Injury shortens life expectancy in ants and affects some risk-related decisions of workers.

The Formica cinerea ants are known to be highly territorial and aggressively defend their nest and foraging areas against other ants. During the foraging, workers engage in large-scale battles with other colonies of ants and injuries often occur in the process. Such injuries open the body up to pathologies and can lead to costs expressed in lower survival. Here, we addressed the significance of injury in dictating decisions related to engagement in risky behavior in ants (i.e., rescue and aggression). We manipulated the life expectancies of F. cinerea workers by injury and found that the survival of injured workers was shorter compared to the intact individuals. Furthermore, we found that injured workers discriminated between the intact and injured nestmates and showed more rescue behavior toward intact individuals. These rescue actions were expressed as digging around the trapped ant in need of rescue, pulling at its body parts, transporting the sand covering it, and biting the thread entrapping it. In turn, intact and injured workers showed similar and high levels of aggression toward heterospecifics. Our findings highlight the role of behavioral context in the studies devoted to the decision-making processes among social insects and the importance of life expectancy in their behavioral patterns.

Operant conditioning in antlion larvae and its impairment following exposure to elevated temperatures.

Although ambush predators were previously considered limited in their cognitive abilities compared to their widely foraging relatives, there is accumulating evidence it does not hold true. Pit-building antlions are already known to associate vibrations in the sand with the arrival of prey. We used a T-maze and successfully trained antlions to turn right or left against their initial turning bias, leading to a suitable substrate for digging traps. We present here the first evidence for operant conditioning and T-maze solving in antlions. Furthermore, we show that exposure of second instar larvae to an elevated temperature led to impaired retention of what was learned in a T-maze when tested after moulting into the third instar, compared to larvae raised under a more benign temperature. We suggest that climate change, involving an increase in mean temperatures as well as rare events (e.g., heatwaves) might negatively affect the retention of operant conditioning in antlions, alongside known, more frequently studied effects, such as changes in body size and distribution.

Discontinued alcohol consumption elicits withdrawal symptoms in honeybees.

The honeybee continues to be developed as a model species in many research areas, including studies related to the effects of alcohol. Here, we investigate whether workers display one of the key features of alcoholism, namely withdrawal symptoms. We show that workers fed for a prolonged time on food spiked with ethanol, after discontinuation of access to such food, exhibited a marked increase in the consumption of ethanol and a slight increase in mortality. We additionally show that withdrawal symptoms do not include an increase in appetitiveness of ethanol diluted in water. Our results demonstrate that workers can develop alcohol dependence, which might be especially important in the natural setting of repeated exposure to ethanol in floral nectar and for their potential as a model of alcohol addiction.

Larval antlions show a cognitive ability/hunting efficiency trade-off connected with the level of behavioural asymmetry.

Recently, antlion larvae with greater behavioural asymmetry were shown to have improved learning abilities. However, a major evolutionary question that remained unanswered was why this asymmetry does not increase in all individuals during development. Here, we show that a trade-off exists between learning ability of larvae and their hunting efficiency. Larvae with greater asymmetry learn better than those with less, but the latter are better able to sense vibrational signals used to detect prey and can capture prey more quickly. Both traits, learning ability and hunting efficiency, present obvious fitness advantages; the trade-off between them may explain why behavioural asymmetry, which presumably stems from brain lateralization, is relatively rare in natural antlion populations.

Larval antlions with more pronounced behavioural asymmetry show enhanced cognitive skills.

Brain lateralization is hypothesized to improve the efficiency of information processing. Here, we found that some Myrmeleon bore antlion larvae showed individual asymmetry in righting from a supine to normal position over one side of their body, which can be considered a reflection of greater brain lateralization. We demonstrated that these behaviourally asymmetrical individuals showed improved learning abilities, providing novel evidence that brain lateralization leads to beneficial effects on cognitive functions.

Sedentary antlion larvae (Neuroptera: Myrmeleontidae) use vibrational cues to modify their foraging strategies.

Learning abilities are exhibited by many animals, including insects. However, sedentary species are typically believed to have low capacities and requirements for learning. Despite this view, recent studies show that even such inconspicuous organisms as larval antlions, which employ an ambush predation strategy, are capable of learning, although their learning abilities are rather simple, i.e., limited to the association of vibrational cues with the arrival of prey. This study demonstrates, for the first time, that antlion larvae can use vibrational cues for complex modifications of their foraging strategies. Specifically, antlion larvae rapidly learn to differentiate between the vibrational cues associated with prey of different sizes, and they save resources by ignoring smaller prey in favour of larger, more energetically profitable prey. Moreover, antlion larvae can learn to associate vibrational cues with the loss of their prey, and they respond by burying their victims under the sand more often and more rapidly than do individuals with no opportunities to form such associations. These findings provide not only new insights into the cognitive abilities of animals but also support for the optimal foraging strategy concept, suggesting the importance of maximizing fitness output by balancing the costs and benefits of alternative foraging strategies.

Occasional and constant exposure to dietary ethanol shortens the lifespan of worker honey bees.

Honey bees (Apis mellifera) are one of the most crucial pollinators, providing vital ecosystem services. Their development and functioning depend on essential nutrients and substances found in the environment. While collecting nectar as a vital carbohydrate source, bees routinely encounter low doses of ethanol from yeast fermentation. Yet, the effects of repeated ethanol exposure on bees' survival and physiology remain poorly understood. Here, we investigate the impacts of constant and occasional consumption of food spiked with 1% ethanol on honey bee mortality and alcohol dehydrogenase (ADH) activity. This ethanol concentration might be tentatively judged close to that in natural conditions. We conducted an experiment in which bees were exposed to three types of long-term diets: constant sugar solution (control group that simulated conditions of no access to ethanol), sugar solution spiked with ethanol every third day (that simulated occasional, infrequent exposure to ethanol) and daily ethanol consumption (simulating constant, routine exposure to ethanol). The results revealed that both constant and occasional ethanol consumption increased the mortality of bees, but only after several days. These mortality rates rose with the frequency of ethanol intake. The ADH activity remained similar in bees from all groups. Our findings indicate that exposure of bees to ethanol carries harmful effects that accumulate over time. Further research is needed to pinpoint the exact ethanol doses ingested with food and exposure frequency in bees in natural conditions.

Behavioral differences between pit-building antlions and wormlions suggest limits to convergent evolution.

Antlions and wormlions are distantly related insect taxa, both digging pits in loose soil and ambushing arthropod prey. Their hunting method, which is rare in the animal kingdom, is a clear example of convergent evolution. There is little research directly comparing the 2 pit-building taxa. Using the same experimental platform to investigate how they respond to biotic and abiotic environmental factors enables an examination of their convergence and its limits. We investigated the response of antlions and wormlions to 3 factors common in their daily life: disturbance to the pits, prey arrival, and conspecific competitors. Although both increased the pit size following disturbance, wormlions increased it faster than antlions. Antlions responded to prey faster than wormlions, but wormlions improved their response time over days. The most diverging response was toward conspecifics. Whereas antlions relocated their pits fast in response to increasing conspecific density, wormlions never relocated. We suggest explanations for the behavioral differences between the taxa. Our results imply that despite the similar hunting method of the 2 taxa they may differ greatly in their behavior, which in turn might have consequences for their habitat use and population dynamics.

Small workers are more persistent when providing and requiring help in a monomorphic ant.

The common sand-dwelling Formica cinerea ants possess monomorphic workers, yet with considerable and easily identified size variation. Considering the importance of body size in polymorphic ants and other animals, we test whether size-dependent differences in behaviour occur in this species. We focus on the behaviour of large and small foragers in the context of rescue occurring between nestmates when one of them is entrapped and requires help. We show that workers of different sizes are characterized by a similar frequency of rescue activity and time delay to the first act of rescue. However, small workers rescue for longer than large workers. These results indicate that, although there is no size-related rescue specialization in F. cinerea foragers, small rescuers behave differently than large ones in terms of rescue persistence. Additionally, we show that small workers are more active when trapped. We suggest that variation in behavioural persistence of differently-sized workers may increase the efficiency of rescue actions. This study is the first to find a connection between body size and rescue behaviour in ants and the first to quantify and analyze the behaviour of individuals in need of rescue. These findings add substantially to our understanding of social insects and, more generally, highlight the need to study among-individual behavioural variation in social animals, including those in which body size is judged minute and irrelevant.

Heat stress during development makes antlion larvae more responsive to vibrational cues.

We investigated the effects of heat stress on the responsiveness to vibrational cues, our measure of perceptual ability, in Myrmeleon bore antlion larvae (Neuroptera: Myrmeleontidae). We reared these trap-building predatory larvae under 2 heat stress regimes (mild, 30°C, and harsh, 36°C), and after they progressed from one instar stage to another, we tested their perceptual ability in common unchallenging conditions. We hypothesized that exposure to the harsh heat stress regime would impose costs resulting in handicapped vibration responsiveness. We found that the harsh heat stress regime generated more stressful conditions for the larvae, as evidenced by increased mortality and postponed molting, and the loss of body mass among larger larvae. Furthermore, among the individuals who remained alive, those originating from the harsh heat stress regime were characterized by higher vibration responsiveness. Our results suggest 2 not mutually exclusive scenarios. Costly heat stress conditions can sieve out individuals characterized by poor perceptual ability or surviving individuals can attempt to hunt more efficiently to compensate for the physiological imbalance caused by heat stress. Both of these mechanisms fit into the ongoing debate over how adaptation and plasticity contribute to shaping insect communities exposed to heat stress.

Comparative analysis of experimental testing procedures for the elicitation of rescue actions in ants.

Rescue behavior is observed when 1 individual provides help to another individual in danger. Most reports of rescue behavior concern ants (Formicidae), in which workers rescue each other from various types of entrapment. Many of these entrapment situations can be simulated in the laboratory using an entrapment bioassay, in which ants confront a single endangered nest mate entrapped on a sandy arena by means of an artificial snare. Here, we compared numerous characteristics of rescue actions (contact between individuals, digging around the entrapped individual, pulling at its body parts, transport of the sand covering it, and biting the snare entrapping it) in Formica cinerea ants. We performed entrapment tests in the field and in the laboratory, with the latter under varying conditions in terms of the number of ants potentially engaged in rescue actions and the arena substrate (marked or unmarked by ants' pheromones). Rescue actions were more probable and pronounced in the field than in the laboratory, regardless of the type of test. Moreover, different test types in the laboratory yielded inconsistent results and showed noteworthy variability depending on the tested characteristic of rescue. Our results illustrate the specifics of ant rescue actions elicited in the natural setting, which is especially important considering the scarcity of field data. Furthermore, our results underline the challenges related to the comparison of results from different types of entrapment tests reported in the available literature. Additionally, our study shows how animal behavior differs in differing experimental setups used to answer the same questions.

Past thermal conditions affect hunting behaviour in larval antlions.

Some sit-and-wait predators, such as antlion larvae, construct traps to capture passing prey. The location of these traps depends on many abiotic and biotic factors, including temperature and the presence of conspecifics, which probably stimulate behaviours that minimize the costs and maximize the benefits of trap building. Here, we exposed second instar antlion larvae to elevated temperatures of 25°C (mild treatment) or 31°C (harsh treatment) for one month and then transferred them to common conditions (20°C) to examine the effects of previous thermal treatment on aggregation tendency and trap size. We predicted that antlions that experienced harsh conditions would subsequently increase the neighbouring distance and trap diameter to reduce competition with conspecifics and improve prey capture success, compensating for past conditions. In contrast with these predictions, antlions exposed to harsh conditions displayed a trend in the opposite direction, towards the decreased neighbouring distance. Furthermore, some of these antlions also built smaller traps. We discuss possible reasons for our results. The effects of previous thermal exposure have rarely been considered in terms of trap construction in antlions. Described effects may possibly apply to other sit-and-wait predators and are significant considering that many of these predators are long-lived.

"O Sister, Where Art Thou?"-A Review on Rescue of Imperiled Individuals in Ants.

Altruism is defined as an action that decreases the lifetime direct fitness of an actor and benefits one or more recipients. This phenomenon, which is generally difficult to understand and explain, requires special research attention. The subject of this review, rescue, is a type of altruistic behavior in which the actor puts itself at risk to save another individual, the recipient, that is in danger. The highest numbers of published empirical works have been devoted to rescue behavior in ants and they have enormous potential for further study. We review studies devoted to the subject and group them into four main areas of research on ant rescue actions: (1) variation in rescue behavior activity on a between-individual scale, (2) factors contributing to the evolution of rescue behavior on a between-species scale, (3) rescue behavior releaser signals and (4) rescue behavior benefits and costs. We highlight the progress in research on rescue behavior in ants, indicate that this behavior is probably much more common than previously thought yet thus far demonstrated in only a few species, and uncover research gaps and open questions that remain unexplored. We additionally point out some gaps in knowledge that become evident when research devoted to rescue behavior in rats, the second most studied group of animals in this context, is briefly overviewed. We hope to help navigate among studies on rescue behavior and provide the most up-to-date summary of the relevant literature. Moreover, we hope to encourage and facilitate researchers in behavioral ecology and other subdisciplines to further experimentally analyze rescue behavior, not only in ants but also in other taxa.

Heat wave effects on the behavior and life-history traits of sedentary antlions.

Research on the behavioral responses of animals to extreme weather events, such as heat wave, is lacking even though their frequency and intensity in nature are increasing. Here, we investigated the behavioral response to a simulated heat wave in two species of antlions (Neuroptera: Myrmeleontidae). These insects spend the majority of their lives as larvae and live in sandy areas suitable for a trap-building hunting strategy. We used larvae of Myrmeleon bore and Euroleon nostras, which are characterized by different microhabitat preferences-sunlit in the case of M. bore and shaded in the case of E. nostras. Larvae were exposed to fluctuating temperatures (40 °C for 10 h daily and 25 °C for the remaining time) or a constant temperature (25 °C) for an entire week. We found increased mortality of larvae under heat. We detected a reduction in the hunting activity of larvae under heat, which corresponded to changes in the body mass of individuals. Furthermore, we found long-term consequences of the simulated heat wave, as it prolonged the time larvae needed to molt. These effects were pronounced in the case of E. nostras but did not occur or were less pronounced in the case of M. bore, suggesting that microhabitat-specific selective pressures dictate how well antlions handle heat waves. We, thus, present results demonstrating the connection between behavior and the subsequent changes to fitness-relevant traits in the context of a simulated heat wave. These results illustrate how even closely related species may react differently to the same event.

Honeybee rebel workers invest less in risky foraging than normal workers.

In eusocial insect colonies, workers have individual preferences for performing particular tasks. Previous research suggests that these preferences might be associated with worker reproductive potential; however, different studies have yielded inconsistent results. This study constitutes the first comparison of foraging preferences between genetically similar normal and rebel honeybee workers, which present different reproductive potential. We found that rebels, which have a higher reproductive potential than normal workers, displayed a delayed onset of foraging and a stronger tendency to collect nectar compared with normal workers. These results support the hypothesis that workers with high reproductive potential invest more in their own egg laying and avoid risky tasks such as foraging. In contrast, the results do not support the hypothesis that reproductive workers initiate foraging earlier in life than normal workers and specialize in pollen foraging.

Thermal dependence of trap building in predatory antlion larvae (Neuroptera: Myrmeleontidae).

Trap-building predators remain under strong selection from thermal microenvironments. To address how soil temperature and body size affect trap building, we conducted a laboratory experiment using larvae of the antlion Myrmeleon bore at six ecologically relevant temperatures. Larger larvae built larger traps, and warmer soil led to more and larger traps. Body mass did not alter the dependence of trap building on temperature. Our results suggest that the physiological capacity of antlion larvae, which is affected by larval size and body temperature, is the major determinant of trap building. This effect should be considered when assessing interactions between antlions and prey.

Honeybees show adaptive reactions to ethanol exposure.

The honeybee is being developed as a simple invertebrate model for alcohol-related studies. To date, several effects of ethanol consumption have been demonstrated in honeybees, but the tolerance effect, one of the hallmarks of alcohol overuse, has never been shown. Here, we confirm our hypothesis that the response to ethanol (in terms of motor impairment) is lower in bees that have previously experienced intoxication than in bees encountering ethanol for the first time, indicating that the chronic tolerance effect occurs in honeybees. Furthermore, we investigated the basis of this effect and found that it likely results from conditioned compensatory responses to cues associated with ethanol delivery. Our findings significantly improve our understanding of the suitability of honeybees as models for alcoholism-related research and underline the first and foremost function of all conditioned reactions - their adaptive value.

Rebel honeybee workers have a tendency to become intraspecific reproductive parasites.

Worker honeybees may reproduce in either their own or foreign colonies; the latter situation is termed intraspecific reproductive parasitism (IRP). In this study, we compared the tendency for IRP between normal honeybee workers, which are characterized by a relatively low reproductive potential, and "rebel workers", a recently discovered subcaste of honeybee workers characterized by a high reproductive potential that develops when the colony is without a queen. We expected that the high reproductive potential of the rebel workers would influence their reproductive strategy and that these individuals would drift to other colonies to lay eggs more often than normal workers. The results confirm our expectations and show that rebel workers are more likely than normal workers to drift to foreign colonies. The rebel workers also preferred to drift to queenless colonies than to queenright colonies, while the normal workers did not show this preference. This study indicates that rebel workers have a tendency for IRP, which may be responsible for the maintenance of the rebel worker strategy in bee populations.

Pro-social behaviour of ants depends on their ecological niche-Rescue actions in species from tropical and temperate regions.

Some ants display rescue behaviour, which is performed by nearby nestmates and directed at individuals in danger. Here, using several ant species, we demonstrate that rescue behaviour expression matches predicted occurrences based on certain aspects of species' ecological niches. Rescue occurred in sand-dwelling ants exposed both to co-occurring antlion larvae, representing the threat of being captured by a predator, and to nest cave-ins, representing the threat of being trapped in a collapsed nest chamber. Rescue also occurred in forest groundcover ants exposed to certain entrapment situations. However, rescue never occurred in species associated with open plains, which nest in hardened soils and forage largely on herbaceous plants, or in ants living in close mutualistic relationships with their host plants. In addition, because we tested each species in two types of tests, antlion larva capture tests and artificial entrapment tests, we highlight the importance of accounting for test context in studying rescue behaviour expression.

Increased Risk Proneness or Social Withdrawal? The Effects of Shortened Life Expectancy on the Expression of Rescue Behavior in Workers of the ant Formica cinerea (Hymenoptera: Formicidae).

In social insects behavioral consequences of shortened life expectancy include, among others, increased risk proneness and social withdrawal. We investigated the impact of experimental shortening of life expectancy of foragers of the ant Formica cinerea achieved by their exposure to carbon dioxide on the expression of rescue behavior, risky pro-social behavior, tested by means of two bioassays during which a single worker (rescuer) was confronted with a nestmate (victim) attacked by a predator (antlion larva capture bioassay) or immobilized by an artificial snare (entrapment bioassay). Efficacy of carbon dioxide poisoning in shortening life expectancy was confirmed by the analysis of ant mortality. Rescue behavior observed during behavioral tests involved digging around the victim, transport of the sand covering the victim, pulling the limbs/antennae/mandibles of the victim, direct attack on the antlion (in antlion larva capture tests), and snare biting (in entrapment tests). The rate of occurrence of rescue behavior was lower in ants with shortened life expectancy, but that effect was significant only in the case of the entrapment bioassay. Similarly, only in the case of the entrapment bioassay ants with shortened life expectancy displayed rescue behavior after a longer latency and devoted less time to that behavior than ants from the control groups. Our results demonstrated that in ant workers shortened life expectancy may lead to reduced propensity for rescue behavior, most probably as an element of the social withdrawal syndrome that had already been described in several studies on behavior of moribund ants and honeybees.