Daily Thermal Fluctuations Experienced by Pupae via Rhythmic Nursing Behavior Increase Numbers of Mushroom Body Microglomeruli in the Adult Ant Brain.

Social insects control brood development by using different thermoregulatory strategies. Camponotus mus ants expose their brood to daily temperature fluctuations by translocating them inside the nest following a circadian rhythm of thermal preferences. At the middle of the photophase brood is moved to locations at 30.8°C; 8 h later, during the night, the brood is transferred back to locations at 27.5°C. We investigated whether daily thermal fluctuations experienced by developing pupae affect the neuroarchitecture in the adult brain, in particular in sensory input regions of the mushroom bodies (MB calyces). The complexity of synaptic microcircuits was estimated by quantifying MB-calyx volumes together with densities of presynaptic boutons of microglomeruli (MG) in the olfactory lip and visual collar regions. We compared young adult workers that were reared either under controlled daily thermal fluctuations of different amplitudes, or at different constant temperatures. Thermal regimes significantly affected the large (non-dense) olfactory lip region of the adult MB calyx, while changes in the dense lip and the visual collar were less evident. Thermal fluctuations mimicking the amplitudes of natural temperature fluctuations via circadian rhythmic translocation of pupae by nurses (amplitude 3.3°C) lead to higher numbers of MG in the MB calyces compared to those in pupae reared at smaller or larger thermal amplitudes (0.0, 1.5, 9.6°C), or at constant temperatures (25.4, 35.0°C). We conclude that rhythmic control of brood temperature by nursing ants optimizes brain development by increasing MG densities and numbers in specific brain areas. Resulting differences in synaptic microcircuits are expected to affect sensory processing and learning abilities in adult ants, and may also promote interindividual behavioral variability within colonies.

Long-term avoidance memory formation is associated with a transient increase in mushroom body synaptic complexes in leaf-cutting ants.

Long-term behavioral changes related to learning and experience have been shown to be associated with structural remodeling in the brain. Leaf-cutting ants learn to avoid previously preferred plants after they have proved harmful for their symbiotic fungus, a process that involves long-term olfactory memory. We studied the dynamics of brain microarchitectural changes after long-term olfactory memory formation following avoidance learning in Acromyrmex ambiguus. After performing experiments to control for possible neuronal changes related to age and body size, we quantified synaptic complexes (microglomeruli, MG) in olfactory regions of the mushroom bodies (MBs) at different times after learning. Long-term avoidance memory formation was associated with a transient change in MG densities. Two days after learning, MG density was higher than before learning. At days 4 and 15 after learning-when ants still showed plant avoidance-MG densities had decreased to the initial state. The structural reorganization of MG triggered by long-term avoidance memory formation clearly differed from changes promoted by pure exposure to and collection of novel plants with distinct odors. Sensory exposure by the simultaneous collection of several, instead of one, non-harmful plant species resulted in a decrease in MG densities in the olfactory lip. We hypothesize that while sensory exposure leads to MG pruning in the MB olfactory lip, the formation of long-term avoidance memory involves an initial growth of new MG followed by subsequent pruning.

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.

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.

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.