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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
Janine Erica P. Dayao1 and Mary Bernadette V. Egloso1,
Anna Theresa A. Santiago1 and Mary Ann T. Tavanlar2
1Department of Biology, College of Arts and Sciences, University of the Philippines Manila, Padre Faura St., Ermita, Manila
2National Institute of Molecular Biology and Biotechnology, University of the Philippines Los Baños, College Laguna 4031
ABSTRACT
CHICKEN FEATHER
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. Scanning Electron Microscopy (SEM) revealed colonization of Xylaria sp. strain on
the three pollutants. 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 greatest biodegrading potential for
biodegrading natural rubber. Polystyrene was effectively colonized by all strains. Chicken feathers were
colonized by the SDM and E35 but may require a longer incubation period for degradation.
INTRODUCTIION METHODOLOGY
Pollution caused by improper waste
disposal burdens third world countries like
the Philippines. It is high time to revert to
natural processes, such as biodegradation,
to solve this problem. Biodegradation is the
breakdown and utilization of waste as
source of nutrients through the enzymatic
action of microorganisms. In Mt. Makiling a
fungus named Xylaria sp. was identified
and characterized to biodegrade
polyethylene plastic. The potential ability of
these fungal strains to biodegrade other
wastes, such natural rubber, chicken
feathers and polystyrene, were tested in
this study. Results would serve as source of
preliminary information on the potential of
this fungus to biodegrade. Furthermore, the
findings would provide data on the
characteristics of these fungal strains.
RESULTS
LEGEND: my=mycelia hy=hypha sp=spores st=striation
tr=tearing c=crack r=rachis br=barb bl=barbule
p=residual polystyrene mc=mycelia
NATURAL RUBBER
All strains efficiently colonized the samples, but
only SDM and E35 showed signs of degradation
such as weakened barbs and slightly brittle
barbules which detached from their barbs.
POLYSTYRENE
Generally, the black strains specially strain E26, showed
the greatest biodegrading potential as compared to the
albino strains PNL 114, 116 and 118.
DISCUSSION
Natural rubber, polystyrene and chicken feather were utilized as co-substrates of glucose for the growth of Xylaria sp.
Mycelia closely adhered to their surface and grew into it, thereby rendering their removal very difficult. Due to this, the change
in weight was not measured. Thus, weight gain is indicative of colonization, whereas weight loss is indicative of biodegradation.
The 0.5% glucose provided in the medium initiated and sustained growth for up to 3 weeks. Beyond this period when glucose
has been depleted, Xylaria sp. had to utilize the provided substrate as an alternative carbon source.
After incubation, the set-ups were covered by mycelia. Such colonization and surface adhesion is a fundamental
prerequisite to biodegradation, although growth on a polymer surface is not adequate to deduce that carbon from this polymer
has been assimilated. Nonetheless, this colonization provides a simple, fast and clear test to evaluate the response of a
macromolecular material to biodegradation.
Gentle scraping was done after removal of fungi from the samples to avoid surface destruction and to further remove
mycelia. Hence, the colonization seen under SEM cannot be attributed to mere mycelial aggregation on the surface since
physical manipulation was done.
Most samples were covered by a slimy mucilaginous sheath or biofilm which can be easily removed. All strains tested in
this study are able to form biofilm, which is a crucial step to microbially induced biodegradation. A study hypothesized that after
Polystyrene was the most extensively colonized
adhesion, solubilizing agents are secreted by microorganisms which can make use of water-immiscible compounds.
substrate. SEM results of the albino mutants were
The denser and thicker mycelial growth and more efficient performance of the black strains SDM wild type, E26 and E35
not widely different from the black strains.
than the albino mutants (PNL 114, 116 and 118) is due to the melanin cover of their hyphal surfaces. Melanin protects their
hyphae thus making them resistant to frictional damage during penetration into the substrate. The pollutants were weakened in
resistance and durability due to mycelial penetration.
CONCLUSION ACKNOWLEDGMENT
Generally, the black strains, namely the SDM wild type, E26 and E35, have been observed to possess a higher potential to The proponents of this study would like to thank the Antibiotic Laboratory at the National Institute of
biodegrade natural rubber, chicken feather and polystyrene than the albino mutants PNL 114, 116 and 118. Among the black Molecular Biology and Biotechnology (BIOTECH) for providing the culture media, equipment and the Xylaria
variant strains, E26 has shown the greatest biodegrading potential for natural rubber, nonetheless the albino mutants also sp inocula and the National Institute of Engineering at the University of the Philippines Diliman for the
showed a potential to biodegrade this substrate. Polystyrene was effectively colonized by the three black strains SDM wild type, processing of our samples under the scanning electron microscopy (SEM).
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 Biodegrading Ability of Xylaria sp.
on Natural Rubber, Chicken Feather and Polystyrene through
Scanning Electron Microscopy
Janine Erica P. Dayao1 and Mary Bernadette V. Egloso1,
Anna Theresa A. Santiago1 and Mary Ann T. Tavanlar2
1Department of Biology, College of Arts and Sciences, University of the Philippines Manila, Padre Faura St., Ermita, Manila
2National Institute of Molecular Biology and Biotechnology, University of the Philippines Los Baños, College Laguna 4031
ABSTRACT METHODOLOGY
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. Scanning
Electron Microscopy (SEM) revealed colonization of Xylaria sp. strain on the three pollutants.
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 greatest
biodegrading potential for biodegrading natural rubber. Polystyrene was effectively colonized by
all strains. Chicken feathers were colonized by the SDM and E35 but may require a longer
incubation period for degradation.’
INTRODUCTIION
Pollution caused by improper waste disposal burdens third world countries like the Philippines. It is high
time to revert to natural processes, such as biodegradation, to solve this problem. Biodegradation is the
breakdown and utilization of waste as source of nutrients through the enzymatic action of microorganisms.
In Mt. Makiling a fungus named Xylaria sp. was identified and characterized to biodegrade polyethylene
plastic. The potential ability of these fungal strains to biodegrade other wastes, such natural rubber,
chicken feathers and polystyrene, were tested in this study. Results would serve as source of preliminary
information on the potential of this fungus to biodegrade. Furthermore, the findings would provide data on
the characteristics of these fungal strains.
RESULTS
LEGEND: my=mycelia hy=hypha sp=spores st=striation tr=tearing c=crack r=rachis br=barb
bl=barbule p=residual polystyrene mc=mycelia
NATURAL RUBBER CHICKEN FEATHER POLYSTYRENE
Generally, the black strains specially All strains efficiently colonized the samples,
but only SDM and E35 showed signs of Polystyrene was the most extensively
strain E26, showed the greatest
degradation such as weakened barbs and colonized substrate. SEM results of the
biodegrading potential as compared to
slightly brittle barbules which detached albino mutants were not widely different
the albino strains PNL 114, 116 and 118.
from their barbs. from the black strains.
DISCUSSION
Natural rubber, polystyrene and chicken feather were utilized as co-substrates of glucose for the growth of Xylaria sp. Mycelia closely adhered to their surface and grew into it, thereby rendering their removal very difficult. Due to this, the change in weight
was not measured. Thus, weight gain is indicative of colonization, whereas weight loss is indicative of biodegradation. The 0.5% glucose provided in the medium initiated (Cuevas & Manaligod, 1997; Clutario & Cuevas, 2001) and sustained growth for up to
3 weeks (M. A. Tavanlar, personal communication, March 4, 2009). Beyond this period when glucose has been depleted, Xylaria sp. had to utilize the provided substrate as an alternative carbon source. After incubation, the set-ups were covered by mycelia.
Such colonization and surface adhesion is a fundamental prerequisite to biodegradation, although growth on a polymer surface is not adequate to deduce that carbon from this polymer has been assimilated. Nonetheless, this colonization provides a simple,
fast and clear test to evaluate the response of a macromolecular material to biodegradation (Clutario & Cuevas, 2001; Motta,et al., 2007). Gentle scraping was done after removal of fungi from the samples to avoid surface destruction and to further remove
mycelia. Hence, the colonization seen under SEM cannot be attributed to mere mycelial aggregation on the surface since physical manipulation was done. Most samples were covered by a slimy mucilaginous sheath or biofilm which can be easily removed.
All strains tested in this study are able to form biofilm (M. A. Tavanlar, personal communication, March 4, 2009), which is a crucial step to microbially induced biodegradation. A study (Reddy, et al., 1982; Gutnick & Minas, 1987) hypothesized that after
adhesion, solubilizing agents are secreted by microorganisms which can make use of water-immiscible compounds (Clutario & Cuevas, 2001). The denser and thicker mycelial growth and more efficient performance of the black strains SDM wild type, E26
and E35 than the albino mutants (PNL 114, 116 and 118) is due to the melanin cover of their hyphal surfaces. Melanin protects their hyphae thus making them resistant to frictional damage during penetration into the substrate. The pollutants were
weakened in resistance and durability due to mycelial penetration.
CONCLUSION ACKNOWLEDGMENT
Generally, the black strains, namely the SDM wild type, E26 and E35, have been observed to possess a higher potential to The proponents of this study would like to thank the Antibiotic Laboratory at the National Institute of Molecular Biology and
biodegrade natural rubber, chicken feather and polystyrene than the albino mutants PNL 114, 116 and 118. Among the
Biotechnology (BIOTECH) for providing the culture media, equipment and the Xylaria sp inocula and the National Institute of
black variant strains, E26 has shown the greatest biodegrading potential for natural rubber, nonetheless the albino Engineering at the University of the Philippines Diliman for the processing of our samples under the scanning electron
mutants also showed a potential to biodegrade this substrate. Polystyrene was effectively colonized by the three black microscopy (SEM).
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.
NATURAL CHICKEN
POLYSTYRENE
RUBBER FEATHER
AGITATION w/ EtOH and SURFACE
AUTOCLAVE
STERILE WATER STERILIZATION
SDM Wild type
PNL 114
PNL 116
XYLARIA MINERAL MEDIUM
PNL 118
SP. STRAIN w/ GLUCOSE
FUNGAL INOCULATION
E26
E35
50 DAY
INCUBATION
DECANT & GENTLE CLEANING OF SUBSTRATE
SCRAPING OF MYCELIA
SCANNING ELECTRON
MICROSCOPY
Shared by: Mary Bernadette Vallesfin Egloso
Mary Bernadette Vallesfin Egloso
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