• Journal of Animal Science and Technology
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• 제52권5호
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• pp.413-426
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• 2010

본 연구는 반추가축에 급여되는 사료원 중 종류별, 수준별 lignosulfonate 처리 대두박의 반추위 내 undegradable protein (UDP) 효율을 알아보기 위하여 실험 I 에서 전체 4종의 lignosulfonate (Desulfonated, Na, Ca, Solution)를 각각 수준별 (2, 4, 8%)로 처리하여 전체 12종의 lignosulfoanted soybeanmeal (LSBM)에 대한 반추위 내 미생물 발효 특성의 미치는 영향을 알아보기 위해 in vitro 시험이 실시되었다. 실험 H에서는 실험 I 에서 대두박의 단백질 보호 효과가 가장 높은 LSBM Na 2% 처리구와 LSBM Solution (LSBM Sol) 2% 처리구 그리고 열처리 LSBM (LSBM Heat) 2%를 이용하여 rumen simulation continuous culture (RSCC) system을 이용한 반추위 내 미생물 발효 특성에 대한 연구가 실시되었다. 시험 I in vitro 시험에서 모든 LSBM 처리구에서 전체 배양시간 평균 pH수준, gas 생성량, $NH_3$-N 농도, 건물소화율, VFA 생성량 그리고 ADF 소화율은 배양시간이 지남에 따라 대조구에 비해 낮은 결과를 나타내었다(p<0.05). 반추위 내 미생물단백질 합성량은 대조구가 LSMB 처리구 보다 통계적으로 높은 결과를 나타내었으며 (p<0.05), LSBM 수준이 증가할수록 반추위 내 미생물 단백질 합성량은 감소하는 경향을 나타내었다. UDP 함량은 대조구보다 LSBM 처리구에서 높은 결과를 나타내었으며(p<0.05), 특히 LSBM Sol 처리구는 대조구에 비해 각각 2%, 4% 그리고 8% 처리군에서 11.18%, 11.54% 그리고 29.11%의 UDP 함량이 증가 하였다. 시험 II는 RSCC system을 이용하여 반추위 내 미생물 발효특성에 대한 시험이 실시되었다. 시험사료는 대조구와 In vitro 시험 중 대조구와 UDP 효과가 높은 LSBM Na 2%, LSBM Sol 2% 그리고 LSBM Heat 2% 처리한 사료 4처리 4반복($4{\times}4$ Latin squire)에 의해 15일간 시험이 실시되었으며, 배양 마지막 3일 동안의 sample을 이용하여 분석을 실시하였다. 전체 배양 기간 중 모든 처리구에서 적응기간 동안 불규칙적인 반추위 미생물 발효 특성을 보였으나 배양 12일 이후 안정된 발효특성을 나타내었다. Sampling 기간 동안의 각 처리간의 평균 pH는 모든 LSBM 처리구에서 대조구보다 높았다(p<0.05). $NH_3$-N 함량, VFA 함량 그리고 반추위 내 미생물 단백질 합성량은 LSBM 처리구가 대조구에 비해 낮은 결과를 나타내었다(p<0.05). UDP 함량은 대조구, LSBM Heat 2%, LSBM Na 2% 그리고 LSBM Sol 2%에서 각 41.55, 43.49, 45.28 그리고 43.52%로 모든 LSBM 처리구가 대조구에 비해 유의하게 높은 경향을 나타내였다(p<0.05). 시험 I과 시험II에서 UDP 함량은 LSBM 처리구에서 lignosulfonate 처리 수준이 증가할수록 증가하는 경향을 나타내었고, 모든 LSBM 처리구는 대조구에 비해 높은 UDP 함량을 나타내었다. 이와 같은 결과는 사료원으로써 대두박에 대한 lignosulfonate 처리가 coating 작용과 열처리에 의한 물리 화학적 효과로 인하여 대두박의 단백질 분해율을 저해 시킨 것으로 사료된다. 그러나 LSBM 처리군 중 건물소화율과 반추위 내 미생물단백질 합성량은 LSBM 2% 처리구에서 가장 높은 결과를 나타내였다. 따라서, 반추가축 사료내 LSBM 첨가가 In vitro와 RSCC system을 이용한 반추위내 미생물발효 특성 시험에서 반추위 내에서 by-pass protein 함량을 증가시키는 결과를 나타내었으며, 추후 착유우를 이용한 LSBM 급여를 시험을 통하여 유단백질 증진효과에 대하여 추가적인 평가가 이루어져야 할 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제29권3호
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• pp.365-371
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• 2016

This study was aimed to evaluate the stability of conjugated linoleic acids (CLAs) by nano-encapsulation against in vitro ruminal biohydrogenation by microbial enzymatic conversion. CLAs (free fatty acid form of CLA [CLA-FFA], nano-encapsulated CLA-FFA, triglyceride form of CLA [CLA-TG], and nano-encapsulated CLA-TG) were used in the in vitro fermentation experiments. When Butyrivibrio fibrisolvens (B. fibrisolvens) was incubated with CLA-FFAs, the concentrations of cis-9, trans-11 CLA and vaccenic acid (VA) slightly was decreased and increased by nano-encapsulation, respectively. When B. fibrisolvens was incubated with CLA-TG, the concentrations of cis-9, trans-11 CLA and VA decreased, but these were increased when B. fibrisolvens was incubated with nano-encapsulated CLA-TG. The nano-encapsulation was more effective against the in vitro biohydrogenation activity of B.fibrisolvens incubated with CLA-FFA than with CLA-TG. In the in vitro ruminal incubation test, the total gas production and concentration of total volatile fatty acids incubated with nano-encapsulated CLA-FFA and CLA-TG were increased significantly after 24 h incubation (p<0.05). Nano-encapsulated CLA-FFA might, thus, improve the ruminal fermentation characteristics without adverse effects on the incubation process. In addition, nano-encapsulated CLA-FFA increased the population of Fibrobacter succinogenes and decreased the population of B. fibrisolvens population. These results indicate that nano-encapsulation could be applied to enhance CLA levels in ruminants by increasing the stability of CLA without causing adverse effects on ruminal fermentation.

• Asian-Australasian Journal of Animal Sciences
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• 제19권12호
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• pp.1761-1768
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• 2006

In this study, the comparative effects of transgenic corn (Mon 810 and Event 176) and isogenic corn (DK729) were investigated for their influence on in vitro rumen fermentation. This study consisted of three treatments with 0.25 g rice straw, 0.25 g of corn (Mon810/Event176/DK 729) mixed with 30 ml rumen fluid-basal medium in a serum bottle. They were prepared in oxygen free conditions and incubated at $39^{\circ}C$ in a shaking incubator. The influence of transgenic corn on the number of bacterial population, F. succinogenes (cellulolytic) and S. bovis (amylolytic), was quantified using RT-PCR. Fermentative parameters were measured at 0, 2, 4, 8, 12 and 24 h and substrate digestibility was measured at 12 and 24 h. No significant differences were observed in digestibility of dry matter, NDF, ADF at 12 and 24 h for both transgenic and isogenic form of corns (p>0.05) as well as in fermentative parameters. Fluid pH remained unaffected by hybrid trait and decreased with VFA accumulation as incubation time progressed. No influence of corn trait itself was seen on concentration of total VFA, acetic, propionic, butyric and valeric acids. There were no significant differences (p<0.05) in total gas production, composition of gas (methane and hydrogen) at all times of sampling, as well as in NH3-N production. Bacterial quantification using RT-PCR showed that the population number was not affected by transgenic corn. From this study it is concluded that transgenic corn (Mon810 and Event 176) had no adverse effects on rumen fermentation and digestibility compared to isogenic corn. However, regular monitoring of these transgenic feeds is needed by present day researchers to enable consumers with the option to select their preferred food source for animal or human consumption.

• 한국유기농업학회지
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• 제26권4호
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• pp.775-787
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• 2018

본 연구는 반추동물의 메탄 감소에 효과가 있는 합성첨가제를 대체 할 수 있는 천연첨가제를 찾고자 연구를 실시하였다. 첨가제인 MRA-1(파), MRA-2 (SLS), MRA-3 (SDS), MRA-4(파+탄닌산)를 만들어 발효시간대별(3, 6, 9, 12, 24 및 48 h) in vitro 시험을 실시하였다. pH는 발효시간동안 적정범위였으며, 건물소화율은 모든 발효시간대별 모든 처리구에서 대조구와 유의적(P>0.05)인 차이를 보이지 않았고, 미생물성장량에도 부정적인 영향을 미치지 않았다. 메탄 발생량은 발효 24시간대 MRA-1구를 제외한 나머지 처리구에서 대조구에 비해 유의적(P<0.05)으로 낮게 측정되었으며, 특히 이산화탄소 발생량은 MRA-4 첨가구에서 발효 9, 24, 48시간대에서 유의적(P<0.05)으로 낮았다. 메탄생성량 감소에 영향이 있는 프로피온산 함량은 발효 24시간대 MRA-4구에서 대조구에 비해 유의적(P<0.05)으로 높았다. Ciliate-associated methanogens은 발효 24시간대 MRA-1, MRA-3 및 MRA-4구에서 대조구에 비해 유의적(P<0.05)으로 낮게 측정되었다. 따라서 파와 탄닌산은 반추위 내 메탄 저감 효과가 있는 합성첨가제를 대체 할 수 있어 반추동물용 천연 메탄 저감제로서의 활용 가능성이 있을 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권1호
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• pp.86-91
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• 2012

An in vitro rumen batch culture study was completed to compare effects of common grasses, leguminous shrubs and non-leguminous shrubs used for livestock grazing in Australia and Ghana on $CH_4$ production and fermentation characteristics. Grass species included Andropodon gayanus, Brachiaria ruziziensis and Pennisetum purpureum. Leguminous shrub species included Cajanus cajan, Cratylia argentea, Gliricidia sepium, Leucaena leucocephala and Stylosanthes guianensis and non-leguminous shrub species included Annona senegalensis, Moringa oleifera, Securinega virosa and Vitellaria paradoxa. Leaves were harvested, dried at $55^{\circ}C$ and ground through a 1 mm screen. Serum bottles containing 500 mg of forage, modified McDougall's buffer and rumen fluid were incubated under anaerobic conditions at $39^{\circ}C$ for 24 h. Samples of each forage type were removed after 0, 2, 6, 12 and 24 h of incubation for determination of cumulative gas production. Methane production, ammonia concentration and proportions of VFA were measured at 24 h. Concentration of aNDF (g/kg DM) ranged from 671 to 713 (grasses), 377 to 590 (leguminous shrubs) and 288 to 517 (non-leguminous shrubs). After 24 h of in vitro incubation, cumulative gas, $CH_4$ production, ammonia concentration, proportion of propionate in VFA and IVDMD differed (p<0.05) within each forage type. B. ruziziensis and G. sepium produced the highest cumulative gas, IVDMD, total VFA, proportion of propionate in VFA and the lowest A:P ratios within their forage types. Consequently, these two species produced moderate $CH_4$ emissions without compromising digestion. Grazing of these two species may be a strategy to reduce $CH_4$ emissions however further assessment in in vivo trials and at different stages of maturity is recommended.

• Asian-Australasian Journal of Animal Sciences
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• 제10권4호
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• pp.357-363
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• 1997

The effect of replacement of cotton seed meal (CSM), with various levels and sources of non-protein nitrogenous (NPN), substances on in vitro ruminal fermentation were studied. Cotton seed meal, in control ration provided nitrogen equivalent to 12.5 percent crude protein while in experimental ration was replaced at 30, 50 & 70 percent levels with urea, diammonium phosphate (DAP) and biuret, respectively. The results of incubation upto 48 hours indicated an improvement in digestibility by replacement of CSM with urea and biuret upto 50 percent protein equivalent, but not with DAP. Bacterial count from cultures containing 50% nitrogen from biuret was significantly higher than DAP, urea and CSM. Various sources of nitrogen produced $NH_3-N$ in increasing order of CSM, biuret, DAP and urea. Increasing levels of NPN resulted in progressive increase in the levels of $NH_3-N$. The levels of various NPN sources had no effect on pH. However, the pH values determined for urea and CSM were higher than biuret and DAP.

• Asian-Australasian Journal of Animal Sciences
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• 제26권3호
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• pp.358-365
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• 2013

A series of in vitro studies were carried out to determine i) the effects of enzyme and formaldehyde treatment on the degradation characteristics of carbohydrate and protein sources and on the synchronicity of these processes, and ii) the effects of synchronizing carbohydrate and protein supply on rumen fermentation and microbial protein synthesis (MPS) in in vitro experiments. Untreated corn (C) and enzyme-treated corn (EC) were combined with soy bean meal with (ES) and without (S) enzyme treatment or formaldehyde treatment (FS). Six experimental feeds (CS, CES, CFS, ECS, ECES and ECFS) with different synchrony indices were prepared. Highly synchronous diets had the greatest dry matter (DM) digestibility when untreated corn was used. However, the degree of synchronicity did not influence DM digestibility when EC was mixed with various soybean meals. At time points of 12 h and 24 h of incubation, EC-containing diets showed lower ammonia-N concentrations than those of C-containing diets, irrespective of the degree of synchronicity, indicating that more efficient utilization of ammonia-N for MPS was achieved by ruminal microorganisms when EC was offered as a carbohydrate source. Within C-containing treatments, the purine base concentration increased as the diets were more synchronized. This effect was not observed when EC was offered. There were significant effects on VFA concentration of both C and S treatments and their interactions. Similar to purine concentrations, total VFA production and individual VFA concentration in the groups containing EC as an energy source was higher than those of other groups (CS, CES and CFS). The results of the present study suggested that the availability of energy or the protein source are the most limiting factors for rumen fermentation and MPS, rather than the degree of synchronicity.

• Asian-Australasian Journal of Animal Sciences
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• 제27권3호
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• pp.330-341
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• 2014

Andropogon gayanus is an important grass due to its high biomass production, drought tolerance and favorable growth on low fertility acidic soils. Currently, there is little research on the impact of growth stage on the nutritional quality or the degree of $CH_4$ production that may arise from this forage during ruminal fermentation. The objectives of this study were to determine the effects of regrowth stage of A. gayanus on its chemical composition, in vitro production of gas and CH4, as well as in vitro dry matter (DM) digestibility when grown under tropical Brazilian conditions and conserved as hay or as silage. The nutritional value of A. gayanus grass declined with increasing maturity; however digestible DM yield linearly increased. After 112 d of regrowth, A. gayanus produced higher quality silage (higher lactate and lower pH and butyrate content) and higher DM yield. However, the low levels of crude protein at this time would make protein supplementation a necessity for proper rumen fermentation. No differences in $CH_4$ kinetic parameters were found with advancing maturity or preservation method (hay or silage).

• Animal Bioscience
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• 제36권8호
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• pp.1285-1292
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• 2023

Objective: The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods: Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results: CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p<0.05) and was maintained at 37℃ to 45℃, and 0 to 250 mM NaCl. The activity of PanLys.1 significantly decreased (p<0.05) after Mn²⁺ or Zn²⁺ treatment. The relative abundance of P. anaerobius did not decrease after administration PanLys.1 under in vitro rumen conditions. Conclusion: The application of PanLys.1 to modulate P. anaerobius in the rumen might not be feasible because its lytic activity was not observed in in vitro rumen system.

• 동물자원연구
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• 제30권3호
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• pp.111-120
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• 2019

생물학적 평가 방법 중 in situ와 in vitro 평가 방법은 비용 및 시간을 절약할 수 있으며 동물 복지 측면에서 제약이 적어 주목되고 있는 방법이다. In vitro 실험에서 사용되는 사료 접종법으로 배양병에 직접 사료를 담는 dispersion 방법은 실제 반추위 발효 상황에 근접한 조건에서 실험할 수 있는 장점이 있지만, 시간과 노동력이 많이 소모된다. 반대로 사료 주머니를 이용한 방법은 실험 수행의 편이성이 있지만 첨가제 실험 수행의 제약이 있으며, 미생물 활성과 섬유소 소화율이 저하되는 단점이 있다. 본 실험은 dispersion 방법과 사료 주머니 방법을 조합하여 보다 편리한 방법으로 dispersion 에 가까운 높은 재현성의 발효 성상과 소화율 평가 가능할 것이라고 생각하여 in vitro 발효 성상과 미생물 조성을 비교하였다. Dispersion 방법과 nylon bag 을 조합한 실험에서 건물 소화율은 세척 과정에서 미생물 및 사료 입자의 유실로 과대평가될 수 있지만 이론적 최대 가스 발생량 V_max, 가스 발생량에 대한 분해 속도 상수 K_g 에서 유의적 차이가 나타나지 않으며, IVNDFD, pH, NH₃-N, total VFA 발생량 그리고 VFA 조성에서도 유의적 차이가 나지 않았다. Real-time PCR 을 이용한 미생물 조성 분석에서는 general bacteria의 절대량에서 유의적 차이가 나지 않았다. 상기의 결과를 종합해 볼 때, in vitro 실험에서 사료 주머니의 사용은 충분히 적용가능하며 일반적인 방법인 dispersion과 비교해 볼 때, 영양소 이용에 있어 반추미생물의 사료 접촉을 방해하지 않을 것으로 생각된다. 하지만 in vitro bach culture 방법은 배양조건에서 제약이 있어, 향후 연속 배양 방법과 in situ의 실험을 통해 본 실험의 실험결과에 대한 추가적인 규명이 필요할 것으로 사료된다.