Effect of different fatty acids on the proliferation and cytokine production of dairy cow peripheral blood mononuclear cells

During the transition period, dairy cows often experience negative energy balance, which induces metabolic and immunological disturbances. Our previous work has shown a relationship between the inhibition of immune functions and increased blood nonesterified fatty acid (NEFA) levels. In this study, we evaluated the effect of 11 fatty acids (palmitoleic, myristic, palmitic, stearic, oleic, linoleic, docosahexaenoic, conjugated linoleic, lauric, eicosapentaenoic, and linolenic acids) as well as a mix that represented the NEFA profile observed during the transition period at different concentrations (0, 50, 100, and 250 µM) on proliferation and cytokines secretion of lymphocytes. To assess lymphoproliferation, peripheral blood mononuclear cell (PBMC) from 5 healthy cows (166–189 d in milk) were isolated, stimulated with the mitogenic lectin concanavalin A (ConA), incubated for 72 h with or without fatty acids, and subjected to flow cytometry analysis. Our results showed that all fatty acids, except lauric acid, significantly reduced proliferation of PBMC in a dose-dependent manner. The most detrimental effect was observed with conjugated linoleic and stearic acids, where proliferation of PBMC was already inhibited at the lowest dose (50 µM). For cytokine secretion, we found that levels of IL-4 in culture supernatant of ConA-stimulated PBMC were reduced after a 24-h exposure to the lowest dose (50 µM) of oleic and palmitoleic acids. A dose of 100 µM of eicosapentaenoic acid, NEFA mixture, and myristic acid was necessary to observe a reduction in IL-4 levels. The PBMC also showed a decrease in the secretion of IFN-γ in response to lauric, linolenic, palmitoleic, and stearic acids at 50 µM and myristic acid at 100 µM. Overall, polyunsaturated fatty acids were more potent inhibitors of cytokine secretions than saturated fatty acids. In addition, we detected an inverse relationship between the melting points of fatty acids and their ability to inhibit IL-4 and IFN-γ secretions, as evidenced by greater inhibition with low–melting point fatty acids. Overall, our study confirmed that NEFA have a negative effect on some lymphocyte functions, and that their inhibitory effect on cytokine secretions increases with the degree of unsaturation.     

Cannabinoid-1 receptor activation modulates lipid mobilization and adipogenesis in the adipose tissue of dairy cows

Amplified adipose tissue (AT) lipolysis and suppressed lipogenesis characterize the periparturient period of dairy cows. The intensity of lipolysis recedes with the progression of lactation; however, when lipolysis is excessive and prolonged, disease risk is exacerbated and productivity compromised. Interventions that minimize lipolysis while maintaining adequate supply of energy and enhancing lipogenesis may improve periparturient cows' health and lactation performance. Cannabinoid-1 receptor (CB1R) activation in rodent AT enhances the lipogenic and adipogenic capacity of adipocytes, yet the effects in dairy cow AT remain unknown. Using a synthetic CB1R agonist and an antagonist, we determined the effects of CB1R stimulation on lipolysis, lipogenesis, and adipogenesis in the AT of dairy cows. Adipose tissue explants were collected from healthy, nonlactating and nongestating (NLNG; n = 6) or periparturient (n = 12) cows at 1 wk before parturition and at 2 and 3 wk postpartum (PP1 and PP2, respectively). Explants were treated with the β-adrenergic agonist isoproterenol (1 μM) in the presence of the CB1R agonist arachidonyl-2′-chloroethylamide (ACEA) ± the CB1R antagonist rimonabant (RIM). Lipolysis was quantified based on glycerol release. We found that ACEA reduced lipolysis in NLNG cows; however, it did not exhibit a direct effect on AT lipolysis in periparturient cows. Inhibition of CB1R with RIM in postpartum cow AT did not alter lipolysis. To evaluate adipogenesis and lipogenesis, preadipocytes isolated from NLNG cows' AT were induced to differentiate in the presence or absence of ACEA ± RIM for 4 and 12 d. Live cell imaging, lipid accumulation, and expressions of key adipogenic and lipogenic markers were assessed. Preadipocytes treated with ACEA had higher adipogenesis, whereas ACEA+RIM reduced it. Adipocytes treated with ACEA and RIM for 12 d exhibited enhanced lipogenesis compared with untreated cells (control). Lipid content was reduced in ACEA+RIM but not with RIM alone. Collectively, our results support that lipolysis may be reduced by CB1R stimulation in NLNG cows but not in periparturient cows. In addition, our findings demonstrate that adipogenesis and lipogenesis are enhanced by activation of CB1R in the AT of NLNG dairy cows. In summary, we provide initial evidence which supports that the sensitivity of the AT endocannabinoid system to endocannabinoids, and its ability to modulate AT lipolysis, adipogenesis, and lipogenesis, vary based on dairy cows' lactation stage.     

Functional properties of cream from dairy cows with experimentally altered milk fat composition

Modification of milk fat composition might be a desirable method to alter manufacturing characteristics or produce dairy products low in saturated fat that more closely meet consumer dietary preferences. The aim of this research was to evaluate functional properties of cream obtained from milks with fat composition modified by altering the profile of long-chain fatty acids (FA) absorbed from the intestine. A control and 5 mixtures of long-chain free FA were infused into the abomasum of lactating dairy cows in a 6 × 6 Latin square design with 21-d periods. Treatments were as follows: (1) control (no FA infused), (2) mostly saturated FA (C16:C18 = 0.74), (3) low linoleic palm FA (C16:C18 = 0.73), (4) palm FA (C16:C18 = 0.73), (5) soy FA (C16:C18 = 0.10), and (6) high palmitic soy FA (C16:C18 = 0.73). All treatments included meat solubles and Tween 80 as emulsifiers. Viscosity, overrun, whipping time, foam firmness, and foam stability were evaluated in creams (33% fat). Cream from cows infused with soy FA (treatment 5) had the longest whipping time and lowest overrun, foam stability, viscosity, melting point, firmness, and solid fat content at 5 and 20°C because the fat had the highest unsaturated FA content. Increasing palmitic acid content of soy FA (treatment 6) improved functional variables in cream relative to soy FA alone. Differences among treatments 1 to 4 were less pronounced because of the effect of C18:1 trans in treatments 3 and 4 on milk fat yield and composition. Milk fat from cows infused with palm FA (treatment 4) exhibited comparable or better functionality than control cream. Increased polyunsaturated FA in milk fat resulted in increased amounts of triglyceride (TG) fractions with 28, 30, 38, and 40 carbon numbers, increased oleic acid resulted in increased 50-carbon TG, and higher palmitic and myristic acids resulted in greater 44, 46, and 48 carbon number TG. These TG groups consistently correlated with functional properties of creams from different treatments. Our results indicated that optimal functionality of cream is dependent more on its content of palmitic acid than on unsaturated FA. However, an optimal composition of milk fat for cream functional characteristics might be obtained through nutritional manipulation of diets for dairy cows to deliver an optimal profile of FA.     

Fetuin-A modulates lipid mobilization in bovine adipose tissue by enhancing lipogenic activity of adipocytes

Fetuin-A (FetA) is an adipokine and free fatty acid (FFA) carrier linked to adipose tissue (AT) function in monogastrics and ruminants. In dairy cows, plasma and AT FetA decrease after parturition, coinciding with reduced lipogenesis and increased lipolysis. In monogastrics, FetA enhances lipogenesis, but its role on lipid mobilization of ruminants is unclear. We hypothesized that FetA modulates lipid mobilization in bovine AT by enhancing the lipogenic activity of adipocytes. Our objective was to determine the effects of FetA on lipogenesis and lipolysis in cultured primary adipocytes from dairy cows. Preadipocytes from the tailhead subcutaneous AT depot were induced to differentiate in a 7-d coculture in vitro model. The effects of FetA on lipolytic responses of adipocytes were evaluated after a 2-h β-adrenergic stimulation with 1 µM isoproterenol (ISO) alone or combined with 0.1 mg/mL of FetA (FetA+ISO), and in cells treated with medium alone (CON) or with 0.1 mg/mL of FetA (FetA). Lipogenic responses of adipocytes treated with CON or FetA from d 5 to 7 of differentiation were assessed by fatty acid (FA) uptake quantification and triacylglycerol (TAG) accumulation, and the gene and protein expression of lipogenic markers. Bovine adipocytes abundantly expressed FetA gene and protein and secreted 48 ± 3.5 ng/DNA relative fluorescence units (RFU). Adrenergic stimulation with ISO increased lipolysis compared with CON, as reflected in the release of glycerol (0.12 ± 0.04 vs. 0.04 ± 0.02 nM/DNA RFU) and FFA (15 ± 13 vs. 6.2 ± 2.4 nM/DNA RFU). Lipolysis induced by ISO was attenuated by the addition of FetA (FetA+ISO) as reflected by lower glycerol (0.06 ± 0.04 nM/DNA RFU) and FFA (5.7 ± 2.7 nM/DNA RFU) release compared with ISO alone. Compared with CON, FetA enhanced lipogenic responses as demonstrated by higher FA uptake and increased accumulation of TAG. Exposure to FetA upregulated 1-acylglycerol-3-phosphate acyltransferase-2 (AGPAT2) gene expression and protein content, as well as its activity. Adipocytes exposed to FetA increased the secretion of the metabolite of AGPAT2, phosphatidic acid. In conclusion, FetA attenuates lipolytic responses and enhances lipogenesis in bovine adipocytes. The upregulation of the rate-limiting lipogenic enzyme AGPAT2 by FetA suggests a potential pathway by which this adipokine promotes TAG synthesis in adipocytes. These findings suggest that FetA is a potential target for lipid mobilization modulation in AT of dairy cows.     

Complex formation, thermal properties, and in-vitro digestibility of gelatinized potato starch-fatty acid mixtures

Effects of the type and amount of fatty acid (0-2.0 mmol/g-starch of lauric, myristic, palmitic, stearic, oleic, and linoleic acids) on the complex formation, thermal properties, and in-vitro digestibility of gelatinized potato starch-fatty acid mixtures were investigated. Complex index (CI) evaluated by the reduction in the iodine binding capacity of starch increased with an increase in the amount of fatty acids, and reached a plateau depending on the type of fatty acid. The maximum CI value was higher in the order of lauric (49.9%), linoleic (47.6%), myristic (42.4%), oleic (36.7%), stearic (35.3%), and palmitic acid (30.9%). From the calorimetric study, it was demonstrated that melting temperature of the complexes was higher in the order of stearic (96.7 °C) > lauric, myristic, palmitic, and oleic (89.6-92.1 °C) > linoleic acid (78.3 °C). Melting enthalpy for complexes was roughly related to the CI value (R² = 0.667). From the in-vitro digestibility measurement, it was found that a certain amount of fatty acid reduced the starch content hydrolyzed at a given condition. Among them, 0.50 mmol/g-starch lauric and oleic acid samples showed the largest reduction in the hydrolyzed starch content. This result was related to the extent of complex formation characterized by CI value and its helical order characterized by melting temperature. In addition, there was a possibility of the complex formation between amylose and unsaturated fatty acid during the hydrolysis of gelatinized starch.     

Interrelation of fatty acid composition in adipose tissue, serum, and liver of dairy cows during the development of fatty liver postpartum

We investigated the composition of fatty acids in adipose tissue, serum, and liver of cows that were fed at restricted energy intake or were overfed during the dry period. Overfed cows had higher concentrations of serum nonesterified fatty acids and consequently accumulated greater amounts of triacylglycerols in the liver than did cows that were fed at restricted energy intake. The percentages of the different fatty acids present in adipose tissue were similar for both groups and did not change during sampling intervals. Before parturition, concentrations of the individual fatty acids present in serum were similar between groups. After parturition, concentrations of major fatty acids in serum, including palmitic, stearic, oleic, and linoleic acids significantly increased in both groups and were higher in overfed cows than in cows that were fed at restricted energy intake. The shift of concentrations of the different fatty acids present in the liver--as a result of increased lipolysis-was observed in palmitic, oleic, and linoleic acids but not stearic acid, suggesting that stearic acid is used by the liver (i.e., oxidation) or is considerably secreted through the milk, thereby not increasing in accumulation in the liver. In conclusion, different feeding regimens during the dry period do not influence the composition of fatty acids in adipose tissue. More intensive lipolysis results in increased concentrations of palmitic, stearic, oleic, and linoleic acids in the blood; subsequently, these fatty acids, excluding stearic acid, greatly accumulated in the liver.     

Dietary supplementation with dried olive pomace in dairy cows modifies the composition of fatty acids and the aromatic profile in milk and related cheese

This study aimed to evaluate the effect of dietary integration of dried olive pomace (DOP), a by-product of olive oil separation, on nutritional and aromatic properties of milk and cheese. Twenty dairy cows were divided into 2 groups that were balanced for milk yield, parity, and days in milk. The control group was fed a conventional diet (20 kg of dry matter/head per day), whereas the experimental group (EG) received the conventional diet supplemented with DOP as 10% of dry matter. During the trial, milk yield was recorded and the samples of milk, cheese, total mixed rations, and DOP were collected and analyzed to determine the chemical–nutritional composition and aromatic profile. Atherogenic and thrombogenic indices were calculated on the basis of the fatty acid (FA) profile of milk and cheese. Data were analyzed according to the mixed model for milk yield and chemical composition, including cows nested within treatment as a random effect, whereas the general linear model was used for the analysis of cheese parameters. Differences were assessed by Tukey's test. The EG diet had a lower content of palmitic, stearic, and linoleic acids and a higher level of oleic acid compared with the control. Dietary DOP integration did not affect milk yield and composition with the exception of protein content, which was greater in EG and significantly affected by diet and period. Instead, period was found to be significant for fat and casein in both groups. Dietary supplementation with DOP modified the FA profile of milk and cheese. There was a decrease in short- and medium-chain FA, but significance was achieved only for palmitic acid. The stearic, isomer trans of oleic (in particular vaccenic acid), oleic, and isomer trans of linoleic acids significantly increased. Monounsaturated FA increased in EG milk and cheese and saturated FA were significantly lower, whereas no difference was marked between the groups regarding level of polyunsaturated FA. Supplementation with DOP reduced atherogenic and thrombogenic indices and increased conjugated linoleic acid in both milk and cheese. The free fatty acids, ketones, lactones, esters, and phenylalanine catabolites were increased in raw milk, whereas only leucine metabolism was affected by diet in pasteurized milk cheese at both 1 and 30 d of ripening. The present results pointed out that DOP supplementation may improve the nutritional and nutraceutical properties and modify the aroma of milk and derived cheese.     

EFFECT OF FERMENTATION ON THE FATTY ACID CONTENT AND COMPOSITION OF CASSAVA TUBER MEAL

The effect of fermentation on the fatty acids (FA) content and composition of cassava tuber meal has been investigated. The major FA of the cassava tuber meal (CTM) lipid were oleic and palmitic acids. Other FA found in decreasing order were linoleic, linolenic, palmitoleic, stearic, myristic, pentadecanoic, heptadecanoic and nonadecanoic acids. Fermentation of the CTM resulted in substantial increases in the absolute quantities of the individual FA detected except linolenic acid. However fermentation did not alter the pattern of composition of the FA, but it caused increases in the composition of saturated FA and decreases in certain unsaturated ones. Stearic acid increased in composition by about 92.6% and pentadecanoic by about 50%. A reduction of about 72% of the linolenic acid and 24.2% of the palmitoleic acid composition were obtained in the fermented CTM lipid.     

Component acids of Bauhinia seed oils

Seed oils of Bauhinia monandra and B. purpurea were examined for their component acids by reversed-phase partition column chromatography. the following results (wt. %) were obtained for B. monandra: myristic 1.4, palmitic 15.1, stearic 9.4, arachidic 0.9, behenic 0.9, oleic 11.5, linoleic 60.8 and for B. purpurea: lauric 0.5, myristic 0.5, palmitic 18.5, stearic 17.8, arachidic 1.3, behenic 1.3, oleic 11.1, linoleic 49.0. Epoxy oleic acid reported to be present in the seed oil of B. purpurea was not detected in the sample investigated.     

Total lipid content and fatty acid composition of the seeds of some Vicia L. species

The seed oils of eight Vicia species (Leguminosae) were investigated for their total lipid contents and fatty acid compositions. The seed lipid contents were found to be between 2.30 and 3.91%. The fatty acid compositions of these eight different species were determined by gas chromatography of the methyl esters of their fatty acids. The seed oils of Vicia species contain palmitic and stearic acids as the major component fatty acids, among the saturated acids, with a small amounts of myristic, pentadecanoic, arachidic and behenic acids. The major unsaturated fatty acids found in the seed oils were oleic, linoleic and linolenic acids.     

Hepatic fatty acid composition in periparturient dairy cows with fatty liver induced by intake of a high energy diet in the dry period

The present study compared the hepatic fatty acid composition of cows that were fed a high energy diet during the dry period to induce fatty liver after parturition with that of control cows. Treated cows had higher concentrations of plasma nonesterified fatty acids as a result of greater lipolysis after parturition than did control cows; consequently, the treated cows accumulated greater amounts of triacylglycerols in the liver. Before parturition, treated cows had lower percentages of oleic acid and higher percentages of linoleic acid than did control cows, but percentages of other fatty acids were similar for both groups. After parturition, percentages of each fatty acid were changed substantially, particularly the four major fatty acids (palmitic, stearic, oleic, and linoleic acids). For treated cows, the percentages of palmitic and oleic acids were higher at 0.5 wk after parturition than at 1 wk before parturition; percentages of stearic and linoleic acids decreased. Unlike treated cows, the percentages of both oleic and linoleic acids in the control cows did not change during that time. Moreover, we found that when lipolysis decreased, as indicated by lower plasma nonesterified fatty acid concentrations, the percentages of each fatty acid gradually rebounded toward the concentrations measured before parturition; this observation indicates that the shift in hepatic fatty acid composition is influenced by lipolysis. The increased lipolysis after parturition led to a vast increase in the hepatic triacyglycerol concentration and to a shift in hepatic fatty acid composition.     

Fatty acid composition of raw caprine milk of a native Greek breed during lactation

The fatty acid (FA) and free fatty acid (FFA) profiles of caprine milk from a native Greek breed were investigated during a period of 6 months (January–June). The most abundant FAs were palmitic, oleic, stearic and capric. The short- and medium-chain FAs represented 17.5 and 14.5% of total FA, respectively, while long-chain FAs accounted for the rest (68.0%). From January to April, the major FFAs were acetic, palmitic, stearic and caproic, but afterwards, acetic and propionic were the principal FFAs, followed by stearic, palmitic, oleic and capric.     

Changes in the major free fatty acids in Serra cheese throughout ripening

Changes in the concentrations of the free fatty acids (FFA) containing an even number of carbon atoms in Serra cheese were monitored using a two-way factorial design over a ripening period of 35 days throughout the cheesemaking season. Both time variables had statistically significant effects at the 5% level on the concentrations of FFA. The concentration of stearic acid increased significantly to 351 mg kg⁻¹ (fat basis) only during the first week of ripening. Concentrations of palmitic and oleic acids increased significantly during all stages of ripening and approximately doubled during the 35-day ripening period to reach levels of 425 and 453 mg kg⁻¹, respectively. Concentrations of butyric, caprylic, lauric and myristic acids were approximately constant during the first week and increased significantly thereafter up to 285, 17, 94 and 59 mg kg⁻¹, respectively, at 35 days. The concentrations of caprylic and myristic acids increased five-fold in 35 days, the concentration of butyric acid increased three-fold, and the concentration of lauric acid almost doubled during the same period. The concentrations of caproic, capric, linoleic and linolenic acids increased significantly to 48, 121, 188 and 126 mg kg⁻¹, respectively, only after 21 days of ripening. The concentration of linoleic acid doubled in that time period, while increases in the concentrations of caproic, capric and linolenic acids were much smaller. The concentrations of butyric, caproic and palmitic acids in cheeses manufactured in February were statistically lower than those in cheeses manufactured in May. The principal FFAs throughout ripening were, according to chain length and degree of saturation, butyric (short-chain), capric (medium-chain), palmitic and stearic acids (saturated long-chain), and oleic (unsaturated long-chain). Saturated and unsaturated long-chain FFA were present at the highest concentration at all stages of ripening. It is concluded that lipolysis in Serra cheese proceeds slowly to total FFA concentration of 2167 mg kg⁻¹ (fat basis) by 35 days of ripening.     

Composition of the seed oil of Phalarzs canarienszs

The component fatty acids of the seed oil of Phalaris canuriensis are palmitic, oleic and linoleic with smaller amounts of stearic and linolenic, and a trace of myristic acid. β-Sitosterol was also identified.     

Fatty acid composition of mexican pine nut (Pinus cembroides) oil from three seed coat phenotypes

Fatty acids from oils extracted from three seed coat phenotypes of Mexican stone pines (Pinus cembroides Zuc) growing in the central region of Veracruz State, Mexico, consisted of lauric (0-4.8%), myristic (3.4-9.1%), palmitic (6.4-7.8%), stearic (3.1-5.5%), oleic (36.7-47.2%) and linoleic (32.9-44.5%) acids. The differences between the phenotypes studied were statistically significant in their levels of myristic, oleic and linoleic acids.     

Effect of temperature on fatty acid content in Vicia sativa

The experiment was conducted to evaluate the influence of temperature on fatty acid (FA) concentration of common vetch (Vicia sativa L.). The result suggests that the moderate temperature (23/18?°C) greatly increased the concentration of lipids, palmitic acid, stearic acid, arachidic acid, behenic acid, oleic acid, most notably linoleic acid and linolenic acid, but had limited effects on the concentrations of dodecanoic acid and myristic acid. The level of polyunsaturated FA was higher in 23/18?°C regime as well, and was 3.27-fold higher than that in 30/25?°C regime. Furthermore, the content of unsaturated FAs was 38 and 210?% greater in 23/18?°C regime than in 15/10?°C and 30/25?°C regimes, respectively, and this effect is considered to be associated with the change of membrane fluidity, which could help the plant to fight off temperature stress.     

Prediction of milk fatty acid composition by near infrared reflectance spectroscopy

Near-infrared reflectance spectroscopy (NIRS) (700–2500 nm) was used to predict milk fatty acid (FA) composition. Broad FA variability was ensured by using experimental cow milk derived from different feeding regimes (pasture and preserved forages with or without lipid supplements). Detailed FA composition was analyzed by gas chromatography. Predictive equations (354 samples) were developed for liquid and oven-dried milk samples using modified partial least squares with cross-validation and external validation (114 samples). Coefficient of determination in external validation (R²V) and residual predictive deviation (RPD) were good (R²V ≥ 0.88; RPD ≥ 3.26) for saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), unsaturated fatty acids (UNSAT), trans FA, trans and cis-C18:1, caproic, caprilic, capric, lauric, myristic, palmitic and oleic acids in oven-dried milk, approximate for polyunsaturated fatty acid (PUFA), stearic, vaccenic and rumenic acids (R²V ≤ 0.81; RPD ≤ 3.23) and poor for linoleic, linolenic, total n-6 and n-3 acids. The quantification was more accurate for oven-dried milk, but good results were also obtained for SFA, MUFA, palmitic and oleic acids in liquid milk.     

Analysis of Hiptage madablota seed oil

Hiptage madablota seed kernels yield a pale yellow oil (67·0%) which is rich in ricinoleic acid (70·0%). The seed oil also contains the following acids: caprylic, capric, myristic, palmitic, stearic, oleic, linoleic, arachidic and behenic.     

Changes in plasma triglyceride fatty acids during lactation.

Total blood plasma lipid and plasma triglyceride fatty acids were analyzed at an average of 33, 89, and 139 days of lactation (ranges 20 to 57, 52 to 129, and 122 to 157 days for periods I, II, and III) for 35 Holstein cows in their second or later lactation. Average milk production in the test periods was 35.7, 30.3, and 25.5 kg/day. Lipid analysis was part of a study comparing methionine hydroxy analog or sulfur for lactating cows with effects of treatment and time separated statistically. Cows fed methionine analog had a lower percent palmitate in the triglyceride fatty acid, but there was no other treatment effect. There were significant changes with time in percent total lipid and triglyceride fatty acid. Total lipid averaged 424, 526, and 529 mg/dl. Average percents of measured triglyceride fatty acid and differences with stage of lactation were: myristic, 2.96; palmitic, 27.52, I greater than II or III; palmitoleic, 2.83, I greater than II or III; stearic, 38.58 I greater than II greater than III, oleic, 20.75, I greater than II or III; and linoleic, 4.59.