Pain/Distress Categories

• Raising for the purpose of producing or supplying laboratory animals
  • - Breeding and rearing of laboratory animals to secure experimental groups and maintain strains
  • - Pigment application
• Short-term restraint with a low degree of confinement for the purpose of observing appearance and behavior and conducting examinations
  • - Breeding and rearing to confirm phenotypes
  • - Restraint, measurement of body weight and temperature (without anesthesia), measurement of spontaneous activity, and behavioral observation (without anesthesia)
  • - Behavioral observation in the wild or in the absence of artificial stimuli
  • - Tail amputation of mice and rats aged 3 weeks or younger

The experimental procedures must be performed by a person with professional skills

• Observation of appearance and behavior, and clinical examination
  • - Individual identification procedures that cause little pain and do not induce deformity: ear punch, microchip insertion, tattoo, etc.
  • - Short-term restraint using a restraint device
  • - Short-term behavioral tests that can be performed without significant stress (maze tests, low-exertion stages of the Rota-Rod), non-invasive radiography, blood pressure measurement (tail cuffing)
  • - Short-term (within 8 hours) restriction of food and water
• Single-administration dosing and blood collection
  • - Oral, intragastric, intraperitoneal, intradermal, subcutaneous, and intramuscular administration of non-irritant substances that have no side effects
  • - Blood collection or catheter insertion using a route that causes minimal pain
  • - Major blood collection sites: cephalic vein, jugular vein, saphenous vein, tail vein
  • ※ Includes short-term restraint using a restraint device for blood collection or administration
• Cases in which euthanasia is performed after an infection experiment but before the onset of clinical signs
  • - Infection experiments in which no symptoms appear after infection
  • - Antibody production experiments that do not use adjuvants
  • - Experiments in which, even though a specific disease is induced, symptoms are very mild and the experiment is terminated early
• Education and practice
  • - Category B-level practice experiments performed by inexperienced personnel during the survival period of laboratory animals (limited to animal-use practice within a regular curriculum, where education and supervision are provided by a person skilled in animal experimentation techniques, and where the experimental method and content are already established and the procedure is simple)
• Non-survival experiments conducted after euthanasia
  • - Cases in which animals raised under Category B conditions must be euthanized because they are no longer needed for experiments or research
  • - Urine collection, skin biopsy (tissue sampling), and non-survival experiments conducted under deep anesthesia (post-mortem collection of body fluids or tissues)

• Cases in which physical, behavioral, and physiological changes are expected due to moderate or greater pain or confinement requiring the administration of drugs to relieve the animal's pain, and drugs to relieve pain (sedatives, analgesics, anesthetics) are administered
• Cases in which the substance administered to a laboratory animal is irritant, antigenic, corrosive, or harmful, so that pain or stress is aggravated after administration
• Blood-feeding experiments on laboratory animals by disease-vector insects
• Animal experiments that induce persistent itching or stress, such as atopic dermatitis
• Surgical procedures on laboratory animals (surgery and invasive procedures) generally performed under general or local anesthesia
• Long-term restraint using a restraint device
• Restriction of water and feed for 8 hours or more
• Forced exercise experiments
• Repeated intravenous blood collection
• Retro-orbital plexus blood collection or cardiac blood collection performed under anesthesia
• Biopsy of the tail, toes, etc.

• Administration of pain-inducing drugs
  • - Experiments administering pain-inducing substances (chemicals, toxic substances) that cause moderate inflammation or pain in the animal
  • - Antibody production experiments that use adjuvants
  • - Experiments that intentionally induce pain or inflammation, such as asthma, gastritis, colitis, and atopic dermatitis
• Infection, chronic disease, and pharmacological toxicity experiments
  • - Cases in which intentional infection or induction of a specific disease causes mild external symptoms but pain is expected due to metabolic and physiological changes
  • - Cases in which early-stage diseases such as tuberculosis, pneumonia, renal failure, colitis, diabetes, fatty liver, and cystitis are induced
  • - Cases in which no animals die after infection but moderate clinical signs such as fever and weight loss appear
  • - Disease-induction tests that cause severe pain and stress, such as irradiation to induce immunosuppression, and models of myocardial infarction, cerebral infarction, ischemia, spinal injury, nerve injury, Parkinson's disease, autoimmune disease, diabetes, hypertension, muscular atrophy, obesity and hyperlipidemia, and anticancer models
  • - Reproductive toxicity tests, single/repeated-dose toxicity tests, and pharmacological toxicity tests in which, based on a preliminary experiment, no animals are expected to die (however, if an animal dies, re-approval as Category E is required)
  • - Procedures that induce clinical signs such as anorexia, skin trauma, abscesses, lameness, conjunctivitis, corneal edema, and photophobia
  • - Induction of benign tumors or malignant tumors terminated before the metastatic stage. However, only when the tumor volume is 1 cm³ or less
  • ※ Even for a pre-metastatic-stage tumor, it must not significantly impair the animal's behavior
• Performing surgical procedures
  • - Refers to surgical procedures performed by a trained, skilled person that do not cause severe deformity or changes in bodily function after the procedure
  • - Includes general surgical operations and dental surgery
• Repeated blood collection and invasive examinations
  • - Restraint using a long-term restraint device for repeated administration and blood collection, and restriction of water and feed for 8 hours or more
  • - Tail tissue amputation, invasive blood pressure measurement, retro-orbital plexus blood collection, cardiac blood collection, and repeated intravenous blood collection
• Induction of physical stress
  • - Experiments that induce severe stress and maintain physical confinement for a long period
  • - Cases in which behavioral disorders are induced by the attachment of a semi-permanent cranial catheter
  • - Behavioral experiments that induce a moderate exercise load (e.g., treadmill)
  • - Behavioral experiments that include repeated electrical stimulation
• Diagnostic examinations under anesthesia
  • - Electroencephalography (EEG)
  • - Ultrasound examination
  • - Electrocardiography (ECG), etc.
• Raising for the purpose of producing or supplying nonhuman primates

• Cases in which extreme pain at or beyond the limit that a conscious animal can endure without anesthesia is inflicted
• Toxicity tests, tests inducing microbiological pathogenicity, irradiation tests, pain research, etc. performed under conditions in which pain-relieving drugs are not administered
• Orthopedic surgery, dental surgery, surgery inducing damage to cervical tissue or soft tissue, etc. performed under conditions in which pain-relieving drugs are not administered
• Cases in which persistent pain and confinement occur after a surgical procedure, even if drugs are used to relieve pain during the surgical procedure
• ※ When there is pain or distress but no anesthetic, analgesic, sedative, or tranquilizer is administered, the principal investigator must sufficiently justify to the committee the valid reason why pain-relieving measures cannot be taken (administration of an anesthetic for necropsy is unrelated to lowering the pain category)

• Administration of pain-inducing drugs
  • - Experiments administering pain-inducing drugs, chemicals, toxic substances, or organic substances that cause severe inflammation or pain
  • - Experiments that artificially induce disability by paralyzing animals or eliminating mobility through drug administration without anesthesia or long-term (several days to several weeks) restraint
  • - Irradiation intended to completely destroy bone marrow function
  • - Experiments that irritate the eyes or skin without pain relief, or experiments exposing animals to extreme environments
• Infection, chronic disease, and pharmacological toxicity experiments
  • - Cases in which the onset of symptoms and the death of animals occur in an infection experiment
  • - Cases in which intentional infection or induction of a specific disease leads to metabolic, physiological, and external changes that make normal life impossible
  • - Cases in which the death of laboratory animals is the experiment's endpoint
  • - Malignant tumors at the metastatic stage, terminal cancer (if the tumor volume exceeds 1 cm³ and causes significant impairment of behavior, it must be set as the experiment's endpoint and the animal euthanized)
  • - Cases in which the death of laboratory animals becomes the experiment's endpoint in pathogenicity tests or pharmacological toxicity tests
  • - Severe and terminal diseases such as tuberculosis, asthma, pneumonia, renal failure, colitis, diabetes, fatty liver, liver cirrhosis, and cystitis, or transplantation experiments accompanied by immune rejection reactions
• Performing surgical procedures
  • - Experiments that, even when performed by a trained, skilled person, cause severe pain due to surgical sequelae or permanent disability due to defects, etc.
  • - Surgery after which normal life is difficult
  • - Orthopedic surgery that induces bodily paralysis, loss of sensation, or functional impairment
  • - Surgery that induces cerebral ischemia, stroke, etc. causing semi-permanent damage
• Induction of physical stress
  • - Cases in which harmful stimuli such as shock are applied, or electrical stimulation at a level that a human would feel pain (1 mA or higher) is applied
  • - Behavioral tests that may cause moderate or greater fear and stress: long-duration swimming, water maze tests, etc.
  • - Experiments intended to cause pain: paw nociception confirmation experiments
• Changes in body weight or body temperature
  • - Dietary restriction and diseases that induce a body weight loss of 20% or more
  • - Cases in which body temperature drops by 5℃ or more
• Experiments using nonhuman primate monkeys
  • - Experiments other than those for breeding or supply purposes, etc.
  • - Separating nursing animals from their mothers
• Behavioral experiments accompanied by pain, such as the water maze