Care and Use of Vertebrate Animals
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Care and Use of Vertebrate Animals
Dr. Janet WhaleyVeterinarian for UMCES
IACUC
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My Info
• Aquaculture Program ManagerUSDA APHIS Veterinary Services4700 River Road, Unit 46
Rm. 4B.02.11Riverdale, MD [email protected]
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Vet Duties• UMCES Assurance of Compliance with
the Public Health Service Policy on Humane Care and Use of Laboratory Animals -
• advise on appropriate procedures for use of finfish in research, review research proposals, inspect UMCES facilities, and provide annual training.
• authority to suspend any research found to be in violation of UMCES or PHS policy.
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Justification
• Research Goals• Non-animal alternatives• Duplication • Research species• Animal use and pain category
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Pain and Fish
• sensory receptors are present• central reception of sensory input is
unclear• clinical signs of acute and chronic stress
can be observed (cortisol levels, changes in other health and behavior parameters)
• avoid adverse stimuli (reflex manner)
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Pain Category
• Category 1 little or momentary pain (euthanasia, tagging)
• Category 2 potential pain or discomfort relieved by anesthetic (euthanasia, surgical procedure)
• Category 3 discomfort or pain which is not relieved
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STRESS
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Causes of Stress
• water quality (O2, ammonia, nitrite, pH, other contaminants)
• transportation• netting &
handling • temperature• salinity• water hardness
• poor nutrition• inappropriate
housing conditions• noise• lighting• vibrations• stocking density
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Stress Response
• Sympathetic nervous system activation
• cortisol• catecholamines• HR, RR• serum osmolality• glucose
• Immunosuppression
( disease resistance)• growth rate• reproduction rate• delayed “capture”
mortality
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Experimental Procedures
• Experimental design (include statistical methodology for data analysis and determination of number of animals to be used)
• Methods and Materials (describe specifically any handling procedures)
• Methods for anesthesia and euthanasia
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Anesthesia• Consider for painful/stressful procedures and pre-
euthanasia
• Ice water (transport) - be careful
• Chemical - MS-222, Benzocaine
• Dose is species specific
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Immersion Anesthesia
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MS-222• tricaine methanesulfate, ethyl-m-
animobenzoate methansulfate, Finquel®• CNS depressant• water soluble but acidic (add buffer)• admin. via bath or recirculating system• for anesthesia 50-100 mg/L recommended
(sedation vs surgical)• dose may be species specific - test before
experiment
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MS-222 con’t
• induction w/in 3 minutes• recovery w/in 10-15 minutes after removal • is residual +/- could affect chemical analysis of
tissue• no known hazards but wear gloves! • list as chemical hazard in UMCES application
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Levels of Anesthesia
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Euthanasia
• +/- pre-sedation with MS-222• decapitation• pithing• chemical (MS-222)• requires experience!!!• avoid direct insertion into fixative (alcohol or
formalin)
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Husbandry Practices
• Briefly describe housing, feeding, etc. (refer to specific laboratory standard operating procedures)
• Disposition of alive and dead animals
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Water Quality
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The Importance of Good Water Quality
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• Investigate mortality in your system
• Basic necropsy• Know your species• Seek advice• Make appropriate
changes
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Simple DiagnosticsSkin scrape
Fin clip 21
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Simple Diagnostics
Gill clip 22
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Clinical Diagnostics
Blood collection for analysis 23
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Environmental Safety
• infectious agents• chemical hazards (include MS-222)• radioisotopes• biohazards
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ZOONOSIS• Zoonosis = disease that can be transmitted
from animals to humans (or other animals)• Anthroponosis = disease that can be
transmitted from humans to animals• High Risk
– immunosupressed (AIDs, other debilitating disease)– pregnant– age
• Exposure (infected water, fish tissue, fish excrement)– dermal contact via skin abrasion, fissure– ingestion
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Zoonotic Diseases • Potential for disease organisms to spread between
species (fish human)• Bacteria - from handling (mycobacterium,
streptococcus, erysipelothrix, vibrio, norcardia, aeromonas, edwardsiella)
from ingestion (stahylcoccus, clostridium, vibrio, aeromonas, esherichia, salmonella,
edwardsiella)• Parasites - primarily from ingestion (nematodes,
cestodes, trematodes, protozoa).• Toxins - primarily from ingestion (ciguatera,
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PATHOGEN INGESTION OF FISH TISSUE (UNDER COOKED OR FECES CONT.)
INGESTION OF INFECTED AQUARIA WATER
DERMAL CONTACT INFECTED FISH
DERMAL CONTACT INFECTED AQUARIUM/SEA WATER
BACTERIA Streptococcus * Staphylococcus * Clostridium + Erysipelothrix + Mycobacterium * + Nocardia * * Vibrio + + P. shigelloides + +
Aeromonas + + Pseudomonas * * Escherichia + Salmonella + Klebsiella + Edwardsiella + + + + Leptospirosis ? ? ? ? PARASITES Anasakiasis + Eustrongyloides + Cestodes + Trematodes + Protozoa * * VIRUSES Calicivirus * FUNGI Candida * TOXINS Ciguatera Poisoning
+ heat and cold stable
Scombroid Poisoning
+ cold sensitive
+ reported cases in humans * no known cases in humans but the potential risk exists ? exposure route of reported human disease unknown
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Mycobacteria
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PREVENTION• Fish
• Know health of your fish• Proper husbandry/aquaculture• Minimum - wear gloves when handling
• All Wildlife• Know the hazards • Take all necessary “known”precautions• DO YOUR HOMEWORK!!!
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References• Can Fish Suffer?: perspectives on sentience,
pain, fear and stress; K.P. Chandroo et al./Applied Animal Behaviour Science 86 (2004) 225-250
• Fish Cognition and Behavior; Culum Brown et al.; Blackwell Publishing (2006) ISBN: 9781405134293
• Fish Medicine; Michael Stoskopf, W.B. Saunders Company; 1st edition (January 15, 1993) ISBN: 0721626297
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