Effects of post-weaning supplementation of immunomodulatory feed ingredient on body weight and cortisol concentrations in program-fed beef heifers

Journal Pre-proof Effects of post-weaning supplementation of immunomodulatory feed ingredient on body weight and cortisol concentrations in programmed fed beef heifers J. Danielo, K.J. McCarty, J.E. Tipton, R.E. Ricks, N.M. Long PII:

S0739-7240(19)30106-7

DOI:

https://doi.org/10.1016/j.domaniend.2019.106427

Reference:

DAE 106427

To appear in:

Domestic Animal Endocrinology

Received Date: 28 May 2019 Revised Date:

26 November 2019

Accepted Date: 19 December 2019

Please cite this article as: Danielo J, McCarty KJ, Tipton JE, Ricks RE, Long NM, Effects of postweaning supplementation of immunomodulatory feed ingredient on body weight and cortisol concentrations in programmed fed beef heifers, Domestic Animal Endocrinology (2020), doi: https:// doi.org/10.1016/j.domaniend.2019.106427. This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. © 2019 Published by Elsevier Inc.

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Effects of post-weaning supplementation of immunomodulatory feed ingredient on body weight

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and cortisol concentrations in programmed fed beef heifers*

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J. Danielo,* K. J. McCarty*, J. E. Tipton*, R. E. Ricks*, N. M. Long*,1

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*Department of Animal and Veterinary Sciences, Clemson University, Clemson, SC 29634

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Corresponding author: [email protected]

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ABSTRACT: The objective of this study was to determine the effects of an immunomodulatory

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feed ingredient during post-weaning on growth and cortisol concentrations of beef heifers.

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Commercial Angus heifers (n = 72) from two AI sires were blocked (n = 9) by BW and

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randomly assigned to one of two pens per block. Each pen (4 heifers/pen) per block was

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randomly assigned to one two treatments. Heifers were fed a commercial total mixed ration

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(TMR) twice daily from d 0 to 60 to gain 0.75 kg/day. The feed was top-dressed once daily with

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either 72g of Celmanax (CEL) or corn germ (CON) per pen. Body weight (BW) was collected

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on d -1, 0, 15, 30, 45, 60 and 61. Blood samples were collected on d 0, 15, 30, 45 and 60. Two

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heifers per pen (n = 32) were randomly selected for a transportation challenge to evaluate stress

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response on d 62 or 63 of the study. Sixteen heifers (n = 8 CEL; n = 8 CON) were randomly

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selected for a corticotropin releasing hormone/arginine vasopressin (CRH/AVP) challenge and

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an intravenous glucose tolerance testing (IVGTT) on d 64 and 67 of the study. Pen was the

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experimental unit and data was analyzed by ANOVA or repeated measures analysis as

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appropriate. Feed efficiency and BW gain were increased (P = 0.04) in CEL heifers compared to

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CON heifers. Serum cortisol concentrations were decreased (P < 0.01) in CEL heifers compared

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to CON heifers on d 30 to 60 post-weaning. Serum cortisol concentrations were decreased (P <

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0.05) in CEL heifers compared to CON heifers throughout the transportation challenge. Serum

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cortisol concentrations were decreased (P < 0.05) in CEL heifers compared to CON heifers

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during the CRH/AVP challenge from 60 to 150 min post infusion. Treatment did not influence

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(P = 0.29) plasma insulin or glucose (P = 0.63) concentrations during the IVGTT. In summary,

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supplementation of Celmanax post-weaning increased BW gain and reduced cortisol

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concentrations in challenged beef heifers.

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Key words: immunomodulatory, cortisol, beef heifers, post weaning, Celmanax, yeast

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supplement

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1. Introduction Weaning is one of the most stressful periods of life for beef calves and is associated with

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impaired performance, such as decreased BW and average daily gain (ADG) [1,2], as well as

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increased concentrations of cortisol and norepinephrine [3,4]. During this time, calves are

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removed from the dams and, in most cases, are placed in a foreign or new environments made up

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of unfamiliar surroundings. For example, calves are exposed to stressors such as: dietary

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changes, food deprivation, transportation to new facilities, exposure to diseases and competition

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with other animals. Previously in 2015, cattle operations had a total death loss of 2,144,000

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calves due to respiratory problems (26.9%), calving related problems (17.8%) and digestive

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problems (15.4%) [5]. The total value of beef calf losses due to non-predator causes was over

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$1.5 billion [5], as calves are more susceptible to diseases when introduced to chronic stressors

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[4].

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As a result of environmental stressors during weaning, calves experience negative effects

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on immune function [6], digestive function [7], and altered hormone concentrations [8].

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Immunity may be modulated by manipulation of the hypothalamic-pituitary-adrenal (HPA) axis.

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During a period of stress or trauma, the nervous system initiates a physiological response in

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which the hypothalamus releases corticotropic releasing hormone (CRH) that stimulates the

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anterior pituitary to release adrenocorticotropic hormone (ACTH) into the bloodstream to

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ultimately release cortisol from the adrenal cortex. In previous studies, increased cortisol

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concentrations were negatively correlated with milk and carcass characteristics [9,10] as well as

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overall suppressed performance. Therefore, an immunomodulatory feed ingredient that alters the

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animal’s response to stress and improves overall function could help combat known profit and

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performance loss during the weaning period. The immunomodulatory feed ingredient Celmanax

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(Arm & Hammer Animal Nutrition, Church & Dwight Company, Princeton, NJ) is a killed yeast

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cell wall derivative that consists of biologically active refined functional carbohydrates

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including mannose, mannan-oligosaccharides and beta glucans. Previously, Celmanax has been

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observed to increase weight gain in beef calves [11] and finishing phase beef steers [12], and

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improve milk quality and mammary health of dairy cattle [13]. Interestingly, supplementation of

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yeast products have been observed to alter blood glucose in cattle. Contrasting results have been

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observed between studies, such as increased blood glucose in postpartum dairy cows [14], but

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decreased in transition dairy cows [15]. Supplementation of an immunomodulatory product

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derived from yeast, may also result in physiological differences between animals. The objective

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of this study was to evaluate the effects of supplementation of an enzymatically hydrolyzed yeast

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product on growth and performance, feed efficiency and cortisol concentrations of programmed

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fed beef heifers.

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2. Materials and Methods

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2. 1 Animal Care and Treatments

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Animal care and use were according to protocols approved by the Clemson University Institutional Animal Care and Use Committee (AUP #2017-048).

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Heifers (n = 72) born from primiparous and multiparous cows bred to two AI sires, were

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reared in two pasture groups. Approximately 60 d prior to weaning, all heifers were vaccinated

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with Ultrabac® 7 (Zoetis Inc., Kalamazoo, MI) and Bovi-Shield Gold FP® 5 (Zoetis Inc.,

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Kalamazoo, MI). They received boosters of the same vaccinations on the day of weaning.

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Heifers were weaned at the same location (Clemson University Beef Cattle Farm, Pendleton, SC)

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via traditional weaning strategies at 227 ± 7 days of age (d 0) on September 2, 2017 with an

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average temperature of 25°C with 67% humidity. Heifers were then stratified by BW and sorted

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into nine weight blocks with a replicate on each side of the barn. Heifers were sorted into groups

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of four and then penned (4.8 m x 13.7 m; 18 pens total at 4 heifers per pen). Pen within weight

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block were randomly assigned to one of two treatments 1) Celmanax (Cel) or 2) corn germ

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(Con). Over an adaption period of two weeks, hay was decreased and heifers were transitioned

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from a ration composed of 50% hay (11.8% CP, 1.01 NEm (Mcal/kg) and 0.31 NEg (Mcal/kg))

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and 50% total mixed ration (TMR) (d 0) to a complete TMR ration with no hay (25% corn gluten

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feed pellets, 22.35% soyhull pellets, 51% peanut hull pellets, 1.5% calcium 0.15% trace mineral

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and Vitamin A , D and E premix and 28 g of Rumenson/ton. Analysis 13.8 % CP, 1.15 NEm

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(Mcal/kg) and 0.53 NEg (Mcal/kg)) with ad libitum access to water via automatic waterers.

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Amount of TMR provided per pen were calculated based on average BW of weight block.

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Rations were formulated for heifers to gain 0.75 kg a day when fed at 3.6% of BW on an as fed

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basis [16]. Heifers were fed one half of their daily allotment of TMR at 0630 h and 1800 h each

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day from d 0 until d 60 and were top-dressed with either 72g of Celmanax or corn germ per pen

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at 0630 h. Orts were collected every evening before the evening feeding. After the end of the

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feeding trial, heifers continued to receive TMR with treatments up until d 69. Amount of TMR

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provided per weight block were adjusted every 15 d based on BW. No heifer refused to eat

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within 3 min of feed delivery or showed any outward signs of infections during the study.

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Initial BW was taken consecutively over the first two days (d -1 and 0) and averaged to

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serve as initial BW prior to treatments. Body weight and blood samples were collected at 0600 h

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on d 0 prior to feeding and repeated every 15 d. Final BW was taken consecutively over the last

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two days of the feeding trial (d 60 and 61) then averaged. Serum samples were collected in 9 mL

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serum vacutainers (BD Life Sciences, Franklin Lakes, NJ). The tubes were then incubated at

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room temperature for 1 h and then overnight at 4°C. After overnight incubation, serum samples

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were centrifuged at 1800 x g at 4°C for 20 min. Serum was decanted and stored long term in 1.5

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mL eppendorf tubes (Thermo Fisher Scientific, Waltham, MA) at -20°C until analysis. Plasma

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samples were collected in 9 mL EDTA vacutainers (BD Life Sciences, Franklin Lakes, NJ) and

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immediately placed on ice. Within 1.5 h of blood collection, plasma samples were centrifuged at

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1800 x g for 20 min at 4°C. Plasma was stored in 1.5 mL eppendorf tubes (Thermo Fisher

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Scientific, Waltham, MA) at -20°C until analysis.

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2.2 Transportation Challenge

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Two heifers per pen were randomly selected for the transportation challenge. Half of the

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heifers (n = 18; 9 per treatment; 1 heifer per pen) underwent the transportation challenge on d 62

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and the other half underwent the transportation challenge on d 63. Both groups followed the

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same handling and sampling procedures. Beginning at 0600 h (h 0), a serum sample were

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collected and heifers were loaded into a trailer. Heifers were hauled in a 7.3 m x 2.3 m

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gooseneck trailer for 386.2 km (approximately 4 h) on predominantly interstate road conditions.

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Temperature ranged from an average high of 18.3°C to an average low of 7.2°C with no

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precipitation. After heifers were hauled (h 4), they were unloaded and allowed to comingle in the

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working facility, a serum sample was collected, and then heifers were reloaded randomly (6

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heifers at a time) to be hauled an additional 4 h. At h 8, heifers were unloaded, a serum sample

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was collected, and heifers were then placed on a dry lot and allowed to comingle with ad libitum

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access to only water (no food). After 4 h on dry lot (h 12), another serum sample was collected.

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At 0600 h the following morning (h 24), final serum samples were collected. Serum samples

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were collected in Z/9ml serum collection tubes (Sarstedt, Newton, NC) and all serum samples

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were processed using procedures previously described.

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2.3 Intravenous Glucose Tolerance Test (IVGTT)

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Heifers (n = 16) that were not utilized for the transportation challenge (8 per treatment)

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were randomly selected for intravenous glucose tolerance testing (IVGTT) using methods

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previously described [17]. Half of the heifers (n = 8) underwent IVGTT on d 64 and the other

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half underwent IVGTT on d 67. Both groups followed the same handling and sampling

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procedures. Briefly, heifers were removed from feed 18 h prior to IVGTT, with ad libitum access

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to water. A jugular venous catheter (Abbocath, 16 gauges, Abbott Laboratories, North Chicago,

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IL) and extension set were placed aseptically at 0600 h the morning of the IVGTT. Heifers were

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moved to an individual stanchion and allowed to stand for 2.5 h prior to the start of IVGTT. A 10

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ml waste sample was collected before all blood samples were collected. Blood samples (~6 ml)

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were collected at -15 and 0 min before heifers were given a bolus intravenous infusion of a

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sterile 50% dextrose solution (Nova-Tech, Grand Island, NE) at a dose of 0.3 g of dextrose/kg of

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BW. After bolus administration, blood samples were collected at 5, 10, 15, 20, 25, 30, 45, 60, 90

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and 120 min post-infusion. All blood samples were collected into 9 ml blood collection syringes

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(Sarstedt, Newton, NC) containing sodium heparin and placed on ice immediately after

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collection. Within 30 min of collection, samples were processed as previously described.

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2.4 Corticotropin Releasing Hormone/Arginine Vasopressin Challenge (CRH/AVP)

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The CRH/AVP challenge was performed according to methodologies previously stated in

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Carroll et al. [17]. Briefly, immediately following IVGTT, catheters were flushed with saline in

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order to start the CRH/AVP challenge. Approximately 8 mL of blood was collected in serum

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collection tubes at -60, -30 and 0 min before heifers were given a bolus intravenous infusion of

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Bovine corticotropin-releasing hormone and arginine vasopressin (bCRH and AVP) at a dose of

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0.3 µg and 0.1 µg per kg of BW, respectively [17]. Blood samples were collected in Z/9 mL

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serum collection tubes (Sarstedt, Newton, NC) at 30, 60, 90, 120, 150, 180, 210, 240, 270, 300

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and 330 min post-infusion. Serum samples were processed as previously described.

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2.5 Hormone and Metabolite Analysis

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Plasma glucose was measured colorimetrically in triplicate (Liquid Glucose Hexokinase

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Reagent, Pointe Scientific Inc., Canton, MI) using previously published procedures (Long &

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Schafer, 2013) with an intra-assay and inter-assay CV of 2.3 % and 2.8 %, respectively. Plasma

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insulin was measured in duplicate in two assays by Coat-A-Count Insulin RIA (MP Biomedical,

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Santa Ana, CA) using previously validated procedures [18] with an intra-assay and inter-assay

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CV of 8% and 10%, respectively, and a sensitivity of 0.20 ng/mL. Serum cortisol was measured

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in duplicate in using Coat-A-Count cortisol RIA with a sensitivity of 0.5 mg/dL (MP

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Biomedical, Santa Ana, CA) using previously published procedures [18] with an intra-assay and

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inter-assay CV of 7 % and 8 %, respectively.

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2.6 Statistical Analysis

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Sire was initially placed in the model of preliminary statistical analysis that used heifer as

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the experimental unit as a random effect, was found to be non-significant (P < 0.36), and therefor

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had no effect on the experiment and was removed from all further analysis. Pen performance

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(body weight, body weight gain, feed intake, and feed conversion) was analyzed using the

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MIXED procedure of SAS (SAS software version 9.4, SAS Institute, Cary, NC). Pen served as

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the experimental unit, body weight block as a random variable, and the model statement included

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treatment. Biweekly measurements (cortisol, insulin, and glucose) and cortisol concentrations

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during the transportation challenge were analyzed using repeated measures of ANOVA (SAS

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software version 9.4, SAS Institute, Cary, NC). Pen served as the experimental unit, body weight

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block as a random variable, and the model statement included treatment, time, and their

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interactions. Glucose and insulin concentrations during the IVGTT challenge and cortisol

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concentrations during the CRH/AVP were analyzed using repeated measures of ANOVA (SAS

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software version 9.4, SAS Institute, Cary, NC). Heifer served as the experimental unit for these

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challenges. The model statements included treatment, time, and their interactions. Day of IV

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challenge was initially placed into statistical analysis as a random effect, found to be non-

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significant, and was therefore removed from the model. Statistical significance was declared at

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P ≤ 0.05 while a tendency was declared at 0.05 < P ≤ 0.10.

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3. Results and Discussion

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3.1 Feeding Trial

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Heifer BW, feed consumption and feed conversion are depicted in Table 1. Initial,

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midtrial, and final BW were similar (P > 0.50) between treatments. Body weight gain during the

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feeding trial was greater (P = 0.04) in CEL heifers compared to CON heifers. Total pen feed

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intake was similar (P = 0.987) between treatments. Feed conversion was decreased (P = 0.036)

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in CEL heifers compared to CON heifers. A previous study observed increased ADG and feed

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efficiency throughout a feedlot receiving period in beef steers supplemented with Celmanax

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during either preconditioning or feedlot receiving phases [12]. Kara et al. [19] supplemented

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Holstein Friesian calves with Celmanax from d 5 to 56 of age and observed increased BW and

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ADG (3.70% and 6.66%, respectively) which was similar to the current study. However, Kara et

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al. [19] also observed increased average daily feed intake (10.97%) which differed from the

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current study in which no difference was observed for pen feed intake. This is probably due to

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the fact that our heifers were programmed fed and not given ab libitum access to feed, if they

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would have been given ab libitum access to feed we may have had intake and greater BW gain

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differences. Contrasting results were exhibited in newly received beef heifers supplemented with

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Celmanax for 35 d in which treated heifers observed increased DMI and tended to have greater

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ADG [11]. Results from the current study may not be restricted to a species specific impact on

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performance. Specifically, in lambs, Celmanax supplementation for 82 d with a 14 d adaptation

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period increased weight gain and roughage intake throughout the study, as well as DMI [20].

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Additionally, similar results have been observed in other enzymatically hydrolyzed yeast

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products. Salinas-Chavira [21] supplemented 168 steers with an enzymatically hydrolyzed yeast

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product over a 336 d period and observed increased ADG, final carcass weight, and DMI.

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Treatment did not affect plasma insulin (P < 0.82) or glucose (P < 0.34) concentrations

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during the 60 d feeding trial (Figure 1). Contrasting results have been observed in newly-

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received feedlot heifers supplemented with a yeast cell wall product derived from

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Saccharomyces cerevisiae for 52 d in which increased insulin concentrations were observed on

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day 37, prior to a lipopolysaccharide (LPS) challenge [22]. Results from the current study differ

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from data previously published by Word [23] in which 32 British crossbred beef heifers received

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a live yeast plus yeast cell wall supplement for 31 d following feedlot receiving. The

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supplemented beef heifers underwent a combined viral-bacterial respiratory disease challenge

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and tended to observe a treatment by time interaction for glucose in which treated heifers

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observed increased serum glucose concentrations from 1 h to 6 h following the challenge

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compared to untreated beef heifers [23]. Interestingly, at 6 h post-challenge, the serum glucose

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concentrations were similar between untreated and treated heifers which corresponds with results

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of the current study [23]. Glucose concentrations did not differ in the current study regardless of

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natural causes of stress from the time of weaning or until the time stress was imposed via

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challenges. Whereas, Yuan et al. [24] observed increased β-hydroxybutyrate and that glucose

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concentrations tended to decrease in 40 Holsteins that received Celmanax supplementation 21 d

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prepartum to 42 d postpartum. Treated cows also observed a tendency for increased percentages

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of milk fat, protein and lactose [24]. Celmanax may play a role in metabolic pathways associated

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with gluconeogenesis and glucose utilization.

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Serum cortisol concentrations throughout the feeding trial are depicted in Figure 2.

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Throughout the 60 d feeding trial, serum cortisol concentrations increased (P = 0.049) over time,

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however, concentrations were decreased (P = 0.002) in CEL heifers compared to CON heifers

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throughout the trial. Similar results were observed in Karan Fries heifers receiving a yeast

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supplement (fat plus yeast, niacin, zinc, and chromium) for 90 d during hot, humid months in

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which blood plasma cortisol concentrations were decreased [25]. Additionally, Zaworski et al.

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[26] observed that Holstein cows supplemented with a Saccharomyces cerevisiae fermentation

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products tended to have decreased cortisol concentrations at d 1 and 3 post-partum. Similar

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characteristics have been observed in post-weaned beef calves as Marques et al. [27] observed

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plasma cortisol concentrations that averaged 20 ng/mL (2 µg/dL) from 283 and 290 days of age.

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Increased cortisol concentrations may be associated with decreased performance in livestock

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irrespective of species or production system type. Previously, increased concentrations of fecal

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glucocorticoid metabolite (11,17-dioxoandrostanes) were observed in dairy cows experiencing

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acute heat stress [28], which is associated with decreased milk production [29] and reproductive

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success [30]. This may also be due to the fact that increased milk cortisol concentrations are

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negatively correlated with milk protein percentage and milk solid-not-fat-percentage in Holstein

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cows [9]. Increased cortisol concentrations have also been positively correlated with carcass fat

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content and negatively correlated with carcass lean meat content in pigs [10].

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3.2 Transportation Challenge

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Serum cortisol concentrations in response to the transportation challenge are depicted in

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Figure 3. Serum cortisol concentrations were decreased (P < 0.05) in CEL heifers compared to

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CON heifers throughout the transportation challenge. In recent experiments, cattle observed

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increased serum cortisol concentrations following transportation [31]. Beef steers supplemented

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with Celmanax and subjected to a transportation challenge on d 30 observed transient increased

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cortisol concentrations [12]. The stress associated with transportation may elicit a negative

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immune response in cattle. Previously, Celmanax has been observed to increase weight gain in

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beef calves [11] and finishing phase beef steers [12], and improve milk quality and mammary

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health of dairy cattle [13]. In all of the previous experiments evaluating the use of an

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enzymatically hydrolyzed yeast product, authors speculated involvement of the immune system.

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In the current study, the concomitant response by the immune system and HPA axis release of

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cortisol induced by environmental conditions, such as transportation, supports the idea that

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cortisol concentrations are associated with immune function. Future investigation of components

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of the immune system, such as cytokine activity, and how they interact with physiological

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functions (i.e. digestive) under stressful conditions in cattle are warranted.

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3.3 IVGTT Challenge

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Plasma glucose and insulin concentrations during the IVGTT are depicted in Figure 4 (A

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and B, respectively). Treatment did not influence plasma insulin (P = 0.29) or glucose (P = 0.63)

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concentrations during the IVGTT challenge. A main effect of time was observed in which

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plasma glucose and insulin concentrations decreased (P < 0.0001) throughout the IVGTT

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challenge. Similar results were seen when 24 growing beef steers were supplemented with a

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Chromium yeast product and subjected to an IVGTT, with no differences in basal glucose

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observed [32]. Conversely, Sanchez et al. [33] observed decreased glucose concentrations post

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infusion in Omnigen-AF® supplemented beef heifers that underwent an IVGTT challenge.

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However, similar to the current study, insulin concentrations were not influenced by treatment

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[33]. In a study performed by Kneeskern [34] chromium propionate supplementation to Angus-

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cross steers tended to increase baseline glucose concentrations, while no differences were

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observed for insulin. Contrasting results have been observed in other cattle studies, such as

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increased blood glucose in postpartum dairy cows [14] and decreased blood glucose in transition

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dairy cows [15]. Contrasting results were also observed in Landrace x Yorkshire cross pigs

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supplemented with a high Chromium yeast supplement for 23 to 30 d [35]. Results from an

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IVGTT challenge concluded that treated pigs observed decreased plasma glucose concentrations

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(5 to 20 min post infusion) and increased insulin concentrations (0 and 2 min prior to the

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completion of dextrose infusion [35]. Variation of results between the mentioned experiments

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may be due to the species difference, feeding duration, or the addition of Chromium in the yeast

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supplement. The inclusion of an IVGTT challenge in the current study was not intended to

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induce stress, so much as observe physiological differences between animals, in which none

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were observed in the current study as a result of supplementation of a yeast product.

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3.4 CRH/AVP Challenge

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Serum cortisol concentrations in response to the corticotrophin-releasing

281

hormone/arginine vasopressin challenge are depicted in Figure 5. A treatment by time interaction

282

(P < 0.001) was observed for serum cortisol during the CRH/AVP challenge. Serum cortisol

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concentrations were decreased (P < 0.05) from 60 to 150 min post infusion and increased (P <

284

0.001) at 210 min post infusion in CEL heifers compared to CON heifers. It has been previously

285

demonstrated that cortisol may have immunomodulatory-like effects. For example, during a

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CRH/VP challenge in beef heifers, serum cortisol concentrations and pro-inflammatory

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cytokines were increased due to activation of the hypothalamic pituitary adrenal [17].

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Additionally, holstein cows supplemented with Omnigen-AF®, a yeast containing additive,

289

underwent a CRH, Vasopressin (VP) and ACTH challenge and observed increased neutrophil

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and white blood cell concentrations [36]. Following an LPS challenge, steers supplemented with

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live yeast plus yeast cell wall observed depressed cortisol concentrations compared to control

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steers by 26.6 ng/mL (2.62 µg/dL) [37]. Similar results were observed in the current study even

293

though a killed yeast cell wall product was utilized rather than a live yeast product, while other

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studies have observed differences across varieties of yeast cell wall products. Finck et al. [37],

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supplemented newly weaned crossbred steers with three different yeast cell products (live yeast,

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yeast cell wall, and live yeast plus yeast cell wall), in which steers supplemented with live yeast

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plus yeast cell wall and live yeast observed decreased rectal temperatures and yeast cell wall

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treated animals did not [37]. In the current study, a CRH/AVP challenge was implemented to

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evaluate if the supplementation of an immunomodulatory feed ingredient impacted the HPA

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axis. In recent experiments by Hall et al. [38], lactating dairy cows supplemented with Omnigen-

301

AF® at 56 g per head for 52 d observed reduced cortisol concentrations, but elevated plasma

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ACTH under acute heat stress conditions. While lactating dairy cows supplemented with

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Omnigen-AF® at 56 g per head for 70 d observed reduced cortisol concentrations in both

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thermoneutral and acute heat stress conditions following ACTH and CRH/VP challenges [39].

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Reduced cortisol concentrations in the current study reflect a potential interaction between the

306

hypothalamus and pituitary rather than directly with the adrenal gland based on CRH activity. As

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an adrenocorticotropic hormone (ACTH) challenge was not implemented in the current study to

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determine the direct impact of yeast supplementation on the adrenal glands. However, based off

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of previous observations in dairy cattle and the current study in beef cattle, activation of the HPA

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axis due to stressors may be mitigated by the effects of immunomodulatory supplements,

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particularly at the level of the hypothalamus and pituitary.

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4. Conclusions

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In the current study, Celmanax supplementation increased BW gain, decreased feed

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conversion and serum cortisol concentrations throughout the 60 d feeding trial and decreased

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cortisol concentrations throughout a transportation challenge in program fed beef heifers.

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Activation of the HPA axis due to stressors may be mitigated by the effects of

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immunomodulatory supplements, thus, future investigations on the mechanism and inter-

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relationship between cortisol and immune function are warranted.

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References

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[1] Arthington JD, Spears JW, Miller DC. The effect of early weaning on feedlot performance

322

and measures of stress in beef calves. Journal of Animal Science. 2005;83:933-939.

323

[2] Preedy GW, Jaeger JR, Waggoner JW, Olson, KC. Effects of early or conventional weaning

324

on beef cow and calf performance in pasture and drylot environments. Journal of Animal

325

Science. 2016;94:610.

326

[3] Lay DC Jr, Friend TH, Randel RD, Bowers CL, Grissom KK, Neuendorff DA, Jenkins

327

OC. Effects of restricted nursing on physiological and behavioral reactions of Brahman calves to

328

subsequent restraint and weaning. Applied Animal Behaviour Science. 1998;5:109-119.

329

[4] Hickey MC, Drennan MM, Earley B. The effect of abrupt weaning of suckler calves on the

330

plasma concentrations of cortisol, catecholamines, leukocytes, acute-phase proteins and in vitro

331

interferon-gamma production. Journal of Animal Science. 2003; 81(11):2847-2855.

332

[5] USDA. Cattle and Calves Death Loss in the United States Due to Predator and

333

Nonpredator Causes. 2015. USDA–APHIS–VS–CEAH. Fort Collins, CO #745.1217

334

[6] Lynch EM, Earley B, McGee M, Doyle S. Effect of abrupt weaning at housing on leukocyte

335

distribution, functional activity of neutrophils, and acute phase protein response of beef calves.

336

BMC Veterinary Research. 2010;6:39.

337

[7] Baldwin RL, McLeod KR, Klotz JL, Heitmann RN. Rumen Development, Intestinal Growth

338

and Hepatic Metabolism In The Pre-and Postweaning Ruminant. Journal of Dairy Science.

339

2003;87:E55-E65.

340

[8] Kim M, Yang J, Upadhaya SD, Lee H, Yun C, Ha JK. The stress of weaning influences

341

serum levels of acute-phase proteins, iron-binding proteins, inflammatory cytokines, cortisol, and

342

leukocyte subsets in Holstein calves. Journal of Veterinary Science. 2011;12:151.

343

[9] Fukasawa M, Tsukada H, Kosako T, Yamada A. Effect of lactation stage, season and parity

344

on milk cortisol concentration in Holstein cows. Livestock Science. 2008;113:280–284.

345

[10] Foury A, Devillers N, Sanchez M-P, Griffon H, Roy PL, Mormède P. Stress hormones,

346

carcass composition and meat quality in Large White × Duroc pigs. Meat Science.

347

2005;69(4):703-707.

348

[11] Ponce CH, Schutz JS, Elrod CC, Anele UY, Galyean ML. Effects of dietary

349

supplementation of a yeast product on performance and morbidity of newly received beef

350

heifers. Professional Animal Scientist. 2012; 28(6):618-622.

351

[12] Silva LGT, Cooke RF, Schubach KM, Brandão AP, Marques RS, Schumaher TF, Moriel P,

352

Bohnert DW. Supplementing a yeast-derived product to enhance productive and health responses

353

of beef steers. Animal. 2018;12(8):1576-1583.

354

[13] Nocek JE, Holt MG, Oppy J. Effects of supplementation with yeast culture and

355

enzymatically hydrolyzed yeast on performance of early lactation dairy cattle. Journal of dairy

356

science. 2011; 94(8): 4046-4056.

357

[14] Nocek JE, Kautz WP. Direct-fed microbial supplementation on ruminal digestion, health,

358

and performance of pre-and postpartum dairy cattle. Journal of Dairy Science. 2006; 89(1): 260-

359

266.

360

[15] Yuan K, Liang T, Muckey MB, Mendonça LGD, Hulbert LE, Elrod CC, Bradford BJ. Yeast

361

product supplementation modulated feeding behavior and metabolism in transition dairy cows.

362

Journal of Dairy Science. 2015a; 98(1): 532-540.

363

[16] NRC. 2000. Nutrient requirements of beef cattle. 7th rev. ed. Natl. Acad. Press,

364

Washington, DC.

365

[17] Carroll JA, McArthur NH, Welsh Jr TH. In Vitro and In Vivo Temporal Aspects of ACTH

366

Secretion: Stimulatory Actions of Corticotropin‐Releasing Hormone and Vasopressin in Cattle.

367

Journal of Veterinary Medicine Series A. 2007; 54(1): 7-14.

368

[18] Long NM, Schafer DW. Sex effects on plasma leptin concentrations in newborn and

369

postnatal beef calves. Prof Anim Sci. 2013; 29:601–5.

370

[19] Kara C, Cihan H, Temizel M, Catik S, Meral Y, Orman A, Yibar A, Gencoglu H. Effects of

371

Supplemental Mannanoligosaccharides on Growth Performance, Faecal Characteristics and

372

Health in Dairy Calves. Asian-Australasian Journal of Animal Science. 2015; 28:1599-1605.

373

[20] Nde FF, Verla NS, Michael C, Ahmed MA Effect of Celmanax on feed intake, live weight

374

gain and nematode control in growing sheep. African Journal of Agricultural Research. 2014;

375

9:695-700.

376

[21] Salinas-Chavira J, Arzola C, González-Vizcarra V, Manríquez-Núñez OM, Montaño-

377

Gómez MF, Navarrete-Reyes JD, Raymundo C, Zinn RA. Influence of Feeding Enzymatically

378

Hydrolyzed Yeast Cell Wall on Growth Performance and Digestive Function of Feedlot Cattle

379

during Periods of Elevated Ambient Temperature. Asian-Australasian Journal of Animal

380

Science. 2015; 28:1288-1295.

381

[22] Burdick Sanchez NC, Young TR, Carroll JA, Corley JR, Rathmann RJ, Johnson BJ. Yeast

382

cell wall supplementation alters aspects of the physiological and acute phase responses of

383

crossbred heifers to an endotoxin challenge. Innate Immunity. 2013; 19(4):411-419.

384

[23] Word AB, Broadway PR, Burdick Sanchez NC, Roberts SL, Richeson JT, Liang YL,

385

Holland BP, Cravey MD, Corley JR, Ballou MA, Carroll JA. Immune and metabolic responses

386

of beef heifers supplemented with Saccharomyces cerevisiae to a combined viral-bacterial

387

respiratory disease challenge. Translational Animal Science. 2019; 3(1):135-148.

388

[24] Yuan K, Mendonça LGD, Hulbert LE, Mamedova LK, Muckey MB, Shen Y, Elrod CC,

389

Bradford BJ. Yeast product supplementation modulated humoral and mucosal immunity and

390

uterine inflammatory signals in transition dairy cows. Journal of Dairy Science. 2015b; 98:3236-

391

3246.

392

[25] Purwar V, Oberoi PS, Dang AK. Effect of feed supplement and additives on stress

393

mitigation in Karan Fries heifers. Veterinary World. 2017; 10(12):1407-1412

394

[26] Zaworski EM, Shriver-Munsch CM, Fadden NA, Sanchez WK, Yoon I, Bobe G. Effects of

395

feeding various dosages of Saccharomyces cerevisiae fermentation product in transition dairy

396

cows. Journal of Dairy Science. 2014; 97:3081-3098.

397

[27] Marques RS, Cooke RF, Rodrigues MC, Cappellozza BI, Mills RR, Larson CK, Moriel P,

398

Bohnert DW. Effects of organic or inorganic cobalt, copper, manganese, and zinc

399

supplementation to late-gestating beef cows on productive and physiological responses of the

400

offspring. Journal of animal science. 2016; 94(3):1215-1226.

401

[28] Rees A, Fischer‐Tenhagen C, Heuwieser W. Effect of Heat Stress on Concentrations of

402

Faecal Cortisol Metabolites in Dairy Cows. Reproduction in Domestic Animals. 2016;51:392-

403

399.

404

[29] Collier RJ, Eley RM, Sharma AK, Pereira RM, Buffington DE. Shade Management in

405

Subtropical Environment for Milk Yield and Composition in Holstein and Jersey Cows. Journal

406

of Dairy Science. 1981; 64:844-849.

407

[30] Ingraham RH, Gillette DD, Wagner WD. Relationship of Temperature and Humidity to

408

Conception Rate of Holstein Cows in Subtropical Climate. Journal of Dairy Science. 1974;

409

57:476-481.

410

[31] Chulayo AY, Bradley G, Muchenje V. Effects of transport distance, lairage time and

411

stunning efficiency on cortisol, glucose, HSPA1A and how they relate with meat quality in

412

cattle. Meat Science. 2016; 117:89-96.

413

[32] Swanson KC, Harmon DL, Jacques KA, Larson BT, Richards CJ, Bohnert DW, Paton SJ.

414

Efficacy of chromium-yeast supplementation for growing beef steers. Animal Feed Science and

415

Technology. 2000; 86:95-105.

416

[33] Sanchez NB, Carroll JA, Broadway PR, Schell TH, Puntenney SB, McLean DJ.

417

Supplementation of Omnigen-AF alters the metabolic response to a glucose tolerance test in beef

418

heifers. Journal of Animal Science. 2017; 95:215.

419

[34] Kneeskern SG, Dilger AC, Loerch SC, Shike DW, Felix TL. Effects of chromium

420

supplementation to feedlot steers on growth performance, insulin sensitivity, and carcass

421

characteristics. Journal of Animal Science. 2016;94:217-226.

422

[35] Guan X, Matte JJ, Ku PK, Snow JL, Burton JL, Trottier NL. High Chromium Yeast

423

Supplementation Improves Glucose Tolerance in Pigs by Decreasing Hepatic Extraction of

424

Insulin. The Journal of Nutrition. 2000; 130(5):1274-1279.

425

[36] Sanchez NC, Carroll JA, Mcbride ML, Ortiz XA, Collier JL, Chapman JD, Mclean D,

426

Collier RJ. 2015. OmniGen-AF alters rectal temperature (RT) and leukocyte profiles in dairy

427

cows exposed to heat stress (HS) following acute activation of the stress axis. Journal of Animal

428

Science Supplement. 2015; 93.

429

[36] Finck DN, Ribeiro FRB, Burdick NC, Parr SL, Carroll JA, Young TR, Bernhard BC, Corley

430

JR, Estefan AG, Rathmann RJ, Johnson BJ. Yeast supplementation alters the performance and

431

health status of receiving cattle. The Professional Animal Scientist. 2014; 30(3):333-341.

432

[37] Hall LW, Villar F, Chapman JD, McLean DJ, Long NM, Xiao Y, Collier JL, Collier RJ. An

433

evaluation of an immunomodulatory feed ingredient in heat-stressed lactating Holstein cows:

434

Effects on hormonal, physiological, and production responses. Journal of dairy science. 2018;

435

101(8): 7095-7105.

436

[38] McBride ML, Sanchez NB, Carroll JA, Broadway PR, Ortiz XO, Collier JL, McLean D,

437

Chapman JD, Kattesh HG, Collier RJ. 1128 OmniGen-AF® reduces basal plasma cortisol as

438

well as cortisol release to adrencocorticotropic hormone or corticotrophin releasing hormone and

439

vasopressin in lactating dairy cows under thermoneutral or acute heat stress conditions. Journal

440

of Animal Science. 2016; 94(suppl 5):541-542.

441

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Table 1. Heifer BW, feed consumption, and feed conversion efficiency of heifers fed a total mixed ration top dressed with corn germ meal (CON) or Celmanax (CEL) throughout the 60 day feeding trial Item CON CEL SEM P-Value N 9 9 Initial BW, kg 242 243 8 0.983 D 30 of trial BW, kg 260 263 8 0.878 D 60 of trial BW, kg 282 286 5 0.584 BW gain during trial, kg 40 43 1 0.040 Pen feed intake, kg 2273 2272 43 0.987 Feed conversion, kg intake/kg BW 14.65 13.35 0.39 0.036 gain 1 BW at d 30 of the trial 2 BW at d 60 of the trial 3 BW gain by the end of the 60 d trial Data presented LSM ± SEM.

451

Figure Legends

452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470

Figure 1. Plasma glucose concentrations (mg/dL; a) and insulin concentrations (ng/mL; b) of heifers fed corn germ meal (CON) or Celmanax (CEL) throughout the 60 day feeding trial.

471

Figure 2. Serum cortisol concentrations (µg/dL) of heifers fed corn germ meal (CON) or Celmanax (CEL) throughout the 60 day feeding trial. An asterisk (*) signifies a significant difference between treatments at that time point P ≤ 0.05. Figure 3. Serum cortisol concentrations (µg/dL) of heifers fed corn germ meal (CON) or Celmanax (CEL) during a transportation challenge following the 60 day feeding trial. An asterisk (*) signifies a significant difference between treatments at that time point P ≤ 0.05. Figure 4. Plasma glucose concentrations (mg/dL; a) and insulin (ng/mL; b) concentrations of heifers fed corn germ meal (CON) or Celmanax (CEL) during the intravenous glucose tolerance test following the 60 day feeding trial. Figure 5. Serum cortisol concentrations (µg/dL) of heifers fed corn germ meal (CON) or Celmanax (CEL) during corticotrophin-releasing hormone/ arginine vasopressin challenge following the 60 day feeding trial. An asterisk (*) signifies a significant difference between treatments at that time point P ≤ 0.05.

472 473

Figure 1.

Plasma insulin, ng/mL

474 3.0

2.5

2.0

1.5

1.0 0

15

30

45

Day

475 476

Trt P = 0.82, time P < 0.001

60

477

Figure 2. 5

Serum cortisol, µ g/dL

*

4

*

*

30

45

3 2 1 0 0

15

Day 478 479

Trt P = 0.002, time P = 0.049

60

480

Figure 3.

Serum cortisol, µ g/dL

4

3

*

*

* *

*

2

1

0 0

4

8

12

16

20

Hours 481 482

Trt P < 0.0001, time P = 0.96

24

483

Figure 4.

Plasma insulin, ng/mL

484 15

10

5

0 -15

0

15

30

45

60

75

90

Min

485 486

Trt P = 0.63, time P < 0.0001

105 120

487 488

Figure 5.

Serum cortisol, µ g/dL

10

*

8

* * *

6

*

4 2 0 -60

0

60

120

180

Hours

489 490

Trt *time P < 0.001

240

300

Highlight for review: • • •

Heifers were programmed fed a commercial TMR twice daily from d 0 to 60 to gain 0.75 kg/day. The feed was top-dressed once daily with either 72g of Celmanax (CEL) or corn germ (CON) per pen. Feeding Celmanax increased heifer BW gain and decreased the amount of feed to get a kg of gain. Feeding Celmanax decreased heifer cortisol concentrations during the feeding period, during a transportation challenge, and during a CRH/AVP challenge.