This study was conducted to investigate the relationship between changes of rumen microflora and bloat in Jersey cow. Jersey cows (control age: 42 months, control weight: 558kg; treatment age: 29 months, treatment weight 507kg) were fed on the basis of dairy feeding management at dairy science division in National Institute of Animal Science. The change of microbial population in rumen was analyzed by using next generation sequencing (NGS) technologies due to metabolic disease. The diversity of Ruminococcus bromii, Bifidobacterium pseudolongum, Bifidobacterium merycicum and Butyrivibrio fibrisolvens known as major starch fermenting bacteria was increased more than 36-fold in bloated Jersey, while cellulolytic bacteria community such as Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens was increased more than 12-fold in non-bloated Jersey. The proportion of bacteroidetes and firmicutes was 33.4% and 39.6% in non-bloated Jersey’s rumen, while bacteroidetes and firmicutes were 24.9% and 55.1% in bloated Jersey’s. In conclusion, the change of rumen microbial community, in particular the increase in starch fermenting bacteria, might have an effect to occur the bloat in Jersey cow.

The gene encoding an esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est2R) was 2,120 bp in length, encoding a protein of 516 amino acid residues with a calculated molecular weight of 57,286 Da. The molecular weight of the enzyme was estimated to be 57,000 Da by SDS-PAGE. Est2R shared 35.6% amino acid identity with esterase (CAH19079) of uncultured prokaryote. The Est2R was most active at 20-40°C, and showed optimum at 30°C and pH 8.0. The most activity of Est2R for the different chain length of p-nitrophenyl ester group as substrate was p-nitrophenyl acetate. Moreover, the enzyme was found to be most active without organic solvent, followed by 98% active with ethanol, and the enzyme activity was highly affected by the acetonitrile. The enzyme was significantly inhibited by Zn2+ but stimulated by Ca2+. So, novel esterase gene est2R is likely to obtain from cow rumen metagenome and supposed to use for industrial purpose.

A carboxymethyl cellulase gene, cel5B, was cloned, sequenced, and expressed in Escherichia coli. pRCS20 in E. coli was identified from metagenomic cosmid library of cow rumen for cellulase activity on a carboxymethyl cellulose agar plates. Cosmid clone (RCS20) was partially digested with Sau3AI, ligated into BamHI site of pBluescript II SK+ vector, and transformed into E. coli DH5α. The insert DNA of 1.3 kb was obtained, designated cel5B, which has the activity of hydrolyzation of CMC. The cel5B gene had an open reading frame (ORF) of 1,059 bp encoding 352 amino acids with a signal peptide of 48 amino acids and the conserved region, VIYEIYNEPL, belongs to the glycosyl hydrolase family 5. The molecular mass of Cel5B protein expressed from E. coli DH5α exhibited to be about 34 kDa by CMC-SDS-PAGE. The optimal pH was 8.0, and the optimal temperature was about 50℃ for its enzymatic activity.

우리나라 고유 식품인 소양의 영양적 가치와 최적조리 조건을 제시하기 위해서, 소양 조직의 일반 영양성분과 조리방법(가열처리, 압력처리, 양과 물의 첨가비)에 따른 질소성분과 미네랄성분의 용출, 핵산관련 맛성분의 용출 및 관능검사 결과를 요약하면 다음과 같다. 1. 소양의 제 1위와 제 2위의 영양성분은 거의 차이가 없었으며 수분 약 83%, 회분 0.4~0.5%, 지방 약 3%, 단백질 약 13%, 칼슘 50~56mg%, α-amino N 75~76mg%, 인 75~76mg%였다. 소양의 상피세포층의 중량은 제 1위에서 35%, 제 2위에서 29%를 차지하였으며, 회분과 칼슘을 제외한 모든 영양성분은 상피세포층과 근육층에 균일하게 분포되어 있었으나 칼슘은 총함량의 80% 이상이 상피세포층에 함유되어 있었다. 2. 소양을 2, 4, 8, 12, 24시간 가열한 후 질소성분(총질소와 α-amino N)과 미네랄성분(칼슘과인)은 각각 8시간과 4시간 가열에 의해 plateau에 달하였으며, 맛성분인 5'-IMP는 8시간 가열에 의해 plateau에 달하였다. 관능검사 결과는 8-12시간 가열시 좋은 성적을 얻었다. 3. 소양을 압력솥에서 30분, 1, 2, 3, 4시간 가열조리한 후 질소성분과 미네랄성분은 각각 2시간과 1시간 가열에 의해 plateau에 달하였으며 5'-IMP는 2시간의 압력가열로 최대값을 나타냈다. 관능검사 결과는 1시간 이상 가열시 좋은 성적을 나타내었다. 4. 소양의 중량별 용출액 중의 영양성분을 보면, 양에 대한 물의 첨가량이 10배 이상일 때 용출량은 증가하고, 특히 칼슘은 10배 이하에서는 20배에서의 용출량의 50%로 감소되었다. 맛성분의 용출은 20배에서 가장 효과적이었으며, 관능검사 결과에는 차이가 없었다. 이상의 실험 성적에서 소양은 단백질과 칼슘의 급원 식품으로서 조리시 상피세포층을 제거하였을 때 영양적 손실이 크며, 특히 칼슘의 손실이 컸다. 영양성분의 충분한 용출과 맛을 위해서는 상압가열 조리에 있어서는 8시간의, 압력조리에 있어서는 1~2%시간의 조리시간, 양에 대한 물의 첨가량은 10배 이상이 필요하였다. 이것은 소양의 조리시 충분량의 물을 넣어 장시간 가열하는 우리나라 재래식 조리방법이 영양적, 미각적 측면에서 매우 합리적임을 시사한 것으로 본다.

This study was carried out to investigate the effect of DDGS hydrolysate (H-DDGS) and rumen-protected lysine-choline complex (RPLC) on milk production and blood metabolites in dairy cows. Feeding trials was performed to latin-square design using the 4 mid-lactational cows for 8 weeks, and treated with T1 (H-DDGS 1.1 kg), T2 (H-DDGS 0.73 kg + RPLC 0.15 kg), T3 (H-DDGS 0.37 kg + RPLC 0.30 kg) and T4 (H-DDGS 0.37 kg + RPLC 0.15 kg) according to the content of protein source. Dry matter intake (DMI) of TMR and average weight showed no significant difference between treatments. The milk production of T1, T2 and T4 were significantly higher than T3 treatment (p < 0.05), and milk/DMI efficiency tend to increase in the T1. Milk components showed no significant difference between treatments, however, the milk protein of T2 increased to 0.15% than T1. Also blood metabolites showed no significant difference between treatments. But T-CHO level numerically represented a lower trend in the treatments of adding to RPLC compared with T1. This result suggests that the high level (1.1 kg) of H-DDGS is expected to improve the feed utilization without the negative impact on weight gain, feed intake and milk production as the lactation stage of dairy cows proceeds, and 0.15 kg of RPLC under the same feeding conditions of H-DDGS may be useful on fat metabolism.