• Asian-Australasian Journal of Animal Sciences
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• v.23 no.8
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• pp.1022-1027
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• 2010

Effects of increasing dietary rice straw on chewing activity, ruminal fermentation, and fibrolytic enzyme activity in growing goats were investigated in a $4{\times}4$ Latin Square experiment. The goats were offered four diets with an increasing proportion of rice straw (i.e. 0.05, 0.10, 0.15 and 0.20, respectively, on dry matter basis). Increasing level of rice straw increased ($P_{linear\;effect}$ <0.05) the time spent on eating, ruminating, and chewing. The ruminal pH and acetate: propionate ratio were increased ($P_{linear\;effect}$ <0.05), while the $NH_3$-N concentration was decreased ($P_{linear\;effect}$ <0.01). Increasing level of rice straw in the diet increased ($P_{linear\;effect}{\leq}0.01$) molar proportion of acetate and isovalerate, and decreased ($P_{linear\;effect}$ <0.01) molar proportion of propionate. The CMCase, xylanase and cellobiase activities in the rumen were decreased ($P_{linear\;effect}$ <0.05) with increasing level of dietary rice straw, whereas the avicelase activity was increased ($P_{linear\;effect}$ <0.01). In summary, increased level of rice straw elevated the dietary neutral detergent fibre (NDF) content in the diet and had a great impact on chewing activity and ruminal fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.8
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• pp.1110-1117
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• 2001

Responses of the rumen fungus, Neocallimastix frontalis RE1, to long chain fatty acid (LCFA) were evaluated by measuring gas production, filter paper (FP) cellulose digestion and polysaccharidase enzyme activities. LCFA (stearic acid, $C_{18:0}$; oleic acid, $C_{18:1}$; linoleic acid, $C_{18:2}$ and linolenic acid, $C_{18:3}$) were emulsitied by ultrasonication under anaerobic condition, and added to the medium. When N frontalis RE1 was grown in culture with stearic, oleic and linoleic acid, the cumulative gas production, gas pool size, FP cellulose digestion and enzymes activities significantly (p<0.05) increased at some incubation times(especially, exponential phases of fungal growth, 48~120 h of incubation) relative to that for control cultures. However, the addition of linolenic acid strongly inhibited all of the investigated parameters up to 120 h incubation, but not after 168 and 216 h of incubation. These results indicated that stearic, oleic and linoleic acids tended to have great stimulatory effects on fungal cellulolysis, whereas linolenic acid caused a significant (p<0.05) inhibitory effects on the cellulolysis by the rumen fungus. These results are the first report of the effect of LCFAs on the ruminal fungi. Further research is needed to identify the mode of action of LCFAs on fungal strains and to verify whether or not ruminal fungi have ability to hydrate unsaturated LCFAs to saturated FAs. There was high correlation between cumulative in vitro gas production and fungal growth (94.78%), FP cellulose degradation (96.34%), CMCase activity(90.86%) or xylanase activity (87.67%). Thus measuring of cumulative gas production could be a useful tool for evaluating fungal growth and/or enzyme production by ruminal fungi.

• Korean Journal of Soil Science and Fertilizer
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• v.34 no.5
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• pp.333-341
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• 2001

The Compost-decomposing-bacteria was isolated from livestock compost containing sawdust. The isolated bacteria was identified as Bacillus subtilis LYH201 by the method of the composition of the fatty acid with MIDI system and Bergey's manual. Cloning of CMCase encoding gene was accompanied by shotgun method. The pLK100 have yellow activity ring on CMC medium, that was carried 2.2 kb insert DNA in pBluescript II $SK^+$ vector, named BglC gene. The BglC was very similar to Pectobacterium carotovorum Gun_CLOAB(P15704) with score of 57% identity and 71% homology over 508 aa. The BglC was measured molecular weight 56 kDa by CMC-SDS-PAGE. Optimum cellulase activity Bacillus subtilis LYH201 was temperature $50^{\circ}C$ and pH 7.

• Journal of Life Science
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• v.17 no.12
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• pp.1660-1668
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• 2007

The non-ionic surfactant (NIS) Tween 80 was evaluated for its ability to influence invitro cumulative gas production, dry matter digestibility, cellulolytic enzyme activities, anaerobic microbial growth rates, and adhesion to substrates by mixed rumen microorganisms on rice straw, alfalfa hay, cellulose filter paper and tall fescue hay. The addition of NIS Tween 80 at a level of 0.05% increased significantly (P<0.05) in vitro DM digestibility, cumulative gas production, microbial growth rate and cellulolytic enzyme activity from all of substrates used in this study. In vitro cumulative gas production from the NIS-treated substrates; rice straw, alfalfa hay, filter paper and tall fescue hay was significantly (P<0.05) improved by 274.8, 235.2, 231.1 and 719.5% compared with the control, when substrates were incubated for 48 hr in vitro. The addition of 0.05% NIS Tween 80 to cultures growing on alfalfa hay resulted in a significant increase in CMCase (38.1%), xylanase (121.4%), Avicelase (not changed) and amylase (38.2%) activities after 36 h incubation. These results indicated that the addition of 0.05% Tween 80 could greatly stimulate the release of some kinds of cellulolytic enzymes without decreasing cell growth rate in contrast to trends reported with aerobic microorganism. Our SEM observation showed that NIS Tween. 80 did not influence the microbial adhesion to substrates used in the study. Present data clearly show that improved gas production, DM digestibility and cellulolytic enzyme activity by Tween 80 is not due to increased bacterial adhesion on the substrates.

• Korean Journal of Microbiology
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• v.41 no.1
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• pp.81-86
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• 2005

One of the endoglucanases, F-I-III, was purified from the culture filtrate of T. sp. C-4 through procedures including chromatography on Sephacryl S-200, DEAE-Sepharose A-50, and Chromatofocusing on Mono-P (FPLC). The molecular weight of the enzyme was determined to be about 56,000 Da by SDS-PAGE, and pI of 4.9 by analytical isoelectric focusing. F-I-III showed the highest enzyme activity at $55^{\circ}C$, and the pH optimum of the enzyme was 5.0. There was no loss of activity when the enzyme was incubated at $50^{\circ}C$ for 24 hours. The specific activity of the enzyme F-I-III toward the CMC was 315.4 U/mg. The Km value for $PNPG_2$ of F-I-III was 2.69 mM. N-terminal sequence of F-I-III was analyzed to be QPGTSTPEVHPKKLTTYK. It showed $95\%$ of homology to that of EGI from T. reesei. The presence of some metal ions (1 mM) had only a little effect on CMCase activity. The treatment of the reducing agents resulted in the increase of endoglucanase activity.

• Korean Journal of Agricultural Science
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• v.25 no.1
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• pp.131-137
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• 1998

The cellulase components were partially purified from the culture filtrate of the alkalophilic bacterium Pseudomonas sp. AC-711 and its enzymatic properties were characterized. The specific activity of the purified major enzyme component was 3.5 units/mg protein as carboxymethyl cellulase and the yield was 23% of the total activity of the culture broth. The molecular weight of the component was 46,000 and the Km and Vmax on CMC were determined as $15.4mg\;mL^{-1}$ and $4.17{\mu}moles\;mL^{-1}\;min^{-1}$, respectively. The enzyme was stable at the temperatures below $60^{\circ}C$ and at the pH range of 4.0~11.0, and the optimal temperature and pH were $60^{\circ}C$ and pH 8.0, respectively. The enzyme activity was not significantly affected by the common surfactants (concentration: 0.05%) such as ${\alpha}$-olefin sulfonate, linear alkylbenzene sulfonate, sodium dodecyl sulfonate, hexadecyltrimethylammonium bromide and Tween 80. The enzyme was activated by the metal ions such as $Ca^{2+}$, $Cu^{2+}$, $Co^{2+}$, whereas inhibited by $Hg^{2+}$ and $Zn^{2+}$. The enzyme exhibited relatively high activity toward amorphous CMC as compared with crystalline substrates such as filter paper and avicel.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.10
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• pp.1435-1439
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• 2005

Bacterio-mineral water (BMW) produced from manure has been known to exert a number of positive effects on animal production and odor control. An experiment was conducted to examine the effects of BMW produced from bio-reacted swine manure on in vitro gas production, cellulose degradation, microbial growth and fibrolytic enzyme activities of mixed rumen microorganisms. The five levels of 0, 0.001, 0.005, 0.01 and 1.0% BMW were supplemented into serum vials containing mixed rumen microorganisms. Incubations were carried out anaerobically at $39^{\circ}C$ without shaking for 0, 12, 24, 48, 72 and 96 h. There were no significant (p>0.05) differences among the treatments for the initial rate of gas production. At 72 h incubation, the gas production tended (p<0.1) to be increased by the 0.01 and 1.0% BMW treatments compared with control and the 0.001% BMW treatment. At the end of incubation (96 h), the sample supplemented with 0.01% BMW was higher (p<0.05) than control (0% BMW) in the gas production. The microbial growth rate was increased by all the BMW treatments, while 0.01% BMW was most effective in stimulating the growth rate. Although the addition of BMW on the filter paper DM degradation was not significantly influenced throughout the incubation period except the 48 h incubation, DM degradation tended to be increased by all BMW treatments compared with control. The addition of both 0.005 and 0.01% BMW highly increased (p<0.05) CMCase activity compared with control after 24 h and 48 h incubation, while at the 72 h incubation the 0.01% BMW addition only significantly increased (p<0.05). After 72 h incubation, the xylanase activity was significantly (p<0.05) increased with the addition of 1.0% BMW compared with the addition of 0.001 and 0.005% BMW, while at the other incubation times, the xylanase activity was not different among the treatments. In conclusion, the 0.01% BMW of supplementation level would be the suitable addition level to stimulate rumen fermentation increasing microbial growth and cellulose degradation.

• Korean Journal of Soil Science and Fertilizer
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• v.34 no.6
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• pp.394-400
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• 2001

This experiment was conducted to determine the safety and feasibility of using compost-promoting-bacteria. Compost-promoting-bacteria was isolated from livestock compost containing sawdust. The isolated bacteria was identified as Bacillus subtilis LYH201 by the method of the composition of the fatty acid with MIDI system and Bergey's manual. This Bacillus subtilis LYH201 had the following characteristics : Gram-positive, straight rod ($0.5{\sim}0.7{\mu}m$ width, $2.5{\sim}3.0{\mu}m$ length), facultatively aerobic and product of xylanase, CMCase, catalase, oxidase, protease and $0.5{\sim}0.7{\mu}m$-amylase. Growth of Bacillus subtilis LYH201 at saccharose as carbon source(0.5%) was faster than other carbon source. Activity of cellulase. $0.5{\sim}0.7{\mu}m$-amylase and protease from Bacillus subtilis LYH201 after 24 hours at $50^{\circ}C$ by agar diffusion method was higher than that of low temperature. Optimum growth condition of Bacillus subtilis LYH201 was $50^{\circ}C$ and pH 6.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.3
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• pp.337-342
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• 2004

A non-ionic surfactant, Tween 80 has been known to exert a number of positive effects on degradative enzymes in in vitro aerobic and anaerobic microbial cultures. An experiment was conducted to examine effects of supplementation of Tween 80 on ruminal fermentation and nutrient digestibility of Hanwoo steers. The experiment was designed as a $3{\times}3$ Latin square with duplication and six Hanwoo steers with rumen cannulae, average weight 497 (SE 61.1) kg. For the experiment the animals were given a basal diet consisting of rice straw and compound feed mixed at 4:6 ratio. The three experimental treatments were (1) the basal diet, supplemented with (2) 5 g/d Tween 80 and (3) 10 g/d Tween 80. Ruminal pH was significantly (p<0.05) affected by Tween 80 supplementation at 6 h after feeding. Increasing supplementation levels of Tween 80 linearly increased the total VFA concentration. CMCase activity by the 10g/d supplementation of Tween 80 were significantly increased (p<0.05) by 24.4% compared with that of control. Digestibility of crude fiber was significantly increased (p<0.05) in Hanwoo steers fed the diet supplemented with 10 g/d Tween 80 compared with those of control, whilst digestibility of ether extract (EE) was linearly increased by increasing Tween 80 supplementation level (p<0.05). In other nutrient components, their digestibilities of Hanwoo fed diets supplemented with Tween 80 tended to increase. It is concluded that Tween 80 has a potential for industrial application as a feed additive to improve ruminant production.

• Korean Journal of Organic Agriculture
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• v.20 no.3
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• pp.327-343
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• 2012

In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.