We thank Bryton Hixson for setup assistance. This program was supported by the Basic Research Program, Environmental Quality and Installations (EQI; Dr. Elizabeth Ferguson, Technical Director), US Army Engineer Research and Development Center.

<ol>
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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+contagion+in+mobile+animal+groups.">Behavioral contagion in mobile animal groups.</a> Proceedings of the National Academy of Sciences (U.S.A.). 112 (15), 4690-4695 (2015).</li><li>Lemasson, B. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motion+cues+tune+social+influence+in+shoaling+fish.">Motion cues tune social influence in shoaling fish.</a> Scientific Reports. 8 (1), e9785 (2018).</li><li>Kaidanovich-Beilin, O., Lipina, T., Vukobradovic, I., Roder, J., Woodgett, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+social+interaction+behaviors.">Assessment of social interaction behaviors.</a> Journal of Visualized. Experiments. (48), e2473 (2011).</li><li>Holcombe, A., Schalomon, M., Hamilton, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+method+of+drug+administration+to+multiple+zebrafish+(Danio+rerio)+and+the+quantification+of+withdrawal.">A novel method of drug administration to multiple zebrafish (Danio rerio) and the quantification of withdrawal.</a> Journal of Visualized. Experiments. (93), e51851 (2014).</li><li>Way, G. P., Southwell, M., McRobert, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Boldness,+aggression,+and+shoaling+assays+for+zebrafish+behavioral+syndromes.">Boldness, aggression, and shoaling assays for zebrafish behavioral syndromes.</a> Journal of Visualized. Experiments. (114), e54049 (2016).</li><li>Zhang, Q., Kobayashi, Y., Goto, H., Itohara, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+automated+T-maze+based+apparatus+and+protocol+for+analyzing+delay-+and+effort-based+decision+making+in+free+moving+rodents.">An automated T-maze based apparatus and protocol for analyzing delay- and effort-based decision making in free moving rodents.</a> Journal of Visualized. Experiments. (138), e57895 (2018).</li><li>Videler, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Fish Swimming. , (1993).</li><li>Orger, M. B., Smear, M. C., Anstis, S. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motion-guided+attention+promotes+adaptive+communications+during+social+navigation.">Motion-guided attention promotes adaptive communications during social navigation.</a> Proceedings of the Royal Society. 280 (1754), e20122003 (2013).</li><li>Moussa&#239;d, M., Helbing, D., Theraulaz, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=How+simple+rules+determine+pedestrian+behavior+and+crowd+disasters.">How simple rules determine pedestrian behavior and crowd disasters.</a> Proceedings of the National Academy of Sciences (U.S.A.). 108 (17), 6884-6888 (2011).</li><li>Bianco, I. H., Engert, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visuomotor+transformations+underlying+hunting+behavior+in+zebrafish).">Visuomotor transformations underlying hunting behavior in zebrafish).</a> Current Biology. 25 (7), 831-846 (2015).</li><li>Chouinard-Thuly, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technical+and+conceptual+considerations+for+using+animated+stimuli+in+studies+of+animal+behavior.">Technical and conceptual considerations for using animated stimuli in studies of animal behavior.</a> Current Zoology. 63 (1), 5-19 (2017).</li><li>Nakayasu, T., Yasugi, M., Shiraishi, S., Uchida, S., Watanabe, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+computer+graphic+animations+for+studying+social+approach+behaviour+in+medaka+fish:+Effects+of+systematic+manipulation+of+morphological+and+motion+cues.">Three-dimensional computer graphic animations for studying social approach behaviour in medaka fish: Effects of systematic manipulation of morphological and motion cues.</a> PLoS One. 12 (4), e0175059 (2017).</li><li>Stowers, J. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virtual+reality+for+freely+moving+animals.">Virtual reality for freely moving animals.</a> Nature Methods. 14 (10), 995-1002 (2017).</li><li>Warren, W. H., Kay, B., Zosh, W. D., Duchon, A. P., Sahuc, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optic+flow+is+used+to+control+human+walking.">Optic flow is used to control human walking.</a> Nature Neuroscience. 4 (2), 213-216 (2001).</li><li>Silverman, J., Suckow, M. A., Murthy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The IACUC Handbook. , (2014).</li></ol>

All authors contributed to the experimental design, analyses and writing the paper. A.C.U. and C.M.W. setup and collected the data. The authors have nothing to disclose.

Visual cues are known to trigger an optomotor response in fish exposed to black and white gratings13 and there is increasing theoretical and empirical evidence that neighbor speed plays an influential role in governing the dynamical interactions observed in fish schools7,14,15,16,17. Contrasting hypotheses exist to explain how individua...

Animals sense and interpret their habitat continuously to make informed decisions when interacting with others and navigating noisy surroundings. Individuals can enhance their situational awareness and decision making by integrating social information into their actions. Social information, however, largely stems from inference through unintended cues (i.e., sudden maneuvers to avoid a predator), which can be unreliable, rather than through direct signals that have evolved to communicate specific messages (e.g., the waggle dance in honey bees)1. Identifying how individuals rapidly assess the value of social cues, or any sensory information, can be a challenging task for investigators, particularly when individuals are traveling in groups. Vision plays an important role in governing social interactions2,3,4 and studies have inferred the interaction networks that may arise in fish schools based on each individual&#8217;s field of view5,6. Fish schools are dynamic systems, however, making it difficult to isolate individual responses to particular features, or neighbor behaviors, due to the inherent collinearities and confounding factors that arise from the interactions among group members. The purpose of this protocol is to complement current work by isolating how alternative visual features can influence the directional decisions of individuals traveling alone or within groups.
The benefit of the current protocol is to combine a manipulative experiment with computer visualization techniques to isolate the elementary visual features an individual may experience in nature. Specifically, the Y-maze (Figure 1) is used to collapse directional choice to a binary response and introduce computer animated images designed to mimic the swimming behaviors of virtual neighbors. These images are projected up from below the maze to mimic the silhouettes of conspecifics swimming beneath one or more subjects. The visual characteristics of these silhouettes, such as their morphology, speed, coherency, and swimming behavior are easily tailored to test alternative hypotheses7.
This paper demonstrates the utility of this approach by isolating how individuals of a model social fish species, the Golden Shiner (Notemigonus crysoleucas), respond to the relative speed of virtual neighbors. The protocol focus, here, is on whether the directional influence of virtual neighbors change with their speed and, if so, quantifying the form of the observed relationship. In particular, the directional cue is generated by having a fixed proportion of the silhouettes act as leaders and move ballistically towards one arm or another. The remaining silhouettes act as distractors by moving about at random to provide background noise that can be tuned by adjusting the leader/distractor ratio. The ratio of leaders to distractors captures the coherency of the directional cues and can be adjusted accordingly. Distractor silhouettes remain confined to the decision area (&#8220;DA&#8221;, Figure 1A) by having the silhouettes reflect off of the boundary. Leader silhouettes, however, are allowed to leave the DA region and enter their designated arm before slowly fading away once the silhouettes traversed 1/3 the length of the arm. As leaders leave the DA, new leader silhouettes take their place and retrace their exact path to ensure that the leader/distractor ratio remains constant in the DA throughout the experiment.
The use of virtual fish allows for the control of the visual sensory information, while monitoring the directional response of the subject, which may reveal novel features of social navigation, movement, or decision making in groups. The approach used here can be applied to a broad range of questions, such as effects of sublethal stress or predation on social interactions, by manipulating the computer animation to produce behavioral patterns of varying complexity.

<table>
	<tbody>
		<tr>
			<td>Black and white IP camera</td>
			<td>Noldus, Leesburg, VA, USA</td>
			<td></td>
			<td>https://www.noldus.com/</td>
		</tr>
		<tr>
			<td>Extruded aluminum</td>
			<td>80/20 Inc., Columbia City, IN, USA</td>
			<td>3030-S</td>
			<td>https://www.8020.net 3.00&#34; X 3.00&#34; Smooth T-Slotted Profile, Eight Open T-Slots</td>
		</tr>
		<tr>
			<td>Finfish Starter with Vpak, 1.5 mm extruded pellets</td>
			<td>Zeigler Bros. Inc., Gardners, PA, USA</td>
			<td></td>
			<td>http://www.zeiglerfeed.com/</td>
		</tr>
		<tr>
			<td>Golden shiners</td>
			<td>Saul Minnow Farm, AR, USA</td>
			<td></td>
			<td>http://saulminnow.com/</td>
		</tr>
		<tr>
			<td>ImageJ (v 1.52h) freeware</td>
			<td>National Institute for Health (NIH), USA</td>
			<td></td>
			<td>https://imagej.nih.gov/ij/</td>
		</tr>
		<tr>
			<td>LED track lighting</td>
			<td>Lithonia Lightening, Conyers, GA, USA</td>
			<td>BR20MW-M4</td>
			<td>https://lithonia.acuitybrands.com/residential-track</td>
		</tr>
		<tr>
			<td>Oracle 651 white cut vinyl</td>
			<td>651Vinyl, Louisville, KY, USA</td>
			<td>651-010M-12:5ft</td>
			<td>http://www.651vinyl.com. Can order various sizes.</td>
		</tr>
		<tr>
			<td>PowerLite 570 overhead projector</td>
			<td>Epson, Long Beach CA, USA</td>
			<td>V11H605020</td>
			<td>https://epson.com/For-Work/Projectors/Classroom/PowerLite-570-XGA-3LCD-Projector/p/V11H605020</td>
		</tr>
		<tr>
			<td>Processing (v 3) freeware</td>
			<td>Processing Foundation</td>
			<td></td>
			<td>https://processing.org/</td>
		</tr>
		<tr>
			<td>R (3.5.1) freeware</td>
			<td>The R Project for Statistical Computing</td>
			<td></td>
			<td>https://www.r-project.org/</td>
		</tr>
		<tr>
			<td>Ultra-white 360 theater screen</td>
			<td>Alternative Screen Solutions, Clinton, MI, USA</td>
			<td>1950</td>
			<td>https://www.gooscreen.com. Must call for special cut size</td>
		</tr>
		<tr>
			<td>Z-Hab system</td>
			<td>Pentair Aquatic Ecosystems, Apopka, FL, USA</td>
			<td></td>
			<td>https://pentairaes.com/. Call for details and sizing.</td>
		</tr>
	</tbody>
</table>

integrating visual psychophysical assays within a y-maze to isolate the role that visual features play in navigational decisions

Collective animal behavior arises from individual motivations and social interactions that are critical for individual fitness. Fish have long inspired investigations into collective motion, specifically, their ability to integrate environmental and social information across ecological contexts. This demonstration illustrates techniques used for quantifying behavioral responses of fish, in this case, Golden Shiner (Notemigonus crysoleucas), to visual stimuli using computer visualization and digital image analysis. Recent advancements in computer visualization allow for empirical testing in the lab where visual features can be controlled and finely manipulated to isolate the mechanisms of social interactions. The purpose of this method is to isolate visual features that can influence the directional decisions of the individual, whether solitary or with groups. This protocol provides specifics on the physical Y-maze domain, recording equipment, settings and calibrations of the projector and animation, experimental steps and data analyses. These techniques demonstrate that computer animation can elicit biologically-meaningful responses. Moreover, the techniques are easily adaptable to test alternative hypotheses, domains, and species for a broad range of experimental applications. The use of virtual stimuli allows for the reduction and replacement of the number of live animals required, and consequently reduces laboratory overhead.
This demonstration tests the hypothesis that small relative differences in the movement speeds (2 body lengths per second) of virtual conspecifics will improve the speed and accuracy with which shiners follow the directional cues provided by the virtual silhouettes. Results show that shiners directional decisions are significantly affected by increases in the speed of the visual cues, even in the presence of background noise (67% image coherency). In the absence of any motion cues, subjects chose their directions at random. The relationship between decision speed and cue speed was variable and increases in cue speed had a modestly disproportionate influence on directional accuracy.

Hypothesis and design 
To demonstrate the utility of this experimental system we tested the hypothesis that the accuracy with which Golden Shiner follow a visual cue will improve with the speed of that cue. Wild type Golden Shiner were used (N = 16, body lengths, BL, and wet weights, WW, were 63.4 &#177; 3.5 mm and 1.8 &#177; 0.3 g, respectfully). The coherency of the visual stimuli (leader/distractor r...

Watch this Scientific Journal Video about Integrating Visual Psychophysical Assays within a Y-Maze to Isolate the Role that Visual Features Play in Navigational Decisions at JoVE.com

Integrating Visual Psychophysical Assays within a Y-Maze to Isolate the Role that Visual Features Play in Navigational Decisions

Here, we present a protocol to demonstrate a behavioral assay that quantifies how alternative visual features, such as motion cues, influence directional decisions in fish. Representative data are presented on the speed and accuracy where Golden Shiner (Notemigonus crysoleucas) follow virtual fish movements.

Collective animal behavior arises from individual motivations and social interactions that are critical for individual fitness. Fish have long inspired ...

integrating-visual-psychophysical-assays-within-y-maze-to-isolate

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SCIENTISTS IN ACTION

6.0K vistas.  US Army Engineer Research and Development Center. Nuestro protocolo nos permite imitar las características visuales que un pez que se mueve libremente puede experimentar cuando viaja en grupo y cuantificar cómo esas características influyen en sus decisiones de navegación. Esta técnica se puede utilizar para vincular las decisiones direccionales a nivel individual en respuesta a una señal sensorial para ver cómo cambian esas decisiones en función de las acciones de otros. Nuestros métodos proporcionan información sobre cómo los an...

Video: Integrating Visual Psychophysical Assays within a Y-Maze to Isolate the Role that Visual Features Play in Navigational Decisions

Methods to Explore the Influence of Top-down Visual Processes on Motor Behavior

Kinesthetic awareness is important to successfully navigate the environment. When we interact with our daily surroundings, some aspects of movement are deliberately planned, while others spontaneously occur below conscious awareness. The deliberate component of this dichotomy has been studied extensively in several contexts, while the spontaneous component remains largely under-explored. Moreover, how perceptual processes modulate these movement classes is still unclear. In particular, a currently debated issue is whether the visuomotor system is governed by the spatial percept produced by a visual illusion or whether it is not affected by the illusion and is governed instead by the veridical percept. Bistable percepts such as 3D depth inversion illusions (DIIs) provide an excellent context to study such interactions and balance, particularly when used in combination with reach-to-grasp movements. In this study, a methodology is developed that uses a DII to clarify the role of top-down processes on motor action, particularly exploring how reaches toward a target on a DII are affected in both deliberate and spontaneous movement domains.

It is unclear how top-down signals from the ventral visual stream affect movement. We developed a paradigm to test motor behavior towards a target on a 3D depth inversion illusion. Significant differences are reported in both deliberate, goal-directed movements and automatic actions under illusory and veridical viewing conditions.

Kinesthetic awareness is important to successfully navigate the environment. When we interact with our daily surroundings, some aspects of movement are ...

Practical Methodology of Cognitive Tasks Within a Navigational Assessment

This paper describes an approach for measuring navigation accuracy relative to cognitive skills. The methodology behind the assessment will thus be clearly outlined in a step-by-step manner. Navigational skills are important when trying to find symbols within a speech-generating device (SGD) that has a dynamic screen and taxonomical organization. The following skills have been found to impact children&#8217;s ability to find symbols when navigating within the levels of an SGD: sustained attention, categorization, cognitive flexibility, and fluid reasoning1,2. According to past studies, working memory was not correlated with navigation1,2.
The materials needed for this method include a computerized tablet, an augmentative and alternative communication application, a booklet of symbols, and the Leiter International Performance Scale-Revised (Leiter-R)3. This method has been used in two previous studies. Robillard, Mayer-Crittenden, Roy-Charland, Minor-Corriveau and B&#233;langer1 assessed typically developing children, while Rondeau, Robillard and Roy-Charland2 assessed children and adolescents with a diagnosis of Autism Spectrum Disorder. The direct observation of this method will facilitate the replication of this study for researchers. It will also help clinicians that work with children who have complex communication needs to determine the children&#8217;s ability to navigate an SGD with taxonomical categorization.

Children who have complex communication needs can benefit from the use of a speech-generating device (SGD). Cognitive skills were identified as having an impact on the ability to navigate between levels of SGDs. This protocol describes the steps needed for Speech-language Pathologists to assess cognitive skills and navigational abilities.

This paper describes an approach for measuring navigation accuracy relative to cognitive skills. The methodology behind the assessment will thus be ...

Methods to Test Visual Attention Online

Online data collection methods have particular appeal to behavioral scientists because they offer the promise of much larger and much more representative data samples than can typically be collected on college campuses. However, before such methods can be widely adopted, a number of technological challenges must be overcome &#8211; in particular in experiments where tight control over stimulus properties is necessary. Here we present methods for collecting performance data on two tests of visual attention. Both tests require control over the visual angle of the stimuli (which in turn requires knowledge of the viewing distance, monitor size, screen resolution, etc.) and the timing of the stimuli (as the tests involve either briefly flashed stimuli or stimuli that move at specific rates). Data collected on these tests from over 1,700 online participants were consistent with data collected in laboratory-based versions of the exact same tests. These results suggest that with proper care, timing/stimulus size dependent tasks can be deployed in web-based settings.

To replicate laboratory settings, online data collection methods for visual tasks require tight control over stimulus presentation. We outline methods for the use of a web application to collect performance data on two tests of visual attention.

Online data collection methods have particular appeal to behavioral scientists because they offer the promise of much larger and much more representative ...

A Method to Quantify Visual Information Processing in Children Using Eye Tracking

Visual problems that occur early in life can have major impact on a child's development. Without verbal communication and only based on observational methods, it is difficult to make a quantitative assessment of a child's visual problems. This limits accurate diagnostics in children under the age of 4 years and in children with intellectual disabilities. Here we describe a quantitative method that overcomes these problems. The method uses a remote eye tracker and a four choice preferential looking paradigm to measure eye movement responses to different visual stimuli. The child sits without head support in front of a monitor with integrated infrared cameras. In one of four monitor quadrants a visual stimulus is presented. Each stimulus has a specific visual modality with respect to the background, e.g., form, motion, contrast or color. From the reflexive eye movement responses to these specific visual modalities, output parameters such as reaction times, fixation accuracy and fixation duration are calculated to quantify a child's viewing behavior. With this approach, the quality of visual information processing can be assessed without the use of communication. By comparing results with reference values obtained in typically developing children from 0-12 years, the method provides a characterization of visual information processing in visually impaired children. The quantitative information provided by this method can be advantageous for the field of clinical visual assessment and rehabilitation in multiple ways. The parameter values provide a good basis to: (i) characterize early visual capacities and consequently to enable early interventions; (ii) compare risk groups and follow visual development over time; and (iii), construct an individual visual profile for each child.

A method is described to quantify the quality of visual information processing based on reflexive eye movements in response to specific visual modalities. Reaction times and fixation output parameters are used to characterize visual performance in children with and without visual impairments from 6 months of age.

Visual problems that occur early in life can have major impact on a child's development. Without verbal communication and only based on observational ...

Visualizing Visual Adaptation

Many techniques have been developed to visualize how an image would appear to an individual with a different visual sensitivity: e.g., because of optical or age differences, or a color deficiency or disease. This protocol describes a technique for incorporating sensory adaptation into the simulations. The protocol is illustrated with the example of color vision, but is generally applicable to any form of visual adaptation. The protocol uses a simple model of human color vision based on standard and plausible assumptions about the retinal and cortical mechanisms encoding color and how these adjust their sensitivity to both the average color and range of color in the prevailing stimulus. The gains of the mechanisms are adapted so that their mean response under one context is equated for a different context. The simulations help reveal the theoretical limits of adaptation and generate &#34;adapted images&#34; that are optimally matched to a specific environment or observer. They also provide a common metric for exploring the effects of adaptation within different observers or different environments. Characterizing visual perception and performance with these images provides a novel tool for studying the functions and consequences of long-term adaptation in vision or other sensory systems.

This article describes a novel method for simulating and studying adaptation in the visual system.

Many techniques have been developed to visualize how an image would appear to an individual with a different visual sensitivity: e.g., because of optical ...

Using Eye Movements Recorded in the Visual World Paradigm to Explore the Online Processing of Spoken Language

In a typical eye tracking study using the visual world paradigm, participants' eye movements to objects or pictures in the visual workspace are recorded via an eye tracker as the participant produces or comprehends a spoken language describing the concurrent visual world. This paradigm has high versatility, as it can be used in a wide range of populations, including those who cannot read and/or who cannot overtly give their behavioral responses, such as preliterate children, elderly adults, and patients. More importantly, the paradigm is extremely sensitive to fine grained manipulations of the speech signal, and it can be used to study the online processing of most topics in language comprehension at multiple levels, such as the fine grained acoustic phonetic features, the properties of words, and the linguistic structures. The protocol described in this article illustrates how a typical visual world eye tracking study is conducted, with an example showing how the online processing of some semantically complex statements can be explored with the visual world paradigm.

The visual world paradigm monitors participants' eye movements in the visual workspace as they are listening to or speaking a spoken language. This paradigm can be used to investigate the online processing of a wide range of psycholinguistic questions, including semantically complex statements, such as disjunctive statements.

In a typical eye tracking study using the visual world paradigm, participants' eye movements to objects or pictures in the visual workspace are recorded ...

Visual Classical Conditioning in Wood Ants

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, studies implementing classical conditioning paradigms with harnessed insects have been important for investigating the exact cues that individuals learn and the neural mechanisms underlying learning and memory formation. Here we present a protocol for evoking visual associative learning in wood ants through classical conditioning. In this paradigm, ants are harnessed and presented with a visual cue (a blue cardboard), the conditional stimulus (CS), paired with an appetitive sugar reward, the unconditional stimulus (US). Ants perform a Maxilla-Labium Extension Reflex (MaLER), the unconditional response (UR), which can be used as a readout for learning. Training consists of 10 trials, separated by a 5-minute intertrial interval (ITI). Ants are also tested for memory retention 10 minutes or 1 hour after training. This protocol has the potential to allow researchers to analyze, in a precise and controlled manner, the details of visual memory formation and the neural basis of learning and memory formation in wood ants.

We present a protocol for the classical conditioning of harnessed ants that permits researchers to study visual learning and memory formation at a level of analysis not possible with freely moving individuals.

Several species of insects have become model systems for studying learning and memory formation. Although many studies focus on freely moving animals, ...

A Visual Guide for Studying Behavioral Defenses to Pathogen Attacks in Leaf-Cutting Ants

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies have described the behaviors connected with disease defense, and the associated use of chemicals and antimicrobials, no common visual reference has been made. The main aim of this study was to record short clips of behaviors involved in disease defense, both prophylactically and directly targeted towards an antagonist of the colony following infection. To do so we used an infection experiment, with sub-colonies of the leaf-cutting ant species Acromyrmex echinatior, and the most significant known pathogenic threat to the ants&#39; fungal crop (Leucoagaricus gongylophorus), a specialized pathogenic fungus in the genus Escovopsis. We filmed and compared infected and uninfected colonies, at both early and more advanced stages of infection. We quantified key defensive behaviors across treatments and show that the behavioral response to pathogen attack likely varies between different castes of worker ants, and between early and late detection of a threat. Based on these recordings we have made a library of behavioral clips, accompanied by definitions of the main individual defensive behaviors. We anticipate that such a guide can provide a common frame of reference for other researchers working in this field, to recognize and study these behaviors, and also provide greater scope for comparing different studies to ultimately help better understand the role these behaviors play in disease defense.

We present a visual guide to disease defense behaviors in leaf-cutting ants, with individual clips and accompanying definitions, illustrated in an experimental infection scenario. Our main aim is to help other researchers recognize key defensive behaviors and to provide a common understanding for future research in this field.

The complex lifestyle, evolutionary history of advanced cooperation, and disease defenses of leaf-cutting ants are well studied. Although numerous studies ...

Using the Visual World Paradigm to Study Sentence Comprehension in Mandarin-Speaking Children with Autism

Sentence comprehension relies on the ability to rapidly integrate different types of linguistic and non-linguistic information. However, there is currently a paucity of research exploring how preschool children with autism understand sentences using different types of cues. The mechanisms underlying sentence comprehension remains largely unclear. The present study presents a protocol to examine the sentence comprehension abilities of preschool children with autism. More specifically, a visual world paradigm of eye-tracking is used to explore the moment-to-moment sentence comprehension in the children. The paradigm has multiple advantages. First, it is sensitive to the time course of sentence comprehension and thus can provide rich information about how sentence comprehension unfolds over time. Second, it requires minimal task and communication demands, so it is ideal for testing children with autism. To further minimize the computational burden of children, the present study measures eye movements that arise as automatic responses to linguistic input rather than measuring eye movements that accompany conscious responses to spoken instructions.

We present a protocol to examine the use of morphological cues during real-time sentence comprehension by children with autism.

Sentence comprehension relies on the ability to rapidly integrate different types of linguistic and non-linguistic information. However, there is ...

Exploring Infant Sensitivity to Visual Language using Eye Tracking and the Preferential Looking Paradigm

We discuss the use of the preferential looking paradigm in eye tracking studies in order to study how infants develop, understand, and attend to the world around them. Eye tracking is a safe and non-invasive way to collect gaze data from infants, and the preferential looking paradigm is simple to design and only requires the infant to be attending to the screen. By simultaneously showing two visual stimuli that differ in one dimension, we can assess whether infants show different looking behavior for either stimulus, thus demonstrating sensitivity to that difference. The challenges in such experimental approaches are that experiments must be kept brief (no more than 10 min) and be carefully controlled such that the two stimuli differ in only one way. The interpretation of null results must also be carefully considered. In this paper, we illustrate a successful example of an infant eye tracking study with a preferential looking paradigm to discover that 6-month-olds are sensitive to linguistic cues in a signed language despite having no prior exposure to signed language, suggesting that infants possess intrinsic or innate sensitivities to these cues.

Eye tracking studies using a preferential looking paradigm can be used to study infants&#39;&#160;emerging understanding of, and attention to, their external visual world.

We discuss the use of the preferential looking paradigm in eye tracking studies in order to study how infants develop, understand, and attend to the world ...