Branched-chain Fatty Acids Research Articles

A Novel Prebiotic Fibre Blend Supports the Gastrointestinal Health of Senior Dogs.

Senior pets can suffer from a wide range of age-related diseases that can impact the quality of life for the pet and the relationship between a pet and their owner. Dietary fibre plays a key role in shaping the gastrointestinal health in mammalian species. The aim of this study was to investigate the impact of a novel prebiotic fibre blend containing sugar beet pulp, galacto-oligosaccharides, and cellulose on the health of senior dogs when fed on top of a background commercial dry diet. Thirty-two dogs aged >8 years received the prebiotic fibre blend as a dietary topper for 21 days on top of a nutritionally complete diet using a cross-over study design. The prebiotic fibre blend improved the gastrointestinal health of senior dogs as measured through improved faecal quality scores, a reduction in faecal pH, changes to the taxonomic composition of the gut, and a reduction in faecal branched-chain fatty acids, which are markers for proteolytic degradation. Broader systemic measures, such as changes to serum cytokines, were not impacted by the prebiotic fibre blend. In conclusion, a novel prebiotic fibre blend containing sugar beet pulp, galacto-oligosaccharides, and cellulose improved the gastrointestinal health of senior dogs and could have a range of potential future dietary applications.

Comprehensive evaluation of the prebiotic properties of Dendrobium officinale polysaccharides, β-glucan, and inulin during in vitro fermentation via multi-omics analysis

Dietary fiber is crucial for human health mainly due to its impact on gut microbiota structure and metabolites. This study aimed to investigate the impact of Dendrobium officinale polysaccharides (DOP) and two common fibers (β-glucan and inulin) on the gut microbiome structure and metabolic profile in vitro. Fecal samples were obtained from 30 healthy volunteers, which were then individually subjected to fermentation with each type of fiber. The results revealed that all fibers were efficiently degraded by gut microbiota, with DOP exhibiting a slower fermentation rate compared to β-glucan and inulin. The fermentation of all fibers led to a significant increase in the production of short-chain fatty acids (SCFAs) and a reduction in branched-chain fatty acids (BCFAs), sulfides, phenols, and indole. Moreover, the abundance of unclassified Enterobacteriaceae, which was positively correlated with sulfide, phenols, and indole levels, was significantly reduced by all fibers. Additionally, DOP specifically promoted the growth of Parabacteroides, while β-glucan and inulin promoted the growth of Bifidobacterium and Faecalibacterium. Taken together, these findings enhance our understanding of the role of DOP, β-glucan, and inulin in modulating gut microbiota and metabolites, where the fermentation with fecal bacteria from different volunteers could provide valuable insights for personalized therapeutic approaches.

Effect of providing citrus pulp-integrated diet on fecal microbiota and serum and fecal metabolome shifts in crossbred pigs

The study aimed to assess the impact of dehydrated citrus pulp (DCP) on growth performance, fecal characteristics, fecal bacterial composition (based on 16S rRNA analysis), and fecal and serum metabolomic profiles in crossbred pigs. 80 finishing pigs Duroc × (Landrace × Large White) were fed either a control diet (C) or a diet with 240 g/kg DCP (T) for six weeks. Including DCP in diets tended to decrease feed intake, increased (p < 0.05) the concentrations of acetic and heptanoic acids and decreased (p < 0.05) fecal butyric and branched-chain fatty acid concentrations in feces. Animals fed DCP exhibited a lower abundance of the genera Clostridium and Romboutsia, while Lachnospira significantly increased. Orthogonal partial least squares discriminant analysis plotted a clear separation of fecal and serum metabolites between groups. The main discriminant fecal metabolites were associated with bacterial protein fermentation and were downregulated in T-fed pigs. In serum, DCP supplementation upregulated metabolites related to protein and fatty acids metabolism. In conclusion, the addition of DCP as an environmentally friendly source of nutrients in pig diets, resulted in modifications of fecal bacterial composition, fermentation patterns, and overall pig metabolism, suggesting improvements in protein metabolism and gut health.

Enhanced production of anteiso-branched chain fatty acids rich bacterial oil using Lentibacillus salarius strain BPIITR under furan aldehydes induced oxidative stress

Monomethyl branched-chain fatty acids (mBCFAs) are highly valuable compounds in the oleochemical industry, with diverse industrial applications including pharmaceuticals, biofuels and lubricants. In the current study, the medium components of L. salarius BPIITR were statistically optimized to produce mBCFAs-rich lipid, yielding up to 227.16 ± 16.2 mg/g dcw. The supplementation of 2-methyl butyric acid increased the lipid content to 276.42 ± 7.46 mg/g dcw, with anteiso fatty acids up to 87.59 ± 1.56 % w/w of the total lipid. The addition of 5-hydroxymethyl furfural (5-HmF) and furfural induced oxidative stress, resulting in the synthesis of lipids rich in anteiso-mBCFAs. Furfural supplementation led to a lipid accumulation of 256.49 ± 7.91 mg/g dcw, whereas with 5-HmF, it increased to 284.75 ± 8.98 mg/g dcw. Furthermore, the elongation of mBCFAs from C15:0 to C17:0 was significantly affected during oxidative stress. This study suggests that under oxidative stress conditions, the synthesis of anteiso fatty acids is favored over iso-fatty acids in L. salarius BPIITR. Additionally, value-added products such as trehalose and furan carboxylic acids were identified as potential co-products in this study, with trehalose reaching up to 255.84 ± 8.04, 163.65 ± 20.94, and 217.46 mg/g dcw in the presence of 2-methylbutyric acid, furfural, and 5-HmF, respectively. The concentration of 2-furan carboxylic acid and 2,5-furan dicarboxylic acids in the fermentation broth reached up to 4.37 ± 0.28 g/L and 2.48 ± 0.35 g/L, respectively. The utilization of short-chain fatty acids and furan aldehydes not only provided a cost-effective carbon source but also modulated the metabolic flux towards specific mBCFAs rich bacterial oil production.

Biomarkers for cheese authentication by detailed and fast gas chromatographic profiling of triacylglycerol fatty acids.

Unsaturated fatty acid isomers and odd- and branched-chain fatty acids (OBCFAs) in milk triacylglycerols (TAGs) can be quantitated using gas chromatography (GC), providing access to biomarkers of animal species, breeds, diet, geographic origin, and environmental conditions. Such analysis requires expensive cyanopropyl siloxane or ionic liquid columns of at least 50m in length, which increases the elution time. Aiming to use GC for cheese authentication and characterization while keeping the experiment time short and maintaining a good separation between fatty acid (FA) isomers, we considered using a 30m polyethylene glycol-2-nitroterephthalate column. The FAs thus quantitated allowed the discovery of specific biomarkers for the origins of cheese varieties highly consumed in several countries. In addition, the simple and multivariate correlations we found between FAs in the cheese TAG matrix were alternative means for characterization and authentication purposes.

Impact of parity on the profile of fatty acids with potential importance for human health in camel (Camelus dromedarius) milk

The aim of the present study was to investigate the influence of parity on the proportion of fatty acids with potential health significance in camel milk fat. Milk samples were collected from four multiparous and four primiparous camels in intensive husbandry at mid-lactation stage. Fatty acid methyl esters were analysed by gas chromatography in conjunction with a flame ionisation detector. Parity had a significant effect on fat and total solids content (p&lt;0.05); mutiparous camels produced milk with lower fat and total solids content. In contrast, protein and ash content were not affected by parity (p&gt;0.05). Primipara camels produced milk richer (p&lt;0.05) in odd and branched chain fatty acids, monounsaturated fatty acids, C17:0, C15:0, omega-3 fatty acids, oleic acid, vaccenic acid and α-linolenic acid, while poorer (p&lt;0.05) for total short-chain fatty acids, total medium-chain fatty acids, total saturated fatty acids, C6:0, C10:0, C14:0, c9-C16:0, C16:0 and iso-C16:0. In addition, the desaturation index for C16, which is an indicator of Δ9-desaturase activity, and the atherogenicity index were lower in primiparous camels (p&lt;0.05). Parity had no effect on conjugated linoleic acid content (p&gt;0.05). Overall, the milk fat of primiparous camels appears to have a relatively high nutritional value, as it contained higher levels of beneficial fatty acids and lower levels of saturated fatty acids.

Catabolism of fats and branched-chain amino acids in children with Type 1 diabetes: Association with glycaemic control and total daily insulin dose.

Hyperglycaemia in Type 1 diabetes (T1D) results from an absolute insulin deficiency. However, insulin resistance (IR) may exacerbate glycaemic instability in T1D and contribute to long-term cardiovascular complications. We previously showed that IR in teenagers with obesity is associated with sex-dependent derangements in the catabolism of branched-chain amino acids (BCAA) and fatty acids. Here we hypothesized that byproducts of BCAA and fatty acid metabolism may serve as biomarkers or determinants of glycaemic control and IR in prepubertal or early pubertal children with T1D. Metabolites, hormones and cytokines from fasting blood samples were analysed in 28 children (15 females, 13 males; age 6-11 years) with T1D. Principal components analysis (PCA) and multiple linear regression models were used to correlate metabolites of interest with glycaemic control, total daily insulin dose (TDD, units/kg/d), adiponectin and the triglyceride (TG) to high-density lipoprotein (HDL) ratio. Males and females were comparable in age, BMI-z, insulin sensitivity, glycaemic control, inflammatory markers, BCAAs and C2/C3/C5-acylcarnitines. The majority of components retained in PCA were related to fatty acid oxidation (FAO) and BCAA catabolism. HbA1c correlated positively with Factor 2 (acylcarnitines, incomplete FAO) and Factor 9 (fasting glucose). TDD correlated negatively with C3 and C5 and Factor 10 (BCAA catabolism) and positively with the ratio of C2 to C3 + C5 and Factor 9 (fasting glucose). These findings suggest that glucose intolerance in prepubertal or early pubertal children with T1D is accompanied by incomplete FAO while TDD is associated with preferential catabolism of fats relative to amino acids.

Electrospray alginate microgels co-encapsulating degraded Konjac glucomannan and quercetin modulate human gut microbiota in vitro

Alginate microgels co-encapsulating degraded Konjac glucomannan (KGM60) underwent in vitro fecal fermentation and their effects on human microbiota and metabolites were investigated. KGM60 delayed quercetin release and enhanced phenolic metabolites production. Microgels co-encapsulating KGM60 and quercetin increased linear short chain fatty acid but decreased branched chain fatty acid production. Microgels encapsulated with quercetin with or without KGM60 decreased Firmicutes while increased Bacteroidetes over 24 h of fermentation, at genus level promoted Bacteroides growth at 24 h and decreased the abundance of Negativibacillus, Ruminococcus_NK4A214, and Christensenellaceae R_7. Faecalibacterium and Collinsella levels were exclusively promoted by microgels encapsulating KGM60 with or without quercetin, highlighting prebiotic effect of KGM60. Only microgels co-encapsulating both KGM60 and quercetin enhanced Dialister while inhibited Lachnoclostridium, indicating synergism between KGM60 and quercetin. Our study indicates that co-encapsulating KGM60 and quercetin in alginate microgel is effective in modulating human gut microbiota and metabolites production potentially beneficial to gut health.

The Metabolic Footprint of Systemic Effects in the Blood Caused by Radiotherapy and Inflammatory Conditions: A Systematic Review.

Response to radiotherapy (RT) includes tissue toxicity, which may involve inflammatory reactions. We aimed to compare changes in metabolic patterns induced at the systemic level by radiation and inflammation itself. Patients treated with RT due to head and neck cancer and patients with inflammation-related diseases located in the corresponding anatomical regions were selected. PubMed and Web of Science databases were searched from 1 January 2000 to 10 August 2023. Twenty-five relevant studies where serum/plasma metabolic profiles were analyzed using different metabolomics approaches were identified. The studies showed different metabolic patterns of acute and chronic inflammatory diseases, yet changes in metabolites linked to the urea cycle and metabolism of arginine and proline were common features of both conditions. Although the reviewed reports showed only a few specific metabolites common for early RT response and inflammatory diseases, partly due to differences in metabolomics approaches, several common metabolic pathways linked to metabolites affected by radiation and inflammation were revealed. They included pathways involved in energy metabolism (e.g., metabolism of ketone bodies, mitochondrial electron transport chain, Warburg effect, citric acid cycle, urea cycle) and metabolism of certain amino acids (Arg, Pro, Gly, Ser, Met, Ala, Glu) and lipids (glycerolipids, branched-chain fatty acids). However, metabolites common for RT and inflammation-related diseases could show opposite patterns of changes. This could be exemplified by the lysophosphatidylcholine to phosphatidylcholine ratio (LPC/PC) that increased during chronic inflammation and decreased during the early phase of response to RT. One should be aware of dynamic metabolic changes during different phases of response to radiation, which involve increased levels of LPC in later phases. Hence, metabolomics studies that would address molecular features of both types of biological responses using comparable analytical and clinical approaches are needed to unravel the complexities of these phenomena, ultimately contributing to a deeper understanding of their impact on biological systems.

Evaluation of yeast-based additives, as an alternative to ionophores, on rumen fermentation of ruminant diets using an in vitro gas production system

IntroductionThe study aimed to assess yeast-based additives' effects, as monensin alternatives, on rumen fermentation parameters, greenhouse gas emissions, and ruminal kinetics of ruminant diets using an in vitro system. Three experiments were conducted, each individually evaluating escalating levels of three yeast-based additives.MethodsThree experiments were designed: Experiment 1 evaluated prebiotic blend 1—yeast culture [Saccharomyces cerevisiae (Scer)], beta-glucans, fructooligosaccharides, galactooligosaccharides, and mannanoligosaccharides; Experiment 2 investigated prebiotic blend 2—beta-glucan fractions and mannanoligosaccharides from Scer; Experiment 3 examined yeast cells—hydrolyzed, inactivated, and spray-dried yeast (Scer) cells. Uniform experimental design and procedures were employed across the three experiments. Each experiment had six treatments: monensin (Rumensin®, 25 mg/kg DM) as positive control, and yeast additive levels (0, 533, 1,067, 1,600, and 2,133 mg/kg on DM basis) added to ruminant diets (60% corn silage and 40% concentrate). An in vitro gas production (GP) system with 50 AnkomRF bottles assessed total GP (at 24 and 48 hours), kinetics, fermentation profiles, methane (CH4), and carbon dioxide (CO2) concentrations. Comparison with monensin utilized Dunnett’s test (5%). Yeast additive levels were analyzed for linear and quadratic responses.ResultsIn Experiment 1, the 1,600 mg/kg yeast additive had lower concentrations of propionate, isobutyrate, valerate, and branched-chain volatile fatty acids (BCVFAs), and a higher acetate concentration and acetate-to-propionate ratio than monensin. In Experiment 2, the 1,600 mg/kg yeast additive led to lower total VFA and isovalerate concentrations than monensin. Additionally, compared to the 1,067 mg/kg yeast additive, monensin showed lower isovalerate concentration and higher NH3-N concentration. In Experiment 3, the 533 mg/kg yeast additive resulted in lower valerate and BCVFA concentrations, and higher CH4 and CO2 concentrations than monensin. Monensin had lower total VFA, butyrate, and acetate-to-propionate ratio, and higher propionate concentration compared to the 2,133 mg/kg yeast additive.DiscussionCollectively, these findings suggest yeast-based additives could be monensin alternatives, enhancing animal nutrient utilization efficiency and contributing to improved livestock sustainability.

Effect of Lactobacillaceae Probiotics on Colonic Microbiota and Metabolite Production in Cystic Fibrosis: A Comparative In Vitro Study.

Cystic Fibrosis-related gut dysbiosis (CFRGD) has become a recognised complication in children with this condition, and current evidence remains insufficient to guide the selection of probiotic strains for supplementation treatments. The aim of this study was to characterise the effect of three probiotic strains on CFRGD by means of a dynamic in vitro simulation of the colonic fermentation (SHIME®). The configuration of the system included three bioreactors colonised with the faecal inoculum of a child with cystic fibrosis. For 20 days, each bioreactor was supplied daily with either Lacticaseibacillus rhamnosus GG (ATCC 53103 TM), Limosilactobacillus reuteri (DSM 17938) or Lactiplantibacillus plantarum (DSM 22266). The baseline microbiota was characterised by a high abundance of Prevotella, Faecalibacterium and Acidaminococcus genera. After 20 days of supplementation, L. rhamnosus and L. plantarum reduced Prevotella significantly, and the three strains led to increased Faecalibacterium and Bifidobacterium and decreased Acidaminococcus, with some of these changes being maintained 10 days after ceasing supplementation. The metabolic activity remained unaltered in terms of short-chain fatty acids, but branched-chain fatty acids showed a significant decrease, especially with L. plantarum. Additionally, ammonia decreased at 20 days of supplementation, and lactate continuously increased with the three strains. The effects on colonic microbiota of L. rhamnosus, L. reuteri or L. plantarum were established, including increased beneficial bacteria, such as Faecalibacterium, and beneficial metabolites such as lactate; and on the other hand, a reduction in pathogenic genera, including Prevotella or Acidaminococcus and branched-chain fatty acids, overall supported their use as probiotics in the context of CFRGD.

Comparative effects of medium-chain fatty acids or phytobiotics-based feed additives on performance, caecum microbiota, volatile fatty acid production and intestinal morphology of broilers.

Antibiotic growth promoters have been utilized in broiler nutrition to alleviate the negative effects of the pathogenic microbes to promote performance. However, after the prohibition of antibiotics because of the increasing disclosure related to public health issues, various products have been developed as alternatives. This study was carried out to determine the effects of medium-chain fatty acids (MCFAs) or phytobiotics (essential oils [EOs] and alkaloids [ALKs]), blended feed additives on the growth performance, jejunum histomorphology, and cecal microbiota of broiler chickens. A total of 765 male Ross 308 chicks were randomly distributed into 5 experimental groups, each having 9 replicates with 17 chicks. The experimental procedures were as follows: a control group without supplementation (T1); control group+ MCFAs and EOs blend (T2); control group+ different EOs blend (T3); control group+ ALK sanguinarine (T4); and control group+ EOs and ALK piperine mixture (T5). The results showed that, broilers fed with MCFAs blended with EOs had significantly greater body weight gain during overall period in comparision to the control and T3 groups. Further, only MCFAs blended with EOs group significantly improved jejnum morphology in comparison with the control group (p≤0.05). Besides, the MCFAs blended with EOs group significantly elevated propionate, acetate and butyrate concentration, and decreased the concentration of branch chain fatty acids in caecum (p≤0.05). The results indicated that, the combination of MCFAs and EOs seems to have improvement effects and could be preferred as an efficient feed additive in broiler production.

Heptanoic and medium branched-chain fatty acids as anaplerotic treatment for medium chain acyl-CoA dehydrogenase deficiency

Triheptanoin (triheptanoylglycerol) has shown value as anaplerotic therapy for patients with long chain fatty acid oxidation disorders but is contraindicated in medium-chain acyl-CoA dehydrogenase (MCAD) deficiency. In search for anaplerotic therapy for patients with MCAD deficiency, fibroblasts from three patients homozygous for the most common mutation, ACADMG985A/G985A, were treated with fatty acids hypothesized not to require MCAD for their metabolism, including heptanoic (C7; the active component of triheptanoin), 2,6-dimethylheptanoic (dMC7), 6-amino-2,4-dimethylheptanoic (AdMC7), or 4,8-dimethylnonanoic (dMC9) acids. Their effectiveness as anaplerotic fatty acids was assessed in live cells by monitoring changes in cellular oxygen consumption rate (OCR) and mitochondrial protein lysine succinylation, which reflects cellular succinyl-CoA levels, using immunofluorescence (IF) staining. Krebs cycle intermediates were also quantitated in these cells using targeted metabolomics. The four fatty acids induced positive changes in OCR parameters, consistent with their oxidative catalysis and utilization. Increases in cellular IF staining of succinylated lysines were observed, indicating that the fatty acids were effective sources of succinyl-CoA in the absence of media glucose, pyruvate, and lipids. The ability of MCAD deficient cells to metabolize C7 was confirmed by the ability of extracts to enzymatically utilize C7-CoA as substrate but not C8-CoA. To evaluate C7 therapeutic potential in vivo, Acadm-/- mice were treated with triheptanoin for seven days. Dose dependent increase in plasma levels of heptanoyl-, valeryl-, and propionylcarnitine indicated efficient metabolism of the medication. The pattern of the acylcarnitine profile paralleled resolution of liver pathology including reversing hepatic steatosis, increasing hepatic glycogen content, and increasing hepatocyte protein succinylation, all indicating improved energy homeostasis in the treated mice. These results provide the impetus to evaluate triheptanoin and the medium branched chain fatty acids as potential therapeutic agents for patients with MCAD deficiency.

Extrusion of lupines with or without addition of reducing sugars: Effects on the formation of Maillard reaction compounds, partition of nitrogen and Nε-carboxymethyl-lysine, and performance of dairy cows

The extrusion of leguminous seeds induces the formation of Maillard reaction compounds (MRC) as a product of protein advanced glycation and oxidation, which lowers protein degradability in the rumen. However, the quantitative relationship between the parameters of pretreatment (i.e., addition of reducing sugars) and extrusion, and the formation of MRC has not been established yet. Moreover, the fate of the main stable MRC, Nε-carboxymethyl-lysine (CML), in the excretory routes has never been investigated in ruminants. We aimed to test the effects of the temperature of extrusion of white lupines with or without addition of reducing sugars on the formation of MRC, crude protein (CP) degradability in the rumen, N use efficiency for milk production (milk N/N intake), and performance of dairy cows. Two experiments with a replicated 4 × 4 Latin square design were conducted simultaneously with 16 (3 rumen-cannulated) multiparous Holstein cows to measure indicators of ruminal CP degradability (ruminal NH3 concentration, branched-chain volatile fatty acids), metabolizable protein supply (plasma essential AA concentration), N use efficiency (N isotopic discrimination), and dairy performance. In parallel, apparent total-tract digestibility of dry matter, organic matter, neutral detergent fibers, N, total Lys and CML, and partition of N and CML were measured with 4 cows in both experiments. The diets consisted on a DM basis of 20% raw or extruded lupines and 80% basal mixed ration of corn silage, silage and hay from permanent grasslands, pelleted concentrate, and a vitaminized mineral mix. Expected output temperatures of lupine extrusion were 115°C, 135°C, and 150°C, without and with the addition of reducing sugars before extrusion. The extrusion numerically reduced the in vitro ruminal CP degradability of the lupines, and consequently increased the predicted supply of CP to the small intestine. Nitrogen balance and urinary N excretion did not differ among dietary treatments in either experiment. Milk yield and N use efficiency for milk production increased with extrusion of lupines at 150°C without addition of reducing sugars compared with raw lupines. Nitrogen isotopic discrimination between dietary and animal proteins (the difference between δ15N in plasma and δ15N in the diet) were lower with lupines extruded at 150°C without and with addition of reducing sugars. Regardless of sugar addition, milk true protein yield was not affected, but milk urea concentration and fat:protein ratio were lower with lupines extruded at 150°C than with raw lupines. In the CML partition study, we observed that on average 26% of the apparently digested CML was excreted in urine, and a much lower proportion (0.63% on average) of the apparently digested CML was secreted in milk, with no differences among dietary treatments. In conclusion, we showed that the extrusion of white lupines without or with addition of reducing sugars numerically reduced enzymatic CP degradability, with limited effects on N partition, but increased milk yield and N use efficiency at the highest temperature of extrusion without addition of reducing sugars.

Supplementing branched-chain volatile fatty acids in dual-flow cultures varying in dietary forage and corn oil concentrations. III: Protein metabolism and incorporation into bacterial protein

Some cellulolytic bacteria cannot transport branched-chain AA (BCAA) and do not express complete synthesis pathways, thus depending on cross-feeding for branched-chain volatile fatty acid (BCVFA) precursors for membrane lipids or for reductive carboxylation to BCAA. Our objective was to assess BCVFA uptake for BCAA synthesis in continuous cultures administered high forage (HF) and low forage (LF) diets without or with corn oil (CO). We hypothesized that BCVFA would be used for BCAA synthesis more in the HF than in LF diets. To help overcome bacterial inhibition by polyunsaturated fatty acids in CO, BCVFA usage for bacterial BCAA synthesis was hypothesized to decrease when CO was added to HF diets. The study was an incomplete block design with 8 dual-flow fermenters used in 4 periods with 8 treatments (n = 4) arranged as a 2 × 2 × 2 factorial. The factors were: HF or LF (67 or 33% forage, 33:67 alfalfa:orchardgrass pellets), without or with supplemental CO (3% of dry matter), and without or with 2.15 mmol/d (5 mg/d 13C) each of isovalerate, isobutyrate, and 2-methylbutyrate for one combined BCVFA treatment. The flow of bacterial BCAA increased by 10.7% by supplementing BCVFA and 9.14% with LF versus HF; similarly, dosing BCVFA versus without BCVFA increased BCAA by 1.98% in total bacterial AA, whereas LF increased BCAA by 1.92% versus HF. Additionally, BCVFA supplementation increased bacterial AA flow by 16.6% when supplemented in HF - CO and 12.4% in LF + CO diets, but not in the HF + CO (-1.5%) or LF - CO (+6.7%) diets (Diet × CO × BCVFA interaction). The recovery of 13C in bacterial AA flow was 31% lower with LF than with HF. Of the total 13C recovered in bacteria, 13.8, 17.3, and 30.2% were recovered in Val, Ile, and Leu, respectively; negligible 13C was recovered in other AA. When fermenters were dosed with BCVFA, nonbacterial and total effluent flows of AA, particularly of alanine and proline, suggest decreased peptidolysis. Increased ruminal outflow of bacterial AA, especially BCAA, but also nonbacterial AA could potentially support postabsorptive responses from BCVFA supplementation to dairy cattle.

Biotransformation of okara (soybean residue) through solid-state fermentation using probiotic Bacillus subtilis and Bacillus coagulans

Okara or soybean residue is a side-stream product of soybean processing generated in large quantities. However, okara is often underutilised as it is prone to spoilage and has a short shelf life. In this study, we explored the feasibility of biotransformation of okara through solid-state fermentation using probiotic Bacillus spp. to modify its nutritional value and flavour property. Two probiotic Bacillus species, namely Bacillus subtilis CU1 (CNCM I-2745) and R0179, and Bacillus coagulans IS-2 and 123 were firstly screened. Further analyses were performed on the B. subtilis R0179 and B. coagulans 123 fermented okara due to their better growth after 24 h (10.28 and 9.24 log colony forming units (CFU)/g respectively). Our results showed that B. subtilis R0179 converted a significant portion of okara insoluble dietary fibre into soluble dietary fibre after fermentation, but not B. coagulans 123. In addition, the samples fermented with B. subtilis R0179 showed higher amounts of maltose, hemicellulosic sugars as well as aromatic, branched-chain and hydrophobic amino acids. Furthermore, the B. subtilis R0179 transformed okara had greater amounts of acetoin, carboxylic acids, pyrazines and branched-chain fatty acids, which may contribute to characteristic natto-like and cheesy flavour. Overall, our study shed light on the possibility of using probiotic bacilli to transform okara with enhanced nutritional value and flavour property, which may increase okara usage in foods.

Comparing the effects of corn fermented protein with traditional distillers dried grains fed to healthy adult dogs on stool quality, nutrient digestibility, and palatability

Traditional distillers dried grains, co-products from the ethanol industry, can be utilized as sustainable ingredients for pet food. However, negative consumer perception prevents their widespread use. Corn fermented protein (CFP) is produced using post-fermentation separation technology, resulting in a high protein ingredient, which may increase consumer appeal compared to traditional distillers dried grains. The study objective was to compare the effect of CFP with that of traditional distillers dried grains on stool quality, nutrient digestibility, and palatability when fed to dogs. The control diet (CON) contained 15% soybean meal and the experimental diets contained 3.5% brewer’s dried yeast (BDY), 2.5% brewer’s dried yeast plus 17.5% distillers dried grains with solubles (BDY+DDGS), or 17.5% corn fermented protein (CFP). Experimental diets were fed to adult dogs (n = 12) in a 4 × 4 replicated Latin square design. Dogs were adapted to the diets for 9 days; this was followed by 5 days of total fecal collection. Titanium dioxide (0.4%) was added to all diets as an external marker to estimate digestibility. Data were analyzed using a mixed model in SAS, with treatment as a fixed effect and dog and period as random effects. Fecal output was greatest (p&amp;lt; 0.05) for dogs fed BDY+DDGS. The feces of dogs consuming CFP were firmer (p&amp;lt; 0.05) than those of dogs consuming CON and BDY+DDGS. Overall, nutrient digestibility was greatest (p&amp;lt; 0.05) for CON and BDY and lowest for BDY+DDGS, with that for CFP being intermediate. There were no differences (p &amp;gt; 0.05) in total short-chain or branched-chain fatty acid concentrations in the fresh fecal samples of dogs fed these dietary treatments. However, the percentage of propionate was higher (p&amp;lt; 0.05) in the fecal samples of dogs fed CON than in those of dogs fed BDY+DDGS, whereas the percentage of valerate was higher (p&amp;lt; 0.05) in the fecal samples of dogs fed CON than in those of dogs fed CFP. In the palatability evaluation, dogs had no preference when CON was compared with BDY or BDY+DDGS. However, dogs appeared to prefer CON over CFP. Overall, CFP resulted in improved stool quality and nutrient digestibility when compared with DDGS, which could increase consumer appeal for inclusion into pet food. The impact of CFP on palatability, however, warrants further investigation.

Individual and Group-Based Effects of In Vitro Fiber Interventions on the Fecal Microbiota.

The development of microbiome-targeted strategies is limited by individual differences in gut microbiome composition and metabolic responses to interventions. In vitro models that can replicate this variation allow us to conduct pre-clinical studies and assess efficacy. This study describes the exposure of 16 individual fecal microbiota samples to 5 different fibers using an in vitro system for the anaerobic cultivation of bacteria. The individual microbiota differed in composition and metabolite profiles (short-chain fatty acids and branched-chain fatty acids) after incubation with the fibers. Furthermore, microbiota composition after fiber incubation was significantly different between subjects with good intestinal health and subjects with Inflammatory Bowel Disease (IBD). α-diversity was differently affected by dietary fibers; for example, exposure to psyllium resulted in increased diversity in the healthy group and in decreased diversity in the IBD group. Instead, the functional metabolic profile did not differ between the two groups. Finally, the combination of all fibers, tested on the microbiota from IBD subjects, resulted in stronger overall effects on both microbiota composition and metabolite production compared to the single fibers. These results confirm that incubation with dietary fiber results in different compositional and functional effects on individual microbiota and that in vitro models represent successful tools for studying individual fiber effects.

Supplementing branched-chain volatile fatty acids in dual-flow cultures varying in dietary forage and corn oil concentrations. II: Biohydrogenation and incorporation into bacterial lipids

To maintain membrane homeostasis, ruminal bacteria synthesize branched-chain fatty acids (BCFA) or their derivatives (vinyl ethers) that are recovered during methylation procedures as branched-chain aldehydes (BCALD). Many strains of cellulolytic bacteria require 1 or more branched-chain volatile fatty acid (BCVFA). Therefore, the objective of this study was to investigate BCVFA incorporation into bacterial lipids under different dietary conditions. The study was an incomplete block design with 8 continuous culture fermenters used in 4 periods with treatments (n = 4) arranged as a 2 × 2 × 2 factorial. The factors were high (HF) or low forage (LF, 67 or 33% forage, 33:67 alfalfa:orchardgrass), without or with supplemental corn oil (CO; 3% dry matter, 1.5% linoleic fatty acid), and without or with 2.15 mmol/d (5 mg/d 13C each of isovalerate, isobutyrate, and 2-methylbutyrate). After methylation of bacterial pellets collected from each fermenter's effluent, fatty acids and fatty aldehydes were separated before analysis by gas chromatography and isotope ratio mass spectrometry. Supplementation of BCVFA did not influence biohydrogenation extent. Label was only recovered in branched-chain lipids. Lower forage inclusion decreased BCFA in bacterial fatty acid profile from 9.45% with HF to 7.06% with LF and decreased BCALD in bacterial aldehyde profile from 55.4% with HF to 51.4% with LF. Supplemental CO tended to decrease iso even-chain BCFA and decreased iso even-chain BCALD in their bacterial lipid profiles. The main 18:1 isomer was cis-9 18:1, which increased (P < 0.01) by 25% from CO (data not shown). Dose recovery in bacterial lipids was 43.3% lower with LF than HF. Supplemental CO decreased recovery in the HF diet but increased recovery with LF (diet × CO interaction). Recovery from anteiso odd-chain BCFA and BCALD was the greatest; therefore, 2-methylbutyrate was the BCVFA primer most used for branched-chain lipid synthesis. Recovery in iso odd-chain fatty acids (isovalerate as primer) was greater than label recovery in iso even-chain fatty acids (isobutyrate as primer). Fatty aldehydes were less than 6% of total bacterial lipids, but 26.0% of 13C recovered in lipids were recovered in BCALD because greater than 50% of aldehydes were branched-chain. Because BCFA and BCALD are important in the function and growth of bacteria, especially cellulolytics, BCVFA supplementation can support the rumen microbial consortium, increasing fiber degradation and efficiency of microbial protein synthesis.

Supplementing branched-chain volatile fatty acids in dual-flow cultures varying in dietary forage and corn oil concentrations. I: Digestibility, microbial protein, and prokaryotic community structure

Branched-chain amino acids are deaminated by amylolytic bacteria to branched-chain volatile fatty acids (BCVFA), which are growth factors for cellulolytic bacteria. Our objective was to determine the dietary conditions that would increase the uptake of BCVFA by rumen bacteria. We hypothesized that increased forage would increase cellulolytic bacterial abundance and incorporation of BCVFA into their structure. Supplemental polyunsaturated fatty acids, supplied via corn oil (CO), should inhibit cellulolytic bacteria growth, but we hypothesized that additional BCVFA would alleviate that inhibition. Further, supplemental BCVFA should increase neutral detergent fiber degradation and efficiency of bacterial protein synthesis more with the high forage and low polyunsaturated fatty acid dietary combination. The study was an incomplete block design with 8 dual-flow continuous cultures used in 4 periods with 8 treatments (n = 4 per treatment) arranged as a 2 × 2 × 2 factorial. The factors were: high forage (HF) or low forage (LF; 67 or 33%), without or with supplemental CO (3% dry matter), and without or with 2.15 mmol/d (which included 5 mg/d of 13C each of BCVFA isovalerate, isobutyrate, and 2-methylbutyrate). The isonitrogenous diets consisted of 33:67 alfalfa:orchardgrass pellet, and was replaced with a concentrate pellet that mainly consisted of ground corn, soybean meal, and soybean hulls for the LF diet. The main effect of supplementing BCVFA increased neutral detergent fiber (NDF) degradability by 7.6%, and CO increased NDF degradability only in LF diets. Supplemental BCVFA increased bacterial N by 1.5 g/kg organic matter truly degraded (6.6%) and 0.05 g/g truly degraded N (6.5%). The relative sequence abundance decreased with LF for Fibrobacter succinogenes, Ruminococcus flavefaciens, and genus Butyrivibrio compared with HF. Recovery of the total 13C dose in bacterial pellets decreased from 144 µg/ mg with HF to 98.9 µg/ mg with LF. Although isotope recovery in bacteria was greater with HF, BCVFA supplementation increased NDF degradability and efficiency of microbial protein synthesis under all dietary conditions. Therefore, supplemental BCVFA has potential to improve feed efficiency in dairy cows even with dietary conditions that might otherwise inhibit cellulolytic bacteria.