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. 2019 Jan 1;58(1):58-64.
doi: 10.30802/AALAS-JAALAS-18-000040. Epub 2018 Nov 29.

Anesthesia and Euthanasia of Brine Shrimp (Artemia franciscana)

Amanda K Darbyshire  1 , Kendra H Oliver  2 , William D Dupont  3 , W Dale Plummer  3 , Carissa P Jones  4 , Kelli L Boyd  5
Affiliations

Anesthesia and Euthanasia of Brine Shrimp (Artemia franciscana)

Amanda K Darbyshire et al. J Am Assoc Lab Anim Sci. .

Abstract

Invertebrates are often overlooked as laboratory animals, yet they are commonly used in toxicology, developmental, cellular and molecular biology, and radiation studies with euthanasia as an endpoint. Little is known regarding appropriate euthanasia methods for invertebrate species, particularly for Artemia. Here, we evaluated the AVMA-recommended 2-step method of euthanasia in brine shrimp (Artemia franciscana). Artemia were exposed first to anesthetic solutions of 60% alcohol, 2.5 mg/L eugenol, or 4 g/L tricaine methanesulfonate (TMS) and then were transferred to euthanasia solutions of 70% alcohol, 95% alcohol, or 10% neutral buffered formalin. We examined time to anesthesia, behavioral response to anesthesia, anesthesia recovery, and time to euthanasia. Our results show that 2.5 mg/L eugenol and 4 g/L TMS inconsistently achieved anesthesia. Although 60% alcohol produced anesthesia, the time to anesthesia varied among replicate groups, and exposure resulted in an increase in abnormal behavior. We therefore do not recommend any of the tested anesthetic solutions for use in Artemia. Although all 3 euthanasia solutions were effective, more research is needed to provide recommendations regarding euthanasia for this species.

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Figures

Figure 1.
Figure 1.
Artemia were placed in 1 mL of anesthetic solution in a 24-well plate according to a permuted plate randomization design.
Figure 2.
Figure 2.
(A). TMS achieved anesthesia in 63% of Artemia; 60% alcohol achieved anesthesia in nearly 100%. Time to anesthesia differed significantly (P = 0.0000) among all groups. (B). Survival curves for euthanasia after anesthesia show 100% probability of death for all groups. All 3 solutions achieved euthanasia, but 95% alcohol led to the fastest euthanasia. (C). Total euthanasia time from placement into anesthetic, 5 min of anesthesia time, until euthanasia in the second-step solution. Euthanasia was fastest by using 60% alcohol followed by 95% alcohol.
Figure 3.
Figure 3.
The incidence of abnormal behavior scores was significantly (*, P = 0.023) greater in the alcohol group compared with the control group. These behaviors were significantly (+, P = 0.0005) reduced in the eugenol group compared with the control group.
See this image and copyright information in PMC

References

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