This study aimed to evaluate effects of 800 W microwave irradiation for 2, 4 and 6 min on chemical composition, antinutritional factors, ruminal dry matter (DM) and crude protein (CP) degradability, and in vitro CP digestibility of canola seed (CS). Nylon bags of untreated or irradiated CS were suspended in the rumen of three bulls from 0 to 48 h. Protein subfractions of untreated and microwave irradiated CS before and after incubation in the rumen were monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Microwave irradiation had no effect on chemical composition of CS (p>0.05). There was a linear decrease (p<0.001) in the phytic acid and glucosinolate contents of CS as irradiation time increased. Microwave irradiation for 2, 4 and 6 min decreased the phytic acid content of CS by 8.2, 27.6 and 48.6%, respectively. The total glucosinolate contents of CS microwave irradiated for 2, 4 and 6 min decreased by 41.5, 54.7 and 59.0% respectively, compared to untreated samples. The washout fractions of DM and CP and degradation rate of the b fraction of CP decreased linearly (p<0.001) as irradiation time increased. Microwave irradiation for 2, 4 and 6 min decreased effective degradability (ED) of CP at a ruminal outflow rate of 0.05 $h^{-1}$ by 4.7, 12.3 and 21.0%, respectively. Microwave irradiation increased linearly (p<0.001) in vitro CP digestibility of ruminally undegraded CS collected after 16 h incubation. Electrophoresis results showed that napin subunits of untreated CS disappeared completely within the zero incubation period, whereas cruciferin subunits were degraded in the middle of the incubation period (16 h incubation period). In 4 and 6 min microwave irradiated CS, napin subunits were degraded after 4 and 16 h incubation periods, respectively, and cruciferin subunits were not degraded untile 24 h of incubation. In conclusion, it seems that microwave irradiation not only protected CP of CS from ruminal degradation, but also increased in vitro digestibility of CP. Moreover, microwave irradiation was effective in reducing glucosinolate and phytic acid contents of CS.
Four ruminally fistulated Hanwoo steers were used to determine the effects of level and degradability of dietary protein on ruminal fermentation, blood metabolites and concentration of soluble non-ammonia nitrogen (SNAN) in ruminal (RD) and omasal digesta (OD). Experiments were conducted in a $4{\times}4$ Latin square design with a $2{\times}2$ factorial arrangement of treatments. Factors were protein supplements with two ruminal crude protein (CP) degradabilities, corn gluten meal (CGM) that was low in degradability (rumen-degraded protein (RDP), 23.4% CP) or soybean meal (SBM) that was high in degradability (RDP, 62.1% CP), and two feeding levels of CP (12.2 or 15.9% dry matter). Ruminal fermentation rates and plasma metabolite concentrations were determined from the RD collected at 2-h intervals and from the blood taken by jugular puncture, respectively. The SNAN fractions (free amino acid, peptide and soluble protein) in RD and OD collected at 2-h intervals were assessed by ninhydrin assay. Mean ruminal ammonia concentrations were 40.5, 74.8, 103.4 and 127.0 mg/L for low CGM, high CGM, low SBM and high SBM, respectively, with statistically significant differences (p<0.01 for CP level and p<0.001 for CP degradability). Blood urea nitrogen concentrations were increased by high CP level (p<0.001) but unaffected by CP degradability. There was a significant (p<0.05) interaction between level and degradability of CP on blood albumin concentrations. Albumin was decreased to a greater extent by increasing degradability of low CP diets (0.26 g/dl) compared with high CP diets (0.02 g/dl). Concentrations of each SNAN fraction in RD (p<0.01) and OD (p<0.05) for high CP diets were higher than those for low CP diets, except for peptides but concentrations of the sum of peptide and free amino acid in RD and OD were significantly higher (p<0.05) for high CP diets than for low CP diets. Soybean meal diets increased free amino acid and peptide concentrations in both RD (p<0.01) and OD (p<0.05) compared to CGM diets. High level and greater degradability of CP increased (p<0.001) mean concentrations of total SNAN in RD and OD. These results suggest that RDP contents, increased by higher level and degradability of dietary protein, may increase release of free amino acids, peptides and soluble proteins in the rumen and omasum from ruminal degradation and solubilization of dietary proteins. Because SNAN in OD indicates the terminal product of ruminal metabolism, increasing CP level and degradability appears to increase the amount of intestine-available nitrogen in the liquid phase.
An experiment was conducted as a Latin square design with four rumen fistulated local yellow cattle with a mean live weight of 230 kg. The treatments were: $(CLM_0)$ urea-treated rice straw ad libitum plus 1 kg cassava root meal (basal diet), $(CLM_{500})$ basal diet plus 500 g cassava leaf meal, $(CLM_{1000})$ basal diet plus 1,000 g cassava leaf meal, and $(CLM_{1500})$ basal diet plus 1,500 g cassava leaf meal. The results showed that there were differences in dry matter intake of urea-treated rice straw between treatments (p<0.05). The highest total dry matter intake was observed for treatment $CLM_{1500}$, with 2.62 kg DM/100 kg LWt/day, followed by treatments $CLM_{1000}$, $CLM_{500}$ and $CLM_0$, with 2.42, 2.00 and 1.86 kg DM/100 kg LWt/day, respectively. The ruminal ammonia concentration on treatment $CLM_{1500}$ was greater than on treatments $CLM_{1000}$, $CLM_{500}$ and $CLM_0$. There were non-significant differences in the ruminal pH among the treatments. The in sacco degradability of cassava leaf meal and cassava root meal was high, and on average 75 and 85% respectively of the DM had disappeared after 24 h of incubation. Degradation rate of urea treated rice straw was 64% after 72 h of incubation.
The aim of this study was to predict the energy value and dynamic degradation of roughage in Taiwan using the $Daisy^{(R)}$. in vitro fermentation method to provide information on one of the very important nutrients for ration formulation. The second objective was to study the effects of Aspergillus oryzae (AFE) inclusion on nutrient utilization. Three ruminal fistulated dry dairy cows were used for rumen fluid and fifteen conventional forages used in dairy cattle were collected around this island. The degradability of these feedstuffs with and without AFE ($Amaferm^{(R)}$.) treatment was measured using the $Daisy^{(R)}$. in vitro method. The roughage energy values, including TDN and NEL, were calculated according to Robinson (2000). Results from the 30 h in vitro neutral detergent fiber (NDF) degradability and predicted energy evaluations showed that alfalfa (among the forages) contained the highest degradability and energy values, Bermuda straw having the lowest. Peanut vines and corn silage contained higher energy values and the lowest value found in Pangola and Napier grasses among the locally produced forages. Pangola and Napier grasses had lower values than most imported forages except Bermuda straw. Among the by-products, wheat middling contained the highest NDF degradability, while rice bran contained the richest energy value due to its high oil content. From the dynamic dry matter (DM), organic matter (OM), acid detergent fiber (ADF) and neutral detergent fiber (NDF) degradation, corn silage contained the highest effective degradation among the local forages; wheat middling (among the by-products) degraded the fastest in DM, OM, ADF and NDF and showed the highest effective degradability. AFE inclusion was inconsistent among the forages. Alfalfa hay showed significantly increased 30 h NDF degradability and energy values, Pangola hay, Napier grass and brewer's grains showed decreased degradability and energy values. AFE inclusion increased the DM, OM and NDF degradation rate in most forage, but only increased the DM degradation rate in sorghum distiller's grains, the OM degradation rate in bean curd pomace and the NDF and ADF degradation rates in soy pomace (among the by-products).
The objective of this study was to examine the impact of three categories of supplements on intake and diet induced difference on degradation of straw substrates. Sixteen crossbred cattle fitted with rumen cannula were randomly divided into 4 equal groups. Animals were fed on wheat straw ad libitum without any supplement except mineral mixture (control; $T_1$) or supplemented with concentrate mixture (CS; $T_2$) or green Lucerne (GLS; $T_3$) or urea-molasses block lick (ULS; $T_4$). Total dry matter intake in $T_2$, $T_3$ and $T_4$ was increased by 70, 54 and 49%, respectively compared to $T_1$ which was only 1.55 kg/100 kg B.Wt. Other than control animals, straw intake was less on $T_3$ than $T_2$ or $T_4$. In Sacco degradation of untreated and urea treated wheat or paddy straw in different treatments indicated that the supplements had a significant (p<0.01) impact on rapidly soluble (A) and insoluble but potentially degradable (B) fractions of straw. Urea treatment increased fraction-A but, provision of supplement improved fraction-B also. Effective degradation (ED) of OM was better on $T_2$. Rate of degradation (C) of OM and CWC was dependent on diet and type of straw but hemicellulose and cellulose were related to latter factor only. ED of cell wall carbohydrates (CWC) was similar in $T_2$ and $T_4$ but higher than $T_3$. CS was more effective in improving the degradation of both untreated and urea treated straw while ULS was effective on the former only. CS had more impact on superior quality straw while contrary was true with ULS. Although GLS improved intake and degradability of untreated and urea treated straws, its bulkiness affected the straw intake compared to other supplements.
The effect of maize hybrid (Suco and Dekalb 765, DK 765), maturity stage (milk, $R_3$ and 1/2 milk line, $R_5$) and animal diet (Diet 1: 70% lucerne hay+30% maize silage; Diet 2: 50% maize silage+20% sunflower meal+30% maize grain) on ruminal stover dry matter (DM) degradability was studied. Additionally, morphological and chemical plant composition was evaluated. Fodder samples ground at 2 mm were incubated in three Holstein steers (400 kg body weight) using the in situ technique. Ruminal degradation kinetics was studied and the effective degradability (ED) was estimated for an assumed kp of 5%/h. The in situ data was analyzed in a complete randomized block design with the animals as blocks. Significant interactions between hybrid${\times}$diet and maturity${\times}$diet on kinetic digestion parameters were detected. In Diet 1, hybrids did not differ in degradable fraction, kd or ED, although a minor difference (p<0.05) in the soluble fraction was found (25.5 and 23.2% for Suco and DK 765, respectively). In Diet 2, the DK 765 had greater degradable fraction (p<0.001) but smaller (p<0.01) kd than Suco, without differences in the soluble fraction or in ED. Anticipating the harvest increased ED of stover from 37.5% in $R_5$ to 44.6% in $R_3$ (average values across hybrids and diets) due to the increase (p<0.001) in the soluble fraction ($R_5$: 22.6%, $R_3$: 28.8%). It is concluded that hybrids had similar stover in situ DM degradability and that soluble fraction represent the main proportion of degradable substrates. Advancing the date of harvesting may not improve the in situ DM degradability of whole maize plant silage since the increase in stover quality is counteracted by the depression in the grain-to-stover ratio. The diet of the animal consuming silage might not improve stover utilization either.
In vivo digestion and metabolism trials were conducted with 10 wethers equipped with ruminal, abomasal, and ileal cannulae to evaluate digestion of ensiled broiler litter (EBL), deepstacked broiler litter (DBL), and composted broiler litter (CBL). Wethers were fed a low protein (6.3% CP) basal diet alone or supplemented to 10.3% CP with EBL, DBL, CBL or soybean meal (SBM). All diets were formulated to be isoenergetic (56% TDN, DM basis). Apparent digestibilities of DM, OM, and ADF were not affected (p<0.05) by diet, but digestibility of CP was improved (p<0.05) by N supplementation. Apparent digestibility of CP was lower (p<0.05) for diets supplemented with CBL and DBL than for diets supplemented with SBM and EBL. Ruminal $NH_3$ concentration was 20 to 24 mg/dl at 2 h after feeding litter-supplemented diets compared with 13 mg/dl for SBM. Abomasal N, $NH_3$ N, and nonammonia N flows were increased (p<0.05) by N supplementation, whereas microbial N flow was not influenced (p<0.05) by diet. Compared with SBM and EBL, undegraded dietary CP flow to the abomasum tended to be greater (p<0.1) when wethers were fed DBL and CBLsupplemented diets. Retention of N (g/d) also was greater (p<0.05) due to greater (p<0.05) N intake and lower (p<0.05) urinary N excretion when wethers were fed diets supplemented with litter (especially EBL) vs. SBM. Overall, characteristics of ruminal fermentation and digestion indicated that broiler litter N was utilized efficiently by wethers, but ensiling may be preferable to deepstacking or composting.
An experiment was conducted to study the effect of soluble protein supplements on concentration of soluble non-ammonia nitrogen (SNAN) in the liquid phase of ruminal (RD) and omasal digesta (OD) of Korean native steers, and to investigate diurnal pattern in SNAN concentration in RD and OD. Three ruminally cannulated Korean native steers in a $3{\times}3$ Latin square design consumed a basal diet of rice straw and corn-based concentrate (control), and that supplemented (kg/d DM basis) with intact casein (0.24; IC) or acid hydrolyzed casein (0.46; AHC). Ruminal digesta was sampled using a vacuum pump, whereas OD was collected using an omasal sampling system at 2.0 h intervals after a morning feeding. The SNAN fractions (free amino acid (AA), peptide and soluble protein) in RD and OD were assessed using the ninhydrin assay. Concentrations of free AA and total SNAN in RD were significantly (p<0.05) lower than those in OD. Although free AA concentration was relatively high, mean peptide was quantitatively the most important fraction of total SNAN in both RD and OD, indicating that degradation of peptide to AA rather than hydrolysis of soluble protein to peptide or deamination may be the most limiting step in rumen proteolysis of Korean native steers. Diurnal variation in peptide concentration in OD for the soluble protein supplemented diets during the feeding cycle peaked 2 h post-feeding and decreased thereafter whereas that for the control was relatively constant during the entire feeding cycle. Diurnal variation in peptide concentration was rather similar between RD and OD.
Responses of the rumen fungus, Neocallimastix frontalis RE1, to long chain fatty acid (LCFA) were evaluated by measuring gas production, filter paper (FP) cellulose digestion and polysaccharidase enzyme activities. LCFA (stearic acid, $C_{18:0}$; oleic acid, $C_{18:1}$; linoleic acid, $C_{18:2}$ and linolenic acid, $C_{18:3}$) were emulsitied by ultrasonication under anaerobic condition, and added to the medium. When N frontalis RE1 was grown in culture with stearic, oleic and linoleic acid, the cumulative gas production, gas pool size, FP cellulose digestion and enzymes activities significantly (p<0.05) increased at some incubation times(especially, exponential phases of fungal growth, 48~120 h of incubation) relative to that for control cultures. However, the addition of linolenic acid strongly inhibited all of the investigated parameters up to 120 h incubation, but not after 168 and 216 h of incubation. These results indicated that stearic, oleic and linoleic acids tended to have great stimulatory effects on fungal cellulolysis, whereas linolenic acid caused a significant (p<0.05) inhibitory effects on the cellulolysis by the rumen fungus. These results are the first report of the effect of LCFAs on the ruminal fungi. Further research is needed to identify the mode of action of LCFAs on fungal strains and to verify whether or not ruminal fungi have ability to hydrate unsaturated LCFAs to saturated FAs. There was high correlation between cumulative in vitro gas production and fungal growth (94.78%), FP cellulose degradation (96.34%), CMCase activity(90.86%) or xylanase activity (87.67%). Thus measuring of cumulative gas production could be a useful tool for evaluating fungal growth and/or enzyme production by ruminal fungi.
Whole cotton seed (WCS) has become one of the major feed ingredients in TMR for dairy cattle in Korea, and WCS for feed use is mostly imported from abroad. Since this genetically modified oil seed is usually fed to the animal in raw state, its germination ability, if last long, often causes concerns about ecological disturbances. In the process of looking for effective conditions to remove germination ability of WCS this study had the objectives to evaluate the nutritional effects of gamma irradiation at doses of 8, 10 and 12 kGy on changes in nutrient contents, anti-nutritional factors, in vitro digestibility and ruminal degradability. No significant differences were found in proximate analysis of nutrients between raw WCS and gamma irradiated one. Glycine and threonine contents significantly increased when the WCS was exposed to gamma ray as compared to untreated WCS (p<0.05). As for fatty acid composition, no significant differences were observed with the irradiation treatment. Free gossypol in WCS was decreased (p<0.05) by gamma irradiation treatment. Of the 3 different levels of gamma irradiation, a dose of 12 kGy was found to be the most effective in reducing free gossypol concentration. Results obtained from in situ experiment indicated that gamma irradiation at a dose of 10 kGy significantly (p<0.05) lowered rumen degradability of both dry matter and crude protein as compared with raw WCS. However, there were no significant differences in rapidly degradable and potentially degradable fractions of crude protein due to 10 kGy gamma irradiation. Overall, this study show that gamma irradiation at a dose of 10 kGy is the optimum condition for removing germination ability of WCS, and could improve nutritive value for the ruminant with respect to the decrease in both ruminal protein degradability and gossypol content of WCS.
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