Evidence for socially influenced and potentially actively coordinated cooperation by bumblebees.

Cooperation is common in animals, yet the specific mechanisms driving collaborative behaviour in different species remain unclear. We investigated the proximate mechanisms underlying the cooperative behaviour of bumblebees in two different tasks, where bees had to simultaneously push a block in an arena or a door at the end of a tunnel for access to reward. In both tasks, when their partner's entry into the arena/tunnel was delayed, bees took longer to first push the block/door compared with control bees that learned to push alone. In the tunnel task, just before gaining access to reward, bees were more likely to face towards their partner than expected by chance or compared with controls. These results show that bumblebees' cooperative behaviour is not simply a by-product of individual efforts but is socially influenced. We discuss how bees' turning behaviours, e.g. turning around before first reaching the door when their partner was delayed and turning back towards the door in response to seeing their partner heading towards the door, suggest the potential for active coordination. However, because these behaviours could also be interpreted as combined responses to social and secondary reinforcement cues, future studies are needed to help clarify whether bumblebees truly use active coordination.

The role of cognition in nesting.

For many animals, nests are essential for reproductive success. Nesting individuals need to carry out a range of potentially challenging tasks, from selecting an appropriate site and choosing suitable materials to constructing the nest and defending it against competitors, parasites and predators. Given the high fitness stakes involved, and the diverse impacts both the abiotic and social environment can have on nesting success, we might expect cognition to facilitate nesting efforts. This should be especially true under variable environmental conditions, including those changing due to anthropogenic impacts. Here, we review, across a wide range of taxa, evidence linking cognition to nesting behaviours, including selection of nesting sites and materials, nest construction, and nest defence. We also discuss how different cognitive abilities may increase an individual's nesting success. Finally, we highlight how combining experimental and comparative research can uncover the links between cognitive abilities, nesting behaviours and the evolutionary pathways that may have led to the associations between them. In so doing, the review highlights current knowledge gaps and provides suggestions for future research. This article is part of the theme issue 'The evolutionary ecology of nests: a cross-taxon approach'.

Glyphosate and a glyphosate-based herbicide affect bumblebee gut microbiota.

Pollinator decline is one of the gravest challenges facing the world today, and the overuse of pesticides may be among its causes. Here, we studied whether glyphosate, the world's most widely used pesticide, affects the bumblebee gut microbiota. We exposed the bumblebee diet to glyphosate and a glyphosate-based herbicide and quantified the microbiota community shifts using 16S rRNA gene sequencing. Furthermore, we estimated the potential sensitivity of bee gut microbes to glyphosate based on previously reported presence of target enzyme. Glyphosate increased, whereas the glyphosate-based herbicide decreased gut microbiota diversity, indicating that negative effects are attributable to co-formulants. Both glyphosate and the glyphosate-based herbicide treatments significantly decreased the relative abundance of potentially glyphosate-sensitive bacterial species Snodgrasella alvi. However, the relative abundance of potentially glyphosate-sensitive Candidatus Schmidhempelia genera increased in bumblebees treated with glyphosate. Overall, 50% of the bacterial genera detected in the bee gut microbiota were classified as potentially resistant to glyphosate, while 36% were classified as sensitive. Healthy core microbiota have been shown to protect bees from parasite infections, change metabolism, and decrease mortality. Thus, the heavy use of glyphosate-based herbicides may have implications on bees and ecosystems.

Oral exposure to thiacloprid-based pesticide (Calypso SC480) causes physical poisoning symptoms and impairs the cognitive abilities of bumble bees.

BACKGROUND: Pesticides are identified as one of the major reasons for the global pollinator decline. However, the sublethal effects of pesticide residue levels found in pollen and nectar on pollinators have been studied little. The aim of our research was to study whether oral exposure to the thiacloprid levels found in pollen and nectar affect the learning and long-term memory of bumble bees. We tested the effects of two exposure levels of thiacloprid-based pesticide (Calypso SC480) on buff-tailed bumble bee (Bombus terrestris) in laboratory utilizing a learning performance and memory tasks designed to be difficult enough to reveal large variations across the individuals. RESULTS: The lower exposure level of the thiacloprid-based pesticide impaired the bees' learning performance but not long-term memory compared to the untreated controls. The higher exposure level caused severe acute symptoms, due to which we were not able to test the learning and memory. CONCLUSIONS: Our results show that oral exposure to a thiacloprid-based pesticide, calculated based on residue levels found in pollen and nectar, not only causes sublethal effects but also acute lethal effects on bumble bees. Our study underlines an urgent demand for better understanding of pesticide residues in the environment, and of the effects of those residue levels on pollinators. These findings fill the gap in the existing knowledge and help the scientific community and policymakers to enhance the sustainable use of pesticides.

Ecological and evolutionary consequences of selective interspecific information use.

Recent work has shown that animals frequently use social information from individuals of their own species as well as from other species; however, the ecological and evolutionary consequences of this social information use remain poorly understood. Additionally, information users may be selective in their social information use, deciding from whom and how to use information, but this has been overlooked in an interspecific context. In particular, the intentional decision to reject a behaviour observed via social information has received less attention, although recent work has indicated its presence in various taxa. Based on existing literature, we explore in which circumstances selective interspecific information use may lead to different ecological and coevolutionary outcomes between two species, such as explaining observed co-occurrences of putative competitors. The initial ecological differences and the balance between the costs of competition and the benefits of social information use potentially determine whether selection may lead to trait divergence, convergence or coevolutionary arms race between two species. We propose that selective social information use, including adoption and rejection of behaviours, may have far-reaching fitness consequences, potentially leading to community-level eco-evolutionary outcomes. We argue that these consequences of selective interspecific information use may be much more widespread than has thus far been considered.

Field-realistic acute exposure to glyphosate-based herbicide impairs fine-color discrimination in bumblebees.

Pollinator decline is a grave challenge worldwide. One of the main culprits for this decline is the widespread use of, and pollinators' chronic exposure to, agrochemicals. Here, we examined the effect of a field-realistic dose of the world's most commonly used pesticide, glyphosate-based herbicide (GBH), on bumblebee cognition. We experimentally tested bumblebee (Bombus terrestris) color and scent discrimination using acute GBH exposure, approximating a field-realistic dose from a day's foraging in a patch recently sprayed with GBH. In a 10-color discrimination experiment with five learning bouts, GBH treated bumblebees' learning rate fell to zero by third learning bout, whereas the control bees increased their performance in the last two bouts. In the memory test, the GBH treated bumblebees performed to near chance level, indicating that they had lost everything they had learned during the learning bouts, while the control bees were performing close to the level in their last learning bout. However, GBH did not affect bees' learning in a 2-color or 10-odor discrimination experiment, which suggests that the impact is limited to fine color learning and does not necessarily generalize to less specific tasks or other modalities. These results indicate that the widely used pesticide damages bumblebees' fine-color discrimination, which is essential to the pollinator's individual success and to colony fitness in complex foraging environments. Hence, our study suggests that acute sublethal exposure to GBH poses a greater threat to pollination-based ecosystem services than previously thought, and that tests for learning and memory should be integrated into pesticide risk assessment.

Prior associations affect bumblebees' generalization performance in a tool-selection task.

A small brain and short life allegedly limit cognitive abilities. Our view of invertebrate cognition may also be biased by the choice of experimental stimuli. Here, the stimuli (color) pairs used in the match-to-sample tasks affected the performance of buff-tailed bumblebees (Bombus terrestris). We trained the bees to roll a tool, a ball, to a goal that matched its color. Bees trained with a yellow-and-orange/red stimuli pair took more training bouts to reach our color-matching criterion than those trained with a blue-and-yellow stimuli pair. When assessing the bees' concept learning ability in a transfer test with a novel color, the bees trained with blue and yellow (novel color: orange/red) were highly successful, the bees trained with blue and orange/red (novel color: yellow) did not differ from random, and those trained with yellow and orange/red (novel color: blue) failed the test. These results highlight that stimulus salience can affect our conclusions on test subjects' cognitive ability. Therefore, we encourage paying attention to stimulus salience (among other factors) when assessing the cognition of invertebrates.

Illusional Perspective across Humans and Bees.

For two centuries, visual illusions have attracted the attention of neurobiologists and comparative psychologists, given the possibility of investigating the complexity of perceptual mechanisms by using relatively simple patterns. Animal models, such as primates, birds, and fish, have played a crucial role in understanding the physiological circuits involved in the susceptibility of visual illusions. However, the comprehension of such mechanisms is still a matter of debate. Despite their different neural architectures, recent studies have shown that some arthropods, primarily Hymenoptera and Diptera, experience illusions similar to those humans do, suggesting that perceptual mechanisms are evolutionarily conserved among species. Here, we review the current state of illusory perception in bees. First, we introduce bees' visual system and speculate which areas might make them susceptible to illusory scenes. Second, we review the current state of knowledge on misperception in bees (Apidae), focusing on the visual stimuli used in the literature. Finally, we discuss important aspects to be considered before claiming that a species shows higher cognitive ability while equally supporting alternative hypotheses. This growing evidence provides insights into the evolutionary origin of visual mechanisms across species.

Conceptual preferences can be transmitted via selective social information use between competing wild bird species.

Concept learning is considered a high-level adaptive ability. Thus far, it has been studied in laboratory via asocial trial and error learning. Yet, social information use is common among animals but it remains unknown whether concept learning by observing others occurs. We tested whether pied flycatchers (Ficedula hypoleuca) form conceptual relationships from the apparent choices of nest-site characteristics (geometric symbol attached to the nest-box) of great tits (Parus major). Each wild flycatcher female (n = 124) observed one tit pair that exhibited an apparent preference for either a large or a small symbol and was then allowed to choose between two nest-boxes with a large and a small symbol, but the symbol shape was different to that on the tit nest. Older flycatcher females were more likely to copy the symbol size preference of tits than yearling flycatcher females when there was a high number of visible eggs or a few partially visible eggs in the tit nest. However, this depended on the phenotype, copying switched to rejection as a function of increasing body size. Possibly the quality of and overlap in resource use with the tits affected flycatchers' decisions. Hence, our results suggest that conceptual preferences can be horizontally transmitted across coexisting animals, which may increase the performance of individuals that use concept learning abilities in their decision-making.

Pesticide residues in honeybee-collected pollen: does the EU regulation protect honeybees from pesticides?

Researchers globally identify pesticides as one of the main reasons for pollinator decline. In the European Union (EU), extensive legislation is implemented to protect pollinators from harmful pesticide exposure. The aim of our study was to discover whether the pesticide residue levels in honeybee matrices, such as nectar and pollen, exceeded the chronic or acute toxicity levels when beehives were located next to fields treated with specific insecticides. The insecticides were used according to the EU legislation and its national implementation. The experiments were conducted in turnip rape, oilseed rape, and caraway fields in southern Finland during the years 2019 and 2020. The pesticides used in the experiments contained the active substances lambda-cyhalothrin (2019), esfenvalerate (2020), and tau-fluvalinate (2020). However, the honeybee-collected pollen and nectar were analyzed for residues of more than 100 active substances. The results showed that the pesticide residue levels clearly remained under the oral acute toxicity for honeybees, although we found high levels of thiacloprid residues in the pollen collected in 2019. The pesticide residues in nectar were below LOQ values, which was most likely due to the rainy weather conditions together with the chosen sampling method. No statistically significant differences were observed between the insecticide-treated and untreated fields. In light of our research, the EU legislation protected honeybees from oral acute toxicity during the years 2019 and 2020. However, potential sublethal effects of thiacloprid and other pesticide compounds found in the collected pollen cannot be ruled out. In the future, constant monitoring of pesticide exposure of honeybees and wild pollinators should be established to ensure that pesticide legislation, and its implementation across the EU successfully protects pollinators and their services in agricultural environments.

Quantitative abilities of invertebrates: a methodological review.

Quantitative abilities are widely recognized to play important roles in several ecological contexts, such as foraging, mate choice, and social interaction. Indeed, such abilities are widespread among vertebrates, in particular mammals, birds, and fish. Recently, there has been an increasing number of studies on the quantitative abilities of invertebrates. In this review, we present the current knowledge in this field, especially focusing on the ecological relevance of the capacity to process quantitative information, the similarities with vertebrates, and the different methods adopted to investigate this cognitive skill. The literature argues, beyond methodological differences, a substantial similarity between the quantitative abilities of invertebrates and those of vertebrates, supporting the idea that similar ecological pressures may determine the emergence of similar cognitive systems even in distantly related species.

Non-numerical strategies used by bees to solve numerical cognition tasks.

We examined how bees solve a visual discrimination task with stimuli commonly used in numerical cognition studies. Bees performed well on the task, but additional tests showed that they had learned continuous (non-numerical) cues. A network model using biologically plausible visual feature filtering and a simple associative rule was capable of learning the task using only continuous cues inherent in the training stimuli, with no numerical processing. This model was also able to reproduce behaviours that have been considered in other studies indicative of numerical cognition. Our results support the idea that a sense of magnitude may be more primitive and basic than a sense of number. Our findings highlight how problematic inadvertent continuous cues can be for studies of numerical cognition. This remains a deep issue within the field that requires increased vigilance and cleverness from the experimenter. We suggest ways of better assessing numerical cognition in non-speaking animals, including assessing the use of all alternative cues in one test, using cross-modal cues, analysing behavioural responses to detect underlying strategies, and finding the neural substrate.

Foraging Bumblebees Selectively Attend to Other Types of Bees Based on Their Reward-Predictive Value.

Using social information can be an efficient strategy for learning in a new environment while reducing the risks associated with trial-and-error learning. Whereas social information from conspecifics has long been assumed to be preferentially attended by animals, heterospecifics can also provide relevant information. Because different species may vary in their informative value, using heterospecific social information indiscriminately can be ineffective and even detrimental. Here, we evaluated how selective use of social information might arise at a proximate level in bumblebees (Bombus terrestris) as a result of experience with demonstrators differing in their visual appearance and in their informative value as reward predictors. Bumblebees were first trained to discriminate rewarding from unrewarding flowers based on which type of "heterospecific" (one of two differently painted model bees) was next to each flower. Subsequently, these bumblebees were exposed to a novel foraging context with two live painted bees. In this novel context, observer bumblebees showed significantly more social information-seeking behavior towards the type of bees that had predicted reward during training. Bumblebees were not attracted by paint-marked small wooden balls (moved via magnets) or paint-marked non-pollinating heterospecifics (woodlice; Porcellio laevis) in the novel context, indicating that bees did not simply respond to conditioned color cues nor to irrelevant social cues, but rather had a "search image" of what previously constituted a valuable, versus invaluable, information provider. The behavior of our bumblebees suggests that their use of social information is governed by learning, is selective, and extends beyond conspecifics.

Bumblebees Use Sequential Scanning of Countable Items in Visual Patterns to Solve Numerosity Tasks.

Most research in comparative cognition focuses on measuring if animals manage certain tasks; fewer studies explore how animals might solve them. We investigated bumblebees' scanning strategies in a numerosity task, distinguishing patterns with two items from four and one from three, and subsequently transferring numerical information to novel numbers, shapes, and colors. Video analyses of flight paths indicate that bees do not determine the number of items by using a rapid assessment of number (as mammals do in "subitizing"); instead, they rely on sequential enumeration even when items are presented simultaneously and in small quantities. This process, equivalent to the motor tagging ("pointing") found for large number tasks in some primates, results in longer scanning times for patterns containing larger numbers of items. Bees used a highly accurate working memory, remembering which items have already been scanned, resulting in fewer than 1% of re-inspections of items before making a decision. Our results indicate that the small brain of bees, with less parallel processing capacity than mammals, might constrain them to use sequential pattern evaluation even for low quantities.

Bumblebees show cognitive flexibility by improving on an observed complex behavior.

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.

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

[This corrects the article DOI: 10.1371/journal.pbio.1002564.].

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.

Interspecific variation in the relationship between clutch size, laying date and intensity of urbanization in four species of hole-nesting birds.

The increase in size of human populations in urban and agricultural areas has resulted in considerable habitat conversion globally. Such anthropogenic areas have specific environmental characteristics, which influence the physiology, life history, and population dynamics of plants and animals. For example, the date of bud burst is advanced in urban compared to nearby natural areas. In some birds, breeding success is determined by synchrony between timing of breeding and peak food abundance. Pertinently, caterpillars are an important food source for the nestlings of many bird species, and their abundance is influenced by environmental factors such as temperature and date of bud burst. Higher temperatures and advanced date of bud burst in urban areas could advance peak caterpillar abundance and thus affect breeding phenology of birds. In order to test whether laying date advance and clutch sizes decrease with the intensity of urbanization, we analyzed the timing of breeding and clutch size in relation to intensity of urbanization as a measure of human impact in 199 nest box plots across Europe, North Africa, and the Middle East (i.e., the Western Palearctic) for four species of hole-nesters: blue tits (Cyanistes caeruleus), great tits (Parus major), collared flycatchers (Ficedula albicollis), and pied flycatchers (Ficedula hypoleuca). Meanwhile, we estimated the intensity of urbanization as the density of buildings surrounding study plots measured on orthophotographs. For the four study species, the intensity of urbanization was not correlated with laying date. Clutch size in blue and great tits does not seem affected by the intensity of urbanization, while in collared and pied flycatchers it decreased with increasing intensity of urbanization. This is the first large-scale study showing a species-specific major correlation between intensity of urbanization and the ecology of breeding. The underlying mechanisms for the relationships between life history and urbanization remain to be determined. We propose that effects of food abundance or quality, temperature, noise, pollution, or disturbance by humans may on their own or in combination affect laying date and/or clutch size.

Species and abundance of ectoparasitic flies (Diptera) in pied flycatcher nests in Fennoscandia.

BACKGROUND: Birds host several ectoparasitic fly species with negative effects on nestling health and reproductive output, and with the capability of transmitting avian blood parasites. Information on the abundance and distribution of the ectoparasitic fly genera Ornithomya (Hippoboscidae) and Protocalliphora (Calliphoridae) in northern Europe is still generally poor, and we thus explored their geographic range and occurrence of these flies in the nests of a common avian model species, the pied flycatcher Ficedula hypoleuca. METHODS: Nests of F. hypoleuca were collected from 21 locations across Fennoscandia in summer 2013, across a latitudinal gradient (between 56 °N - 70 °N) and examined for the presence of fly puparia. Adult specimens of Ornithomya spp. were also collected for species identification. Fly species were identified morphologically and identifications confirmed with DNA barcoding. RESULTS: We found three species: two louse-flies - Ornithomya chloropus and O. avicularia - and one blow-fly, Protocalliphora azurea. The prevalence of O. avicularia was higher in southern latitudes and this species was not encountered beyond 62 °N whereas O. chloropus and P. azurea occurred across the whole range of latitudes. The prevalence of O. chloropus further increased with increasing distance from the coast - a pattern not documented before. The three fly species showed no interspecific associations in their prevalence. CONCLUSIONS: Our study revealed relatively high prevalence for all the species (O. chloropus 59 %, O. avicularia 20 %, P. azurea 32 %), and an interesting spatial pattern in the prevalence of the two louse fly species. Our sample did not indicate any major range shifts towards the north for the southern species as compared to the information from the past. Morphological identification of O. chloropus did not match with the corresponding sequences published in the GenBank and taxonomy of this group calls for further studies.

Variation in clutch size in relation to nest size in birds.

Nests are structures built to support and protect eggs and/or offspring from predators, parasites, and adverse weather conditions. Nests are mainly constructed prior to egg laying, meaning that parent birds must make decisions about nest site choice and nest building behavior before the start of egg-laying. Parent birds should be selected to choose nest sites and to build optimally sized nests, yet our current understanding of clutch size-nest size relationships is limited to small-scale studies performed over short time periods. Here, we quantified the relationship between clutch size and nest size, using an exhaustive database of 116 slope estimates based on 17,472 nests of 21 species of hole and non-hole-nesting birds. There was a significant, positive relationship between clutch size and the base area of the nest box or the nest, and this relationship did not differ significantly between open nesting and hole-nesting species. The slope of the relationship showed significant intraspecific and interspecific heterogeneity among four species of secondary hole-nesting species, but also among all 116 slope estimates. The estimated relationship between clutch size and nest box base area in study sites with more than a single size of nest box was not significantly different from the relationship using studies with only a single size of nest box. The slope of the relationship between clutch size and nest base area in different species of birds was significantly negatively related to minimum base area, and less so to maximum base area in a given study. These findings are consistent with the hypothesis that bird species have a general reaction norm reflecting the relationship between nest size and clutch size. Further, they suggest that scientists may influence the clutch size decisions of hole-nesting birds through the provisioning of nest boxes of varying sizes.