Transcript MS-222
Care and Use of Vertebrate Animals Dr. Janet Whaley Veterinarian for UMCES IACUC 1 My Info • Aquaculture Program Manager USDA APHIS Veterinary Services 4700 River Road, Unit 46 Rm. 4B.02.11 Riverdale, MD 20737 [email protected] 2 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. 3 Justification • • • • • Research Goals Non-animal alternatives Duplication Research species Animal use and pain category 4 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) 5 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 6 STRESS 7 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 8 Stress Response • Sympathetic nervous system activation • cortisol • catecholamines • HR, RR • serum osmolality • glucose • Immunosuppression ( disease resistance) • growth rate • reproduction rate • delayed “capture” mortality 9 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 10 Anesthesia • Consider for painful/stressful procedures and preeuthanasia • Ice water (transport) - be careful • Chemical - MS-222, Benzocaine • Dose is species specific 11 Immersion Anesthesia 12 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 13 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 14 Levels of Anesthesia 15 Euthanasia • • • • • • +/- pre-sedation with MS-222 decapitation pithing chemical (MS-222) requires experience!!! avoid direct insertion into fixative (alcohol or formalin) 16 Husbandry Practices • Briefly describe housing, feeding, etc. (refer to specific laboratory standard operating procedures) • Disposition of alive and dead animals 17 Water Quality 18 The Importance of Good Water Quality 19 • Investigate mortality in your system • Basic necropsy • Know your species • Seek advice • Make appropriate changes 20 Simple Diagnostics Skin scrape Fin clip 21 Simple Diagnostics Gill clip 22 Clinical Diagnostics Blood collection for analysis 23 Environmental Safety • • • • infectious agents chemical hazards (include MS-222) radioisotopes biohazards 24 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 25 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) 26 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 27 Mycobacteria 28 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!!! 29 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 30