Effects of Aleurone Supplementation on Glucose-Insulin Metabolism and Gut Microbiome in Untrained Healthy Horses

Abstract

Aleurone, a layer of the bran fraction, is deemed to be responsible for the positive health effects associated with the consumption of whole-grain products. Studies on rodents, pigs, and humans report beneficial effects of aleurone in five main areas: the reduction of oxidative stress, immunomodulatory effects, modulation of energy management, digestive health, and the storage of vitamins and minerals. Our study is the first aleurone supplementation study performed in horses. The aim of this study was to investigate the effect of an increase in the dose levels of aleurone on the postprandial glucose-insulin metabolism and the gut microbiome in untrained healthy horses. Seven adult Standardbred horses were supplemented with four different dose levels of aleurone (50, 100, 200, and 400 g/day for 1 week) by using a Latin square model with a 1-week wash out in between doses. On day 7 of each supplementation week, postprandial blood glucose-insulin was measured and fecal samples were collected. 16S ribosomal RNA (rRNA) gene sequencing was performed and QIIME2 software was used for microbiome analysis. Microbial community function was assessed by using the predictive metagenome analysis tool Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) and using the Metacyc database of metabolic pathways. The relative abundancies of a pathway were analyzed by using analysis of composition of microbiomes (ANCOM) in R. There was a significant dose-dependent increase in the postprandial time to peak of glucose (p = 0.030), a significant delay in the time to peak of insulin (p = 0.025), and a significant decrease in both the insulin peak level (p = 0.049) and insulin area under the curve (AUC) (p = 0.019) with increasing dose levels of aleurone, with a consideration of 200 g being the lowest significant dose. Alpha diversity and beta diversity of the fecal microbiome showed no significant changes. Aleurone significantly decreased the relative abundance of the genera Roseburia, Shuttleworthia, Anaerostipes, Faecalibacter, and Succinovibrionaceae. The most pronounced changes in the relative abundance at phyla level were seen in Firmicutes and Verrucomicrobia (downregulation) and Bacteroidetes and Spirochaetes (upregulation). The PICRUSt analysis shows that aleurone induces a downregulation of the degradation of L-glutamate and taurine and an upregulation of the three consecutive pathways of the phospholipid membrane synthesis of the Archaea domain. The results of this study suggest a multimodal effect of aleurone on glucose-insulin metabolism, which is most likely to be caused by its effect on feed texture and subsequent digestive processing; and a synergistic effect of individual aleurone components on the glucose-insulin metabolism and microbiome composition and function.

The Wheat Aleurone Layer: Optimisation of Its Benefits and Application to Bakery Products

“…As an alternative to these biochemical markers, microscopy analyses can be performed to estimate the purity of the extracted fractions [110]. [12,119,120] ≈ 10.0-13.3 [7,12,121,122] -4.1 [80] 7.2-7.34 [123,124] 3.9 [125] 5.1 [126] MOISTURE (g/100 g) -8.0 [7] -5.4 [80,82] 8.14 [123] 6.7-8.98 [125,127] 7.0 [126] CARBOHYDRATES Arabinoxylans (g/100g) -24.3 [122] ----14.39 [126] A:X ratio 0.62 [128] 0.35-0.46 [7,122,128] ----β-glucans (g/100 g) 3.4 [128] 3.91-4.5 [122,128] ---1.7 [127] -Cellulose (g/100 g) 10.6 [128] 6.0 [128] -----…”

“….2-21.6 [129] -Starch (g/100 g) 1.9-5.8 [119,128] 2.2-5.8 [121,122,128] -36.5 [80,82] 2.5-12.75 [123,124] 33.46 [125] -Total Dietary Fibers (g/100 g) 39.7-60.0 [12,119,120,128] 39.7-49.2 [7,12,121,122,128] -15.4 [80,82] 43.36 [123] 27.90-37.3 [125,127] 44.3 [126] Soluble Dietary Fibers (g/100 g) 4.1 [120] ---3.07 [123] --Insoluble Dietary Fibers (g/100 g) 50.0 [120] ---40.15 [123] --…”

“…Total (g/100 g) 5.8-9.8 [119,120] 7.0-9.8 [7,121,122] -6.5 [80,82] 4.5-6.33 [123,124] 4.1-4.7 [129] -Calcium (Ca) (mg/100 g) 76.2 [120] -93 [10] ----Copper (Cu) (mg/100 g) ----1.35 [123] --Iron (Fe) (mg/100 g) 21.…”

“…3 [120] -26 [10] -13.93 [123] --Magnesium (Mg) (mg/100 g) 690-800 [12,120] 850 [12] 1030 [10] -770 [123] --Manganese (Mn) (mg/100 g) ----12.7 [123] --Sodium (Na) (mg/100 g) 1.7 [120] ------Phosphorus (P) (mg/100 g) 1900 [120] -2540 [10] -1857 [123] --Potassium (K) (mg/100 g) 1900 [120] -2250 [10] -1780 [123] --Zinc (Zn) (mg/100 g) 11.4 [120] -14.0 [10] -12.05 [123] --PHENELIC COMPOUNDS Total phenolic acids (mg/100 g) -----457 [127] -…”

“…Free (mg/100 g) --16.0 [121] 1.0-1.5 [121] ----- Bound (mg/100 g) Conjugated (mg/100 g) 0.60 [130] -0.99-1.0 [121,130] 0 [121] Sinapic acid -bound form (mg/100 g) 0.46 [130] 0.53 [130] -----Alkylresorcinols (mg/100 g) 1107 [130] 993.24 [130] ---138 [127] -Flavonoids (mg/100 g) 9.65 [130] 8.95 [130] -----Lignans (mg/100 g) 6300 [128] 4700 [128] -----Phytic acid (mg/100 g) 6900 [120] --2360 [80,82] ---PROTEINS Total (g/100 g) 13.3-18.0 [12,119,120] 21.0-22.2 [7,12,121,122] -23.6 [80,82] 15.2-17.46 [123,124] 16.5-21 [125,127] 20.2 [126] VITAMINS Total (mg/100 g) > 29.0 [120] 30.0 [121] -----Thiamin (B1) (mg/100 g) 0.87-1.6 [12,120] 1.1-1.4 [12,121] 1.4 [10] -1.26 [123] --Riboflavin (B2) (mg/100 g) 0.3 [120] 0.3 [121] 0.2 [10] -0.32 [123] --Niacin (B3) (mg/100 g) 24.0…”