Quick changes of milk fatty acids after inclusion or suppression of linseed oil in the diet of goats.

BACKGROUND: Lipid supplementation of ruminant diet is an excellent tool to improve the nutritional quality of dairy fat. The purpose of this research was to monitor in detail the goat milk fatty acid (FA) profile during the first 24 h after linseed oil (LO) supplementation or suppression in the diet. Particular emphasis was placed in the changes of FA with bioactive properties. Milk fat was analysed by gas chromatography from milkings at 0, 1, 3, 6, 12 and 24 h after diet shift. RESULTS: The α-linolenic acid levels increased 12 h after LO incorporation in the diet and decreased 3 h after oil suppression. Most of the milk 10:0 to 16:0 saturated FA decreased 24 h after LO supplementation, whereas oil suppression raised their levels after 6 h. Similarly, raising of mono- and polyunsaturated trans-FA after LO inclusion was delayed in comparison with their decrease after oil suppression. CONCLUSION: This study supports that ruminal bacteria and mammary glands would exhibit a fast responsiveness after the inclusion or suppression of LO in ruminant rations. Milk with an improved FA profile could be collected between 12 h after LO supplementation and the last milking before LO suppression in the diet. © 2018 Society of Chemical Industry.

Probiotic supplementation effects on milk fatty acid profile in ewes.

We hypothesised that probiotic feeding would alter the fatty acid (FA) profile of sheep's milk. Sixteen lactating ewes, kept under the same feeding and management practices, were randomly allocated to receive either a control diet or the same diet supplemented with a commercial multi-strain bacterial probiotic. Milk fat FA contents were monitored fortnightly for eight consecutive weeks from 14 d after lambing. Probiotic supplementation increased the contents of butyric and caproic acids in milk fat and had no negative effects on other relevant FA from the human's health point of view (i.e., no differences in branched chain, vaccenic, rumenic and n-3 FA were observed). Under the conditions assayed in the present work, the contents of milk FA originated from rumen microbial metabolism were scantly altered, which suggests that the rumen conversion pathways of FA were not substantially modified by the probiotics.

Associations between major fatty acids in plant oils fed to dairy goats and C18 isomers in milk fat.

Relationships between fatty acids (FAs) in plant oils included in goat diets and milk fat C18 isomers were determined by Principal Factor Analysis (PFA). The three first principal factors (PF1, PF2 and PF3) accounted for 64.5% of the total variation in milk FAs contents. Fatty acids with a double bond at carbons 13, 14, 15 or 16 had high (>0.6) and positive loadings for PF1, trans-4 to trans-8 C18:1 for PF2, whereas trans-10 C18:1, trans-11 C18:1 and cis-9 trans-11 C18:2 showed high and positive loadings for PF3. Pearson's correlations supported that PF1, PF2 and PF3 were related to α-linolenic, oleic and linoleic acid intakes, respectively. Our results show that the quantitatively main FAs in plant lipids supplemented to dairy ruminants are often the main cause of the observed changes in milk C18 isomer contents. However, sometimes the observed changes are caused, or at least are influenced, by other FAs present in lower quantities in the plant lipids. Thus, using mixtures of plant oils with differently unsaturated main FAs could be a way of tailoring milk fat composition to a pre-designed pattern.

Characterization of cis-9 trans-11 trans-15 C18:3 in milk fat by GC and covalent adduct chemical ionization tandem MS.

Rumen biohydrogenation of dietary alpha-linolenic acid gives rise in ruminants to accumulation of fatty acid intermediates, some of which may be transferred into milk. Rumelenic acid [cis-9 trans-11 cis-15 C18:3 (RLnA)] has recently been characterized, but other C18:3 minor isomers are still unknown. The objective of this work was to identify a new isomer of octatridecenoic acid present in milk fat from ewes fed different sources of alpha-linolenic acid. Structural characterization of this fatty acid was achieved by GC-MS. Analysis of dimethyloxazoline and picolinyl ester derivatives allowed for location of the double bond positions. Covalent adduct chemical ionization tandem mass spectrometry confirmed the positional structure 9-11-15, identical to RLnA, and helped to establish double bond geometry (cis-trans-trans). This new C18:3 isomer could be formed by isomerization of cis-15 bond of RLnA and subsequently converted by hydrogenation to trans-11 trans-15 C18:2, an octadecadienoic acid also detected in this study.

[Trans fatty acids and conjugated linoleic acid in food: origin and biological properties]. / Ácidos grasos trans y ácido linoleico conjugado en alimentos: origen y propiedades biológicas.

INTRODUCTION: Trans fatty acids (TFA) are minor lipid components present in different foods, including ruminant derived products, which have received great attention due to their relationship with cardiovascular disease risk. The origin of TFA in food is mainly related to the industrial hydrogenation processes of unsaturated vegetable oils, but they can also occur naturally in the digestive tract of ruminants by enzymatic biohydrogenation reactions. Both mechanisms generate similar TFA compounds. However, TFA consumption may exert different biological effects depending on the isomeric distribution, which is strongly influenced by the dietary source (i.e., industrial or natural). Industrial hydrogenated vegetable fats are rich in elaidic (trans-9 18:1) and trans-10 18:1 fatty acids, among others. In contrast, vaccenic acid (trans-11 18:1) is the major TFA isomer detected in milk and other ruminant derived products. Vaccenic acid is the physiological precursor of conjugated linoleic acid, a bioactive lipid with beneficial effects on human health. This article provides updated information on the biological effects and potential bioactive properties of TFA considering both, their chemical structure and provenance.

INTRODUCCIÓN: Los ácidos grasos trans (AGT) son componentes lipídicos minoritarios que se encuentran en distintos alimentos, entre ellos, aquellos derivados de animales rumiantes, que han merecido atención por su relación con el riesgo de incidir en enfermedades cardiovasculares. El origen de los AGT en los alimentos se encuentra mayoritariamente en los procesos de hidrogenación industrial de aceites vegetales insaturados y en las reacciones enzimáticas de biohidrogenación que tienen lugar, de forma natural, en el tracto digestivo de los rumiantes. Aunque las moléculas que se generan por ambos mecanismos son similares, la distribución isomérica de los AGT es muy diferente, lo que puede generar diferencias a la hora de evaluar los efectos biológicos derivados de su consumo. Las grasas vegetales hidrogenadas son abundantes en ácido elaídico (trans-9 18:1) y trans-10 18:1 entre otros. En contraste, el ácido vacénico (trans-11 18:1) es el principal AGT presente en la leche y otros productos derivados de rumiantes, siendo además precursor fisiológico del ácido linoleico conjugado, un componente al que se atribuyen numerosos efectos beneficiosos para la salud. En este artículo se actualizan los efectos biológicos y las potenciales propiedades bioactivas de estos ácidos grasos.

Milk and Dairy Product Consumption and Cardiovascular Diseases: An Overview of Systematic Reviews and Meta-Analyses.

Milk and dairy products containing milk fat are major food sources of saturated fatty acids, which have been linked to increased risk of cardiovascular-related clinical outcomes such as cardiovascular disease (CVD), coronary heart disease (CHD), and stroke. Therefore, current recommendations by health authorities advise consumption of low-fat or fat-free milk. Today, these recommendations are seriously questioned by meta-analyses of both prospective cohort studies and randomized controlled trials (RCTs) reporting inconsistent results. The present study includes an overview of systematic reviews and meta-analyses of follow-up studies, an overview of meta-analyses involving RCTs, and an update on meta-analyses of RCTs (2013-2018) aiming to synthesize the evidence regarding the influence of dairy product consumption on the risk of major cardiovascular-related outcomes and how various doses of different dairy products affect the responses, as well as on selected biomarkers of cardiovascular disease risk, i.e., blood pressure and blood lipids. The search strategies for both designs were conducted in the MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews, and Web of Science databases from their inception to April 2018. From the 31 full-text articles retrieved for cohort studies, 17 met the eligibility criteria. The pooled risk ratio estimated for the association between the consumption of different dairy products at different dose-responses and cardiovascular outcomes (CVD, CHD, and stroke) showed a statistically significant negative association with RR values <1, or did not find evidence of significant association. The overview of 12 meta-analyses involving RCTs as well as the updated meta-analyses of RCTs did not result in significant changes on risk biomarkers such as systolic and diastolic blood pressure and total cholesterol and LDL cholesterol. Therefore, the present study states that the consumption of total dairy products, with either regular or low fat content, does not adversely affect the risk of CVD.

Gas chromatography and silver-ion high-performance liquid chromatography analysis of conjugated linoleic acid isomers in free fatty acid form using sulphuric acid in methanol as catalyst.

This study used GC and silver-ion HPLC to examine the effects of temperature and time on methylation of individual and mixtures of conjugated linoleic acid (CLA) isomers in free fatty acid form using sulphuric acid as catalyst. In the conditions tested (temperatures between 20 and 50 degrees C and times between 10 and 60 min) methylation was complete while avoiding isomerization of conjugated dienes and the formation of artefacts that could interfere with chromatographic determinations. An analytical method using solvent extraction of the lipids followed by selective elution of the free fatty acids from aminopropyl bonded phase columns and methylation with H(2)SO(4) in mild conditions was then applied to determine the CLA isomers in free fatty acid form in rumen fluid, and the results were evaluated.

Conjugated linoleic acid in processed cheeses during the manufacturing stages.

Conjugated linoleic acid (CLA) is a naturally occurring micronutrient in milk fat and dairy products consisting of a group of geometric and positional isomers. The purpose of this study was to assess the level and type of CLA isomers found in two commercial processed cheeses (portions and slices) as well as to monitor their evolution during the different manufacturing stages. Total CLA concentrations ranged from 7.5 to 7.9 mg/g of fat, and rumenic acid (cis-9,trans-11 C18:2), the isomer responsible for the biological functions, represented >80% of total CLA. trans-11,cis-13 and trans-11,trans-13 were, with approximately 4% each, the second main CLA isomers. trans-trans isomers accounted for <10% of total CLA. The processing parameters used in this research had negligible effects on the CLA content of processed cheese and did not modify the isomer profile in these dairy products, thereby confirming the stability of rumenic acid during manufacturing.

Changes in the milk and cheese fat composition of ewes fed commercial supplements containing linseed with special reference to the CLA content and isomer composition.

A study was carried out to increase the CLA contents in ewes' milk fat under field conditions by dietary means and to investigate the extent of the changes and consequences for milk processing and cheese quality. During a 3-mon period, ewes' bulk milk samples were collected every week from two different herds. For the first 4 wk the ewes were fed a conventional diet. Then the following 6 wk a supplement enriched in alpha-linolenate (whole linseed) was incorporated into the ovine diet. Finally, in the last 3 wk the feeding was the same as in the first 4 wk. The FA profile in milk fat was monitored by GC, and the distribution of CLA isomers was thoroughly tested by combining GC-MS of 4,4-dimethyloxazoline derivatives (DMOX) with silver ion-HPLC (Ag(+)-HPLC) of FAME. Reconstructed mass spectral profiles of CLA characteristic ions from DMOX were used to identify positional isomers, and Ag(+)-HPLC was used to quantify them. An increase in total CLA in milk fat was observed, and total CLA remained elevated during the weeks of enriched alpha-linolenate feeding. In our experimental conditions there was a linear relationship between trans-vaccenic acid (trans-11-octadecenoic acid; trans-11 18:1) and 9-cis,11-trans CLA in ewes' milk fat. Concerning the CLA isomer profile, increases in the 11,13- and 12,14-18:2 positional isomers were considerable when linseed was included in the diet. Organoleptic characteristics of cheeses made with CLA-enriched milk did not substantially differ from those made with nonsupplemented ewes' milk. CLA total content and isomer profile did not change during ripening.

Oral Absorption and Disposition of alpha-Linolenic, Rumenic and Vaccenic Acids After Administration as a Naturally Enriched Goat Dairy Fat to Rats.

Although there is extensive information describing the positive biological effects of conjugated linoleic acid and its main isomer rumenic acid (RA; C18:2 cis 9, trans 11), and alpha-linolenic acid (ALA) and vaccenic acid (TVA), data about their bioavailability are not available. In this work, we investigated the oral absorption and disposition of these fatty acids in Wistar rats. A naturally enriched goat dairy fat (EDF) was obtained by supplementing ruminant diets with oils or oilseeds rich in polyunsaturated fatty acids (PUFA). The EDF was administered orally (single dose of 3000 mg EDF/kg body weight equivalent to 153 mg TVA/kg body weight, 46 mg RA/kg body weight and 31 mg ALA/kg body weight), and serial blood and liver samples were collected and TVA, RA and ALA concentrations determined by GC/MS. The fatty acids TVA, RA and ALA were rapidly absorbed (t1/2a, 0.36, 0.66 and 0.76 h, respectively, for plasma) and slowly eliminated (t1/2ß, 17.04, 18.40 and 16.52 h, respectively, for plasma). The maximum concentration (C max) was detected in liver > plasma > erythrocyte. Our study shows that when orally administered EDF, its components TVA, RA and ALA were rapidly absorbed and distributed throughout the body by the blood circulation to exert systemic effects.

Mineral balance in milk heated using microwave energy.

Milk heated to 75 and 85 degrees C in a water bath or in a microwave oven was assayed for changes in salt partitioning after cooling to room temperature. To properly to assess differences and draw valid comparisons, the two heating methods used in the experiment were applied to samples for identical exposure times, and the samples were heated to attain the same final temperatures. Although the soluble Ca and P(i) contents were lower in the heated milk samples, no significant differences in salt partitioning were found between microwave and conventional heating. Ionic calcium levels in the milk samples pasteurized using microwave energy were very close to the levels in the samples heated in a conventional water bath (approximately 90% of the level in the untreated milk samples). The microwave heating-induced changes were completely reversed after storage at 20 degrees C for 24 h. The coagulation properties of the heated milk samples were also examined, and the coagulation time was longer and the curd formation rate slower in the microwave-heated milk than in the raw milk. Still, the experimental results demonstrated that microwave heating was no more detrimental to the milk than conventional heating and could thus be used for pasteurization purposes.

A 4-week repeated oral dose toxicity study of dairy fat naturally enriched in vaccenic, rumenic and α-linolenic acids in rats.

Few studies have focused on the toxicological risks of dairy fat intake. A standard dairy fat (SDF) with a 70% SFA content and a naturally enriched dairy fat (EDF) in vaccenic, rumenic and α-linolenic acids and low in SFA (54%) have been examined in a 4-week repeated dose oral toxicity study as a daily dose of 2000 mg/kg bw by gavage in rats. Comparisons were established with a third group of rats (control) which did not receive fat administration. Both fats were well tolerated, and no adverse events or mortality were observed during the treatment nor after a 2-week observation period. EDF and SDF did not cause significant differences with respect to a control group in body weight gain, food consumption, clinical observations, organ weight ratios, histopathological findings and most of the hematological and biochemical parameters including total cholesterol and cholesterol fractions in plasma. In rats treated with SDF, a significant increase of triglycerides was observed as compared to the control group. By contrast, in rats treated with EDF, a significant decrease in triglycerides was detected. EDF orally administered to rats was safe, and no treatment-related toxicity was detected. The results also suggest that EDF could protect against the increase of triglyceride concentrations in plasma.

Comparison of the fatty acid profiles in cheeses from ewes fed diets supplemented with different plant oils.

The purpose of this work was to obtain a cheese from ewes milk with a healthier fatty acid (FA) profile. To achieve our aim, 48 ewes (12 per treatment) were fed diets supplemented with 3% of plant oils: palm (used as control), olive (OO), soybean (SO), and linseed (LO). Milk samples from each treatment were collected to manufacture cheeses. The cheesemaking process did not modify the dairy fat FA profile, but OO, SO, and LO did reduce the C12:0 + C14:0 + C16:0 content in dairy fat, thus decreasing the atherogenic index value in the cheeses. Percentages of cis-9 trans-11 C18:2 in cheeses ranged from the 0.43 control value to 0.92, 1.64, and 2.71 with OO, LO, and SO respectively, following the same pattern as trans-11 C18:1. In contrast, trans-10 C18:1 levels were always below 1%. The lowest n-6/n-3 ratio obtained with LO (1.43) suggests that such lipid supplementation would be the most effective nutritional strategy for improving cheese FA profiles.

Acute oral safety study of dairy fat rich in trans-10 C18:1 versus vaccenic plus conjugated linoleic acid in rats.

The acute oral toxicity of a trans-10 C18:1-rich milk fat (T10, 20% of total FA), and a trans-11 C18:1+cis-9 trans-11 C18:2-rich milk fat (T11-CLA, 14% and 4.8% of total FA, respectively) was studied in rats receiving a single oral dose of 2000 mg/kg body weight (BW). T10 and T11-CLA milk fats were well tolerated; no adverse effects or mortality were observed during the 2-week observation period. Two weeks following a single oral dose of 2000 mg/kg BW of T10 and T11-CLA milk fats there were no changes in haematological and serum chemistry parameters (excepting plasma lipid) organ weights, gross pathology or histopathology. In rats treated with T10 milk fat a significant increase of triglycerides was observed. In contrast, in rats treated with T11-CLA milk fat significantly decreased triglycerides were detected. It was concluded that dairy fats rich in T10 and T11-CLA have a low order of acute toxicity, the oral lethal dose (DL50) for male and female rats are in excess of 2000 mg/kg BW. Our results suggest that the T10 milk fat treatment tended to increase triglycerides concentrations, whereas the T11-CLA milk fat treatment tended to reduce it.

Effect of diet supplementation with sunflower oil on milk production, fatty acid profile and ruminal fermentation in lactating dairy ewes.

The aim of this research was to enhance the nutritional quality of ewe milk fat by increasing potentially healthy fatty acids (FA) through diet supplementation with unprotected oil rich in linoleic acid, and without detrimental effects on animal performance. Twenty-four ewes were assigned to two high concentrate diets, control or supplemented with 6% sunflower oil (SO), for 4 weeks. No differences between treatments were found in milk production and dry matter intake. Although the SO diet increased milk fat percentage and tended to reduce milk protein concentration, it did not affect milk fat, protein or total solid yield. Most of the modifications in milk FA composition were addressed toward a potentially healthier profile: a decrease in C12:0 to C16:0 and a remarkable increase in the contents of cis-9 trans-11 C18:2 (from 0.94 to 3.60 g/100 g total FA) and trans-11 C18:1 (from 2.23 to 8.61 g/100 g total FA). Furthermore, the levels reached were maintained throughout the period monitored. However, the SO diet increased other trans C18:1 isomer percentages, too. The lack of differences between treatments in the in vitro ruminal fermentation parameters, studied with batch cultures of rumen microorganisms, would indicate no negative effects on ruminal fermentation.

Authenticity assessment of dairy products.

The authenticity of dairy products has become a focal point, attracting the attention of scientists, producers, consumers, and policymakers. Among many others, some of the practices not allowed in milk and milk products are the substitution of part of the fat or proteins, admixtures of milk of different species, additions of low-cost dairy products (mainly whey derivatives), or mislabeling of products protected by denomination of origin. A range of analytical methods to detect frauds have been developed, modified, and continually reassessed to be one step ahead of manufacturers who pursue these illegal activities. Traditional procedures to assess the authenticity of dairy products include chromatographic, electrophoretic, and immunoenzymatic methods. New approaches such as capillary electrophoresis, polymerase chain reaction, and isotope ratio mass spectrometry have also emerged alongside the latest developments in the former procedures. This work intends to provide an updated and extensive overview since 1991 on the principal applications of all these techniques together with their advantages and disadvantages for detecting the authenticity of dairy products. The scope and limits of different tools are also discussed.

Conjugated linoleic acid in ewe milk fat.

Ewe milk fat from five different herds was studied to determine the content of conjugated linoleic acid (CLA) isomers. Research was carried out by combining gas chromatography-mass spectrometry (GC-MS) of fatty acid methyl esters (FAME) and 4,4-dimethyloxazolyne derivatives (DMOX) with silver ion-high performance liquid chromatography (Ag+-HPLC). Reconstructed mass spectral profiles of CLA characteristic ions from DMOX were used to identify positional isomers and Ag+-HPLC to quantify them. Total CLA content varied from 0.57 to 0.97 g/100 g of total fatty acids. FAME and DMOX were separated into a complex mixture of minor isomers and major rumenic acid (9-cis 11-trans C18:2) by GC-MS using a 100-m polar capillary column. Rumenic acid would represent more than 75% of total CLA. 11-trans 13-trans, 11-13 cis/trans plus trans/cis and 7-9 cis/trans plus trans/cis were the main CLA isomers after rumenic acid. Minor amounts of 8-10 and 10-12 C18:2 isomers were also found. Although most of the isomers were present in each herd's milk, differences in content were observed for some CLA species.