Summary

Well-written procedures provide a detailed and complete description of the animal’s experience throughout the procedure, isolating procedural details from experimental context.  For more complicated procedures, a chronological list works best.

Tips for what to include:

• For more complex procedure types such as surgeries, imaging and irradiation, behavioral tests:
  • include any substances or types/classes of substances. For example, if you use a contrast agent for imaging; if you use a standard antidepressant or anxiolytic as a control for a behavioral test; if it is a surgery procedure, add in your anesthetics and analgesics, including the name and dose and time/frequency of administration as part of the procedure (see Procedure 1 Surgery example below); 
  • frequency and approximate duration of the length of procedure (always best to include a range of time or a maximum duration, using language such as “up to X hours”. An example is "up to 5 times over a period 20 days with a minimum of 48 hours between injections.")

Surgery specifics:

• survival or non-survival 
• approximate length of incision, how it’s closed
• implant information (e.g., type, size), if applicable
• implant content (e.g., estrogen pellet), if applicable and key to the model 
• monitoring (during and post-surgical)

Substance administration specifics:

• route of administration
• is the substance available in pharmaceutical grade? 
• any anesthesia when administering substance?
• or
• restraint type and duration
• is monitoring needed post-administration?

Behavioral testing specifics: 

• session duration 
• stimuli (note: justification for any aversive stimuli, if project-specific, should be excluded as that would to be evaluated by each institution’s IACUC)
• food or fluid regulation
• restraint type and duration
• number of times any one animal will be subjected to the test
• interval between sessions of this test on any one animal
• any training or habituation required
• positive reinforcement
• repetition 
• order (if multiple test types within this procedure). 
• testing chamber (size, material) and cleaning frequency (e.g., between each animal, end of the day) and what cleaning solution is used

Capture and Trapping specifics:

• type of environment (e.g., marsh, river, ocean, forest, plains, etc) 
• equipment being used (e.g., mist nets, trawls, seining, boats) and cleaned or sanitized
• any limitations due to being in the field (e.g., veterinary care if there is an injured animal) 
• if animals are captured/trapped, include details about how animals are restrained and how long animals are studied before release, retention, or euthanasia. 

Procedure Examples

Procedure Example 1: Surgery

PROCEDURE DESCRIPTION

Duration: up to 120 minutes

Surgical Preparation of Animal: No restrictions on food or fluid prior to anesthesia and surgery. Once animals are anesthetized, ophthalmic ointment will be applied. Then, the skin at the surgical sites at the skull will be shaved and prepped with 3 alternating scrubs of alcohol and betadine.

Surgical Preparation of Surgeon: All standard aseptic precautions (use of headcover, facemask, sterile gloves, gown).

Instruments, Method of Sterilization (prior to first surgery and between animals if multiple surgeries conducted sequentially): Autoclave or chemical sterilization of instruments prior to first surgery, hot bead sterilize instruments between animals for a maximum of 5 surgeries.

Analgesia: Meloxicam (1mg/kg, SC, 24 hr) will be given at time of procedure. Carprofen (5mg/kg, SC, 12 hr) will be used as a substitute if meloxicam is not available.

Surgery Description:
Animals anesthetized with 1-5% isoflurane are placed in the stereotaxic device. Animals are monitored for absence of withdrawal response to mild paw pinch prior to incision. Once a surgical plane of anesthesia is confirmed, the animal is prepped as described above. A ~1-cm incision is made using standard sterile-tip procedures. The skull surface anatomy is used to identify the site of cranial drilling using a stereotaxic device. Up to 2 small bore hole(s) are drilled in the skull above the targeted site(s).

For injections, a needle (e.g., 27g), which is sterilized between surgeries using a hot bead sterilizer, is connected to a Hamilton syringe and controlled by a microprocessor driven pump and lowered to the position of injection. Injection (generally 1-3 μl of drug or viral particles) is given at a rate of 1μl per 5 min, the needle is then slowly withdrawn. The surgical wound is closed using sutures, skin adhesive and/or staples.

For cannulation or implantation of optic fibers or optrodes (i.e. optical sensor device), a sterilized stainless steel guide cannula or an optic fiber or optrode will be inserted into targeted brain regions and 2-4 skull screws will be affixed to the skull. The guide cannula or optic fiber or optrode will be fixed into place with dental cement and a dummy cannula will be inserted into the cannula to keep debris from entering. Some animals may receive viral infusions in an earlier surgery or sequentially in the same surgery prior to implantation of a cannula, optic fibers or optrodes.

Surgical Monitoring: Respiratory rate, toe pinch (anesthesia depth) with thermal support

Surgical Closure:
- Skin, Suture Type: Nylon or monofilament absorbable (adhesive or staples also an option). If implanting cannula, head caps are made with skull screws and dental cement to fix the cannula into place; this head cap covers the incision site so no surgical closure is needed.

Post-operative care procedure and monitoring:

Animals are provided with thermal support during recovery from anesthesia and transferred to their home cage for general housing once they appear alert, mobile, and are grooming. Animals are observed daily for at least three days post-operatively and then weekly thereafter to assess for signs of surgical complication, pain or poor health as described below.

Signs of surgical complication include wound dehiscence, incisional redness or discharge, severe lethargy and/or immobility and neurological signs such as circling, head pressing, head tilt, and abnormal respirations. Veterinary Services will be contacted immediately if any of these signs are observed.

Animals will also be monitored for more general indicators of pain or poor health such as hunched posture, poor grooming, porphyrin staining, mild lethargy, and mild weight loss and/or decreased body condition score. If these signs are observed 24-48 hours after surgery, Veterinary Services will be consulted.

Sutures or wound clips will be removed 7-14 days after surgery.

Special considerations:

For a repair surgery, if the skin at the surgical site reopens, following preparation of the animal as described above, sutures (or adhesive or staples) will be applied in order to close the surgical site. A maximum of one repair surgery will be performed per animal.

KEYWORDS: cranial, cannula, optic fibers

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Procedure Example 2: Husbandry and Veterinary Medicine

ID: 168 (automatically assigned by CUSP site) 

Procedure Name: 

Procedure Type:        

Species:            

Contributing Institution:    University XYZ

Date Submitted:         October 10, 2024

Date Last Modified:        November 6, 2024

Parent Procedure:        None

PROCEDURE DESCRIPTION

Cage size 

The minimum cage size for a hummingbird is 1.5 ft wide x 2.5 ft long x 1.5 ft high, and hummingbirds should ideally be singly housed.

Outdoor cages 

If housing hummingbirds outdoors, cages should have mesh bottoms to keep out predators and adequate shade in the summer. Outdoor cages are not recommended for the winter or harsh climates. 

Special considerations:

Hummingbirds are territorial and are unlikely to be attracted to hummingbird houses. Environmental enrichment should provide platforms they can use to nest on. Hummingbirds generally place their tiny cup-shaped nests on top of branches or even on human-made supports like windchimes, faucets, and string lights.

KEYWORDS: Trochilidae, enrichment, cage size

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Procedure Example 3: Blood or Sample Collection & Identification and Genotyping

ID:                229

Procedure Name:        Fin clip

Procedure Type:        Blood or Sample Collection & Identification and Genotyping

Species:            Zebrafish

Contributing Institution:    University XYZ

Date Submitted:         February 10, 2024

Date Last Modified:        March 31, 2024

Parent Procedure:        None

PROCEDURE DESCRIPTION

• Fin tissue is collected from fish at 2dpf-2yr for genotyping, histological analysis or biological analysis.. Fish will be anesthetized in buffered MS-222 (50-200 mg/L) and caudal fin tissue removed using a brand new single-use, single-sided razor blade under the microscope. The total amount of time in anesthetic is typically < 10 minutes. Fish are then immediately transferred to a container with fresh system water and monitored continuously until they recover the ability to right themselves. Upon immersion in fresh water fish typically regain swimming ability within 5 minutes. 

•Less than 50% of the tail fin will be removed. Although individuals will typically be subject to this procedure only once, in some cases to obtain sufficient DNA sample from genetically unique individuals for the purposes of gene mapping while allowing individuals to be kept for natural breeding, fin clip of an individual may be performed up to 5 times with sufficient time between fin clipping for complete regeneration of the fin to occur (typically at least one-two weeks between fin clipping). 

•Fish wounds spontaneously and rapidly heal. Following the procedure, animals will be monitored by research staff for up to 3 days following the procedure or until proper swimming-righting and feeding behaviors have resumed. Animals showing signs of necrosis, infection or distress will be euthanized.  

KEYWORDS: Danio rerio, DNA collection

PROCEDURE NAME Conventions/Recommendations

-Best NOT to include in the name: species or procedure type as these are redundant

-DO include any descriptor that further specifies how the procedure is done (e.g., route of administration or sampling, what type of euthanasia, the purpose of the surgery and the organ/tissue, type of implant, method of capture, etc).

-Start broad and get more specific; can use a comma or colon to add more information

-At the same time, keep it simple and use as few words as possible

Examples:

Euthanasia:    Isoflurane Overdose Followed by Cervical Dislocation, Open Drop Method

    Decapitation, Under CO2 Anesthesia (<14 days of age)

    Exsanguination via Cardiac Puncture, Under Tribromoethanol (Avertin) Anesthesia

Surgery:            Non-Survival, Organ Harvest

Non-Survival, Perfusion, Cardiac, No Fixation, Under Ketamine/Xylazine Anesthesia

                Survival, Muscle Implant Surgery

                Survival, Osmotic Minipump implantation

                Survival, Intracranial Injection Under Isoflurane

Substance Admin:         Staphylococcus aureus Infection    

                Neural Stem Cells Graft    

                Anesthesia, Isoflurane, Short Duration (<1 hour)

                Anesthesia, Terminal, Pentobarbital or Pentobarbital Solution

                Analgesia, Meloxicam (SC, 24 hours)

                Glucose Tolerance Test, IV

Imaging and Irradiation:    Echocardiography

                Infrared Thermographic Camera

                Irradiation, Sub-Lethal, for Adoptive Cell Transfer

                Irradiation, Lethal, for Bone Marrow Transplant

Blood & Tissue Collection:     Blood Collection, Tail Prick

                Tissue Collection, Skin Biopsy

Induction of Illness:        MPTP Model of Parkinson’s Disease

                Spinal Cord Injury, Percussion

                Tumor Transplantation, Human Prostate Cancer Cells

Diet Modification:        Semi-Purified Diets, Fat and Cholesterol Content Varied (Diets 1-5)

                Ensure Liquid Diet

Behavioral Testing:        Open Field Activity Monitoring

                Elevated Plus Maze

Husbandry and Veterinary

Medicine:            Octopus Dietary Recommendations

(Note: this category may include species)    Neonatal Ferret Care

                Armadillo Housing and Feeding

Other:                 Bladder Expression

Recording and/or Electrical Stimulation of Brain, Spinal Cord and/or Muscles