Care and Use of Vertebrate
Animals
Dr. Janet Whaley
Veterinarian for UMCES
IACUC
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My Info
• Aquaculture Program Manager
USDA APHIS Veterinary Services
4700 River Road, Unit 46
Rm. 4B.02.11
Riverdale, MD 20737
Janet.E.Whaley@aphis.usda.gov
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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
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•
•
•
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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 preeuthanasia
• 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-manimobenzoate 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 Diagnostics
Skin scrape
Fin clip
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Simple Diagnostics
Gill clip
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Clinical Diagnostics
Blood collection for analysis
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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,
scombroid, dinoflagellates toxins)
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PATHOGEN
B A C T E R IA
S trep tococcu s
S tap h ylococcu s
C lostrid iu m
E rysip eloth rix
M ycob acteriu m
N ocard ia
V ib rio
P . sh igelloid es
A erom on as
P seu d om on as
E sch erich ia
S alm on ella
K leb siella
E d w ard siella
Lep tosp irosis
P A R A S IT E S
A n asak iasis
E u stron gyloid es
C estod es
T rem atod es
P rotozoa
IN G E S T IO N O F F IS H
T IS S U E (U N D E R C O O K E D
O R F E C E S C O N T .)
IN G E S T IO N O F
IN F E C T E D
A Q U A R IA W A T E R
DERM AL CONTACT
IN F E C T E D
F IS H
*
*
+
+
*
*
+
+
+
*
+
+
+
*
+
*
+
+
+
?
+
?
+
+
+
+
*
*
+
+
?
V IR U S E S
C aliciviru s
*
FUNGI
C an d id a
*
T O X IN S
C igu atera
P oison in g
S com b roid
P oison in g
D E R M A L C O N T A C T IN F E C T E D
A Q U A R IU M /S E A W A T E R
+
?
+
h eat an d cold stab le
+
cold sen sitive
+ rep orted cases in h u m an s
* n o k n ow n cases in h u m an s b u t th e p oten tial risk exists
? exp osu re rou te of rep orted h u m an d isease u n k n ow n
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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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