Session Four: Restraint and Handling of Animals in the Field to Include the Use of Compounds for Capture and Handling

Moderator: Bonnie V. Beaver, Texas A&M University

Bonnie V. Beaver, a veterinarian and professor of biology at Texas A&M University, opened Session Four, which focused on considerations related to the restraint and handling of animals in the field, including the use of drugs to facilitate safe handling, as well as controlling pain during certain invasive procedures, and related implications for the U.S. Department of Agriculture (USDA) pain categorization of animal activities. Speakers discussed the challenges of using non-pharmaceutical grade drugs and controlled substances during various types of wildlife research and the challenges with using them in the field; the release of animals that have been exposed to chemical compounds in the field (including potential consumption by humans or other animals); the role of wildlife agencies, wildlife veterinarians, and relevant regulations in this space; and the challenges and options for non-veterinarians, particularly for the use of controlled substance and extra-label drug use (ELDU, as it is commonly known among wildlife professionals).

EXTRA-LABEL DRUG USE IN FREE-RANGING WILDLIFE: RULES, REGULATIONS, AND REALITY

Lisa A. Tell, veterinarian and faculty member with the University of California (UC), Davis, School of Veterinary Medicine was clinically trained at the Smithsonian National Zoological Park and worked with wildlife for her entire career. She currently serves as the western regional director for the Food Animal Residue Avoidance Depletion Program. Tell explained that Food Animal Residue Avoidance Depletion Program is a USDA-sponsored program that provides veterinarians with withdrawal interval recommendations when drugs are administered to food animals in an extra-label manner and the Food Animal Residue Avoidance Databank (FARAD) 2022 is a component of the program. This program is a collaboration between the School of Veterinary Medicine at UC Davis and several other U.S. Veterinary Colleges, including those at Kansas State University, North Carolina State University, the University of Florida, and the Virginia-Maryland College of Veterinary Medicine at Virginia Tech. Tell presented information for on-label drug use versus ELDU, including rules and regulations for animals that may be consumed by humans; requirements for legal ELDU; challenges with trying to establish withdrawal intervals following ELDU in wildlife; and provided some available resources.

Tell explained, with any drug use there is the potential for drug residues to be present in animal tissues and products. She said that technically a drug residue is the parent or the metabolite that can accumulate in edible tissues or products (e.g., eggs, honey) from a treated animal. Tell said that as a measure to protect animal and human health, the U.S. Food and Drug Administration (FDA) approval processes requires the drug be used in accordance with an FDA-approved product label, regarding species, dose and routes of administration, frequency of administration, treatment duration, and indications for use. On-label drug use also applies to limitations including drug withdrawal time, Tell added. She explained that to avoid unsafe residue concentrations, the FDA-approved withdrawal time needs to be observed. This is the time that it takes for the drug to metabolize to a safe concentration in live animals before edible animal tissues and products are safe for human consumption.

Tell said that the withdrawal time is designed to protect humans, so they do not consume drug residues at concentrations that could have negative effects on them. A classic example of a residue toxicity case was the illegal use of Clenbuterol to increase weight gain in calves. That resulted in numerous people developing heart palpitations, abnormally fast heart rates, and muscle tremors after eating veal liver. Other examples included penicillin allergic reactions and chloramphenicol-induced bone marrow

suppression. In contrast to FDA-approved, on-label withdrawal time, Tell described the term withdrawal intervals, which is an estimated withdrawal period when drugs are administered in an extra-label manner. Tell added, it is important to know both ELDU withdrawal intervals and FDA-approved withdrawal times for safe consumption and to prevent direct impacts (quick reactions [i.e., toxicity, allergic reactions, and bone marrow suppression]) and indirect impacts (long-term reactions [i.e., carcinogenicity, mutagenicity, and reproductive disorders]).

Tell shared the label information for AnaSed® Injection (xylazine) 100 mg/ml, which listed the species, dose information, indications, and limitations (e.g., withdrawal time). She provided the FDA-approved formulations for the wildlife species and the number of approved products, as shown in Table 4-1. Tell said that there are fewer unique medications because the active ingredients in several different products are the same. For example, Table 4-1 shows the information Tell retrieved from the FARAD VetGRAM¹2022, where there are a total of 14 FDA-approved products listed for cervids but there are only 6 unique FDA-approved formulations, more commonly referred to as active ingredients. Tell showed the FDA-approved meat withdrawal time (MWT) for deer and elk: AnaSed® and Chanazine® (xylazine), which stated not to use less than 15 days before or during the deer and elk hunting season. The Antagonil® (yohimbine) product label read do not use less than 30 days before or during the hunting season. Tell said, however, if xylazine was administered during the hunting season that is considered ELDU and the on-label withdrawal time of 15 days would not apply and would need to be extended. For ivermectin use in reindeer, Tell said that there is a 56-day withdrawal time. Tell said that this means that the animal should not be harvested (hunted) for 56 days following drug administration to avoid unsafe drug residue concentrations during human consumption.

In contrast to FDA-approved on-label drug use, Tell said that ELDU differs for animal species and class, administration route, dose, dosing frequency and interval, duration, and treatment indications and limitations (e.g., injection volume and site, withdrawal time). ELDU was legalized by the Animal Medicinal Drug Use Clarification Act (AMDUCA) (FDA 1994). Tell said that over the 10-year period of 2012–2022, ELDU requests to FARAD for ELDU in wildlife were only 4% of all submissions. Tell found that low number surprising, given the small number of FDA-approved formulations for wildlife. She hypothesized that it could be because veterinarians do not know about the FARAD services, or that possibly ELDU is occurring without veterinary oversight. Of the wildlife submissions to FARAD, about half were for waterfowl, followed by cervids, game birds, and wild boar; and the remainder were small percentages for different types of animals. Tell said that the top 10 requests to FARAD drug use in an extra-label manner for wildlife during this same 10-year period were:

• Meloxicam (pain medication)
• Amoxicillin trihydrate-clavulanate (antibiotic)
• Enrofloxacin (antibiotic); ELDU of fluoroquinolones in food animals is prohibited so FARAD does not provide a withdrawal interval for Enrofloxacin
• Oxytetracycline (antibiotic)
• Carprofen (pain medication)
• Fenbendazole (antiparasitic)
• Xylazine (anesthetic agent/sedative/tranquilizer)
• Butorphanol (pain medication)
• Tiletamine hydrochloride-zolazepam HCL (anesthetic agent)
• Clindamycin hydrochloride (antibiotic)

___________________

¹ VetGRAM is the Veterinarian’s Guide to Residue Avoidance Management, an informational resource for approved uses, restrictions, and required withdrawal times for drugs approved in food animal species. Information is exclusively for labeled use(s) of FDA-approved food animal drugs in the United States.

TABLE 4-1 U.S. Food and Drug Administration (FDA)-Approved Formulations for Wildlife Species and Number of Approved Products

SOURCE: Adapted from Tell presentation, taken from FARAD VetGRAM 2022.

There are several stipulations for legal extra-label administration of drugs. If an FDA-approved product exists and fits the medication need, then that product has to be used, before going extra-label, unless there is a compelling reason for why that product cannot be used. If there is not an FDA-approved product, extra-label use is necessary, then there has to be a veterinarian–client–patient relationship (VCPR) with a licensed veterinarian to supervise the use of the drug in an extra-label manner.

With wildlife cases, the traditional VCPR may not be possible, but FARAD does consider licensed veterinary supervision of biologists, game wardens, or others administering medications in an extra-label manner to fulfill that requirement. In addition, ELDU is only permitted for FDA-approved human or veterinary prescription or over-the-counter (OTC) products. Not all OTC medications are approved by the FDA and ELDU of unapproved drugs are not allowed.

Tell said that the AMDUCA stipulates that FDA-approved drug use in an extra-label manner only applies to therapeutic uses, not for production purposes, convenience, or cost-saving measures. “Therapeutic” refers to situations where suffering or death may occur if treatment is not administered. For

wildlife, FARAD would consider several situations to fit into this category of “death may occur if treatment is not administered” (i.e., behavioral management), Tell said. Another requirement is the identification of treated animals. For free-ranging wildlife, it is especially important to tag or mark animals receiving ELDU, regardless of whether they are being treated before, during, or after the hunting season, given all of the potential times and ways that a carcass could enter the human food chain. These tagging and marking techniques will be discussed in Session Four, Talk 2 of the proceedings.

The AMDUCA also requires that a substantially extended withdrawal interval be established based on scientific data. It stipulates that assurances have to make certain that illegal drug residues will not be present in order to protect human health. In addition to human health, it is also important to be mindful of the potential for consumption and secondary poisoning of other animals. Due diligence to thoroughly document drug administration is essential, she stated.

Tell asked How would a substantially extended withdrawal interval be estimated based on scientific data? FARAD’s most common method is to use published pharmacokinetic studies, she said, but it is challenging with wildlife because of the limited numbers of studies. Of the 16,000 studies in FARAD, only about 500 are on wildlife. Therefore, researchers have to extrapolate from studies conducted on domestic species. Another option is to extrapolate from an FDA-approved withdrawal time for a domestic food animal product or extrapolate from foreign drug approvals. If available, pharmacokinetic models can also be used to establish a withdrawal interval, Tell explained, but typically these models are for domestic species. If there is no sound scientific data or if a reasonable extrapolation cannot be made, FARAD will not be able to provide a withdrawal interval recommendation.

For free-ranging wildlife, drug delivery systems could impact drug depletion rates, Tell said. Often a remote delivery system is used that could affect drug absorption and elimination, which has to be considered when estimating withdrawal intervals. Another challenge with establishing ELDU withdrawal intervals for wildlife is that extrapolation from pharmacokinetic data for other species is often necessary. However, the drug metabolism might be different, the primary (marker) residue may be different, and the percentages of the parent drug and drug metabolites might not be the same as in other species. Given these unknowns, ELDU withdrawal interval recommendations for wildlife are often conservative.

Tell pointed out situations when withdrawal intervals cannot be provided and animals should not be allowed to enter the human food chain; for example, when a prohibited drug for any use is administered. One prohibition to highlight is ELDU use of FDA Indexed Drugs, which cannot be legally used for animals that have the potential for entering the human food chain. The required labeling language is “This product is not to be used in animals intended for use as food for humans or food-producing animals.”

As the final subject, Tell explained compounding, which by definition, is the combining, mixing, or altering of ingredients to create a medication tailored to the needs of an individual patient. Compounding is permitted as long as ELDU stipulations are followed, and the compound is made from an FDA-approved animal or human drug. Compounding from bulk powder is not allowed. Additionally, compounding from a human drug for a food-producing animal is not allowed if there is an animal drug that can be used instead. Compounding for wildlife presents another challenge because there are limited pharmacokinetic studies for compounded formulations and without these data a withdrawal interval really cannot be estimated. Finally, compounded preparations are considered unapproved new animal drugs because they have not undergone the safety and efficacy testing.

Resources for on-label drug use and ELDU are available on the FARAD website,² which provides an online interface where veterinarians can request withdrawal interval advice. The website also provides citations for the information along with a digest dedicated to ELDU and wildlife that FARAD publishes.

___________________

² See http://www.farad.org.

In closing, Tell recapped key points about ELDU in wildlife:

1. There is such a small number of FDA-approved medications for use in wildlife species that the administration of drugs in an extra-label manner is common. However, it is important to remember that only FDA-approved medications, both prescription and OTC drugs can be used; but FDA Indexed Drugs are prohibited.
2. The use of drugs in an extra-label manner requires veterinary oversight and, to be legal, non-veterinarians administering medications to wildlife need to be guided by veterinarians.
3. Ultimately, the goal is to protect human health; therefore, animals that are administered medications that require withdrawal intervals should be tagged or marked following ELDU to minimize the hunting of animals before drugs are sufficiently metabolized and thereby avoid the tainting of human food products with unsafe levels of drug residue.

THE USE OF PHARMACEUTICAL AGENTS IN CARNIVORES, BEARS, AND UNGULATES WITHIN A WILDLIFE MANAGEMENT AGENCY (REALITY AND CHALLENGES)

Mark Drew is a wildlife veterinarian with the Wildlife Health Services, LLC, and previously worked with the Idaho Department of Fish and Game (IDFG) for 21 years. Drew discussed the use of pharmaceutical agents in wildlife by a wildlife management agency, with a specific reference to the Idaho state wildlife management agency. Not all veterinarians have the same interest, expertise, or experience as Drew and information could also be sought from other highly qualified subject-matter experts.

Drew said that it is important to understand the role of wildlife management agencies, which is the sustainable management and harvest of wildlife in natural environments. In other words, the agency is charged with maintaining harvestable surplus of wildlife and fish for hunting, fishing, and trapping. In North America, wildlife is broadly classified or considered to be those species that are free-living and not domestic or are feral. Wildlife includes animals in all taxa, including amphibians, reptiles, birds, and mammals.

In general, wildlife is managed by state, federal, or tribal governments, Drew explained. The management of wildlife is a partnership between public land management agencies and private landowners because many wildlife species occur on both, he added. Because the most important component of the existence of wildlife is appropriate habitat, the management of various ecosystems and habitat types is central to agency work. The North American Model of Wildlife Conservation is how wildlife is managed in the United States and consists of two basic principles: (1) that fish and wildlife belong to the people, and (2) that wildlife populations need to be managed in a way that their populations will be sustained forever. These principles form the foundation of wildlife management in the IDFG, as well as elsewhere in state wildlife management agencies in the United States.

Currently, the IDFG has approximately 569 employees spread across seven bureaus: fisheries, wildlife, enforcement, communications, engineering, information systems, and administration. There are approximately 100 individuals working in the Wildlife Bureau across the state as biologists, technicians, or researchers, and approximately 100 game wardens in the Enforcement Bureau.

To accomplish various management and research objectives, Drew said, many wildlife species are captured and handled by IDFG personnel, including biologists, game wardens, technicians, and veterinarians. The restraint and handling of wildlife in the field by veterinarians and biologists often starts with chemical immobilization agents, but also includes the administration of antibiotics, anti-inflammatory agents, vaccines, and anthelmintics. For the past 10 years, the IDFG has captured and handled around 1,100 animals per year of many species, some using physical restraint methods and about half of them using a variety of chemical immobilization drugs.

Drew said for chemical immobilization or restraint, both controlled and non-controlled substances are used. Drew commented that Tell already provided information about FDA-approved drugs and their use under the AMDUCA, including drug use in an extra-label manner. Some of the drugs used by the IDFG are FDA-approved drugs; some are Drug Enforcement Administration (DEA)-controlled

(scheduled) substances; some are commercially available drugs; and some are compounded by veterinary compounding pharmacists. Most are used in an extra-label manner, although some drugs are actually approved for the species in which they are administered in the state. The drugs that are used in Idaho include:

• Narcotics (Etorphine, Butorphanol, Nalbuphine)
• Neuroleptic tranquilizers (Acepromazine, Azaperone, Haloperidol, Diazepam, Midazolam)
• Sedatives (Xylazine, Detomidine, Medetomidine)
• Cyclohexamines (Ketamine, Tiletamine in the form of Telazol®)
• Antagonists or reversal agents (Naltrexone, Yohimbine, Tolazoline, Atipamezole, Flumazenil)
• Index-approved narcotics from under an Investigational New Animal Drug (INAD) with the supplier of the drug (Thiafentanil)

Drew said that due to the constraints of dart size, many of the drugs used for chemical immobilization are compounded to a higher concentration than commercially available, to accommodate a smaller dart size. The products that they use are produced by compounding veterinary pharmacists under strict compounding manufacturing practices and existing compounding rules. Drugs are also mixed to a higher concentration than what is standard on the label, or in non-standard combinations like xylazine and Telazol®. Many drug combinations are available as pre-mixed, and some are super concentrations of drugs.³ This greatly enhances the ability of people working with wildlife to put enough drug in a small enough volume to minimize impact trauma and ensure complete absorption of the drug in a prompt manner, Drew said.

Wildlife veterinarians are the primary drug conduit within wildlife management agencies, Drew explained, while most administration is done by wildlife biologists with minimal direct supervision by the veterinarian. It is important to understand that Drew was the only veterinarian for the Wildlife Bureau and his practice area encompassed the entire state. It could take 8 hours or more to drive to northern Idaho and 6 hours to get to eastern Idaho. By the time he could get to the location of the moose or bear in town, the local biologists or game warden could easily have taken care of the situation. In addition, he could not be in more than one place at one time, Drew added, and there were times when multiple situations or capture events were happening at the same time in multiple areas in the state.

Drew said that because most of the administration of drugs is handled by biologists or game wardens; these individuals worked under a VCPR with him as the veterinarian. That VCPR starts with a required 16-hour class to attend every 2 years for anyone expected to use or wanting to use controlled substances or other drugs for required chemical immobilization. As a veterinarian, Drew was active in field activities as much as possible, with direct contact with biologists and game wardens.

Drew said the VCRP also included the provisions of the AMDUCA that for any ELDU under a VCPR, all animals were required to be identified and MWTs were established for each of the drugs for each of the species for which they were used. Drew stated, to comply with the AMDUCA, the IDFG policy included the following:

___________________

³ Drug combinations are available pre-mixed like BAM (e.g., Butorphanol-Azaperone-Medetomidine); or Ketamine-Xylazine 5:1; Ketamine-Metomidine 15:1; or NalMedA (Nalbuphine-Medetomidine-Azaperone). In addition, several drugs of high concentrations are mixed independently into appropriate drug combinations like BAM2 (2× Butorphanol-Azaperone-½ Medetomidine) or BMM (Butorphanol-Midazolam-Medetomidine). Some of the concentration of drugs that are available through compounding veterinary pharmacists are super-concentrations of drugs: Xylazine (200, 300, 333 mg/ml), Ketamine (200 mg/ml), Butorphanol (30, 50 mg/ml), Midazolam (50 mg/ml), Medetomidine (10, 20, 40 mg/ml), Azaperone (50 mg/ml), Atipamezole (25 mg/ml), compared to the standard commercially available concentrations.

• As a licensed veterinarian, IDFG personnel were working under him with VCPR and IDFG personnel as the actual animal’s agent.
• The IDFG required identification of each animal receiving drugs by using an ear tag that provides information to contact the IDFG Wildlife Health Laboratory before human consumption.
• Drug use data were required to be reported to the veterinarian’s office within 48 hours after drugs were administered, including subject signalment, date, location of the animal, and drugs used and the dosage. This allowed the veterinarian to determine the MWTs established using either FARAD or published references.
• Information was then entered into a database that could be retrieved if an individual harvested a tagged animal and contacted the IDFG.
• If the animal was harvested within the MWT, the person in possession of the carcass was asked, but not required, to surrender the carcass. That allowed the IDFG to dispose of the carcass in the nearest landfill. An option was also offered for issuing a new tag for the same species and the same season if the tag was still available.

Drew said that the reason for IDFG capturing, or handling wildlife was generally to assess management actions or conduct research with the goal to release animals unharmed into their normal environment. It was not possible to prevent predation and consumption of drug residue in prey species because these animals were released back into their environment as soon as possible after handling. In Idaho, the complete suite of predators includes wolves, coyotes, mountain lions, lynx, bobcat, black bears, and grizzly bears, Drew explained. If animals died during capture or were euthanized due to injuries from capture, carcasses were retrieved, if possible, and then taken to the nearest landfill or burial pit to prevent relay situations. If retrieval was not possible, the carcass was generally left in the field. It is important to understand that the animals captured and handled by the IDFG were not directly intended to be food animals, even though they are considered game species and could be harvested for food.

Animals were handled as part of long-term management or research efforts with the goal to keep them on the landscape for many years, Drew said, not to subject them to immediate harvest. Most animals are also captured or handled outside the typical hunting season whenever possible, but there were also situations, particularly with animals in town, where they were handled and/or released during an active hunting season (IDFG 2021). Drew said that it is important to understand how hunting works in Idaho and some of the issues that hunting brings up, in general. The IDFG sets the seasons for big game, small game, and trophy species harvest but it is not a simple one season for everyone, he said. For example, for black bear, there are spring rifle, fall rifle, archery, and dog seasons. What that means is that some bear populations potentially have harvest seasons that may last many months. Similarly, for elk, there are spring green field hunts, fall rifle, archery, and muzzle loader seasons, and winter depredation hunts, which means that some elk populations are also subject to harvest for several months.

Drew said that it was important to understand that hunter success is relatively low, and the actual risk of the few individuals that are handled with chemical immobilization drugs being harvested by hunters is relatively low. For instance, if there were 500 to 1,000 animals that received chemical immobilization drugs in each capture season, in the upcoming harvest season when the population consists of potentially 50,000, 60,000, or 100,000 animals, those individual animals are rarely harvested. It is also important to understand that there is tribal harvest for the five recognized federal tribes in the state that have different bag limits and seasons than those set by the IDFG. A further complication is that in Idaho, there is a wildlife salvage rule where citizens can collect wildlife that die naturally, including those hit by cars or other vehicles, for consumption or other uses. There are also considerations such as poaching, Drew added.

Another complication or issue of concern is that in Idaho as well as in other states, Drew said, wildlife moves, sometimes long distances. Winter and summer migrations may be dozens to hundreds of miles, and some animal species follow in plant phenology in elevation, sometimes for dozens or 50 or

more miles. Wildlife cross state and international borders, sometimes without even the benefit of a health certificate. Drew pointed out that the IDFG is not the only agency that handles wildlife in Idaho. Although the IDFG is the primary agency, there are many other entities that handle animals. Some of the above activities are known to the IDFG through a permitting system; others are not. For example:

• Five Native American tribes that have their own wildlife management agencies that handle animals of a variety of different species:
  • Shoshone-Bannock
  • Shoshone-Paiute
  • Coeur d’Alene
  • Kootenai
  • Nez Perce
• USDA Animal and Plant Health Inspection Service: U.S. Forest Service and U.S. Fish & Wildlife Service (FWS)
  • Predators and species that may negatively impact agriculture
• U.S. Department of the Interior (DOI): Bureau of Land Management (BLM), FWS, and National Park Service (NPS)
• U.S. Department of Commerce (DOC): National Oceanic and Atmospheric Administration (NOAA)
• Academia: university researchers and graduate students handle wildlife
• Other activities: wildlife rehabilitation involving both the capture and the release of animals that are outside the scope of the IDFG

Drew concluded by making an implicit request to the workshop participants to understand the many complexities of wildlife management, including the role of wildlife management agencies and the role of veterinarians within those agencies. They try to balance the need for appropriate treatment of wildlife with regulatory oversight, particularly for the chemical immobilization drugs. It can be challenging because there is oversight by multiple regulatory agencies, some of which is contradictory, some of which is not helpful, and some of which restricts the use of the most appropriate or effective drugs for species being handled.

Drew said that, while it may be simple to say “thou shalt not,” the risk is that the handling, treatment, and, ultimately, the welfare of free-ranging wildlife for management, research, or health would be severely compromised without access to appropriate pharmaceutical agents for the species of concern. He added that the only alternative would be euthanasia for all animals that cannot be confined or held to fulfill the MWT—or if the drugs available are not effective or appropriate for the species of concern in the first place.

A BRIEF INTRODUCTION TO THE COMPLEXITIES OF WORKING WITH MULTIPLE AGENCIES TO CONDUCT WILDLIFE ANIMAL USE ACTIVITIES

John A. Bryan II, a freelance wildlife veterinarian who has worked with academic, federal, private-sector, state, and international agencies and entities, including those of foreign governments, shared his insights on some of the challenges of wildlife fieldwork from a broad global perspective. To begin, Bryan provided an overview of the different levels of oversight of wildlife animal use activities (WAUAs) in the United States:

• State-level oversight: The specifics of oversight of free-ranging species of wildlife vary from state to state and from territory to territory. However, the general concepts undergirding state mandates are relatively consistent. State wildlife agencies, with some exceptions, hold final authority on wildlife oversight within their boundaries. For example, the principal agent,

• authority, and manager of free-ranging species in each state is that state’s recognized wildlife agency. Three of the central ways in which state wildlife agencies fulfill this role or meet these responsibilities involve wildlife management, research, and education and outreach. State wildlife agencies are primarily mandated to manage free-ranging populations of animals and they often do this via conservation, disease management, and outbreak response, permitting for legal take activities (e.g., hunting and fishing) and population management (e.g., culling and/or breeding efforts). This is not a complete list of the duties under the purview of state wildlife agencies. However, these are the main issues that often directly impact activities involving animal capture, handling, and pharmacology within the state. In the United States, for these reasons, states form the first level of bureaucracy and regulation with which any WAUA must coordinate.
• Federal-level oversight: The federal level of wildlife oversight is more focused or narrow when compared to the state level. For the most part, federal wildlife oversight concerns the conservation and protection of certain species. Typically, the management of subspecies falls into the authority of the federal agency tasked with administering a law designed to protect that species. For example, because they are covered and listed under the Endangered Species Act (ESA), final authority over the management of Florida panthers and California condors rests with the federal government, specifically the FWS and not the states of Florida and California, respectively (taxonomic exemptions versus states). This does not preclude collaboration and/or the federal agency delegating some responsibility to a state agency, but the hierarchy is relatively established. In a similar manner concerning marine mammals, such authority likewise falls to the NOAA. The NPS is in a unique position, however in that, unlike most other federal agencies the NPS does have final management and oversight authority over wildlife within its geographical jurisdiction (territorial exemptions versus states), unless the species in question is listed under the ESA.
• Academia and private sector: In general, academic institutions and private-sector entities hold no inherent authority to manage wildlife or the habitats in which they live. However, state and federal agencies can, by special arrangement, collaborate with and even delegate certain aspects of management oversight to academic and/or private-sector groups or entities. These entities can and do conduct WAUAs, yet they must collaborate with state and/or federal agencies to proceed. They cannot lawfully unilaterally engage in wildlife animal use activities. In other words, academic and private-sector entities must conduct their wildlife animal use activities in accordance and compliance with their own policies and the laws and regulations of state and federal wildlife agencies. Such permissions often come in the form of licensure and permits from the appropriate state and federal agencies.
• International arena: From an international perspective, the host countries in which the activity takes place have the ability to require full compliance with any and all of their own regulations and demonstrating that home requirements have been met may only address half or less than half of the host country requirements. Host countries can and often do require completion of their regulations, as well. Working internationally may, and often, does demand two distinct and possibly redundant comprehensive sets of qualifying dossiers to proceed with a project. All aspects of regulation can be in play for all countries involved, for example, permits, licenses, pharmacology, and others. Bureaucracy on this level can be daunting and the complexities associated with the regulatory requirements of adding wildlife drug, capture, handling, and sampling protocols can be profoundly demanding.

His overall take-home message, Bryan said, is that challenges to successfully conduct WAUAs in the field start before any “boots hit the ground.” Regardless of the purpose, state, federal, academic, private-sector, and/or international regulations and oversight concerning the use of wildlife must be followed. Achieving compliance with all necessary regulations on each of these levels is a demanding endeavor, he said. Although IACUCs and foreign animal welfare oversight bodies have direct oversight in

reviewing and approving animal use activities, approval for such activities often includes a full accounting of having obtained all required permissions and regulations at every bureaucratic level.

Moreover, horizontal differences and regulations may also pose additional challenges, Bryan added. For example, WAUAs taking place in multiple states may face different laws and regulations from state to state further compounding efforts in project continuity and protocol application. In addition, team members licensed and permitted to conduct activities in one state may not be allowed to conduct the same activities in another state. This is especially true concerning DEA regulations and veterinary licensure; and such challenges can be amplified when working with variable formulations of chemical capture protocols, endangered species, multiple locations or states, and/or work in the international arena.

Bryan presented what he said is a “nightmare scenario” regarding the possibility of a WAUA involving the repeated capture and handling of an ESA-listed species that migrates between the United States and another country, with stopovers in multiple states. Such a project would likely require:

1. On the U.S. side, permissions from the FWS, the NOAA (possibly, if the species is a marine mammal), the NPS (possibly, if applicable), and each U.S. state in which the species will be engaged—this includes capture and handling permits, drug permits, and/or veterinary licensure (if applicable), etc.
2. On the international side, similar, if not identical, permissions from those of the United States and from all other counties with which the species will be engaged.
3. Review and approval from any academic and/or private-sector IACUC with the jurisdictional claim that may demand that the team present, prior to the start of work, obtained permissions from all of the applicable aforementioned.

Challenges in conducting work concerning the use of wildlife in the field are many, Bryan said, but must be considered by wildlife animal oversight bodies when conducting assessments of WAUAs. If these challenges are not met or appreciated, the consequences can result in delayed wildlife projects, loss, and or/waste of funding; inadequate allocations of resources; and lower standards of animal welfare, among others.

CHALLENGES IN THE CAPTURE AND HANDLING OF WILD BIRDS IN RESEARCH

Jeanne M. Fair is a scientist at the Los Alamos National Laboratory who specializes in the ecology of infectious diseases and ornithology. In addition to serving as chair for the IACUC for 10 years, Fair was editor-in-chief of the third edition of the Guidelines to the Use of Wild Birds in Research (Fair et al. 2010). Her presentation focused on some of the challenges of doing research on wild birds in the field and wild birds brought into captivity. This includes working with IACUCs and biosafety professionals to better understand the limitations in fieldwork and sharing some of the most useful resources that are available to everyone.

While both basic and applied research of birds has been going on for hundreds of years, birds themselves have been facing challenges in the past several decades, Fair said, which has led to large-scale population declines and impacts on individuals. A 2019 study documented an estimated loss of 3 billion birds, or almost a 20% loss of individuals since 1970 in North America (Rosenberg 2019). Basic science and natural history studies of birds are still vital, Fair added, but researchers are now forced to investigate the different impacts to birds in conservation efforts. Young research professionals may be opting to conduct more applied research on birds to better understand how to mitigate potential impacts to populations and to species, she said.

Free-ranging wildlife is the basis for thousands of studies each year globally, and with a worldwide abundance estimate of 9,700 bird species, birds are considered one of the most diverse taxon groups. Fair said that ornithologists contribute to biology by studying the ecology of birds in the wild and identifying the key threats and ways of enhancing the survival of species. Due to the wide diversity of bird species and field situations, methods for the capture and handling of wild birds vary just as much,

Fair explained, and may also vary dramatically for a single species depending on the field conditions. Ornithologists understand the best methods to use, she said, and it is their responsibility to know the nuances of capture and handling techniques and the state of the art for reducing impacts on birds.

Fair said that there are many misconceptions about working with wild birds and birds in general that permeate society, as well as science trained professionals. One study found that 19% of a university’s science students believed that touching nestlings will result in nest abandonment; however, there are mitigations that ornithologists can make to avoid leading predators to nests, which is a different issue (Cardak 2009). This may vary depending on the species and the field situation, Fair added. Ornithologists regularly work to address misconceptions when they are working with their IACUC.

Much of the field research on wild animals seeks to investigate behavior: examples include habitat use given habitat fragmentation or loss, movement, interactions with predators or competitors, or the rates of survival and productivity. If that is the case, Fair said, then actions could go beyond what is required by the IACUC and could focus on having the least effect possible, with the goal being no affect on the subject animals during the capture, handling, marking, and tracking. Fair said that in almost all cases scientists could understand the impact they are having on the organisms they research. However, scientists also seek to be able to ask challenging questions that remain for wild birds and to not be overly hindered to the point of not being able to ask the important questions.

Fair explained that the role of the IACUC is critical for ensuring that the research on animals is conducted humanely, while reviewing that the science is designed to answer the questions at hand. This includes the research protocol review and the semi-annual program reviews and animal facility reviews. If the institution is accredited with AAALAC it will conduct reviews onsite every 3 years. The IACUC is responsible for the oversight of the animal care and use program and its components as described in the U.S. Public Health Service (PHS) Policy on Humane Care and Use of Laboratory Animals and the Guide for the Care and Use of Laboratory Animals (the Guide) (NIH OLAW 2002; NRC 2011). There are many resources for researchers and IACUCs for laboratory animal research, including the Guide, which is recognized as the definitive source by IACUCs, as well as others (NRC 2011).

The role of the IACUC is detailed in legislation and binding rules, which are in turn inspired by the three Rs—the principles of replacement, reduction, and refinement. Fair said that it has been suggested that there is even a fourth R, reject, which allows the IACUC to refuse permission for a project that does not promise sufficient benefit to offset the pain and distress likely to be caused by the proposed research. Fair indicated that replacement would mean a substitution for conscious living higher animals; refinement would mean any decrease in the incidence or severity of inhumane procedures applied to those animals that still have to be used; and reduction would mean the reduction in the number of animals used to obtain the information of a given amount and precision. When it comes to fieldwork with wild birds, the sample size is important to be able to adequately test the proposed hypotheses, Fair said. Depending on the focal species and the field situation, sample sizes may be the hardest to improve upon. Unlike in laboratory settings, there is considerably more variability in the field, which is important to take into consideration when designing field investigations on wild birds.

Most of the refinements in ornithology have come from the improvement of techniques and technologies used to study wildlife; for example, using remote technologies that may be more useful than capturing birds, Fair said. However, it is important to understand the impacts that new technologies may have on bird behavior. For example, in the case of drones, they may be extremely disruptive to nesting birds and have been the cause of having large-scale nest abandonment of thousands of nesting seabirds at once. Often, the refinement of studies comes while conducting the study itself, she added. It is important for ornithologists to continue to publish studies on the impacts of methods on individuals and populations, especially if negative impacts are documented.

While not as common in wild bird research, replacement does occur in investigations and it may be the only option when it comes to threatened and endangered species, Fair asserted. Much of the research on wild birds is specific for understanding the species and/or local situations in the field and requires data collection about that population or field site.

Fair said that all investigators, from principal investigators (PIs) to all students involved in research projects, should be adequately trained for all the aspects of field research and the capture and handling of birds. Additional training for biosafety and field safety may be required depending on the project, she added. Training of animal handling for wildlife projects is most often completed by senior researchers and not through IACUC trainings, although both may be mandatory for researchers. It is important for IACUCs to understand the value of knowledge transfer from senior researchers to younger research professionals and students. It is helpful for members of IACUCs to learn about and participate in IACUC wildlife 101-type training events and to go into the field with researchers, not with the intent of oversight, but for learning about wildlife research and the constraints and limitations that may be present in the field. Additional insights can allow IACUCs to become adaptable to understanding method changes and in the evaluation of research protocols.

There are numerous resources for the different taxon groups for wildlife that can be used by both researchers and IACUCs. One of the most comprehensive resources for wild birds, Fair said, is the Guidelines to the Use of Wild Birds in Research originally published in 1975 by the American Ornithological Union, as it was known at the time (Fair et al. 2010). It was first called the American Ornithologist Union ad hoc Committee on the Scientific and Educational Use of Wild Birds. Fair said that, in 2010, ornithological experts were invited to edit the guidelines. The team of ornithologists completed thorough literature reviews of all peer-reviewed papers and the gray literature that documented the best described methods for the capture and handling of wild birds in the field and in captivity. The current and third edition of the guidelines are peer-reviewed and contain science-based discussions of the best practices for most aspects of research with wild birds in the wild and in captivity. It also details research that has documented the impacts of studies and scientists on birds. The book’s eight chapters cover permits, investigator impacts, scientific collecting, marking, transporting, housing and captive breeding, minor manipulative procedures, and major manipulative techniques. Each chapter outlines the best practices and known issues for most techniques used to study wild birds. In 2021, the Ornithological Council updated all of the references of literature on the methods and impacts to birds that have been added to each chapter, including the impacts of drones.

All researchers in the United States must obtain different types of permits to be compliant for research on birds in the wild and wild birds brought into captivity, Fair said. These include Migratory Bird Treaty Act permits, Bird Banding permits, and federal and state scientific collecting permits. Researchers may need permits to work on public lands such as national forests or national parks and state lands and have approval for working on private and Native American tribal lands. There also may be a need for ESA permits, Convention on International Trade in Endangered Species of Wild Fauna and Flora permits, and/or certain Certificates of Scientific Exchange.

While predicting every scenario that may occur in field research is impossible, Fair said that the field collection of biological samples does contain risk and safety concerns in general in a field setting. The practice of biosafety is traditionally limited to the laboratory setting just as with the IACUC committee, she said. Fair suggested that biosafety professionals should be trained in understanding the risk of research in the field with wild animals and the trade-offs of risk in the field. Personal protective equipment should be appropriate for each situation, but not hinder wildlife professionals or add additional risk for both researchers and animals.

The techniques for capture and handling will vary greatly depending on the species involved. Fair said that it is important for researchers to know the best practices for their species and how their species may be sensitive to being captured. It is useful to think through secondary or indirect impacts on birds such as increasing predation on a bird or nest. The types of euthanasia used for birds in the field will also vary for the species and field situation, she added. Fair noted that researchers and IACUCs must work together to determine the best euthanasia techniques and researchers must be ready to use these techniques in an emergency with an injured bird.

To summarize, Fair offered a few suggestions for the improvement of wild bird research and the collective understanding of the challenges of work in the field with wildlife. The goal of researchers, IACUCs, and biosafety professionals is ensuring the ethical treatment of animals, using appropriate

biosafety protocols, and fostering high-quality science that increases understanding of the natural world. Fair’s suggestions include encouraging more wildlife experts and PIs to serve on IACUCs; providing training opportunities for researchers, IACUCs, and biosafety professionals; encouraging the use of wildlife specific guidelines such as the Guidelines to the Use of Wild Birds in Research (Fair et al. 2010); and attending more IACUC wildlife 101-type workshops. Lastly, Fair encouraged the publication of relevant data on the impacts of research on birds, especially if it is a negative impact. Researchers continue to innovate new and better ways for investigating wildlife, Fair said, and the future holds even more promise for continuing to learn about the variation and natural history of birds and the understanding of populations and the possible mitigations for conservation.

CAPTURE AND HANDLING OF FREE-LIVING WILD SMALL MAMMALS FOR RESEARCH AND EDUCATION

George James Kenagy has been with the University of Washington since 1976 where his research and teaching with undergraduate and Ph.D. students has involved work with many diverse kinds of small mammals. He has worked in North and South America, Australia, Asia, and Europe on research at the interface of ecology, with behavior, physiology, and evolution. Kenagy said that the key to his work has been to capture and handle animals in their natural habitats and to release them for subsequent recaptures in the study of their seasonal activity and reproductive and life histories. Doing that, said Kenagy, is vitally important, especially in teaching field courses and in order to maintain or discover more of the remarkable diversity of lifestyles where they have evolved in nature. Kenagy said that they bring those same species into the laboratory to study their behavior and physiology under controlled conditions and also collect specimens for the Burke Museum at the University of Washington, where they serve as morphological vouchers and provide tissues for genetic analyses.

Kenagy said that small mammals are the most easily captured and handled in the field. It is easy, he said, to set out hundreds of small, live traps with a handful of bait that provides more than a day’s energy supply as a reward for the smallest of rodents. Individually marked animals return repeatedly over months and years to be re-trapped. In cold seasons of overnight trapping for nocturnal rodents, cotton balls are included along with the bait and the animals create a nest for insulation during their overnight lodging, Kenagy said. The animals can be removed directly from the traps by hand in many cases with species that are not inclined to bite. Day active small mammals, such as squirrels, are often trapped for just a few hours in the morning to avoid exposure to the Sun’s heat through traps that are typically shaded for protection. In some habitats, one can catch as many as six different species, Kenagy explained.

Many species, especially when they weigh less than 50 grams, are easily held between the thumb and forefinger by the scruff of the neck for examination and identification of sex and reproductive condition, which is done in a field course to introduce students to small mammal populations. Kenagy added a series of examples to illustrate his point. He said that a kangaroo rat can be identified by its unusual lower incisors and the expansive external fur-lined cheek pouches characteristic of this rodent Family. For ground squirrels, researchers can identify males, as their testicles can be palpated and measured. Animals such as deer mice remain calm when held gently and once released they quickly reorient and scurry away into safety. It is easy to weigh small mammals with one person and without the need of an immobilizing inhalant, Kenagy said. The animal enters a bag from the trap and the bag is suspended from a spring balance.

For physiologically oriented field studies, Kenagy said, animals will often receive injections; for example, water with stable isotopes oxygen-18 and deuterium to measure energy and water metabolism; or injections of oxytocin to induce milk letdown for the micro-sampling of milk. Blood samples are taken for metabolic and hormonal studies. Another minor procedure is attaching radio telemetry collars. Some of these procedures may be more easily carried out using a brief exposure, on the order of 10 to 20 seconds, to an inhalant anesthetic in a jar carried along in the field as each trap is checked. A freshly captured kangaroo rat can be held briefly in an aquarium and photographed from below using an angled mirror enabling observation of how it processes the special leaves it eats. Plastic jars are adequate as

holding containers or for brief transport but can be used for more extended transport to the laboratory or for shipping by adding bedding and food. Kenagy said that he has taken two species of desert rodents in these jars on an airline flight to Germany to a laboratory where he studied their circadian rhythms for a couple of years.

Kenagy outlined data he acquired by studying small mammals in the field. For example, in a 3-year study in the Northern Mojave Desert of three Heteromyidae rodents—two bigger kangaroo rats and a tiny pocket mouse that weighed only 6 grams (Kenagy 1973)—Kenagy was able to obtain data by repeated monthly sampling to document annual cycles of recruitment and population abundance. Recorded annual cycles of body mass and differences in the breeding periodicities of the two kangaroo rat species reflected differences in their diets and water balance physiology.

Kenagy also conducted an expanded study of seasonality reproduction in response to desert environmental conditions that involved five rodent species from the same area over three breeding seasons with several consecutive days of trapping every month (Kenagy and Bartholomew 1985). They also began to study reproductive function in the laboratory by bringing freshly captured individuals in to set them up under various experimental variables, such as diet, dietary water content, and day length. These are animals that do not regularly drink water where they live in the desert (Kenagy and Bartholomew 1985). The five species responded differently based on body size and diet and variously produced litters that varied from only one to as many as 11 pups; and mating seasons that varied from 1–6 months depending on the species and each year’s particular environmental conditions. In some species’ cases, they found years of complete reproductive failure.

Kenagy’s most extensive research studies, lasting nearly a decade, were on a single population of golden-mantled ground squirrels in the Washington Cascades, which breed just once per year and hibernate for about 6 months. Their theme was seasonal variation in energy expenditure and allocation to reproduction (Kenagy et al. 1989, 1990; Boswell et al. 1994). With 275 traps on a 22-hectare grid, they trapped for many consecutive days during mating, gestation, and lactation with a team of three to five members. By weighing females frequently, they determined dates of parturition and mating by backdating through gestation. They counted the number of pups by x-raying females in late gestation, and then observed the number that emerged upon weaning. This kind of field study is labor and time intensive, and the squirrels were amenable, clearly habituated to being trapped and observed, something not necessarily possible with all small mammals, Kenagy said. The simplified summary of a general monthly energy budget shows low monthly expenditures during hibernation, followed by the launch of the breeding season, with males spending much more energy moving around than the females who emerge later in estrus. From then on, the female budget exceeds that of the males because of gestation and, even more demanding, lactation to support the growth of the young.

Kenagy emphasized the importance of regular trapping to monitor the animals and perform special procedures. The patterns of body weight demonstrated the remarkable differences between the sexes in energy allocation. Females emerge from hibernation at the lowest weight of the year, accelerate their feeding behavior to support production of the young, and then fatten to double their beginning weight for hibernation. Males maintain a high body weight all year and defend an accumulated food cache in their burrows that allowed them to emerge from hibernation in top condition for mating competition. Data from the stable isotopes that determine metabolic rate, measured by an initial injection and blood sample, followed at 2–3 days by a second blood sample, gave a reading of total water metabolism and daily energy expenditure. During mating the male spends much more energy than the female, about 2.5 times basal metabolism. During lactation, however, an average female energy budget is much higher, more than three times basal.

Kenagy’s research team also studied glucocorticoids in free-living, female, yellow-pine chipmunk, a smaller and even easier to handle hibernating squirrel (Kenagy and Place 2000). They measured hormones across the active season, as in the ground squirrels, and they also made a special assessment of the effects of capture and handling (Kenagy and Place 2000; Place and Kenagy 2000). The biggest seasonal increase found was a four-fold increase in corticosterone in females during lactation, which is basically the same result as in the ground squirrels. The chipmunks were captured repeatedly

during the season in the small box traps where they remain quietly in the morning for a few hours before blood samples were taken. Apparent effects of capture and handling on the glucocorticoids of chipmunks were measured by setting a few traps closely together in a small space that they could be visually monitored from a single vantage point to determine the precise moment of capture. By running to the trap and quickly carrying out the usual blood sampling including 10–20 seconds of exposure to inhaling anesthesia, blood was obtained within 2–3 minutes of capture. Each animal was returned to its trap, and after waiting 30 minutes a second blood sample was taken. Normal chipmunk behavior of stuffing cheek pouches with bait seeds while in the traps for 30 minutes was observed. Glucocorticoids consistently increased above the initial values, ranging from 50% increase to more than double, depending on the season and the hormone. Increases were interpreted as a response to capture and handling, including the initial blood sampling, which seems to be a valid demonstration of a glucocorticoid response to a special energy demand. From the short duration of 30 minutes of this acute response, chronic negative conditions or harm resulting to the animals were not observed.

Kenagy mentioned earlier the value of depositing novel and diverse specimens in trusted research museums and identified some significant genetic geographic variation within and between species. New patterns of historical change in genetic population structure, the speciation process, and even hybridization have been uncovered through their research studies (Whorley et al. 2004; Himes et al. 2008; Chavez et al. 2014). Such studies are important not only as contributions to evolutionary biology but also for conservation efforts that may apply to local populations or entire species. Kenagy highlighted some of the advantages of conducting research in the field on small mammal populations that are abundant and accessible and studied with little need for special restraining or handling techniques simply due to their small size. In closing, Kenagy advocated for the important educational role of promoting in-the-field live trapping demonstrations of small mammal populations for college and university students.