Depth cues, behavioural context, and natural illumination: some potential limitations of video playback techniques

By expanding on issues raised by D’Eath (1998), I address in this article three aspects of vision that are difficult to reproduce in the video- and computer-generated images used in experiments, in which images of conspecifics or of predators are replayed to animals. The lack of depth cues derived from binocular stereopsis, from accommodation, and from motion parallax may be one of the reasons why animals do not respond to video displays in the same way as they do to real conspecifics or to predators. Part of the problem is the difficulty of reproducing the closed-loop nature of natural vision in video playback experiments. Every movement an animal makes has consequences for the pattern of stimulation on its retina and this ”optic flow” in turn carries information about both the animal’s own movement and about the three-dimensional structure of the environment. A further critical issue is the behavioural context that often determines what animals attend to but that may be difficult to induce or reproduce in an experimental setting. I illustrate this point by describing some visual behaviours in fiddler crabs, in which social and spatial context define which part of the visual field a crab attends to and which visual information is used to guide behaviour. I finally mention some aspects of natural illumination that may influence how animals perceive an object or a scene: shadows, specular reflections, and polarisation reflections. Received: 23 November 1999 / Received in revised form: 9 February 2000 / Accepted: 10 February 2000     

An evaluation of video playback using Xiphophorus helleri

Video playback is being increasingly used as a technique for behavioural research. The importance of critically evaluating the effectiveness of video playback is clear, as available video technology is not designed for nonhuman visual systems. We discuss several aspects concerning the perception of video images that could lead to inconclusive or erroneous results. Researchers should verify that behaviour observed in response to video playback is comparable to behaviour observed in response to live animals. We conducted such a verification using live and video playback methods to measure female response to swords of varying lengths in the green swordtail, Xiphophorus helleri. Using both methods, female response appeared to be an increasing function of male sword length. Females did not differ in their response to live and video versions of noncourting, noninteractive males, however, females tended to prefer video playbacks of males with longer swords, a result that has also been found in experiments using live males. These results suggest that females express the same qualitative mating preference, but not necessarily the same quantitative preference, for sword length when viewing video stimuli. Several methodological factors that may contribute to an apparent difference in the strength of the preference are discussed. Despite these differences, both methods produced comparable results; female response to sworded males tended to increase as sword length increased. These experiments demonstrate that video playback is an effective method to measure female preferences accurately in X. helleri and provide an example of how video playback can be evaluated in other species. Copyright 2000 The Association for the Study of Animal Behaviour.     

Some comments on visual perception and the use of video playback in animal behavior studies

Video playback experiments are potentially powerful tools in behavioral research. A video screen mimics natural color, brightness, texture, and motion to humans (for which it was designed) because monitors stimulate human photoreceptors in approximately the same relative proportions as the stimuli that they mimic. Because most animals have vision that is very different from that of humans their cones may be stimulated very differently from ours, and an image that looks excellent to us may be unrecognizable to them, and vice versa. In this article we summarize how the simulation of a monitor works and the ways it can go wrong, using a bird and a fish model retina as examples. Finally we make some recommendations for minimizing some of these problems. Received: 10 November 1999 / Received in revised form: 22 February 2000 / Accepted: 2 March 2000     

Are there lessons from negative results in studies using video playback?

The use of video playback is a growing practice in animal behaviour. Studies that lead to unpredicted negative results are often considered failures and remain unpublished. In this article I review some of the published and unpublished studies that have led to negative results. Analysis of these studies suggests that no particular problem is responsible for more than a small number of negative results. I suggest further investigation of the potential causes of these negative results to assess their importance to the outcome of video playback experiments. Received: 31 January 2000 / Received in revised form: 27 March 2000 / Accepted: 28 March 2000     

Playback experiments: design and analysis

The scientific value of the outcome of an experiment is closely related to its design and analysis. This article deals with the design issues of pseudoreplication (whether the experimental design has the statistical features needed to answer the question as posed) and execution errors (problems arising from how the experiment was conducted). Three issues of analysis are also dealt with: the number and type of response measures to record; how measures should, and should not, be combined into a single response measure; and how to interpret an apparent lack of response. Interactive playback is considered separately because it raises its own specific design and analysis issues. Although the examples generally refer to video playback, these issues are common to all experiments in behaviour. Received: 23 September 1999 / Received in revised form: 24 February 2000 / Accepted: 25 February 2000     

Computer-manipulated stimuli as a research tool in Mozambique tilapia <Emphasis Type="Italic">Oreochromis mossambicus</Emphasis>

Multimodal communication is essential in social interactions in cichlid fish, including conspecifics’ recognition, agonistic interactions and courtship behaviour. Computer-manipulated image stimuli and sound playback offer powerful tools to assess the relative relevance of visual and acoustic stimuli in fish behavioural studies, but these techniques require validation for each taxon. The aim of the present study was to investigate whether Mozambique tilapia Oreochromis mossambicus responds to computer-manipulated visual stimuli and acoustic playback. Six experiments were conducted: computer animation playback, video playback, interaction with a mirror, presentation of a live male in a jar alone and combined with courting sound playback or with white noise playback. Individual agonistic interactions (lateral displays, up and down swimming, butting) and courting behaviours (tilting leading, digging) were tallied for each experiment. Our results suggest that non-interactive computer-manipulated visual stimuli is not a suitable tool in behavioural research with Mozambique tilapia. In contrast, interaction with a live male in a jar seems to remain the best visual research instrument inducing significant strong behavioural responses. Although none or only a few agonistic interactions were observed towards video playbacks or computer animations, such interactions significantly increased towards a male in jar and were modulated by courtship sound playback, suggesting the additional relevance of sound playback as a tool in behavioural research with Mozambique tilapia, including the study of multimodal signalling.     

Dichromatic colour vision in an Australian marsupial, the tammar wallaby

Despite earlier assertions that most mammals are colour blind, colour vision has in recent years been demonstrated in a variety of eutherian mammals from a wide range of different orders. This paper presents the first behavioural evidence from colour discrimination experiments, that an Australian marsupial, the tammar wallaby (Macropus eugenii), has dichromatic colour vision. In addition, the experiments show that the wallabies readily learn the relationship between the presented colours rather than the absolute hues. This provides a sensitive method to measure the location of the neutral-point, which is the wavelength of monochromatic light that is indistinguishable from white. This point is a diagnostic feature for dichromats. The spectral sensitivity of the wallabies' middle-wavelength-sensitive photoreceptor is known (peak: 539 nm) and the behavioural results imply that the sensitivity of the short-wavelength-sensitive receptor must be near 420 nm. These spectral sensitivities are similar to those found in eutherian mammals, supporting the view that the earliest mammals had dichromatic colour vision. Accepted: 18 July 1999     

Sex recognition in zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>)

We investigated sex recognition in female zebrafish (Danio rerio) to better understand the underlying sensory mechanisms and identify male secondary sexual traits. Females were simultaneously presented with two fish, a male and a female, in a flow-chamber apparatus, and females’ relative attraction towards males was observed under different conditions. With domesticated fish, females were more attracted to males when presented with both visual and chemosensory cues from stimulus fish. They still discriminated the sexes when only visual cues were provided, but not when white ambient light was changed to yellow, indicating that colour plays a role. Sex discrimination under yellow light was improved when chemosensory cues were also provided. Surprisingly, females’ attraction to males was not more pronounced in the morning when mating occurs. Domesticated females discriminated the sexes when presented with wild-derived, as well as domesticated fish, whereas wild-derived females did not show any biases for domesticated or wild-derived males. Behavioural observations indicated that the wild-derived females were distressed, which explains their lack of attraction to males. In summary, domesticated female zebrafish discriminated the sexes using both visual (body colour) and olfactory cues; however, wild-derived zebrafish were too distressed for behavioural experiments under these laboratory conditions.     

Corn bunting (<Emphasis Type="Italic">Miliaria calandra</Emphasis>) males respond differently to alternating and overlapping playback of song

Interactive playback experiments were used to study the signal value to the corn bunting, Miliaria calandra, of alternating and overlapping singing. We subjected 15 males to two stimuli that differed in the temporal pattern of song playback (alternating or overlapping). We measured eight characteristics of the males’ response in two categories—song output and movements. Overlapping and alternating playback elicited a similar song response, characteristic of highly aroused males. Song response correlated positively with males’ singing activity before playback, irrespective of stimulus. There were significant differences between latency of approach to the loudspeaker and number of flights. Birds approached the loudspeaker more quickly and spent more time close to it when playback alternated with their songs. The results suggest overlapping song could be interpreted as a stronger threat but elicits a more cautious, rather than stronger, response than the alternating pattern. Males were found to shorten songs during the playback compared with songs sung before and after stimulation. The only predictor of degree of song shortening was song activity before the playback began. It should, therefore, be regarded as a signal which is related to escalated, close-distance counter-singing.     

Design considerations and techniques for constructing video stimuli

Techniques for constructing video playback stimuli fall into five categories. The first three involve manipulating video sequences: (1) edited video is a temporal rearrangement of raw footage, (2) processed video applies global filtering algorithms to edited video, and (3) frame-manipulated video involves manually altering individual frames. The last two, (4) exemplar-based animation and (5) parameter-based animation, are synthetic models derived from visual parameters based on a single exemplar and sample data, respectively. Image-based approaches are straightforward to apply and preserve fine spatiotemporal detail. Synthetic stimuli are desirable when a large number of manipulations are called for and to ensure individual stimuli reflect population characteristics. Received: 13 December 1999 / Received in revised form: 25 February 2000 / Accepted: 1 March 2000     

Responses of golden-backed uakaris, <Emphasis Type="Italic">Cacajao melanocephalus</Emphasis>, to call playback: implications for surveys in the flooded Igapó forest

We investigated the potential use of call playback for surveying the elusive golden-backed uakari (Cacajao melanocephalus), a diurnal social primate that typically inhabits the relatively inaccessible black-water swamp forests of the upper Amazon (Igapó forest) during the wet season. Furthermore, our objective was to understand better the functions of vocalisations of these rare and threatened monkeys. We focussed on the “tchó” call, a loud vocalisation that varies in temporal and frequency characteristics according to behavioural context and individual signaller (Bezerra et al. in Int J Primatol, 2010b). We investigated the vocal and behavioural responses of golden-backed uakaris from three Igapó areas to three different playback stimuli: own group calls (OGC); neighbouring group calls (NGC); and control (background noise). The call stimuli were used to simulate the unexpected presence of monkeys from the monkeys’ own and neighbouring Igapó areas. Playback of OGC and NGC increased calling and altered behaviour in golden-backed uakaris, whereas no noticeable response occurred to the control stimuli. Furthermore, there was no evidence of habituation to the call stimuli and the natural behavioural pattern of the groups was not obviously affected. Hence, call playback seems to be a valuable tool for locating these elusive monkeys in river surveys. Most of the agonistic-related behaviour observed after NGC playback was also observed after OGC playback. As golden-backed uakaris are not territorial, the presence of strange conspecifics to the area may not necessarily represent a more threatening situation. Our methods and results may also be useful for surveys of other primate species living in the Igapó forest and in other habitat types.     

Digital video playback and visual communication in lizards

Experimental analyses of dynamic visual signals have to overcome the technical obstacle of reproducing complex motor patterns such as those found in courtship and threat displays. Video playback offers a potential solution to this problem, but it has recently been criticized because of sensory differences between humans and nonhuman animals, which suggest that video stimuli might be perceived as deficient relative to live conspecifics. Quantitative comparisons are therefore necessary to determine whether video sequences reliably evoke natural responses. Male Jacky dragons, Amphibolurus muricatus, compete for territories using complex displays delivered in a rapid stereotyped sequence. We evaluated video playback as a technique for studying this visual signal. Digital video sequences depicting a life-sized displaying male were indistinguishable from live male conspecifics in the rate and structure of aggressive displays evoked. Other measures of social behaviour suggested that video stimuli were more effective in this context. Lizards produced significantly more appeasement displays and had higher rates of substrate licking and locomotor activity in response to video playback than to confined male opponents, which failed to produce aggressive displays. Lizards tracked temporal changes in the display rate of video stimuli and were also sensitive to individual differences in morphology and behaviour between video exemplars. These results show that video stimuli are appropriate for the experimental analysis of Jacky dragon aggressive displays. We compare the potential shortcomings of video playback with those of other techniques and conclude that no approach offers a panacea, but that several have complementary characteristics. Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.     

Interactive video playback and opponent assessment in lizards

Video playback has been used to explore many issues in animal communication, but the scope of this work has been constrained by the lack of stimulus-subject interaction. In many natural contexts, each participant's signalling behaviour is dependent from moment-to-moment on that of the other. Analyses of acoustic communication demonstrate the value of reproducing such social contingencies. We assessed the utility of interactive playback for studies of visual signalling by comparing the responses of male Jacky dragons, Amphibolurus muricatus, to interactive and non-interactive digital video playbacks of a life-sized conspecific. Displays produced by lizards in the interactive condition had the effect of suppressing the aggressive display of their simulated opponent. Each stimulus sequence generated during an interactive playback was subsequently played to a size-matched control animal. Males that could interact with the video stimulus responded principally with aggressive displays, while those that could not produced a mixture of aggressive and appeasement signals. Adding a degree of receiver responsiveness is hence sufficient to alter the type of signal evoked, even when video stimuli are physically identical. Interactive playback permits the experimental study of a broader range of theoretical topics and can enhance the realism of video stimuli.     

The significance of sound interception to males of the bicolor damselfish,Pomacentrus partitus,during courtship

Synopsis Playback experiments conducted in the field and the laboratory demonstrated the use of sound interception in bicolor damselfish,Pomacentrus partitus. Two courtship sounds produced by male bicolors: the ‘chirp’ which occurs at the initiation of courting and the ‘grunt’ which occurs near the termination, just prior to spawning, were found to hold different meanings to intercepting male competitors. Males responded to grunt playback with directional swimming towards the sound source and increased courtship behavior. No directional response was observed during chirp playback. The grunt sound appears to indicate that a spawn-ready female is present near the sound source. Males therefore move towards it, likely to interfere with the imminent spawning or to gain a possible spawning partner. Such is not observed in response to chirps, as little advantage would be gained by moving to the source of a sound which is known to serve as an ‘advertisement’ or ‘territorial keep-out’ signal. Thus, upon intercepting a neighbor's chirps, males exhibit courting within their own territory, or directly court a nearby female.     

Luminance-dependence of spatial vision in budgerigars (<Emphasis Type="Italic">Melopsittacus undulatus</Emphasis>) and Bourke’s parrots (<Emphasis Type="Italic">Neopsephotus bourkii</Emphasis>)

Budgerigars (Melopsittacus undulatus) and Bourke’s parrots (Neopsephotus bourkii) are closely related birds with different activity patterns. Budgerigars are strictly diurnal while Bourke’s parrots are active in dim twilight. Earlier studies show that the intensity threshold of colour vision is similar in both species while Bourke’s parrots have larger eyes with a higher density of rods than budgerigars. In this study, we investigate whether this could be an adaptation for better spatial vision in dim light. We used two alternative forced-choice experiments to determine the spatial acuity of both species at light intensities ranging from 0.08 to 73 cd/m². We also determined the spatial contrast sensitivity function (CSF) for bright light in Bourke’s parrots and compare it to existing data for budgerigars. The spatial acuity of Bourke’s parrots was found to be similar to that of budgerigars at all light levels. Also the CSF of Bourke’s parrots is similar to that of budgerigars with a sensitivity peak located between 2.1 and 2.6 cycles/degree. Our findings do not support the hypothesis that Bourke’s parrots have superior spatial acuity in dim light compared to budgerigars and the adaptive value of the relatively rod-rich and large eyes of Bourke’s parrots remains unclear.     

Male or female soldiers? An evaluation of several factors which may influence soldier sex ratio in lower termites

In termites, the soldiers’ sex ratio is often biased toward one sex. Unlike in the Hymenoptera, this bias cannot easily be explained by relatedness asymmetries because termites are diploid. Matsuura proposed that when large body size is adaptive for colony defence (e.g. in case of phragmotic defence) then the larger sex (given sexual size dimorphism exists) should be more likely to reach a threshold size and develop into soldiers. This would explain biased sex ratios. Matsuura validated his hypothesis for four Reticulitermes species. Here, we tested his hypothesis for two species of Cryptotermes with phragmotic defence. These drywood termites have a life type that is thought to be ancestral in termite’s evolution, thus giving us potential insights into the evolution of the soldier caste. In one of these species, the sex ratio of soldiers was highly female biased, but we could not support Matsuura’s hypothesis. Both species lacked sexual size dimorphism in all castes. Additionally, in both species, the sex ratio of helpers and sexuals did not deviate from a 1:1 ratio, and hence can also not account for the bias observed in soldiers. However, this study showed that there were behavioural differences between the sexes in both species, which could shed some light on biased sex ratio in soldiers. Our findings also indicate that the developmental pathway taken by individuals reflects a ‘decision’ at the colony level. The discovery of behavioural differences between sexes in termites should open the way to similar studies in other taxa with helpers/ workers of both sexes, as it might reveal more task partitioning in colonies than previously thought and it raises questions concerning the selective pressures that acted on caste evolution in termites. Received 30 October 2007; revised 17 January and 27 February; accepted 4 March 2008.     

The turn-back-and-look behaviour: bee versus robot

 Honeybees and social wasps departing from a novel food source perform stereotype flight manoeuvres, termed the turn-back-and-look behaviour (TBL). Based on results of behavioural studies, it is proposed that the image motion generated by the TBL provides the insect with information about the three-dimensional structure of the goal's surroundings, thus enabling it to select reliable landmarks that will guide it to the goal upon return. The colour, shape, and size of landmarks, on the other hand, are learned mainly during arrival at the food source. However, when bees are prevented from learning these cues on arrival, they learn them during the TBL, despite the fact that this performance does not require the use of image motion. A recently developed model shows that landmark learning can indeed be accomplished during the TBL by exploiting cues others than image motion. A mobile robot equipped with the appropriate software selects, during the TBL, reliable marks and returns to the site of departure from different locations by accomplishing image matching along a two-dimensional vector field. Received: 7 May 1999 / Accepted in revised form: 20 March 2000     

Cryopreservation of Panax ginseng cells

The impacts of cryoprotectants (CP) and cell status during the growth cycle on Panax ginseng cell viability during cryopreservation were investigated. The ginseng cells used had a 5–7 times proliferation rate (compared with inoculum) in 2–3 weeks and were subcultured at 2- and 4-week intervals in liquid and on solid media, respectively. After testing various CP solutions of glycerol, dimethylsulphoxide, ethylene glycol and sucrose, a combination of 10% (v/v) glycerol and 4% (w/v) sucrose was selected for its least cytotoxicity and highest cell viability after thawing. With this CP solution, cells throughout the growth cycle exhibited a ‘U’-shaped fluctuation of post-thaw cell viability. The highest viability (86.5%) occurred during the lag phase from cells already maintained in suspension culture and then in the late exponential phase (61.7%); the lowest level of 15.4% was in the mid-exponential phase. Callus freshly transferred to liquid medium showed a less obvious fluctuation pattern. The recovered cells were brown-to-reddish at first and gradually returned to a light yellow colour after several subcultures. Received: 1 October 1998 / Revision received: 6 January 2000 / Accepted: 11 January 2000     

Thermoregulatory differences between phenotypes in the speckled wood butterfly: hot perchers and cold patrollers?

Males of the speckled wood butterfly Pararge aegeria L. (Satyrinae), actively search for females (“patrolling”) or wait for them at particular places (“perching”). Darker males are more likely to patrol than pale ones, which are mainly territorial perchers. We studied whether this morphological variation relates to thermoregulatory differences. The relationship between thoracic temperature and ambient temperature differed between the colour types under natural conditions: darker males had on average lower body temperatures than paler males. Different activities (e.g. resting, flying) and behavioural strategies (perching or patrolling) were associated with differences in thoracic temperature: patrolling males which mainly engaged in long flights and periods of basking afterwards, had lower thoracic temperatures than perching males which engaged in very short flights, fights and basking. When resting for a while thoracic temperatures did not differ between males practising different strategies. Under laboratory conditions, darker males heated up faster than pale males but there was no difference in the thoracic temperature at which they started to fly. These results indicate that thermal requirements (or general conditions) differ between the behavioural strategies, and that behavioural differences between phenotypes (colour types) relate to differences in thermal ecology. This supports the idea that darker males are better adapted to patrolling. There is no evidence that one mate-locating strategy is always superior to the other, which coincides with the observation that both strategies co-exist. More generally, this study shows that relatively small differences in colour can have a considerable effect on thermoregulation and hence on the behavioural strategies a heliothermic insect will adopt. Received: 15 August 1997 / Accepted: 15 December 1997