• Title, Summary, Keyword: Cellulolytic Bacteria

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Isolation of Anaerobic Cellulolytic Bacteria from the Rumen of Holstein Dairy Cows to Develop Feed Additives for Ruminants (반추동물용 사료첨가제개발을 위한 홀스타인 젖소의 반추위로부터 분리한 혐기성 섬유소 분해균의 특성연구)

  • Choi, Nag-Jin;Lee, Gi-Young;Jeong, Kwang-Hwa;Kim, Chang-Hyun
    • 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.

Degradation of Rice Straw by Rumen Fungi and Cellulolytic Bacteria through Mono-, Co- or Sequential- Cultures

  • Ha, J.K.;Lee, S.S.;Kim, S.W.;Han, In K.;Ushida, K.;Cheng, K.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.6
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    • pp.797-802
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    • 2001
  • Two strains of rumen fungi (Piromyces rhizinflata B157, Orpinomyces joyonii SG4) and three strains of rumen cellulolytic bacteria (Ruminococcus albus B199, Ruminococcus flavefaciens FD1 and Fibrobacter succinogenes S85) were used as mono-cultures or combinationally arranged as co- and sequential-cultures to assess the relative contributions and interactions between rumen fungi and cellulolytic bacteria on rice straw degradation. The rates of dry matter degradation of co-cultures were similar to those of corresponding bacterial mono-cultures. Compared to corresponding sequential-cultures, the degradation of rice straw was reduced in all co-cultures (P<0.01). Regardless of the microbial species, the cellulolytic bacteria seemed to inhibit the degradation of rice straw by rumen fungi. The high efficiency of fungal cellulolysis seems to affect bacterial degradation rates.

Fermentation Property by Novel Cellulolytic Lactic Acid Bacteria Enterococcus sp. TO-94 on Omija (Schizandra chinensis Baillon) (신규 섬유질 분해성 젖산균 Enterococcus sp. TO-94를 이용한 오미자의 발효 특성)

  • Ryu, Il-Hwan;Lee, Eoh-Jin;Kwon, Ji-Wung;Lee, Kang-Soo;Kwon, Tae-Oh
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.6
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    • pp.429-438
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    • 2010
  • The use of cellulolytic lactic acid bacteria in new method to prepare high nutrition complementary foods was investigated. For the screening of cellulolytic lactic acid bacteria, more than 1,150 bacterial colony were isolated from diluted infant feces samples. A typical strain which appeared the most excellent cellulolytic activities was identified novel acidophilic Enterococcus sp. TO-94 through the results of morphological, biochemical and chemotaxonomic characteristics and 16S rDNA sequencing. The optimal lactic acid fermentation conditions of Omija(Schizandra chinensis Baillon) by Enterococcus sp. TO-94 were as follows: pH and temperature were 3.0 and $37^{\circ}C$, respectively, and fermentation time was 20hrs. The fructose and glucose were major free sugar and the contents were 5.83 and 4.30 mg/g after fermentation, respectively. The contents of lactic acid and acetic acid were 9.84 mg/g and 2.08 mg/g after fermentation, respectively. The vitamin $B_1$, $B_2$, niacin, folic acid and C were major vitamin in the fermented broth, the contents were 1.5~3 times higher than those of initial fermentation time. Also, the contents of polyphenol and anthocyanine were 3.8 and 1.2 times higher than those of initial fermentation time.

Effect of Roughage Sources on Cellulolytic Bacteria and Rumen Ecology of Beef Cattle

  • Wora-anu, S.;Wanapat, Metha;Wachirapakorn, C.;Nontaso, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.11
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    • pp.1705-1712
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    • 2007
  • The effect of different tropical feed sources on rumen ecology, cellulolytic bacteria, feed intake and digestibility of beef cattle was investigated. Four fistulated, castrated male crossbred cattle were randomly allocated to a $4{\times}4$ Latin square design. The treatments were: T1) urea-treated (5%) rice straw (UTS); T2) cassava hay (CH); T3) fresh cassava foliage (FCF); T4) UTS:FCF (1:1 dry matter basis). Animals were fed concentrates at 0.3% of body weight on a DM basis and their respective diets on an ad libitum basis. The experimental period was 21 days. The results revealed that the use of UTS, CH, FCF and UTS:FCF as roughage sources could provide effective fiber and maintain an optimal range of ruminal pH and $NH_3-N$. Total viable and cellulolytic bacterial populations were enhanced (p<0.05) with UTS as the roughage source. Animals fed FCF had a higher rumen propionate production (p<0.05) with a lower cellulolytic bacteria count. Moreover, three predominant cellulolytic bacteria species, namely Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens, were found in all treatment groups. Roughage intake and total DM intake were highest with UTS (2.2 and 2.5% BW, respectively) as the roughage source (p<0.05). Nutrient intake in terms of organic matter intake (OMI) was similar in UTS, CH and UTS:FCF treatments (8.0, 6.8 and 8.7 kg/d, respectively), while crude protein intake (CPI) was enhanced in CH, FCF and UTS:FCF as compared to the UTS treatment (p<0.05). Digestion coefficients of DM and organic matter (OM) were similar among treatments, while the CP digestion coefficients were similar in CH, FCF and UTS:FCF treatments, but were higher (p<0.05) in CH than in UTS. CP and ADF digestible intakes (kg/d) were highest (p<0.05) on the CH and UTS treatments, respectively. It was also observed that feeding FCF as a full-feed resulted in ataxia as well as frequent urination; therefore, FCF should only be fed fresh as part of the feed or be fed wilted. Hence, combined use of FCF and UTS as well as CH and FCF were recommended.

EFFECTS OF CHEMICAL TREATMENTS OF BARLEY STRAW ON LEACHING, AND DIGESTIBILITY BY RUMEN FLUID AND CELLULOLYTIC BACTERIA

  • Kudo, H.;Cheng, K.J.;Rode, L.M.;Abdullah, N.;Ho, Y.W.;Hussain, H.Y.;Jalaludin, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.3
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    • pp.389-396
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    • 1994
  • Effects of chemical treatments on in sacco and in vitro digestibility of barley straw by rumen fluid and pure cultures of cellulolytic bacteria were studied to evaluate the pretreatment and to improve the poor quality feed. Chemicals were applied by dissolving them in water equivalent to 40% of the weight of the straw (dry matter basis). Pretreatment with 5% NaOH yielded the largest increase in sacco digestion followed by pretreatment with 2% $(NH_4)_2SO_3$, 2.6% $NH_4OH$, 1.6% $NaHSO_3$ and untreated straw (control). In sacco dry matter digestibility of straw treated with NaOH and $(NH_4)_2SO_3$ continued to increase as the concentration of chemical increased (1 to 7.5%), as it was the in vitro dry matter loss by leaching. Treatment of barley straw with 5% NaOH enhanced significantly (p < 0.01) in vitro digestibility by rumen fluid, Fibrobacter suceinogenes and Ruminococcus albus though the fermentation products by cellulolytic bacteria were low, whereas the treatment with 5% $(NH_4)_2SO_3$ inhibited in vitro digestibility by F. succinogenes and R. albus together with lower fermentation products. Dry matter loss by leaching and bacterial digestion from barley straw treated with NaOH and $(NH_4)_2SO_3$ suggested the effect of pretreatment with these chemicals were based on leaching, and the cellulolytic bacteria had little to do with digestion.

Effects of Methylcellulose on Cellulolytic Bacteria Attachment and Rice Straw Degradation in the In vitro Rumen Fermentation

  • Sung, Ha Guyn;Kim, Min Ji;Upadhaya, Santi Devi;Ha, Jong K.;Lee, Sung Sill
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.9
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    • pp.1276-1281
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    • 2013
  • An in vitro experiment was conducted to evaluate the effect of methylcellulose on the attachment of major cellulolytic bacteria on rice straw and its digestibility. Rice straw was incubated with ruminal mixture with or without 0.1% methylcellulose (MC). The attachment of F. succinogenes, R. flavefaciens and R. albus populations on rice straw was measured using real-time PCR with specific primer sets. Methylcellulose at the level of 0.1% decreased the attachment of all three major cellulolytic bacteria. In particular, MC treatment reduced (p<0.05) attachment of F. succinogenes on rice straw after 10 min of incubation while a significant reduction (p<0.05) in attachment was not observed until 4 h incubation in the case of R. flavefaciens and R. albus. This result indicated F. succinogenes responded to MC more sensitively and earlier than R. flavefaciens and R. albus. Dry matter digestibility of rice straw was subsequently inhibited by 0.1% MC, and there was a significant difference between control and MC treatment (p<0.05). Incubated cultures containing MC had higher pH and lower gas production than controls. Current data clearly indicated that the attachment of F. succinogenes, R. flavefaciens and R. albus on rice straw was inhibited by MC, which apparently reduced rice straw digestion.

Studies on the Soil Microoganisms and Physiochemical Properties in Kwangju area (광주지역의 토양미생물과 물리화학적 특성에 관한 연구)

  • 김상문
    • Korean Journal of Plant Resources
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    • v.4 no.2
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    • pp.51-58
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    • 1991
  • With the soil samples collected from 33 locations in Kwangju area, the physiochemical properties of soil and soil microorganisms have been studied and the results of'the analysis were as follows ;1 . Tlle temperatilre, pH, moisture and organic matter of sampling s'tes were measured in the range of 21.O~28.O$^{\circ}C$, 4.0"6.6, 2.1"24.0% and 2.8~22.0% respectively.and that showed wide range distribution in moisture and organio mattrr particulary.and averase was 24.$^{\circ}C$ of temperature, 4.9 of pH, 11.9% of moisture and 8.9% of'organic matter.2. The general bacterial number, cellulolytic bacterial number, general fungal number and cellulolytic fungal number were measured in tile range of 23$\times$10$^{[-995]}$ _1548x10$^{[-995]}$ , 8.0$\times$10$^{[-995]}$ ~412.0$\times$10$^{[-995]}$ , 0.3$\times$10$^{[-994]}$ ~56.4$\times$10$^{[-994]}$ and 0.Ix10$^{[-994]}$ ~17.2x10$^{[-994]}$ , respectivelyand average was 378.4$\times$10$^{[-995]}$ of general bacteria, 102.5$\times$10$^{[-995]}$ of cellulolytic bacteria.13.OX10$^{[-994]}$ of general fungi and 4.3$\times$10$^{[-994]}$ of cellulolytic fungi.tic fungi.

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Characterization of Aerobic Cellulolytic Bacteria Part 1. Cultural Characteristics and Classification of Some Stock Cultures (섬유소 분해세균의 균학적 성질(제1보) -보존균주에 대한 배양적 성질 및 동정에 대하여-)

  • Choi, Woo-young
    • Korean Journal of Agricultural Science
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    • v.4 no.2
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    • pp.167-172
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    • 1977
  • Colony morphology, growth responses on some simple carbon sources and taxonomic position were established for three stock cultures of National Collection of Industrial Bacteria, Scotland. It was confirmed that NCIB 8077 belonged to the Cellulomonas species and that NCIB 8633 and NCIB 8634 belonged to the Pseudomonas species. Taxonomy of other cellulolytic bacteria published on various journals was also discussed.

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Effects of Mixtures of Tween80 and Cellulolytic Enzymes on Nutrient Digestion and Cellulolytic Bacterial Adhesion

  • Hwang, Il Hwan;Lee, Chan Hee;Kim, Seon Woo;Sung, Ha Guyn;Lee, Se Young;Lee, Sung Sill;Hong, Hee Ok;Kwak, Yong-Chul;Ha, Jong K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.11
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    • pp.1604-1609
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    • 2008
  • A series of in vitro and in vivo experiments were conducted to investigate the effects of the mixture of Tween 80 and cellulolytic enzymes (xylanase and cellulase) on total tract nutrient digestibility and rumen cellulolytic bacterial adhesion rates in Holstein steers. Ground timothy hay sprayed with various levels of Tween 80 and cellulolytic enzymes was used as substrates in an in vitro experiment to find out the best combinations for DM degradation. The application level of 2.5% (v/w) Tween 80 and the combination of 5 U xylanase and 2.5 U cellulase per gram of ground timothy hay (DM basis) resulted in the highest in vitro dry matter degradation rate (p<0.05). Feeding the same timothy hay to Holstein steers also improved in vivo nutrient (DM, CP, CF, NDF and ADF) digesibilities compared to non-treated hay (p<0.05). Moreover, Tween 80 and enzyme combination treatment increased total ruminal VFA and concentrations of propionic acid and isovaleric acid with decreased acetate to propionate ratio (p<0.001). However, adhesion rates of Fibrobacter succinogenes and Ruminococcus flavefaciens determined by Real Time PCR were not influenced by the treatment while that of Ruminococcus albus was decreased (p<0.05). The present results indicate that a mixture of Tween 80 and cellulolytic enzymes can improve rumen environment and feed digestibility with variable influence on cellulolytic bacterial adhesion on feed.