Milk fat is a complex mixture of geometric and positional isomers of monounsaturated and polyunsaturated, including short-, long- and branch-chain fatty acids (FAs). There has been partial success to resolve this mixture of FAs using different GC temperature programs, or a combination of GC isothermal and temperature programs. To overcome the problem associated with overlapping isomers prior silver-ion separation was recommended. However, this procedure is time consuming and not practical for routine analysis. In addition, previous methods focused mainly on the trans and cis isomers of 18:1. The present method takes advantage of differences in the relative elution times between different types of FAs. The method involved analyzing each milk fat using the same highly polar 100-m capillary column and GC instrument, and conducting two separations using temperature programs that plateau at 175 and 150 degrees C. The relative shift among the geometric and positional isomers at these two temperature settings was enough to permit identification of most of the trans and cis 16:1, 18:1 and 20:1, the c/t-18:2 and the c/c/t-18:3 isomers found in milk fat. The identity of these FAs was confirmed by prior separation of the total fatty acid methyl esters (FAMEs) of milk fat using Ag(+)-SPE columns, and comparing the fractions to the total milk fat. The Ag(+)-SPE technique was modified to obtain pure saturated, trans- and cis-monounsaturated and diunsaturated FAMEs. By combining the results from these two separate GC analyses, knowing the elution order, it was possible to determine most of the geometric and positional isomers of 16:1, 18:1, 20:1, 18:2 and 18:3 without a prior silver-ion separation. Only few minor FAs could not be resolved, notable the conjugated linoleic acid isomers that still required the complimentary Ag(+)-HPLC separation. The two GC temperature programs have been successfully used to routinely analyze most FA isomers in total milk and beef fats in about 200 min without the use of prior silver-ion separations.
The uniqueness of ruminant milk lipids is based on their high concentration of CLA. Maximal CLA concentrations in milk lipids require optimal conditions of ruminal fermentation and substrate availability, conditions like those present in pasture-fed cows. Our previous work showed that farm management (indoor feeding vs. pasture feeding) markedly influenced the CLA concentration. In this study, the objective was to evaluate the influence of the farm management system as dependent on different locations. Milk samples from different locations (Thuringia and the Alps, representing diverse altitudes) were collected during the summer months and analyzed for FA profile and CLA isomer distribution. The proportion of PUFA and total CLA in milk fat was significantly lower in milk from indoor cows compared with the pasture cows in the Alps. The trans-11 18:1 in milk fat of Alpine cows was elevated, in contrast to lower values for trans-10 18:1. Milk from cows grazing pasture in the Alps was higher in EPA and lower in arachidonic acid than milk from indoor-fed cows. The proportion of cis,trans/trans,cis isomers of CLA was 10 times higher from the indoor cows than from the Alpine cows. In addition to the major isomer cis-9,trans-11, this difference also occurred for the trans-11,cis-13 isomer, which represented more than a fourth of the total CLA present in milk fat. This is the first report showing a special isomer distribution in the milk fat of cows living under very natural conditions. We hypothesize that the CLA isomer trans-11,cis-13 is formed in large quantity as a result of grazing mountain pasture, which is rich in alpha-linolenic acid.
A new method for the spatially resolved measurement of the oxygen saturation of retinal vessels is described. Imaging spectrometry was used for both measurements of transmission and reflectance spectra of whole blood in cuvettes as well as for fundus reflectance spectra. A model was developed for the calculation of the oxygen saturation, valid in the wavelength range between 510 nm and 586 nm, in that the internal reflectance is constant and only the transmitted light depends on layer thickness and hematocrit. Altogether 265 measurements were performed in different number at 30 eyes. In each measurement, the oxygen saturation was simultaneously determined for 193 locations along a line of 1.5 mm at the fundus. The mean oxygen saturation in retinal arteries was (92.2 +/- 4.1)% and (57.9 +/- 9.9)% in retinal veins. The mean retinal arterio-venous difference of the oxygen saturation was (35.1 +/- 9.5)%. The venous oxygen saturation depended on distance from the optic disc. The measured mean of the arterio-venous difference of the oxygen saturation corresponded well to the value of the brain (34%). The utilization of oxygen in the temporal quadrants (inferior: 39.4 +/- 10.4%) is significantly (p = 0.05) higher than in the nasal quadrants (inferior: 31.3 +/- 6.7%).
The aim of this human intervention study was to evaluate the D9-desaturation of trans-11-18 : 1 (trans-vaccenic acid; tVA) to cis-9,trans-11-18 : 2 (c9,t11 conjugated linoleic acid; CLA) and of trans-12-18 : 1 (t12) to cis-9,trans-12-18 : 2 after a short-term (7 d) and a long-term (42 d) supplementation period. The conversion rates of both trans-18 : 1 isomers were estimated by lipid analysis of serum and red blood cell membranes (RBCM). Subjects started with a 2-week adaptation period without supplements. During the 42 d intervention period, the diet of the test group was supplemented with 3 g/d of tVA and 3 g/d of t12. The diet of the control group was supplemented with a control oil. Serum tVA and t12 levels in the test group increased by fivefold and ninefold after 7 d, respectively, and by eight-and 12-fold after 42 d, respectively, when compared with the adaptation period (P#0·002). The serum c9,t11 CLA levels increased by 1·7-and 2·0-fold after 7 d and 42 d, respectively (P#0·001). After 42 d, the test group's RBCM c9,t11 CLA content was elevated by 20 % (P¼ 0·021), whereas in the control group it was decreased by 50 % (P¼ 0·002). The conversion rate of tVA was estimated at 24 % by serum and 19 % by RBCM. No increase in c9,t12-18 : 2 was observed in the serum and RBCM, and thus no conversion of t12 could be determined. In conclusion, the endogenous conversion of dietary tVA to c9,t11 CLA contributes approximately one quarter to the human CLA pool and should be considered when determining the CLA supply.
The objective of this study was to investigate the effects of incremental amounts of Ascophyllum nodosum meal (ANOD) on milk production, milk composition including fatty acids and I, blood metabolites, and nutrient intake and digestibility in early lactation dairy cows fed high-forage diets. Twelve multiparous Jersey cows averaging (mean±standard deviation) 40±21 d in milk and 464±35 kg of body weight and 4 primiparous Jersey cows averaging 75±37 d in milk and 384±17kg of body weight were randomly assigned to treatment sequences in a replicated 4×4 Latin square design. Each period lasted 21 d with 14 d for diet adaptation and 7 d for data and sample collection. Cows were fed a total mixed ration (64:36 forage-to-concentrate ratio) supplemented (as fed) with 0, 57, 113, or 170 g/d of ANOD. Milk yield as well as concentrations and yields of milk components (fat, protein, lactose, milk urea N) were not affected by increasing dietary amounts of ANOD. Concentration (from 178 to 1,370 µg/L) and yield (from 2.8 to 20.6 mg/d) of milk I increased linearly in cows fed incremental amounts of ANOD as a result of the high concentration of I (820 mg/kg of dry matter) in ANOD. Overall, only minor changes were observed in the proportion of milk fatty acids with ANOD supplementation. Quadratic trends were observed for dry matter intake and total-tract digestibilities of organic matter and neutral detergent fiber, whereas negative linear trends were observed for serum concentration of cortisol and crude protein digestibility with ANOD supplementation. Serum concentrations of triiodothyronine and thyroxine were not affected by ANOD supplementation and averaged 1.1 and 48.4 ng/mL, respectively. However, feeding increasing amounts of ANOD linearly reduced the plasma concentration of nonesterified fatty acids (from 164 to 132 mEq/L). Quadratic effects were found for the total-tract digestibility of ADF and urinary output of purine derivatives, suggesting that ANOD supplementation may stimulate growth of ruminal cellulolytic bacteria in a dose-dependent fashion. In general, feeding incremental amounts of ANOD to early lactation dairy cows dramatically increased milk I concentration and output with no effect on animal performance.
Dairy fat and its fatty acids (FAs) have been shown to possess pro-health properties that can support health maintenance and disease prevention. In particular, branched-chain FAs (BCFAs), comprising approximately 2% of dairy fat, have recently been proposed as bioactive molecules contributing to the positive health effects associated with the consumption of full-fat dairy products. This narrative review evaluates human trials assessing the relationship between BCFAs and metabolic risk factors, while potential underlying biological mechanisms of BCFAs are explored through discussion of studies in animals and cell lines. In addition, this review details the biosynthetic pathway of BCFAs as well as the content and composition of BCFAs in common retail dairy products. Research performed with in vitro models demonstrates the potent, structure-specific properties of BCFAs to protect against inflammation, cancers, and metabolic disorders. Yet, human trials assessing the effect of BCFAs on disease risk are surprisingly scarce, and to our knowledge, no research has investigated the specific role of dietary BCFAs. Thus, our review highlights the critical need for scientific inquiry regarding dairy-derived BCFAs, and the influence of this overlooked FA class on human health.
Rumen bacteria form a dynamic, complex, symbiotic relationship with their host, degrading forages to provide volatile fatty acids (VFA) and other substrates as energy to the animal. The objectives were to characterize rumen bacteria in three genetic lines of primiparous dairy cattle, Holstein (HO, n = 7), Jersey (JE, n = 8), and HO × JE crossbreeds (CB, n = 7) across a lactation [3, 93, 183 and 273 days in milk (DIM)] and correlate these factors with VFA, bacterial cell membrane fatty acids (FA), and animal production (i.e. milk yield). This study employed Illumina MiSeq (v. 3) to investigate rumen bacterial communities and gas-liquid chromatography/mass spectroscopy to identify bacterial membrane FA. Lactation stage had a prominent effect on rumen bacterial communities, whereas genetics had a lesser effect on rumen bacteria. The FA composition of bacterial cell membranes was affected by both lactation stage and genetics. Few correlations existed between VFA and bacterial communities; however, moderate correlations occurred between milk yield, protein percentage, fat yield and rumen bacterial communities. Positive correlations were found between branched-chain FA (BCFA) in bacterial cell membranes and bacterial genera. In conclusion, bacterial communities and their FA compositions are more affected by stage of lactation than by genetics of dairy cow.
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