Effect of Barley Supplementation on the Fecal Microbiota, Caecal Biochemistry and Key

Effect of barley supplementation on the fecal microbiota, caecal biochemistry and key biomarkers of obesity and inflammation in obese db/db mice

SUPPLEMENTARY INFORMATION

Jose F. Garcia-Mazcorro1

1ResearchGroup Medical Eco-Biology, Faculty of Veterinary Medicine, Universidad Autónoma de Nuevo León, General Escobedo, Nuevo León, México

David A. Mills2

2Department of Food Science and Technology, University of California, Davis, California, USA Kevin Murphy3

3Department of Crop and Soil Sciences, Washington State University, Pullman, Washington, USA

Giuliana Noratto4,*,#

4School of Food Science, Washington State University, Pullman, Washington, USA

*Correspondence: Giuliana Noratto, Ph.D.

#Current address: Department of Nutrition and Food Science, Texas A&M University, College Station, TX, USA. Email:

Supplementary Figures

Figure S1 This figure shows all bacterial taxa that were found to be significantly increased in each treatment group accordingly to LEfSe analysis.

Supplementary Tables

Table S1Caloric intake and effect of barley consumption on biomarkers of obesity and organ weights

Lean / Obese control / Obese with Barley
Caloric intake* / 11.2a (2.1) / 16.6b (4.4) / 21.9c (8.7)
BMI (kg/m2) / 3.5a (1.0)
(n=11) / 5.8b (0.9) (n=10) / 6.4c(1.9)
(n=8)
Adiposity Index (AI) / 8.7a (9.0)
(n=9) / 26.6b (9.0) (n=9) / 26.2b (23.6)
(n=8)
Liver (g) / 1.3a (0.6) (n=10) / 2.8b (1.0) (n=10) / 2.5b (1.5)
(n=8)
Heart (mg) / 160a (0.1)
(n=11) / 140a (0.1) (n=10) / 130a(0.2)
(n=10)
Kidney (g) / 0.43a (0.1)
(n=11) / 0.45a (0.1) (n=9) / 0.43a (0.3)
(n=7)
Cecum tissue (mg) / 124.3a (143)
(n=11) / 65.3a,b (94) (n=10) / 259.2a (248)
(n=8)
Cecum content (mg) / 226.8a (246)
(n=11) / 119.3b (92) (n=10) / 367.5c (383)
(n=8)

Data are medians (range i.e. difference between the largest and smallest values). Statistical analysis was performed using the Kruskal-Wallis test in PAST. Multiple comparisons were adjusted by the Bonferroni method. Same superscripts indicate lack of statistical significant difference (P0.05). Please note the unequal sample size. * Caloric intake was calculated using eight values (from each treatment group), each representing the average from each week.

Table S2Effect of barley consumption on biomarkers of metabolic syndrome in plasma and liver cholesterol

Lean / Obese control / Obese with Barley
Fasting glucose (mmol/L) / 4.7a (2.5)
(n = 5) / 3.8a (1.2)
(n = 5) / 3.8a (2.6)
(n = 6)
Insulin (pg/mL) / 624.8a (3578)
(n=11) / 3528b (7197)
(n=10) / 1616c (1497)
(n=8)
Total-CHL (mg/dL) / 84a (43)
(n = 8) / 170b (108)
(n = 4) / 140b (73)
(n = 6)
LDL-CHL (mg/dL) / 43a (3)
(n = 8) / 83b (10)
(n = 4) / 71b (8)
(n = 6)
HDL-CHL (mg/dL) / 45a (35)
(n = 8) / 87a (95)
(n = 4) / 55a (79)
(n = 6)
Triglycerides (mg/dL) / 59a (82)
(n = 9) / 34b (24)
(n = 7) / 34b (32)
(n = 7)
Total-CHL in liver (mg/g liver) / 5.2a (6.5)
(n=9) / 13.7b (10.5)
(n=10) / 7.7a (4.7)
(n=5)

TableS3Effect of barley consumption on inflammatory markers in blood

Lean / Obese control / Obese with barley
Erythrocytes / ROS (RFU) / 1517a (858)
(n = 11) / 1899b (1268)
(n = 10) / 958c (653)
(n = 6)
Protein Carbonyls (nmol/mg protein) / 0.66a (0.3)
(n = 9) / 0.51a,b (1.1)
(n = 8) / 0.2b (0.3)
(n = 6)
Plasma / IL-6 (pg/mL) / 19a (179)
(n = 11) / 7.6b (178)
(n = 10) / 7.1b (59)
(n =8)
TBARS (µM MDA) / 27.7a (23)
(n = 7) / 102.8b (98)
(n = 8) / 64.6a,b (75)
(n = 5)
PAI-1(pg/mL) / 1462a (2053)
(n = 11) / 2320b (11605)
(n = 10) / 2423a,b (2297)
(n = 8)

Data are medians (range i.e. difference between the largest and smallest values). Statistical analysis was performed using the Kruskal-Wallis test in PAST. Multiple comparisons were adjusted by the Bonferroni method. Same superscripts indicate lack of statistical significant difference (P > 0.05). Please note the unequal sample size.Results from levels of ROS were expressed in arbitrary relative fluorescence units (RFU).

Table S4Effect of barley consumption on adipokines in plasma

Lean / Obese control / Obese with barley
Leptin (pg/mL) / 4038a (6095)
(n=11) / 16737b (32059)
(n=5*) / 15137b (34762)
(n=5**)
Resistin (pg/mL) / 1692a (1419)
(n=11) / 2074a,b (1696)
(n=10) / 1503a,c(1116)
(n=8)

Data are medians (range i.e. difference between the largest and smallest values). Statistical analysis was performed using the Kruskal-Wallis test in PAST. Multiple comparisons were adjusted by the Bonferroni method. Same superscripts indicate lack of statistical significant difference (P > 0.05). *Five samples for the obese group showed results for Leptin above the highest value standard curve and were therefore not included in this analysis. **Three samples for the barley group showed results for Leptin above the highest value standard curve and were therefore not included in this analysis.