In 2010, an octopus named Paul gained worldwide fame by predicting winners for the World Cup games with impressive accuracy by choosing a snack from one of two boxes labeled with the competing team flags [1]. While Paul was not operating on a deep understanding of soccer, octopuses, and cephalopods in general, have proven themselves to be remarkably intelligent creatures. They have been studied in the past to understand their behavior and breeding habits (as they are an important source of food for humans and oceanic predators alike), but are also emerging as a model organism for cognition and evolution of intelligence.
Squid were used as early as the 1970s in some of the original experiments to determine the electrical properties of neurons [2]. Studies had previously been done to determine how cell membranes conduct electrical impulses, with confusing results. Enter the squid. In order to control quick movements, squid have nerve cells with giant axons. This part of the nerve was useful for electrical experiments because of its large size and allowed for more precise measurements with the equipment that was available in the 1970s. Other early research mostly focused on behavior and aquaculture. Given that humans fish over two million tons of cephalopods every year, a lot of research is focused on understanding how various ocean environments affect cephalopod populations [3]. This area of research aims to better predict population sizes from year to year, in order to avoid overfishing.
Recently, the field of cephalopod research has spread in new directions. Since 2006, every category of aquaculture, behavior, climate change, cognition, genetics, neuroscience, and welfare had at least 10 papers published, and the largest category, behavior, saw over 450 papers published [4]. This past November, cephalopod scientists from all over the world gathered for the the triennial Cephalopod International Advisory Council (CIAC) Conference. Many of the talks at the CIAC conference focused on taxonomy, ecosystem roles, and other fields of study that only require working with wild-caught, dead samples of cephalopods, pulled up from trawling nets. Even studying behavior can be done by simply observing living cephalopods for their natural life span, without invasive procedures. Studying development or neurology of cephalopods, however, requires procedures or euthanasia that have the potential to cause serious pain and distress to the animal.
As with all research on live subjects, the question of ethics and animal treatment must be addressed. Invertebrates typically have a small brain size and low neuron number compared to their body size, and most lab invertebrates are very small, like fruit flies or nematodes. Because of this, lab animals have largely been separated into vertebrates and invertebrates, each group with its own protections, or lack thereof. As early as the 1870s, vertebrate animals have had regulations on their care and use as laboratory animals [5]. Currently, in order to use live vertebrate animals in research, a protocol detailing exactly what procedures will be done, how pain and suffering is minimized, and a justification for using live vertebrates instead of another system must be submitted. In the United States, this protocol has to be approved by an Institutional Care and Use Committee (or IACUC). The goal of these regulations, which exist in various forms around the world, is to minimize harm to the animal while still allowing for their use as research subjects.
Since cephalopods are invertebrates, in the United States they are guaranteed almost no protections. However, they have the largest invertebrate brains, both in size and number of neurons, and are remarkably intelligent, and show distress, pain, and lasting damage [6]. For example, cephalopods avoid painful stimuli, such as electric shock, and guard wounds with their arms or tentacles [7]. Since invertebrates are excluded from the US Animal Welfare Act, the decision to review protocols on invertebrate research is up to the discretion of the IACUC at a particular institution to decide if and how invertebrate protocols will be evaluated. Some institutions may require a similar level of review for cephalopods as for vertebrate animals, while others may choose not to review any invertebrate protocols at all [8]. Columbia University, for example, falls somewhere in the middle. According to the head of Columbia’s IACUC, a cephalopod researcher would have to discuss protocols with a staff veterinarian and the IACUC, but would not be required to submit a formal protocol for IACUC review and approval.
Not all cephalopod research is so unregulated. In 2013, the European Union implemented EU Directive 2010/63/EU, which granted protections previously reserved for vertebrate species to organisms in the Class Cephalopoda. Previously, one species, Octopus vulgaris, had been granted these protections. Based on the scientific evidence that cephalopods experience pain, suffering, distress, and lasting harm, the directive extended these protections to all cephalopods [5]. An extensive review of care guidelines was published in 2015, providing a basis for researchers to establish ethical cephalopod studies [7]. Similarly, Canada’s Council on Animal Care guidelines, like EU Directive 2010/63/EU, requires protocols to be submitted for cephalopods and other higher invertebrates.
Unlike the EU, it’s unclear why the United States hasn’t addressed the issue of regulating cephalopod research. It’s possible that, since cephalopods are a relatively new model organism for behavior and neurology research, the issue simply isn’t large enough to address yet. There are very few cephalopod species that have a closed life cycle, meaning that the organism can be raised entirely within the lab without requiring wild animal capture at any life stage. With advances in cephalopod aquaculture, however, the Marine Biological Laboratory, a leading center for marine research in Massachusetts, now breeds several types of cephalopod that are available for request [9]. However, they have yet to establish any animal treatment guidelines to follow for the labs that request eggs or animals.
It remains to be seen whether the expansion of aquaculture and closing of life cycles, or simply the wider understanding of the intelligence of cephalopods, will result in a nationwide change in cephalopod regulations in the United States. For now, each lab or institution must plunge into the deep and figure out how to navigate cephalopod research on its own.
References
1. Kelsey, E. Don't Mess with the Octopus: Oracle Paul Celebrates Perfect World Cup Record. Retrieved from http://www.spiegel.de. (July 12th, 2010)
2. Cole, K. S. Electrical properties of the squid axon sheath. Biophys J 16, 137–142 (1976).
3. Hempel, E. Cephalopods. GLOBEFISH Highlights 430-40 (2018).
4. O’Brien, C. E., Roumbedakis, K. & Winkelmann, I. E. The Current State of Cephalopod Science and Perspectives on the Most Critical Challenges Ahead From Three Early-Career Researchers. Front Physiol 9, 700 (2018).
5. Finn, M.A; Stark, J.F. Medical science and the Cruelty to Animals Act 1876: A re-examination of anti-vivisectionism in provincial Britain. Stud Hist Philos Biol Biomed Sci 49, 12-23 (2015).
6. EFSA Panel on Animal Health and Welfare. Opinion of the Scientific Panel on Animal Health and Welfare (AHAW) on a request from the Commission related to the ‘Aspects of the biology and welfare of animals used for experimental and other scientific purposes’. EFSA J 2005; 292: 1–136.
7. Fiorito, G. et al. Guidelines for the Care and Welfare of Cephalopods in Research -A consensus based on an initiative by CephRes, FELASA and the Boyd Group. Lab. Anim. 49, 1–90 (2015).
8. Harvey-Clark, C. IACUC Challenges in Invertebrate Research. ILAR J 52, 213–220 (2011).
9. http://www.mbl.edu/cephalopod-program/
