Name ANIMAL SCIENCE 140 LABORATORY ANIMAL MANAGEMENT MIDTERM I Matching (20 points) Choose the most correct answer identified by the statements given. Each answer may be used more than once. There are 2 sections of matching questions. Use only the answers applicable to the section you are working on. A. Mouse C. Hamster E. Guinea Pig B. Rat D. Gerbil F. Rabbit 1. 2. 3. 4. 5. 6. 7. 8. 9. Sexing of this species takes practice because both the male and female have nipples and there is no difference in the anal-genital distance. You must rely on the Y versus rosette shapes. A very short tail, lots of loose skin, and an aggressive nature makes this the most difficult lab rodent to handle. This species doesn t have the longest gestation, but at 5 to 6 months, it definitely is oldest when reaching puberty. This predominately diurnal, desert rodent is a favorite in labs outside of California because of its friendly nature and clean cage habits. This species can be used to study hibernation and to look at implanted tumor cells growing in vivo. Of the altricial rodents, this species has the longest cycle (6 days) and the longest gestation (25-26 days). All rodents have a Hardarian gland; however, in this species, stress causes an overproduction of porphyrin resulting in red tears. Mus musculus This species has a narrow palatal ostium to prevent aspiration of food if it is startled and begins to stampede. You shouldn t gavage them. 10. In this species, males are likely to fight and can t be regrouped after puberty, but females are tolerant of other females and can be mixed. 1
A. Animal Welfare Act (AWA) B. Federal Food, Drug, and Cosmetic Act C. Good Laboratory Practices Act (GLP) D. Health Research Extension Act (HRE) E. None of the above 11. This act covers certain mammals, including lab-bred hamsters and wild-caught mice, but generally excluding domestic mice and rats. 12. This was enacted following the death of 107 children given an elixir which used antifreeze as the vehicle. 13. This act uses the Institute of Laboratory Animal Resources (ILAR) Guide as its set of rules. 14. This outlines a strict, unbiased set of rules that must be followed when testing the safety of any drugs intended for human or animal use. 15. Either one of 2 acts that mandate the formation of an Institutional Animal Care and Use Committee (IACUC) to oversee animal research. (Yes, there are 2 possible answers to this question. Either will be accepted.) 16. This act requires safety testing using live vertebrate animals for all new cosmetic formulations in the U.S. 17. This act covers all vertebrate species used in research or teaching in an institution receiving any government funding. 18. This act is overseen by the National Institute of Health (NIH) and certification of compliance is granted following a peer-review inspection by members of the Association for the Accreditation and Assessment of Laboratory Animal Care (AAALAC). 19. This act covers livestock used as food or fiber. 20. This act mandates regular inspection of facilities housing covered species by United States Department of Agriculture (USDA) veterinarians. 2
Multiple Choice (50 points) Please circle all correct answers (from 1 to all 5 may be correct). 21. Frequent injections can be irritating causing both stress from handling and also scar tissue. If you need to give a specific dose of a drug over a long period of time, which of the following are the best options. a) Increasing the dose of each injection so that the effects last longer and fewer injections need to be made. b) Implanting a catheter so a drug can be administered without an additional restraint or needle stick. c) Varying your routes from subcutaneous to intraperitoneal to intradermal so that there is less build-up of scar tissue. d) Implanting an osmotic pump in the subcutaneous space to deliver the drug continuously over time. e) Putting the drug into the animal s drinking water. 22. When planning a blood collection a) Choose the smallest gauge needle that will penetrate the skin. b) Choose a receptacle that will not draw too strong a vacuum. c) Heating pads may be used to increase vasodilatation. d) Always use a topical analgesic. e) Always collect from a vein; never an artery. 23. When using ketamine as an anesthesia for a rat, you should know the following: a) Ketamine is usually combined with a sedative thus lowering the required dose and decreasing the potential side effects. b) Ketamine is a good muscle relaxant which reduces pain and trauma during surgery. c) You can monitor anesthetic depth and determine whether the animal has reached the surgical plane by looking for the loss of the toe pinch reflex. d) The eye blink reflex can be used to determine when anesthesia levels are becoming too light and the animal needs to be re-dosed. e) You are trying to anesthetize a 100 g rat. Your formulary dose is 100 mg/kg. Your drug concentration is 100 mg/ml. You calculate your delivery dose as follows: 100 mg/kg x 1 ml/100 mg x 0.1 kg = 0.1 ml 24. Cardiac puncture as a method of blood collection a) It can be done on rodents but not rabbits b) It requires anesthesia when performed on a live animal c) It can be done immediately following euthanasia d) It is a terminal procedure in small animals e) It requires a surgical cut-down to expose the xiphoid process 3
25. An intraperitoneal (IP) injection a) It is a route used for all rodents but not the rabbit b) It has a slower absorption rate than a subcutaneous injection. c) It requires aspiration to ascertain needle placement. If you see blood in the syringe, throw the syringe out and start over. d) Ir is given on the right side of the abdomen to avoid insertion into the bladder or cecum. e) It is an appropriate route to administer a euthanasia solution to small animals as long as the chemical is not irritating. 26. Buprenorphine is an analgesic commonly given to laboratory animals. a) It is a very powerful analgesic and is a level 1 controlled substance. b) It is long lasting, often giving 8 to 12 hours of pain relief. c) It causes a decrease in gastric motility so animals may not feel like eating and lose weight. d) Like most analgesics, it is most effective if the first dose is given immediately after the animal wakes up from anesthesia. e) It is usually administered as a subcutaneous injection, but it may be offered in strawberry gelatin for relief of mild pain. 27. Which of the following subjects are addressed in the protocol for IACUC review? a) Reduction of study duplication through adequate database research. b) Justification of species and animal number to assure that the research will be accurate and the data statistically significant. c) Reduction of animal discomfort by addressing adverse effects. d) Assurance of appropriate veterinary care. e) Selection of euthanasia appropriate for the species and age of the animal according to the AVMA. 28. You are preparing a rat for a vasectomy. a) Clip all the fur from the belly of the rat. b) Use a surgical scrub (detergent/disinfectant) to clean the skin. c) Scrub in a back and forth pattern to make sure the skin is clean all the way to the edges. d) Rinse between scrubs using water, saline, or alcohol. e) After 3 scrub-rinse cycles, finish with the surgical scrub leaving the detergent on the skin for a residual disinfectant effect. 4
29. You are working on your Table of Study Groups for an experiment that involves feeding pregnant rats a suspected toxic substance to see if the offspring are affected. Your study design requires mating 6 experimental dams and 6 control dams to a group of males. Each dam will give birth to 1 litter with an expected average litter size of 10 pups per litter. You plan to euthanize 2 pups per litter at each of the following time points: birth, 8 days of age, and 21 days of age. How many groups are required? a) 2 b) 5 c) 9 d) 11 e) 12 30. If pain is expected in your experimental design, the adverse effects section of your protocol may include a statement like, Analgesics will be administered every 8 hours for 24 hours. Pain relief will be monitored and additional analgesics administered if any of the following signs are observed. a) Hunched posture b) Sunken eyes c) A body condition score of 5 d) A body condition score of 2 e) A body condition score of 1 5
Essay Questions (30 points total) 31. You are doing a pharmacological study on rats that involves oral administration of a fat-soluble drug followed by serial blood draws every 30 minutes for 5 hours (10 total blood collections) to look at absorption, distribution, and elimination of the drug. (10 points) a) Which method of administration would you use and why? b) Which method of blood collection would you use and why? c) How would you express your maximum collection volume in your protocol where you will have rats with different weights and what would be the maximum volume you could collect from a 200 g rat at any time point? d) Briefly describe your control group (what is different between their treatment and that of the experimental group). 6
32. Compare induction and recovery between inhalant and injectable anesthesia. Which class would be more risky for a rat on an alcoholism study with liver damage? (10 points) 7
33. For each of the following scenarios, give your euthanasia choice and explain why you would choose that method as well as any special procedures or precautions. (10 points) a) A study has ended and you need to euthanize 100 adult mice. b) A dam has died and you need to euthanize a litter of newborn rat pups. c) You are writing a protocol for a catch and release field study using live traps for mice. Which method of euthanasia would you list in case a mouse is injured? 8
Answers to Matching and Multiple Choice 1. E 2. C 3. F 4. D 5. C 6. D 7. B 8. A 9. E 10. A 21. bd 22. bc 23. ace 24. bcd 25. ade 26. be 27. abcde 28. bd 29. d 30. abd 11. A 12. B 13. D 14. C 15. A or D 16. E 17. D 18. D 19. E 20. A 9
Essay Questions (30 points total) 34. You are doing a pharmacological study on rats that involves oral administration of a fat-soluble drug followed by serial blood draws every 30 minutes for 5 hours (10 total blood collections) to look at absorption, distribution, and elimination of the drug. (10 points) a) Which method of administration would you use and why? Administration is oral so you will need to either gavage, feed it using an eye dropper, or put it into the food or water. Since you are testing absorption, you need to deliver all at once so putting it into the feed or water won t work. It is hard to dose accurately using an eyedropper. Gavage is the best option. b) Which method of blood collection would you use and why? You need a method which allows for multiple small collections. Lateral saphenous is often used for this type of study. It doesn t require anesthesia and restraint is relatively easy. You can alternate legs and even scrape off the scab for multiple collections without additional punctures. Submandibular collections are also a possibility. It is fast and requires only a good scruff for restraint. However, it is more stressful and not often used for rapid serial collections. The tail vein can be used, but it is more difficult and requires a longer and more confining restraint. c) How would you express your maximum collection volume in your protocol where you will have rats with different weights and what would be the maximum volume you could collect from a 200 g rat at any time point? In the protocol, you just need to write that you will collect a maximum of 1% body weight every 2 weeks. The actual collection volume per animal depends on weight. The collection volume for a 200 g rats is o 0.01 x 200 = 2 ml every 2 weeks o 2 ml/10 blood collections = 0.2 ml/draw d) Briefly describe your control group (what is different between their treatment and that of the experimental group). The control group needs to be gavaged using the appropriate vehicle. Since the drug is fat soluble, corn oil should work. The blood collections will be the same as for the experimental group. 10
35. Compare induction and recovery between inhalant and injectable anesthesia. Which class would be more risky for a rat on an alcoholism study with liver damage? (10 points) Induction of an injectable is via the blood stream. The drug is generally administered in a single bolus which causes a peak as the drug is absorbed and a valley as it is broken down. Re-dosing causes additional peaks and valleys. Induction of an inhalant is via the lungs and is more likely to be administered continuously throughout the procedure resulting in a steady plane of anesthesia. Elimination of injectables requires metabolism (actually catabolism or breaking down of the chemical) and excretion via the urine or feces. This process takes time during which the animal is exposed to side effects such as respiratory depression. It also has the potential to cause damage to the tissues of the liver and kidney where catabolism occurs. Recovery of inhalants is primarily via the lungs. Most, or all, of the drug is blown off intact so liver and kidney involvement is minimal resulting is minimal damage and recovery time is substantially shortened so there is less time exposed to side effects. The injectable is more risky to the animal with liver damage because the liver may not be able to break down the chemical effectively resulting in prolonged exposure to side effects and possible overdose. 11
36. For each of the following scenarios, give your euthanasia choice and explain why you would choose that method as well as any special procedures or precautions. (10 points) a) A study has ended and you need to euthanize 100 adult mice. CO 2 is the euthanasia of choice for large numbers of rodents. It is easier on technicians than injectables which require individual restraint. Requires an individual exam (toe pinch) and/or a secondary method of euthanasia to assure death. A 30% fill rate is required by the AVMA to cause unconsciousness and analgesia prior to feelings of breathlessness, fear, and the carbonic acid burning sensation. CO 2 doesn t poison the carcass so the carcass can be used as food for raptors and reptiles. b) A dam has died and you need to euthanize a litter of newborn rat pups. Altricial neonates resist hypoxia so CO 2 can take 30 to 60 minutes. Freezing is the method of choice for the first 5 days of life. Don t put neonates on a frozen surface during induction. c) You are writing a protocol for a catch and release field study using live traps for mice. Which method of euthanasia would you list in case a mouse is injured? Cervical dislocation is the method of choice It requires no special equipment so can be used easily in the field AVMA grants provisional approval for special circumstances provided the technician is well-trained. 12