• Asian-Australasian Journal of Animal Sciences
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• v.21 no.9
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• pp.1367-1375
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• 2008

This study was conducted to investigate the effects of phytase in diets containing cottonseed and soybean meal (CS) on growth performance, feed utilization and digestibility of protein and phosphorus in juvenile olive flounder (initial body weight 2.5 g), Paralichthys olivaceus. Four experimental diets replacing 0%, 30%, 30% and 40% fish meal protein with CS in equal proportion were formulated to be isonitrogenous and isocaloric (designated as CS0, CS30, CS30+P, CS40+P, respectively). Phytase of 1,000 FTU/kg was supplemented in diets CS30+P and CS40+P. Three groups of fish (25 fish per group) were fed one of the experimental diets for 10 weeks. No significant differences were observed in growth performance of fish groups except for the CS40+P diet. Apparent digestibility coefficients of protein and phosphorus in fish fed phytase-containing diets were significantly higher than those of fish fed the CS0 diet. Serum cholesterol was significantly reduced in fish fed the CS-containing diets. Antioxidant activities in the diets and liver of fish were significantly increased with the increment of dietary CS. Gossypol was only detected and found in liver of the fish fed the CS-containing diets. The findings suggest that supplementation of microbial phytase could improve the apparent digestibility of protein and phosphorus in juvenile olive flounder fed the CS-containing diets.

• Journal of Microbiology and Biotechnology
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• v.11 no.3
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• pp.534-538
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• 2001

The xynA gene encoding an alikali-tolerant endo-1,4-${\beta}$-xylanase (XYN) was cloned from the alkalophilic Bacillus pumilus A-30. The nucleotide sequence of a 974-bp DNA fragment containing the xynA was determined. An ORF of 684 nucleotides that encoded a protein of 228 amino aicds was detected. Asparagine-71 of XYN from B. Pumilus A-30 showed to be highly conservative in alkaline xylanases of family G/11, upon comparing the amino acid sequences of 17 family G/11 xylanases. Site-directed mutation of N71D of the xynA gene resulted in a decrease of 12.4% in the specific acitivity and a significant decline in the enzyme activity in the alkaline pH range.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.3
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• pp.347-355
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• 2006

A study was conducted to predict the rumen microbial protein production based on urinary excretion of purine derivatives in buffaloes fed a diet of wheat straw and concentrate (40:60) at four fixed levels of feed intake. (95, 80, 60 and 40% of preliminary voluntary feed intake) following experimental protocol of IAEA (Phase I). The buffaloes were allocated according to a $4{\times}4$ latin square design. The urinary allantoin, uric acid, total PD excretion (mmol/d) in treatments L-95, L-80, L-60 and L-40 was 20.13, 16.00, 12.96 and 9.17; 1.88, 2.12, 2.11 and 2.15; 22.01, 18.12, 15.07 and 11.32, respectively and were significantly (p<0.05) different among treatments except for uric acid. The rate of PD excretion (mmol/d) was positively correlated with the digestible organic matter intake. Variations were observed in PD and creatinine concentration in spot samples collected at 6-hour interval. However, daily PD:Creatinine ratio (PDC index) appears to be a reasonably good predictor of microbial-N supply. The contribution of basal purine excretion to total excretion of purine derivatives (PD) was determined in pre-fasting period followed by a fasting period of 6 d (Phase II). Daily PD and creatinine excretion (mmol/kg $W^{0.75}$) during fasting averaged 0.117 and 0.456 respectively for buffaloes. The excretion rates of PD decreased significantly (p<0.01) during fasting compare to pre-fasting period, the urinary creatinine excretion remained almost similar. Except for creatinine, plasma concentration of target parameters significantly (p<0.01) declined during fasting. Likewise, glomerular filtration rate (GFR) and renal clearance of allantoin and uric acid also decreased. Based on the PD excretion rates during fasting and at different levels of feed intake obtained in this study, a relationship between daily urinary PD excretion (Y-mmol) and microbial purine absorption (X-mmol) was developed for buffaloes as Y = 0.74X+0.117 kg $W^{0.75}$. The microbial N supply (g/kg DOMI) remained statistically similar irrespective of dietary treatment. The results showed that excretion of urinary purine derivatives is positively correlated with the levels of feed intake in Murrah buffaloes and thus, estimation of urinary purine derivatives and PDC index could be used to determine microbial nitrogen supply when there is large variation in level of feed intake.

• Proceedings of the Microbiological Society of Korea Conference
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• 2003.05a
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• pp.9-9
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• 2003

• Applied Biological Chemistry
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• v.48 no.4
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• pp.345-351
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• 2005

To access the natural products of uncultured microorganisms, we constructed and screened the metagenomic DNA libraries by using a cosmid vector and DNA inserts isolated directly from soil. Initial screening of the libraries in Escherichia coli resulted in the isolation of several clones that produce a dark brown color when grown in LB medium. One of the positive clones, designed pYS85C, was transposon mutagenized and the DNA surrounding the transposon insertions in cosmids that no longer conferred the production of brown pigment to E. coli was sequenced. Annotation of the pYS85C sequence obtained from the transposon mutagenesis experiment indicated a single 393 amino acid open reading frame (ORF) with a molecular mass of about 44.5 kDa, predicted to be a 4-hydroxyphenylpyruvate dioxygenases (HPPDs), was responsible for the observed brown pigment. In a BLAST search against deposited sequence, the translated protein from this ORF showed moderate-level identity (>60%) to the other known HPPDs and was most conserved in the C-terminal region of the protein. These results show that genes involved in natural product synthesis can be cloned directly from soil DNA and expressed in a heterologous host, supporting the idea that this technology has the potential to provide novel natural products from the wealth of environmental microbial diversity and is a potentially important new tool for drug discovery.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.8
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• pp.1144-1151
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• 2013

In this study, protein domains with cellulase activity in goat rumen microbes were investigated using metagenomic and bioinformatic analyses. After the complete genome of goat rumen microbes was obtained using a shotgun sequencing method, 217,892,109 pair reads were filtered, including only those with 70% identity, 100-bp matches, and thresholds below $E^{-10}$ using METAIDBA. These filtered contigs were assembled and annotated using blastN against the NCBI nucleotide database. As a result, a microbial community structure with 1431 species was analyzed, among which Prevotella ruminicola 23 bacteria and Butyrivibrio proteoclasticus B316 were the dominant groups. In parallel, 201 sequences related with cellulase activities (EC.3.2.1.4) were obtained through blast searches using the enzyme.dat file provided by the NCBI database. After translating the nucleotide sequence into a protein sequence using Interproscan, 28 protein domains with cellulase activity were identified using the HMMER package with threshold E values below $10^{-5}$. Cellulase activity protein domain profiling showed that the major protein domains such as lipase GDSL, cellulase, and Glyco hydro 10 were present in bacterial species with strong cellulase activities. Furthermore, correlation plots clearly displayed the strong positive correlation between some protein domain groups, which was indicative of microbial adaption in the goat rumen based on feeding habits. This is the first metagenomic analysis of cellulase activity protein domains using bioinformatics from the goat rumen.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.1
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• pp.65-72
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• 2011

Twenty-four Holstein dairy cows were used to evaluate the single and combined effects of different levels of crude protein (CP) and monensin treatment during early lactation on blood metabolites, milk yield and digestion of dairy cows. The experiment was designed as a completely randomized block with a $3{\times}2$ factorial arrangement of treatments. The factors were three concentrations of CP supplement (19.5, 21.4, and 23.4% of dry matter) and two levels of monensin (0 and 350 mg per cow per day). The experiment consisted of three phases and each phase was 3 wk in length. Monensin did not affect milk yield, lactose, solids-non-fat (SNF), blood glucose, triglyceride and DMI, but increased blood cholesterol, blood urea nitrogen (BUN), insulin and reduced blood ${\beta}$-hydroxybutyrate (BHBA), milk fat and protein percentage. Monensin premix significantly decreased rumen ammonia, but rumen pH and microbial protein synthesis were not affected by monensin treatment. Increasing dietary CP improved milk and protein production, but did not alter the other components of milk. Digestibility of NDF, ADF, CP were improved by increasing dietary CP. Increasing dietary CP from 19.5 to 21.4% had no significant effect on ruminal ammonia, but increasing CP to 23.4% significantly increased ruminal ammonia. There was a linear relationship between level of crude protein in the diet and volume of urine excretion. Microbial protein synthesis was affected by increasing CP level; in this way maximum protein synthesis was achieved at 23.4% CP.

• Journal of Animal Science and Technology
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• v.47 no.3
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• pp.419-428
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• 2005

This study was carried out to investigate the effect of replacing dried mugwort (Artemisia montana Pampan) in concentrate a time of preparation pellet. The treatments, arranged in a 4${\times}$4 Latin square design, were four additional levels of mugwort at 0, 3, 5 and 10% in concentrate. Four crossbred (CorriedalexPolwarth, t) sheep with a mean body weight of 41.3 kg were used to evaluate nutrients digestibility, palatability, fermentation characteristics and microbial protein synthesis in the rumen. The digestibility of crude protein was improved (p < 0.05) to 6.1 % - 8.6 % in sheep fed 3, 5 and 10% mugwort pellet treatments compared with that of control. That of crude fat and NDF was improved (p < 0.05) to 5.8 % - 7.3 % in sheep fed 3 % compared to other treatments. The ruminal pH was significantly (p < 0.05) decreased in sheep fed 3 % mugwort pellet compared to other treatments when observed at 0.5 hour after feeding. The ammonia nitrogen concentrations were the highest in sheep fed all treatments at 1 hour after feeding. The ruminal concentrations of acetic acid and propionic acids were an improvement (p < 0.05) at the 3% and 5% treatments. Retained nitrogen of 3, 5 and 10% treatment with the value of 2.24 - 2.82 g was higher (p < 0.05) than that of the control with 0.78 g and microbial protein production of 10% treatment was higher (p < 0.05) than that of control. This study suggested that the replacing with 3% dry mugwort (Artemisia montana Pampan) in concentrate a time of preparation pellet will improve nutrient digestibility, palatability, ruminal fermentation characteristics and feed value.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.1
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• pp.51-58
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• 2008

Four ruminally cannulated Nili-ravi buffalo bulls were used in a $4{\times}4$ Latin Square design to determine the influence of varying levels of ruminally degradable protein (RDP) on ruminal characteristics, digestibility, blood pH, blood urea nitrogen (BUN) and nitrogen (N) balance. Four isonitrogenous and isocaloric diets were formulated (NRC, 2001). The control diet contained 50% RDP. The medium (MRDP), high (HRDP) and very high (VHRDP) ruminally degradable protein diets had 66, 82 and 100% RDP, respectively. Increasing the level of dietary RDP resulted in a linear decrease in ruminal pH. A quadratic effect of RDP on ruminal pH was also observed with quadratic maxima at the 66% RDP diet. Dietary RDP had a quadratic effect on total bacterial and protozoal count with maximum microbial count at the 82% RDP diet. Increased microbial count was due to increasing level of ruminal ammonia nitrogen ($NH_3-N$). Increasing dietary RDP resulted in a linear increase in dry matter digestibility. Provision of an adequate amount of RDP caused optimum microbial activity, which resulted in improvement in DM digestibility. Increasing the level of dietary RDP resulted in a linear decrease in crude protein (CP) and neutral detergent fiber digestibility. Blood pH remained unaltered across all diets. A linear increase in ruminal $NH_3-N$ and BUN was noted with increasing level of dietary RDP. The increase in BUN was due to increased ruminal $NH_3-N$ concentrations. A positive N balance was noted across all diets. The results are interpreted to suggest that buffalo bulls can utilize up to 82% RDP of total CP (16%) with optimum results.

• Food Science of Animal Resources
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• v.34 no.3
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• pp.307-315
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• 2014

pH adjustment would be of advantage in improving the water holding capacity of muscle proteins. The objective of this study was to evaluate the addition of fish sarcoplasmic protein (SP) solution, which was adjusted to pH 3.0 or 12.0, neutralized to pH 7.0, and lyophilized to obtain the acid- and alkaline-treated SP samples, on the functional properties of the chicken myofibrillar protein induced by microbial transglutaminase (MTG). The solubility of alkaline-treated SP was higher than that of the acid counterpart; however, those values of the two pH-treated samples were lower than that of normal SP (p<0.05). All SP solutions were mixed with myofibrillar proteins (MP) extracted from chicken breast, and incubated with MTG. The shear stresses of MP with acid- and alkaline-treated SP were higher than that of normal SP. The thermal stability of MP mixture reduced upon adding SP, regardless of the pH treatment. The breaking force of MP gels with acid-treated SP increased more than those of alkaline-treated SP, while normal SP showed the highest value. The MP gel lightness increased, but cooking loss reduced, with the addition of SP. Smooth microstructure of the gel surface was observed. These results indicated that adjusting the pH of SP improved the water holding capacity of chicken myofibrillar proteins induced by MTG.