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DEFENSE POWERPOINT

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					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 packagingresponsible for the
    leaking/leeching out of styrene is.