Вы находитесь на странице: 1из 9

http://informahealthcare.

com/mby
ISSN: 1040-841X (print), 1549-7828 (electronic)
Crit Rev Microbiol, Early Online: 19
! 2013 Informa Healthcare USA, Inc. DOI: 10.3109/1040841X.2013.791247

REVIEW ARTICLE

Fungal degradation of lignocellulosic residues: An aspect of improved


nutritive quality
Rakesh Kumar Sharma1 and Daljit Singh Arora2
Department of Microbiology, The Maharaja Sayajirao University of Baroda, Vadodara, India and 2Department of Microbiology, Guru Nanak Dev
University, Amritsar, India
Abstract

Keywords

Microbial degradation of lignocellulosic materials brings a variety of changes in their


bio-physicochemical properties. Lower digestibility of various agricultural residues can be
enhanced by microbial treatment. White rot fungi are the potential candidates, which can
improve the nutritional quality of lignocellulosic residues by degrading lignin and converting
complex polysaccharides into simple sugars. Changes in physical qualities of lignocellulosics
that is texture, colour and aroma have been an interesting area of study along with chemical
properties. Degradation of lignocellulose not only upgrades the quality of degraded biomass,
but helps simultaneous production of different commercial enzymes and other by products
of interest. The review is focused on fungal degradation of lignocellulosics, resultant changes in
physicochemical properties and nutritional value.

In-vitro digestibility, lignin, lignocellulose


degradation, solid state fermentation,
white rot fungi

Introduction
The availability of green fodder is a worldwide problem,
resulting in their scarcity for animal consumption. To
overcome the crisis of feed, several cereal crop residues like
wheat straw (WS), paddy straw (PS), corn stover and so on are
generally used as substitutive fodder and feed supplement.
A variety of food crops are produced around the globe, which
generate enormous agricultural residues. This lignocellulosic
biomass can serve as low cost feed stocks for production
of fuel, ethanol and other value-added commodity chemicals
(Parimala et al., 2007). As compared to forage, cereal straws
are of poor nutritive value and low digestibility because
of their higher lignin content. Lignin is a phenyl-propanoid
biopolymer, resistant to most of the microbial enzymatic
systems and non digestible by ruminants as well. The
presence of lignin and its hemicellulose binding matrix
affects the accessibility of energy rich polysaccharides
(Figure 1).
Degradation of lignin has got the potential to improve
the digestibility of such residues (Arora & Sharma, 2011).
Natural degradation of plant biomass is mainly caused by
different fungal and bacterial species. Holocellulose and
lignin are the biopolymers that are converted into low
molecular weight compounds by the enzymatic action of
these microorganisms (Bugg et al., 2011). Fungi are usually
better degraders of plant cell wall constituents because of
their extracellular enzymatic system and hyphal penetration

Address for correspondence: Daljit Singh Arora, Ph D, Department of


Microbiology, Guru Nanak Dev University, Amritsar, 143005 India.
E-mail: daljit_02@yahoo.co.in

History
Received 12 February 2013
Accepted 27 March 2013
Published online 15 July 2013

power. Many fungal species have been screened for their


potential to degrade various agricultural residues (Wan & Li,
2011). White rot fungi degrade lignin efficiently and selectively by using their ligninolytic enzyme system which mainly
comprises of laccase, lignin peroxidase and manganese
peroxidase, along with many other enzymes (Arora et al.,
2002; Arora & Sharma, 2010).
Fungal ligninolysis and break down of cellulose-hemicellulose matrix liberate simple degradable components that
can be easily utilized by rumen microflora, thus improving
the ruminant digestibility. Bioconversion of these residues
into more nutritive animal feed has fascinated many workers
to work on the problem and make it practically possible at
lab-level fermentation (Bisaria et al., 1997; Chalamcherla
et al., 2009). Solid state fermentation (SSF) of agroresidues
is an advantageous method to degrade lignin and improve
their digestibility. Fungi grown under these conditions
not only cause better ligninolysis but also improve digestibility by enhancing the accessibility of holocellulose (Okano
et al., 2006).
White rot fungi are well known producers of lignocellulolytic enzymes including lignin modifying enzymes, hemicellulases and cellulases. Degradation of holocellulose along
with lignin results into higher loss in total organic matter
(TOM), which limits the practical use of such agro residues as
feed. The degradation of useful holocellulose can be
minimized by using selective ligninolytic fungi (Tuyen
et al., 2012). Phanerocheate chrysosporium is a well known
white rot fungus and widely studied for lignin degradation
(Syed & Yadav, 2012). However, loss of TOM is very high
during the degradation of lignocellulosics, which necessitates
the need to look for selective ligninolytic organisms

20
13

Critical Reviews in Microbiology Downloaded from informahealthcare.com by 14.139.242.18 on 07/16/13


For personal use only.

R. K. Sharma & D. S. Arora

Figure 1. (A) Structure of a typical lignocellulosic residue, (B) Initial fungal attack on
lignin hemicellulose matrix, (C) Degradation
of all three polymers and fungal growth,
(D) Nutritionally upgraded lignocellulose
along with fungal proteins and simple sugars
to be used as animal feed.

Crit Rev Microbiol, Early Online: 19

(A)

Lignin-Hemicellulose
Matrix

(D)

Hemicellulose

Fungal
proteins

Cellulose
Simple
sugars

(B) Initial fungal


attack site

Critical Reviews in Microbiology Downloaded from informahealthcare.com by 14.139.242.18 on 07/16/13


For personal use only.

Lignin

(Jung et al., 1992). Selective lignin degrading organisms


possess the potential capability for improvement of digestibility of agro-residues (Arora & Gill, 2000; Vares et al., 1994).
Quality of straw depends upon the composition of its
cell wall fibers, that is neutral detergent fiber (NDF), acid
detergent fiber (ADF), water solubles, hemicellulose, cellulose, lignin, protein and ash content, where higher ADF value
and lignin content results in less digestibility (Garcia et al.,
2003). Digestibility measured by in-vitro methods gives a
close idea about the quality of feed (Goering & Van Soest,
1970). An increase in dry matter digestibility shows increased
quality of feed and less feed intake. Some physical properties
of straw, e.g. swelling capacity, moisture content, pH, etc. also
affect the digestibility and feed quality. Protein and amino
acid content of the feed are also important factors regulating
its nutritive value. Thus, different compositional analyses of
sound and delignified straw are done to know their nutritive
quality (Shrivastava et al., 2012).
In the present status report, keeping in mind the poor
nutritive quality of agricultural residues and dismal scenario
of green feed for ruminants, the fungal degradation of agroresidues using some selective ligninolytic fungi and measures
of nutritive quality along with the biochemical changes in
agroresidues is discussed. The importance of SSF as an ecofriendly system to convert abundantly available agricultural
residues in to nutritive animal feed is also being highlighted.

Evaluation of nutritional quality


According to Alves De Brito et al. (2003), evaluation of the
nutritional quality of forage requires the detailed analysis
of the composition of its cell wall. The digestibility of cell
wall is influenced by both, the contents and physical
characteristics of wall polysaccharides such as degree of
crystallinity and polymerization (Fritz et al., 1990).

(C)

Role of chemical constituents in digestibility


of plant cell wall
Composition and constituents of cell wall play an important
role to determine the digestibility. Richness in holocellulosic
components enhances the digestibility. However, lignin plays
a major role in protecting the substrate from microbial and
chemical attacks as this is the most resistant part in plant cell
wall. The high level of lignification and silicification limits
ruminal degradation of the carbohydrates and the low content
of nitrogen are the main deficiencies of rice straw, affecting
its value as feed for ruminants (Van Soest, 2006).
Ash constitutes the inorganic matter mainly containing
minerals and is also difficult for animal digestion but required
in trace amounts. As ash does not contain any energy, so it
would naturally lower the overall energy and digestibility of
the feed (Fonnesbeck et al., 1981). Minerals also act as barrier
against the attack of rumen microbes to structural carbohydrates. The physical barrier concept is best described by
making use of the rumen as the rumen being an anaerobic
environment, it is clear that access and attachment of the
microorganism to the substrate is vital if efficient cellulose hydrolysis is to be effected (Malherbe & Cloete,
2002). Fungal delignification increases the surface area of
lignocellulosics, thus providing better opportunity to the
rumen microorganisms which not only enhances the digestibility of the feed but also improves their nutritional value
(Figure 1).

In-vitro digestibility (IVD)


Digestibility measured by in-vitro methods gives a close idea
about the quality of feed (Goering & Van Soest, 1970), as this
provides a quick and precise prediction of in-vivo digestibility
in ruminants. The in-vitro procedure does a better job of

Critical Reviews in Microbiology Downloaded from informahealthcare.com by 14.139.242.18 on 07/16/13


For personal use only.

DOI: 10.3109/1040841X.2013.791247

prediction than chemical composition because it accounts


for all factors affecting digestibility, whether known or
unknown, which is not possible by present chemical methods
(Garcia et al., 2003).
The two stage in-vitro procedure developed by Tilley &
Terry (1963) is the most reliable laboratory method for
predicting the digestibility of a wide range of forages. It can
predict in-vivo digestibility with a lower error than any
chemical method and has been widely accepted throughout
the world for measuring the digestibility of feeds (Minson,
1990; Shrivastava et al., 2012). Many fungi produce cellulase,
hemicellulase and other enzymes that degrade forage carbohydrates. Jones and Hayward (1973) showed that a commercially available fungal cellulase could be used to predict
forage digestibility with an accuracy similar to that achieved
with the method of Tilley & Terry. Unfortunately, these
cellulase preparations are relatively expensive and not readily
available in less developed countries. Consequently, enzymatic methods have generally received less attention than the
procedure of Tilley & Terry.
The first stage of the Tilley & Terry (1963), technique
simulates conditions in the reticulorumen and requires an
inoculum of rumen microorganisms obtained from sheep or
cattle fitted with a rumen fistula. The use of fistulated animals
for this purpose has been criticized on ethical grounds, and
there are also many practical reasons for reducing the need
for fistulated animals in nutritional studies including (a) It
requires special surgical skills, (b) fistulated animals need
special care to ensure that the fistula is kept free of any
infection and (c) a uniform diet must be fed if the inoculum
is to have constant activity. These conditions are often
difficult or impossible to achieve in the humid tropics and
in less-developed countries (Akhter et al., 1999). It is also
difficult to handle an animal for practical use in a typical
microbiology lab.
One method of overcoming the need for rumen- fistulated
animals is to use freshly voided faeces from sheep as the
source of inoculum (El Shaer et al., 1987). Akhter et al (1999)
concluded that bovine faeces have the potential as an
alternative to rumen liquor from rumen-fistulated sheep
when estimating digestibility using the in-vitro technique.
Further, the method involving the use of enzymes like
cellulase is relatively expensive, while the one using feacal
inoculum for determining the digestibility is comparatively
cheaper and easy laboratory method and equally effective as
that of using rumen fluid (Arora & Sharma, 2009a, 2011;
Dhanoa et al., 2004; Utomo et al., 2011).

Correlation between lignin content and in-vitro


digestibility
Cell wall constituents of straw play an important role in
determining its quality as animal feed. Lignin being a
phenolic biopolymer seems to be difficult to be digested by
ruminants. Higher lignin and tannin content results into lower
digestibility of lignocellulosics and plant residues (Arigbede
et al., 2012). As evident from earlier observations, a strong
negative correlation existed between lignin content and
in-vitro digestibility of undecayed paddy straw samples,
while a strong positive correlation was observed between

Fungal degradation of lignocellulosic residues

lignin loss and in-vitro digestibility of degraded straw (Arora


& Sharma, 2009b; Sharma & Arora, 2011a). As also reported
by Cohen et al. (2002) that during selective lignin degradation, the cellulose is exposed and can be utilized by
ruminants. It strengthens the viewpoint that delignification
plays an important role in improvement of the digestibility
and feed value of straw. Several workers have been
demonstrated successful bioconversion of lignocellulosic
residues into nutritive animal feed using white rot fungi
under solid state degradation (Table 1).

Degradation of lignocellulosic residues and


in-vitro digestibility
On the basis of lignin degradation, ligninolytic fungi can be
classified into three categories (a) simultaneous, (b) nonselective and (c) selective lignin degrading fungi (Figure 2).
White rot fungi are known to attack initially on the
hemicellulose lignin matrix (Martnez et al., 2005) using
xylanase, esterase and other ligninolytic enzymes; the esterase
cleaves covalent bonds between polysaccharides and lignin
(Dong et al., 2013). Phlebia spp. degraded higher amount of
lignin selectively during SSF of lignocellulose without much
loss in cellulose (Arora & Sharma, 2009a; Sharma & Arora,
2010a). Fast growing white rot fungi P. chrysosporium grow
vigorously and degrade hemicellulose and cellulose much
faster than lignin in paddy straw while during wheat straw
(WS) degradation almost equal amount of these fibres were
degraded, while more lignin was degraded during sugarcane
baggases as compared to other fibers (Sharma & Arora,
2010b; Dong et al., 2013).
Several white rot fungi have been evaluated for their
potential to degrade lignocellulosics and their resultant effect
on digestibility. Well studied white rot fungus P. chrysosporium, non selective in lignin degradation degrades all the fibres
simultaneously which results in more holocellulose loss and
thus degraded a large amount of TOM (Chang et al., 2012).
On the other hand, Phlebia spp. degrade much lower TOM
during the fermentation process, which accounts for more
selective ligninolysis leaving behind a sufficient amount
of TOM. Thus, a reasonable amount of easily digestible
degraded biomass is available to the animal as feed (Sharma
& Arora, 2010a). For practical purposes, higher loss in TOM
severely limits the use of P. chrysosporium (Jung et al., 1992).
The degradation profile of P. chrysosporium indicates that the
higher holocellulose degradation may be due to the higher
production of polysaccharide degrading enzymes (Kerem
et al., 1992; Deshpande et al., 2009).
Cellulase and hemicellulase like enzymes break down
the complex polysaccharides into simple sugars, which
contribute to major part of water solubles. The water soluble
part provides easily digestible components which can be
used by rumen microflora and contributes to the digestibility
of straw. No direct correlation could be established between
the water soluble content, fiber degradation and digestibility
(Rolz et al., 1986). It may also be possible that the fungi
use the easily available sugars for their own growth also
(Nigam et al., 2009), which thus lead to underestimations of soluble components and may interfere with the
interpretations.

R. K. Sharma & D. S. Arora

Crit Rev Microbiol, Early Online: 19

Table 1. Comparison of some white rot fungi used for enhancement in digestibility of lignocellulosic residues.

S. No.

2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37

Ceriporiopsis subvermispora
Cyathus stercoreus
Ganoderma sp.
Candida utilis
Pleurotus ostreatus
Lentinus subnudus
Pleurotus tuber- regim
Pleurotus ostreatus
Trametes versicolor
Pleurotus sajor caju
Pleurotus pulmonarius
Pleurotus florida,
Pleurotus djamor,
Pleurotus sajor-caju
Pleurotus ostreatus
Pleurotus salmoneostramineus
Pleurotus salmoneostramineus
Lentinula edodes
Pleurotus eryngii
Ceriporiopsis subvermispora
Daedalea quercina
Hericium clathroides
Phelinus leavigatus
Inonotus andersnii
Inonotus obliquus
Inonotus dryophilus
Phlebia floridensis
Phlebia radiata
Ceriporiopsis subvermispora
Phlebia brevispora
Phlebia fascicularia
Phanerocheate chrysosporium
Phlebia brevispora
Phlebia fascicularia
Phlebia floridensis
Phlebia radiata
Ceriporiopsis subvermispora

Digestibiltiy
after
degradation
(g/kg)

Enhacement
in digestibility
(%)

Bermuda grass
stems

42

420

527

25

Wheat straw
Apple pomace

305
585

Maize husks

10
6
30
42

Wheat straw

10

299

Maize straw

40

Rice straw

60

423
423
671

Rice straw
Wheat straw
Sugarcane bagasse

50
30
64

Wheat straw

30

655
649
456
456
456
414

Wheat straw
Wheat straw

20
30

175
175

Rice straw

60

185

605
334
633
626
331
288
327
317
511
491
752
696
801
787
791
824
655
460
550
592
563
502
514
520
559
250
232
221
259
231
254
252
237
232
248
240

44
10
8
7
17
1
9
6
21
16
12
4
19
17
21
27
44
1
21
43
36
21
24
26
35
43
33
26
48
32
37
36
28
25
34
30

Lignocellulosic
substrate

% Degradation

Critical Reviews in Microbiology Downloaded from informahealthcare.com by 14.139.242.18 on 07/16/13


For personal use only.

White rot fungi

Degradation
period (days)

Initial
digestibility
(g/kg)

Symultaneous

Non selective

Selective

Types of lignin degradation


Lignin

Hemicellulose

Cellulose

Figure 2. Simultaneous lignin degradation (degradation of all three


polymers occurs with almost same rate and upto a similar quantity);
non-selective lignin degradation (degradation of lignin is lesser than
holocellulose); Selective lignin degradation (degradation of lignin is
higher than holocellulose).

284

Reference
Akin et al. 1995
Shrivastava et al. 2012
Villas-Boas et al. 2003
Jonathan et al. 2010
Shrivastava et al. 2011
Akinfemi et al. 2009
Mirzaei et al. 2007

Miki & Okano 2005


Okano et al. 2006
Jalc et al. 1997

Arora & Sharma 2009b


Arora & Sharma 2009a

Sharma & Arora 2010a

In a study on paddy straw degradation, no significant


enhancement in the in-vitro digestibility could be achieved
during initial days of straw fermentation. However, the
digestibility increases during the later period of incubation
(Sharma & Arora, 2011b) suggesting that, initially the fungi
need some easily available components for their own growth
thus limiting the availability of simple sugars for rumen
microflora. As observed by Dorado et al. (1999), composition
of the water soluble fraction correlated with the extent of
straw transformation. The initial fermentation stage (015
days) was characterized by the accumulation of water-soluble
products from straw degradation and fungal metabolism, the
concentration of which tended to stabilize in the later
stage (1660 days). In most of the cases, the water soluble
components were not liberated at a constant rate during fungal
degradation, while the fiber degradation and digestibility
increased steadily. It suggests that digestibility also depends
upon the availability of other polysaccharides as well as the
structure of polymers (Bertrand et al., 2006) that are modified
by the fungal enzymatic treatment.
Hemicelluloselignin matrix is primarily attacked by
white rot fungi moath; relatively higher amounts of lignin

Fungal degradation of lignocellulosic residues

DOI: 10.3109/1040841X.2013.791247

Glucose

Cellulose
degradation

(B) Secondary
fungal attack site

Critical Reviews in Microbiology Downloaded from informahealthcare.com by 14.139.242.18 on 07/16/13


For personal use only.

H
Hemicellulose
degradation

Lignin
degradation
on

Xylose and
other 5C
sugars
(A) Initial
fungal attack site

Small Phenolics

Figure 3. Degradation of lignocellulose by selective ligninolytic


fungi; (A) Initially, ligninases and hemicellulases trigger the degradation of lignocellulose; (B) cellulose degradation starts along with
hemicellulose.

and hemicellulose degradation as compared to cellulose.


During an earlier study (Xu et al., 2010) on corn stover
degradation by Irpex lacteus, lignin and hemicellulose were
also selectively degraded over cellulose. Degradation of
lignin and hemicellulose was correlated in a strong positive
manner (Arora & Sharma, 2009b; Tuyen et al., 2012). Even
under varied nutritional environment, lignin hemicellulose
matrix were attacked initially, indeed the rate and efficiency
of the degradation being different (Arora and Sharma, 2011;
Sharma & Arora, 2010b). Thus, hemicellulose degradation
and in-vitro digestibility may also be positively correlated
(Agosin et al., 1985). Hemicellulose and lignin showed
the largest proportionate degradation in WS after incubation
with the D. quercina, H. clathroides, P. laevigatus, and
L. obliquus, while the other fungi caused the maximum loss
in cellulose and hemicellulose contents. It suggests that the
digestion enhancement of wheat straw colonized by white
rot fungi is regulated by complex factors including the
degradation of structural carbohydrates and lignin (Jalc et al.,
1998). Figure 3 shows initial and secondary fungal attack
sites and the release of simple monomer units from the
complex polymers.
Selective ligninolysis may be characterized by a higher
positive correlation between TOM loss and lignin loss as
compared to the polysaccharide degradation. Lignin must
be a major component of degraded TOM in comparison
to hemicellulose and cellulose (Chang et al., 2012).
Ligninolysis, loss in TOM and IVD of PS is illustrated in
Figure 1. The lignin loss was relatively lesser as compared to
the loss in TOM during PS degradation by P. chrysosporium,
while in all other cases it was relatively higher than TOM loss
and clearly reflected selectivity towards lignin degradation by
all Phlebia spp (Sharma & Arora, 2010a).

Crude protein
Feed protein content is generally expressed in the form of
crude protein. Crude protein is termed crude because it is not
a direct measurement of protein but is an estimation of total
protein based on nitrogen (N  6.25 crude protein), which
assumes 16 g of N/100 g of protein in feeds. Crude protein
includes true protein and non-protein nitrogen (NPN) such as
urea nitrogen and ammonia nitrogen. The crude protein value
provides no information about amino acid composition,
intestinal digestibility of the protein, or its rumen degradability (Garcia et al., 2003; Rotz, 2004). Generally, growth of
fungal mycelium increases total protein content of the feed
(Fazaeli, 2007). Fungal degradation of straw enhanced
crude protein content (Sharma & Arora, 2010a) and in vivo
N intake by the animal (Shrivastava et al., 2012). Selection of
selective lignin degrading fungi over the cellulose and their
capability of synthesizing proteins with high nutritional
value are necessary during SSF of lignocellulosics (Zhu
et al., 2011).

Amino acids
Proteins are composed of amino acids, which are required for
the maintenance, growth, and productivity of animals. A total
of about 22 amino acids have even identified of which the
animal can synthesize about half; which are called nonessential amino acids. However, the essential amino acids
cannot be synthesized and must be provided in the diet.
Nutritionally these amino acids are important because limiting these will affect the growth and development of the
animal. Supplemental amino acids can be added to feedstuff
to increase efficiency of animal production and achieve a low
cost feed formulation. Analyzing the amino acid composition
of feedstuffs ensures that nutritionists provide a more precise
feed formulation (Rotz, 2004). SSF of lignocellulosics also
improves the available amino acid content of the residual
biomass (Chalamcherla et al., 2010; Arora & Sharma, 2011).
It is important to measure the amino acid content during
lignocellulosic degradation to be used as feed.
To measure the amino acid content during lignocellulosic
degradation to be used as feed, an instant and reliable method
is needed to analyze the samples. Amino acids produce the
typical purple-blue or yellow colour during ninhydrin reaction, which can easily be measured colorimetrically.
Ninhydrin method was introduced for quantitative determination of amino acids in the late 1940s. The method was
originally developed for chromatographic elution from amino
acid analyzer (Moore & Stein, 1948), this method has also
been adapted for determination of amino group containing
compounds in foods as well as various types of samples
(Hurst et al., 1995; Panasiuk et al., 1998). Although instrumental methods such as amino acid analyzer and HPLC are
currently used for determining compounds containing amino
group, the simple and convenient ninhydrin method still
possess several advantages because no expensive equipment
is required, and it is suitable for the routine analysis of large
numbers of samples. The method has also been used for the
quantitative analysis of amino acids in a variety of agricultural
residues (Friedman, 2004). The modifications suggested
by Sun et al. (2006) make ninhydrin method even more

R. K. Sharma & D. S. Arora

convenient, less expensive, and less time consuming for


quantification of compounds containing amino group.

Critical Reviews in Microbiology Downloaded from informahealthcare.com by 14.139.242.18 on 07/16/13


For personal use only.

Antioxidants
Vitamin E, vitamin C, carotenoids, and some trace minerals
are important antioxidant components of animal feed and
their role in animal health and immune function are
indispensable (Roeder, 1995). Free radicals are mainly
reactive oxygen species (ROS) and reactive nitrogen species
(RNS) and include not only the oxygen or nitrogen radicals,
but some non-radical reactive derivatives of oxygen and
nitrogen. These free radicals are constantly produced during
normal physiological metabolism in tissues and damage the
biologically important molecules such as DNA, proteins,
lipids, and carbohydrates (Bellomo, 1991). Under normal
conditions the deleterious effects of ROS/RNS are countered
by the bodys antioxidant defenses, which are contributed
through dietary intake of key nutrients (e.g. vitamins and trace
minerals). Antioxidants serve to stabilize these highly reactive
free radicals, thereby maintaining the structural and functional integrity of cells. Thus, along with nutritive feed the
antioxidants are very important for the immune system and
health of the animals (Chew, 1995; Weiss, 2005).
A variety of polyphenols contributes to the antioxidant
potential of feed (Gladine et al., 2007). Cereal straw contains
lignin, which are complex phenolic polymers and exhibit a
very poor solubility. This may limit their reactivity with the
radicals responsible for the oxidation and subsequently limit
their protective effect compared to that of synthetic antioxidants. As reported by Pan et al. (2006), the lignin with more
phenolic hydroxyl groups, less aliphatic hydroxyl groups, low
molecular weight, and narrow polydispersity showed high
antioxidant activity. The delignification involve the cleavage
of covalent linkages of the lignin, which results into the
formation of low-molecular weight units holding a great value
in antioxidant enhancement (Pouteau et al., 2003). Thus,
degradation of lignin enhances smaller phenolic units and
holds the potential to upgrade the quality of lignocellulosic
residue by enhancing its antioxidant property. Fungal treatment of tomato pomace and some agricultural residues to
upgrade the low-quality feed into antioxidants rich nutritional
feed (Assi & King, 2007; Lateef et al., 2008). Total phenolic
content (TPC) and antioxidant activity of WS and PS
significantly increased during fungal degradation. TPC correlates positively with antioxidant activities; indicating the
bioactive potential of phenolic compounds, which neutralize
the free radicals (Arora et al., 2011). Thus, ligninolysis of
lignocellulosic residues provides an added feature to the feed
by enhancing its antioxidant potential (Pouteau et al., 2003;
Sharma et al., 2010).

Fungal degradation of lignocellulosic residues and


associated changes in physicochemical properties
Fungal degradation of straw brings about a variety of changes
in the biological, chemical, and physical properties of the
substrates, that is change in colour, aroma, pH, and enhancement in total phenolic content. These changes may be because
of the production of intermediate compounds formed during
fungal degradation of lignin, cellulose, or hemicellulose.

Crit Rev Microbiol, Early Online: 19

Visual observation showed the change in texture (increased


degree of roughness) and colour (turned pinkish-white) during
degradation process, which might be due to (1) breakdown of
complex plant cell wall polymers, (2) fungal growth on the
substrate, and (3) simultaneous production of some metabolites/by products. Commission Internationale de leclairage
(CIE) colour measurements used for the colorimetric analysis
of degraded lignocellulose showed that yellow and red colour
was significantly higher in degraded straw (Arora et al.,
2011). This change in colour might be contributed by the
quinone like smaller phenolic compounds, which are reported
to be coloured in general and yellow or brown in specific
(Murata et al., 2002).
The change in pH may be the result of the production
of acidic compounds by the degradation of complex sugar
and lignin. A decline in pH was recorded during fungal
degradation different lignocellulosics (Dong et al., 2013).
Studies have reported the formation of a mixture of acids
including aromatic carboxylic acids during lignin degradation
e.g. benzoic acid, p-methoxybenzoic acid, veratrum acid,
i-hemipinic acid, Phenoxyacetic acid, 2-methoxyphenylacetic
acid, salycylic acid, trans-cinnamic acid and so on along with
some -keto acids, which are converted to aromatic carboxylic
acids in a second oxidation step with hydrogen peroxide
(Javor et al., 2003; Ko et al., 2009). Various pathways
describing the bacterial and fungal degradation of lignin
and its by products were well discussed during earlier review
(Bugg et al., 2011).
Similarly, the change in aroma that is having slightly
pungent smell of degraded straw indicates the presence of
aldehydes and/or acidic compounds. Several compounds
including carboxylic, aldehydes, and ketone groups were
identified as intermediate or by products before the complete
mineralization of the lignin (Maman et al., 1996). The
metabolic pathway of the fungi plays an important role in the
production of different intermediate compounds, which
depends upon their enzymatic activity, fungal strain, and
substrate. Some fungi efficiently mineralize lignin in a short
period while others take longer time and produce higher
amount of smaller/monomer phenolic units (Tuomela et al.,
2002). The studies have shown not so uniform increase in
the TPC produced, which did not show a similar rate of
enhancement as the degradation of lignin. This can be
attributed to either complete degradation of lignin to CO2 and
H2O, while in some fungi high TPC associated with low
lignin loss indicate the production of smaller phenolic units
and their further degradation to a limited extent. A positive
correlation between TPC and lignin loss showed the enhancement in smaller phenolic units during the break down of
lignin (Arora et al., 2011). The slight difference in TPC and
lignin degradation may be due to the difference in the pattern
of ligninolysis, quality, and quantity of smaller phenolic units
formed, their solubility and further transformation as well
(Yang et al., 2010).
Ash constitutes inorganic matter and is required in trace
amounts. As ash does not contain any energy, so it would
naturally lower the overall energy of the feed (Fonnesbeck
et al., 1981; Frei et al., 2011). During fungal degradation of
lignocellulosic residues the ash content seems to be increased
(Akinfemi et al., 2009). Ash content of the degraded straw

Critical Reviews in Microbiology Downloaded from informahealthcare.com by 14.139.242.18 on 07/16/13


For personal use only.

DOI: 10.3109/1040841X.2013.791247

varies depending upon the other constituents of straw as


well as the fungal species used for the degradation. The rate
and efficiency of degradation also affect the ash content
in the residual biomass. In general, organism capable of
degrading higher amount of lignocellulose leaves a higher
amount of residual ash. The lignocellulose degraded by
P. chrysosporium contains a large amount of ash (22 %),
while in the straw degraded by P. brevispora had a lower
ash content (12.6 %). This positive correlation between ash
content and TOM loss, indicates that the residual ash
constituents were not utilized by the fungi; thus the fungus
causing maximum TOM also contributes in higher ash
content. Thus, the higher ash content in degraded straw may
be a factor for its lower IVD even after degrading sufficient
amount of lignin (Sharma & Arora, 2010a). Both of these
factors (enormous loss of lignocellulosic resides and
increased ash content) must be kept in mind while choosing
the fungus to be used for processing of lignocellulosic
residues for animal feed production. In contrast, the ash
content showed a weak positive correlation with in-vitro
digestibility.

Toxicity of fermented animal feed


Different workers have used white rot fungi to upgrade the
nutritive quality of lignocellulosic residues. Apparently, no
case has been reported for the pathogenicity of these fungi
towards human and animal species. The potential for
biological hazard is low for the microbially converted feeds
so far evaluated (Banerjee et al., 2000; Sinskey & Batt, 1987).
During a recent study on fungal fermented wheat straw
(Sharma et al., 2012), different mycotoxins (aflatoxins) were
observed but the levels of the toxins in all the diets were far
below to the permissible levels (20 ppb) in the feeds meant
for immature animals (Food and Drug Administration, USA
(USFDA)) and poultry (Bureau of Indian Standards (BIS)).
Nevertheless, this hazard has to be continually evaluated by
various biological studies when a new microbial fermented
product is proposed.
Mycotoxins are secondary metabolites produced by fungal
species mainly including Aspergillus, Penicillium, Fusarium,
Alternaria, and Stachybotrys spp. During the growth of the
moulds on food and feed stuff the toxins may be produced.
A number of bioassays using cell culture techniques have
been described for the toxicological characterization of
mycotoxins. A colorimetric cell culture assay using 3-(4,5dimethylthiazolyl-2)-2,5 diphenyltetrazolium bromide (MTT)
is an important measure of cellular toxicity, firstly used by
Mosmann (1983). MTT can reflect the sensitivity of live cells
to extrinsic stimulation; therefore, it also has an important
value to assess cell viability. Earlier, to detect toxin effects in
in-vitro cell models, the (MTT) dye reduction assay has been
shown to be an effective indicator of fungal toxicity (Gilbert
& Slavik, 2004; Maenetje et al., 2008).
Some mycotoxins especially Aflatoxin B1 was reported
to be highly mutagenic in the Salmonella typhimurium
(Ames test) system (Ciegler & Bennett, 1980). Ames test
is well known for the assessment of mutagenic activity.
Feed extract against S. typhimurium was used to evaluate the
mutagenic activity of extract (Arora et al., 2011).

Fungal degradation of lignocellulosic residues

As reported by Zadrazil (2000), some fungi decompose


lignin and other substrate components, but in-vitro digestibility decreases. This may be due to toxicity for the rumen
microorganisms of substrate extracts that are used for the
determination of in-vitro digestibility. This concept is also
helpful to know about the toxicity of fermented feed towards
rumen microflora. Tests for toxicity of the organism and the
treated biomass as well as large scale setup along with animal
trials are necessary to further evaluate the potential of solid
state fungal treatment of lignocellulose (Akin et al., 1993).

Conclusions
Looking for a solution to the problem related to scarcity
of green fodder and simultaneously managing lignocellulosic
wastes seems to be a fastidious approach. A positive
correlation between ligninolysis and digestibility gives an
idea for the removal of lignin using selective lignin degrading
white rot fungi. During the degradation of complex
lignocellulosics various intermediate by products are produced, which affect the physicochemical and nutritional
quality of degraded residues. Increase in nutritive value
during fungal degradation can be summarized in following
steps (a) break down of lignin-polysaccharide binding matrix,
(b) reduction in lignin content, (c) increased surface area for
the better access of energy containing fibers, (d) conversion of
complex polysaccharides into simple sugars, (e) increased
protein content and antioxidants. The points discussed, may
be useful for analytical aspect of microbiologically treated
feed stuff.

Declaration of interest
The authors report no conflict of interest. The authors alone
are responsible for the content and writing of this article.
References
Agosin E, Monties B, Odier E. (1985). Structural changes in wheat straw
components during decay by lignin-degrading white-rot fungi in
relation to improvement of digestibility for ruminants. J Sci Food
Agric 36:92535.
Akhter S, Owen E, Theodorou MK, et al. (1999). Bovine faeces as a
source of micro-organisms for the in-vitro digestibility assay of
forages. Grass Forage Sci 54:21926.
Akin DE, Sethuraman A, Morrison WH, et al. (1993). Microbial
delignification with white-rot fungi improves forage digestibility.
Appl Environ Microbiol 59:427482.
Akin DE, Rigsby LL, Sethuraman A, et al. (1995). Alterations in
structure, chemistry, and biodegradability of grass lignocellulose
treated with the white rot fungi Ceriporiopsis subvermispora and
Cyathus stercoreus. Appl Environ Microbiol 61:15918.
Akinfemi A, Adu OA, Adebiyi OA. (2009). Use of white rot-fungi
in upgrading maize straw and, the resulting impact on chemical
composition and in-vitro digestibility. Livestock Research for Rural
Development 21:162. http://www.lrrd.org/lrrd21/10/akin21162.htm.
Alves De Brito CGF, Rodella RA, Deschamps FC. (2003). Chemical
profile of cell wall and its implications on Brachiaria brizantha and
Brachiaria humidicola digestibility. Revista Brasileira de Zootecnia
32:183544.
Arigbede OM, Tan ZL, Anele UY, et al. (2012). Effects of age and
species on agronomic performance, chemical composition and in vitro
gas production of some tropical multi-purpose tree species. J Agric Sci
150:72537.
Arora DS, Chander M, Gill PK. (2002). Involvement of lignin
peroxidase, manganese peroxidase and laccase in degradation and
selective ligninolysis of wheat straw. Int Biodeterior Biodegrad 50:
11520.

Critical Reviews in Microbiology Downloaded from informahealthcare.com by 14.139.242.18 on 07/16/13


For personal use only.

R. K. Sharma & D. S. Arora

Arora DS, Gill PK. (2000). Laccase production by some white rot fungi
under different nutritional conditions. Bioresour Technol 73:2835.
Arora DS, Sharma RK, Chandra P. (2011). Biodelignification of wheat
straw and its effect on in vitro digestibility and antioxidant properties.
Int Biodeterior Biodegrad 65:3528.
Arora DS, Sharma RK. (2009a). Comparative ligninolytic potential of
Phlebia species and their role in improvement of in vitro digestibility
of wheat straw. J Anim Feed Sci 18:15161.
Arora DS, Sharma RK. (2009b). Enhancement in in vitro digestibility
of wheat straw obtained from different geographical regions during
solid state fermentation by white rot fungi. BioRes 4:90920.
Arora DS, Sharma RK. (2010). Ligninolytic fungal laccases and
their biotechnological applications. Appl Biochem Biotechnol 160:
176088.
Arora DS, Sharma RK. (2011). Effect of different supplements on
bioprocessing of wheat straw by Phlebia brevispora: changes in its
chemical composition, in vitro digestibility and nutritional properties.
Bioresour Technol 102:808591.
Assi JA, King AJ. (2007). Assessment of selected antioxidants in tomato
pomace subsequent to treatment with the edible oyster mushroom,
Pleurotus ostreatus, under solid state fermentation. J Agric Food
Chem 55:90958.
Banerjee S, Azad SA, Vikineswary S, et al. (2000). Phototropic bacteria
as fish feed supplement. Asian Australisn J Anim Sci 13:9914.
Bellomo G. (1991). Cell damage by oxygen free radicals.
Cytotechnology 5:713.
Bertrand I, Chabbert B, Kurek B, Recous S. (2006). Can the biochemical
features and histology of wheat residues explain their decomposition
in soil? Plant and Soil 281:291307.
Bisaria R, Madan M, Vasudevan P. (1997). Utilisation of agroresidues as animal feed through bioconversion. Bioresour Technol
59:58.
Bugg TDH, Ahmad M, Hardiman EM, Rahmanpour R. (2011). Pathways
for degradation of lignin in bacteria and fungi. Nat Prod Rep 28:
188396.
Chalamcherla V, Maringanti SA, Muvva VL, et al. (2009). Use of
lignocellulolytic mutants of Pleurotus ostreatus in ruminant feed
formulations. BioRes 4:14254.
Chalamcherla VL, Singaracharya MA, Lakshmi MV. (2010).
Aminoacids profile of the lignocellulosic feed treated with
cellulase-free lignolytic mutants of Pleurotus ostreatus. BioRes 5:
25967.
Chang AJ, Fan J, Wen X. (2012). Screening of fungi capable of highly
selective degradation of lignin in rice straw. Int Biodeter Biodegrad
72:2630.
Chew BP. (1995). Antioxidant vitamins affect food animal immunity and
health. J Nut 125:1804s8s.
Ciegler A, Bennett JW. (1980). Mycotoxins and mycotoxicoses.
BioScience 30:51215.
Cohen R, Persky L, Hadar Y. (2002). Biotechnological applications and
potential of wood degrading mushrooms of the genus Pleurotus. Appl
Microbiol Biotechnol 58:58294.
Deshpande P, Nair S, Khedkar S. (2009). Water hyacinth as carbon
source for the production of cellulase by Trichoderma reesei. Appl
Biochem Biotechnol 158:55260.
Dhanoa MS, France J, Crompton LA, et al. (2004). Technical note:
a proposed method to determine the extent of degradation of a feed
in the rumen from the degradation profile obtained with the in vitro
gas production technique using faeces as the inoculum. J Anim Sci 82:
73346.
Dong XQ, Yang JS, Zhu N, et al. (2013). Sugarcane bagasse degradation
and characterization of three white-rot fungi. Bioresour Technol 131:
44351.
Dorado J, Almendros G, Camarero S, et al. (1999). Transformation
of wheat straw in the course of solid-state fermentation by four
ligninolytic basidiomycetes. Enzyme Microbial Technol 25:60512.
El Shaer HM, Omed HM, Chamberlain AG, Axford RFE. (1987). Use of
faecal organisms from sheep for the in vitro determination of
digestibility. J Agric Sci 109:2579.
Fazaeli H. (2007). Nutritive value index of treated wheat straw with
Pleurotus fungi. Biotechnol Anim Husb 23:16980.
Fonnesbeck PV, Christiansen ML, Harris LE. (1981). Linear models
for calculating digestible energy for sheep diets. J Anim Sci 52:
118396.

Crit Rev Microbiol, Early Online: 19

Frei M, Kohno Y, Wissuwa M, et al. (2011). Negative effects of


tropospheric ozone on the feed value of rice straw are mitigated by an
ozone tolerance QTL. Global Change Biol 17:231929.
Friedman M. (2004). Applications of the ninhydrin reaction for analysis
of amino acids, peptides, and proteins to agricultural and biomedical
sciences. J Agric Food Chem 52:385406.
Fritz JO, Moore KJ, Jaster EH. (1990). Digestion kinetics and cell wall
composition of Brown Midrib Sorghum  Sudan grass morphological
components. Crop Sci 30:21319.
Garcia A, Thiex N, Kalscheur K, Tjardes K. (2003). Interpreting hay &
haylage analysis. Dairy Sci Extention Extra ExEx4002 (rev).
Gilbert C, Slavik M. (2004). Determination of toxicity of Campylobacter
jejuni isolated from humans and from poultry carcasses acquired at
various stages of production. J Appl Microbiol 97:34753.
Gladine C, Morand C, Rock E, et al. (2007). Plant extracts rich in
polyphenols (PERP) are efficient antioxidants to prevent lipoperoxidation in plasma lipids from animals fed n  3 PUFA supplemented
diets. Anim Feed Sci Technol 136:28196.
Goering HK, Van Soest, PJ. (1970). Forage fiber analysis, Agriculture
handbook 379, U.S. Dept. of Agriculture.
Hurst PL, Sinclair BK, Eason JR. (1995). Amino acids interfere with the
ninhydrin assay for asparagines. Food Chem 53:4679.
Jalc D, Nerud F, Siroka P. (1998). The effectiveness of biological
treatment of wheat straw by white-rot fungi. Folia Microbiol 43:
6879.
Jalc D, Siroka P, Ceresnakova Z. (1997). Effect of six species of
white-rot basidiomycetes on the chemical composition and rumen
degradability of wheat straw. J Gen Appl Microbiol 43:1337.
Javor T, Buchberger W, Faix O. (2003). Capillary electrophoretic
determination of lignin degradation. Products obtained by permanganate oxidation. Analytica Chimica Acta 484:1817.
Jonathan SG, Akinfemi A, Adenipekun CO. (2010). Biodegradation
and in vitro digestibility of maize husks treated with edible fungi
(Pleurotus tuber-regium and Lentinus subnudus from Nigeria.
EJEAFChe 9:74250.
Jones DIH, Hayward MV. (1973). A cellulase digestion technique for
predicting the dry matter digestibility of grasses. J Sci Food Agric 24:
141926.
Jung HG, Valdez FRJ, Abad AR, et al. (1992). Effect of white rot
basidiomycetes on chemical composition and in vitro digestibility of
oat straw and alfalfa stems. J Anim Sci 70:192835.
Kerem Z, Friesen D, Hadar Y. (1992). Lignocellulose degradation during
solid state fermentation: Pleurotus ostreatus verses Phanerochaete
chrysosporium. Appl Environ Microbiol 58:11217.
Ko JJ, Shimizu Y, Ikeda K, et al. (2009). Biodegradation of high
molecular weight lignin under sulfate reducing conditions: lignin
degradability and degradation by-products. Bioresour Technol 100:
16227.
Lateef A, Oloke JK, Kana GEB, et al. (2008). Improving the quality of
agro-wastes by solid state fermentation: enhanced antioxidant
activities and nutritional qualities. World J Microbiol Biotechnol 24:
236974.
Maenetje PW, Villiers de N, Dutton MF. (2008). The use of isolated
human lymphocytes in mycotoxin cytotoxicity testing. Int J Mol Sci 9:
151526.
Malherbe S, Cloete TE. (2002). Lignocellulose biodegradation:
fundamentals and applications. Rev Environ Sci Biotechnol 1:
10514.
Maman O, Marseille F, Guillet B, et al. (1996). Separation of phenolic
aldehydes, ketones and acids from lignin degradation by capillary
zone electrophoresis. J Chromatography 755:8997.
T, Speranza M, Duenas FJR, et al. (2005). Biodegradation of
Martnez A
lignocellulosics: microbial, chemical, and enzymatic aspects of the
fungal attack of lignin. Int Microbiol 8:195204.
Minson DJ. (1990). Forage in ruminant nutrition. San Diego, CA:
Academic Press. pp. 12930.
Miki S, Okano K. (2005). In vitro digestibility of unsterilized rice straw
and wheat straw cultured with Pleurotus salmoneostramineus. Nihon
Chikusan Gakkaiho 76:40714.
Mirzaei J, Tabari M, Daroodi H. (2007). The effect of Pleurotus spp.
Fungi on chemical composition and in vitrodigestibility of rice straw.
Pak J Biol Sci 10:24604.
Moore S, Stein WH. (1948). Photometric ninhydrin method for use in the
chromatography of amino acids. J Biol Chem 176:36788.

Critical Reviews in Microbiology Downloaded from informahealthcare.com by 14.139.242.18 on 07/16/13


For personal use only.

DOI: 10.3109/1040841X.2013.791247

Mosmann T. (1983). Rapid colorimetric assay for cellular growth and


survival: application to proliferation and cytotoxicity assays. J Immun
Method 65:5563.
Murata M, Sugiura M, Sonokawa Y, et al. (2002). Properties of
chlorogenic acid quinone: relationship between browning and formation of hydrogen peroxide from a quinone solution. Biosci Biotechnol
Biochem 66:252530.
Nigam PS, Gupta N, Anthwal A. (2009). Pre-treatment of agro-industrial
residues. In: Nigam PS, Pandey A, eds. Biotechnology for agroindustrial residues utilization, 1st ed. Netherlands: Springer, 1333.
Okano K, Iida Y, Samurai M, et al. (2006). Comparison of in vitro
digestibility and chemical composition among sugarcane bagasses
treated by four white rot fungi. Anim Sci J 77:30813.
Pan X, Kadla JF, Ehara K, et al. (2006). Organosolv ethanol lignin from
hybrid poplar as a radical scavenger: relationship between lignin
structure, extraction conditions, and antioxidant activity. J Agric Food
Chem 54:580613.
Panasiuk R, Amarowicz R, Kostyra H, Sijtsma L. (1998). Determination
of a-amino nitrogen in pea protein hydrolysates: a comparison of three
analytical methods. Food Chem 62:3637.
Parimala V, Krishnani KK, Gupta BP, et al. (2007). Removal of
ammonia and nitrite from coastal water using low-cost agricultural
waste. Bull Environ Contam Toxicol 78:28893.
Pouteau C, Dole P, Cathala B, et al. (2003). Antioxidant properties of
lignin in polypropylene. Polymer Degrad Stabil 81:918.
Roeder RA. (1995). Beyond deficiency: new views of vitamins in
ruminant nutrition and health: an overview. J Nut 125:1790S1S.
Rolz C, De Leon R, De Arriola MC, De Cabrera S. (1986).
Biodelignification of lemon grass and citronella bagasse by white
rot fungi. Appl Environ Microbiol 52:60711.
Rotz CA. (2004). Management to reduce nitrogen losses in animal
production. J Anim Sci 82:E11937.
Sharma KK, Shrivastava B, Nandal P, et al. (2012). Nutritional and
toxicological assessment of white rot fermented animal feed. Ind J
Microbiol 52:18590.
Sharma RK, Arora DS. (2010a). Changes in biochemical constituents
of paddy straw during degradation by white rot fungi and its impact
on in vitro digestibility. J Appl Microbiol 109:67986.
Sharma RK, Arora DS. (2010b). Production of lignocellulolytic enzymes
and enhancement of in vitro digestibility during solid state fermentation of wheat straw by Phlebia floridensis. Bioresour Technol 101:
924853.
Sharma RK, Arora DS. (2011a). Biodegradation of paddy straw obtained
from different geographic locations by means of Phlebia spp. for
animal feed. Biodegradation 22:14352.
Sharma RK, Arora DS. (2011b). Solid state degradation of paddy
straw by Phlebia floridensis in the presence of different supplements for improving its nutritive status. Int Biodeterior Biodegrad 65:
9906.
Sharma RK, Chandra P, Arora DS. (2010). Antioxidant properties and
nutritional value of bioprocessed wheat straw by Phanerochaete
chrysosporium and Daedalea flavida. J Gen Appl Microbiol 56:
51923.

Fungal degradation of lignocellulosic residues

Shrivastava B, Thakur S, Khasa YP, et al. (2011). White-rot fungal


conversion of wheat straw to energy rich cattle feed. Biodegradation
22:82331.
Shrivastava B, Nandal P, Sharma A, et al. (2012). Solid state
bioconversion of wheat straw into digestible and nutritive ruminant
feed by Ganoderma sp. rckk02. Bioresour Technol 107:34751.
Sinskey AJ, Batt CA. (1987). Fungi as a source of protein. In: Benchat
LR, ed. Food and beverage. New York: Von Nostrand Reinhold,
43571.
Sun SW, Lin YC, Weng YM, Chen MJ. (2006). Efficiency improvements
on ninhydrin method for amino acid quantification. J Food Comp Anal
19:11217.
Syed K, Yadav JS. (2012). P450 monooxygenases (P450ome) of the
model white rot fungus Phanerochaete chrysosporium. Crit Rev
Microbiol 38:33963.
Tilley JMA, Terry, RAA. (1963). Two-stage technique for the in vitro
digestion of forage crops. J Br Grassl Soc 18:10411.
Tuomela M, Oivanen P, Hatakka A. (2002). Degradation of synthetic
14C-lignin by various white-rot fungi in soil. Soil Biol Biochem 34:
161320.
Tuyen VD, Cone JW, Baars JJP, et al. (2012). Fungal strain and
incubation period affect chemical composition and nutrient availability of wheat straw for rumen fermentation. Bioresour Technol 111:
33642.
Utomo R, Soejono M, Widyobroto BP, Sudirman. (2011). Determination
of in vitro digestibility of tropical feeds using cattle faeces as rumen
fluid alternative. Media Peternakan 34:20711.
Van Soest, PJ. (2006). Rice straw, the role of silica and treatments to
improve quality. Anim Feed Sci Technol 130:13771.
Vares T, Niemenmaa O, Hatakka A. (1994). Secretion of ligninolytic
enzymes and mineralization of 14c-ring-labelled synthetic lignin by
three Phlebia tremellosa strains. Appl Environ Microbiol 60:56957.
Villas-Boas SG, Esposito E, Mendonca MM. (2003). Bioconversion of
apple pomace into a nutritionally enriched substrate byCandida utilis
and Pleurotus ostreatus. World J Microbiol Biotechnol 19:4617.
Wan C, Li Y. (2011). Effectiveness of microbial pretreatment by
Ceriporiopsis subvermispora on different biomass feedstocks.
Bioresour Technol 102:750712.
Weiss WP. (2005). Antioxidants nutrients, cow health and milk quality.
Dairy cattle nutrition workshop. Penn State: Department of Dairy and
Animal Sciences, 1118.
Xu C, Ma F, Zhang X, Chen S. (2010). Biological pretreatment of corn
stover by Irpex lacteus for enzymatic hydrolysis. J Agric Food Chem
58:108938.
Yang X, Ma F, Zeng Y, et al. (2010). Structure alteration of lignin
in corn stover degraded by white-rot fungus Irpex lacteus CD2.
Int Biodeterior Biodegrad 64:11923.
Zadrazil F. (2000). Is the conversion of lignocellulose into feed with
white rot fungi realizable? Practical problems of scale up and
technology transfer. Mushroom Sci 15:91928.
Zhu YS, Zhang HB, Zhang YL, Huang F. (2011). Lignocellulose
degradation, enzyme production and protein enrichment by Trametes
versicolor during solid-state fermentation of corn stover. Afr J
Biotechnol 10:918292.