Rumen microbial growth rates and yields: effect of amino acids and protein.

Effects of amino acids upon microbial growth, optimum ratio of nonprotein to amino acid nitrogen for microbial growth, and incorporation of amino acids into microbial cells were determined with washed cell suspension in vitro as were rumen microbial cells. Rumen microbial dry matter, nitrogen, ribonucleic acid, deoxyribonucleic acid, and substrate disappearance was greatest when a mixture of 18 amino acids was substituted for urea. Substitutions of mixtures of 10 essential amino acids, 8 nonessential amino acids, and sulfur containing amino acids and glutamate also stimulated microbial growth. Acid hydrolyzed casein markedly improved microbial growth. Branched amino acid addition did not affect growth. The optimum ratio of nonprotein to amino acid nitrogen for microbial growth was 75% urea nitrogen and 25% amino acid nitrogen. With this amount of amino acids, an average of 53% of added amino acid was incorporated into microbial cells, 14% was fermented to carbon dioxide and volatile fatty acids, and 33% remained in the supernatant. Both 100% urea and 100% amino acid in growth media were unfavorable for maximal microbial growth. With all carbohydrate substrates, 100% urea nitrogen supported the growth of 9 mg bacterial dry matter per 100 mg of substrate. Substitutions of amino acids for urea increased yields to over 20 mg/100 mg. Microbial growth yields in incubations under carbon dioxide were less than when flasks were flushed with nitrogen. However, yield of bacterial dry matter per unit of substrate was less under nitrogen than under carbon dioxide.     

Factors influencing rumen microbial growth rates and yields: effect of amino acid additions to a purified diet with nitrogen from urea.

Effects of isonitrogenous urea and amino acid additions upon microbial growth in rumen contents from a cow fed a purified diet in which urea was the sole nitrogen source were studied. Incorporation of amino acids into microbial cells, volatile fatty acids, and carbon dioxide was estimated. Rates of microbial growth, volatile fatty acid production, and effects of amino acids upon microbial nitrogen yields were highest right after feeding and decreased with time after feeding. Microbial growth and amounts of amino acids incorporated into microbial cells, volatile fatty acids and carbon dioxide were related closely to quantity of starch remaining in the rumen. High amounts of starch increased microbial protein synthesis from carbon-14 labeled amino acids and reduced amounts of amino acid fermentation. Estimated microbial protein yields per day were 326.0, 444.4, 497.3, and 527.3 g when 0, 15, 30, and 45 mg amino acid nitrogen replaced urea nitrogen during incubation. Respective values for microbial cells per mole estimated adenosine triphosphate were 15.2, 19.2, 21.0, and 24.5. Microbial cell yields per kg carbohydrate digested were 139.0, 189.5, 212.0, and 224.8 g for 0, 15, 30, and 45 mg amino acid nitrogen. Addition of small amounts of amino acids to a diet containing urea as the sole nitrogen source improved considerably rumen microbial protein yields.     

Metabolism of peptides and amino acids during in vitro protein degradation by mixed rumen organisms

In vitro inoculum enriched with particle-associated organisms was prepared using rumen contents from a cow fed a 60% forage, 40% concentrate diet. Treatment of in vitro inoculum with cetyltrimethylammonium bromide was used to release intracellular free amino acids from mixed rumen organisms. Addition of 10 mM tosylarginine methyl ester, a competitive inhibitor of trypsin, decreased degradation rate and intracellular free amino acids in incubations containing either casein or serum albumin. Extracellular peptides increased rapidly to a maximum at 60 min in casein incubations but were not different from zero in albumin incubations. Accumulation of intracellular free amino acids was maximal at 60 min in casein and albumin incubations; the concentration observed with albumin was about one-fourth that with casein. Ammonia production from intact casein was slightly greater than that from acid and enzymatically hydrolyzed casein and about 80% greater than that from albumin. Ammonia production and appearance of extracellular free amino acids lagged behind accumulation of intracellular free amino acids. Results suggest that formation and metabolism of extracellular peptides are important in controlling the rate of protein degradation by mixed rumen organisms.     

Effect of slowly and rapidly degraded protein sources on the concentrations of amino acids and peptides in the rumen of steers

Four steers were given diets of straw and tapioca supplemented with urea, casein, formaldehyde treated casein (FTC), decorticated groundnut meal (DCGM), soya bean meal (SBM), maize gluten meal (MGM) or fish meal (FM) to determine the effect of degradability on rumen amino acid and peptide concentrations. Maximum concentrations of amino acid and peptide-N in rumen ultrafiltrates occurred 1 h after feeding. Total amino acid-N concentrations (mg litre^?1) at this time were low, ranging from 1.2 SE 0.8 (urea) to 15.6 SE 6.0 (casein) with little variation in individual amino acids. The peptide-N concentrations (mg litre^?1) were higher, ranging from 2.4 SE 0.5 (urea) to 160.0 SE 21.0 (casein), but were poorly correlated with degradability and solubility. Apart from casein, the highest concentrations of peptide-N were for those protein supplements, MGM (136.0 SE 25.0), FM (90.0 SE 15.0), and FTC (87.0 SE 14.0), with the lowest degradability suggesting that other factors, such as hydrophobicity, are also involved in the control of rumen peptide concentrations. Pooled samples taken 1 h after the morning feed were separated into three molecular weight (MW) fractions, 5000–50 000 da, 250–5000 da and < 250 da by gel filtration chromatography. From amino acid analysis it was found that the highest proportion of peptide-N was in the 250–5000 da fraction, and although some high MW (5000–50 000) constituents were present, only the FTC and MGM diets gave rise to significant amounts.     

氨酸对嗜酸乳杆菌生长及冻干的影响

探讨了氨基酸对嗜酸乳杆菌生长的影响.采用单因子试验研究了16种氨基酸对嗜酸乳杆菌增菌效果和抗冷冻效应,结果表明,谷氨酸、精氨酸、亮氨酸、赖氨酸、甲硫氨酸、脯氨酸、苯丙氨酸显著促进嗜酸乳杆菌生长,丙氨酸、异亮氨酸、半胱氨酸、丝氨酸、苯丙氨酸、大冬氨酸、甘氨酸、脯氨酸能提高嗜酸乳杆菌抗冷冻能力.研究发现,向发酵液中添加异亮氨酸0.03g/L、半胱氨酸0.03g/L、天冬氨酸0.03g/L、丝氨酸0.03g/L、甘氨酸0.03g/L组成的复合氨基酸或半胱氨酸0.03g/L、天冬氨酸0.03g/L、丙氨酸0.03g/L、甘氨酸0.03g/L组成的复合氨基酸,均可使嗜酸乳杆菌冻干存活率达到95%以上,而对照组只有42.95%.     

Effect of diet on amino and nucleic acids of rumen bacteria and protozoa

Amino acid composition and nucleic acid content of pure cultures of rumen bacteria (17 species) were analyzed. Amino acid composition between gram-positive and -negative organisms was not different. The total nitrogen content of gram-negative bacteria (10.8%) was significantly higher than gram-positive organisms (9.9%). Deoxyribonucleic acid-nitrogen: total nitrogen (mg/g) differed between gram-positive (8.8) and gram-negative (18.9) bacteria, but there was no significant difference in ratio of ribonucleic acid-nitrogen to total nitrogen. In a second experiment six rumen-fistulated cattle were fed either a high roughage (85% alfalfa hay plus 15% concentrate) or high concentrate diet (15% alfalfa hay and 85% concentrate). Cattle were adapted 14 days and rumen contents sampled on 3 consecutive days. Nitrogen content was higher in protozoa from cattle fed low concentrate (8.4%) than in protozoa from cattle fed high concentrate (7.9%) but was similar in bacteria for both diets. Deoxyribonucleic acid nitrogen: total nitrogen (mg/g in bacteria decreased from 27.2 in cattle fed the low concentrate diet to 20.9 in those fed the high concentrate diet. Differences between sampling days were significant for both bacteria and protozoa for ratio of deoxyribonucleic acid-nitrogen to total nitrogen but were significant only in protozoa for ratio of ribonucleic acid nitrogen to total nitrogen. Ribonucleic acid may serve as a marker for estimating microbial production in the rumen if sources of variation are recognized and corrected adequately.     

氨基酸和肽在食品中的呈味作用

本文综述了在食品中有呈味作用的氨基酸和肽。天然氨基酸大多具有甜味或苦味,MSG有强烈的鲜味,阿斯巴甜、阿力甜、索马钳和莫奈林是强力甜味剂。鲜味二肽、咸味二肽、苦味肽、美味肽等也出现了潜在的商业价值。     

Effects of fiber content and particle size of forage on the flow of microbial amino acids from continuous culture fermenters

Eight dual-flow continuous culture fermenters (1320 mL) were used in two 10-d consecutive periods to study the effects of fiber content and particle size on rumen fermentation, nutrient flow, and the profile and flow of amino acids (AA) from microbial origin. Treatments were arranged in a 2 x 2 factorial; the main factors were fiber content (high fiber [HF] = 67% alfalfa hay, 33% concentrate; low fiber [LF] = 39% alfalfa hay, 61% concentrate) and forage stem particle size (ground not to pass a 3-mm sieve = large size [LS]; ground to pass a 1-mm sieve = small size [SS]). On the last day of the experiment, liquid- (LAB) and solid- (SAB) associated bacteria were isolated from each fermenter for chemical analysis. Microbial N and AA flows were estimated using LAB or SAB composition data. Total volatile fatty acid concentration was higher for LF (122 mM) than for HF (102 mM). The proportion of acetate was higher for HF (63.5%) than for LF (58.2%). When SS was fed instead of LS, there was a reduction in the acetate proportion (63.4% vs 58.3%, respectively) and in the acetate to propionate ratio (3.13 vs. 2.41, respectively). Bacterial N flow was higher, and dietary N flow was lower, in SS compared with LS when LAB were used for calculations. Efficiency of microbial protein synthesis (EMPS) was affected by particle size, when LAB were used for calculations, and by fiber content, when SAB were used for calculations. The AA profile of LAB differed from SAB in 4 of 16 AA. The fiber content and particle size had small effects on microbial AA profile. However, the bacterial AA flow was higher in 13 of 16 AA, when SAB instead of LAB were used for calculations. Estimates of bacterial N flow, EMPS, AA profile, and flows of microbial origin were dependent on the type of microbial population used for calculations. The representativeness of the microbial samples is essential for the accurate prediction of the AA flow from microbial origin.     

Correlation of bitterness thresholds of amino acids and peptides with molecular connectivity

Molecular connectivity values are easily calculated topological indices related to the degree of branching (hence the size and shape) of any chemical structure considered. These structural parameters are significantly correlated with literature values for the bitterness thresholds of amino acids, peptides and their derivatives. The quantitative correlation of flavour thresholds with structural parameters may prove useful for predicting thresholds of untested compounds; the potential and disadvantages of this approach are discussed.     

Dynamics of fermentation of a purified diet and microbial growth in the rumen.

Ruminal fermentation and disappearance of glucose, starch, and cellulose, and incorporation of glucose and starch into microbial cells were estimated in a fistulated Jersey cow fed twice daily a purified diet containing urea as the sole nitrogen source. Estimated rumen volume was 59.8 liters. Turnover time and rate of passage of rumen contents were 33.4 h and 1.8 liters per h. Turnover times of glucose, starch, and cellulose were .17, 4.7, and 14.2 h. Fermentation times of glucose, starch, and cellulose were .17, 5.5, and 25.1 h. Percentages of glucose, starch, and cellulose utilized in the rumen were 99.4, 85.4, and 60.6. Thus, 18.5% of the carbohydrate fed bypassed rumen fermentation, and 81.5% was utilized in the rumen. All glucose disappeared from the rumen within an hour. An average of 32.1, 43.0, and 14%, respectively, of glucose utilized was incorporated into microbial cells, volatile fatty acids, and carbon dioxide. Percentage of starch incorporated into cells varied, with time being highest 2 h after feeding at 40% and lowest at 20%, 10 h after feeding. Respective percentages of starch incorporated into microbial cells, volatile fatty acids, and carbon dioxide were 32.4, 45.9; and 13.3. Total microbial protein and cell yields per kilogram carbohydrate utilized in the rumen were 77.1 and 117.5 g. Microbial cell yield per mole (estimated) of adenosine triphosphate was 16.2 g.     

富含硫氨基酸转基因大豆对根际土壤微生物群落结构的影响

选用两组转基因品质改良大豆（富含硫氨基酸）,利用磷脂脂肪酸（PLFA）法分析了在大田种植条件下,两组转基因大豆对土壤微生物群落结构的影响。结果表明,与非转基因大豆相比,转基因品系根际土壤微生物群落的结构发生明显的变化。在两组转基因品系中PLFA组分均发生变化,A组中检测到15种特异性的PLFA,其中占主导地位的分别是是革兰氏阳性菌（GP）的特征脂肪酸16∶0、10Me18∶0和好氧细菌特征脂肪酸18∶1ω7c;B组中检测到13种特异性的PLFA,其中占主导地位的是表征GP的脂肪酸15∶0、16∶0和非特征脂肪酸14Me16∶0。在A组的OE-8品系及B组的17-4和57品系中,表征细菌的PLFA含量和总PLFA含量与非转基因品系相比有显著上升。结果表明种植转基因大豆影响根际土壤微生物群落结构。     

不同起垄时间对烟株长势及烟叶产质量的影响

本试验主要针对山东烟区春季干旱少雨的自然生态条件,以推广冬起垄或早春起垄盖膜为目的,通过对冬起垄或早春起垄的研究,确定最佳起垄时间.结果表明,封冻前起垄及冬耕后及时起垄烟田效果较好,能够达到集蓄冬季雨雪、缓解春季干旱、提高烟叶产量和质量的目的.     

Effects of corn processing on growth characteristics, rumen development, and rumen parameters in neonatal dairy calves

Neonatal Holstein calves were fed texturized calf starters containing 33% whole (WC), dry-rolled (DRC), roasted-rolled (RC), or steam-flaked (SFC) corn to investigate how corn processing method affects intake, growth, rumen and blood metabolites, and rumen development. In the first experiment, 92 Holstein calves (52 male and 40 female) were started at 2 +/- 1 d of age and studied for 42 d. Starter dry matter (DM) intake was measured and fecal scoring conducted daily. Growth and blood parameter measurements were conducted weekly. A subset of 12 male calves (3/treatment) was euthanized at 4 wk of age and rumen tissue sampled for rumen epithelial development measurements. Experiment 2 consisted of 12 male Holstein calves ruminally cannulated at 7 +/- 1 d of age. Rumen fluid and blood samples were collected during wk 2 to 6. In the first experiment, postweaning and overall starter and total DM intake were significantly higher in calves fed starter with DRC than RC or SFC. Postweaning and overall starter and total DM intake were significantly higher in calves fed starter with WC than SFC. Postweaning average daily gain was significantly greater in calves fed starter with DRC than SFC. Blood volatile fatty acid concentrations were significantly higher in calves fed starter with SFC than in calves fed all other treatments. Papillae length and rumen wall thickness at 4 wk were significantly greater in calves fed starter with SFC than DRC and WC, respectively. In experiment 2, calves fed starter with WC had higher rumen pH and lower rumen volatile fatty acid concentrations than calves fed all other starters. Rumen propionate production was increased in calves receiving starter with SFC; however, rumen butyrate production was higher in calves fed starter with RC. Results indicate that the type of processed corn incorporated into calf starter can influence intake, growth, and rumen parameters in neonatal calves. Calves consuming starter containing RC had similar body weight, feed efficiency, and rumen development but increased structural growth and ruminal butyrate production when compared with the other corn processing treatments.     

Use of odd and branched-chain fatty acids in rumen contents and milk as a potential microbial marker

The objectives of this study were: 1) to determine if a correlation exists between rumen odd and branched-chain fatty acids (OBCFA, i.e., C(15:0), iso C(15:0), anteiso C(15:0), C(17:0), iso C(17:0), anteiso C(17:0), and C(17:1)), uracil, and purine bases (PB), 2) to evaluate the potential of milk OBCFA secretion to predict duodenal flow of microbial protein in lactating cows, 3) to evaluate the accuracy of the latter prediction equations using an independent data set, and 4) to determine whether these predictions were more accurate than predictions based on dry matter intake (DMI) and dietary characteristics. In the first experiment, 4 lactating dairy cows arranged in a 4 x 4 Latin square were offered diets based on grass silages of different botanical composition and a standard concentrate. The relationship between rumen pool size of OBCFA and microbial matter was investigated. Rumen pool size of microbial matter (g), determined 4, 12, and 17 h after feeding, using uracil and PB as microbial markers, was closely related to OBCFA (g) [r(2) = 0.716, root mean square error (RMSE) = 4.45]. To correct for differences in marker concentrations among strains of rumen microbes, C(17:0) was included in the regression equations, resulting in an increased predictive power (r(2) = 0.780, RMSE = 3.92). The relationship between microbial flow to the duodenum and milk OBCFA yield was evaluated in a second experiment with lactating dairy cows offered diets based on grass silage and concentrates differing in starch source. Similar to observations in the rumen, milk OBCFA yield was closely related to microbial flow to the duodenum (RMSE = 4.28), but predictive power of equations did not increase when straight-chain C(17)-fatty acids were included in the regression equations (RMSE = 4.92). Evaluation of the current prediction equations with 3 independent datasets resulted in a root mean square prediction error of 20.5 and 13.4% of the observed mean for equations based on milk secretion of total OBCFA and straight-chain C(17)-fatty acids, respectively. Comparison of the accuracy of the latter equations with 2 previously published equations based on DMI and dietary characteristics suggest the former to be more accurate. This first evaluation suggests that milk OBCFA could be used as a marker for duodenal flow of microbial matter, especially when accurate measurements of DMI are not available.     

Effects of cinnamaldehyde and garlic oil on rumen microbial fermentation in a dual flow continuous culture

Eight continuous culture fermentors inoculated with ruminal liquor from heifers fed a 50:50 alfalfa hay:concentrate diet (17.6% crude protein, 28.0% neutral detergent fiber) were used in 3 replicated periods to study the effects of cinnamaldehyde (CIN) and garlic oil (GAR) on rumen microbial fermentation. Treatments were no additive (negative control); 1.25 mg/L (MON) and 12.5 mg/L (MON10) of the ionophore antibiotic monensin (positive control); 31.2 mg/L CIN (CIN) and 312 mg/L (CIN10) of CIN; and 31.2 mg/L GAR (GAR) and 312 mg/L (GAR10) of GAR (Allium sativa). The MON10 caused expected changes in microbial fermentation patterns (a decrease in fiber digestion, ammonia N concentration, and proportions of acetate and butyrate; an increase in the proportion of propionate; and a trend to increase small peptide plus AA N concentration). The CIN decreased the proportion of acetate and branch-chained volatile fatty acids (VFA) and increased the proportion of propionate; CIN10 decreased the proportion of acetate and increased the proportion of butyrate compared with the control. The GAR10 increased the proportion of propionate and butyrate and decreased the proportion of acetate and branch-chained VFA compared with the control. The GAR10 also increased the small peptide plus amino acid N concentration, although no effects were observed on large peptides or ammonia N concentrations. The CIN and GAR10 resulted in similar effects as monensin, with the exception of the effects on the molar proportion of butyrate, which suggests that they might have a different mode of action in affecting in vitro microbial fermentation.     

Influence of Kluyveromyces marxianus on proteins,peptides, and amino acids in Lactobacillus-fermented milk

Food Science and Biotechnology - With increasing application of yeast in fermented milk, in order to study the effect of yeast on milk protein during the fermentation process, the effects of the...     

抗氧化剂对甘氨酸—葡萄糖体系中氨基酸的影响

采用葡萄糖与甘氨酸在160℃下反应建立美拉德反应的模拟体系,通过离子色谱测定其氨基酸的种类与含量,探讨了四种抗氧化剂BHA、BHT、TBHQ、维生素C对美拉德反应产物中氨基酸的影响.结果表明,添加抗氧化剂对葡萄糖-甘氨酸体系中氨基酸的种类与含量都有显著影响,其中添加BHT产生的酪氨酸最多,添加维生素C产生的组氨酸最多,添加TBHQ、BHT产生了精氨酸,添加TBHQ、维生素C产生了赖氨酸,添加BHT还产生了丝氨酸,添加TBHQ还产生了亮氨酸.而单独BHA与甘氨酸、维生素C与甘氨酸油浴后都检测到甘氨酸与组氨酸,BHT、TBHQ与甘氨酸油浴后只检测到甘氨酸.但是单独抗氧化剂与葡萄糖油浴后都没有检测到任何氨基酸,说明氨基酸是在美拉德反应中产生的,而且添加不同种类抗氧化剂对美拉德反应产物中氨基酸的种类与含量会产生显著影响.     

Effect of direct-fed microbials on rumen microbial fermentation.

Nonbacterial, direct-fed microbials added to ruminant diets generally consist of Aspergillus oryzae fermentation extract, or Saccharomyces cerevisiae cultures, or both. Results from in vivo research have been variable regarding effects of direct-fed microbials on ruminant feedstuff utilization and performance. Some research has shown increased weight gains, milk production, and total tract digestibility of feed components, but others have shown little influence of direct-fed microbials on these parameters. In vitro research with mixed ruminal microorganisms likewise has been inconsistent regarding the effects of direct-fed microbials. Several researchers observed that direct-fed microbials increased cellulolytic bacterial numbers in the rumen and stimulated the production of some fermentation end products. This suggests that direct-fed microbials may be providing growth factors for the ruminal microbes. However, other researchers have reported no effect of direct-fed microbials on in vitro fiber digestion. Recent research demonstrated that growth of the predominant ruminal bacterium Selenomonas ruminantium in lactate medium as well as lactate uptake by whole cells of Sel. ruminantium were markedly increased by an A. oryzae fermentation extract and an S. cerevisiae culture. In addition, both products increased the production of acetate, propionate, succinate, total VFA, and cell yield (grams of cells per mole of lactate). Therefore, it appears that these direct-fed microbials provide soluble factors that stimulate lactate utilization by Sel. ruminantium. Evidence is presented indicating that the malate content of the A. oryzae fermentation extract and S. cerevisiae culture may be involved in this stimulation.     

有机活性剂对烤烟生长和产量及质量的影响

2003年利用田间试验研究了3种烟草专用有机活性剂对烤烟生长、产量和质量的影响.结果表明,有机活性剂能够提高烟株的生理活性,促进烤烟生长,明显提高烤烟各部位叶面积的扩展,对烤烟产量和产值的提高达到显著或极显著水平,烤烟的内在品质也得到明显提高.3种活性剂综合比较,2号活性剂的效果相对更好.