GPO: Tank size.

[Thales]

I hope to have time to look at AZA stuff in June.

If there isn't a systemic problem, I don't see the need for anything that needs to be enforced at this point. It will just make it harder for the institutions that are already paying attention, and the ones that aren't will continue not too because I can't see any way to get teeth behind the enforcement.

I don't think you'll get an argument from anyone here about invert pain, or brain to body size or the need to keep these animals humanely.

I am not giving higher priority to bureaucratic concerns - that was someone else- but the more bureaucracy there is regarding an animal, the less likely it is to be displayed or researched. It's more of a practical thing - money, time and resources are real concerns in research and PA's. Is that a bad thing or a good thing - I'm not sure.

I am all for guidelines and standards, but am leery of anything that is 'enforceable' or 'legal' unless one can show that there is a real problem. I guess I am more interested in teaching than dictating.

 

[gjbarord]

Not sure how I missed this thread...

Greg

 

[Stavros]

In general, at least for me (and I'm going to carry out research on cephalopods for a living, so I know I'll be dealing with this, too), it seems kind of selfish to give a higher priority to bureaucratic concerns such as mountains of paperwork, rules that might 'backfire' (can you provide instances of when these sorts of things backfired?), advance permissions, or some claim that these directives get in the way of doing science than to the well-being of our captive cephalopods. These sort of claims - and I sincerely don't wish to offend, but this is quite true - are old hat when it comes to vertebrate welfare.

One reason I'll be visiting Euroceph is to find out as much as possible about the upcoming EU Directive and how it will affect current and future research. Having said this, I feel it's too early to visualize the impact (there's some time until the regulation is put into practice) other than obviously having less freedom to work with these animals. Aside from more paperwork, which is a headache, there will be changes in how much and what kind of projects will be funded. And the most important thing here is that we don't have conclusive evidence on nociception and pain. I think if there ever was a demand for more studies on this topic is now. That, and advancements on anaesthesia will help to make the best out of this.

On the point on intelligence and animals, I bet if you ask a bunch of researchers to give you a definition of "intelligence" you'll get as many answers. So it's obvious here that nothing is obvious when we talk about ceph intelligence. I find it very hard to compare molluscs with vertebrates. It's all about your perspective and how you filter this knowledge. You may call it intelligence but somebody else will say "complex behavior indeed, not necessarily intelligent."

 

[gjbarord]

Stavros,

Be on the lookout for something upcoming on cephalopod anesthesia/euthanasia...

To your point on defining intelligence, what is a complex behavior?

Greg

 

[Stavros]

Complex behavior can be defined as multiple strategies an organism uses to achieve a goal. based on available cues, one strategy can be more efficient than another. These strategies can be described as mechanisms,for example, the spiny lobster can return to home location from any point either by the use of landmarks, path integration, or by having a cognitive map and sense of orientation due to magnetoreception.

Looking forward for the article, is it on magnesium chloride or is it a new cocktail?

 

[neurobadger]

One reason I'll be visiting Euroceph is to find out as much as possible about the upcoming EU Directive and how it will affect current and future research. Having said this, I feel it's too early to visualize the impact (there's some time until the regulation is put into practice) other than obviously having less freedom to work with these animals. Aside from more paperwork, which is a headache, there will be changes in how much and what kind of projects will be funded. And the most important thing here is that we don't have conclusive evidence on nociception and pain. I think if there ever was a demand for more studies on this topic is now. That, and advancements on anaesthesia will help to make the best out of this.

On the point on intelligence and animals, I bet if you ask a bunch of researchers to give you a definition of "intelligence" you'll get as many answers. So it's obvious here that nothing is obvious when we talk about ceph intelligence. I find it very hard to compare molluscs with vertebrates. It's all about your perspective and how you filter this knowledge. You may call it intelligence but somebody else will say "complex behavior indeed, not necessarily intelligent."

I agree on your assessment of what complex behavior and intelligence are; admittedly, my approach is to give the cephalopod the benefit of the doubt just in case, but that is a personal preference in keeping with my own code of ethics.

On nociception and pain - what sort of conclusive evidence are you talking about? Conclusive evidence on how it works in cephs?

Why would the impact on cephalopod use be that much different than the impact on vertebrate use?

 

[Stavros]

I haven't mentioned anything about differences for the impact on vertebrates and invertebrates.

Your point on ethics is well taken, and I feel a lot of people share your approach. But the point here is that we can't enforce laws based on doubt alone. because we don't know about something. That's why I said before there's demand oforthese kind of studies. We need evidence based on physiological effects as well.

 

[robyn]

There is no such requirement for invertebrates (although I know of at least one case in the context of pain research, on tonmo user robyn's blog - http://www.tonmo.com/blog/entry.php?...ting-published.... - , where research has not been allowed to go forward because of animal welfare concerns about invertebrates.

Just to clarify, that was not the case nor is there any suggestion of it becoming so in future. The manuscript detailing the research was declined from one journal because the reveiwers objected to the focus and the conclusions of the study (suggesting we should have done a 'pain' study, rather than a nociceptive sensitisation study). That is a rational and reasonable decision for them to make - it's their journal and they have the right to define its purview.

As for regulations, I think some are needed, and if they are made they should be compulsory not voluntary. Regulations including some or all cephalopods are already operational in several countries and there is no reason that regulation, reasonably and responsibly implemented, need be onerous to the point of making research not worth doing. I support the use of regulations for many invertebrate species, since in almost all cases where a scientist has empirically considered 'does this injurious or noxious experience change the behaviour of this invertebrate in such a way that if a vertebrate behaved similarly, we would consider this evidence of pain or distress?' such evidence has been found. The lack of present regulation for invertebrates stems more from want of looking than negative findings.

As for the article neurobadger attached here, I do not think the discussion given in the section on octopus meets minimum referencing requirements and is almost purely anecdotal. The discussion on 'emotion' related to colour changes (red='angry') is needlessly anthropocentric and circular. Red could be aggressive, defensive, or aroused, but 'anger' is something in addition and I don't think their is sufficient evidence to support this claim. Defining white as fright and red as anger and then using this assertion as evidence for emotion is circular logic. The 'assessment of the whole situation' reported regarding change in tactics after anemone sting indicates nociception and learning, neither of which necessarily indicate pain or suffering. Nociception is not, in and of itself, pain.

While I'm critical of this particular paper, I greatly respect her experimental work and have had the pleasure of meeting and speaking with her at conferences, but this article is primarily an opinion piece and should be regarded as such.

 

[neurobadger]

Just to clarify, that was not the case nor is there any suggestion of it becoming so in future. The manuscript detailing the research was declined from one journal because the reveiwers objected to the focus and the conclusions of the study (suggesting we should have done a 'pain' study, rather than a nociceptive sensitisation study). That is a rational and reasonable decision for them to make - it's their journal and they have the right to define its purview.

Okay. Thanks for the clarification.

As for regulations, I think some are needed, and if they are made they should be compulsory not voluntary. Regulations including some or all cephalopods are already operational in several countries and there is no reason that regulation, reasonably and responsibly implemented, need be onerous to the point of making research not worth doing. I support the use of regulations for many invertebrate species, since in almost all cases where a scientist has empirically considered 'does this injurious or noxious experience change the behaviour of this invertebrate in such a way that if a vertebrate behaved similarly, we would consider this evidence of pain or distress?' such evidence has been found. The lack of present regulation for invertebrates stems more from want of looking than negative findings.

As for the article neurobadger attached here, I do not think the discussion given in the section on octopus meets minimum referencing requirements and is almost purely anecdotal. The discussion on 'emotion' related to colour changes (red='angry') is needlessly anthropocentric and circular. Red could be aggressive, defensive, or aroused, but 'anger' is something in addition and I don't think their is sufficient evidence to support this claim. Defining white as fright and red as anger and then using this assertion as evidence for emotion is circular logic. The 'assessment of the whole situation' reported regarding change in tactics after anemone sting indicates nociception and learning, neither of which necessarily indicate pain or suffering. Nociception is not, in and of itself, pain.

While I'm critical of this particular paper, I greatly respect her experimental work and have had the pleasure of meeting and speaking with her at conferences, but this article is primarily an opinion piece and should be regarded as such.

I agree that 'anger' is an anthropocentric overreach, but there does seem to be some evidence of alarm and aversive behavior - this is clearly not something the animal invites more of. I'm not sure, though, if there's some established standard by which one might consider 'this animal is clearly suffering' for invertebrates the same way there is for vertebrates.

What kinds of nociceptive receptors do cephalopods have? Are they much like the bare nerve endings of mammalian vertebrates?

Where do you draw the line between 'nociception' and 'pain'? I have sort of an idea, but what kinds of nociception would not constitute pain, irritation, or another aversive stimulus?

 

[robyn]

What kinds of nociceptive receptors do cephalopods have? Are they much like the bare nerve endings of mammalian vertebrates?

. No one know about cephalopods specifically, although currently my lab has a grant in review trying to get funding to look at this. We do know a lot about nociceptors in another mollusc, though, the neurosceince hero Aplysia, and if nociceptors are found in cephs its reasonable to expect them to be similar at the cellular, although not necessarily the circuit, level.
In Aplysia, nociceptor somata are somatotopically arranged in the pleural ganglion, which embryology suggests is homologous to the palliovisceral ganglion in cephalopods - a good place to start to look for nociceptors). Aplysia nociceptors, like all invert neurons, are unmyelinated. Mammalian nociceptors come in two classes, fast conducting myelinated A-delta fibres, which help mediate the reflexive withdrawal response, and C-fibres, which are unmyelinated and conduct slower pain impulses (so when you burn your finger on the stove, that quick jerk away is A-delta fibers in action, the few-seconds-later 'holy cr*p that hurt' response is your C-fibers in action.) Mammalian nociceptors also can and do fire spontaneously a long time after injury (which may have something to do with chronic pain syndromes), providing constant signals to the higher processing centers in the brain (thalamus, for example). It might well be that this spontaneous activity (that is, activity long after the cessation of the painful stimulus) is an important component of what 'pain' is - that ongoing negative reinforcement that reminds us not to be so stupid next time. Or not, no one really knows. However, we do know that spontaneous activity in nociceptors after noxious stimulation has never been observed in molluscs.

Where do you draw the line between 'nociception' and 'pain'?

Oh, this is a tough one. even the IASP (International Assn for the Study of Pain) gets a bit circular here. Currently I and a senior colleague have an article in press on the animal welfare implication of nociception in molluscs, and we go into the distinction mostly to point out its anthropcentrism and logical inconsistencies. I'm pasting an excert from that section here. The full paper should be out sometime this summer in the Natl Academy's animal welfare journal.

Nociception is a capacity to react to tissue damage or impending damage with activation of sensory pathways, with or without conscious sensation. Activity in nociceptive sensory pathways usually results in reflexive behavioral responses and may or may not result in other responses. Reflexive withdrawal responses tend to be mediated by very simple sensory-motor circuits optimized for speed and reliability, and can occur without input from higher processing centers, although more complex escape and avoidance behaviors involve more complex neural circuits (Walters, 1994; Chase, 2002). Even in humans the initial reflexive response to a noxious stimulus is sometimes faster than can be consciously perceived, and nociceptors can sometimes be activated without conscious sensation (Adriaensen et al., 1980). Invertebrates that lack appropriate processing centers may only be capable of this rapid, unconscious processing.
The definition of pain widely accepted by scientific investigators is “an unpleasant sensory and emotional experience associated with actual or potential tissue damage , or described in terms of such damage” (Merskey and Bogduk, 1994). The emotional component required by this definition of pain makes its identification in other species, and especially in species quite different from humans, extremely difficult, if not impossible. This is because emotion is usually defined in terms of conscious experience (e.g., Izard, 2009), and while evidence of consciousness in some animals is available, proof of consciousness is not (e.g., Allen, 2004). It is therefore important to draw a distinction between nociception as detection of a noxious stimulus (which can be recognized scientifically by unambiguous behavioral and neural responses), and pain as the unpleasant feeling associated with that stimulus (with the feeling inferred by behavioral and neural responses of uncertain relation to consciousness). Moreover, this emotional response during pain may be linked to cognition, “knowing” in some sense that the sensation is negative and involves a threat to the body. Whereas nociception leading to a nociceptive response can be mediated by the simplest of neural circuits, in principle just a single nociceptor connected to an effector system (e.g., a muscle), pain requires neural circuitry that incorporates additional functions, some of which might entail highly complex processing by very large numbers of neurons.

And that is a 'wall o' text' to do Monty proud. He'd be having a field day in this thread, I think!

 

[Thales]

I miss Monty.

 

[neurobadger]

. No one know about cephalopods specifically, although currently my lab has a grant in review trying to get funding to look at this. We do know a lot about nociceptors in another mollusc, though, the neurosceince hero Aplysia, and if nociceptors are found in cephs its reasonable to expect them to be similar at the cellular, although not necessarily the circuit, level.
In Aplysia, nociceptor somata are somatotopically arranged in the pleural ganglion, which embryology suggests is homologous to the palliovisceral ganglion in cephalopods - a good place to start to look for nociceptors). Aplysia nociceptors, like all invert neurons, are unmyelinated. Mammalian nociceptors come in two classes, fast conducting myelinated A-delta fibres, which help mediate the reflexive withdrawal response, and C-fibres, which are unmyelinated and conduct slower pain impulses (so when you burn your finger on the stove, that quick jerk away is A-delta fibers in action, the few-seconds-later 'holy cr*p that hurt' response is your C-fibers in action.) Mammalian nociceptors also can and do fire spontaneously a long time after injury (which may have something to do with chronic pain syndromes), providing constant signals to the higher processing centers in the brain (thalamus, for example). It might well be that this spontaneous activity (that is, activity long after the cessation of the painful stimulus) is an important component of what 'pain' is - that ongoing negative reinforcement that reminds us not to be so stupid next time. Or not, no one really knows. However, we do know that spontaneous activity in nociceptors after noxious stimulation has never been observed in molluscs.

What I'm not aware of is whether spontaneous activity of nociceptors is a normal (i.e. not associated with chronic pain or any other neurological pathology). Is it also associated with the normal experience of pain?

Even in humans the initial reflexive response to a noxious stimulus is sometimes faster than can be consciously perceived, and nociceptors can sometimes be activated without conscious sensation (Adriaensen et al., 1980). Invertebrates that lack appropriate processing centers may only be capable of this rapid, unconscious processing. ... Whereas nociception leading to a nociceptive response can be mediated by the simplest of neural circuits, in principle just a single nociceptor connected to an effector system (e.g., a muscle), pain requires neural circuitry that incorporates additional functions, some of which might entail highly complex processing by very large numbers of neurons.

How would we be able to tell if any species of invertebrate had these processing centers? We seem to regard most nonhuman vertebrates as having these processing centers.

I've definitely seen a number of electrophysiological and cell-biology studies bearing out some possibilities in this area (I could be totally wrong in my interpretation of the studies, of course) as regards Octopus vulgaris, at least.

I emailed the folks responsible for the EU directive and asked them if they had a list of sources they used for their decision; this is what they sent me.

Dear (neurobadger),

The decision to include cephalopods was the result of the compromise between the Commission, the Council and the European Parliament. The original Commission proposal was based on a Scientific Opinion that was tasked specifically among other issues to examine the sentience of different species and their possible inclusion under the scope of the Directive.

You can find the Scientific Opinion at http://ec.europa.eu/environment/chemicals/lab_animals/scientific_en.htm The references should be found at the end of the Scientific Report;

the original Commission proposal at http://ec.europa.eu/environment/chemicals/lab_animals/proposal_en.htm

and the final adopted Directive 2010/63/EU at http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=OJ:L:2010:276:0033:0079:EN:PDF

With kind regards,

Your team for the Protection of Experimental Animals
DG Environment
European Commission

 

[neurobadger]

The Scientific Report is at http://ec.europa.eu/environment/chemicals/lab_animals/pdf/efsa_scientificreport_en.pdf and the background on cephalopods starts on page 15.

 

[neurobadger]

One bit I found especially germane to this discussion:

2.3.3. Nociception and Pain in Invertebrates
Smith and Boyd (1991) suggested seven criteria indicating the capacity for the
experience of pain in non-human animals and these are presented in slightly
modified form below.
1. Possession of receptors sensitive to noxious stimuli, located in functionally useful
positions on or in the body, and connected by nervous pathways to the lower parts
of a central nervous system
2. Possession of brain centres which are higher in the sense of level of integration of
brain processing (especially a structure analogous to the human cerebral cortex).
3. Possession of nervous pathways connecting the nociceptive system to the higher
brain centres.
4. Receptors for opioid substances found in the central nervous system, especially the
brain.
5. Analgesics modify an animal's response to stimuli that would be painful for a
human.
6. An animal's response to stimuli that would be painful for a human is functionally
similar to the human response (that is, the animal responds so as to avoid or
minimise damage to its body).
7. An animal's behavioural response persists and it shows an unwillingness to
resubmit to a painful procedure; the animal can learn to associate apparently nonpainful
with apparently painful events

p. 27, an extended discussion is following

Are we holding invertebrates to the same standards that we hold vertebrates as regards our judgment of their capacity for pain? If not, how good is our justification for it?

I am aware how point 2 of this might be difficult, and point 3 - well, last I heard the jury's out on how well octopus arm sensation is integrated with the central nervous system - but I don't think the other five points are very contestable.

 

[robyn]

What I'm not aware of is whether spontaneous activity of nociceptors is a normal (i.e. not associated with chronic pain or any other neurological pathology). Is it also associated with the normal experience of pain?

Yes and no, and I should have been clearer about what 'spontaneous' means - there appears to be a low proportion of nociceptors (between ~5 -15%) that will fire truly spontaneously in culture, in isolation from extrinsic signals and not having previously been primed by injury or painful stimulation, its hard to tell for sure if this follows in vivo, but most likely it does.

Nociceptors do fire repeatedly in response to pain, but since this is at the time of the tissue insult or in response to its immediate effects (inflammation, for example), its not strictly speaking, spontaneous, if we define spontaneous as an intrinsic property of the neuron itself and not being driven by any extrinsic factors. In cells that experience either direct injury or are exposed to injury or pain-related signals, the incidence of repeated firing without additional stimulation is much higher, but this is not spontaneous in the true sense. Threshold for firing decreases and firing intensity increases when a previously injured region is stimulated in a way that would not normally be painful, and this is interpreted as pain. When the neurons continue to fire after the injury is healed and there is no need for pain signals to persist, this is usually considered pathological spontaneous activity.

In inverts, nociceptors fire in response to immediate noxious stimulation and this firing patter can last minutes, but there does not appear to be ongoing firing after pain or injury without additional stimulation (like poking at bruised tissue).

The numbered list from EU directive is very interesting and looks about perfect to me except that I disagree with 4. Opiods are not the only endogenous analgesics, and the evidence for their presence, at least in molluscs, is not conclusive.