Fatty Acid Methyl Ester Analysis Research Articles

Identification of the major yeasts isolated from high moisture corn and corn silages in the United States using genetic and biochemical methods

The objective of this study was to identify species of yeasts in samples of high moisture corn (HMC) and corn silage (CS) collected from farms throughout the United States. Samples were plated and colonies were isolated for identification using DNA analysis. Randomly selected colonies were also identified by fatty acid methyl esters (FAME) and by physiological substrate profiling (ID 32C). For CS, Candida ethanolica, Saccharomyces bulderi, Pichia anomala, Kazachstania unispora, and Saccharomyces cerevisiae were the predominant yeasts. Pichia anomala, Issatchenkia orientalis, S. cerevisiae, and Pichia fermentans were the prevalent species in HMC. The 3 identification methods were in agreement at the species level for 16.6% of the isolates and showed no agreement for 25.7%. Agreement in species identification between ID 32C and DNA analysis, FAME and ID 32C, and FAME and DNA analysis was 41.1, 14.4, and 2.2%, respectively. Pichia anomala and I. orientalis were able to grow on lactic acid, whereas S. cerevisiae metabolized sugars (galactose, sucrose, and glucose) but failed to use lactic acid. The yeast diversity in CS and HMC varied due to type of feed and location. Differences in species assignments were seen among methods, but identification using substrate profiling generally corresponded with that based on DNA analysis. These findings provide information about the species that may be expected in silages, and this knowledge may lead to interventions that control unwanted yeasts.

Non-conjugated cis/trans 18:2 in Beef Fat are Mainly Δ-9 Desaturation Products of trans-18:1 Isomers.

Human liver cells (HepG2) were cultured with individual trans (t) 18:1 including t6-, t12-, t13-, t14-, t15- and t16-18:1, and retention times of their Δ-9 desaturation products were determined using 100-m biscyanopropyl-polysiloxane and SLB-IL111 columns. Corresponding peaks were found in beef adipose tissues known to have different delta-9 desaturase activities. Further lines of evidence indicating the presence of Δ-9 desaturation products of t-18:1 isomers in beef fat were developed by analysis of fatty acid methyl esters (FAME) fractionated using Ag+-TLC, and by GC/MS. Some of the Δ-9 desaturation products of t-18:1 have been previously identified in ruminant fat (c9, t12- and c9, t13-18:2). Some of the Δ-9 desaturation products of t-18:1 (c9, t14- and c9, t15-18:2) have been previously tentatively identified as different fatty acids, and for the first time we provide evidence of the presence of c9, t16-18:2, and where t6, c9-18:2 may elute during analysis of FAME from beef fat.

Effect of flue gas CO2 on the growth, carbohydrate and fatty acid composition of a green microalga Scenedesmus obliquus for biofuel production

ABSTRACT Effect of various flue gas CO2 concentrations (5%, 10% and 14.1%) on growth rate and biochemical properties of a green microalga Scenedesmus obliquus was investigated. S. obliquus showed the highest biomass production and growth rate (0.36 g L−1 and μ max = 1.00 day−1), total inorganic carbon removal (35.8 mg L−1), lipid productivity (9.9 mg L−1 day) and carbohydrate productivity (10.3 mg L−1 day) with 14.1% CO2 after 8 days of cultivation. Fatty acid methyl ester analysis revealed that the palmitic and oleic acid contents were increased up to 5% and 7% with 14.1% CO2, respectively. Application of flue gas CO2 enhanced the growth along with lipid and carbohydrate productivity of S. obliquus, which can be exploited for reducing the CO2 concentration.

Extraction and Analysis of Microbial Phospholipid Fatty Acids in Soils.

Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about the overall structure of terrestrial microbial communities. PLFA profiling has been extensively used in a range of ecosystems as a biological index of overall soil quality, and as a quantitative indicator of soil response to land management and other environmental stressors.The standard method presented here outlines four key steps: 1. lipid extraction from soil samples with a single-phase chloroform mixture, 2. fractionation using solid phase extraction columns to isolate phospholipids from other extracted lipids, 3. methanolysis of phospholipids to produce fatty acid methyl esters (FAMEs), and 4. FAME analysis by capillary gas chromatography using a flame ionization detector (GC-FID). Two standards are used, including 1,2-dinonadecanoyl-sn-glycero-3-phosphocholine (PC(19:0/19:0)) to assess the overall recovery of the extraction method, and methyl decanoate (MeC10:0) as an internal standard (ISTD) for the GC analysis.

Extraction and Analysis of Microbial Phospholipid Fatty Acids in Soils

Phospholipid fatty acids (PLFAs) are key components of microbial cell membranes. The analysis of PLFAs extracted from soils can provide information about the overall structure of terrestrial microbial communities. PLFA profiling has been extensively used in a range of ecosystems as a biological index of overall soil quality, and as a quantitative indicator of soil response to land management and other environmental stressors. The standard method presented here outlines four key steps: 1. lipid extraction from soil samples with a single-phase chloroform mixture, 2. fractionation using solid phase extraction columns to isolate phospholipids from other extracted lipids, 3. methanolysis of phospholipids to produce fatty acid methyl esters (FAMEs), and 4. FAME analysis by capillary gas chromatography using a flame ionization detector (GC-FID). Two standards are used, including 1,2-dinonadecanoyl-sn-glycero-3-phosphocholine (PC(19:0/19:0)) to assess the overall recovery of the extraction method, and methyl decanoate (MeC10:0) as an internal standard (ISTD) for the GC analysis.

Biodiesel and poly-unsaturated fatty acids production from algae and crop plants - a rapid and comprehensive workflow for lipid analysis.

Fatty acid methyl ester analysis (FAME) by gas chromatography coupled to mass spectrometry (GC-MS) is a widely used technique in biodiesel/bioproduct (e.g. poly-unsaturated fatty acids, PUFA) research but typically does not allow distinguishing between bound and free fatty acids. To understand and optimize biosynthetic pathways, however, the origin of the fatty acid is an important information. Furthermore the annotation of PUFAs is compromised in classical GC-EI-MS because the precursor molecular ion is missing. In the present protocol an alkaline methyl esterification step with TMS derivatization enabling the simultaneous analysis of bound and free fatty acids but also further lipids such as sterols in one GC-MS chromatogram is combined. This protocol is applied to different lipid extracts from single cell algae to higher plants: Chlorella vulgaris, Chlamydomonas reinhardtii, Coffea arabica, Pisum sativum and Cuscuta japonica. Further, field ionization (GC-FI-MS) is introduced for a better annotation of fatty acids and exact determination of the number of double bonds in PUFAs. The proposed workflow provides a convenient strategy to analyze algae and other plant crop systems with respect to their capacity for third generation biodiesel and high-quality bioproducts for nutrition such as PUFAs.

Enterococcus saigonensis sp. nov., isolated from retail chicken meat and liver.

Two Gram-stain-positive strains, VE80T and VE116, which were resistant to vancomycin, were isolated from retail chicken meat and liver in Ho Chi Minh, Vietnam, respectively. These strains were characterized by sequence analyses of 16S rRNA, RNA polymerase α-subunit (rpoA), ATP synthase α-subunit (atpA), and phenylalanyl-tRNA synthase α-subunit (pheS) genes, determination of DNA G+C content, cellular fatty acid methyl ester analysis, DNA-DNA hybridization, and conventional morphological and biochemical tests. Strains VE80T and VE116 had 99.6 % 16S rRNA gene sequence similarity with Enterococcus canintestini LMG 13590T, and 99.1 % 16S rRNA gene sequence similarity with Enterococcus dispar ATCC 51266T. However, the two isolates could be clearly differentiated from these reference strains by the low sequence similarities (86.1-86.8 %) of the atpA gene, low DNA-DNA relatedness (<22.8 %), and differences in the production of acid from melezitose and methyl α-d-glucoside. Based on the results obtained in the present study, these two isolates are considered to represent a novel species of the genus Enterococcus, for which the name Enterococcus saigonensis sp. nov., is proposed. The type strain is VE80T (=JCM 31193T=CCUG 68827T).

Evaluation of Waste Loquat Kernels as Substrate for Lipid Production by Rhodotorula glutinis SO28

This study was performed to produce lipids from the isolated oleaginous yeast Rhodotorula glutinis SO28 using loquat kernel extract (LKE) as substrate. LKE was prepared using acid hydrolysis and alkaline neutralization steps. Lipid production was performed in shaking flaks culture. Even if LKE was used as a sole source of nutritional substances, it could support cell growth and lipid synthesis in the yeast. Additional carbon, nitrogen and phosphorus sources were found to significantly alter the lipid accumulation potential of the yeast. Optimal concentrations of additional carbon (glucose) and nitrogen (ammonium sulphate) sources for lipid accumulation were determined as 15 and 0.5 g/L, respectively. On the other hand, all the concentrations of additional phosphorus source were found to significantly reduce the lipid accumulation. Optimal incubation time was determined as 132 h. Under the optimized culture conditions, the lipid concentration and lipid content of the yeast were determined as 7.82 g/L and 62 %, respectively. Fatty acid methyl ester analysis exhibited that this yeast strain could produce high proportions of C16:0 and C18 fatty acids, which are ideal for biodiesel production. This is the first report on the use of waste loquat kernels as substrate for microbial lipid production.

Microalga, Acutodesmus obliquus KGE 30 as a potential candidate for CO2 mitigation and biodiesel production.

In this study, the effect of flue gas CO2 on growth, lipid production, and fatty acid composition of a green microalga Acutodesmus obliquus KGE 30 was investigated. The highest growth rate (0.46gL(-1) and μmax = 1.09day(-1)), total inorganic carbon removal (95.9mgL(-1)), and lipid productivity (20.1mgL(-1) day L(-1)) was obtained at 14.1% CO2 after 4days of cultivation. In a semicontinuous batch reactor, the highest biomass production (1.19gL(-1)) was achieved after 12days with continuous injection of flue gas CO2. Compared with synthetic CO2, fatty acid methyl ester analysis showed that the amount of unsaturated fatty acid increased by 19.2% with 14.1% flue gas CO2. The application of flue gas CO2 improved biomass production and lipid productivity in A. obliquus. The current investigation demonstrated that the use of flue gas CO2 could reduce the cost of microalgae biomass production for better biofuel generation.

Comparative studies of immobilized lipase- and acid-catalyzed fatty acid methyl ester synthesis for seed lipid analysis.

Fatty acids are one of the most important nutrients in food. Acid- or base-catalyzed transesterification methods are commonly used for the analysis of fatty acids. However, several drawbacks were reported for these methods, including the isomerization and degradation of fatty acids. Lipase-catalyzed reactions are usually undertaken at mild conditions, preventing such problems. In this study, commercial resin-bound lipase from Candida antartica was tested for possible application in fatty acid methyl ester analysis. Experimental parameters, including temperature, reaction time, and re-cycling were evaluated. The optimized condition was (5-10 mg lipid, 0.5 mL of MeOH, and 50 mg Novozyme 435 in 2 mL toluene, 80°C for 1 h). In optimized condition, the lipase-catalyzed methods yielded similar results with the chemical method. In overall, lipase-catalyzed transesterification can be a useful alternative to acid-catalyzed methods for fatty acid analysis.

Effect of cryogenic grinding on volatile and fatty oil constituents of cumin (Cuminum cyminum L.) genotypes.

Effect of cryogenic grinding on recovery of volatile oil, fatty oil percentage and their constituents in two cumin (Cuminum cyminum L.) genotypes have been analyzed. Cryogenic grinding not only retains the volatiles but enhanced the recovery by 33.9% in GC 4 and 43.5% in RZ 209. A significant increase (29.9%) over normal grinding in oil percentage was also observed in genotype RZ 209. This increase was, however, less (15.4%) in genotype GC 4. Nineteen major compounds were identified in the essential oil of both genotypes. The two grinding techniques had significant effects on dependent variables, viz., volatile oil and monoterpenes. Cuminaldehyde was the main constituent in both genotypes, content of which increased from 48.2 to 56.1% in GC 4 on cryo grinding. Content of terpines were found to decrease in cryo ground samples of GC 4 and either decrease or no change was found in RZ 209. Organoleptic test showed more pleasant aroma in cryo ground seeds of both the genotypes. Significant increase was also reported in fatty oil yield due to cryogenic grinding. Fatty acid methyl ester (FAME) analysis showed oleic acid as major FAME content of which increased from 88.1 to 94.9% in RZ 209 and from 88.2 to 90.1% in GC 4 on cryogenic grinding. Other prominent FAME were palmitic, palmitoleic and stearic acid. Results indicated commercial potential of cryogenic grinding technology for cumin in general and spices in particular for better retention of flavour and quality in spices.

Wild almond (Prunus scoparia L.) as potential oilseed resource for the future: Studies on the variability of its oil content and composition

Wild almond genetic resources have still not received considerable attention for oil chemical compositions and uses. The aim of this study was to assess the levels of variation in oil content and fatty acid composition in forty Iranian accessions of Prunus scoparia L. (Spach) to identify genotypes with desirable traits in terms of oil quantity, quality and industrial utilization. Oil parameters and indices were measured, and fatty acid methyl ester analysis was carried out by gas liquid chromatography. Oleic and linoleic fatty acids showed high variability among accessions, ranging from 232.4 to 359.6g/kg oil and from 190.7 to 348.8g/kg oil, respectively. Total unsaturated fatty acid fraction was higher than total saturated fatty acid. The ranges of saponification number (199.2–202.1), iodine value (104.8–125.7kg I2/kg) and cetane number (43.8–48.8), confirmed that the oils have industrial potentialities. Results could contribute to select wild almond genotypes as genetic sources for oil production.

An Overview of Different Techniques on the Microbial Community Structure, and Functional Diversity of Plant Growth Promoting Bacteria

Soil is a dynamic biological system, in which it is difficult to determine the composition of microbial communities. Knowledge of microbial diversity and function in soils are limited because of the taxonomic and methodological limitations associated with studying the organisms. In this review, approaches to measure microbial diversity in soil were discussed. Research on soil microbes can be categorized as structural diversity, functional diversity and genetic diversity studies, and these include cultivation based and cultivation independent methods. Cultivation independent technique to evaluate soil structural diversity include different techniques such as Phospholipid Fatty Acids (PLFA) and Fatty Acid Methyl Ester (FAME) analysis. Carbon source utilization pattern of soil microorganisms by Community Level Physiological Profiling (CLPP), catabolic responses by Substrate Induced Respiration technique (SIR) and soil microbial enzyme activities are discussed. Genetic diversity of soil microorganisms using molecular techniques such as 16S rDNA analysis Denaturing Gradient Gel Electrophoresis (DGGE) / Temperature Gradient Gel Electrophoresis (TGGE), Terminal Restriction Fragment Length Polymorphism (T-RFLP), Single Strand Conformation Polymorphism (SSCP), Restriction Fragment Length Polymorphism (RFLP) / Amplified Ribosomal DNA Restriction Analysis (ARDRA) and Ribosomal Intergenic Spacer Analysis (RISA) are also discussed. The chapter ends with a final conclusion on the advantages and disadvantages of different techniques and advances in molecular techniques to study the soil microbial diversity.

Optimization of lipid extraction from Salvinia molesta for biodiesel production using RSM and its FAME analysis.

The higher areal productivity and lipid content of microalgae and aquatic weed makes them the best alternative feedstocks for biodiesel production. Hence, an efficient and economic method of extracting lipid or oil from aquatic weed, Salvinia molesta is an important step towards biodiesel production. Since Salvinia molesta is an unexplored feedstock, its total lipid content was first measured as 16% using Bligh and Dyer's method which was quite sufficient for further investigation. For extracting more amount of lipid from Salvinia molesta, methanol: chloroform in the ratio 2:1 v/v was identified as the most suitable solvent system using Soxhlet apparatus. Based on the literature and the preliminary experimentations, parameters such as solvent to biomass ratio, temperature, and time were identified as significant for lipid extraction. These parameters were then optimized using response surface methodology with central composite design, where experiments were performed using twenty combinations of these extraction parameters with Minitab-17 software. A lipid yield of 92.4% from Salvinia molesta was obtained with Soxhlet apparatus using methanol and chloroform (2:1 v/v) as solvent system, at the optimized conditions of temperature (85°C), solvent to biomass ratio (20:1), and time (137min), whereas a predicted lipid yield of 93.5% with regression model. Fatty acid methyl ester (FAME) analysis of S. molesta lipid using gas chromatograph mass spectroscopy (GCMS) with flame ionization detector showed that fatty acids such as C16:0, C16:1, C18:1, and C18:2 contributed more than 9% weight of total fatty acids. FAME consisted of 56.32, 28.08, and 15.59% weight of monounsaturated, saturated, and polyunsaturated fatty acids, respectively. Higher cetane number and superior oxidation stability of S. molesta FAME could be attributed to its higher monounsaturated content and lower polyunsaturated content as compared to biodiesels produced from C. vulgaris, Sunflower, and Jatropha.

The isolation and identification of new microalgal strains producing oil and carotenoid simultaneously with biofuel potential

Taxonomy and phylogeny of twenty two microalgal isolates were examined using both universal and newly designed molecular primers. Among the isolates, Scenedesmus bijugus, Coelastrella sp., Auxenochlorella protothecoides, and Chlorella sp. were particularly promising in terms of producing lipids as measured by fatty acid methyl esters (FAME) analysis and significant concentration of carotenoids. A comparative experiment showed that S. bijugus and Chlorella sp. were the most promising candidates (L−1d−1, with biomass) 174.77±6.75, 169.81±5.22mg, lipids 40.14±3.31, 39.72±3.89mg, lutein 0.47, 0.36mg, and astaxanthin 0.27, 0.18mg respectively. The fatty acids produced by these microalgal isolates were mainly palmitic, stearic, oleic, linoleic, and linolenic acid. The freshwater microalgal isolate S. bijugus be the most suitable isolate for producing biodiesel and carotenoids, due to high productivity of biomass, lipids, metabolites, and its suitable fatty acid profile.

Large-scale gene expression profiling reveals physiological response to deletion of chaperone dnaKJ in Escherichia coli

Chaperone DnaK and its co-chaperone DnaJ plays various essential roles such as in assisting in the folding of nascent peptides, preventing protein aggregation and maintaining cellular protein homeostasis. Global transcriptional changes in vivo associated with deletion of dnaKJ were monitored using DNA microarray to elucidate the role of DnaKJ at the transcriptional level. Microarray profiling and bioinformatics analysis revealed that a few chaperone and protease genes, stress-related genes and genes involved in the tricarboxylic acid cycle and oxidative phosphorylation were up-regulated, whereas various transporter genes, pentose phosphate pathway and transcriptional regulation related genes were down-regulated. This study is the first to systematically analyze the alterations at the transcriptional level in vivo in deletion of dnaKJ. Fatty acid methyl esters analysis indicated that the amount of unsaturated fatty acid sharply increased and subcellular location prediction analysis showed a marked decrease in transcription of inner-membrane protein genes, which might have triggered the development of aberrant cell shape and susceptibility for some antibiotics in the ΔdnaKJ strain.

Analysis of optimal conditions for biodiesel production from Jatropha oil in supercritical methanol: Quantification of thermal decomposition degree and analysis of FAMEs

Recent years have seen great efforts made to optimise the production of biodiesel as an alternative to fossil fuel. This study looks at the optimal conditions for producing second generation biodiesel in supercritical methanol from Jatropha oil. Triglyceride conversion and the yield of monoglycerides, diglycerides and fatty acid methyl esters (FAMEs) are analysed for different ranges of temperature and time. The maximum conversion of triglycerides (100wt%) and maximum yield of FAMEs (99.5mol%) were achieved at 325°C with a reaction time of 90min. The thermal decomposition of the biodiesel produced was greatest (24.16%) at 350°C and 90min. Quantification of individual methyl esters showed that the fatty acid chains affected by temperature were those of oleic and linoleic acids. Finally, a lineal regression model was applied to predict the effect of temperature on the biodiesel produced.

Apibacter mensalis sp. nov.: a rare member of the bumblebee gut microbiota.

Isolates LMG 28357T (=R-53146T) and LMG 28623 were obtained from gut samples of Bombus lapidarius bumblebees caught in Ghent, Belgium. They had identical 16S rRNA gene sequences which were 95.7 % identical to that of Apibacter adventoris wkB301T, a member of the family Flavobacteriaceae. Both isolates had highly similar matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS and randomly amplified polymorphic DNA (RAPD) profiles. A draft genome sequence was obtained for strain LMG 28357T (Gold ID Gp0108260); its DNA G+C content was 30.4%, which is within the range reported for members of the family Flavobacteriaceae (27 to 56 mol%) and which is similar to that of the type strain of A. adventoris (29.0 mol%). Whole-cell fatty acid methyl ester analysis of strain LMG 28357T revealed many branched-chain fatty acids, a typical characteristic of bacteria of the family Flavobacteriaceae and a profile that was similar to that reported for A. adventoris wkB301T. MK6 was the major respiratory quinone, again conforming to bacteria of the family Flavobacteriaceae. The isolates LMG 28357T and LMG 28623 could be distinguished from A. adventoris strains through their oxidase activity. On the basis of phylogenetic, genotypic and phenotypic data, we propose to classify both isolates as representatives of a novel species of the genus Apibacter, Apibacter mensalis sp. nov., with LMG 28357T (=DSM 100903T=R-53146T) as the type strain.

Potential of Bacillus amyloliquefaciens for Biocontrol of Bacterial Wilt of Tomato Incited by Ralstonia solanacearum

Fifty seven rhizobacteria were isolated from rhizospheric soil of wilted tomato plants and among them two strains of rhizobacteria, having better antagonistic and plant growth promoting ability were characterized them as Bacillus amyloliquefaciens DSBA-11 and DSBA-12 based on morphological, biochemical, partial gene sequence analysis of 16S rRNA and fatty acid methyl ester analysis. Antagonistic activity of these strains DSBA-11, DSBA-12 was compared with other Bacillus species such as B. subtilis DTBS-5, B. cereus JHTBS-7, B. pumilus MTCC-7092 strains, against Ralstonia solanacearum race 1, bv 3, phylotype I, inciting bacterial wilt of tomato underin vitro conditions. B. amyloliquefaciens DSBA-11 showed maximum growth inhibition of R. solanacearum (4.91cm2) followed by strains DSBA-12 (3.31cm2) and B. subtilis (3.07 cm2). Moreover, strains DSBA-11 was also have better phosphorus solubilizing ability (42.6 μg/ml) and indole acetic acid (95.4 μg/ml) production than other strains of Bacillus spp. in vitro conditions. Biocontrol efficacy and plant growth ability of these bacterial antagonists was tested against bacterial wilt of tomato cv. Pusa Ruby under glasshouse conditions. Minimum bacterial wilt disease incidenceincultivar Pusa Ruby (17.95%) was recorded in B. amyloliquefaciens DSBA-11followed by B. amyloliquefaciens DSBA-12 after 30 days of inoculation.The bio-control efficacy was higher in B. amyloliquefaciens DSBA -12 treated plants, followed by B. pumilus MTCC- 7092.

Plasma phospholipid pentadecanoic acid, EPA, and DHA, and the frequency of dairy and fish product intake in young children

BackgroundThere is a lack of studies comparing dietary assessment methods with the biomarkers of fatty acids in children.ObjectiveThe objective was to evaluate the suitability of a food frequency questionnaire (FFQ) to rank young children according to their intake of dairy and fish products by comparing food frequency estimates to the plasma phospholipid fatty acids pentadecanoic acid, eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA).DesignCross-sectional data for the present study were derived from the prospective cohort ‘Environmental Triggers of Type 1 Diabetes Study’. Infants were recruited from the Norwegian general population during 2001–2007. One hundred and ten (age 3–10 years) children had sufficient volumes of plasma and FFQ filled in within 2 months from blood sampling and were included in this evaluation study. The quantitative determination of plasma phospholipid fatty acids was done by fatty acid methyl ester analysis. The association between the frequency of dairy and fish product intake and the plasma phospholipid fatty acids was assessed by a Spearman correlation analysis and by investigating whether participants were classified into the same quartiles of distribution.ResultsSignificant correlations were found between pentadecanoic acid and the intake frequency of total dairy products (r=0.29), total fat dairy products (r=0.39), and cheese products (r=0.36). EPA and DHA were significantly correlated with the intake frequency of oily fish (r=0.26 and 0.37, respectively) and cod liver/fish oil supplements (r=0.47 for EPA and r=0.50 DHA). To a large extent, the FFQ was able to classify individuals into the same quartile as the relevant fatty acid biomarker.ConclusionsThe present study suggests that, when using the plasma phospholipid fatty acids pentadecanoic acid, EPA, and DHA as biomarkers, the FFQ used in young children showed a moderate capability to rank the intake frequency of dairy products with a high-fat content and cod liver/fish oil supplements.