatments: CaPK, Ca, N, CaPKN, and 2Ca2P2K, respectively). Compared with the control, stem growth was increased following all treatments involving lime additions, including liming alone. The PK addition did not seem to affect growth. The most plausible cause of the observed growth increases was that the lime additions indirectly increased the supply of plant-available N. The annual low-dose N addition did not significantly affect growth. This suggests that air-borne deposition of N, which supplies very small doses of N throughout the year, has a minor or even negligible influence on P. abies growth.     

Inactivation of pathogenic micro-organisms in hospital waste using a microwave

The aim of this study was to investigate the efficiency of dry sterilization by microwave in the inactivation of pathogenic bacteria in blood wastes. Eight bacteria strains were selected for this purpose and these are S. aureus, Salmonella sp., E. faecalis, K. pneumonia, P. aeruginosa, E. coli, Streptococcus Group B and Bacillus sp. Sterilization by microwave was carried out at 385, 450 and 700 watts for 1, 5 and 10 min, respectively. Isolation and enumeration of pathogenic bacteria before and after treatments were carried out using direct plate technique and inactivation was determined in terms of log reduction. Increasing the running time and power to 10 min and 700 watts did not increase log reduction of most of pathogenic bacteria especially of gram negative bacteria. Revival of pathogenic bacteria during the storage period might be aided by the presence of a low moisture content which helps bacterial cells to revive but not to reproduce or over-grow. It can be concluded that dry sterilization of blood wastes by microwave reduces the number and limited re-growth of these pathogenic bacteria during storage time, due to the lower moisture content in the blood wastes.     

Anammox: an option for ammonium removal in bioreactor landfills

Experiments carried out in bioreactor landfill simulators demonstrated that more than 40% of the total N was transferred into the liquid and gas phases during the incubation period of 380 days. Ammonium, an end product of protein degradation and important parameter to consider during landfill closure, tends to accumulate up to inhibitory levels in the leachate of landfills especially in landfills with leachate recirculation. Most efforts to remove ammonium from leachate have been focused on ex situ and partial in situ methods such as nitrification, denitrification and chemical precipitation. Besides minimal contributions from other N-removal processes, Anammox (Anaerobic Ammonium Oxidation) bacteria were found to be active within the simulators. Anammox is considered to be an important contributor to remove N from the solid matrix. However, it was unclear how the necessary nitrite for Anammox metabolism was produced. Moreover, little is known about the nature of residual nitrogen in the waste mass and possible mechanisms to remove it. Intrusion of small quantities of O₂ is not only beneficial for the degradation process of municipal solid waste (MSW) in bioreactor landfills but also significant for the development of the Anammox bacteria that contributed to the removal of ammonium. Volatilisation and Anammox activity were the main N removal mechanisms in these pilot-scale simulators. The results of these experiments bring new insights on the behaviour, evolution and fate of nitrogen from solid waste and present the first evidence of the existence of Anammox activity in bioreactor landfill simulators.     

Biodegradation of pretreated polyethylene film by Pseudomonas aeruginosa AMB-CD-1

A comparative study evaluated the acid, alkali, and heat-treated polyethylene biodegradation efficiency of Pseudomonas aeruginosa AMB-CD-1. The polyethylene (PE) pieces were separately treated with heat (50°C), acid (1N HCl), and alkali (1N NaOH) and then washed with water before use. All the treated samples were analyzed through thermogravimetric analysis. In addition, weight and temperature changes during the decomposition reactions were also measured and determined. In these treatments, the PE films of heat-treated and acid-treated low-density polyethylene (LDPE) indicated more significant weight loss at 120°C (48.99% and 40.75%, respectively) as compared to their control or untreated PE and alkali-treated LDPE (21.84% and 24.68%, respectively). A biodegradation assay was then conducted with treated and untreated LDPE films with P. aeruginosa AMB-CD-1 strain. Fourier transform infrared spectroscopy analysis revealed that the heat or acid-pretreated samples with isolate AMB-CD-1 displayed peaks at 2922.84, 2923.97, and 1450.31, 874.22 cm⁻¹ for C–H stretching deformation vibration, CH₂ scissoring vibration, –CHO stretching, and strong alkyl structure, respectively. Furthermore, the new peaks with a significant difference at 2500–2000 cm⁻¹ (O═C═O, O–H stretching vibration: carboxylic acid) and 1500–1000 cm⁻¹ (–CHO and C═O stretching) were noticed in the infrared spectral range of LDPE degradation. Modifications in the functional group provided evidence that biodegradation had impacted the chemical structure of the LDPE film. Additionally, it was demonstrated that pretreating LDPE films with heat or acid could speed up their biodegradation.     

Degradation of Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Some Soil Aspergillus spp.

Systematic screening of 45 soil fungi for degradation polyhydroxyalkanoic acids (PHAs) has led to the selection of 6 potent Aspergillus isolates belonging to A. flavus, A. oryzae, A. parasiticus, and A. racemosus. Degradation of PHAs as determined by tube assay method revealed that these Aspergillus spp. were more efficient in degrading poly(3-hydroxybutyrate) [P(3HB)] compared to copolymer of 3-hydroxybutyric acid and 3-hydroxyvaleric acid (P3HB-co-16% 3HV). Moreover, the extent of degradation in mineral base medium was much better than those in complex organic medium. For all the Aspergillus spp. tested, maximum degradation was recorded at a temperature of 37°C with significant inhibition of growth. The optimum pH range for degradation was 6.5–7.0 with degradation being maximum at pH 6.8. The extent of polymer degradation increased with increase in substrate concentration, the optimum concentration for most of the cultures being 0.4% and 0.2% (w/v) for P(3HB) and P(3HB-co-16%3HV) respectively. Supplementation of the degradation medium with additional carbon sources exerted significant inhibitory effect on both P(3HB) and P(3HB-co-16%3HV) degradation.     

Early Effects of Atmospheric Ammonia Deposition on Calluna Vulgaris (L.) Hull Growing on an Ombrotrophic Peat Bog

This paper reports data from a field study investigating the impacts of elevated ammonia (NH₃) deposition on Calluna vulgaris growing on an ombrotrophic peat bog in S.E. Scotland. Shoot extension, foliar N concentrations, chlorophyll concentration and chlorophyll fluorescence were measured during the second growing season of exposure to a gradient of ammonia concentrations. Results indicate that NH₃ increases growth between 150–200 kg N ha^–1y^–1 cumulative deposition. Foliar N content increased significantly in response to NH₃ cumulative deposition up to 400 kg N ha^–1 y^–1 whereas chlorophyll a content significantly decreased. Measurements of Fv/Fm suggest that although NH₃ exposure altered the growth and reduced chlorophyll a, the efficiency of photosystem II was insensitive to NH₃-N deposition at this stage.     

Biodegradation of Epoxy and MF Modified Polyurethane Films Derived from a Sustainable Resource

Mesua ferrea L. seed oil (MFLSO) modified polyurethanes blends with epoxy and melamine formaldehyde (MF) resins have been studied for biodegradation with two techniques, namely microbial degradation (broth culture technique) and natural soil burial degradation. In the former technique, rate of increase in bacterial growth in polymer matrix was monitored for 12 days via a visible spectrophotometer at the wavelength of 600 nm using McFarland turbidity as the standard. The soil burial method was performed using three different soils under ambient conditions over a period of 6 months to correlate with natural degradation. Microorganism attack after the soil burial biodegradation of 180 days was realized by the measurement of loss of weight and mechanical properties. Biodegradation of the films was also evidenced by SEM, TGA and FTIR spectroscopic studies. The loss in intensity of the bands at ca. 1735 cm⁻¹ and ca. 1050 cm⁻¹ for ester linkages indicates biodegradation of the blends through degradation of ester group. Both microbial and soil burial studies showed polyurethane/epoxy blends to be more biodegradable than polyurethane/MF blends. Further almost one step degradation in TG analysis suggests degradation for both the blends to occur by breakage of ester links. The biodegradation of the blends were further confirmed by SEM analyses. The study reveals that the modified MFLSO based polyurethane blends deserve the potential to be applicable as “green binders” for polymer composite and surface coating applications.     

Enzymatic Degradation and Aminolysis of Microbial Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) Single Crystals

Solution-grown single crystals of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] were hydrolyzed by polyhydroxybutyrate (PHB) depolymerase from Ralstonia pickettii T1. Enzymatic degradation proceeded from the edges of lamellar crystals, yielding serrated contour and small crystal fragments. Gel permeation chromatography analysis revealed that the molecular weights of the crystals decreased during enzymatic degradation, suggesting that the enzymatic hydrolysis of chain-folding regions at the crystal surfaces occurred in addition to the enzymatic degradation at crystal laterals or edges. After P(3HB-co-4HB) single crystals were aminolysed in 20% aqueous methylamine solution to remove the folded-chain regions and enzymatic degradation by lipase from Rhizopus oryzae to remove 4HB components at crystal surfaces of single crystal aminolyzed, it was found that a small amount (up to ca. 2 mol%) of 4HB component can be incorporated into the P(3HB) mother crystal lattice irrespective of the 4HB content.     

Some Rheological Properties of Latex from Parthenium argentatum Gray Compared with Latex from Hevea brasiliensis and Ficus elastica

Latex was purified from Parthenium argentatum Gray (guayule), Hevea brasiliensis Müll. Arg. (the Brazilian or para rubber tree), and Ficus elastica Roxb. (the Indian rubber tree) in ammonium alginate at pH 10. The rheological properties of the different latices (rubber particle suspensions) were determined and compared using flow temperature ramps. Latex from all three species became more viscous with increasing rubber particle concentration and decreasing temperature. At any particular temperature and concentration, latex from F. elastica was by far the most viscous, whereas the H. brasiliensis latex was the least viscous. In addition, the tendency for the latex to coagulate increased with increasing temperature and increasing particle concentration. F. elastica latex was highly sensitive to temperature, H. brasiliensis latex was the least sensitive, and P. argentatum latex demonstrated intermediate properties. The underlying causes of these differences in latex rheology are not clear but may partially relate to the particle size (largest in F. elastica and smallest in H. brasiliensis), the particle size distribution, and/or to the considerable differences in the biochemical components of the monolayer biomembrane that surrounds the various rubber particles. Differences in the molecular weight of the rubber contained within the rubber particles seem less likely to play a role because the particles remain intact in this study.     

Degradation of a Terephthalate-containing Polyester by Thermophilic Actinomycetes and <Emphasis Type="Italic">Bacillus</Emphasis> Species Derived from Composts

We intended to find thermophilic degraders of terephthalate-containing Biomax^® films. Films in mesh bags were buried in composts (inside temperature: approximately 55–60 °C), resulting in the degradation of them in 2 weeks. Fluorescent microscopy of films recovered from composts showed that microorganisms gradually covered the surface of a film during composting. DGGE analysis of microorganisms on the composted film indicated the presence of Bacillus species as main species (approximately 80% of microbial flora) and actinomycetes (approximately 10–20%) as the second major flora. Isolation of Biomax^®-utilizing bacteria was focused on these two genera: two actinomycetes and one Bacillus species were isolated as pure best degraders from the composted polymer films, which were fragmented into small pieces. All the strains were thermophilic and identified, based on their 16S rDNA analyses. Degradation of polymer films was confirmed by (1) accelerated fragmentation of films in composts, compared with a control (no inoculum) and resultant decrease in molecular weights, (2) growth in a powdered Biomax^® medium, compared with a control without powdered Biomax^®, and (3) production of terephthalate in a powdered Biomax^® medium. In this way, we concluded that these bacteria were useful for degradation of thermostable Biomax^® products.