Bacterial cultures, enzymes and yeast derived feed additives are often included in commercial dairy rations due to their effects on ruminal fermentation. However, the effects of these additives when fed together are not well understood. The objective of this study was to evaluate the changes in ruminal fermentation when a dairy ration is supplemented with combinations of bacterial probiotics, enzymes and yeast. Our hypotheses were that ruminal fermentation would be altered, indicated through changes in volatile fatty acid profile and nutrient digestibility, with inclusion of: (1) an additive, (2) yeast and (3) increasing additive doses. Treatments were randomly assigned to 8 fermenters in a replicated 4 × 4 Latin square with four 10 d experimental periods, consisting of 7 d for diet adaptation and 3 d for sample collection. Basal diets contained 52:48 forage:concentrate and fermenters were fed 106 g of dry matter per day divided equally between 2 feeding times. Treatments were: control (CTRL, without additives); bacterial culture/enzyme blend (EB, 1.7 mg per day); bacterial culture/enzyme blend with a blend of live yeast and yeast culture (EBY, 49.76 mg per day); and double dose of the EBY treatment (2X, 99.53 mg per day). The bacterial culture/enzyme blend contained five strains of probiotics (Lactobacillus animalis, Propionibacterium freudenreichii, Bacillus lichenformis, Bacillus subtillis, and Enterococcus faecium) and three enzymes (amylase, hemicellulase, and xylanase). On d 8-10, samples were collected for pH, redox, volatile fatty acids, lactate, ammonia N and digestibility measurements. Statistical analysis was performed using the GLIMMIX procedure of SAS. Repeated measures were used for pH, redox, VFA, NH3-N and lactate kinetics data. Orthogonal contrasts were used to test the effect of: (1) additives, ADD (CTRL vs EB, EBY and 2X); (2) yeast, YEAST (EB vs EBY and 2X); and (3) dose, DOSE (EBY vs 2X). No effects (p > 0.05) were observed for pH, redox, NH3-N, acetate, isobutyrate, valerate, total VFA, acetate:propionate, nutrient digestibility or N utilization. Within the 24h pool, the molar proportion of butyrate increased (p = 0.03) with the inclusion of additives when compared to the control while the molar proportion of propionate tended to decrease (p = 0.07). In conclusion, inclusion of bacterial cultures, enzymes and yeast to the diet increased butyrate concentration; but did not result in major changes in ruminal fermentation.

This report summarizes information on the usage of exogenous enzymes, such as their modes of action and effectiveness in lactating dairy cows. Written by S. L. Bennett and A. P. Faciola, and published by the UF/IFAS Department of Animal Sciences, March 2022.

This study aimed to evaluate the effects of Saccharomyces cerevisiae and Megasphaera elsdenii as direct fed microbials (DFM) in beef cattle finishing diets to alleviate acute ruminal lactic acidosis in vitro. A dual-flow continuous culture system was used. Treatments were a Control, no DFM; YM1, S. cerevisiae and M. elsdenii strain 1; YM2, S. cerevisiae and M. elsdenii strain 2; and YMM, S. cerevisiae and half of the doses of M. elsdenii strain 1 and strain 2. Each DFM dose had a concentration of 1 × 108 CFU/mL. Four experimental periods lasted 11 days each. For the non-acidotic days (day 1–8), diet contained 50:50 forage to concentrate ratio. For the challenge days (day 9–11), diet contained 10:90 forage to concentrate ratio. Acute ruminal acidosis was successfully established. No differences in pH, d-, l-, or total lactate were observed among treatments. Propionic acid increased in treatments containing DFM. For N metabolism, the YMM treatment decreased protein degradation and microbial protein synthesis. No treatment effects were observed on NH3–N concentration; however, efficiency of N utilization by ruminal bacteria was greater than 80% during the challenge period and NH3–N concentration was reduced to approximately 2 mg/dL as the challenge progressed.

Our objective was to evaluate the effects of bacteria (Lactobacillus animalis, Propionibacterium freudenreichii, Bacillus lichenformis, Bacillus subtilis, and Enterococcus faecium), enzymes (amylase, hemicellulose, and xylanase), and yeast as additives on the ruminal microbiome. We hypothesized that inclusion of bacteria, enzymes, and yeast would impact butyric bacterial populations. Eight fermenters were arranged in a duplicated 4 × 4 Latin square with the following treatments: (1) control without additives (CTRL); (2) bacterial culture and enzyme blend (EB); (3) bacterial culture and enzyme blend with a live yeast and yeast culture blend (EBY); and (4) double dose of bacterial culture and enzyme blend and the yeast products blend (2X). We conducted four fermentation periods of 10 d each, with the last 3 d for collection of samples. Overall, 64 solid and liquid samples were analyzed by amplification of the V4 region of bacterial 16S rRNA. Data were analyzed with R and SAS. The following orthogonal contrasts were used: (1) ADD—the control compared to all treatments with additives (CTRL vs. EB, EBY and 2X); (2) YEAST—treatment without yeast compared to those with yeast (EB vs. EBY and 2X); and (3) DOSE—the single dose of enzymes, bacteria and yeast compared to the doubled dose (EBY vs. 2X). Family Prevotellaceae was more abundant when additives were added (ADD). Additives (ADD) also increased relative abundance of Prevotellaceae Ga6A1 and YAB2003 in solid fraction, and of Prevotellaceae Ga6A1 and two members of Lachnospiracea family in liquid fraction. Yeast (YEAST) decreased relative abundance of Succinivibrionaceae UCG-001 and increased abundance of Ruminococcus and Prevotellaceae UCG-003 in solid fraction. Doubling the dose of enzymes and microbial additives (DOSE) decreased the abundance of Succiniclasticum in solid fraction and Selenomonadaceae in the liquid. Molar proportion of butyrate was highly correlated with abundance of Prevotellaceae Ga6A1 in solid (r = 0.68) and liquid fraction (r = 0.79), and with Unclassified Lachnospiraceae in liquid (r = 0.70). Our results demonstrate that YEAST decreases abundance of succinate synthesizing bacteria, while DOSE decreases abundance of bacteria that metabolize succinate into propionate. Combined bacteria, enzymes, and yeast increase the relative abundance of specific genera primarily within the Prevotellaceae family, which may explain the increase in butyrate molar proportion observed with ADD.

Supplemental sources of Mg can also aid in ruminal pH regulation due to their alkaline properties. Magnesium oxide (MgO) is the most common source of Mg for ruminants and can help controlling ruminal pH; however, the alkaline potential of other sources of Mg has not been evaluated. We aimed to evaluate the inclusion of calcium–magnesium carbonate (CaMg(CO3)2) and calcium–magnesium hydroxide (CaMg(OH)4) alone or in combination as supplemental sources of Mg in corn silage-based diets and its impact on ruminal microbial fermentation. We hypothesized that inclusion of CaMg(OH)4 would allow for ruminal fermentation conditions resulting in a greater pH compared to the inclusion of CaMg(CO3)2. Four treatments were defined by the supplemental source of Mg in the diet: 1) Control (100% MgO, plus sodium sesquicarbonate as a buffer); 2) CO3 [100% CaMg(CO3)2]; 3) OH [100% CaMg(OH)4]; and 4) CO3/OH [50% Mg from CaMg(CO3)2, 50% Mg from CaMg(OH)4]. Nutrient concentration was held constant across treatments (16% CP, 30% NDF, 1.66 Mcal NEl/kg, 0.67% Ca, and 0.21% Mg). Four fermenters were used in a 4 × 4 Latin square design with four periods of 10 d each. Samples were collected for analyses of nutrient digestibility, soluble Mg, VFA, and NH3, while pH was measured at 0, 1, 2, 4, 6, 8, and 10 h post morning feeding to estimate % time when pH was below 6 (pH-B6) and area under the pH curve for pH below 6.0 (pH-AUC). Bacteria pellets were harvested for 15N analysis and estimates of N metabolism. Treatment effects were analyzed with the mixed procedure of SAS, while effects of using either CaMg(CO3)2 or CaMg(OH)4 as Mg source in comparison to Control treatment were evaluated by orthogonal contrasts. Similar pH-related variables were observed for Control, OH, and CO3/OH treatments, which had smaller pH-AUC and pH-B6 than CO3 (P ≤ 0.01). Butyrate molar proportion was greater in Control and CO3/OH than in CO3 and OH (P = 0.04). Orthogonal contrasts showed lower flow of bacterial N (P = 0.04), lower butyrate molar proportion (P = 0.08) and greater pH-AUC (P = 0.05) for diets with CaMg(CO3)2 in comparison with the Control. Concentration of soluble Mg in ruminal fluid (P = 0.73) and nutrient digestibility (P ≥ 0.52) were similar across treatments. Under the conditions of this experiment, using CaMg(OH)4 alone or combined with CaMg(CO3)2 allowed for a less acidic ruminal fermentation pattern than a diet with only CaMg(CO3)2.

This exercise utilizes nightcrawler decomposition to guide students through the methods of science, including forming hypotheses, conducting an experiment, statistically analyzing data, and writing a lab report in the style of a scientific journal article. Additionally, students gain experience with dissection, anatomical terminology, and the biological process of animal decomposition. Students interested in forensic science will enjoy exploring the topic of decomposition. The materials for this experiment are inexpensive and easy to obtain, and several extension activities make this a versatile activity that can be used at many different educational levels.

Methane represents an energetic loss, and it has been proposed that its reduction may be associated with improved feed efficiency. Rumen methanogens produce most of the methane. Forty-nine crossbred heifers were individually fed a ration that consisted of 67.75% rolled corn, 20% wet distillers grain with solubles, 8% chopped alfalfa and 4.25% mineral/vitamin/rumensin mix for 84 d. Residual feed intake was the residual of the observed – predicted FI for the multivariate regression of FI on ADG and mid-metabolic BW for the population. Six heifers with the greatest RFI (1.75 ± 0.31 kg DM/d) and 6 with least RFI (-1.48 ± 0.31 kg DM/d) were selected. Rumen fluid was sampled from a tube passed through the mouth into the rumen. Rumen fluid was flash frozen in liquid N2 and stored at -80°C. After the first collection, heifers with greater RFI were pair fed to the less RFI heifers and rumen fluid was sampled 5 wk later. DNA was isolated from the rumen fluid and primer sets targeting total methanogens, Methanomicrobiales, Methanobacteriales, Methanosarcina, Methanobacterium, Methanobrevibacter ruminantium + Mbb. Cuticularis, and Methanobrevibacter smithii + Mbb. wolinii + Mbb. thaueri + Mbb. gottschalkii + Mbb. Woesei were amplified with real-time quantitative PCR (qPCR). Standard curves were generated to quantify copy number. There were no differences in copy number for the interaction of RFI classifications and sample period for all primer sets (P > 0.18). There were no differences in (P > 0.15) in RFI classification for all qPCR primer sets. There were fewer copies for the Methanobrevibacter ruminantium + Mbb. Cuticularis qPCR for samples collected in the first week compared to the 5 wk sample (P = 0.03); however, there were no difference for collection week for the other qPCR assays (P > 0.16). USDA is an equal opportunity provider and employer.

This report summarizes the information relating to the usage of direct-fed microbials, such as their modes of action and effectiveness in lactating dairy cows. Written by S. L. Bennett and A. P. Faciola, and published by the UF/IFAS Department of Animal Sciences, December 2021.