Gigapixel behavioral and neural activity imaging with a novel multi-camera array microscope

The dynamics of living organisms are organized across many spatial scales, yet existing, cost-effective imaging systems can measure only a subset of these scales at once. Here, we have created a scalable multi-camera array microscope (MCAM) that enables comprehensive high-resolution recording from multiple spatial scales simultaneously, ranging from cellular structures to large-group behavioral dynamics. By collecting data from up to 96 cameras, we computationally generate gigapixel-scale images and movies near cellular resolution and 5 um sensitivity over hundreds of square centimeters. This allows us to observe the behavior and fine anatomical features of numerous freely moving model organisms on multiple spatial scales, including larval zebrafish, fruit flies, nematodes, carpenter ants, and slime mold. The MCAM architecture allows stereoscopic tracking of the z-position of organisms using the overlapping field of view from adjacent cameras. Further, we demonstrate the ability to acquire dual color fluorescence video of multiple freely moving zebrafish, recording neural activity via ratiometric calcium imaging. Overall, the MCAM provides a powerful platform for investigating cellular and behavioral processes across a wide range of spatial scales, but without the bottlenecks imposed by single-camera image acquisition systems.

Formic acid modulates latency and accuracy of nestmate recognition in carpenter ants

Decision-making processes face the dilemma of being accurate or faster, a phenomenon that has been described as speed-accuracy trade-off in numerous studies on animal behaviour. In social insects, discriminating between colony members and aliens is subjected to this trade-off as rapid and accurate rejection of enemies is of primary importance for the maintenance and ecological success of insect societies. Recognition cues distinguishing aliens from nestmates are embedded in the cuticular hydrocarbon (CHC) layer and vary among colonies. In walking carpenter ants, exposure to formic acid (FA), an alarm pheromone, improves accuracy of nestmate recognition by decreasing both alien acceptance and nestmate rejection. Here we studied the effect of FA exposure on the spontaneous aggressive mandible opening response of harnessed Camponotus aethiops ants presented with either nestmate or alien CHCs. FA modulated both MOR accuracy and the latency to respond to odours of conspecifics. In particular, FA decreased MOR towards nestmates but increased it towards aliens. Furthermore, FA decreased MOR latency towards aliens but not towards nestmates. As response latency can be used as a proxy of response speed, we conclude that contrary to the prediction of the speed-accuracy trade-off theory, ants did not trade off speed against accuracy in the process of nestmate recognition.

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.

Antenna Cleaning Is Essential for Precise Behavioral Response to Alarm Pheromone and Nestmate–Non-Nestmate Discrimination in Japanese Carpenter Ants (Camponotus japonicus)

Self-grooming of the antennae is frequently observed in ants. This antennal maintenance behavior is presumed to be essential for effective chemical communication but, to our knowledge, this has not yet been well studied. When we removed the antenna-cleaning apparatuses of the Japanese carpenter ant (C. japonicus) to limit the self-grooming of the antennae, the worker ants demonstrated the self-grooming gesture as usual, but the antennal surface could not be sufficiently cleaned. By using scanning electron microscopy with NanoSuit, we observed the ants’ antennae for up to 48 h and found that the antennal surfaces gradually became covered with self-secreted surface material. Concurrently, the self-grooming-limited workers gradually lost their behavioral responsiveness to undecane—the alarm pheromone. Indeed, their locomotive response to the alarm pheromone diminished for up to 24 h after the antenna cleaner removal operation. In addition, the self-grooming-limited workers exhibited less frequent aggressive behavior toward non-nestmate workers, and 36 h after the operation, approximately half of the encountered non-nestmate workers were accepted as nestmates. These results suggest that the antennal sensing system is affected by excess surface material; hence, their proper function is prevented until they are cleaned.

All sugars ain't sweet: selection of particular mono-, di- and trisaccharides by western carpenter ants and European fire ants

Ants select sustained carbohydrate resources, such as aphid honeydew, based on many factors including sugar type, volume and concentration. We tested the hypotheses (H1–H3) that western carpenter ants, Camponotus modoc, seek honeydew excretions from Cinara splendens aphids based solely on the presence of sugar constituents (H1), prefer sugar solutions containing aphid-specific sugars (H2) and preferentially seek sugar solutions with higher sugar content (H3). We further tested the hypothesis (H4) that workers of both Ca. modoc and European fire ants, Myrmica rubra , selectively consume particular mono-, di- and trisaccharides. In choice bioassays with entire ant colonies, sugar constituents in honeydew (but not aphid-specific sugar) as well as sugar concentration affected foraging decisions by Ca. modoc . Both Ca. modoc and M. rubra foragers preferred fructose to other monosaccharides (xylose, glucose) and sucrose to other disaccharides (maltose, melibiose, trehalose). Conversely, when offered a choice between the aphid-specific trisaccharides raffinose and melezitose, Ca. modoc and M. rubra favoured raffinose and melezitose, respectively. Testing the favourite mono-, di- and trisaccharide head-to-head, both ant species favoured sucrose. While both sugar type and sugar concentration are the ultimate cause for consumption by foraging ants, strong recruitment of nest-mates to superior sources is probably the major proximate cause.

Cooperative escape in ants and robots

The solution of complex problems by the collective action of simple agents in both biologically evolved and synthetically engineered systems involves cooperative action. Understanding the resulting emergent solutions requires integrating across the organismal behaviors of many individuals. Here we investigate an ecologically relevant collective task in black carpenter ants Camponotus pennsylvanicus: escape from a soft, erodible confining corral. Individual ants show a transition from individual exploratory excavation at random locations to spatially localized collective exploitative excavation and escape from the corral. A minimal continuum theory that coarse-grains over individual actions and considers their integrated influence on the environment leads to the emergence of an effective phase space of behaviors in terms of excavation strength and cooperation intensity. To test the theory over the range of predicted behaviors, we used custom-built robots (RAnts) that respond to stimuli and show the emergence (and failure) of cooperative excavation and escape. Overall, our approach shows how the cooperative completion of tasks can arise from relatively simple rules that involve the interaction of simple agents with a dynamically changing environment that serves as an enabler and modulator of behavior.

Natural history of two carpenter ants, Camponotus renggeri and C. rufipes (Hymenoptera: Formicidae), in an Atlantic Forest reserve

Widespread species face a wide variety of environmental challenges and their morphology, behavior, and natural history may change across their range. However, not rarely, natural history research is restricted to one or few locations. That is the case for Camponotus renggeri and C. rufipes. Both species occur across South America in different ecosystems, but most research on these species is restricted to the Brazilian savanna, known as Cerrado. Here, we describe the home range, nesting habits, and activity schedule of C. renggeri and C. rufipes in an Atlantic Forest reserve in SE Brazil. C. renggeri foraged exclusively during nighttime and C. rufipes remained active throughout the nycthmeron, but with little intensity during daylight hours. Most nests of both species were composed of dry straw, and home ranges varied from nearly 0.91 m2 (C. renggeri) to 1.79 mm2 (C. rufipes). Foraging areas overlapped, especially in C. renggeri. Our field study reinforces the importance of natural history and what it adds to our knowledge on the ecology and behavior of C. renggeri and C. rufipes in Atlantic Forest.

Know your foe: synanthropic spiders are deterred by semiochemicals of European fire ants

Many ants prey on spiders, suggesting that web-building spiders may avoid micro-locations near ant colonies or frequented by foraging ants. Here we tested the hypothesis that ant-derived semiochemicals deter synanthropic spiders. To generate stimuli, we exposed filter paper for 12 h to workers of European fire ants, Myrmica rubra, black garden ants, Lasius niger, or western carpenter ants, Camponotus modoc , and then offered select urban spiders in three-chamber olfactometer bioassays a choice between ant-exposed filter paper and unexposed control filter paper. Semiochemical deposits of M. rubra , but not of L. niger or C. modoc , had a significant deterrent effect on subadults of the false black widow, Steatoda grossa, the black widow, Latrodectus hesperus , and the hobo spider, Eratigena agrestis, as well as a moderate (but statistically not significant) deterrent effect on the cross spider, Araneus diadematus . The deterrent effect caused by semiochemical deposits of M. rubra may be attributable to the aggressive nature and efficient foraging of M. rubra in its invaded North American range, exerting selection pressure on community members to recognize M. rubra semiochemicals and to avoid micro-locations occupied by M. rubra .

Formic acid modulates latency and accuracy of nestmate recognition in carpenter ants

Decision-making processes face the dilemma of being accurate or faster, a phenomenon that has been described as speed-accuracy trade-off (SAT) in numerous studies on animal behaviour. In social insects, discriminating between colony members and aliens is subjected to this trade-off as rapid and accurate rejection of enemies is of primary importance for the maintenance and ecological success of insect societies. Recognition cues distinguishing aliens from nestmates are embedded in the cuticular hydrocarbon (CHC) layer and vary among colonies. In walking carpenter ants, exposure to formic acid (FA), an alarm pheromone, improves accuracy of nestmate recognition by decreasing both alien acceptance and nestmate rejection. Here we studied the effect of FA exposure on the spontaneous aggressive mandible opening response of harnessed Camponotus aethiops ants presented with either nestmate or alien CHCs. FA modulated both MOR accuracy and the latency to respond to odours of conspecifics. In particular, FA decreased MOR towards nestmates but increased it towards aliens. Furthermore, FA decreased MOR latency towards aliens but not towards nestmates. As response latency can be used as a proxy of response speed, we conclude that contrary to the prediction of the SAT theory, ants did not trade off speed against accuracy in the process of nestmate recognition.Summary statementExposure to an alarm pheromone increases both latency and accuracy of the response to recognition cues in ants