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Efficacy of oral meloxicam as primary pain mitigation following caustic paste disbudding of three day old Holstein calves

Efficacy of oral meloxicam as primary pain mitigation following caustic paste disbudding of three... Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Efficacy of oral meloxicam as primary pain mitigation following caustic paste disbudding of three day old Holstein calves † ‡ || $ ¶, Kyle J. Karlen, Faith S. Baier, Sara L. Odegard, Ruth M. Baumann, Johann F. Coetzee, || ||,2 Sylvia I. Kehoe, and Kurt D. Vogel † ‡ School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706; Department of Animal Sciences, || Colorado State University, Fort Collins, CO 80523; Department of Animal and Food Science, University of $ ¶ Wisconsin, River Falls, WI 54022; Golden Eagle Lodge, Grand Marais, MN 55604; and College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506 ABSTRACT: The purpose of this study was to in- Plasma was collected and analyzed for cortisol and vestigate the effects of perioperative administration substance P concentrations. Substance P and cor- of oral meloxicam prior to and following the ap- tisol decreased in all animals over time, regardless plication of caustic paste to disbud neonatal dairy of treatment. Mean plasma substance P concentra- calves. Sixty-one 3–4-d-old Holstein heifer calves tion across all time points was greater (P  <  0.05) were randomly assigned to one of four treatment in the SHAM group than M1 or M2 but not dif- groups of 15–16 calves. The treatment groups were: ferent (P > 0.05) than the CONTROL group. The 1) M1, caustic paste disbudding and oral meloxicam MNT and ocular temperatures decreased over time (45 mg) with a placebo 24 h later; 2) M2, treatment across all treatments (P  <  0.05). Mean BW in- M1 followed by a second 45-mg dose of meloxicam creased over time across all treatments (P < 0.05). 24 h later instead of placebo; 3) CONTROL, treat- A significant interaction (P  < 0.05) between treat- ment M1 with placebo in place of meloxicam; and ment and sampling time was observed at 12 h fol- 4) SHAM, sham disbudding with placebo in place lowing treatment application for both mean horn of meloxicam. Infrared thermography was used to bud temperature and the ratio between horn bud quantify eye and horn bud temperatures. Pressure and ocular temperature. Overall, the results of this algometry was used to measure Mechanical noci- study suggest that meloxicam administration at a ceptive threshold (MNT) surrounding the horn dose of 45 mg per animal may have limited influ- bud. Average daily gain and body weight (BW) ence as the primary modulator of pain and inflam- were obtained by weighing each animal throughout matory response in calves that have been disbudded the study and calculating the changes over time. with caustic paste at 3 d of age. Key words: calf, dehorn, disbud, horn, meloxicam, pain © The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. Transl. Anim. Sci. 2019.XX:0-0 doi: 10.1093/tas/txz151 The authors greatly appreciated the hospitality of the com- thoughtful and valuable input regarding the content of our mercial dairy that allowed us to access and use their calves for manuscript. This project was funded by the University of Wis- this study. We would also like to thank E.  Andersen, A.  Bue, consin—River Falls (UWRF) Undergraduate Research, Schol- K. Guden, S. Heck, J. Hollatz, A. Keller, A. Long, C. McClin- arly, and Creative Activity program, UWRF Falcon Scholars tock, K. Schiller, and J. Spoehr for their assistance in sample col- program, and a generous gift from The Kraft Heinz Company. lection and management. We also appreciated the guidance and Corresponding author: kurt.vogel@uwrf.edu assistance of J. Peterson from the Iowa State University PHAST Received June 29, 2019. Lab. In addition, we thank the anonymous reviewers for their Accepted October 1, 2019. 1 Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 2 Karlen et al. INTRODUCTION One potential option for pain mitigation for caustic paste disbudding is meloxicam. Meloxicam One of the most common painful procedures is a long-acting NSAID that is similarly available for North American dairy cattle is removal of both parenterally and orally, has peak availability the horns or horn buds. In 2014, 94.3% of North at approximately 12  h, and has a half-life of 28  h American dairy operations reported routine re- (Coetzee et  al., 2009). This makes meloxicam moval of the horns of heifer calves on their prem- uniquely suited for administration up to 8  h prior ises (National Animal Health Monitoring System to dehorning or shortly before the application of [NAHMS], 2014). Of those dairy operations, 16.4% caustic paste and may make a second dose unneces- utilized caustic paste for disbudding at 2.3  wk of sary until days after the initial dose was adminis- age (NAHMS, 2014). In the same survey, 5.6% of tered as meloxicam may suppress the concentrations the dairy operations that used caustic paste for rou- of prostaglandin E2 (PGE ), an important pros- tine disbudding of dairy calves utilized analgesic taglandin associated with the Cyclooxygenase-2 and/or anesthetic pain mitigation (NAHMS, 2014). (COX-2) inflammatory pathway, for as long as Across all dairy operations that performed routine 3 d (Allen et  al., 2013). To this point in time, no horn removal, pain mitigation use was dependent previous work had been published regarding the on the person that performed the procedure as impact of meloxicam as the primary pain mitiga- 62.7% of veterinarians, 29.2% of employees, and tion therapy for caustic paste disbudding, and the 14.9% of owner/operators reported that they used research on caustic paste disbudding in general is anesthetic and/or analgesic pain mitigation for any limited. As a result, this study was focused on ex- routine horn removal procedures (NAHMS, 2014). ploring the impact of practical meloxicam admin- Caustic paste disbudding has been suggested to istration techniques on dairy calves. The objective cause less acute pain response than cautery disbud- of this study was to investigate the effects of peri- ding (Vickers et al., 2005), and the procedure itself operative administration of oral meloxicam prior has a more innocuous appearance than other horn to and following the application of caustic paste to removal methods (Vickers et  al., 2005). However, disbud neonatal dairy calves. more recent work has suggested that caustic paste may be more noxious than cautery disbudding MATERIALS AND METHODS but deceiving to assess due to the differing meth- ods of application and action (Braz et  al., 2012; Institutional Animal Care and Use Committee Hempstead et al., 2018). Approval Based on behavioral and cortisol responses, peak pain appears to occur within the first 4 h fol- Prior to the commencement of this study, animal lowing caustic paste application, with the poten- use and associated procedures were approved by the tial for chronic pain after that time period (Morisse University of Wisconsin—River Falls Institutional et al., 1995; Stilwell et al., 2009). Winder et al. (2017) Animal Care and Use Committee. The protocol reported that acute pain persisted for a minimum of number associated with this study was A15-16–7. 180 min following the application of caustic paste to disbud 3–5-d-old Holstein calves that were treated Animals and Housing with 0.5 mg/kg subcutaneous meloxicam 15 min be- fore caustic paste application. In the same study, re- Sixty-one female Holstein calves (41.2 ± 0.1 kg gional anesthesia in the form of a lidocaine cornual body weight [BW]) were housed in individual out- nerve block (5 mL per side) effectively reduced acute door hutches on a sand base with a layer of wood pain indication by algometry for approximately shavings and straw bedding inside each hutch (ap- 180  min in calves that were treated with the same proximate area: 2.4 m ) and in an individual out- dose of meloxicam 15 min before caustic paste appli- door exercise space accompanying each hutch cation. Braz et al. (2012) indicated that caustic paste (approximate area: 2 m ). Each calf was fitted disbudding caused severe pain for the first 30 min fol- with a thermal jacket that was worn for the dur- lowing paste application and found tramadol to be ation of the study at the time of separation from ineffective in controlling acute pain associated with her dam. All calves were born and housed on the the procedure. Stilwell et  al. (2008, 2009) reported same commercial dairy farm in Western Wisconsin that flunixin meglumine alone was also ineffective between February and April 2016. Each calf was in controlling a postapplication cortisol increase in separated from her dam within an hour of birth dairy calves following caustic paste application. and fed 3.8 liters of colostrum by gastroesophageal Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Oral meloxicam after caustic paste disbudding 3 tube within 2 h of birth. A second feeding of col- Administration of Oral Meloxicam ostrum was administered by bottle 12–24  h after All calves in this experiment were fed 1.9 li- the initial colostrum feeding. Following the second ters of pasteurized milk after the administration colostrum feeding, each calf was fed 1.9 liters of of meloxicam and collection of measurements at pasteurized whole milk three times daily. All calves each time point. The experimental dose of meloxi- were enrolled in the study and received their as- cam or placebo was administered via balling signed treatments at exactly 3 d of age. The number gun. Three—15  mg meloxicam tablets (Unichem of calves that were enrolled in the study on each Pharmaceuticals (USA) Inc., Rochelle Park, NJ) day varied depending on the number of calves that were loaded into empty gel capsules (size 00, Torpac were born 3 d earlier. The treatments were assigned Inc., Fairfield, NJ) and two capsules at a time were in rolling order such that each treatment was repre- administered to calves in the M1 and M2 treatment sented once in each group of four calves that were groups at the scheduled administration times. Of consecutively enrolled in the study. The specific the two capsules, one capsule was always empty treatment that each calf received was dependent on and served to assist in holding the meloxicam or the chronological birth order and next treatment placebo capsule in the balling gun during passage assignment in the repeating treatment applica- into the esophagus. The second capsule contained a tion order. Farm staff were blind to all treatments. total of 45 mg of meloxicam. Each dose of meloxi- Some project personnel were not blind to the treat- cam provided approximately 1.0 mg meloxicam per ment groups if they were involved in treatment ap- kg of calf BW. Meloxicam or placebo delivery oc- plication. However, the specific outcomes that were curred approximately 5 min before the disbudding assessed were unlikely to be affected by the treat- procedure and just before the feeding 24 h postdis- ment knowledge of those personnel. budding. Calves that received placebo at either or both time points received empty gel capsules via Experimental Design and Treatments balling gun instead of meloxicam in gel capsules. Each calf was assigned to one of four treat- ment groups. Treatment assignments were pro- Disbudding and Sham Disbudding Procedure vided in rolling order as calves were enrolled in the study at 3 d of age. The treatment groups Prior to disbudding treatment application, a were: 1)  M1, caustic paste disbudding and oral battery-operated hair clipper was used to trim the meloxicam (45  mg) with a placebo 24  h later; hair to a length of approximately 1  mm within a 2) M2, treatment M1 followed by a second 45-mg 3-cm radius of the horn buds. A  commercially dose of meloxicam 24 h later instead of placebo; available caustic paste product that contained a 3)  CONTROL, treatment M1 with placebo in mixture of sodium and calcium hydroxide (Dr place of meloxicam; and 4)  SHAM, sham dis- Naylor Dehorning Paste, H.  W. Naylor Company budding with placebo in place of meloxicam. Inc, Morris, NY) or a noncaustic analog (Up and The treatment application and sample collection Up creamy diaper rash ointment, Target Stores, timeline may be observed in Figure 1. At each Minneapolis, MN) was administered to the horn time point, ocular and horn bud thermographs bud. The noncaustic analog was similar in consist- were collected followed by algometry data for the ency to the caustic paste but differed in color. The tissue surrounding the horn buds. On odd-num- caustic paste was light red in color and the non- bered evenings beginning on day 1, BW was also caustic analog was white. Paste, either caustic or recorded in addition to the thermographs and noncaustic, was evenly applied with a gloved hand algometry readings. Thermographs and algom- at a thickness of approximately 1  mm to an area etry readings were collected during the morning on the horn bud that was approximately 18 mm in feedings on days 1–5 and on odd-numbered even- diameter ings for each calf, resulting in measurements for time points −5 min, 0, 12, 24, 48, 60, 72, 96, 108, BW and Average Daily Gain 156, 204, 252, and 300 h from disbudding. For all treatments and measurements except blood col- The BW of each animal involved in this study lection, calves were restrained in sternal recum- was collected from a digital hanging scale (Product bency with one student straddling the calf while #34626, Roughneck Digital Hanging Scale, applying one hand to the base of the head and one Northern Tool + Equipment, Burnsville, MN) to the nose to limit movement of the calf ’s head. equipped with a commercially available sling (Item Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 4 Karlen et al. Caustic Paste applied to M1, M2, and CONTROL. Non-caustic analog applied for SHAM. Meloxicam applied to M2. Meloxicam applied to M1 Placebo and M2. applied to CONTROL, Placebo applied SHAM, and to CONTROL M1. and SHAM. BD BD BD BD BD TH TH TH TH TH TH TH TH TH TH TH TH TH PA PA PA PA PA PA PA PA PA PA PA PA PA BW BW BW BW BW BW BW BW BW BW BW BW BW - 0.5 0 12 24 48 60 72 108 156 204 252 300 Hours Figure 1. This timeline graphic outlines the treatment administration times and sample collection times over the course of the study. The treat- ment groups were: 1) M1: caustic paste disbudding and oral meloxicam (45 mg) with a placebo 24 h later; 2) M2: treatment M1 followed by a second 45-mg dose of meloxicam 24 h later instead of placebo; 3) CONTROL: treatment M1 with placebo in place of meloxicam; and 4) SHAM: sham disbudding with placebo in place of meloxicam. Sample collections are defined as: BD = blood draw, TH = ocular and horn bud infrared therm- ography, PA = pressure algometry, WT = body weight. Time point 0 marks the measurements recorded immediately after treatment application. # 17123, Calf Weighing Sling, Valley Vet Supply, of collection, all whole blood was centrifuged in Marysville, KS) to safely suspend each calf during the original collection tube for cortisol and the so- weight collection. BW was recorded on odd-num- dium EDTA plus benzamidine tubes for substance bered evenings over a 13-d period beginning on the P at 2,500 × g for 15 min. Plasma was transferred day of disbudding and meloxicam treatment ap- to microcentrifuge tubes with disposable transfer plication. To calculate ADG, the total amount of pipettes and then frozen in liquid nitrogen before BW change between BW collection events was div- transferring to storage at −80 °C until analysis via ided by the total number of experimental days that radioimmunoassay at the Iowa State Veterinary elapsed since the previous BW collection event. diagnostic laboratory. Blood Collection Cortisol and Substance P Quantification Whole blood was collected via jugular veni- Plasma substance P and cortisol concentra- puncture with 21-g × 2.54-cm blood collection tions were quantified at the Iowa State University needles (REF 450042, Greiner Bio-One GmbH, Veterinary Diagnostic Laboratory following pro- Kremsmünster, Austria) from all animals in each cedures that were described by Kleinhenz et  al. of the four treatment groups. Blood was collected (2018). A  commercially available kit (catalog # prior to the administration of meloxicam and 06B-256440, Corti-Cote Cortisol RIA Kit, MP caustic paste (time −5 min) to obtain baseline sub- Biomedicals Inc., Santa Ana, CA) was used to es- stance P and cortisol concentrations, in addition to timate plasma cortisol concentrations for each times 24, 48, 72, and 96 h. At each collection time, sample in duplicate following the manufacturer’s a single 10-mL nonadditive blood collection tube procedure. The duplicate values were averaged for (BD, Franklin Lakes, NJ) and a single 6-mL blood each sample prior to statistical analysis. The coef- collection tube with lithium heparin additive (BD, ficient of variation for intra-assay variability was Franklin Lakes, NJ) were collected from each calf. 10.31%. The coefficient of variation for interassay Immediately after collection, 6 mL of whole blood variability was 5.59%. from the nonadditive tube was transferred to a 6-mL Substance P was quantified in nonextracted blood collection tube that contained sodium ethyl- plasma following procedures that were out- enediaminetetraacetic acid (EDTA) additive plus lined by Kleinhenz et  al. (2018), Van Engen et  al. 300  μL of benzamidine solution. Within 30  min (2014), and Liu et  al. (2008). A  two-antibody Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Oral meloxicam after caustic paste disbudding 5 radioimmunoassay (RIA) was used to estimate Kleinhenz et al., 2018). An infrared camera (FLIR plasma substance P concentrations for each sample E8 Thermal Camera, FLIR Systems, Boston, MA) in duplicate. The duplicate values were averaged for was used to collect three thermal images of each each sample prior to statistical analysis. The coef- eye and each horn bud for each animal at the time ficient of variation for intra-assay variability was points identified in Figure 1. Thermographs were 10.55%. The coefficient of variation for interassay collected from a distance of approximately 30.5 cm variability was 27.64%. with the camera focal point in perpendicular align- ment with the target location. A laser tape measure (Bosch GLM 10 Compact Laser Measure, Robert Meloxicam Quantification Bosch GmbH, Stuttgart, Germany) was attached Plasma meloxicam concentrations were quanti- to the thermal camera to facilitate rapid, accurate, fied for calves in the M1 and M2 treatment groups and consistent distance determination. at the Iowa State University Veterinary Diagnostic Ocular thermographs were collected first and Laboratory following procedures similar to those then each calf was blindfolded and allowed to ac- described by Repenning et al. (2013). High-pressure climate to the blindfold while horn bud thermo- liquid chromatography tandem–mass spectrom- graphs were collected prior to pressure algometry. etry (HPLC–MS) was performed with a LTQ ion Stock et  al. (2015) also used a blindfold to assist trap (Thermo Scientific, San Jose, CA) coupled in maintaining stable body and head position dur- with an Agilent 1100 series pump and autosampler ing algometry. Thermographs were analyzed with (Agilent Technologies, Santa Clara, CA) to ana- FLIR Tools software (FLIR Systems, Boston, lyze each sample in single. The instrument injec- MA). For each image, the area of interest (ei- tion volume was set to 20 μL. The mobile phases ther eye or horn bud) was selected via a function were A: 0.1% formic acid in water; and B: 0.1% in FLIR Tools, and the maximum temperature of formic acid in acetonitrile. The gradient curve the selected field was recorded and entered into a began at 25% B with a linear gradient to 95% B in Microsoft Excel spreadsheet and used for statistical 5 min, maintained for 1.75 min, then re-equilibra- analysis. The mean of the three maximum temper- tion to 25% B. The o fl w rate was 0.25 mL/min and atures recorded from the thermal images for each increased to 0.325 mL/min at 5.25 min. Separation eye and each horn bud was ultimately included in was performed with an UltraCore C18 100- × the statistical analyses. 2.1-mm column (Kinetex XB-C18, Phenomenex, Totrrence, CA). Meloxicam eluted at 5.4 min and Pressure Algometry piroxicam eluted at 4.7  min. Four selected reac- tion monitoring transitions each were monitored Pressure algometry has been used to assess the for meloxicam (115, 141, 184, and 352 m/z) and mechanical nociceptive threshold (MNT) of ani- piroxicam (227, 270, 282, and 332 m/z). Sequences mals in previous experiments investigating the pain were batch processed with a method developed in response to disbudding (Heinrich et al., 2010; Allen Xcalibur software (Thermo Scientific, San Jose, et al., 2013; Glynn et al., 2013; Winder et al., 2017; CA). The method automatically identified and in- Adcock and Tucker, 2018). MNT has been used tegrated each peak in each sample and calculated as a measure of sensitivity to physical stimulus; a the calibration curve based on a weighted (1/X) greater MNT value suggests a greater tolerance to quadratic fit. Plasma concentrations of meloxi- force and decreased sensitivity. A  pressure algom- cam in unknown samples were calculated by the eter (Wagner Force Ten FDX 25 Compact Digital Xcalibur software based on the calibration curve. Force Gage, Wagner Instruments, CT) was used The correlation coefficient (r ) was > 0.98 across to apply pressure to three areas anterior to, lateral the concentration range of 5–5,000  ng/mL. QC to, and posterior to each horn bud at the edge of samples were within tolerance of ±15% of the the paste that was applied. A 1-cm flat rubber tip nominal value. accessory was affixed to the algometer. Force was applied at an approximate rate of 1.0  kgf/s until the animal attempted to move its head away from Infrared Thermography the applied pressure, similar to the procedure in Infrared thermography has previously been used Winder et  al. (2017). Maximum force did not ex- to observe the change in temperature of the eye dur- ceed 10 kg force to prevent excessive discomfort for ing painful procedures in several studies (Stewart the calf. The maximum force applied prior to move- et  al., 2008; Allen et  al., 2013; Stock et  al., 2015; ment away from the pressure was recorded for each Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 6 Karlen et al. of the three locations for each of the horn buds and Drug Administration (FDA) as an NSAID at each collection time identified in Figure 1. The for cattle with a label referencing pain treatment. mean MNT for each horn bud at each collection Therefore, any administration of meloxicam falls time was included in the statistical analyses. into the category of extra label drug use (ELDU). A  valid veterinary patient client relationship Statistical Analyses (VCPR) must be maintained in order to administer drugs extra label to livestock (U.S. Food and Drug All continuous data for pain mitigation treat- Administration, 1994). Thus, use of meloxicam is ment (CONTROL, SHAM, M1, and M2), sam- ELDU and requires a valid VCPR in order to be pling time, and pain mitigation by sampling time administered to livestock in the United States. effects were analyzed using models constructed The plasma meloxicam concentrations of within the MIXED procedure of SAS (SAS calves that were treated with meloxicam are re- Enterprise Guide 4.3, SAS Institute Inc., Cary, NC) ported in Table 1. Plasma from calves that were not in an unpaired-comparison design and mean separ- treated with meloxicam, namely, the CONTROL ation performed with the Tukey’s test designation. and SHAM calves, were not assayed to conserve The daily ambient temperature, relative humidity, resources allocated to this study. The concentra- and wind speed was collected with an on-site wea- tions observed in this study were congruous with ther station (Model # 01604DI, AcuRite Pro Color those reported by Allen et  al. (2013). Allen et  al. Weather Station, Chaney Instrument Co., Lake (2013) indicated some analgesic effects of meloxi- Geneva, WI) and included as covariates nested cam on indicators of pain and inflammation in within calf within day since treatment application 8–10-wk-old Holstein calves that also received within the analysis models. We took a backward a lidocaine cornual nerve block prior to hot iron stepwise approach to analysis output interpretation dehorning. The M1 and M2 treatments resulted in by observing the interaction effect of pain miti- plasma meloxicam concentrations that were not gation treatment and sampling time first. If a sig- different at 24  h following the initial meloxicam nificant interaction effect did not exist, individual dose (P > 0.05). However, the plasma meloxicam main effects (pain mitigation treatment and sam- concentration continued to rise in the M2 treat- pling time, individually) were observed. Significant ment group because a second dose was delivered differences in treatment effects were recognized at to that treatment group at 24 h. For the remainder α ≤ 0.05. of the 96-h plasma sampling period, M2 calves dis- played plasma meloxicam concentrations that were RESULTS AND DISCUSSION approximately twice as elevated as those observed Currently, meloxicam is considered to be a in M1 calves (P < 0.0001). nonsteroidal anti-inflammatory drug (NSAID) be- Table 2 reports the impact of meloxicam treat- longing to the oxicam class that is only approved for ment on growth, algometry, thermography, and se- use in Australia, the European Union, and Canada lect blood constituents of the calves in this study. with a labeled use as therapy for acute respiratory There were no statistically significant treatment ef- disease, diarrhea, mastitis and alleviation of acute fects on average daily gain (ADG), BW, maximum pain and inflammation in cattle (Cotter, 2013; ocular temperature, or plasma cortisol concentra- Health Canada, 2014; Boehringer Ingelheim, 2016; tion. Repenning et al. (2013) also reported that oral European Medicines Agency [EMA], 2018). On meloxicam did not impact ADG during an experi- the contrary, the United States only has one drug, ment involving band castration of beef bulls. Those called flunixin meglumine, approved by the Food results, and ours, concur with Adcock and Tucker Table 1. Least squares means for interaction effect of meloxicam treatment and sampling time on plasma meloxicam concentration (ng/mL) of 3-d-old Holstein calves disbudded with caustic paste (N = 30) Sampling time Meloxicam treatment 24 h 48 h 72 h 96 h Pooled SE P-value (treatment × time) de e f g M1, n = 15 3348.87 3092.07 2608.33 2182.13 156.50 < 0.0001 d a b c M2, n = 15 3570.40 6622.90 5629.86 4872.47 a–g Means with like superscripts within all rows and columns do not differ (P > 0.05). Treatments: 1) M1, caustic paste disbudding and oral Meloxicam (45 mg) with a placebo 24 h later and 2) M2, treatment M1 followed by a second 45-mg dose of meloxicam 24 h later instead of placebo. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Oral meloxicam after caustic paste disbudding 7 Table 2.  Least squares means for the effect of pain treatment options on the growth, algometry, therm- ography, and specific blood constituent concentrations of 3-d-old Holstein calves disbudded with caustic paste (N = 61) Treatment Dependent variable CONTROL (n = 16) SHAM (n = 15) M1 (n = 15) M2 (n = 15) SE P-value Growth ADG, kg 0.3 0.44 0.63 0.53 0.4 0.7445 BW, kg 44.8 44.9 43.4 43.3 1.9 0.3193 Horn bud algometry ab a b ab Mean MNT, kgf 6.57 7.41 6.26 7.20 0.30 0.0212 Ocular thermography Maximum OT, °C 36.21 36.28 35.97 36.03 0.23 0.1281 Blood constituents Cortisol, ng/mL 52.60 57.25 47.59 55.91 6.25 0.2574 ab a b ab Substance P, pg/mL 164.47 198.53 144.50 144.74 12.30 0.0058 a,b Means with like superscripts within each row do not differ (P > 0.05). Treatments: 1) M1, caustic paste disbudding and oral meloxicam (45 mg) with a placebo 24 h later; 2) M2, treatment M1 followed by a second 45-mg dose of meloxicam 24 h later instead of placebo; 3) CONTROL, treatment M1 with placebo in place of meloxicam; and 4) SHAM, sham disbudding with placebo in place of meloxicam. (2018), who reported that the ADG of Holstein and Previous investigations of the relationship between Jersey heifer calves was not impacted by cautery substance P concentration and painful procedures disbudding without pain mitigation at 3 or 35 d of have also reported varying results. A  study that age. These results are similar to another study that included oral meloxicam before and after band investigated pain management associated with horn castration of beef bulls found that plasma con- removal in cattle. Fraccaro et al. (2013) found that centrations of substance P did not differ between meloxicam, along with gabapentin (an antiepileptic castration treatments, whether with or without agent), had no significant influence on blood PGE meloxicam administration (Repenning et al., 2013). concentrations in 6-mo-old Holstein steers that Stock et al. (2015) reported that firocoxib adminis- were provided a lidocaine cornual nerve block and tration to cautery disbudded calves did not influ- mechanically dehorned with a Barnes dehorner. ence plasma substance P concentration compared When substance P is assayed, it is generally re- with a placebo. commended to assay cortisol as well to determine Other investigators have reported stress and if the stressor that impacts substance P release is performance responses that differ from the findings caused by pain or human presence and restraint in this study. Coetzee et  al. (2012) reported that stress. Meléndez et  al. (2018) explained that sub- calves treated with meloxicam for scoop dehorning stance P and cortisol are associated with different reached a level of ADG that was approximately but interrelated mechanisms. Cortisol is a reliable double the ADG of calves that were not treated. indicator of acute stress that returns to normal Heinrich et al. (2009) treated 6–12-wk-old Holstein concentrations following the stress-inducing event, calves that were cautery disbudded with a lidocaine such as an acutely painful event or human presence cornual nerve block and intramuscular meloxicam and restraint. Substance P is a pain-related neuro- (0.5  mg/kg BW). They reported that meloxicam modulator that is implicated in the production of treatment caused a reduction in serum cortisol for prostaglandin E , an inflammatory marker that 6 h following the dehorning procedure but no dif- has been shown to increase after COX enzyme in- ference in concentration was observed 24-h post- hibitors are cleared (Meléndez et al., 2018). In our dehorning (Heinrich et al., 2009). While we cannot study, CONTROL and M2 plasma substance P be certain of all causative factors associated with concentrations were intermediate and not different discrepancies between studies, it is very important (P > 0.05) than SHAM or M1. However, SHAM re- to note the differences in method of administration sulted in greater plasma substance P concentrations (injection vs. oral) and dose (0.5 vs. 1 mg/kg BW) in than M1 (P < 0.0001). It is unclear to us why sub- these studies compared with our study. stance P concentration was elevated for the SHAM Table 3 reports the impact of sampling time on group as this observation does not concur with the the concentration of plasma cortisol and substance findings of other researchers (Coetzee et al., 2012). P in all calves in the study, regardless of treatment. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 8 Karlen et al. Table 3. Least squares means for the effect of sampling time on specific blood constituent concentrations of 3-d-old Holstein calves disbudded with caustic paste (N = 61) Sampling time Dependent variable −5 min 24 h 48 h 72 h 96 h SE P-value Blood constituents a a a b c Cortisol, ng/mL 68.25 58.52 62.68 44.31 32.93 5.89 <0.0001 a b bc bc c Substance P, pg/mL 194.82 172.80 159.60 150.09 137.98 8.36 <0.0001 a–d Means with like superscripts within each row do not differ (P > 0.05). Table 4. Least squares means for the effect of sampling time on growth, algometry, and thermography of 3-d-old Holstein calves disbudded with caustic paste (N = 61) Sampling Time Dependent variable −5 min 0 h 12 h 24 h 48 h 60 h 72 h 96 h 108 h 156 h 204 h 252 h 300 h SE P-value Growth ADG, kg N/A N/A N/A N/A N/A 0.40 N/A N/A 0.60 0.49 0.45 0.46 0.46 0.1 0.0690 d d c c b a e BW, kg N/A N/A 41.2 N/A N/A 42.0 N/A N/A 43.6 44.2 45.1 46.0 46.8 0.1 <0.0001 Algometry ab ab a abcd abc abcd abcd abcd cd abcd bcd d d Mean 8.32 7.91 8.56 7.50 7.72 6.45 6.73 7.03 5.68 6.55 5.99 5.60 5.16 0.50 <0.0001 MNT, kgf Thermography a a ab abc bcd cd d bcd cd cd cd de e Max- 36.97 36.97 36.70 36.54 36.12 36.04 35.75 36.07 35.95 35.99 35.93 35.56 34.99 0.25 <0.0001 imum OT, °C a–e Means with like superscripts within each row do not differ (P > 0.05). The concentration of both molecules in plasma de- first 24 h in particular. However, as the study pro- creased over the course of the sampling period. The gressed and pain status likely shifted from acute to change in cortisol concentration over the course of chronic, our sampling regimen should have been this study is typical of studies where blood sam- sufficient to identify differences in chronic sub- pling occurs at multiple time points over several stance P production in response to the disbudding hours to multiple days. It is generally acknowledged lesions. that cortisol concentration typically increases rap- Table 4 shows the impact of sampling time on idly following exposure to acute stressors and ADG, BW, MNT, and   ocular temperature (OT). then declines toward normal concentrations that ADG was not significantly different between sam- vary rhythmically throughout the day, even with pling times (P  =  0.74). However, BW increased the prolonged presence of the same acute stressor across sampling times (P  <  0.0001). Both MNT (Apple et al., 2005). Stock et al. (2015) identified a and OT declined over the sampling period. It is similar time effect on cortisol concentrations in pre- possible that this may have been a result of the weaned dairy calves that were cautery disbudded. adjustment of homeostatic mechanisms in the Our data suggest that the change in substance P neonatal calves following parturition. In a re- and cortisol concentration across all calves over view, Kirovski (2015) described the dependence time may have been resultant of human presence of neonatal calves on nonshivering thermogen- and restraint stress. Investigation of this concept is esis, a process that involves the uncoupling of warranted and necessary. In addition, specific guid- adenosine triphosphate (ATP) synthase from the ance regarding the optimal time of day to measure electron transport chain, resulting in heat produc- substance P is needed for cattle. In our study, we tion in brown adipose tissue. Brown adipose tissue consistently collected all blood for assay during the recedes during the first month of life for bovine same time frame (0500–0700 hours) each day. It is calves and other neonatal ruminants. It is possible possible that the 24-h time lapse between substance that the decline in OT that we observed was an in- P sample collections in our study could have missed dication of the progressive reduction in nonshiver- changes in concentration that occurred during the ing thermogenesis through the first month of life. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Oral meloxicam after caustic paste disbudding 9 Table 5. Least squares means for the interaction effect of meloxicam treatment and sampling time on mean horn bud temperature and mean horn bud to ocular temperature ratio of 3-d-old Holstein calves disbudded with caustic paste (N = 61) Sampling time Dependent variable −5 min 0 h 12 h 24 h 48 h 60 h 72 h 96 h 108 h 156 h 204 h 252 h 300 h SE P-value Mean horn bud temperature, °C CONTROL 25.46 36.75 33.55 32.49 32.29 32.86 32.93 33.60 33.66 33.93 34.48 34.02 33.46 0.46 <0.0001 SHAM 25.99 34.57 33.46 33.46 32.13 33.20 32.19 32.94 33.30 33.28 33.40 33.28 33.22 M1 25.19 36.73 33.01 32.76 33.14 33.20 32.79 32.87 33.25 33.60 33.69 33.43 32.93 ab M2 23.96 36.64 33.01 32.14 33.41 33.29 32.60 33.15 33.00 33.45 33.09 33.93 33.21 Mean horn bud: ocular temperature CONTROL 0.68 1.00 0.91 0.89 0.90 0.91 0.92 0.93 0.93 0.94 0.95 0.96 0.96 0.01 <0.0001 SHAM 0.69 0.93 0.91 0.92 0.89 0.92 0.90 0.91 0.92 0.91 0.92 0.93 0.93 M1 0.67 1.00 0.90 0.89 0.91 0.93 0.92 0.91 0.93 0.94 0.95 0.94 0.96 M2 0.65 1.00 0.90 0.88 0.92 0.92 0.92 0.92 0.92 0.94 0.93 0.96 0.95 a,b Means within dependent variable with like superscripts within column do not differ (P > 0.05). Treatments: 1) M1, caustic paste disbudding and oral meloxicam (45 mg) with a placebo 24 h later; 2) M2, treatment M1 followed by a second 45-mg dose of meloxicam 24 h later instead of placebo; 3) CONTROL, treatment M1 with placebo in place of meloxicam; and 4) SHAM, sham disbudding with placebo in place of meloxicam. Glynn et  al. (2013) reported a similar time effect of nondisbudded tissue during a 3-wk period fol- on MNT values proximal to the horns of Holstein lowing disbudding. However, the same project steers. found that calves remained sensitive to touch near Table 5 reports the interaction effects of meloxi- the disbudding site for 9 wk. Overall, our findings cam treatment and sampling time on mean horn align with the conclusions of Coetzee et al. (2012) bud temperature by thermography and mean horn that administration of meloxicam as the only pain bud to ocular temperature ratio. It is important to mitigation therapy for horn tissue removal is insuf- note that the values reported at −5  min for both ficient in the control of acute pain but may be valu- variables occurred before the horn buds were shaved able in the control of chronic pain associated with for paste application. Longer and thicker hair pro- postprocedure healing. It is important to note that vided greater insulation and cooler temperatures at the discussion of the results in this study included the surface of the hair coat that was captured in the reference to an array of studies that focused on the thermal image. Both dependent variables exhibited responses of cattle to horn removal and the im- the treatment by sampling time interaction effect at pact of multiple pain mitigation strategies on those time 0 h, which was immediately after the applica- responses. Due to the limited amount of data re- tion of caustic paste for CONTROL, M1, and M2 garding caustic paste disbudding that was available calves or the noncaustic analog for SHAM. Horn at the time of this publication, reference to other bud temperature was greater in the CONTROL and disbudding methods was necessary. However, the M1 groups than SHAM but not different than M2. specific mechanisms of action are vastly different Mean horn bud to ocular temperature ratio was between cautery, caustic, and mechanical horn re- less in the SHAM group than any treatment that moval and different mechanisms and magnitudes of received caustic paste. No other difference was ob- pain response must be expected and acknowledged. served between treatments at any other sampling Our data suggest that meloxicam, as applied in time. The data for both horn bud temperature and our study, is not sufficient as the primary therapy horn bud to ocular temperature ratio suggest that to control pain and inflammation associated with the heat production associated with apparent tissue caustic paste disbudding. Further study regarding inflammation from caustic paste application did the impact of cornual nerve block in conjunction not last longer than 12 h, regardless of meloxicam with meloxicam administration on caustic paste treatment. It is important to note that the absence disbudded calves is warranted as other previously of heat is not sufficient to conclude that pain has cited work has identified the value of regional anes- subsided. Adcock and Tucker (2018) found that thesia to controlling the acute pain associated with the surface temperature of cautery disbudding caustic paste disbudding and the need for effective wounds did not differ from the surface temperature chronic pain mitigation. The variables that we Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 10 Karlen et al. Coetzee,  J.  F., R.  A.  Mosher, B.  KuKanich, R.  Gehring, utilized in this study may not have been completely B.  Robert, J.  B.  Reinbold, and B.  J.  White. 2012. effective in quantifying the magnitude of chronic Pharmacokinetics and effect of intravenous meloxi- pain associated with caustic paste administra- cam in weaned Holstein calves following scoop dehorn- tion as no overall effect was observed between the ing without local anesthesia. BMC Vet. Res. 8:153. CONTROL, M1, and M2 treatment groups over doi:10.1186/1746-6148-8-153 Cotter,  F . 2013. A GLP tissue residue depletion study in calves fol- time. This study would have benefitted from inclu- lowing oral / buccal administration of meloxicam. Available sion of assays of behavioral indicators of pain and from https://www.mla.com.au/research-and-development/ discomfort. Preliminary work in our lab has sug- search-rd-reports/final-report-details/Animal-Welfare/A- gested that behaviors associated with chronic pain GLP-tissue-residue-depletion-in-calves-following-oral- and discomfort may be subtle in young calves and buccal-administration-of-meloxicam/2072#. Accessed considerable care must be taken to quantify behav- May 31, 2019. European Medicines Agency (EMA). 2018. Metacam (meloxi- iors that effectively represent changes in pain status. cam): An overview of Metacam and why it is authorized in A behavioral assay with capability to detect pain-in- the EU. Available from https://www.ema.europa.eu/docu- duced pessimism and anhedonia has recently been ments/overview/metacam-epar-summary-public_en.pdf. suggested by other researchers for the assessment Accessed February 25, 2019. of mood changes in calves that were cautery dis- Fraccaro, E., J. F. Coetzee, R. Odore, L. N. Edwards-Callaway, B.  Kukanich, P.  Badino, L.  Bertolotti, H.  Glynn, budded (Lecorps et al., 2019). We recommend that J. Dockweiler, K. Allen, et al. 2013. A study to compare future work to investigate the impact of pain miti- circulating flunixin, meloxicam and gabapentin concen- gation strategies on pain response in calves, specif- trations with prostaglandin E₂ levels in calves under- ically, and animals, in general, include variables that going dehorning. Res. Vet. Sci. 95:204–211. doi:10.1016/j. effectively quantify both physiological and behav- rvsc.2013.01.018 ioral indicators of changes to affective states. 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Dairy Sci. caustic paste after non-steroidal-anti-inflammatory 100:6429–6441. doi:10.3168/jds.2017-12724 Translate basic science to industry innovation http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Translational Animal Science Oxford University Press

Efficacy of oral meloxicam as primary pain mitigation following caustic paste disbudding of three day old Holstein calves

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Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Efficacy of oral meloxicam as primary pain mitigation following caustic paste disbudding of three day old Holstein calves † ‡ || $ ¶, Kyle J. Karlen, Faith S. Baier, Sara L. Odegard, Ruth M. Baumann, Johann F. Coetzee, || ||,2 Sylvia I. Kehoe, and Kurt D. Vogel † ‡ School of Veterinary Medicine, University of Wisconsin, Madison, WI 53706; Department of Animal Sciences, || Colorado State University, Fort Collins, CO 80523; Department of Animal and Food Science, University of $ ¶ Wisconsin, River Falls, WI 54022; Golden Eagle Lodge, Grand Marais, MN 55604; and College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506 ABSTRACT: The purpose of this study was to in- Plasma was collected and analyzed for cortisol and vestigate the effects of perioperative administration substance P concentrations. Substance P and cor- of oral meloxicam prior to and following the ap- tisol decreased in all animals over time, regardless plication of caustic paste to disbud neonatal dairy of treatment. Mean plasma substance P concentra- calves. Sixty-one 3–4-d-old Holstein heifer calves tion across all time points was greater (P  <  0.05) were randomly assigned to one of four treatment in the SHAM group than M1 or M2 but not dif- groups of 15–16 calves. The treatment groups were: ferent (P > 0.05) than the CONTROL group. The 1) M1, caustic paste disbudding and oral meloxicam MNT and ocular temperatures decreased over time (45 mg) with a placebo 24 h later; 2) M2, treatment across all treatments (P  <  0.05). Mean BW in- M1 followed by a second 45-mg dose of meloxicam creased over time across all treatments (P < 0.05). 24 h later instead of placebo; 3) CONTROL, treat- A significant interaction (P  < 0.05) between treat- ment M1 with placebo in place of meloxicam; and ment and sampling time was observed at 12 h fol- 4) SHAM, sham disbudding with placebo in place lowing treatment application for both mean horn of meloxicam. Infrared thermography was used to bud temperature and the ratio between horn bud quantify eye and horn bud temperatures. Pressure and ocular temperature. Overall, the results of this algometry was used to measure Mechanical noci- study suggest that meloxicam administration at a ceptive threshold (MNT) surrounding the horn dose of 45 mg per animal may have limited influ- bud. Average daily gain and body weight (BW) ence as the primary modulator of pain and inflam- were obtained by weighing each animal throughout matory response in calves that have been disbudded the study and calculating the changes over time. with caustic paste at 3 d of age. Key words: calf, dehorn, disbud, horn, meloxicam, pain © The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. Transl. Anim. Sci. 2019.XX:0-0 doi: 10.1093/tas/txz151 The authors greatly appreciated the hospitality of the com- thoughtful and valuable input regarding the content of our mercial dairy that allowed us to access and use their calves for manuscript. This project was funded by the University of Wis- this study. We would also like to thank E.  Andersen, A.  Bue, consin—River Falls (UWRF) Undergraduate Research, Schol- K. Guden, S. Heck, J. Hollatz, A. Keller, A. Long, C. McClin- arly, and Creative Activity program, UWRF Falcon Scholars tock, K. Schiller, and J. Spoehr for their assistance in sample col- program, and a generous gift from The Kraft Heinz Company. lection and management. We also appreciated the guidance and Corresponding author: kurt.vogel@uwrf.edu assistance of J. Peterson from the Iowa State University PHAST Received June 29, 2019. Lab. In addition, we thank the anonymous reviewers for their Accepted October 1, 2019. 1 Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 2 Karlen et al. INTRODUCTION One potential option for pain mitigation for caustic paste disbudding is meloxicam. Meloxicam One of the most common painful procedures is a long-acting NSAID that is similarly available for North American dairy cattle is removal of both parenterally and orally, has peak availability the horns or horn buds. In 2014, 94.3% of North at approximately 12  h, and has a half-life of 28  h American dairy operations reported routine re- (Coetzee et  al., 2009). This makes meloxicam moval of the horns of heifer calves on their prem- uniquely suited for administration up to 8  h prior ises (National Animal Health Monitoring System to dehorning or shortly before the application of [NAHMS], 2014). Of those dairy operations, 16.4% caustic paste and may make a second dose unneces- utilized caustic paste for disbudding at 2.3  wk of sary until days after the initial dose was adminis- age (NAHMS, 2014). In the same survey, 5.6% of tered as meloxicam may suppress the concentrations the dairy operations that used caustic paste for rou- of prostaglandin E2 (PGE ), an important pros- tine disbudding of dairy calves utilized analgesic taglandin associated with the Cyclooxygenase-2 and/or anesthetic pain mitigation (NAHMS, 2014). (COX-2) inflammatory pathway, for as long as Across all dairy operations that performed routine 3 d (Allen et  al., 2013). To this point in time, no horn removal, pain mitigation use was dependent previous work had been published regarding the on the person that performed the procedure as impact of meloxicam as the primary pain mitiga- 62.7% of veterinarians, 29.2% of employees, and tion therapy for caustic paste disbudding, and the 14.9% of owner/operators reported that they used research on caustic paste disbudding in general is anesthetic and/or analgesic pain mitigation for any limited. As a result, this study was focused on ex- routine horn removal procedures (NAHMS, 2014). ploring the impact of practical meloxicam admin- Caustic paste disbudding has been suggested to istration techniques on dairy calves. The objective cause less acute pain response than cautery disbud- of this study was to investigate the effects of peri- ding (Vickers et al., 2005), and the procedure itself operative administration of oral meloxicam prior has a more innocuous appearance than other horn to and following the application of caustic paste to removal methods (Vickers et  al., 2005). However, disbud neonatal dairy calves. more recent work has suggested that caustic paste may be more noxious than cautery disbudding MATERIALS AND METHODS but deceiving to assess due to the differing meth- ods of application and action (Braz et  al., 2012; Institutional Animal Care and Use Committee Hempstead et al., 2018). Approval Based on behavioral and cortisol responses, peak pain appears to occur within the first 4 h fol- Prior to the commencement of this study, animal lowing caustic paste application, with the poten- use and associated procedures were approved by the tial for chronic pain after that time period (Morisse University of Wisconsin—River Falls Institutional et al., 1995; Stilwell et al., 2009). Winder et al. (2017) Animal Care and Use Committee. The protocol reported that acute pain persisted for a minimum of number associated with this study was A15-16–7. 180 min following the application of caustic paste to disbud 3–5-d-old Holstein calves that were treated Animals and Housing with 0.5 mg/kg subcutaneous meloxicam 15 min be- fore caustic paste application. In the same study, re- Sixty-one female Holstein calves (41.2 ± 0.1 kg gional anesthesia in the form of a lidocaine cornual body weight [BW]) were housed in individual out- nerve block (5 mL per side) effectively reduced acute door hutches on a sand base with a layer of wood pain indication by algometry for approximately shavings and straw bedding inside each hutch (ap- 180  min in calves that were treated with the same proximate area: 2.4 m ) and in an individual out- dose of meloxicam 15 min before caustic paste appli- door exercise space accompanying each hutch cation. Braz et al. (2012) indicated that caustic paste (approximate area: 2 m ). Each calf was fitted disbudding caused severe pain for the first 30 min fol- with a thermal jacket that was worn for the dur- lowing paste application and found tramadol to be ation of the study at the time of separation from ineffective in controlling acute pain associated with her dam. All calves were born and housed on the the procedure. Stilwell et  al. (2008, 2009) reported same commercial dairy farm in Western Wisconsin that flunixin meglumine alone was also ineffective between February and April 2016. Each calf was in controlling a postapplication cortisol increase in separated from her dam within an hour of birth dairy calves following caustic paste application. and fed 3.8 liters of colostrum by gastroesophageal Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Oral meloxicam after caustic paste disbudding 3 tube within 2 h of birth. A second feeding of col- Administration of Oral Meloxicam ostrum was administered by bottle 12–24  h after All calves in this experiment were fed 1.9 li- the initial colostrum feeding. Following the second ters of pasteurized milk after the administration colostrum feeding, each calf was fed 1.9 liters of of meloxicam and collection of measurements at pasteurized whole milk three times daily. All calves each time point. The experimental dose of meloxi- were enrolled in the study and received their as- cam or placebo was administered via balling signed treatments at exactly 3 d of age. The number gun. Three—15  mg meloxicam tablets (Unichem of calves that were enrolled in the study on each Pharmaceuticals (USA) Inc., Rochelle Park, NJ) day varied depending on the number of calves that were loaded into empty gel capsules (size 00, Torpac were born 3 d earlier. The treatments were assigned Inc., Fairfield, NJ) and two capsules at a time were in rolling order such that each treatment was repre- administered to calves in the M1 and M2 treatment sented once in each group of four calves that were groups at the scheduled administration times. Of consecutively enrolled in the study. The specific the two capsules, one capsule was always empty treatment that each calf received was dependent on and served to assist in holding the meloxicam or the chronological birth order and next treatment placebo capsule in the balling gun during passage assignment in the repeating treatment applica- into the esophagus. The second capsule contained a tion order. Farm staff were blind to all treatments. total of 45 mg of meloxicam. Each dose of meloxi- Some project personnel were not blind to the treat- cam provided approximately 1.0 mg meloxicam per ment groups if they were involved in treatment ap- kg of calf BW. Meloxicam or placebo delivery oc- plication. However, the specific outcomes that were curred approximately 5 min before the disbudding assessed were unlikely to be affected by the treat- procedure and just before the feeding 24 h postdis- ment knowledge of those personnel. budding. Calves that received placebo at either or both time points received empty gel capsules via Experimental Design and Treatments balling gun instead of meloxicam in gel capsules. Each calf was assigned to one of four treat- ment groups. Treatment assignments were pro- Disbudding and Sham Disbudding Procedure vided in rolling order as calves were enrolled in the study at 3 d of age. The treatment groups Prior to disbudding treatment application, a were: 1)  M1, caustic paste disbudding and oral battery-operated hair clipper was used to trim the meloxicam (45  mg) with a placebo 24  h later; hair to a length of approximately 1  mm within a 2) M2, treatment M1 followed by a second 45-mg 3-cm radius of the horn buds. A  commercially dose of meloxicam 24 h later instead of placebo; available caustic paste product that contained a 3)  CONTROL, treatment M1 with placebo in mixture of sodium and calcium hydroxide (Dr place of meloxicam; and 4)  SHAM, sham dis- Naylor Dehorning Paste, H.  W. Naylor Company budding with placebo in place of meloxicam. Inc, Morris, NY) or a noncaustic analog (Up and The treatment application and sample collection Up creamy diaper rash ointment, Target Stores, timeline may be observed in Figure 1. At each Minneapolis, MN) was administered to the horn time point, ocular and horn bud thermographs bud. The noncaustic analog was similar in consist- were collected followed by algometry data for the ency to the caustic paste but differed in color. The tissue surrounding the horn buds. On odd-num- caustic paste was light red in color and the non- bered evenings beginning on day 1, BW was also caustic analog was white. Paste, either caustic or recorded in addition to the thermographs and noncaustic, was evenly applied with a gloved hand algometry readings. Thermographs and algom- at a thickness of approximately 1  mm to an area etry readings were collected during the morning on the horn bud that was approximately 18 mm in feedings on days 1–5 and on odd-numbered even- diameter ings for each calf, resulting in measurements for time points −5 min, 0, 12, 24, 48, 60, 72, 96, 108, BW and Average Daily Gain 156, 204, 252, and 300 h from disbudding. For all treatments and measurements except blood col- The BW of each animal involved in this study lection, calves were restrained in sternal recum- was collected from a digital hanging scale (Product bency with one student straddling the calf while #34626, Roughneck Digital Hanging Scale, applying one hand to the base of the head and one Northern Tool + Equipment, Burnsville, MN) to the nose to limit movement of the calf ’s head. equipped with a commercially available sling (Item Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 4 Karlen et al. Caustic Paste applied to M1, M2, and CONTROL. Non-caustic analog applied for SHAM. Meloxicam applied to M2. Meloxicam applied to M1 Placebo and M2. applied to CONTROL, Placebo applied SHAM, and to CONTROL M1. and SHAM. BD BD BD BD BD TH TH TH TH TH TH TH TH TH TH TH TH TH PA PA PA PA PA PA PA PA PA PA PA PA PA BW BW BW BW BW BW BW BW BW BW BW BW BW - 0.5 0 12 24 48 60 72 108 156 204 252 300 Hours Figure 1. This timeline graphic outlines the treatment administration times and sample collection times over the course of the study. The treat- ment groups were: 1) M1: caustic paste disbudding and oral meloxicam (45 mg) with a placebo 24 h later; 2) M2: treatment M1 followed by a second 45-mg dose of meloxicam 24 h later instead of placebo; 3) CONTROL: treatment M1 with placebo in place of meloxicam; and 4) SHAM: sham disbudding with placebo in place of meloxicam. Sample collections are defined as: BD = blood draw, TH = ocular and horn bud infrared therm- ography, PA = pressure algometry, WT = body weight. Time point 0 marks the measurements recorded immediately after treatment application. # 17123, Calf Weighing Sling, Valley Vet Supply, of collection, all whole blood was centrifuged in Marysville, KS) to safely suspend each calf during the original collection tube for cortisol and the so- weight collection. BW was recorded on odd-num- dium EDTA plus benzamidine tubes for substance bered evenings over a 13-d period beginning on the P at 2,500 × g for 15 min. Plasma was transferred day of disbudding and meloxicam treatment ap- to microcentrifuge tubes with disposable transfer plication. To calculate ADG, the total amount of pipettes and then frozen in liquid nitrogen before BW change between BW collection events was div- transferring to storage at −80 °C until analysis via ided by the total number of experimental days that radioimmunoassay at the Iowa State Veterinary elapsed since the previous BW collection event. diagnostic laboratory. Blood Collection Cortisol and Substance P Quantification Whole blood was collected via jugular veni- Plasma substance P and cortisol concentra- puncture with 21-g × 2.54-cm blood collection tions were quantified at the Iowa State University needles (REF 450042, Greiner Bio-One GmbH, Veterinary Diagnostic Laboratory following pro- Kremsmünster, Austria) from all animals in each cedures that were described by Kleinhenz et  al. of the four treatment groups. Blood was collected (2018). A  commercially available kit (catalog # prior to the administration of meloxicam and 06B-256440, Corti-Cote Cortisol RIA Kit, MP caustic paste (time −5 min) to obtain baseline sub- Biomedicals Inc., Santa Ana, CA) was used to es- stance P and cortisol concentrations, in addition to timate plasma cortisol concentrations for each times 24, 48, 72, and 96 h. At each collection time, sample in duplicate following the manufacturer’s a single 10-mL nonadditive blood collection tube procedure. The duplicate values were averaged for (BD, Franklin Lakes, NJ) and a single 6-mL blood each sample prior to statistical analysis. The coef- collection tube with lithium heparin additive (BD, ficient of variation for intra-assay variability was Franklin Lakes, NJ) were collected from each calf. 10.31%. The coefficient of variation for interassay Immediately after collection, 6 mL of whole blood variability was 5.59%. from the nonadditive tube was transferred to a 6-mL Substance P was quantified in nonextracted blood collection tube that contained sodium ethyl- plasma following procedures that were out- enediaminetetraacetic acid (EDTA) additive plus lined by Kleinhenz et  al. (2018), Van Engen et  al. 300  μL of benzamidine solution. Within 30  min (2014), and Liu et  al. (2008). A  two-antibody Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Oral meloxicam after caustic paste disbudding 5 radioimmunoassay (RIA) was used to estimate Kleinhenz et al., 2018). An infrared camera (FLIR plasma substance P concentrations for each sample E8 Thermal Camera, FLIR Systems, Boston, MA) in duplicate. The duplicate values were averaged for was used to collect three thermal images of each each sample prior to statistical analysis. The coef- eye and each horn bud for each animal at the time ficient of variation for intra-assay variability was points identified in Figure 1. Thermographs were 10.55%. The coefficient of variation for interassay collected from a distance of approximately 30.5 cm variability was 27.64%. with the camera focal point in perpendicular align- ment with the target location. A laser tape measure (Bosch GLM 10 Compact Laser Measure, Robert Meloxicam Quantification Bosch GmbH, Stuttgart, Germany) was attached Plasma meloxicam concentrations were quanti- to the thermal camera to facilitate rapid, accurate, fied for calves in the M1 and M2 treatment groups and consistent distance determination. at the Iowa State University Veterinary Diagnostic Ocular thermographs were collected first and Laboratory following procedures similar to those then each calf was blindfolded and allowed to ac- described by Repenning et al. (2013). High-pressure climate to the blindfold while horn bud thermo- liquid chromatography tandem–mass spectrom- graphs were collected prior to pressure algometry. etry (HPLC–MS) was performed with a LTQ ion Stock et  al. (2015) also used a blindfold to assist trap (Thermo Scientific, San Jose, CA) coupled in maintaining stable body and head position dur- with an Agilent 1100 series pump and autosampler ing algometry. Thermographs were analyzed with (Agilent Technologies, Santa Clara, CA) to ana- FLIR Tools software (FLIR Systems, Boston, lyze each sample in single. The instrument injec- MA). For each image, the area of interest (ei- tion volume was set to 20 μL. The mobile phases ther eye or horn bud) was selected via a function were A: 0.1% formic acid in water; and B: 0.1% in FLIR Tools, and the maximum temperature of formic acid in acetonitrile. The gradient curve the selected field was recorded and entered into a began at 25% B with a linear gradient to 95% B in Microsoft Excel spreadsheet and used for statistical 5 min, maintained for 1.75 min, then re-equilibra- analysis. The mean of the three maximum temper- tion to 25% B. The o fl w rate was 0.25 mL/min and atures recorded from the thermal images for each increased to 0.325 mL/min at 5.25 min. Separation eye and each horn bud was ultimately included in was performed with an UltraCore C18 100- × the statistical analyses. 2.1-mm column (Kinetex XB-C18, Phenomenex, Totrrence, CA). Meloxicam eluted at 5.4 min and Pressure Algometry piroxicam eluted at 4.7  min. Four selected reac- tion monitoring transitions each were monitored Pressure algometry has been used to assess the for meloxicam (115, 141, 184, and 352 m/z) and mechanical nociceptive threshold (MNT) of ani- piroxicam (227, 270, 282, and 332 m/z). Sequences mals in previous experiments investigating the pain were batch processed with a method developed in response to disbudding (Heinrich et al., 2010; Allen Xcalibur software (Thermo Scientific, San Jose, et al., 2013; Glynn et al., 2013; Winder et al., 2017; CA). The method automatically identified and in- Adcock and Tucker, 2018). MNT has been used tegrated each peak in each sample and calculated as a measure of sensitivity to physical stimulus; a the calibration curve based on a weighted (1/X) greater MNT value suggests a greater tolerance to quadratic fit. Plasma concentrations of meloxi- force and decreased sensitivity. A  pressure algom- cam in unknown samples were calculated by the eter (Wagner Force Ten FDX 25 Compact Digital Xcalibur software based on the calibration curve. Force Gage, Wagner Instruments, CT) was used The correlation coefficient (r ) was > 0.98 across to apply pressure to three areas anterior to, lateral the concentration range of 5–5,000  ng/mL. QC to, and posterior to each horn bud at the edge of samples were within tolerance of ±15% of the the paste that was applied. A 1-cm flat rubber tip nominal value. accessory was affixed to the algometer. Force was applied at an approximate rate of 1.0  kgf/s until the animal attempted to move its head away from Infrared Thermography the applied pressure, similar to the procedure in Infrared thermography has previously been used Winder et  al. (2017). Maximum force did not ex- to observe the change in temperature of the eye dur- ceed 10 kg force to prevent excessive discomfort for ing painful procedures in several studies (Stewart the calf. The maximum force applied prior to move- et  al., 2008; Allen et  al., 2013; Stock et  al., 2015; ment away from the pressure was recorded for each Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 6 Karlen et al. of the three locations for each of the horn buds and Drug Administration (FDA) as an NSAID at each collection time identified in Figure 1. The for cattle with a label referencing pain treatment. mean MNT for each horn bud at each collection Therefore, any administration of meloxicam falls time was included in the statistical analyses. into the category of extra label drug use (ELDU). A  valid veterinary patient client relationship Statistical Analyses (VCPR) must be maintained in order to administer drugs extra label to livestock (U.S. Food and Drug All continuous data for pain mitigation treat- Administration, 1994). Thus, use of meloxicam is ment (CONTROL, SHAM, M1, and M2), sam- ELDU and requires a valid VCPR in order to be pling time, and pain mitigation by sampling time administered to livestock in the United States. effects were analyzed using models constructed The plasma meloxicam concentrations of within the MIXED procedure of SAS (SAS calves that were treated with meloxicam are re- Enterprise Guide 4.3, SAS Institute Inc., Cary, NC) ported in Table 1. Plasma from calves that were not in an unpaired-comparison design and mean separ- treated with meloxicam, namely, the CONTROL ation performed with the Tukey’s test designation. and SHAM calves, were not assayed to conserve The daily ambient temperature, relative humidity, resources allocated to this study. The concentra- and wind speed was collected with an on-site wea- tions observed in this study were congruous with ther station (Model # 01604DI, AcuRite Pro Color those reported by Allen et  al. (2013). Allen et  al. Weather Station, Chaney Instrument Co., Lake (2013) indicated some analgesic effects of meloxi- Geneva, WI) and included as covariates nested cam on indicators of pain and inflammation in within calf within day since treatment application 8–10-wk-old Holstein calves that also received within the analysis models. We took a backward a lidocaine cornual nerve block prior to hot iron stepwise approach to analysis output interpretation dehorning. The M1 and M2 treatments resulted in by observing the interaction effect of pain miti- plasma meloxicam concentrations that were not gation treatment and sampling time first. If a sig- different at 24  h following the initial meloxicam nificant interaction effect did not exist, individual dose (P > 0.05). However, the plasma meloxicam main effects (pain mitigation treatment and sam- concentration continued to rise in the M2 treat- pling time, individually) were observed. Significant ment group because a second dose was delivered differences in treatment effects were recognized at to that treatment group at 24 h. For the remainder α ≤ 0.05. of the 96-h plasma sampling period, M2 calves dis- played plasma meloxicam concentrations that were RESULTS AND DISCUSSION approximately twice as elevated as those observed Currently, meloxicam is considered to be a in M1 calves (P < 0.0001). nonsteroidal anti-inflammatory drug (NSAID) be- Table 2 reports the impact of meloxicam treat- longing to the oxicam class that is only approved for ment on growth, algometry, thermography, and se- use in Australia, the European Union, and Canada lect blood constituents of the calves in this study. with a labeled use as therapy for acute respiratory There were no statistically significant treatment ef- disease, diarrhea, mastitis and alleviation of acute fects on average daily gain (ADG), BW, maximum pain and inflammation in cattle (Cotter, 2013; ocular temperature, or plasma cortisol concentra- Health Canada, 2014; Boehringer Ingelheim, 2016; tion. Repenning et al. (2013) also reported that oral European Medicines Agency [EMA], 2018). On meloxicam did not impact ADG during an experi- the contrary, the United States only has one drug, ment involving band castration of beef bulls. Those called flunixin meglumine, approved by the Food results, and ours, concur with Adcock and Tucker Table 1. Least squares means for interaction effect of meloxicam treatment and sampling time on plasma meloxicam concentration (ng/mL) of 3-d-old Holstein calves disbudded with caustic paste (N = 30) Sampling time Meloxicam treatment 24 h 48 h 72 h 96 h Pooled SE P-value (treatment × time) de e f g M1, n = 15 3348.87 3092.07 2608.33 2182.13 156.50 < 0.0001 d a b c M2, n = 15 3570.40 6622.90 5629.86 4872.47 a–g Means with like superscripts within all rows and columns do not differ (P > 0.05). Treatments: 1) M1, caustic paste disbudding and oral Meloxicam (45 mg) with a placebo 24 h later and 2) M2, treatment M1 followed by a second 45-mg dose of meloxicam 24 h later instead of placebo. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Oral meloxicam after caustic paste disbudding 7 Table 2.  Least squares means for the effect of pain treatment options on the growth, algometry, therm- ography, and specific blood constituent concentrations of 3-d-old Holstein calves disbudded with caustic paste (N = 61) Treatment Dependent variable CONTROL (n = 16) SHAM (n = 15) M1 (n = 15) M2 (n = 15) SE P-value Growth ADG, kg 0.3 0.44 0.63 0.53 0.4 0.7445 BW, kg 44.8 44.9 43.4 43.3 1.9 0.3193 Horn bud algometry ab a b ab Mean MNT, kgf 6.57 7.41 6.26 7.20 0.30 0.0212 Ocular thermography Maximum OT, °C 36.21 36.28 35.97 36.03 0.23 0.1281 Blood constituents Cortisol, ng/mL 52.60 57.25 47.59 55.91 6.25 0.2574 ab a b ab Substance P, pg/mL 164.47 198.53 144.50 144.74 12.30 0.0058 a,b Means with like superscripts within each row do not differ (P > 0.05). Treatments: 1) M1, caustic paste disbudding and oral meloxicam (45 mg) with a placebo 24 h later; 2) M2, treatment M1 followed by a second 45-mg dose of meloxicam 24 h later instead of placebo; 3) CONTROL, treatment M1 with placebo in place of meloxicam; and 4) SHAM, sham disbudding with placebo in place of meloxicam. (2018), who reported that the ADG of Holstein and Previous investigations of the relationship between Jersey heifer calves was not impacted by cautery substance P concentration and painful procedures disbudding without pain mitigation at 3 or 35 d of have also reported varying results. A  study that age. These results are similar to another study that included oral meloxicam before and after band investigated pain management associated with horn castration of beef bulls found that plasma con- removal in cattle. Fraccaro et al. (2013) found that centrations of substance P did not differ between meloxicam, along with gabapentin (an antiepileptic castration treatments, whether with or without agent), had no significant influence on blood PGE meloxicam administration (Repenning et al., 2013). concentrations in 6-mo-old Holstein steers that Stock et al. (2015) reported that firocoxib adminis- were provided a lidocaine cornual nerve block and tration to cautery disbudded calves did not influ- mechanically dehorned with a Barnes dehorner. ence plasma substance P concentration compared When substance P is assayed, it is generally re- with a placebo. commended to assay cortisol as well to determine Other investigators have reported stress and if the stressor that impacts substance P release is performance responses that differ from the findings caused by pain or human presence and restraint in this study. Coetzee et  al. (2012) reported that stress. Meléndez et  al. (2018) explained that sub- calves treated with meloxicam for scoop dehorning stance P and cortisol are associated with different reached a level of ADG that was approximately but interrelated mechanisms. Cortisol is a reliable double the ADG of calves that were not treated. indicator of acute stress that returns to normal Heinrich et al. (2009) treated 6–12-wk-old Holstein concentrations following the stress-inducing event, calves that were cautery disbudded with a lidocaine such as an acutely painful event or human presence cornual nerve block and intramuscular meloxicam and restraint. Substance P is a pain-related neuro- (0.5  mg/kg BW). They reported that meloxicam modulator that is implicated in the production of treatment caused a reduction in serum cortisol for prostaglandin E , an inflammatory marker that 6 h following the dehorning procedure but no dif- has been shown to increase after COX enzyme in- ference in concentration was observed 24-h post- hibitors are cleared (Meléndez et al., 2018). In our dehorning (Heinrich et al., 2009). While we cannot study, CONTROL and M2 plasma substance P be certain of all causative factors associated with concentrations were intermediate and not different discrepancies between studies, it is very important (P > 0.05) than SHAM or M1. However, SHAM re- to note the differences in method of administration sulted in greater plasma substance P concentrations (injection vs. oral) and dose (0.5 vs. 1 mg/kg BW) in than M1 (P < 0.0001). It is unclear to us why sub- these studies compared with our study. stance P concentration was elevated for the SHAM Table 3 reports the impact of sampling time on group as this observation does not concur with the the concentration of plasma cortisol and substance findings of other researchers (Coetzee et al., 2012). P in all calves in the study, regardless of treatment. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 8 Karlen et al. Table 3. Least squares means for the effect of sampling time on specific blood constituent concentrations of 3-d-old Holstein calves disbudded with caustic paste (N = 61) Sampling time Dependent variable −5 min 24 h 48 h 72 h 96 h SE P-value Blood constituents a a a b c Cortisol, ng/mL 68.25 58.52 62.68 44.31 32.93 5.89 <0.0001 a b bc bc c Substance P, pg/mL 194.82 172.80 159.60 150.09 137.98 8.36 <0.0001 a–d Means with like superscripts within each row do not differ (P > 0.05). Table 4. Least squares means for the effect of sampling time on growth, algometry, and thermography of 3-d-old Holstein calves disbudded with caustic paste (N = 61) Sampling Time Dependent variable −5 min 0 h 12 h 24 h 48 h 60 h 72 h 96 h 108 h 156 h 204 h 252 h 300 h SE P-value Growth ADG, kg N/A N/A N/A N/A N/A 0.40 N/A N/A 0.60 0.49 0.45 0.46 0.46 0.1 0.0690 d d c c b a e BW, kg N/A N/A 41.2 N/A N/A 42.0 N/A N/A 43.6 44.2 45.1 46.0 46.8 0.1 <0.0001 Algometry ab ab a abcd abc abcd abcd abcd cd abcd bcd d d Mean 8.32 7.91 8.56 7.50 7.72 6.45 6.73 7.03 5.68 6.55 5.99 5.60 5.16 0.50 <0.0001 MNT, kgf Thermography a a ab abc bcd cd d bcd cd cd cd de e Max- 36.97 36.97 36.70 36.54 36.12 36.04 35.75 36.07 35.95 35.99 35.93 35.56 34.99 0.25 <0.0001 imum OT, °C a–e Means with like superscripts within each row do not differ (P > 0.05). The concentration of both molecules in plasma de- first 24 h in particular. However, as the study pro- creased over the course of the sampling period. The gressed and pain status likely shifted from acute to change in cortisol concentration over the course of chronic, our sampling regimen should have been this study is typical of studies where blood sam- sufficient to identify differences in chronic sub- pling occurs at multiple time points over several stance P production in response to the disbudding hours to multiple days. It is generally acknowledged lesions. that cortisol concentration typically increases rap- Table 4 shows the impact of sampling time on idly following exposure to acute stressors and ADG, BW, MNT, and   ocular temperature (OT). then declines toward normal concentrations that ADG was not significantly different between sam- vary rhythmically throughout the day, even with pling times (P  =  0.74). However, BW increased the prolonged presence of the same acute stressor across sampling times (P  <  0.0001). Both MNT (Apple et al., 2005). Stock et al. (2015) identified a and OT declined over the sampling period. It is similar time effect on cortisol concentrations in pre- possible that this may have been a result of the weaned dairy calves that were cautery disbudded. adjustment of homeostatic mechanisms in the Our data suggest that the change in substance P neonatal calves following parturition. In a re- and cortisol concentration across all calves over view, Kirovski (2015) described the dependence time may have been resultant of human presence of neonatal calves on nonshivering thermogen- and restraint stress. Investigation of this concept is esis, a process that involves the uncoupling of warranted and necessary. In addition, specific guid- adenosine triphosphate (ATP) synthase from the ance regarding the optimal time of day to measure electron transport chain, resulting in heat produc- substance P is needed for cattle. In our study, we tion in brown adipose tissue. Brown adipose tissue consistently collected all blood for assay during the recedes during the first month of life for bovine same time frame (0500–0700 hours) each day. It is calves and other neonatal ruminants. It is possible possible that the 24-h time lapse between substance that the decline in OT that we observed was an in- P sample collections in our study could have missed dication of the progressive reduction in nonshiver- changes in concentration that occurred during the ing thermogenesis through the first month of life. Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 Oral meloxicam after caustic paste disbudding 9 Table 5. Least squares means for the interaction effect of meloxicam treatment and sampling time on mean horn bud temperature and mean horn bud to ocular temperature ratio of 3-d-old Holstein calves disbudded with caustic paste (N = 61) Sampling time Dependent variable −5 min 0 h 12 h 24 h 48 h 60 h 72 h 96 h 108 h 156 h 204 h 252 h 300 h SE P-value Mean horn bud temperature, °C CONTROL 25.46 36.75 33.55 32.49 32.29 32.86 32.93 33.60 33.66 33.93 34.48 34.02 33.46 0.46 <0.0001 SHAM 25.99 34.57 33.46 33.46 32.13 33.20 32.19 32.94 33.30 33.28 33.40 33.28 33.22 M1 25.19 36.73 33.01 32.76 33.14 33.20 32.79 32.87 33.25 33.60 33.69 33.43 32.93 ab M2 23.96 36.64 33.01 32.14 33.41 33.29 32.60 33.15 33.00 33.45 33.09 33.93 33.21 Mean horn bud: ocular temperature CONTROL 0.68 1.00 0.91 0.89 0.90 0.91 0.92 0.93 0.93 0.94 0.95 0.96 0.96 0.01 <0.0001 SHAM 0.69 0.93 0.91 0.92 0.89 0.92 0.90 0.91 0.92 0.91 0.92 0.93 0.93 M1 0.67 1.00 0.90 0.89 0.91 0.93 0.92 0.91 0.93 0.94 0.95 0.94 0.96 M2 0.65 1.00 0.90 0.88 0.92 0.92 0.92 0.92 0.92 0.94 0.93 0.96 0.95 a,b Means within dependent variable with like superscripts within column do not differ (P > 0.05). Treatments: 1) M1, caustic paste disbudding and oral meloxicam (45 mg) with a placebo 24 h later; 2) M2, treatment M1 followed by a second 45-mg dose of meloxicam 24 h later instead of placebo; 3) CONTROL, treatment M1 with placebo in place of meloxicam; and 4) SHAM, sham disbudding with placebo in place of meloxicam. Glynn et  al. (2013) reported a similar time effect of nondisbudded tissue during a 3-wk period fol- on MNT values proximal to the horns of Holstein lowing disbudding. However, the same project steers. found that calves remained sensitive to touch near Table 5 reports the interaction effects of meloxi- the disbudding site for 9 wk. Overall, our findings cam treatment and sampling time on mean horn align with the conclusions of Coetzee et al. (2012) bud temperature by thermography and mean horn that administration of meloxicam as the only pain bud to ocular temperature ratio. It is important to mitigation therapy for horn tissue removal is insuf- note that the values reported at −5  min for both ficient in the control of acute pain but may be valu- variables occurred before the horn buds were shaved able in the control of chronic pain associated with for paste application. Longer and thicker hair pro- postprocedure healing. It is important to note that vided greater insulation and cooler temperatures at the discussion of the results in this study included the surface of the hair coat that was captured in the reference to an array of studies that focused on the thermal image. Both dependent variables exhibited responses of cattle to horn removal and the im- the treatment by sampling time interaction effect at pact of multiple pain mitigation strategies on those time 0 h, which was immediately after the applica- responses. Due to the limited amount of data re- tion of caustic paste for CONTROL, M1, and M2 garding caustic paste disbudding that was available calves or the noncaustic analog for SHAM. Horn at the time of this publication, reference to other bud temperature was greater in the CONTROL and disbudding methods was necessary. However, the M1 groups than SHAM but not different than M2. specific mechanisms of action are vastly different Mean horn bud to ocular temperature ratio was between cautery, caustic, and mechanical horn re- less in the SHAM group than any treatment that moval and different mechanisms and magnitudes of received caustic paste. No other difference was ob- pain response must be expected and acknowledged. served between treatments at any other sampling Our data suggest that meloxicam, as applied in time. The data for both horn bud temperature and our study, is not sufficient as the primary therapy horn bud to ocular temperature ratio suggest that to control pain and inflammation associated with the heat production associated with apparent tissue caustic paste disbudding. Further study regarding inflammation from caustic paste application did the impact of cornual nerve block in conjunction not last longer than 12 h, regardless of meloxicam with meloxicam administration on caustic paste treatment. It is important to note that the absence disbudded calves is warranted as other previously of heat is not sufficient to conclude that pain has cited work has identified the value of regional anes- subsided. Adcock and Tucker (2018) found that thesia to controlling the acute pain associated with the surface temperature of cautery disbudding caustic paste disbudding and the need for effective wounds did not differ from the surface temperature chronic pain mitigation. The variables that we Translate basic science to industry innovation Downloaded from https://academic.oup.com/tas/advance-article-abstract/doi/10.1093/tas/txz151/5610601 by DeepDyve user on 03 December 2019 10 Karlen et al. Coetzee,  J.  F., R.  A.  Mosher, B.  KuKanich, R.  Gehring, utilized in this study may not have been completely B.  Robert, J.  B.  Reinbold, and B.  J.  White. 2012. effective in quantifying the magnitude of chronic Pharmacokinetics and effect of intravenous meloxi- pain associated with caustic paste administra- cam in weaned Holstein calves following scoop dehorn- tion as no overall effect was observed between the ing without local anesthesia. BMC Vet. Res. 8:153. CONTROL, M1, and M2 treatment groups over doi:10.1186/1746-6148-8-153 Cotter,  F . 2013. A GLP tissue residue depletion study in calves fol- time. This study would have benefitted from inclu- lowing oral / buccal administration of meloxicam. Available sion of assays of behavioral indicators of pain and from https://www.mla.com.au/research-and-development/ discomfort. Preliminary work in our lab has sug- search-rd-reports/final-report-details/Animal-Welfare/A- gested that behaviors associated with chronic pain GLP-tissue-residue-depletion-in-calves-following-oral- and discomfort may be subtle in young calves and buccal-administration-of-meloxicam/2072#. Accessed considerable care must be taken to quantify behav- May 31, 2019. European Medicines Agency (EMA). 2018. Metacam (meloxi- iors that effectively represent changes in pain status. cam): An overview of Metacam and why it is authorized in A behavioral assay with capability to detect pain-in- the EU. Available from https://www.ema.europa.eu/docu- duced pessimism and anhedonia has recently been ments/overview/metacam-epar-summary-public_en.pdf. suggested by other researchers for the assessment Accessed February 25, 2019. of mood changes in calves that were cautery dis- Fraccaro, E., J. F. Coetzee, R. Odore, L. N. Edwards-Callaway, B.  Kukanich, P.  Badino, L.  Bertolotti, H.  Glynn, budded (Lecorps et al., 2019). We recommend that J. Dockweiler, K. Allen, et al. 2013. A study to compare future work to investigate the impact of pain miti- circulating flunixin, meloxicam and gabapentin concen- gation strategies on pain response in calves, specif- trations with prostaglandin E₂ levels in calves under- ically, and animals, in general, include variables that going dehorning. Res. Vet. Sci. 95:204–211. doi:10.1016/j. effectively quantify both physiological and behav- rvsc.2013.01.018 ioral indicators of changes to affective states. 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Published: Nov 1, 2019

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