EU Directive 2010/63/EU


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Sep 4, 2006
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Cephalopod research and EU Directive 2010/63/EU: Requirements, impacts and ethical review
Jane A. Smith,Paul L.R. Andrews, Penny Hawkins, Susanna Louhimies, Giovanna Ponte, Ludovic Dickel
For the first time, European Union legislation on animal research and testing has extended its scope to include invertebrate species—the Class Cephalopoda. EU Directive 2010/63/EU, which was due to be implemented in Member States 1 January 2013, covers all “live cephalopods” used in scientific procedures that are likely to cause the animals adverse effects such as “pain, suffering, distress or lasting harm”.
This paper examines practical implications of the new EU law for cephalopod research. It evolved from a meeting of European cephalopod researchers held in Naples in 2011 (EuroCeph), which in turn was stimulated by discussions within The Boyd Group (a UK forum on animal experiments). This paper:
1. describes key requirements of Directive 2010/63/EU;
2. explains the project evaluation process that all regulated scientific projects involving animals must undergo before they can be authorised within Member States;
3. presents a series of hypothetical case studies, to illustrate how, in practice, the principles for project evaluation might be applied in cephalopod research and testing;
4. highlights the need for widely agreed guidance specific to cephalopods, to assist regulators, establishments and researchers in implementing the new law; and
5. concludes with a list of practical steps that researchers might take to ensure compliance with the Directive in the national legislation of all EU Member States.
At the airport heading to Kansas City for a conference and just read the paper on a free iPad at Laguardia, which is awesome!!! What isn't awesome is that alcohol is not served until 8am..... To the paper, good review on what the new directive entails and some interesting discussion points as well. However, I'm still at a loss for the reasoning behind the directive.... Were there questionable practices going on????? The reference to the 3 Rs was a good talking point and makes sense. It also makes sense to report how animals were collected from the wild, including methods, mortalities, etc. But I ask, why wasn't this done before? Who wasn't doing this? I was also surprised that the all encompassing directive still does not cover ALL types of experiments and those not covered will be assessed based on a "best judgment" policy. Seems strange given the scope of the directive. The directive definitely brings up some good points but the need for increased regulations and oversight seems a bit much.... Going forward in the USA, I don't believe we need more regulations for cephalopods. Rather, we should utilize the information in the directive for discussion and to simply take another look at our methods and how we conduct experiments. I like the ideas in the directive. I don't like the increased oversight. Greg
Scientists Learn How to Put an Octopus to Sleep
By Katherine Harmon Courage | April 29, 2014 |

We can’t really ask an octopus to count backward from 10. Which is just one of the tricky things about putting an octopus under.

If knocking an octopus out (for science) sounds like an unusual procedure, well, it is. But it’s likely going to get a lot more common in labs around the world.

Canada, the European Union and the U.K. have decided that the octopus, with all of its cognitive and behavioral complexity, should really be treated a bit more carefully in the lab than some of its more cognitively humble mollusk relatives. In fact, the octopus has been declared an honorary vertebrate—as far as its treatment goes.

“Experimentation with animals is always something that has to be handled ethically and appropriately,” Frank Grasso director of Brooklyn College’s Biomemetic and Cognitive Robotics Lab, explained earlier this month at an event in New York City. “And the smarter octopuses appear to us, the more evidence we have for their intelligence, the more we can’t treat them like snails.”

With the increasing insistence that octopuses ought to be granted more consideration, comes the question of just how to make their experience less distressing. Not to mention the urgent scientific question of what effects the anesthesia might have on the octopuses. “This is a new field to work in,” Grasso noted. “And it is an ethical imperative now.”

While we are pretty adept at anesthetizing mice and monkeys, the mollusk body works so differently that human researchers are sill largely in the dark. “We have a tremendous problem in terms of ignorance of knowing what’s right to do with octopuses and wrong,” Grasso said. So his team has been trying to suss out this slippery issue.

As Grasso explained: “The octopuses that have come into my laboratory right now are going through a series of rigorous examinations to be able to assess whether or not these anesthetics—that have been used since the 1930s and have really not been examined—are in fact helpful to the animals, have long-term deleterious effects, and, in fact are ethical to use.”

And ethics come into play simply when handling these animals.

“If an animal is going to be experiencing some kind of procedure that is unpleasant, you would want to minimize the animal’s distress,” Grasso said. Even for un-invasive procedures, an encounter with a scientist is likely no day at the spa for the octopus, which Grasso describes as, basically, “a bowl of jelly.” Simply “the ability to try to hold it steady long enough can be rather difficult. So you like to have a general anesthetic…so that the animal isn’t stressed out by the experimenter wrestling with it.”

In anecdotal observations, Grasso notes, he and his colleagues have noticed behavioral changes in their octopuses at Brooklyn College after an anesthesia treatment. But there is still much to learn about putting octopuses safely to sleep (and waking them back up again).

“This work is so fresh right now, it’s hard to know what’s going to ultimately happen,” Brooklyn College graduate researcher Olivia Mae Davis told The Columbia Chronicle earlier this month. “But it will affect laws, policies, the lives of the animals and humans as well.”
This might be going a bit too far ...

The Use of Artificial Crabs for Testing Predatory Behavior and Health in the Octopus1
Piero Amodio, Paul Andrews, Marinella Salemme, Giovanna Ponte 2014 (pdf)

Summary The willingness of the cephalopod mollusc Octopus vulgaris to attack a live crab is traditionally used as a method to assess the overall health and welfare of octopuses in the laboratory. This method requires placing a crab in the home tank of an animal, measuring the time (latency) taken for the octopus to initiate an attack and withdrawing the crab immediately prior to capture. The same crab is commonly used to assess multiple octopuses as part of daily welfare assessment. Growing concern for the welfare of crustaceans and a review of all laboratory practices for the care and welfare of cephalopods following the inclusion of this taxon in 2010/63/EU prompted a study of the utility of an artificial crab to replace a live crab in the assessment of octopus health. On consecutive days O. vulgaris (N=21) were presented with a live, a dead or an artificial crab, and the latency to attack measured. Despite differences in the predatory performance towards the three different crab alternatives, octopuses readily attacked the artificial (and the dead) crab, showing that they can generalize and respond appropriately towards artificial prey. Researchers should consider using an artificial crab to replace the use of a live crab as part of the routine health assessment of O. vulgaris.
US Animal Welfare Act

EU Directive

Guidelines for the Care and Welfare of Cephalopods in Research –A consensus based on an initiative by CephRes, FELASA and the Boyd Group

Below is a list of ceph sensitivity studies compiled by Eric Edsinger ( @000generic ) in preparation for his doctoral research:

Alupay, J. S., Hadjisolomou, S. P., & Crook, R. J. (2014). Arm injury produces long-term behavioral and neural hypersensitivity in octopus. Neuroscience Letters, 1–6.

Andrews, P. L. R. (2011). Introduction: laboratory invertebrates: only spineless, or spineless and painless? ILAR Journal / National Research Council, Institute of Laboratory Animal Resources, 52(2), 121–125.

Andrews, P. L. R., Darmaillacq, A.-S., Dennison, N., Gleadall, I. G., Hawkins, P., Messenger, J. B., et al. (2013). The identification and management of pain, suffering and distress in cephalopods, including anaesthesia, analgesia and humane killing. Journal of Experimental Marine Biology and Ecology, 447(C), 46–64.

Crook, R. J., & Walters, E. T. (2011a). Nociceptive behavior and physiology of molluscs: animal welfare implications. ILAR Journal / National Research Council, Institute of Laboratory Animal Resources, 52(2), 185–195.

Crook, R. J., Lewis, T., Hanlon, R. T., & Walters, E. T. (2011b). Peripheral injury induces long-term sensitization of defensive responses to visual and tactile stimuli in the squid Loligo pealeii, Lesueur 1821. Journal of Experimental Biology, 214(19), 3173–3185.

Crook, R. J., Hanlon, R. T., & Walters, E. T. (2013). Squid Have Nociceptors That Display Widespread Long-Term Sensitization and Spontaneous Activity after Bodily Injury. Journal of Neuroscience, 33(24), 10021–10026.

Crook, R. J., Dickson, K., Hanlon, R. T., & Walters, E. T. (2014). Nociceptive Sensitization Reduces Predation Risk. Current Biology, 24(10), 1121–1125.

Fiorito, G., Affuso, A., Anderson, D. B., Basil, J., Bonnaud, L., Botta, G., et al. (2014). Cephalopods in neuroscience: regulations, research and the 3Rs. Invertebrate Neuroscience, 14(1), 13–36.

Fiorito, G., Affuso, A., Basil, J., Cole, A., de Girolamo, P., D'Angelo, L., et al. (2015). Guidelines for the Care and Welfare of Cephalopods in Research -A consensus based on an initiative by CephRes, FELASA and the Boyd Group. Laboratory Animals, 49(2 Suppl), 1–90.

Hague, T., Florini, M., & Andrews, P. L. R. (2013). Preliminary in vitro functional evidence for reflex responses to noxious stimuli in the arms of Octopus vulgaris. Journal of Experimental Marine Biology and Ecology, 447(C), 100–105.

Mather, J. A. (2008). Cephalopod consciousness: behavioural evidence. Consciousness and Cognition, 17(1), 37–48.

Moltschaniwskyj, N. A., Hall, K., Lipinski, M. R., Marian, J. E. A. R., Nishiguchi, M., Sakai, M., et al. (2007). Ethical and welfare considerations when using cephalopods as experimental animals. Reviews in Fish Biology and Fisheries, 17(2-3), 455–476.

National Research Council (US) Committee on Recognition and Alleviation of Pain in Laboratory Animals. (2009). Recognition and Alleviation of Pain in Laboratory Animals. Washington (DC): National Academies Press (US).

National Research Council (US) Committee for the Update of the Guide for the Care and Use of Laboratory Animals. (2011). Guide for the Care and Use of Laboratory Animals (8 ed.). Washington (DC): National Academies Press (US).

Polese, G., Winlow, W., & Di Cosmo, A. (2014). Dose-Dependent Effects of the Clinical Anesthetic Isoflurane on Octopus vulgaris: A Contribution to Cephalopod Welfare. Journal of Aquatic Animal Health, 26(4), 285–294.

Smith, J. A., Andrews, P. L. R., Hawkins, P., Louhimies, S., Ponte, G., & Dickel, L. (2013). Cephalopod research and EU Directive 2010/63/EU: Requirements, impacts and ethical review. Journal of Experimental Marine Biology and Ecology, 447(C), 31–45.
Why are cephalopods protected in scientific research in Europe?
Belinda M. Tonkins (full paper on-line)
This essay will discuss the reasons why cephalopods are now protected by UK and EU law when used in scientific procedures, including evidence for their potential sentience and ability to feel pain. It will examine the significant provisions of the law and how it aims to maximise welfare. It will then conclude on the effectiveness of the legislation and potential future directions for invertebrate welfare.
In vivo recording of neural and behavioral correlates of anesthesia induction, reversal, and euthanasia in cephalopod molluscs. Robyn J. Crook, Hanna M Butler-Struben, Samantha M. Brophy, Nasira A. Johnson 2017 (Frontiers in Physiology)

Cephalopod molluscs are among the most behaviorally and neurologically complex invertebrates. As they are now included in research animal welfare regulations in many countries, humane and effective anesthesia is required during invasive procedures. However, currently there is no evidence that agents believed to act as anesthetics produce effects beyond immobility. In this study we demonstrate, for the first time, that two of the most commonly used agents in cephalopod general anesthesia, magnesium chloride and ethanol, are capable of producing strong and reversible blockade of afferent and efferent neural signal; thus they are genuine anesthetics, rather than simply sedating agents that render animals immobile but not insensible. Additionally, we demonstrate that injected magnesium chloride and lidocaine are effective local anesthetic agents. This represents a considerable advance for cephalopod welfare.
Using a reversible, minimally invasive recording procedure, we measured activity in the pallial nerve of cuttlefish (Sepia bandensis) and octopus (Abdopus aculeatus, Octopus bocki), during induction and reversal for five putative general anesthetic and two local anesthetic agents. We describe the temporal relationship between loss of behavioral responses (immobility), loss of efferent neural signal (loss of ‘consciousness’) and loss of afferent neural signal (anesthesia) for general anesthesia, and loss of afferent signal for local anesthesia. Both ethanol and magnesium chloride were effective as bath-applied general anesthetics, causing immobility, complete loss of behavioral responsiveness and complete loss of afferent and efferent neural signal. Cold seawater, diethyl ether and MS-222 (tricaine) were ineffective. Subcutaneous injection of either lidocaine or magnesium chloride blocked behavioral and neural responses to pinch in the injected area, and we conclude that both are effective local anesthetic agents for cephalopods. Lastly, we demonstrate that a standard euthanasia protocol – immersion in isotonic magnesium chloride followed by surgical decerebration – produced no behavioral response and no neural activity during surgical euthanasia. Based on these data, we conclude that both magnesium chloride and ethanol can function as general anesthetic agents, and lidocaine and magnesium chloride can function as local anesthetic agents for cephalopod molluscs.

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