Methylation methods for the quantitative analysis of conjugated linoleic acid (CLA) isomers in various lipid samples

Precise methylation methods for various chemical forms of conjugated linoleic acid (CLA), which minimize the formation of t,t isomers and allylmethoxy derivatives (AMD) with the completion of methylation, were developed using a 50 mg lipid sample, 3 mL of 1.0 N H(2)SO(4)/methanol, and/or 3 mL of 20% tetramethylguanidine (TMG)/methanol solution(s). Free CLA (FCLA) was methylated with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). CLA esterified in safflower oil (CLA-SO) was methylated with 20% TMG/methanol (100 degrees C, 5 min), whereas CLA esterified in phospholipid (CLA-PL) was methylated with 20% TMG/methanol (100 degrees C, 10 min), followed by an additional reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). Similarly, CLA esterified in egg yolk lipid (CLA-EYL) was methylated by base hydrolysis, followed by reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). These results suggest that for the quantitative analysis of CLA in lipid samples by GC, proper methylation methods should be chosen on the basis of the chemical forms of CLA in samples.     

Structure of a new echinocystic acid bisdesmoside isolated from Codonopsis lanceolata roots and the cytotoxic activity of prosapogenins

We isolated a new saponin named codonoposide (1) from the roots of Codonopsis lanceolata (Campanulaceae) and characterized it as 3-O-[beta-D-xylopyranosyl(1-3)-beta-D-glucuronopyranosyl]-3beta,16alpha-dihydroxyolean-28-oic acid 28-O-[beta-D-xylopyranosyl (1-3)-alpha-L-rhamnopyranosyl (1-2)-alpha-L-arabinopyranosyl] ester by chemical, physicochemical, and 2DNMR techniques. Complete hydrolysis of 1 produced a sapogenin (1a), and the partial hydrolysis and further isolation afforded two prosapogenins (1b, 1c). The structures of 1a, 1b, and 1c were found to be 3beta,16alpha-dihydroxyolean-28-oic acid (echinocystic acid, 1a), 3-O-beta-D-glucuronopyranoside of 1a, and 3-O-beta-D-xylopyranosyl (1-3)-beta-D-glucuronopyranoside of 1a, respectively, on the basis of spectroscopic data. On MTT assay, 1a showed marginal cytotoxic activity whereas 1b exhibited more cytotoxicity than 1a. However, the bisdesmosylsaponin 1 exhibited no cytotoxicity (IC(50)>0.3 mM against tested cell lines). This result indicated that glycoside linkage of glucuronic acid at C-3 enhances the cytotoxicity of sapogenin (1a), and additive glycosylation of xylose to 1b strongly enhances the cytotoxicity of 3-O-monosaccharides (1b). Therefore, true forms of codonoposide for the cytotoxicity must be sapogenins or prosapogenins.     

Influence of Oil Seed Rape and Faba Bean as Green Manure on Nitrogen Utilization in a Spring Cereal Rotation

A lysimeter study was conducted to investigate how nitrogen uptake of spring wheat and oats and N leaching losses are differing when either oilseed rape or faba bean are grown as green manure. The experiment was conducted during 1982–1988, and fertilizer applied to the cereals was either farmyard manure, FYM, (6 and 12 g N/m²/year) or mineral fertilizer, MiF, at a rate of 6 g N/m². On average of the 7 year period faba bean increased N uptake of the cereals fertilized with FYM at harvest by 3–4 g/m². Differences between the two application rates were negligible. N-leaching losses increased twofold when faba bean was grown instead of oilseed rape. Growing faba bean after cereals supplied with MiF, too, increased N uptake of the cereals and significantly increased leaching losses.     

Effects of dietary fiber of young adult genetically lean, obese and contemporary pigs: rate of passage, digestibility and microbiological data

Twenty-one genetically lean, obese or contemporary slaughter weight castrated male pigs (6 mo old; seven of each genotype) were assigned to individual tether stalls and fed either a control diet (low fiber) or a diet containing 80% alfalfa meal (high fiber) at 1.50% of initial body weight for 71 d (1.75% for d 1 to 4). Apparent dry matter digestibility of the diets was estimated by determining acid insoluble ash in fecal samples. Fecal cellulolytic bacteria and total viable bacteria were enumerated at d 0, 14, 35, 49 and 70. Fecal inocula were used to determine 48-h in vitro digestibility of alfalfa meal fractions on the same days. Digesta rate of passage was determined by feeding a pulse dose of chromium-mordanted alfalfa fiber to the pigs fed the high-fiber diet. In vivo digestibility of both diets was less for the obese pigs than for the lean or contemporary genotypes. In vitro digestibility of alfalfa fiber fractions was not different between the genotypes fed either diet. When the high-fiber diet was fed, in vitro digestibility increased for all genotypes from d 0 to d 14, but not thereafter. The numbers of cellulolytic bacteria for all three genotypes were greater when pigs were fed the high-fiber diet (23.0 X 10(8), 51.6 X 10(8), 37.2 X 10(8) per gram fecal dry weight; obese, lean and contemporary, respectively) compared to the low-fiber diet (3.0 X 10(8), 3.2 X 10(8), 3.4 X 10(8), respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of dietary fiber on young adult genetically lean, obese and contemporary pigs: body weight, carcass measurements, organ weights and digesta content

Twenty-one genetically lean, obese or contemporary barrows (6 mo old; seven of each genotype) were assigned to individual tether stalls and fed a control diet (low-fiber) or a diet containing 80% alfalfa meal (high-fiber) at 1.50% of initial body weight for 71 d (1.75% for d 1 to 4). Backfat thickness was recorded ultrasonically at 2-wk intervals, and body weight was recorded at the beginning and end of the 10-wk experiment. Pigs were slaughtered after a 24-h fast, and carcass weight, length and backfat thickness and cross-sectional area of the longissimus muscle were measured. Weights of cecum, heart, liver and kidney and of full and empty stomach and colon and empty small intestine were recorded. Volume and weight of colon and cecum contents were determined. Restriction of digestible energy reduced weight gain to zero or below in pigs fed alfalfa meal compared with 220 g daily in pigs fed the low-fiber diet. Restriction of energy reduced backfat in all three genotypes. Liver, kidney and empty segments of the gastrointestinal tract as a percentage of body weight were increased by high fiber. Obese pigs had smaller longissimus muscle area, more backfat and smaller liver, heart, empty stomach and colon than lean or contemporary pigs, but there were no diet X genotype interactions for any of these traits. Obese pigs consistently had smaller digesta volumes and dry matter weights than the other genotypes. The increased relative organ weights and the associated disproportionate contribution of these organs to body energy expenditure have important implications for effects on basal metabolic rate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of hay substitution on intake and digestibility of forage rape (brassica napus) fed to lambs

Intake and digestibility trials were conducted with sheep to evaluate the effect of adding various levels of a typical fibrous grass forage (neutral detergent fiber, NDF = 68%) to a high quality, low fiber (NDF = 22%) brassica forage. Four forage rape:orchardgrass hay diets (0, 40, 70, 100% rape content on a dry matter basis) were fed to groups of six Polled-Dorset crossbred growing wether lambs (39.6 kg) individually housed in metabolism crates. After a 7-d ad libitum intake period, a 7-d digestibility trial was conducted at 90% of the observed ad libitum intake level. Digestible dry matter intake (DDMI) per unit metabolic body weight increased as rape in the diet was increased from 0 to 70%, with increases in both dry matter intake and dry matter digestibility (DMD). However, DDMI was similar for lambs fed the 70 and 100% rape diets, with DMI decreasing to the same relative extent as DMD increased. Digestibility of the cell wall fraction of the two intermediate diets (40% rape and 70% rape) was lower than predicted from component forage digestibilities. This observation suggests the existence of an associative effect similar to that often reported when forage and concentrates are fed in various ratios. Forage brassicas appear to be utilized in the ruminant in a manner more typically resembling a concentrate than a forage.     

Effects of varying zinc concentrations on quality of alfalfa for lambs

Zinc concentrations in alfalfa hay were varied using a N-Zn liquid fertilizer as a foliar applicant (.34 or .68 kg Zn/ha) or as a soil fertilizer (4.07 kg Zn/ha). Mean concentrations of Zn across five cuttings of alfalfa in 2 yr were 18, 27, 41 and 21 mg Zn/kg DM for control, low foliar, high foliar and soil treatments, respectively. Each treatment was fed in ad libitum amounts to eight crossbred wether lambs (20 to 35 kg) in 6-wk growth and intake trials, followed by 2-wk digestibility and balance trials with individual lambs. For one cutting, hays were also fed in an 81-d trial to four ram lambs (30 to 35 kg) and live weight gain and testicular development were measured. Average daily gain (ADG) and intake over 6 wk differed (P less than .01) with cutting but not with Zn treatment. Average daily gain and testes weight of ram lambs also were not affected by treatment. In the metabolism trials, Zn treatment did not alter (P greater than .05) intake or dry matter digestibility (DMD) of alfalfa, but did influence digestibility of neutral detergent fiber (NDF). Digestible NDF (%) was higher (P less than .05) for high foliar than for low foliar treatments. Apparent absorption and retention of Zn was significantly greater for control than for Zn-treated alfalfas and did not differ with cutting. Mean serum Zn concentrations for control, low and high foliar, and soil treatments were .79, .81, .78 and .75 micrograms Zn/ml, respectively, for all cuttings, with no differences due to treatment.(ABSTRACT TRUNCATED AT 250 WORDS)