Runway extinction in terrestrial toads (Rhinella arenarum): Instrumental or Pavlovian?

Instrumental appetitive extinction involves the reduction of a previously reinforced response when its occurrence is no longer rewarded. Two experiments with terrestrial toads (Rhinella arenarum) tested whether the occurrence of a nonreinforced response is necessary for response extinction by varying the time of exposure to nonrewarded goal-box stimuli across groups. In Experiment 1, toads that received the same acquisition training (15 sessions, 1 session/day, 300 s of access to water in the goal box) were randomly assigned to two groups. In Group 600 (n=12), animals spent 600 s in the goal box in 8 daily extinction sessions (water present but inaccessible). In Group 0 (n=11), toads performed the runway response (i.e., walking from the start to the goal box) but were removed as soon as they entered the goal box, thus having minimal exposure to nonrewarded goal-box stimuli. The runway response was weakened in Group 600 across extinction trials, but exhibited little change in Group 0. In Experiment 2, toads were randomly assigned to two groups after the same acquisition training. Group 0 (n=7) was treated the same as Group 0 in the previous experiment. In Group RI (retention interval, n=7), toads remained in their home cage for 13 days. Finally, all animals received 4 extinction sessions with 300 s in the empty goal box. There was little behavioral change in Group 0 during the 13 sessions with minimal exposure to the goal box. In extinction, both groups reduced their runway response at similar rates. Although the procedures were instrumental, extinction of the runway response in toads can be accounted for in terms of a Pavlovian approach response to stimuli paired with reward and nonreward in the goal box.

Amphibian spatial cognition, medial pallium and other supporting telencephalic structures.

Vertebrate hippocampal formation is central to conversations on the comparative analysis of spatial cognition, especially in light of variation found in different vertebrate classes. Assuming the medial pallium (MP) of extant amphibians resembles the hippocampal formation (HF) of ancestral stem tetrapods, we propose that the HF of modern amniotes began with a MP characterized by a relatively undifferentiated cytoarchitecture, more direct thalamic/olfactory sensory inputs, and a more generalized role in associative learning-memory processes. As such, hippocampal evolution in amniotes, especially mammals, can be seen as progressing toward a cytoarchitecture with well-defined subdivisions, regional connectivity, and a functional specialization supporting map-like representations of space. We then summarize a growing literature on amphibian spatial cognition and its underlying brain organization. Emphasizing the MP/HF, we highlight that further research into amphibian spatial cognition would provide novel insight into the role of the HF in spatial memory processes, and their supporting neural mechanisms. A more complete reconstruction of hippocampal evolution would benefit from additional research on non-mammalian vertebrates, with amphibians being of particular interest.

Instrumental successive negative contrast in rats: Trial distribution, reward magnitude, and prefrontal cortex activation.

Successive negative contrast (SNC) has been used to study reward relativity, reward loss, and frustration for decades. In instrumental SNC (iSNC), the anticipatory performance of animals downshifted from a large reward to a small reward is compared to that of animals always reinforced with the small reward. iSNC involves a transient deterioration of anticipatory behavior in downshifted animals compared to unshifted controls. There is scattered information on the optimal parameters to produce this effect and even less information about its neural basis. Five experiments with rats trained in a runway to collect food pellets explored the effects of trial distribution (massed or spaced), amount of preshift training, reward disparity, and reward magnitude on the development of an iSNC effect. Start, run, and goal latencies were measured. Using spaced trials (one trial per day), evidence of the iSNC effect was observed with 24 preshift trials and a 32-to-4 pellet disparity. With massed trials (4 trials per session separated by 30-s intertrial intervals), evidence of iSNC was found with 12 preshift sessions (a total of 48 trials) and a 16-to-2 pellet disparity. The massed-training procedure was then used to assess neural activity in three prefrontal cortex areas using c-Fos expression in animals perfused after the first downshift session. There was evidence of increased activation in the anterior cingulate cortex and a trend toward increased activation in the infralimbic and prelimbic cortices. These procedures open a venue for studying the neural basis of the instrumental behavior of animals that experience reward loss.

The medial pallium and the spatial encoding of geometric and visual cues in the terrestrial toad, Rhinella arenarum.

The medial pallium (MP) of amphibians is the homologue of the mammalian hippocampus, and previous research has implicated MP for locating a using the boundary geometry of an environment. MP-lesioned, sham-operated and intact control terrestrial toads, Rhinella arenarum, were trained to locate a goal in a rectangular arena with a visual feature cue placed on one of the short walls. Whereas the sham-operated and intact subjects successfully learned to locate the goal, the MP-lesioned toads showed no evidence of learning. The data support the hypothesis that the amphibian MP is involved when the boundary geometry of an environment is used to locate a goal, which is consistent with evidence from other vertebrate groups. Curious, however, is that the MP lesions also resulted in the toads' inability to locate the goal based on the visual feature cue. This result supports previous research and suggests that, in contrast to the hippocampal homologue of amniotes, the amphibian medial pallium plays a broader role in spatial learning processes.

Evidence of successive negative contrast in terrestrial toads (Rhinella arenarum): central or peripheral effect?

Prior research with terrestrial toads (Rhinella arenarum) in a water-reinforced instrumental situation indicated a direct relationship between acquisition rate and reward magnitude. However, a reward downshift produced a gradual adjustment of instrumental performance and a rapid adjustment of consummatory performance, rather than the abrupt and transient deterioration of behavior typical of a successive negative contrast effect. In Experiment 1, using a two-chamber box, a downshift from deionized water (which supports maximal rehydration) to 250-mM sodium chloride solution (which supports a lower rehydration), also yielded a gradual adjustment of instrumental behavior. In this experiment, animals received one trial per day and were allowed 300 s of access to the reward in the goal box. Experiment 2 used the same procedure, except that animals were allowed access to the solution in the goal box for 600 s. Under these conditions, reward downshift led to longer latencies (instrumental) and lower rehydration levels (consummatory) than those of unshifted controls, providing evidence for successive negative contrast. Unlike in similar experiments with mammals, the effect was not transient, but persisted relatively unmodified over twelve daily postshift trials. In this case, the possibility of adaptation of the peripheral mechanisms for water uptake is considered. The comparative relevance of these results is discussed in terms of habit formation versus expectancy-guided behavior in vertebrate learning.

Perceived Concerns and Psychological Distress of Healthcare Workers Facing Three Early Stages of COVID-19 Pandemic.

Background: This study analyzed the difference in psychological distress of the healthcare workers in three different periods of the coronavirus disease 2019 (COVID-19) pandemic in Argentina. Specifically, from the third week of the mandatory quarantine through the two following weeks. Methods: Analysis of the responses of 1,458 members of the health personnel was done on a questionnaire on healthcare workers concerns regarding the care of patients with coronavirus, indicators of depression, anxiety, intolerance of uncertainty, and coping. Results: The psychological indicators that were considered presented differences between the evaluated periods. Perceived concerns about the possibility of infecting loved ones and infecting themselves were greatest in the periods after the onset of the pandemic. In addition, the perception of how the work environment worsened and how lack of sleep interfered with their work was also higher in periods 2 and 3. The same results were found in the indicators of depression, anxiety, and intolerance of uncertainty. Finally, the indicators of high tension and concurrent lack of emotional control, which was greater in the last periods evaluated, were also expressed in the coping strategies (showing emotional lability, only contained by hypercontrol). Conclusion: The differences found in the psychological indicators between the evaluated periods support the need for early psychological care of health personnel which should be a priority of public health and a fundamental fact to increase its immediate effectiveness in the care of infected patients.

On the transfer of spatial learning between geometrically different shaped environments in the terrestrial toad, Rhinella arenarum.

When trained in a rectangular arena, some research has suggested that rats are guided by local features rather than overall boundary geometry. We explored this hypothesis using the terrestrial toad, Rhinella arenarum, as a comparative contrast. In two experiments, toads were trained to find a water-reward goal location in either a featureless rectangular arena (Experiment 1) or in a rectangular arena with a removable colored feature panel covering one short wall (Experiment 2). After learning to successfully locate the water reward, probe trials were carried out by changing the shape of the arena into a kite form with two 90°-angled corners, and in the case of Experiment 2, also shifting the location of the color panel. The results of Experiment 1 indicated that the toads, in contrast to rats, relied primarily on overall shape or boundary geometry to encode the location of a goal. Under the probe conditions of the altered environmental geometry in Experiment 2, the toads seemed to preferentially choose a corner that was generally correct relative to the feature panel experienced during training. Together, the data of the current study suggest that toads and rats differ in the strategies they employ to represent spatial information available in a rectangular arena. Further, the results support the hypothesis that amphibians and mammals engage different neural mechanisms, perhaps related to different evolutionary selective pressures, for the representation of environmental geometry used for navigation.

The Mating Call of the Terrestrial Toad, Rhinella arenarum, as a Cue for Spatial Orientation and Its Associated Brain Activity.

Acoustic communication is essential for reproduction and predator avoidance in many anuran species. For example, mating calls are generally produced by males and represent a conspicuous communication signal employed during the breeding season. Although anuran mating calls have been largely studied to analyze content and phonotaxis toward choruses, they are rarely discussed as sources of information guiding spatial behavior in broader contexts. This is striking if we consider that previous studies have shown anurans to be impressive navigators. In the current study, we investigated whether terrestrial toad (Rhinella arenarum) males can use a mating call as a spatial cue to locate a water reward in a laboratory maze. Male toads could indeed learn the location of a reward guided by a mating call. This navigational ability, as indicated by c-Fos, was associated with greater neuronal activity in the telencephalic hippocampal formation (HF; also referred to in amphibians as medial pallium), the medial septum (MS), and the central amygdala (CeA). HF and MS are telencephalic structures associated with spatial navigation in mammals and other vertebrates. The CeA, by contrast, has been studied in the context of acoustic processing and communication in other amphibian species. The results are discussed in the framework of an evolutionary conserved, HF-septal spatial-cognitive network shared by amphibians and mammals.

Local geometric properties do not support reorientation in hippocampus-engaged homing pigeons.

It is generally accepted that the geometry of an environment is a reliable source of information for spatial navigation used by most vertebrate species. However, there is a continuing debate on which geometrical properties of space are the ones that matter for reorientation. In this study, pigeons were trained to find a food reward hidden in 2 opposite corners in a rectangular arena. The animals were then tested in a kite-shaped environment similar to Pearce, Good, Jones, and McGregor (2004). We found that pigeons, unlike rats, were not able to identify the correct corner in the kite arena even though elements clearly preserved the correct long wall-short wall geometric configuration and the local aspect of the trained goal. This behavioral study was followed by a c-Fos, IEG analysis of brain activation that contrasted pigeons exposed to the trained, familiar rectangular environment with pigeons that were exposed to an unfamiliar, trapezoid arena. The hippocampal formation (HF) displayed greater c-Fos expression in the animals exposed to the familiar, training arena, which further supports the conclusion that pigeons do not substantially rely on local geometric features for reorientation. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Duration of extinction trials as a determinant of instrumental extinction in terrestrial toads (Rhinella arenarum).

Instrumental learning guides behavior toward resources. When such resources are no longer available, approach to previously reinforced locations is reduced, a process called extinction. The present experiments are concerned with factors affecting the extinction of acquired behaviors in toads. In previous experiments, total reward magnitude in acquisition and duration of extinction trials were confounded. The present experiments were designed to test the effects of these factors in factorial designs. Experiment 1 varied reward magnitude (900, 300, or 100 s of water access per trial) and amount of acquisition training (5 or 15 daily trials). With total amount of water access equated in acquisition, extinction with large rewards was faster (longer latencies in 900/5 than 300/15), but with total amount of training equated, extinction with small rewards was faster (longer latencies in 100/15 than 300/15). Experiment 2 varied reward magnitude (1200 or 120 s of water access per trial) while holding constant the number of acquisition trials (5 daily trials) and the duration of extinction trials (300 s). Extinction performance was lower with small, rather than large reward magnitude (longer latencies in 120/300 than in 1200/300). Thus, instrumental extinction depends upon the amount of time toads are exposed to the empty goal compartment during extinction trials.

Reflections on the Structural-Functional Evolution of the Hippocampus: What Is the Big Deal about a Dentate Gyrus?

The vertebrate hippocampal formation has been central in discussions of comparative cognition, nurturing an interest in understanding the evolution of variation in hippocampal organization among vertebrate taxa and the functional consequences of that variation. Assuming some similarity between the medial pallium of extant amphibians and the hippocampus of stem tetrapods, we propose the hypothesis that the hippocampus of modern amniotes began with a medial pallium characterized by a relatively undifferentiated cytoarchitecture, more direct thalamic and olfactory sensory inputs, and a broad role in associative learning and memory processes that nonetheless included the map-like representation of space. From this modest beginning evolved the cognitively more specialized hippocampal formation of birds and the hippocampus of mammals with its confounding dentate gyrus. Much has been made of trying to identify a dentate homologue in birds, but there are compelling reasons to believe no such structural homologue/functional equivalent exists. The uniqueness of the mammalian dentate then raises the question of what might be the functional consequences of a hippocampus with a dentate compared to one without. One might be tempted to speculate that the presence of a dentate gyrus facilitates so-called pattern separation, but birds with their suspected dentate-less hippocampus display excellent hippocampal-dependent pattern separation relying on space. Perhaps one consequence of a dentate is a hippocampus better designed to process a broader array of stimuli beyond space to more robustly support episodic memory. What is clear is that any meaningful reconstruction of hippocampal evolution and the eventual identification of any subdivisional homologies will require more data on the neurobiological and functional properties of the nonmammalian hippocampus, particularly those of amphibians and reptiles.

Slope-based and geometric encoding of a goal location by the terrestrial toad (Rhinella arenarum).

The current study was designed to test for the ability of terrestrial toads, Rhinella arenarum, to use slope as source of spatial information to locate a goal, and investigate the relative importance of slope and geometric information for goal localization. Toads were trained to locate a single, water-reward goal location in a corner of a rectangular arena placed on an incline. Once the toads learned the task, 3 types of probe trials were carried out to determine the relative use of slope and geometric information for goal localization. The probe trials revealed that the toads were able to independently use slope, and as previously reported, geometry to locate the goal. However, the boundary geometry of the experimental arena was found to be preferentially used by the toads when geometric and slope information were set in conflict. (PsycINFO Database Record

Telencephalic Neuronal Activation Associated with Spatial Memory in the Terrestrial Toad Rhinella arenarum: Participation of the Medial Pallium during Navigation by Geometry.

Amphibians are central to discussions of vertebrate evolution because they represent the transition from aquatic to terrestrial life, a transition with profound consequences for the selective pressures shaping brain evolution. Spatial navigation is one class of behavior that has attracted the interest of comparative neurobiologists because of the relevance of the medial pallium/hippocampus, yet, surprisingly, in this regard amphibians have been sparsely investigated. In the current study, we trained toads to locate a water goal relying on the boundary geometry of a test environment (Geometry-Only) or boundary geometry coupled with a prominent, visual feature cue (Geometry-Feature). Once learning had been achieved, the animals were given one last training session and their telencephali were processed for c-Fos activation. Compared to control toads exposed to the test environment for the first time, geometry-only toads were found to have increased neuronal labeling in the medial pallium, the presumptive hippocampal homologue, while geometry-feature toads were found to have increased neuronal labeling in the medial, dorsal, and lateral pallia. The data indicate medial pallial participation in guiding navigation by environmental geometry and lateral, and to a lesser extent dorsal, pallial participation in guiding navigation by a prominent visual feature. As such, participation of the medial pallium/hippocampus in spatial cognition appears to be a conserved feature of terrestrial vertebrates even if their life history is still tied to water, a brain-behavior feature seemingly at least as ancient as the evolutionary transition to life on land.

Telencephalic neural activation following passive avoidance learning in a terrestrial toad.

The present study explores passive avoidance learning and its neural basis in toads (Rhinella arenarum). In Experiment 1, two groups of toads learned to move from a lighted compartment into a dark compartment. After responding, animals in the experimental condition were exposed to an 800-mM strongly hypertonic NaCl solution that leads to weight loss. Control animals received exposure to a 300-mM slightly hypertonic NaCl solution that leads to neither weight gain nor loss. After 10 daily acquisition trials, animals in the experimental group showed significantly longer latency to enter the dark compartment. Additionally, 10 daily trials in which both groups received the 300-mM NaCl solution after responding eliminated this group effect. Thus, experimental animals showed gradual acquisition and extinction of a passive avoidance respond. Experiment 2 replicated the gradual acquisition effect, but, after the last trial, animals were sacrificed and neural activation was assessed in five brain regions using AgNOR staining for nucleoli-an index of brain activity. Higher activation in the experimental animals, relative to controls, was observed in the amygdala and striatum. Group differences in two other regions, lateral pallium and septum, were borderline, but nonsignificant, whereas group differences in the medial pallium were nonsignificant. These preliminary results suggest that a striatal-amygdala activation could be a key component of the brain circuit controlling passive avoidance learning in amphibians. The results are discussed in relation to the results of analogous experiments with other vertebrates.

Sex differences in retention after a visual or a spatial discrimination learning task in brood parasitic shiny cowbirds.

Females of avian brood parasites, like the shiny cowbird (Molothrus bonariensis), locate host nests and on subsequent days return to parasitize them. This ecological pressure for remembering the precise location of multiple host nests may have selected for superior spatial memory abilities. We tested the hypothesis that shiny cowbirds show sex differences in spatial memory abilities associated with sex differences in host nest searching behavior and relative hippocampus volume. We evaluated sex differences during acquisition, reversal and retention after extinction in a visual and a spatial discrimination learning task. Contrary to our prediction, females did not outperform males in the spatial task in either the acquisition or the reversal phases. Similarly, there were no sex differences in either phase in the visual task. During extinction, in both tasks the retention of females was significantly higher than expected by chance up to 50 days after the last rewarded session (∼85-90% of the trials with correct responses), but the performance of males at that time did not differ than that expected by chance. This last result shows a long-term memory capacity of female shiny cowbirds, which were able to remember information learned using either spatial or visual cues after a long retention interval.

Use of local visual cues for spatial orientation in terrestrial toads (Rhinella arenarum): The role of distance to a goal.

The use of environmental visual cues for navigation is an ability present in many groups of animals. The effect of spatial proximity between a visual cue and a goal on reorientation in an environment has been studied in several vertebrate groups, but never previously in amphibians. In this study, we tested the use of local visual cues (beacons) to orient in an open field in the terrestrial toad (Rhinella arenarum). Experiment 1 showed that toads could orient in space using 2 cues located near the rewarded container. Experiment 2 used only 1 cue placed at different distances to the goal and revealed that learning speed was affected by the proximity to the goal (the closer the cue was to the goal, the faster toads learned its location). Experiment 3 showed that the position of a cue results in a different predictive value. Toads preferred cues located closer to the goal more than those located farther away as a reference for orientation. Present results revealed, for the first time, that (a) toads can learn to orient in an open space using visual cues, and that (b) the effect of spatial proximity between a cue and a goal, a learning phenomenon previously observed in other groups of animals such as mammals, birds, fish, and invertebrates, also affects orientation in amphibians. Thus, our results suggest that toads are able to employ spatial strategies that closely parallel those described in other vertebrate groups, supporting an early evolutionary origin for these spatial orientation skills.

The role of stimulus complexity, age and experience in the expression of exploratory behaviour in the Chimango Caracara, Milvago chimango.

Exploration represents an important way by which organisms evaluate environment information. The decision of whether or not an animal should investigate environmental changes may influence the extent to which animals learn about their surroundings and cope with habitat modifications. We analysed exploration behaviour in a suburban population of a raptor species, the Chimango Caracara, Milvago chimango, by examining how age, previous experience and object complexity influence novel object exploration. Our findings showed that object complexity did not influence caracaras initial approach and contact with objects, but did influence the degree of engagement during exploratory activities, as measured by total exploration time and number of exploration events. These variables were higher for complex objects than for simple objects. Experience resulted in less exploration of simple objects. It is likely that, for caracaras, simple objects are easier to encode and recall than complex objects, so additional exploration of such objects would not provide further information. Results suggest that exploratory behaviour in this raptor was guided more by the benefits of a greater quantity of information obtained by exploring complex objects, than by the risks associated to this activity. We can conclude that caracaras cope with novel features in their surroundings with a novelty-seeking strategy, characteristic for generalist species in discovering early new resources opportunities, and which might be a determining factor for adaptive responses to environment modification.

Goal orientation by geometric and feature cues: spatial learning in the terrestrial toad Rhinella arenarum.

Although of crucial importance in vertebrate evolution, amphibians are rarely considered in studies of comparative cognition. Using water as reward, we studied whether the terrestrial toad, Rhinella arenarum, is also capable of encoding geometric and feature information to navigate to a goal location. Experimental toads, partially dehydrated, were trained in either a white rectangular box (Geometry-only, Experiment 1) or in the same box with a removable colored panel (Geometry-Feature, Experiment 2) covering one wall. Four water containers were used, but only one (Geometry-Feature), or two in geometrically equivalent corners (Geometry-only), had water accessible to the trained animals. After learning to successfully locate the water reward, probe trials were carried out by changing the shape of the arena or the location of the feature cue. Probe tests revealed that, under the experimental conditions used, toads can use both geometry and feature to locate a goal location, but geometry is more potent as a navigational cue. The results generally agree with findings from other vertebrates and support the idea that at the behavioral-level geometric orientation is a conserved feature shared by all vertebrates.

Vulnerability of long-term memory to temporal delays in amphibians.

Two experiments with toads were designed to test the memory-decay hypothesis that extinction (i.e., nonreinforced) performance is a function of time since the last reinforcement. In Experiment 1, toads (Rhinella arenarum, formerly Bufo arenarum) received 15 daily acquisition trials each reinforced with access to water during 300 s in a runway and were then randomly assigned to one of 6 retention intervals (RIs): 1, 4, 8, 16, 32, and 64 days. Extinction started after the RI and lasted 8 additional daily trials. Overall extinction performance was a logarithmic function of the RI. Although 4 extinction trials produced similar performance than 4 days of RI (consistent with memory decay), 8 extinction trials produced lower performance than 8 days of RI (consistent with a decremental effect of nonreinforcement). In Experiment 2, two groups of toads received 15 daily acquisition trials each reinforced with access to water for either 30 or 600 s, thus producing two reward magnitudes. After an 8-day RI, extinction performance was weaker after training with the small, than with the large reward magnitude. These results suggest that, at least in early extinction, the instrumental performance of toads is strongly influenced by the time since the last reinforcement.