WJM
World Journal of Microbiology
Vol. 2(2), pp. 026-032, September, 2015. © www.premierpublishers.org, ISSN: 2141-5032x
Research Article
Studies on cellulose degrading microorganisms
associated with rumen of ruminant animals
Sambo Sadiya1* and Salau A Ibrahim2
1*,2
Department of Biology, Shehu Shagari College of Education, Sokoto, Nigeria
Studies on cellulose degrading microorganisms associated with rumen of ruminants was
carried out from ruminants (ram, cow, and goat), through culture, microscopic identification,
Biochemical test and cellulose degrading methods. In the rumen content of ram four bacteria
were isolated Bacteriodes and Staphylococcus had the highest percentage (33.3%) each while
Veillonella and Bacillus had 16.6% each. Seven bacteria were isolated from cow with
Streptococcus having (22.2%) Staphylococcus (22.2%), while Bacteroides, Yersinia,
Peptococcus, Nesseria and Bacillus had equal distribution. Goat had eight bacteria including
Bacteroides, Clostridium, Yersinia, Staphylococcus, Homofermentative Lactobacillus Alcaligens
and Bacillus all of which had equal distribution. Bacteroides and Bacillus are common in all
rumens, with Bacteroides, being more prevalent in the ram. study revealed that ruminants
harbors various organisms that are active cellulose degraders, out of which Bacteroides specie
grow best on cellulose agar. For fungi, Aspergillus flavus and Aspergillus fumigatus highly
degrades cellulose, Scopulariopsis candida degrades minimally. The study revealed that
ruminants harbors various organisms that are active cellulose degraders, out of which
Bacteroides specie grow best on cellulose agar. Therefore, Rumen should be used as a site for
isolation of micro organisms capable of cellulose hydrolysis in order to reduce the coast of
purchasing commercial enzymes
Key words: Ruminant, degrading, bacteria, fungi and cellulose.
INTRODUCTION
A ruminant from a physiological point of view is any
artiodactyl mammal that digests it's food in two steps, first
by eating the raw material and regurgitating a semidigested form known as cud from within their first
stomach, known as the rumen. The process of chewing
the cud to break down the plant matter and stimulate
digestion is called ruminating (Vankessel et al., 2002).
are present in contents this consist of bacteria, protozoa
and fungi. Bacteria require warm moist free range of 38
to 42 degree Celsius. In the rumen digestion of cellulose
and other polysaccharides occur through as hollow
muscular organ which develop anatomically in size,
structure and microbial activity as the ruminant diet is
changed.
By physiology, the rumen is regarded as
fermentation vat that can hold 160 to 240 liters of material
providing
an
anaerobic
environment,
constant
temperature and pH, and it is the site of microbial activity.
An estimated 150 billion microorganisms per teaspoon
*Corresponding author: Department of Biology, Shehu
Shagari College of Education, Sokoto, Nigeria. Email:
[email protected], Tel: +2348064535420
Studies on cellulose degrading microorganisms associated with rumen of ruminant animals
Sambo and Salau
026
For example in calf the rumen gets enlarged when the
diet is changed from liquid milk to diary feed or silage.
Unique features of the rumen as site of cellulose
digestion are its relatively large size and it's position in
the alimentary tract as the organ where ingested food
goes (Lynd et al, 2002). Field et al., (2000) added that the
rumen is an anaerobic ecosystem colonized by an
extremely dense microflora and microfauna, these
populations are composed of a grand variety of bacterial
and protozoa species. The different hydrolytic,
fermentative and methanogenic activities of these
populations ensure the efficient degradation of cell wall
constituents in forages. The omasum is sometimes
referred to as the "many piles" because of its many layers
of muscular tissue. In the omasum, the particle size of
digesta is reduced and any excess water is removed
before the digesta enters the abomasums, the omasum
can contain up to 16 liters of digesta. In the abomasums
or "truestomach" acids and enzymes further digest the
ruminant's digesta. It is the first true granular portion of
the gastromtestinal tract where the stomach walls secrete
enzymes. This stomach compartment can hold
approximately 20 liters of material. The hydrochloric acid
production which converts pepsinogen to pepsin, which
breaks down protein to shorter molecular chains
compounds such as peptides and amino acids for further
digestion and absorption in the small intestine. As digesta
passes through the small intestine the pH increase at
relatively slow rate. Bile salts which are synthesized by
the liver from chlolesterol, and in maintaining this alkaline
pH in the small intestine (Ellis et al, 2005).
Bacteria are the numerous of the rumen
inhabitants and numbers up to 100 million cells per
milliliter of fluid. There are 60-100 species regarded as
normal flora inhabitants and although each species can
only undertake a few specific tasks, the bacteria as a
whole are capable of degrading all the constituents of a
plants based diet (Bhat, 2000). There is overlap between
functions of different species; some species are highly
versatile in function, while others are more specialized.
End products of the action of some species are further
metabolized by other species and are therefore
intermediates and product of the mixed population
(Morgavi et al, 2001).
Fungi are well known agents of decomposition of
organic matter in general, and of cellulosic substrate in
particular Wang et al., (2001). The primitive group of
fungi, the anaerobic Chytridomyces are well for the ability
to degrade cellulose in the gastro intestinal tracts of
ruminant animals, although taxonomy of this group
remain
controversial,
members
of
the
order
Neocallimastigates have been classified based on the
morphology of their motile zoosphores and vegetative
thalli, they include the Neocallimastigates, piromyces and
Caeccomyces (Lynd et al., 2002). Cellulolytic capability is
also well represented among the remaining subdivisions
of aerobic fungi, within the 700 species of zygomycetes
only certain members of the genus. Mucor have been
shown to posse significant cellulolytic activity. Other
division such as Ascomycetes member of the general e.g
Bulgarria, Chactomium and Helotium; members of the
division Deutromycetes e.g those of the genera Poria,
Aspergillus, Cladosporium, Fusarium, Greotichum,
Penicillium and Trichoderma have cellulolytic enzymes
(Masoni et al., 2007).
MATERIALS AND METHODS
Sample collection
A portion of the rumen of each animal (ram, cow,
and goat) were cut with content of digested food
immediately they were slaughtered and taken to the
microbiology research laboratory Usmanu Danfodio
University Sokoto for further analysis. Nutrient agar
saboraud dextrose agar (SDA), cellulose agar, starch
agar and solid standard medium.
Preparation of Media
One hundred grams of plane agar and 3.0 grams
of starch were dissolved in 1000ml of distilled water. The
mixture was autoclaved at 121ºc for 15 minutes and
allowed to cool for 45ºc before dispensing 20cm³ portions
into sterile petridish appropriately labeled for the test
organism. The plates were aseptically inoculated with the
test organism by streaking across the surface of the
medium; the plates were then incubated at 37ºc for 3-5
days: At the end of the incubation, the plates were
flooded with lugol’s iodine. Hydrolysis of starch was
indicated by clear zone around the colonies of the
organisms which appeared whites and the organism
blue-black for positive test while for a negative result both
the surrounding and the colony gave a blue-black
coloration, 0.5g of yeast extract, 20g of glucose, 20g of
Agar were dissolved in 1 litre of distilled water, and
heated to dissolve completely. pH maintained at 6.8,
medium was treated with actidione (cyclohexane) to
o
inhibit yeast growth before autoclaving at 121 c for 15
minutes (Obire, 2005; Schwarz, 2001).
Isolation and Identification
The samples from the cut rumen of ram, cow and
Studies on cellulose degrading microorganisms associated with rumen of ruminant animals
World J. Microbiol.
goat were label AR, BC and CG respectively Using
different sterile syringes 1ml of the liquid content from
each rumen were serially diluted by placing in test tube
containing 9ml distilled water. From the first tube, another
1ml was put to test tube 2, this was repeated through to
test tube 6.
Bacteria and fungi were isolated from the samples by
th
th
plate method, inoculating 0.1ml of the 4 and 6 dilution
factor of the serially diluted digested food from AR, BC
and CG on to nutrient agar for the isolation of bacteria,
and SDA for the isolation of fungi. The nutrient agar
0
plates were incubated at 37 c for 24hrs (aerobically and
anaerobically) while SDA plates were incubated at room
0
temperature (28 ± 2 c) for 72hrs (Oyeleke and Okusanmi,
2008) colonies that developed on the plates were used to
obtained a pure culture which was maintained on agar
slants. The pure isolates were subculture on to cellulose
agar plates and were incubated aerobically and
anaerobically for 7 days. Filter paper was use as source
of cellulose; to formulate cellulose agar; agar was
o
prepared and sterilized at 121 c for 15min, filter paper
was also sterilized in hot air oven. After cooling the plane
agar was dispensed in the petridishes and 1 filter paper
was placed in each petridish and was left until the media
had solidified (Lee and Don, 1997). The plates were
labeled appropriately and each was aseptically inoculated
with the test organism and incubated at room
temperature for 7 days, aerobically and anaerobically.
(Organisms with the ability to grow on the surface of the
filter paper were cellulose hydrolyzing bacteria fungi)
Such organisms were sub cultured again. Identification of
the bacterial isolates was based on cell morphology gram
staining, biochemical tests (motility citrate, in dole,
catalase, coagulase, oxidase, MR, VP, carbohydrate
fermentation tests, starch hydrolysis and cellulose
hydrolysis tests as described in item 3.4.1 and 3.4.2.
While the identification of fungal isolates was by
microscopic and macroscopic techniques which
comprises of pigmentation, colour of aerial shape and
kind of asexual spore presence of special structures such
as foot cell, sporangiophore or conidiophores and the
characteristics of spore. Identification was described by
Oyeleke and Manga (2008) and also Okusanmi (2008).
The identified organisms were maintained on
agar slants. A smear was prepared by placing a drop of
normal saline on a clean slide, wire loop was heated until
red hot and used to pick the growth and placed on the
slide. The smear was heat fixed by passing the slide over
flame. The smear was covered with crystal violet for one
minute and washed with distilled water. This was followed
by covering the smear with lugol’s iodine solution for one
minute. The smear was washed with distilled water and
then decolorized with acetone for 30 seconds. It was
further washed off with distilled water and the smear was
027
flooded with saffranin for 1 minute after which it was
washed with distilled water and allowed to dry. A drop of
immersion oil was placed on dried slides and the slides
were viewed under microscope using x100 objectives.
Biochemical Test
The confirmatory biochemical test described by
Oyeleke and Manga (2008) and Okusanmi (2008) were
carried out on the isolates as follows:
The motility medium contains low concentration of agar
(0.2 – 5%) and motile organisms are able to move from
the line of inoculation through the sloppy agar. The
organisms were inoculated by making a fine stab with
needle to a depth of 1-2 cm short of the bottom of the
o
tube. The organisms were incubated at 35 C for 24 – 48
hours. The line of inoculation was not sharply defined and
the rest of the medium was cloudy in positive test. In
negative result the line of inoculation became sharply
defined.
Methyl red test
Ten milliliters of MR-Vp medium in a test tube
was inoculated with the test organism and incubated at
35ºc for 48 hours, five drops of methyl red indicator
solution was added. The appearances of a red color
indicated a positive result. While yellow indicated
negative
Voges – proscauer test
Five milliliters of MR – Vp medium was
inoculated with the test organisms and incubated at 35ºc
for 48 hours. Then 0.6 ml of alpha naphthol solution and
0.2 milliliter of 40% potassium hydroxide solution were
added to the mixture. It was kept for 3 hours and
observed. The development of a red color indicated
positive result. Absence of red colour indicated negative
result.
Citrate utilization test
The isolates were inoculated in a simmon’s
citrate agar slants and incubated at 37ºc for 72 hours, the
development of a blue color indicated positive result, i.e
citrate was utilized but the presence of the original green
colour indicated a negative result i.e citrate was not
utilized.
Indole test
The isolates were inoculated in a pepton water
medium and incubated at 37ºc for 24 hours. After the
Studies on cellulose degrading microorganisms associated with rumen of ruminant animals
Sambo and Salau
028
incubation period, a drops of kovac’s reagents was
added, the appearance of a red color indicated positive
result.
Catalase test
A drop of H2O2 was placed on a sterile glass slide
and a colony of the test organisms emulsified by mixing it
with the H2O2 using a glass rod positive test was
indicated by bubbling and frothing, the absence of
bubbling indicated negative test.
Urease test
The isolates were inoculated in a urease agar
and incubated at 30ºc for 48 hours the development of
red – pink color indicated positive result.
Coagulase test
Two drops of physiological saline were placed on
a sterile glass slide and a colony of the test organism was
emulsified in it. A loop full of human plasma was added to
the mixture and mixed thoroughly. The slide was held
and titled back and forth for a minute. A positive test was
indicated by clumping of the mixture.
H2S, gas production, glucose and lactoce
The test organism was inoculated into already
prepared tubes of triple sugar iron agar and Incubated at
37ºc for 48 hours. After the incubation period the tubes
were observed for H2S production (black coloration) gas
production was indicated by (crack media, air bubbles, or
media protrude), glucose positive gave rise to a yellow
coloration at the booth of the test tube. While lactose
positive gave a redish coloration of the slant
Oxidase test
A piece of filter paper was placed in a sterile
petridish and 2 drops of freshly prepared oxidase reagent
was added. A colony of the test organisms was smeared
on the filter paper. The development of blue-purple color
within few seconds indicated a positive result. Absence of
blue purple colour indicated negative result.
RESULTS AND DISCUSSION
Eleven bacterial species i.e Bacteroides,
Streptococcus, Staphylococcus, Bacillus, Alkalegenes,
Nesseria, Peptococcus,
Lactobacillus, Veillonella,
Clostridium and Yersinia were isolated from the rumen
contents of ruminants with Bateroides spp common to all
animals, Morphological and biochemical characteristics
were used to identify the organisms see table 1. Four
fungal species namely Aspergillus flavus, Aspergillus
fumigatus, Scopulariopsis candida, and Candida
guilliermondii were also isolated from the rumen of all
animals. Macroscopic and microscopic characteristics
were used to identify the isolate as summarized in table 2
Figure 1 show the percentage occurrence of
bacterial isolates in the rumen of ram, cow and goat
respectively, and also the overall percentage occurrence
of bacterial isolates in the rumen of the ruminants.
Bacteroides and Staphylococcus spp had the highest
percentage of occurrence in ram of 33.3% each. Other
species i.e Bacillus and Veillonella had 16.6% each. In
cow Streptococcus and staphylococcus were the
dominants (22.2%), while other species i.e Bacillus,
Neserria, Peptococcus, Yesinia and Bacteroides had
uniform distribution of (11.1%) each. In goat all the
species isolated had equal distribution i.e (12.5%), and
goat rumen had the highest number of species including
Bacillus, Lactobacillus, Staphylococcus, Alcaligenes,
Yesinia, Clostridium, Veilonella and Bacteroides. On the
overall Staphylococcus had the highest percentage of
21.7% followed by Bacteroides species with 17.3% while
Streptococcus, Veilonella and Yesinia species had 8.69%
each. Alcaligenes, Yesinia and Clostridium Peptococcus
and Lactobacillus had the least percentage of 4.3% each.
Bacteroides species had the highest growth on cellulose
agar which were present in both ram, cow and goat
ruminal contents. Followed by Streptococcus found in
cow which had a moderate growth. Peptococcus spp and
Clostridium also had a moderate growth on cellulose
agar. Alcalegenes species and Nesseria had the least
growth while Bacillus and Veilonella had no growth
(Table 3).
However figure 2 indicted the percentage of fungal isolate
in the rumen of ram, cow and goat whereby in ram A.
flavus was not isolated, A. fumigatus recorded 25% and
Scopulariopsis Candida was dominants with 62.5%
Candida guilliermondii had 12.5%. In the rumen of cow
the most dominant fungus is A. fumigatus with 87.5%, A.
flavus 8.3% while S. Candida had the least percentage
4.2%. However, C. guilliermondii was not isolated. Also in
the rumen of goat A. fumigatus had the highest
percentage 46% A. flavus and C. guilliermondii had 23%
each while S. candida had 8%. In the three ruminants the
most prevalent are A. fumigatus with 64%, followed by S.
candida 16% A. flavus 11% and C. guiliermondii (9%).
For the fungal isolate, Aspergillus flavus and Aspergillus
fumigatus had the maximum cellulose degrading
capability Scopulariopsis candida had the least
Studies on cellulose degrading microorganisms associated with rumen of ruminant animals
World J. Microbiol.
029
Table 1. Summary of morphological and biochemical characteristics of bacteria isolates from the rumen of ruminants.
Sample
Gram
Reaction
MOT
MR
VP
CIT
IND
CAT
URE
COA
H2S
GAS
GLU
LAC
MAN
OX
AR3
AR2
BC1
BC4
BC1
BC2
BC3
CG5
BC2
AR1
BC4
CG1
CG3
CG4
AR4
BC3
CG3
CG2
CG6
AR2
AR3
BC5
CG4
G+cocci
G+cocci
G+cocci
G+cocci
G+cocci
G+cocci
G+coocci
G+cocci
G-cocci
G-cocci
G-shortrods
G-shortrods
G-cocci
G+rod
G+rod
G+rod
G+rod
G+rod
G-rod
G-rod
G-rod
G-rod
G-rod
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
-ve
-ve
-ve
+ve
+ve
+ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
+ve
-ve
+ve
-ve
+ve
-ve
-ve
-ve
+ve
-ve
-ve
+ve
+ve
+ve
+ve
-ve
-ve
-ve
-ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
-ve
+ve
+ve
-ve
-ve
+ve
-ve
-ve
+ve
+ve
-ve
+ve
+ve
-ve
-ve
-ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
-ve
-ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
+ve
-ve
-ve
-ve
+ve
+ve
+ve
-ve
-ve
+ve
+ve
-ve
-ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
+ve
-ve
-ve
+ve
-ve
-ve
-ve
-ve
+ve
+ve
-ve
-ve
+ve
+ve
-ve
+ve
-ve
+ve
+ve
+ve
+ve
+ve
-ve
ve
-ve
-ve
+ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
-ve
-ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
-ve
+ve
-ve
-ve
+ve
+ve
+ve
+ve
+ve
+ve
+ve
-ve
+ve
+ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
+ve
+ve
+ve
+ve
-ve
-ve
-ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
+ve
-ve
-ve
+ve
+ve
-ve
+ve
+ve
+ve
+ve
+ve
-ve
-ve
-ve
-ve
+ve
-ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
+ve
+ve
+ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
-ve
growth but Candida guilliermondii showed no
growth on cellulose agar (Table 4)
Bacteria was found to be the most
predominant, because eleven bacterial species
were isolated from the ruminants (ram, cow, and
goat) out of which nine were found to hydrolysed
cellulose (Table 4).
This agrees with findings of Bhat (2000) that,
bacteria are the most numerous of the rumen
inhabitants, there are 60 – 100 species regarded
as normal flora and although each species can
only undertake a few specific tasks, the bacteria
+ve
-ve
-ve
-ve
-ve
Starch
Hydro
-ve
+ve
-ve
+ve
+ve
+ve
-ve
-ve
-ve
+ve
+ve
-ve
-ve
+ve
+ve
+ve
-ve
-ve
+ve
-ve
+ve
+ve
+ve
Cellulose
Hydrol.
Identification
-ve
+ve
+ve
+ve
-ve
-ve
-ve
-ve
+ve
-ve
+ve
+ve
-ve
+ve
-ve
+ve
-ve
+ve
+ve
+ve
-ve
+ve
+ve
S. areus
S .arlettae
Strep-fae calis
Strep spp
Peptococcus spp
Staph inter- medius
Staph spp
Staph spp
N. haemolysans
Veillonella spp
Yersinia enterocolitica
yersinia intermedia
Vellonela spp
H. lacto bacillillus
B. lentus
Bacillus spp
B. panto- ther ticus
clostridium histolyticun
alkaligenes spp
Bacteroides convexus
Bacteroides convexus
Bacteroides succinogene
Bacteroide ovat
as a whole are capable of degrading all the
constituents of plant based diet.
Figure 1 indicated the percentage of occurrence
of each specie isolated in ram, cow and goat
respectively, in the ruminal content of ram four
bacterial specie were isolated Bacteriodes and
Staphylococcus had the highest percentage
had
Studies on cellulose degrading microorganisms associated with rumen of ruminant animals
Sambo and Salau
030
Table 2. Summary of morphological characteristic of fungi from the rumen of Ram, Cow and Goat
Sample
FGS1
Macroscopy
Light green and powder
FGS2
Bluish green and velvely
FGS3
Snow white and velvely
to powdery
FGS4
White – yellow to cream
Microscopy
branching hyphae with multinucleated mycilia
bearing string of beads of yellow green septate
conidiphores.
Aerial hyphae, bearing smooth and green
conidiophores, with head hypically columnar
Aerial hyphae with conidiogenous cells
annalete Smooth walled, whitish to creamish
conidiosphores in mass.
Pseudomycelium, with plastospores oral to
cylindrical guilliermond
Identified organism
Aspergillus flavus
Aspergillus fumigatus
Scopulariopsis candida
Candida
Table 3. Growth of bacteria on cellulose agar
Bacteria
Bacteroides spp I II and III
Streptococcus spp
Peptococcus spp
Homofermentative lactobacillus
Clostridium spp
Alcaligens spp
Nesseria spp
Yersinia spp
Iand II
Staphylococcus spp
Bacillus spp
Veillonella spp
Growth
+++
++
++
++
++
+
+
++
++
-ve
-ve
Table 4. Growth of fungi on cellulose agar
Fungi
Aspergillus flavus
Aspergillus fumigatus
Scopulariopsis candida
Candida guilliermondii
(33.3%) each while Veillonella and Bacillus had 16.6%
each. Seven bacterial species were isolated from cow
with Streptococcus having (22.2%) Staphylococcusalco
also (22.2%) while Bacteroides, Yersinia, Peptococcus,
Nesseria and Bacillus had equal distribution. Goat had
eight bacterial specie including Bacteroides, Veillonella,
Clostridium, Yersinia, Staphylococcus, Homofermentative
Lactobacillus Alcaligens and Bacillus all of which had
equal distribution. However, Bacteroides and Bacillus are
common in all rumen (ram, cow and goat) with
Bacteroides, being more prevalent in the ram. While
Streptococcus was not isolated from the ruminal content
of ram and goat it was found to be present in cow and
this is in line with the findings of Oyeleke and Okusanmi
(2008)
where
the
percentage
of
occurrence
Streptococcus overwhelm the percentage of occurrence
of all other isolates in the rumen of cow. Veillonella was
isolated from both ram and goat ruminal content but was
Growth
+++
+++
+
-
absent in that of cow. As the study revealed the presence
of Bacteroides and Streptococcus as cellulose
hydrolyzing bacteria, this agrees with Lynd, et al (2002),
and Oyeleke and Okusanmi (2008) who isolated those
organisms and implicated them in the hydrolysis of
cellulose.
The presence of Bacillus in the ruminants as noncellulose hydrolyzing bacteria except Homofermentative
lactobacillus is contrary to Lynd et al (2002), Oyeleke and
Okusanmi (2008) who reported Bacillus as hydrolyzing
organism. Apart from Staphylococcus population
encountered, Bacteroides were the most highly prevalent
in the ruminants with (17.3%) followed by Bacillus (13.0)
and then Streptococcus and Yersinia (8.69%) each in
(table 4) Fungi also inhabit the rumen and are known to
play an important role in the degradation of
polysaccharides other than cellulose as strengthening
Studies on cellulose degrading microorganisms associated with rumen of ruminant animals
World J. Microbiol.
031
Figure 1. Occurrence of Bacterial isolates in the Rumen of Ruminant
Figure 2. Occurrence of Fungal Isolates in the rumen of Ruminant
agents in the cell wall of woody plants (Madigan et al,
2000) In this research four fungal species were isolated
from the rumen of the ruminants and their presence and
percentages of occurrence were represented in figure
4.2, i.e for ram, cow and goat and also the overall
percentages of occurrence. A. fumigatus was observed to
be common in all the rumens, but had the highest
percentage of occurrence 87.5% in cow, represented the
highest percentage of (64%) on the overall. A. flavus was
not found in ram but was isolated from cow, and goat.
However, S. scopulariopsis, appeared in all rumen, but
more prevalence in ram (62.5%), C. guilliermondii
appeared in ram and goat but more in goat. Based on
cellulose degradation capability (Table 4) A. fumigatus
and A. flavus had the maximum cellulose degradation.
However the fungal species isolated is related to the
previous findings of Masoni et al (1997) where he
reported that members of the division Deutromyces e.g
those of the genera Aspergillus have cellulolytic
enzymes; Oyeleke and Okusanmi (2008) isolated A.
flavus from rumen of ruminants and implicated it in
cellulose degradation.
CONCLUSION
The study revealed that ruminants (ram, cow, and goat)
harbors various organisms that are active cellulose
Studies on cellulose degrading microorganisms associated with rumen of ruminant animals
Sambo and Salau
032
degraders, out of which Bacteroides specie grow best on
cellulose agar. Rumen should be a site for isolation of
micro organisms that are capable of cellulose hydrolysis
in order to reduce the coast of purchasing commercial
enzymes.
ACKNOWLEDGEMENT
The authors acknowledged the contributions of Dr. Anshu
S Chatli, Saud Hamidovic, Ewa Karwowska, Prof. Dr.
Mohamed S. Abdel-Aziz, Dr. Jonatas Rafael de Oliveira,
Mostafa Essam Eissa and Dimitra Dimitriadi for donating
their time, critical evaluation, constructive comments, and
invaluable assistance toward the improvement of this
very manuscript.
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Accepted 24 July, 2015.
Citation: Sambo S, Salau IA (2015). Studies on cellulose
degrading microorganisms associated with rumen of the
ruminant animal. World Journal of Microbiology, 2(2):
026-032.
Copyright: © 2015 Sambo and Salau. This is an openaccess article distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium,
provided the original author and source are cited.
Studies on cellulose degrading microorganisms associated with rumen of ruminant animals