The objective of this clinical trial was to evaluate the effectiveness of probiotic, prebiotic, and synbiotic supplementation on average daily weight gain (ADG), duration of diarrhea, age at incidence of diarrhea, fecal shedding of Cryptosporidium oocysts, enteric pathogens, and the odds of pneumonia in preweaning dairy heifer calves on a commercial dairy. Feeding prebiotics and probiotics may improve health and production of calves. Hence, healthy Holstein heifer calves (n = 1,801) from a large California dairy were enrolled at 4 to 12 h of age and remained in this study until weaning at 60 d of age. Calves were block randomized to 1 of 4 treatments: (1) control, (2) yeast culture enriched with mannan-oligosaccharide (prebiotic), (3) Bacillus subtilis (probiotic), and (4) combination of both products (synbiotic), which were fed in milk twice daily from enrollment until weaning. Serum total protein at enrollment and body weight at 7, 42, and 56 d of age were measured. Fecal consistency was assessed daily for the entire preweaning period. A subgroup of 200 calves had fecal samples collected at 7, 14, 21, and 42 d for microbial culture and enumeration of Cryptosporidium oocysts by direct fluorescent antibody staining. Synbiotic-treated calves had 19 g increased ADG compared with control calves for overall ADG, from 7 to 56 d. From 42 to 56 d, prebiotic-treated calves had 85 g greater ADG and synbiotic-treated calves had 78 g greater ADG than control calves. There was no difference in duration of the first diarrhea episode, hazard of diarrhea, or odds of pneumonia per calf with treatment. Probiotic-treated calves had 100 times lower fecal shedding of Cryptosporidium oocysts at 14 d and prebiotic-treated calves had fewer Escherichia coli and pathogenic E. coli at 42 d compared with control calves. Although there were no effects on duration of diarrhea or pneumonia incidence, greater ADG in the late preweaning period may reflect treatment effects on enteric pathogens during the rearing process. The decreased shedding of Cryptosporidium should reduce infectious pressure, environmental contamination, and public health risks from Cryptosporidium. Our findings suggest ADG and potential health benefits for calves fed prebiotics, probiotics, and synbiotics and can help the dairy industry make informed decisions on the use of these products in dairy production.

Fresh forage crops have traditionally been preserved by fermentation, either based on the spontaneous fermentation by epiphytic lactic acid bacterial populations or driven by the addition of lactic acid bacteria inoculant cultures. Although this method of preservation is effective at retarding the spoilage and pathogenic bacteria it does not eliminate these organisms from the forage. Spore-forming and other bacteria are often able to survive the low pH and organic acid levels in the fermented forages and outgrow during feed-out and in the animal. The purpose of this research was to understand the levels and diversity of pathogens in silage systems as an important prerequisite to develop effective ways to control them. From February 2016 to December of 2019, 866 corn silage samples and 577 haylage samples were collected from 457 US dairy farms across 27 states to assess the pathogenic levels in silage. Results indicate the clostridia levels in the haylage samples averaged 9.4 x 103 cfu/g with a range of less than 10 cfu/g to 4.1 x 106 cfu/g. Corn silage had a significantly lower level of clostridia (p<0.05) averaging 2.3 x 103 cfu/g. Clostridium perfringens was the dominant species identified in both haylage and corn silage and accounted for more than 60% of all the clostridia isolates. The other major clostridia species identified were C. beijerinckii, Paraclostridium bifermentens and C. butyricum. E. coli levels were also determined in the haylage and corn silage samples. In haylage, the average E. coli level was 2.1 x 104 cfu/g and ranged from less than 10 cfu/g to 2.5 x 106 cfu/g and were significantly higher (p<0.05) than levels detected in corn silage (average of 5.4 x 101 cfu/g). Further work is underway to determine the source of these pathogens and methods of controlling through the fermentation process.

Rutin, a natural flavonol glycoside, elicits its diverse health-promoting effects from the bioactivities of quercetin, its aglycone. While widely distributed in the vegetables and fruits of human diet, rutin is either absent or inadequate in common animal feed ingredients. Rutin has been supplemented to dairy cows for performance enhancement, but its metabolic fate in vivo has not been determined. In this study, plasma, urine, and rumen fluid samples were collected before and after the intraruminal dosing of 100 mg/kg rutin to 4 Holsteins, and then characterized by both targeted and untargeted liquid chromatography-mass spectrometry (LC-MS)-based metabolomic analysis. In plasma and urine, 4-methylcatechol sulfate was identified as the most abundant metabolite of rutin, instead of quercetin and its flavonol metabolites, and its concentration was inversely correlated with the concentration of p-cresol sulfate. In rumen fluid, the formation of 3,4-dihydroxyphenylacetic acid (DHPAA) and 4-methylcatechol after rapid degradation of rutin and quercetin concurred with the decrease of p-cresol and the increase of its precursor, 4-hydroxyphenylacetic acid. Overall, the formation of 4-methylcatechol, a bioactive microbial metabolite, as the dominant bioavailable metabolite of rutin and quercetin, could contribute to their beneficial bioactivities in dairy cows, while the decrease of p-cresol, a microbial metabolite with negative biological and sensory properties, from the competitive inhibition between microbial metabolism of rutin and tyrosine, has the potential to reduce environmental impact of dairy operations and improve the health of dairy cattle.

The objective was to determine the effects of immunomodulatory feed ingredient during post-weaning on cortisol concentration and fecal microbial populations of beef heifers. Commercial Angus heifers (n = 72) from two AI sires were blocked (n = 9) by sire and BW, randomly assigned to one of two pens (4 heifers/pen) per block, then assigned to treatments. Heifers were fed twice daily from d 0 to 60 (gain 0.75 kg/day) and top-dressed once daily with either 72g of Celmanax (CEL) or corn germ (CON; corn germ meal) per pen. After 60 days, two heifers per pen (n = 32) were randomly selected for a transportation challenge. Fecal grab samples were collected on d 0 and 69 of treatment, hr -24, 0, 24 of the challenge and 7 d post-challenge. Serum samples were collected at h 0, 4, 8, and 12 of the challenge. Clostridia and E. coli were enumerated from fecal samples using selective media. Isolates (≤ five isolates from each media per sample) were genetically tested to determine if they were C. perfringens or pathogenic E. coli. Fecal samples were enriched for detection of Salmonella. Pen was the experimental unit and data was analyzed by ANOVA or repeated measures analysis. Following treatment, decreased (P ≤ 0.05) populations of total E. coli, Salmonella, and C. perfringens were observed in CEL heifers compared to CON heifers, whereas clostridia and pathogenic E. coli were not different (P > 0.05) between treatments. Transportation stress increased (P ≤ 0.05) populations of clostridia, C. perfringens, total E. coli, and Salmonella, but decreased (P = 0.0252) pathogenic E. coli counts. Cortisol concentrations were decreased (P < 0.05) in CEL heifers compared to CON heifers throughout the challenge. In summary, supplementation of Celmanax post-weaning altered microbial populations and cortisol concentrations were reduced during transportation in beef heifers.

The objective was to determine the effects of an immunomodulatory feed ingredient following weaning on cytokine expression and fecal microbial populations of heifers. Commercial Angus heifers (n = 72) were weaned (227 ± 7 d of age), blocked by BW (n = 9 blocks) and randomly assigned to one of 2 pens per block. Pens within weight block (4 heifers/pen) were then randomly assigned to treatments. Heifers were fed twice daily from d 0-60 (to gain 0.75kg/day) and top-dressed with either 18g/heifer/d of the immunomodulatory feed ingredient (Celmanax; Arm and Hammer Animal Nutrition, Princeton, NJ, USA; CEL) or corn-germ meal (CON). Blood samples were collected on d 0, 15, 30, 45, 60 and fecal grab samples on d 0 of the feeding trial. After d 60, two heifers per pen (n=32) were randomly selected for a transportation challenge. Serum samples were collected at h 0, 4, 8, 12 and fecal grab samples at h -24, 0, 24 and 7d post-challenge. Blood samples were analyzed for interferonγ (IFNγ), interleukin-8 (IL-8), and haptoglobin (HP) using commercially available ELISA kits and qRT-PCR for genes of interest associated with cytokine expression. Fecal samples were enumerated for Clostridia and E. coli using selective media (≤ 5 isolates from each media/sample), tested to determine if they were C. perfringens or pathogenic E. coli, and then enriched for detection of Salmonella. Data was analyzed via ANOVA. During the feeding trial, HP was reduced (P = 0.018) in CEL compared to CON at d 15, 45, and 60, while IFNγ and IL-8 did not differ (P &gt; 0.080) between treatments. All cytokines were decreased (P &lt; 0.001) in CEL compared to CON during the challenge. During the feeding trial, HP mRNA was increased (P = 0.045) in CEL compared to CON at d 30 and 60. Similarly, IFNγ mRNA was increased (P = 0.040) in CEL compared to CON, however, other genes of interest did not differ (P &gt; 0.172). Both C. perfringens and total E. coli counts were decreased (P = 0.036) in CEL compared to CON at 24h after the start of the transportation challenge. Clostridia and pathogenic E. coli counts did not differ (P = 0.941) between treatments. Total Clostridia and E. coli counts were increased (P &lt; 0.014) 24h post-challenge. All microbial populations, except pathogenic E. coli, observed decreased (P ≤ 0.009) counts from 24h to 7d post-challenge. Overall, Celmanax supplementation decreased circulating cytokines, and altered microbial populations and gene expression, thus, may serve a role in preparing animals to better cope with immunological challenges.