1. ANALGESIA, ANESTHESIA AND POST-OPERATIVE CARE IN LABORATORY ANIMALS Thiago Vignoli, PhD Médico Veterinário CRMV SP 20615

2. Procedures involving pain or discomfort to the animals, included euthanasia Essential knowledge: physical restraint/handling, routes of administration, appropriate equipment, vital parameters Anti-inflammatories, antibiotics, sedatives, tranquillizers, analgesics and anesthetics Copy or know that you are using! Initial Considerations

3. Choice of Technique Species-responses to agents Temperament and physical condition of the animal Familiarity with the animal Degree of invasiveness and duration of the procedure Prior experience with certain anesthetics Availability of equipment and drugs, as well as staff training

4. Fasting* Age Nutrition Stress Gestation PAM Usual medication Species/Animal strain Body and environmental temperature Invasive procedures, duration, etc. Factors Influencing Anesthesia

5. Surgical Procedure Steps

6. Reduce stress of handling, aggressiveness and fear Facilitates the preparation for surgery and other procedures Reduces anesthetic dose and their undesirable effects PRE-ANESTHETIC MEDICATION Preparation laboratory: surgical field, anesthetics and solutions Animal Care: housing, handling, drug administration (Respect!) Environmental Care: Asepsis, noise, movement of people, light and odors

7. PRE-ANESTHETIC MEDICATION 1)ANTICHOLINERGICS - Parasympatholytic action; reduction of bronchial and salivary secretions (Flecknell et al., 1990; Harrison et al., 2006) Atropine 0,04 mg/Kg (SC, IP, IM) 2) TRANQUILLIZERS - Produce calming effect, drowsiness, reduced fear / anxiety, hypnotic, anticonvulsant (Pieri, 1981; Amrein, 1990) Neuroleptics: Acepromazine Benzodiazepines: Dizepan, Midazolan

8. 3) SEDATIVES -Produce sedation, hypnosis, muscle relaxation, ataxia, analgesia * (Murrell and Hellebrekers, 2005; Kastner, 2008) Alpha-2-adrenergic agonist: Xylazine, Medetomidine 4) OPIOIDS ANALGESICS -Produce moderate sedation and profound analgesia, without loss of consciousness (Breivik, 1994; Grape and Tramer, 2007) Fentanyl* (30 min), Tramadol (4-5h), Meperidine (1-2h), Sufentanyl (100x)  NSAIDs: Preemptive analgesia PRE-ANESTHETIC MEDICATION

9. ANESTHETIC INDUCTION AND MAINTENANCE 1)INJECTABLE ANESTHETICS - Produce unconsciousness (narcosis), neurovegetative protection, muscle relaxation, mild analgesia (Glen and Hunter, 1984; Murayama et al., 2005) Ketamine / Tiletamine (dissociative) Barbiturates (peritonitis), Propofol (without cumulative effect / exclusive IV) 2)INHALATIONAL ANESTHETICS - Globally depress the CNS (no analgesia) - Great advantages! - Orotracheal intubation (cannulae or scalpe PE) or mask - Vaporizer: Induction 5% and Maintenance 1-3% Halothane*, Isoflurane, Sevoflurane

10. Ketamine (60 a 80 mg/Kg) + Xylazine (10 a 30 mg/Kg) ANESTHETICS AGENTS IN ASSOCIATION Diazepan (2 mg/Kg) ou Acepromazine (1 mg/Kg) ou Tramadol (2 mg/Kg) Ketamine (40 a 60 mg/Kg) + Xylazine (10 a 15 mg/Kg) Flecknell, 2009; Unifesp, 2005

11. Physiological Monitoring Responses to Painful Stimuli: pedal withdrawal reflex! Ophthalmic ointment or liquid tears to prevent damage to the cornea

12. VAP VDI BZP, NLT OPIOIDS GENERAL ANESTHETICS XYLAZINE KETAMINE

14. POST-SURGICAL: PAIN 1)NON-STEROIDAL ANTI-INFLAMMATORY: Dipirone (10-20mg/Kg, VO, SC / BID or TID) and Acetominophen (200-300 mg/Kg, VO / BID or TID) Ketoprofen (2-5 mg/Kg, VO, SC / SID) Carprofen (5 mg/Kg, VO / BID) Hawk and Leary, 2005 ; Unifesp, 2005

15. POST-SURGICAL: PAIN 1)NON-STEROIDAL ANTI-INFLAMMATORY: Meloxicam (1-5 mg/Kg, VO, SC, IM / BID) Flunixin Meglumin (1 - 2,5 mg/Kg, VO, SC / SID) 2) OPIOIDS ANALGESICS : Tramadol (5 mg/Kg, VO, SC, IM / SID or BID) Hawk and Leary, 2005 ; Unifesp, 2005

16. POST-SURGICAL: INFECTION 3) ANTIBIOTICS: Enrofloxacin (5-10 mg/Kg, VO, SC / BID) Ampicillin (100-150 mg/Kg, VO, SC / BID) Sulfamethoxazole/Trimethoprim (15-30 mg/Kg, VO, SC / BID) Hawk and Leary, 2005 ; Unifesp, 2005

17. POST-SURGICAL: INFECTION 3) ANTIBIOTICS:  Pregnancy and Lactation Penicillin (100.000 UI/Kg, SC, IM / SID) Cephalexin (60 mg/Kg, VO / BID) Hawk and Leary, 2005 ; Unifesp, 2005

18. House them separately after surgery (blood / cages) Low light, noise and manipulations Temperature: 27-30°C adults and 35-37°C neonates until restore parameters (mice!)

19. Administration of SC or IP fluids Evaluate the parameters of pain and stress!!!

20. EVALUATIN OF GENERAL ACTIVITY Unrest Movement reduction Abdominal writhe PHYSIOLOGICAL CHANGES RR/ HR Body temperature APPEARANCE Piloerection Porphyrin Back arch Weight loss BEHAVIOR Aggression Social interaction reduced VOCALIZATION CHANGE IN EATING BEHAVIOR Food and water comsuption Urine/Faeces EVALUATION OF SURGICAL SITE Edema Licked Points PARAMETERS OF EVALUATION PAIN/STRESS PARAMETERS OF EVALUATION PAIN/STRESS Stasiak, et al., 2003

21. Facial Expression of Pain (Grimace Scale) Dr Mary Lynch (president of the Canadian Pain Society), Nature 2010; Sotocinal, et al., 2010, 2011

22. (Keating, 2012; Flecknell, et al, 2012) Orbital tightening Cheek flattening Nose shape Whisker position Ear position

23. Piloerection

24. Porphyrin

26. “ All species, including humans, feel pain with the same quality and intensity ”

27. TODAY IT’S A MORAL AND ETHICAL DUTY OF MAN TO MINIMIZE ANIMAL SUFFERING (WHO, 2006)

28. Thiago Vignoli, MV, PhD thiagovignoli@alesco.ind.br Tel: (11) 99715-5508

47. If you were to ask any of the monkeys why they were knocking other monkeys that tryed to pick up bananas, maybe the answer would be: “I do not know, things have always been so here!” The conformism and inertia make human being limited by choice.

48. “The mind that opens to a new idea never returns to its original size.” ALBERT EINSTEIN

49. Manual de Terapêutica Veterinária (3° ed), Silvia Franco Andrade, 2008 Anestesiologia Veterinária – Farmacologia e Técnicas (6° ed), Flávio Massone, 2011 Princípios Éticos e Práticos do uso de Animais de Experimentação, Universidade Federal de São Paulo, 2005 Laboratory Animal Anaesthesia (3rd edition), Paul Flecknell, 2009 Formulary for Laboratory Animals (3rd edition), C. Terrance Hawk and Steven L. Leary, 2005 Canadian Council on Animal Care (CCAC) REFERENCES