ISSN: 19067976

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[[[

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16S rRNA
. 1

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36

.. 43

. 46

63

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16S rRNA
(The use of 16S rRNA probes Technique for identifying cellulolytic bacteria in the rumen)

Butyrivibrio, Fibrobacter Ruminococcus spp. Fibrobacter Ruminococcus spp.
Butyrivibrio fibrisolvens



16S rRNA probes
5 3 16S rRNA
probes Butyrivibrio fibrisolvens, Ruminococcus albus strains R. flavefaciens
strains inoculated Ruminococcus

Fibrobacter Ruminococcus spp.

: 16

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Abstract
Based on reviewed literature, it was found that fiber (lignocellulose) digestion is an anaerobic process
in ruminants and usually takes place in the rumen. Several genera of bacteria are involved in fiber degradation
and it is primarily carried out by Butyrivibrio, Fibrobacter and Ruminococcus spp. Fibrobacter and
Ruminococcus spp. are the most fibrolytic, possibly related to their adherent ability, while Butyrivibrio
fibrisolvens is a free-floating organism. The 16S rRNA oligonucleotide probe was specific of sequence from
5 to 3 for the species or genus level. 16S rRNA analysis has provided a framework for evaluating microbial
communities and has provided significant insights into a variety. According to this technique, inoculation of
Ruminococcus into rumen of lamb, it was found there were increases of Ruminococcus, Fibrobacter
succinogenes, rumen fungi and decrease of gram-positive bacteria in the rumen of lambs. In addition evidence
of how the microbial community is affected by manipulation, especially with respect to the fibrolytic
populations and should improve ruminants production in the future.
Key words: 16S rRNA probes, Butyrivibrio spp., Fibrobacter spp., Ruminococcus spp.

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Butyrivibrio,
Fibrobacter Ruminococcus spp. Fibrobacter Ruminococcus spp.

Butyrivibrio
fibrisolvens (rumen fluid)
(Weimer, 1996)
rumen pH 6.5-7 39
17-20 mg% (Wanapat, 2000)

Sijpelsteijn (1951) Ruminococcus flavafaciens
cellulose cocci
Hungate (1957) R. albus
diplococci

(volatile fatty acid, VFAs)


phenotypic phylogenetic Fibrobacter 16S rRNA
(Lin and Stahl, 1995; Forster et al., 1997)
16S rRNA probes
1
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(Hungate, 1966; Leedle et a., 1982) 16S rRNA probes

(Amann et al., 1995)

probes (Pace, 1997) Fibrobacter
Ruminococcus spp. 16S rRNA probes
oligonucleotide probes Ruminococci
(Odenyo et al.,
1994a,b)
16S rRNA probes

16S rRNA
rRNA (ribosomal ribonucleic acid) RNA
65 80 RNA
rRNA 4 28S, 18S, 5.8S 5S
3 23S, 16S 5S ( 1)
DNA rRNA
rRNA DNA
rRNA
(Champe and Harvey, 1994)
16S rRNA probes
16S rRNA probes
Odenyo
et al. (1994 a,b) Krause et al. (1999, 2000)

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1.
(basal medium)

39 C
2. 16S rRNA
16S rRNA probes

3. 16S rRNA probes

PCR (polymerase chain reaction)

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A. Prokaryotic
ribosome

B. Eukaryotic
ribosome
1
A = 70S (Prokaryotic ribosome)
B = 80S (Eukaryotic ribosome)
: Lehninger et al. (1993)

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16S rRNA probes (cellulolytic bacteria)

Alm et al. (1996) Krause et al. (2000) 16S rRNA probes

5 3 ( 1)
Froster et al. (1997) Krause et al. (1999) 16S rRNA probes
Butyrivibrio fibrisolvens strains, Ruminococcus albus strains R. flavefaciens strains

( 2)

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1 oligonucleotide probe
Oligonucleotide probe
name
S-G-Fibr-0153-a-S-16
S-Ss-F.s.suc-0628-a-A-22

Probe sequence from

Target group
5 to 3
CCGTGCCAACGCGCGG Genus Fibrobacter

References
Alm et al.
(1996)

GATCCAGTTCGGACTG Subspecies Fibrobacter

CAGAGC
succinogenes subsp.
Succinogenes
S-Ss-F.suc(2)-0628-a-A-22 AACCCAGTTCGGACTG Subspecies F.
CAGGTC
succinogenes group 2
S-Ss-Ralb SY3-0174-a-S-20 GATAAGCCGCATGACC R. albus SY3
Krause et al.
TATC
(2000)
S-F-Fungi-0715-a-A-25
GTACACACAATGAAGT Anaerobic rumen fungi
GCATAAAGG
S-S-Fsucc-0649-a-A-20
TGCCCCTGAACTATCC
F. succinogenes
AAGA
S-G-Rum-IV-0132-a-A-17 RTTATGRGGTATTAMC R. albus + R. flavefaciens
in cluster IV of clostridia
A

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2 Ruminococcus strains
Strain
Butyrivibrio fibrisolvens1
49
H10b
H17c
UC12254
Ruminococcus albus strains2
B199
AR67
Ra8
SY 3
R. flavefaciens strains2
FD-1
LP-9155
R13e2
RF1Ba
AR 72
AR 71
Y1
AR69
AR46
AR47
B146
R1-addax
C14-addax
: 1 = Froster et al. (1997)
2 = Krause et al. (1999)

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Source/host
Bovine
Bovine
Bovine
Bovine
Cow
Sheep
Cow
Sheep
Tablet containing ruminal organisms
Cattle
Reindeer
Sheep
Sheep
Sheep
Sheep
Sheep
Sheep
Sheep
Cow
Addax
Addax


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16S rRNA probes (cellulolytic bacteria)

Froster et al. (1997) cellulolytic bacteria
probes 49, 156, 189, D1 D1-b
Ruminococcus albus SY3, R. flavefaciens
Fibrobacter succinogenes S85 Krause et al. (1999)

R. flavefaciens, R. albus , B.
fibrisolvens F. succinongenes
( 3)
3 cellulolytic bacteria
Item

Mean
SD

Microscopic and culturable counts

(x109/g digesta)
TCC
CEL
DC
16.3
9.6
0.5
3.9
1.4
0.2

Proportion CEL
(%)
CEL/DC
CEL/TCC
3.1
1.5

5.2
2.6

16S rRNA oligonucleotide probes (%)

F. succinogenes
Ruminococcus IV
Ruminococcus IV+F. succinogenes
Mean
2.2
1.8
4.0
SD
0.7
0.7
1.0
DC = direct counts, TCC = total culturable counts, CEL = total cellulolytic bacteria
: Krause et al. (1999)
Froster et al. (1997) Krause et al. (1999)
Krause et al. (2000) cellulolytic
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bacteria inoculated
3 1 inoculated
Ruminococcus 2 inoculated Ruminococcus
4 3 inoculated Ruminococcus
inoculated Ruminococcus
3.4 x 108-0.8 c 109 cells/ml inoculated 2
23 63 inoculated
R. albus SY3 R. albus F. succinogenes
( 4 2)
( 3)
( 4)
Wora-anu et al. (2007)

3 Fobrobacter succinogenes, Ruminococcus albus
R. flavefaciens

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4 Ruminococcus albus SY3

Daya Lamb number:
Dosed
Control
Remained with ewe
Received milk replacer
A1 A2 A3 A4 A1 A2 A3 A4 A5 A1 A2 A3 A4
63 105 109 108 109 108 107 108 107 109 107
70 107 108 107 107 108 107 106 107 109
77 105 105 107 106 105 105 105 106 107 107 106
91 105 105 106 105 105 105 105
113 105
105 135 a
Age of the lamb. Lamb were dosed from day 23 to 63 of age.
Approximate numbers of R. albus SY3 in the rumen in individual lambs (10x cells/ml)
: Krause et al. (2000)

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A5
109
106
107
105

A6
107
106
106
-


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2
A = Ruminococcus B = Fibrobacter succinogenes
: Krause et al.. (2000)

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3 Eukayotes (A) Fingi (B)

: Krause et al. (2000)

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4
: Krause et al. (2000)

phylogenetic 16S rRNA
Butyrivibrio, Fibrobacter Ruminococcus spp.





16S
rRNA probes

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Alm, E.W., D.B. Oerther, N. Larsen, D.A. Stahl and L. Raskin. 1996. The Oligonucleotide Probe database.
Appl.Environ. Microbiol. 62:3557-3559.
Amann, R.I., W. Ludwig, K.H. Schleifer. 1995. Phylogenetic identification and in situ detection of individual
microbial cells without cultivation. Microbiol. Rev. 59:143-169.
Champe, P.C. and R.A. Harvey. 1994. Lippincotts IIIustrated Reviews:Biochemistry 2nd edition. J.B.
Lippincott company, Philadelphia.
Forster, R.J., J. Gong and R.M. Teather. 1997. Group-specific 16S rRNA hybridization probes for
determinative and community structure studies of Butyrivibrio fibrisolvens in the rumen. Appl.
Environ. Microbiol. 63:1256-1260.
Hungate, R.E. 1957. The microorganisms in the rumen of cattle fed a constant ration. Can. J. Microbiol.
3:289-311.
Hungate, R.E. 1966. The rumen and its microbes, (Eds. By R.E. Hungate). New York : Academic Press. pp.
1-533.
Krause, D.O., B.P. Dalrymple, W.J.M. Smith, R.I. Mackie and C.S. McSweeney. 1999. 16S rRNA
sequencing Ruminococcus albus and Ruminococcus flavefaciens:design of a signature probe and its
application in adult sheep. Microbiology. 145:1797-1807.
Krause, D.O., W.J.M. Smith, F.M.E. Ryan, R.I. Mackie and C.S. McSweeney. 2000. Use of 16S-rRNA
based techniques to investigate the ecological succession of microbial populations in the immature
lamb rumen:traccking of a specific strain of inoculated Ruminoncoccus and interactions with other
microbial populations in vivo. Microb Ecol. 38:365-376.
Leedle, J.A.Z., M.P. Bryant and R.B. Hespell. 1982. Diurnal variations in bacterial numbers and fluid
parameters in ruminal contents of animals fed low-or high-forage diets. Appl. Environ. Microbiol.
44:402-412.
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Lehninger, A.L. D.L. Nelson and M.M. Cox. 1993. Principles of Biochemistry, 2nd ed. Irving Place, New
York:Worth Publishers. 324-357.
Lin, C. and D.A. Stahl. 1995. Taxon-specific probes for cellulolytic Fibrobacter reveal abundant and novel
ezuine associated populations. Appl. Environ Microbiol. 61:1348-1351.
Odenyo, A. A., R.I. Mackie, D.A. Stahl and B.A. White. 1994a. The use of 16S rRNA-targeted
oligonucleotide probes to study competition between ruminal fibrolytic bacteria:development of
probes for Ruminococcus species and evidence for bacteriocin production. Appl. Environ. Microbiol.
60:3688-3696.
Odenyo, A. A., R.I. Mackie, D.A. Stahl and B.A. White. 1994b. The use of 16S rRNA-targeted
oligonucleotide probes to study competition between ruminal fibrolytic bacteria:pure-culture studies
with cellulose and alkaline peroxide-treated wheat straw. Appl. Environ Microbiol. 60:3697-3703.
Sijpelsteijn, A.K. 1951. On Ruminococcus flavefaciens a cellulose decomposing bacterium from the rumen
of sheep and cattle. J. Gen. Microbiol. 5:869-879.
Wanapat, M. 2000. Rumen manipulation to increase the efficient use of local feed resources and
productivity of ruminants in the Tropics. In:Proc. of The 9th Congress of the Asian-Australasian
Association of Animal Production Societies and 23rd Biennial Conference of the Australian Society of
Animal Production, July 3-7, 2000. University of New South Wales, Sydney, Australia.
Weimer, P.J. 1996. Why dont ruminal bacteria digest cellulose faster. J. Dairy Sci. 79:1496-1502.
Wora-anu, S., M. Wanapat, C. Wachirapakorn and N. Nontaso. 2007. Effect of Roughage Sources
on Cellulolytic Bacteria and Rumen Ecology of Beef Cattle. Asian-Aust. J. Anim. Sci. 20(11) :
1705.

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(Role of rumen fungi on fiber digestion)

Neocallimastix sp., Piromyces sp.,
Ceacomyces sp., Orpinomyces sp. Anaeromyces sp.
endo-glucanase (CMCase) exoglucanase (cellobiohydrolase) cystalline
xylannase (hemicellulase)
6
sclerenchyma vascular 20-24
mesophyll, parenchyma pholem

: , , ,

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Abstract

Based on reviewed literature it was found that anaerobic fungi are now well recognized as one of
the major components of rumen microorganisms. It is certain that they have roles in ruminal fiber digestion.
Five genera have been established so far namely; Neocallimastix, Piromyces, Caecomyces, Orpinomyces and
Anaeromyces. Rumen fungi produce cellulase that consists of a complex of enzymes capable of sulubilising
both the amorphous and cystalline cellulose present in plant tissue. Large amount of endo-glucanase
(CMCase) and exo-glucanase (cellubiohydrolases), which can degrade crystalline cellulose is also produced
but in lesser amount the endoglucanase; however it was found that rumen fungi produce xylanase
(hemicellulase) also. In colonising plant tissues anaerobic rumen fungi showed that by 6 hr lignified tissue
such as sclerenchyma and vascular tissue are extensively colonized. After attachment and encystment the
zoospore geminates with a single germ-tube which penetrates the plant tissue. The germ-tube elongates and
branches very rapidly forming a network of rhizoid after 20-24 hr of incubation. Most of the colonised thinwalled tissue such as the mesophyll, parenchyma and phloem are degraded. The end-products were able to
ferment cellulose producing formate, acetate, lactate, ethanol, carbondioxide and hydrogen. However, the
fermentation end-products are dependable on rumen fungi population and type of feeds in order to increase
ruminant production efficiency.
Key words: rumen fungi, fiber digestion, enzymes, fermentation end-products.

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- (pH)
6.5-7.0
effective fiber
(volatile fatty acid, VFA) (acetic acid, C2)
(propionic acid, C3) (butyric acid, C4)

(non-protein nitrogen, NPN)
-
(Wanapat, 2000)


(polyflagellate) Orpin (1976) Ho and Barr (1995)
Piromonas communis

103-104 /.
(Rumen fluid) (Jouany and Ushida, 1999)
endo-1,4--D-glucanase (CMCase)
(Mountfort and Asher, 1985), exo-1-4--D-glucanase (cellobiohydrolases)
cystalline hemicellulose
xylanases (hemicellulase) -xylanase, -xylosidase xylobiase (William and
Orpin, 1987) Xue et al. (1992a, b) cellulase
xylanase
Neocallimastix Orpinomyces
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endo-pectinylase Wanapat et al. (2000)

Anaeromyces
sp. 7.30 x 106 cells/ml



6

thallus
thallus 2 monocentric polycentric monocentric
polycentric
5 (Ho and Barr, 1995)
1. Neocallimastix tallus monocentric
(elongate-ovoid) 2-25 (polyflagellate)
2. Piromyces tallus monocentric 2-4
(uniflagellate)
3. Caecomyces tallus monocentric polycentric
2-4
4. Anaeromyces tallus polycentric
(acuminate apex)
5. Orpinomyces 4-25 (polyflagellate) tallus
polycentric

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Ushida et al. (1997)

Ho and Barr (1995) Table 1
Table 1. Classification of anaerobic fungi in the rumen.
Genera and characteristics
Neocallimastic
Monocentric, polyflagellate zoospore,
extensive, Filamentous rhizomycelium

Piromyces
Monocentric, monoflagellate zoospore,
Filamentous rhizomycelium

Opinomyces
Polycentric, polyflagellate zoospore
Filamentous rhizomycelium
Caecomyces
Polycentric or monocentric,
monoflagellate zoospore ,
spherical hold fasts
Anaeromyces
Polycentric, monoflagellate zoospore,
Filamentous rhizomycelium

Source: Ho and Barr (1995); Ushida et al. (1997)

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Species

Source

Frontalis
Patriciarum
Hurleyensis
Variabilis

sheep
sheep
sheep
sheep

Communis
Mae
Dumbonicus
Rizinflatus
Minutus
Spiralis
Citronii

sheep
horse
elephant
ass
deer
goat
horse

Joyonii
Intercalaris

sheep
cattle

Communis
Equi

sheep
horse

Mucronatus
Elegans

cow
sheep


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(receptor) glucose, sucrose, mannose
(Orpin and Boutiff, 1978) Wubah and Kim (1996)
Neocallimastic frontalis, Piromyces communis, Orpinomyces joyonii Anaeromyces sp.
phenolic acid -coumaric acid, ferulic acid syringic acid
(lignin)
Matsui et al. (1997)

stomata (Bauchop, 1981; Akin
et al., 1983; Ho et al., 1988a,b, 1991)
15-30 ( 1A) (Ho et al., 1988b) single
germ tube ( 1B)
(Ho et al., 1996)

(cellulase) (cellulolse)

(Ho et al., 1991) Wanapat et
al. (2000)

- (digesta)

( 2)
7.30 x 106 3.78 x 106 cells/ml

6 sclerenchyma
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vascular tissue (Ho et al., 1988b) Akin et al. (1983)

Digitaria pentzii 6
mesophyll, parenchyma
phloem 24 ( 3)
extra cellular enzyme (Akin et al., 1990) Akin (1994)
(Table 2)
Akin et al. (1990)
(Table 3)

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Table 2. Dry weight loss of intact forages by rumen fungi.

Plant

Warm-season grasses
Cynodon dactylon (Bermuda grass)
Sorghum bicolor (sorghum)

Cool-season grasses
Phalaris arundinacea (reed canary-grass)

Festuca arundinacea (tall fescue)

Incubated with mixed rumen fungi1/

Plant part
% Dry Weight loss2/
Blade
37.6
Sheath
Stem
Blade
Sheath
Stem

38.5
31.4
36.5
36.3
25.7

Blade
Sheath
Stem
Blade

28.6
40.0
45.2
44.5
48.2

Sheath
1/
incubated 72 hours with rumen fluid plus streptomycin and penicillin.
2/
values are corrected for acid-pepsin-soluble materials and for loss in unenoculated controls.
Source : Akin (1994)

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1 A =
stoma 30 (Bar = 5
m) (Ho et al., 1988b)
B = parenchyma
6 (Bar = 2 m)
(Ho et al., 1996)

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2
A =
Anaeromyces sp., B =
(Bar = 16 m)
Source : Wanapat et al. (2000)

A B
3 A =
24
(Bar = 25 m) (Ho et al., 1988b)
B = 24
(Bar = 25 m) (Ho et al., 1996)
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Table 3. Effect of rumen fungi on dry weight loss of Bermuda grass.

Fungi

Bermuda grass incubated with pure rumen fungal culture1/

Plant part
% Dry Weight loss

Monocentric
Piromyces MC-1
Neocallimastix MC-2

Polycentric
PC-1 (probably Anaeromyces)
Orpinomyces PC-2
Orpinomyces PC-3

Blade
Stem
Blade
Stem

29.5
15.7
67.8
20.6

Blade
Stem
Blade
Stem
Blade
Stem

42.5
16.2
56.0
23.8
57.2
33.6

1/

incubated 72 hours in basal medium.

Source : Akin et al. (1990)

Ha et al. (2001)

Piromyces rhizinflata B157 (dry matter digestibility, DMD)
50.54% Ruminococcus flavefaciens FD1
47.78 % P. rhizinflata + R.
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flavefaciens Orpinomyces joyonii + R. flavefaciens 44.05

41.73 % 6 (Table 4)
Table 4. Rice straw degradation by rumen fungi and rumen cellulolytic bacteria in mono- or in coculture (%)
Culture

Incubation time (days)

2
4
26.99+2.48
22.70+0.66d
27.68+1.00
41.23+0.41a
26.93+1.90
24.70+0.86cd
26.08+0.79
28.58+1.76c
24.49+0.29
45.96+1.66a
24.79+0.43
29.09+0.74bc
25.15+0.28
26.41+1.35cd
24.57+0.92
33.82+0.94b
24.53+0.97
28.25+0.74c
27.77+0.80
27.34+0.66cd
24.62+0.80
44.35+2.53a

1
Orpinomyces joyonii SG4
26.50+0.95
Piromyces rhizinflata B157
24.75+0.60
Fibrobacter succinogenes S85
21.47+1.51
Ruminococcus albus B199
21.01+0.78
Ruminococcus flavefaciens FD1 22.95+0.39
P. rhizinflata + F. succinogenes 21.72+0.25
P. rhizinflata + R. albus
22.85+0.73
P. rhizinflata + R. flavefaciens
22.71+1.07
O. joyonii + F. succinogenes
22.49+1.13
O. joyonii + R. albus
23.47+0.76
O. joyonii + R. flavefaciens
26.47+4.16
Dry matter digestibility 0 hr is 15.67+0.54%,
Means in the same column with different superscripts are significant different (p<0.01)
Source : Ha et al. (2001)

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6
31.19+0.60cd
50.54+3.44a
33.56+0.51bc
30.03+1.03d
47.78+0.65a
31.65+1.55cd
29.54+1.14d
44.05+2.32ab
31.76+0.83cd
31.33+1.36cd
41.73+5.83abc


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N. frontalis endo-1,4--D-glucanase (CMCase) (Mountfort and Asher, 1985)

N. patriciarum, P. communis
(William and Orpin, 1987) P. spirals, P. minutus (Ho et al., 1996) Anaeromyces elehans (Ho et al.,
1990) exo-1-4--D-glucanase (cellobiohydrolases)
cystalline endoglucanase (Mountford et al.,
1982)
hemicellulose xylanases
(hemicellulase) -xylanase, -xylosidase xylobiase
soluble sugar (William and Orpin, 1987) Xue et al. (1992a,b)
cellulase xylanase
Ushida et al. (1997)
polysaccharidases
Table 5

acetate, formate,
lactate, ethanol, CO2 H2 Neocallimastix frontalis, N. hurleyensis, Caecomyces
sp., Piromyces sp. (Bauchop and Mountfort, 1981) N. patriciarum
acetate, lactate, H2 CO2 Piromyces P. spiralis P.
minutus acetate formate (Ho and Abdullah, 1999)
acetate
lactate propionate
(, 2533)

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Table 5. List of sequence data on fungal polysaccharidases.

Organism
Neocallimastix patriciarum
N. patriciarum
N. frontalis
N. frontalis
N. frontalis
N. patriciarum

Enzyme
Endoglucanase
Xylanase
Endoglucanase
Endoglucanase
Endoglucanase

N. patriciarum
Piromyces sp.
Piromyces sp.
Piromyces sp.
Piromyces sp.
Orpinomyces sp.
Orpinomyces sp.
Orpinomyces
Orpinomyces
Source : Ushida et al. (1997)

Cellobiohydrolase
Mannanase
Endomannanase
Endomannanase
Xylanase
Xylanase
Endoglucanase
Cellobiohydrolase
Cellobiohydrolase

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Xylanase

Gene
celB
xynA
celA
xynl
xyn2
xynB
celA
manA
manB
manC
xynA
xynA
celB
celA
celC


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Neocallimastix sp., Piromyces sp. Orpinomyces sp. endo-glucanase
(CMCase) Neocallimastix sp. Orpinomyces sp.
exo-glucanase cystalline
hemicellulase

(H-sink)

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. 2533. . : .
Akin, D.E. 1994. Ultrastructure of plant cell-walls degraded by an aerobic fungi:In Anaerobic Fungi.
Biology, Ecology, Function. Eds. D.O. Mountfort and C.G. Orpin. Macel Dekker, New York.
Akin, D.E., G.I.R. Gordon, and J.P. Hogan. 1983. Rumen bacterial and fungal degradation of Digiaria
pentzii grow with or without sulphur. Appl. Environ. Microbiol. 46:738-748.
Akin, D.E., L.L. Rigsby, C.E. Lyon, and W.R. Widham. 1990. Relationship of tissue digestion to textural
strength in bermuda grass and alfulfa stems. Crop. Sci. 30:990-993.
Bauchop, T. 1981. The anaerobic fungi in rumen fibre digestion. Agric. Environ. 6:339-348.
Bauchop, T., and D.O. Mountfort. 1981. Cellulose fermentation by a rumen anaerobic fungus in both the
absence and presence of rumen methanobens. Appl. Environ. Microbiol. 42:1103-1110.
Ha, J.K., S.S. Lee, S. W. Kim, In K. Han, K. Ushida, and K.J. Cheng. 2001. Degradation of rice straw by
rumen fungi and cellulolytic bacteria through mono-, co- or sequential- cultures. Asian-Aus. J. Anim.
Sci. 14:797-802.
Ho, Y.W., and N. Abdullah. 1999. The role of rumen fungi in fibre digestion, review. Asian-Aus. J.
Anim. Sci. 12:104-112.
Ho., Y.W., and D.J.S. Barr. 1995. Classification of anaerobic gut fungi from herbivores with
emphasis on rumen fungi from Malaysia. Mycologia. 87:655-677.
Ho, Y.W., N. Abdullah, and S. Jalaludin. 1988a. Penetrating structures of anaerobic rumen fungi in
cattle and swamp buffaloes. J. Gen. Microbiol. 134:177-181.
Ho, Y.W., N. Abdullah, and S. Jalaludin. 1988b. Colonization of guinea grass by anaerobic rumen
fungi in swamp buffaloes and cattle. Anim. Feed Sci. Technol. 22:161-171.
Ho, Y.W., N. Abdullah, and S. Jalaludin. 1991. Fungal colonization of rice straw and palm press fiber
in the rumen of cattle and buffaloes. Anim. Feed Sci. Technol. 34:311-321.
Ho, Y.W., N. Abdullah, and S. Jalaludin. 1996. Microbial colonisation and degradation of some
fibrous crop residues in the rumen of goats. Asian-Aus. J. Anim. Sci. 9:519-524.
Ho, Y.W., T. Bauchop, N. Abdullah, and S. Jalaludin. 1990. Ruminomyces elegans gen. et sp. nov. ,
33 | P a g e


1 1 ..2553

a polycentric fungus from cattle. Mycotoxon. 38:397-405.

Jouany, J.P., and K. Ushida. 1999. The role of protozoa in feed digestion review. Asian-Aus. J.
Anim. Sci. 12:113-128.
Matsui, H., K. Ushida, and Y. Kujima. 1997. Effect of dietary concentrate on fungal zoosporogellesis
in sheep rumen. Asian-Aus. J. Anim. Sci. 10:599-602.
Mountfort, D.O., and R.A. Asher. 1985. Production and regulation of cellulase by two strains of the
rumen of the rumen anaerobic fungus Neocallimastix frontalis. Appl. Environ. Microbiol. 49:13141322.
Mountfort, D.O., R.A. Asher, and T. Bauchop. 1982. Fermentation of cellulose to methan and carbon
dioxide by a rumen anaerobic fungus in a triculture with Methanobrevibacter sp. Strain RA1 and
Methanosarcina barderi. Appl. Environ. Microbiol. 44:128-134.
Orpin, C.G., and L. Bountiff. 1978. Zoospore chemotaxis in the rumen phycomycete Neocallimastix
frontalis. J. Gen. Microbiol. 104:113-122.
Ushida, K., H. Matsui, Y. Fujino, and J.K. Ha. 1997. Role and potential of ruminal in fiber digestion.
Asian-Aus. J. Anim. Sci. 10:541-550.
Wanapat, M. 2000. Rumen manipulation to increase the efficient use of local feed resources and
productivity of ruminants in the Tropics. In:Proc. of The 9th Congress of the Asian-Australasian
Association of Animal Production Societies and 23rd Biennial Conference of the Australian Society of
Animal Production, July 3-7, 2000. University of New South Wales, Sydney, Australia.
Wanapat, M., A. Ngarmsang, S. Korkhuntot, N. Nontaso, C. Wachirapakorn, G. Beakes, and P.
Rowlinson. 2000. A comparative study on the rumen microbial population of cattle an swamp
buffalo raised under traditional village conditions in the northeast of Thailand. Asian-Aus. J. Anim.
Sci. 13:918-921.
Williams, A.G., and C.G. Orpin. 1987. Polysaccharide-degrading enzymes formed by three spicies of
anaerobic rumen fungi grown on a range of carbohydrate substrates. Can. J. Microbiol. 33:418-426.
Wubah, D.A., and S.H. Kim. 1996. Chemoattraction of anaerobic ruminal fungi zoospores to selected
phenolic acids. Microbiol. Res. 151:257-262.
34 | P a g e


1 1 ..2553

Xue, G.P., K.S. Gobius, and C.G. Orpin. 1992a. A novel polysaccharide hydrolase cDNA (cel D)
from Neocallimastix partricicum encoding three-multi-functional catalytic domains with high
endoglucanase, cellobio-hydrolase and xylanase activities. J. Gen. Microbiol. 138:2397-2403.
Xue, G.P., C.G. Orpin, K.S. Gobius, J.H. Aylward, and G.D. Simpson. 1992b. Cloning and expression of
multiple cellulase cDNAs from the anaerobic rumen fungi Neocallimastix partriciarum in Esherichia
cili. J. Gen. Microbiol. 138:1413-1420.

35 | P a g e


1 1 ..2553

(Heterosis effects on reproductive traits between Landrace and Largewhite)

7
110

36 | P a g e


1 1 ..2553

Abstract
The objective of this study was to evaluate the crossbred between Landrace and Largewhite that
which breeds should be the effective sire or dam. The seven traits of reproductive traits of dam were observed.
The results showed that age at puberty, weight at puberty, gestation length, weight at 110 days, weight at
weaning and lactation weight loss of crossbred between Landrace (dam) and Largewhite (sire) were better that
crossbred between Landrace (sire) and Largewhite (dam) and better than purebred Landrace or purebred
Largewhite. While the number of nipple and gestation lengths were similar among any types of breeds or
crossbreds.

Key words: Heterosis, Landrace, Largewhite, Crossbred and Reproductive traits

37 | P a g e


1 1 ..2553

(, 2543)


(, 2548)

, ,
Cassady et al.
(2009) 7 , ,
, , 110 ,
( 1)
,
188.402.30 , 92.601.10 ,
38 | P a g e


1 1 ..2553

136.801.70 -18.201.40 210.702.30

, 106.101.10 141.401.80 -11.001.50
110 152.301.80
155.001.70


(182.009.40 ) (91.704.40 ) 110
(159.206.20 ) (129.106.50 )
(-28.005.30 )

( 1)

39 | P a g e


1 1 ..2553

1 (SE)

14.160.06

14.290.24

13.940.22

()

14.350.06

()

188.402.30 210.702.30 187.9011.00 182.009.40

92.601.10

106.101.10 102.705.10

91.704.40

()
()

114.480.17 114.290.19 114.430.79

114.360.64

155.001.70 152.301.80 164.67.80

159.206.20

136.801.70 141.401.80 157.107.80

129.106.50

-18.201.40 -11.001.50 -7.006.30

-28.005.30

110 ()

()

()
: Cassady et al. (2002b)
40 | P a g e


1 1 ..2553

110

. 2543.. 7. . .5-3
, , . 2545.
. .. 2546.
. .111-114
, Crump .R. , Suarez .M. and Graser .H-U. 2549.
.
..
41 | P a g e


1 1 ..2553

. 2545. 3 15-16
Cassady, JP, LD Young, and K.A. Leymaster. 2002a. Heterosis and recombination effects on pig growth and
carcass traits. J.Anim. Sci. 80:22782294.
Cassady .JP, LD Young and KA Leymaster. 2002b. Heterosis and recombination effects on pig reproductive
traits. J. Anim. Sci. 80, 2303-15.
Johnson, R. K. 1980. Heterosis and breed effects in swine. NorthCentral Regional Publication .No. 206.
Nebraska AgriculturalExperiment Station, Lincoln
Ross, T.M. 1992. Canadian swine improvement program. Adsgo Trading corporation limited, 916 Grosvenor
Avenue, Winnepeg, Manitoba, Canada. 17p.

42 | P a g e


1 1 ..2553

5,000

18
56
..2331

63
..2474

43 | P a g e


1 1 ..2553

4
1. 2
2.
3.
4. 4

44 | P a g e


1 1 ..2553

, -
, -

45 | P a g e


1 1 ..2553

Pattern of learning management participatory of community philosophers

5
1 () 23 ..2550
22 ..2551 12
4
1


2

46 | P a g e


1 1 ..2553

4 -
-

1-4

: , ,

47 | P a g e


1 1 ..2553

Abstract
This research was the qualitative research; the purpose of this study was to seek for the pattern of
learning management participatory of community philosophers which administered to five leading secondary
schools in Nong Khai Province which consisted of Khokkhon Wittayakom School, Thon Wittayakom School,
Thabo School, Thabo Pittayakom School and Municipality School 1 (Sawang Wittaya). The period in this
study was from the 23rd July 2007 until the 22nd July 2008 at the total of 12 months. The finding: Pattern of
learning management participatory of community philosophers has divided into 4 steps namely:
Step 1 The beginning step; the administrator has committed the policy of learning management
participatory of community philosophers to the teachers at school. They will look for the community
philosophers, selected and planned for learning. After that the administrator and the teachers have together
presented their leaning planned towards the committees of school to asking for the procedure allowable.
Step 2 The performance step; the teacher searched for the students. The community philosophers and
the teachers have together administered the learning. The teacher will be in theory session and the community
philosophers will be in practical session. The learners participated in both theory and practical session. The
local administration organization has supported this learning management in some matters such as the
budgets, the materials etc.
Step 3 The evaluation learning step; the teachers and the community philosophers have evaluation the
learning management continuously with surveying the pleasure of the learners and the guardian towards the
learning for developed it appropriately.
Step 4 The expansion step; the teachers concluded the learning management participatory of
community philosophers to the administrator in writing, in order to make decision to expand the learning in
the future.
The learning management of school will be developed in circle. The procedure will go round back
and forth from the 1st step to the 4th step. The procession of learning management will developed own-selves
continuously and the school will interpreted them continuously.
The suggestions; it should be operated and developed it continuously until access the local
curriculum, divided the level of learning into the interval from easily to difficulty. There should be the
48 | P a g e


1 1 ..2553

research in multi-discipline style in learning management participatory of community philosophers for

integrating it completely. It should be deep studied the educational management participatory of community
philosophers in action style in order to bring the community philosophers participatory in Thai curriculum.
Keyword: Learning management, Participatory, Community Philosophers

49 | P a g e


1 1 ..2553

(Knowledge-Based Economy: KBE)

(Knowledge
Worker)
(, 2549)



(, 2542)

2542
12 3
( 15) ( 23)
( 25)



..2542
(Integration)

(Context)




50 | P a g e


1 1 ..2553

(Brain-based Learning) (School-based
Learning Education)


(Sustainable Development)

(Spirituality)
4
(World Health Organization: WHO) 4

(Holistic) 4
(, 2549)
(Social Capital)
(Community Context)
(Practice)

(, 2548)






(, 2545)
(Wise Community Leaders)

51 | P a g e


1 1 ..2553

(, 2550)
4 1)
2)

3)

4)

(, 2545)
1)


2)


3)
1%



4)


5)
) )

52 | P a g e


1 1 ..2553

)
(, 2548)
(Science)


(Sustainable)
(, 2549)




( , 2550)


(,
2549)


(, 2551)
""

...
(wisdom) (knowledge)

...
...

53 | P a g e


1 1 ..2553

(holistic) ...
...




...

"
"




(, 2551)

(Participatory
Research) (Methodology)
(Model) (Social
Context)
54 | P a g e


1 1 ..2553

(Qualitative Research) (Case study)

(Documentary Study) (Field Notes)


1. (Documentary Study)
(Information Technology)


2. (Field Study)


2
2.1 (Observation)
(Survey)


1) (Participant Observation)


2) (Non-Participant Observation)


55 | P a g e


1 1 ..2553

2.2 (In-dept Interview)

(Non - Structural Interview) (Question)

()

(Key Information: KI)

2.3

5

3.





4.





56 | P a g e


1 1 ..2553

5.
(Descriptive Approach)

1 ..2519 6
500 13
5
30 502
() 75 2 /
4 13.50-15.50 . 4 34


17
..2523 4
211 (-) 15
57 3 40 27 415

( ) 3
70 68 79
4 39 09.30-11.30 . 13.00-15.00 .
3 3 13 1

57 | P a g e


1 1 ..2553

25
..2491 -
3 99 2,239 51
64
6 222
08.30-10.30 .

14 ..2534 270 9
65 5 24
332

67
15.30-16.30 .
14

1 () 5 ..2484
1,830 47
81 2 () 3
() 4 () 706/6 7



51

15.00-16.00 . 225

58 | P a g e


1 1 ..2553

30

4
-
1

2
(.)






3







59 | P a g e


1 1 ..2553

4 -
-




-
4
1

1 4 1
(Cycle) 1

(Dynamic)

(win-win)


60 | P a g e


1 1 ..2553

1.


2.

3.

. : . [Online] Available:
http://www.sufficiencyeconomy.org/show.php?id=11. [2008 , June 14].
, .. (2549). . :
.
. (2548, 4). . , 10.
. (2548, 18). . , 25.
. (2549). : . , 3 (8), 56-59.
. (2550). .
: .
. (2550, 12). . , 10.
. (2551). . :
.
. (2549). : . , 3 (8), 56-59.
, . (2545). . . :
.
. (2551, 21). . ., 9.

61 | P a g e


1 1 ..2553

. (2542). . 2,
: .
. (2545). . .:
.

62 | P a g e


1 1 ..2553

The study of Animal Sciences and Agricultural Technology Students' Satisfaction with StudentCentered Learning Activities for Agricultural Meteorology Course
1 1 2 1
3
1

3

2

63 | P a g e


1 1 ..2553

1 2551
16
(Geographic Information Systems
Laboratory) (Case Studies)

64 | P a g e


1 1 ..2553

Abstract
The purposes of this study was to investigate the Students' Satisfaction of Student-Centered Learning
activities in Agricultural Meteorology course offered in the first semester of the academic year 2008 for the
Animal Sciences and Agricultural Technology students included Geographic Information Systems Laboratory
and Case Studies. At the end of the course the satisfactions of the students to those activities were evaluated
by using students satisfaction questionnaires. Most students satisfied at a very satisfied level with those
activities because they got to explore more interesting knowledge.
Keyword: Satisfaction, Student-Centered Learning, Agricultural Meteorology

65 | P a g e


1 1 ..2553

..2542 4 22


24






30
(, 2550)

(712323
AGRICULTURAL METEOROLOGY)
( ..2549)
3 (3-0-6) 1
66 | P a g e


1 1 ..2553

3 (
, 2552)

(, 2550)
/

.. 2551

67 | P a g e


1 1 ..2553

(712323
AGRICULTURAL METEOROLOGY) 1 2551 15

5 3

(Likert) (Rating
scale) 5
5
4
3
68 | P a g e


1 1 ..2553

2
1

(2535:77)
4.50 - 5.00
3.50 - 4.49
2.50 - 3.49
1.50 - 2.49
1.00 - 1.49

5 Microsoft Excel

1.

69 | P a g e


1 1 ..2553

20
2. (Case Studies)

5
2 1-2
20


5

70 | P a g e


1 1 ..2553

4.20
4.33
4.27
4.40

3.87

4.27 1

71 | P a g e


1 1 ..2553

1 ()
5()

(40.00)

(26.67)

(20.00)

(6.67)

(6.67)

(0.00)

2.00

1.07

0.60

0.13

0.07

0.00

(33.33)

(33.33)

(20.00)

(6.67)

(6.67)

(0.00)

1.67

1.33

0.60

0.13

0.07

0.00

(53.33)

(33.33)

(6.67)

(0.00)

(6.67)

(0.00)

2.67

1.33

0.20

0.00

0.07

0.00

GIS

72 | P a g e


1 1 ..2553

3.87 3.80

73 | P a g e


1 1 ..2553

2 ()
5()

74 | P a g e

(40.00)

(26.67)

(20.00)

(6.67)

(6.67)

(0.00)

2.00

1.07

0.60

0.13

0.07

0.00

(40.00)

(33.33)

(26.67)

(0.00)

(0.00)

(0.00)

2.00

1.33

0.80

0.00

0.00

0.00

(53.33)

(26.67)

(13.33)

(6.67)

(0.00)

(0.00)

2.67

1.07

0.40

0.13

0.00

0.00

(33.33)

(26.67)

(26.67)

(6.67)

(6.67)

(0.00)

1.67

1.07

0.80

0.13

0.07

0.00

(26.67)

(26.67)

(26.67)

(6.67)

(13.33)

(0.00)

1.33

1.07

0.80

0.13

0.13

0.00

(33.33)

(40.00)

(20.00)

(6.67)

(0.00)

(0.00)

1.67

1.60

0.60

0.13

0.00

0.00

(53.33)

(33.33)

(13.33)

(0.00)

(0.00)

(0.00)

2.67

1.33

0.40

0.00

0.00

0.00


1 1 ..2553

1
2551

3.53

3.87
3.80

75 | P a g e


1 1 ..2553

100
(Self-Directed Learning)
(Learning resources)

1
2551

76 | P a g e


1 1 ..2553

/. ...
[online]
http://www.arcims.tmd.go.th/historyGIS.htm [20 2552].
. (2552). 2552
.(2550). 3
.
.
http://tlsd.rsu.ac.th/journal/Document/Journal105.pdf [21 2552]
. (2535). . :
.
. (2550).
[online]
http://www.mict.go.th/ewt_news.php?nid=1053&filename=index [20 2552].

77 | P a g e


1 1 ..2553

Effects of Dietary Protein and Energy Levels on Growth Performance in Growing Dairy Cattle
1 2
1

2

( 12 12 ) 24 (
144.111.0 . 91.2 ) randomized complete block design 4
3 X 2 factorial
3 6, 10 14 gCP/kgW0.75/d 2
792 885 kJ ME/kgW0.75/d 6

curvilinear
(10 gCP/kgW0.75) (885 kJ ME/kgW0.75)
710 //

: , , , ,

78 | P a g e


1 1 ..2553

Abstract
Twenty four growing Holstein crossbred dairy cattle (average weight 144.111.0 kg; average age
91.2 month) were used in a feeding trial to determine the effect of dietary energy and protein intake on feed
intake and weight gain and blood metabolites of the cattle fed whole sugarcane forage based diets. Treatments
were applied according to a 3 x 2 factorial arrangement in a randomized complete block design. Dietary
treatments were assigned to containing crude protein (CP) intake either 6, 10 or 14 gCP/kgW0.75/d. and
metabolizable energy (ME) intake either 792 or 885 kJ ME/kgW0.75/d. Higher dietary CP intake level
increased dry matter and organic matter intake. Average daily gain (ADG) was increased curvilinear by
dietary protein intake levels. In conclusion, dried whole sugarcane could be used as a roughage for growing
Holstein crossbred dairy cattle, and the optimum nutrient level was of protein (10 gCP/kgW0.75) and energy
(885 kJ ME/kgW0.75) for maximum weight gain of 710 gram per day and improvement growth performance.

Key Words: dairy cattle, energy, growth, protein, sugarcane forage

79 | P a g e


1 1 ..2553

70


(Saccharum officinarum)

(biomass) (Sommart et al.,
2005) Kawashima (2000)
27,219 10,886 8,500 MJ ME/ (2547)



(2547) 12

(NRC, 2001
and ARC, 1980)

(, 2547)
80 | P a g e


1 1 ..2553

( 12 12 )
24 ( 144.111.0 . 91.2 )

15 ECOMETIN - A,D3,E
Randomized complete block design (RCBD) 4
3 X 2 (Factorial)
3 6, 10 14 gCP/kgW0.75/d 2 792.45
884.92 kJ ME/kgW0.75/d 6
(1) - (2) - (3) - (4) -
(5) - (6) - (total mixed
ration, TMR) (dried whole sugar cane)
(Table 1)
42

07.00 16.30

(DM), (Ash), (CP) AOAC (1990)
(neutral detergent fiber, ADF),
(acid detergent fiber, NDF), (acid detergent fiber, ADL) Goering and Van
Soest (1970)

81 | P a g e


1 1 ..2553

(analysis of covariance) PROC GLM
SAS (SAS, 1996)
Orthogonal contrast Least square means
Least significant difference SAS
(SAS, 1996)

Table 2 (dry matter)
(organic matter) (P<0.01)
(2552) 5.00, 6.99,
8.95 10.62 gCP/kgW0.75 3.43, 4.98, 6.45 7.66
// Hoffman et al. (2001)
Lammers and Heinrichs (2000)
NRC
Bethard et al. (1997)
(rumen-undegradable)
Gabler and Heinrichs (2003a) (CP:ME)
quadratic

(neutral
detergent fiber, NDF) (acid detergent fiber,
ADF) (P<0.01) Broderick (2003)

82 | P a g e


1 1 ..2553

(interaction effect)
(P>0.05) (2552)

(15 gCP/kgW0.75) (1007 kJ ME/kgW0.75)

(average daily gain, ADG) (Table 2)


(P>0.05) Lammers and Heinrichs (2000)
(CP:ME)

(P<0.01)
MH (10 gCP/kgW0.75) (885 kJ ME/kgW0.75)
29.78 / 710.35
// HH (14
gCP/kgW0.75) (885 kJ ME/kgW0.75)


(2552) 5.00, 6.99, 8.95 10.62
gCP/kgW0.75 465, 876, 1399 1574 /
/ Brown et al. (2005)
2 14
Pirlo et al. (1997)
(110 %) NRC
83 | P a g e


1 1 ..2553

608.1 847.6 // Lammers and Heinrichs (2000)

NRC ( CP:ME
50:1 61.2:1 g/Mcal) 9
Schroeder and Titgemeyer (2008)

(2552)
(15 gCP/kgW0.75) (1007 kJ
ME/kgW0.75)

(10 gCP/kgW0.75) (885 kJ ME/kgW0.75)
710.35 //

(.) 2548

84 | P a g e


1 1 ..2553

. 2547. .
-
. , .
. 2547.
. -
. ,
.
. 2547.

. , .
. 2547. .
. , .
Takehiro Nishida . 2552 .
.
2552. .. 76-78.
Takehiro Nishida . 2552 .

. 2552.
.. 200-202.
ARC. 1965. The Nutrient Requirements of Farm Livestock. No. 2. Ruminants. Agric. Res. Counc., London,
UK.
AOAC. 1990. Official Methods of Analysis, Vol.1, 15th ed. Association of Official Analytical Chemists,
Washington, DC.
Bethard, G. L., R. E. James, and M. L. McGilliard. 1997. Effect of Rumen-Undegradable Protein and Energy
on Growth and Feed Efficiency of Growing Holstein Heifers. J. Dairy Sci. 80:2149-2155.

85 | P a g e


1 1 ..2553

Broderick, G. A. 2003. Effects of Varying Dietary Protein and Energy Levels on the Production of Lactating
Dairy Cows. J. Dairy Sci. 86:1370-1381.
Brown, E. G., M.J. VandeHaar, K. M. Daniels, J. S. Liesman, L. T. Chapin, D. H. Keisler, and M. S. Weber
Nielsen. 2005. Effect of increasing energy and protein intake on body growth and carcass composition
of heifer calves. J. Dairy Sci. 88:585594.
Gabler, M. T., and A. J. Heinrichs. 2003a. Dietary protein to metabolizable energy ratios on feed efficiency
and structural growth of prepubertal Holstein heifers. J. Dairy Sci. 86:268274.
Gabler, M. T., and A. J. Heinrichs. 2003b. Effects of increasing dietary protein on nutrient utilization in
heifers. J. Dairy Sci. 86:2170-2177.
Goering, H. K., and P. J. Van Soest. 1970. Forage fiber analyses. ARS Agric. Handbook No. 379. USDA,
Washington, DC.
Hoffman, P. C., N. M. Esser, L. M. Bauman, S. L. Denzine, M. Engstrom and H. Chester-Jones. 2001. Short
communication: Effect of dietary protein on growth and nitrogen balance heifers. J. Dairy Sci. 84: 843847.
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of energy and protein requirements for maintenance among Brahman cattle, swamp buffalo and Thai
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