Clint J. Perry

Rapid behavioral maturation accelerates failure of stressed honey bee colonies

Significance Honey bee colony death rates are unsustainably high. While many stressors have been identified that contribute to this problem, we do not know why colonies transition so rapidly from a state of apparent health to failure. It is well known that individual bees react to nutritional and pathogen stresses by foraging precociously: our study explains how colony failure arises from the social responses of individual bees to stress. We used radio tracking to monitor performance of bees and found that workers who begin foraging prematurely perform very poorly. This compounds the stresses on the colony and accelerates failure. We suggest how colonies at risk can be identified early, and the most effective interventions to prevent failure. Many complex factors have been linked to the recent marked increase in honey bee colony failure, including pests and pathogens, agrochemicals, and nutritional stressors. It remains unclear, however, why colonies frequently react to stressors by losing almost their entire adult bee population in a short time, resulting in a colony population collapse. Here we examine the social dynamics underlying such dramatic colony failure. Bees respond to many stressors by foraging earlier in life. We manipulated the demography of experimental colonies to induce precocious foraging in bees and used radio tag tracking to examine the consequences of precocious foraging for their performance. Precocious foragers completed far fewer foraging trips in their life, and had a higher risk of death in their first flights. We constructed a demographic model to explore how this individual reaction of bees to stress might impact colony performance. In the model, when forager death rates were chronically elevated, an increasingly younger forager force caused a positive feedback that dramatically accelerated terminal population decline in the colony. This resulted in a breakdown in division of labor and loss of the adult population, leaving only brood, food, and few adults in the hive. This study explains the social processes that drive rapid depopulation of a colony, and we explore possible strategies to prevent colony failure. Understanding the process of colony failure helps identify the most effective strategies to improve colony resilience.

Associative Mechanisms Allow for Social Learning and Cultural Transmission of String Pulling in an Insect

Social insects make elaborate use of simple mechanisms to achieve seemingly complex behavior and may thus provide a unique resource to discover the basic cognitive elements required for culture, i.e., group-specific behaviors that spread from “innovators” to others in the group via social learning. We first explored whether bumblebees can learn a nonnatural object manipulation task by using string pulling to access a reward that was presented out of reach. Only a small minority “innovated” and solved the task spontaneously, but most bees were able to learn to pull a string when trained in a stepwise manner. In addition, naïve bees learnt the task by observing a trained demonstrator from a distance. Learning the behavior relied on a combination of simple associative mechanisms and trial-and-error learning and did not require “insight”: naïve bees failed a “coiled-string experiment,” in which they did not receive instant visual feedback of the target moving closer when tugging on the string. In cultural diffusion experiments, the skill spread rapidly from a single knowledgeable individual to the majority of a colony’s foragers. We observed that there were several sequential sets (“generations”) of learners, so that previously naïve observers could first acquire the technique by interacting with skilled individuals and, subsequently, themselves become demonstrators for the next “generation” of learners, so that the longevity of the skill in the population could outlast the lives of informed foragers. This suggests that, so long as animals have a basic toolkit of associative and motor learning processes, the key ingredients for the cultural spread of unusual skills are already in place and do not require sophisticated cognition.

Invertebrate learning and cognition: relating phenomena to neural substrate

Diverse invertebrate species have been used for studies of learning and comparative cognition. Although we have gained invaluable information from this, in this study we argue that our approach to comparative learning research is rather deficient. Generally invertebrate learning research has focused mainly on arthropods, and most of that within the Hymenoptera and Diptera. Any true comparative analysis of the distribution of comparative cognitive abilities across phyla is hampered by this bias, and more fundamentally by a reporting bias toward positive results. To understand the limits of learning and cognition for a species, knowing what animals cannot do is at least as important as reporting what they can. Finally, much more effort needs to be focused on the neurobiological analysis of different types of learning to truly understand the differences and similarities of learning types. In this review, we first give a brief overview of the various forms of learning in invertebrates. We also suggest areas where further study is needed for a more comparative understanding of learning. Finally, using what is known of learning in honeybees and the well-studied honeybee brain, we present a model of how various complex forms of learning may be accounted for with the same neural circuitry required for so-called simple learning types. At the neurobiological level, different learning phenomena are unlikely to be independent, and without considering this it is very difficult to correctly interpret the phylogenetic distribution of learning and cognitive abilities. WIREs Cogn Sci 2013, 4:561-582. doi: 10.1002/wcs.1248 For further resources related to this article, please visit the WIREs website.

Unexpected rewards induce dopamine-dependent positive emotion–like state changes in bumblebees

Whether invertebrates exhibit positive emotion–like states and what mechanisms underlie such states remain poorly understood. We demonstrate that bumblebees exhibit dopamine-dependent positive emotion–like states across behavioral contexts. After training with one rewarding and one unrewarding cue, bees that received pretest sucrose responded in a positive manner toward ambiguous cues. In a second experiment, pretest consumption of sucrose solution resulted in a shorter time to reinitiate foraging after a simulated predator attack. These behavioral changes were abolished with topical application of the dopamine antagonist fluphenazine. Further experiments established that pretest sucrose does not simply cause bees to become more exploratory. Our findings present a new opportunity for understanding the fundamental neural elements of emotions and may alter the view of how emotion states affect decision-making in animals.

Honey bees selectively avoid difficult choices

Significance Here we show that honey bees (Apis mellifera) can adaptively alter their behavior in a choice test in response to trial difficulty. Bees preferentially opt out of difficult trials and by doing so, improve their success rate. We discuss whether this choice involves assessing degree of uncertainty (considered a definition of basic metacognition) or whether this task might be solved by associative mechanisms. We propose a hypothesis for how uncertainty might be processed within the known circuitry of the insect brain to frame the concept of uncertainty as a topic for neurobiological analysis. Human decision-making strategies are strongly influenced by an awareness of certainty or uncertainty (a form of metacognition) to increase the chances of making a right choice. Humans seek more information and defer choosing when they realize they have insufficient information to make an accurate decision, but whether animals are aware of uncertainty is currently highly contentious. To explore this issue, we examined how honey bees (Apis mellifera) responded to a visual discrimination task that varied in difficulty between trials. Free-flying bees were rewarded for a correct choice, punished for an incorrect choice, or could avoid choosing by exiting the trial (opting out). Bees opted out more often on difficult trials, and opting out improved their proportion of successful trials. Bees could also transfer the concept of opting out to a novel task. Our data show that bees selectively avoid difficult tasks they lack the information to solve. This finding has been considered as evidence that nonhuman animals can assess the certainty of a predicted outcome, and bees’ performance was comparable to that of primates in a similar paradigm. We discuss whether these behavioral results prove bees react to uncertainty or whether associative mechanisms can explain such findings. To better frame metacognition as an issue for neurobiological investigation, we propose a neurobiological hypothesis of uncertainty monitoring based on the known circuitry of the honey bee brain.

Bumblebees show cognitive flexibility by improving on an observed complex behavior

Very clever bees use tools One hallmark of cognitive complexity is the ability to manipulate objects with a specific goal in mind. Such “tool use” at one time was ascribed to humans alone, but then to primates, next to marine mammals, and later to birds. Now we recognize that many species have the capacity to envision how a particular object might be used to achieve an end. Loukola et al. extend this insight to invertebrates. Bumblebees were trained to see that a ball could be used to produce a reward. These bees then spontaneously rolled the ball when given the chance. Science, this issue p. 833 Bumblebees will use a ball as a tool to obtain a reward. We explored bees’ behavioral flexibility in a task that required transporting a small ball to a defined location to gain a reward. Bees were pretrained to know the correct location of the ball. Subsequently, to obtain a reward, bees had to move a displaced ball to the defined location. Bees that observed demonstration of the technique from a live or model demonstrator learned the task more efficiently than did bees observing a “ghost” demonstration (ball moved via magnet) or without demonstration. Instead of copying demonstrators moving balls over long distances, observers solved the task more efficiently, using the ball positioned closest to the target, even if it was of a different color than the one previously observed. Such unprecedented cognitive flexibility hints that entirely novel behaviors could emerge relatively swiftly in species whose lifestyle demands advanced learning abilities, should relevant ecological pressures arise.

PKCαβγ- and PKCδ-dependent endocytosis of NBCe1-A and NBCe1-B in salivary parotid acinar cells

We examined membrane trafficking of NBCe1-A and NBCe1-B variants of the electrogenic Na(+)-HCO(3)(-) cotransporter (NBCe1) encoded by the SLC4A4 gene, using confocal fluorescent microscopy in rat parotid acinar cells (ParC5 and ParC10). We showed that yellow fluorescent protein (YFP)-tagged NBCe1-A and green fluorescent protein (GFP)-tagged NBCe1-B are colocalized with E-cadherin in the basolateral membrane (BLM) but not with the apical membrane marker zona occludens 1 (ZO-1). We inhibited constitutive recycling with monensin and W13 and detected that NBCe1-A and NBCe1-B accumulated in vesicles marked with the early endosomal marker early endosome antigen-1 (EEA1), with a parallel loss from the BLM. We observed that NBCe1-A and NBCe1-B undergo massive carbachol (CCh)-stimulated redistribution from the BLM into early endosomes. We showed that internalization of NBCe1-A and NBCe1-B was prevented by the general PKC inhibitor GF-109203X, the PKCalphabetagamma-specific inhibitor Gö-6976, and the PKCdelta-specific inhibitor rottlerin. We verified the involvement of PKCdelta by blocking CCh-induced internalization of NBCe1-A-cyan fluorescent protein (CFP) in cells transfected with dominant-negative kinase-dead (Lys376Arg) PKCdelta-GFP. Our data suggest that NBCe1-A and NBCe1-B undergo constitutive and CCh-stimulated endocytosis regulated by conventional PKCs (PKCalphabetagamma) and by novel PKCdelta in rat epithelial cells. To help develop a more complete model of the role of NBCe1 in parotid acinar cells we also investigated the initial phase of the secretory response to cholinergic agonist. In an Ussing chamber study we showed that inhibition of basolateral NBCe1 with 5-chloro-2,3-dihydro-3-(hydroxy-2-thienylmethylene)-2-oxo-1H-indole-1-carboxamide (tenidap) significantly decreases an initial phase of luminal anion secretion measured as a transient short-circuit current (I(sc)) across ParC10 cell monolayers. Using trafficking and functional data we propose a model that describes a physiological role of NBC in salivary acinar cell secretion.

Negative impact of manganese on honeybee foraging

Anthropogenic accumulation of metals such as manganese is a well-established health risk factor for vertebrates. By contrast, the long-term impact of these contaminants on invertebrates is mostly unknown. Here, we demonstrate that manganese ingestion alters brain biogenic amine levels in honeybees and fruit flies. Furthermore, we show that manganese exposure negatively affects foraging behaviour in the honeybee, an economically important pollinator. Our findings indicate that in addition to its direct impact on human health, the common industrial contaminant manganese might also have indirect environmental and economical impacts via the modulation of neuronal and behavioural functions in economically important insects.

Electrogenic NBCe1 (SLC4A4), but not electroneutral NBCn1 (SLC4A7), cotransporter undergoes cholinergic-stimulated endocytosis in salivary ParC5 cells

Cholinergic agonists are major stimuli for fluid secretion in parotid acinar cells. Saliva bicarbonate is essential for maintaining oral health. Electrogenic and electroneutral Na(+)-HCO(3)(-) cotransporters (NBCe1 and NBCn1) are abundant in parotid glands. We previously reported that angiotensin regulates NBCe1 by endocytosis in Xenopus oocytes. Here, we studied cholinergic regulation of NBCe1 and NBCn1 membrane trafficking by confocal fluorescent microscopy and surface biotinylation in parotid epithelial cells. NBCe1 and NBCn1 colocalized with E-cadherin monoclonal antibody at the basolateral membrane (BLM) in polarized ParC5 cells. Inhibition of constitutive recycling with the carboxylic ionophore monensin or the calmodulin antagonist W-13 caused NBCe1 to accumulate in early endosomes with a parallel loss from the BLM, suggesting that NBCe1 is constitutively endocytosed. Carbachol and PMA likewise caused redistribution of NBCe1 from BLM to early endosomes. The PKC inhibitor, GF-109203X, blocked this redistribution, indicating a role for PKC. In contrast, BLM NBCn1 was not downregulated in parotid acinar cells treated with constitutive recycling inhibitors, cholinergic stimulators, or PMA. We likewise demonstrate striking differences in regulation of membrane trafficking of NBCe1 vs. NBCn1 in resting and stimulated cells. We speculate that endocytosis of NBCe1, which coincides with the transition to a steady-state phase of stimulated fluid secretion, could be a part of acinar cell adjustment to a continuous secretory response. Stable association of NBCn1 at the membrane may facilitate constitutive uptake of HCO(3)(-) across the BLM, thus supporting HCO(3)(-) luminal secretion and/or maintaining acid-base homeostasis in stimulated cells.

Accelerated behavioural development changes fine-scale search behaviour and spatial memory in honey bees (Apis mellifera L.)

ABSTRACT Normally, worker honey bees (Apis mellifera) begin foraging when more than 2 weeks old as adults, but if individual bees or the colony is stressed, bees often begin foraging precociously. Here, we examined whether bees that accelerated their behavioural development to begin foraging precociously differed from normal-aged foragers in cognitive performance. We used a social manipulation to generate precocious foragers from small experimental colonies and tested their performance in a free-flight visual reversal learning task, and a test of spatial memory. To assess spatial memory, bees were trained to learn the location of a small sucrose feeder within an array of three landmarks. In tests, the feeder and one landmark were removed and the search behaviour of the bees was recorded. Performance of precocious and normal-aged foragers did not differ in a visual reversal learning task, but the two groups showed a clear difference in spatial memory. Flight behaviour suggested normal-aged foragers were better able to infer the position of the removed landmark and feeder relative to the remaining landmarks than precocious foragers. Previous studies have documented the cognitive decline of old foragers, but this is the first suggestion of a cognitive deficit in young foragers. These data imply that worker honey bees continue their cognitive development during the adult stage. These findings may also help to explain why precocious foragers perform quite poorly as foragers and have a higher than normal loss rate. Summary: Honey bees that begin foraging precociously perform poorly in a test of spatial memory compared with normal-aged foragers, implying there is a cost to this accelerated behavioural development in terms of reduced cognitive ability.