Source: UNIVERSITY OF ARKANSAS submitted to NRP
MANAGING PAIN IN CATTLE UNDERGOING CASTRATION USING A BIODEGRADABLE MICRONEEDLE PATCH CONTAINING MELOXICAM
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
ACTIVE
Funding Source
Reporting Frequency
Annual
Accession No.
1027329
Grant No.
2021-67015-35675
Cumulative Award Amt.
$500,000.00
Proposal No.
2021-06299
Multistate No.
(N/A)
Project Start Date
Aug 1, 2021
Project End Date
Jul 31, 2025
Grant Year
2021
Program Code
[A1251]- Animal Health and Production and Animal Products: Animal Well-Being
Recipient Organization
UNIVERSITY OF ARKANSAS
(N/A)
FAYETTEVILLE,AR 72703
Performing Department
(N/A)
Non Technical Summary
Microneedle patches for transdermal drug delivery have shown promise in human healthcare but have been underutilized in veterinary medicine. Current approaches for administering medication to livestock suffer from harsh application approaches. We propose that a patch-type drug delivery method could simplify the industry's need for longer term drug administration. Our research team will plan to developed a biodegradable microneedle patch containing meloxicam to provide pain management during routine procedures. The goal of this project is to develop and evaluate the use of our engineered microneedle patch in cattle. This goal will be accomplished by evaluating our objectives regarding the efficacy of the engineered patch to manage pain of cattle following a castration procedure, and to assess the long term effect of the patch on cattle growth and behavior. We expect that the microneedle patch will deliver accurate doses of meloxicam for extended pain management. Successful completion of this project will result in novel technology to manage pain in cattle that is easier to implement than current strategies.
Animal Health Component
45%
Research Effort Categories
Basic
10%
Applied
45%
Developmental
45%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3153399102050%
3153399202050%
Goals / Objectives
Delivery of pain medication to animals is most often administered via injections or via oral tablets or suspension. These methods of drug administration often require repeated dosing which is not ideal for livestock owners and veterinarians. This project will study a better approach for long-term drug therapy for cattle. Our goal is to develop a transdermal microneedle patch made from the biodegradable substance chitosan, and evaluate its ability to administer meloxicam, a nonsteriodal anti-inflammatory drug, for pain control in cattle. This technology will deliver a convenient method to minimize or eliminate some of the negative impacts of post-surgical pain caused by procedures such as castration on cattle wellbeing, health and gain performance.Objective 1. To evaluate the efficacy of a novel transdermal microneedle chitosan patch to deliver consistent plasma concentrations of meloxicam to cattle.Objective 2. To evaluate the efficacy of a chitosan microneedle patch containing meloxicam for post-surgical physiologic and inflammatory response in castrated male calves.Objective 3. To evaluate the longterm effects of a meloxicam-chitosan microneedle patch on post-surgical behavior, growth performance and carcass quality when used to control post-surgical pain in male calves castrated at weaning.
Project Methods
This study will evaluate the chitosan needle patch for consistent delivery of meloxicam plasma concentrations as well as evaluate long term pain control and it effects on post-surgical physiologic response, behavioral response, growth performance and carcass quality in castrated male beef calves.Objective 1: In order to evaluate in vivo meloxicam plasma concentrations administered from a chitosan microneedle patch in male calves (n = 6), calves will be allocated randomly to treatments to provide 2 calves per each experimental treatment and 2 calves will serve as a negative control. Experimental treatments will consist of placing a single chitosan microneedle patch containing one of two different dosages of meloxicam on the inside of the calf's left ear. Blood samples will be collected via jugular venipuncture at 0, 20, 40 and 60 minutes; then 2, 4, 7, 10, 24, 30, 48, 72, 96, 120, 144, and 168 hours to evaluate meloxicam levels in blood plasma. The blood plasma samples will be sent to Kansas State University for HPLC and MS analysis to evaluate the two dosages of meloxicam levels in the blood plasma over time.Objective 2: Twenty-four calves will be assigned to 1 of 2 treatments (n = 12 calves/treatment) with the twelve bull calves randomly assigned to receive either a placebo (CON) or meloxicam-chitosan needle patch (MEL) prior to castration. On day one, cattle will be restrained in a squeeze chute for administration of the meloxicam-chitosan needle patches, and at the same time, an indwelling catheter will be placed into the jugular vein of all study animals to facilitate intensive, serial blood collection. Blood samples will be repeatedly collected from all calves at 0, 20, 40 and 60 minutes; then 2, 4, 7, 10, 24, 30, 48, 72, 96, 120, 144, and 168 hours to evaluate substance P, complete blood count, cortisol, prostaglandin E2, haptoglobin, cortisol and meloxicam plasma concentrations.Objective 3: Thirty-six male crossbred Angus calves will be obtained from a local farm and transported to the research unit. Calves will consist of bulls (n = 24) and steers (n = 12). Bull calves being randomly assigned to one of two treatments and the twelve steers serving as a negative control group. Twelve bulls will be randomly assigned to two treatments with twelve receiving a placebo chitosan patch and twelve receiving a meloxicam-chitosan needle patch 1 day prior to surgical castration. The twelve steer calves will serve as negative control (CON). Castration will be performed surgically on the bull calves. Calf behavioral activity will be monitored for the 28-day post-castration period on all calves using a 3-axis accelerometer attached to the right metatarsus using a plastic strap. Accelerometers will be attached to each calf from day 7 through day 28 to observe behavior. The accelerometers will log data every 15 minutes and the variables will include: time spent standing, time spent lying, number of steps taken, number of lying bouts, and a motion index determined from a proprietary algorithm. A baseline value for each behavior variable will be determined for each treatment by determining the behavior variable means recorded from days −7 to −1. During processing and castration on days -7, 0, 7, 14 and 28, animal temperament will be evaluated on each animal while restrained in the hydraulic squeeze chute using a 4-point scale. The evaluator will be masked to drug treatment. Calves will be monitored throughout the entire study for general health and well-being. Individual body weights will be recorded throughout the study on days -7, 0, 7, 14, 28, 56, 84, 112, 140, 168 182 (feedlot entry), interim feedlot weight, and harvest weight. Stalls will be randomly assigned to calves upon arrival, and calves be given a 2-week acclimation period before study commencement. After day 28 of the study, calves will be commingled and moved to a series of twelve 2.4 ha pens and grazed as a single group being rotated through the pastures to supply adequate forage needs. The pens will contain mixed-grass forage and calves will be maintained there for an additional 152-day period to complete the grazing phase of the study. During grazing, the feed management approach will designed to provide cattle with ad libitum hay if forage is limiting and a supplemental commodity ration intended to achieve approximately 0.68 kg of gain/d which will be fed daily. Around day 200, calves will be shipped to a feedlot and fed a total mixed ration until reaching harvest weight. Calves will be assigned to a single feedlot pen and fed a common feedlot ration throughout the finishing period. When steers are determined by feedlot personnel to reach a suitable degree of finish, they will be transported to a commercial processing plant for harvest. Backfat thickness, rib eye area, HCW, marbling score, quality grade, and yield grade data will recorded at harvest of the study calves by a trained technician.

Progress 08/01/23 to 07/31/24

Outputs
Target Audience:We have presented data at the Midwest section meeting of the American Society of Animal Science to target other scientific researchers interested in this topic and area of research. Changes/Problems:Our study objectives were changed and approved last year (2023). We are making progress toward these new objectives. Improvements in the engineering lab to the patch varying the surface area and patch size as well as testing different drug carriers and patch structural polymers are part of the new objectives. We hope these changes will improve the absorption of the meloxicam into the plasma compared to our initial study attempts. What opportunities for training and professional development has the project provided?My PhD student attended the Midwest Section meeting for the American Society of Animal Science in Madison, WI in March 2024 to deliver a presentation regarding our most recent results. She shared an abstract presentation about the most recent work we accomplished sponsored by the NIFA grant. How have the results been disseminated to communities of interest?One abstract will be published in the Journal of Animal Science this year. What do you plan to do during the next reporting period to accomplish the goals?We plan to continue to collect data during a planned study in Fall 2024, and we are preparing a manuscript for publication that we plan to submit later this year (Fall 2024).

Impacts
What was accomplished under these goals? In 2023, we were granted approval to change the original objectives to the following: Revised objectives for USDA-NIFA Grant no. 2021-67015-35675 After facing challenges with maintaining adequate plasma concentration levels of pain medication during preliminary testing, were approved to modify the current objectives to the USDA-NIFA grant 2021-67015-35675. Objective 1: To engineer a microneedle patch with tunable release kinetics by controlling the patch dimensions, drug dosage, and polymer degradation rate. Hypothesis: Controlling the patch dimensions, drug dosage, and degradation rate of the polymer matrix via polymer composition and via chemical crosslinking will yield a microneedle patch with tunable release kinetics allowing accurate dosing of drug over a specified period. Objective 2: To evaluate the in vivo drug release of the two optimized patches in pigs. Hypothesis: Microneedle patches with optimized release kinetics and geometry will result in a patch capable of delivering pain medication to pigs in therapeutic doses. These objectives were evaluated using plasma and ear tissue meloxicam concentrations when the drug was delivered using two different formulations of a microneedle patch placed on the back of the ear. Nursing pigs (n = 10; body weight = 3.64 ± 0.32 kg) from 1 litter were randomly assigned to 1 of 2 treatment groups. Patches delivered 2.5 mg of meloxicam/kg of body weight to pigs regardless of formulation, with the differences in patches being based on type of polymer located on the microneedles and base of the patch. Treatment groups were: 1) pigs receiving a microneedle patch with collagen, polyvinyl alcohol (PVA), and meloxicam on the microneedles, with PVA and chitosan on the base (n = 6 pigs; Patch A); and 2) pigs receiving a microneedle patch with chitosan, PVA, and meloxicam on the microneedles, with collagen and PVA on the base (n = 4 pigs; Patch B). During the study, pigs remained in the crate with the sow and littermates. An Estrotect® Breeding Indicator strip was used to adhere the microneedle patch on the ear of the pigs. Blood was collected at study initiation (0 hours) prior to patch administration and 24 and 48 hours for plasma meloxicam analysis. To evaluate tissue meloxicam concentrations, 3 pigs that received Patch A and 2 pigs that received Patch B were humanely euthanized after blood collection and patch removal at 24 and 48 hours, with the entirety of the ear removed for analysis. Statistical analyses were performed using the MIXED procedure of SAS 9.4 (Cary, NC) to assess the effects of treatment, hour, and treatment × hour interaction. Statistical significance was determined at P ≤ 0.05, with tendencies at 0.05 < P ≤ 0.1. There were no differences (P = 0.83) between patch formulations for tissue meloxicam concentrations (Patch A = 6.55 ± 2.12 ng/mg; Patch B = 7.32 ± 2.60 ng/mg). Plasma concentrations were not detectable at any time point, but tissue concentrations of meloxicam were detectable for pigs that received either patch formulation at 24 (5.44 ± 2.37 ng/mg) and 48 hours (8.42 ± 2.37 ng/mg). There were no differences (P = 0.41) between patch formulations for tissue meloxicam concentrations at either timepoint, nor was there an interaction of patch formulation and hour (P = 0.73). It is unknown why the meloxicam did not diffuse from the ear tissue into the plasma; however, further studies will focus on applying the microneedle patch at different areas on the animal to allow for optimal diffusion through the skin and into blood circulation for pain mitigation in livestock animals.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2024 Citation: Midkiff, K. A., K. A. Miranda-Mu�oz, R. A. Cheek, J. L. Reynolds, T. Tsai, M. Bear, K. P. K. Kompalage, R. Tucker, H. Coetzee, E. B. Kegley, J. Almod�var, J. G. Powell. 2024. Evaluation of a microneedle patch for the delivery of meloxicam in a veterinary application. 2024 ASAS Midwest Section. Madison, WI


Progress 08/01/22 to 07/31/23

Outputs
Target Audience:The target audience represents several stakeholders in the community. Peer reviewed scientific abstracts and manuscripts will be submitted for dissemination of these study findings. Once we have meaningful data, weplan to speak at field days and Extension meetings to relay results of this research project to livestockproducers, county agents and veterinarians. Changes/Problems:In our initial study last year, we have noted that the meloxicam plasma levels were very low, lower than we desired in the animals treated with the microneedle patch. Therefore, we have changed our study objectives this year to make improvements in the engineering lab to the patch and varythe surface area or alter thepolymer and/or drug carrier when the patch is applied to the animal. We hope this will improve absorption of the meloxicam into the plasma compared to what we found on ourinitial study. What opportunities for training and professional development has the project provided?My PhD student attended the Southern Section of the American Society of Animal Science in Raleigh, NC in January 2023 to participate in the conferenceby sharing an abstract presentation about our study sponsored by the NIFA grant,and she enjoyed hearing from other scientists working in the same research area. How have the results been disseminated to communities of interest?One abstract was published in the Journal of Animal Science this past year. What do you plan to do during the next reporting period to accomplish the goals?We plan to continue to collect data, and my student will plan to speak at the Midwest Section meeting of the American Society of Animal Scientists next year (2024) to deliver an abstract presentation to attendees regarding our project summary. We will also plan topublisha journal articlerelated to these research developments in 2024.

Impacts
What was accomplished under these goals? This past year, some challenges arose in our current study. In January, we met virtually with the program leader fromUSDA/NIFA. At this meeting, we were asked to "revise our objectives" for the situation we were facing. The newly revised objectives were reviewed, abd approved by NIFA. Our newly approved objectives are the following: Objective 1: To engineer a microneedle patch with tunable release kinetics by controlling the patch dimensions, drug dosage, and polymer degradation rate. Hypothesis: Controlling the patch dimensions, drug dosage, and degradation rate of the polymer matrix via polymer composition and via chemical crosslinking will yield a microneedle patch with tunable release kinetics allowing accurate dosing of drug over a specified period. Objective 2: To evaluate the in vivo drug release of the two optimized patches in pigs. Hypothesis: Microneedle patches with optimized release kinetics and geometry will result in a patch capable of delivering pain medication to pigs in therapeutic doses.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2023 Citation: Midkiff, K. A., K. M. Munoz, R. Cheek, J. Reynolds, T. C. Tsai, M. Bear. H. Coetzee, E. B. Kegley, J. Almodovar, J. G. Powell. 2023. Evaluation of meloxicam plasma concentrations using a microneedle patch for drug delivery. Southern Section of American Society of Animal Science. Raleigh, NC


Progress 08/01/21 to 07/31/22

Outputs
Target Audience:The target audience represents several stakeholders in the community. Peer reviewed scientific abstracts and manuscripts will be submitted for dissemination of these study findings. Once we have meaningful data, we also plan to speak at field days and Extension meetings to relay results of this research project to beef cattle producers, county agents and veterinarians. Changes/Problems:In our initial study a few months ago, we have noted that the meloxicam plasma levels were lower than we desired in the animals treated with the microneedle patch. Therefore,we have tried to make some improvements in the engineering lab to enlarge the patch and to increase the surface area with the pinna of the ear when applied to the animal. We hope this will improve absorption of the meloxicam into the plasma compared to what we found with this initial study. We plan to restest the improved patch in the Fall 2022. What opportunities for training and professional development has the project provided?My PhDstudentand I plan to attended the Southern Section of the American Society of Animal Science in Raleigh, NC in January 2022 to participate in the conference both by sharing an abstract presentationand hearing from other scientists working in the same research area. How have the results been disseminated to communities of interest?My student willspeak at the Southern Section meeting of the American Society of Animal Scientists to deliver an abstract presentation to attendees regarding our project summaries. We have published a journal articles related to these research developments. What do you plan to do during the next reporting period to accomplish the goals?We will continue to collect data and plan to report results via conference presentations and scientific journal articles.

Impacts
What was accomplished under these goals? We have hired two graduate students in the Fall semester of 2021 to help accomplish the studies that we have planned within this proposal. A PhD student was hired in the ANSC department, and a MS student was hired in CHEG department. They have been instrumental in developing and testing a patch under Objective 1. We conducted a study in the Spring of 2022, and evaluated the plasma concentration of meloxicam using the transdermal microneedle patch in the pinna of the ear.Data were collected and analyzed, and we have planned an abstract presentation reporting the results regarding this objectives.

Publications

  • Type: Journal Articles Status: Published Year Published: 2022 Citation: Castilla-Casadiego, D. A., K. A. Miranda-Munoz, J.L. Roberts, A. D. Crowell, D. Gonzalez-Nino, D. Choudhury, F.O. Aparicio-Solis, S. Servoss, A. M. Rosales, G. Prinz, M.Zou, Y. Zhang, J. F. Coetzee, L. Greenlee, J. G. Powell, J. L. Almodovar. 2022. Biodegradable microneedle patch for delivery of meloxicam for managing pain in cattle. Plos One. 17(8): doi.org/10.1371/journal.pone.0272169
  • Type: Conference Papers and Presentations Status: Submitted Year Published: 2023 Citation: Midkiff, K. A., K. M. Munoz, R. Cheek, J. Reynolds, T. C. Tsai, M. Bear. H. Coetzee, E. B. Kegley, J. Almodovar, J. G. Powell. 2023. Evaluation of meloxicam plasma concentrations using a microneedle patch for drug delivery. Southern Section of American Society of Animal Science. Raleigh, NC