Effect of biochanin A on corn grain (Zea mays) fermentation by bovine rumen amylolytic bacteria.

AIMS: The objective was to determine the effect of biochanin A (BCA), an isoflavone produced by red clover (Trifolium pratense L.), on corn fermentation by rumen micro-organisms. METHODS AND RESULTS: When bovine rumen bacterial cell suspensions (n = 3) were incubated (24 h, 39°C) with ground corn, amylolytic bacteria including group D Gram-positive cocci (GPC; Streptococcus bovis; enterococci) proliferated, cellulolytic bacteria were inhibited, lactate accumulated and pH declined. Addition of BCA (30 µg ml-1 ) inhibited lactate production, and pH decline. BCA had no effect on total amylolytics, but increased lactobacilli and decreased GPC. The initial rate and total starch disappearance was decreased by BCA addition. BCA with added Strep. bovis HC5 supernatant (containing bacteriocins) inhibited the amylolytic bacteria tested (Strep. bovis JB1; Strep. bovis HC5; Lactobacillus reuteri, Selenemonas ruminatium) to a greater extent than either addition alone. BCA increased cellulolytics and dry matter digestibility of hay with corn starch. CONCLUSIONS: These results indicate that BCA mitigates changes associated with corn fermentation by bovine rumen bacteria ex vivo. SIGNIFICANCE AND IMPACT OF THE STUDY: BCA could serve as an effective mitigation strategy for rumen acidosis. Future research is needed to evaluate the effect of BCA on mitigating rumen acidosis in vivo.

Degradation of spent craft brewer's yeast by caprine rumen hyper ammonia-producing bacteria.

UNLABELLED: Spent yeast from craft beers often includes more hops (Humulus lupulus L.) secondary metabolites than traditional recipes. These compounds include α- and ß- acids, which are antimicrobial to the rumen hyper ammonia-producing bacteria (HAB) that are major contributors to amino acid degradation. The objective was to determine if the hops acids in spent craft brewer's yeast (CY; ~ 3·5 mg g(-1) hops acids) would protect it from degradation by caprine rumen bacteria and HAB when compared to a baker's yeast (BY; no hops acids). Cell suspensions were prepared by harvesting rumen fluid from fistulated goats, straining and differential centrifugation. The cells were re-suspended in media with BY or CY. After 24 h (39°C), HAB were enumerated and ammonia was measured. Fewer HAB and less ammonia was produced from CY than from BY. Pure culture experiments were conducted with Peptostreptococcus anaerobiusBG1 (caprine HAB). Ammonia production by BG1 from BY was greater than from CY. Ammonia production was greater when exogenous amino acids were included, but similar inhibition was observed in CY treatments. These results indicate that rumen micro-organisms deaminated the amino acids in CY to a lesser degree than BY. SIGNIFICANCE AND IMPACT OF THE STUDY: Spent brewer's yeast has long been included in ruminant diets as a protein supplement. However, modern craft beers often include more hops (Humulus lupulus L.) than traditional recipes. These compounds include α- and ß- acids, which are antimicrobial to the rumen hyper ammonia-producing bacteria (HAB) that are major contributors to amino acid degradation. This study demonstrated that hops acids in spent craft brewer's yeast protected protein from destruction by HABin vitro. These results suggest that the spent yeast from craft breweries, a source of beneficial hops secondary metabolites, could have value as rumen-protected protein.

Effect of starch source (corn, oats or wheat) and concentration on fermentation by equine faecal microbiota in vitro.

AIMS: The goal was to determine the effect of starch source (corn, oats and wheat) and concentration on: (i) total amylolytic bacteria, Group D Gram-positive cocci (GPC), lactobacilli and lactate-utilizing bacteria, and (ii) fermentation by equine microbiota. METHODS AND RESULTS: When faecal washed cell suspensions were incubated with any substrate amylolytics increased over time. However, at 24 h there were 10 and 1000-fold more amylolytics with corn than wheat or oats respectively. Predominant amylolytics isolated were Enterococcus faecalis (corn, wheat) and Streptococcus bovis (oats). GPC increased with any substrate, but decreased during stationary phase in oats only. Lactobacilli decreased during stationary phase with corn only. By 24 h, oats had more lactate-utilizers and lactobacilli and fewer GPC than corn and wheat. More gas was produced from oats or wheat than from corn. CONCLUSIONS: These results indicate that the growth of bacteria and fermentative capacity associated with starch metabolism is starch source dependent. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates a relationship between starch source and microbial changes independent of host digestion. However, future research is needed to evaluate the effect of starch source on the hindgut microbial community in vivo.

Inhibition of fructan-fermenting equine faecal bacteria and Streptococcus bovis by hops (Humulus lupulus L.) ß-acid.

AIMS: The goals of this study were to determine if ß-acid from hops (Humulus lupulus L.) could be used to control fructan fermentation by equine hindgut micro-organisms, and to verify the antimicrobial mode of action on Streptococcus bovis, which has been implicated in fructan fermentation, hindgut acidosis and pasture-associated laminitis (PAL) in the horse. METHODS AND RESULTS: Suspensions of uncultivated equine faecal micro-organisms produced fermentation acids when inulin (model fructan) was the substrate, but ß-acid (i.e. lupulone) concentrations ≥9 ppm inhibited lactate production and mitigated the decrease in pH. Inulin-fermenting Strep. bovis was isolated from the ß-acid-free suspensions after enrichment with inulin. The isolates were sensitive to ß-acid, which decreased the viable number of streptococci in faecal suspensions, as well as growth, lactate production and the intracellular potassium of Strep. bovis in pure culture. CONCLUSIONS: These results are consistent with the hypothesis that hops ß-acid prevented the growth of fructan-fermenting equine faecal bacteria, and that the mechanism of action was dissipation of the intracellular potassium of Strep. bovis. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial hindgut fermentation of grass fructans has been linked to PAL and other metabolic disorders in horses. Hops ß-acid is a potential phytochemical intervention to decrease the growth of bacteria responsible for PAL.

Ruminal tryptophan-utilizing bacteria degrade ergovaline from tall fescue seed extract.

The objectives of this study were to evaluate degradation of ergovaline in a tall fescue [ (Schreb.) Darbysh.] seed extract by rumen microbiota ex vivo and to identify specific bacteria capable of ergovaline degradation in vitro. Rumen cell suspensions were prepared by harvesting rumen fluid from fistulated wether goats ( = 3), straining, and differential centrifugation. Suspensions were dispensed into anaerobic tubes with added Trypticase with or without extract (∼10 µg kg ergovaline). Suspensions were incubated for 48 h at 39°C. Samples were collected at 0, 24, and 48 h for ergovaline analysis and enumeration of hyper-ammonia producing (HAB) and tryptophan-utilizing bacteria. Ergovaline values were analyzed by repeated measures using the mixed procedure of SAS. Enumeration data were log transformed for statistical analysis. When suspensions were incubated with extract, 11 to 15% of ergovaline disappearance was observed over 48 h ( = 0.02). After 24 h, suspensions with added extract had 10-fold less HAB than controls ( = 0.04), but treatments were similar by 48 h ( = 1.00). However, after 24 h and 48 h, suspensions with extract had 10-fold more tryptophan-utilizing bacteria ( < 0.01) that were later isolated and identified by their 16S RNA gene sequence as . The isolates and other known rumen pure cultures ( JB1, B159, HD4, B, F, MD1, SR) were evaluated for the ability to degrade ergovaline in vitro. Pure culture cell suspensions were incubated as described above and samples were taken at 0 and 48 h for ergovaline analysis. Data were analyzed using the ANOVA procedure of SAS. All HAB, including the isolates, tested degraded ergovaline (54 to 75%; < 0.05). B14 was also able to degrade ergovaline but to a lesser capacity (12%; < 0.05), but all other bacteria tested did not degrade ergovaline. The results of this study indicate which rumen bacteria may play an important role in ergovaline degradation and that microbiological strategies for controlling their activity could have ramifications for fescue toxicosis and other forms of ergotism in ruminants.

Phosphorus digestibility and phytate degradation by yearlings and mature horses.

Inorganic P is often added to growing horse diets because organic P, or phytate-P, is believed to have lower digestibility. If horses can efficiently digest organic P, then the need for inorganic P may be reduced. Much of the P in grain-based concentrates fed to growing horses is in the form of phytate-P. Little is known about the ability of growing horses to degrade phytate-P or whether horse age affects mineral digestion in horses. The objective of this study was to examine the effect of age on P, Ca, and Mg digestibility as well as phytate-P degradation. Four yearling geldings and 4 mature geldings were fed a diet of alfalfa cubes, timothy cubes, and a pelleted concentrate. The diet contained 0.28% total P and 17.4% of that P was in the phytate form. There was a 14-d diet adaptation period followed by a 4-d fecal collection period. Apparent total tract P digestibility was higher for yearlings than mature geldings ( = 0.036; 7.7 and -6.6% for yearlings and mature geldings, respectively). Phytate-P disappearance was 94.8% and did not differ between ages ( = 0.190). Apparent Ca digestibility was lower in mature geldings ( = 0.043), but apparent Mg digestibility did not differ between ages ( = 0.414). Phytate is broken down in the gastrointestinal tract, but the low P digestibilities suggest that either degradation occurs after the site of P absorption or liberated P is recycled back into the gastrointestinal tract. Yearlings can utilize organic P as well as mature horses; therefore, diets without inorganic P are acceptable for growing horses.