Appetitive Learning: Memories Need Calories

SummaryRecent studies of the way animals learn challenge the idea that food learning relies mainly on how food tastes. Work on Drosophila has now shown that flies must ingest food with a metabolic benefit to form a lasting memory for a learned odour

Toxins induce ‘malaise’ behaviour in the honeybee (Apis mellifera)

To avoid poisoning and death when toxins are ingested, the body responds with a suite of physiological detoxification mechanisms accompanied by behaviours that in mammals often include vomiting, nausea, and lethargy. Few studies have characterised whether insects exhibit characteristic ‘malaise-like’ behaviours in response to intoxication. Here, we used the honeybee to investigate how intoxication produced by injection or ingestion with three toxins with different pharmacological modes of action quinine, amygdalin, and lithium chloride affected behaviour. We found that toxin-induced changes in behaviour were best characterised by more time spent grooming. Bees also had difficulty performing the righting reflex and exhibited specific toxin-induced behaviours such as abdomen dragging and curling up. The expression of these behaviours also depended on whether a toxin had been injected or ingested. When toxins were ingested, they were least 10 times less concentrated in the haemolymph than in the ingested food, suggesting that their absorption through the gut is strongly regulated. Our data show that bees exhibit changes in behaviour that are characteristic of ‘malaise’ and suggest that physiological signalling of toxicosis is accomplished by multiple post-ingestive pathways in animals

B-Vitamins Influence the Consumption of Macronutrients in Honey Bees

Insects require dietary sources of B-vitamins, but relatively little is known about whether they regulate B-vitamin intake in the same way they regulate other nutrients. Honey bees meet their B-vitamin requirements mainly from the pollen they collect. Employing the geometric framework for nutrition, we found that honey bees actively regulate their vitamin intake following Bertrand’s rule. We fed bees with a diet of essential amino acids (EAAs) and carbohydrate (C) to identify how the addition of B-vitamins affected the regulation of these macronutrients. In our experiments, honey bees preferred vitamins in concentrations comparable to those found in honey bee food (pollen, beebread, and royal jelly). Honey bees actively regulated niacin around an optimal value. Supplementing honey bee diets with B-vitamins influenced the amount of EAAs and carbohydrate ingested differently depending on the type of the vitamin. The impact of these vitamins was observed over the course of seven days where honey bees’ mortality increased on diets of low and medium folic acid concentrations. This study provides insights into honey bee food intake regulation and the feeding preferences and sets the basis for future studies considering B-vitamins in honey bees diets

Plant–pollinator interactions and threats to pollination : perspectives from the flower to the landscape

Animal pollinators have driven the diversification of plants on the earth for more than 100 million years. The mutualism between plants and their pollinators rests on an exchange: food for pollinators and efficient vectoring of sexual reproduction for plants. This relationship has been shaped by many factors during the course of evolution. Competition between pollinators for access to nectar and pollen, and among flowering plants for the attention of pollinators, has influenced the shape, colour, and scent of flowers and the extent to which plants invest in the production of pollen and nectar. While many pollinators are ‘generalists’ and visit a variety of flowering plants to obtain food, close relationships between specific pollinators and plant species have developed throughout the course of evolutionary history. An important issue today, given the widespread concern about wild and managed pollinators, is determining how human activities impact the varied relationships between plants and their insect pollinators.The Royal Society and the National Research Foundation of South Africa generously funded the workshop under the SA-UK Scientific Seminars Initiative.http://onlinelibrary.wiley.com/journal/10.1111/(ISSN)1365-24352018-01-31hb2017Zoology and Entomolog

Exposure to acetylcholinesterase inhibitors alters the physiology and motor function of honeybees

Cholinergic signalling is fundamental to neuro-muscular function in most organisms. Sub-lethal doses of neurotoxic pesticides that target cholinergic signalling can alter the behaviour of insects in subtle ways; their influence on non-target organisms may not be readily apparent in simple mortality studies. Beneficial arthropods such as honeybees perform sophisticated behavioural sequences during foraging that, if influenced by pesticides, could impair foraging success and reduce colony health. Here, we investigate the behavioural effects on honeybees of exposure to a selection of pesticides that target cholinergic signalling by inhibiting acetylcholinesterase (AChE). To examine how continued exposure to AChE inhibitors affected motor function, we fed adult foraging worker honeybees sub-lethal concentrations of these compounds in sucrose solution for 24 h. Using an assay for locomotion in bees, we scored walking, stopped, grooming, and upside down behaviour continuously for 15 min. At a 10nM concentration, all the AChE inhibitors caused similar effects on behaviour, notably increased grooming activity and changes in the frequency of bouts of behaviour such as head grooming. Coumaphos caused dose-dependent effects on locomotion as well as grooming behaviour, and a 1µM concentration of coumaphos induced symptoms of malaise such as abdomen grooming and defecation. Biochemical assays confirmed that the 4 compounds we assayed (coumaphos, aldicarb, chlorpyrifos, and donepezil) or their metabolites acted as AChE inhibitors in bees. Furthermore, we show that transcript expression levels of two honeybee acetylcholinesterase inhibitors were selectively upregulated in the brain and in gut tissues in response to AChE inhibitor exposure. The results of our study imply that the effects of pesticides that rely on this mode of action have subtle yet profound effects on physiological effects on behaviour that could lead to reduced survival

Cholinergic pesticides cause mushroom body neuronal inactivation in honeybees

Pesticides that target cholinergic neurotransmission are highly effective, but their use has been implicated in insect pollinator population decline. Honeybees are exposed to two widely used classes of cholinergic pesticide: neonicotinoids (nicotinic receptor agonists) and orga-nophosphate miticides (acetylcholinesterase inhibitors). Although sublethal levels of neoni-cotinoids are known to disrupt honeybee learning and behaviour, the neurophysiological basis of these effects has not been shown. Here, using recordings from mushroom body Kenyon cells in acutely isolated honeybee brain, we show that the neonicotinoids imidacloprid and clothianidin, and the organophosphate miticide coumaphos oxon, cause a depolarization-block of neuronal firing and inhibit nicotinic responses. These effects are observed at concentrations that are encountered by foraging honeybees and within the hive, and are additive with combined application. Our findings demonstrate a neuronal mechanism that may account for the cognitive impairments caused by neonicotinoids, and predict that exposure to multiple pesticides that target cholinergic signalling will cause enhanced toxicity to pollinators

Plant secondary metabolites in nectar: impacts on pollinators and ecological functions

1. The ecological function of secondary metabolites in plant defence, against herbivores is well established, but their role in plant-pollinator interactions is less obvious. Nectar is the major reward for pollinators, so the occurrence of defence compounds in the nectar of many species is unexpected. However, increasing evidence supports a variety of potential benefits for both plant and pollinator from these components. 2. Secondary metabolites in nectar can be toxic or repellent to flower visitors, but they can also go undetected or make nectar attractive . For example, caffeine in nectar improves pollinator memory for cues associated with food rewards and enhances pollen transfer. All of these effects depend on the concentration of nectar metabolites so should be evaluated experimentally at a range of ecologically relevant doses. 3. Beneficial effects may include the following: a) increasing specialization in plant-pollinator interactions, b) protecting nectar from robbery or larceny, and c) preservation of nutrients in nectar from microbial degradation and reducing microbial disease levels in flower visitors. 4. This review synthesises evidence from recent literature that supports selection for secondary metabolites in floral nectar as an adaptation that drives the co-evolution between plants and their pollinators. However, their presence in nectar could simply be a consequence of their occurrence elsewhere in the plant for defence (pleiotropy). We draw attention to the need for studies demonstrating benefits to the plant, the importance of levels of exposure and a effects on target species beyond the current emphasis on alkaloids and bees

Mouthparts of the bumblebee (Bombus terrestris) exhibit poor acuity for the detection of pesticides in nectar

Bees are important pollinators of agricultural crops, but their populations are at risk when pesticides are used. One of the largest risks bees face is poisoning of floral nectar and pollen by insecticides. Studies of bee detection of neonicotinoids have reported contradictory evidence about whether bees can taste these pesticides in sucrose solutions and hence avoid them. Here, we use an assay for the detection of food aversion combined with single-sensillum electrophysiology to test whether the mouthparts of the buff-tailed bumblebee (Bombus terrestris) detect the presence of pesticides in a solution that mimicked the nectar of oilseed rape (Brassica napus). Bees did not avoid consuming solutions containing concentrations of imidacloprid, thiamethoxam, clothianidin, or sulfoxaflor spanning six orders of magnitude, even when these solutions contained lethal doses. Only extremely high concentrations of the pesticides altered spiking in gustatory neurons through a slight reduction in firing rate or change in the rate of adaptation. These data provide strong evidence that bumblebees cannot detect or avoid field-relevant concentrations of pesticides using information from their mouthparts. As bees rarely contact floral nectar with other body parts, we predict that they are at high risk of unwittingly consuming pesticides in the nectar of pesticide-treated crops

Nutrient balancing of the adult worker bumblebee (Bombus terrestris) depends on the dietary source of essential amino acids

Animals carefully regulate the amount of protein that they consume. The quantity of individual essential amino acids (EAAs) obtained from dietary protein depends on the protein source, but how the proportion of EAAs in the diet affects nutrient balancing has rarely been studied. Recent research using the Geometric Framework for Nutrition has revealed that forager honeybees who receive much of their dietary EAAs from floral nectar and not from solid protein have relatively low requirements for dietary EAAs. Here, we examined the nutritional requirements for protein and carbohydrates of foragers of the bufftailed bumblebee Bombus terrestris. By using protein (sodium caseinate) or an equimolar mixture of the 10 EAAs, we found that the intake target (nutritional optimum) of adult workers depended on the source and proportion of dietary EAAs. When bees consumed caseinate-containing diets in a range of ratios between 1:250 and 1:25 (protein to carbohydrate), they achieved an intake target (IT) of 1:149 (w/w). In contrast to those fed protein, bees fed the EAA diets had an IT more biased towards carbohydrates (1:560 w/w) but also had a greater risk of death than those fed caseinate. We also tested how the dietary source of EAAs affected free AAs in bee haemolymph. Bees fed diets near their IT had similar haemolymph AA profiles, whereas bees fed diets high in caseinate had elevated levels of leucine, threonine, valine and alanine in the haemolymph. We found that like honeybees, bumblebee workers prioritize carbohydrate intake and have a relatively low requirement for protein. The dietary source of EAAs influenced both the ratio of protein/EAA to carbohydrate and the overall amount of carbohydrate eaten. Our data support the idea that EAAs and carbohydrates in haemolymph are important determinants of nutritional state in insects.Biotechnology and Biological Sciences Research Council, Natural Environment Research Council, the Wellcome Trust, Department for the Environment, Food, and Rural Affairs, and the Scottish Government under the Insect Pollinators Initiative [BB/I000968/1].http://jeb.biologists.orghb201

The transition from diet to blood: Exploring homeostasis in the insect haemolymph nutrient pool

Nutrition is vital to health, but while the link between diet and body nutritional composition is well explored in humans and other vertebrates, this information is not well understood in insects, despite the vital roles they play in ecosystems, and their increasing use as experimental models. Here we used Nutritional Geometry to explore the rapid physiological response to ingested nutrients in the haemolymph nutritional profile of Spodoptera littoralis caterpillars. We ask whether blood nutrients are maintained homeostatically in the face of variable nutritional intake, or if regulation is more flexible for some nutrients than others (allostasis), which allows animals to adapt to stress by responding in a way that prioritises efficiency of responses in the face of trade‐offs. Caterpillars were placed on 1 of 20 diets, systematically varying in their nutrient ratios (protein: carbohydrate) and density (calorie content), and their consumption was measured. After 48 h, caterpillars were bled, and the macronutrient (protein, carbohydrates and lipids) and nutrient metabolite (amino acids and simple sugars) content of the haemolymph was measured. Proteins comprised 93% of the haemolymph macronutrient pool on average and their concentration increased with protein eaten. The amino acid (AA) pool was dominated by five AAs, and the total pool increased with total nutrient intake. However, the ratio of essential to non‐essential AAs increased as the proportion of protein consumed increased. Carbohydrates were tightly controlled, increasing only on the most extreme carbohydrate intakes. Simple sugars were dominated by glucose and trehalose, and overall, the simple sugar pool showed high levels of homeostasis. Rather than strict homeostasis of blood nutritional properties, an allostatic model seemed to be a better fit for blood nutrient regulation in this generalist herbivore. This flexibility in response to the nutritional composition of the diet may, in part, explain how this species has evolved to extreme dietary generalism and may play a role in its worldwide pest status. Given the range of fitness‐related processes affected by the haemolymph, future studies should examine the physiological impacts of blood nutrient variation on reproduction, growth and response to infection and the trade‐offs between them