Amphibians: Guidelines for the breeding, care, and management of laboratory animals

Contents

I Introduction 1
A. General Comments 1
B. The Need 1
1. The Demand, 1
2. The Supply, 3
a. The Natural Resource, 3
b. Artificial Culture, 3
3. Recommendations. 7
C. Users of Amphibians 7
D. Laboratory-Defined Amphibians 11
II Classification and Description of Amphibians Commonly
Used for Laboratory Research 12
A. Phylogeny and Classification 12
B. Description of Species Commonly Used in
Laboratory Research 13
1. Urodeles, 13
a. Ambystoma, 13
b. Notophthalmus wiridescens, 15
c. Necturus maculosus, 16
2. Anurans, 16
a. Xenopuslaevi:, 16
b. Rana catesbeiana, 17
c. Rena grylio, 18
d. Rana clamitans, 18
e. Rana pipiens, 21
f. Rana palustris, 23
g. Rana sylvatica, 24
h. Bufo marinus, 24
i Bombina orientalis, 25

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III Definition and Description of Experimental Amphibians 27
A. Introduction27
B. Definition28
1. Wild, 28
2. Wild Caught, 28
a. Nonconditioned, 28
b. Conditioned, 30
3. Laboratory Reared, 31
a. Standard, 31
b. Miscellaneous, 31
4. Laboratory Bred, 32
a. Standard, 32
b. Miscellaneous, 32
C. Description of Laboratory-Reared and Laboratory-
Bred Amphibians32
1. Types of Populations and Lines, 32
a. Random Mating Lines, 33
b. Heterozygous Isogenic Clones, 33
c. Heterozygous Marked Lines, 33
d. Mutant Lines, 34
e. Inbred Lines, 34
f. Gynogenetic Diploid Lines, 34
g. Homozygous Lines, 34
h. Haploid Animals, 35
i Polyploid Animals, 35
2. Sex Determination and Its Manipulation, 36
3. Species of Laboratory-Reared or Laboratory-Bred
Amphibians Available in the United States, 37
a. Anurans, 37
b. Urodeles, 3
IV Sources 39
A. Selection39
B. Dealer Care40
C. Ordering, Shipping, and Receiving40
1. Ordering,40
2. Shipping,41
3. Receiving, 42
a. Aquatic Amphibians, 42
b. Terrestrial Amphibians, 43
D. Legal Aspects45
V Physical Facilities 46
A. Relation to Natural Habitat46
B. The Amphibian Quarters47
1. General Description, 47
a. Isolation Quarters, 47
b. Heating, Ventilation, and Size Specifications for Rooms, 48

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c. Description of Ancillary Rooms, 49
d. General Specifications, 49
2. Environmental Control, 50
a. Water, 50
b. Temperature, 57
C. Lighting, 57
C. Enclosures 58
1. Embryos: Fertilization through Initiation of Feeding, 58
2. Larvae, 59
3. Juveniles, 60
a. At Metamorphic Climax, 60
b. Postmetamorphic, 61
4. Adult Enclosures (Evaluation Criteria), 62
5. Hibernation Quarters, 63
6. Enclosure Designs, 64
a. The Amphibian Facility of The University of Michigan, 64
b. The R. catesbeiana Facility at Louisiana State University, 68
C. Southern Frog Company, 71
d. The Aquatic Animal Facility of Arizona State University, 72
VI Amphibian Management and Laboratory Care 75
A. General Comments 75
B. Anurans 76
1. Ranidae, 76
a. R. pipiens, 76
b. R. catesbeiana, 84
c. Other Ranid Species, 87
2. Other Anurans, 88
a. Xenopus, 88
b. Bufo, 90
c. Bombina orientalis, 90
C. Urodela 91
1. Axolotis, 91
a. Early Larvae,91
b. Mature Larvae, 92
2. Other Urodeles, 93
VII Breeding 95
A. Anurans 95
1. General Comments, 95
2. Artificial Induction of Ovulation, 95
3. Amplexus, 96
4. Artificial Insemination, 97
5. Parthenogenetic Lines and Haploid Animals, 99
6. Homozygous Lines and Androgenesis, 100
7. Polyploid Animals, 100
8. Mosaic Animals, 101
9. Nuclear Transplantation, 101
10. Xenopus, 101

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B. Urodeles 103
1. Mating, 103
2. Spawning, 104
3. Artificial Insemination, 106
4. Initial Care of Embryos, 107
VIII Records and Information Control 108
A. Identification of Individuals 108
1. Numbeting,108
2. Tattooing, 109
3. Branding, 109
4. ToeClipping,110
5. Other Marking Systems, 110
6. Drawings and Photographs, 111
B. Information Control Systems 111
IX Amphibian Medicine 115
A. General Comments 115
B. Bacterial Diseases 116
1. Pathogens, 116
2. Drug Selection and Administration, 117
3. Identifying Diseased Frogs, 20
4. The Need for Treatment, 121
C. Viral Diseases 121
1. General Comments, 121
2. Lucke' Tumor Herpesvirus (LTHV), 122
3. Amphibian Polyhedral Cytoplasmic Deoxyribovirus (PCDV), 122
4. Lvmphosarcoma Virus of Xenopus, 123
D. Parasitic Diseases 123
E. Mycotic Diseases 124
F. Euthanasia and Anesthesia 124
X Personnel 127
A. General Comments 127
B. Safety Hazards 128
1. Physical, 128
2. Biological, 128
Appendix A Status of Endangered Amphibians 131
Appendix B Control Laws by States and Canadian Provinces 133
References 143

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