Sterile distilled 70% EtOH Surface H20 (2x; 1min) (3mins) sterilization INOCULUM Removal of Incubation for fungi; gentle & 50 days even scraping INOCULUM Surface sterilization Sterile 70% EtOH distilled H20 (2x; 1min) (3mins) Xylaria sp. variant strains: SDM/Wild type, PNL 114, PNL 116, PNL 118, E26, E35 Mineral Plate/slant of Medium PDA stock culture (0.5% gluc; pH5;25˚C) 15mL Mineral Medium 0.5% Glucose 0.1M NaOH/ 0.1M HCl Substrate pollutant Natural rubber Substrate Chicken feather pollutant Polystyrene 2 ml Xylaria sp. SDM/Wild type 2 ml PNL 114 Xylaria sp. PNL 116 PNL 118 E35 E26 PNL114 PNL114 PNL114 SDM/ SDM/ Wild Wild type type PNL116 PNL116 E26 E26 PNL114 PNL114 PNL118 PNL118 E35 E35 SDM/ SDM/ NATURAL Wild Wild type type PNL116 PNL116 E26 E26 RUBBER PNL114 PNL114 PNL118 PNL118 E35 E35 SDM/ SDM/ CHICKEN Wild Wild type type PNL116 PNL116 E26 E26 FEATHER PNL114 PNL114 PNL118 PNL118 E35 E35 SDM/ SDM/ Wild Wild PNL116 PNL116 E26 E26 POLYSTYRENE type type PNL114 PNL114 PNL118 PNL118 E35 E35 NATURAL RUBBER CHICKEN FEATHER POLYSTYRENE RUN 1 RUN 2 RUN 3 mineral rinsed distilled rinsed 70% medium + water (2x;1min EtOH (1x;3min) fungi decanted each) gentle scraping Substrates cut to 0.5cm x 0.5cm pieces discovered by Cuevas and Manaligod (1997) growing on a sando plastic bag, buried in forest soil and litter in the lowland secondary forest of Mt. Makiling, Laguna comprised of sterile melanin pigmented mycelia reported as ascomycete sterile dark mycelia (ASDM). Cultural studies: Class Ascomycetes, Order Xylariales, Genus Xylaria. Clutario and Cuevas (2001) ◦ Xylaria sp. can utilize polyethylene plastic strips as alternative C source ◦ observed visible damages of the surface structure (tearing and striations caused by active burrowing) one of the most commonly encountered groups of ascomycetes stromatic, peritheciate, with an iodine-positive ascus apical ring, and with one-celled, dark ascospores on which a germination slit can be found. saprobic on decaying hardwood stumps and logs growing alone or, more commonly in clusters appearing in spring and not decaying until late summer or fall quiescent colonizers that decompose lignin and cellulose when a plant dies Some are associated with insect nests. many are plant pathogens. Many are endophytes. Commonly found throughout the temperate and tropical regions of the world. Xylaria sp. variant strains: SDM/Wild type, PNL 114, PNL 116, PNL 118, E26, E35 wild type SDM mutagenesis + protoplast fusion (thru NTG) determined morphological and biochemical characteristics or markers in the wild type and mutants High deposition of melanin in wild type SDM Results of Characterization Study (Tavanlar & Lat, 2008) less dependent on glucose in the medium for growth and hyphal tip extension. loss of melanin pigmentation improved ability to grow on reduced glucose levels tolerate 0.1% w/v copper hydroxide and 0.005% benomyl utilize 1% w/v polyethylene glycol 6000, Tween 80 and acetamide as source of carbon as compared to the wild type may potentially exhibit enhanced degradation of polyethylene plastics than the wild type may better survive environments with less available amounts of readily utilizable carbon sources such as the surface of plastics than the wild type from the latex of the Hevea brasiliensis also known as the rubber tree. mainly composed of cis-1,4 polyisoprene Average Composition of Latex Gloves from the Rubber Tree (in % (wt/wt)) 25-35% polyisoprene 0.5-0.6% polar lipids 1-1.8% protein 0.4-0.6% inorganic components 1-2% carbohydrates 0.4% amino acids, amides, etc. 0.4-1.1% neutral lipids 50-70% water rubber particles ~3- to 5- um covered by a layer of proteins and lipids ◦ serves to divide the hydrophobic rubber molecules from the hydrophilic environment. ◦ due to some allergic potential caused by Hevea proteins, methods to remove these proteins were applied classic subject of biodegradation studies ◦ high rate of its yearly manufacture which is several million tons, ◦ slow rate of natural degradation In the Philippines, chicken feathers aren’t a publicly recognized problem build-up in the environment and landfills result to future pollution problems and protein wastage accumulation could serve as a breeding ground for various harmful pathogens in the US, 2B lbs produced by the poultry industry most abundant protein not easily degraded tightly packed structural arrangement α-keratin/β-keratin Stability higher degree of cross-linking by disulfide bonds, hydrophobic interactions, and hydrogen bonds ◦ water-insoluble ◦ extremely resistant to biodegradation ◦ poorly susceptible to digestion by the most common peptidases like papain, trypsin and pepsin A group of proteolytic enzymes capable to hydrolyze insoluble keratins more efficiently than other proteases degrade the beta-keratin component microbes use feather as C, N, S and energy for their nourishment inducible whenever substrates of keratin composition are present an aromatic polymer and an inexpensive, hard plastic synthesized from the aromatic monomer styrene, from petroleum products. thermoplastic solid in room temperature or liquid when melted ◦ polymer chains are only weakly bonded (van der Waals forces) most common form EPS/Expanded Polystyrene/Styrofoam one of the packaging industry's toughest environmentally challenging products ◦ enormous sustaining longevity ◦ High durability but it is not biodegradable ◦ very difficult to recycle light weight property known neurotoxin and animal carcinogen mimics estrogen disrupt normal hormone functions (thyroid problems, menstrual irregularities, breast and prostate cancer) low platelet counts or hemoglobin values, chromosomal and lymphatic abnormalities EPS food packagingresponsible for the leaking/leeching out of styrene is.