A single intranasal dose of essential oil spray confers modulation of the nasopharyngeal microbiota and short-term inhibition of Mannheimia in feedlot cattle: a pilot study.

Five essential oils (EOs) were previously characterized in vitro and identified as candidate EOs for the development of an intranasal EO spray to mitigate bovine respiratory disease (BRD) pathogens. In the present study, these EOs were evaluated for their potential to (i) reduce BRD pathogens, (ii) modulate nasopharyngeal microbiota, and (iii) influence animal performance, feeding behavior and immune response when a single dose administered intranasally to feedlot cattle. Forty beef steer calves (7-8 months old, Initial body weight = 284 ± 5 kg [SE]) received either an intranasal EO spray (ajowan, thyme, fennel, cinnamon leaf, and citronella) or PBS (Control; n = 20/group) on day 0. Deep nasopharyngeal swabs were collected on days (d) -1, 1, 2, 7, 14, 28, and 42 and processed for 16S rRNA gene sequencing, qPCR, and culturing. Significant effects of EO on community structure (d1), microbial richness and diversity, relative abundance of some dominant phyla (d1, d2, and d14), and the overall interaction network structure of the nasopharyngeal microbiota were detected. The relative abundance of Mannheimia was lower in the EO calves (4.34%) than in Control calves (10.4%) on d2, and M. haemolytica prevalence on d7 as compared to control calves. Feed intake, average daily gain, feeding behavior, and blood cell counts were not affected by EO treatment. Overall, a single intranasal dose of EO spray resulted in moderate modulation of nasopharyngeal microbiota and short-term inhibition of Mannheimia while not influencing animal performance, feeding behavior or immune response. Our study, for the first time, shows the potential use of intranasal EO to mitigate BRD in feedlot cattle.

Impact of shade in beef feedyards on performance, ear temperature, and heat stress measures.

A 2-yr study (year 1: March to September 2017; year 2: February to August 2018) was conducted using crossbred steers (year 1: n = 1677; initial body weight [BW] = 372 kg, SD = 47; year 2: n = 1713; initial BW = 379 kg, SD = 10) in a commercial feedyard study in Eastern NE to determine the effects of shade on cattle performance, ear temperature, and cattle activity. Two treatments were evaluated using a randomized complete block design (n = 5 blocks based on arrival). Treatments were assigned randomly to pens and consisted of five pens without shade (NO SHADE) and five pens with shade (SHADE). Ear temperatures were collected throughout the trials using biometric sensing ear tags on a subset of cattle. Panting scores were collected using a 5 point scale determined visually based on the level of panting occurring on the same subset of steers a minimum of twice weekly from June 8 to August 21 in year 1 and May 29 to July 24 in year 2 by one trained individual each year. In year 1, no differences (P ≥ 0.24) were observed for growth performance or carcass characteristics. Dry matter intake (DMI) and average daily gain (ADG) were greater (P ≤ 0.04) for SHADE cattle in year 2. Over the entire feeding period in year 1, greater (P < 0.01) ear temperature was observed for NO SHADE cattle, but cattle movement was not different (P = 0.38) between treatments. When evaluating the entire feeding period in year 2, cattle movement and ear temperature were not different (P ≥ 0.80) between treatments. Cattle in the SHADE treatment had lower (P ≤ 0.04) panting scores in years 1 and 2. These data suggest that providing shade can lessen the negative influence of heat events on DMI and was an effective way to reduce heat stress in feedlot operations, but only impacted ADG if heat events were close to the cattle slaughter date.

This study was performed over two separate summers to evaluate natural exposure to heat and quantify the impact that providing shade has on stress measures, estimates of body temperature, and feedlot performance. In the first year, cattle were marketed after summer heat events and while ear temperature and panting scores were decreased for cattle in shaded pens, the effect was not significant enough to influence performance parameters. In the second year when cattle were marketed more immediately following heat impacts, cattle fed in shaded pens consumed more feed and gained more weight than cattle fed in pens without shade. In both years, cattle decreased dry matter intakes to offset normal heat events.

Feeding hempseed cake alters the bovine gut, respiratory and reproductive microbiota.

A growing number of studies have investigated the feasibility of utilizing hemp by-products as livestock feedstuffs; however, their impact on livestock microbiomes remains unexplored. Here, we evaluated the effects of feeding hempseed cake on the gastrointestinal, respiratory, and reproductive microbiota in beef heifers. Angus-crossbred heifers (19-months old, initial body weight = 494 ± 10 kg [SE]) were fed a corn-based finishing diet containing 20% hempseed cake as a substitute for 20% corn dried distillers' grains with solubles (DM basis; Control; n = 16/group) for 111 days until slaughter. Ruminal fluid and deep nasopharyngeal swabs (days 0, 7, 42, 70 and 98), and vaginal and uterine swabs (at slaughter) were collected, and the microbiota assessed using 16S rRNA gene sequencing. Diet affected the community structure of the ruminal (d 7-98; 0.06 ≤ R2 ≤ 0.12; P < 0.05), nasopharyngeal (d 98; R2 = 0.18; P < 0.001), and vaginal (R2 = 0.06; P < 0.01) microbiota. Heifers fed hempseed cake had increased microbial diversity in the rumen, reduced microbial richness in the vagina, and greater microbial diversity and richness in the uterus. In addition to the distinct microbial communities in the rumen, nasopharynx, vagina and uterus, we identified 28 core taxa that were shared (≥ 60% of all samples) across these sampling locations. Feeding hempseed cake appeared to alter the bovine gut, respiratory and reproductive microbiota. Our results suggest that future research aiming to evaluate the use of hemp by-products in livestock diet should consider their impact on animal microbiome and microbiome mediated animal health and reproductive efficiency. Our findings also highlight the need for research evaluating the impact of hemp-associated food and personal care products on the human microbiome.

Excretion and residue depletion of cannabinoids in beef cattle fed hempseed cake for 111 days.

Thirty-two crossbred heifers were fed either a control diet or 20% (dry matter basis) hempseed cake in a complete ration for 111 days; of the cattle fed hempseed cake, four each were harvested with 0, 1, 4, and 8-day withdrawal periods. Urine and plasma were collected during the feeding and withdrawal periods and liver, kidney, skeletal muscle, and adipose tissue were collected at harvest. Total cannabinoid (n = 10) concentration of hempseed cake averaged 11.3 ± 11.7 mg kg-1 across the feeding period with total cannabidiol and tetrahydrocannabinol (CBD/THC) concentrations of 1.3 ± 0.8 mg kg-1. Neutral cannabinoids (cannabinol [CBN], CBD/THC, and cannabidivarin [CBDV]) were not detected in plasma or urine, but CBD/THC was measured in adipose tissue at all withdrawal periods (6.3 ± 2.1 to 10.1 ± 2.5 ng g-1). In contrast, cannabinoid acids (cannabinolic acid [CBNA], cannabidiolic acid [CBDA]/tetrahydrocannabinolic acid [THCA], cannabichromenic acid [CBCA], and cannabidivarinic acid [CBDVA]) were sporadically detected (<15 ng mL-1) in plasma and urine of cattle fed hempseed cake. Cannabinoid acids were depleted from liver by withdrawal day 4, but could still be measured (<1 ng g-1) in kidney of some animals harvested on withdrawal day 8. Assessment of human exposures to CBD/THC residues through the consumption of beef fat from animals fed hempseed cake suggests that the probability of consuming the equivalent of an acute reference dose (ARfD) is remote, even with the use of a conservative reference dose ARfD (1 µg kg-1 body weight).

Effects of hempseed cake on ruminal fermentation parameters, nutrient digestibility, nutrient flow, and nitrogen balance in finishing steers.

Five ruminally and duodenally cannulated red angus steers (n = 5; initial body weight [BW] = 542 kg, SD = 40 kg) were used in a three-period Youden square design consisting of three 21-d periods, three treatments, and five steers (one or two steers per treatment within each period) to evaluate the effect of feeding hempseed cake on ruminal fermentation parameters, organic matter (OM) intake, total tract nutrient digestion, and nitrogen (N) balance in steers fed finishing diets. The control (CON) diet contained 75% dry-rolled corn, 20% corn silage, and 5% supplement (DM basis). The dried corn distillers grains plus solubles (DDGS) and hempseed cake (HEMP) diets contained 55% dry-rolled corn, 20% corn silage, 20% dried corn distillers grains plus solubles or hempseed cake, and 5% supplement (DM basis). Total ruminal volatile fatty acid concentration was greater (P < 0.01) in steers fed the HEMP diet than in steers fed the DDGS or CON diets. Ruminal fluid pH was not influenced (P = 0.93) by treatment. Organic matter intake tended (P = 0.07) to be greater and OM total tract digestibility was less (P = 0.03) in steers fed the HEMP diet compared with steers fed the DDGS or CON diets. Ruminal true and total tract apparent N digestibility was greater (P < 0.01) in steers fed the HEMP diet than steers fed the DDGS or CON diets. Duodenal flow of essential, nonessential, and total amino acids was not influenced (P ≥ 0.37) by dietary treatment, but the lack of response was likely because ruminally degradable protein (RDP) supply exceeded the RDP requirement. Steers fed the HEMP diet had greater (P < 0.01) N retention (g/d) than steers fed the DDGS diet, which was greater (P < 0.01) than steers fed the CON diet, suggesting that feeding hempseed cake improved utilization of N. Although inclusion of hempseed cake decreased total tract OM digestibility compared with dried corn distillers grains or corn, improvements in N utilization suggest that hempseed cake could be a useful alternative feed ingredient for finishing cattle diets.

This experiment evaluated the effects of dietary inclusion of hempseed cake or dried corn distillers grains plus solubles on ruminal fermentation parameters, organic matter (OM) intake, total tract nutrient digestion, and nitrogen (N) balance in steers fed finishing diets. Steers were assigned to one of three dietary treatments (no byproduct [CON], 20% dried corn distillers grains plus solubles [DDGS], or 20% hempseed cake [HEMP]; dry matter basis). Hempseed cake had greater acid detergent fiber concentrations, which resulted in greater acid detergent fiber flow to the small intestine and reduced total tract organic matter digestibility in steers fed the HEMP diet than in steers fed the DDGS or CON diets. Steers fed the HEMP diet had greater ruminal and total tract N digestibility, and greater ruminal ammonia concentrations than steers fed DDGS or CON diets, suggesting that the crude protein in hempseed cake is degraded to a greater extent in the rumen and total tract. Although inclusion of hempseed cake decreased total tract OM digestibility compared with dried corn distillers grains or no byproduct, the observed greater ruminal and total tract N digestibility suggest that it could be a useful alternative feed ingredient for finishing cattle diets.

Rapid quantification of cannabinoids in beef tissues and bodily fluids using direct-delivery electrospray ionization mass spectrometry.

Hempseed cake is a byproduct of hempseed oil extraction and is potentially a useful source of protein and fiber for use in ruminant diets. However, data are lacking on the appearance and/or clearance of cannabinoids in tissues of animals fed hempseed cake. To this end, a rapid method for quantifying cannabinol (CBN), cannabidiol (CBD), cannabinolic acid (CBNA), cannabidiolic acid (CBDA), cannabigerolic acid (CBGA), cannabichromenic acid (CBCA), cannabidivarin (CBDV), cannabidivarinic acid (CBDVA), tetrahydrocannabinol (THC) and tetrahydrocannabinolic acid (THCA) in cattle tissues, plasma, and urine was developed using rapid screen electrospray ionization mass spectrometry (RS-ESI-MS). Regression coefficients of matrix-matched standard curves ranged from 0.9946 to >0.9999 and analyte recoveries averaged from 90.2 ± 15.5 to 108.7 ± 18.7% across all compounds. Limits of detection and quantification ranged from 0.05 to 2.79 ng · mL-1 and 0.17 to 9.30 ng · mL-1, respectively, while the inter-day relative standard deviation ranged from 5.1 to 15.1%. Rapid screening electrospray ionization mass spectrometry (RS-ESI-MS) returned no false positives for any cannabinoid in plasma, urine, and tissue (liver, skeletal muscle) samples from 6 non-dosed control animals (n = 90 samples; of which 72 samples were plasma or urine and 18 samples were tissues). Across-animal cannabinoid concentrations measured in 32 plasma samples of cattle dosed with ground hemp were quantified by RS-ESI-MS; analytical results correlated well (r2 = 0.963) with independent LC-MS/MS analysis of the same samples.

Influence of hempseed cake inclusion on growth performance, carcass characteristics, feeding behavior, and blood parameters in finishing heifers.

As the hemp industry continues to develop in the United States, there is an increasing interest in feeding byproducts of industrial hemp production to livestock. A completely randomized design experiment using crossbred finishing heifers (initial body weight [BW] ±â€…SE = 494 ±â€…10 kg) was conducted to determine the effects of feeding hempseed cake in a corn-based finishing diet (10% forage) formulated to meet or exceed ruminally degradable and metabolizable protein requirements on growth performance, carcass characteristics, feeding behavior, and plasma parameters. Dietary treatments were the inclusion of 20% (dry matter [DM] basis) of dried corn distillers grains plus solubles (DDGS, n = 16) or hempseed cake (HEMP, n = 15). Cattle were housed in two pens, had ad libitum access to feed and water, and individual intakes and feeding behavior were monitored using the Insentec feeding system. Cattle were fed treatment diets for 111 d, and every 14 d BW were measured and blood samples were collected. Blood plasma was analyzed for glucose, urea nitrogen, and individual amino acids, and results were analyzed using repeated-measures analysis in SAS. Final BW, average daily gain, gain:feed, and hot carcass weight decreased (P ≤ 0.05) by 2.3%, 7.7%, 7.7%, and 2.6%, respectively, in heifers fed the HEMP diet than in heifers fed the DDGS diet. Net energy for maintenance and gain (Mcal/kg of feed, DM basis), estimated based on heifer intake and performance, were greater (P = 0.02) for the DDGS diet than for the HEMP diet. All other performance and carcass characteristics were not different (P ≥ 0.20) between treatments. Heifers fed the HEMP diet had greater (P < 0.05) plasma urea nitrogen concentration in samples from each collection day compared with heifers fed the DDGS diet, although there was a treatment-by-day interaction (P < 0.01) because of variability in the magnitude of treatment differences over time. Plasma glucose concentration was not influenced (P = 0.17) by dietary treatment. Plasma concentrations of total amino acids, nonessential amino acids, and essential amino acids were not different between treatments (P ≥ 0.09), although there were several interactions between treatment and day (P ≤ 0.04) for individual amino acids. These data suggest that hempseed cake has a lower net energy for maintenance and gain relative to DDGS when adequate metabolizable protein is supplied, while still providing adequate nutrition to support the acceptable performance of finishing cattle.

This experiment evaluated the effects of dietary inclusion of hempseed cake in comparison with dried corn distillers grains plus solubles (DDGS) on growth performance, feeding behavior, carcass characteristics, and blood parameters in finishing heifers. Angus-crossbred heifers were assigned randomly to one of two treatments (20% hempseed cake [HEMP] or 20% DDGS; dry matter basis) and were fed for 111 d until slaughter. Heifers receiving the DDGS treatment had greater final body weights, average daily gain, gain efficiency, dietary concentration of net energy for maintenance and gain, and carcass weight than heifers fed the HEMP treatment. All other carcass characteristics, as well as feeding behavior, were not influenced by treatment. Plasma urea nitrogen was greater in heifers fed the HEMP diet compared with the DDGS diet, while glucose was not influenced by treatment. Several plasma amino acid concentrations were influenced by treatment. Although the inclusion of hempseed cake decreased growth performance, it could be a viable alternative feed source for cattle.

Evaluation of the effects of biochar on diet digestibility and methane production from growing and finishing steers.

The objectives of these studies were to evaluate the effects of biochar (0%, 0.8%, or 3% of diet dry matter) on diet digestibility and methane and carbon dioxide production from cattle on growing and finishing diets. The growing diet consisted of 21% brome hay, 20% wheat straw, 30% corn silage, 22% wet distillers grains plus solubles, and 7% supplement. The finishing diet consisted of 53% dry-rolled corn, 15% corn silage, 25% wet distillers grains plus solubles, and 7% supplement. In both trials biochar replaced fine ground corn in the supplement. Six crossbred steers (initial body weight [BW] 529 kg; SD = 16 kg) were used in both the growing and finishing trial. The growing diets were evaluated over 6 periods followed by the finishing trial with 3 periods. Digestibility measures were taken over 4 d after at least 8 d of adaptation to diets followed by 2 d of gas emission measurements using headbox calorimeters. Dry matter intake (DMI) was not affected (P ≥ 0.43; 7.91 kg/d) by biochar inclusion in the growing study and increased quadratically (P = 0.07) in the finishing study with 0.8% biochar inclusion having the greatest DMI (12.9 kg/d). Organic matter (OM) and neutral detergent fiber (NDF) digestibility increased quadratically (P = 0.10) in the growing study whereas OM digestibility tended to linearly decrease (P = 0.13) and NDF digestibility was not affected (P ≥ 0.39) by biochar inclusion in the finishing diet. Digestible energy intake (Mcal/d) was not affected (P ≥ 0.25) by biochar inclusion in the growing or finishing study. Methane production (g/d) tended to decrease quadratically (P = 0.14) in the growing study and was decreased 10.7% for the 0.8% biochar treatment relative to the control. There were no statistical differences in methane production (g/d) in the finishing study (P ≥ 0.32) but cattle on the 0.8% biochar treatment produced numerically less (9.6%) methane than the control. Methane production as g/kg DMI of the 0.8% biochar treatment relative to the control was numerically reduced 9.5% and 18.4% in the growing and finishing studies, respectively (P ≥ 0.13). Carbon dioxide production (g/d and g/kg of intake) quadratically decreased (P ≤ 0.06) in the growing study but was not affected by treatment in the finishing study (P ≥ 0.34). Although biochar is not a U.S. Food and Drug Administration -approved feed for cattle, the initial research shows potential as a methane mitigation strategy in both growing and finishing diets.