• 한국식물학회:학술대회논문집
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• 한국식물학회 1996년도 제10회 식물생명공학심포지움 고등식물 발생생물학의 최근 진보
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• pp.61-74
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• 1996

Human and monogastric animals can not synthesize 10 out of the 20 amino asids and therefor need to obtain these from their diet. The plant seed is a major source of dietary protein. It is particular important in their study to increase nutritional quality of the seed storage proteins. The low contents of lysine, asparagine and threonenein various cereal seeds and of cystein and methionine. In legume seeds is due to the low proportions of these amino acids in the major storage proteins, we have tried to apply the three strategies; (1) mutagenesis and selection of specific amino acid analogue resistance, (2) cloning and expression study of lysine biosynthesis related gene, (3) transfomation of lysine rich soybean glycinin gene. The 5-methyltryptophan (5MT) resistant cell lines, SAR1, SAR2 and SAR3 were selected from anther derived callus of rice (Oryza sativa L. "Sasanishiki"). Among these selected cell lines, two (SAR1 and SAR3) were able to grow stably at 200 mg/L of 5MT. Analysis of the freed amino acids in callus shows that 5MT resistant cells (SAR3) accumulated free tryptophan at least up to 50 times higher than those that of the higher than of SAS. These results indicated that the 5MT resistant cell lines are useful in studies of amino acid biosynthesis. Tr75, a rice (Oryza sativa L., var. Sasanishiki) mutant resistant to 5MT was segregated from the progenies of its initial mutant line, TR1. The 5MT resistant of TR75 was inherited in the M8 generations as a single dominant nuclear gene. The content of free amino acids in the TR75 homozygous seeds increased approximately 1.5 to 2.0 fold compared to wild-type seeds. Especially, the contents of tryptophan, phenylalanine and aspartic acid were 5.0, 5.3 and 2.7 times higher than those of wild-type seeds, respectively. The content of lysine is significantly low in rice. The lysine is synthesized by a complex pathway that is predominantly regulated by feedback inhibition of several enzymes including asparginase, aspatate kinase, dihydrodipicolinat synthase, etc. For understanding the regulation mechanism of lysine synthesis in rice, we try to clone the lysine biosynthetic metabolism related gene, DHPS and asparaginase, from rice. We have isolated a rice DHPS genomic clone which contains an ORF of 1044 nucleotides (347 amino acids, Mr. 38, 381 daltons), an intron of 587 nucleotides and 5'and 3'-flanking regions by screening of rice genomic DNA library. Deduced amino acid sequence of mature peptide domain of GDHPS clone is highly conserved in monocot and dicot plants whereas that of transit peptide domain is extremely different depending on plant specie. Southern blot analysis indicated that GDHPS is located two copy gene in rice genome. The transcripts of a rice GDHPS were expressed in leaves and roots but not detected in callus tissues. The transcription level of GDHPS is much higher in leaves indicating enormous chloroplast development than roots. Genomic DNA clones for asparaginase genes were screened from the rice genomic library by using plaque hybridization technique. Twelve different genomic clones were isolated from first and second screening, and 8 of 12 clones were analyzed by restriction patterns and identified by Southern Blotting, Restriction enzyme digestion patterns and Southern blot analysis of 8 clones show the different pattern for asparaginase gene. Genomic Southern blot analysis from rice were done. It is estimated that rice has at least 2-3 copy of asparaginase gene. One of 8 positive clones was subcloned into the pBluescript SK(+) vector, and was constructed the physical map. For transformation of lysine rich storage protein into tobacco, soybean glycinin genes are transformed into tobacco. To examine whether glycinin could be stably accumulated in endosperm tissue, the glycinin cDNA was transcriptionally fused to an endosperm-specific promotor of the rice storage protein glutelin gene and then introduced into tobacco genomic via Agrobacterium-mediated transformation. Consequently the glycinin gene was expressed in a seed-and developmentally-specific manner in transgenic tobacco seeds. Glycinin were targeted to vacuole-derived protein bodies in the endosperm tissue and highly accumulated in the matrix region of many transgenic plant (1-4% of total seed proteins). Synthesized glycinin was processed into mature form, and assembled into a hexamer in a similar manner as the glycinin in soybean seed. Modified glycinin, in which 4 contiguous methionine residues were inserted at the variable regions corresponding to the C - teminal regions of the acidic and basic polypeptides, were also found to be accumulated similarly as in the normal glycinin. There was no apparent difference in the expression level, processing and targeting to protein bodies, or accumulation level between normal and modified glycinin. glycinin.

• 한국식품영양과학회지
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• 제38권1호
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• pp.76-82
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• 2009

본 연구에서는 발효과정 중 누룩의 일반성분 및 효소 활성변화를 측정하고, 단백질의 분해정도와 밀 단백질에 민감한 환자의 혈청과 반응성을 관찰하였다. 누룩의 수분함량은 발효 1일 24.20%였으며 발효가 진행됨에 따라 미생물의 이용과 증발로 인해 감소되어 발효 15일에는 13.60%를 나타내었다. 누룩 발효 중 총당과 환원당의 함량은 발효시간이 경과함에 따라 증가하는 경향을 나타내었다. 누룩의 발효 초기 조단백질 함량은 1.58%였으나 발효가 진행됨에 따라 다소증가하여 발효 15일에는 2.05%로 나타났다. 또한 누룩의 조지방 함량은 발효 초기 1.32%였으며 발효 15일에는 2.24%로 다소 증가함을 나타내었다. ${\alpha}$-Amylase 활성은 발효 초기 1,416.67 U/g에서 발효 15일에는 2,833.00 U/g으로 급격히 증가하였으며, glucoamylase 활성은 발효 초기 497.90 U/g에서 발효 10일 경과 후 705.40 U/g으로 최대 활성을 보였으나 그 후 완만한 감소를 나타내었다. 누룩의 산성 protease 활성은 발효 초기에는 검출되지 않았으나 발효 10일에 75.00 U/g으로 급격히 증가하기 시작하여 발효 15일에는 327.00 U/g에 도달하였으며 중성 protease 활성도 발효 10일에 90.00 U/g으로 급격히 증가하기 시작하여 15일에는 354.00 U/g로 검출되었으며 이후에도 계속 증가하였다. 반면에 알칼리성 protease 활성은 검출되지 않았다. 발효가 진행됨에 따라 누룩 중 고분자 단백질이 각종 미생물에 의해 분해되어 저분자의 펩타이드 밴드가 생성되었으며 밀 민감성 환자의 혈청과 반응하는 밴드도 점차 감소하여 발효 15일에는 거의 검출되지 않았으므로 누룩 중 밀 단백질의 항원성이 감소하였음을 확인할 수 있었다. 따라서 본 연구를 통해 누룩은 영양학적 측면뿐만 아니라 효소학적 측면의 장점까지 겸비한 효율적인 한국 고유의 발효 식품용 스타터(starter)로써 전통주 제조에 필수적이며, 누룩의 고분자 단백질이 발효에 의해 저분자 펩타이드로 분해됨을 확인함으로써 밀 민감성 환자들을 위한 다양한 가공식품용 소재로 활용이 기대된다.

• 한국가금학회지
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• 제31권4호
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• pp.283-291
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• 2004

본 연구에서는 생물학적 탄산가스 고정화에 사용되는 Euglena를 사료자원으로 이용하고, Euglena의 DHA를 강화시켜 산란계의 사료에 첨가하여 그 이용성과 난황내 지방산 조성에 미치는 영향에 대해 알아보고자 실시하였다. 시험 1에서는 32주령의 산란계(ISA-Brown) 280수를 7처리로 나누어 처리당 4반복으로 반복당 10수씩 배치하였다. 처리구는 에너지함량과 조단백질함량이 2,750 kcal/kg과$17\%$인 대조구 사료에 EG(Euglena gracilis Z.)를 0.25, 0.50, $1.0\%$ 첨가한구와 EGBD(Euglena gracilis B bleached and DHA enriched)를 0.5, 1.0, $2.0\%$ 첨가한 구의 7처리구로 하였다. 시험 2에서는 84주령의 산란계 300수를 5처리로 나누어 처리당 5반복으로 반복당 12수씩 배치하였다. 처리구는 대조구사료에 EGBD를 $0.5\%$ 첨가한 구와 EGD(Euglena gracilis Z. DHA enriched)를 0.5, 1.0, $2.0\%$ 첨가한 구의 5처리구로 하였다. 두시험 모두 사양시험은 4주간 실시하였고, 시험기간동안 물과 사료는 자유로이 섭취케 하고 정상적인 점등관리를 실시하였다. 시험 1의 결과를 보면,c일계산란율과 산란지수 모두 유의한 차이를 보이지 않았고, 난중은 시험사료 급여결과 $1.0\%$의 EG 첨가구가 가장 무거웠으며, 대조구가 가장 가벼웠다. 난황색은 모든 처리구가 대조구보다 유의하게 높았으며, EG를 첨가한 구들의 난황색이 높은 경향을 나타냈다. 지방산 조성을 보면 $2.0\%$ EGBD 첨가구의 DHA와 EPA의 수준이 유의적으로 가장 높았으며, arachidonic acid의 수준은 가장 낮았다. 시험 2에서도 일계 산란율과 산란지수 모두 유의한 차이를 보이지 않았고, 난중은 $0.5\%$의 EGD를 첨가한 구가 유의적으로 가장 높았으며, 대조구를 포함한 다른 처리구들에 비해 EGD를 첨가한 구들이 높은 경향이 있었다. 지방산 조성의 결과를 보면 $2.0\%$ EGD 첨가구의 myristic acid, myristoleic acid와 pentadecanoic acid가 모두 유의적으로 가장 높았다. DHA는 $0.5\%$%의 EGBD 첨가구가 유의적으로 가장 높았으며, EGD를 첨가한 구들이 대조구에 비해 유의하게 높았다. 결론적으로, Euglena의 첨가는 산란계의 생산성에는 크게 영향을 미치지 못했지만, EG의 첨가가 난황색의 개선에 효과가 있었고, EG, EGD 그리고 ECBD 모두가 난중을 높여주는 경향이 있었다. 또한, Euglena의 DHA를 강화시킨, EGD나 특히 EGBD의 첨가는 난황내의 DHA를 비롯한 $\omega%-3 계열의 지방산의 수준을 높이는데 효과적이라고 사료되어진다.

• Asian-Australasian Journal of Animal Sciences
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• 제22권5호
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• pp.651-658
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• 2009

Four multiparous lactating Holstein cows fitted with rumen cannulae were fed diets varying in the amount and source of rumen-degradable carbohydrates (starch vs. sucrose) to examine their effects on rumen fermentation, nitrogen metabolism and lactation performance. A $4{\times}4$ Latin square with four diets and four periods of 28 days each was employed. Corn starch and sucrose were added to diets and corn starch was replaced with sucrose at 0 (0 S), 2.5 (2.5 S), 5.0 (5.0 S) 7.5% (7.5 S) of diet dry matter in a total mixed ration (TMR) containing 60% concentrate and 40% forage (DM basis). Replacing corn starch with sucrose did not affect (p>0.05) ruminal pH which averaged 6.41, but the ruminal pH for 7.5 S decreased more rapidly at 2 h after morning feeding compared with other treatments. Sucrose reduced ($p{\leq}0.05$) ruminal $NH_3-N$ concentration (13.90 vs. 17.09 mg/dl) but did not affect peptide-N concentration. There was no dietary effect on total volatile fatty acids (110.53 mmol/L) or the acetate to propionate ratio (2.72). No differences (p>0.05) in molar proportion of most of the individual VFA were found among diets, except for the molar proportion of butyrate that was increased ($p{\leq}0.05$) with the inclusion of sucrose. Total branched chain volatile fatty acids tended to increase ($p{\geq}0.051$) for the control treatment (0 S) compared with the 7.5 S treatment. Dry matter intake, body weight changes and digestibility of DM, OM, CP, NDF and ADF were not affected by treatments. Sucrose inclusion in the total mixed ration did not affect milk yield, but increased milk fat and total solid percentage ($p{\leq}0.05$). Sucrose tended ($p{\geq}0.063$) to increase milk protein percentage (3.28 vs. 3.05) and reduced ($p{\leq}0.05$) milk urea nitrogen concentration (12.75 vs. 15.48 mg/dl), suggesting a more efficient utilization of the rapidly available nitrogen components in the diet and hence improving nitrogen metabolism in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• 제21권4호
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• pp.547-554
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• 2008

In performance, digestibility and slaughter trials, a total of forty five male weaned lambs were used to examine the effects of monensin and live yeast supplementations on growth performance, nutrient digestibility, carcass characteristics and ruminal fermentation parameters when the lambs were fed steam-flaked corn-based diets. Animals were allotted to one of three treatment diets in a completely randomized design. The three treatment diets were: (1) basal diet (CON) with steam-flaked corn as a sole grain source, (2) basal diet supplemented with monensin (MO), and (3) basal diet supplemented with live yeast (LY). Total average daily intake (ADI) was unaffected by MO and LY supplementations. LY supplementation increased (p<0.05) average daily gain (ADG) by 13.1% compared with the CON diet. Both MO and LY supplementations resulted in a significant improvement (p<0.05) of feed efficiency over the CON diet (4.47, 4.68 vs. 5.05). Hemicellulose digestibility was higher (p<0.05) for lambs in the LY supplementation group (62.4%) as compared with the CON group (55.7%), but no differences were observed in digestibilities of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF). All carcass traits were not influenced by dietary supplementations. Ruminal pH in lambs fed the LY supplemental diet was more stable than that with the CON diet (6.57 vs. 6.17). Neither MO nor LY supplementation influenced the concentration of ruminal ammonia-N and total volatile fatty acid (VFA), and molar percentages of individual VFA. Plasma urea-N concentration was decreased (p<0.05) by MO and LY supplementations, while plasma ${\beta}$-hydroxybutyrate (BHBA), glucose and other blood parameters were unaffected. In conclusion, while both MO and LY supplementations had a positive impact on feed efficiency and LY supplementation stabilized ruminal pH and improved fiber utilization, none of the supplements had the capacity to significantly enhance the carcass characteristics.

• Asian-Australasian Journal of Animal Sciences
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• 제31권6호
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• pp.888-898
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• 2018

Objective: The study was conducted to investigate the effects of replacing soybean meal (SBM) with different levels of detoxified Jatropha curcas kernel meal (DJM) in growing pig diets on growth performance, nutrients digestibility and meat edibility. Methods: A total of 144 pigs with initial body weight of $20.47{\pm}1.44kg$, were randomly allocated to 6 dietary treatments with 6 replications per treatment and 4 pigs per replication for a period of 79 days. Six diets (DJM0, DJM15, DJM30, DJM45, DJM60, and DJM75) were formulated using DJM to replace 0%, 15%, 30%, 45%, 60%, and 75% of SBM. From d 37 to 42, feces and urine were total collected from six barrows in each treatment. At day 79, thirty-six pigs were slaughtered for sampling. The feed intake and weight gain were recorded, while the intestinal morphology, digestive enzyme activities, nutrient digestibility and the content of residual phorbol esters in muscles were determined. Results: The results showed that increasing the replacement of SBM with DJM decreased the parameters including body weight, average daily gain, average daily feed intake, gain-to-feed ratio, weight and villus heights of duodenum, villus height and villus height/crypt depth of jejunum, digestive enzymes (protease, amylase, lipase, and trypsin) activities, and nutrients digestibility (nitrogen deposition, digestibility of nitrogen, energy digestibility, and total nitrogen utilization) (linear, p<0.05; quadratic, p<0.05) and there was no significant difference among DJM0, DJM15, and DJM30 in all measured indices. The highest diarrhea morbidity was observed in DJM75 (p<0.05). Phorbol esters were not detected in pig muscle tissues. Conclusion: The DJM was a good protein source for pigs, and could be used to replace SBM up to 30% (diet phorbol esters concentration at 5.5 mg/kg) in growing pig diets with no detrimental impacts on growth performance, nutrient utilization, and meat edibility.

• Asian-Australasian Journal of Animal Sciences
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• 제24권7호
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• pp.957-964
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• 2011

The present study was conducted to evaluate the effects of fermented soybean meal (FSBM) on the level of cortisol hormone and immune-related serum proteins in weaned calves after experimentally induced lipopolysaccharide (LPS) challenge. Holstein neonatal calves (n = 21; 8 males and 13 females, BW = $42.2{\pm}6.15$ kg) were randomly allocated to one of two dietary treatments: SBM (control calf starter having soybean meal (SBM) as a main protein source) and FSBM (substitute SBM in control diet with FSBM) groups. All calves were fed milk replacer using an automatic milk-feeder according to step-down milking method and weaned at 7 weeks old. Experimental diets were given to calves ad libitum throughout the experimental period. For LPS challenge, all calves except negative control animals given phosphate buffered saline (PBS), were injected subcutaneously with Salmonella typhimurium LPS on day 7 (D7) after weaning (D0). No significant difference in growth performance and milk intake was observed between SBM and FSBM calves. Feeding FSBM diet resulted in significantly (p<0.05) higher LPS-specific IgG at D12 and D19 and LPS-specific IgA at D19 in peripheral blood. Calves fed with FSBM diet also had significantly (p<0.05) higher concentration of serum haptoglobin (Hp) at D8. Overall concentration of cortisol in FSBM group was considerably lower than that of SBM group. Results from current study indicate that FSBM may provide beneficial effects in alleviating weaning stress and enhance immune status of weaned calves.

• Asian-Australasian Journal of Animal Sciences
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• 제26권1호
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• pp.59-64
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• 2013

Eight male cattle of the Local Yellow breed with an average live weight of 121 kg and an average age of 18 months were used to evaluate the effects of different levels of sun-dried cassava (Manihot esculenta) foliage supplementation on mineral metabolism in growing cattle fed rice straw and para grass as basal diet. Rice straw ad libitum and para grass (Brachiaria mutica) at 1% DM of BW comprised the basal diet. The study was arranged as a $4{\times}4$ double Latin square design, with cassava foliage contributing 0, 0.8, 1.6 or 2.4 g CP/kg BW. The cassava foliage intake was lower than the planned levels. DM consumption was significantly affected by cassava foliage supplementation, with the largest intake observed at the two highest levels of cassava foliage supplementation. Rice straw intake showed the opposite pattern, with lower intake at higher cassava foliage supplementation. No refusals occurred for para grass in any of the treatments. Ca, P, Mg, K, S and Mn intake increased significantly with increasing intake of cassava foliage, but Na intake was not affected by treatment. Faecal excretion of Ca, Mg, S and Mn increased significantly with increasing cassava foliage intake. There were no differences between P, K and Na excretion in faeces. There was a significant diet effect on Mg, S and Mn digestibility. Mg and Mn digestibility increased with increasing cassava foliage supplementation, while S digestibility decreased. Ca, P, K and Na digestibility was not affected by diet. There was a significant effect of treatment on P retention, with the highest value observed for supplementation with 1.6 g CP/kg BW cassava foliage. Ca and Mg showed similar trends, with the highest retention again for supplementation with 1.6 g CP/kg BW cassava foliage. There were weak but significant positive correlations between nitrogen retention and the macro minerals Ca, P and Mg. Furthermore, retention of all these minerals was positively correlated. Mineral losses in urine were not affected by dietary treatment with the exception of P excretion, which was affected by treatment. In conclusion, cassava foliage is a good Ca source which compensates for the low Ca content in rice straw and para grass, but P deficiency appears to be exaggerated in cattle with higher cassava intake. The results suggest that under these conditions growing cattle on a high cassava intake would benefit from P and S supplementation.

• 한국식품영양과학회지
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• 제39권11호
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• pp.1627-1633
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• 2010

본 연구는 더덕껍질의 기능성식품 및 화장품 소재로서의 이용가능성을 조사하기 위해서 더덕껍질의 영양성분 분석을 통한 식품영양학적 접근, 생리활성 기능을 기대할 수 있는 관련 물질 함량을 분석하였다. 식품영양학적 접근에서의 더덕껍질의 일반성분은 건량기준으로 탄수화물 24.74%, 조단백질 2.73%, 조지방 2.96% 및 조회분 4.84%이었고 더덕껍질 100 g의 함유 열량은 266.00 kcal로 분석되었으며, 총 식이섬유소 함량은 건량기준으로 64.73%로 나타났다. 또한, 필수아미노산과 비필수아미노산 함량은 각각 633.40과 870.72 mg/100 g wet weight basis이었고, 무기질 중 칼륨의 함유량이 가장 높았고 그 다음이 칼슘, 마그네슘, 인순으로 나타나 알칼리성 재료임을 알 수 있었으며, 지방산 함량의 경우 불포화지방산의 함량이 높은 것으로 나타났다. 유리당의 경우 fructose의 함량이 전체 유리당의 약 72%를 차지하고 있는 것으로 나타났다. 더덕껍질 물추출물의 총 페놀화합물 함량은 $24.65\;{\mu}g/g$, 플라보노이드 함량은 $6.19\;{\mu}g/g$으로 분석되었으며, DPPH 소거 활성을 농도별로 측정하여 비교 한 결과 $500\;{\mu}g/mL$ 농도에서 79.30%로 높은 활성을 보였다. 또한, 환원력의 경우에는 더덕껍질 추출물이 25 mg/mL의 농도에서 3.5로 높은 환원력을 보였다. 이상의 결과로부터 더덕껍질 물추출물은 항산화능이 있음을 확인할 수 있었다. 따라서 더덕껍질을 식품 내 첨가물로서, 혹은 다른 약용으로의 활용가능성을 제시하고 있다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권8호
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• pp.1124-1131
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• 2012

Four Thai - rumen fistulated male swamp buffaloes (Bubalus bubalis), about four years old with $400{\pm}20kg$ liveweight, were randomly assigned according to a $4{\times}4$ Latin square design to receive dietary treatments. The treatments were: ground corn cob (GCC) replacement for cassava chip (CC) in concentrate at 0% (T1); GCC replacement at 33% (T2); GCC replacement at 67% (T3); and GCC replacement at 100% (T4), respectively. During the experiment, concentrate was offered at 0.5% BW while 5% urea-treated rice straw was given at ad libitum. The result revealed that there was no effect of GCC replacement on DMI among treatments. In addition, digestibilities of DM, OM and CP were not different while aNDF linearly increased with an increasing level of GCC replacement. However, GCC replacement did not affect rumen fermentation such as ruminal pH, $NH_3$-N and VFA concentration; except C3 proportion which was the highest at 33% replacement while the lowest was at 100% replacement. All replacements of GCC resulted in similar protozoal and bacterial populations and microbial protein synthesis (MPS). Purine derivatives (PD) concentration in urine and PD to creatinine (PDC) index were varied with time of urination and among treatments at 0 to 8 and 8 to 16 h post feeding and higher values were shown among the GCC replacement groups. However at 16 to 24 h-post feeding, it was untraceable. In addition, creatinine concentration was similar among all treatments at every sampling time. Based on the above results, GCC can be used as an energy source for swamp buffalo fed with rice straw. Spot sampling of urine can be used for purine derivatives determination.