Contemporary Topics 36 (3)
1997

The Moribund State as an Experimental Endpoint. Contemporary Topics 36 (3): 44.
This article is a prespective on defining the moribund state in the context of specific exprimental paradigms by the use of objective data-based approaches to predict when an animal is likely to die and thereby could lead to reduced pain, disstress, and suffering in research animals. An approach that can be taken is to focus on specific physiologic or clinical variables (such as possibly body temperature responses or weight loss patterns) identitified in the context of specific experimental paradigms. This data can be collected during initial or pilot studies. This could potentially facilitate the implementation of timely euthanasia prior to the onset of clinically overt signs of being moribund.
No questions

Animal Welfare and Toxicology/Safety Studies: Making Sense of the Regulatory Environment. Contemporary Topics 36 (3): 49.
Animal safety and toxicology studies are used to determine the basis of pharmacological and toxicological profiles, and to predict the hazard potential of a test article. The word nonclinical is preferable to preclinical because animal studies are conducted prior to and during clinical studies. The impact of transgenic animals on animal welfare issues mentioned. Fundamental concepts identified as the decreased use of animals in safety testing, definition of expectations through establishment of endpoints, the sophistication of animal rights activism and how it has integrated itself into mainstream public sentiment, the need for effective leadership in institutions. Economic concerns include animal facility expansion, which many institutions are currently refraining from due to decreased animal use and animal facility management. Management should emphasize that the investigator is the customer and should operate as an extension of the institution's science, safety testing, and collaboration effort.
Review of applicable regulations and guidelines:
1. Food and Drug Administration (FDA), CFR 21 Part 58 (Good Laboratory Practice Regulations). FDA is division of DHHS. Under the Federal Food, Drug and Cosmetic Act, the FDA regulates food and color additives, animal food additives, human and animal drugs, medical devices, biologics, and electronic products. Clinical studies divided into 3 phases:
a) Phase 1- initial short term testing on small group of health human patients
b) Phase 2 - larger study to determine safety and efficacy in target population, ascertain short-term adverse effects and minimum effective dose
c) Phase 3 - expanded study determines further safety and efficacy information to determine risk/benefit ratio
2. EPA - Environmental Protection Agency - oversees use of pesticides and industrial chemicals via Office of Toxic Substances and Office of Pesticide Programs CFR 40, Part 158 - FIFRA - Federal Insecticide, Fungicide, and Rodenticide Act CFR 40 Part 772 - TSCA - Toxic Substances Control Act
3. CPSC - Consumer Product and Safety Commission, CFR 16 Part 1500 Personal care products and household products. Use ingredients referred to as generally-recognized-as-safe (GRAS). Federal Hazardous Substances Act describes label wording that is based on animal safety studies.
4. OSHA - Occupational Safety and Health Administration - CFR 29 Part 1900, Oversees safety in workplace. Requires material safety data sheet (MSDS) for any chemical that a worker may be exposed to. MSDS includes chemical and physical properties, shipping and handling requirements, health hazards, emergency first aid procedures, relevant regulatory information.
Involuntary Animal Welfare Requirements:
1. AWA Pub.L. 99-198, CFR 9 Parts 1,2, and 3
1st passed in 1966 (Lab Animal Welfare Act), amended 3 times AWA administered by USDA through REAC division of APHIS
2. PHS Policy on Humane Care and Use of Laboratory Animals initiated in 1971, revised in 1985 - Health Research Extension Act, Pub.L. 99-158 OPRR of NIH responsible for administration and coordination of PHS policy. Letter of Assurance required by institutions. They must use The Guide as their basis for program development.
3. U.S. Govt. Principles - endorsed by PHS and developed by the Interagency Research Animal Committee (IRAC).
4. GLP Act CFR21 Part 58 - effective 1979; nonclinical safety assessment studies must follow. Includes requirement for creation of SOPs for all aspects of the study and a quality assurance unit, which includes internal inspections of practices and records.
5. Applicable state and local animal welfare laws
Voluntary Animal Welfare Requirements:
1. AAALACI - Assoc. for Assessment and Accreditation of Laboratory Animal Care International; original charter created in 1965; Board of Trustees composed of representatives from 40 professional organizations.
2. The Guide for the Care and Use of Laboratory Animals
1st published in 1963 - developed by the Animal Care Panel subsequent revisions by ILAR. 1996 = 7th edition
Important organizations in industry:
1. PhRMA - Pharmaceutical Research Manufacturers of America
2. BIO - Biotechnology Industry Organization
3. HIMA - Health Industry Manufacturers Assoc.
4. CTFA - Cosmetics, Toiletries, and Fragrance Assoc.
5. CMA - Chemical Manufacturers Assoc.
6. ACPA - America Crop Protection Assoc.
7. Contract labs
Current issues and trends include:
1. the new Guide's emphasis on performance standards/goals
2. the memo of understanding between USDA-APHIS, DHHS-FDA and DHHS-NIH, renewed in 1995
3. the occupation health and safety report under review by ILAR
4. USDA-APHIS-REAC - performance based standards review, team-based organization, updated policy manual, increase injunctive authority being sought to search properties for evidence, and draft proposal to eliminate Class B dealers for research. To AWA amendments currently under proposal include H.R. 3393 the Family Pet Protection Act of 1996, and H.R. 3398 - The Pet Safety and Protection Act of 1996.
5. ICCVAM draft report in conjunction with the National Toxicology Program Workshop - recommends criteria and processes for the validation and regulatory acceptance of new and revised toxicological testing methods that encourage the 3 R's of animal use alternatives.
No questions

A Comparison of Ketamine, Ketamine-Acepromazine, and Tiletamine-zolazepam on Various Hematologic Parameters in Rhesus Monkeys (Macaca mulatta). Contemporary Topics 36 (3): 55.
A comparison was made of the complete blood count, immunohematologic, and serum biochemical values in 18 rhesus monkeys that were immobilized with ketamine, ketamine-acepromazine, or tiletamine-zolazepam (Telazol). Significant differences were not found among the hematologic variables evaluated while the monkeys were under anesthesia induced by use of the 3 anesthetic agents, except for serum potassium concentrations. Although the potassium values were significantly different, values remained within the established normal range of the reference laboratory for rhesus macaques (Macaca mulatta). Tiletamine-zolazpam did not significantly alter the complete blood count, immunohematologic, and serum biochemical values. It may be preferable, because it provides increased analgesia in nonhuman primates, compared with ketamine or ketamine/acepromazine.
QUESTIONS
1) Assuming the null hypothesis is true, the "p" value is the probability of getting results at least as extreme as those in the data. True or False?
2) Telazol combines the positive aspects of ketamine or ketamine/acepromazine in the nonhuman primate with the additional benefit of providing for :
a) maintaining the pharyngeal reflex
b) decreased salivation
c) mild to moderate analgesia
d) no effect on the CBC, immunohematologic, or serum biochemical values
3) Telazol is a combination of a dissociative anesthetic and a nonphenothiazine diazepinone. True or False?
ANSWERS
1) True. The null hypothesis is a statement that we wish to evaluate by examining data. Often the null hypothesis is that a treatment has no effect or is no better or worse than a standard treatment. The "p" value is the probability of getting results at least as extreme as those in the data, assuming the null hypothesis is true. JAVMA, 1990 Aug: 197(3): ?
2) C
3) True

Analgesic Activity of Acetaminophen and Bupreorphine Administered in the Drinking Water of Rats. Contemporary Topics 36 (3): 58.
Authors wanted to determine efficacy of the two analgesics, buprenorphine and acetaminophen, given in the drinking water. They used thermal stimulus to measure the paw withdrawal latency time before and after parenterally administered analgesics or analgesics administered in the drinking water. Giving analgesics in the drinking water is desirable since administration of individual doses can be time consuming, particularly when large numbers of rodents must be treated. The results show that buprenorphine administered in the drinking water at 2.9 mg/kg/day resulted in a significant increase in paw withdrawal latency time but not at lower doses. Acetaminophen administered in the drinking water at up to 600 mg/kg/day did not cause significant differences in paw withdrawal latency time. There is some question whether analgesic doses derived from analgesiometric assays (paw withdrawal assays) are predictive of clinical doses required to produce analgesia and that a real-life stimulus such as surgery or injection of an irritant and then monitoring of a response might more accurately predict a clinical dose. The acetaminophen dose could not be increased because of the risk of producing hepatotoxicosis and nephrotoxicosis when administered long-term to rats. A negative aspect to the use of buprenorphine is its considerable cost. The data in this study need to be further confirmed that results were attributable to buprenorphine drug effect rather than some other effect, as the increases in latency times measured were not significantly different from the changes in the latency times measured in the control groups.
Questions:
1. High doses of acetaminophen will cause what two types of toxicoses?
Answers:
1. Hepatotoxicosis and nephrotoxicosis.

Synthetic Cast Protective Dressing for Skin Experiments in Rats. Contemporary Topics 36 (3): 63.
The use of rats in studies of dermatology or plastic and reconstructive surgery presents a challenge, as a rat can rapidly damage the fragile surgical site. The studies in this paper required protection of the surgical site for 2 weeks. Elastic bandages and plaster casts had several drawbacks, including partial disintegration when wet, respiration depression, interference with movement, eating and drinking. Therefore; the investigators adapted a synthetic cast (ZIMMER CAST-II, Bristol-Myers Squibb Co., Tokyo, Japan) for their studies. A package of cast material was divided for use among 3 rats. The cast was wrapped 3 times around the body of each rat, extending from the forelimbs to the hips of the rat. The large area of coverage was used for 2 reasons: First, to protect the wound from contamination. Second, to increase the security of the cast by allowing it to adhere to the hair. The average weight of the cast was 15 g. This cast had several advantages over the previously mentioned materials. The material was resistant to biting, licking, scratching, and wetting. The cast material was easier to apply and light weight. Animals were able to eat, drink, and move. The cast was easily removed with a pair of scissors. The only disadvantage was the development of sores due to abrasion of the skin by the cast. The authors recommended trimming the cast material or using a flexible cast material (Soft Cast, 3M Health Care, St. Paul, and MN) to avoid abrasions. Additionally, it is important to use the material quickly after the package is opened as the adhesiveness decreases with time and may result in cast failure.
No questions

Breeding Nine-Banded Armadillos (Dasypus novemcinctus) in Captivity. Contemporary Topics 36 (3): 66.
Xenarthra: Dasypodidae are the natural hosts for Trypanosoma cruzi, Toxoplasma gondii, Sarcocystis and Eimeria species, a piroplasm, Leptospira species, Salmonella sp., and Mycobacterium leprae. The armadillo is of interest to the geneticists because they reproduce by polyembroyony to produce 4 (D. novemcinctus to 12 D. hykridus) identical offspring. The armadillo is a source of antigen to prepare M. leprae/BCG mixures for immunotherapy of humans with hanseniosis (Hanson's disease).
The major obstacle severely limiting the use of armadillos as laboratory animals is breeding them in sufficient numbers.
This paper reports the preliminary results of methods that led to success in adapting Dasypus novemcintus to captivity and resulted in the production and survival of two consecutive litters from one captive pair of armadillos. The authors walled off an area of forest for the breeding pair that offered a similar environment to the armadillo's natural habitat.
Questions:
1) Which one of the following organisms is NOT naturally found in the armadillo?
A) Salmonella sp
B) Toxoplasma gondii
C) Sarcocystis sp.
D) Gryllus sp.
E) Salmonella sp
2) Armadillos (edentates) are unique mammals that have been used extensively in leprosy and reproductive studies. True or False
Answers:
1) D
2) True. Edentates are a group of mammals having only molars or no teeth at all, as sloths, armadillos, and anteaters.

Evaluation of Objects for Environmental Enrichment of Mice. Contemporary Topics 36 (3): 69.
This study examines several potential enrichment objects (namely marbles, polyvinyl chloride tunnels, and cellulose nestling material in bedded cages) and compares their use (by inbred DBA/2, outbred CD-1, and crossbred B6CBF1 mice) with control bedded cages. Activity and Inactivity time periods were calculated by videotaping during 6 hour dark cycle using infrared lighting.
The singlely housed mice preferred tunnels and nestlings to marbles. DBA/2 mice were more active than the others. Total activity time was not increased by the enrichment devices but decreased digging activity was noted.
QUESTIONS:
1. All mice exhibited behaviors including eating, grooming, digging, chewing the grommet of the water valve, and exploring. List the activities from greatest to least time involved in the activity.
2. True or False Singlely housed mice spent greater amounts of time playing with their marbles than the nestlets or polyvinyl chloride tunnels.
3. Inbred mice are generally believed to be less physically active than outbred or hybrid mice. Which mice were most active?
4. True or False The Animal Welfare Regulations requires Environmental Enrichment for mice of the Genus Mus.
ANSWERS:
1. All mice exhibited behaviors including eating, grooming, digging, chewing the grommet of the water valve, and exploring. List the activities from greatest to least time involved in the activity.
Grooming - 77.60 min.
Digging - 36.15 min.
Chewing the Grommet - 23.42 min.
Eating - 23.33 min.
Exploring - 21.88 min.
2. True or False Singlely housed mice spent greater amounts of time playing with their marbles than the nestlets or polyvinyl chloride tunnels.
False
3. Inbred mice are generally believed to be less physically active than outbred or hybrid mice. Which mice were most active?
DBA/2 - the inbred mice Moral: Don't always
believe in the general.
4. True or False The Animal Welfare Regulations requires Environmental
Enrichment for mice of the Genus Mus. False

Effect of Environmental Enrichment Devices on Behaviors of Single- and Group-housed Squirrel Monkeys (Saimiri sciureus). Contemporary Topics 36 (3): 72.
Squirrel monkeys display an interest in novel places, habituate to new situations, and spend 90 to 95% of their waking time in the wild in large groups (20 - 200) engaging in feeding behaviors over a broad area (exceeds 250 hectares). The results of the study showed the all monkeys, either group- or single-housed, spent about half of their time being inactive, regardless of whether they had enrichment devices. Both groups spent similar percentage of time (less than 2%) interacting with enrichment devices.
To ensure the effectiveness of any enrichment devices, the balance between being too simple and too difficult and the consequences that interaction with it has on an animal should be established. If the task is too difficult, the animals remain an aroused physiologic state, rather than habituating to the device and returning to the lowered arousal level characterizing a state of inactivity. If the animal habituates the device, the animal may simply respond to the food reward.
The group-housed monkeys spent more time grooming and being active, whereas the single-housed monkeys spent more time exhibiting stereotypical behaviors. The social benefits of group housing should not be overlooked as an effective enrichment.
No questions

Surgical Management of a Nasal Adenocarcinoma in an Olive Baboon (Papio cynanocephalus). Contemporary Topics 36 (3): 76.
Case Report
Clinical: Adult male baboon with respiratory difficulty
Exam: Auscultation - normal lung sounds, inspiratory stridor Rhinoscopy - cranial third of right nostril occluded by a lobulated mass, extensive hemorrhage. Nasal septum displaced laterally into the left nostril
Surgery - longitudinal incision extending below the medial right orbit and through subcutaneous tissue and lateral nasal cartilage. Mass removed.
Baboon recovered, no breathing problems, no metasteses, and doing well 2 years later.
Questions:
(nothing significant in this report, other related questions provided)
1. Name the alpha-herpes virus known to commonly infect baboons. Lesions are most likely to be found where on the baboon?
2. Name the nematode that deposits its embryonic eggs in the nasal cavity or under the cutaneous epithelium of monkeys?
Answers:
1. Herpes Papio 2 (HPV2); genitalia
2. Anatrichosoma cutaneum

Multicystic Lesion of the Jaw in a Rabbit. Contemporary Topics 36 (3): 78.
Solitary mass of the mandible with large central cystic area 2 previous reports of mandibular masses in rabbits: ameloblastoma and mucoepidermoid carcinoma
Histo: multiple stratified squamous to cuboidal epithelial lined cysts [containing serosanguinous fluid] with odontogenic rests and foci of fibroblastic connective tissue resembling periodontal ligament. Concluded that this tumor was a mucoepidermoid carcinoma
No questions

A Simple Technique for Collection of Large Amounts of Blood from Tranquilized or Anesthetized Rabbits. Contemporary Topics 36 (3): 81.
- anesthesia: ketamine hydrochloride, 40 mg/kg of body weight, I.M. and xylazine hydrochloride, 6 mg/kg, I.M.
- sedation: fentanyl citrate (0.05 mg/ml) and droperidol (2.5 mg/ml) =3D Innovar, administered subcutaneously at a dosage of 0.5ml/4 kg.
- clip hair over intermediate auricular artery and swab with alcohol
- 22gauge, doubleheaded needle is inserted 0.5 to 1.0cm into the intermediate auricular artery at 5oangle parallel to the skin, and the evacuated tube is attached to the needle.
- tape cotton ball over phlebotomy site for 10 15 minutes
No questions