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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 (SEM)




                      Janine Erica Padilla Dayao
                    Mary Bernadette Vallesfin Egloso




                             Submitted to the
                         Department of Biology
                      College of Arts and Sciences
                   University of the Philippines Manila
                    Padre Faura St., Ermita, Manila




                 In partial fulfillment of the requirements
                              for the degree of
                      Bachelor of Science in Biology
                                 March 2009
                                                           A Preliminary Study on the Potential……..ii
                                                                              Dayao and Egloso, 2009

                                    Department of Biology
                                 College of Arts and Sciences
                              University of the Philippines Manila
                               Padre Faura St., Ermita, Manila


                                     Announcement of
                              Undergraduate Thesis Presentation


                            JANINE ERICA PADILLA DAYAO
                         MARY BERNADETTE VALLESFIN EGLOSO


                                            Entitled


  A PRELIMINARY STUDY ON THE POTENTIAL BIODEGRADING ABILITY OF
  Xylaria sp. ON NATURAL RUBBER, CHICKEN FEATHER, AND POLYSTYRENE
              THROUGH SCANNING ELECTRON MICROSCOPY (SEM)


                                      For the degree of
                                Bachelor of Science in Biology


                               10:00 – 11:00AM, 13 March 2009
                                 Room 11-C, Rizal Hall (RH)
                                       CAS, U.P. Manila


          THESIS ADVISER                                        THESIS ADVISER
    Anna Theresa A. Santiago, M.P.H.                          Mary Ann Tavanlar, Ph.D.
          Assistant Professor
         Department of Biology                                 Antibiotic Laboratory
              U.P. Manila                                    BIOTECH, U.P. Los Baños


            THESIS READER                                         THESIS READER
         Kimberly S. Beltran, M.S.                                Jay T. Dalet, M.S.
               Instructor                                        Assistant Professor


Endorsed by:                                               Authorized by:


Samuel M. Go, M.S.P.H.                                     Marilen M. Parungao, M.Sc.
Chair                                                      Chair
Thesis Committee                                           Department of Biology
CAS, U.P. Manila                                           CAS, U.P. Manila
                                                         A Preliminary Study on the Potential……..iii
                                                                             Dayao and Egloso, 2009

                                   Department of Biology
                                College of Arts and Sciences
                             University of the Philippines Manila
                              Padre Faura St., Ermita, Manila




                                     ENDORSEMENT




       The thesis attached hereto, entitled “A Preliminary Study on the Potential Biodegrading
Ability of Xylaria sp. on Natural Rubber, Chicken Feather, and Polystyrene Through Scanning
Electron Microscopy (SEM)” prepared and submitted by Janine Erica Padilla Dayao and Mary
Bernadette Vallesfin Egloso, in partial fulfillment of the requirements for the degree of
Bachelor of Science in Biology was successfully defended on March 2009.




ANNA THERESA A. SANTIAGO, M.P.H.                           MARY ANN TAVANLAR, Ph.D.
         Thesis Adviser                                          Thesis Adviser




  KIMBERLY S. BELTRAN, M.S.                                      JAY T. DALET, M.S.
       Thesis Reader                                                Thesis Reader




       This undergraduate thesis is hereby officially accepted as partial fulfillment of the
requirements for the degree of Bachelor of Science in Biology.




MARILEN M. PARUNGAO, M.Sc.                              REYNALDO H. IMPERIAL, Ph.D.
Chair                                                   Dean
Department of Biology                                   College of Arts and Sciences
                                                      A Preliminary Study on the Potential……..iv
                                                                         Dayao and Egloso, 2009

                               BIOGRAPHICAL DATA


Name:               Mary Bernadette V. Egloso

Nickname:           Addie, Madette

Birthday:           June 20, 1988

Address:            135 Phase 4 Block 5 Lot 18, Our Lady of Lourdes Avenue., V&G
                    Subdivision, Tacloban City, Leyte

Telephone Number: (053) 327 – 3714

Mobile Number:      09153831788

E-mail Address:     adelaide17madette@yahoo.com

Parents:            Efren P. Egloso
                    Salve V. Egloso

Sibling:            Mary Jeane V. Egloso


Educational Background:
       Level                   School               Years Attended        Recognition
    Pre-School      United Church Family Life        1993 – 1996         Honor Student
                  Program Pre-Elementary School
     Primary          Sto. Niño SPED Center          1996 – 2001           Salutatorian
      School
    Secondary     Philippine Science High School     2001 – 2005
      School         – Eastern Visayas Campus
     College        University of the Philippines    2005 – 2009         College Scholar
                               Manila


Organizations/Affiliations:
      Member, UP Manila Chorale
      Member, Youth for Christ, UP Manila Chapter
      Member, UP Biological Sciences Society
      Member, UP Manila Quod Erat Demonstrandum
      Member, Biology Majors Association
                                                    A Preliminary Study on the Potential……..v
                                                                       Dayao and Egloso, 2009

                                BIOGRAPHICAL DATA


Name:                Janine Erica P. Dayao

Nickname:            Jana

Birthday:            October 4, 1988

Address:

Telephone Number:

Mobile Number:

E-mail Address:      jana_erica99@yahoo.com

Parents:

Sibling:


Educational Background:
             Level                School      Years Attended        Recognition
          Pre-School                           1993 – 1996
        Primary School                         1996 – 2001
       Secondary School                        2001 – 2005
            College                            2005 – 2009


Organizations/Affiliations:

        Member, UP Biological Sciences Society
        Member, UP Manila Quod Erat Demonstrandum
        Member, Biology Majors Association
                                                          A Preliminary Study on the Potential……..vi
                                                                             Dayao and Egloso, 2009

                                  ACKNOWLEDGMENTS


GOD:
        First and foremost, we would like to thank our Lord Jesus Christ for his guidance
throughout the entire process of our study, for sustaining and keeping us united, as He paved
every step of the way to the accomplishment of finishing our thesis.

Our Parents:
        To Daddy Mia and Mommy Bambi (Janine’s parents), and Papa Boy and Mama Salve
(Addie’s parents), we are so blessed for having you as our parents and guardians. Thank you
very much for the continuous moral and financial support, love and guidance, as we are far apart
from you. You are the main reason why we do our best in everything we do, in our academics,
and in our thesis in particular. We love you very much.

Asst. Prof. Anna Theresa Santiago:
       We would like to extend our heartfelt gratitude to our dear thesis adviser, Ma’am
Anna/Ata for her unwavering supervision, care, patience and moral support. We are so blessed to
have an ever understanding and supportive adviser as Ma’am Anna. Thank you very much
Ma’am.

Dr. Mary Ann Tavanlar:
        Thank you so much to Dr. Tavanlar, our thesis adviser in UPLB BIOTECH, for accepting
us as your thesis advisees and for your incessant guidance and patience, without whom we would
never have even achieved even just the first step in this thesis. Thank you also for teaching us
and helping us come up with a good and comprehensive manuscript, and for BIOTECH which
provided the materials needed for our study.

Prof. Jay Dalet:
       Sir Dalet, thank you so much for the patience, knowledge and time you have imparted in
meticulously reviewing our manuscript, without whom we would not have foreseen the errors
and grey areas in our study.

Prof. Kimberly Beltran:



Asst. Prof. Mark Romano:



UST and de La Salle Libraries:
                                                           A Preliminary Study on the Potential……..vii
                                                                               Dayao and Egloso, 2009




Blockmates and Friends:




                                          ABSTRACT


        The potential biodegrading ability of the Xylaria sp. strains on three pollutants, namely,
natural rubber, chicken feather and polystyrene were determined in terms of colonization. The
fungal strains namely the wild type, SDM (sterile dark mycelia) and the five mutant strains (PNL
114, PNL 116, PNL 118, E26 and E35) were cultured in a pH 5.0 mineral medium with 0.5%
glucose as carbon source, while the 3 pollutant substrates as co-carbon source, and stored at
250C. The initial weights of the pollutants were measured before inoculation. After 50 days of
incubation, the pollutants were removed from the medium and weights were again measured to
determine percent weight loss, a method that would measure utilization of the pollutant samples.
But instead of constant weight loss, weight gains resulted, which could be attributed to the
adherence and penetration of mycelia of the Xylaria sp. strains in the three pollutant substrates.
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. The black
variant strain E26 has shown the most potential for biodegrading natural rubber, followed by the
albino mutants. Among the black variant strains, E26 has shown the greatest biodegrading
potential for natural rubber, nonetheless the albino mutants also showed 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.
                                A Preliminary Study on the Potential……..viii
                                                     Dayao and Egloso, 2009




                  TABLE OF CONTENTS

TITLE PAGE…………….………………………………………………………………… i

BIOGRAPHICAL DATA………………………………………………………………… ii

ACKNOWLEDGMENTS………………………………………………………………… iv

TABLE OF CONTENTS………………………………………………….……………….                          vi

LIST OF FIGURES…………………………………………………………………………..                              vii

LIST OF TABLES…………………………………………………………………………                            viii

LIST OF APPENDICES……………………………………………………………………..                              ix

ABSTRACT………………………………………………………………………………….                                    x

INTRODUCTION…………………………………………………………………………...                                1

REVIEW OF RELATED LITERATURE………………………………………………………

MATERIALS AND METHODS……………………………………………………………….

RESULTS……………………………………………………………………………………….

DISCUSSION…………………………………………………………………………………...

CONCLUSION……………………………………………………………………….…………

RECOMMNEDATIONS……………………………………………………………………….

LITERATURE CITED………………………………………………………………………….
                                                         A Preliminary Study on the Potential……..ix
                                                                            Dayao and Egloso, 2009

APPENDIX……………………………………………………………………………………...




                                    LIST OF FIGURES



FIGURE                                                                                    PAGE

 1. Scheme for Polymer Biodegradation………………………………………………………..

 2. Natural rubber is a polymer called polyisoprene…………………………….………....……

 3. Gallus sp. is the source of feathers for the current study……………....……………………

 4. Polystyrene…………………………………………………………………………………..

 5. Growth of Xylaria sp. in mineral medium with 0.5% glucose and natural rubber…………..

 6. SEM of the non-inoculated natural rubber control without the fungus……………………...

 7. SEM of natural rubber surface colonized by the SDM or wild type strain
     after 50 days of incubation………………………………………………………………...

 8. SEM of natural rubber surface colonized by the PNL 114 mutant strain
     after 50 days of incubation………………………………………………………………...

 9. SEM of natural rubber surface colonized by the PNL 116 mutant strain
     after 50 days of incubation………………………………………………………………..

 10. SEM of natural rubber surface colonized by the PNL 118 mutant strain
      after 50 days of incubation……………………………………………………………….

 11. SEM of natural rubber surface colonized by the E26 mutant strain
      after 50 days of incubation……………………………………………………………….

 12. SEM of natural rubber surface colonized by the E35 mutant strain
                                                     A Preliminary Study on the Potential……..x
                                                                        Dayao and Egloso, 2009

     after 50 days of incubation……………………………………………………………….

13. Growth of Xylaria sp. in mineral medium with 0.5% glucose and chicken feather……….

14. SEM of non-inoculated chicken feather control without the fungus………………………

15. SEM of chicken feather colonized by the SDM wild type strain
     after 50 days of incubation……………………………………………………………….

16. SEM of chicken feather colonized by the PNL 114 mutant strain
     after 50 days of incubation……………………………………………………………….

17. SEM of chicken feather colonized by the PNL 116 mutant strain
     after 50 days of incubation……………………………………………………………….

18. SEM of chicken feather colonized by the PNL 118 mutant strain
     after 50 days of incubation……………………………………………………………...

19. SEM of chicken feather colonized by the E26 mutant strain
     after 50 days of incubation……………………………………………………………...

20. SEM of chicken feather colonized by the E35 mutant strain
     after 50 days of incubation……………………………………………………………...

21. Growth of Xylaria sp. in mineral medium with 0.5% glucose and polystyrene………….

22. SEM of non-inoculated control sample…………………………………………………..

23. SEM of polystyrene surface colonized by the SDM wild type strain
     after 50 days of incubation……………………………………………………………..

24. SEM of polystyrene surface colonized by the PNL 114 mutant strain
     after 50 days of incubation……………………………………………………………..

25. SEM of polystyrene surface colonized by the PNL 116 mutant strain
     after 50 days of incubation……………………………………………………………..

26. SEM of polystyrene surface colonized by the PNL 118 mutant strain
     after 50 days of incubation……………………………………………………………..

27. SEM of polystyrene surface colonized by the E26 mutant strain
     after 50 days of incubation……………………………………………………………..

28. SEM of polystyrene surface colonized by the E35 mutant strain
     after 50 days of incubation……………………………………………………………..
                                                     A Preliminary Study on the Potential……..xi
                                                                        Dayao and Egloso, 2009




                                  LIST OF TABLES



TABLE                                                                                 PAGE

 1. Comparative growth of the PNL mutants and wildtype SDM on MMG
       and mineral medium with various supplements. …………………………………….…

 2. Growth of the four albino mutants on mineral medium with and without
     supplements as compared to the wildtype SDM………………………………...…..……

 3. Growth of the two black mutants on mineral medium with supplements
       MDPE and PEG……………………………………………………………..….………

 4. Table of the Macroscopic/Physical and Microscopic Observations of the Xylaria sp.
       Mutant Strains and Wild Type on Colonized Natural Rubber after 50 days of
       Incubation…………………………………………………………………….………..

 5. Table of the Macroscopic/Physical and Microscopic Observations of the Xylaria sp.
       Mutant Strains and Wild Type on Colonized Chicken Feather after 50 days of
       Incubation…………………………………………………………………….………..

 6. Table of the Macroscopic/Physical and Microscopic Observations of the Xylaria sp.
       Mutant Strains and Wild Type on Colonized Polystyrene after 50 days of
       Incubation…………………………………………………………………….………..
……………….………..

				
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