LABORATORY ANIMAL MEDICINE AND SCIENCE - SERIES II

LABORATORY ANIMALS: Rodent Anesthesia & Analgesia

V-9054

Kathleen A. Murray, DVM, MS Director, Technical Operations Charles River Laboratories, Inc. Wilmington, Massachusetts Cinthia Pekow, DVM Chief, Veterinary Medical Unit, Research Veterans Affairs Puget Sound Health Care System Seattle, Washington Gary L. Borkowski, DVM, DACLAM Attending Veterinarian & Associate Director Pharmacia Corporation St. Louis, Missouri

The Laboratory Animal Medicine and Science - Series II has been developed by the Autotutorial Committee of the American College of Laboratory Animal Medicine (ACLAM): C.W. McPherson, DVM, Chair; J.E. Harkness, DVM; J.F. Harwell, Jr., DVM; J.M. Linn, DVM; A.F. Moreland, DVM, G.L. Van Hoosier, Jr., DVM; L. Dahm, M.S.

Laboratory Animal Medicine and Science Series II is produced by the Health Sciences Center for Educational Resources University of Washington

LABORATORY ANIMAL MEDICINE AND SCIENCE - SERIES II

University of Washington Health Sciences Center for Educational Resources Box 357161, Seattle, WA 98195 -7161 206/685-1157 Copyright 2000 by the University of Washington Health Sciences Center for Educational Resources and the American College of Laboratory Animal Medicine All rights reserved. Printed in the United States of America.

V-9054 Lab Animal Medicine: Rodent Anesthesia & Analgesia

PRIMARY AUDIENCE SECONDARY AUDIENCE

Veterinarians, investigators, and research technicians. Veterinary students, laboratory animal care technicians.

GOAL

To provide guidelines for a comprehensive rodent anesthesia and analgesia program.

OBJECTIVES

When you complete this program, you should be able to: 1. Explain the unique challenges in rodent anesthesia. 2. List the criteria used to select an appropriate anesthetic. 3. List characteristics of individual animals that affect anesthetic choice. 4. Describe the advantages and disadvantages of injectable administration of anesthesia. 5. Describe the advantages and disadvantages of inhalant administration of anesthesia.

LABORATORY ANIMAL MEDICINE AND SCIENCE - SERIES II

1. Series

Laboratory Animal Medicine and Science Series II

2. Title

Rodent Anesthesia & Analgesia

3.

Objectives

When you complete this program, you should be able to: 1. Explain the unique challenges in rodent anesthesia. 2. List the criteria used to select an appropriate anesthetic. 3. List characteristics of individual animals that affect anesthetic choice. 4. Describe the advantages and disadvantages of injectable administration of anesthesia. 5. Describe the advantages and disadvantages of inhalant administration of anesthesia. Rodent anesthesia offers unique challenges that are infrequently encountered in any other veterinary setting. Among the unique challenges in rodent anesthesia is that often surgery is done on more than one animal at a time in an "assembly-line" or "herd" fashion. To accommodate the simultaneous anesthesia of multiple animals, it is useful to have an anesthetic solution available that can be administered based on body weight and that is fairly tolerant of 5, 10, or 20 g weight differences among the patients. This allows parenteral administration of anesthesia on a milliliter per kilogram basis with premixed dilutions or combinations of drugs. There are many anesthetic drugs that have been developed for use in humans or in other animals. Extrapolation of dosages for rodents is sometimes difficult. On the other hand, some anesthetic agents are used only in rodents, but there is very little physiologic, toxicologic, or pathologic information in the literature about their efficacy. In addition, the various species and strains of rodents may show differing sensitivity, efficacy, and duration with various anesthetics. Monitoring physiologic parameters during anesthesia in rodents is infrequent or minimal at best, and because of their small size, it is difficult to use criteria like muscle relaxation and jaw tone to determine the depth of anesthesia, as one does in other species.

4. Introduction

5. Unique Factors

6. Criteria for selecting anesthesia When trying to determine the most appropriate anesthetic for use on rodents, several criteria should be considered: First, what is the purpose of the anesthesia? The purpose will directly influence the duration of anesthesia--whether for short term restraint in order to perform a physical exam or radiograph or for long-term general anesthesia for a complicated protocol.

V-9054 Lab Animal Medicine: Rodent Anesthesia & Analgesia

Second, what is the type of experiment or procedure? Is this a survival procedure or a non-survival procedure? With a survival procedure, it is important that the anesthesia has minimal adverse effects, thereby minimizing its influence on the data being collected. For example, for a respiratory study, opioids which are known to cause respiratory depression should be avoided. Third, is a postoperative analgesia required? Some anesthetics provide better postoperative analgesia than others. Consider also if the appropriate equipment is available. For example, inhalation anesthetics require equipment to administer and an adequate method of scavenging the waste gas so personnel are protected from exposure. In addition, consider both the skill and experience of the anesthetist, because the margin of safety varies among different types of anesthetics. In general, more technical skill is required to administer inhalants compared with injectables. Regulation of the agent should be taken into account since some anesthetics are controlled substances and require a DEA license for purchase. The final criteria to consider are the costs of the agent and the delivery equipment. 7. Patient characteristics

Characteristics of the individual patient that need to be considered when selecting anesthetics include: Species and strain Age, size, and sex Physical condition Temperament Health (preexisting diseases) Previous administration of other drugs Be aware of the health status and physical condition of the patient. Of special importance is any history of previous administration of other drugs or other disease state, which may influence the enzyme systems that metabolize the anesthetic.

8.

Physical restraint Anesthetic induction requires some form of physical restraint. The duration of restraint will vary with the type of anesthetic agent being administered as well as the route of administration. While there are a variety of restraint devices available, it generally requires more time to place the animal into the restraint device than to gently hold the animal during the initial induction phase. The important point to remember is that manual restraint should be brief, in order to minimize stress to the animal. When using inhalant anesthetics, the use of an induction chamber may decrease stress, depending on the species. Also take a look at handling and restraint in V-9042 Rats & Mice: Care and Management , V- 9029 Hamsters: Care and Management, and other rodent programs.

LABORATORY ANIMAL MEDICINE AND SCIENCE - SERIES II

METHODS OF DELIVERY 9. Methods of Delivery This section discusses methods of anesthetic delivery and applicable equipment. In general, anesthesia is administered to rodents by either the parenteral route or the inhalation route. 10. Parenteral route intraperitoneal (IP) intramuscular (IM) intravenous (IV) subcutaneous (SC) Inhalation route open drop delivery precision vaporizer

Disadvantages of Injectables The disadvantages to using injectable anesthetics: Once a given dose is administered, the anesthetic cannot be changed. The duration of anesthetic effect is patient-dependent. There is individual variability for a given dose. Some injectable agents are irritating and measures need to be taken to minimize this effect

11. Dilutions

Prior to administering injectable anesthetics, consider the drug volume, the site, and any irritant properties of the drug. Because concentrated agents are delivered to very small animals, the actual drug volume may be quite small. Many agents can be diluted with physiologic saline to at least a one to one solution (1:1) and, in some cases, one to ten (1:10). Dilutions decrease the chance of overdosing, facilitate drug absorption, minimize irritation, and increase the accuracy of volume measurement by making sure that a significant percentage of the compound is not left within the "dead space" of needle and syringe. If a very large volume of drug needs to be administered, divide the dosage among sites or use more than one route if appropriate.

V-9054 Lab Animal Medicine: Rodent Anesthesia & Analgesia

12. IM Route IM injections should be administered in a large muscle mass, and the two most common sites used in rodents are the thigh muscles and the epaxial muscles located along the spine. Small gauge needles, 26 to 30 gauge, are generally recommended. Large volume doses can be administered in several IM sites and dilutions are recommended if a compound may have irritating properties. IM injections in rodents may be problematic, particularly given the small muscle mass of these animals. Muscle damage may occur from the needle if the animal struggles during the injection, or the injected substance itself may cause damage due to the volume or tissue reaction.

13. IP Route The intraperitoneal route is probably the most common for administering injectable agents in rodents. The technique is relatively easy to master, and many anesthetics which create irritation when given IM or subcutaneously (SC) do not cause lesions or clinical evidence of pain when given IP. In addition, many of the published drug dosages in the literature are formulated for IP delivery. This photo shows the proper site for IP injections in the lower left abdominal quadrant with the animal's head in a down position which allows gravity to move abdominal viscera away from the injection site (drug volumes in needles are for demonstration purposes only). Depending on the size of the animal, a 20 to 23 gauge needle is recommended. It is also important that the needle is long enough to span the thickness of the abdominal wall and actually penetrate into the abdominal space.

LABORATORY ANIMAL MEDICINE AND SCIENCE - SERIES II

14. IV Route Intravenous administration of anesthetics is not commonly used in rodents, because access to peripheral vessels is limited, and the techniques are difficult. In rats, mice, and gerbils, the lateral tail vein is the peripheral vessel most easily accessed. To facilitate access, the vein can be vasodilated by heating, for example, in warm water. In guinea pigs and hamsters, the ear vein, dorsal metatarsal vein, and the pubic or penile vein are the only accessible peripheral vessels. In general though, these require marked restraint and are not practical for induction of anesthesia. However, they could be used to administer compounds in a previously anesthetized animal. 15. Recommendations for IV administration There are some general recommendation for IV administration of anesthetic agents in rodents because the vessels involved are very small. These include: using small gauge needles and small gauge catheters (26 to 30 gauge), injecting materials slowly and stabilizing the needle or catheter once it is in place. After an injection is completed, digital pressure is used to aid in hemostasis. These recommendations are vital to decrease potentially painful sequelae and to preserve vessels if the animal is part of a chronic study.

16. Inhalant anesthetics- advantages Inhalation anesthesia involves delivery of a volatile anesthetic agent to the patient via the respiratory tract. Advantages of using inhalants: Increased control over depth as well as duration of anesthesia. Increased safety and survivability. Minimal metabolism, biotransformation and excretion compared with injectables, which is desirable in a toxicology studies. Animals generally wake up faster from inhalant administration requiring less patient support during the postoperative recovery phase.

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17.

Inhalant anesthetics- disadvantages Most of the disadvantages associated with inhalants involve the required technical skills and the expense of specialty equipment. In addition, an appropriate scavenging system is needed to protect personnel and to comply with OSHA regulations and requirements.

18. Open Drop-bell jar There are two methods of inhalation anesthesia: open drop and precision vaporizer. A transparent bell jar is used in an open drop delivery system with the anesthetic gas placed on cotton balls or sterile gauze at the bottom of the jar and covered with a wire screen mesh. The mesh separates animals from contact with the anesthetic and prevents irritation, and the transparency allows easy observation of the animals. The open drop system does not permit control over the concentration of anesthetic within the jar, therefore only gases with low vapor pressure such as methoxyflurane should be used. The jar must be placed in a well ventilated fume hood whenever it is opened to protect personnel from exposure to the anesthetic gas.

19. Vaporizer If gases with high vapor pressure are used, for example halothane or isoflurane, a calibrated vaporizer must be used to control the concentration of anesthetic. A rodent is placed in an induction chamber connected to the vaporizer. Once the animal is anesthetized, it can be removed from the chamber and maintained on anesthetic gas for the remainder of the procedure. Maintenance is most commonly done by means of a face mask. Face or head masks are commercially available, but are also easily made from a funnel or the proximal end of a 20, 50, or 60 cc syringe barrel depending on the size of the rodent.

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20. Endotracheal intubation Use of a facemask is probably the most common means of delivering anesthetic gas to rodents; however, it is possible to use endotracheal intubation. In rodents, endotracheal intubation is most easily accomplished by direct visualization of pharyngeal and laryngeal structures. This can be accomplished by using a pediatric laryngoscope with a Wisconsin size 0 blade, filed down to make it narrower, and a fiberoptic light or penlight for a light source. Another option is to use a functioning otoscope which serves as both a light source and a speculum and has the added advantage of magnification. Endotracheal tubes can be fabricated using intravenous over-theneedle catheters. Catheter sizes range from 14 to 20 gauge depending on the size of the rodent being intubated, and the tip should be blunted to prevent trauma. Respirators specifically designed for ventilation of rodents are available.

21. Inhalation setup with scavenging system An appropriate scavenging system is a crucial part of administering inhalation anesthetics to rodents. A calibrated vaporizer feeds anesthetic gas into an inhalation chamber. Attached to the opposite side of the inhalation chamber is a facemask, and beside the facemask is a vacuum-scavenge unit. Another option is a carbon-filter gas scavenge canister.

22.

Parallel tube scavenging An alternative scavenging system is to tape two parallel tubes on either side of a funnel facemask. The tubes are connected to a vacuum system and a rodent's nose is placed in the cone. Any escaping gases would be suctioned away by the parallel tubes and protect the surgeon and other personnel in the area from exposure to the anesthetic.

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INJECTABLE ANESTHETIC AGENTS 23. Scavenging options In summary, it is necessary to provide adequate scavenging of the volatile gases when using inhalation anesthesia. The options addressed in this program include use of a fume hood, anesthesia machine scavenging, counter sweeps, and parallel face mask suction.

24. Hypothermia Hypothermia has been described as an adequate anesthetic for short procedures in neonatal rodents. The neonate is encased in a latex sleeve and immersed up to the shoulders in an icewater slurry for 3 to 4 minutes. After the procedure, the neonate is rewarmed in an incubator at about 33 degrees. (Dannemann and Mandrell) 25. Injectable anesthetic agents There are numerous injectable anesthetics available for use in rodents. Some of the more popular agents include: pentobarbital ketamine tribromoethanol

26. Pentobarbital Pentobarbital is a barbiturate and, historically, the most commonly used anesthetic in rodents. An advantage of pentobarbital is that, at recommended doses, it causes minimal cardiovascular depression. It is also relatively long acting and can provide approximately 45 minutes of surgical anesthesia. There are, however, disadvantages to the use of pentobarbital. Pentobarbital is a potent inducer of the hepatic microsomal enzyme system. causes pronounced respiratory depression as well as hypothermia, particularly when repeated doses are given. has prolonged anesthetic recovery which may be accompanied by excitement. is a controlled substance, which requires a DEA license for purchase and appropriate record keeping and storage. is a poor analgesic in many species and can cause high mortality in hamsters and gerbils because the margin of safety is narrow in these species. Even thought the list of disadvantages appears long, by being aware of them, one can successfully use this drug in rodent anesthesia

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27. Tribromoethanol In most rodents, tribromoethanol produces good surgical anesthesia, with good skeletal muscle relaxation and only a moderate degree of respiratory depression. It is relatively inexpensive and not a controlled agent. The major disadvantage of tribromoethanol is a potential for causing peritonitis. When exposed to either light or temperatures >40C, tribromoethanol degrades into two byproducts: hydrobromic acid and dibromoacetaldehyde. Both of these compounds are highly irritating when administered IP and result in peritonitis and visceral adhesions which may be fatal. If prepared correctly, stored at appropriate temperatures, and kept in light-resistant bottles, tribromoethanol can be used successfully ( Kohn). 28. Urethane

Urethane produces long lasting anesthesia with rapid onset following IP administration. It has minimal effects on the cardiovascular and respiratory systems. Urethane provides good analgesia as well as excellent muscle relaxation. Disadvantages to using urethane: At high doses, hypotension, hypothermia, bradycardia, and metabolic acidosis are seen. It is absorbed through the skin and, following topical administration to rodents, can produce sedation and ataxia. It is a proven carcinogen and mutagen in rodents.

If urethane is selected as an anesthetic, personnel must take precautions to adequately protect themselves while mixing a urethane solution. Proper protection should include a double set of gloves, face mask, and mixing under chemical fume hood. Urethane is not recommended for survival procedures.

29. Chloral hydrate Chloral hydrate is a hypnotic and a sedative that, at a high dose (in the range of 450 ml/kg), gives excellent analgesia. Unfortunately, along with this excellent analgesia at high dose, there is uncompensated metabolic acidosis, hypotension, and bradycardia, as well as severe hypothermia. In order to get good analgesia, the dose is such that there is a very narrow margin of safety with a dangerously deep plane of anesthesia and a potential for significant mortality. The abdominal viscera of this rat demonstrates the distension of adynamic ileus, a sequela to a high concentration injection of chloral hydrate administered IP. Chloral hydrate also has the potential for causing peritonitis, however, with dilute concentrations, this complication can be avoided.

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30. Alpha chloralose

Alpha chloralose is another agent that is a hypnotic and a sedative. Historically it has been popularized for its lack of baroreceptor depression which produces a stable, physiologically awake animal that is immobilized and appears anesthetized. A disadvantage of alpha chloralose is that it produces a slow onset of sedation rather than a surgical plane of anesthesia. This sedation is characterized by myoclonia, hyperacusia, persistent muscle tone, convulsion, mild hypothermia, and acidosis. Thus, this drug is probably a hypnotic rather than a true anesthetic and has unproved analgesic potency in rodents. Commonly this agent is used in the second phase of a two-phase procedure. In the first phase, an initial surgery is performed with a potent short-acting anesthetic. In the second phase, alpha chloralose is used to maintain long term immobilization for nonstimulating or nonpainful procedures.

31. Ketamine hydrochloride Ketamine hydrochloride, a dissociative anesthetic, disrupts pain transmission and suppresses spinal cord activity with some action at opioid receptors. Visceral pain is not abolished with dissociative anesthetics, and there is poor muscle relaxation and analgesia. Ketamine is a poor anesthetic when used alone, but is more often combined with other agents. When combined with other drugs, it is usually administered IP. Ketamine is acidic, can be irritating, and cause muscle necrosis when administrated IM. Ketamine-induced nerve damage can cause selfmutilation in rodents. Ketamine is a controlled substance. Store in a locked cabinet and maintain a log of its use. 32. Ketamine + xylazine Drugs often used in combination with ketamine are xylazine, acepromazine, and diazepam. The ketamine/xylazine combination offers superior analgesic activity and is the combination most frequently used. Advantages of ketamine/xylazine include rapid induction with a relatively long duration (60 to 90 min.) of anesthesia plus good analgesia. Upon initial administration, there is transient respiratory and cardiovascular depression. There is also excellent muscle relaxation, and both the induction and recovery phase are smooth. In general, ketamine/xylazine is administered IP in most rodents. The ketamine/xylazine combination, particularly at high doses, can cause significant hypothermia and appropriate measures should be taken to minimize this disadvantage. This combination can also increase intraocular pressure, result in hyperglycemia due to an increase in hepatic glucose production and a decrease in insulin,

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33. Yohimbine

and also decrease gastrointestinal motility.

Yohimbine is a central alpha-2 adrenergic receptor antagonist that can be used to reverse central nervous system (CNS) depression, sedation, bradycardia, and respiratory depression caused by xylazine. It works within 1-3 minutes when administered IV and within 10 minutes when administered IP. Adverse effects of yohimbine are excitement and seizures. Yohimbine is used to allow animals to recover more quickly from the anesthetic combination of ketamine/xylazine. However, in some cases this may not be desirable, and it may be better to let the animals have a slow, gentle recovery.

34.

Ketamine + diazepam Ketamine/diazepam has rapid onset with about 45 to 60 minutes duration of anesthesia. Ketamine/diazepam causes the least respiratory and cardiovascular depression of all ketamine tranquilizer combinations. The disadvantages of using ketamine/diazepam are some muscle rigidity and hyperacusia at low dosages, poor analgesia compared to ketamine/xylazine, and a chance for significant hypothermia. Ketamine/diazepam is selected for studies that require the least respiratory and cardiovascular depression.

35. Tiletamine + zolazepam A drug combination consisting of tiletamine, a dissociative anesthetic, and zolazepam provides anesthesia suitable for restraint, blood sampling, or minor manipulative procedures; however, it provides very poor analgesia. At high enough doses to provide sufficient surgical anesthesia, there are problems with prolonged recovery, severe hypothermia, and hyperacusia. Tiletamine/zolazepam is generally not recommended for use in rodents for surgical anesthesia.

36. Fentanyl + fluanisone The combination of fentanyl and fluanisone is a neuroleptanalgesic. This combination provides reliable surgical anesthesia and analgesia with good muscle relaxation. Non-purposeful limb movements are inhibited. Disadvantages of the fentanyl/fluanisone combination are: mild to moderate respiratory and cardiovascular depression, hypothermia, and difficult to obtain in the United States, though this combination drug is a very popular general rodent anesthesia in Europe. Midazolam, which is often combined with fentanyl/fluanisone, is water soluble, non-irritating, and short acting, and provides additional analgesia.

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37. Propofol Propofol is an hypnotic agent administered IV. Typically rodents are initially anesthetized with another agent, and long term anesthesia is maintained by IV propofol infusion. It is considered a poor analgesic, but offers the advantage of rapid recovery. A disadvantage of propofol is that it requires IV infusion, limiting its use in guinea pigs, hamsters, and gerbils. 38. Local anesthetics Local acting anesthetics, such as lidocaine, or longer-acting bupivicaine, may be infiltrated into tissues as an adjunct to general anesthesia. Long acting local anesthetics may reduce postoperative pain perception, reducing need for general analgesics. Typically, a small volume of local anesthetic is infiltrated into a tissue in the area where an incision will be made (preemptive) or has been made. INHALATION ANESTHETIC AGENTS 39. Inhalant agents The following discussion covers inhalant anesthetic agents which include: halothane isoflurane methoxyflurane carbon dioxide 40. Halothane Halothane is readily available, a potent anesthetic, and not flammable or explosive. It has more rapid induction and recovery compared to ether (generally discouraged as an explosive hazard) and methoxyflurane. Some disadvantages are that halothane: reduces cardiac output. causes peripheral vasodilation leading to hypotension or low blood pressure. is a dose-dependent respiratory depressant. Halothane requires some hepatic metabolism in order to be excreted and has been shown to be hepatotoxic to guinea pigs with repeated use. Halothane must be scavenged as it can cause epinephrine-related cardiac arrhythmias. 41. Isoflurane Isoflurane is nonflammable and nonexplosive. Compared to halothane, isoflurane causes less depression of cardiopulmonary function. less sensitization of the heart to catecholamine (beta-adrenoceptor agonist) release. less profound respiratory depressant effect. Note that isoflurane reduces renal blood flow, glomerular filtration rate and urinary flow. It is not a significant hepatotoxin. Isoflurane's metabolism to organic and inorganic flourides is less than any other halogenated agent available, so if a minimally metabolized anesthetic is

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needed, isoflurane is the choice. Both induction and recovery from isoflurane are rapid.

42. Methoxyflurane

Methoxyflurane has recently become unavailable in the United States, but is covered here since it is still available elsewhere. Methoxyflurane is nonflammable, nonexplosive, and a potent analgesic. Low vapor pressure makes it the only agent suitable for the open drop method. A disadvantage of methoxyflurane, compared to other inhalents, is a relatively slow induction phase that can result in a modest respiratory and cardiovascular depression. Perhaps the biggest disadvantage is that metabolization leads to flouride ion release which is nephrotoxic. An appropriate scavaging system must be in place to protect personnel. The utility and humane acceptability of CO2 and O2 combination as an anesthetic is an area of contention. This combination is used for extremely short term procedures. Some studies have found that low concentrations (50% CO2) lead to prolonged induction and severe and frequent adverse effects that include nasal bleeding, excessive salivation, seizures, and even death. Humans, when asked to voluntarily breath 50% CO2, rated the experience as unpleasant. High CO2 concentrations (100%) produce rapid anesthesia with few adverse effects, but when humans were asked to breathe 100% CO2, they reported that this experience was extremely painful. A recommendation is to provide CO2 in the ratio of 70% CO2 to 30% O2 for anesthesia, as a compromise of humane acceptability and practicality (Kohn).

43. Carbon dioxide

MONITORING 44. Anesthetic depth Throughout the course of anesthesia, animals should be observed for pink mucous membrane color, a general indication of aqequate oxygenation. Respiratory rate should remain even. Movement or change in respiratory rate or depth in response to manipulation may indicate insufficient depth of anesthesia. Confirm the animal's lack of response to stimulation, such as withdrawal from a paw pinch, every 15 minutes or so throughout a surgical procedure. Pulse oximetry can be used as an objective measure of oxygen saturation and pulse rate. Several units are available that can be used with rodents such as rats. However, small size and lack of a tail in some species limit sites for sensor application.

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ANALGESIA 45. Assessment of pain Pain in rodents is most commonly assessed on the basis of subjective measures and observations combined with professional judgment. Clinical assessments of pain include behavioral changes such as immobility, unkempt appearance, lack of appetite, abnormal vocalization, and abnormal posture. Animals experiencing pain may appear unresponsive lethargic anxious apprehensive hypersensitive aggressive Any single measure is not pathognomonic for pain. Porphyrin tear staining may be noted in rats. Animals may chew on themselves or pull hair. Physiologic signs such as pupil dilatation, fluctuations in blood pressure, increased heart rate, respiratory rate, and body temperature may be more difficult to routinely assess. Implanted telemetry devices are used in some situations to provide objective data on some of these parameters. Changes in weight can be readily measured. When pain or distress is suspected, humane considerations dictate that relief be provided to an animal.

46.

Herd analgesia Rather than assessing individual animals for signs of discomfort, investigators may prospectively design an analgesic protocol which will make postoperative analgesic therapy available to each study participant. The strategies involve recognizing and eliminating pain in groups of rodents, rather like a herd approach to analgesia. In this approach, the pain intensity of a given research procedure, based upon known discomfort levels in larger companion animal species or humans, should be considered. The prediction of pain intensity is often based on the type of procedure and the location of the lesion. (Kohn)

47.

Salicylates Acetylsalicylic acid (aspirin) and sodium salicylate are used to relieve only mild to moderate pain; they do not relieve deepseated visceral pain or sharp intense pain. Aspirin is not routinely administered to rodents because of the difficulty in dissolving the drug in water or other oral solutions.

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48. Morphine-sulfate Morphine-sulfate is a potent narcotic analgesic. It is a controlled drug and must be stored in a locked cabinet and a log of use maintained. Investigators need to be aware that animals will develop tolerance to morphine with chronic administration. Morphine has a short duration of action and must be readministered at frequent intervals usually of less than 4 hours. Morphine can induce respiratory depression and sedation. 49. Meperidine Meperidine is a narcotic sedative and analgesic that is less potent than morphine. It can be used as a postoperative analgesic and its respiratory depressant effects are similar to morphine. An advantage is the effects can be reversed by an antagonist. Effects of meperidine are short lived, usually about 4 hours. Like morphine, meperidine is a controlled drug. Since it is not as potent as morphine, there is no particular advantage to using it instead of morphine when a potent, short-acting analgesic is indicated. 50. Buprenorphine Buprenorphine is an opioid with prolonged duration of action, ranging from 6 to 12 hours. High dosages can negate the analgesic effect, and complications from repeat high dosage include hematuria and GI bleeding. (This photo shows hemorrhage in the bladder of a rat given repeat high dose buprenorphine.) At lower doses buprenorphine is an effective analgesic in rodents. (Liles, Flecknell, Roughan, and Cruz-Madorran)

51. Other analgesics Use of other analgesics has been described. Analgesics with less utility in rodents include pentazocine, butorphanol, flunixin meglumine, and nalbuphine. Pentazocine lactate is a nonnarcotic analgesic used in postsurgical care. The respiratory depressant effects of pentazocine are considered less than those of meperidine, but unfortunately, it does not work well in rats. Butorphanol tartrate is a synthetic analgesic with narcotic agonist/antagonist properties. In humans, it is three to five times more potent than morphine. Unfortunately, it lacks potency in rodents. Flunixin meglumine is an agent that is used commonly in humans and dogs. It is a potent non-narcotic, nonsteroidal analgesic agent with anti-inflammatory and antipyretic activity. Again, it is not useful in rodents-often resulting in a hyperexcitable animal.

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Nalbuphine is a mixed synthetic narcotic agonist/antagonist analgesic related to oxymorphone and naloxone. However, its very short duration time of minutes in rodents makes it useless in rodent analgesia.

52. Analgesics -preemptive Preemptive analgesia is the administration of analgesic agents prior to or at the time of anesthesia, before any tissue-damaging procedures, such as incisions, have been started. Preempting pain signals in this way have been shown, in humans, to decrease levels of postoperative pain and need for analgesia. Preemptive administration of analgesics such as buprenorphine has also been shown to decrease the amount of isoflurane gas anesthetic needed to maintain a surgical plane of anesthesia in rats (Criado).

53. Analgesics -oral There is no assurance that oral formulations of analgesics, regardless of flavor or formulation chosen, will be voluntarily ingested by a given rodent in sufficient quantity to provide effective analgesia. However, administration of oral buprenorphine mixed with flavored gelatin has been described as an effective analgesic in rats. The rats are preconditioned to take flavored gelatin as a treat. (Flecknell, Roughan and Stewart ) For doses, refer to the Formulary for laboratory animals (Hawk and Leary), Anesthesia and analgesia in laboratory animals (Kohn, Wixson, White and Benson), or Research animal anesthesia, analgesia and surgery (Smith and Swindle).

54. Supportive care Intra- and post-surgical supportive care includes ocular lubrication, topical antibiotics, nutritional considerations, and minimizing heat loss. Rodents typically are not fasted or water restricted prior to anesthesia, as is done with larger species. Typical postoperative nutritional support can include supplemental fluids to maintain fluid volume. One may also provide warmed 5% dextrose, given SC gelatin or agar diets, as well as fruit or vegetables or peanut butter. Use of fresh fruit or vegetables may be restricted in some facilities. Ocular lubrication during the entire anesthetic episode, as well as into the postoperative phase is recommended Antibiotics should be used when appropriate, including the use of topical antibiotics along implant

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exit sites. Antibiotic use should not be a routine antidote for poor intraoperative sterile technique. Care should be taken to minimize heat loss to the environment. This can be done by using a heated OR table, heat blankets, heat lamps, drapes, and minimizing organ exposure from body cavities. While animals are recovering from anesthesia, rodents may huddle with cage-mates for warmth. However, anesthetized rodents should not be caged with awake animals. Return each animal to group housing only when confidant the animal is able to move about and defend itself. The V-9055 Rodent Surgery program contains a thorough discussion of supportive care.

54. ACLAM Credits The Laboratory Animal Medicine and Science _ Series II has been developed under the following committee for the American College of Laboratory Animal Medicine C. W. McPherson, DVM, Chair J. E. Harkness, DVM J. F. Harwell, Jr., DVM J. M. Linn, DVM A. F. Moreland, DVM G. L. Van Hoosier, Jr., DVM L. Dahm, MS. 59. HSCER Credits Produced by the Health Sciences Center for Educational Resources, University of Washington, Seattle WA 98195 2000

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References American Veterinary Medical Association. 1993. Report of the AVMA panel on euthanasia. J. Am. Vet. Med. Assoc. 202(2):229-249. Canadian Council on Animal Care . 1993. Guide to the care and use of experimental animals, vol. 1. Canadian Council on Animal Care, Ottawa, Ontario. [http://www.ccac.ca/] Criado, A. B., A. Gomez de Segura, F. J. Tendillo, and F. Marisco. 2000. Reduction of isoflurane MAC with buprenorphine and morphine in rats. Lab. Anim., 34: 252-259. Danneman, P. and Mandrell, T. 1997. Evaluation of Five Agents/Methods for Anesthesia of Neonatal Rats. Laboratory Animal Science, August (47) 4. Flecknell, P. 1996. Laboratory animal anesthesia, 2nd ed. Academic Press, Inc., San Diego. Flecknell, P. A., J. Roughan, and R. Stewart. 1999. Use of oral buprenorphine ("Buprenorphine jello") for post-operative analgesia in rats - a clinical trial. Lab. Anim. 33, 169-174. Hawk, C., and S. Leary. 1999. Formulary for laboratory animals, 2nd ed. Iowa State University Press, Ames. Jenkins, W. L. 1989. Pharmacologic aspects of analgesic drugs in animals: an overview. J. Am. Vet. Med. Assoc. 191(10):1231-1240. Kohn, D., S. Wixson, W. White, and C. Benson. 1997. Anesthesia and analgesia in laboratory animals. ACLAM, Academic Press, New York. Liles, J. H., P. A. Flecknell, J. V. Roughan, and I. Cruz-Madorran. 1998. Influence of oral buprenorphine, oral naltrexone or morphine on the effects of laparotomy in the rat. Lab. Anim. 32, 149-161. National Institutes of Health. 1996. Guide for the care and use of laboratory animals, p.48. NIH publication No. 86-23. National Institutes of Health, Bethesda, Md. Smith, A., and M. Swindle. 1994. Research animal anesthesia, analgesia and surgery. Scientists Center for Animal Welfare, Greenbelt, Md. U.S. Department of Agriculture. 1966. Animal Welfare Act, (P.L. 89-544). Amended 1970 (PL 91-579); 1976 (PL 94-279); 1985 (PL 99-198). Animal and Plant Health Inspection Service, U.S. Department of Agriculture, Hyattsville, Md.