Anaesthetics and Analgesics Used in Aquatic Animals

A special issue of Biology (ISSN 2079-7737). This special issue belongs to the section "Zoology".

Deadline for manuscript submissions: closed (30 November 2022) | Viewed by 30052

Special Issue Editors

i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal
Interests: anaesthesia; behaviour; refinement; animal welfare; fish, zebrafish
Department of Biological & Environmental Sciences, University of Gothenburg, Box 463, SE-405 30 Gothenburg, Sweden
Interests: animal behavior; animal welfare; analgesia; pain; fear; stress; fish; zebrafish; rainbow trout; physiology; neurobiology
Department of Biology, San Francisco State University, 1600 Holloway Avenue, San Francisco, CA 94132, USA
Interests: pain; evolution; neuroscience; nociception; sensitization; welfare; anesthesia; analgesia; refinement; cephalopods; invertebrates

Special Issue Information

Dear Colleagues, 

Fish and other aquatic animals are used for production in aquaculture and the ornamental industry but are also important models in research. The species used are quite diverse in physiology, size, housing conditions, etc., but all have the ability to experience pain in common, as widely demonstrated in the literature. Thus, whenever a fish needs to be weighed, marked, graded, transported, vaccinated, or have any other procedure involving handling or invasive procedures such as surgery, they may be stressed and/or experience pain, compromising fish health, welfare, and productivity, as well as scientific data. Fish are not the only aquatic animals which have been described to experience pain; hence, cephalopods and crustaceans should also be considered. Thus, the use of anesthesia and/or analgesia is crucial. There are several papers regarding the use of these drugs in fishes; however, because of the diversity of fishes and procedure type, more research is needed to refine the most scientifically appropriate choice of anesthetic and/or analgesic protocol for each situation. Much less research is available regarding crustaceans and cephalopods species. Additionally, the potential side effects of these drugs on physiology, behavior, and other molecular alterations must be considered since this could confound data collection.

The purpose of this Special Issue is to provide an interdisciplinary approach not only to understand the efficacy of anesthesia and analgesia but also to study if the drugs used may compromise aquatic animals’ behavior, physiology, and overall health. The use of anesthesia and analgesia represents an important refinement and should be used so that fish, crustacean, and cephalopod welfare can actually be improved during experiments. Moreover, analgesia is poorly studied in these aquatic animals, with a lack of knowledge regarding its real efficacy and pharmacokinetics in different species. This Special Issue invites original research papers and reviews within the scope of gathering scientific research to propose effective and scientifically informed anesthetic and/or analgesic protocols to be used in different situations and different fish, crustacean, and cephalopod species. These studies can help us to inform and develop appropriate sedation and pain relief protocols that can be used by scientists and veterinarians.

Dr. Ana Maria Valentim
Dr. Lynne Sneddon
Dr. Robyn J. Crook
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Biology is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • anesthesia
  • analgesia
  • refinement
  • welfare
  • research
  • aquaculture
  • clinical parameters
  • biochemical assays
  • physiology
  • behavior
  • pharmacokinetics
  • fish
  • crustacean
  • cephalopods

Published Papers (10 papers)

Order results
Result details
Select all
Export citation of selected articles as:

Research

Jump to: Review, Other

18 pages, 2994 KiB  
Article
Evaluation of Candidates for Systemic Analgesia and General Anesthesia in the Emerging Model Cephalopod, Euprymna berryi
by Skyler Deutsch, Rachel Parsons, Jonathan Shia, Sarah Detmering, Christopher Seng, Alyssa Ng, Jacqueline Uribe, Megan Manahan, Amanda Friedman, Gabrielle Winters-Bostwick and Robyn J. Crook
Biology 2023, 12(2), 201; https://0-doi-org.brum.beds.ac.uk/10.3390/biology12020201 - 28 Jan 2023
Cited by 2 | Viewed by 1935
Abstract
Cephalopods’ remarkable behavior and complex neurobiology make them valuable comparative model organisms, but studies aimed at enhancing welfare of captive cephalopods remain uncommon. Increasing regulation of cephalopods in research laboratories has resulted in growing interest in welfare-oriented refinements, including analgesia and anesthesia. Although [...] Read more.
Cephalopods’ remarkable behavior and complex neurobiology make them valuable comparative model organisms, but studies aimed at enhancing welfare of captive cephalopods remain uncommon. Increasing regulation of cephalopods in research laboratories has resulted in growing interest in welfare-oriented refinements, including analgesia and anesthesia. Although general and local anesthesia in cephalopods have received limited prior study, there have been no studies of systemic analgesics in cephalopods to date. Here we show that analgesics from several different drug classes may be effective in E. berryi. Buprenorphine, ketorolac and dexmedetomidine, at doses similar to those used in fish, showed promising effects on baseline nociceptive thresholds, excitability of peripheral sensory nerves, and on behavioral responses to transient noxious stimulation. We found no evidence of positive effects of acetaminophen or ketamine administered at doses that are effective in vertebrates. Bioinformatic analyses suggested conserved candidate receptors for dexmedetomidine and ketorolac, but not buprenorphine. We also show that rapid general immersion anesthesia using a mix of MgCl2 and ethanol was successful in E. berryi at multiple age classes, similar to findings in other cephalopods. These data indicate that systemic analgesia and general anesthesia in Euprymna berryi are achievable welfare enhancing interventions, but further study and refinement is warranted. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)
Show Figures

Figure 1

10 pages, 37091 KiB  
Article
Electroencephalographic Response in Juvenile Tambaqui, Colossoma macropomum, Exposed to Short-Term Anaesthetic Baths with Geraniol and Citronellol
by Ednara Ronise Lima De Araújo, Marcelo Ferreira Torres, Brenda Maria Pereira Alho Da Costa, Moisés Hamoy, Luís André Sampaio and Luis André Luz Barbas
Biology 2023, 12(1), 90; https://0-doi-org.brum.beds.ac.uk/10.3390/biology12010090 - 06 Jan 2023
Cited by 3 | Viewed by 1296
Abstract
The aim of this study was to evaluate the level of neuronal depression in juvenile tambaqui, Colossoma macropomum, exposed to geraniol (GRL) and citronellol (CTL) in immersion baths. A total of 36 juveniles weighing 35.2 ± 9.4 g were used, organised into [...] Read more.
The aim of this study was to evaluate the level of neuronal depression in juvenile tambaqui, Colossoma macropomum, exposed to geraniol (GRL) and citronellol (CTL) in immersion baths. A total of 36 juveniles weighing 35.2 ± 9.4 g were used, organised into six experimental groups: I—control (clean water); II—ethanol (water containing the highest volume of ethanol used in the anaesthetic pre-dilution); III—GRL induction (70 µL·L−1); IV—CTL induction (90 µL·L−1); V—GRL recovery; VI—CTL recovery. Electroencephalographic (EEG) recordings were performed for 300 s in each group. EEG tracings of the control and ethanol groups showed regular and similar activity. Upon exposure to the anaesthetics, irregularities were observed in the tracings showing neuronal excitability and increased amplitudes, mainly in the case of CTL. Overall, GRL-exposed fish showed depression of the central nervous system with low and regular tracings throughout induction, presenting a gradual recovery and stable tracings, which were consistent with an adequate general anaesthetic effect. On the other hand, fish exposed to CTL showed altered EEG activity during induction, that could be considered incompatible with an appropriate anaesthetic effect and smooth recovery, presenting high and irregular EEG tracing amplitudes. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)
Show Figures

Figure 1

15 pages, 1522 KiB  
Article
Behavioural Aversion and Cortisol Level Assessment When Adult Zebrafish Are Exposed to Different Anaesthetics
by Jorge M. Ferreira, Sara Jorge, Luís Félix, Gabriela M. Morello, I. Anna S. Olsson and Ana M. Valentim
Biology 2022, 11(10), 1433; https://0-doi-org.brum.beds.ac.uk/10.3390/biology11101433 - 30 Sep 2022
Cited by 2 | Viewed by 2073
Abstract
The use of zebrafish (Danio rerio) as an animal model is growing and occurs in a wide range of scientific areas. Therefore, researchers need better and more appropriate anaesthetics for stressful and/or painful procedures to prevent unpleasant experiences. Thus, we aimed [...] Read more.
The use of zebrafish (Danio rerio) as an animal model is growing and occurs in a wide range of scientific areas. Therefore, researchers need better and more appropriate anaesthetics for stressful and/or painful procedures to prevent unpleasant experiences. Thus, we aimed to study if adult zebrafish displayed aversion-associated behaviours (conditioned place aversion) and alterations in cortisol levels when exposed to equipotent concentrations of MS222, propofol/lidocaine, clove oil, or etomidate. Adult AB zebrafish (mixed-sex, N = 177) were randomly assigned to MS222 (150 mg/L), Propofol/Lidocaine (5 mg/L propofol + 150 mg/L lidocaine), Clove Oil (45 mg/L), or Etomidate (2 mg/L) groups. The conditioned place aversion test was used to assess behavioural aversion. Only etomidate resulted in a similar aversion to the positive control group (HCl; pH = 3). Cortisol levels were measured 5 and 15 min after loss of equilibrium. Etomidate induced low levels of cortisol by impairing its synthesis, whereas all the other groups had similar cortisol levels. Based on our data, etomidate was ruled out as an alternative to MS222, as it showed an aversive profile. The remaining protocols were not innocuous, displaying a weak aversive profile when compared to the positive control. In conclusion, a combination of propofol with lidocaine, clove oil, and MS222 were valid candidates for use as anaesthetic protocols. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)
Show Figures

Figure 1

20 pages, 2538 KiB  
Article
A FELASA Working Group Survey on Fish Species Used for Research, Methods of Euthanasia, Health Monitoring, and Biosecurity in Europe, North America, and Oceania
by Jean-Philippe Mocho and Kristine von Krogh
Biology 2022, 11(9), 1259; https://0-doi-org.brum.beds.ac.uk/10.3390/biology11091259 - 24 Aug 2022
Cited by 5 | Viewed by 3776
Abstract
An international survey was conducted regarding species used for research, methods of euthanasia, health monitoring, and biosecurity in fish laboratories. A total of 145 facilities from 23 countries contributed. Collectively, over 80 different species (or groups of species) were reported to be used [...] Read more.
An international survey was conducted regarding species used for research, methods of euthanasia, health monitoring, and biosecurity in fish laboratories. A total of 145 facilities from 23 countries contributed. Collectively, over 80 different species (or groups of species) were reported to be used for research, of which zebrafish (Danio rerio) was the most common by far. About half of the participating laboratories used multiple species. Anesthetic overdose was the preferred method for euthanasia for adult, fry (capable of independent feeding), and larval (not capable of independent feeding) fish. For all developmental stages, the most popular anesthetic compound was tricaine (MS-222), a substance associated with distress and aversion in several species. Moreover, around half of the respondents did not perform a completion method to ensure death. One-quarter of the responding facilities did not have a health monitoring system in place. While most respondents had some form of quarantine process for imported fish, only a small fraction reported quarantine routines that ensure reliable biological barriers. Furthermore, less than one in five screened fish for pathogens while in quarantine. In sum, there was little consensus amongst facilities in how to perform biosecurity measures. Regarding euthanasia, health monitoring, and biosecurity processes, there is a need for updated and universal guidelines and for many laboratories to adjust their practices. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)
Show Figures

Figure 1

17 pages, 1984 KiB  
Article
A Multi-Site Assessment of Anesthetic Overdose, Hypothermic Shock, and Electrical Stunning as Methods of Euthanasia for Zebrafish (Danio rerio) Embryos and Larvae
by Jean-Philippe Mocho, Florian Lang, Guillaume Valentin, Sébastien Bedu, Robin McKimm, Juan Ramos, Yolanda Saavedra Torres, Sarah E. Wheatley, Joseph Higgins, Mollie E. Millington, Pia Rengtved Lundegaard, Rubén Chamorro Valverde, Vlasta Jenčič and Kristine von Krogh
Biology 2022, 11(4), 546; https://0-doi-org.brum.beds.ac.uk/10.3390/biology11040546 - 01 Apr 2022
Cited by 3 | Viewed by 4107
Abstract
Euthanasia in zebrafish (Danio rerio) younger than 5 days post fertilization (dpf) is poorly described in the literature, and standardized protocols are lacking, most likely because larvae not capable of independent feeding are often not protected under national legislations. We assessed [...] Read more.
Euthanasia in zebrafish (Danio rerio) younger than 5 days post fertilization (dpf) is poorly described in the literature, and standardized protocols are lacking, most likely because larvae not capable of independent feeding are often not protected under national legislations. We assessed the euthanasia efficacy in laboratories in different countries of a one hour anesthetic overdose immersion with buffered lidocaine hydrochloride (1 g/L, with or without 50 mL/L of ethanol), buffered tricaine (1 g/L), clove oil (0.1%), benzocaine (1 g/L), or 2-phenoxyethanol (3 mL/L), as well as the efficacy of hypothermic shock (one hour immersion) and electrical stunning (for one minute), on zebrafish at <12 h post fertilization (hpf), 24 hpf, and 4 dpf. Based on the survival/recovery rates 24 h after treatment, the most effective methods were clove oil, lidocaine with ethanol, and electrical stunning. For 4 dpf larvae, signs of aversion during treatment demonstrated that all anesthetics, except lidocaine, induced aversive behavior. Therefore, the most suited euthanasic treatment was lidocaine hydrochloride 1 g/L, buffered with 2 g/L of sodium bicarbonate and mixed with 50 mL/L of ethanol, which euthanized both embryos and larvae in an efficient and stress-free manner. Electrical stunning also euthanized embryos and larvae efficiently and without signs of aversion; this method needs further assessment in other laboratories to draw firm conclusions. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)
Show Figures

Graphical abstract

21 pages, 1388 KiB  
Article
MS-222 and Propofol Sedation during and after the Simulated Transport of Nile tilapia (Oreochromis niloticus)
by Luís Félix, Rita Correia, Rita Sequeira, Cristiana Ribeiro, Sandra Monteiro, Luís Antunes, José Silva, Carlos Venâncio and Ana Valentim
Biology 2021, 10(12), 1309; https://0-doi-org.brum.beds.ac.uk/10.3390/biology10121309 - 10 Dec 2021
Cited by 5 | Viewed by 3021
Abstract
The use of anesthetics has been suggested as a strategy to hamper live fish transport-induced stress. Still, there is insufficient data available on the use of alternative anesthetics to MS-222. This study investigated the use of propofol to mitigate stress in Nile tilapia [...] Read more.
The use of anesthetics has been suggested as a strategy to hamper live fish transport-induced stress. Still, there is insufficient data available on the use of alternative anesthetics to MS-222. This study investigated the use of propofol to mitigate stress in Nile tilapia (Oreochromis niloticus, 143.8 ± 20.9 g and 20.4 ± 0.9 cm) during a 6 h simulated transport. Individuals (n = 7) were divided into three groups: control, 40 mg L−1 MS-222, and 0.8 mg L−1 propofol. A naïve group non-transported was also considered. During the 6 h transport and 24 h after, the response to external stimuli, opercular movements, water quality parameters, behavior, blood hematology and other physiological values, the histopathology of the gills, the quality of the fillet, and oxidative-stress changes in gills, muscle, brain, and liver were evaluated. Propofol increased swimming activity of fish but decreased opercular movements and responses to external stimuli, indicating oscillations of the sedation depth. Water pH and glucose levels increased, while hematocrit (HCT) and lactate decreased in propofol groups at 6 h. At this time-point, MS-222 also induced a decrease in the HCT and lactate levels while increasing cortisol levels. Despite these effects, the stress-related behaviors lessened with anesthetics compared to the control group. After the recovery period, physiological responses normalized in animals from both anesthetic groups, but the control still had high cortisol levels. Overall, propofol is a good alternative for the transportation of this species, showing efficient sedation without compromising health or fillet quality. However, further pharmacodynamics and pharmacokinetics knowledge is required to support its use in aquaculture settings. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)
Show Figures

Figure 1

19 pages, 3092 KiB  
Article
Screening of Anaesthetics in Adult Zebrafish (Danio rerio) for the Induction of Euthanasia by Overdose
by Kristine von Krogh, Joseph Higgins, Yolanda Saavedra Torres and Jean-Philippe Mocho
Biology 2021, 10(11), 1133; https://0-doi-org.brum.beds.ac.uk/10.3390/biology10111133 - 04 Nov 2021
Cited by 8 | Viewed by 3745
Abstract
Zebrafish are often euthanized by overdose of anaesthesia. However, fish may have aversion towards some anaesthetics, and protocol efficacy varies between species. Using wild type adult Danio rerio, we assessed time to loss of opercular beat, righting, and startle reflexes during induction [...] Read more.
Zebrafish are often euthanized by overdose of anaesthesia. However, fish may have aversion towards some anaesthetics, and protocol efficacy varies between species. Using wild type adult Danio rerio, we assessed time to loss of opercular beat, righting, and startle reflexes during induction of anaesthetic overdose by either tricaine (0.5 g/L or 1 g/L), benzocaine (1 g/L), 2-phenoxyethanol (3 mL/L), clove oil (0.1%), isoeugenol (540 mg/L), lidocaine hydrochloride (1 g/L), or etomidate (50 mg/L). Initial screening demonstrated that benzocaine and buffered lidocaine hydrochloride achieved the fastest loss of reflexes. The rapid induction times were confirmed when retesting using larger batches of fish. The fastest induction was obtained with 1 g/L lidocaine hydrochloride buffered with 2 g/L NaHCO3, in which all adult zebrafish lost reflexes in less than 2 min. Next, we monitored signs of distress during benzocaine or buffered lidocaine hydrochloride overdose induction. The results indicated that buffered lidocaine hydrochloride caused significantly less aversive behaviors than benzocaine. Finally, we tested several buffers to refine the lidocaine hydrochloride immersion. The most efficient buffer for euthanasia induction using 1g/L lidocaine hydrochloride was 2 g/L NaHCO3 with 50 mL/L 96% ethanol, inducing immobility in less than 10 s and with only 2% of adult zebrafish displaying aversive behaviors during treatment. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)
Show Figures

Figure 1

Review

Jump to: Research, Other

42 pages, 693 KiB  
Review
Methods to Induce Analgesia and Anesthesia in Crustaceans: A Supportive Decision Tool
by Guiomar Rotllant, Pol Llonch, José A. García del Arco, Òscar Chic, Paul Flecknell and Lynne U. Sneddon
Biology 2023, 12(3), 387; https://0-doi-org.brum.beds.ac.uk/10.3390/biology12030387 - 28 Feb 2023
Cited by 5 | Viewed by 2623
Abstract
Methods to induce analgesia and anesthesia for research purposes, handling, transport, or stunning have been used in 71 species of crustaceans. A non-systematic literature search was conducted on crustacean anesthetic methods. This review presents a comprehensive evaluation of drugs and non-chemical methods used [...] Read more.
Methods to induce analgesia and anesthesia for research purposes, handling, transport, or stunning have been used in 71 species of crustaceans. A non-systematic literature search was conducted on crustacean anesthetic methods. This review presents a comprehensive evaluation of drugs and non-chemical methods used to provide analgesia and anesthesia in many crustacean species rather than just decapod crustaceans. This information allows users to select an appropriate method or agent for their species of interest. We prepared an on-line tool based on datasette, a no-code open-source solution for simple web-based database frontends that allows exploration and downloading data by method, analgesic/anesthetic, species, life stage, or sex, as well as other data including environmental conditions (temperature, salinity, light), route of administration, dosage, and induction and recovery times. These values can be selected to filter the dataset and export it to CSV or JSON formats. Currently, several techniques and chemicals are, in our opinion, unsuitable for use as anesthetics in crustaceans, and the basis for these opinions are presented. Given the evidence of a pain-like experience in crustaceans, we propose that researchers should treat crustaceans humanely, applying the principles of good handling, care, and the management of stress and pain to safeguard their welfare. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)

Other

Jump to: Research, Review

9 pages, 951 KiB  
Brief Report
Impact of Lidocaine on Pain-Related Grooming in Cuttlefish
by Tzu-Hsin Kuo, Lynne U. Sneddon, Joseph W. Spencer and Chuan-Chin Chiao
Biology 2022, 11(11), 1560; https://0-doi-org.brum.beds.ac.uk/10.3390/biology11111560 - 24 Oct 2022
Cited by 2 | Viewed by 1235
Abstract
Nociception is the neural process of encoding noxious stimuli and is typically accompanied by a reflex withdrawal response away from the potentially injurious stimulus. Studies on nociception in cephalopods have so far focused on octopus and squid, with no investigations to our knowledge [...] Read more.
Nociception is the neural process of encoding noxious stimuli and is typically accompanied by a reflex withdrawal response away from the potentially injurious stimulus. Studies on nociception in cephalopods have so far focused on octopus and squid, with no investigations to our knowledge on cuttlefish. Yet, these are an important species both in scientific and commercial use. Therefore, the present study demonstrated that a standard pain stimulus, acetic acid, induced grooming behaviour directed towards the injection site in cuttlefish and that the injection of lidocaine reduces grooming behaviours in acetic-acid-injected cuttlefish. Wound-directed behaviour demonstrates that the animal is aware of the damage; thus, when subjecting these animals to any painful treatments in the laboratory, researchers should consider alleviating pain by the administration of pain-relieving drugs. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)
Show Figures

Figure 1

10 pages, 671 KiB  
Guidelines
Good Anesthesia Practice for Fish and Other Aquatics
by Aurora Brønstad
Biology 2022, 11(9), 1355; https://0-doi-org.brum.beds.ac.uk/10.3390/biology11091355 - 15 Sep 2022
Cited by 3 | Viewed by 3792
Abstract
Fish and other aquatic animals represent a significant number of species with diverse physiology, size, and housing condition needs. Anesthesia may be necessary for several husbandry procedures as well as treatment of diseases, surgery, or experimental procedures. Choice of drugs and detailed procedures [...] Read more.
Fish and other aquatic animals represent a significant number of species with diverse physiology, size, and housing condition needs. Anesthesia may be necessary for several husbandry procedures as well as treatment of diseases, surgery, or experimental procedures. Choice of drugs and detailed procedures for anesthesia must be adapted to the species in question—there is no “one size fits all” solution. However, there are some basic principles that apply for good anesthetic practice of all animals. These principles include the preparations of animals, personnel, facilities and equipment, monitoring animals under anesthesia, as well as post-anesthetic care to be sure that animals are not lost in the recovery phase. Good anesthesia practice also includes the competence and commitment of personnel involved. Based on professional judgement, key factors will be the focus of this text. Full article
(This article belongs to the Special Issue Anaesthetics and Analgesics Used in Aquatic Animals)
Show Figures

Graphical abstract

Back to TopTop