Appendix A Biomedical Protocols.docx

APPENDIX B: BIOMEDICAL PROTOCOLS

BIOMEDICAL PROTOCOL FOR FREE-RANGING CERVIDAE IN MONTANA:

Capture, anesthesia, tagging, sampling and necropsy procedures.

Wildlife Health Program 1400 S. 19th Bozeman, Montana

General Capture and chemical immobilization of free-ranging Cervidae should be carried out by a team of professionals with proper training, experience, and expertise in wildlife capture, veterinary anesthesia and animal handling. Capture data should be collected on the standard Wildlife Immobilization Form.

Legal Considerations The purpose of the Montana Fish Wildlife and Parks Animal care and Use Committee (FWP-ACUC) is to facilitate utilization of free-ranging wildlife in Montana for scientific study in accordance with the United States Department of Agriculture Animal Welfare Act. To this end, all requests (internal and external) for scholarly study of wildlife in Montana must be submitted to and approved by the FWP-ACUC.

Physical Immobilization Several forms of physical immobilization have been effectively used to safely and effectively capture cervids. Net guns, drive nets and clover traps have been used to immobilize large numbers of deer and elk although other methods do exist. Net guns have also been used effectively on moose. When selecting a method of physical immobilization human safety, animal safety, species to be captured and type of terrain must be considered.

Chemical Immobilization Adult deer, elk and moose may be immobilized by darting them from the ground or from a helicopter. The most common complications encountered during anesthesia are hypoxemia, hyperthermia and bloat. Capture stress and/or capture myopathy are potentially serious complications that can very difficult to treat in a field situation. Treatment is often unsuccessful.

Carfentanil or thiafentanil, often in combination with xylazine are generally the preferred drugs for immobilization of cervidae. The drug dosage rates will be determined by the prescribing veterinarian.

Additional doses for immobilization Animals that do not become recumbent within an appropriate amount of time after darting should be re-dosed according to instructions by the prescribing veterinarian.

Adjunctive Therapy At the discretion of the veterinarian, animals that are injured as a result of the immobilization process may receive prophylactic antibiotic therapy. Withdrawal periods must be observed (as stated in the Prescription Drug Acquisition and Use Protocol) in animals that may potentially be hunted for food.

Handling of immobilized animals Once the animal is recumbent it should be approached with caution and as with as little noise as possible. Eye covers should be placed on the animal immediately; they act as an additional means of restraint, protect the eyes and can prolong and improve the effects of immobilization. Monitoring of vital signs should begin as soon as possible after recumbency. Respiration and oxygenation are the most critical indications of an animal’s well being under anesthesia and pulse oximetry should be used as an adjunct to monitoring whenever possible. Cardiac monitoring is also important especially in animals immobilized by one of the agents that can cause bradycardia or hypotension. Temperature monitoring and control is also important. Animals generate a significant amount of heat during the exertion of capture and once immobilized have no means with which to dissipate it.

Baseline body temperatures (BT), heart rates (HR) and respiratory rates (RR) have been recorded from captured moose, deer and elk. Excitability, season and drugs are known to influence the values, excitability being the most influential factor. Safe expected ranges are: BT 101 – 103oF (38.4oC to 39.5oC), HR 60 to 90 beats/min (b/min), and RR 10 to 40 respirations/min (r/min). These ranges incorporate all seasons and the use of a central nervous system depressant drug. Values considered critical and an indication that corrective action should be taken include: BT >104oF (40.2oC), HR 102 b/min, and RR 40 r/min or <4 r/min. Persons trained in monitoring vital signs should be present during capture and immobilization procedures.

In warm weather, elect to immobilize animals in the cooler periods during the day and use water to wet the animal down to increase cooling. Cervidae are prone to hyperthermia. In very cold conditions, be vigilant for evidence of hypothermia and be prepared to respond accordingly. Immobilized ruminants are best placed in sternal recumbency to reduce the complications of bloat and regurgitation. However, some animals must be placed in lateral recumbency. The head should be elevated slightly above the level of the rumen to prevent regurgitation and the mouth should be slightly lower than the neck to allow saliva to drain. Ensure that the nostrils are clear and that the animal is breathing without difficulty. Remove the dart and needle and clean and treat the dart wound as instructed by the veterinarian prior to reversing the immobilization and releasing the animal.

Euthanasia Consistent with AVMA Panel on Euthanasia (2000) and as specified in the FWP Euthanasia guidelines. The veterinarian responsible for oversight will establish a protocol for euthanasia.

Tagging and Sampling All animals will be ear-tagged using an identifying MFWP plastic ear tag with a unique identifying number and the following printed on the back: “Call Before Eating”.

Moose, elk and deer may be captured for radio-tagging or sampling purposes and should be processed according to the aim of the project. In the case of moose, rapid response for urban problem animal control may require the capture and translocation of individual animals with minimal preparation time. Radio collars (VHF, GPS or Satellite) should be fitted according to the size, age and sex of the animal.

If needed, body measurements should be recorded according to established protocols. Blood is collected from the jugular vein by needle and syringe or using the BD Vacutainer® system. To facilitate sampling, blood should be collected immediately after capture of the animal.

Other biological materials should be sampled according to study protocol.

Necropsy Procedures In case of a capture-related mortality, the carcass should be transported to a veterinary diagnostic laboratory for a complete necropsy. (Wildlife Laboratory, Montana Department of Fish, Wildlife & Parks, Bozeman, Montana. Phone: 406-994-6357). As an alternative, an affiliated veterinarian or biologist can perform a field necropsy after consultation with the laboratory. To ensure rapid cooling of carcasses to be delivered to a laboratory, skinning the carcass can be considered. If transportation to the laboratory is not possible within 24-48 hours, the carcass should be frozen.

In the case of moose, always attempt to preserve the head and neck for laboratory examination for Elaeophora schneideri.

Biomedical protocol for free-ranging gray wolves (Canis lupus) in Montana: Capture, anesthesia, surgery, tagging, sampling and necropsy procedures.

Montana, Department of Fish, Wildlife and Parks

General Capture and physical or chemical immobilization of free-ranging gray wolves in Montana will be carried out by a team of professionals with adequate training, experience and expertise in wildlife capture, veterinary anesthesia, and animal handling. Capture data will be collected in accordance with the “Wolf Capture Sheet”.

Chemical immobilization Immobilization of wolves from helicopter and immobilization of trapped wolves may be performed using one of the following drug combinations. Drugs are administered via hand or remote injection:

1. Telazol® (Tiletamine + Zolazepam or TZ)

All animals > 6 months of age, regardless of sex and body mass, are darted with 500 mg (1 vial, reconstituted with 2.5 ml sterile water) Telazol® per animal. The drugs are administered by hand syringe, pole syringe or by a 3 ml dart syringe. The lightweight, plastic Tel-Inject® or Dan-Inject® dart is preferred for all thin-skinned animals but the PneuDart® and appropriately sized Palmer Cap-Chur® darts may be suitable where no alternatives exist. A 1.5 x 25 mm barbed needle is suitable for use in this species. To prevent the drug mixture from freezing, the needle can be filled with 70% ethanol before assembling of the dart. Atropine at 0.05mg/kg can be given to decrease salivation. This drug combination is safe and effective for immobilization of wolves however it is characterized by prolonged recovery times (~6-8 hrs for return of coordination and normal function) necessitating prolonged observation. In animals that are incompletely immobilized by TZ, additional administration of 1mg medetomidine (im or iv) is highly effective for deepening the plane of anesthesia. For reversal of immobilization in wolves that have been given medetomidine, animals should receive 5 mg of atipamezole (Antisedan®) i.m. or s.c. per mg of the total dose of medetomidine administered. Due to the long elimination time of TZ, atipamezole should not be given until 40-50 min after darting. In an emergency, atipamezole can be given at any time but recovery may then be rough with possible in-coordination, excitation and convulsions. Such an animal can be calmed by 5 mg midazolam (1 ml Midazolam®) i.v. or i.m.

2. Medetomidine, Ketamine, Atipamezole (or MKA)

All animals receive 5mg medetomidine + 250mg ketamine. The drugs are administered by hand syringe, pole syringe or by a 3 ml dart syringe. Reversal of effects of drug is effected by administration of atipamezole at 5mg per 1mg Medetomidine administered (i.e. 25mg for most animals). Antagonism of medetomidine allows recovery times (to coordinated ambulation) of ~40 mins. Disadvantages of this combination include cost and the potential for capture associated hyperthermia and shock.

3. Ketamine, Xylazine (or KX).

Remains the recommended drug combination for immobilizing wolves in North America. Administered at 10mg/kg ketamine + 2mg/kg xylazine (~350mg ketamine + 70mg xylazine per adult). Where required, supplementation with 5mg/kg ketamine (~150-200 mg per adult). Antagonism achieved using yohimbine at 0.15mg/kg (~5mg per adult, performed ~40 minutes after induction to allow for metabolism of ketamine).

*All animals should be observed by trained personnel until full recovery is evident. (This may take 4-6 hours in animals immobilized with 500 mg of TZ. Possible side effects and dangers during and immediately after recovery include vomiting, hyperthermia, hypothermia, attack from other wolves, open water, lack of fear, and traffic.)

Other treatment

· Captured animals with obvious health-threatening diseases should be treated according to accepted standards in veterinary medicine by or under the direct supervision of a veterinarian.

· Combined penicillin (benzathine and procaine penicillin) is administered to all animals at 15,000 – 20,000 IU/kg (~2 ml IM) following immobilization and handling.

· Dart wounds are treated with a broad-spectrum antibiotic or intra-mammary infusion (e.g. Cloxacillin dry cow therapy)

· Petroleum/mineral oil-based eye ointment is placed in both eyes at the time of fitting the blindfold.

· In animals with severe capture related injuries, terminal illness or extensive and generalized sarcoptic mange, euthanasia should be performed.

· Recommended treatment for generalized sarcoptic mange is 0.4 mg/kg of ivermectin s.c. (~ 1.0 - 1.5 ml of Ivomec® for an adult wolf).

· Vaccination of free-ranging wolves in Montana is currently not recommended.

Helicopter chasing and stress Wolves that have not been captured from (or chased by) a helicopter are usually naïve when approached and darting can be performed within a few minutes of observation if the snow condition and the area are optimal (ice-covered lakes, clear-cuts, open terrain etc). Wolves that have been captured before will usually run for cover when they hear the helicopter and are much more difficult to approach. To avoid stress and physiologic side effects (hyperthermia) during immobilization in such animals, intensive chasing should be kept to a minimum and the total time of discovery and pursuit should never exceed 30 minutes.

Handling of immobilized animals Immobilized animals should be monitored and clinically examined by the wolf specialist, biologist or wildlife veterinarian. Possible side effects include respiratory depression (drug overdose in individuals with poor body condition, aspiration of vomitus/saliva, pneumothorax due to misplaced dart), vomiting, and thermoregulatory dysfunction. If several animals are being captured at the same time, they should be brought together for monitoring and processing. To prevent aspiration of saliva or vomitus, immobilized animals should be kept in sternal or lateral recumbency with the mouth and head low relative to the body. An eye ointment should be applied to the cornea to prevent drying and eyes should be protected from exposure to direct sunlight (a blind-fold should be used whenever possible).

Thermoregulation should be monitored by frequent measurements of the rectal temperature (RT). “Normal” RT in wolves is thought to be 38.0-39.0°C (100 - 102oF).

· Hyperthermic animals (RT > 40.0 °C, 104oF) should be cooled by applying snow (or water in summertime) to the axilla, groin, and/or tongue. In case of persistent hyperthermia or RT > 41.0 °C (105.8oF), i.v. fluid therapy should be initiated (10-15 ml/kg/hr of Ringers Lactate or ~500 ml/hr in an adult wolf).

· Hypothermic (RT < 36.0 °C, 97oF) animals should be protected from wind and cold surfaces to avoid further cooling. In case of prolonged immobilization and recovery, hypothermic animals should be brought into a heated area and pre-warmed Ringers lactate (38 °C, 100oF) should be administered.

Cardio-respiratory function should be monitored using a pulse oximeter (Nellcor®) with the sensor (VetSat®, Nellcor) applied to the tongue. TZ (with or without the addition of medetomidine) has no major respiratory or cardiovascular side effects in gray wolves. A relative arterial oxygen saturation (SpO2) > 80 % is considered to be clinically acceptable in a field situation. A decreasing trend or SpO2 < 70% indicates inadequate ventilation and treatment with 5-10 mg/kg of doxapram i.v. (10-20 ml of Dopram®) should be considered. Where possible, a laryngoscope, endotracheal tubes and a ventilation bag should be available. Access to supplemental oxygen is recommended.

Tagging and sampling Most wolves are captured for tagging or sampling purposes and should be processed according to the aim of the project.

· Radio collars (VHF, GPS or satellite) should be fitted according to the size, age and sex of the animal. The transmitter (VHF) should be activated by removing the magnet and should be tested with the receiver before the animal is released.

· A microchip (AVID® or Trovan®) may be implanted s.c. at the base of the left ear. The microchip should be read with a portable scanner following implantation.

· Body measurements should be recorded according to the “Wolf Capture Sheet”. Photos, both whole body pictures and close-ups of fur, teeth, and eyes, should be taken of all animals (with identifying labels) and submitted to the designated wolf management specialist.

· Blood may be sampled from the cephalic (front leg) or the femoral vein (inside rear leg). 1 x 5 ml tube with EDTA as anticoagulant (LT or lavender-top Vacutainer®), and 2 x 10 ml tubes with clot activating factor (RT red top, or SST® serum-separator Vacutainer®) should be collected. 10% of the circulating blood volume (~ 60 ml/kg body weight) can be taken on a single occasion from a normal healthy adult with minimal adverse effect. A 30 ml syringe and 20 gauge, 1.5” needle may be used to collect sample. Blood for genetic studies should be collected (1 x 5 ml LT or whole blood from syringe, ‘spotted’ onto appropriate gene cards). Whole blood should be stored at minus 70°F; gene cards must be stored at room temperature and protected from extremes in light or humidity. Tubes without anticoagulant should be kept at room temperature for 1-2 hours to ensure complete coagulation. Serum should then be separated by centrifugation (1,500 G for at least 15 minutes) and transferred to 2 ml cryogenic vials (Nalgene®). Serum for banking (serology and back-up) is stored at minus 70°F. Banked blood, tissue and serum samples will be stored at the Wildlife Laboratory in Bozeman.

· Whole blood (LT) will be submitted to the Montana State Diagnostic Laboratory for a complete blood count with differential (CBC + Diff) and hemoparasite evaluation.

· Serum will be submitted to the Montana State Diagnostic Laboratory for testing exposure to the following pathogens:

· Infectious Canine Hepatitis virus (ICH)

· Canine Distemper Virus (CDV)

· Canine Parvovirus (CPV)

· Brucella abortus, Brucella canis

· Serovars of Leptospira interrogans

Where funds and sera are available, tests for exposure to the following pathogens will also be performed for surveillance purposes.

· Francisella tularensis (Tularemia)

· Coxiella burnetti (Q Fever)

· Yersinia pestis (Plague)

· Hair (2 x 15 ml sterile plastic tubes), skin biopsies (2 pieces of skin may be taken from the inside of the ear using a sterile 6 mm biopsy punch or from affected areas in animals with sarcoptic mange, and transferred to 2 x 2ml cryogenic vials), feces (1 x 50 ml sterile plastic tube), and other biological materials should be sampled according to the specific study protocol. Skin biopsies are preserved using 96% ethanol while hair and feces are frozen. These specimens will be stored at the Wildlife Laboratory in Bozeman prior to testing.

· Ectoparasites:

· Fleas: may be collected and placed directly in 70% ethyl alcohol for preservation and identification. Flea combs may be used to remove fleas and larvae from individual wolves.

· Ticks: may be removed from the animal and placed directly in 70% ethyl alcohol for preservation and identification.

· Mange mites: Definitive diagnosis (demonstration of mites) of mange is difficult and requires microscopic evaluation of deep skin scrapings. Sarcoptic mange is common in the free-ranging wolf populations of Montana and a presumptive diagnosis can be made on physical appearance of affected animals alone. In certain cases, the veterinarian may perform a skin biopsy. Affected animals may be treated using ivermectin administered at 0.4 mg/kg s.c. (NB. ivermectin is highly toxic to insects and fish and should be used with care). It should be noted however, that repeated exposure to sarcoptic mites might result in persistence of infection in the animal. Severely affected animals should be euthanized according to current AVMA guidelines (gunshot, captive bolt or lethal injection) and the carcass submitted to the Wildlife Laboratory in Bozeman.

Necropsy procedures In case of a capture related mortality, the carcass should be sent to the Montana FWP research laboratory for complete necropsy. To ensure rapid cooling, the carcass may be skinned. If transportation to the laboratory is not possible within 24-48 hours, the carcass should be frozen. As an alternative, a volunteer or contract veterinarian may carry out a field necropsy after consultations with the laboratory.

In general, all dead wolves found in the state of Montana are to be submitted to the Wildlife Laboratory in Bozeman for necropsy examination. In recently dead animals (within 24 hrs) a blood sample should be collected directly from heart (using 20ml syringe and 18 gauge, 1.5” needle inserted between the 3rd and 6th ribs on the left side of the thorax). The sample can be placed in an RT or SST® tube, spun in a centrifuge and submitted to the Wildlife Laboratory in Bozeman for storage or testing of serum.

In the event of a field examination being performed on a dead animal, guidelines detailed in Wildlife Necropsy Protocol should be followed. In addition, all reproductive tracts should be examined in detail. In females, evidence of pregnancy, recent breeding etc. should be noted.

For carcasses submitted to the Wildlife Lab, the following specimens and information will be collected:

· Body weight

· Color

· Field ID number

· Photographs (to be performed by attending wolf management specialist)

· Whole body

· Incisors (front view)

· Canines, molars (side view)

· Hide (Determined on a case-by-case basis according to directive, need and quality. Skinning will be performed by a wolf management specialist and disposition of the hide duly recorded. Where the hide is not collected, a record will be made detailing its disposal/destruction.)

· Skull (All skulls will be kept for further evaluation and will be appropriately identified and stored in the wolf freezer. Skull preparation, cleaning and storage will be performed by designated wolf management specialist)

· Tooth measurement and removal (All canines will be measured and results recorded on necropsy form. 1st premolar will be removed from all carcasses. Removal, cleaning and storage will be performed by wolf management specialist.)

· General necropsy examination. (Abnormal or suspect tissues will be submitted to the Montana State Diagnostic Laboratory for further evaluation.)

· Muscle tissue (Collected for DNA according to project guidelines.)

· Body fat index

· Kidney fat index

· Reproductive Evaluation

· Vaginal swab for cytology (estrus evaluation)

· Ovarian evaluation and CL count

· Fetus count

BIOMEDICAL PROTOCOL FOR FREE-RANGING BIGHORN SHEEP (Ovis canadensis) IN MONTANA: Capture, anesthesia, tagging, sampling, transportation and necropsy procedure Montana Department of Fish, Wildlife and Parks Wildlife Research Laboratory 1400 S. 19th Bozeman, Montana.

General Capture and chemical immobilization of free-ranging bighorn sheep should be carried out by a team of professionals with proper training, experience, and expertise in wildlife capture, veterinary anesthesia and animal handling. Capture data should be recorded on the standard Wildlife Immobilization Form. In Montana, adult body weights vary from ~70 kg (150 lbs) in females to ~110 kg (240 lbs) in males. Three month old lambs weigh ~23 kg (50 lbs). The rut typically occurs mid-November to late December. Gestation lasts 174 days, thus lambing usually occurs in the spring (mid-May to late June). Capture during the last trimester of pregnancy (mid-March onwards) should be avoided wherever possible.

Legal Considerations The purpose of the Montana Fish Wildlife and Parks Animal care and Use Committee (FWP-ACUC) is to facilitate utilization of free-ranging wildlife in Montana for scientific study in accordance with the United States Department of Agriculture Animal Welfare Act. To this end, all requests (internal and external) for scholarly study of wildlife in Montana must be submitted to and approved by the FWP-ACUC.

Physical Immobilization The net-gun has been found to have considerable advantages over the use of ground nets and chemical immobilization methods for capturing bighorn sheep. In a study by Kock et al (1987), the use of the net-gun resulted in the lowest proportion of compromised sheep at 11%, had no capture myopathy (CM) mortality, and resulted in a 2% accidental mortality. The use of drop-nets resulted in 15% compromised sheep, a CM mortality rate of 2%, and an accidental mortality rate of 1%. A similar proportion of sheep were compromised with drive-nets (16%). This method also had the highest CM mortality rate at 3%, and an accidental mortality rate of less than 1%. Chemical immobilization resulted in the most compromised sheep at 19%, had a CM mortality rate of 2%, and caused the most accidental deaths at 6%. Drop-nets and drive-nets were comparable when combining total mortality with rates for compromised bighorn sheep, 18% and 19%, respectively. Chemical immobilization had the highest combined measure of risk at 27% and net-gun lowest at 12%.

The use of blindfolds and hobbles is necessary to reduce stress and possible injury. Bighorn sheep should be kept sternal whenever possible. Handling, lifting or moving animals should be done in a manner that reduces the potential for injury to joints and the neck. Lifting animals by the head, neck or individual legs is not acceptable and may result in injury.

Bighorn sheep captured during net-gun operations and requiring the use of a helicopter to transfer sheep should be kept sternal. The use of a “transport bag” slung under the helicopter or placement of sheep inside the helicopter are both suitable options. Slinging bighorn sheep by hobbled legs and upside down may be necessary in certain situations but should be minimized to reduce the possibility of aspiration of rumen content. If required, slinging bighorn sheep upside down under a helicopter should be limited to distances of less than ½ mile.

Chemical Immobilization Bighorn sheep may be immobilized by darting them from the ground or from a helicopter. Anesthesia is similar to that of other ungulate species however careless use of immobilization drugs in bighorn sheep can contribute to hyperthermia, cardiac dysfunction, respiratory depression, lowered blood pressure, localized blood pooling, acidosis, bloat and aspiration. The most common complications encountered during anesthesia are respiratory depression, hyperthermia and bloat. Capture stress and/or capture myopathy are potentially serious complications that can be very difficult to treat in a field situation. Treatment is often unsuccessful.

A combination of carfentanil and xylazine, or a combination of thiafentanil and xylazine have been shown to produce reliable immobilization of bighorn sheep. Reversal of anesthesia can be achieved with naltrexone and tolazoline. Drug dosage rates will be determined by the prescribing veterinarian.

Withdrawal periods must be observed in animals that may potentially be hunted for food and animals must be tagged for future identification. Consult the FWP Prescription Drug Acquisition and Use Protocol for withdrawal periods.

Additional doses for immobilization Animals that are not recumbent after darting should be re-dosed as instructed by the prescribing veterinarian.

Adjunctive Therapy At the discretion of the veterinarian, animals that are injured as a result of the immobilization process may receive prophylactic antibiotic therapy. Animals captured for transplantation within Montana will be administered vitamin E, selenium, an antibiotic, such as Florfenicol, and drugs to remove parasite loads, such as Ivermectin. These adjunctive therapies may be administered to sheep captured for other reasons. Dosages of such drugs will be administered based on body size and recommendations stipulated on the vial. Changes to dosages may be made at the discretion of the veterinarian. Withdrawal periods of 30 days or more depending on drugs administered must be observed in animals that may potentially be hunted for food.

Handling of immobilized animals Once the animal is recumbent it should be approached with caution and with as little noise as possible. Eye covers should be placed on the animal immediately; they act as an additional means of restraint, protect the eyes and can prolong and improve the effects of immobilization. Monitoring of vital signs should begin as soon as possible after recumbency. Respiration and oxygenation are the most critical indications of an animal’s well being under anesthesia and pulse oximetry should be used as an adjunct to monitoring whenever possible. Cardiac monitoring is also important especially in animals immobilized by one of the agents that can cause bradycardia or hypotension. Temperature monitoring and control is also important. Animals generate a significant amount of heat during the exertion of capture and once immobilized have no means with which to dissipate it.

Baseline body temperatures (BT), heart rates (HR) and respiratory rates (RR) have been recorded from chemically immobilized bighorn sheep (Franzmann, 1971; Kock, 1987). Stress, exertion, ambient temperature and capture technique are known to influence the values. Safe expected ranges at capture are: BT 39.1oC (102.4oF) to 40oC (104oF), HR 125 to 130 beats/min (b/min), and RR 40 to 64 respirations/min (r/min). These ranges incorporate all seasons and the use of a central nervous system depressant drug. Values considered critical and an indication that corrective action should be taken include: BT 40.6oC (105oF), HR 145 b/min, and RR 75 r/min. Persons trained in monitoring vital signs should be present during bighorn sheep capture and immobilization procedures.

In warm weather, elect to immobilize animals in the cooler periods during the day and use water to wet the animal down to increase cooling. Sheep are prone to hyperthermia. In very cold conditions, be vigilant for evidence of hypothermia and be prepared to respond accordingly. Immobilized bighorn sheep are best placed in sternal recumbency to reduce the complications of bloat and regurgitation. However, animals often are maintained in lateral recumbency. As with all ruminants, the head should be elevated slightly above the level of the rumen to prevent regurgitation and the mouth should be slightly lower than the neck to allow saliva to drain. Ensure that the nostrils are clear and that the animal is breathing without difficulty. Remove the dart and needle and clean and treat the dart wound as instructed by the veterinarian prior to reversing the immobilization and releasing the animal.

Tagging and Sampling All animals will be ear-tagged using an identifying MFWP plastic ear tag with a unique identifying number and the following printed on the back: “Call Before Eating”.

On occasion, bighorn sheep may be captured for radio-tagging or sampling purposes and should be processed according to the aim of the project. Neck bands or radio collars (VHF or GPS) should be fitted according to the size, age and sex of the animal.

Body measurements, if needed, should be recorded according to established protocols. Blood is collected from the jugular vein by needle and syringe or using the BD Vacutainer® system. To facilitate sampling, blood should be collected immediately after capture of the animal.

Where feasible, pharyngeal/tonsillar swabs (using Dacron-tipped polyester culture swabs and Port-A-Cul transport media, available from the Wildlife Lab in Bozeman) should be collected for Pasteurella and Mycoplasma cultures.

Additional biological materials should be sampled according to specific study protocols and follow accepted procedures. Hair and skin biopsies may be preserved in 96% ethanol. Feces are kept cool but not frozen for parasite analysis.

Euthanasia Consistent with AVMA Panel on Euthanasia (2000) and as specified in the euthanasia guidelines of the FWP Prescription Drug Acquisition and Use Protocol. The veterinarian in charge of oversight will be responsible for establishing appropriate protocols for euthanasia.

Necropsy Procedures In case of a capture-related mortality, the carcass should be transported to a veterinary diagnostic laboratory for a complete necropsy or, as an alternative; an affiliated veterinarian or biologist can perform a field necropsy after consultation with the laboratory.

To ensure rapid cooling, skinning the carcass and opening the abdominal cavity (while preserving the integrity of the organs) can be considered. If transportation to the laboratory is not possible within 24-48 hours, the carcass should be frozen.

Long Distance Transportation If transportation of live animals is necessary it must be conducted in a manner that produces the least amount of stress to the animals. Bighorn sheep should be held in suitable trailers ambulatory and able to see. Trailers specifically designed for transporting sheep are preferred. These trailers generally allow for the separation of sheep into small groups of four or less. Standard horse trailers can be used for moving sheep but the insides of the trailers should be rounded with no square corners that allow sheep to congregate in one area. Floors of trailers should be lined with straw or other suitable material. All trailers should have adequate ventilation to allow for air transfer through the trailer, yet openings should be in locations or of small size to minimize the potential for injury to legs, heads or other body parts that may become lodged in openings. Adult rams (> 3 years old) should be separated from ewes and lambs when transporting. The maximum number of sheep in a trailer should not exceed 10 per 40 square feet of floor space (Foster 2005). Sheep held in trailers should be observed frequently but discretely to assess health status. Bighorn sheep should be transported as quickly as possible to release sites minimizing stay in trailers and reducing exposure to human disturbance. Additional information regarding capture and transportation of wild sheep is available in the “Wild Sheep Capture Guidelines” sponsored by the Northern Wild Sheep and Goat Council and Desert Bighorn Council (Foster 2005).

Literature

1: Kock MD, Clark RK, Franti CE, Jessup DA, Wehausen JD. 1987. Effects of capture on biological parameters in free-ranging bighorn sheep (Ovis canadensis): evaluation of normal, stressed and mortality outcomes and documentation of post-capture survival. J Wildl Dis. 23(4):652-62.

2: Kock MD, Jessup DA, Clark RK, Franti CE. 1987. Effects of capture on biological parameters in free-ranging bighorn sheep (Ovis canadensis): evaluation of drop-net, drive-net, chemical immobilization and the net-gun. J Wildl Dis. 23(4):641-51.

3: Kock MD, Jessup DA, Clark RK, Franti CE, Weaver RA. 1987. Capture methods in five subspecies of free-ranging bighorn sheep: an evaluation of drop-net, drive-net, chemical immobilization and the net-gun. J Wildl Dis. 23(4):634-40.

4: Franzmann AW, Thorne ET. 1970. Physiologic values in wild bighorn sheep (Ovis canadensis canadensis) at capture, after handling, and after captivity. J Am Vet Med Assoc. 1;157(5):647-50.

5: Franzmann AW. 1971. Comparative physiologic values in captive and wild bighorn sheep. J Wildl Dis. 7(2):105-8.

6: Foster CL. 2005. Wild Sheep Capture Guidelines. Biennial Symposium of the Northern Wild Sheep and Goat Council. 14:211-282.

IACUC Approval This biomedical protocol has been approved by the FWP Institutional Animal Care and Use Committee (IACUC) with the following stipulations:

1. Approval was granted for a five-year period ending December of 2012.

2. IACUC approval applies to management activities only. All research activities will require additional IACUC review.

3. Capture or handling activities that do not follow methodologies stipulated in this protocol will be done without IACUC approval.

IACUC attending Members:

Wildlife Division Approval Methodologies presented in this document are to serve as the guidelines for bighorn sheep capture, handling and transportation for management situations undertaken by Montana Fish, Wildlife and Parks. Variation from methodologies provided in this protocol should only be conducted under the guidance of the FWP Wildlife Veterinarian.

Ken McDonald, Wildlife Division Administrator Date