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