Victoria Hampshire

Using the facial grimace scale to evaluate rabbit wellness in post-procedural monitoring

Rabbits are commonly used in biomedical research and might undergo potentially painful procedures during the course of a study. This column discusses the rabbit facial grimace scale as a tool for monitoring post-procedural pain and explains how it can be incorporated into a worksheet for evaluating rabbit wellness.

The role of the veterinary staff in mouse breeding colony management

The rapid increase in the production and use of transgenic mice has been a boon for biomedical research and a challenge for the animal care and use programs responsible for providing housing and medical care to these animals. The authors suggest ways in which the veterinary staff can successfully organize and manage transgenic mouse breeding programs to reduce uncontrolled breeding and the problems associated with it.

CRISPR/Cas9 and the future of clinical research

CRISPR/Cas9 also has the potential to transform the use of animal models in safety trials. Many devices, drugs and biopharmaceutical agents that are intended for human biomedical applications must first be shown to be safe and effective; this is often accomplished using animal models. Generally, researchers subsequently characterize the effectiveness and endpoint of a treatment later during clinical trials with humans. An exception, known as the ‘Animal Rule’, permits researchers to study the effectiveness of a treatment using primarily animals because the treated condition is serious or life threatening, and it would be unethical to induce the condition for study in humans6. Using CRISPR/Cas9, researchers might be able to develop new animal models that more closely emulate human pathology, in order to better determine the safety and effectiveness of new treatments before they are deployed in humans.

Basic research support for shared magnetic resonance imaging resources.

Procedures that enable the collection of longitudinal physiologic and anatomic information can contribute to the reduction and refinement of animal use. Scientists are increasingly turning to noninvasive magnetic resonance imaging (MRI) to obtain such information from animal research subjects. As they make this important investment, research support veterinarians are often tasked with ensuring the proper care and use of laboratory animal research subjects. A basic understanding of MRI equipment, personnel practices, safety, and monitoring of animals and their recoveries is key to implementing a centralized animal MRI facility.

Primate wellness exams.

Non-human primates (NHPs) continue to serve unique animal research needs. Decades of biomedical research have focused on humane restraint and social and environmental enrichment programs necessary to support NHPs, but the housing and care of NHPs remain challenging because of the occupational safety hazards faced by employees who are tasked with the care and use of these valuable animals. Key to obtaining reliable results, providing humane care and ensuring occupational safety when working with NHPs is a sound annual wellness examination. Clinical techniques in veterinary medicine have improved in parallel with efforts to improve psychological well-being, so the timing is opportune to update clinical techniques inside the primate facility.

Simple blood typing and cross matching techniques in swine.

the techniques described below. On the day of procedure, the pigs were immobilized in their housing cages with intramuscular Ketamine (20 mg/kg), and then transported to the surgical suite. They were lifted onto surgical table and then masked with 5% isoflurane until endotracheal intubation was possible. The pigs were given butorphanol (0.2 mg/kg) intramuscularly as analgesia and maintained on a mixture of isoflurane and oxygen for the remainder of the procedure. Using the Seldinger technique, our group placed central venous and arterial lines for monitoring, blood sampling and treatment in all of our animals, making blood readily accessible for various sampling. Initial blood samples were drawn to establish blood type during baseline lab and vital signs acquisition. A blood sample was drawn, with a 5 ml initial sample discarded. We used a 9F introducer catheter (Cordis Corp, Fremont, CA) to draw the blood from the subject and the catheter was flushed with saline. The blood drawn was then transferred into a vacutainer with EDTA (BD, East Rutherford, NJ) anticoagulant and inverted carefully by hand to adequately mix the blood with the anticoagulant. It’s important to do this carefully in order not to cause hemolysis. The blood was assessed for clots after resting for approximately two minutes. the anti-A antibodies in the plasma may react with the “A” substance in the donor pig plasma to cause DIC3. It is critical that investigators have prior knowledge of these potential problems to avoid unnecessary loss of life or data. There is a paucity of literature on blood typing and cross matching in swine2, and no previous reports adequately detail these methods. To be useful, blood typing in swine should not require excessive time, incubation, specialized equipment, or specialized reagents. Additionally, there is value in universal adoption of standardized methods involving any transfusion of swine blood product. For these reasons, our Combat Trauma Research Group laboratory established standardized methods to blood type and cross match that can be used across swine models that require blood transfusions, thereby reducing complications, subject loss, and mirroring procedures routinely performed when conducting human blood transfusions. We use two methods for the forward blood typing in our swine: The EldonCard® (Eldon Biologicals, Gentofte, Denmark) and the standard saline agglutination (SSA) method. Both methods are dependent on antibodies that cause RBC agglutination, just like in human blood typing, and can be used for reverse typing, where the blood type is already known but confirmation is needed. Both methods can use fresh or refrigerated whole blood, requires minimal and easily available supplies, and have a simple quick process for results that are easily read. The EldonCard and the SSA methods described below can be completed individually or in combination with one another to cross-check and validate results.

Using the SOAP format and a minimum database with clinical chemistry

relatively similar values for key physiological parameters. Programs of veterinary care are encouraged to carefully consider the methodology that was used to establish laboratory normal values5. Neither the Guide for the Care and Use of Laboratory Animals nor AAALAC specify how veterinarians should determine a diagnosis; they only assert that attending veterinary staff should be familiar with the study species, well-versed in the diagnostic techniques needed for that species, and able to determine what could go awry, based on the pertinent procedures and interventions. They also require that veterinarians have the ability to weigh-in on important details of the protocol and to ensure that a protocol allocates appropriate resources to provide suitable care for the animals. This is especially important because insufficient planning and inadequate resources are common reasons why a timely diagnosis fails to take place. Sometimes rapid cageside monitoring equipment is not accessible or in disuse, or a monitoring time point that should take place after-hours has not been implemented. In such cases an important diagnostic value could manifest but go undetected, resulting in a failure to diagnose conditions and implement therapy or a corrective physiologic action within an appropriate timeframe. The typical veterinary program of care has access to a clinical chemistry lab for routine assessment and diagnostic purposes, but this does not guarantee a timely response to clinical problems. Most programs use the services of a contract lab that picks up physiological samples late in the day and ships them, overnight, to an outside laboratory facility. The results are then faxed or provided electronically to the program on the next day. This might be sufficient for protocols in which minor perturbations in physiologic values are expected, minimum database can describe a variety of physiological attributes and generally includes values for physiological and hematological parameters such as packed blood cells (serum hematocrit), total solids, blood glucose and serum electrolytes. The equipment needed to analyze blood samples and quantify these parameters is inexpensive by the standards of laboratory animal science (Table 1) and does not require complex calibration. Where diagnosis and clinical response are concerned, recent literature emphasizes the need to monitor and treat pain; however, dehydration, anemia, electrolyte deficiencies and hypoglycemia are also common conditions that, when detected early in their development, can be quickly corrected to optimize humane and experimental outcomes.

Improving survival of animal models with implanted artificial heart valves

adverse outcomes to the US Food and Drug Administration such that any potential risk to safety can be accounted for in the test device, the procedure or some other confounder. Often this accounting requires a timely and accurate diagnosis of cardiac events or any deviant hematology, acid-base disturbances and pulmonary, renal or neurologic findings. A strong level of proficiency in clinical and cardiac techniques and physiology is needed to diagnose such errant conditions and whether they are directly attributable to the device or procedure. Clinicians also need a strong understanding and familiarity with many classes of drugs and constant rate infusions that are not commonplace at traditional laboratory animal facilities. Boardcertified veterinary cardiologists or related echocardiographic experts are suggested are required to navigate these difficulties, and clinical practices can be improved with key personnel, equipment and methods to optimize the successful creation of animal models, ultimately reducing waste and animal distress.

Altered states part 1: ancillary refinements in managing perioperative distress with canines

The biomedical research community has made great progress over the last 30 years in improving strategies to detect and manage pain. Managing experimental and procedural anxiety is a more challenging task, and it depends on the continued practice of distinguishing normal and abnormal states of physiology and behavior in animal subjects. Common approaches for managing pain and distress can be optimized by implementing a plan to also monitor and manage anxiety and dysphoria.

Altered states part 2: addressing nausea in canine research subjects

Nausea and emesis can occur for multiple reasons. While research staff can readily empathize with this type of discomfort, proper assessment and treatment can be challenging. In order to provide optimal care for canine research subjects, it is critical that institutions develop a treatment plan and take preemptive measures to control nausea and emesis when they occur.