Potential Biomarkers for Fat from Dairy and Fish and Their Association with Cardiovascular Risk Factors: Cross-sectional Data from the LifeLines Biobank and Cohort Study.
ABSTRACT: Dairy fat intake, reflected by the biomarkers C14:0, C15:0, C17:0, trans-C16:1 (n-7), trans-C18:1 (n-7) and CLA, may have beneficial effects on cardiovascular health. It has, however, been questioned whether this association is genuine, since C15:0 and C17:0 are also biomarkers from fish. We investigated whether the above biomarkers are reliable markers for dairy fat intake in 864 healthy subjects. Subsequently, we explored the association between these biomarkers and cardiovascular risk factors. Intakes of dairy and fish were determined by Food Frequency Questionnaires FFQs. Fatty acids were analyzed in plasma triglycerides (TG) and phospholipids (PL). Median intakes of dairy and fish fat were 12.3 (8.4-17.4) g/day and 1.14 (0.53-1.75) g/day. All fatty acids, except TG C17:0, were associated with dairy fat (std.β range TG: 0.12 for C14:0 till 0.25 for C15:0 and Trans-C18:1 (n-7); and std.β range PL: 0.12 for C17:0 and Trans-C16:1 (n-7) till 0.24 for Trans-C18:1 (n-7) and CLA; p < 0.001). TG C17:0 was associated with fish fat (std.β = 0.08; p = 0.03), whereas PL C17:0 was not. Associations remained after adjustment for fish/dairy fat intake. Strongest inverse associations with biological variables were found with PL C17:0 and Trans-C18:1 (n-7) (Std.βs: waist circumference: -0.18, p < 0.001 and -0.10, p < 0.05; BMI: -0.17, p < 0.001, -0.11, p < 0.01; glucose: -0.10, p <0.01 and -0.08, p <0.05; high sensitive C-reactive protein (hs-CRP): -0.22, p < 0.001 and -0.16, p < 0.01; uric acid: -0.27, p < 0.001 and -0.24, p < 0.001). In conclusion, fatty acid biomarkers, except plasma TG C17:0, were associated with dairy fat intake, independent of fish fat intake. PL C17:0 and trans-C18:1 (n-7) were inversely associated with adiposity, diabetes, inflammation and uric acid.
Project description:Pentadecanoic acid (C15:0) and heptadecanoic acid (C17:0) have been described as dietary biomarkers of dairy-fat consumption, with varying degrees of reliability between studies. It remains unclear how the total amount of dietary fat, representing one of the main confounding factors in these biomarker investigations, affects C15:0 and C17:0 circulating concentrations independent of their relative intake. Additionally, it is not clear how changes in the dietary total-fat affects other fatty acids in circulation. Through two dietary studies with different total-fat levels but maintaining identical fatty acid compositions, we were able to see how the dietary total-fat affects the fatty acids in circulation. We saw that there was a statistically significant, proportionate, and robust decrease in the endogenous C15:0 levels with an increase in dietary total-fat. However, there was no significant change in the circulating C17:0 concentrations as the total-fat increased. To conclude, the dietary total-fat content and fat-type have a very complex influence on the relative compositions of circulating fatty acids, which are independent of the actual dietary fatty acid composition. Knowing how to manipulate circulating C15:0 and C17:0 concentrations is far-reaching in nutritional/pathological research as they highlight a dietary route to attenuate the development of metabolic disease (both by reducing risk and improving prognosis).
Project description:Similar to humans, bottlenose dolphins (Tursiops truncatus) can develop metabolic syndrome and associated high ferritin. While fish and fish-based fatty acids may protect against metabolic syndrome in humans, findings have been inconsistent. To assess potential protective factors against metabolic syndrome related to fish diets, fatty acids were compared between two dolphin populations with higher (n = 30, Group A) and lower (n = 19, Group B) mean insulin (11 ± 12 and 2 ± 5 ?IU/ml, respectively; P < 0.0001) and their dietary fish. In addition to higher insulin, triglycerides, and ferritin, Group A had lower percent serum heptadecanoic acid (C17:0) compared to Group B (0.3 ± 0.1 and 1.3 ± 0.4%, respectively; P < 0.0001). Using multivariate stepwise regression, higher percent serum C17:0, a saturated fat found in dairy fat, rye, and some fish, was an independent predictor of lower insulin in dolphins. Capelin, a common dietary fish for Group A, had no detectable C17:0, while pinfish and mullet, common in Group B's diet, had C17:0 (41 and 67 mg/100g, respectively). When a modified diet adding 25% pinfish and/or mullet was fed to six Group A dolphins over 24 weeks (increasing the average daily dietary C17:0 intake from 400 to 1700 mg), C17:0 serum levels increased, high ferritin decreased, and blood-based metabolic syndrome indices normalized toward reference levels. These effects were not found in four reference dolphins. Further, higher total serum C17:0 was an independent and linear predictor of lower ferritin in dolphins in Group B dolphins. Among off the shelf dairy products tested, butter had the highest C17:0 (423mg/100g); nonfat dairy products had no detectable C17:0. We hypothesize that humans' movement away from diets with potentially beneficial saturated fatty acid C17:0, including whole fat dairy products, could be a contributor to widespread low C17:0 levels, higher ferritin, and metabolic syndrome.
Project description:Dietary odd-chain saturated fatty acids (OCFAs) are present in trace levels in dairy fat and some fish and plants. Higher circulating concentrations of OCFAs, pentadecanoic acid (C15:0) and heptadecanoic acid (C17:0), are associated with lower risks of cardiometabolic diseases, and higher dietary intake of OCFAs is associated with lower mortality. Population-wide circulating OCFA levels, however, have been declining over recent years. Here, we show C15:0 as an active dietary fatty acid that attenuates inflammation, anemia, dyslipidemia, and fibrosis in vivo, potentially by binding to key metabolic regulators and repairing mitochondrial function. This is the first demonstration of C15:0's direct role in attenuating multiple comorbidities using relevant physiological mechanisms at established circulating concentrations. Pairing our findings with evidence that (1) C15:0 is not readily made endogenously, (2) lower C15:0 dietary intake and blood concentrations are associated with higher mortality and a poorer physiological state, and (3) C15:0 has demonstrated activities and efficacy that parallel associated health benefits in humans, we propose C15:0 as a potential essential fatty acid. Further studies are needed to evaluate the potential impact of decades of reduced intake of OCFA-containing foods as contributors to C15:0 deficiencies and susceptibilities to chronic disease.
Project description:BACKGROUND:Odd-numbered chain saturated fatty acids (OCSFA) have been associated with potential health benefits. Although some OCSFA (e.g., C15:0 and C17:0) are found in meats and dairy products, sources and metabolism of C19:0 and C23:0 are relatively unknown, and the influence of non-dietary determinants, including genetic factors, on circulating levels of OCSFA is not established. OBJECTIVE:To elucidate the biological processes that influence circulating levels of OCSFA by investigating associations between genetic variation and OCSFA. DESIGN:We performed a meta-analysis of genome-wide association studies (GWAS) of plasma phospholipid/erythrocyte levels of C15:0, C17:0, C19:0, and C23:0 among 11,494 individuals of European descent. We also investigated relationships between specific single nucleotide polymorphisms (SNPs) in the lactase (LCT) gene, associated with adult-onset lactase intolerance, with circulating levels of dairy-derived OCSFA, and evaluated associations of candidate sphingolipid genes with C23:0 levels. RESULTS:We found no genome-wide significant evidence that common genetic variation is associated with circulating levels of C15:0 or C23:0. In two cohorts with available data, we identified one intronic SNP (rs13361131) in myosin X gene (MYO10) associated with C17:0 level (P = 1.37×10-8), and two intronic SNP (rs12874278 and rs17363566) in deleted in lymphocytic leukemia 1 (DLEU1) region associated with C19:0 level (P = 7.07×10-9). In contrast, when using a candidate-gene approach, we found evidence that three SNPs in LCT (rs11884924, rs16832067, and rs3816088) are associated with circulating C17:0 level (adjusted P = 4×10-2). In addition, nine SNPs in the ceramide synthase 4 (CERS4) region were associated with circulating C23:0 levels (adjusted P<5×10-2). CONCLUSIONS:Our findings suggest that circulating levels of OCSFA may be predominantly influenced by non-genetic factors. SNPs associated with C17:0 level in the LCT gene may reflect genetic influence in dairy consumption or in metabolism of dairy foods. SNPs associated with C23:0 may reflect a role of genetic factors in the synthesis of sphingomyelin.
Project description:We evaluated the between-cow (b-cow) variation and repeatability in omasal and milk fatty acids (FA) related to methane (CH4) emission. The dataset was originated from 9 studies with rumen-cannulated dairy cows conducted using either a switch-back or a Latin square design. Production of CH4 per mole of VFA (Y_CH4VFA) was calculated based on VFA stoichiometry. Experiment, diet within experiment, period within experiment, and cow within experiment were considered as random factors. Empirical models were developed between the variables of interest by univariate and bivariate mixed model regression analysis. The variation associated with diet was higher than the b-cow variation with low repeatability (< 0.25) for milk odd- and branch-chain FA (OBCFA). Similarly, for de novo synthesized milk FA, diet variation was ~ 3-fold greater than the b-cow variation; repeatability for these FA was moderate to high (0.34-0.58). Also, for both cis-9 C18:1 and cis-9 cis-12 cis-15 C18:3 diet variation was more than double the b-cow variation, but repeatability was moderate. Among the de novo milk FA, C4:0 was positively related with stoichiometric Y_CH4VFA, while for OBCFA, anteiso C15:0 and C15:0 were negatively related with it. Notably, when analyzing the relationship between omasal FA and milk FA we observed positive intercept estimates for all the OBCFA, which may indicate endogenous post-ruminal synthesis of these FA, most likely in the mammary gland. For milk iso C13:0, iso C15:0, anteiso C15:0, and C15:0 were positively influenced by omasal proportion of their respective FA and by energy balance. In contrast, the concentration of milk C17:0, iso C18:0, C18:0, cis-11 C18:1, and cis-9 cis-12 cis-15 C18:3 were positively influenced by omasal proportion of their respective FA but negatively related to calculated energy balance. Our findings demonstrate that for most milk FA examined, a larger variation is attributed to diet than b-cow differences with low to moderate repeatability. While some milk FA were positively or negatively related with Y_CH4VFA, there was a pronounced effect of calculated energy balance on these estimates. Additionally, even though OBCFA have been indicated as markers of rumen function, our results suggest that endogenous synthesis of these FA may occur, which therefore, may limit the utilization of milk FA as a proxy for CH4 predictions for cows fed the same diet.
Project description:It is well known that feeding a high concentrate (HC) diet to lactating ruminants likely induces subacute ruminal acidosis (SARA) and leads to a decrease in milk fat production. However, the effects of feeding a HC diet for long periods on milk fatty acids composition and the mechanism behind the decline of milk fat still remains poorly understood. The aim of this study was to investigate the impact of feeding a HC diet to lactating dairy goats on milk fat yield and fatty acids composition with an emphasis on the mechanisms underlying the milk fat depression. Seventeen mid-lactating dairy goats were randomly allocated to three groups. The control treatment was fed a low-concentrate diet (35% concentrate, n = 5, LC) and there were two high-concentrate treatments (65% concentrate, HC), one fed a high concentrate diet for a long period (19 wks, n = 7, HL); one fed a high concentrate diet for a short period of time (4 wk, n = 5, HS). Milk fat production and fatty acids profiles were measured. In order to investigate the mechanisms underlying the changes in milk fat production and composition, the gene expression involved in lipid metabolism and DNA methylation in the mammary gland were also analyzed.Milk production was increased by feeding the HC diet in the HS and HL groups compared with the LC diet (P < 0.01), while the percentage of milk fat was lower in the HL (P < 0.05) but not in the HS group. The total amount of saturated fatty acids (SFA) in the milk was not changed by feeding the HC diet, whereas the levels of unsaturated fatty acids (UFA) and monounsaturated fatty acids (MUFA) were markedly decreased in the HL group compared with the LC group (P < 0.05). Among these fatty acids, the concentrations of C15:0 (P < 0.01), C17:0 (P < 0.01), C17:1 (P < 0.01), C18:1n-9c (P < 0.05), C18:3n-3r (P < 0.01) and C20:0 (P < 0.01) were markedly lower in the HL group, and the concentrations of C20:0 (P < 0.05) and C18:3n-3r (P < 0.01) were lower in the HS group compared with the LC group. However, the concentrations of C18:2n-6c (P < 0.05) and C20:4n-6 (P < 0.05) in the milk fat were higher in the HS group. Real-time PCR results showed that the mRNA expression of the genes involved in milk fat production in the mammary gland was generally decreased in the HL and HS groups compared with the LC group. Among these genes, ACSL1, ACSS1 & 2, ACACA, FAS, SCD, FADS2, and SREBP1 were down-regulated in the mammary gland of the HL group (P < 0.05), and the expressions of ACSS2, ACACA, and FADS2 mRNA were markedly decreased in the HS goats compared with the LC group (P < 0.05). In contrast to the gene expression, the level of DNA methylation in the promoter regions of the ACACA and SCD genes was increased in the HL group compared with the LC group (P < 0.05). The levels of ACSL1 protein expression and FAS enzyme activity were also decreased in the mammary gland of the HL compared with the LC group (P < 0.05).Long-term feeding of a HC diet to lactating goats induced milk fat depression and FAs profile shift with lower MUFAs but higher SFAs. A general down-regulation of the gene expression involved in the milk fat production and a higher DNA methylation in the mammary gland may contribute to the decrease in milk fat production in goats fed a HC diet for long time periods.
Project description:Recent findings have shown an inverse association between circulating C15:0/C17:0 fatty acids with disease risk, therefore, their origin needs to be determined to understanding their role in these pathologies. Through combinations of both animal and human intervention studies, we comprehensively investigated all possible contributions of these fatty acids from the gut-microbiota, the diet, and novel endogenous biosynthesis. Investigations included an intestinal germ-free study and a C15:0/C17:0 diet dose response study. Endogenous production was assessed through: a stearic acid infusion, phytol supplementation, and a Hacl1-/- mouse model. Two human dietary intervention studies were used to translate the results. Finally, a study comparing baseline C15:0/C17:0 with the prognosis of glucose intolerance. We found that circulating C15:0/C17:0 levels were not influenced by the gut-microbiota. The dose response study showed C15:0 had a linear response, however C17:0 was not directly correlated. The phytol supplementation only decreased C17:0. Stearic acid infusion only increased C17:0. Hacl1-/- only decreased C17:0. The glucose intolerance study showed only C17:0 correlated with prognosis. To summarise, circulating C15:0 and C17:0 are independently derived; C15:0 correlates directly with dietary intake, while C17:0 is substantially biosynthesized, therefore, they are not homologous in the aetiology of metabolic disease. Our findings emphasize the importance of the biosynthesis of C17:0 and recognizing its link with metabolic disease.
Project description:The aim of this study was to evaluate the relationship of milk odd- and branched-chain fatty acids (OBCFA) with urinary purine derivates and estimated ruminal microbial crude protein (MCP) synthesis. Forty-four lactating Holstein cows were used in a 4 × 4 Latin square design with 21-day periods comprised of a 13-day adaptation phase to diet followed by an 8-day sampling phase. Differences in estimated MCP yield and milk OBCFA composition were found by feeding total mixed rations containing forage (maize silage, MS; red clover silage, RCS) and concentrates (0.75:0.25) with targeted proportions of RCS to MS of 0.15:0.60, 0.30:0.45, 0.45:0.30, and 0.60:0.15 on a dry matter basis. The MCP was estimated from the total urinary purine derivate (PD) excretion (MCPPD) and intakes of metabolizable energy (MCPME) or digestible organic matter (MCPdOM). The Pearson correlations of individual OBCFA with urinary parameters (uric acid, allantoin, PD and nitrogen) were generally weak (r = -0.37 to 0.55). Yields of individual OBCFA correlated positively with MCPME and MCPdOM (r = 0.21 to 0.55). The prediction of urinary PD concentration was moderate (R2 = 0.64) when including the proportion of iso-C17:0. The prediction of total PD excretion was low (R2 = 0.21) with yields of iso-C15:0, anteiso-C17:0, and iso-C16:0. The prediction of MCPPD was high (R2 = 0.99) when including the iso-C16:0 and cis-9 C17:1 concentrations, while those of MCPME and MCPdOM were low (R2 = 0.37 and 0.36, respectively) when including yields of iso-C15:0, cis-9 C17:1, and iso-C18:0. The correlations and regression analyses demonstrate that the estimated MCP synthesis and urinary PD excretion can be only moderately predicted by yields and concentrations of individual or total OBCFA in cow's milk. However, milk OBCFA can still be seen as a promising, non-invasive method for predicting rumen function and microbial protein supply in dairy cows because MCP flow was not directly measured in this study but instead indirectly estimated probably comprising considerable deviations of the assumed values from the true ones.
Project description:Caenorhabditis elegans FAT-2 has been characterized as fatty acid ?12-desaturase able to desaturate C16 and C18 fatty acids. However, in this report we show that when expressed in yeast cells this enzyme can also catalyze ?15 desaturation. This results in the production of both linoleic acid (?6 C18:2?9,12) and linolenic acid (?3 C18:3?9,12,15) from oleic acid (C18:1?9) substrate, and hexadecadienoic acid (?4 C16:2?9,12) and hexadecatrienoic acid (?1 C16:3?9,12,15) from palmitoleic acid (C16:1?9) substrate. In addition, this enzyme can also produce C14:2?9,12, C15:2?9,12, C17:2?9,12, and C18:4?6,9,12,15 when C14:1?9, C15:1?9, C17:1?9, and C18:3?6,9,12 substrates are available in yeast cells. Mass spectrometry analysis of 2,4-dimethyloxazoline modification of fatty acid methyl esters confirms the positions of all newly formed double bonds. These results indicate that when expressed in yeast the C. elegans ?12-desaturase CeFAT-2 shows a characteristic of a bifunctional ?12/?15-desaturase and has a great deal of elasticity with respect to fatty acid chain length in being able to accept fatty acids ranging from C14 to C18. Interestingly, despite possessing a bifunctional ?12/?15 desaturation activity, phylogenetic analysis suggests that C. elegans ?12-desaturase CeFAT-2 might have arisen independently from other reported dual ?12/?15-desaturases from fungi and protozoa.
Project description:BACKGROUND:Fatty acids (FA) in bovine milk derive through body mobilization, de novo synthesis or from the feed via the blood stream. To be able to digest feedstuff, the cow depends on its rumen microbiome. The relative abundance of the microbes has been shown to differ between cows. To date, there is little information on the impact of the microbiome on the formation of specific milk FA. Therefore, in this study, our aim was to investigate the impact of the rumen bacterial microbiome on milk FA composition. Furthermore, we evaluated the predictive value of the rumen microbiome and the host genetics on the composition of individual FA in milk. RESULTS:Our results show that the proportion of variance explained by the rumen bacteria composition (termed microbiability or [Formula: see text]) was generally smaller than that of the genetic component (heritability), and that rumen bacteria influenced most C15:0, C17:0, C18:2 n-6, C18:3 n-3 and CLA cis-9, trans-11 with estimated [Formula: see text] ranging from 0.26 to 0.42. For C6:0, C8:0, C10:0, C12:0, C16:0, C16:1 cis-9 and C18:1 cis-9, the variance explained by the rumen bacteria component was close to 0. In general, both the rumen microbiome and the host genetics had little value for predicting FA phenotype. Compared to genetic information only, adding rumen bacteria information resulted in a significant improvement of the predictive value for C15:0 from 0.22 to 0.38 (P = 9.50e-07) and C18:3 n-3 from 0 to 0.29 (P = 8.81e-18). CONCLUSIONS:The rumen microbiome has a pronounced influence on the content of odd chain FA and polyunsaturated C18 FA, and to a lesser extent, on the content of the short- and medium-chain FA in the milk of Holstein cattle. The accuracy of prediction of FA phenotypes in milk based on information from either the animal's genotypes or rumen bacteria composition was very low.