Exogenous lactobacilli mitigate microbial changes associated with grain fermentation (corn, oats, and wheat) by equine fecal microflora ex vivo

PLoS One. 2017 Mar 30;12(3):e0174059. doi: 10.1371/journal.pone.0174059. eCollection 2017.

Abstract

Cereal grains are often included in equine diets. When starch intake exceeds foregut digestion starch will reach the hindgut, impacting microbial ecology. Probiotics (e.g., lactobacilli) are reported to mitigate GI dysbioses in other species. This study was conducted to determine the effect of exogenous lactobacilli on pH and the growth of amylolytic and lactate-utilizing bacteria. Feces were collected from 3 mature geldings fed grass hay with access to pasture. Fecal microbes were harvested by differential centrifugation, washed, and re-suspended in anaerobic media containing ground corn, wheat, or oats at 1.6% (w/v) starch and one of five treatments: Control (substrate only), L. acidophilus, L. buchneri, L. reuteri, or an equal mixture of all three (107 cells/mL, final concentration). After 24 h of incubation (37°C, 160 rpm), samples were collected for pH and enumerations of total amylolytics, Group D Gram-positive cocci (GPC; Enterococci, Streptococci), lactobacilli, and lactate-utilizing bacteria. Enumeration data were log transformed prior to ANOVA (SAS, v. 9.3). Lactobacilli inhibited pH decline in corn and wheat fermentations (P < 0.0001). Specifically, addition of either L. reuteri or L. acidophilus was most effective at mitigating pH decline with both corn and wheat fermentation, in which the greatest acidification occurred (P < 0.05). Exogenous lactobacilli decreased amylolytics, while increasing lactate-utilizers in corn and wheat fermentations (P < 0.0001). In oat fermentations, L. acidophilus and L. reuteri inhibited pH decline and increased lactate-utilizers while decreasing amylolytics (P < 0.0001). For all substrates, L. reuteri additions (regardless of viability) had the lowest number of GPC and the highest number of lactobacilli and lactate-utilizers (P < 0.05). There were no additive effects when lactobacilli were mixed. Exogenous lactobacilli decreased the initial (first 8 h) rate of starch catalysis when wheat was the substrate, but did not decrease total (24 h) starch utilization in any case. These results indicate that exogenous lactobacilli can impact the microbial community and pH of cereal grain fermentations by equine fecal microflora ex vivo. Additionally, dead (autoclaved) exogenous lactobacilli had similar effects as live lactobacilli on fermentation. This latter result indicates that the mechanism by which lactobacilli impact other amylolytic bacteria is not simple resource competition.

MeSH terms

  • Animal Nutritional Physiological Phenomena
  • Animals
  • Avena / chemistry
  • Digestion / physiology
  • Feces / chemistry
  • Feces / microbiology*
  • Fermentation / drug effects
  • Gram-Positive Cocci / chemistry
  • Gram-Positive Cocci / metabolism*
  • Horses / microbiology
  • Hydrogen-Ion Concentration
  • Lactobacillus / chemistry
  • Lactobacillus / metabolism*
  • Probiotics / chemistry
  • Probiotics / metabolism*
  • Starch / chemistry
  • Starch / metabolism
  • Triticum / chemistry
  • Zea mays / chemistry

Substances

  • Starch

Grants and funding

This work was supported by USDA-National Institute of Food and Agriculture (Hatch) and the WALTHAM BUCKEYE Equine Research Grant. MDF and GEA were supported by USDA-Agricultural Research Service National Program 215. Co-author Patricia Harris is employed by the funder, WALTHAM BUCKEYE. She had a role in study design and preparation of the manuscript, but no role in data collection or analysis. Her employer has a general interest in how probiotic lactobacilli work, but their products were not included in the study (neither were any other commercial products).