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Apis mellifera (honeybees) are a well-established model for the study of learning and cognition. A robust conditioning protocol, the olfactory conditioning of the proboscis extension response (PER), provides a powerful but straightforward method to examine the impact of varying stimuli on learning performance. Herein, we provide a protocol that leverages PER for classroom-based community or student engagement. Specifically, we detail how a class of high school students, as part of the Ryukyu Girls Outreach Program, examined the effects of caffeine and dopamine on learning performance in honeybees. Using a modified version of the PER conditioning protocol, they demonstrated that caffeine, but not dopamine, significantly reduced the number of trials required for a successful conditioning response. In addition to providing an engaging and educational scientific activity, it could be employed, with careful oversight, to garner considerable reliable data examining the effects of varying stimuli on honeybee learning., source:https://www.mdpi.com/2075-4450/12/9/842

BackgroundSublethal exposure to neonicotinoids, a popular class of agricultural pesticides, can lead to behavioral effects that impact the health of pollinators. Therefore, new compounds, such as flupyradifurone (FPF), have recently been developed as 'safer' alternatives. FPF is an excitotoxic nicotinic acetylcholine receptor agonist, similar to neonicotinoids. Given the novelty of FPF, what data exist are focused mostly on assessing the effect of FPF on pollinator mortality. One important avenue for investigation is the potential effect of FPF on the sensitivity of nectar foragers, such as Apis mellifera, to sucrose concentrations. Neonicotinoids can alter this sucrose responsiveness and disrupt foraging. Compounding this effect, neonicotinoid-containing solutions are preferred by A. mellifera over pure sucrose solutions. We therefore conducted four studies, administering FPF under both acute and chronic conditions, and at field-realistic and higher than field-realistic doses, to assess the influence of FPF exposure on sucrose responsiveness and sucrose solutions with FPF in A. mellifera nectar foragers.ResultsWe found no evidence that FPF exposure under acute or chronic field-realistic conditions significantly altered sucrose responsiveness, and we did not find that bees exposed to FPF consumed more of the solution. However, at the much higher median lethal dose (48 h), among bees that survived, FPF-exposed foragers responded to significantly lower concentrations of sucrose than controls and responded at significantly higher rates to all concentrations of sucrose than controls.ConclusionWe found no evidence that FPF alters the sucrose responsiveness of nectar foragers at field-realistic doses during winter or early spring, but caution and further investigation are warranted, particularly on the effects of FPF in conjunction with other stressors. © 2019 Society of Chemical Industry.

BackgroundSublethal exposure to neonicotinoids, a popular class of agricultural pesticides, can lead to behavioral effects that impact the health of pollinators. Therefore, new compounds, such as flupyradifurone (FPF), have recently been developed as 'safer' alternatives. FPF is an excitotoxic nicotinic acetylcholine receptor agonist, similar to neonicotinoids. Given the novelty of FPF, what data exist are focused mostly on assessing the effect of FPF on pollinator mortality. One important avenue for investigation is the potential effect of FPF on the sensitivity of nectar foragers, such as Apis mellifera, to sucrose concentrations. Neonicotinoids can alter this sucrose responsiveness and disrupt foraging. Compounding this effect, neonicotinoid-containing solutions are preferred by A. mellifera over pure sucrose solutions. We therefore conducted four studies, administering FPF under both acute and chronic conditions, and at field-realistic and higher than field-realistic doses, to assess the influence of FPF exposure on sucrose responsiveness and sucrose solutions with FPF in A. mellifera nectar foragers.ResultsWe found no evidence that FPF exposure under acute or chronic field-realistic conditions significantly altered sucrose responsiveness, and we did not find that bees exposed to FPF consumed more of the solution. However, at the much higher median lethal dose (48 h), among bees that survived, FPF-exposed foragers responded to significantly lower concentrations of sucrose than controls and responded at significantly higher rates to all concentrations of sucrose than controls.ConclusionWe found no evidence that FPF alters the sucrose responsiveness of nectar foragers at field-realistic doses during winter or early spring, but caution and further investigation are warranted, particularly on the effects of FPF in conjunction with other stressors. © 2019 Society of Chemical Industry.

In several insects, sex-pheromones are essential for reproduction and reproductive isolation. Pheromones generally elicit stereotyped behaviors. In moths, these are attraction to conspecific sex-pheromone sources and deterrence for heterospecific sex-pheromone. Contrasting with these innate behaviors, some results in social insects point toward effects of non-sex-pheromones on perception and learning. We report the effects of sex-pheromone pre-exposure on gustatory perception and habituation (a non-associative learning) in male Agrotis ipsilon moths, a non-social insect. We also studied the effect of Z5-decenyl acetate (Z5), a compound of the sex-pheromone of the related species Agrotis segetum. We hypothesized that conspecific sex-pheromone and Z5 would have opposite effects. Pre-exposure to either the conspecific sex-pheromone or Z5 lasted 15 min and was done either immediately or 24 h before the experiments, using their solvent alone (hexane) as control. In a sucrose responsiveness assay, pre-exposure to the conspecific sex-pheromone had no effect on the dose-response curve at either delays. By contrast, Z5 slightly improved sucrose responsiveness 15 min but not 24 h after pre-exposure. Interestingly, the conspecific sex-pheromone and Z5 had time-dependent effects on gustatory habituation: pre-exposing the moths with Z5 hindered learning after immediate but not 24-h pre-exposure, whereas pre-exposure to the conspecific sex-pheromone hindered learning at 24-h but not immediate pre-exposure. They did not have opposite effects. This is the first time a sex-pheromone is reported to affect learning in a non-social insect. The difference in modulation between conspecific sex-pheromone and Z5 suggests that con- and hetero-specific sex-pheromones act on plasticity through different cerebral pathways.

In several insects, sex-pheromones are essential for reproduction and reproductive isolation. Pheromones generally elicit stereotyped behaviors. In moths, these are attraction to conspecific sex-pheromone sources and deterrence for heterospecific sex-pheromone. Contrasting with these innate behaviors, some results in social insects point toward effects of non-sex-pheromones on perception and learning. We report the effects of sex-pheromone pre-exposure on gustatory perception and habituation (a non-associative learning) in male Agrotis ipsilon moths, a non-social insect. We also studied the effect of Z5-decenyl acetate (Z5), a compound of the sex-pheromone of the related species Agrotis segetum. We hypothesized that conspecific sex-pheromone and Z5 would have opposite effects. Pre-exposure to either the conspecific sex-pheromone or Z5 lasted 15 min and was done either immediately or 24 h before the experiments, using their solvent alone (hexane) as control. In a sucrose responsiveness assay, pre-exposure to the conspecific sex-pheromone had no effect on the dose-response curve at either delays. By contrast, Z5 slightly improved sucrose responsiveness 15 min but not 24 h after pre-exposure. Interestingly, the conspecific sex-pheromone and Z5 had time-dependent effects on gustatory habituation: pre-exposing the moths with Z5 hindered learning after immediate but not 24-h pre-exposure, whereas pre-exposure to the conspecific sex-pheromone hindered learning at 24-h but not immediate pre-exposure. They did not have opposite effects. This is the first time a sex-pheromone is reported to affect learning in a non-social insect. The difference in modulation between conspecific sex-pheromone and Z5 suggests that con- and hetero-specific sex-pheromones act on plasticity through different cerebral pathways.

In this brief review, I argue that the impact of a stimulus on behavioral control increase as the distance of the stimulus to the body decreases. Habituation, i.e., decrement in response intensity repetition of the triggering stimulus, is the default state for sensory processing, and the likelihood of habituation is higher for distal stimuli. Sensitization, i.e., increment in response intensity upon stimulus repetition, occurs in a state dependent manner for proximal stimuli that make direct contact with the body. In Pavlovian conditioning paradigms, the unconditioned stimulus (US) is always a more proximal stimulus than the conditioned stimulus (CS). The mechanisms of associative and non-associative learning are not independent. CS?US pairings lead to formation of associations if sensitizing modulation from a proximal US prevents the habituation for a distal anticipatory CS. Identification of the necessary and sufficient conditions for the formation of associations has been a driving influence on learning theory and research. In Pavlovian conditioning, a conditioned stimulus (CS) acquires the ability to trigger a new response by virtue of being paired with an unconditioned stimulus (US), which by definition is biologically important and capable of triggering an innate reflex. Starting with British associationism, early theories of conditioning were based on the premise that temporal contiguity was both necessary and sufficient for stimulus associations (Gormezano and Kehoe, 1981). Although the temporal coincidence of the CS?US pair is still accepted to be necessary, research since late 1960's presented irrevocable challenges to its sufficiency for the formation of associations (Durlach, 1989). Of particular importance was the discovery of blocking (Kamin, 1968), where an association fails to be formed in spite of the seamless temporal contiguity between the CS and the US, if the CS is presented in a compound with another CS that had previously been associated with

In this brief review, I argue that the impact of a stimulus on behavioral control increase as the distance of the stimulus to the body decreases. Habituation, i.e., decrement in response intensity repetition of the triggering stimulus, is the default state for sensory processing, and the likelihood of habituation is higher for distal stimuli. Sensitization, i.e., increment in response intensity upon stimulus repetition, occurs in a state dependent manner for proximal stimuli that make direct contact with the body. In Pavlovian conditioning paradigms, the unconditioned stimulus (US) is always a more proximal stimulus than the conditioned stimulus (CS). The mechanisms of associative and non-associative learning are not independent. CS?US pairings lead to formation of associations if sensitizing modulation from a proximal US prevents the habituation for a distal anticipatory CS. Identification of the necessary and sufficient conditions for the formation of associations has been a driving influence on learning theory and research. In Pavlovian conditioning, a conditioned stimulus (CS) acquires the ability to trigger a new response by virtue of being paired with an unconditioned stimulus (US), which by definition is biologically important and capable of triggering an innate reflex. Starting with British associationism, early theories of conditioning were based on the premise that temporal contiguity was both necessary and sufficient for stimulus associations (Gormezano and Kehoe, 1981). Although the temporal coincidence of the CS?US pair is still accepted to be necessary, research since late 1960's presented irrevocable challenges to its sufficiency for the formation of associations (Durlach, 1989). Of particular importance was the discovery of blocking (Kamin, 1968), where an association fails to be formed in spite of the seamless temporal contiguity between the CS and the US, if the CS is presented in a compound with another CS that had previously been associated with

In several insects, sex-pheromones are essential for reproduction and reproductive isolation. Pheromones generally elicit stereotyped behaviors. In moths, these are attraction to conspecific sex-pheromone sources and deterrence for heterospecific sex-pheromone. Contrasting with these innate behaviors, some results in social insects point toward effects of non-sex-pheromones on perception and learning. We report the effects of sex-pheromone pre-exposure on gustatory perception and habituation (a non-associative learning) in male Agrotis ipsilon moths, a non-social insect. We also studied the effect of Z5-decenyl acetate (Z5), a compound of the sex-pheromone of the related species Agrotis segetum. We hypothesized that conspecific sex-pheromone and Z5 would have opposite effects. Pre-exposure to either the conspecific sex-pheromone or Z5 lasted 15 min and was done either immediately or 24 h before the experiments, using their solvent alone (hexane) as control. In a sucrose responsiveness assay, pre-exposure to the conspecific sex-pheromone had no effect on the dose-response curve at either delays. By contrast, Z5 slightly improved sucrose responsiveness 15 min but not 24 h after pre-exposure. Interestingly, the conspecific sex-pheromone and Z5 had time-dependent effects on gustatory habituation: pre-exposing the moths with Z5 hindered learning after immediate but not 24-h pre-exposure, whereas pre-exposure to the conspecific sex-pheromone hindered learning at 24-h but not immediate pre-exposure. They did not have opposite effects. This is the first time a sex-pheromone is reported to affect learning in a non-social insect. The difference in modulation between conspecific sex-pheromone and Z5 suggests that con- and hetero-specific sex-pheromones act on plasticity through different cerebral pathways.

Adult honey bees are maintained in vitro in laboratory cages for a variety of purposes. For example, researchers may wish to perform experiments on honey bees caged individually or in groups to study aspects of parasitology, toxicology, or physiology under highly controlled conditions, or they may cage whole frames to obtain freshly emerged workers of known age cohorts. Regardless of purpose, researchers must manage a number of variables, ranging from selection of study subjects (e.g. honey bee subspecies) to experimental environment (e.g. temperature and relative humidity). Although decisions made by researchers may not necessarily jeopardize the scientific rigour of an experiment, they may profoundly affect results, and may make comparisons with similar, but independent, studies difficult. Focusing primarily on workers, we provide recommendations for maintaining adults under in vitro laboratory conditions, whilst acknowledging gaps in our understanding that require further attention. We specifically describe how to properly obtain honey bees, and how to choose appropriate cages, incubator conditions, and food to obtain biologically relevant and comparable experimental results. Additionally, we provide broad recommendations for experimental design and statistical analyses of data that arises from experiments using caged honey bees. The ultimate goal of this, and of all COLOSS BEEBOOK papers, is not to stifle science with restrictions, but rather to provide researchers with the appropriate tools to generate comparable data that will build upon our current understanding of honey bees

Young (7-day-old), middle-aged (28-day-old), and old (49-day-old) Drosophila melanogaster were compared for acquisition, then extinction, of excitatory conditioning of the Proboscis Extension Response. The contribution of nonassociative processes (sucrose-induced Central Excitatory State, Pseudoconditioning) in the elicitation of conditioned responses was simultaneously assessed. Old flies displayed a faster and stronger acquisition than middle-aged ones, whereas no significant acquisition was stated in young flies. The influence of nonassociative processes on the acquisition was of minor importance, even though Central Excitatory State increased with age. Old flies proved also to be the slowest to extinguish. Because nonassociative contamination was not involved during extinction, one may wonder whether extinction was delayed in old flies mainly due to a higher level of acquisition or to a behavioral rigidity. (C) 1999 Elsevier Science Inc.