Refinement of water-based foam depopulation procedures for finisher pigs during field conditions: Welfare implications and logistical aspects.

Water-based foam (WBF) depopulation is currently being researched as an alternative for rapid destruction of swine populations under emergency circumstances. Appropriate guidelines are needed to maintain method reliability and depopulation efficacy while minimizing animal distress under field conditions. Finisher pigs were depopulated using WBF with a 7.5-minute dwell time in two trials to evaluate the effect of; trial 1) foam fill level (1.5, 1.75, or 2.0 times the pig's head height) and trial 2) foam fill rate (slow, medium, or fast) on aversive pig responses (surface breaks, vocalization, and escape attempts) and time to cessation of cardiac activity. Activity and cardiac activity were recorded using subcutaneous bio-loggers for swine in trial 2. The average time to cessation of movement (COM) from the start of foam filling was compared for the foam fill rate groups using a generalized linear mixed effect model under Poisson distribution. Foam rate group was used as an independent variable, and replicates as a random effect. For trial 1, the average (mm:ss ± SD) time to fill completion was 01:18 ± 00:00, 00:47 ± 00:05, and 00:54 ± 00:05, for 1.5, 1.75, and 2.0 times the pig's head height, respectively. For trial 2, the average time to fill completion was 03:57 ± 00:32, 01:14 ± 00:23 and 00:44 ± 00:03, and the average time (mm:ss ± SE) to COM was 05:22 ± 00:21, 03:32 ± 00:14, and 03:11 ± 00:13 for slow, medium, and fast fill rate groups, respectively. A higher number of aversive pig responses were observed for the lowest foam fill level and slowest foam fill rate compared to increased fill levels and faster fill rates. For trial 2 the median (mm:ss ± IQR) time to fatal arrhythmia was 09:53 ± 02:48, 11:19 ± 04:04, and 10:57 ± 00:47 post-foam initiation for fast, medium, and slow foam rate groups, respectively. Time to cessation of cardiac activity was significantly shorter for the fast foam rate group compared to medium and slow foam rates groups (P = 0.04). For both trials, vocalizations were absent, and all pigs were unconscious following the 7.5-minute dwell time and no pigs needed a secondary euthanasia method. This WBF study showed that slower fill rates and low foam fill levels may extend the time until cessation of cardiac activity in swine during depopulation. A conservative recommendation with consideration of swine welfare during an emergency scenario would be a minimum foam fill level twice the pig's head height and a foam fill rate capable of covering pigs in foam within 60 s to minimize aversive responses and expedite cessation of cardiac activity.

Water-based medium-expansion foam depopulation of adult cattle.

Current options for depopulation of adult cattle are limited, have logistic constraints, and may not be practical on a large scale. Aspirated water-based foam (WBF) has been shown to be successful in depopulating poultry and swine but has yet to be tested in cattle. WBF is advantageous because necessary equipment can be readily available, easy to use, and presents minimal personnel risk. With the use of a modified rendering trailer in a field setting, we evaluated the efficacy of aspirated WBF for depopulation of adult cattle. Water-based medium-expansion foam was added to the trailer holding cattle to a depth of approximately 50 cm greater than head height. The study was conducted as a gated design and the initial trial was conducted using six anesthetized and six conscious animals for verification of the process and followed by four replicates each containing 18 conscious cattle. A total of 84 cattle were used, with a subset (n = 52) implanted with subcutaneous bio-loggers that recorded activity and electrocardiograms. Cattle were loaded onto the trailer and three gasoline-powered water pumps delivered foam into the trailer followed by a 15-min foam dwell period. Average (± SD) time to completely fill the trailer with foam was 84.8 ±â€…11.0 s. No animal vocalizations were heard during foam application or the dwell period, and all cattle were confirmed dead upon removal from the trailer after 15 min of immersion. Necropsies of a subset of cattle revealed foam extending to at least the tracheal bifurcation in all cattle and distal to this level in 67% (8/12) animals. Time to cessation of movement, which served as a proxy for loss of consciousness, was 2.5 ±â€…1.3 min and time to cardiac death was 8.5 ±â€…2.5 min as determined by data from animals carrying subcutaneous bio-loggers. The results of this study indicate that WBF is a rapid and effective method for depopulation of adult cattle with potential advantages in speed and carcass handling and disposal over current methods.

Mechanical Cardiac Support With an Implantable Direct Cardiac Compression Device: Proof of Concept.

PURPOSE: We examined the hemodynamic effects of a new, implantable, direct cardiac assist device in an ovine heart failure model. DESCRIPTION: The device, which encompasses both left and right ventricles, is inserted through the pericardial apex and self-expands to encompass the heart without suturing. The intact pericardium anchors the device in place. The device has 2 concentric chamber layers: an internal chamber layer filled with fluid to conform to the heart and an external chamber layer filled with air that provides external compression and negative pressure to aid relaxation. EVALUATION: The device was implanted in 7 sheep with heart failure induced by microsphere embolization. Cardiac performance was assessed for 6 to 8 hours. The cardiac assist device provided cardiac systolic and diastolic assistance, as shown by pressure tracings of the left ventricle and aorta, pulmonary artery flow, and +dP/dt. Central venous pressure decreased during cardiac assistance. No anatomic damage was noted postmortem. CONCLUSIONS: Systolic and diastolic cardiac assistance can be achieved with this device that compresses and relaxes in synchrony with the native cardiac cycle.

Reliability of water-based medium-expansion foam as a depopulation method for nursery pigs and cull sows.

Animal disease preparedness plans including depopulation guidelines are paramount to prevent the spread of emerging infectious diseases but difficult to implement for swine under field conditions. However, water-based foam (WBF) is currently an approved and successfully deployed depopulation methodology in poultry. Therefore, the reliability of WBF as a depopulation method and the effectiveness and irreversibility of consciousness and consequential mortality in pigs of different ages was assessed across two trials. Trial 1 investigated the time to loss of consciousness and cessation of cardiac activity in nursery pigs (n = 72) at six different foam immersion time points (2.5, 5, 7.5, 10, 12.5 and 15 min) when placed in a 1.47 m3 (1.2 × 1.2 × 1.02 m, length × width × height) plastic bulk container. One pig per replicate was implanted with an ECG bio-logger. Irreversible loss of consciousness was observed after a 5-min immersion. The average (SD) time to development of a fatal arrhythmia from the initiation of the foam application was 7.3 min (1.82 s). Trial 2 aimed to validate the findings from Trial 1 in 75 larger cull sows across three replicates (n = 25). Sows were loaded into a 41-m3 sealed trailer (12.2 × 1.5 × 2.24 m), immersed in WBF and left undisturbed for 5 min post foam-filling completion. Six pigs in each replicate were implanted with an ECG bio-logger. A 5-min dwell time resulted in irreversible loss of consciousness and subsequent mortality in all cull sows. The average time (SD) to cessation of movement and fatal arrhythmia post foam-filling completion was 2.2 min (34.8 s) and 8.7 min (138.0 s), respectively. While a 5-min immersion in WBF induced irreversible loss of consciousness and death in both trials, a 7.5-min dwell time followed by observation for confirmation of death post WBF removal would be advisable for pigs of all sizes.

Evaluation of a Water-Based Medium-Expansion Foam Depopulation Method in Suckling and Finisher Pigs.

The threat of foreign animal disease introduction through contaminated animal products, feed ingredients, and wildlife vectors have highlighted the need for additional approved methods for mass depopulation of swine under emergency scenarios, especially methods that can be applied to pigs across all production phases. The market disruption within the swine industry due to the SARS-CoV-2 pandemic has demonstrated this lack of preparation. The objective of this study was to validate water-based foam as a mass depopulation method for suckling (18 to 24 days of age) and finisher stage (63 to 100 days of age) pigs. Finisher pigs (n = 31, originally 32 but one finisher pig died prior to foaming), allocated as 9 triads and 1 set of 4 pigs, in 10 total replicates, and suckling pigs (n = 32), randomly allocated to two replicates, were completely covered in water-based medium-expansion foam for a 15-min dwell time in a bulk container. Container fill time for the trials were 6.5 ± 0.68 s and 5.3 ± 0.03 s for finisher and suckling pig replicates, respectively. Average (± SD) time for cessation of movement was 105 ± 39.1 s (s) for finisher pigs and 79.5 ± 10.5 s for suckling pigs. After completion of the 15-min dwell time in the foam, all pigs were confirmed dead upon removal from the container. The results from the present study suggest that the use of water-based foam can be an effective means of mass depopulation for suckling and finisher stage pigs, supporting previous research on the application to adult swine.

Continual cerebrospinal fluid sampling in the neonatal domestic piglet for biomarker and discovery studies.

BACKGROUND: Longitudinal access to cerebrospinal fluid (CSF) is useful for biomarker discovery in neurological disorders or diseases affecting CSF composition. Here, we aim to test a new method for insertion of a permanent intrathecal catheter, facilitating longitudinal collection of CSF. NEW METHOD: We surgically placed a permanent intrathecal catheter into the cisterna magna of anesthetized neonatal piglets. The thecal sac was accessed at the L5-S1 spinal level and a radiopaque catheter was inserted under fluoroscopic x-ray guidance to position the tip at the cisterna magna. A titanium access port was connected to the catheter and anchored subcutaneously. Immediately after surgery, we confirmed CSF flow through the catheter and port via needle aspiration. Catheter patency over a two-month study period was determined through periodic CSF collection from the port. RESULTS: Frequent (up to 3 times weekly), longitudinal sampling of CSF was achievable in neonatal piglets up to 60 days after implantation. CSF was readily accessible through the port without major adverse events. Catheterized piglets demonstrated slower, but normal, weight gain compared to control piglets. Post-operative complications were managed with standard access precautions and medications. There were no complications involving the implanted hardware. COMPARISON WITH EXISTING METHOD(S): This method fills a critical gap in the existing methods for longitudinal CSF sampling through an implanted intrathecal catheter system in neonatal piglets. CONCLUSIONS: This novel method is both safe and effective for longitudinal CSF access in the domestic piglet. Catheter patency and access to CSF is maintained over multiple months without major adverse events.

Direct Interstitial Decongestion in an Animal Model of Acute-on-Chronic Ischemic Heart Failure.

Removal of excess fluid in acute decompensated heart failure (ADHF) targets the intravascular space, whereas most fluid resides in the interstitial space. The authors evaluated an approach to interstitial decongestion using a device to enhance lymph flow. The device was deployed in sheep with induced heart failure (HF) and acute volume overload to create a low-pressure zone at the thoracic duct outlet. Treatment decreased extravascular lung water (EVLW) volume (mL/kg) (-32% ± 9%, P = 0.029) compared to controls (+46% ± 9%, P = 0.003). Device-mediated thoracic duct decompression effectively reduced EVLW. Human studies may establish device-based interstitial decongestion as a new ADHF treatment.

Comparison of Gaseous and Water-Based Medium-Expansion Foam Depopulation Methods in Cull Sows.

The U.S. swine industry is currently inadequately prepared to counteract the increasing threat of high-consequence diseases. Although approved and preferred depopulation guidelines exist, ventilation shutdown (VSD+) is currently the only method being deployed during a state of emergency to depopulate large swine populations. However, the permitted use of VSD+ during constrained circumstances has been criticized due to raised swine welfare concerns. The objective of this study was to investigate the effectiveness of carbon dioxide gas (CO2), nitrogen gas (N2), compressed air foam (CAF), compressed nitrogen foam (CAF-N2) and aspirated foam (AF) during a 15-min dwell time on adult swine in an emergency depopulation situation. A small-scale trial using 12 sows per depopulation method showed the highest efficiency to induce cessation of movement for AF and CO2 (186.0 ± 48 vs. 202.0 ± 41, s ± SD). The ease of implementation and safety favored AF for further investigation. A large-scale field study using AF to depopulate 134 sows in modified rendering trailers showed a mean fill time of 103.8 s (SD: 5.0 s) and cessation of movement of 128.0 s (SD: 18.6 s) post filling. All sows were confirmed dead post-treatment for both trials. The implementation of AF in modified rendering trailers may allow for a safe and reliable method that allows for the expedient and mobile depopulation of both small and large numbers of sows during an emergency.

Bilateral Arteriovenous Shunts as a Method for Evaluating Tissue-Engineered Vascular Grafts in Large Animal Models.

There remains a need for large animal models to evaluate tissue-engineered vascular grafts (TEVGs) under arterial pressure to provide preclinical data for future potential human clinical trials. We present a comprehensive method for the interrogation of TEVGs, using an ovine bilateral arteriovenous (AV) shunt implantation model. Our results demonstrate that this method can be performed safely without complications, specifically acute heart failure, steal syndrome, and hypoxic brain injury, and it is a viable experimental paradigm. Our method allows for a non-invasive evaluation of TEVGs in terms of graft flow, graft diameter, and graft patency, while also allowing for graft needle puncture under ultrasound guidance. In addition, traditional pathological analysis, histology, and immunohistochemistry may be performed with the contralateral side providing paired control data to eliminate inter-subject variability while reducing the total number of animals. Further, we present a review of existing literature of preclinical evaluation of TEVGs in large animal models as AV conduits.

Short-term effects of oral dronedarone administration on cardiac function, blood pressure and electrocardiogram in conscious telemetry dogs.

Dronedarone is a multichannel blocking antiarrhythmic drug that has been used for management of atrial fibrillation in humans, but the data in veterinary medicine are inadequate. The objective of this study was to determine the short-term effects of oral dronedarone on cardiac inotropy and lusitropy, blood pressure and electrocardiogram (ECG) in healthy dogs. A total of 6 beagle dogs were instrumented with telemetry units and sono-micrometry crystals to obtain left ventricular pressure-volume relationship, mean blood pressure (MBP) and ECG. Dogs were given orally dronedarone (20 mg/kg, twice per day) for 7 days. All parameters were obtained hourly at 4-8 hr after the first dose and at 12-, 96- (day 4) and 168-hr (day 7) after dosing. The results showed that dronedarone had no effect on inotropy and lusitropy, while it significantly lengthened PQ interval (P<0.001) and lowered MBP (P<0.05). Dronedarone also tended to reduce cardiac output (P=0.237) and heart rate (P=0.057). These results suggested that short-term effects of oral dronedarone administration at a dose of 20 mg/kg, twice per day, produced negative dromotropy with minimal effect on cardiac function in conscious dogs.

Cardiac function after acute support with direct mechanical ventricular actuation in chronic heart failure.

Direct mechanical ventricular actuation (DMVA) exerts direct cardiac compression/decompression and does not require blood contact. The safety and effects of DMVA support in chronically dysfunctional beating hearts in vivo have not been established. This study evaluated hemodynamics and load-independent systolic/diastolic cardiac function before/after acute support (2 hours) using DMVA in small hearts with induced chronic failure. Chronic heart failure was created in seven small dogs (15 ± 2 kg) via either serial coronary microembolizations or right-ventricular overdrive pacing. Dogs were instrumented to measure cardiac output, hemodynamic pressures, left ventricular volumes for pressure-volume analysis via preload reduction. Temporary cardiac support using a DMVA device was instituted for 2 hours. Hemodynamic and mechanical assessments, including dobutamine dose-responses, were compared both before and after support. Hemodynamic indices were preserved with support. Both left-ventricular systolic and diastolic function were improved postsupport, as the slopes of the preload-recruitable stroke work (+29 ± 7%, p < 0.05) and the end-diastolic pressure-volume relationship (EDPVR: -28 ± 9%, p < 0.05) improved post-DMVA support. Diastolic/systolic myocardial reserve, as assessed by responsiveness to dobutamine challenges, was preserved after DMVA support. Short-term DMVA support can safely and effectively sustain hemodynamics, whereas triggering favorable effects on cardiac function in the setting of chronic heart failure. In particular, DMVA support preserved load-independent diastolic function and reserve.

Fused piperidines as a novel class of potent and orally available transient receptor potential melastatin type 8 (TRPM8) antagonists.

The transient receptor potential melastatin type 8 (TRPM8) is a nonselective cation channel primarily expressed in a subpopulation of sensory neurons that can be activated by a wide range of stimuli, including menthol, icilin, and cold temperatures (<25 °C). Antagonism of TRPM8 is currently under investigation as a new approach for the treatment of pain. As a result of our screening efforts, we identified tetrahydrothienopyridine 4 as an inhibitor of icilin-induced calcium influx in CHO cells expressing recombinant rat TRPM8. Exploration of the structure-activity relationships of 4 led to the identification of a potent and orally bioavailable TRPM8 antagonist, tetrahydroisoquinoline 87. Compound 87 demonstrated target coverage in vivo after oral administration in a rat pharmacodynamic model measuring the prevention of icilin-induced wet-dog shakes (WDS).

Antihyperalgesic effects of (R,E)-N-(2-hydroxy-2,3-dihydro-1H-inden-4-yl)-3-(2-(piperidin-1-yl)-4-(trifluoromethyl)phenyl)-acrylamide (AMG8562), a novel transient receptor potential vanilloid type 1 modulator that does not cause hyperthermia in rats.

Antagonists of the vanilloid receptor TRPV1 (transient receptor potential vanilloid type 1) have been reported to produce antihyperalgesic effects in animal models of pain. These antagonists, however, also caused concomitant hyperthermia in rodents, dogs, monkeys, and humans. Antagonist-induced hyperthermia was not observed in TRPV1 knockout mice, suggesting that the hyperthermic effect is exclusively mediated through TRPV1. Since antagonist-induced hyperthermia is considered a hurdle for developing TRPV1 antagonists as therapeutics, we investigated the possibility of eliminating hyperthermia while maintaining antihyperalgesia. Here, we report four potent and selective TRPV1 modulators with unique in vitro pharmacology profiles (profiles A through D) and their respective effects on body temperature. We found that profile C modulator, (R,E)-N-(2-hydroxy-2,3-dihydro-1H-inden-4-yl)-3-(2-(piperidin-1-yl)-4-(trifluoromethyl)phenyl)acrylamide (AMG8562), blocks capsaicin activation of TRPV1, does not affect heat activation of TRPV1, potentiates pH 5 activation of TRPV1 in vitro, and does not cause hyperthermia in vivo in rats. We further profiled AMG8562 in an on-target (agonist) challenge model, rodent pain models, and tested for its side effects. We show that AMG8562 significantly blocks capsaicin-induced flinching behavior, produces statistically significant efficacy in complete Freund's adjuvant- and skin incision-induced thermal hyperalgesia, and acetic acid-induced writhing models, with no profound effects on locomotor activity. Based on the data shown here, we conclude that it is feasible to modulate TRPV1 in a manner that does not cause hyperthermia while maintaining efficacy in rodent pain models.

The vanilloid receptor TRPV1 is tonically activated in vivo and involved in body temperature regulation.

The vanilloid receptor TRPV1 (transient receptor potential vanilloid 1) is a cation channel that serves as a polymodal detector of pain-producing stimuli such as capsaicin, protons (pH <5.7), and heat. TRPV1 antagonists block pain behaviors in rodent models of inflammatory, neuropathic, and cancer pain, suggesting their utility as analgesics. Here, we report that TRPV1 antagonists representing various chemotypes cause an increase in body temperature (hyperthermia), identifying a potential issue for their clinical development. Peripheral restriction of antagonists did not eliminate hyperthermia, suggesting that the site of action is predominantly outside of the blood-brain barrier. Antagonists that are ineffective against proton activation also caused hyperthermia, indicating that blocking capsaicin and heat activation of TRPV1 is sufficient to produce hyperthermia. All TRPV1 antagonists evaluated here caused hyperthermia, suggesting that TRPV1 is tonically activated in vivo and that TRPV1 antagonism and hyperthermia are not separable. TRPV1 antagonists caused hyperthermia in multiple species (rats, dogs, and monkeys), demonstrating that TRPV1 function in thermoregulation is conserved from rodents to primates. Together, these results indicate that tonic TRPV1 activation regulates body temperature.