Inspiration, simulation and design for smart robot manipulators from the sucker actuation mechanism of cephalopods

Octopus arms house 200-300 independently controlled suckers that can alternately afford an octopus fine manipulation of small objects and produce high adhesion forces on virtually any non-porous surface. Octopuses use their suckers to grasp, rotate and reposition soft objects (e.g., octopus eggs) without damaging them and to provide strong, reversible adhesion forces to anchor the octopus to hard substrates (e.g., rock) during wave surge. The biological 'design' of the sucker system is understood to be divided anatomically into three functional groups: the infundibulum that produces a surface seal that conforms to arbitrary surface geometry; the acetabulum that generates negative pressures for adhesion; and the extrinsic muscles that allow adhered surfaces to be rotated relative to the arm. The effector underlying these abilities is the muscular hydrostat. Guided by sensory input, the thousands of muscle fibers within the muscular hydrostats of the sucker act in coordination to provide stiffness or force when and where needed. The mechanical malleability of octopus suckers, the interdigitated arrangement of their muscle fibers and the flexible interconnections of its parts make direct studies of their control challenging. We developed a dynamic simulator (ABSAMS) that models the general functioning of muscular hydrostat systems built from assemblies of biologically constrained muscular hydrostat models. We report here on simulation studies of octopus-inspired and artificial suckers implemented in this system. These simulations reproduce aspects of octopus sucker performance and squid tentacle extension. Simulations run with these models using parameters from man-made actuators and materials can serve as tools for designing soft robotic implementations of man-made artificial suckers and soft manipulators.     

Assessing <Emphasis Type="Italic">Octopus vulgaris</Emphasis> distribution using presence-only model methods

The distribution of the common octopus (Octopus vulgaris) in the Mediterranean and Eastern Atlantic waters is evaluated using two presence-only analyses: The maximum entropy model (Maxent) and the ecological niche factor analysis (ENFA). Maxent predicts those geographical areas that satisfy the environmental or abiotic requirements of a species while ENFA explores the niche and habitat preferences of O. vulgaris. The analyses were implemented recovering the spatial information from 213 octopus presence data collected from surveys and bibliographical records. Together, these analyses provided reasonable estimates of the species distribution and the octopus habitat. Among the gathered set of explanatory environmental variables, sea bottom temperature, sea bottom salinity, surface dissolved oxygen and sea surface chlorophyll-α appear as the main variables involved in O. vulgaris distribution. These results were confronted with the available literature.     

Gonadal development and reproductive strategies of the tropical octopus (Octopus insularis) in northeast Brazil

Octopus insularis inhabits shallow waters along the coast and oceanic islands of northeastern Brazil, where it is the main target of commercial fishing of octopuses. This study aims to investigate the hypothesis that the tropical O. insularis has a distinct gonadal development and reproductive features when compare to its congener O. vulgaris from the subtropical regions. In order to describe its reproductive development, 545 octopuses were collected in the Northeastern Brazil. A good correspondence was observed between the gonad morphology and its histological structure. Oocytes in different development stages were observed in mature females. Most female in early maturity stages had sperm stored in the spermathecae, indicating that females copulate when still immature. There was no correlation between testis weight and the Needham complex, suggesting a protracted period of spermatophore production. Octopus insularis has a general gonadal development pattern similar to O. vulgaris, however, some differences were observed, as maturation at a smaller size, probably associated to a shorter life, and lower fecundity. The distinct reproductive features of O. insularis seem to be related to less variable conditions in the tropical environments. Management should take into account the differences and establish specific rules for the Northeast Brazil octopus fisheries.     

The structure and adhesive mechanism of octopus suckers

Octopus suckers consist of a tightly packed three-dimensionalarray of muscle with three major muscle fiber orientations:1) radial muscles that traverse the wall; 2) circular musclesarranged circumferentially around the sucker; and 3) meridionalmuscles oriented perpendicular to the circular and radial muscles.The sucker also includes inner and outer fibrous connectivetissue layers and an array of crossed connective tissue fibersembedded in the musculature. Adhesion results from reducingthe pressure inside the sucker cavity. This can be achievedby the three-dimensional array of muscle functioning as a muscular-hydrostat.Contraction of the radial muscles thins the wall, thereby increasingthe enclosed volume of the sucker. If the sucker is sealed toa surface the cohesiveness of water resists this expansion.Thus, the pressure of the enclosed water decreases instead.The meridional and circular muscles antagonize the radial muscles.The crossed connective tissue fibers may store elastic energy,providing an economical mechanism for maintaining attachmentfor extended periods. Measurements using miniature flush-mountedpressure transducers show that suckers can generate hydrostaticpressures below 0 kPa on wettable surfaces but cannot do soon non-wettable surfaces. Thus, cavitation, the failure of waterin tension, may limit the attachment force of suckers. As depthincreases, however, cavitation will cease to be limiting becauseambient pressure increases with depth while the cavitation thresholdis unchanged. Structural differences between suckers will thendetermine the attachment force.     

Bipedal locomotion in Octopus vulgaris: A complementary observation and some preliminary considerations

Lacking an external shell and a rigid endoskeleton, octopuses exhibit a remarkable flexibility in their movements. Bipedal locomotion is perhaps the most iconic example in this regard. Until recently, this peculiar mode of locomotion had been observed only in two species of tropical octopuses: Amphioctopus marginatus and Abdopus aculeatus. Yet, recent evidence indicates that bipedal walking is also part of the behavioral repertoire of the common octopus, Octopus vulgaris. Here we report a further observation of a defense behavior that encompasses both postural and locomotory elements of bipedal locomotion in this cephalopod. By highlighting differences and similarities with the other recently published report, we provide preliminary considerations with regard to bipedal locomotion in the common octopus.     

Receptor-mediated inhibition of octopamine-stimulated adenylate cyclase in the optic lobe of Octopus vulgaris

1. Inhibition of octopamine-stimulated adenylate cyclase was studied in the optic lobe of Octopus vulgaris.2. The octopamine antagonist, mianserin, and the dopamine D₂ agonists, PPHT and metergoline, induced dose-dependent inhibition of octopamine-stimulated adenylate cyclase activity.3. The binding of the tritiated benzazepine neuroleptic YM-09151-2 to octopus membranes and the displacement of [³H]YM-09151-2 by PPHT, metergoline and spiperone were consistent with the presence of a D₂-like dopamine receptor in the octopus optic lobe.4. The conclusion is drawn that octopamine-stimulated adenylate cyclase in the octopus is negatively regulated by a dopamine D₂-like receptor.     

Development of Biomimetic Squid-Inspired Suckers

Biomechanical properties of squid suckers were studied to provide inspiration for the development of sucker artefacts for a robotic octopus.Mechanical support of the rings found inside squid suckers was studied by bending tests.Tensile tests were carried out to study the maximum possible sucking force produced by squid suckers based on the strength of sucker stalks,normalized by the sucking areas.The squid suckers were also directly tested to obtain sucking forces by a special testing arrangement.Inspired by the squid suckers,three types of sucker artefacts were developed for the arm skin of an octopus inspired robot.The first sucker artefact made of knitted nylon sheet reinforced silicone rubber has the same shape as the squid suckers.Like real squid suckers,this type of artefact also has a stalk that is connected to the arm skin and a ring to give radial support.The second design is a straight cylindrical structure with uniform wall thickness made of silicone rubber.One end of the cylinder is directly connected to the arm skin and the other end is open.The final design of the sucker has a cylindrical base and a concave meniscus top.The meniscus was formed naturally using the surface tension of silicone gel,which leads to a higher level of the liquid around the edge of a container.The wall thickness decreases towards the tip of the sucker opening.Sucking forces of all three types of sucker artefacts were measured.Advantages and disadvantages of each sucker type were discussed.The final design of suckers has been implemented to the arm skin prototypes.     

Evidence for genetic differentiation of Octopus vulgaris (Mollusca, Cephalopoda) fishery populations from the southern coast of Brazil as revealed by microsatellites

Octopus vulgaris is a cephalopod species in several oceans and commonly caught by artisanal and industrial fisheries. In Brazil, O. vulgaris populations are mainly distributed along the southern coast and have been subjected to intensive fishing during recent years. Despite the importance of this marine resource, no genetic study has been carried out to examine genetic differences among populations along the coast of Brazil. In this study, 343 individuals collected by commercial vessels were genotyped at six microsatellite loci to investigate the genetic differences in O. vulgaris populations along the southern coast of Brazil. Genetic structure and levels of differentiation among sampling sites were estimated via a genotype assignment test and F-statistics. Our results indicate that the O. vulgaris stock consists of four genetic populations with an overall significant analogous F_ST (Φ_CT = 0.10710, P < 0.05) value. The genetic diversity was high with an observed heterozygosity of Ho = 0.987. The negative values of F_IS found for most of the loci examined suggested a possible bottleneck process. These findings are important for further steps toward more sustainable octopus fisheries, so that this marine resource can be preserved for long-term utilization.     

Immunolocalization of choline acetyltransferase of common type in the central brain mass of Octopus vulgaris

Acetylcholine, the first neurotransmitter to be identified in the vertebrate frog, is widely distributed among the animal kingdom. The presence of a large amount of acetylcholine in the nervous system of cephalopods is well known from several biochemical and physiological studies. However, little is known about the precise distribution of cholinergic structures due to a lack of a suitable histochemical technique for detecting acetylcholine. The most reliable method to visualize the cholinergic neurons is the immunohistochemical localization of the enzyme choline acetyltransferase, the synthetic enzyme of acetylcholine. Following our previous study on the distribution patterns of cholinergic neurons in the Octopus vulgaris visual system, using a novel antibody that recognizes choline acetyltransferase of the common type (cChAT), now we extend our investigation on the octopus central brain mass. When applied on sections of octopus central ganglia, immunoreactivity for cChAT was detected in cell bodies of all central brain mass lobes with the notable exception of the subfrontal and subvertical lobes. Positive varicosed nerves fibers where observed in the neuropil of all central brain mass lobes.Key words: invertebrate, cephalopod, choline acetyltransferase, neuron, immunohistochemistry.     

Contrasting macroscopic maturity staging with histological characteristics of the gonads in female Octopus vulgaris

We contrasted histological characteristics with a macroscopic maturity scale in Octopus vulgaris. Seven histological stages of maturation were identified, and a stereological method was used to develop a new histological maturity index (HMI). This index was related to the gonadosomatic index giving the possibility to estimate the histological stage of individual octopus without sampling the gonads. However, the existing macroscopic maturity scale produced some degree of overlap along the range of HMI, suggesting that this macroscale at this moment might be just good enough to separate immature from fully mature individuals. A histological maturity criterion based on the presence of a larger proportion of folding oocytes compared to earlier microstages resulted in a size at maturity of 1.5 kg. However, using two different macroscopic criteria, size at maturity was 1.3 and 2.3 kg. The estimate of size at maturity is therefore sensitive to the maturity criteria used. The maturation cycle of female O. vulgaris was seasonal, peaking in spring months and reaching a maximum of reproductive activity in April independently of the maturation criteria used. Oogenesis was reviewed and found to be an asynchronic process. Our results suggest that there is a need to examine all these issues in other cephalopod species.     

Methyl-end desaturases with ∆12 and ω3 regioselectivities enable the de novo PUFA biosynthesis in the cephalopod Octopus vulgaris

The interest in understanding the capacity of aquatic invertebrates to biosynthesise omega-3 (ω3) long-chain (≥C₂₀) polyunsaturated fatty acids (LC-PUFA) has increased in recent years. Using the common octopus Octopus vulgaris as a model species, we previously characterised a ∆5 desaturase and two elongases (i.e. Elovl2/5 and Elovl4) involved in the biosynthesis of LC-PUFA in molluscs. The aim of this study was to characterise both molecularly and functionally, two methyl-end (or ωx) desaturases that have been long regarded to be absent in most animals. O. vulgaris possess two ωx desaturase genes encoding enzymes with ∆12 and ω3 regioselectivities enabling the de novo biosynthesis of the C₁₈ PUFA 18:2ω6 (LA, linoleic acid) and 18:3ω3 (ALA, α-linolenic acid), generally regarded as dietary essential for animals. The O. vulgaris ∆12 desaturase (“ωx2”) mediates the conversion of 18:1ω9 (oleic acid) into LA, and subsequently, the ω3 desaturase (“ωx1”) catalyses the ∆15 desaturation from LA to ALA. Additionally, the O. vulgaris ω3 desaturase has ∆17 capacity towards a variety of C₂₀ ω6 PUFA that are converted to their ω3 PUFA products. Particularly relevant was the affinity of the ω3 desaturase towards 20:4ω6 (ARA, arachidonic acid) to produce 20:5ω3 (EPA, eicosapentaenoic acid), as supported by yeast heterologous expression, and enzymatic activity exhibited in vivo when paralarvae were incubated in the presence of [1-¹⁴C]20:4ω6. These results confirmed that several routes enabling EPA biosynthesis are operative in O. vulgaris whereas ARA and docosahexaenoic acid (DHA, 22:6ω3) should be considered essential fatty acids since endogenous production appears to be limited.     

Genetic structure of Octopus vulgaris (Cephalopoda,Octopodidae) in the central Mediterranean Sea inferred from the mitochondrial COIII gene

The polymorphism of the mitochondrial gene cytochrome oxidase III was studied in the Mediterranean octopus, Octopus vulgaris Cuvier, 1797. A total of 202 specimens from seven sampling sites were analysed with the aim of elucidating patterns of genetic structure in the central Mediterranean Sea and to give an insight into the phylogeny of the Octopus genus. Phylogenetic analyses showed that individuals from the central Mediterranean belong to the O. vulgaris species whose limits should nevertheless be clarified. Concerning genetic structure, two high-frequency haplotypes were present in all locations. The overall genetic divergence (Φ_ST = 0.05, P < 0.05) indicated a significant genetic structuring in the study area and an AMOVA highlighted a significant break between western and eastern Mediterranean basins (Φ_CT = 0.094, P < 0.05). Possible explanations for the observed patterns of genetic structuring are discussed with reference to their relevance for fisheries management.     

Morphological assessment of the Octopus vulgaris species complex evaluated in light of molecular‐based phylogenetic inferences

Cryptic species are common in the ocean, particularly among marine invertebrates such as octopuses. Delineating cryptic species is particularly problematic in octopus taxonomy where the plasticity recorded among taxonomic characters often results in low resolution at the species level. This study investigated the morphological relationships among seven phylogenetic clades (identified using cytochrome c oxidase subunit I) of the broadly distributed Octopus vulgaris species complex and close relatives. Morphological analyses in this study were successful in delimiting O. sinensis, Brazilian O. vulgaris and O. vulgaris sensu stricto, which was congruent with the molecular findings of this study. Analyses based on male morphology were successful in distinguishing 14 of 15 total pairwise comparisons and proved to be a more reliable indicator of species‐level relationships in comparison with female morphology. The majority of characters with the greatest discriminatory power were male sexual traits. Significant morphological differences were also recorded among sampling localities of conspecifics, with phenotype showing correlation with local environmental data. The findings of this study support the hypothesis that multiple O. vulgaris‐like species are currently being incorrectly treated under a single species name, O. vulgaris. Octopuses being exported globally under the name O. vulgaris are of extremely high fisheries market value and profile. Our findings have potentially significant implications for the naming and conservation of commercially harvested members of this species complex throughout their ranges.     

Home occupancy by male Octopus vulgaris in a large seawater tank

Pairs of individually recognizable male Octopus vulgaris were observed in a large seawater tank containing two suitable homes (brick pots or plastic buckets). None of the animals established exclusive occupancy of one home and for much of the time both animals were associated together at the same site. Usually one of the two homes was preferred and its occupant was most likely to be the larger animal, or the earlier resident if they were of equal size. Large animals were observed to take food forcefully from smaller octopus. An arm alignment interaction is described which, it is suggested, may be a means by which two octopuses establish their relative sizes.     

A dopamine- and octopamine-sensitive adenylate cyclase in the nervous system of Octopus vulgaris

• 1.1. Adenylate cyclase activity was assayed in the optic lobe of Octopus vulgaris.
• 2.2. Both octopamine and dopamine stimulate the octopus adenylate cyclase, apparently by competing with the same receptor site.
• 3.3. (±)-2-Amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene-HBr (6,7-ADTN) and a number of phenylethanolamine derivatives stimulate the octopus adenylate cyclase activity.
• 4.4. The dopamine D-1 antagonists R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine-HCl (SCH-23390) and (±)-7-bromo-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine-HCl (SKF-83566) are unable to antagonize the effects of dopamine and octopamine, and similarly ineffective is the agonist (±)-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine-7,8-diol-HCl (SKF-38393).
• 5.5. No detectable binding of labelled SCH-23390 occurs on membrane preparations from octopus optic lobe.

     

Developmental and physiological challenges of octopus (Octopus vulgaris) early life stages under ocean warming

The ability to understand and predict the effects of ocean warming (under realistic scenarios) on marine biota is of paramount importance, especially at the most vulnerable early life stages. Here we investigated the impact of predicted environmental warming (+3 °C) on the development, metabolism, heat shock response and antioxidant defense mechanisms of the early stages of the common octopus, Octopus vulgaris. As expected, warming shortened embryonic developmental time by 13 days, from 38 days at 18 °C to 25 days at 21 °C. Concomitantly, survival decreased significantly (~29.9 %). Size at hatching varied inversely with temperature, and the percentage of smaller premature paralarvae increased drastically, from 0 % at 18 °C to 17.8 % at 21 °C. The metabolic costs of the transition from an encapsulated embryo to a free planktonic form increased significantly with warming, and HSP70 concentrations and glutathione S-transferase activity levels were significantly magnified from late embryonic to paralarval stages. Yet, despite the presence of effective antioxidant defense mechanisms, ocean warming led to an augmentation of malondialdehyde levels (an indicative of enhanced ROS action), a process considered to be one of the most frequent cellular injury mechanisms. Thus, the present study provides clues about how the magnitude and rate of ocean warming will challenge the buffering capacities of octopus embryos and hatchlings’ physiology. The prediction and understanding of the biochemical and physiological responses to warmer temperatures (under realistic scenarios) is crucial for the management of highly commercial and ecologically important species, such as O. vulgaris.     

Co(II)-substituted Octopus vulgaris hemocyanin

Co(II)-substituted hemocyanin (Co(II)Hc) of the octopus, Octopus vulgaris, has been prepared by dialysis of apohemocyanin against Co(II·) ion and subsequent Chelex-treatment. The blue 50%-Co(II)Hc (half-apo Co(II)Hc), in which binuclear coppers are replaced in the hemocyanin by a single Co(II), exhibits two absorption maxima at 560 (?_Co=250) and 594 nm (?_Co=320 M^?1 cm^?1) and a shoulder near 610 nm, all of which are attributed to a dd transition of high spin Co(II) (S=3/2) with a tetrahedral geometry. The magnetic circular dichroism (MCD) spectrum in this region also suggests the existence of a tetrahedral Co(II) species in the protein. The visible absorption and MCD spectra of octopus 50%-Co(II)Hc are quite similar to those of squid 50%-Co(II)Hc described in the previous paper (S. Suzuki, J. Kino, M. Kimura, W. Mori and A. Nakahara, Inorg. Chim. Acta, 66, 41 (1982)). The formation of half-apo Co(II)Hc demonstrates that the binuclear copper sites in native octopus hemocyanin may differ from each other in coordination geometry, as in other molluscan hemocyanins, squid and snail hemocyanins. The coordination environment of the active-site Co(II) substituted for Cu in the octopus hemocyanin is the same as that of the corresponding active site of the squid hemocyanin.     

Ventilation rate and arousal in the Octopus

Observations on undisturbed, individually isolated, Octopus vulgaris Cuvier show that ventilation rate is strongly related to “arousal” or activation. Ventilation rate may be recorded visually by a screened observer or remotely, using carbon electrodes connected to a pen recorder via an impedance coupler to register the associated water movements in a small experimental chamber. The method allows a quantifiable behavioural characteristic to be measured objectively. It has been used to demonstrate that O. vulgaris responds to certain chemical stimuli in flowing sea water with a sharply increased ventilation rate. This result is taken to show distance chemoreception in this cephalopod.