• Asian-Australasian Journal of Animal Sciences
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• v.33 no.10
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• pp.1590-1598
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• 2020

Objective: The objective of this study was to evaluate the effects of lysophospholipids (LPL) supplementation on rumen fermentation, degradability, and microbial diversity in forage with high oil diet in an in vitro system. Methods: Four experimental treatments were used: i) annual ryegrass (CON), ii) 93% annual ryegrass +7% corn oil on a dry matter (DM) basis (OiL), iii) OiL with a low level (0.08% of dietary DM) of LPL (LLPL), and iv) OiL with a high level (0.16% of dietary DM) of LPL (HLPL). An in vitro fermentation experiment was performed using strained rumen fluid for 48 h incubations. In vitro DM degradability (IVDMD), in vitro neutral detergent fiber degradability, pH, ammonia nitrogen (NH₃-N), volatile fatty acid (VFA), and microbial diversity were estimated. Results: There was no significant change in IVDMD, pH, NH₃-N, and total VFA production among treatments. The LPL supplementation significantly increased the proportion of butyrate and valerate (Linear effect [Lin], p = 0.004 and <0.001, respectively). The LPL supplementation tended to increase the total bacteria in a linear manner (p = 0.089). There were significant decreases in the relative proportions of cellulolytic (Fibrobacter succinogenes and Ruminococcus albus) and lipolytic (Anaerovibrio lipolytica and Butyrivibrio proteoclasticus) bacteria with increasing levels of LPL supplementation (Lin, p = 0.028, 0.006, 0.003, and 0.003, respectively). Conclusion: The LPL supplementation had antimicrobial effects on several cellulolytic and lipolytic bacteria, with no significant difference in nutrient degradability (DM and neutral detergent fiber) and general bacterial counts, suggesting that LPL supplementation might increase the enzymatic activity of rumen bacteria. Therefore, LPL supplementation may be more effective as an antimicrobial agent rather than as an emulsifier in the rumen.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.209-220
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• 2015

In this study, gut bacterial communities in xylophagous insects were analyzed using the pyrosequencing of 16S rRNA genes for their potential biotechnological applications in lignocelluloses degradation. The result showed that operational taxonomic units (OTUs), species richness and diversity index were higher in the hindgut than in the midgut of all insect samples analyzed. The dominant phyla or classes were Firmicutes (54.0%), Bacteroidetes (14.5%), ${\gamma}-Proteobacteria$ (12.3%) in all xylophagous insects except for Rhinotermitidae. The principal coordinates analysis (PCoA) showed that the bacterial community structure mostly clustered according to phylogeny of hosts rather than their habitats. In our study, the two carboxymethyl cellulose (CMC)-degrading isolates which showed the highest enzyme activity were most closely related to Bacillus toyonensis $BCT-7112^T$ and Lactococcus lactis subsp. hordniae $NCDO\;2181^T$, respectively. Cellulolytic enzyme activity analysis showed that ${\beta}-1,4-glucosidase$, ${\beta}-1,4-endoglucanase$ and ${\beta}-1,4-xylanase$ were higher in the hindgut of Cerambycidae. The results demonstrate that xylophagous insect guts harbor diverse gut bacteria, including valuable cellulolytic bacteria, which could be used for various biotechnological applications.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.10
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• pp.1429-1433
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• 2001

This study provides electron microscopic evidence for the presence of cellulosome-like structures on the cell surface of Ruminococcus albus F-40. Electron microscopy showed that clusters of tightly packed spherical particles were located on the cell surface of R. albus. The protuberant structures present mainly on the bacterial surface and also bound to the cellulose substrate appeared to be the site of cellulosome-like structures. From the evidence presented, we suggest that the structures described here might be a characteristic feature of some ruminal cellulolytic bacteria.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1979.04a
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• pp.114.2-115
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• 1979

Horikoshi등이 호 alkali 성 미생물에 관한 연구에서 분리한 수종의 cellulolytic bacteria중에서 가장 강력한 균체의 효소를 생산하는 Aeromonas 속 세균의 cellulolytic 효소에 관한 연구 결과를 보고한다. 공업적으로 생산된 효소를 사용하여 효소작용의 최적조건을 측정하고 gel filtration, ion-exchange chromatography 및 affinity chromatography 로 cel-luplytic 효소를 분리정제하였다. 본 효소의 활성 최적 pH는 7.0~8.5로 alkaline 효소였으며 반응온도 5$0^{\circ}C$에서 가장 강한 활성을 보였다. 분리 정제과정에서 carboxymethyl cellulose (CMC)에 대하여 활성이 있는 단백질이 최소 8종이상 분리되었으며 이중 1개 효소는 CMC에 대해서는 극히 낮은 활성을 보였으나 결정성 기질인 Avicel 에는 강한 활성을 보였다. 본 연구의 결과를 Cellulomonas속 세균 및 Trichoderma속 곰팡이의 효소와 그 성질을 비교 검토하였다.

• Journal of Microbiology and Biotechnology
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• v.27 no.3
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• pp.573-583
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• 2017

Biofilm formation on the membrane surface results in the loss of permeability in membrane bioreactors (MBRs) for wastewater treatment. Studies have revealed that cellulose is not only produced by a number of bacterial species but also plays a key role during formation of their biofilm. Hence, in this study, cellulase was introduced to a MBR as a cellulose-induced biofilm control strategy. For practical application of cellulase to MBR, a cellulolytic (i.e., cellulase-producing) bacterium, Undibacterium sp. DM-1, was isolated from a lab-scale MBR for wastewater treatment. Prior to its application to MBR, it was confirmed that the cell-free supernatant of DM-1 was capable of inhibiting biofilm formation and of detaching the mature biofilm of activated sludge and cellulose-producing bacteria. This suggested that cellulase could be an effective anti-biofouling agent for MBRs used in wastewater treatment. Undibacterium sp. DM-1-entrapping beads (i.e., cellulolytic-beads) were applied to a continuous MBR to mitigate membrane biofouling 2.2-fold, compared with an MBR with vacant-beads as a control. Subsequent analysis of the cellulose content in the biofilm formed on the membrane surface revealed that this mitigation was associated with an approximately 30% reduction in cellulose by cellulolytic-beads in MBR.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.9
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• pp.1397-1401
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• 2007

An in situ experiment was conducted to find out whether Tween 80 improves rice straw digestion through increased adhesion of major fibrolytic bacteria. Rice straw was sprayed with various levels of Tween 80 non-ionic surfactant or SDS ionic surfactant 24 h before incubation in the rumen of Holstein steers. Dry matter (DM) disappearance and adhesion of F. succinogenes, R. flavefaciens and R. albus on rice straw after in situ incubation were measured by real-time PCR. Application of Tween 80 increased DM disappearance, which was more noticeable at an application level of 1% compared to lower application levels. Application of SDS resulted in an opposite response in DM disappearance with highest reduction in DM disappearance at 1% level. In a subsequent in situ experiment, higher Tween 80 was applied to rice straw in an attempt to find the optimum application level. Tween 80 at 2.5% gave better DM disappearance than 1% with a similar result at 5%. Therefore, an adhesion study was carried out using rice straw treated with 2.5% Tween 80. Our results indicated that Tween 80 reduced adhesion of all three major rumen fibrolytic bacteria to rice straw. Present data clearly show that improved DM disappearance by Tween 80 is not due to increased bacterial adhesion onto substrates.

• The Korean Journal of Mycology
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• v.30 no.2
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• pp.147-151
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• 2002

Trichoderma spp. are the aggressive causal agents for green mold disease on oyster mushroom (Pleurotus spp.) cultivation. Antifungal bacteria (KATB 99121, KATB 99122 and KATB 99123 strains) were isolated from the compost for Pleurotus ostreatus. Among these bacterial strains, KATB 99121 strain showed an excellent inhibitory activity to the pathogens for green molds such as T. harzianum, T. viride and T. hamatum and an animal pathogen, Candida albicans, but did not affect on the culture of Pleurotus ostreatus (2209, Chunchu 2 and Wonhyung strains). KATB 99121 strain secreted amylolytic, proteolytic and cellulolytic exoenzymes. KATB 99122 and KATB 99123 strains excreted amylolytic, proteolytic, cellulolytic, lipolytic exoenzymes and showed ${\beta}$-glucosidase activity. Further studies will be conducted on the development of microbial fungicides using the antagonistic bacteria for the control of green mold disease on Pleurotus spp.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.5
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• pp.672-676
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• 2003

Effect of a surfactant Tween 80 on the bacterial growth in the rumen was examined on the in vitro pure cultures of Streptococcus bovis, Selenomonas ruminantium, Butyrivibrio fibrisolvens, Prevotella ruminicola, Megasphaera elsidenni, Fibrobacta succinogenes, Ruminanococcus albus and Ruminococcus flavefaciens. Dry matter degradability (DMD), concentrations and compositions of volatile fatty acids (VFA), and the most probable number (MPN) of cellulolytic bacteria and total number of bacteria in the presence of Tween 80 were also examined on the in vitro rumen mixed culture either with barley grain or orchardgrass hay. The growth of S. bovis, S. ruminantium, B. fibrisolvens, P. ruminicola, M. elsidenni and F. succinogenes were significantly higher (p<0.05) at over 0.05% concentrations of Tween 80 than those of the control cultures, while was not changed with R. albus and R. flavefaciens. With rumen mixed culture the DMD of barley grain and orchardgrass hay was significantly higher (p<0.05) at a 0.2% concentration of Tween 80 than the control, being reflected in the significantly higher (p<0.05) VFA production (mmol $g^{-1}$DDM) with orchardgrass hay. The higher (p<0.05) ratio of propionate to acetate at a 0.2% concentration of Tween 80 was also observed with orchardgrass hay, showing a similar trend with barley grain. No changes in the total bacterial number and MPN of cellulolytic bacteria were observed.

• Microbiology and Biotechnology Letters
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• v.50 no.2
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• pp.245-254
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• 2022

Cellulases are a group of biocatalyst enzymes that are capable of degrading cellulosic biomass present in the natural environment and produced by a large number of microorganisms, including bacteria and fungi, etc. In the current study, we isolated, screened and characterized cellulase-producing bacteria from soil. Three cellulose-degrading species were isolated based on clear zone using Congo red stain on carboxymethyl cellulose (CMC) agar plates. These bacterial isolates, named as HB2, HS5 and HS9, were subsequently characterized by morphological and biochemical tests as well as 16S rRNA gene sequencing. Based on 16S rRNA analysis, the bacterial isolates were identified as Bacillus cerus, Bacillus subtilis and Bacillus stratosphericus. Moreover, for maximum cellulase production, different growth parameters were optimized. Maximum optical density for growth was also noted at pH 7.0 for 48 h for all three isolates. Optical density was high for all three isolates using meat extract as a nitrogen source for 48 h. The pH profile of all three strains was quite similar but the maximum enzyme activity was observed at pH 7.0. Maximum cellulase production by all three bacterial isolates was noted when using lactose as a carbon rather than nitrogen and peptone. Further studies are needed for identification of new isolates in this region having maximum cellulolytic activity. Our findings indicate that this enzyme has various potential industrial applications.

• Journal of Animal Science and Technology
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• v.51 no.1
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• pp.45-52
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• 2009

This experiment was conducted to observe the effects of anaerobic cellulolytic bacteria culture (Ruminococcus flavefaciens H-20 and Fibrobactor succinogenes H-23) on in vivo ruminal fermentation characteristics in Hanwoo heifers. Four ruminally cannulated Hanwoo heifers ($221\pm7.5kg$) receiving a basal diet containing 3 kg of mixture hay (tall fescue and ochardgrass) and 2 kg of concentrate per day were in a $4\times4$ Latin square with 21-day periods. Treatments were the basal diet without the culture additive (control), the basal diet plus 50 ml/day of bacteria culture of H-20 and H-23 (1%), 150 ml/day of H-20 and H-23 (3%), and 250 ml/day of H-20 and H-23 (5%). In the whole experimental periods, ruminal pH did not differ between treatments. However, the concentration of ruminal ammonia-N was increased in the 3% treatment relative to control and the 1% treatment at 1 hr post-feeding (p<0.05). Avicelase and CMCase (carboxymethyl cellulase) activities in rumen fluid showed no significant difference among treatments. However, xylanase activity was higher in the 5% (119.49, xylose ${\mu}mol$/ml/min) than the 3% treatment (71.02, xylose ${\mu}mol$/ml/min) at 0 hr post-feeding (p<0.05). Concentrations of ruminal total VFA, acetate, propionate and valerate were unaffected by treatments, while butyrate was higher in the 3% treatment (24.48 mM) than control (15.71 mM) at 1 hr post-feeding (p<0.05). Results indicate that minimum 3% inclusion of cellulolytic bacteria cultures improved ruminal fermentation, especially ammonia-N concentration and butyric acid production.