Effects of a Multimodal Pain Control Protocol Using 2 Lidocaine Intradermal and Meloxicam Intramuscular on Mitigating Behavioral Castration Pain in P

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Journal of Applied Animal Welfare Science ISSN: 1088-8705 (Print) 1532-7604 (Online) Journal homepage: www.tandfonline.com/journals/haaw20 Effects of a Multimodal Pain Control ProtocolUsing 2% Lidocaine Intradermal and MeloxicamIntramuscular on Mitigating BehavioralCastration Pain in Piglets Using a NeedlelessSystemErin Elizabeth King-Podzaline, Gabriella-Louise Stephen, Alexandria Bokhart,Pedro Henrique Esteves Trindade, Victoria Rocha Merenda & MoniqueDanielle Pairis-GarciaTo cite this article: Erin Elizabeth King-Podzaline, Gabriella-Louise Stephen, AlexandriaBokhart, Pedro Henrique Esteves Trindade, Victoria Rocha Merenda & Monique DaniellePairis-Garcia (2026) Effects of a Multimodal Pain Control Protocol Using 2% LidocaineIntradermal and Meloxicam Intramuscular on Mitigating Behavioral Castration Pain in PigletsUsing a Needleless System, Journal of Applied Animal Welfare Science, 29:2, 215-226, DOI:10.1080/10888705.2024.2440894 To link to this article: https://doi.org/10.1080/10888705.2024.2440894 © 2024 The Author(s). Published by InformaUK Limited, trading as Taylor & FrancisGroup. View supplementary material Published online: 18 Dec 2024. Submit your article to this journal Article views: 659 View related articles View Crossmark data Full Terms & Conditions of access and use can be found athttps://www.tandfonline.com/action/journalInformation?journalCode=haaw20

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RESEARCH ARTICLE Effects of a Multimodal Pain Control Protocol Using 2% Lidocaine Intradermal and Meloxicam Intramuscular on Mitigating Behavioral Castration Pain in Piglets Using a Needleless System Erin Elizabeth King-Podzaline a , Gabriella-Louise Stephen a , Alexandria Bokhart a , Pedro Henrique Esteves Trindade b , Victoria Rocha Merenda b , and Monique Danielle Pairis- Garcia b a Department of Ambulatory Medicine and Theriogenology, Tufts University Cummings School of Veterinary Medicine, North Grafton, MA, USA; b College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA ABSTRACT Mitigating piglet castration pain is critical to minimize negative experiences and contribute to a positive affective state. A multimodal pain control protocol (needleless 2% lidocaine intradermal/0.4 mg/kg meloxicam intra - muscular) was evaluated. Males were administered one of four treatments: (1) needleless lidocaine/meloxicam intramuscular (LM), (2) needleless lido - caine/saline intramuscular (LS), (3) needleless saline/meloxicam intramuscu - lar (SM), and (4) needleless saline/saline intramuscular (SS). Females were sham castrated (SH). Piglets were recorded 24h pre-castration (M1) and 15min (M2), 3h (M3), and 24h post-castration (M4). The Unesp-Botucatu Composite Acute Pain Scale (UPAPS) was used to assess pain behavior. From a treatment standpoint, scores did not differ at M1 ( P = 1.00) or M4 ( P ≥ 0.36). However, at M2, LS piglets had ( P < 0.01) higher scores (3.4) than LM piglets (1.6) and SH piglets had ( P ≤ 0.01) the lowest scores (0.02). From a timepoint standpoint, piglets in the LS, LM, SM, and SS groups had ( P ≤ 0.05) higher scores at M2 compared with their baseline scores at M1. These results indicate that needleless lidocaine and

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SM, and SS groups had ( P ≤ 0.05) higher scores at M2 compared with their baseline scores at M1. These results indicate that needleless lidocaine and intramuscular meloxicam offered no analgesic benefit. ARTICLE HISTORY Received 18 June 2024 Accepted 26 November 2024 KEYWORDS Swine castration; pain scale; animal welfare; lidocaine; needleless Introduction There are an estimated 94 million piglets surgically castrated in the United States per year (Wagner et al., 2020 ). Surgical castration is a critical animal welfare issue because it is a painful procedure that results in both acute and chronic pain sensitivity (Hay et al., 2003 ; Ison et al., 2016 ; Kluivers-Poodt et al., 2013 ). When assessing welfare from the Five Domains Model (Mellor & Beausoleil, 2015 ; Mellor et al., 2020 ), castration negatively impacts the fifth domain (i.e., mental state) as demon - strated by changes to the animal’s behavioral repertoire and physiological homeostasis (Moya et al., 2008 ; Yun et al., 2019 ). Post-castration pain and stress alters piglets’ blood physiology profiles, including but not limited to pH, lactate and hematocrit (Pérez-Pedraza et al., 2018 ). Previous research has found that post-castration pain contributes to many behavioral changes including, but not limited to, posture (i.e., changing posture with discomfort, tense and/or back arched), activity (i.e., decreased movement, restlessness, or reluctance to move) and interaction with the environment (i.e., decreased interaction with littermates and/or disinterested in surroundings) (Luna CONTACT Erin Elizabeth King-Podzaline erin.kingdvm@tufts.edu Tufts University Cummings School of Veterinary Medicine, 200 Westboro Rd, North Grafton, MA 01536, USA Supplemental data for this article can be accessed online at

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ity Cummings School of Veterinary Medicine, 200 Westboro Rd, North Grafton, MA 01536, USA Supplemental data for this article can be accessed online at https://doi.org/10.1080/10888705.2024.2440894 . © 2024 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License ( http:// creativecommons.org/licenses/by-nc-nd/4.0/ ), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent. JOURNAL OF APPLIED ANIMAL WELFARE SCIENCE 2026, VOL. 29, NO. 2, 215–226 https://doi.org/10.1080/10888705.2024.2440894

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et al., 2020 ). Although this procedure is painful, analgesia and local anesthesia at the time of castration is not commonly administered in the US (Wagner et al., 2020 ). Pain management on US swine farms is limited primarily due to the lack of Food and Drug Administration (FDA) approved products labeled for pain control in swine (Pérez-Pedraza et al., 2018 ). Therefore, identifying realistic and effective pharmaceutical products that can be approved for use on farms is needed in the US. Lack of pain management at the time of castration may also be due to the idea that neonates due not perceive pain as severely due to underdeveloped nervous systems at birth, despite empirical evidence otherwise (Adcock, 2021 ). In order to mitigate piglet castration acute and chronic pain effectively, a multimodal analgesic approach must be considered and take into account logistical and economic considerations for the producer (Wagner et al., 2020 ). Currently, non-steroidal anti-inflammatories (NSAIDs) and local anesthetics are promising products for pain management for large commercial swine systems that can be used in association to control operative and post-operative pain. Meloxicam, a COX-2 specific NSAID, is the most frequently studied NSAID used for castration pain mitigation in piglets (Bates et al., 2014 ; Baysinger et al., 2021 ; Bonastre et al., 2016 ; Burkemper et al., 2020 ; Enouri et al., 2022 ; Garcia et al., 2023 ; Gottardo et al., 2016 ; Hansson et al., 2011 ; Keita et al., 2010 ; Nixon et al., 2021 ; Viscardi & Turner, 2018 ) and controls prostaglandin production and pain sensitivity up to 5 hours post-procedure (Engelhardt, 1996 ; Gottardo et al., 2016 ). Lidocaine is a local anesthetic that blocks voltage-gated sodium channels thus preventing an effective action

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elhardt, 1996 ; Gottardo et al., 2016 ). Lidocaine is a local anesthetic that blocks voltage-gated sodium channels thus preventing an effective action potential from occurring (Hermanns et al., 2019 ). Local anesthetics like lidocaine are required to be used in many European countries for pain control during castration and function to control pain at the time of the procedure (European Commission, 2017 ). Although meloxicam administration can be easily implemented on-farm, effective and consistent administration of local anesthetics requires more extensive training (European Commission, 2017 ; Pérez-Pedraza et al., 2018 ). More recently, the implementation of a needleless administration system has been explored, and to date, only one publication has assessed the efficacy of a needle-free injection of lidocaine to control castration pain in piglets (M. Sutherland et al., 2017 ). Needle-free systems offer many advantages in terms of food safety, worker safety and animal welfare. Needle-free systems are relatively cost-effective depending on size of the farm and farm needs and this is mostly due to a reduction in labor savings with needleless systems being quicker and safer to manage (Imeah et al., 2020 ). These systems eliminate needle fragments in carcasses (Hoff & Sundberg, 1999 ), as well as reduce the risk of worker injuries from handling used needles (Chase et al., 2008 ). Needle-free systems used for lidocaine administration have been proven to be less painful and cause less stress in humans (Cooper et al., 2000 ) and swine (M. Sutherland et al., 2017 ) over traditional injections. With needle-free systems, tissue trauma from a conventional needle is eliminated. The product is dis - tributed evenly through micro skin openings in the dermis through either

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tissue trauma from a conventional needle is eliminated. The product is dis - tributed evenly through micro skin openings in the dermis through either spring-power, battery- power or compressed-gas power, which leads to instant dispersion and better absorption (Temple et al., 2020 ). Therefore, the aim of this study was to determine the effect of a multimodal pain control protocol (0.4 mg/kg meloxicam IM and 2% lidocaine intradermal) on mitigating behavioral castration pain using a novel needleless system. Materials and methods Study animals/housing This study was approved under Tufts University IACUC Protocol #G2021–140. Male and female purebred Yorkshire piglets between three and 12 days of age were enrolled. A total of 96 males underwent the surgical castration procedure and 78 females were sham castrated from 18 litters. Only healthy animals were enrolled. Small piglets, cryptorchids, or lame piglets were not included, representing a total of three piglets across the 18 litters. Piglets were tail docked with a hot iron and ear notched as part of routine husbandry care between day one and three of life. Research suggests 216 E. E. KING-PODZALINE ET AL.

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that pain associated with hot iron docking subsides within 28 hours post-procedure (Morrison & Hemsworth, 2020 ). Therefore, for this study, castration was performed at a minimum of 48 hours post-docking and ear notching. Sows and piglets were housed in individual farrowing crates on fully slatted floors at Tufts University Cummings School of Veterinary Medicine (CSVM) Swine Unit II in North Grafton, Massachusetts. The farrowing room was mechanically ventilated, and the temperature was main - tained at 18°C. Light was provided daily from 600-1800 h. Creep area for piglets was heated to approximately 32–35°C using heating lamps. Sows had ad libitum access to one water nipple and feeder (diet met or exceeded National Research Council nutrient requirements for lactating sows) (Council National Research, 2012 ). Treatments Piglets were weighed, and males were randomly assigned to one of four surgical castrated treatments (LM, LS, SM, or SS), while female piglets were included in a single sham castrated treatment (SH) described below. Piglets were assigned a number in consecutive order and were marked. Treatment groups were evenly assigned to the 96 males using a random number generator and this information was kept in an Excel file. Those administering treatments, performing castrations, and assessing behavioral response were blind to treatment. ● LM : 8 mg/kg lidocaine (VetOne, 2%) administered locally over scrotum via a needleless injector (Pulse 50 Lenexa, Kansas) + 0.4 mg/kg meloxicam (VetOne, 5 mg/ml) intramuscularly prior to castration ● LS : 8 mg/kg lidocaine administered locally over scrotum via a needleless injector +0.2 ml saline (VetOne, 0.9%) intramuscularly prior to castration ● SM : 0.5 ml sterile saline administered locally over scrotum via a

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leless injector +0.2 ml saline (VetOne, 0.9%) intramuscularly prior to castration ● SM : 0.5 ml sterile saline administered locally over scrotum via a needleless injector +0.4 mg/kg meloxicam intramuscularly prior to castration ● SS : 0.5 ml sterile saline administered locally over scrotum via a needleless injector +0.2 ml saline intramuscularly prior to castration ● SH : Female control group received no drug administration prior to sham castration Surgical castration procedure Five, second-year veterinary students from CSVM performed all castration and treatment adminis - tration on enrolled male piglets ( n = 96) and sham castrations on female piglets ( n = 78) weekly on the same day, at the same time between 1300-1500 h over a 13-week period in the spring of 2021. Students were first trained on the procedure, both in a clinical skills laboratory as part of their curriculum and in an additional 2-hour training session prior to this study. All castrations were supervised by the veterinarian. Castrations were performed equitably between all five veterinary students. Piglets were removed from their farrowing crate via a transport cart to a nearby room for castration. Piglets were weighed and administered assigned treatments 10–15 minutes prior to castration procedure. For administration of treatments via the needleless injector, piglets were placed in a v-trough, the correct dose was set and the injector was held flat against the entirety of the scrotum while the device was activated to inject either lidocaine or saline intradermally into the scrotum. For the castration procedure, piglets were placed in a v-trough and a scrotal castration was performed through one horizontal incision using a #10 blade, as described by Viscardi et al. ( 2017 ). One testicle was

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h and a scrotal castration was performed through one horizontal incision using a #10 blade, as described by Viscardi et al. ( 2017 ). One testicle was retracted and pulled away from the body and subsequent cutting of the spermatic cord was then performed. The same was repeated for the other testicle. The wound was sprayed with JOURNAL OF APPLIED ANIMAL WELFARE SCIENCE 217

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iodine. Once the entire litter was done, the piglets were immediately returned to their farrowing crate. Video recording behavior Piglets were recorded using high-definition video cameras (Kicteck full HD 1080P, 15 FPS, 24 MP) mounted 1.2 m above the ground and angled at 45° so that the entire crate was visible. Behavior for each piglet was recorded continuously for 1 h in duration for the following four recording periods: M1 (24 h pre-procedure), M2 (15 min post-procedure), M3 (3 h post-procedure), M4 (24 h post- procedure). At the end of the trial, veterinary students not involved in the behavioral analysis downloaded all videos. One, four-minute video clip was trimmed from each recording timepoint (M1-M4). Video clips were obtained at the 9-min mark for each recording period (i.e., 9 min after the start of each recording period) until the 12-min mark. The 9-min mark was randomly selected to eliminate observer bias. If all piglets were sleeping and/or nursing from the 9–12 min mark, the 2–6 min mark was selected instead. Once the continuous 4-min video clips were downloaded, clips were rando - mized and masked for identifying information (i.e., time stamp, date etc.) prior to initiation of pain scale validation (72 total 4-min video clips). Piglets were excluded from video analysis if their entire body was not visible for the duration of the video. Observer training and behavioral assessment The video clips from this study were analyzed by three female observers. Prior to data analysis, the observers underwent 5 training sessions (2 h/training session). During training sessions, video links provided in the UPAPS ( Table 1 ) (Luna et al., 2020 ) were viewed and discussed in a group. Additional training sessions included assessing practice videos both within the

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( Table 1 ) (Luna et al., 2020 ) were viewed and discussed in a group. Additional training sessions included assessing practice videos both within the group and individu - ally. Upon completion of the training, each observer evaluated and scored 10 test videos provided on animalpain.org. Observers were considered ready to use the scale when their scores were up to 20% higher or lower than template scores. Bias could exist if observers could differentiate piglet gender for behavioral observation. Observers assessed no more than two piglets at a time for each 4-minute timepoint. Statistical analysis All data were analyzed using SAS (version 9.4; SAS Institute Inc). A multivariable model was built at the piglet level for the total behavioral scores using the UPAPS scale (outcome of interest). The total behavioral scores were analyzed as repeated measures by an analysis of variance applying a multilevel model, and the structure of the model was chosen according to the smallest Bayesian information criterion. Treatment, timepoint, and the interaction between treatment and timepoint were included as fixed effects. The P-values for all pairwise comparisons among all treatments were adjusted using the Tukey-Kramer method. Statistical significance was declared at p ≤ 0.05. Results Study population All piglets were observed and scored for all timepoints, however, one piglet was diagnosed with severe lameness. Due to the potential confounding effects of pain associated with this condition, this piglet was excluded from the study and analysis. To ensure the accuracy of the pain scale, piglets were only observed during timepoints if they were awake. Across M1-M4, 401 out of 537 timepoints were included with the remaining timepoints ( n = 136) excluded from the analysis because

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if they were awake. Across M1-M4, 401 out of 537 timepoints were included with the remaining timepoints ( n = 136) excluded from the analysis because the piglet 218 E. E. KING-PODZALINE ET AL.

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Table 1. The Unesp-Botucatu pig composite acute pain scale (Luna et al., 2020 ). Item Score Score/criterion Links to videos Posture 0 Normal 1 (any position, apparent comfort, relaxed muscles) or sleeping https://youtu.be/ ojvbFAgCWo0 1 Changes posture, with discomfort 2 https://youtu.be/ SpaWsFCrPxE 2 Changes posture, with discomfort, and protects the affected area 3 https://youtu.be/ VjSlsRrG8yA 3 Quiet, tense, and back arched 4 https://youtu.be/ gHUWWf5836M Interaction and interest in the surroundings 0 Interacts with other animals; interested in the surroundings 5 or sleeping https://youtu.be/ 6s8xEn3QR3c 1 Only interacts if stimulated by other animals; interested in the surroundings. https://youtu.be/ 9pMULbCO_WE 2 Occasionally moves away from the other animals, but accepts approaches 6 ; shows little interest in the surroundings https://youtu.be/ hp4wqdcHanA 3 Moves or runs away from other animals and does not allow approaches; disinterested in the surroundings https://youtu.be/ se70oYXcWFw Activity 0 Moves normally 7 or sleeping https://youtu.be/ cC75t7L5-YA 1 Moves with less frequency 8 https://youtu.be/ lQo9wq8Lan8 2 Moves constantly, restless 9 https://youtu.be/ YQRJjijLvpk 3 Reluctant to move or does not move 10 https://youtu.be/ Zyx0G3Wpt8o Nursing* 0 Actively nursing https://youtu.be/ 6e2iYNKVFNY 1 Not nursing when less than 50% of the piglets are nursing https://youtu.be/ 3xJuMo2t7b4 2 Unsuccessful attempt at nursing (one or more attempts) https://youtu.be/ 3PLgcf9ceQ8 3 Not nursing when 50% or more of the piglets are nursing https://youtu.be/ TZcB5PbsTT8 Attention to the affected area A. Elevates pelvic limb or alternates the support of the pelvic limb https://youtu.be/ UD99ftO7HE0 B. Scratches or rubs the painful area 11 https://youtu.be/ 7idfFk1harE C.

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b or alternates the support of the pelvic limb https://youtu.be/ UD99ftO7HE0 B. Scratches or rubs the painful area 11 https://youtu.be/ 7idfFk1harE C. Moves and/or runs away and/or jumps after injury of the affected area https://youtu.be/ u-Pqubom278 D. Sits with difficulty https://youtu.be/ ETNEOCVV4h0 0 All the above behaviors are absent 1 Presence of one of the above behaviors 2 Presence of two of the above behaviors 3 Presence of three or all the above behaviors Miscellaneous behaviors A. Wags tail continuously and intensely 12 https://youtu.be/ pU5dGZFNRHc B. Bites the bars or objects https://youtu.be/ cF3dsq7gMtk C. The head is below the line of the spinal column. https://youtu.be/ ZcIgngclRpI D. Presents difficulty in overcoming obstacles (example: other animal) https://youtu.be/ HlvdOI3lGuY 0 All the above behaviors are absent 1 Presence of one of the above behaviors 2 Presence of two of the above behaviors 3 Presence of three or all the above behaviors 1 Defined as any position in which the piglet demonstrates apparent comfort and relaxed muscles. Postures considered normal which demonstrate piglet comfort include but are not limited to piglets laying in lateral or sternal recumbency either separately or amongst the litter. Normal postural changes include a quick transition from standing to lying and lying to standing and typically result in the entire body descending onto or away from the floor at relatively the same time. Postural positions are most likely driven by external stimuli (e.g., littermate stands up or over piglets while lying). While standing, the piglet’s head is parallel to the spine and the pig will frequently explore the environment by changing head positions (elevating head to explore surrounding area and placing head toward floor to

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he pig will frequently explore the environment by changing head positions (elevating head to explore surrounding area and placing head toward floor to explore ground). JOURNAL OF APPLIED ANIMAL WELFARE SCIENCE 219

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was sleeping. Four additional timepoints were excluded as these piglets who were not visible for the entirety of the video. UPAPS scores between treatments and over time The adjusted P -values for pairwise comparisons between treatments over timepoints and within treatments over timepoints are depicted in supplementary table S1 and S2, respectively. There was a treatment by timepoint ( p < 0.0001), treatment ( p < 0.0001) and timepoint effect ( p < 0.0001) on total behavioral scores using the UPAPS scale ( Figure 1 ). The total behavioral scores did not differ between treatments at M1 ( p = 1.00) or M4 ( p ≥ 0.36). However, at M2, piglets in the LS group had ( p < 0.01) higher scores (3.4) than piglets in the LM group (1.6) and piglets in the SH group had ( p ≤ 0.01) lower scores (0.02) than piglets in the LM (1.6), LS (3.4), SM (2.2), SS (2.9) groups. Piglets in the LM, LS, SM and SS groups had ( p ≤ 0.05) higher scores at M2 compared with their respective baseline at M1. Piglets in the LS group had ( p ≤ 0.0001) higher scores (3.4) at M2 than at M3 (1.0) and at M4 (1.1). Similarly, piglets in the SS group had ( p = 0.004) higher scores M2 (2.86) compared with its scores at M3 (0.25). The scores of SH piglets did not change at any timepoint ( p = 1.00). 2 Defined as a piglet demonstrating a deviation from normal behavior in which muscles may be tensed, postural change transitions occur more slowly and may occur more frequently. Piglets demonstrating changing posture with discomfort may often extend the rear legs to elevate the rump off the ground while maintaining physical contact to the floor with the front legs and head. Transition from standing to lying or lying to standing also occurs more slowly in which the piglet may remain motionless for short time

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legs and head. Transition from standing to lying or lying to standing also occurs more slowly in which the piglet may remain motionless for short time periods and change posture in the absence of external stimuli such as other littermates. 3 Demonstrated behaviorally as a piglet performing a tucked tail posture in which the rear legs are slight flexed, and the piglet orients the castration site toward the walls or corner of the farrowing stall or runs away when another pig touches the affected area. 4 Typically accompanied with little to no movement or activity. Back has a defined arch, head of the piglet is below the level of the spine and piglet spends majority of the time not moving or within a small physical space of the pen. 5 Animal interaction includes snout to snout contact with littermates but may also include rooting or nosing body parts of littermates including the flank, neck, and rear of the littermate of interest. Interest in the surrounding environment includes actively exploring area (i.e., walking, running to various parts of the stall) and actively rooting with the snout or mouthing items within the stall including but not limited to heat mat, bars, and floor. 6 Accepts approach is to reciprocate interaction with another littermate without running away from littermate. 7 Defined as exhibiting forward motion with all four legs in contact with the floor typically at a walking pace. Normal movement examples for piglets include, but is not limited to, exploring the stall floor by walking to and from areas surrounding the sow (with and without the presence of other littermates), attempting to nurse sow by rooting at udder, and interacting with littermates on heat mat. 8 Defined as the piglet still showing activity but is less likely to move for long

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by rooting at udder, and interacting with littermates on heat mat. 8 Defined as the piglet still showing activity but is less likely to move for long periods or cover larger areas of the stall and may not attempt to explore a part of the stall in which no other littermates are present, may not participate in social interactions with littermates, or their behavior deviates from that of littermates. 9 Piglets will likely demonstrate activity at a fast walk or running pace for the majority of the observation. Restless piglets may also demonstrate startle behaviors in which the piglet will quickly freeze (standing with head elevated but not moving) and then resume fast-paced movement. 10 Piglets are likely to be in a lying position for the majority of the observation and demonstrate behavior similar to moribund animals. Piglets reluctant to move will not be easily stimulated by littermates and may isolate themselves from littermates and reside in the corner of the stall. Some piglets may show this behavior while demonstrating a posture score of 3 and will show very little to no reactivity of activity occurring in the pen. Postural changes may occur but will be less frequent in nature and will likely result in little to no physical distance changed from original posture location. 11 Requires physical contact of the surgical site with the farrowing environment (i.e., floor, penning, mat) or littermate. Piglets often perform scratching behavior in which the rear leg is extended to the ear, but this is not considered for purposes of scoring this item as it does not involve the painful area. 12 Tail wagging with intensity is described both by the force in which the tail moves and the frequency. Intense tail wagging will involve movement of the entire tail and movement will

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described both by the force in which the tail moves and the frequency. Intense tail wagging will involve movement of the entire tail and movement will occur in short rapid bursts (3+ more tail wags consecutively). Tail wagging can occur while the piglet is standing, lying, or sitting. *Nursing behavior was observed and analyzed in the study but was excluded from the final pain scale table as it did not meet any of the validation criteria (principal component analysis, specificity, internal consistency, item-total correlation and responsiveness). 220 E. E. KING-PODZALINE ET AL.

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Discussion There continues to be a need to find both practical and effective ways of mitigating pain at the time of castration, as it is known to be a painful procedure with welfare implications for piglets (Wagner et al., 2020 ). The objective of this study was to determine the effect of a multimodal pain control protocol including meloxicam administered intramuscularly and lidocaine administered intrader - mally on mitigating pain behaviors associated with castration. In the present study, piglets that underwent castration displayed greater total pain scores over time compared to sham castrated piglets. These results agree with decades of research that show that castration is painful (Lou et al., 2022 ; Schmid & Steinhoff-Wagner, 2022 ; M. A. Sutherland, 2015 ) and including a male (Garcia et al., 2023 ; Lopez-Soriano et al., 2022 , 2023 ) or female (Robles et al., 2023 ) sham treatment group in castration studies help establish an objective baseline model for comparison. The Unesp-Botucatu Pig Composite Acute Pain Scale (UPAPS) continues to be a robust tool for pain assessment that demonstrates good intra- and inter-observer agreement, excellent predictive capabilities and responsiveness in painful and pain-free states (Robles et al., 2023 ). Although painful (M2-M4) and pain-free (M1) timepoints were different in this study, average pain scores at M1 were lower in this study (0–0.06) compared to previous work conducted by Lopez- Soriano et. al. (0.7–1.3) (Lopez-Soriano et al., 2023 ). Multiple factors may have influenced these differences including but not limited to gender and/or experience of the observers. Work conducted in 2020 by Navarro and colleagues (Navarro et al., 2020 ) demonstrated a clear observer gender effect with women observers assessing higher

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conducted in 2020 by Navarro and colleagues (Navarro et al., 2020 ) demonstrated a clear observer gender effect with women observers assessing higher pain scores than male observers. This was not the case in our study and further work is needed to evaluate how factors such as gender may impact pain assessment both within pigs and across species. Lidocaine (LS treatment) had no effect on mitigating total pain scores in piglets immediately post- castration. This is in agreement with previous work showing no effect of injectable lidocaine on decreasing pain-associated behaviors for piglets undergoing castration (Kluivers-Poodt et al., 2013 ; Figure 1. Effect of treatment on total behavioral scores using UPAPS scale in castrated piglets treated with needleless lidocaine and meloxicam intramuscular (LM), needleless lidocaine and saline intramuscular (LS), needleless saline and meloxicam intramus - cular (SM), needleless saline and saline intramuscular (SS) or sham castrated females (SH). Treatment p < .0001, timepoint p < .0001, treatment x timepoint p = <.0001. Pairwise comparisons between treatments with p ≤0.05 at 15 min: LM vs. LS, LM vs. SH, LS vs. SH, SM vs. SH, SS vs. SH. JOURNAL OF APPLIED ANIMAL WELFARE SCIENCE 221

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Lopez-Soriano et al., 2023 ; Zankl et al., 2007 ). However, lidocaine-treated piglets demonstrated increased pain scores at 3 h post-castration compared to all other treatment groups. While generally considered less painful than a conventional needle, needleless administration of lidocaine could have resulted in pain experienced by the piglet in conjunction to the castration procedure. The needleless device used in this study had a similar diameter to the scrotum of the piglet. It could be inferred that such a highly pressurized delivery of lidocaine over a much greater surface area than a conventional needle led to some level of pain experienced by the piglet. In humans, pain has been reported with lidocaine needleless injection when used prior to IV catheter insertion (Stoltz & Manworren, 2017 ) and periodontal procedures (Gupta et al., 2018 ), as compared to other anesthetic delivery methods. Additionally, the lidocaine was not buffered with an alkalinizing agent and could have led to an increased pain response, as has been shown in humans (Brandis, 2011 ; Parham & Pasieka, 1996 ). Interestingly, LM piglets did not demonstrate increased scores at 3 h. This may be due to the analgesic benefits of meloxicam for injection site pain. In this study, we targeted lidocaine to be administered intradermally into the scrotum of the piglets and not intratesticular. Previous studies have demonstrated that lidocaine injected using a needle is effective in mitigating castration pain when the drug is administered intratesticular and scrotal (Haga & Ranheim, 2005 ; Werner et al., 2022 ). Failure to mitigate pain in this study with lidocaine may be due to the fact that the administration location was not consistent and did not permit lidocaine to be absorbed proximally into the

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h lidocaine may be due to the fact that the administration location was not consistent and did not permit lidocaine to be absorbed proximally into the spermatic cord providing no significant anesthetic effects at the surgical site. Reevaluating the location for lidocaine injection is needed for future work with the needleless device. Similar to LS treatment results, meloxicam (SM) administered prior to castration had no benefit in pain mitigation post-castration. This agrees with previous work finding no benefit of meloxicam reducing behavioral or physiological indicators of pain associated with castration either immediately or 24 h post-castration (Garcia et al., 2023 ). However, results from the current study are in contrast with existing literature on the benefits of meloxicam for piglets post-castration, with some work demonstrating a reduction in pain behavior and/or serum cortisol for those administered meloxicam compared to castrated controls (Gottardo et al., 2016 ; Langhoff et al., 2009 ; Nixon et al., 2021 ; Schmidt et al., 2012 ). This is likely associated with the variability of the behavioral methodologies used in these studies. To the author’s knowledge, no studies to date have evaluated meloxicam efficacy for mitigating castration pain in pigs using a pain scale. Therefore, a direct comparison between these studies may be inappropriate. It should be expected that given the lack of pain mitigation with lidocaine and meloxicam administered individually, there was no benefit for pain relief when lidocaine and meloxicam were administered concurrently (LM). This is in contrast with other studies that have demonstrated a reduction in serum cortisol and pain behavior when lidocaine and meloxicam were administered together (Bonastre et al., 2016 ). Limitations

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nstrated a reduction in serum cortisol and pain behavior when lidocaine and meloxicam were administered together (Bonastre et al., 2016 ). Limitations previously discussed in this paper specific to adminis - tration technique and behavioral evaluation are likely at play with these results and further research is needed to determine the true benefit of this multimodal approach for pain relief. Total pain scores over time reduced significantly from 15 min post-castration to 3 h post- castration. This is consistent with others (Lopez-Soriano et al., 2023 ) who found no difference in total pain scores at 3 h post-castration compared to baseline. This likely indicates that the acute pain response occurs prior to 3 h post-castration. Another factor contributing to this reduction in total pain scores could be an overall decrease in painful behavior displayed by the piglet due to an increase in nursing frequency, which was not measured in this study. The literature is inconsistent regarding the effects of castration on nursing behavior. While some studies have found a decrease in the amount of suckling time (Mcglone & Hellman, 1988 ; Moya et al., 2008 ), others have found that piglets spent more time massaging the udder post-castration (McGlone et al., 1993 ). Others, however, found that castration does not change nursing behavior (Caroll et al., 2006 ; Rault & Lay, 2011 ; Taylor et al., 2001 ). The current study is not free of limitations. Although there was a possibility for observers to recognize female piglets (SH), all videos were evaluated in a random order to minimize this potential 222 E. E. KING-PODZALINE ET AL.

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bias. Including a female sham treatment group in castration studies helps to establish an objective baseline model for comparison (Robles et al., 2023 ), but future studies should consider using male piglets for sham castration treatment to minimize any potential for a sex effect on behavioral outcomes. In addition, the piglets in this study were subjected to two painful procedures (ear notching and tail docking) prior to castration as part of the farm’s routine management. Piglets were castrated after a period of 48 hours in which signs of pain-induced tail docking or ear notching were no longer observed, minimizing interference in the current study. This is in agreement with existing literature that pain associated with hot iron docking subsides within 28 hours post- procedure (Morrison & Hemsworth, 2020 ). Furthermore, pain behavior is understood to be complex in prey species and this could have contributed to differences between the present study and other studies assessing the efficacy of lidocaine and meloxicam for castration pain. Future work could also include other physiological measures alongside total pain scores. The needleless system is a novel technique that appears promising from both an animal welfare perspective and for worker safety and ease of administration. Future studies should integrate a more comprehensive approach to quantifying pain including the evaluation of other physiological bio - markers. Future work within the swine industry should continue to focus on finding specific pharmaceuticals and analgesic protocols to mitigate post-castration pain in piglets given the sig - nificant animal welfare implications associated with the procedure. Acknowledgments Research was supported with discretionary funds allocated by the Department of

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mal welfare implications associated with the procedure. Acknowledgments Research was supported with discretionary funds allocated by the Department of Ambulatory Medicine and Theriogenology at Tufts University Cummings School of Veterinary Medicine. The authors gratefully acknowledge the farm staff at Tufts University Cummings School of Veterinary Medicine for their logistical support. Disclosure statement No potential conflict of interest was reported by the author(s). Funding The work was supported by the Department of Ambulatory Medicine and Theriogenology at Tufts University Cummings School of Veterinary Medicine. References Adcock, S. J. J. ( 2021 ). Early life painful procedures: Long-term consequences and implications for farm animal welfare. Frontiers in Animal Science , 2 . https://doi.org/10.3389/fanim.2021.759522 Bates, J. L., Karriker, L. A., Stock, M. L., Pertzborn, K. M., Baldwin, L. G., Wulf, L. W., Lee, C. J., Wang, C., Coetzee, J. F., & Staffieri, F. ( 2014 ). Impact of transmammary-delivered meloxicam on biomarkers of pain and distress in piglets after castration and tail docking. PLOS ONE , 9 (12), e113678. https://doi.org/10.1371/journal. pone.0113678 Baysinger, A., Webb, S. R., Brown, J., Coetzee, J. F., Crawford, S., DeDecker, A., Karriker, L. A., Pairis-Garcia, M., Sutherland, M. A., & Viscardi, A. V. ( 2021 ). Proposed multidimensional pain outcome methodology to demonstrate analgesic drug efficacy and facilitate future drug approval for piglet castration. Animal Health Research Reviews , 22 (2), 163–176. https://doi.org/10.1017/S1466252321000141 Bonastre, C., Mitjana, O., Tejedor, M., Calavia, M., Yuste, A., Úbeda, J., & Falceto, M. ( 2016 ). Acute physiological responses to castration-related pain in piglets: The effect of two local

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