Development of a subcutaneous ear implant to deliver an anaplasmosis vaccine to dairy steers

Bovine anaplasmosis is the most prevalent tick-transmitted disease of cattle worldwide and a major obstacle to profitable beef production. Use of chlortetracycline-medicated feed to control active anaplasmosis infections during the vector season has raised concerns about the potential emergence of antimicrobial resistance in bacteria that may pose a risk to human health. Furthermore, the absence of effectiveness data for a commercially available, conditionally licensed anaplasmosis vaccine is a major impediment to implementing anaplasmosis control programs. The primary objective of this study was to develop a single-dose vaccine delivery platform to produce long-lasting protective immunity against anaplasmosis infections. Twelve Holstein steers, aged 11-12 weeks, were administered a novel 3-stage, single-dose vaccine against Anaplasma marginale (Am) major surface protein 1a. The vaccine consisted of a soluble vaccine administered subcutaneously (s.c.) for immune priming, a vaccine depot of a biodegradable polyanhydride rod with intermediate slow release of the vaccine for boosting immune response, and an immune-isolated vaccine platform for extended antigen release (VPEAR implant) deposited s.c. in the ear. Six calves were randomly assigned to two vaccine constructs (n=3) that featured rods and implants containing a combination of two different adjuvants, diethylaminoethyl (DEAE)-Dextran and Quil-A (Group A). The remaining 6 calves were randomly assigned to two vaccine constructs (n=3) that featured rods and implants containing the same adjuvant (either DEAE-Dextran or Quil A) (Group B). Twenty one months post-implantation, calves were challenged intravenously with Am stabilate and were monitored weekly for signs of fever, decreased packed cell volume (PCV) and bacteremia. Data were analyzed using a mixed effects model and chi-squared tests (SAS v9.04.01, SAS Institute, Cary, NC). Calves in Group A had higher PCV than calves in Group B (P = 0.006) at day 35 post-infection. Calves in Group A were less likely to require antibiotic intervention compared with calves in Group B (P = 0.014). Results indicate that calves exhibited diminished clinical signs of anaplasmosis when antigen was delivered with a combination of adjuvants as opposed to a single adjuvant. This demonstrates the feasibility of providing long lasting protection against clinical bovine anaplasmosis infections using a subcutaneous ear implant vaccine construct

The effect of the anti-inflammatory drug sodium salicylate in mature periparturient dairy cattle and immortalized bovine mammary epitheilal (MAC-T) cells

Master of ScienceDepartment of Animal Sciences and IndustryBarry BradfordDuring the transition period, 3 wk before to 3 wk after calving, dairy cows experience a variety of sudden hormonal and metabolic shifts that could result in metabolic disorders or diseases, which can be detrimental to the productive life and longevity of the cow. Cows undergo a negative energy balance, where they cannot consume enough feed to meet their energy requirements. To make up this deficit, cows mobilize adipose tissue in the form of non-esterified fatty acids (NEFA) which are transported to the liver and are either used for fuel or stored as triglycerides. High levels of circulating NEFA can lead to endoplasmic reticulum (ER) stress, which is linked to inflammation. This low-grade inflammation can compromise cell function. To mitigate this inflammation, sodium salicylate, a non-steroidal anti-inflammatory drug (NSAID), was given to mature (3+ parity) cows for 7 d after parturition via their drinking water. Blood was collected daily and a glucose turnover assay was performed. Liver, muscle, and adipose tissue was collected on d 7. Overall, it appeared that SS increased insulin sensitivity and depressed gluconeogenesis post-transcriptionally. Multiple in vitro studies were performed on immortalized bovine mammary epithelium (MAC-T) cells to determine the action of SS when ER stress was induced with palmitate (PALM). Treatment with SS did not mitigate, and in some cases exacerbated, the ER stress response. The addition of bovine serum albumin (BSA), a common component of cell culture media, may alter reactive oxygen species (ROS) measurements due to its antioxidant property. Overall, SS seems to alter metabolic processes and the cellular response to stress

Comparative Pharmacokinetics and Tissue Concentrations of Flunixin Meglumine and Meloxicam in Tilapia (Oreochromis spp.)

Evidence of pain perception in fish is well established, but analgesic use in aquaculture is limited. The objective was to investigate the comparative pharmacokinetics of flunixin administered intramuscularly (IM) and meloxicam administered IM or orally (PO) in tilapia. Two hundred and seventy fish were assigned to 1 of 3 treatment groups: flunixin meglumine IM (2.2 mg/kg); meloxicam IM (1 mg/kg); or meloxicam PO (1 mg/kg). Blood and tissue samples were collected from 6 fish per treatment at 14 time points for 10 days. Drug concentrations were determined using ultra-high-pressure liquid chromatography coupled with mass spectroscopy. Plasma concentration versus time data were analyzed with a non-compartmental approach using a commercially available software. Flunixin reached a mean maximum concentration (Cmax) of 4826.7 ng/mL at 0.5 h, had a terminal half-life (T1/2) of 7.34 h, and an area under the concentration–time curve extrapolated to infinity (AUCINF_obs) of 25,261.62 h·ng/mL. Meloxicam IM had a T1/2 of 9.4 h after reaching a Cmax of 11.3 ng/mL at 2 h, with an AUCINF_obs of 150.31 h·ng/mL. Meloxicam PO had a T1/2 of 1.9 h after reaching a Cmax of 72.2 ng/mL at 2 h, with an AUCINF_obs of 400.83 h·ng/mL. Tissue concentrations of both drugs were undetectable by 9 h. Flunixin reached a sufficient plasma concentration to potentially have an analgesic effect, while meloxicam, when administered at the given dosage, likely would not

Analgesic Comparison of Flunixin Meglumine or Meloxicam for Soft-Tissue Surgery in Sheep: A Pilot Study

The amount of scientific data evaluating sheep pain responses after analgesia treatment is limited. The aims of this study were to compare the efficacy of flunixin meglumine (FLU) and meloxicam (MEL) at relieving post-surgical pain in sheep and to evaluate the utility of the Sheep Grimace Scale (SGS). Thirty ewes were assigned to one of three treatment groups: oral MEL or intravenous FLU to manage pain associated with a laparotomy procedure, or a non-surgical control (CON) group. Behavior and physiologic outcome measures were collected pre-procedure and up to 48 h post-procedure. There were no significant differences in behavior, gait, degree of inflammation or pain around the surgical site when MEL and FLU sheep were compared, suggesting that both drugs provided similar levels of analgesia. Significant differences in behavior, gait, abdominal inflammation and pain were found when surgical sheep were compared to non-surgical controls. More work is needed to characterize the amount of pain relief provided by MEL and FLU. The SGS had moderate reliability between scorers; however, the results were inconsistent with the other study outcome measures. The SGS may have some utility as a pain assessment tool but should be used in conjunction with other pain measures

Analgesic Comparison of Flunixin Meglumine or Meloxicam for Soft-Tissue Surgery in Sheep: A Pilot Study

The amount of scientific data evaluating sheep pain responses after analgesia treatment is limited. The aims of this study were to compare the efficacy of flunixin meglumine (FLU) and meloxicam (MEL) at relieving post-surgical pain in sheep and to evaluate the utility of the Sheep Grimace Scale (SGS). Thirty ewes were assigned to one of three treatment groups: oral MEL or intravenous FLU to manage pain associated with a laparotomy procedure, or a non-surgical control (CON) group. Behavior and physiologic outcome measures were collected pre-procedure and up to 48 h post-procedure. There were no significant differences in behavior, gait, degree of inflammation or pain around the surgical site when MEL and FLU sheep were compared, suggesting that both drugs provided similar levels of analgesia. Significant differences in behavior, gait, abdominal inflammation and pain were found when surgical sheep were compared to non-surgical controls. More work is needed to characterize the amount of pain relief provided by MEL and FLU. The SGS had moderate reliability between scorers; however, the results were inconsistent with the other study outcome measures. The SGS may have some utility as a pain assessment tool but should be used in conjunction with other pain measures

Delivering an Immunocastration Vaccine via a Novel Subcutaneous Implant

Immunocastration relies on the vaccine-mediated stimulation of an immune response to gonadotropin-releasing hormone (GnRH) in order to interrupt spermatogenesis. This approach offers a less painful alternative to traditional castration approaches but the current, commercially available options require multiple doses of vaccine to maintain sterility. Thus, a series of pilot studies were conducted to determine the feasibility of a single-dose immunocastration vaccine implant. These five studies utilized a total of 44 Holstein bulls to determine the optimal vaccine composition and validate the ability of a stainless-steel subcutaneous implant to deliver a vaccine. Outcome measures included the duration of implant retention, scrotal dimensions and temperature, implant site temperature, anti-GnRH antibodies, and serum testosterone concentration. Over the course of several studies, anti-GnRH antibodies were successfully stimulated by vaccine implants. No significant treatment effects on scrotal dimensions or testosterone were detected over time, but changes in spermatogenesis were detected across treatment groups. Results indicate that a single-dose implantable immunocastration vaccine elicits a humoral immune response and could impact spermatogenesis in bulls. These findings provide opportunities for the refinement of this technology to improve implant retention over longer periods of time. Taken together, this approach will offer producers and veterinarians an alternative to physical castration methods, to improve animal welfare during routine livestock management procedures.This article is published as Curtis, Andrew K., Douglas E. Jones, Michael Kleinhenz, Shawnee Montgomery, Miriam Martin, Mikaela Weeder, Alyssa Leslie et al. "Delivering an Immunocastration Vaccine via a Novel Subcutaneous Implant." Animals 12, no. 19 (2022): 2698. DOI: 10.3390/ani12192698. Copyright 2022 by the authors. Attribution 4.0 International (CC BY 4.0). Posted with permission

Delivering an Immunocastration Vaccine via a Novel Subcutaneous Implant

Immunocastration relies on the vaccine-mediated stimulation of an immune response to gonadotropin-releasing hormone (GnRH) in order to interrupt spermatogenesis. This approach offers a less painful alternative to traditional castration approaches but the current, commercially available options require multiple doses of vaccine to maintain sterility. Thus, a series of pilot studies were conducted to determine the feasibility of a single-dose immunocastration vaccine implant. These five studies utilized a total of 44 Holstein bulls to determine the optimal vaccine composition and validate the ability of a stainless-steel subcutaneous implant to deliver a vaccine. Outcome measures included the duration of implant retention, scrotal dimensions and temperature, implant site temperature, anti-GnRH antibodies, and serum testosterone concentration. Over the course of several studies, anti-GnRH antibodies were successfully stimulated by vaccine implants. No significant treatment effects on scrotal dimensions or testosterone were detected over time, but changes in spermatogenesis were detected across treatment groups. Results indicate that a single-dose implantable immunocastration vaccine elicits a humoral immune response and could impact spermatogenesis in bulls. These findings provide opportunities for the refinement of this technology to improve implant retention over longer periods of time. Taken together, this approach will offer producers and veterinarians an alternative to physical castration methods, to improve animal welfare during routine livestock management procedures

Development of a subcutaneous ear implant to deliver an anaplasmosis vaccine to dairy steers

Bovine anaplasmosis is the most prevalent tick-transmitted disease of cattle worldwide and a major obstacle to profitable beef production. Use of chlortetracycline-medicated feed to control active anaplasmosis infections during the vector season has raised concerns about the potential emergence of antimicrobial resistance in bacteria that may pose a risk to human health. Furthermore, the absence of effectiveness data for a commercially available, conditionally licensed anaplasmosis vaccine is a major impediment to implementing anaplasmosis control programs. The primary objective of this study was to develop a single-dose vaccine delivery platform to produce long-lasting protective immunity against anaplasmosis infections. Twelve Holstein steers, aged 11-12 weeks, were administered a novel 3-stage, single-dose vaccine against Anaplasma marginale (Am) major surface protein 1a. The vaccine consisted of a soluble vaccine administered subcutaneously (s.c.) for immune priming, a vaccine depot of a biodegradable polyanhydride rod with intermediate slow release of the vaccine for boosting immune response, and an immune-isolated vaccine platform for extended antigen release (VPEAR implant) deposited s.c. in the ear. Six calves were randomly assigned to two vaccine constructs (n=3) that featured rods and implants containing a combination of two different adjuvants, diethylaminoethyl (DEAE)-Dextran and Quil-A (Group A). The remaining 6 calves were randomly assigned to two vaccine constructs (n=3) that featured rods and implants containing the same adjuvant (either DEAE-Dextran or Quil A) (Group B). Twenty one months post-implantation, calves were challenged intravenously with Am stabilate and were monitored weekly for signs of fever, decreased packed cell volume (PCV) and bacteremia. Data were analyzed using a mixed effects model and chi-squared tests (SAS v9.04.01, SAS Institute, Cary, NC). Calves in Group A had higher PCV than calves in Group B (P = 0.006) at day 35 post-infection. Calves in Group A were less likely to require antibiotic intervention compared with calves in Group B (P = 0.014). Results indicate that calves exhibited diminished clinical signs of anaplasmosis when antigen was delivered with a combination of adjuvants as opposed to a single adjuvant. This demonstrates the feasibility of providing long lasting protection against clinical bovine anaplasmosis infections using a subcutaneous ear implant vaccine construct.This is a pre-copyedited, author-produced version of an article accepted for publication in Journal of Animal Science. The version of record: Curtis, Andrew K., Kathryn E. Reif, Michael D. Kleinhenz, Miriam S. Martin, Brandt Skinner, Sean M. Kelly, Douglas E. Jones, Emily J. Reppert, Shawnee R. Montgomery, Balaji Narasimhan, Tippawan Anantatat, Majid Jaberi-Douraki, and Johann F. Coetzee. "Development of a subcutaneous ear implant to deliver an anaplasmosis vaccine to dairy steers." Journal of Animal Science (2019) is available online at DOI: 10.1093/jas/skz392. Posted with permission.</p