Neutral Odor Research Articles

Anxiolytic-like effect of lavender essential oil inhalation in mice: Participation of serotonergic but not GABAA/benzodiazepine neurotransmission

Ethnopharmacological relevanceLavandula angustifolia (lavender) inhalation has been used in folk medicine for the treatment of anxiety, and clinical and animal studies have corroborated its anxiolytic effect, although its mechanism of action is still not fully understood. Aims of the studyThe objective of the present study was to determine whether the GABAA/benzodiazepine complex or serotonin neurotransmission mediates the anxiolytic-like effect of lavender essential oil. Materials and methodsMale Swiss mice were subjected to the marble-burying test after being exposed to the aroma of lavender essential oil (1–5%), amyl acetate (5%; used as a behaviorally neutral odor), or distilled water for 15min via inhalation. Additionally, the effect of 5% lavender essential oil was also evaluated in mice subjected to the elevated plus maze. GABAA/benzodiazepine mediation was evaluated by pretreating the mice with the GABAA receptor antagonist picrotoxin before the marble burying test and [3H]flunitrazepam binding to the benzodiazepine site on the GABAA receptor. Serotonergic mediation was studied by pretreating the mice with O-methyl-[3H]-N-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-N-(2-pyridinyl) cyclohexanecarboxamide trihydrochloride (WAY100635), a serotonin 5-HT1A receptor antagonist before the marble burying test. We also evaluated changes in the pharmacologically induced serotonin syndrome and the effects of combined administration of subeffective doses of lavender essential oil and the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). ResultsLavender essential oil (1–5%) decreased the number of marbles buried compared with the control and amyl acetate groups. In the elevated plus maze, 5% lavender essential oil inhalation increased the percentage of time spent on and number of entries into the open arms compared with controls. No effect was seen in the number of closed arm entries or number of beam interruptions in the automated activity chamber. Pretreatment with the GABAA receptor antagonist picrotoxin (0.5mg/kg) did not modify the behavioral effect of 5% lavender essential oil in the marble-burying test. Lavender essential oil also did not alter [3H]flunitrazepam binding to the benzodiazepine site on the GABAA receptor. Pretreatment with the serotonin 5-HT1A receptor antagonist WAY100635 (3mg/kg) blocked the anxiolytic-like effect of lavender essential oil and the 5-HT1A receptor agonist 8-OH-DPAT (3mg/kg). A combination of ineffective doses of 8-OH-DPAT (0.5mg/kg) and lavender essential oil (0.1%) reduced the number of marbles buried. Finally, 5% lavender essential oil attenuated the serotonin syndrome induced by 40mg/kg fluoxetine plus 80mg/kg 5-hydroxytryptophan. ConclusionsThese results indicate an important role for the serotonergic system in the anxiolytic-like effect of lavender essential oil.

Differential ultrasonic indices of separation distress in the presence and absence of maternal cues in infant rats bred for high and low positive social affect.

Preclinical models of human mood disorders commonly focus on the study of negative affectivity, without comparably stressing the role of positive affects and their ability to promote resilient coping styles. We evaluated the role of background constitutional affect of rats by studying the separation and reunion responses of infants from low and high positive affect genetic lines (i.e., differentially selected for High and Low 50 kHz ultrasonic vocalisations (USVs). Infants from Low and High 50 kHz USV breeding lines were isolated from mothers and exposed to either social (familiar or unfamiliar bedding) or neutral (clean bedding) odour cues between two short isolation periods, and tested in homeothermic and hypothermic ambient temperatures. Negative affect was estimated by monitoring separation distress calls (35-45 kHz USVs). Low Line pups called at higher rates than High Line, and their rates were stable regardless of odour cue. In contrast, High Line pups increased vocalisations during the second compared with the first isolation periods and during exposure to both familiar and unfamiliar odour cues, but not to neutral odour. Furthermore, the greatest increase in USV emission was seen in the second isolation period following exposure to the unfamiliar odour. However, both lines showed comparable elevated distress USVs to the thermal stressor. High Line animals, selected for a positive affective phenotype (50 kHz USVs), exhibited reduced separation anxiety responses in infancy, making this a promising animal model for the role of constitutional affective states in emotional responsivity and potential resilience against emotional disorders.

Candidate odours for a short form odorant identification test

Measurement of olfactory function is difficult and despite the availability of validated olfactory tests these are not in common use in general clinical practice. The mismatch between self-reported olfactory function and objective test response is well known, thus the need for adequate assessment of olfaction is not disputed. We aimed to determine which if any odour in a commercially available olfactory test could be identified by 70 % or more of normal subjects with a view to developing a shortened screening test for use in general otolaryngology and non-specialist clinics in the United Kingdom. Hospital staff and medical students, under 50 years of age were recruited. The tests were undertaken in a quiet room of neutral odour. Subjects were asked to self-identify 12 odorants presented consecutively and to record how strongly they perceived the odorants on a 4-point scale. One hundred and four participants correctly identified the odorant in 466 (37.3 %) of the 1,248 individual odorant identification tests. Peppermint, fish and coffee odours were identified by 88.5, 79.8 and 69.7 % of participants, respectively, without the advantage of a choice of name prompt. These supra-threshold test odour stimuli were perceived as present in all of the individual odour identification tests, but correct identification of the test odour was very much lower for the other nine test stimuli. Peppermint, fish and coffee are odours that should be evaluated further for inclusion in a short form olfactory identification test in a British population.

Men’s Exposure to Women’s Odors: The Effect of Women’s Waist to Hip Ratio and Steroid Hormones

Previous research has shown that axillar and vaginal odors from ovulating women are recognized by men, but no research has been done exploring whether body odors signal other physical attributes of women such as waist-to-hip ratio (WHR) or endocrine status. Our goal was to investigate whether testosterone increased in men after smelling axillary odors of women with different WHR and with low or high levels of steroid hormones. We measured men’s testosterone before and after 30 min of being exposed to fresh axillar odors collected from young women or a neutral odor as control situation. Men had to rate the attractiveness and intensity of the scent. We compared the response of men according to women’s WHR and salivary testosterone, estradiol and progesterone. Although the main literature commonly reports that men judge a WHR around 0.7 to be more visually attractive, our results showed that men rated samples of high WHR (0.75-0.84) and high estradiol women as more attractive. In addition, men’s testosterone increased after smelling the odors of high WHR, high estradiol and high testosterone women. High WHR women exhibited the highest testosterone and estradiol levels compared to the other WHR categories (0.66-0.74). We concluded that scents are cues that not only signal fertility but also physical attributes related to reproductive health.

It's time to fear! Interval timing in odor fear conditioning in rats

Time perception is crucial to goal attainment in humans and other animals, and interval timing also guides fundamental animal behaviors. Accumulating evidence has made it clear that in associative learning, temporal relations between events are encoded, and a few studies suggest this temporal learning occurs very rapidly. Most of these studies, however, have used methodologies that do not permit investigating the emergence of this temporal learning. In the present study we monitored respiration, ultrasonic vocalization (USV) and freezing behavior in rats in order to perform fine-grain analysis of fear responses during odor fear conditioning. In this paradigm an initially neutral odor (the conditioned stimulus, CS) predicted the arrival of an aversive unconditioned stimulus (US, footshock) at a fixed 20-s time interval. We first investigated the development of a temporal pattern of responding related to CS-US interval duration. The data showed that during acquisition with odor-shock pairings, a temporal response pattern of respiration rate was observed. Changing the CS-US interval duration from 20-s to 30-s resulted in a shift of the temporal response pattern appropriate to the new duration thus demonstrating that the pattern reflected the learning of the CS-US interval. A temporal pattern was also observed during a retention test 24 h later for both respiration and freezing measures, suggesting that the animals had stored the interval duration in long-term memory. We then investigated the role of intra-amygdalar dopaminergic transmission in interval timing. For this purpose, the D1 dopaminergic receptors antagonist SCH23390 was infused in the basolateral amygdala before conditioning. This resulted in an alteration of timing behavior, as reflected in differential temporal patterns between groups observed in a 24 h retention test off drug. The present data suggest that D1 receptor dopaminergic transmission within the amygdala is involved in temporal processing.

Neural processing of a cocaine-associated odor cue revealed by functional MRI in awake rats

Using an olfactory conditioning procedure, brain stimulation reward threshold measurements, and functional magnetic resonance imaging (fMRI), we investigated brain stimulation reward threshold change and fMRI neural activation in response to a cocaine-associated odor cue. In the first brain stimulation experiment, over 10 days of rate–frequency curve-shift testing, rats were administered intravenous cocaine (1.0mg/kg) paired with a contextual cue of peppermint odor previously placed in the operant chamber or they were given vehicle treatment (no cocaine) in the presence of no olfactory cue. Following a 14-day drug-free rest period, rats were again given the rate–frequency curve-shift threshold test with or without the odor cue. In a second experiment, rats were similarly conditioned with a peppermint odor but with intraperitoneally delivered cocaine (10mg/kg). After a 14 day rest period, rats were imaged on a 7-T MRI for their blood oxygen level dependent (BOLD) in response to the cocaine-paired peppermint odor versus an unpaired neutral lemon odor. In the brain stimulation experiment, expected significant reward threshold shifts were produced by cocaine and, importantly, about half that level of shift was produced by the paired contextual olfactory cue. In the fMRI experiment, the insular cortex showed a significantly greater BOLD activation in cocaine-treated versus saline-treated animals to the olfactory cue, but not with the unpaired lemon scent. These data are in agreement with previous studies suggesting a role of the insular cortex in attributing reward value (positive or negative) to conditioned odor stimuli.

Visualization of odor-induced neuronal activity by immediate early gene expression

BackgroundSensitive detection of sensory-evoked neuronal activation is a key to mechanistic understanding of brain functions. Since immediate early genes (IEGs) are readily induced in the brain by environmental changes, tracing IEG expression provides a convenient tool to identify brain activity. In this study we used in situ hybridization to detect odor-evoked induction of ten IEGs in the mouse olfactory system. We then analyzed IEG induction in the cyclic nucleotide-gated channel subunit A2 (Cnga2)-null mice to visualize residual neuronal activity following odorant exposure since CNGA2 is a key component of the olfactory signal transduction pathway in the main olfactory system.ResultsWe observed rapid induction of as many as ten IEGs in the mouse olfactory bulb (OB) after olfactory stimulation by a non-biological odorant amyl acetate. A robust increase in expression of several IEGs like c-fos and Egr1 was evident in the glomerular layer, the mitral/tufted cell layer and the granule cell layer. Additionally, the neuronal IEG Npas4 showed steep induction from a very low basal expression level predominantly in the granule cell layer. In Cnga2-null mice, which are usually anosmic and sexually unresponsive, glomerular activation was insignificant in response to either ambient odorants or female stimuli. However, a subtle induction of c-fos took place in the OB of a few Cnga2-mutants which exhibited sexual arousal. Interestingly, very strong glomerular activation was observed in the OB of Cnga2-null male mice after stimulation with either the neutral odor amyl acetate or the predator odor 2, 3, 5-trimethyl-3-thiazoline (TMT).ConclusionsThis study shows for the first time that in vivo olfactory stimulation can robustly induce the neuronal IEG Npas4 in the mouse OB and confirms the odor-evoked induction of a number of IEGs. As shown in previous studies, our results indicate that a CNGA2-independent signaling pathway(s) may activate the olfactory circuit in Cnga2-null mice and that neuronal activation which correlates to behavioral difference in individual mice is detectable by in situ hybridization of IEGs. Thus, the in situ hybridization probe set we established for IEG tracing can be very useful to visualize neuronal activity at the cellular level.

Pheromone-induced odor learning modifies Fos expression in the newborn rabbit brain

Associative learning contributes crucially to adjust the behavior of neonates to the permanently changing environment. In the European rabbit, the mammary pheromone (MP) excreted in milk triggers sucking behavior in newborns, and additionally promotes very rapid learning of initially neutral odor cues. Such stimuli become then as active as the MP itself to elicit the orocephalic motor responses involved in suckling. In this context, the rabbit is an interesting model to address the question of brain circuits early engaged by learning and memory. Here, we evaluated the brain activation (olfactory bulb and central regions) induced in 4-day-old pups by an odorant (ethyl acetoacetate, EAA) after single pairing with the MP and its subsequent acquired ability to elicit sucking-related behavior (conditioned group) or after mere exposure to EAA alone (unconditioned group). The brain-wide mapping of c-Fos expression was used to compare neural activation patterns in both groups. Evidence of high immunostaining to odorant EAA occurred in the mitral+granule cells layer of the main olfactory bulb in pups previously exposed to EAA in association with the MP. These pups also showed higher expression of Fos in the piriform cortex, the hypothalamic lateral preoptic area and the amygdala (cortical and basal nuclei). Thus, MP-induced odor learning induces rapid brain modifications in rabbit neonates. The cerebral framework supporting the acquisition appears however different compared to the circuit involved in the processing of the MP itself.

Behavioral responses of native prey to disparate predators: naiveté and predator recognition

It is widely accepted that predator recognition and avoidance are important behaviors in allowing prey to mitigate the impacts of their predators. However, while prey species generally develop anti-predator behaviors through coevolution with predators, they sometimes show accelerated adoption of these behaviors under strong selection pressure from novel species. We used a field manipulation experiment to gauge the ability of the common ringtail possum (Pseudocheirus peregrinus), a semi-arboreal Australian marsupial, to recognize and respond to olfactory cues of different predator archetypes. We predicted that ringtails would display stronger anti-predator behaviors to cues of the invasive European red fox (Vulpes vulpes) in areas where fox impacts had been greatest, and to cues of the native lace monitor (Varanus varius) in areas of sympatry compared with allopatry. We found that ringtails fled quickly and were more alert when exposed to the fecal odors of both predators compared to neutral and pungent control odors, confirming that predator odors are recognized and avoided. However, these aversive responses were similar irrespective of predator presence or level of impact. These results suggest that selection pressure from the fox has been sufficient for ringtails to develop anti-predator behaviors over the few generations since foxes have become established. In contrast, we speculate that aversive responses by ringtails to the lace monitor in areas where this predator is absent reflect recent coexistence of the two species. We conclude that rapid evolution of anti-predator behaviors may occur when selection is strong. The maintenance of these behaviors should allow re-establishment of predator-prey relationships if the interactants regain sympatry via range shifts or management actions to reintroduce them to their former ranges.

Olfactory stimulation curbs food cravings

Based on the logic of mutual competition between cravings and odours for limited-capacity resources, this study investigated whether a simple olfactory task, involving a brief odour exposure, could reduce food cravings. In support, Experiment 1 showed that smelling a neutral unfamiliar odorant reduced cravings for highly desired food items, relative to a comparison auditory task and a no-task control condition. Experiment 2 replicated these findings specifically for chocolate cravings, which can be particularly problematic. Thus olfactory stimulation offers potential scope for curbing unwanted food cravings.

Peripheral and Central Olfactory Tuning in a Moth

Animals can be innately attracted to certain odorants. Because these attractants are particularly salient, they might be expected to induce relatively strong responses throughout the olfactory pathway, helping animals detect the most relevant odors but limiting flexibility to respond to other odors. Alternatively, specific neural wiring might link innately preferred odors to appropriate behaviors without a need for intensity biases. How nonpheromonal attractants are processed by the general olfactory system remains largely unknown. In the moth Manduca sexta, we studied this with a set of innately preferred host plant odors and other, neutral odors. Electroantennogram recordings showed that, as a population, olfactory receptor neurons (ORNs) did not respond with greater intensity to host plant odors, and further local field potential recordings showed that no specific amplification of signals induced by host plant odors occurred between the first olfactory center and the second. Moreover, when odorants were mutually diluted to elicit equally intense output from the ORNs, moths were able to learn to associate all tested odorants equally well with food reward. Together, these results suggest that, although nonpheromonal host plant odors activate broadly distributed responses, they may be linked to attractive behaviors mainly through specific wiring in the brain.

Enhanced Olfactory Sensory Perception of Threat in Anxiety: An Event-Related fMRI Study

The current conceptualization of threat processing in anxiety emphasizes emotional hyper-reactivity, which mediates various debilitating symptoms and derangements in anxiety disorders. Here, we investigated olfactory sensory perception of threat as an alternative causal mechanism of anxiety. Combining an event-related functional magnetic resonance imaging paradigm with an olfactory discrimination task, we examined how anxiety modulates basic perception of olfactory threats at behavioral and neural levels. In spite of subthreshold presentation of negative and neutral odors, a positive systematic association emerged between negative odor discrimination accuracy and anxiety levels. In parallel, the right olfactory primary (piriform) cortex indicated augmented response to subthreshold negative (vs. neutral) odors as a function of individual differences in anxiety. Using a psychophysiological interaction analysis, we further demonstrated amplified functional connectivity between the piriform cortex and emotion-related regions (amygdala and hippocampus) in response to negative odor, particularly in anxiety. Finally, anxiety also intensified skin conductance response to negative (vs. neutral) odor, indicative of potentiated emotional arousal to subliminal olfactory threat in anxiety. Together, these findings elucidate exaggerated processing of olfactory threat in anxiety across behavioral, autonomic physiological, and neural domains. Critically, our data emphasized anxiety-related hyper-sensitivity of the primary olfactory cortex and basic olfactory perception in response to threat, highlighting neurosensory mechanisms that may underlie the deleterious symptoms of anxiety.

Rapid and Highly Resolving: Affective Evaluation of Olfactorily Conditioned Faces

Evidence from hemodynamic and electrophysiological measures suggests that the processing of emotionally relevant information occurs in a spatially and temporally distributed affective network. ERP studies of emotional stimulus processing frequently report differential responses to emotional stimuli starting around 120 msec. However, the involvement of structures that seem to become activated at earlier latencies (i.e., amygdala and OFC) would allow for more rapid modulations, even in distant cortical areas. Consistent with this notion, recent ERP studies investigating associative learning have provided evidence for rapid modulations in sensory areas earlier than 120 msec, but these studies either used simple and/or very few stimuli. The present study used high-density whole-head magneto-encephalography to measure brain responses to a multitude of neutral facial stimuli, which were associated with an aversive or neutral odor. Significant emotional modulations were observed at intervals of 50-80 and 130-190 msec in frontal and occipito-temporal regions, respectively. In the absence of contingency awareness and with only two learning instances, a remarkable capacity for emotional learning is observed.

Learned host preference in a Chagas disease vector, Rhodnius prolixus

It has been largely assumed that the individual experience of disease vector insects may affect host choice and, as a consequence, have an important influence on parasite transmission. In particular, it is speculated that vector insects should be able to learn and remember the most and/or less defensive hosts, shifting their preference accordingly. Nevertheless, despite the invested efforts in testing the capacity to learn and remember information of blood-sucking insects, only little conclusive information has been obtained hitherto. Recently, the ability of Rhodnius prolixus to associate a behaviourally neutral odour to the perspective of either obtaining a blood-meal or being punished has been demonstrated, the same odour becoming attractant or repellent for the bugs, respectively, according to the individual previous experience. The present work represents a step forward in the study of the cognitive abilities of Chagas disease vectors and their influence on host choice. We tested whether or not bugs bias their choice for a host based on the association of its odour with a negative experience. Our results show that whereas naïve bugs presented no preference when confronted to the odour of two different hosts, bugs previously exposed to the contingency of the odour of one host and a mechanical perturbation mimicking defensive behaviour, biased their preference towards the other host. This constitutes the first evidence of olfactory conditioning to host odours in triatomine bugs, vectors of Chagas disease and one of the few available up to date on haematophagous insects. The epidemiological significance of this finding is discussed.

OPTIMIZATION OF ACTIVATION CONDITIONS OF LOCALLY AVAILABLE CLAYS FOR COTTONSEED OIL BLEACHING: CHARACTERIZATION AND PERFORMANCE EVALUATION

ABSTRACTCottonseed oil is consumed either in raw state or after refining, which enables it to acquire the characteristics such as neutral taste and odor, limpidity, light color, oxidation stability to oxidation, and for it to be used for frying. Refining comprises several strategies including bleaching, which involves the fixation of beta‐carotene, the pigment that is responsible for the red color of cottonseed oils, on a solid support generally known as bleaching earth. In this work, possible application of locally available clay (Ordu, Turkey) as bleaching earth was investigated for cottonseed oil refining. Raw clay was activated via various treatment methods including acid activation, cation treatment and thermal treatment. Structural changes caused by acid activation were characterized using Fourier transform infrared spectroscopy, differential thermal analysis, N2 adsorption (BET surface area), X‐ray fluorescence and scanning electron microscopy. Bleaching capacities of the treated clays were also determined spectrophotometrically. Bleaching efficiency was increased from 3 (raw clay) to 23% by H2SO4 activation and to 34% by Al3+ and heat treatment. However, commercial clay's efficiency was 32%. Tetrahedral destruction with proton attack was higher in HCl solution than H2SO4 solution, resulting in collapse in micropores. Bleaching efficiency increment using H2SO4 was higher than that of HCl.PRACTICAL APPLICATIONSIt is important to reduce color in oils of low quality by cheap and appropriate adsorbent instead of expensive commercial earths. Activationconditions differ from one clay to another. In this study, several activation methods were applied and possible mechanisms relating to bleaching efficiency improvement were also investigated.

Brain processing of the mammary pheromone in newborn rabbits

Chemosignals strongly contribute to social interactions in mammals, including mother–young relationships. In the European rabbit, a volatile compound emitted by lactating females in milk, the 2-methylbut-2-enal, has been isolated. Carrying the properties of a pheromone, in particular the spontaneous ability to release critical sucking-related movements in newborns, it has been called the mammary pheromone (MP). Lesion of the vomeronasal organ and preliminary 2-deoxyglucose data suggested that the MP could be processed by the main olfactory system. However, the neuronal substrate that sustains the MP-induced response of neonates remained unknown. Here, we evaluated Fos expression in 4-day-old-rabbits exposed to the MP (in comparison with control neonates exposed to non-relevant odorant, no odorant or unmanipulated pups) both at the level of the olfactory bulb and central brain regions. Evidence of high and widespread Fos immunoreactivity in the main olfactory bulb appear in MP pups while the accessory olfactory bulb exhibits a negligible staining. However, no obvious bulbar pattern of Fos expression is observed, when in contrast a certain pattern emerges with the neutral odorant. Compared to this latter, the MP exposure increases Fos expression in the anterior piriform cortex, the organum vasculosum of the lamina terminalis and the habenula, with a tendency in the lateral preoptic region. For the first time, a pheromone essential for mother–young interaction is thus highlighted for its processing by the main olfactory system, the whole olfactory bulb, and by brain regions involved in osmoregulation, thirst and motivation-guided motor responses.

Learning the way to blood: first evidence of dual olfactory conditioning in a blood-sucking insect, Rhodnius prolixus. II. Aversive learning

After having demonstrated that blood-sucking bugs are able to associate a behaviourally neutral odour (L-lactic acid) with positive reinforcement (i.e. appetitive conditioning) in the first part of this study, we tested whether these insects were also able to associate the same odour with a negative reinforcement (i.e. aversive conditioning). Learned aversion to host odours has been repeatedly suggested as a determinant for the distribution of disease vectors among host populations. Nevertheless, no experimental evidence has been obtained so far. Adapting a classical conditioning approach to our haematophagous model, we trained larvae of Rhodnius prolixus to associate L-lactic acid, an odour perceived by bugs but behaviourally neutral when presented alone, with a mechanical perturbation (i.e. negative reinforcement). Naive bugs and bugs exposed to CS, punishment, or CS and punishment without contingency remained indifferent to the presence of an air stream loaded with L-lactic acid (random orientation on a locomotion compensator), whereas the groups previously exposed to the contingency CS-punishment were significantly repelled by L-lactic acid. In a companion paper, the opposite, i.e. attraction, was induced in bugs exposed to the contingency of the same odour with a positive reinforcement. These constitute the first pieces of evidence of olfactory conditioning in triatomine bugs and the first demonstration that the same host odour can be used by insects that are disease vectors to learn to recognize either a host to feed on or a potentially defensive one. The orientation mechanism during repulsion is also discussed in light of our results.

Synergetic deodorant effect and antibacterial activity of composite paper containing waste tea leaves

Composite paper containing waste tea leaves was prepared to investigate the effective utilization of waste tea leaves as deodorant material. Paper containing waste green tea leaves did not have increased deodorizing ability compared with controls either against acidic odors such as hydrogen sulfide and acetic acid gases or against neutral odors such as formaldehyde and acetaldehyde gases. In contrast, the paper had excellent deodorizing ability against basic odors such as ammonia and trimethylamine gases. It was observed during additional tests conducted on paper samples containing 60 wt% waste leaves of oolong tea, black tea, pu-erh tea, or hojicha, that all the samples reduced the ammonia concentration to below 1 ppm, which is the threshold concentration for olfactory recognition, within 30 min. Further, paper containing waste green tea leaves was found to decrease the odor residual rate to 5.1% in 30 min even for a waste tea leaf content of 10 wt%. The excellent deodorizing ability of the paper could be attributed to the chemical reactions between odorous substances and the catechins found in tea leaves. After the deodorization of ammonia, paper containing waste green tea leaves was found to have increased antibacterial activity against Staphylococcus aureus.

Effects of cold immobilization and recovery period on honeybee learning, memory, and responsiveness to sucrose

In addition to human error and variation in laboratory conditions, there are numerous factors that can complicate comparisons among studies. Furthermore, differences in how experimental methods are executed can make it difficult to distinguish between effects of focal versus extraneous variables. Insect neural function is commonly evaluated using Pavlovian conditioning techniques; learning and memory in many species can be assessed using the proboscis extension reflex (PER). However, there are significant inconsistencies in methods used to immobilize insects prior to PER tests. We compared responses of honeybees immobilized in a refrigerator, on ice, and in a freezer, and evaluated influence of recovery interval before testing. Ice-chilling weakly decreased learning (response to an originally neutral odor) more so than refrigeration or freezing, but not 24-h recall of odor. We found no significant differences in responsiveness to sucrose relative to cooling method, but responsiveness was significantly lower among honeybees left to recover for only 0.75 h versus 1.5 or 3 h. Finally, we observed increased responsiveness to sucrose and geraniol between June and August. Our results suggest that inconsistencies in cold immobilization methods could confound interpretation and comparison of results from a large body of work on honeybee learning and memory.

Aversive learning overcomes appetitive innate responding in honeybees

Despite their miniature brain, honeybees have emerged as a powerful model for the study of learning and memory. Yet, they also exhibit innate responses to biologically relevant social signals such as pheromones. Here, we asked whether the bees' developed learning capabilities allow them to overcome hardwired appetitive responses. Can they learn that attractant pheromones, that are not normally associated with a noxious stimulation in nature, predict the punishment of an electric shock? Immobilized honeybees were trained to discriminate two odorants, one that was paired with a shock and another that had no consequences. We measured whether they learned to produce aversive sting extension responses to the punished but not the non-punished odorant. One odorant was a neutral odor without innate value while the other was either an attractive pheromone (geraniol or citral) or an attractive floral odorant (phenylacetaldehyde). In all cases, bees developed a conditioned aversive response to the punished odorant, be it pheromone or not, and efficiently retrieved this information 1h later. No learning asymmetries between odors were found. Thus, associative aversive learning in bees is strong enough to override preprogrammed responding, thus reflecting an impressive behavioral flexibility.