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					                 A Preliminary Study on the Potential
                  Biodegrading Ability of Xylaria sp.
                 on Natural Rubber, Chicken Feather
                  and Polystyrene through Scanning
                         Electron Microscopy
                    Dayao, Janine Erica P.; Egloso, Mary Bernadtte V.;
                  Prof. Anna Theresa Santiago and Dr. Mary Ann Tavanlar




Dayao & Egloso
   (2009)
                       A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                              Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Introduction


Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                              Biodegradation
                 • breakdown of natural substances
                   through the action of enzymes
                   secreted by organisms
                 • natural and synthetic polymers
                   containing specific functional
                   groups, are potential substrates
                   for heterotrophic organisms
                 • organisms involved utilize or
                   metabolize these wastes as
                   sources of nutrients
Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Xylaria sp. wild type and
                          mutants



Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • 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
Dayao & Egloso
   (2009)          mycelia (ASDM).
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • 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
Dayao & Egloso     striations caused by active burrowing)
   (2009)
                   A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                          Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • 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
                 • quiescent colonizers that decompose
                   lignin and cellulose when a plant dies
                 • Some are associated with insect nests.
                 • many are plant pathogens and
                   endophytes.
                 • temperate and tropical regions of the
Dayao & Egloso
   (2009)
                   world.
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • 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
Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 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

Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Specific Objectives



Dayao & Egloso
   (2009)
                   A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                          Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • to determine if Xylaria sp. mutants
                   and wild type has the ability to
                   degrade through Scanning Electron
                   Microscopy

                 • to compare the biodegrading ability
                   of the wild type to each mutant

                 • to observe the macroscopic/physical
                   changes of the pollutants after
                   incubation with the Xylaria sp.
Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Significance of the
                        Study



Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • serve as a source of preliminary
                   information on the potential of Xylaria
                   sp. strains

                 • findings might be utilized in the
                   development of Xylaria sp. as a good
                   biodegrading agent

                 • may provide a possible means to
                   discover and understand other
                   important characteristics of Xylaria sp.
                   and its mutants
Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Methodology


Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                    Sterile
                                                     70% EtOH                                  Surface
                 distilled H20
                  (2x; 1min)                          (3mins)                                sterilization




                    SEM
Dayao & Egloso
   (2009)
                      A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                             Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                                 washed



                 SEM                               SEM


Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 70% EtOH                             Surface
                  (3mins)                           sterilization



                    Sterile
                 distilled H20                              SEM
                  (2x; 1min)
Dayao & Egloso
   (2009)
                      A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                             Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Xylaria sp. variant strains:
                   SDM/Wild type, PNL 114, PNL
                   116, PNL 118, E26, E35




Dayao & Egloso
   (2009)
                   A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                          Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                                        Mineral
    Plate/slant
                                                        Medium
     of stock              PDA
                                                      (0.5% gluc;
      culture
                                                       pH5;25˚C)




Dayao & Egloso
   (2009)
                  A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                         Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                              15mL
                             Mineral
                             Medium




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                                       0.5%
                                                       Glucose




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                              0.1M
                             NaOH/
                            0.1M HCl




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                                    Substrate
                                                    pollutant




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                                                                Natural rubber
                                                    Substrate
                                                                                 Chicken feather
                                                    pollutant
                                                                                 Polystyrene




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                         2 ml
                      Xylaria sp.




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                                                               SDM/Wild type

                                                       2 ml                    PNL 114
                                                    Xylaria sp.
                                                                               PNL 116

                                                                               PNL 118

                                                E35                 E26




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                                    PNL114




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                              PNL114               PNL114




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                  SDM/         SDM/
                  Wild         Wild
                  type         type               PNL116         PNL116                  E26            E26




                 PNL114      PNL114               PNL118         PNL118                  E35            E35




Dayao & Egloso
   (2009)
                   A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                          Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • NATURAL
                                                        SDM/        SDM/
                                                        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




Dayao & Egloso                                         PNL114     PNL114       PNL118      PNL118        E35       E35
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
          NATURAL RUBBER CHICKEN FEATHER                                              POLYSTYRENE


RUN 1




RUN 2




RUN 3




 Dayao & Egloso
    (2009)
                   A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                          Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Substrates
                   cut to 0.5
                   x0.5cm
                   pieces




Dayao & Egloso
   (2009)
                         A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                                Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Natural Rubber


Dayao & Egloso
   (2009)
                   A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                          Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                    Macroscopic
                  before                                                    after




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 SDM wild type                                E26                                       E35




Dayao & Egloso    PNL 114                                  PNL 116                                PNL 118
   (2009)
                      A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                             Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
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Dayao & Egloso            my
   (2009)
                      A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                             Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 hy
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                            my




Dayao & Egloso   hy
   (2009)
                       A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                              Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Natural Rubber


                 • Hevea brasiliensis a.k.a rubber
                   tree
                 • cis-1,4 polyisoprene
                 • Biodegradation of rubber slow
                   process
                        Rose & Steinbuchel (2005) and
                   Bereeka (2006)
Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • SEM pictures:
                    fungi have adhered closely to the
                   substrate and grew on it.
                    substrate is now a combination of the
                   original pollutant which is natural rubber
                   and the Xylaria strains.

                 • Even after gentle scraping and washing
                   with 70% ethanol and water,
                        the mycelia have remained
                   embedded on the surface of the
                   substrate.
                       proof of non-aggregation on the
                   surface of the natural rubber.
Dayao & Egloso
   (2009)
                      A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                             Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 •Percent weight loss was not used as a method
                 due to weight gains
                  Strain   Initial   Final       Difference   Initial   Final     Difference    Initial   Final       Difference
                           Weight    Weight                   Weight    Weight                  Weight    Weight


                                     First Run                          Second Run                        Third Run

                  SDM      0.1909    0.1883       0.0026      0.1976    0.1929       0.0047     0.231     0.3216         -0.0906
                  SDM      0.1427    0.1489      -0.0062      0.2581    0.2535       0.0046     0.3524    0.3555         -0.0031
                   114     0.2041    0.2221       -0.018      0.3179    0.3117       0.0062     0.3942    0.3595         0.0347
                   114     0.1183    0.1408      -0.0225      0.2741    0.2693       0.0048     0.2766    0.2472         0.0294
                   116     0.1402    0.1363       0.0039      0.2797    0.2842       -0.0045    0.276     0.3283         -0.0523
                   116     0.2067    0.254       -0.0473      0.3771    0.3721       0.005      0.2748    0.2864         -0.0116
                   118     0.2002    0.197        0.0032      0.2853    0.2806       0.0047     0.2867    0.2202         0.0665
                   118     0.149     0.2088      -0.0598      0.2915    0.2814       0.0101     0.2879    0.2709         0.017
                  E26      0.1891    0.1851       0.004       0.2732    0.2698       0.0034     0.3284    0.2687         0.0597
                  E26
                           0.2106    0.204        0.0066      0.309     0.3045       0.0045     0.2458    0.2738         -0.028
                  E35      0.1183    0.1409      -0.0226      0.3229    0.3191       0.0038     0.3541    0.3877         -0.0336
                  E35      0.2009    0.2098      -0.0089      0.2232    0.2162       0.007      0.3514    0.3497         0.0017
Dayao & Egloso
   (2009)
                           A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                                  Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Percent weight loss was not used as a method
                 due to weight gains
                 Strain   Initial   Final       Difference   Initial   Final     Difference    Initial   Final       Difference
                          Weight    Weight                   Weight    Weight                  Weight    Weight



                                    First Run                          Second Run                        Third Run

                 SDM      0.1909    0.1883       0.0026      0.1976    0.1929       0.0047     0.231     0.3216         -0.0906
                 SDM      0.1427    0.1489      -0.0062      0.2581    0.2535       0.0046     0.3524    0.3555         -0.0031
                  114     0.2041    0.2221       -0.018      0.3179    0.3117       0.0062     0.3942    0.3595         0.0347
                  114     0.1183    0.1408      -0.0225      0.2741    0.2693       0.0048     0.2766    0.2472         0.0294
                  116     0.1402    0.1363       0.0039      0.2797    0.2842       -0.0045    0.276     0.3283         -0.0523
                  116     0.2067    0.254       -0.0473      0.3771    0.3721       0.005      0.2748    0.2864         -0.0116
                  118     0.2002    0.197        0.0032      0.2853    0.2806       0.0047     0.2867    0.2202         0.0665
                  118     0.149     0.2088      -0.0598      0.2915    0.2814       0.0101     0.2879    0.2709         0.017
                 E26      0.1891    0.1851       0.004       0.2732    0.2698       0.0034     0.3284    0.2687         0.0597
                 E26      0.2106    0.204        0.0066      0.309     0.3045       0.0045     0.2458    0.2738         -0.028
                 E35      0.1183    0.1409      -0.0226      0.3229    0.3191       0.0038     0.3541    0.3877         -0.0336
                 E35      0.2009    0.2098      -0.0089      0.2232    0.2162       0.007      0.3514    0.3497         0.0017
Dayao & Egloso
   (2009)
                          A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                                 Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • 0.5 % glucose + on the mineral medium
                       only initiated growth and
                   sustained the fungal growth for three
                   weeks. (Tavanlar, personal communication, march 9,2009)


                 • study of Tavanlar and Lat (2008)
                        wherein growth of Xylaria was
                   seen and measured upon its utilization
                   of mineral oil and PEG as alternative
                   carbon sources.

Dayao & Egloso
   (2009)
                       A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                              Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • The adherence and growth of the
                   fungus signified colonization on the
                   substrate.
                        formation of mycelial mats and
                   through the presence of mycelia,
                   spores and hyphae.

                 • According to the studies of Linos et al.
                   (2000), colonization is the initial step in
                   the biodegradation of polymers.


Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Fungal mechanism of natural
                   rubber biodegradation have not
                   yet been elucidated.
                 • But microbial rubber-degraders:
                 oxidative cleavage of the double
                   bonds in the polyisoprene chain
                   :Gordonia sp.
                 oligomers of aldehyade and
                   ketone groups were recovered.

Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Observed biofilm formation in the form of
                   mucilaginous sheaths
                 • Xylaria strains forms biofilm
                         (M.A Tavanlar,personal communication, March 9,2008)


                 • BIOFILM
                 mechanism by which microorganisms
                   secrete proteins and carbohydrates for
                   their survival in environments which are
                   low in nutrients
                 indicate that there is solid substrate
                   utilization by the microorganisms.
                 regarded as an initial step in
                   biodegradation.
                          (Morton & Surman, 1994; Linos et al., 2000).
Dayao & Egloso
   (2009)
                       A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                              Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Colonization
                 happens when the cells of the
                  organism secretes extracellular
                  polysaccharide substances (EPS).
                 results in the destruction of quality
                  or performance of the material.
                  fungi and bacteria have been
                  shown to attack and form biofilms
                  on natural rubber substrates.
                   (Morton & Surman, 1994; Linos et al., 2000).
Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • In general, the black Xylaria sp.
                   mutant strains, especially the E26
                   strain, have showed the greatest
                   biodegrading potential when
                   compared to the albino strains
                   namely PNL 114, 116 and 118.
                   But among the albino  114

                 • suggests that presence of melanin
                   in the black strains’ hyphal
                   structures could be playing a role
                   in biodegradation
Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • According to the study of Tavanlar and
                   Lat (2008)
                  white mutants possess thinner hyphal
                   extension tip due to the loss of melanin
                   structure around it; whereas the
                 black mutants have thicker hyphae due
                   to melanin around there structure.

                 • melanin
                 protection and resistance against the
                   pressure/friction of the substrate during
                   hyphal tip adherence and penetration
Dayao & Egloso        (M. A.Tavanlar, personal communication, March 4, 2009)
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • slow colonization
                   suggest that colonization
                   impedance due to
                   chemicals in the old latex
                   gloves used might be
                   taking place
                  (Bereeka et al.,2000 and Rose & Steinbuchel, 2005)




Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Chicken Feathers



Dayao & Egloso
   (2009)
                   A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                          Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                    Macroscopic
                  before                                                    after




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 SDM wild type                             E26                                       E35




Dayao & Egloso      PNL 114                                PNL 116                               PNL 118
   (2009)
                      A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                             Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
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Dayao & Egloso                                                                                                      br
   (2009)
                                                                         my
                      A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                          bl
                             Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
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Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Chicken Feathers
                 • 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
Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 CF = 90% keratin
                 • 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
Dayao & Egloso
                     pepsin
   (2009)
                      A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                             Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                  Keratinase
                 • 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
Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                        Polystyrene




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                    Macroscopic
                  before                                                    after




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 SDM wild type                        E26                                      E35




                    114                                   116                                     118
Dayao & Egloso
   (2009)
                      A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                             Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                                                         sh


                                                                               my



                                                                    sp                      hy
                                                                                                               sp



                                          d




                   my
   my



                                         h
                                                                                                               my


                                                              my
Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                                                       my




                                                                         sp




Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Polystyrene
                 • 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
Dayao & Egloso
   (2009)
                   Polystyrene/Styrofoam
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Polystyrene


                 • 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
Dayao & Egloso
   (2009)
                   property
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • 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.
Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Conclusion


Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Scanning Electron Microscopy (SEM)
                   revealed colonization of Xylaria sp.
                   strain on natural rubber, chicken
                   feather and polystyrene.

                 • Generally, the black strains, namely the
                   SDM wild type, E26 and E35, have
                   been observed to possess a higher
                   potential to biodegrade natural rubber,
                   chicken feather and polystyrene than
                   the albino mutants PNL 114, 116 and
                   118.
Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Among the black variant strains, E26 has
                   shown the greatest biodegrading potential for
                   natural rubber, nonetheless the albino
                   mutants also showed a potential to
                   biodegrade this substrate.

                 • Polystyrene was effectively colonized by the
                   three black strains SDM wild type, E26 and
                   E35, nonetheless the albino mutants also
                   showed potential ability to biodegrade this
                   substrate.

                 • Chicken feathers were colonized by the SDM
                   and E35 but may require a longer incubation
                   period for degradation
Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 Recommendations



Dayao & Egloso
   (2009)
                 A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                        Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • longer incubation time
                 • further taxonomic identification of
                   Xylaria strains
                 • test methods and standards for
                   biodegradable polymers:
                 Modified sturm test
                 closed bottle test
                 Petri dish screen
                 environmental chamber method
Dayao & Egloso
                 soil burial test
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Structural modifications can be
                   monitored in depth through infra-red
                   (IR) spectroscopy.

                 • The actual utilization or “absorption”
                   of carbon by the mycelia can be
                   computed.



Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Changes in molecular weight can be
                   checked by using high-temperature
                   size exclusion chromatography (ht-
                   SEC).

                 • Other techniques to evaluate
                   macroscopic modifications aside from
                   electron microscopy can be used, such
                  photonic microscopy
                 polarization microscopy
                 atomic force microscopy
Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Internal biodegradation can be
                   evaluated by change of rheological
                   properties:

                 Tensile strength is measured with a
                  tensile tester
                 elongation at break by a mechanical
                  tester
                 elongation percentage and elasticity by
                  dynamic mechanical thermal analysis.
Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Natural rubber
                 tests that would further confirm
                   presence of intermediate and by-
                   product compounds (staining of schiff’s
                   reagent and FTR-AITR spectroscopy)
                 treatment of natural rubber gloves
                   before subjecting it to biodegradation

                 • Polystyrene
                  Testing other grades of polystyrene to
                   be biodegraded.
Dayao & Egloso
   (2009)
                     A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                            Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy
                 • Feathers
                 To verify the degradation of
                   feather, more advanced tests
                   should be conducted wherein the
                   presence of soluble proteins and
                   amino groups concentration will be
                   observed.
                 Study the possibility that Xylaria
                   sp. can produce enzymes such as
                   keratinase, proteinase. Testing of
                   white feathers is also suggested
Dayao & Egloso
   (2009)
                    A Preliminary Study on the Potential Biodegrading Ability of Xylaria sp. on Natural Rubber,
                           Chicken Feathers, and Polystyrene through Scanning Electrons Microscopy

				
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