Invited review: "Probiotic" approaches to improving dairy production: Reassessing "magic foo-foo dust".

The gastrointestinal microbial consortium in dairy cattle is critical to determining the energetic status of the dairy cow from birth through her final lactation. The ruminant's microbial community can degrade a wide variety of feedstuffs, which can affect growth, as well as production rate and efficiency on the farm, but can also affect food safety, animal health, and environmental impacts of dairy production. Gut microbial diversity and density are powerful tools that can be harnessed to benefit both producers and consumers. The incentives in the United States to develop Alternatives to Antibiotics for use in food-animal production have been largely driven by the Veterinary Feed Directive and have led to an increased use of probiotic approaches to alter the gastrointestinal microbial community composition, resulting in improved heifer growth, milk production and efficiency, and animal health. However, the efficacy of direct-fed microbials or probiotics in dairy cattle has been highly variable due to specific microbial ecological factors within the host gut and its native microflora. Interactions (both synergistic and antagonistic) between the microbial ecosystem and the host animal physiology (including epithelial cells, immune system, hormones, enzyme activities, and epigenetics) are critical to understanding why some probiotics work but others do not. Increasing availability of next-generation sequencing approaches provides novel insights into how probiotic approaches change the microbial community composition in the gut that can potentially affect animal health (e.g., diarrhea or scours, gut integrity, foodborne pathogens), as well as animal performance (e.g., growth, reproduction, productivity) and fermentation parameters (e.g., pH, short-chain fatty acids, methane production, and microbial profiles) of cattle. However, it remains clear that all direct-fed microbials are not created equal and their efficacy remains highly variable and dependent on stage of production and farm environment. Collectively, data have demonstrated that probiotic effects are not limited to the simple mechanisms that have been traditionally hypothesized, but instead are part of a complex cascade of microbial ecological and host animal physiological effects that ultimately impact dairy production and profitability.

Dietary supplementation with live or autolyzed yeast: Effects on performance, nutrient digestibility, and ruminal fermentation in dairy cows.

This study was conducted to evaluate the effects of live or autolyzed yeast supplementation on dairy cow performance and ruminal fermentation. Two experiments were conducted to evaluate performance, feed sorting, total-tract apparent digestibility of nutrients, purine derivatives excretion, N utilization, ruminal fermentation, and the abundance of specific bacterial groups in the rumen. In experiment 1, 39 Holstein cows (171 ± 40 DIM and 32.6 ± 5.4 kg/d milk yield) were blocked according to parity, DIM, and milk yield and randomly assigned to the following treatments: control (CON); autolyzed yeast fed at 0.625 g/kg DM (AY; Levabon, DSM-Firmenich); or live yeast fed at 0.125 g/kg DM (LY; Vistacell, AB Vista). Cows were submitted to a 2-wk adaptation period followed by a 9-wk trial. In experiment 2, 8 ruminal cannulated Holstein cows (28.4 ± 4.0 kg/d milk yield and 216 ± 30 DIM), of which 4 were multiparous and 4 were primiparous, were blocked according to parity and enrolled into a 4 × 4 Latin square experiment with 21-d periods (the last 7 d for sampling). Cows within blocks were randomly assigned to treatment sequences: control (CON), LY (using the same product and dietary concentration as described in experiment 1), AY, or autolyzed yeast fed at 0.834 g/kg DM (AY2). In experiments 1 and 2, nutrient intake and total-tract apparent digestibility were not affected by treatments. Sorting for long feed particles (>19 mm) tended to be greater in cows fed yeast supplements than CON in experiment 1. Efficiency of N conversion into milk N was increased when feeding yeast supplements in experiment 1, and 3.5% FCM yield tended to be greater in cows fed yeast supplements than CON. Feed efficiency was increased when yeast supplements were fed to cows in relation to CON in experiment 1. In experiment 2, yield of FCM and fat were greater in cows fed yeast supplements compared with CON. Uric acid concentration and output in urine were increased when feeding yeast supplements when compared with CON. Neither ruminal pH nor total VFA were influenced by treatments. The current study did not reveal treatment differences in ruminal abundance of Anaerovibrio lipolytica, the genus Butyrivibrio, Fibrobacter succinogenes, Butyrivibrio proteoclasticus, or Streptococcus bovis. Yeast supplementation can increase feed efficiency without affecting nutrient intake and digestibility, ruminal VFA concentration, or ruminal abundance of specific bacterial groups. Supplementing live or autolyzed yeast, regardless of the dose, resulted in similar performance.

Effects of supplementing direct-fed microbials on health and growth of pre-weaning Gyr × Holstein dairy calves.

This study aimed to evaluate the effects of direct-fed microbials (DFM) on health and growth responses of pre-weaning Bos indicus × B. taurus (Gyr × Holstein) crossbred calves. Ninety newborn heifer calves [initial body weight (BW) 35 ± 4.0 kg] were used. At birth, calves were ranked by initial BW and parity of the dam and assigned to: 1) whole milk without DFM supplementation (CON; n = 30), 2) whole milk with the addition of 1.0 g/calf per day of a Bacillus-based DFM (BAC; n = 30), or 3) whole milk with the addition of 1.0 g/calf per day of BAC and 1.2 g/calf per day of Enterococcus faecium 669 (MIX; n = 30). Milk was fed individually during the study (77 d) and the BAC and MIX treatments were offered daily throughout the 77-d pre-weaning period. All calves were offered a starter supplement and corn silage starting on d 1 and 60 of age, respectively. Milk and starter supplement intake were evaluated daily, and BW was recorded on d 0 and at weaning (d 77). Diarrhea and pneumonia were assessed daily, and fecal samples were collected on d 0, 7, 14, 21, and at weaning (d 77) for assessment of the presence of bacterial and protozoal pathogens via qPCR. All data were analyzed using SAS (v. 9.4) with calf as the experimental unit and using single-df orthogonal contrasts (BAC + MIX vs. CON; BAC vs. MIX). Daily feeding of DFM, regardless of type, improved weaning BW. Odds ratio for occurrence of pneumonia was lower for DFM-supplemented calves, but occurrence of both did not differ between BAC and MIX calves. No Salmonella spp. or E. coli F41 were detected in any of the calves. The proportion of calves positive for E. coli F17 was greater for DFM calves on d 7 (92 and 96% vs. 81% for BAC, MIX, and CON, respectively), 21 (13 and 26% vs. 7% for BAC, MIX, and CON, respectively), and at weaning (48 and 35% vs. 22% for BAC, MIX, and CON, respectively). For C. difficile, more DFM calves were positive on d 7 (65 and 30% vs. 35% for BAC, MIX, and CON, respectively) and 14 (20 and 28% vs. 7% for BAC, MIX, and CON, respectively), but also greater for BAC vs. MIX on d 7. More CON calves were positive for C. perfringens on d 14 (14% vs. 3 and 8% for CON, BAC, and MIX, respectively) compared with DFM-fed calves. Incidence of calves positive for C. perfringens was greater in BAC vs. MIX on d 7 (50 vs. 18%), and greater for MIX vs. BAC at weaning (9 vs. 0%). For protozoa occurrence, a lower proportion of DFM calves were positive for Cryptosporidium spp. on d 7 (58 and 48% vs. 76% for BAC, MIX, and CON, respectively), but opposite results were observed on d 21 for Cryptosporidium spp. (3 and 11% vs. 0% for BAC, MIX, and CON, respectively) and Eimeria spp. on d 14 (7 and 8% vs. 0% for BAC, MIX, and CON, respectively) and 21 (50 and 59% vs. 38% for BAC, MIX, and CON, respectively). In summary, DFM feeding alleviated the occurrence of pneumonia, improved growth rates, while also modulating the prevalence of bacteria and protozoa in pre-weaning Gyr × Holstein calves.

Growth performance, nutrient utilization, rumen fermentation, blood biochemistry, and carcass traits of lambs fed Atriplex nummularia L. hay-based diet supplemented with yeast or bacterial direct- fed microbial.

Direct fed microbial may enhance the utilization of halophyte forages leading to improved animal growth and productivity. This study was conducted to evaluate Atriplex hay-based diet supplemented with yeast (Saccharomyces cerevisiae; SC) or bacteria (Bacillus subtilis and Lactobacillus casei; BAC) on lamb growth performance, digestibility, rumen fermentation, and carcass characteristics. Fifteen Barki lambs (90 ± 7 days of age and 18.6 ± 0.41 kg SE body weight; BW) were randomly assigned to three treatments for 120 days as follows: Control (basal diet without supplementation), SC and BAC diets, the basal diet supplemented with SC or BAC at 2 g/head/day, respectively. All lambs had similar dry matter (DM) intake, while lambs fed SC or BAC dies had higher (P < 0.05) total gain and average daily gain than those fed the control diet. Supplementation of SC or BC increased (P < 0.05) the digestibility of DM, organic matter, and acid detergent fiber, tended to decrease (P < 0.05) the urine N excretion and enhanced the N balance compared to the control. Ruminal pH, acetate, total volatile fatty acids concentrations, and bacterial protein were increased (P < 0.05), while creatinine and urea concentrations were decreased (P < 0.05) by both additives. Compared to other diets, the BAC diet reduced (P < 0.05) triglycerides, total lipids, kidney fat, and eye muscle fat. In conclusion, both additives resulted in similar positive growth performance and feed utilization, while only the BAC additive had a beneficial advantage in reducing the fat content of the carcass.

Effects of Bacillus subtilis PB6 supplementation on production, metabolism, inflammatory biomarkers, and gastrointestinal tract permeability in transition dairy cows.

Objectives were to evaluate the effects of Bacillus subtilis PB6 (BSP) on gastrointestinal tract permeability, metabolism, inflammation, and production parameters in periparturient Holstein cows. Multiparous cows (n = 48) were stratified by previous 305-d mature equivalent milk yield and parity and assigned to 1 of 2 top-dressed dietary treatments 21 d before expected calving through 63 DIM: (1) control (CON; 13 g/d calcium carbonate; n = 24) or (2) BSP (13 g/d BSP; CLOSTAT, Kemin Industries, Des Moines, IA; n = 24). Gastrointestinal tract permeability was evaluated in vivo using the oral paracellular marker chromium (Cr)-EDTA. Effects of treatment, time, and treatment × time were assessed using PROC MIXED of SAS version 9.4 (SAS Institute Inc.). Prepartum dry matter intake (DMI) was unaffected by treatment; however, BSP supplementation decreased postpartum DMI relative to CON (0.7 kg). Milk yield, energy-corrected milk (ECM), fat-corrected milk (FCM), and solids-corrected milk (SCM) increased in BSP cows compared with CON (1.6, 1.8, 1.6, and 1.5 kg, respectively). Decreased DMI and increased production collectively improved feed efficiency of milk yield, ECM, FCM, and SCM for BSP cows (6, 5, 5, and 5%, respectively). No treatment differences were observed for concentrations of milk fat, protein, total solids, somatic cell count, somatic cell score, body weight, or body condition score. Milk urea nitrogen concentrations decreased (5%), whereas milk protein and lactose yield increased (5 and 2%, respectively) with BSP supplementation. Prepartum fecal pH did not differ among treatments; conversely, postpartum fecal pH was increased with BSP supplementation (0.09 pH units). Prepartum fecal dry matter percentage, starch, acetic acid, propionic acid, butyric acid, and ethanol did not differ among treatments. Postpartum concentrations of the aforementioned fecal parameters were also unaffected by treatment, but fecal propionic acid concentration was decreased (24%) in BSP cows relative to CON. Circulating glucose, nonesterified fatty acids, l-lactate, and insulin were similar between treatments both pre- and postpartum. Prepartum ß-hydroxybutyrate (BHB) did not differ between treatments, but postpartum BSP supplementation decreased (21%) circulating BHB relative to CON. Regardless of treatment, inflammatory markers (serum amyloid A and haptoglobin) peaked immediately following parturition and progressively decreased with time, but this pattern was not influenced by treatment. Postpartum lipopolysaccharide binding protein tended to be decreased on d 3 in BSP relative to CON cows (19%). Neither treatment nor time affected Cr-EDTA area under the curve. In summary, supplementing BSP had no detectable effects prepartum, but increased key postpartum production parameters. Bacillus subtilis PB6 consistently increased postpartum fecal pH and decreased fecal propionate concentrations but did not appear to have an effect on gastrointestinal tract permeability.

Characterization of controlled trials on probiotic supplementation to dairy calves: A scoping review.

The objective of this scoping review was to identify, describe, and characterize the literature on probiotic supplementation in dairy calves. Eligible studies were nonrandomized, quasi-randomized and randomized controlled trials in English, Spanish, or Portuguese that evaluated the effect of probiotic supplementation on growth and health of dairy calves. The search strategies were based on a modification of the PICO (Population, Intervention, Comparator, Outcome) framework and used synonyms and words related to "dairy calves" (population), "probiotics" (intervention), and "growth and health measurements" (outcomes). No restrictions for publication year or language were applied. Searches were conducted in Biosis, CAB Abstracts, Medline, Scopus, and the Dissertations and Theses Database. In total, the search identified 4,467 records, of which 103 studies (110 controlled trials) met the inclusion criteria. The studies were published between 1980 and 2021 and originated from 28 countries. Trials were randomized (80.0%), nonrandomized (16.4%), and quasi-randomized (3.6%), ranging in sample size from 5 to 1,801 dairy calves (mode = 24; average = 64). Enrolled calves were frequently Holstein (74.5%), males (43.6%), and younger than 15 d at the beginning of probiotic supplementation (71.8%). Often, trials were conducted in research facilities (47.3%). Trials evaluated probiotics with single or multiple species of the same genus: Lactobacillus (26.4%), Saccharomyces (15.4%), Bacillus (10.0%), Enterococcus (3.6%), or multiple species of various genera (31.8%). Eight trials did not report the probiotic species used. Lactobacillus acidophilus and Enterococcus faecium were the species most supplemented to calves. The duration of probiotic supplementation ranged from 1 to 462 d (mode = 56; average = 50). In trials with a constant dose, it ranged from 4.0 × 106 to 3.7 × 1011 cfu/calf per day. Most probiotics were administered mixed solely into feed (88.5%; whole milk, milk replacer, starter, or total mixed ration) and less frequently orally as a drench or oral paste (7.9%). Most trials evaluated weight gain (88.2%) as a growth indicator and fecal consistency score (64.5%) as a health indicator. Our scoping review summarizes the breadth of controlled trials evaluating probiotic supplementation in dairy calves. Differences in intervention design (mode of probiotic administration, dose, and duration of probiotic supplementation) and outcomes evaluation (type and methods) justify future efforts toward standardized guidelines in clinical trials.

Effects of supplementing a direct-fed microbial containing Enterococcus faecium 669 on performance, health, and metabolic responses of preweaning Holstein dairy calves.

This study aimed to evaluate the effects of Enterococcus faecium 669 supplementation on performance, health, parasitological, microbiological, and hematological responses of preweaning dairy calves. Forty-two newborn Holstein female calves (initial body weight [BW] 44 ± 4.5 kg) were used in the present study. At birth, calves were ranked by initial BW and assigned to 1 of 2 treatment groups: (1) whole milk (CON; n = 21) and (2) whole milk with the addition of direct-fed microbial Ent. faecium 669 (DFM; n = 21). During the entire experimental period (63 d), DFM was daily-fed at a rate of 2.5 × 109 cfu/head. All calves were offered a mixture of a starter feed and wheat straw for ad libitum consumption. Supplement intake was evaluated daily, whereas calves were weighed on a weekly basis from d 0 to weaning (d 63). Diarrhea was assessed once a day, and fecal and blood samples were collected for microbiological, parasitological, and hematological responses. All data were analyzed with SAS using calf as the experimental unit. A treatment × week interaction was observed for BW, as DFM-supplemented calves were heavier than CON cohorts on d 56 (+ 4.7 kg) and at weaning on d 63 (+ 4.8 kg). A similar interaction was observed for average daily gain (ADG) and dry matter intake (DMI), with greater ADG for DFM-supplemented calves from d 35 to 42, greater ADG and DMI from d 49 to 56, and greater DMI from d 56 to weaning. Moreover, diarrhea occurrence tended to be lower, whereas rectal temperature was 0.2°C lower for DFM-supplemented calves. Treatment × day interactions were observed for the occurrence and counts of Eimeria spp., as DFM-supplemented calves tended to have a reduced number of positive observations on d 42 of the study versus CON, and a significant reduction in positive animals from d 21 to 42 was observed in the DFM group but not in CON calves. For Cryptosporidium spp., no treatment effects were observed on overall occurrence (%), but DFM-supplemented calves had a greater count of oocyst per gram versus CON. No treatment × day interaction or main treatment effects were observed for any of the blood variables analyzed herein, exception being monocytes concentration. In summary, preweaning Ent. faecium 669 supplementation improved performance, diarrhea occurrence, and reduced the number of calves positively-detected for Eimeria spp.

Bacillus-Based Direct-Fed Microbial Reduces the Pathogenic Synergy of a Coinfection with Salmonella enterica Serovar Choleraesuis and Porcine Reproductive and Respiratory Syndrome Virus.

Viral respiratory infections predispose lungs to bacterial coinfections causing a worse outcome than either infection alone. Porcine reproductive and respiratory syndrome virus (PRRSV) causes pneumonia in pigs and is often associated with bacterial coinfections. We examined the impact of providing weanling pigs a Bacillus-based direct-fed microbial (DFM) on the syndrome resulting from infection with either Salmonella enterica serotype Choleraesuis alone, or in combination with PRRSV. Nine days after the bacterial challenge, Salmonella was isolated from ileocecal lymph nodes of all challenged pigs regardless of DFM treatment. Compared to the single bacterial challenge, the dual challenge with Salmonella and PRRSV resulted in a pathogenic synergy exhibited by a higher rate of Salmonella colonization in the lung and a more extensive and severe interstitial pneumonia. Provision of DFM to dually challenged pigs reduced the rate of lung colonization by Salmonella, eliminated or reduced the presence of PRRSV in the lung, and reduced the extent and severity of gross lung pathology. Dually challenged pigs that received DFM had increased concentrations of interleukin 1 (IL-1) and IL-8 in lung lavage fluids, accompanied by increased expression in their blood cells of nucleotide-binding oligomerization domain receptor 2 (NOD2) and triggering receptor expressed in myeloid cells 1 (TREM-1) molecules. These changes in pulmonary inflammatory cytokine production and increased expression of NOD2 and TREM-1 suggest that the DFM exerted a systemic modulating effect on innate immunity. These observations are consistent with the notion that tonic stimulation by gut-derived microbial products can poise innate immunity to fight infections in the respiratory tract.

Evaluating the effects of Lactobacillus animalis and Propionibacterium freudenreichii on performance and rumen and fecal measures in lactating dairy cows.

Two experiments evaluated the effect of supplementation with a bacterial direct-fed microbial on performance and apparent total-tract nutrient digestion of dairy cows. In experiment 1, 30 multiparous cows (75 ± 32 d in milk) were randomly assigned to 1 of 2 treatments fed for 10 wk. All cows were fed a diet containing 23.8% starch. Treatments were top dressed to rations twice daily and consisted of a combination of Lactobacillus animalis (1 × 109 cfu/d) and Propionibacterium freudenreichii (2 × 109 cfu/d; LAPF) or carrier alone (CON). In experiment 2, 6 ruminally cannulated cows (123 ± 129 d in milk) were randomly assigned to a crossover design with two 6-wk periods. Cows received the same CON or LAPF treatment as in experiment 1. Cows were fed the same 23.8% starch diet as experiment 1 during wk 1 through 5 of each period, and then cows were abruptly switched to a 31.1% starch diet for wk 6. For both experiments, intake and milk yield were measured daily, and milk samples were collected weekly. In experiment 1, fecal grab samples were collected every 6 h on d 7 of experimental wk 1, 2, 4, 6, 8, and 10. Fecal consistency was scored, and fecal starch was measured in daily composite samples. Fecal composites from a subset of 7 cows per treatment were used to measure apparent total-tract nutrient digestion. In experiment 2, rumen pH was continuously recorded during wk 5 and 6. On d 7 of wk 5 (the final day of feeding the 23.8% starch ration), d 1 of wk 6 (the day of diet transition), and d 7 of wk 6 (the final day of feeding the 31.1% starch ration), rumen in situ digestion was determined. Samples of rumen fluid and feces were collected every 6 h on those days for measurement of fecal starch (composited by cow within day), rumen volatile fatty acids, and fecal pH. Rumen and fecal samples were collected at one time point on those days for microbiota assessment. In experiment 1, treatment did not affect intake, milk yield, milk composition, or fecal score. The LAPF treatment decreased fecal starch percentage and tended to increase starch digestion compared with CON, but the differences were very small (0.59 vs. 0.78% and 98.74 vs. 98.46%, respectively). Digestion of other nutrients was unaffected. In experiment 2, LAPF increased rumen pH following the abrupt switch to the high-starch diet, but milk yield was lower for LAPF compared with CON (35.7 vs. 33.2 kg/d). Contrary to the decrease in fecal starch with LAPF observed in experiment 1, fecal starch tended to be increased by LAPF following the abrupt ration change in experiment 2 (2.97 vs. 2.15%). Few effects of treatment on rumen and fecal microbial populations were detectable. Under the conditions used in our experiments, addition of the bacterial direct-fed microbials did not have a marked effect on animal performance, ruminal measures, or total-tract nutrient digestion.

Lactobacillus plantarum GB LP-1 as a direct-fed microbial for neonatal calves.

Direct-fed microbial feed additives with potential to enhance growth performance, gut health, and immunity have gained considerable popularity in neonatal calf production. Lactobacillus plantarum GB LP-1 (LP) produced by a proprietary fermentation process could be a viable direct-fed microbial feed for neonatal calves. The hypothesis was that feeding LP may ease transitioning from milk replacer (MR) to calf starter (CS) by improving gut health and appetite, while minimizing health challenges from pathogens and stress to improve growth performance. The experimental objective was to evaluate LP in an MR feeding program at 3 inclusion rates. Fifty-one 2- to 5-d-old Holstein bull calves were randomly assigned to 1 of 3 treatments using a randomized complete block design. Treatments were (1) Control (LP0): LP fed at 0 g/d; (2) LP4: LP fed at 4 g/d; and (3) LP8: LP fed at 8 g/d. Calves were fed MR at 0.57 kg/d for 14 d via bucket, which was increased to 0.85 kg/d until 35 d, and were then fed once daily at 0.425 kg/d with weaning after d 42 of the 56-d experiment. Calves were fed at 0630 and 1800 h in equal allotments, with access at all times to free-choice water and a pelleted CS with 25.5% crude protein. Calves demonstrated a linear growth response to increasing LP inclusion rate: calves fed LP8 gained more body weight (33.0, 36.9, and 37.7 kg for LP0, LP4, and LP8, respectively) than calves fed LP0, with calves fed LP4 being intermediate and similar. The 0-to-42-d (MR feeding phase) average daily gain (ADG; 562.9, 595.9, and 655.7 g/d) and 0-to-56-d ADG (588.6, 658.4, and 673.0 g/d) demonstrated linear responses, with calves fed LP8 having greater ADG than calves fed LP0, and calves fed LP4 being intermediate and similar. Total CS intake was similar among calves fed all treatments (66.3, 69.0, and 72.5 kg/56 d), which resulted in a quadratic response in feed efficiency (0.50, 0.53, and 0.52 kg of gain/kg of dry matter) for calves fed LP4 compared with calves fed LP0, with calves fed LP8 being intermediate and similar. Fecal scores improved linearly with increasing LP inclusion rate. These data demonstrate that feeding Lactobacillus plantarum GB LP-1 to neonatal calves improves gut health to increase growth performance at 4 and 8 g/d, while feed efficiency was greatest at 4 g/d.

Effectiveness of a Direct-Fed Microbial Product Containing Lactobacillus acidophilus and Lactobacillus casei in Reducing Fecal Shedding of Escherichia coli O157:H7 in Commercial Feedlot Cattle.

The objective of this study was to evaluate the effectiveness of a direct-fed microbial (DFM) product in reducing fecal shedding of Escherichia coli O157:H7 in finishing commercial feedlot cattle in Kansas (KS) and Nebraska (NE). Utilizing a randomized complete block design within the feedlot (KS, n = 1; NE, n = 1), cattle were randomly allocated to 20 pens, grouped in blocks of two based on allocation date, and then, within the block, randomly assigned to a treatment group (DFM or negative control). The DFM product was included in the diet at a targeted daily dose of 1 × 109 colony-forming units (CFU) of the Lactobacillus acidophilus and Lactobacillus casei combination per animal for at least 60 d before sampling. Feedlots were sampled for four consecutive weeks; weekly sampling consisted of collecting 20 pen floor fecal samples per pen. Fecal samples were subjected to culture-based methods for detection and isolation of E. coli O157, and positive samples were quantified using real-time polymerase chain reaction. Primary outcomes of interest were fecal prevalence of E. coli O157:H7 and E. coli O157 supershedding (≥104 CFU/g of feces) prevalence. Data for each feedlot were analyzed at the pen level using mixed models accounting for the study design features. Model-adjusted mean E. coli O157:H7 fecal prevalence estimates (standard error of the mean [SEM]) for DFM and control groups were 8.2% (SEM = 2.2%) and 9.9% (SEM = 2.5%) in KS and 14.6% (SEM = 2.8%) versus 14.3% (SEM = 2.6%) in NE; prevalence did not differ significantly between treatment groups at either site (KS, p = 0.51; NE, p = 0.92). Mean E. coli O157 supershedding prevalence estimates for DFM and control groups were 2.2% (SEM = 0.7%) versus 1.8% (SEM = 0.7%) in KS (p = 0.66) and 6.7% (SEM = 1.5%) versus 3.2% (SEM = 1.0%) in NE (p = 0.04). In conclusion, administering the DFM product in the finishing diet of feedlot cattle did not significantly reduce E. coli O157:H7 fecal prevalence or supershedding prevalence in study pens at either commercial feedlot.

Effect of feeding Azolla pinnata in combination with direct-fed microbial on broiler performance.

A study was conducted on broiler birds for 42 days to determine the effect of feeding azolla along with direct-fed microbial (DFM) on growth performance, nutrient utilization, biochemical parameters and carcass characteristics. A total of 360-day-old male broiler chicks were divided into 3 groups and 12 replicates with 10 chicks per replicate, according to complete randomized block design. One group was fed a diet containing 2.5% azolla (A) and another was fed a diet containing 2.5% azolla, with direct-fed microbial (A + DFM), viz. L. plantarum and L. rhamnosus, at the concentration of 13.4 × 108 and 1.52 × 108 CFU per mL, respectively, mixed in the daily quota of feed. The control group (CON) was fed a basal diet without azolla and DFM. Azolla fed alone and with DFM did not affect growth performance and feed conversion ratio (FCR) of birds during pre-starter and starter phase; however, significantly (P < 0.05) lower weight gain and higher FCR were observed during finisher period, resulting in an impaired overall FCR. Decreased (P < 0.05) plasma cholesterol and increased (P < 0.05) tibial calcium content were observed in birds fed azolla with DFM. The highest value (P < 0.05) of carcass characteristics like gizzard weight, breast muscle yield and lower meat pH were recorded in the birds fed azolla in combination with DFM, followed by birds fed azolla alone. In conclusion, azolla feeding at 2.5% level shows positive effect on breast muscle yield and gizzard weight of broiler which can be enhanced upon adding DFM to azolla-based diet.

Safety assessment of Weissella confusa - A direct-fed microbial candidate.

Weissella confusa is part of the lactic acid bacteria genera and a member of an autochthonous microbiota of humans and livestock. W. confusa has been proposed to have potential as a direct-fed microbial product, however, there is a lack of studies assessing its safety. A toxicological safety assessment of W. confusa was performed using a battery of in vitro, ex vivo and in vivo studies, testing for genotoxicity, skin and eye irritation and general toxicity. The bacterial reverse mutation (Ames) study did not reveal any genotoxicity in the presence and absence of metabolic activation (S9) at concentrations up to 5000 µg/mL. Moreover, an in vivo mammalian erythrocyte micronucleus study did not reveal any biologically relevant or statistically significant increases in the frequency of micronucleated polychromatic erythrocytes in mice, when tested at concentrations up to 2000 mg/kg body weight. W. confusa did not exert any skin irritation potential when tested in reconstructed skin membranes. When tested for eye irritation using an ex vivo model of isolated chicken eyes, mild irritation was observed. The 90-day sub-chronic oral toxicity (gavage) study was performed using Sprague Dawley rats at concentrations up to 92 × 108 cfu/kg body weight/day (nominal). The results showed that W. confusa were well tolerated, and no signs of toxicity were seen. The No Observed Adverse Effect Level (NOAEL) for both female and male animals was the highest concentration administered, 92 × 108 cfu/kg body weight/day (nominal). In conclusion, the toxicological studies performed confirmed W. confusa to be safe, making it a good candidate as a direct-fed microbial product.

Short communication: Production performance and nutrient digestibility of lactating dairy cows fed diets with and without addition of a live-yeast supplement.

The objective of this study was to evaluate the use of a live-yeast product when feeding relatively high-forage diets to high-producing cows in mid lactation. Eight primiparous [607 ± 43 kg of body weight (BW) and 130 ± 16 d in milk (DIM) at the beginning of the experiment] and 16 multiparous (706 ± 63 kg of BW and 137 ± 22 DIM at the beginning of the experiment) Holstein cows were blocked by parity and DIM, and randomly assigned to 1 of 2 diets (control vs. yeast) for a 12-wk period according to a randomized complete block design. The formulated diets contained 50.4% corn silage, 10.4% alfalfa hay, and 39.2% concentrate. The yeast diet was formulated to provide approximately 5.4 × 1011 cfu/d of Saccharomyces cerevisiae (BeneSacc; Global Nutritech Biotechnology LLC, Richmond, VA). Total-tract nutrient digestibility was estimated using 240-h undigested neutral detergent fiber (NDF) as an internal marker. Supplementing live yeast to lactating dairy cows did not affect dry matter intake (25.0 kg/d), milk yield (38.6 kg/d), milk fat concentration (4.78%), milk fat yield (1.83 kg/d), milk protein concentration (3.09%), milk protein yield (1.18 kg/d), milk lactose concentration (4.79%), milk lactose yield (1.84 kg/d), BW gain (-0.05 kg/d), or body condition score gain (0.16 units). The digestibility of dry matter was greater for the control treatment than for the yeast treatment (69.3 and 67.1%, respectively), but the digestibilities of crude protein (61.5%), NDF (40.5%), and starch (98.6%) were not affected by treatment. In conclusion, supplementation of live yeast did not affect production performance or nutrient digestibility of high-producing cows in mid lactation. The reasons for the lack of effect are not clear, but an evaluation of interactions between yeast and rumen buffer supplementation is warranted.

Effects of Saccharomyces cerevisiae-based direct-fed microbial and exogenous enzyme products on enteric methane emission and productivity in lactating dairy cows.

The objective of this experiment was to investigate the effects of a Saccharomyces cerevisiae-based direct-fed microbial product (SDM) and an exogenous enzyme product (ENZ) on enteric methane emission, milk yield and composition, total-tract digestibility of nutrients, ruminal fermentation, and nitrogen excretion and secretion in lactating dairy cows. Eighteen Holstein cows were used in a 3 × 3 Latin square design experiment with three 28-d periods. Treatments were (1) control (no additive), (2) 28 g of SDM/d per cow, or (3) 10 g of ENZ/d per cow. Treatments were top-dressed at the time of feeding. The basal diet consisted of (dry matter basis) 60% forage and 40% concentrates and contained 16.5% crude protein and 32.0% neutral detergent fiber. Treatments had no effect on enteric methane production, yield (methane per kg of dry matter intake, DMI), or intensity (methane per kg of energy-corrected milk yield). Carbon dioxide production was similar among treatments. Compared with control, SDM increased milk yield by 2 kg/d without affecting DMI or feed efficiency. Supplementation of the diet with ENZ did not affect DMI, milk yield, or feed efficiency. Concentrations and yields of milk fat, true protein, and lactose, and energy-corrected milk yield were not different among treatments. Neither SDM nor ENZ had an effect on total-tract digestibility of nutrients or nitrogen excretion and secretion. Concentration of total volatile fatty acids (VFA) in ruminal fluid was increased by both SDM and ENZ, and rumen pH was decreased by SDM compared with the control. At levels similar to the control DMI, the increased concentration of VFA in ruminal fluid of cows receiving SDM suggests an increased postruminal supply of energy and may partly explain the increased milk yield with that treatment. However, it is important to note that milk composition and energy-corrected milk yield were not affected by treatment.

Short communication: Effects of porcine plasma or combined sodium butyrate and Bacillus subtilis on growth and health of grain-fed veal calves.

The objective of this study was to evaluate the effects of incorporating 2 commonly used additives or spray-dried porcine plasma in calf milk replacer (CMR) on calf performance and health. Male Holstein calves (n = 158) transported from auction barns and local dairy farms were randomly assigned to receive 1 of 3 decoquinate-containing CMR for the first 49 d of the experiment: all milk protein and no additives (CONT); 15% of crude protein (CP) replaced with spray-dried porcine plasma, no additives (PLM); or all milk protein and an added combination of sodium butyrate (rate 1.4 kg of butyric acid/Mt) and Bacillus subtilis (1.28 million cfu/g of feed; BB). All milk replacers were formulated to contain 26% CP and 17% fat, 2.4% Lys, and 0.8% Met, and were bucket-fed at daily feeding rates of 520 g during wk 1 and 2, 650 g during wk 3, and 900 g during wk 4 and 5, in a total of 4, 5, and 6 L of solution, respectively. Calves were offered texturized calf starter (18% CP) upon arrival until wk 3 and transitioned to a corn and pellet ration with 2% straw (18.1% CP). No prophylactic administration of antibiotics occurred. All calves were gradually weaned over a 2-wk period. Calves were individually housed until weaned and then housed in groups of 5 in a mechanically ventilated facility in southwestern Ontario, Canada. Fecal scores, treatments administered (antibiotic or supportive therapy), and mortalities were recorded daily. Body weight was measured using a digital scale at arrival and at 14, 49, 56, and 78 d after arrival. No differences were found among the groups with respect to growth, feed efficiency, or incidence of diarrhea or respiratory infection treatment. Calves supplemented with BB had a greater hazard of mortality over the growing period compared with CONT. An interaction was found between the BB group and the level of total serum protein, with the BB group having a lower proportion of days with a fecal score of 3 when the calves had a higher total serum protein level. Calves fed PLM had a lower proportion of d with a fecal score of 3 relative to CONT but no difference in the proportion of d with a fecal score of 2 or higher. This study found that the addition of spray-dried plasma in CMR reduced diarrhea severity; however, supplementing BB was associated with a higher hazard of calf mortality and had a varying response on fecal score.

Literature-based safety assessment of an agriculture- and animal-associated microorganism: Weissella confusa.

Although Weissella confusa was established as a species over 25 years ago, it has been understudied until very recently. Several independent observations have driven the recent interest in this important microorganism. First, this Leuconostoc-like species of Lactic Acid Bacteria is associated with agricultural environments, many spontaneous food fermentations-especially carbohydrate-rich vegetable fermentations-and silage. Second, Weissella confusa are members of the autochthonous microbiota of healthy humans and livestock. Third, Weissella confusa-in a strain-specific fashion-are postulated to be good candidates for the development of novel direct-fed microbial products. Fourth, Weissella confusa-in a strain-specific fashion-have been described as opportunistic pathogens-especially in immunocompromised individuals. Last, a distantly related species (Weissella ceti) is the etiologic agent of weissellosis, a disease that affects farmed fish that are important for commercial aquaculture. The purpose of this literature-based safety assessment is to consolidate findings from primary research related to Weissella confusa and its natural associations with and effects on animals, humans, and their agricultural environments. Based on these assessments, it is reasonable to conclude that many Weissella confusa are safe for use in direct-fed microbial products for poultry.

Assessment of the potential of the multi-enzyme producer Bacillus amyloliquefaciens US573 as alternative feed additive.

BACKGROUND: Recently, probiotics have increasingly been used as feed additives in poultry diets as an alternative to antibiotic growth promoters fostering resistance development. RESULTS: This study was aimed at assessing the potential of Bacillus amyloliquefaciens US573 as a direct-fed microbial. The US573 strain was found to be free of harmful enzymatic activities and sensitive to antibiotics. In addition, it showed a good acid and bovine bile tolerance, high adhesion efficacy to chicken enterocytes, and an ability to form biofilms, which may favor its survival and persistence in the animal gastrointestinal tract. Moreover, besides the previously described extremely salt-tolerant and highly thermostable phytase, the US573 strain secretes xylanase, ß-glucanase and amylase activities useful in neutralizing antinutritional factors and maximizing the absorption of nutrients. The secretion of such enzymes may be responsible for the good performance of the US573 isolate in the digestibility of wheat in vitro. Indeed, using the vegetative cells, a yield of wheat dry matter digestibility of approximately 48% was achieved, which is slightly lower than the commercial feed additive Rovabio used as a reference (56.73% digestibility). CONCLUSION: The obtained results illustrate the potential of US573 strain as a promising direct-fed microbial candidate for application in the poultry industry. © 2017 Society of Chemical Industry.

Effects of bacterial direct-fed microbials on ruminal characteristics, methane emission, and milk fatty acid composition in cows fed high- or low-starch diets.

This study investigated the effects of bacterial direct-fed microbials (DFM) on ruminal fermentation and microbial characteristics, methane (CH4) emission, diet digestibility, and milk fatty acid (FA) composition in dairy cows fed diets formulated to induce different ruminal volatile fatty acid (VFA) profiles. Eight ruminally cannulated dairy cows were divided into 2 groups based on parity, days in milk, milk production, and body weight. Cows in each group were fed either a high-starch (38%, HS) or a low-starch (2%, LS) diet in a 55:45 forage-to-concentrate ratio on a dry matter (DM) basis. For each diet, cows were randomly assigned to 1 of 4 treatments in a Latin square design of (1) control (CON); (2) Propionibacterium P63 (P63); (3) P63 plus Lactobacillus plantarum 115 (P63+Lp); (4) P63 plus Lactobacillus rhamnosus 32 (P63+Lr). Strains of DFM were administered at 1010 cfu/d. Methane emission (using the sulfur hexafluoride tracer technique), total-tract digestibility, dry matter intake, and milk production and composition were quantified in wk 3. Ruminal fermentation and microbial characteristics were measured in wk 4. Data were analyzed using the mixed procedure of SAS (SAS Institute Inc., Cary, NC). The 2 diets induced different ruminal VFA profiles, with a greater proportion of propionate at the expense of acetate and butyrate for the HS diet. Greater concentrations of total bacteria and selected bacterial species of methanogenic Archaea were reported for the HS diet, whereas the protozoa concentration in HS decreased. For both diets, bacterial DFM supplementation raised ruminal pH (+0.18 pH units, on average) compared with CON. Irrespective of diet, P63+Lp and P63+Lr increased ruminal cellulase activity (3.8-fold, on average) compared with CON, but this effect was not associated with variations in ruminal microbial numbers. Irrespective of diet, no effect of bacterial DFM on ruminal VFA was observed. For the LS diet, supplementing cows with P63+Lr tended to decrease CH4 emission (26.5%, on average, when expressed per kilogram of milk or 4% fat-corrected milk). Only P63 supplementation to cows fed the HS diet affected the concentration of some milk FA, such as cis isomers of 18:1 and intermediates of ruminal biohydrogenation of polyunsaturated FA. Overall, bacterial DFM could be useful to stabilize ruminal pH. Their effects on CH4 production mitigation and milk FA profile depended on DFM strain and diet and should be confirmed under a greater variation of dietary conditions.

Effects of Direct-fed Microbial and Pine Cone Extract on Carcass Traits and Meat Quality of Hanwoo (Korean Native Cattle).

The carcass traits and meat quality of Hanwoo (Korean native cattle) whose diets were supplemented with direct-fed microbial (DFM) and pine cone extract (PCE) were evaluated. Twenty head of Hanwoo steers were divided equally into four groups and for a period of 6 months were given different diets: One group was fed a basal diet as control (CON), the other three groups were fed a basal diet supplemented with DFM-1%, DFM+PCE-1% and DFM+PEC-3%, respectively. DFM+PCE3% diet resulted the lowest carcass quality grade. The loins of DFM-1% contained higher moisture and lower fat than did the loins from the CON group. The crude protein content of DFM+PCE-3% group was significantly higher than that of the other groups. The water holding capacity and Warner-Bratzler shear force of the DFM+PCE-1% and 3% groups were lower than those of the CON and DFM-1% groups. The DFM-1% and 3% groups contained lower saturated fatty acid, higher unsaturated fatty acid, mono-unsaturated fatty acid, and poly-unsaturated fatty acid than did CON and DFM+PCE 1% group. Moreover, the n6:n3 ratios of DFM-1% and DFM+PCE-1% and 3% groups were slightly lower than that of the CON group. Thus we concluded that DFM and PCE supplementation resulted healthier Hanwoo beef with lower fat content and n6:n3 ratio.