Use Of Large Animals Research Articles

A Novel Model of Staphylococcus aureus-Induced Lymphoplasmacytic Rhinosinusitis in Rats.

Chronic rhinosinusitis (CRS) is characterized by sinonasal mucosal inflammation. Staphylococcus aureus (S. aureus) is associated with severe CRS phenotypes. Different animal models have been proposed to study the association of CRS and S. aureus. However, current animal models are expensive due to the use of large animals, have high barriers to ethics approval, or require invasive surgical intervention, necessitating a need for a model that can overcome these limitations. This study aimed at establishing a reliable and efficient rat lymphoplasmacytic inflammatory model for rhinosinusitis. Sprague Dawley rats received a daily intranasal application of 20 μL of saline, S. aureus CI-182 exoprotein (250 μg/mL), or exoprotein CI-182 in combination with S. aureus clinical isolate (CI-908 or CI-913) 108 colony-forming unit (CFU)/mL. The rats' sinuses were harvested at 1 and 2 weeks post-intervention. The CFU and histopathologic examination of inflammation were evaluated. S. aureus clinical isolates CI-908 or CI-913 in combination with the exoprotein (CI-182) had higher CFUs and caused persistently higher inflammation at both the 1 and 2-week post-intervention compared to the exoprotein and saline group. The observed inflammatory cell type was lymphoplasmacytic. This study provided evidence that the combination of a S. aureus exoprotein with S. aureus induces inflammation that persists for a minimum of two weeks post-intervention. This model is the first known animal model to create the lymphoplasmacytic inflammation subtype seen in CRS patients. This offers a cost-effective, accessible, non-invasive, and easy-to-replicate model to study the causes and treatment of such inflammation.

A practical guide to setting up pig models for cardiovascular catheterization, electrophysiological assessment and heart disease research.

Over the past years, the use of large animals has become increasingly interesting in translational research, to bridge the gap between basic research in rodents and targeted therapies in humans. Pigs are highly valued in cardiovascular research because of their anatomical, hemodynamic and electrophysiological features, which closely resemble those of humans. For studying these aspects in swine, cardiac catheterization techniques are essential procedures. Although cardiac catheterization seems to be comparatively easy in pigs as human equipment can be used to perform the procedure, there are some pitfalls. Here we provide a detailed protocol to guide the reader through different aspects of cardiac catheterization in pigs. We suggest an approach for safe intubation and extubation, provide tips for perioperative and postoperative management of the animals and guide the reader through different experimental steps, including sheath insertion. We also describe the procedures for basic electrophysiological assessment of conduction properties and atrial fibrillation induction, hemodynamic assessment via pressure-volume loops, right heart and left heart catheterization and the development of a myocardial infarction model by balloon occlusion. This protocol was developed in Landrace pigs and can be adapted to other pig breeds or other large animal species. This protocol requires approximately six and a half working hours in total and should be performed by researchers with previous experience in large animal experimentation and in the presence of a veterinarian.

Modeling the gene delivery process of the needle array-based tissue nanotransfection.

Hollow needle array-based tissue nanotransfection (TNT) presents an in vivo transfection approach that directly translocate exogeneous genes to target tissues by using electric pulses. In this work, the gene delivery process of TNT was simulated and experimentally validated. We adopted the asymptotic method and cell-array-based model to investigate the electroporation behaviors of cells within the skin structure. The distribution of nonuniform electric field across the skin results in various electroporation behavior for each cell. Cells underneath the hollow microchannels of the needle exhibited the highest total pore numbers compared to others due to the stronger localized electric field. The percentage of electroporated cells within the skin structure, with pore radius over 10 nm, increases from 25% to 82% as the applied voltage increases from 100 to 150 V/mm. Furthermore, the gene delivery behavior across the skin tissue was investigated through the multilayer-stack-based model. The delivery distance increased nonlinearly as the applied voltage and pulse number increased, which mainly depends on the diffusion characteristics and electric conductivity of each layer. It was also found that the skin is required to be exfoliated prior to the TNT procedure to enhance the delivery depth. This work provides the foundation for transition from the study of murine skin to translation use in large animals and human settings.

A Wirelessly Controlled Scalable 3D-Printed Microsystem for Drug Delivery.

Here we present a 3D-printed, wirelessly controlled microsystem for drug delivery, comprising a refillable microreservoir and a phase-change peristaltic micropump. The micropump structure was inkjet-printed on the back of a printed circuit board around a catheter microtubing. The enclosure of the microsystem was fabricated using stereolithography 3D printing, with an embedded microreservoir structure and integrated micropump. In one configuration, the microsystem was optimized for murine inner ear drug delivery with an overall size of 19 × 13 × 3 mm3. Benchtop results confirmed the performance of the device for reliable drug delivery. The suitability of the device for long-term subcutaneous implantation was confirmed with favorable results of implantation of a microsystem in a mouse for six months. The drug delivery was evaluated in vivo by implanting four different microsystems in four mice, while the outlet microtubing was implanted into the round window membrane niche for infusion of a known ototoxic compound (sodium salicylate) at 50 nL/min for 20 min. Real-time shifts in distortion product otoacoustic emission thresholds and amplitudes were measured during the infusion, demonstrating similar results with syringe pump infusion. Although demonstrated for one application, this low-cost design and fabrication methodology is scalable for use in larger animals and humans for different clinical applications/delivery sites.

Mouse model as an efficacy test for foot-and-mouth disease vaccines.

Protection against foot-and-mouth disease virus (FMDV) has been linked to the development of a humoral response. In Argentina, the official control tests for assessing the potency of FMD vaccines are protection against podal generalization (PPG) and expected percentage of protection (EPP) curves built with quantitative data of antibodies determined by liquid-phase blocking ELISA (lpELISA). The results of these tests are used to accept or discard vaccines at the batch level. In this report, a mouse model was assessed as an alternative efficacy control for FMDV vaccines. To this aim, groups of cattle (n=18) and BALB/c mice (n=16) were inoculated with commercial FMDV vaccines and bleedings were performed 60days post vaccination (dpv) in cattle and 21dpv in mice. Specific FMDV antibody titres were measured in both species by a standardized lpELISA. A statistically significant association between antibody levels in cattle and mice has already been demonstrated. However, some vaccines have been misclassified since they were considered protective based on lpELISA results but did not induce good protection in cattle upon challenge. For this reason, other immunological parameters were evaluated to improve the prediction of protection in mice, without the need of using infective virus. In addition, antibody titres by lpELISA, the IgG2b/IgG1 isotype ratio and the Avidity Index were identified as good predictors, resulting in an optimal predictive model of protection. This mouse model could be a simple and economic alternative for testing FMD vaccines since the disadvantages of high costs and facility requirements associated with the use of large animals are overcome.

The use of large animals to facilitate the process of MSC going from laboratory to patient\u2014\u2018bench to bedside\u2019

Large animal models have been widely used to facilitate the translation of mesenchymal stem cells (MSC) from the laboratory to patient. MSC, with their multi-potent capacity, have been proposed to have therapeutic benefits in a number of pathological conditions. Laboratory studies allow the investigation of cellular and molecular interactions, while small animal models allow initial ‘proof of concept’ experiments. Large animals (dogs, pigs, sheep, goats and horses) are more similar physiologically and structurally to man. These models have allowed clinically relevant assessments of safety, efficacy and dosing of different MSC sources prior to clinical trials. In this review, we recapitulate the use of large animal models to facilitate the use of MSC to treat myocardial infarction—an example of one large animal model being considered the ‘gold standard’ for research and osteoarthritis—an example of the complexities of using different large animal models in a multifactorial disease. These examples show how large animals can provide a research platform that can be used to evaluate the value of cell-based therapies and facilitate the process of ‘bench to bedside’.

A nanoliter resolution implantable micropump for murine inner ear drug delivery

Advances in protective and restorative biotherapies have created new opportunities to use site-directed, programmable drug delivery systems to treat auditory and vestibular disorders. Successful therapy development that leverages the transgenic, knock-in, and knock-out variants of mouse models of human disease requires advanced microsystems specifically designed to function with nanoliter precision and with system volumes suitable for implantation. Here we present results for a novel biocompatible, implantable, scalable, and wirelessly controlled peristaltic micropump. The micropump configuration included commercially available catheter microtubing (250 μm OD, 125 μm ID) that provided a biocompatible leak-free flow path while avoiding complicated microfluidic interconnects. Peristaltic pumping was achieved by sequentially compressing the microtubing via expansion and contraction of a thermal phase-change material located in three chambers integrated adjacent to the microtubing. Direct-write micro-scale printing technology was used to build the mechanical components of the micropump around the microtubing directly on the back of a printed circuit board assembly (PCBA). The custom PCBA was fabricated using standard commercial processes providing microprocessor control of actuation and Bluetooth wireless communication through an Android application. The results of in vitro characterization indicated that nanoliter resolution control over the desired flow rates of 10–100 nL/min was obtained by changing the actuation frequency. Applying 10× greater than physiological backpressures and ± 3 °C ambient temperature variation did not significantly affect flow rates. Three different micropumps were tested on six mice for in vivo implantation of the catheter microtubing into the round window membrane niche for infusion of a known ototoxic compound (sodium salicylate) at 50 nL/min for 20 min. Real-time shifts in distortion product otoacoustic emission thresholds and amplitudes were measured during the infusion. There were systematic increases in distortion product threshold shifts during the 20-min perfusions; the mean shift was 15 dB for the most basal region. A biocompatibility study was performed to evaluate material suitability for chronic subcutaneous implantation and clinical translational development. The results indicated that the micropump components successfully passed key biocompatibility tests. A micropump prototype was implanted for one month without development of inflammation or infection. Although tested here on the small murine cochlea, this low-cost design and fabrication methodology is scalable for use in larger animals and for clinical applications in children and adults by appropriate scaling of the microtubing diameter and actuator volume.

ROLE OF ANIMAL MODELS IN PERIODONTAL RESEARCH - A REVIEW

Periodontal diseases require treatment at an early stage to prevent further damage and aggravation of the disease. The most commonly seen periodontal diseases are gingivitis and periodontitis. Animals have contributed a major role in studying the different periodontal diseases and providing a proper treatment. Periodontal diseases are either induced in these experimental animal models or can be seen naturally. Different drugs are tested on the animals induced by the disease to find the most effective treatment for that particular disease. Different animals such as mice, rats, pigs, rabbits, hamsters, and rodents are used for the periodontal research. Different animals show a different reaction while some animals show no reaction. Each animal has its own advantages and disadvantages. The use of large animals brings a limitation in the due to its housing difficulties. Animals for periodontal research are chosen depending on their similarity with that of human anatomy and physiology. The use of these animals will help to replicate the disease seen in humans in a better and more accurate way. This will improve the treatment outcome and the prognosis of the disease. The drugs used can, hence, give a better idea about the effect it would have on the human body depending on the effects it shows on the animal models. Hence, the use of appropriate animals for the periodontal research is important to design a better treatment for these diseases. Hence, animal models play an important role in the periodontal research.

Seizure-like activity after gadolinium administration in an anaesthetised foal undergoing magnetic resonance imaging

The use of MRI has been recently increasing in veterinary patients, and this modality has been adapted for use in large animals with the concurrent use of gadolinium-based contrast media....

Contribution of Large Animals to Translational Research on Prenatal Programming of Obesity and Associated Diseases.

The awareness of factors causing obesity and associated disorders has grown up in the last years from genome to a more complicated concept (developmental programming) in which prenatal and early-postnatal conditions markedly modify the phenotype and homeostasis of the individuals and determine juvenile growth, life-time fitness/obesity and disease risks. Experimentation in human beings is impeded by ethical issues plus inherent high variability and confounding factors (genetics, lifestyle and socioeconomic heterogeneity) and preclinical studies in adequate translational animal models are therefore decisive. Most of the studies have been performed in rodents, whilst the use of large animals is scarce. Having in mind body-size, handlingeasiness and cost-efficiency, the main large animal species for use in biomedical research are rabbits, sheep and swine. The choice of the model depends on the research objectives. To outline the main features of the use of rabbits, sheep and swine and their contributions as translational models in prenatal programming of obesity and associated disorders.

Empowering Translational Research in Fetal Growth Restriction: Sheep and Swine Animal Models

Fetal or intrauterine growth restriction (FGR or IUGR) is a concerning health issue not only due to its implications in mortality and morbidity of neonates but also because of its long-term consequences on health and disease risk of the individuals. Its main cause is an insufficient supply of nutrients and oxygen by maternal (malnutrition or hypobaric hypoxia) or placental factors (placental insufficiency) during late gestation, when the requirements of fetus are higher. The availability of reliable animal models would be highly useful for the future development of diagnostic, preventive and therapeutic strategies. Most of the studies using animal models have been performed in rodents, while the use of large animals (sheep and swine) has been scarce. The objective of the current review is to offer an overview on the possibilities of using large animals for conducting translational research on IUGR related to inadequate maternal conditions and/or placental dysfunction.

Chemotherapy-Induced Peripheral Neuropathic Pain and Rodent Models.

Painful peripheral neuropathies resulting from cancer chemotherapy treatment is frequently dose-dependent and may diminish following dose reduction or termination of chemotherapy. However, dose reduction or treatment termination could lead to reemergence of the cancer. In addition, chemotherapy-induced peripheral neuropathy (CIPN) may persist long after termination of chemotherapy. Thus, there is a need for treatments to ameliorate pain during the course of an effective cancer treatment regimen. Because the mechanism underlying CIPN has yet to be fully characterized, there is a current lack of effective treatments for CIPN. Preclinical studies in CIPN rodent models have suggested a number of potential neuropathological mechanisms, which could serve as platforms for the development of novel therapeutics. Although a number of potential analgesic therapies have demonstrated robust efficacy in preclinical studies, rigorous clinical testing has yet to fully validate the preclinical findings. The lack of congruence between preclinical and clinical findings could be in part due to the phylogenetic distance between the main model species and humans. Thus, a CIPN model in nonhuman primates could serve to bridge the translational gap between laboratory findings in small animals and clinical utility. The current review points out the short comings of current CIPN rodent models and suggests the use of large animals, such as the nonhuman primate, to narrow the translational gap between preclinical and clinical findings and the discovery of novel therapeutics.

A Phantom Pig Abdomen as an Alternative for Testing Robotic Surgical Systems: Our Experience

The use of animals for testing and validating new medical devices and surgical techniques has raised ethical issues for a long time. Following the introduction of the Three Rs principle, significant efforts have been made to achieve a reduction in the numbers of animals used in testing. Nevertheless, the number of large animals used for testing purposes is still too high. This article describes a potential alternative to the use of large animals in the early phase of the development of surgical equipment -- a high-definition phantom pig abdomen. The phantom pig abdomen was developed from computed tomography scans by using affordable materials, and it was used with two different robotic platforms. It permitted the testing of minimally-invasive robotic pancreatic enucleation, with or without intraoperative ultrasound guidance. The phantom pig abdomen has proven to be a realistic tool, with the potential to reduce the cost and time-frame of the experiments.

Attitude of Brazilian veterinarians in the recognition and treatment of pain in horses and cattle

ObjectiveThe objective of this study was to assess the use of analgesics, describe the attitudes of Brazilian veterinarians towards pain relief in horses and cattle and evaluate the differences due to gender, year of graduation and type of practice. Study designProspective survey. MethodsQuestionnaires were sent to 1000 large animal veterinarians by mail, internet and delivered in person during national meetings. The survey investigated the attitudes of Brazilian veterinarians to the recognition and treatment of pain in large animals and consisted of sections asking about demographic data, use of analgesic drugs, attitudes to pain relief and to the assessment of pain. Descriptive statistics were used to analyze frequencies. Simple post hoc comparisons were performed using the chi‐square test. ResultsEight hundred questionnaires were collected, but 87 were discarded because they were incomplete or blank. The opioid of choice for use in large animals was butorphanol (43.4%) followed by tramadol (39%). Flunixin (83.2%) and ketoprofen (67.6%) were the most frequently used NSAIDs by Brazilian veterinarians. Respondents indicated that horses received preoperative analgesics for laparotomy more frequently (72.9%) than cattle (58.5%). The most frequently administered preoperative drugs for laparotomy in horses were flunixin (38.4%) and xylazine (23.6%), whereas the preoperative drugs for the same surgical procedure in cattle were xylazine (31.8%) and the local administration of lidocaine (48%). Fracture repair was considered the most painful surgical procedure for both species. Most veterinarians (84.1%) believed that their knowledge in this area was not adequate. Conclusions and clinical relevanceAlthough these Brazilian veterinarians thought that their knowledge on recognition and treatment of pain was not adequate, the use of analgesic in large animals was similar in Brazil to that reported in other countries.

Surgical Technique for Spinal Cord Delivery of Therapies: Demonstration of Procedure in Gottingen Minipigs

This is a compact visual description of a combination of surgical technique and device for the delivery of (gene and cell) therapies into the spinal cord. While the technique is demonstrated in the animal, the procedure is FDA-approved and currently being used for stem cell transplantation into the spinal cords of patients with ALS. While the FDA has recognized proof-of-principle data on therapeutic efficacy in highly characterized rodent models, the use of large animals is considered critical for validating the combination of a surgical procedure, a device, and the safety of a final therapy for human use. The size, anatomy, and general vulnerability of the spine and spinal cord of the swine are recognized to better model the human. Moreover, the surgical process of exposing and manipulating the spinal cord as well as closing the wound in the pig is virtually indistinguishable from the human. We believe that the healthy pig model represents a critical first step in the study of procedural safety.

Surgical Technique for Spinal Cord Delivery of Therapies: Demonstration of Procedure in Gottingen Minipigs

This is a compact visual description of a combination of surgical technique and device for the delivery of (gene and cell) therapies into the spinal cord. While the technique is demonstrated in the animal, the procedure is FDA-approved and currently being used for stem cell transplantation into the spinal cords of patients with ALS. While the FDA has recognized proof-of-principle data on therapeutic efficacy in highly characterized rodent models, the use of large animals is considered critical for validating the combination of a surgical procedure, a device, and the safety of a final therapy for human use. The size, anatomy, and general vulnerability of the spine and spinal cord of the swine are recognized to better model the human. Moreover, the surgical process of exposing and manipulating the spinal cord as well as closing the wound in the pig is virtually indistinguishable from the human. We believe that the healthy pig model represents a critical first step in the study of procedural safety.

IL-12 based gene therapy in veterinary medicine

The use of large animals as an experimental model for novel treatment techniques has many advantages over the use of laboratory animals, so veterinary medicine is becoming an increasingly important translational bridge between preclinical studies and human medicine. The results of preclinical studies show that gene therapy with therapeutic gene encoding interleukin-12 (IL-12) displays pronounced antitumor effects in various tumor models. A number of different studies employing this therapeutic plasmid, delivered by either viral or non-viral methods, have also been undertaken in veterinary oncology. In cats, adenoviral delivery into soft tissue sarcomas has been employed. In horses, naked plasmid DNA has been delivered by direct intratumoral injection into nodules of metastatic melanoma. In dogs, various types of tumors have been treated with either local or systemic IL-12 electrogene therapy. The results of these studies show that IL-12 based gene therapy elicits a good antitumor effect on spontaneously occurring tumors in large animals, while being safe and well tolerated by the animals. Hopefully, such results will lead to further investigation of this therapy in veterinary medicine and successful translation into human clinical trials.

Use of Large Animals to Assess Human Safety of Heart Valves and Other Medical Devices

ABSTRACTThe use of large animals are critical for the evaluation of safety of human valves and other medical devices. Preclinical assessment includes many factors oriented at maximizing safety and performance. The final step of the device approval for patient's use goes through an exhaustive regulatory process to prevent clinical complications or any other close or distant injuries.The development of reproducible animal models, the better understanding of the implantation of medical devices and the follow through with specific clinical trials will complete the significant steps required for the evaluation of safety of human heart valves and other medical devices.

In vitro validation of a Pitot‐based flow meter for the measurement of respiratory volume and flow in large animal anaesthesia

ObjectiveTo remodel and validate commercially available monitors and their Pitot tube‐based flow sensors for use in large animals, using in vitro techniques. Study designProspective, in vitro experiment. MethodsBoth the original and the remodelled sensor were studied with a reference flow generator. Measurements were taken of the static flow‐pressure relationship and linearity of the flow signal. Sensor airway resistance was calculated. Following recalibration of the host monitor, volumes ranging from 1 to 7 L were generated by a calibration syringe, and bias and precision of spirometric volume was determined. Where manual recalibration was not available, a conversion factor for volume measurement was determined. The influence of gas composition mixture and peak flow on the conversion factor was studied. ResultsBoth the original and the remodelled sensor showed similar static flow–pressure relationships and linearity of the flow signal. Mean bias (%) of displayed values compared with the reference volume of 3, 5 and 7 L varied between −0.4% and +2.4%, and this was significantly smaller than that for 1 L (4.8% to +5.0%). Conversion factors for 3, 5 and 7 L were very similar (mean 6.00 ± 0.2, range 5.91–6.06) and were not significantly influenced by the gas mixture used. Increasing peak flow caused a small decrease in the conversion factor. Volume measurement error and conversion factors for inspiration and expiration were close to identity. ConclusionThe combination of the host monitor with the remodelled flow sensor allowed accurate in vitro measurement of flows and volumes in a range expected during large animal anaesthesia. Clinical relevanceThis combination has potential as a reliable spirometric monitor for use during large animal anaesthesia.

From the editors

From the editors