Branched-chain Fatty Acids Content Research Articles

Effect of Supplementing a Bacillus Multi-Strain Probiotic to a Post-Weaning Diet on Nutrient Utilisation and Nitrogen Retention of Piglets

Probiotics are suggested to improve pig health, nutrient utilisation, performance, and they may reduce nitrogen (N) pollution. However, the effectivity of a single strain might be different from that of a multi-strain. The study was conducted to investigate the effect of a novel Bacillus multi-strain on nutrient digestibility, energy utilisation, and N retention in weaned piglets using an European diet. The experiment consisted of a control diet (CD) and a supplemented diet (SD). The probiotic used for SD consisted of B. amyloliquefaciens—516 and B. subtilis—541. A total of eight boars/treatment were weaned (day 0; 8.5 kg body weight). Only boars were used to ease the collection of urine. Until day 10, piglets were fed ad libitum and were housed in pairs; from day 11, piglets were fed semi ad libitum (feeding level 3.2× metabolic body weight) and were housed individually. From day 14, faecal and urine were collected twice daily. Piglets were humanely euthanised at day 19 (15.0 kg bodyweight) after which the jejunum, ileum, and colon content were collected. In faeces, the apparent total tract digestibility (ATTD) of, amongst others, DM, organic matter (OM), crude protein (CP), non-starch polysaccharides (NSP), and subsequently net energy (NE) were calculated using titanium dioxide as an indigestible marker. In the jejunum and ileum, the apparent digestibility of CP was estimated, and in the ileum, the apparent AA digestibility was measured. In urine, the N content was measured to determine N retention. The volatile fatty acid (VFA), branched-chain fatty acid (BCFA), and lactic acid content were analysed in the colon and faeces. The apparent CP digestibility in the jejunum and ileum was not affected by treatment (p > 0.05), and no effect was observed on the apparent ileal digestibility of AA (p > 0.05). Supplementation with the multi-strain probiotic improved the ATTD of DM (p = 0.01; +1.3%) and OM (p = 0.02; +1.2%) and tended to improve the ATTD of CP (p = 0.10; +2.2%) and NSP (p = 0.07; +1.9%). The multi-strain probiotic also improved the NE value (p = 0.02; +0.2 MJ/kg DM) and improved N retention (p = 0.05; +1.6%). Supplementation did not influence the VFA, BCFA, and lactic acid content in the faeces (p > 0.05). However, in the colon, supplementation did influence the lactic acid content (lower; p = 0.01) and tended to influence the valeric acid content (higher; p = 0.09). In conclusion, results from the current study suggest that the multi-strain probiotic has the potential to contribute to improve nutrient efficiency in weaned piglets. More research needs to be conducted to identify the impact of the improved nutrient utilisation on gut health in post-weaned pigs as well as environmental pollution.

The effects of protease, xylanase, and xylo-oligosaccharides on growth performance, nutrient utilization, short-chain fatty acids, and microbiota in Eimeria-challenged broiler chickens fed low-protein diet

A total of 392 Cobb 500 off-sex male broiler chicks were used in a 21-day experiment to study the effect of protease, xylanase, and xylo-oligosaccharides (XOS) on improving growth performance, nutrient utilization (ileal digestibility and total tract retention), gene expression of nutrient transporters, cecal short-chain fatty acids (SCFAs), and microbiota profile of broilers challenged with Eimeria spp. Chicks at 0-day old were allocated to 8 treatments in a 4×2 factorial arrangement: 1) corn-soybean meal diet with no enzyme (Con); 2) Con plus 0.2 g/kg protease alone (PRO); 3) Con plus 0.2 g/kg protease combined with 0.1 g/kg xylanase (PRO+XYL); or 4) Con plus 0.5 g/kg xylo-oligosaccharides (XOS); with or without Eimeria challenge. The 4 diets were formulated to be marginally low in crude protein (183 g/kg). Challenged groups were inoculated with a solution containing E. maxima, E. acervulina, and E. tenella oocysts on d 15. Eimeria depressed (P < 0.01) growth performance and nutrient utilization. Supplemental protease improved (P < 0.05) body weight gain and feed intake in the prechallenge phase (d 0-15) but had no effect during the infection period (d 15-21). There was no interaction between infection and feed supplementation for nutrient utilization. The supplementations of either PRO or XOS alone increased (P < 0.01) total tract retention of Ca and tended (P < 0.1) to improve total tract retention of N, P, AME, and AMEn. Eimeria decreased (P < 0.05) expressions of GLUT2, GLUT5, PepT1, ATP2B1, CaSR, Calbidin D28K, NPT2, and ZnT1 but increased (P < 0.01) expression of GLUT1. XOS supplementation increased (P < 0.05) ATP2B1 expression. Protease decreased (P < 0.05) isobutyrate concentration in unchallenged treatments but not in challenged treatments. Eimeria decreased (P < 0.01) cecal saccharolytic SCFAs acetate and propionate but increased (P < 0.01) branched-chain fatty acid isovalerate. The supplementation of PRO+XYL or XOS increased (P < 0.05) cecal butyrate or decreased cecal isobutyrate concentrations, respectively. PRO+XYL and XOS decreased cecal protein levels in unchallenged birds but not challenged ones. Eimeria challenge significantly (P < 0.05) decreased the microbial richness (Observed features) and diversity (Shannon index and phylogenetic diversity) and changed the microbial composition by reducing the abundance of certain bacteria, such as Ruminococcus torques, and increasing the abundance of others, such as Anaerostipes. In contrast, none of the additives had any significant effect on the cecal microbial composition. In conclusion, PRO or XOS supplementation individually improved nutrient utilization. All the additives decreased the cecal content of branched-chain fatty acids, consistent with decreased cecal N concentration, although the effects were more pronounced in unchallenged birds. In addition, none of the feed additives impacted the Eimeria-induced microbial perturbation.

Identification and quantification of branched-chain fatty acids and odd-chain fatty acids of mammalian milk, dairy products, and vegetable oils using GC/TOF-MS

Branched-chain fatty acids (BCFAs) and odd-chain fatty acids (OCFAs) in mammalian milk, dairy products, and vegetable oils were investigated; 72 fatty acids, including 16 BCFAs and 10 OCFAs, were identified and quantified using GC/TOF-MS. Human milk showed the lowest content of BCFA (0.4% of total FA) and the content in other milk ranged from 2 to 6%, of which ai17:0, i17:0, ai15:0 and i15:0 were most abundant. Differences in the content of OCFA in mammalian milk trended similarly to differences in BCFA content. The molecular species of BCFA in cow milk-derived dairy products were identical, but the content was different, with higher content in fermented dairy products. Apparent differences in BCFA and OCFA composition were found in infant formulas, strongly related to their fat sources. Vegetable oils contained no BCFA and a small amount of OCFA. A scientific basis for further investigation of high-BCFA and OCFA dairy products is provided.

Fatty Acid and Antioxidant Profile of Eggs from Pasture-Raised Hens Fed a Corn- and Soy-Free Diet and Supplemented with Grass-Fed Beef Suet and Liver.

There is increasing interest in using grass-fed beef (GFB) by-products to augment the nutrient profile of eggs among local pasture-raising systems in the US. The objective of this study was to characterize egg yolk fatty acid and antioxidant profiles using eggs from pasture-raised hens fed a corn- and soy-free diet and supplemented with GFB suet and liver compared to eggs from pasture-raised hens fed a corn and soy layer hen feed and commercially obtained cage-free eggs. The egg yolk vitamin and mineral profile was also assessed by a commercial laboratory. Both pasture-raised groups had twice as much carotenoid content, three times as much omega-3 fatty acid content, and a 5-10 times lower omega-6:omega-3 fatty acid ratio compared to the cage-free eggs (p &lt; 0.001). Eggs from hens fed a corn- and soy-free feed and GFB by-products had half as much omega-6 fatty acid content and five times more conjugated linoleic acid, three times more odd-chain fatty acid, and 6-70 times more branched-chain fatty acid content (p &lt; 0.001). Feeding pasture-raised hens GFB suet and liver reduces agricultural waste while producing improved egg products for consumers, but further research is needed to quantify optimal supplementation levels and the efficacy of corn- and soy-free diets.

Effect of rice bran and microorganism as additives in pearl millet silage

ABSTRACT This study aimed to evaluate rice bran (RB) and microbial inoculation effects on pearl millet silage fermentative losses, fermentation profile, nutrients recovery, chemical composition, in vitro degradation and aerobic stability. Seventy-two experimental silos (PVC buckets with 28 cm diameter and 25 cm height) were used to evaluate nine treatments, from a 3 × 3 factorial arrangement: (I) levels of RB: 0, 100 and 200 g/kg as-fed; and (II) microbial inoculation: (1) Control (CON): no inoculation; (2) LPPP: inclusion 1.04 × 105 colony forming units (CFU) of Lactobacillus plantarum and 1.04 × 105 CFU of Pediococcus pentosaceus per g; and (3) LB: inclusion 1.6 × 105 CFU of Lactobacillus buchneri per g. The silage microbial inoculant inclusion had no effect on silage, regardless of RB addition. Rice bran increased silage pH and decreased ammonia-N, propionic, butyric and branched-chain fatty acids content. Rice bran decreased gas losses and recovery of DM, OM, CP and ADF, whereas increased digestible DM recovery. The addition of RB reduced OM, NDF, ADF and ADL silage content, and increased silage DM, CP and EE, and NDF degradation. Therefore, RB increases digestible DM recovery, and improves silage fermentation and nutritional value.

Identification of High and Low Branched-Chain Fatty Acid–Producing Phenotypes in Holstein Cows following High-Forage and Low-Forage Diets in a Crossover Designed Trial

Branched-chain fatty acids (BCFAs) are rumen-derived fatty acids comprising ∼2% of bovine-milk fatty acids. BCFAs possess anti-inflammatory properties and enriching the BCFA content of bovine milk may provide human health benefits. We determined whether forage content impacts the BCFA content of milk from Holstein cows and identified fatty acid phenotypes in high vs. low BCFA-containing milks. Holstein cows (n = 62), fed for 67 d in a crossover design, consumed a diet with high forage and low concentrate (HF:C) and a diet with low forage and high concentrate (LF:C). Milk samples were collected at the end of each treatment period and fatty acid content determined. Paired t-tests, 1-factor ANOVA, sparse partial least-squares discriminant analysis (sPLSDA), and Pearson's correlation analysis were used to analyze the data. The total milk fatty acid concentration for cows fed the HF:C diet was greater than that of cows fed the LF:C diet (4.2 ± 0.7 g/100 mL vs. 3.9 ± 0.9 g/100 mL). sPLSDA demonstrated separation of the dietary treatments, with BCFAs and odd-chain fatty acids as primary determinants. Total BCFA content in milk fat was elevated by HF:C intake compared with LF:C intake (1.80 vs. 1.68%). Quintile separation of high vs. low BCFA milks resulted in 4 groups: HF:C /low BCFAs, HF:C /high BCFAs; LF:C /low BCFAs, and LF:C /high BCFAs. Milks from the high BCFA quintiles had lower palmitic acid content (29.6% vs. 34.4%) but higher oleic acid content than milks from the low BCFA quintiles (19.7% vs. 17.0%). Some cows were identified as high BCFA producers or low BCFA producers regardless of diet. BCFA content of milk is diet-sensitive but variation in responses exists. The potential to produce milk with high BCFA content and lower SFA content needs further study.

Branched-chain fatty acids in the vernix caseosa and meconium of infants born at different gestational ages.

The functional lipid components found in breast milk, vernix caseosa, and meconium are Branched‐chain Fatty Acids (BCFA). The goal of this study was to establish the existence of BCFA in vernix and meconium in infants born at different gestational ages. TLC plates and GC‐MS were examined for the lipids in vernix caseosa and meconium. The results indicated that there were nine BCFA in vernix caseosa, including iso‐12:0, anteiso‐13:0, iso‐14:0, iso‐15:0, anteiso‐15:0, iso‐16:0, anteiso‐17:0, iso‐18:0, and iso‐20:0. Five BCFA (iso‐12:0, anteiso‐13:0, iso‐14:0, iso‐15:0, and anteiso‐15:0) were not contained in the meconium, suggesting that some of the BCFA may be digested and consumed by infants. In the vernix caseosa, the content of BCFA in triacylglycerol (TAG) and free fatty acid (FFA) was 15.59% and 11.82%, respectively. The vernix caseosa's wax ester fraction contained the highest content of BCFA, reaching up to 16.81%. The carbon chain length of fatty acids (FA) ranged from 12 to 24 in the vernix caseosa and 14 to 22 in meconium samples. The gestational age was likely to affect BCFA concentrations, with the vernix caseosa and meconium BCFA content being significantly higher in full‐term infants than in preterm infants (p < .001). Further research is required into the relationship between BCFA and gut microbiotas.

Branched-Chain Fatty Acids-An Underexplored Class of Dairy-Derived Fatty Acids.

Dairy fat and its fatty acids (FAs) have been shown to possess pro-health properties that can support health maintenance and disease prevention. In particular, branched-chain FAs (BCFAs), comprising approximately 2% of dairy fat, have recently been proposed as bioactive molecules contributing to the positive health effects associated with the consumption of full-fat dairy products. This narrative review evaluates human trials assessing the relationship between BCFAs and metabolic risk factors, while potential underlying biological mechanisms of BCFAs are explored through discussion of studies in animals and cell lines. In addition, this review details the biosynthetic pathway of BCFAs as well as the content and composition of BCFAs in common retail dairy products. Research performed with in vitro models demonstrates the potent, structure-specific properties of BCFAs to protect against inflammation, cancers, and metabolic disorders. Yet, human trials assessing the effect of BCFAs on disease risk are surprisingly scarce, and to our knowledge, no research has investigated the specific role of dietary BCFAs. Thus, our review highlights the critical need for scientific inquiry regarding dairy-derived BCFAs, and the influence of this overlooked FA class on human health.

Branched-Chain Fatty Acid Content Modulates Structure, Fluidity, and Phase in Model Microbial Cell Membranes.

Recent progress in understanding the importance and origins of lipid rafts in microbial cell membranes has focused attention on membranes containing branched-chain fatty acids. The working hypothesis is that branched fatty acids increase the fluidity of the bilayer, analogous to unsaturated fatty acids in membranes of higher organisms. Here, we perform a series of 7 μs long atomistic simulations on biomimetic, branched-chain lipid containing bilayer patches, systematically varying the amount of the straight-chain fatty acid component, n16:0, from 7.0 to 47.3 mol %. The simulations reveal thickening and ordering of the bilayer as well as higher bilayer viscosity and bending modulus with increasing n16:0 content, thus providing quantitative support that branched fatty acids increase the bilayer fluidity. A sharp transition in these properties is observed at ∼20% n16:0 content, resembling a phase change. The simulations provide the first access to ordered and disordered phases in a bacterial cell membrane mimic containing branched-chain lipids. Granted several assumptions, a comparison of these phases provides estimates of physical properties such as hydrophobic mismatch (∼1.2 Å), difference in bending moduli (∼15.7 kBT), and the line tension (∼0.6 pN) for a putative lipid raft in the cell membrane of an organism such as Bacillus subtilis or Staphylococcus aureus.

Exercise and Conjugated Linoleic Acid Supplementation Induce Changes in the Composition of Liver Fatty Acids.

Exercise and supplementation with conjugated linoleic acid (CLA) are used to reduce body weight and to improve health. Applied together, they may exert a synergistic effect. However, the effects of exercise and CLA supplementation on liver metabolism are poorly understood. The aim of this study was to examine the influence of exercise and CLA supplementation on fatty acids (FA) composition in mouse liver. We analyzed 44 of known FAs of this organ by gas chromatography-mass spectrometry. Our results demonstrated that exercise contributed to a decrease in odd-chain FA and an increase in n-6 polyunsaturated FA contents. In turn, CLA stimulated an increase in branched-chain FAs and n-3 polyunsaturated FAs. Exercise combined with CLA supplementation caused a substantial increase in branched-chain FA content and a slight increase in n-6 polyunsaturated FAs. The elevated liver content of branched-chain FAs after the exercise combined with CLA supplementation, as well as the increase in n-3 polyunsaturated FAs after CLA may be favorable since both these FA groups were previously shown to produce health benefits. However, the expression pattern of enzymes involved in fatty acid synthesis did not match the changes in FA composition. Thus, the mechanism of exercise- and CLA-induced changes in liver FA profile is yet to be established. Also, the consequences of CLA- and/or exercise-induced changes in the composition of liver FAs need to be elucidated.

High levels of branched chain fatty acids in nātto and other Asian fermented foods

The mean intake (500 mg/day) of branched chain fatty acids (BCFA) in western countries is from dairy products and beef. We hypothesized that Asian countries with low dairy consumption have an alternative source of BCFA and report the characterization of nātto and selected fermented foods for BCFA content. Nātto BCFA averaged 0.6 (range 0.21–1.43) mg BCFA per g natto (wet weight) and was highly variable. Nātto major BCFA are C14-17 iso- and anteiso-BCFA similar to fluid milk. BCFA concentrations were 1.00 ± 0.64%, 1.63 ± 0.72% and 0.65 ± 0.07%, of total fatty acids in nātto, shrimp paste and fish sauce, respectively. In contrast, saturates, monounsaturates, and major polyunsaturates were more constant (coefficient of variation = 21%, 26% and 4% compared to 64% for nātto BCFA). Detection of fatty acid ethyl esters were confirmed in miso and found in homemade kimchi. Habitual nātto and/or fermented seafood consumption could support BCFA intakes similar to dairy consumption.

Heptadecanoic acid as an indicator of BCFA content in sheep fat

Branched chain fatty acids (BCFAs) are associated with ‘mutton flavour’ found in cooked sheep meat aroma. Three BCFAs, 4-methyloctanoic (MOA), 4-ethyloctanoic (EOA) and 4-methylnonanoic (MNA) acids, have been implicated as the main compounds responsible for ‘mutton flavour’. Animal age can also contribute to increasing BCFA concentration. Heptadecanoic acid (C17:0 FA) also increases with animal age in sheep. Using published data, a linear association has been found to exist between C17:0 FA with MOA and MNA in sheep fat, with C17:0 FA increasing proportionally with these two compounds. No association was found between C17:0 FA and EOA. As C17:0 FA is present in sheep fat in relatively higher proportions compared to BCFAs, it has potential to be used as a proxy for MOA and MOA as well as ‘mutton flavour’.

Impact of Branched-Chain Amino Acid Catabolism on Fatty Acid and Alkene Biosynthesis in Micrococcus luteus.

Micrococcus luteus naturally produces alkenes, unsaturated aliphatic hydrocarbons, and represents a promising host to produce hydrocarbons as constituents of biofuels and lubricants. In this work, we identify the genes for key enzymes of the branched-chain amino acid catabolism in M. luteus, whose first metabolic steps lead also to the formation of primer molecules for branched-chain fatty acid and olefin biosynthesis, and demonstrate how these genes can be used to manipulate the production of specific olefins in this organism. We constructed mutants of several gene candidates involved in the branched-chain amino acid metabolism or its regulation and investigated the resulting changes in the cellular fatty acid and olefin profiles by GC/MS. The gene cluster encoding the components of the branched-chain α-keto acid dehydrogenase (BCKD) complex was identified by deletion and promoter exchange mutagenesis. Overexpression of the BCKD gene cluster resulted in about threefold increased olefin production whereas deletion of the cluster led to a drastic reduction in branched-chain fatty acid content and a complete loss of olefin production. The specificities of the acyl-CoA dehydrogenases of the branched amino acid degradation pathways were deduced from the fatty acid and olefin profiles of the respective deletion mutant strains. In addition, growth experiments with branched amino acids as the only nitrogen source were carried out with the mutants in order to confirm our annotations. Both the deletion mutant of the BCKD complex, responsible for the further degradation of all three branched-chain amino acids, as well as the deletion mutant of the proposed isovaleryl-CoA dehydrogenase (specific for leucine degradation) were not able to grow on leucine in contrast to the parental strain. In conclusion, our experiments allow the unambigous assignment of specific functions to the genes for key enzymes of the branched-chain amino acid metabolism of M. luteus. We also show how this knowledge can be used to engineer the isomeric composition and the chain lengths of the olefins produced by this organism.

The impact of lactation and gestational age on the composition of branched-chain fatty acids in human breast milk.

Breast milk consumption reduces the incidence of necrotizing enterocolitis (NEC) in preterm infants compared to formula. Branched-chain fatty acids (BCFAs) are present in breast milk but not in most formulas intended for preterm infants. We aimed to determine the composition of BCFAs in the breast milk of mothers with preterm infants, and to understand the impact of gestational age at birth and stage of lactation on BCFA content. The main BCFAs in preterm breast milk were iso-14:0, iso-15:0, anteiso-15:0, iso-16:0, iso-17:0, and anteiso-17:0. Breast milk BCFAs as a percent of total fatty acids (g per 100 g, %) were significantly different across lactation stages, with the highest concentration in colostrum, followed by transitional and mature breast milk (median: 0.41, 0.31, and 0.28%, respectively, p < 0.05). Lower BCFAs in preterm breast milk compared to term breast milk may have been related to maternal intake, or the ability of the mammary gland to extract BCFA from plasma, or differences in mammary gland BCFA synthesis. BCFAs were mainly in the sn-2 position (52-65%), similar to palmitic acid. Overall, preterm and term breast milk BCFAs were similar and showed specific concentration patterns, resembling 16:0 with respect to sn-2 positional distribution. BCFAs were reduced with lactation stage, similar to highly unsaturated fatty acids.

Branched chain fatty acids positional distribution in human milk fat and common human food fats and uptake in human intestinal cells

Branched chain fatty acids (BCFA) are components of common food fats but little is known about their positional distribution on triacylglycerols (TAG). Human milk fat, mature milk of cows and goats, and several fish oil products available in China were evaluated for BCFA content in the sn-2 position. Compared to the sn-1/sn-3 (α) positions, BCFA are enriched in sn-2 (β) position of human milk (68% of BCFA), but randomly distributed among the sn-2 and sn-1/sn-3 positions of cow and goat milk. Fish oil TAG sn-2 BCFA were consistently depleted compared to the sn-1/sn-3 positions (5.2–11.9% of BCFA). Human intestinal epithelial cells were incubated with human milk sn-2 MAG, or fatty acids liberated from human milk sn-2 MAG. Cells incorporated more BCFA as sn-2 MAG than as FFA. The stereochemical positioning of BCFA in human milk fat may be unique compared to other BCFA sources, and mediate selective absorption of BCFA.

Saturated Branched Chain, Normal Odd-Carbon-Numbered, and n-3 (Omega-3) Polyunsaturated Fatty Acids in Freshwater Fish in the Northeastern United States.

The fatty acid profiles of wild freshwater fish are poorly characterized as a human food source for several classes of fatty acids, particularly for branched chain fatty acids (BCFA), a major bioactive dietary component known to enter the US food supply primarily via dairy and beef fat. We evaluated the fatty acid content of 27 freshwater fish species captured in the northeastern US with emphasis on the BCFA and bioactive polyunsaturated fatty acids (PUFA) most associated with fish, specifically n-3 (omega-3) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Mean BCFA content across all species was 1.0 ± 0.5% (mean ± SD) of total fatty acids in edible muscle, with rainbow smelt (Osmerus mordax) and pumpkinseed (Lepomis gibbosus) the highest at >2% BCFA. In comparison, EPA + DHA constituted 28% ± 7% of total fatty acids. Across all fish species, the major BCFA were iso-15:0, anteiso-15:0, iso-16:0, iso-17:0 and anteiso-17:0. Fish skin had significantly higher BCFA content than muscle tissues, at 1.8% ± 0.7%, but lower EPA and DHA. Total BCFA in fish skins was positively related with that in muscle (r2 = 0.6). The straight chain saturates n-15:0 and n-17:0 which have been identified previously as markers for dairy consumption were relatively high with means of 0.4% and 0.6%, respectively, and may be an underappreciated marker for seafood intake. Consuming a standardized portion, 70 g (2.5 oz), of wild freshwater fish contributes only small amounts of BCFA, 2.5-24.2 mg, to the American diet, while it adds surprisingly high amounts of EPA + DHA (107 mg to 558 mg).

Effect of Storage Period on the Changes of Odorous Compound Concentrations and Bacterial Ecology for Identifying the Cause of Odor Production from Pig Slurry.

Odor from buildings where pigs are housed is generated by anaerobic fermentation of undigested materials in pig slurry stored for several weeks in pit. The objective of this study was to investigate the effect of storage period on the level of odorous compounds in pig slurry and on its bacterial community. A slurry sample (15 L) was taken from the pit of a finisher pig building and incubated in acryl chambers for six- weeks. Slurry for analysis was sampled every two-week. Levels of odorous compounds in the slurry sample were drastically changed after two weeks of storage period; levels of phenols and short chain fatty acids (SCFAs) were decreased (P<0.05), whereas indoles and branched-chain fatty acids (BCFAs) were increased (P<0.05). Among dominant bacteria, Bacteroides and Porphyromonadacese_uc_g revealed a strong positive correlation with the levels of phenols and SCFAs. Populations of AC160630_g, Acholeplasmatales_uc_g, Mollicutes_uc_g and Cloacamonas_f_uc_g positively correlated with indole and BCFAs content. Taken together, levels of odorous compounds were increased after two weeks of storage, possibly because of changes in the predominant bacterial groups to those that use protein as a carbon source in the hypo-carbohydrate conditions.

Broad substrate specificity of phosphotransbutyrylase from Listeria monocytogenes: A potential participant in an alternative pathway for provision of acyl CoA precursors for fatty acid biosynthesis

Listeria monocytogenes, the causative organism of the serious food-borne disease listeriosis, has a membrane abundant in branched-chain fatty acids (BCFAs). BCFAs are normally biosynthesized from branched-chain amino acids via the activity of branched chain α-keto acid dehydrogenase (Bkd), and disruption of this pathway results in reduced BCFA content in the membrane. Short branched-chain carboxylic acids (BCCAs) added as media supplements result in incorporation of BCFAs arising from the supplemented BCCAs in the membrane of L. monocytogenes bkd mutant MOR401. High concentrations of the supplements also effect similar changes in the membrane of the wild type organism with intact bkd. Such carboxylic acids clearly act as fatty acid precursors, and there must be an alternative pathway resulting in the formation of their CoA thioester derivatives. Candidates for this are the enzymes phosphotransbutyrylase (Ptb) and butyrate kinase (Buk), the products of the first two genes of the bkd operon. Ptb from L. monocytogenes exhibited broad substrate specificity, a strong preference for branched-chain substrates, a lack of activity with acetyl CoA and hexanoyl CoA, and strict chain length preference (C3–C5). Ptb catalysis involved ternary complex formation. Additionally, Ptb could utilize unnatural branched-chain substrates such as 2-ethylbutyryl CoA, albeit with lower efficiency, consistent with a potential involvement of this enzyme in the conversion of the carboxylic acid additives into CoA primers for BCFA biosynthesis.

Branched chain fatty acid content of edible portions of northeastern U.S. freshwater fish

IntroductionBranched chain fatty acids (BCFA) are major components of skin/membranes across various mammals and bacterial genera. BCFA are bioactive and alter gut microbiota. Intake of BCFA, mainly from dairy/beef was estimated at 500 mg per day in US. Our goal was to investigate BCFA levels in common freshwater fish species.MethodsWe captured over 20 fish species from several lakes and streams in New York and Pennsylvania. We dissected larger fish and sampled muscle tissue from dorsal and caudal fins as well as the belly and the skin. Smaller fish were entirely grinded before processing. All samples were extracted, saponified and methylated. BCFA profiles were analyzed using gas chromatography mass spectrometry.ResultsBCFA content varied across species and ranged from 0.6–2.7%, w/w of total fatty acids in fish muscle samples, with an overall mean of 1.2%. Some low trophic level fishes, such as killifish, rainbow smelt, and juvenile pumpkinseed, had over 2% BCFA while some common game fish species white perch, yellow perch, smallmouth bass, and walleye had about 1% BCFA, w/w or less of total fatty acids. A few low trophic level fishes such as fantail darter in the same perch family as yellow perch and walleye had low BCFA levels, suggesting influences related to phylogenetic structure. Skin contained between 1.2–3.2% BCFA, w/w of total fatty acids. Across all species sampled, skin samples had higher BCFA levels compared to muscle samples. The overall fatty acid profile, including BCFA was similar across muscles from different parts of a fish. The major BCFA in fish are iso‐15:0, anteiso‐15:0, iso‐16:0, iso‐17:0 and anteiso‐17:0. Trace amounts of iso‐13:0, anteiso‐13:0, iso‐14:0, iso‐18:0, anteiso‐19:0 and iso‐20:0 were also detected.ConclusionMany low‐trophic‐level fishes had BCFA levels comparable with dairy products and their skin samples showed higher BCFA content than muscle.Support or Funding InformationNIH R01 AT007003

Content and Composition of Branched-Chain Fatty Acids in Bovine Milk Are Affected by Lactation Stage and Breed of Dairy Cow.

Dairy products contain bioactive fatty acids (FA) and are a unique dietary source of an emerging class of bioactive FA, branched-chain fatty acids (BCFA). The objective of this study was to compare the content and profile of bioactive FA in milk, with emphasis on BCFA, among Holstein (HO), Jersey (JE), and first generation HO x JE crossbreeds (CB) across a lactation to better understand the impact of these factors on FA of interest to human health. Twenty-two primiparous cows (n = 7 HO, n = 7 CB, n = 8 JE) were followed across a lactation. All cows were fed a consistent total mixed ration (TMR) at a 70:30 forage to concentrate ratio. Time points were defined as 5 days in milk (DIM), 95 DIM, 185 DIM, and 275 DIM. HO and CB had a higher content of n-3 FA at 5 DIM than JE and a lower n-6:n-3 ratio. Time point had an effect on the n-6:n-3 ratio, with the lowest value observed at 5 DIM and the highest at 185 DIM. The content of vaccenic acid was highest at 5 DIM, yet rumenic acid was unaffected by time point or breed. Total odd and BCFA (OBCFA) were higher in JE than HO and CB at 185 and 275 DIM. Breed affected the content of individual BCFA. The content of iso-14:0 and iso-16:0 in milk was higher in JE than HO and CB from 95 to 275 DIM. Total OBCFA were affected by time point, with the highest content in milk at 275 DIM. In conclusion, HO and CB exhibited a higher content of several bioactive FA in milk than JE. Across a lactation the greatest content of bioactive FA in milk occurred at 5 DIM and OBCFA were highest at 275 DIM.