Uptake and Metabolism of TNT and GTN by Plants Expressing Bacterial Pentaerythritol Tetranitrate Reductase

The manufacture and improper disposal of explosives has resulted in a significant amount of land requiring remediation. The compound 2,4,6-trinitrotoluene (TNT) is the most persistent and toxic of the explosive pollutants with current treatment methods being energy intensive and costly. Bacterial enzymes such as pentaerythritol tetranitrate reductase (PETNR) from Enterobacter cloacae PB2 have been found to have activity against TNT; however, microbes often lack the biomassnecessary for remediation applications. The PETNRgene (onr1) was transformed into tobacco plants in an attempt to combine the metabolic diversity of microbes with the sequestering properties of plants. The resulting transgenic plants were shown to have enhanced tolerance to TNT during germination and as seedlings. Phytoremediation applications with these plants may provide an alternative treatment of TNT contamination.     

Assessing the Phytoremediation Potential of Tall Fescue and Sericea Lespedeza for Organic Contaminants in Soil

The phytoremediation potential of using tall fescue (Festuca arundinacea Schreb.) grass and sericea lespedeza (Lespedeza cuneata [Dum. ‐Cours.]) legume species was assessed using three different groups of organic contaminants in soil. One hundred parts per million (ppm) each of a nitroaromatic compound (TNT), a polycyclic aromatic hydrocarbon (Pyrene), and a polychlorinated biphenyl (Aroclor 1248) were used to contaminate the soils. The experiments were conducted using soils with high and low organic‐matter content. The results indicate that recoveries of Pyrene and TNT were very low in all treatments in soil with high organic‐matter content (6.3 percent) compared with recoveries in soil with low organic‐matter content (2.6 percent). In contrast, recoveries of PCB from soil were not dependent on the soil's organic‐matter content. Planting both the legume and grass species had significant effect on the transformations of TNT and PCB in the soil with low organic‐matter content and did not affect the fate of Pyrene in both soils. The amount of TNT transformed in the four months of plant growth was 63 percent in the tall fescue and 46 percent in the sericea‐planted soils, compared with only a 15 percent unaccounted loss in the unplanted control soils. Furthermore, the grass species, with its massive root system, was significantly better at causing TNT dissipation compared with the legume species, which has less root vegetative mass. The plant biomass, particularly the shoot weight of the tall fescue grass, was significantly increased as a result of TNT treatment. Tall fescue and sericea biomass did not appear to have any significant effect on Pyrene transformation. Planting sericea provided a significantly high level of PCB transformation in soils with either high or low amounts of organic matter. Tall fescue did not appear to have any significant effect on PCB transformation. © 2002 Wiley Periodicals, Inc.     

A Study of the Use of Alfalfa (Medicago sativa L.) for the Phytoremediation of Organic Contaminants in Soil

This article presents the results of a study that was conducted to determine the effectiveness of using alfalfa (Medicago sativa L.) to enhance the phytoremediation of three different types of chemical contaminants. The chemicals studied were trinitrotoluene (TNT), the polycyclic aromatic hydrocarbon (PAH) pyrene, and the polychlorinated biphenyl (PCB) Aroclor 1248. Experiments were conducted using soils that contained high and low organic matter content. The results indicated that recoveries of pyrene and TNT from soil were highly dependent on the soil organic matter content, while the recovery of PCB was not. Significantly low levels of pyrene and TNT were recovered from all treatments in the soil with 6.3 percent organic matter content compared to recovery levels found in soil with 2.6 percent organic matter. The presence of alfalfa plants had a significant effect on the transformation of TNT and PCB in the low organic matter content soil only and had no effect on the fate of pyrene. In the low organic matter soil, only 15 percent and 17 percent of the initial TNT and PCB levels, respectively, were transformed in the unplanted control soils compared to 66 percent and 77 percent in the alfalfa planted pots. In both soil types, pyrene dissipation could not be attributed to the presence of alfalfa plants. Overall, it was concluded that under high soil organic matter conditions, adsorption and covalent binding to the soil organic matter appeared to be the dominant force of pyrene and TNT removal. The effectiveness of using alfalfa to enhance PCB and TNT transformations was more significant in the lower organic matter soil; thus phytoremediation had a greater effect in soils with lower organic matter content. © 2001 John Wiley & Sons, Inc.     

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.     

Biochemical Cooperation Between Klebsiella oxytoca Sc and Methylobacterium mesophilium Sr for Complete Degradation of Dimethyl Isophthalate

Klebsiella oxytoca Sc and Methylobacterium mesophilicum Sr were isolated from enrichment cultures using dimethyl isophthalate (DMI) as the sole carbon and energy source and mangrove sediment as an inoculum. Complete degradation of DMI required both species of bacteria at different biochemical transformation steps. The biochemical degradation pathway was DMI to monomethyl isophthalate (MMI) by K. oxytoca Sc, MMI to isophthalate (IPA) by M. mesophilium Sr, and IPA by both K. oxytoca Sc and M. mesophilium Sr sequentially. The consortium comprising of K. oxytoca Sc and M. mesophilicum Sr was effective in complete degradation of DMI in eight days. Our results suggest that a consortium of microorganisms indigenous to the mangrove environment is responsible for mineralization of environmental pollutant DMI through biochemical cooperation.     

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.     

Sorting efficiency and combustion properties of municipal solid waste during bio-drying

One aerobic and two combined bio-drying processes were set up to investigate the quantitative relationships of sorting efficiency and combustion properties with organics degradation and water removal during bio-drying. Results showed that the bio-drying could enhance the sorting efficiency of municipal solid waste (MSW) up to 71% from the initial of 34%. The sorting efficiency was correlated with water content negatively (correlation coefficient, r = −0.89) and organics degradation rate positively (r = 0.92). The higher heating values (HHVs) were correlated with organics degradation negatively for FP (i.e. the sum of only food and paper) (r = −0.93) but positively for the mixing waste (MW) (r = 0.90), whereas the lower heating values (LHVs) were negatively correlated with water content for both FP (r = −0.71) and MW (r = −0.96). Other combustion properties depended on organics degradation performance, except for ignition performance and combustion rate. The LHVs could be greatly enhanced by the combined process with insufficient aeration during the hydrolytic stage. Compared with FP, MW had higher LHVs and ratios of volatile matter to fixed carbon. Nevertheless, FP had higher final burnout values than MW.     

Synthesis of copolyesters containing poly(ethylene terephthalate) and poly(ε-caprolactone) units and their susceptibility toPseudomonas sp. lipase

Copolyesters containing poly(ethylene terephthalate) (PET) and poly(-caprolactone) (PCL) were synthesized from PET and PCL homopolymers by transesterification reaction at 270°C in the presence of catalyst. The copolyesters were characterized by¹³C-NMR and differential scanning calorimetry (DSC). The degradation behavior of PCL byPseudomonas sp. lipase in buffer solution (pH 7) and tetrahydrofuran (THF) was investigated by gel permeation chromatography (GPC) and¹H-NMR. From these experiments, it was found thatPseudomonas sp. lipase acted endoenzymatically on PCL. Using this lipase, degradation tests for PET/PCL copolyesters whose PCL content was below 50% by weight were also performed in buffer solution (pH 7). However, evenPseudomonas sp. lipase with high degradation activity on PCL did not easily degrade the PCL unit in PET/PCL copolyesters.     

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.     

Factors affecting polycyclic aromatic hydrocarbon biodegradation by Aspergillus flavus

This study investigated the ability of fungi isolated from highly contaminated soil to biodegrade polycyclic aromatic hydrocarbon (PAH) compounds, as well as the effect of several parameters on the biodegradation ability of these fungi. The isolated fungi were identified using ITS rDNA sequencing and tested using 2,6‐dichlorophinolendophenol to determine their preliminary ability to degrade crude oil. The top‐performing fungi, Aspergillus flavus and Aspergillus fumigatus, were selected to test their ability to biodegrade PAH compounds as single isolates. After 15 days of incubation, A. flavus degraded 82.7% of the total PAH compounds, with the complete degradation of six compounds, whereas A. fumigatus degraded 68.9% of the total PAHs, with four aromatic compounds completely degraded. We also tested whether different temperatures, pH, and nitrogen sources influenced the growth of A. flavus and the degradation rate. The degradation process was optimal at a temperature of 30°C, pH of 5.5, and with nitrogen in the form of yeast extract. Finally, the ability of the fungal candidate, A. flavus, to degrade PAH compounds under these optimum conditions was studied. The results showed that 95.87% of the total PAHs, including 11 aromatic compounds, were completely degraded after 15 days of incubation. This suggests that A. flavus is a potential microorganism for the degradation of PAH compounds in aqueous cultures.     

Effects of Temperature and Relative Humidity on Polylactic Acid Plastic Degradation

Three high molecular weight (120,000 to 200,000 g mol^–1) polylactic acid (PLA) plastic films from Chronopol (Ch-I) and Cargill Dow Polymers (GII and Ca-I) were analyzed for their degradation under various temperature and relative humidity (RH) conditions. Two sets of plastic films, each containing 11 samples, were randomly hung in a temperature/humidity-controlled chamber by means of plastic-coated paper clips. The tested conditions were 28, 40, and 55°C at 50 and 100% RH, respectively, and 55°C at 10% RH. The three tested PLA films started to lose their tensile properties when their weight-average molecular weight (M _w) was in the range of 50,000 to 75,000 g mol^–1. The average degradation rate of Ch-I, GII, and Ca-I was 28,931, 27,361, and 63,025 M _w/week, respectively. Hence, GII had a faster degradation rate than Ch-I and Ca-I under all tested conditions. The degradation rate of PLA plastics was enhanced by the increase in temperature and relative humidity. This trend was observed in all three PLA plastics (Ca-I, GII, and Ch-I). Of the three tested films, Ch-I was the first to lose its mechanical properties, whereas Ca-I demonstrated the slowest loss, with mechanical properties under all tested conditions.     

Aging of Toughened Polylactic Acid Nanocomposites: Water Absorption,Hygrothermal Degradation and Soil Burial Analysis

The environmental aging behaviour of montmorillonite (MMT) filled polylactic acid (PLA) nanocomposites (PLA/MMT) and linear low density polyethylene (LLDPE)-toughened PLA (PLA/LLDPE ratio = 90/10) nanocomposites (PLA/LLDPE/MMT) were investigated in this study. The nanocomposites were subjected to water absorption, hygrothermal degradation and soil burial analysis. Both PLA/MMT and PLA/LLDPE/MMT nanocomposites were immersed in distilled water at three different temperatures (room temperature, 60, and 90 °C) and the weight difference before and after immersion was calculated. The kinetics of water absorption for both nanocomposites followed the Fick’s second law of diffusion, where a linear relationship exists between the initial moisture absorption at any time t and t ^1/2 (the square root of time), followed by a horizontal plateau (saturation). The equilibrium moisture content (M _m ) and diffusion coefficient (D) of PLA nanocomposites increased with the addition of MMT (2 phr) and LLDPE. However, the D values of both nanocomposites decreased by increasing MMT (4 phr). The M _m for PLA/MMT and PLA/LLDPE/MMT nanocomposites increased by increasing immersion temperature (60 °C) and prolonged immersion resulted in hygrothermal degradation of both nanocomposites. The hygrothermal degradation studies showed that PLA degrades much faster at 90 °C as compared to 60 °C in both the nanocomposites. The addition of MMT and LLDPE improved the hygrothermal stability of PLA in both nanocomposites. Soil burial test revealed deterioration of impact strength in all samples while the rate of biodegradation was retarded in the presence of MMT and LLDPE.     

Effects of Electron-Beam Irradiation and Ultraviolet Light (365 nm) on Polylactic Acid Plastic Films

Strips of Ca-I [polylactic acid (PLA) monolayer plastic films from Cargill Dow Polymers LLC, Minnetonka, MN] cut in the machine and nonmachine directions were irradiated with an electron beam using a CIRCLE III Linear Accelerator (MeV Industries S.A., Jouy-en-Josas, Cedex, France). The effects of 33-kGy irradiation on the physical properties of the Ca-I strips were studied. In addition, the effects of ultraviolet (UV) light (365-nm) illumination on the degradation of three PLA plastic films, Ch-I (PLA monolayer plastic films from Chronopol, Golden, CO), GII (PLA trilayer plastic films from Cargill Dow Polymers LLC), MN), and Ca-I (PLA monolayer plastic films from Cargill Dow Polymers LLC) were analyzed by a modified ASTM D5208-91 method. Results showed that irradiation had decreased the weight-average molecular weight (M _w), stress at break, percentage of elongation, and strain energy of Ca-I by 35.5, 26.7, 32.3, and 44.8%, respectively (P < 0.01). The shelf life of the irradiated Ca-I strips at 5°C and <20 ± 5% RH was about 6 months. The degradation rate of Ch-I, GII, and Ca-I with no UV light treatment at 55°C and 10% RH was 2512, 5618, and 3785 M _w/week, respectively. Under the UV light illumination (365 nm), the degradation rate of Ch-I, GII, and Ca-I, was 2982, 8722, and 7467 M _w/week, respectively. Hence, the degradation rate of GII and Ca-I was increased 55 and 97% by UV light (P < 0.008), respectively. This trend was not observed in Ch-I because its starting M _w (78,000 g/mol) was close to the tensile strength lost range (50,000 to 75,000 g/mol) of PLA films. To our knowledge, this is the first study to demonstrate that UV light does further enhance the degradation of PLA films.     

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.     

Transport and Immobilization of 2,4,6 15N-Trinitrotoluene in Soil Microcosms Subjected to Long Term Incubation Under Aerobic Conditions

¹⁵N-labeling and solid-state ¹³C and ¹⁵N nuclear magnetic resonance (NMR) spectroscopy was applied to study the immobilization of 2,4,6 trinitrotoluene (TNT) into soil organic matter (SOM). Uncontaminated soil from the Ap horizon of a Luvisol was mixed with ¹⁵N-TNT (enrichment: 99 atm%) and laid over an unspiked layer of the same material. The latter covered soil from the Bt horizon. The microcosms were aerobically incubated under laboratory conditions for up to 11 months. After 1 week, within the total microcosm approximately 90% of the added ¹⁵N (¹⁵N_add) were recovered, mostly in the top layer (87%). After 11 months, this amount decreased to 71%, indicating losses due to denitration or transamination. Within two months, half of ¹⁵N_add had been immobilized in the residues not extractable with organic solvents and water. The amount of the sequestered ¹⁵N_add remained fairly constant until the end of the experiment pointing towards a high stability of TNT-SOM associates. Solid-state ¹⁵N NMR revealed their formation by covalent binding, most tentatively as amides. Complete reduction of TNT to triaminotoluene (TAT) was not prerequisite. The most pronounced downwards movement of ¹⁵N-TNT occurred during the first two months. The major part of it, however, experienced quick immobilization, leaving approximately 10% of ¹⁵N_add recovered in the leachate at the end of the experiment. Calculations indicated contributions of inorganic ¹⁵N_add. Approximately 25% of its organic ¹⁵N_add originated from condensed N, suggesting that in soils the transport of partly reduced TNT is in close association with the organic matter of the soil solution to which they are covalently bound.     

Biodegradability of Chemically Modified Starch (RS4)/PVA Blend Films: Part 2

Biodegradable films were successfully prepared by using cornstarch (CS), chemically modified starch (RS4), polyvinyl alcohol (PVA), glycerol (GL), and citric acid (CA). The physical properties and biodegradability of the films using CS, RS4, and additives were investigated. The results of the investigation revealed that the RS4-added film was better than the CS-added film in tensile strength (TS), elongation at break (%E), swelling behavior (SB) and solubility (S). Especially, the RS4/PVA blend film with CA as an additive showed physical properties superior to other films. Furthermore, when the film was dried at low temperature, the properties of the films clearly improved because the hydrogen bonding was activated at low temperature. The biodegradation of films was carried out using the enzymatic, microbiological and soil burial test. The enzyme used in this study was amyloglucosidase (AMG), α-amylase (α-AM) and β-amylase (β-AM). At the enzymatic degradation test, the GL-added films had an approximately 60% degradation, while the CA-added films were degraded about 25%. The low degradation value on CA-added film is attributed to low pH of film added CA that deactivated the enzymatic reaction. The microbiological degradation teat was performed by using Bacillus subtilis and Aspergillus niger.     

The Effect of CTBN Concentrations on the Kinetic Parameters of Decomposition of Blends of Epoxy Resins Modified with Carboxyl-Terminated Liquid Copolymer

Diglycidyl ether of bisphenol—A (DGEBA)—based epoxy resin was blended in the ratio of 3:1 (weight basis) with cycloaliphatic epoxy (CAE) resin. The prepared blend sample was further blended with different weight percentages of carboxyl-terminated butadiene acrylonitrile copolymer (CTBN) ranging between 0 and 25 wt% with an interval of 5 wt% and cured with stiochiometric amounts of 4, 4’- diamino diphenyl sulphone (DDS) cure agent. Structural changes during blending were studied by Fourier-transform infra-red (FTIR) spectroscopic analysis. The kinetic parameters, viz., order of decomposition reaction (n), activation energy (E), pre-exponential factor (Z) and rate decomposition constant (k), for the decomposition of the samples were calculated by applying Coats-Redfern equation over thermogravimetric (TG) data. The degradation of each sample followed second-order degradation kinetics, which was calculated by Coats-Redfern equation using best-fit analysis. This was further confirmed by linear regression analysis. The validity of data was checked by t-test statistical analysis. Further, the blend sample had higher initial degradation temperature and activation energy than its respective pure epoxy resin indicating that the CTBN acted as thermal stabilizer for epoxy resin which improved the thermal stability.     

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.     

Degradation of Polylactic Acid (PLA) Plastic in Costa Rican Soil and Iowa State University Compost Rows

In this study the degradation of polylactic acid (PLA) plastic films in Costa Rican soil and in a leaf composting environment was investigated. Three types of PLA films were used: Ch-I, (PLA monolayer plastic films from Chronopol, Golden, CO), GII (PLA trilayer plastic films from Cargill Dow Polymers LLC, Minnetonka, MN), and Ca-I (PLA monolayer plastic films from Cargill Dow Polymers LLC). The average soil temperature and moisture content in Costa Rica were 27°C and 80%, respectively. The average degradation rate of PLA plastic films in the soil of the banana field was 7675 M _w/week. Two compost rows were set up at the Iowa State University (ISU) (Ames) compost site. Temperature and relative humidity of the compost rows were kept at 55 ± 5°C and 50 ± 10% RH, respectively. The degradation rates of GII and Ca-I in the compost rows were 113,290 and 71,283 M _w/week, respectively. Therefore, it was estimated that in Costa Rican soil and in compost rows, PLA would be visibly degraded in 6 months and in 3 weeks, respectively.     

Fungal Degradation of the Bioplastic PHB (Poly-3-hydroxy- butyric acid)

PHB (poly-3-hydroxybutyric acid) is a thermoplastic polyester synthesized by Ralstonia eutropha and other bacteria as a form of intracellular carbon and energy storage and accumulated as inclusions in the cytoplasm of these bacteria. The degradation of PHB by fungi from samples collected from various environments was studied. PHB depolymerization was tested in vials containing a PHB-containing medium which were inoculated with isolates from the samples. The degradation activity was detected by the formation of a clear zone below and around the fungal colony. In total, 105 fungi were isolated from 15 natural habitats and 8 lichens, among which 41 strains showed PHB degradation. Most of these were deuteromycetes (fungi imperfecti) resembling species of Penicillium and Aspergillus and were isolated mostly from soils, compost, hay, and lichens. Soil-containing environments were the habitats from which the largest number of fungal PHB degraders were found. Other organisms involved in PHB degradation were observed. A total number of 31 bacterial strains out of 67 isolates showed clear zones on assay medium. Protozoa, possible PHB degraders, were also found in several samples such as pond, soil, hay, horse dung, and lichen. Lichen, a fungi and algae symbiosis, was an unexpected sample from which fungal and bacterial PHB degraders were isolated.