Ants evade harmful food by active abandonment.

Invasive ants, such as the Argentine ant, pose a severe economic and ecological threat. Despite advancements in baiting techniques, effectively managing established ant populations remains a daunting challenge, often ending in failure. Ant colonies employ behavioural immunity against pathogens, raising the question of whether ants can collectively respond to toxic baits. This study investigates whether ant colonies actively abandon palatable but harmful food sources. We provided two sucrose feeders, each generating a new foraging trail, with one transitioning to offering toxic food. Six hours later, ant activity on that path decreases, while activity on the non-toxic food and the trunk trail remains unaffected, excluding factors like population decline or satiation as reasons for the activity decline. Laboratory experiments confirmed that ants remained alive six hours after ingesting toxic food. Ant presence remains low on the toxic food path for days, gradually decreasing along the nearest section of the trunk trail. This abandonment behaviour minimises the entry of harmful food into the nest, acting as a protective social mechanism. The evasion of toxic bait-treated areas likely contributes considerably to control failures. Understanding the behavioural response to toxic baits is essential for developing effective strategies to combat invasive ant species.

Inhibition of serotonergic signaling induces higher consumption of both sucrose solution and toxic baits in carpenter ants.

Biogenic amines play an important role in the regulation of appetitive responses in insects. Among them, serotonin (5-HT) regulates feeding-related processes in numerous insect species. In carpenter ants, 5-HT administration has been shown to depress feeding behavior, thus opening the possibility of using 5-HT modulation in control strategies against those species considered as pest. Here we studied if administration of a 5-HT antagonist, ketanserin, promotes feeding of a sucrose solution and a toxic bait in carpenter ants Camponotus mus. We found that 3 h after a single oral administration of ketanserin, the mass of sucrose solution consumed by carpenter ants increased significantly. A similar effect was found after a chronic administration that lasted 5 days. Yet, ketanserin did neither affect the intake rates nor the activity of the pharyngeal pump that mediates feeding dynamics. In addition, ketanserin promoted the consumption of a toxic bait based on boric acid. Our results thus show that feeding motivation and consumption of both sucrose solution and a toxic bait can be enhanced via prior administration of ketanserin. We discuss the possible mechanisms underlying these effects and conclude that understanding basic physiological and neural principles that underlie feeding motivation allows establishing more efficient control strategies for pest insects.

Laboratory and field insights into the dynamics and behavior of Argentine ants, Linepithema humile, feeding from hydrogels.

BACKGROUND: Hydrogels that have absorbed a liquid containing a toxicant are a novel form of bait-delivery for ant control. Here, we study the abilities of Argentine ant (Linepithema humile) workers to imbibe liquid from hydrogels. We quantified feeding behavior with: (i) hydrogels containing different sucrose concentrations (20, 30, 40 and 50% w/w); (ii) hydrogels versus liquid droplets; and (iii) hydrogel age (air exposure time). We also performed a field assay to assess visits by L. humile and other ant species to hydrogels. RESULTS: Ingested volume and feeding time decreased with increasing sucrose concentrations, but the number and duration of pauses were similar. Feeding from hydrogels was slower than from a liquid droplet and ants imbibed less liquid and fed for shorter times from hydrogels. Feeding time increased with hydrogel age, whereas ingested volume decreased and approached zero after 120 min under laboratory conditions. In the field, ants attended the hydrogels during the full 120-min study period. When L. humile workers found a hydrogel, they monopolized it to the exclusion of other ant species. L. humile occupied and dominated hydrogels predominantly in shaded locations. CONCLUSION: Hydrogels with sucrose concentrations no greater than 30% appear best for liquid uptake by L. humile. Hydrogels not in direct sunlight will have greater attendance by L. humile and, therefore, less attendance by non-target ant species. Shady and humid places may prolong the longevity of hydrogels, which would imply higher intakes. © 2021 Society of Chemical Industry.

Viscosity as a key factor in decision making of nectar feeding ants.

It is well known that viscosity reduces the intake rates in nectar-feeding insects, such as nectivorous ants, though it remains unclear whether viscosity imposes a higher energy investment in these insects, and how this affects their feeding motivation. To address this issue, we studied feeding behavior, metabolism, and pharyngeal pump activity in the carpenter ant Camponotus mus during ingestion of ad libitum sucrose solutions. In some solutions tylose was added to modify viscosity without changing its sucrose concentration, in a way that allowed comparing: (1) two solutions with the same viscosity and different sucrose concentration (10 T and 50), and (2) two solutions with different viscosity and the same sucrose concentration (50 and 50 T). The viscosity increase was detrimental to the metabolic rate and energy balance. Ants feeding on a solution with high sucrose concentration and increased viscosity (50 T) spent extra-time until reaching a crop load similar to that reached by ingesting the solution without tylose (50). For all solutions offered, ants started feeding with the same pharyngeal pump frequencies, reflecting a similar motivation. Interesting, when ants fed on a low sucrose concentration and increased viscosity solution (10 T), their pump frequencies dropped rapidly respect to the pure-sucrose solution (50). On the contrary, pump frequencies for 50 and 50 T remained similar until the end of the intake. Since the pump frequency is strongly modulated by the ant motivation, an increase in viscosity with low sucrose content, demotivates the ants rapidly, suggesting a rapid integration of different kinds of information about the food value. Our results helped to understand how nectivorous ants could modulate their foraging decision-making.

Trail pheromone modulates subjective reward evaluation in Argentine ants.

The Argentine ant, Linepithema humile, is native to South America but has become one of the most invasive species in the world. These ants heavily rely on trail pheromones for foraging, and previous studies have focused on such signals to develop a strategy for chemical control. Here, we studied the effects of pre-exposure to the trail pheromone on sugar acceptance and olfactory learning in Argentine ants. We used the synthetic trail pheromone component (Z)-9-hexadecenal, which triggers the same attraction and trail-following behavior as the natural trail pheromone. We found that pre-exposure to (Z)-9-hexadecenal increases the acceptance of sucrose solutions of different concentrations, thus changing the ants' subjective evaluation of a food reward. However, although ants learned to associate an odor with a sucrose reward, pheromone pre-exposure affected neither the learning nor the mid-term memory of the odor-reward association. Taking into account the importance of the Argentine ant as a pest and invasive organism, our results highlight the importance of pheromonal cues in resource evaluation, a fact that could be useful in control strategies implemented for this species.

Environmental temperature affects the dynamics of ingestion in the nectivorous ant Camponotus mus.

Environmental temperature influences physiology and behavior in animals in general and is particularly determinant in ectotherms. Not least because temperature defines metabolism and body temperature, muscle activity in insects also strongly depends on this factor. Here, we analyzed how environmental temperature influences the dynamics of ingestion due to its effect on the sucking pump muscles in the nectivorous ants Camponotus mus. Feeding behavior and sucking pump activity during sucrose solution ingestion were first recorded in a natural environment in an urban setting throughout the day and in different seasons. Then, controlled temperature experiments were performed in the laboratory. In both situations, feeding time decreased and pumping frequency increased with temperature. However, different pumping frequencies under a same temperature were also observed in different seasons. Besides, in the laboratory, the volume of solution ingested increased with temperature. Consequently, intake rate increased when temperature rose. This change was exclusively promoted by a variation in the pumping frequency while volume taken in per pump contraction was not affected by temperature. In summary, environmental temperature modified the dynamics of ingestion and feeding behavior by directly affecting pumping frequency.

Knowing the enemy: ant behavior and control in a pediatric hospital of Buenos Aires.

Ant control is difficult in systems even where a variety of control strategies and compounds are allowed; in sensitive places such as hospitals, where there are often restrictions on the methods and toxicants to be applied, the challenge is even greater. Here we report the methods and results of how we faced this challenge of controlling ants in a pediatric hospital using baits. Our strategy was based on identifying the species present and analyzing their behavior. On the one hand, we evaluated outdoors in the green areas of the hospital, the relative abundance of ant genera, their food preferences and the behavioral dominances. On the other hand, control treatments were performed using separately two boron compounds added to sucrose solution which was not highly concentrated to avoid constrains due to the viscosity. Most of the species in the food preference test accepted sugary food; only one species was recorded to visit it less than the protein foods. This result was consistent with the efficacy of control treatments by sugary baits within the rooms. For species that showed good acceptance of sugar solutions in the preference test outdoors, sugar bait control indoors was 100& effective. Conversely, for the only species that foraged significantly less on sugar food, the bait treatment was ineffective. This work reveals the importance of considering the behavior and feeding preferences of the species to be controlled by toxic baits.

Efficient egress of escaping ants stressed with temperature.

In the present work we investigate the egress times of a group of Argentine ants (Linepithema humile) stressed with different heating speeds. We found that the higher the temperature ramp is, the faster ants evacuate showing, in this sense, a group-efficient evacuation strategy. It is important to note that even when the life of ants was in danger, jamming and clogging was not observed near the exit, in accordance with other experiments reported in the literature using citronella as aversive stimuli. Because of this clear difference between ants and humans, we recommend the use of some other animal models for studying competitive egress dynamics as a more accurate approach to understanding competitive egress in human systems.

Asymmetrical behavioral response towards two boron toxicants depends on the ant species (Hymenoptera: Formicidae).

Urban ants are a worldwide critical household pests, and efforts to control them usually involve the use of alimentary baits containing slow-acting insecticides. A common toxicant used is boron, either as borax or boric acid. However, the presence of these compounds can affect the consumption of baits by reducing their acceptance and ingestion. Moreover, as feeding motivation varies widely, according not only to food properties but also to colony conditions, bait consumption might be diminished further in certain situations. In this study, we compared the feeding response of ants toward two boron toxic baits (boric acid and borax) in low motivation situations that enhance any possible phago-deterrence the baits may produce. Most studies investigating bait ingestion evaluate whole nests or groups of ants; here, we analyzed the individual ingestion behavior and mortality of the Argentine ant, Linepithema humile (Mayr), and the carpenter ant, Camponotus mus (Roger), for two boron baits, to detect which compound generates a higher rejection in each of these species. Although these two species have similar feeding habits, our results showed that ants under low motivation conditions reduced the acceptance and consumption of the toxic baits asymmetrically. While L. humile mostly rejected the borax, C. mus rejected the boric acid. These results denote the importance of considering the preference of each species when developing a pest management strategy.

Serotonin depresses feeding behaviour in ants.

Feeding behaviour is a complex functional system that relies on external signals and the physiological state of the animal. This is also the case in ants as they vary their feeding behaviour according to food characteristics, environmental conditions and - as they are social insects - to the colony's requirements. The biogenic amine serotonin (5-HT) was shown to be involved in the control and modulation of many actions and processes related to feeding in both vertebrates and invertebrates. In this study, we investigated whether 5-HT affects nectar feeding in ants by analysing its effect on the sucking-pump activity. Furthermore, we studied 5-HT association with tissues and neuronal ganglia involved in feeding regulation. Our results show that 5-HT promotes a dose-dependent depression of sucrose feeding in Camponotus mus ants. Orally administered 5-HT diminished the intake rate by mainly decreasing the volume of solution taken per pump contraction, without modifying the sucrose acceptance threshold. Immunohistochemical studies all along the alimentary canal revealed 5-HT-like immunoreactive processes on the foregut (oesophagus, crop and proventriculus), while the midgut and hindgut lacked 5-HT innervation. Although the frontal and suboesophageal ganglia contained 5-HT immunoreactive cell bodies, serotonergic innervation in the sucking-pump muscles was absent. The results are discussed in the frame of a role of 5-HT in feeding control in ants.

Calcium imaging in the ant Camponotus fellah reveals a conserved odour-similarity space in insects and mammals.

BACKGROUND: Olfactory systems create representations of the chemical world in the animal brain. Recordings of odour-evoked activity in the primary olfactory centres of vertebrates and insects have suggested similar rules for odour processing, in particular through spatial organization of chemical information in their functional units, the glomeruli. Similarity between odour representations can be extracted from across-glomerulus patterns in a wide range of species, from insects to vertebrates, but comparison of odour similarity in such diverse taxa has not been addressed. In the present study, we asked how 11 aliphatic odorants previously tested in honeybees and rats are represented in the antennal lobe of the ant Camponotus fellah, a social insect that relies on olfaction for food search and social communication. RESULTS: Using calcium imaging of specifically-stained second-order neurons, we show that these odours induce specific activity patterns in the ant antennal lobe. Using multidimensional analysis, we show that clustering of odours is similar in ants, bees and rats. Moreover, odour similarity is highly correlated in all three species. CONCLUSION: This suggests the existence of similar coding rules in the neural olfactory spaces of species among which evolutionary divergence happened hundreds of million years ago.

Olfactory memory established during trophallaxis affects food search behaviour in ants.

Camponotus mus ants can associate sucrose and odour at the source during successive foraging cycles and use this memory to locate the nectar in the absence of other cues. These ants perform conspicuous trophallactic behaviour during recruitment while foraging for nectar. In this work, we studied whether Camponotus mus ants are able to establish this odour-sucrose association in the social context of trophallaxis and we evaluated this memory in another context previously experienced by the ant, as a nectar source. After a single trophallaxis of a scented solution, the receiver ant was tested in a Y-maze without any reward, where two scents were presented: in one arm, the solution scent and in the other, a new scent. Ants consistently chose the arm with the solution scent and stayed longer therein. Trophallaxis duration had no effect on the arm choice or with the time spent in each arm. Workers are able to associate an odour (conditioned stimulus) with the sucrose (unconditioned stimulus) they receive through a social interaction and use this memory as choice criteria during food searching.

Differential conditioning and long-term olfactory memory in individual Camponotus fellah ants.

Individual Camponotus fellah ants perceive and learn odours in a Y-maze in which one odour is paired with sugar (CS+) while a different odour (CS-) is paired with quinine (differential conditioning). We studied olfactory retention in C. fellah to determine whether olfactory learning leads to long-term memory retrievable 24 h and 72 h after training. One and 3 days after training, ants exhibited robust olfactory memory through a series of five successive retention tests in which they preferred the CS+ and stayed longer in the arm presenting it. In order to determine the nature of the associations memorized, we asked whether choices within the Y-maze were driven by excitatory memory based on choosing the CS+ and/or inhibitory memory based on avoiding the CS-. By confronting ants with a novel odour vs either the CS+ or the CS- we found that learning led to the formation of excitatory memory driving the choice of the CS+ but no inhibitory memory based on the CS- was apparent. Ants even preferred the CS- to the novel odour, thus suggesting that they used the CS- as a contextual cue in which the CS+ was embedded, or as a second-order cue predicting the CS+ and thus the sugar reward. Our results constitute the first controlled account of olfactory long-term memory in individual ants for which the nature of associations could be precisely characterized.

Sucking pump activity in feeding behaviour regulation in carpenter ants.

Modulation of liquid feeding-rate would allow insects to ingest more food in the same time when this was required. Ants can vary nectar intake rate by increasing sucking pump frequency according to colony requirements. We analysed electrical signals generated by sucking pump activity of ants during drinking solutions of different sucrose concentrations and under different carbohydrate-deprivation levels. Our aim was to define parameters that characterize the recordings and analyse their relationship with feeding behaviour. Signals showed that the initial and final frequencies of sucking pump activity, as well as the difference between them were higher in sugar-deprived ants. However, these parameters were not influenced by sucrose solution concentration, which affected the number of pump contractions and the volume per contraction. Unexpectedly, we found two different responses in feeding behaviour of starved and non-starved ants depending on concentration. Starved ants drank dilute solutions for the same length of time as non-starved ants but ingested higher volumes. While drinking the concentrated solutions, starved ants drank the same volume, but did so in a shorter time than the non-starved ones. Despite these differences, for each analysed concentration the total number of pump contractions remained constant independently of sugar-deprivation level. These results are discussed in the frame of feeding regulation and decision making in ant foraging behaviour.

Nectar intake rate is modulated by changes in sucking pump activity according to colony starvation in carpenter ants.

Dynamics of fluid feeding has been deeply studied in insects. However, the ability to vary the nectar-intake rate depending only on the carbohydrate deprivation has been clearly demonstrated only in Camponotus mus ants. When insect morphometry and fluid properties remain constant, changes in intake rate could only be attributed to variations in sucking pump activity. Previous records of the electrical activity generated during feeding in C. mus have revealed two different signal patterns: the regular (RP, frequencies: 2-5 Hz) and the irregular (IP, frequencies: 7-12 Hz). This work studies the mechanism underlying food intake-rate modulation in ants by analysing whether these patterns are involved. Behaviour and electrical activity generated by ants at different starvation levels were analysed during feeding on sucrose solutions. Ants were able to modulate the intake rate for a variety of sucrose concentrations (10, 40 and 60%w/w). The IP only occurred for 60% of solutions and its presence did not affect the intake rate. However, during the RP generated under the starved state, we found frequencies up to 7.5 Hz. RP frequencies positively correlated with the intake-rate for all sucrose concentrations. Hence, intake-rate modulation according to sugar deprivation is mainly achieved by the ant's ability to vary the pumping frequency.

Electrical signals during nectar sucking in the carpenter ant Camponotus mus.

Ants of the same size can vary their intake rate of a given sucrose solution depending on the colony's needs for carbohydrates. As this capacity has not yet been described for another insect, the question of how they can do that was the focus of our work. When viscosity and ant-morphometry remain constant, changes in intake rate can only be attributed to the sucking forces. The aim of this study was to analyze the nectar sucking activity in the ant Camponotus mus. Feeding behavior seems to be under motivational control; therefore, we developed a non-invasive experimental device. We recorded the electrical signal generated during nectar feeding by offering ants sucrose solutions of different concentrations (from 10%w/w to 70%w/w). The signal frequency was between 2 and 12 peaks/s. We could distinguish two different patterns of electrical signal during feeding depending on the solution concentration. Only the more concentrated solutions reached frequencies higher than 7 peaks/s and the signal performance was quite irregular. For the other concentrations (10%, 30% and 50%), signal frequencies were lower than 6 peaks/s and the signal pattern was sinusoidal, regular and decreased with intake in all cases. We discuss the possible implications of these two signal patterns.

Nectar foraging behaviour is affected by ant body size in Camponotus mus.

The nectivorous ant Camponotus mus shows a broad size variation within the worker caste. Large ants can ingest faster and larger loads than small ones. Differences in physiological abilities in fluid ingestion due to the insect size could be related to differences in decision-making according to ant size during nectar foraging. Sucrose solutions of different levels of sugar concentration (30% or 60%w/w), viscosity (high or low) or flow rate (ad libitum or 1microl/min) were offered in combination to analyse the behavioural responses to each of these properties separately. Differences were found depending on ant body size and the property compared. A regulated flow produced smaller crop loads for medium and large ants compared to the same solution given ad libitum. All foragers remained longer times feeding at the regulated flow source but larger ants often made longer interruptions. When sugar concentration was constant but viscosity was high, only large ants increased feeding time. Constant viscosity with different sugar concentration determined longer feeding time and bigger loads for the most concentrated solution for small but not for large ants. Small ants reached similar crop loads in a variety of conditions while large ants did not. These differences could be evidence of a possible specialization for nectar foraging based on ant body size.