An experimental method for evoking and characterizing dynamic color patterning of cuttlefish during prey capture

© The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Kim, D., Buresch, K. C., Hanlon, R. T., & Kampff, A. R. An experimental method for evoking and characterizing dynamic color patterning of cuttlefish during prey capture. Journal of Biological Methods, 9(2), (2022): e161, https://doi.org/10.14440/jbm.2022.386.Cuttlefish are active carnivores that possess a wide repertoire of body patterns that can be changed within milliseconds for many types of camouflage and communication. The forms and functions of many body patterns are well known from ethological studies in the field and laboratory. Yet one aspect has not been reported in detail: the category of rapid, brief and high-contrast changes in body coloration (“Tentacle Shot Patterns” or TSPs) that always occur with the ejection of two ballistic tentacles to strike live moving prey (“Tentacles Go Ballistic” or TGB moment). We designed and tested a mechanical device that presented prey in a controlled manner, taking advantage of a key stimulus for feeding: motion of the prey. High-speed video recordings show a rapid transition into TSPs starting 114 ms before TGB (N = 114). TSPs are then suppressed as early as 470–500 ms after TGB (P < 0.05) in unsuccessful hunts, while persisting for at least 3 s after TGB in successful hunts. A granularity analysis revealed significant differences in the large-scale high-contrast body patterning present in TSPs compared to the camouflage body pattern deployed beforehand. TSPs best fit the category of secondary defense called deimatic displaying, meant to briefly startle predators and interrupt their attack sequence while cuttlefish are distracted by striking prey. We characterize TSPs as a pattern category for which the main distinguishing feature is a high-contrast signaling pattern with aspects of Acute Conflict Mottle or Acute Disruptive Pattern. The data and methodology presented here open opportunities for quantifying the rapid neural responses in this visual sensorimotor set of behaviors.KCB and RTH acknowledge partial support from the Sholley Foundation

Octopus arms exhibit exceptional flexibility

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Kennedy, E. B. L., Buresch, K. C., Boinapally, P., & Hanlon, R. T. Octopus arms exhibit exceptional flexibility. Scientific Reports, 10(1), (2020): 20872. doi:10.1038/s41598-020-77873-7.The octopus arm is often referred to as one of the most flexible limbs in nature, yet this assumption requires detailed inspection given that this has not been measured comprehensively for all portions of each arm. We investigated the diversity of arm deformations in Octopus bimaculoides with a frame-by-frame observational analysis of laboratory video footage in which animals were challenged with different tasks. Diverse movements in these hydrostatic arms are produced by some combination of four basic deformations: bending (orally, aborally; inward, outward), torsion (clockwise, counter-clockwise), elongation, and shortening. More than 16,500 arm deformations were observed in 120 min of video. Results showed that all eight arms were capable of all four types of deformation along their lengths and in all directions. Arms function primarily to bring the sucker-lined oral surface in contact with target surfaces. Bending was the most common deformation observed, although the proximal third of the arms performed relatively less bending and more shortening and elongation as compared with other arm regions. These findings demonstrate the exceptional flexibility of the octopus arm and provide a basis for investigating motor control of the entire arm, which may aid the future development of soft robotics.We gratefully acknowledge funding from Grant N00014-19-1-2445 from the Office of Naval Research, Tom McKenna and Marc Steinberg, Program Managers. We also thank the staff of the Marine Resources Center at MBL for assistance with water quality measurements, seawater system maintenance, and collection of food items for octopuses

Expression of squid iridescence depends on environmental luminance and peripheral ganglion control

Author Posting. © The Author(s), 2013. This is the author's version of the work. It is posted here by permission of Company of Biologists for personal use, not for redistribution. The definitive version was published in Journal of Experimental Biology 217 (2014):850-858, doi:10.1242/​jeb.091884.Squids display impressive changes in body coloration that are afforded by two types of dynamic skin elements: structural iridophores (which produce iridescence) and pigmented chromatophores. Both color elements are neurally controlled, but nothing is known about the iridescence circuit, or the environmental cues, that elicit iridescence expression. To tackle this knowledge gap, we performed denervation, electrical stimulation and behavioral experiments using the long-fin squid, Doryteuthis pealeii. We show that while the pigmentary and iridescence circuits originate in the brain, they are wired differently in the periphery: (i) the iridescence signals are routed through a peripheral center called the stellate ganglion and (ii) the iridescence motorneurons likely originate within this ganglion (as revealed by nerve fluorescence dye fills). Cutting the inputs to the stellate ganglion that descend from the brain shifts highly reflective iridophores into a transparent state. Taken together, these findings suggest that although brain commands are necessary for expression of iridescence, integration with peripheral information in the stellate ganglion could modulate the final output. We also demonstrate that squids change their iridescence brightness in response to environmental luminance; such changes are robust but slow (minutes to hours). The squid's ability to alter its iridescence levels may improve camouflage under different lighting intensities.This research was supported by the ONR Basic Research Challenge grant no. N00014-10-1-0989 and by the AFOSR grant FA9950090346.2015-03-1

How visual edge features influence cuttlefish camouflage patterning

AbstractRapid adaptive camouflage is the primary defense of soft-bodied cuttlefish. Previous studies have shown that cuttlefish body patterns are strongly influenced by visual edges in the substrate. The aim of the present study was to examine how cuttlefish body patterning is differentially controlled by various aspects of edges, including contrast polarity, contrast strength, and the presence or absence of “line terminators” introduced into a pattern when continuous edges are fragmented. Spatially high- and low-pass filtered white or black disks, as well as isolated, continuous and fragmented edges varying in contrast, were used to assess activation of cuttlefish skin components. Although disks of both contrast polarities evoked relatively weak disruptive body patterns, black disks activated different skin components than white disks, and high-frequency information alone sufficed to drive the responses to white disks whereas high- and low-frequency information were both required to drive responses to black disks. Strikingly, high-contrast edge fragments evoked substantially stronger body pattern responses than low-contrast edge fragments, whereas the body pattern responses evoked by high-contrast continuous edges were no stronger than those produced by low-contrast edges. This suggests that line terminators vs. continuous edges influence expression of disruptive body pattern components via different mechanisms that are controlled by contrast in different ways

Tactical decisions for changeable cuttlefish camouflage : visual cues for choosing masquerade are relevant from a greater distance than visual cues used for background matching

Author Posting. © Marine Biological Laboratory, 2015. This article is posted here by permission of Marine Biological Laboratory for personal use, not for redistribution. The definitive version was published in Biological Bulletin 229 (2015): 160-166.Cuttlefish use multiple camouflage tactics to evade their predators. Two common tactics are background matching (resembling the background to hinder detection) and masquerade (resembling an uninteresting or inanimate object to impede detection or recognition). We investigated how the distance and orientation of visual stimuli affected the choice of these two camouflage tactics. In the current experiments, cuttlefish were presented with three visual cues: 2D horizontal floor, 2D vertical wall, and 3D object. Each was placed at several distances: directly beneath (in a circle whose diameter was one body length (BL); at zero BL [(0BL); i.e., directly beside, but not beneath the cuttlefish]; at 1BL; and at 2BL. Cuttlefish continued to respond to 3D visual cues from a greater distance than to a horizontal or vertical stimulus. It appears that background matching is chosen when visual cues are relevant only in the immediate benthic surroundings. However, for masquerade, objects located multiple body lengths away remained relevant for choice of camouflage.This work was funded by DARPA/DSO grant no. W15P7T-13-D-CT04

Vertical visual features have a strong influence on cuttlefish camouflage

Author Posting. © Marine Biological Laboratory, 2013. This article is posted here by permission of Marine Biological Laboratory for personal use, not for redistribution. The definitive version was published in Biological Bulletin 224 (2013): 110-118.Cuttlefish and other cephalopods use visual cues from their surroundings to adaptively change their body pattern for camouflage. Numerous previous experiments have demonstrated the influence of two-dimensional (2D) substrates (e.g., sand and gravel habitats) on camouflage, yet many marine habitats have varied three-dimensional (3D) structures among which cuttlefish camouflage from predators, including benthic predators that view cuttlefish horizontally against such 3D backgrounds. We conducted laboratory experiments, using Sepia officinalis, to test the relative influence of horizontal versus vertical visual cues on cuttlefish camouflage: 2D patterns on benthic substrates were tested versus 2D wall patterns and 3D objects with patterns. Specifically, we investigated the influence of (i) quantity and (ii) placement of high-contrast elements on a 3D object or a 2D wall, as well as (iii) the diameter and (iv) number of 3D objects with high-contrast elements on cuttlefish body pattern expression. Additionally, we tested the influence of high-contrast visual stimuli covering the entire 2D benthic substrate versus the entire 2D wall. In all experiments, visual cues presented in the vertical plane evoked the strongest body pattern response in cuttlefish. These experiments support field observations that, in some marine habitats, cuttlefish will respond to vertically oriented background features even when the preponderance of visual information in their field of view seems to be from the 2D surrounding substrate. Such choices highlight the selective decision-making that occurs in cephalopods with their adaptive camouflage capability.This work was funded by the United States Department of Defense (grant number W911- NF-07-D-0001)

Experimental evidence that ovary and oviducal gland extracts influence male agonistic behavior in squids

Author Posting. © Marine Biological Laboratory, 2004. This article is posted here by permission of Marine Biological Laboratory for personal use, not for redistribution. The definitive version was published in Biological Bulletin 206 (2004): 1-3.Recent investigations of sensory and behavioral cues that initiate sexual selection processes in the squid Loligo pealeii have determined that egg capsules deposited on the substrate provide a strong visual and chemotactile stimulus to males, even in the absence of females (1, 2, 3). The visual stimulus of egg capsules attracts males to the eggs, and when the males touch the eggs, they encounter a chemical stimulus that leads to highly aggressive fighting behavior. We have recently demonstrated that egg capsule extracts implanted in artificial egg capsules elicit this aggressive behavior (4). In this communication, we present evidence that the salient chemical factor originates in the ovary and perhaps the oviducal gland of the female reproductive tract

Cuttlefish camouflage: The effects of substrate contrast and size in evoking uniform, mottle or disruptive body patterns

AbstractCuttlefish are cephalopod molluscs that achieve dynamic camouflage by rapidly extracting visual information from the background and neurally implementing an appropriate skin (or body) pattern. We investigated how cuttlefish body patterning responses are influenced by contrast and spatial scale by varying the contrast and the size of checkerboard backgrounds. We found that: (1) at high contrast levels, cuttlefish body patterning depended on check size; (2) for low contrast levels, body patterning was independent of “check” size; and (3) on the same check size, cuttlefish fine-tuned the contrast and fine structure of their body patterns, in response to small contrast changes in the background. Furthermore, we developed an objective, automated method of assessing cuttlefish camouflage patterns that quantitatively differentiated the three body patterns of uniform/stipple, mottle and disruptive. This study draws attention to the key roles played by background contrast and particle size in determining an effective camouflage pattern

Defensive responses of cuttlefish to different teleost predators

Author Posting. © Marine Biological Laboratory, 2013. This article is posted here by permission of Marine Biological Laboratory for personal use, not for redistribution. The definitive version was published in Biological Bulletin 225 (2013): 161-174.We evaluated cuttlefish (Sepia officinalis) responses to three teleost predators: bluefish (Pomatomus saltatrix), summer flounder (Paralichthys dentatus), and black seabass (Centropristis striata). We hypothesized that the distinct body shapes, swimming behaviors, and predation tactics exhibited by the three fishes would elicit markedly different antipredator responses by cuttlefish. Over the course of 25 predator-prey behavioral trials, 3 primary and 15 secondary defense behaviors of cuttlefish were shown to predators. In contrast, secondary defenses were not shown during control trials in which predators were absent. With seabass—a benthic, sit-and-pursue predator—cuttlefish used flight and spent more time swimming in the water column than with other predators. With bluefish—an active, pelagic searching predator—cuttlefish remained closely associated with the substrate and relied more on cryptic behaviors. Startle (deimatic) displays were the most frequent secondary defense shown to seabass and bluefish, particularly the Dark eye ring and Deimatic spot displays. We were unable to evaluate secondary defenses by cuttlefish to flounder—a lie-and-wait predator—because flounder did not pursue cuttlefish or make attacks. Nonetheless, cuttlefish used primary defense during flounder trials, alternating between cryptic still and moving behaviors. Overall, our results suggest that cuttlefish may vary their behavior in the presence of different teleost predators: cryptic behaviors may be more important in the presence of active searching predators (e.g., bluefish), while conspicuous movements such as swimming in the water column and startle displays may be more prevalent with relatively sedentary, bottom-associated predators (e.g., seabass).This project was funded by a United States Department of Defense, Defense Advanced Research Projects Agency, Defense Sciences Office (DARPA DSO) Grant (HR0011-09- 1-0017)

Multiple genetic stocks of longfin squid Loligo pealeii in the NW Atlantic : stocks segregate inshore in summer, but aggregate offshore in winter

Author Posting. © Inter-Research, 2006. This article is posted here by permission of Inter-Research for personal use, not for redistribution. The definitive version was published in Marine Ecology Progress Series 310 (2006): 263-270, doi:10.3354/meps310263.The longfin squid Loligo pealeii is distributed widely in the NW Atlantic and is the target of a major fishery. A previous electrophoretic study of L. pealeii was unable to prove genetic differentiation, and the fishery has been managed as a single unit stock. We tested for population structure using 5 microsatellite loci. In early summer (June), when the squids had migrated inshore to spawn, we distinguished 4 genetically distinct stocks between Delaware and Cape Cod (ca. 490 km); a 5th genetic stock occurred in Nova Scotia and a 6th in the northern Gulf of Mexico. One of the summer inshore stocks did not show genetic differentiation from 2 of the winter offshore populations. We suggest that squids from summer locations overwinter in offshore canyons and that winter offshore fishing may affect multiple stocks of the inshore fishery. In spring, squids may segregate by genetic stock as they undertake their inshore migration, indicating an underlying mechanism of subpopulation recognition.We acknowledge funding from WHOI Sea Grant NA16RG2273, the Massachusetts Environmental Trust (#98-04), and the Sholley Foundation