Nonreinforced Probe Trials Research Articles

The use of an intermittent schedule of reinforcement to evaluate detection dogs' generalization from smokeless-powder.

Odor generalization is essential for detection dogs. Regardless of its importance, limited research is available on detection dog odor generalization. The objectives of this study were (1) evaluate the use of an intermittent schedule of reinforcement to assess generalization in dogs and (2) evaluate olfactory generalization from a single exemplar of smokeless powder (SP). Dogs (N = 5) were trained to detect SP in an automated olfactometer under an intermittent schedule of reinforcement where only 60% of correct responses were reinforced. After training, eight non-reinforced probe trials were inserted within a session. A total of 15 testing odors were evaluated across 15 consecutive sessions (one odor/session). Six of the testing odors were control and the remaining testing odors were objects indirectly exposed to SP, objects that contained or were directly exposed to SP, single-base SP and diphenylamine (the main volatile present in the headspace of SP). Dogs' response rate to all testing odors except for the cotton gauzes and t-shirt cloths exposed to the headspace of SP, the simulated IED, and Getxent tubes exposed to direct contact with SP were statistically lower than their response rate of actual SP. The response rate to SP was not different across all 15 testing sessions suggesting that the intermittent schedule of reinforcement, maintained dog motivation and performance. Data show that the outlined method is a good approach to study generalization in detection dogs. These results also highlight dog generalization to SP varieties and associated odors.

The Alteration of Emotion Regulation Precedes the Deficits in Interval Timing in the BACHD Rat Model for Huntington Disease.

Huntington disease (HD) is an autosomal dominantly inherited, progressive neurodegenerative disorder which is accompanied by executive dysfunctions and emotional alteration. The aim of the present study was to assess the impact of emotion/stress on on-going highly demanding cognitive tasks, i.e., temporal processing, as a function of age in BACHD rats (a “full length” model of HD). Middle-aged (4–6 months) and old (10–12 months) rats were first trained on a 2 vs. 8-s temporal discrimination task, and then exposed to a series of bisection tests under normal and stressful (10 mild unpredictable foot-shocks) conditions. The animals were then trained on a peak interval task, in which reinforced fixed-interval (FI) 30-s trials were randomly intermixed with non-reinforced probe trials. After training, the effect of stress upon time perception was again assessed. Sensitivity to foot-shocks was also assessed independently. The results show effects of both age and genotype, with largely greater effects in old BACHD animals. The older BACHD animals had impaired learning in both tasks, but reached equivalent levels of performance as WT animals at the end of training in the temporal discrimination task, while remaining impaired in the peak interval task. Whereas sensitivity to foot-shock did not differ between BACHD and WT rats, delivery of foot-shocks during the test sessions had a disruptive impact on temporal behavior in WT animals, an effect which increased with age. In contrast, BACHD rats, independent of age, did not show any significant disruption under stress. In conclusion, BACHD rats showed a disruption in temporal learning in late symptomatic animals. Age-related modification in stress-induced impairment of temporal control of behavior was also observed, an effect which was greatly reduced in BACHD animals, thus confirming previous results suggesting reduced emotional reactivity in HD animals. The results suggest a staggered onset in cognitive and emotional alterations in HD, with emotional alteration being the earliest, possibly related to different time courses of degeneration in cortico-striatal and amygdala circuits.

Characterizing Olfactory Binary Mixture Interactions in Fischer 344 Rats Using Behavioral Reaction Times

Response times provide essential subthreshold perceptual data that extend beyond accuracy alone. Behavioral reaction times (RTs) were used to characterize rats' ability to detect individual odorants in a series of complimentary binary odorant mixture ratios. We employed an automated, liquid-dilution olfactometer to train Fischer 344 rats (N = 8) on an odor identification task using nonreinforced probe trials. Binary mixture ratios composed of aliphatic odorants (citral and octanol) were arranged such that relative contributions of the 2 components varied systematically by a factor of 1% (v/v). Odorant concentrations for the target (S+), control (S-), and mixture (S+:S-) odorants were presented relative to threshold for each rat. Rats were initially trained to respond by licking at a spout to obtain liquid reward for either citral or octanol as the reinforced target (S+) odorant. After achieving 100% accuracy, rats were transferred to variable ratio (VR 2) reinforcement for correct responding. Nonreinforced probe trials (2 per block of 22 trials) were tested for each mixture ratio and recorded as either S+ (rats lick-responded in the presence of the mixture) or S- (rats refrained from licking), thereby indicating detection of the trained, S+ odorant. To determine the perceived salience for each ratio, RTs (latency from odorant onset to lick response) were recorded for each trial. Consistent with previous studies, RTs for both odorants were shortest (~150-200ms) when the probe trials consisted of a single, monomolecular component. Binary mixtures that contained as little as 1% of the S-, nontarget odorant, however, were sufficiently different perceptually to increase behavioral RTs (i.e., rats hesitated longer before responding); RTs changed systematically as a function of the binary ratio. Interestingly, the rate of RT change was dependent on which odorant served as the S+, suggesting an asymmetric interaction between the 2 odorants. The data demonstrate the value of behavioral RT as a sensitive measure of suprathreshold perceptual responding.

Spatio-temporal Binding: An Adaptation of the Peak Interval Procedure to Assess Spatio-temporal Learning in Rats

Time and space are commonly approached as two distinct dimensions, and rarely combined together in a single task. In this project, we designed a new version of the peak interval procedure in which we included a spatial component. The aim of this experiment was to investigate the learning of a spatio-temporal rule in animals. Adult male Sprague-Dawley rats were placed in front of a 5 nose-poke wall where a given hole could be visited after a precise time after a sound onset in a Fixed Interval (FI) schedule. The animals were presented two different contexts, differentiable by the house-light (A-red or B-green), tone frequency (1 or 11kHz), and spatio-temporal combination rule given by lighting a specific hole and associating it to a specific FI value (i.e., Hole 2-FI 15s vs. Hole 4-FI 60s). Non-reinforced probe trials of longer duration than the FI values (45s for FI15 and 120s for FI60) were pseudo-randomly presented to ascertain temporal peak interval functions. After 8 weeks of training, all the animals were able to differentiate the two contexts as shown by the number of nose-pokes in the correct hole peaking at the appropriate time, while virtually no activity was recorded in the other holes. In a single Pavlovian session of 8 trials, we changed the spatio- temporal rule for half of the animals: in the second context, the hole 1 was reinforced at 30seconds. Here we showed that this one session was sufficient for the animals to learn the new spatio-temporal association. Contrary to the control group, the experimental group exhibited a behavioral pattern associated to each hole indicative of a possible spatio-temporal binding. Interestingly, the behavior in the other control Context, in which the spatio-temporal rule had not been changed, was preserved in both groups. The experiment was followed for an extra 10 days after the Pavlovian Shift. At the end of the experiment, all the animals were perfectly and fully adapted to new spatio-temporal rule. In conclusion, the study suggests that rats are able to learn a spatio-temporal association in a single session, and that the two (space, time) dimensions may be learned together and not separately.

Numerical acuity of fish is improved in the presence of moving targets, but only in the subitizing range

There is controversy in comparative psychology about whether on the one hand non-symbolic number estimation of small (≤4) and large numbers involves a single mechanism (an approximate number system), or whether on the other hand enumeration of the numbers 1-4 is accomplished by a separate mechanism, an object tracking system. To date, support for the latter hypothesis has come only from the different ratio-dependency of performance seen in the two numerical ranges, a reading that has been criticized on several grounds. In humans, the two-system hypothesis is supported by evidence showing that manipulation of the physical properties of the stimuli (e.g., the motion of the items) has dissimilar effects on small- and large-number discrimination. In this research, we studied this effect on guppies. Initially, fish were trained to simultaneously discriminate two numerical contrasts having the same easy ratio (0.50): one in the small-number (2 vs. 4) range and one in the large-number (6 vs. 12) range. Half of the fish were presented with moving items; the other half were shown the same stimuli without motion. Fish were then subjected to non-reinforced probe trials in the presence of a more difficult ratio (0.75: 3 vs. 4 and 9 vs. 12). Under both static and moving conditions, the fish significantly discriminated 6 versus 12, but not 9 versus 12 items. As regards small numbers, both groups learned to discriminate a 0.50 ratio, but only fish tested with moving stimuli also discriminated 3 and 4 items. This differential effect suggests that fish may possess two separate systems for small- and large-number discrimination.

Relational responding by eastern box turtles (Terrapene carolina) in a series of color discrimination tasks.

Prior work with chelonians has demonstrated their capacity for successful performance in cognitive tasks, including those requiring color discrimination. Here, we sought to expand on historical research and determine whether eastern box turtles (Terrapene carolina) are capable of not only making simple color discriminations but also whether they can demonstrate abstract concept formation evidenced by using a relational response rule in their discrimination performance. Two eastern box turtles were rapidly and successfully trained on a black-and-white two-choice discrimination task using painted paddles and food reinforcement. After mastery, a medium gray paddle was added as a choice stimulus and turtle "Flippy" was reinforced for selecting the darker of the 2 stimuli presented in each trial, and turtle "Mario" was reinforced for selecting the lighter of the paddles presented. Nonreinforced probe trials incorporating light and dark gray stimuli paired with all other color options were then added to each session to test the turtles' ability to use the relationship between choice stimuli to guide responding. The turtles successfully selected the paddles corresponding to their assigned relational response rule of "darker" or "lighter" at a level significantly above that predicted by chance. The turtles then demonstrated immediate generalization of their relational rule in testing with a novel array of blue paddles. Finally, the turtles continued to use their relational rule when presented with a novel array of green paddles in a traditional transposition task. Together, these findings support the capacity for higher order cognitive functioning in chelonians beyond that previously described.

Inter-Specific Differences in Numerical Abilities Among Teleost Fish

Adults, infants and non-human primates are thought to possess similar non-verbal numerical systems, but there is considerable debate regarding whether all vertebrates share the same numerical abilities. Despite an abundance of studies, cross-species comparison remains difficult because the methodology employed and the context of species examination vary considerably across studies. To fill this gap, we used the same procedure, stimuli, and numerical contrasts to compare quantity abilities of five teleost fish: redtail splitfin, guppies, zebrafish, Siamese fighting fish, and angelfish. Subjects were trained to discriminate between two sets of geometrical figures using a food reward. Fish initially were trained on an easy numerical ratio (5 vs. 10 and 6 vs. 12). Once they reached the learning criterion, they were subjected to non-reinforced probe trials in which the set size was constant but numerical ratios varied (8 vs. 12 and 9 vs. 12). They also were subjected to probe trials in which the ratio was constant, but the total set size was increased (25 vs. 50) or decreased (2 vs. 4). Overall, fish generalized to numerosities with a 0.67 ratio, but failed with a 0.75 ratio; they generalized to a smaller set size, but not to a larger one. Only minor differences were observed among the five species. However, in one species, zebrafish, the proportion of individuals reaching the learning criterion was much smaller than in the others. In a control experiment, zebrafish showed a similar lower performance in shape discrimination, suggesting that the observed difference resulted from the zebrafish’s difficulty in learning this procedure rather than from a cross-species variation in the numerical domain.

Reflexivity in pigeons.

A recent theory of pigeons' equivalence-class formation (Urcuioli, 2008) predicts that reflexivity, an untrained ability to match a stimulus to itself, should be observed after training on two "mirror-image" symbolic successive matching tasks plus identity successive matching using some of the symbolic matching stimuli. One group of pigeons was trained in this fashion; a second group was trained similarly but with successive oddity (rather than identity). Subsequently, comparison-response rates on novel matching versus mismatching sequences with the remaining symbolic matching stimuli were measured on nonreinforced probe trials. Higher rates were observed on matching than on mismatching probes in the former group. The opposite effect--higher rates on mismatching than matching probes--was mostly absent in the latter group, despite being predicted by the theory. Nevertheless, the ostensible reflexivity effect observed in former group may be the first time this phenomenon has been demonstrated in any animal.

Averaging of temporal memories by rats.

Rats were trained on a mixed fixed-interval schedule in which stimulus A (tone or light) indicated food availability after 10 s and stimulus B (the other stimulus) indicated food availability after 20 s. Testing consisted of nonreinforced probe trials in which the stimulus was A, B, or the compound AB. On single-stimulus trials, rats responded with a peak of activity around the programmed reinforced time. On compound-stimulus trials, rats showed a single scalar peak of responding at a time midway between those for stimulus A and B. These results suggest that when provided with discrepant information regarding the temporal predictability of reinforcement, rats compute an average of the scheduled reinforcement times for the A and B stimuli and use this average to generate an expectation of reward for the compound stimuli.

Temporal generalization accounts for response resurgence in the peak procedure

The peak interval (PI) procedure is commonly used to evaluate animals' ability to produce timed intervals. It consists of presenting fixed interval (FI) schedules in which some of the trials are replaced by extended non-reinforced trials. Responding will often resume (resurge) at the end of the non-reinforced trials unless precautions are taken to prevent it. Response resurgence was replicated in rats and pigeons. Variation of the durations of the FI and the non-reinforced probe trials showed it to be dependent on the time when reinforcement is expected. Timing of both the normal time to reinforcement, and the subsequent time to reinforcement during the probe trials followed Weber's law. A quantitative model of resurgence is described, suggesting how animals respond to the signaling properties of reinforcement omission. Model results were simulated using a stochastic binary counter.

How do pigeons represent numbers?

Pigeons' bisection of two number scales, 1-16 and 2-32, was examined in two experiments. In Experiment 1, pigeons learned to choose different comparison keys following 1 or 16 key pecks and 2 or 32 key pecks and then were tested with intermediate numbers of key pecks on nonreinforced probe trials. Psychophysical curves that plotted choice of the large number key against number of pecks showed superposition of the two number scales and indifference between the two keys at the geometric means of each scale. The same procedures were used in Experiment 2, but pigeons were trained to bisect each scale at its arithmetic mean. Two asymmetries appeared in curves that plotted discrimination performance against number: near the midpoints of the scales, accuracy was higher for high numbers than for low numbers, but this relationship reversed at the ends of the scales, with low numbers discriminated more accurately than high numbers. An associative model that assumed stimulus generalization between numbers was used to predict the findings of these two experiments. The model showed a good qualitative fit to the obtained data when it was assumed that number was represented on a log scale and generalization was constant at all numbers but not when it was assumed that number was represented on a linear scale and generalization was scalar.

A new approach to the formation of equivalence classes in pigeons.

Four pigeons were given simultaneous discrimination training with visual patterns arbitrarily divided into two sets, with the stimuli in one set designated A1, B1, C1, and D1 and those in the other set designated A2, B2, C2, and D2. In sequentially introduced training phases, the pigeons were exposed to a series of reversals to establish AB and then CD equivalences. In subsequent testing sessions, a subset of stimuli from one set served as positive stimuli and those from the other set as negative stimuli on training trials, and transfer of the reinforced relation to other members of the sets was tested with nonreinforced probe trials. The pigeons were trained further on AC and BD equivalences and then were tested for the emergence of untrained AD and BC equivalences. Two of the 4 pigeons exhibited the emergence of one of these untrained equivalences, evidence for the emergence of transitive relations. This finding suggests that the pigeons established three-member functional equivalence classes by incorporating separately trained multiple equivalence relations. Repeated reversal training and probe testing enabled us to explore the formation and expansion of functional equivalence classes in pigeons.

Role of right hemifield in visual control of approach to target in zebrafish

When a zebrafish has to choose between two identical stimuli (e.g. a conditioned stimulus, CS, for food reward), it tends to respond to the one on its right. Errors are more numerous when reinforced for taking the one on the left rather than the one on the right. When trained to a single medial stimulus, and presented in non-reinforced probe trials with a pair of identical stimuli, the one on the right is chosen. Use by zebrafish of right eye (RE), viewing to control a planned motor response, extends from objects that are to be bitten to a choice of one of two routes. When the CS is visible behind a barrier of vertical bars, so that it can be approached around either end, it is the right end that is chosen. Standing motor bias independent of the nature of the task can be excluded. Other vertebrates show RE control of response. Toads are more likely to take food seen with the RE. The domestic chick uses the RE in visual control of approach to an object that has to be manipulated with the bill. RE control of use of the mouth in a fish shows that that this is an earlier condition than lateralised control of bilateral effectors like hands.