Fruit flies learn to avoid odours associated with virulent infection

While learning to avoid toxic food is common in mammals and occurs in some insects, learning to avoid cues associated with infectious pathogens has received little attention. We demonstrate that Drosophila melanogaster show olfactory learning in response to infection with their virulent intestinal pathogen Pseudomonas entomophila. This pathogen was not aversive to taste when added to food. Nonetheless, flies exposed for 3 h to food laced with P. entomophila, and scented with an odorant, became subsequently less likely to choose this odorant than flies exposed to pathogen-laced food scented with another odorant. No such effect occurred after an otherwise identical treatment with an avirulent mutant of P. entomophila, indicating that the response is mediated by pathogen virulence. These results demonstrate that a virulent pathogen infection can act as an aversive unconditioned stimulus which flies can associate with food odours, and thus become less attracted to pathogen-contaminated food.     

Diet palatability and growth efficiency: Evidence for a physiological interrelationship in rats

The total food intake and growth efficiency of growing rats were not affected by the feeding of a mild protein restricted (10%) diet containing an aversive taste stimulus. However, growing rats fed the same diet for a period of 18 days, suffered an inhibition of growth efficiency if the taste character of the diet was changed daily by the addition of a single but different aversive stimulus. For a period of time (9 days), these changes in diet palatability did not affect the total food intake. Rats fed diets containing a combination of the aversive taste changes and commercial soybean trypsin inhibitors, suffered an additional inhibition of growth efficiency. It is postulated that these manipulations in diet palatability interfered with digestive or metabolic processes.     

60 Hz electric fields and incandescent light as aversive stimuli controlling the behavior of rats responding under concurrent schedules of reinforcement

Several reports have shown that animals will sometimes engage in behaviors that reduce their exposure to a 60 Hz electric field (E-field). The field, therefore, can function as an aversive stimulus. In other studies, the E-field at equivalent strengths failed to function as an aversive stimulus. The present experiment, using rats, demonstrates how factors other than field strength can influence whether a subject engages in behavior that reduces field exposure. The general design consisted of giving the rat a choice between two alternatives, one of which sometimes included an added stimulus. Each subject was trained to press each of two levers to obtain food. Pressing one lever was reinforced intermittently under a variable interval 2 min schedule (VI 2); pressing the other lever was reinforced by a second VI 2 schedule operating independently of the first. Under this concurrent schedule the rat spent 50% of the daily 50 min session responding to each of the levers, indicating that they were equally “valued.” Next, while the schedules remained in effect, the first response to one of the levers turned on a 100 kV/m E-field which remained on until the rat pressed the other lever. The time spent responding under the schedule associated with the field was reduced by about 5–10%. When the procedure was changed so that no lever presses produced food, i.e., extinction, but the added stimulus contingency remained, the rats spent even less time in the presence of the field. Similar outcomes were observed during both the concurrent food or extinction schedules when incandescent light was used. Thus, both an E-field and incandescent light functioned as aversive stimuli, but the magnitude of the aversiveness was small. Aversiveness depended not only on stimulus intensity, but also on behavioral factors. Bioelectromagnetics 19:210–221, 1998. © 1998 Wiley-Liss, Inc.     

Sex differences in the cerebral lateralization of a cichlid fish when detouring to view emotionally conditioned stimuli

The lateralization of emotion has been described in a variety of animals. The right hemisphere has been implicated in the processing of negative emotions while positive emotions are processed in the left. Most animal studies of this phenomenon to date have used intrinsically emotionally arousing stimuli and there are few examples of lateralized responses to learned emotional triggers. It is known that males and females may demonstrate different patterns of lateralization, and that these sex differences may interact with other variables. We investigated the lateralized response of male and female convict cichlids to emotionally conditioned stimuli. One stimulus was given an appetitive (positive emotional valence) association by pairing with food. The other stimulus was given an aversive (negative emotional valence) association by pairing with a chemical alarm signal. We found that males tend to be more strongly lateralized to aversive stimuli while females are more strongly lateralized when responding to appetitive stimuli.     

Is avoiding an aversive outcome rewarding? Neural substrates of avoidance learning in the human brain

Avoidance learning poses a challenge for reinforcement-based theories of instrumental conditioning, because once an aversive outcome is successfully avoided an individual may no longer experience extrinsic reinforcement for their behavior. One possible account for this is to propose that avoiding an aversive outcome is in itself a reward, and thus avoidance behavior is positively reinforced on each trial when the aversive outcome is successfully avoided. In the present study we aimed to test this possibility by determining whether avoidance of an aversive outcome recruits the same neural circuitry as that elicited by a reward itself. We scanned 16 human participants with functional MRI while they performed an instrumental choice task, in which on each trial they chose from one of two actions in order to either win money or else avoid losing money. Neural activity in a region previously implicated in encoding stimulus reward value, the medial orbitofrontal cortex, was found to increase, not only following receipt of reward, but also following successful avoidance of an aversive outcome. This neural signal may itself act as an intrinsic reward, thereby serving to reinforce actions during instrumental avoidance.     

Brain regions responsible for the expression of conditioned taste aversion in rats

Conditioned taste aversion (CTA) is acquired when the ingestion of a food is followed by malaise. CTA is a kind of fear learning making animals avoid subsequent intake of the food and show aversive behavior to the taste of the food. To elucidate the brain regions responsible for the expression of CTA, our previous electrophysiological and recent c-fos immunohistochemical studies have been reviewed. Among a variety of brain regions including the parabrachial nucleus, amygdala, insular cortex, supramammillary nucleus, nucleus accumbens, and ventral pallidum that are involved in different phases of CTA expression, the enhanced taste sensitivity to facilitate detection of the conditioned stimulus may originate in the central nucleus of the amygdala and the hedonic shift, from positive to negative, may originate in the basolateral nucleus of the amygdala.     

Met-enkephalin effects on associations formed in utero

Rat fetuses exposed to an odor stimulus and an aversive stimulus in utero showed an aversion to the odor when tested 16 days postnatally. Fetuses that also received 80 μg/kg Met-enkephalin showed a greater aversion to the odor stimulus than those subjects that did not receive the peptide. The difference between these groups was marginally significant. Control subjects did not show an aversion. But, subjects exposed to the odor and Met-enkephalin without the aversive stimulus, when tested, showed a significant preference for the odor over other control groups. These data show that associative learning in rat fetuses at 20 days of gestation may be enhanced by administration of Met-enkephalin.     

Glutamate release in the nucleus accumbens during competitive presentation of aversive and appetitive stimuli

By means of in vivo microdialysis combined with HPLC analysis, we have shown that extracellular glutamate level in the rat n. accumbens increases during a simultaneous presentation of a palatable diet and a tone previously paired with a footshock, the magnitude of the extracellular glutamate increase being proportional to the latency of food taking. In contrast, extracellular glutamate level remains unchanged when the diet is presented after the conditioned aversive stimulus or when the tone is given alone. These data suggest that the glutamate release evoked by the competitive presentation of the diet and the conditioned aversive stimulus appears to be related to the inhibition of a planned feeding response, whereas the choice between behavioural strategies may not contribute to this phenomenon.     

Dairy cows' fear of people: social learning, milk yield and behaviour at milking

We examined the effects of the presence of an unfamiliar, a gentle or an aversive handler during milking on behaviour and milk yield, and whether cows can learn to approach or avoid a handler by observing the neighbouring cow's responses. In Experiment 1, Danish Friesian cows (n=16) were treated gently (offering hay and concentrates) by one handler and aversively (hit every 15s on the head with the hand) by another handler for six periods of 2min each. The two handlers wore different coloured overalls, and each cow received either gentle or aversive treatment in the first week and the other treatment the following week. All cows kept a longer distance to the aversive than to the gentle handler in a 1min test after treatment. Milk yield and residual milk did not differ when the aversive or the gentle handler was standing in front of the cow during milking, although the cows moved their legs and tail less when the aversive handler was present. When an unfamiliar person was standing in front of the cows during milking, behaviour and milk yield did not differ from control milkings. Cows and heifers (n=10) that had observed their neighbours receiving gentle treatment by one handler and aversive treatment from another handler did not differ in the distance they kept from these two handlers. In Experiment 2, cows (n=15) that had observed the neighbours receiving a gentle treatment (eight times for 2min) kept a shorter distance to that handler after treatment of their neighbours, and the distance they kept was correlated with the distance kept by the neighbouring cows. This suggests that responses of observer cows may be affected by the responses of the cows being treated. The cows rapidly learned to avoid an aversive handler, but although the cows showed clear avoidance response to the aversive handler there was no effect on milk yield when the aversive handler was present at milking.     

Comparison of 60-Hz electric fields and incandescent light as aversive stimuli controlling the behavior of rats

Rats were exposed to two procedures which enabled them to press a lever to turn off a 90 or 100 kV/m 60-Hz electric field or, later in the study, illumination from an incandescent lamp. Under one procedure, a response turned off the stimulus for a fixed duration, after which the stimulus was turned on again. A response during the off-period restarted the fixed duration. None of the rats turned the field off reliably. Next, under an alternative procedure, pressing one lever turned the field off; pressing the other lever turned it back on; responding under those conditions differed little from that seen at 0 kV/m. Under both procedures, when illumination from an incandescent lamp served as the stimulus, each rat did turn the stimulus off, and performances varied with stimulus intensity. The results show that a 100 kV/m 60-Hz electric field is not sufficient to function as an aversive stimulus under two procedures where illumination from a lamp does function as an aversive stimulus.     

Perception of loudness is influenced by emotion

Loudness perception is thought to be a modular system that is unaffected by other brain systems. We tested the hypothesis that loudness perception can be influenced by negative affect using a conditioning paradigm, where some auditory stimuli were paired with aversive experiences while others were not. We found that the same auditory stimulus was reported as being louder, more negative and fear-inducing when it was conditioned with an aversive experience, compared to when it was used as a control stimulus. This result provides support for an important role of emotion in auditory perception.     

Leg position learning by an insect. I. A heat avoidance learning paradigm

A new learning paradigm which employs a natural aversive stimulus, tightly constrains learned behavior and demonstrates learning in individual animals is described. Locusts were restrained so that only the femoro-tibial joint of a single metathoracic leg was free to move. Animals were required to maintain a particular range of femoro-tibial joint angle to avoid heating of the head. The position of the tibia was sampled by an on-line computer which set the limits of joint angle and controlled the aversive stimulus while storing data for further analysis. Control animals received patterns of aversive stimuli identical to those received by experimental animals but independent of their own joint position. Quantitative evaluation of the learned behavior of individual animals allowed the identification of three different behavioral strategies by which learning was achieved.     

Direct activation of the ventral striatum in anticipation of aversive stimuli

The brain "reward" system, centered on the limbic ventral striatum, plays a critical role in the response to pleasure and pain. The ventral striatum is activated in animal and human studies during anticipation of appetitive/pleasurable events, but its role in aversive/painful events is less clear. Here we present data from three human fMRI studies based on aversive conditioning using unpleasant cutaneous electrical stimulation and show that the ventral striatum is reliably activated. This activation is observed during anticipation and is not a consequence of relief after the aversive event. Further, the ventral striatum is activated in anticipation regardless of whether there is an opportunity to avoid the aversive stimulus or not. Our data suggest that the ventral striatum, a crucial element of the brain "reward" system, is directly activated in anticipation of aversive stimuli.     

Ontogenetic differences in sensitivity to LiCl- and amphetamine-induced taste avoidance in preweanling rats

When amphetamine is associated with a tastant conditioned stimulus, rats learn to avoid the taste even when employing doses that promote conditioned place preference. One hypothesis raised to account for this effect proposes that taste avoidance induced by amphetamine may be motivated by fear. A sensitive period has been identified in the rat (until postnatal day 10) in which infants learn conditioned appetitive effects to stimuli to which aversions are conditioned after this period. Exogenous administration of corticosterone within this period reverses this effect, generating aversive conditioning. In the present study, we tested conditioning of aversions to amphetamine or LiCl, within and after the sensitive period (Experiments 1 and 2). A third experiment evaluated unconditioned rejection of an aversive quinine solution within the sensitive period. Finally, we tested whether corticosterone administration before conditioning modulates amphetamine-induced taste avoidance. After the sensitive period, infant rats rejected the solution paired with amphetamine or LiCl after 2 conditioning trials, but within the sensitive period, aversions were conditioned only by LiCl and after 4 conditioning trials. Amphetamine-induced taste avoidance was not observed even when corticosterone was administered before conditioning. Additionally, during the sensitive period, a low LiCl dose promoted conditioned taste preference. According to Experiment 3, parameters employed in this study were suitable to yield rejection of aversive solutions within the sensitive period. These results suggest that during the sensitive period, there is a notable resistance to the acquisition of taste avoidance induced by amphetamine. The present experimental framework may represent a useful tool for studying mechanisms underlying taste avoidance and aversion effects.     

Comparison of central administration of corticotropin-releasing hormone and urocortin on food intake, conditioned taste aversion, and c-Fos expression

Corticotropin-releasing hormone (CRH) is a potent regulator of the hypothalamic-pituitary-adrenal axis, and reduces food intake when administered into the third cerebral ventricle (i3vt). However, CRH also promotes conditioned taste aversion (CTA) learning which indicates that its anorectic effects are accompanied by aversive consequences that would reduce food intake independently of energy regulation. Urocortin (Ucn) is a closely related mammalian peptide that binds to both identified CRH receptor subtypes and also reduces food intake when administered i3vt. The present experiments compared the aversive consequences of i3vt administration of CRH and Ucn at doses that produced comparable decrements in food intake. Experiment 1 found that 1.0 microg Ucn and 2.0 microg CRH produced similar reductions in food intake. Experiment 2 demonstrated that, at these doses, CRH but not Ucn promoted robust and reliable CTA learning. A third experiment showed comparable increased c-Fos-like immunoreactivity after Ucn and CRH in forebrain and hindbrain structures associated with food intake. It is concluded that Ucn, at doses that reduce food intake to levels like that observed after administration of CRH, do not produce similarly aversive consequences.     

Investigating Fear in Rainbow Trout (Oncorhynchus mykiss) Using the Conditioned-Suppression Paradigm

Trout learned the operant task of pendulum-pressing for a food-reward in a mean of 4.3 sessions lasting 1 hr. In a separate phase, fish also learned—through classical conditioning—to associate a neutral light cue with an aversive stimulus. When again allowed to pendulum-press for food, after aversive classical conditioning, there was a drop in the rate of responding. The mean rate dropped from 3.6–2.9 responses per min. Most important, when the light-stimulus was superimposed on a steady bout of pendulum-pressing, trout ceased to press the pendulum and did not resume activity until termination of the light-stimulus (mean number of responses during a 3-min interval immediately prior to light-stimulus = 14.3 vs. during 3-min light-stimulus = 0.1). Psychologists have used this decrease in operant responding, or “conditioned emotional response,” as a tool to examine the psychological nature of this type of aversive conditioning. In this study, the fish demonstrated various results under this paradigm similar to those shown by “higher” nonhuman animals, therefore challenging the view of fish as unconscious, nonsentient animals.     

Investigating fear in rainbow trout (Oncorhynchus mykiss) using the conditioned-suppression paradigm

Trout learned the operant task of pendulum-pressing for a food-reward in a mean of 4.3 sessions lasting 1 hr. In a separate phase, fish also learned—through classical conditioning—to associate a neutral light cue with an aversive stimulus. When again allowed to pendulum-press for food, after aversive classical conditioning, there was a drop in the rate of responding. The mean rate dropped from 3.6-2.9 responses per min. Most important, when the light-stimulus was superimposed on a steady bout of pendulum-pressing, trout ceased to press the pendulum and did not resume activity until termination of the light-stimulus (mean number of responses during a 3-min interval immediately prior to light-stimulus = 14.3 vs. during 3-min light-stimulus = 0.1). Psychologists have used this decrease in operant responding, or “conditioned emotional response,” as a tool to examine the psychological nature of this type of aversive conditioning. In this study, the fish demonstrated various results under this paradigm similar to those shown by “higher” nonhuman animals, therefore challenging the view of fish as unconscious, nonsentient animals.     

Appetitive and aversive learning in Spodoptera littoralis larvae

Adult Lepidoptera are capable of associative learning. This helps them to forage flowers or to find suitable oviposition sites. Larval learning has never been seriously considered because they have limited foraging capabilities and usually depend on adults as concerns their food choices. We tested if Spodoptera littoralis larvae can learn to associate an odor with a tastant using a new classical conditioning paradigm. Groups of larvae were exposed to an unconditioned stimulus (US: fructose or quinine mixed with agar) paired with a conditioned stimulus (CS: hexanol, geraniol or pentyl acetate) in a petri dish. Their reaction to CS was subsequently tested in a petri dish at different time intervals after conditioning. Trained larvae showed a significant preference or avoidance to CS when paired with US depending on the reinforcer used. The training was more efficient when larvae were given a choice between an area where CS-US was paired and an area with no CS (or another odor). In these conditions, the memory formed could be recalled at least 24 h after pairing with an aversive stimulus and only 5 min after pairing with an appetitive stimulus. This learning was specific to CS because trained larvae were able to discriminate CS from another odor that was present during the training but unrewarded. These results suggest that Lepidoptera larvae exhibit more behavioral plasticity than previously appreciated.     

桔小实蝇雄成虫联系性嗅觉学习

【目的】为了探究桔小实蝇 Bactrocera dorsalis (Hendel)雄成虫的嗅觉学习能力。【方法】本研究采用经典性嗅觉条件反射训练法（classical olfactory conditioning）在室内对固定的羽化后14-17日龄的桔小实蝇雄成虫进行气味与食物的联合学习训练, 即薄荷精油和10%蔗糖溶液联合的奖赏性训练（appetitive conditioning）以及甲基丁香酚(methyl eugenol, ME)和饱和盐溶液联合的惩罚性训练（aversive conditioning）,并以伸喙反射行为（proboscis extension reflex, PER）作为学习与否的判定标准。【结果】经过奖赏性训练后,桔小实蝇雄成虫对薄荷精油的伸喙反射率可从0%增加至68%;而经过惩罚性训练后,桔小实蝇对甲基丁香酚的伸喙反射率可从100%降低至36.54%,且这种伸喙反射率的变化是通过气味条件刺激（conditioned stimulus）和食物非条件刺激（unconditioned stimulus）的对称性联合而产生的。【结论】结果表明,桔小实蝇雄性成虫具有较强的联系性嗅觉学习能力,并且两种刺激的联合是形成学习记忆的必要条件。