The Long Road from Religious and Ethical Traditions to Welfare of Invertebrates.

Ethical behaviour tends to lead to the welfare consideration of animals, but much less so for invertebrates. Indigenous tradition often valued all animals as having an important role in life on the planet, a practical application of modern ecology. The Judaeo-Christian-Islamic tradition postulated 'man' as having dominion over all of Earth, resulting in anthropocentrism and careless practices. In contrast, the Buddhist/Hindu belief in rebirth leads to ahisma, or doing no harm. In the face of capitalist systems, practice does not necessarily follow these beliefs, especially in the 'shepherding' of domestic animals. Only Jainist beliefs value the lives of all invertebrates. Philosophers are often divorced from the physiological reality of the animals they muse about, and science's traditions of objectivity and the simplest possible explanation of behaviour led to ignorance of invertebrates' abilities. Ninety-seven percent of animals on the planet are invertebrates. We have a long way to go to provide moral standing and welfare consideration, which is consistent with the new information about the sentience of some of these animals.

Ethics and Invertebrates: The Problem Is Us.

In the last few decades, science has begun to make great strides at understanding how varied, fascinating, and intelligent invertebrate animals are. Because they are poorly known, the invertebrates that make up about 98% of the animals on the planet have been overlooked. Because they are seen as both simple and unattractive, children and their teachers, as well as the general public, do not think they need care. Because until recently we did not know they can be both intelligent and sensitive-bees can learn from each other, butterflies can navigate huge distances, octopuses are smart, and lobsters can feel pain-we have to give them the consideration they deserve. This collection of papers should help us to see how the lives of invertebrates are tightly linked to ours, how they live, and what they need in terms of our consideration and care.

Why Are Octopuses Going to Be the 'Poster Child' for Invertebrate Welfare?

Animal welfare consideration and actions are generally addressed to animals similar to us, predominantly large mammals. Invertebrates are neglected partly because they are unknown, though new exploration of the oceans has helped with this. Also, we know little about their ecology and welfare. This is gradually changing, and the octopuses are likely to be the first beneficiaries. Scientists are finding that cephalopods are far more intelligent than we thought, with the Cambridge Declaration of Consciousness suggesting they might possess this quality of mind. Partly as a result, the European Union has described and demanded good care for cephalopods such as the octopus in captivity. Public opinion has been swayed to approval by anecdotes of octopuses doing unusual actions, and by several recent books pointing out interesting and intelligent behavior of cephalopods. Aquariums have begun to feature octopuses for them. With this progress, welfare of invertebrate animals has begun to matter. While the octopuses will be the first animal group to benefit, they may pave the way for us to see that different does not mean unworthy of regard and welfare consideration.

Ethics and Care: For Animals, Not Just Mammals.

In the last few decades, we have made great strides in recognizing ethics and providing care for animals, but the focus has been mainly on mammals. This stems from a bias of attention not only in research but predominantly in non-scientists' attention (to 'popular' animals), resulting partly from discussion about and depiction of animals in publications addressed to the public. This is somewhat due to political pressure, and can result in uneven conservation efforts and biases in targets for welfare concerns. As a result, there has been a huge backlash again, with concerns about pain sensitivity and welfare in fish, and a less focused but more pervasive omission of consideration of all invertebrates. That means welfare efforts are focused on 0.2% of the animal species on the planet, and education about non-mammals, particularly addressed to children, is necessary to broaden this focus and care more fully for the inhabitants of the planet.

Male Alternative Reproductive Tactics and Associated Evolution of Anatomical Characteristics in Loliginid Squid.

Loliginid squids provide a unique model system to explore male alternative reproductive tactics (ARTs) and their linkage to size, behavioral decision making, and possibly age. Large individuals fight one another and the winners form temporary consortships with females, while smaller individuals do not engage in male-male agonistic bouts but use various sneaker tactics to obtain matings, each with varying mating and fertilization success. There is substantial behavioral flexibility in most species, as smaller males can facultatively switch to the alternative consort behaviors as the behavioral context changes. These forms of ARTs can involve different: mating posture; site of spermatophore deposition; fertilization success; and sperm traits. Most of the traits of male dimorphism (both anatomical and behavioral) are consistent with traditional sexual selection theory, while others have unique features that may have evolved in response to the fertilization environment faced by each temporary or permanent male morph.

What is cognition?

Eleven authors with disparate relevant backgrounds give their view on what is meant by the word "cognition".

Effects of Friend Virus Infection and Regulatory T Cells on the Antigen Presentation Function of B Cells.

Friend virus (FV) is a naturally occurring mouse retrovirus that infects dividing cells of the hematopoietic lineage, including antigen-presenting cells (APCs). The infection of APCs by viruses often induces their dysfunction, and it has been shown that FV infection reduces the ability of dendritic cells (DCs) to prime critical CD8+ T cell responses. Nonetheless, mice mount vigorous CD8+ T cell responses, so we investigated whether B cells might serve as alternative APCs during FV infection. Direct ex vivo analysis of B cells from FV-infected mice revealed that infected but not uninfected B cells upregulated expression of the costimulatory molecules CD80, CD86, and CD40, as well as major histocompatibility complex class II (MHC-II) molecules. Furthermore, in vitro studies showed that, compared to uninfected B cells from the same mice, the FV-infected B cells had significantly enhanced APC function, as measured by their capacity to prime CD8+ T cell activation and proliferation. Thus, in contrast to DCs, infection of B cells with FV enhanced their APC capacity and ability to stimulate the CD8+ T cell responses essential for virus control. FV infections also induce the activation and expansion of regulatory T cells (Tregs), so it was of interest to determine the impact of Tregs on B cell activation. The upregulation of costimulatory molecule expression and APC function of B cells was even more strongly enhanced by in vivo depletion of regulatory T cells than infection. Thus, Tregs exert potent homeostatic suppression of B cell activation that is partially overcome by FV infection.IMPORTANCE The primary role of B cells in immunity is considered the production of pathogen-specific antibodies, but another, less-well-studied, function of B cells is to present foreign antigens to T cells to stimulate their activation and proliferation. Dendritic cells (DCs) are considered the most important antigen-presenting cells (APCs) for CD8+ T cells, but DCs lose APC function when infected with Friend virus (FV), a model retrovirus of mice. Interestingly, B cells were better able to stimulate CD8+ T cell responses when they were infected with FV. We also found that the activation status of B cells under homeostatic conditions was potently modulated by regulatory T cells. This study illustrates an important link between B cell and T cell responses and illustrates an additional mechanism by which regulatory T cells suppress critical T cell responses during viral infections.

Variability in the "stereotyped" prey capture sequence of male cuttlefish (Sepia officinalis) could relate to personality differences.

Studies of animal personality have shown consistent between-individual variation in behaviour in many social and non-social contexts, but hunting behaviour has been overlooked. Prey capture sequences, especially in invertebrates, are supposed to be quite invariant. In cuttlefish, the attack includes three components: attention, positioning, and seizure. The previous studies indicated some variability in these components and we quantified it under the hypothesis that it could relate to personality differences. We, therefore, analysed predation sequences of adult cuttlefish to test their association with personality traits in different contexts. Nineteen subjects were first exposed to an "alert" and a "threat" test and then given a live prey, for 10 days. Predation sequences were scored for components of the attack, locomotor and postural elements, body patterns, and number of successful tentacle ejections (i.e. seizure). PCA analysis of predatory patterns identified three dimensions accounting for 53.1%, 15.9%, and 9.6% of the variance and discriminating individuals based on "speed in catching prey", "duration of attack behaviour", and "attention to prey". Predation rate, success rate, and hunting time were significantly correlated with the first, second, and third PCA factors, respectively. Significant correlations between capture patterns and responsiveness in the alert and threat tests were found, highlighting a consistency of prey capture patterns with measures of personality in other contexts. Personality may permeate even those behaviour patterns that appear relatively invariant.

B Cell Requirement for Robust Regulatory T Cell Responses to Friend Retrovirus Infection.

Regulatory T cells (Tregs) are immunosuppressive cells of the immune system that control autoimmune reactivity. Tregs also respond during immune reactions to infectious agents in order to limit immunopathological damage from potent effectors such as CD8+ cytolytic T lymphocytes. We have used the Friend virus (FV) model of retroviral infection in mice to investigate how viral infections induce Tregs. During acute FV infection, there is significant activation and expansion of thymus-derived (natural) Tregs that suppress virus-specific CD8+ T cell responses. Unlike conventional T cells, the responding Tregs are not virus specific, so the mechanisms that induce their expansion are of great interest. We now show that B cells provide essential signals for Treg expansion during FV infection. Treg responses are greatly diminished in B cell-deficient mice but can be restored by adoptive transfers of B cells at the time of infection. The feeble Treg responses in B cell-deficient mice are associated with enhanced virus-specific CD8+ T cell responses and accelerated virus control during the first 2 weeks of infection. In vitro experiments demonstrated that B cells promote Treg activation and proliferation through a glucocorticoid-induced receptor superfamily member 18 (GITR) ligand-dependent mechanism. Thus, B cells play paradoxically opposing roles during FV infection. They provide proliferative signals to immunsosuppressive Tregs, which slows early virus control, and they also produce virus-specific antibodies, which are essential for long-term virus control.IMPORTANCE When infectious agents invade a host, numerous immunological mechanisms are deployed to limit their replication, neutralize their spread, and destroy the host cells harboring the infection. Since immune responses also have a strong capacity to damage host cells and tissues, their magnitude, potency, and duration are under regulatory control. Regulatory T cells are an important component of this control, and the mechanisms that induce them to respond and exert immunosuppressive regulation are of great interest. In the current report, we show that B cells, the cells responsible for making pathogen-specific antibodies, are also involved in promoting the expansion of regulatory T cells during a retroviral infection. In vitro studies demonstrated that they do so via stimulation of the Tregs through interactions between cell surface molecules: GITR interactions with its ligand (GITRL) on B cells and GITR on regulatory T cells. These findings point the way toward therapeutics to better treat infections and autoimmune diseases.

An ethogram for Benthic Octopods (Cephalopoda: Octopodidae).

The present paper constructs a general ethogram for the actions of the flexible body as well as the skin displays of octopuses in the family Octopodidae. The actions of 6 sets of structures (mantle-funnel, arms, sucker-stalk, skin-web, head, and mouth) combine to produce behavioral units that involve positioning of parts leading to postures such as the flamboyant, movements of parts of the animal with relation to itself including head bob and grooming, and movements of the whole animal by both jetting in the water and crawling along the substrate. Muscular actions result in 4 key changes in skin display: (a) chromatophore expansion, (b) chromatophore contraction resulting in appearance of reflective colors such as iridophores and leucophores, (c) erection of papillae on the skin, and (d) overall postures of arms and mantle controlled by actions of the octopus muscular hydrostat. They produce appearances, including excellent camouflage, moving passing cloud and iridescent blue rings, with only a few known species-specific male visual sexual displays. Commonalities across the family suggest that, despite having flexible muscular hydrostat movement systems producing several behavioral units, simplicity of production may underlie the complexity of movement and appearance. This systematic framework allows researchers to take the next step in modeling how such diversity can be a combination of just a few variables. (PsycINFO Database Record

Transitions during cephalopod life history: the role of habitat, environment, functional morphology and behaviour.

Cephalopod life cycles generally share a set of stages that take place in different habitats and are adapted to specific, though variable, environmental conditions. Throughout the lifespan, individuals undertake a series of brief transitions from one stage to the next. Four transitions were identified: fertilisation of eggs to their release from the female (1), from eggs to paralarvae (2), from paralarvae to subadults (3) and from subadults to adults (4). An analysis of each transition identified that the changes can be radical (i.e. involving a range of morphological, physiological and behavioural phenomena and shifts in habitats) and critical (i.e. depending on environmental conditions essential for cohort survival). This analysis underlines that transitions from eggs to paralarvae (2) and from paralarvae to subadults (3) present major risk of mortality, while changes in the other transitions can have evolutionary significance. This synthesis suggests that more accurate evaluation of the sensitivity of cephalopod populations to environmental variation could be achieved by taking into account the ontogeny of the organisms. The comparison of most described species advocates for studies linking development and ecology in this particular group.

Marine invertebrates: communities at risk.

Our definition of the word 'animal' centers on vertebrates, yet 99% of the animals on the planet are invertebrates, about which we know little. In addition, although the Census of Marine Life (COML.org) has recently conducted an extensive audit of marine ecosystems, we still do not understand much about the animals of the seas. Surveys of the best-known ecosystems, in which invertebrate populations often play a key role, show that the invertebrate populations are affected by human impact. Coral animals are the foundation of coral reef systems, which are estimated to contain 30% of the species in the ocean. Physical impact and chemical changes on the water severely damage these reefs, and may lead to the removal of these important habitats. Tiny pteropod molluscs live in huge numbers in the polar seas, and their fragile shells are particularly vulnerable to ocean acidification. Their removal would mean that fishes on which we depend would have a hugely diminished food supply. In the North Sea, warming is leading to replacement of colder water copepods by warmer water species which contain less fat. This is having an effect on the birds which eat them, who enrich the otherwise poor land on which they nest. Conversely, the warming of the water and the loss of top predators such as whales and sharks has led to an explosion of the jumbo squid of the Pacific coast of North America. This is positive in the development of a squid fishery, yet negative because the squid eat fish that have been the mainstay of the fishery along that coast. These examples show how invertebrates are key in the oceans, and what might happen when global changes impact them.

Philosophical background of attitudes toward and treatment of invertebrates.

People who interact with or make decisions about invertebrate animals have an attitude toward them, although they may not have consciously worked it out. Three philosophical approaches underlie this attitude. The first is the contractarian, which basically contends that animals are only automata and that we humans need not concern ourselves with their welfare except for our own good, because cruelty and neglect demean us. A second approach is the utilitarian, which focuses on gains versus losses in interactions between animals, including humans. Given the sheer numbers of invertebrates-they constitute 99% of the animals on the planet- this attitude implicitly requires concern for them and consideration in particular of whether they can feel pain. Third is the rights-based approach, which focuses on humans-treatment of animals by calling for an assessment of their quality of life in each human-animal interaction. Here scholars debate to what extent different animals have self-awareness or even consciousness, which may dictate our treatment of them. Regardless of the philosophical approach to invertebrates, information and education about their lives are critical to an understanding of how humans ought to treat them.

Octopuses (Enteroctopus dofleini) recognize individual humans.

This study exposed 8 Enteroctopus dofleini separately to 2 unfamiliar individual humans over a 2-week period under differing circumstances. One person consistently fed the octopuses and the other touched them with a bristly stick. Each human recorded octopus body patterns, behaviors, and respiration rates directly after each treatment. At the end of 2 weeks, a body pattern (a dark Eyebar) and 2 behaviors (reaching arms toward or away from the tester and funnel direction) were significantly different in response to the 2 humans. The respiration rate of the 4 larger octopuses changed significantly in response to the 2 treatments; however, there was no significant difference in the 4 smaller octopuses' respiration. Octopuses' ability to recognize humans enlarges our knowledge of the perceptual ability of this nonhuman animal, which depends heavily on learning in response to visual information. Any training paradigm should take such individual recognition into consideration as it could significantly alter the octopuses' responses.

Evaluation of phospholipid and liposomal S-adenosyl methionine for the treatment of liver injury in a murine model.

We have used a murine model of Acetaminophen induced hepatoxicity to determine if S-adenosyl methionine 1,4 butanedisulfonate (SD4) in liposomes can prevent liver injury when administered immediately prior to acetaminophen, as judged by serum aspartate aminotransferase and alanine aminotransferase levels, and histological evidence of liver necrosis. No protection was observed when mice received 1 g/kg unencapsulated SD4. Partial protection was observed with 5 or 0.5 mg/kg SD4 in unextruded distearoylphosphatidylglycerol (DSPG) liposomes. Protection comparable to that seen in mice receiving encapsulated SD4 is achieved when mice received lipid alone in equivalent amounts, suggesting that the contribution of encapsulated SD4 to the efficacy of the liposomes may be minimal. Unextruded distearoylphosphatidylcholine (DSPC) liposomes show only slight effects even at 50 mg/kg SD4. This is likely caused by the size of unextruded DSPC lipsomes, because extruded DSPC liposomes, whose size is smaller, are of comparable efficacy to unextruded DSPG liposomes.

Cephalopod consciousness: behavioural evidence.

Behavioural evidence suggests that cephalopod molluscs may have a form of primary consciousness. First, the linkage of brain to behaviour seen in lateralization, sleep and through a developmental context is similar to that of mammals and birds. Second, cephalopods, especially octopuses, are heavily dependent on learning in response to both visual and tactile cues, and may have domain generality and form simple concepts. Third, these animals are aware of their position, both within themselves and in larger space, including having a working memory of foraging areas in the recent past. Thus if using a 'global workspace' which evaluates memory input and focuses attention is the criterion, cephalopods appear to have primary consciousness.

The packaging problem: bivalve prey selection and prey entry techniques of the octopus Enteroctopus dofleini.

Many predators face a complex step of prey preparation before consumption. Octopuses faced with bivalve prey use several techniques to penetrate the shells to gain access to the meat inside. When given prey of mussels Mytilus trossulus, Manila clams Venerupis philippinarum, and littleneck clams Protothaca staminea, Enteroctopus dofleini solved the problem differently. They pulled apart V. philippinarum and M. trossulus, which had the thinnest shells and the least pulling resistance. P. staminea were eaten after the shells had been chipped or had been penetrated by drilling, presumably to inject a toxin. Likely because of these differences, octopuses consumed more V. philippinarum and M. trossulus than P. staminea when the mollusks were given to them either 1 species at a time or all together. However, when the shells were separated and the penetration problem removed, the octopuses predominantly chose P. staminea and nearly ignored M. trossulus. When V. philippinarum were wired shut, octopuses switched techniques. These results emphasize that octopuses can learn on the basis of nonvisual information and monitor their body position to carry out feeding actions.

Ethics and invertebrates: a cephalopod perspective.

This paper first explores 3 philosophical bases for attitudes to invertebrates, Contractarian/Kantian, Utilitarian, and Rights-based, and what they lead us to conclude about how we use and care for these animals. We next discuss the problems of evaluating pain and suffering in invertebrates, pointing out that physiological responses to stress are widely similar across the animal kingdom and that most animals show behavioral responses to potentially painful stimuli. Since cephalopods are often used as a test group for consideration of pain, distress and proper conditions for captivity and handling, we evaluate their behavioral and cognitive capacities. Given these capacities, we then discuss practical issues: minimization of their pain and suffering during harvesting for food; ensuring that captive cephalopods are properly cared for, stimulated and allowed to live as full a life as possible; and, lastly, working for their conservation.

When do octopuses play? Effects of repeated testing, object type, age, and food deprivation on object play in Octopus vulgaris.

Studying play behavior in octopuses is an important step toward understanding the phylogenetic origins and function of play as well as the cognitive abilities of invertebrates. Fourteen Octopus vulgaris (7 subadults and 7 adults) were presented 2 Lego objects and 2 different food items on 7 consecutive days under 2 different levels of food deprivation. Nine subjects showed play-like behavior with the Lego objects. There was no significant difference in play-like behavior corresponding to food deprivation, age, and sex of the octopuses. The sequence of behaviors, from exploration to play-like behavior, had a significant influence on the establishment of play-like behavior, as it occurred mostly on Days 3-6 of the 7-day experiment. The pattern of development of play-like activities after a period of exploration and habituation in this study agrees with the hypothesis that object play follows object exploration. A homologous origin of this behavioral trait in vertebrates and invertebrates is highly unlikely, as the last common ancestor might not have had the cognitive capacity to possess this trait.