INVERTEBRATE  ANESTHETICS AND RELAXANTS: AGENTS AND ORGANISMS

TABLE  1 

This table is an active compendium to which organisms and agents may be added periodically. Organisms are listed alphabetically by genus in the following subdivisions:

Annelida (Polychaeta)
Arthropoda (Crustacea; Cladocera, Stomatopoda, Amphipoda, Decapoda)
Cnidaria (Actinaria)
Echinodermata (Asteroidea, Ophiuroidea, Holothuroidea)
Mollusca (Bivalvia, Cephalopoda, Gastropoda)
Other Aquatic Invertebrates (Entoprocta)

ABBREVIATIONS USED BELOW:

SW = seawater; FASW = filtered artificial seawater
 

OBJ = GENERAL OBJECTIVES:

EX = examination or manipulation, and recovery IM = immobilization for a variety of purposes
IN = inhibition of unplanned spawning, with recovery IS = improving live shipping condition
MI = microscopic observation MP = morphological preservation
RE = removal of living material TR = transportation and recovery

  AGENTS:

AB = 3-aminobenzoic acid ethyl ester (MS222) AM = amyleine HCl
CH = chloral hydrate CF = chloroform
EA = ethanol, ethyl alcohol IA = isobutyl alcohol
KE = ketamine-HCl LI = lidocaine (lignocaine, Xylocaine)
LT = low temperature MC = magnesium chloride
ME = menthol MF = methoxyflurane (Metofane)
MS = magnesium sulfate PE = 2-phenoxyethanol
PP = propylene phenoxetol [see Note on PP identity] PR = procaine (Novocaine)

XY = xylazine-HCl (Rompun)

NOTICE: This information is for professional use only. Handling and use of any chemical agents referred to in this compendium or in the cited literature requires health and safety precautions (safety mask, chemical hood, etc.) as specified on relevant MSDS data sheets or in other documentation. In addition, legal use of some of these agents on certain invertebrate species may be subject to local animal care and use regulations. The editors and contributors, The Biological Bulletin, and the MBL assume no responsibility for any health effects, loss of animals or data, legal issues, or any other problems that may arise due to use of agents in this compendium or in its cited literature.

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Phylum: ANNELIDA

HIGHER TAXON;
SPECIES		 COMMON NAME OBJ AGENT PROCEDURE LIT. & NOTES
 
Class: Polychaeta
Chaetopterus spp. Parchment tube worms EX PP
EA		 late stage larvae: add 1 vol. 0.1% in FASW to 1 vol. SW containing organisms
early stage larvae: 4% in FASW		 Irvine & Martindale, 1999
Pectinaria gouldii Trumpet worm;
ice-cream cone worm		 EX AB
EA		 0.04% in filtered SW
dropwise addition of 50% sol'n to SW containing organisms		 Tweedell, 1996
Polydora cornuta Spionid palp worm EX PP
EA		 late stage larvae: add 1 vol. 0.1% in FASW to 1 vol. SW containing organisms
early stage larvae: 4% in FASW		 Irvine & Martindale, 1999

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   ANESTHETICS and RELAXANTS,

Phylum: ARTHROPODA

HIGHER TAXA;
SPECIES		 COMMON NAME OBJ AGENT PROCEDURE LIT. & NOTES
 
Subphylum: CRUSTACEA
 
Suborder: Cladocera (water fleas)
Daphnia magna Water flea IM MF ~0.1% (v/v) in 100% oxygen bubbled through water McKenzie et al, 1992
 
Order: Stomatopoda (mantis shrimps)
Squilla mantis Mantis shrimp EX IA cardiac injection, 100-200 µl/kg (1 - 2 µl/10 g), 11-17oC, - or -
immersion, 0.04% (v/v) in SW, 17oC, 1 h		 Ferrero and Pressacco, 1982
 
Order: Amphipoda
Gammarus pulex Fresh-water shrimp EX AB 0.1% (w/v), ~20 min at 20 - 23oC; recovery best at 15oC or below Ahmad, 1969
 
Order: Decapoda (shrimp, crayfish, lobsters, crabs)
Carcinus maenas Green crab EX PR 25 mg/kg injection into haemocoel; anes. in 30 s after brief hyperactivity; recovery 2-3 h at 10oC Oswald, 1977
Homarus americanus American lobster EX IA 0.15% - 0.7% (v/v) in aerated filtered SW; agitation precedes anesthesia; complete recovery in SW Foley et al, 1966
Orconectes virilis Crayfish EX LI
KE		 intramuscular injection, >300 µg/g; lasts 20-30 min
intramuscular injection, >90 µg/g; lasts >60 min		 Brown et al, 1996
Penaeus japonicus Kuriama prawn IM LT 12oC Paterson, 1993
Pseudocarcinus gigas Australian giant crab IM XY
KE		 injection, 16 or 22 mg/kg
injection, 0.025-0.1 mg/kg		 Gardner, 1997

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   ANESTHETICS and RELAXANTS,

Phylum: CNIDARIA

HIGHER TAXON;
SPECIES		 COMMON NAME OBJ AGENT PROCEDURE LIT. & NOTES
 
Order: Actinaria (sea anemones)
Adamsia carciniopados
(successful with many other spp; see Note)		   -- MP ME grind crystals to coarse powder and scatter over surface of SW containing organisms Moore, 1989
Calliactis parasitica   -- MI, MP MC 0.3M, until "sufficient" relaxation Westfall et al, 1995

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   ANESTHETICS and RELAXANTS,

Phylum: ECHINODERMATA

HIGHER TAXA;
SPECIES		 COMMON NAME OBJ AGENT PROCEDURE LIT. & NOTES
 
Class: Asteroidea (sea stars, or starfish)
Astropecten articulatus Margined sea star MI, MP MC 7.5% MgCl2.6H20 + SW, 1:1 McCurley and Kier, 1995
Coscinasterias calamaria   -- RE AB 1% in SW, 45 min O'Neill, 1994
Luidia clathrata Grey sea star MI, MP MC as for Astropecten McCurley and Kier, 1995
 
Class: Ophiuroidea (brittle stars)
Amphipholis squamata [formerly Axiognathus squamatus]   -- RE ME 2.5-5% in sterile SW Costello and Henley, 1971
 
Class: Holothuroidea (sea cucumbers)
Holothuria forskali   -- MI, MP PP 2 o/oo sat. sol'n in SW, 1 h VandenSpiegel and Jangoux, 1987
Stichopus badionotus Bermuda sea cucumber MP PP sat. sol'n in SW, 2 h Hill and Reinschmidt, 1976

 

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   ANESTHETICS and RELAXANTS,

Phylum: MOLLUSCA

HIGHER TAXA;
SPECIES		 COMMON NAME OBJ AGENT PROCEDURE LIT. & NOTES
 
Class: Bivalvia (mussels, scallops, oysters, cockles, clams)
Cumingia tellinoides Tellin-like cumingia MI CH a "dilute" sol'n in SW for older larvae Costello and Henley, 1971
Mytilus edulis Edible mussel, blue mussel IM PP sat. sol'n(?) in SW, 30 min [see Note on PP use] Owen, 1955
Ostrea edulis European flat oyster EX MC 3.5% (w/v), ~90 min; recovery ~90 min Culloty and Mulcahy, 1992
Pecten fumatus Commercial scallop (southern Australia) IN CH
MC		 4 g/l (0.024M) for < 20 min
30 g/l (0.31M) for < 10 min		 Heasman et al, 1995
Pinctada maxima Silver-lip pearl oyster EX PP 1.5 ml/l (lab); 2-2.5 ml/l (field) Mills et al, 1997
Spisula subtruncata Thick trough shell (western Europe) IM PP sat. sol'n (?) in SW, 30 min [see Note on PP use] Owen, 1955
 
Class: Cephalopoda (squid, cuttlefish, octopus)
Eledone cirrhosa Horned octopus EX MC 3.75% MgCl2.6H2O in 50% SW; 15oC, 15 min Messenger et al, 1985
Loligo forbesi Veined squid EX MC as E. cirrhosa, except 13oC, 1.5 min Messenger et al, 1985
Loligo pealei Longfin inshore squid (Woods Hole) EX EA 1.5% for brief period O'Dor et al, 1990
Loligo (Alloteuthis) subulata European common squid EX MC as E. cirrhosa, except 14oC, 1.5 min Messenger et al, 1985
Octopus vulgaris Common octopus EX MC as E. cirrhosa, except 22oC, 13 min Messenger et al, 1985
Sepia officinalis Common cuttlefish EX MC as E. cirrhosa, except 15oC, 5-12 min Messenger et al, 1985
Sepioteuthis sepioidea Caribbean reef squid EX EA
MC
MS		 1%-3% in filtered SW, aerated, "room temp."
as above, except 1.5%-2%
as above, except 3%-4%		 Garcia-Franco, 1992
Todarodes pacificus Japanese flying (or common) squid TR LT 0-1oC SW, "several" min - 6.5 h Bower et al, 1999
 
Class: Gastropoda (snails, slugs, whelks, conchs, abalone, etc.)
Cerastoderma (= Cardium) edule Edible cockle IM PP sat. sol'n(?) in SW, 30 min [see Note on PP use] Owen, 1955
Crepidula fornicata Slipper limpet MI CH a "dilute" sol'n for swimming larvae Costello and Henley, 1971
Haliotis midae Midas ear abalone (South Africa) EX MS
PE		 4%-22% (w/v), 5-8 min
0.05%-0.3% (v/v), 1-2 min		 White et al, 1996

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   ANESTHETICS and RELAXANTS,

OTHER AQUATIC INVERTEBRATES

HIGHER TAXON;
SPECIES		 COMMON NAME OBJ AGENT PROCEDURE LIT. & NOTES
 
Phylum: ENTOPROCTA
Loxosomella brochobola Antarctic entoproct MI, MP AM -> MC "several grains" AM in ~3 ml SW with organisms;
when fully expanded, replace with isotonic MC		 Emscherman, 1993