The Potential of Trichoderma harzianum as a Biodegradable Solution in Decomposing Low

Density and High Density Polyethylene Waste Plastics

A research proposal presented to the Faculty of College of Arts and Sciences
Cagayan State University
Carig Campus, Tuguegarao City

In partial fulfillment of the requirements of the
Degree of Bachelor of Science in Biology

Lina Riza N. Montero


Prof. Osmond Narag
Research Instructor



Chapter I
Introduction
Plastics are ubiquitous in the modern world. Polyethylene is the world's most
common plastic. It finds innumerable applications in everything from bottles and jugs to
shopping bags and children's toys. It owes its surprising versatility to its properties and molecular
structure. There are different kinds of polyethylene available; two of the most common are high-
density polyethylene or HDPE and low-density polyethylene or LDPE.
LDPE was the first PE to be developed. It has low-density levels and only a small amount
of branching. It is very flexible and easy to clean. It is often used to make plastic film wrap and
plastic bags. Additionally, it is used to make plastic items that need to be molded, such as plastic
bottles used in labs. While HDPE has higher density levels; it is characterized by a linear
structure consisting of no branching. That makes HDPE stronger and more resistant to
chemicals. It is most commonly used for items requiring blow molding techniques, such as toys,
automobile parts and bottles. It is also used to create cutting boards since it meets FDA food
service standards.
Although most of these plastics can be recycled, much of it ends up in our trash cans.
There are concerns in the environmental industry that, since these plastics do not break down
easily in a landfill, there will be many future negative effects due to our overuse of these
products. According to the American Chemistry Council's Resin Production and Sales Stats
report, released in November 2009, HDPE is the most common plastic and LDPE is the fifth
most common plastic produced in the U.S. The slow rate at which plastic degrades vs. the
amount produced makes the disposal of it an environmental issue. To overcome this ever-
increasing serious problem, decomposition of waste plastics seems to be a fruitful solution.
Certain bacterial strains had been discovered to degrade these plastics, however, seeing
that there are various fungal strains that have the potential to be involved in the process of
decomposition that can contribute to the adamant pollution and equilibrium in nature. This study
will provide information regarding the potency of Trichoderma particularly the T. harzianum as
a natural and valuable decomposing agent in non- biodegradable matter in order to reduce the
utilization of chemicals capable of decomposition.
Trichoderma is a genus of fungi that is present in all soils, decaying wood and vegetable
matter. Their dominance in soil may be attributed to their diverse metabolic capability and
aggressive competitive nature (Lewis and Papavizas, 1991). These characteristics make them
significant decomposers of woody and herbaceous material and are also necrotrophic against
other decomposers. Many species in this genus can be characterized as opportunistic a virulent
plant symbionts, the ability of several Trichoderma species to form mutualistic endophytic
relationships with several plant species. One of which species is the Trichoderma harzianum, the
most frequent Trichoderma species cultivated from soil worldwide. That displays a remarkable
diversity of lifestyles ranging from saprotrophy in free soil and dead wood, in rhizosphere and on
dead fungal biomass to biotrophy in necrotrophic mycoparasisitic attacks of other fungi and
endophytic associations with plants (http://genome.jgi.doe.gov). In addition to, it is reported that
T. harzianum may have the capability of degrading organochlorine pesticides such as DDT,
dieldrin, endosulfan, pentachloronitrobenzene, pentachlorophenol and hence has potential
applications for bioremediation (Kelley, 1976). These pesticides contain petroleum oils that are
refined from crude oil which is the main ingredient of plastic.
Objectives of the Study
Generally, this study aims to understand the microphysical potency of T. harzianum as an
effective biodegradable solution in decomposing waste plastics. It specifically aims to:
To describe the potential of T. harzianum as potential decomposer,
To observe the physical manifestations of exposing LPDE and HPDE plastics to
T. harzianum under experimental condition,
To determine the efficiency of T. harzianum as a decomposer,
To determine the rate of decomposition made by T. harzianum to LPDE and
HPDE plastics.

Scope and Delimitation of the Study
This study is intended only for the determination and provision of Trichoderma
specifically the Trichoderma harzianum as potential and effective decomposer of non-
biodegradable plastics (HDPE and LDPE).

Time and Place of the Study
Cagayan State University, Carig Campus, Tuguegarao City, DOST Lab from September
to November 2014.





Definition of terms
Composting is the decomposition of plant remains and other materials to reduce the
volume of garbage needlessly sent to landfills for disposal.
Decomposer organisms that break down unused dead material that carry out the natural
process of decomposition.
Fungi any member of a large group of eukaryotic organisms that includes
microorganisms such as yeasts and molds, as well as the more familiar mushrooms.
High-density polyethylene (HDPE) or polyethylene high-density (PEHD) is a
polyethylene thermoplastic made from petroleum.
Low-density polyethylene (LDPE) is a thermoplastic made from the monomer ethylene
Mycoparasitic is a parasitic fungus whose host is another fungus.
Necrotrophic a parasite that kills its host, then feeds on the dead matter.
Polyethylene is the most common plastic the polymer that makes grocery bags
Potato Dextrose Agar is a nonselective medium for the cultivation of yeasts and molds.
Trichoderma a genus of fungi that is present in all soils, where they are the most
prevalent culturable fungi. Many species in this genus can be characterized as opportunistic
avirulent plant symbionts.
T. harzianum are the most frequent Trichoderma species cultivated from soil worldwide
that may have the potential to degrade plastics.






Chapter II

Review of Related Literature
The Nature and Effect of Plastics
Plastics have become a necessary commodity in todays world. Everyone knowingly or
unknowingly uses plastic substances. Karki (2008) discusses that plastic is used not only for
making plastic bags but also for producing products that cover parts of vehicles that need to be
protected. The word plastic comes from the Greek word plastikos, which means, able to be
molded into different shapes (Joel FR., 1995). The plastics we use are made from, inorganic and
organic raw material such as carbon, silicon, nitrogen, oxygen, chloride and hydrogen. Basic
material used for making plastic are extracted from coal, oil and natural gas. Plastics are defined
as the polymers which become mobile on heating and thus can be cast into moulds. Plastics are
nonmetallic mouldable compounds and the materials made from them, can be pushed into almost
any desirable shape and then retain that shape (Seymour RB., 1989) Commodity plastics are used
in packaging, disposable diaper backing, fishing nets and agricultural film. They include
polymers such as polyethylene, polypropylene, polystyrene, polyvinylchloride, polyurethane,
polyethyleneterepthalate, nylon (Shah, 2007). Improperly disposed plastic materials are a
significant source of environmental pollution, potentially harming life. The plastic sheets or bags
do not allow water and air to go into earth which causes infertility of soil, preventing degradation
of other normal substances, depletion of underground water source and danger to animal life. In
seas also plastic rubbish from ropes and nets to the plastic bands from beer packs chokes and
entangles marine mammals (Cooper and Vaughan, 1967). According to the municipal
administrators carry bags are the main cause of blocked drains and thus municipal wastes cannot
be incinerated leading to accumulated garage, sludge, junk (Roff and Scott, 1971). On this living
planet, a biosphere, plastic is a raging parasite that devours and pollutes everything
(http://www.mnn.com/greentech/research-innovations/blogs/boydiscovers-microbe-that-eats-
plastic).
Fungi as a Decomposer
Fungi are any member of a large group of eukaryotic organisms that includes
microorganisms such as yeasts and molds, as well as the more familiar mushrooms. Abundant
worldwide, most fungi are inconspicuous because of the small size of their structures, and their
cryptic lifestyles in soil, on dead matter, and as symbionts of plants, animals, or other fungi.
Fungi perform an essential role in the decomposition of organic matter and have fundamental
roles in nutrient cycling and exchange (http://en.wikipedia.org). Most fungi are decomposers.
Fungi break down, or decompose, the complex carbon compounds that are part of living matter.
They absorb nutrients and leave behind simpler compounds. Fungi are heterotrophs. They get
their energy from living or once living matter. They, along with bacteria, decompose the bodies
of dead plants and animals. They also decompose materials left behind by organisms, such as
fallen leaves, shed skin, and animal droppings. One interesting application of a mold is the use of
the fungus Trichoderma. This mold grows in soil. The digestive chemicals it produces are used
to give blue jeans a stonewashed look
(www.d123.org/olhms/ebarlos/documents/Fungi2.4C3.pdf ). In addition to, several reports
appeared that fungi dont just decompose organic matter but also degrade inorganic ones like
Polyurethanes (PU) that are present in many aspects of modern life. They represent a class of
polymers that have found a widespread use in the medical, automotive and industrial fields.
Studies revealed that polyester-type PUs is more susceptible to fungal attack than other forms
(Kaplan et al., 1968).
Studies Related to the Decomposition of Waste plastics using Fungi
When UV rays strike plastic, they break the bonds holding the long molecular chain
together. Over time, this can turn a big piece of plastic into lots of little pieces. However, plastic
buried in a landfill rarely sees the light of the day, but in the ocean which is where a lot of plastic
wastes end up. Plastics are bathed in as much light as water. In 2009, researchers from Nihon
University in Chiba Japan, found that in warm ocean water can degrade in as little as a year. This
doesnt sound so bad until you realize those small bits of plastic are toxic chemicals such as
bisphenol A (BPA) and PS oligomer (Harris and William, 2009). In the early 1980s the research
on degradability of plastics began. Past research has isolated and identified the use of
Trichoderma spp. for bioconversion of solid wastes (kitchen waste, humus, compost and soil). In
the study, a total of 135 isolates of Trichoderma were isolated. These 135 isolates were divided
into 5 aggregate groups. Representative isolates from each group were sent for identification. It
is then concluded that the most frequently isolated species was T. harzianum and was identified
as an effective agent for solid waste conversion using spore suspension. A same study targeted
the potential of fungal isolates as a potential bioconversion agent of municipal solid waste.
Samples of fungal strains were collected from different waste disposal site. Overall among the 5
fungal strains used in the study Tricoderma spp. were the most effective strain for the solid waste
decomposition. Bari (2007), reported that T. harzianum was the most effective strain for solid
waste decomposition and showed the highest weight loss (31.8%) when the culture disc approach
was used but present results are partially in accordance with findings of Zheng and Shetty and
Martin and Dale. These plastics differ in degradation rate, application, and price. In one
development, plastics' inertness and resistance to microbial attack was reduced by incorporating
starch and later prooxidants (transition metals and oil) (Griffin, 1973). Three types of
degradation of polyethylene in these degradable starch-polyethylene polymers can occur by
different molecular mechanisms: chemical degradation, photodegradation and biological
degradation. Chemical degradation occurs when the prooxidants catalyze the formation of free
radicals in polyethylene, which react with molecular oxygen to attack the polyethylene matrix
(Johnson et al.). Heat and oxygen accelerate this chain scission of the polyethylene.
Photodegradation also occurs within the polyethylene matrix whereby UV light catalyzes the
autoxidation and generation of free radicals (David et al., 1992). Biological degradation of these
polyethylene films has been reported in pureculture studies with various microorganisms such as
Streptomyces sp. (Lee et al., 1991), Phanerochaete sp.(Ali et al., 2009), Penicillium, Fusarium,,
Alternaria, Spicaria spp., Aspergillus (Ibrahim et al., 2011), Aureobasidium, Poecilomyces
(Mehdi et al., 2010) after chemical degradation was initiated and with their corresponding
extracellular enzymes.
The Mechanism of Trichoderma spp.
Trichoderma spp. are free- living that are highly interactive in root, soil and foliar
environments. It has been known for many years that they produce a wide range of antibiotic
substances and that they parasitize other fungi. The first description of a fungus named
Trichoderma dates back to 17941 (Persoon, 1794), and in 1865 a link to the state of a Hypocrea
species was suggested (Tulasne, 1865). But the different species assigned to the genus
Trichoderma hypocrea were difficult to distinguish morphologically. It was even proposed to
reduce taxonomy to only a single species, Trichoderma viride. Hence, it took until 1969 that
development of a concept for identification was initiated (Rifai, 1969; Samuels, 2006).
Thereafter, numerous new species of Trichodermal hypocrea were discovered, and by 2006 the
genus already comprised more than 100 phylogenetically defined species (Druzhininae et al.,
2006). Trichoderma spp. are ubiquitous colonizers of cellulosic materials and are often found
wherever decaying plant material is available (Kubicek et al. 2008), as well as in the rhizosphere
of plants where they can induce systemic resistance against pathogens (Harman, 2000). The
search for potent biomass regarding enzymes and organisms also led to isolation of these fungi
from unexpected sources such as cockroaches (Yoder et al. 2008), marine mussels and shellfish
(Sallenave et al. 1999). Trichoderma spp. are characterized by rapid growth, mostly bright green
conidia and a repetitively branched condiophore structure (Gams and Bissett, 1998). The sexual
stage when found is within the Ascomycetes in the genus Hypocrea. Trichoderma spp. possesses
innate resistance to most agricultural chemicals, including fungicides, although individual strains
differ in their resistance. Some lines have been selected or modified to be resistant to specific
chemicals. Composting is the most suitable option among the wastes management strategies with
economic and environmental profits since this process reduces the bulk volume of organic
materials, eliminates the risk of spreading of pathogens, weed seeds or parasites associated with
direct land application of manure and leads to final stabilized products which can improve and
sustain soil fertility. However, composting of lignocellulosic EFB takes a longer period of time
which is considered as the most blocking stump of this eco-friendly disposal technique (Chen et
al., 1992). Trichoderma spp. are widely known as a lignocellulose decomposer because they are
filamentous and have the ability to produce profilic spores which can invade substrates quickly
(Tengerdy and Szakacs, 2003). Various studies have shown that composting of lignocellulosic
materials preinoculated with potential Trichoderma spp. can reduce the time of biodegradation
(Mohammed et al., 2012). However, for an economically competitive process an increase in
efficiency of more than 40 fold would be necessary, which is a formidable challenge for research
with Trichoderma. Besides these major applications of Trichoderma spp. the fields of green and
white biotechnology become increasingly important for environmentally safe production of
enzymes and antibiotics. The extensive studies on diverse physiological traits available and still
progressing for Trichoderma make these fungi versatile model organisms for research on both
industrial fermentations as well as natural phenomena.
Trichoderma harzainum
The filamentous fungus Trichoderma harzianum is the genetically distinct temperate
agamospecies belonging to the group of closely related (cryptic), albeit diverse, species of the
Harzianum clade of Trichoderma (teleomorph Hypocrea, Ascomycota, Dikarya). In the broad
taxonomic sense these fungi (T. harzianum) are the most frequent Trichoderma species
cultivated from soil worldwide. They display a remarkable diversity of lifestyles ranging from
saprotrophy in free soil and dead wood, in rhizosphere and on dead fungal biomass to biotrophy
in necrotrophic mycoparasisitic attacks of other fungi and endophytic associations with plants.
Because of its mycotrophyc ability T. harzianum has often been set equal to Trichoderma-based
biocontrol agents in general, as it is the principal component in several commercial biofungicide
formulations. It is used for foliar application, seed and soil treatments for suppression of various
diseases causing by such pathogens as Botrytis, Fusarium and Penicillium sp. Although T.
harzianum is not a causative agent of the green mold disease on mushroom farms it is frequently
isolated from infected cultures of Agaricus and Pleurotus and respective substrata. Interestingly,
the causative agents of the mushroom green mold diseases (T. aggressivum, T. pleurotum and T.
pleuroticola, respectively) also belong to the Harzianum clade, i.e. are closely related to T.
harzianum. Similar to T. virens (teleomorph Hypocrea virens), a rhizosphere-competent T.
harzianum may not only grow on plant roots, but its hyphae penetrate root epidermis
(endophytism), which enhances plant growth and immune system. Some molecular mechanisms
of Trichoderma mycotrophy and interactions with plants - such as the role and regulation of
formation of cell wall hydrolytic enzymes and antagonistic secondary metabolites - have been
intensively investigated in T. harzianum. It is reported that T. harzianum is capable of degrading
organochlorine pesticides such as DDT, dieldrin, endosulfan, pentachloronitrobenzene,
pentachlorophenol and hence has potential applications for bioremediation (Kelley, 1976). As a
mycoparasitic and antagonistic fungus T. harzianum is suggested to be a powerful environmental
opportunist, which is able to interplay in communities of invasive Trichoderma spp. in various
disturbed ecosystems and thus replace or suppress the local mycofauna. Hence the genome
sequence of such an outstanding opportunistic fungus as T. harzianum is expected to provide a
platform to identify genetic resource to be used in pest control, development of biofungicides,
improvement of plant health, decomposition of plastics and environmental monitoring
(http://genome.jgi.doe.gov/).











Chapter III
Materials and Methods

Materials
The equipment, supplies and materials that will be used in this study are the following:
cultured stock of T. harzianum (culture pellets & spore suspension), petri dish, LDPE and HDPE
plastics, Erlenmayer flask (150 mL), 2% Tween 20, centrifuge, test tubes, scissors, incubator,
autoclave, PDA (potato dextrose agar), Potato Dextrose Broth, distilled water and record
notebook.

Methods
Treatments
The following treatments that will be used to conduct the study are:
T
1
Control1 LDPE (no inoculation)
T
2
Control 2 HDPE (no inoculation)
T
3
50g of Low Density Polyethylene (LDPE) cut into 2-3mm with spore suspension of T.
harzianum
T
4
50g of Low Density Polyethylene (LDPE) cut into 2-3mm with culture pellet of T.
harzianum
T
5
50g of High Density Polyethylene (HDPE) cut into 2-3mm with spore suspension of T.
harzianum
T
6
50g of High Density Polyethylene (HDPE) cut into 2-3mm with culture pellet of T.
harzianum
Procedures
A. Obtaining of Subculture and Treatments
T. harzianum, will be obtained from the culture stock of the University of the Philippines
Los Banos, Campus Laboratory. The fungus will be maintained in Petri dishes of PDA (potato
dextrose agar). After inoculating from the original slant culture stock, the petri dishes will be
incubated at 28
0
C for 4-7 days and subsequently stored at 5
0
C. Spore suspension will be obtained
by washing the Petri dish cultures with a sterile aqueous solution of 2% Tween 20. The resulting
suspension will be centrifuged (2800g, 5 min). Fungal pellets will be obtained from the
germination of spores that are suspended in shake flasks in the preliminary cultivation stage.
B. Preparation of waste plastics for decomposition
LDPE and HDPE plastics will be collected from the garbage center of the CSU, Carig
Campus. Aseptic condition will be maintained as far as possible during the collection. The
samples will be cut into 2-3 mm pieces and 50 g of each will be aliquotted into a 150 mL
Erlenmayer flask, which will then be sealed with a cotton plug. The Erlenmayer flask containing
waste plastic will then be autoclaved at 121 C for 15 min.
C. Liquid shaking culture test method
A 50g plastic bag cut into 2-3mm will be added to Erlenmeyer flask (150 mL)containing
10 mL Potato Dextrose Broth, sterilized by autoclave at 121 C for 20 min. The test isolate (3
culture pellets/50g) will be added to the test medium, properly stoppered, and incubated at 30 C
with reciprocal shaking (120 oscillations/min). The degradation of the LDPE and HDPE plastics
will be monitored by measuring the weight of the plastics before and after incubation. At the end
of the incubation period, the pieces of plastics will be taken out and washed several times with
distilled water. Next, they will be dried overnight at 80C and the changes will be observed.

D. Petri dish test method
Potato Dextrose Agar will be prepared by adding 15 g of agar to 1 L of basal medium, followed
by autoclaving for 20 min and pouring it into petri dishes. Sterilized pieces of plastic bag will be
overlaid on the medium surface, and the test isolate (1.0 mL of spore suspension) will be added
over the plastic bag pieces. At the end of incubation period, the pieces of plastic bag will be
taken out and will be washed several times with distilled water. They will be next dried over
night at 80 C and the changes will be observed.
E. Data Collection Procedures
The data will be collected based on the observations that will be made during the
experiment like changes in color, odor, volume loss and weight loss of waste plastic bag. It will
be observed at 10-day intervals up to 60 days.
For measurement of volume loss (%), the following formula will be used:
Volume loss (%) = V-V1/V 100 where V is initial volume, V1 is final volume.
Weight loss (%) of plastic bag will be calculated employing the formula:
Weight loss (%) = W-W1/W 100 where W is initial weight, W1 is final weight.
F. Statistical Tools and Analysis
The experiment will be conducted using a complete randomized design (CRD) with 3
treatments replicated 3 times. Correlations will be calculated between the samples physio-
chemical characteristics and cfus. Results of all analyses will be judged for significance at the
5% level. The data will be statistically analyzed with the help of the computer package MSTAT-
C.


Literature Cited
Andleeb, Naima Atique, Pir Bux Ghumro, Safia Ahmed and Abdul Hameed. Studies on
Biodegradation of Cellulose Blended Polyvinyl Chloride Films, International Journal Of
Agriculture and Biology. Aug. 2009: 09175.
Amer Ali Shah. Role of microorganisms in the biodegradation of plastics. Department of
Microbiology Islamabad, 1 May 2007.
Bari M.A., Begum, M.F., Sarker, K.K., Rahma, M.A., Kabir, A.H., Alam, M.F. Mode of action
of Trichoderma spp. on organic solid waste for bioconversion. Plant Environ. Develop., 2007,1:
61-66.
Chen, Y., Inbar, Y., Hadar, Y. Composted residues reduce peat and pesticide use. Biocycle. June
1992, 4851.
Cooper W. and Vaughan G. Recent Developments in Polymerization of Conjugated dienes.
Progress in Polymer Science. 14 May 1967: 91-160.
David, C., Trojan M. and Daro A. Photodegradation of polyethylene: comparison of various
photoinitiators in natural weathering conditions. Polymer Degradation Stability. Sept. 1992;
37:233- 245.
Druzhinina, I.S., Kopchinskiy, A.G., Komon, M., Bissett, J., Szakacs, G., Kubicek, C.P. An
oligonucleotide barcode for species identification in Trichoderma and Hypocrea. Fungal Genet
Biol PubMed. Mar. 2005;24:813-828. <www.ncbi.nlm.nih.gov/pubmed/16154784>.
Gams W., Bissett J. Morphology and identification of Trichoderma. In: Harmann GE, Kubicek
CP, editors. Trichoderma and Gliocladium. London: Taylor and Francis; 1998, pp. 3-34.
Griffin, G.J.L. Biodegradable fillers in thermoplastics. American Chemical Society Division of
Polymer Chemistry. 1 June 1973;33:88-92.
Ibrahim N Ibrahim, Anwar Maraqa, Khalid M Hameed, Ismail M Saadoun and Hamzah M
Joel F.R. Polymer science and technology: Introduction to polymer science. May, 1995: 4-9.
Maswadeh. Assessment of potential plastic degrading fungi in Jordanian habitats. 13 Oct. 2011.
<journals.tubitak.gov.tr/biology/issues/biy-11.../biy-35-5-3-0901-9.pdf >.
Kaplan, A. M., R. T. Darby, M. Greenberger, and M. R. Rogers. Microbiological deterioration of
polyutherene systems. Journal of Industrial Microbiology and Biotechnology Oct. 1998; 9:201-
217.
Kelley, W.D. Evaluation of Trichoderma harzianum impregnated clay granules as a biocontrol
for damping off of pine seedlings caused by Phytophyhora cinnamomi. Phytopathology Jan.
1976; 66: 1023-27.
Lee, B., Pometto A.L., Fratzke A. and Bailey T.B. Application on microbiology 1991.
Masayuki, Shimao. Biodegradation of plastics. Current Opinion in Biotechnology.
2001;12:242247.
Martin S.B., Dale J.L. Biodegradation of turf thatch with wood decay fungi. Phytopathology,
Jan. 1980; 70: 297-371.
Mehdi, Borghei, Abdolreza Karbassi, Shahrzad Khoramnejadian, Abdolrasoul Oromiehie and
Amir hossein Javid. Microbial biodegradable potato starch based low density polyethylene.
African Journal of Biotechnology. 9 Mar. 2010; 9:4075-4080.
Mohammad, N., Alam M.Z., Kabbashi N.A., Ahsan A. Effective composting of oil palm
industrial waste by filamentous fungi: A review. Resources Conservation and Recycling
Aug. 2012; 58: 69 78.
Person, C.H. Disposita methodica fungorum. Neues Magazin fr die Botanik. Jan. 1794; 1:81-
128.
Rifai, M.A. A revision of the genus Trichoderma. Mycological papers no. 116. Commonwealth
Mycological Institute, Kew, Surrey, England. 1969.
Roff, W.J. and Scott J.R. Fibres, film, plastics and rubbers, butterworths. London , May
1971:66-71.
Salma S. and Gunarto. Trichoderma activity on cellulose degradation. Penelitian Pertanian
Tanaman Pangan. 1996, 15:43
Sallenave C, Pouchus YF, Bardouil M, Lassus P, Roquebert MF, Verbist JF. Bioaccumulation of
mytoxins by selfish: contamination of mussels by metabolites of a Trichoderma pseudokoningii
SMF2. FEMS Microbiol Lett. Oct. 2009;299: 135-142.
Samuels, G.J. Trichoderma: A review of biology and systematics of the genus. Mycological
Research. 1996: 923-935.
Scott, G. Photo-biodegradable plastics. Their role in the protection of the environment, Polym
Degrad Stability. 1990;29:135- 154.
Seymour R.B. Polymer Science Before and After 1899: Notable developments during the life
time of Mautis Dekker. J Macromol Sci chem. 1989;26:1023-1032.
Tengerdy, R.P., Szakacs G. Biodegradation of lignocellulose in solid substrate fermentation.
Biochem Eng. 2003: 169-179.
Thomas-Hope, H.E. Solid waste management: Critical issues for developing countries.
University of the West Indies Press, Kingston, Jamaica: Canoe, 1998.
Yoder, J.A., Glenn B.D., Benoit J.B., Zettler L.W. The giant Madagascar hissing-cockroach
(Gromphadorhina portentosa) as a source of antagonistic moulds: concerns arising from its use in
a public setting. Mycoses. 2008. 51: 95-98. . (PubMed)
Zheng, Z., Shetty K. Cranberry processing waste for solid fungal inoculants production.
Proc.Biochem., 1998, 33: 323-329.