Displacement experiments provide evidence for path integration in Drosophila.

Like many other animals, insects are capable of returning to previously visited locations using path integration, which is a memory of travelled direction and distance. Recent studies suggest that Drosophila can also use path integration to return to a food reward. However, the existing experimental evidence for path integration in Drosophila has a potential confound: pheromones deposited at the site of reward might enable flies to find previously rewarding locations even without memory. Here, we show that pheromones can indeed cause naïve flies to accumulate where previous flies had been rewarded in a navigation task. Therefore, we designed an experiment to determine if flies can use path integration memory despite potential pheromonal cues by displacing the flies shortly after an optogenetic reward. We found that rewarded flies returned to the location predicted by a memory-based model. Several analyses are consistent with path integration as the mechanism by which flies returned to the reward. We conclude that although pheromones are often important in fly navigation and must be carefully controlled for in future experiments, Drosophila may indeed be capable of performing path integration.

High-speed locomotion in the Saharan silver ant, Cataglyphis bombycina.

The diurnal thermophilic Saharan silver ant, Cataglyphis bombycina, is the fastest of the North African Cataglyphis desert ant species. These highly mobile ants endure the extreme temperatures of their sand dune environment with outstanding behavioural, physiological and morphological adaptations. Surprisingly, C. bombycina has comparatively shorter legs than its well-studied sister species Cataglyphis fortis from salt pan habitats. This holds despite the somewhat hotter surface temperatures and the more yielding sand substrate. Here, we report that C. bombycina employs a different strategy in reaching high running speeds, outperforming the fastest known runs of the longer-legged C. fortis ants. Video analysis across a broad range of locomotor speeds revealed several differences to C. fortis Shorter leg lengths are compensated for by high stride frequencies, ranging beyond 40 Hz. This is mainly achieved by a combination of short stance phases (down to 7 ms) and fast leg swing movements (up to 1400 mm s-1). The legs of one tripod group exhibit almost perfect synchrony in the timings of their lift-offs and touch-downs, and good tripod coordination is present over the entire walking speed range (tripod coordination strength values around 0.8). This near synchrony in leg movement may facilitate locomotion across the yielding sand dune substrate.

Naturalistic path integration of Cataglyphis desert ants on an air-cushioned lightweight spherical treadmill.

Air-cushioned spheres are widely used as treadmills to study behavioural and neurophysiological questions in numerous species. We describe an improved spherical treadmill design that reliably registers the path and walking behaviour of an animal walking on top of the sphere. The simple and robust set-up consists of a very light hollowed styrofoam ball supported by an air stream in a hollow half sphere and can be used indoors and outdoors. Two optical mouse sensors provided with lenses of 4.6 mm focal length detect the motion of the sphere with a temporal resolution of more than 200 frames s-1 and a spatial resolution of less than 0.2 mm. The treadmill can be used in an open- or closed-loop configuration with respect to yaw of the animal. The tethering allows animals to freely adjust their body posture and in the closed-loop configuration to quickly rotate around their yaw axis with their own moment of inertia. In this account, we present the first evidence of naturalistic homing navigation on a spherical treadmill for two species of Cataglyphis desert ants. We were able to evaluate with good precision the walking speed and angular orientation at any time. During homing the ants showed a significant difference in walking speed between the approach and search phases; moreover, they slowed down significantly as soon as they reached zero vector state, the fictive nest position.

How to find home backwards? Navigation during rearward homing of Cataglyphis fortis desert ants.

Cataglyphis ants are renowned for their impressive navigation skills, which have been studied in numerous experiments during forward locomotion. However, the ants' navigational performance during backward homing when dragging large food loads has not been investigated until now. During backward locomotion, the odometer has to deal with unsteady motion and irregularities in inter-leg coordination. The legs' sensory feedback during backward walking is not just a simple reversal of the forward stepping movements: compared with forward homing, ants are facing towards the opposite direction during backward dragging. Hence, the compass system has to cope with a flipped celestial view (in terms of the polarization pattern and the position of the sun) and an inverted retinotopic image of the visual panorama and landmark environment. The same is true for wind and olfactory cues. In this study we analyze for the first time backward-homing ants and evaluate their navigational performance in channel and open field experiments. Backward-homing Cataglyphis fortis desert ants show remarkable similarities in the performance of homing compared with forward-walking ants. Despite the numerous challenges emerging for the navigational system during backward walking, we show that ants perform quite well in our experiments. Direction and distance gauging was comparable to that of the forward-walking control groups. Interestingly, we found that backward-homing ants often put down the food item and performed foodless search loops around the left food item. These search loops were mainly centred around the drop-off position (and not around the nest position), and increased in length the closer the ants came to their fictive nest site.

How to find home backwards? Locomotion and inter-leg coordination during rearward walking of Cataglyphis fortis desert ants.

For insects, flexibility in the performance of terrestrial locomotion is a vital part of facing the challenges of their often unpredictable environment. Arthropods such as scorpions and crustaceans can switch readily from forward to backward locomotion, but in insects this behaviour seems to be less common and, therefore, is only poorly understood. Here we present an example of spontaneous and persistent backward walking in Cataglyphis desert ants that allows us to investigate rearward locomotion within a natural context. When ants find a food item that is too large to be lifted up and to be carried in a normal forward-faced orientation, they will drag the load walking backwards to their home nest. A detailed examination of this behaviour reveals a surprising flexibility of the locomotor output. Compared with forward walks with regular tripod coordination, no main coordination pattern can be assigned to rearward walks. However, we often observed leg-pair-specific stepping patterns. The front legs frequently step with small stride lengths, while the middle and the hind legs are characterized by less numerous but larger strides. But still, these specializations show no rigidly fixed leg coupling, nor are they strictly embedded within a temporal context; therefore, they do not result in a repetitive coordination pattern. The individual legs act as separate units, most likely to better maintain stability during backward dragging.

Walking and running in the desert ant Cataglyphis fortis.

Path integration, although inherently error-prone, is a common navigation strategy in animals, particularly where environmental orientation cues are rare. The desert ant Cataglyphis fortis is a prominent example, covering large distances on foraging excursions. The stride integrator is probably the major source of path integration errors. A detailed analysis of walking behaviour in Cataglyphis is thus of importance for assessing possible sources of errors and potential compensation strategies. Zollikofer (J Exp Biol 192:95-106, 1994a) demonstrated consistent use of the tripod gait in Cataglyphis, and suggested an unexpectedly constant stride length as a possible means of reducing navigation errors. Here, we extend these studies by more detailed analyses of walking behaviour across a large range of walking speeds. Stride length increases linearly and stride amplitude of the middle legs increases slightly linearly with walking speed. An initial decrease of swing phase duration is observed at lower velocities with increasing walking speed. Then it stays constant across the behaviourally relevant range of walking speeds. Walking speed is increased by shortening of the stance phase and of the stance phase overlap. At speeds larger than 370 mm s(-1), the stride frequency levels off, the duty factor falls below 0.5, and Cataglyphis transitions to running with aerial phases.

Nest and food search behaviour in desert ants, Cataglyphis: a critical comparison.

North African desert ants, Cataglyphis, use path integration to calculate a home vector during their foraging trips, constantly informing them about their position relative to the nest. This home vector is also used to find the way back to a productive feeding site the ant has encountered and thus memorized. When the animal fails to arrive at its goal after having run off the home or food vector, a systematic search is initiated. The basic search strategies are identical for nest and food searches, consisting of a search spiral superimposed by a random walk. While nest searches have been investigated in much detail, food site searches have received comparatively little attention. Here, we quantify and compare nest and food site searches recorded under similar conditions, particularly constant nest-feeder distance, and we observe notable differences in nest and food search performances. The parameters of nest searches are relatively constant and improve little with experience, although those small improvements had not been recognized previously. Food searches, by contrast, are more flexible and cover smaller or larger areas, mainly depending on the reliability of food encounter over several visits. Intriguingly, food site searches may be significantly more focussed than nest searches, although the nest should be the most important goal in an ant's life. These results demonstrate both adaptability and high accuracy of the ants' search programme.

Establishing food site vectors in desert ants.

When returning to the site of a successful previous forage, where does one search for the goodies? Should one rely on experience from the previous homebound journey, or should one consider the outbound journey as well, or even exclusively? Desert ants are particularly well suited for pursuing this question because of their primary reliance on path integration in open and featureless desert habitats. Path integration has been studied particularly with regard to homing after lengthy foraging trips. The ants also use path integration to return to plentiful feeding sites, but what is memorised for revisiting the feeder remains controversial. Here, we demonstrate that desert ants consider, and indeed linearly average, both outbound and inbound travel for their return to a familiar feeder. This may be interpreted as a strategy to reduce navigation errors.

What counts for ants? How return behaviour and food search of Cataglyphis ants are modified by variations in food quantity and experience.

When finding more food than one is able to carry home, should one come back to the site to exploit it further? This question is crucial for central place foragers that provide for a home place with brood or nest mates. The benefit of returning has to be weighed against the chance of finding food elsewhere and the resources available. Desert ants Cataglyphis fortis are well-studied examples when it comes to navigating back and forth between their nest and a foraging area, due to their primary reliance on path integration in the open and featureless desert habitat. The ants use path integration not only for a safe return from their foraging trips but also for future returns to plentiful feeding sites. The direction from the nest that has previously yielded food items is preferred for future foraging trips, a phenomenon termed sector fidelity. What prompts the ants to return to a particular site, and how faithfully they search for that place, has not been well studied. We examine the evaluation of food sources in channel experiments by varying both the number of food items in a feeder and the number of visits to the feeder before testing search distances of foragers returning to the feeding site. Ants exhibited more focused searches for plentiful food sources than for sources with only few food items upon their first return visit. After several successful visits, the ants always searched thoroughly for the food source, independent of the amount of food offered. Thus, desert ants consider both food abundance and reliability of food encounter, with corroborative learning of reliability gradually overriding the initial preference for plentiful feeders. The density of food items appears to be used by the ants as a proxy for food abundance. On the level of our analysis, the searches performed in the experimental channels are indistinguishable from those performed in the open desert terrain. The present results not only demonstrate how otherwise well-studied desert ants assess yield and experience with reliability of food sources, but also establish a model system for future study of how itemised food sources are exploited.

Estimation of homing distance in desert ants, Cataglyphis fortis, remains unaffected by disturbance of walking behaviour.

Desert ants, Cataglyphis fortis, use a stride integrator as a distance gauge in their well-studied path integration system (while a skylight compass provides the direction gauge). To further scrutinize the mechanisms of the ant odometer, we tried to disturb the stride integrator by interfering with normal walking behaviour. First, legs that contribute to one of the two leg tripods alternately used in normal walking were selectively amputated. This prevented the normal tripod gait and should interfere with both the normal walking programme controlled by the central nervous system, and normal sensory feedback from the legs. Second, manipulation of the walking substrate in the form of regular corrugations was observed to interfere with normal walking behaviour, at least for corrugation wavelengths (12-25 mm) in the range of normal stride lengths. The animals fell and stumbled, or footfall patterns were entrained to the corrugation wavelength. The relationship between stride length and stride frequency was altered in several situations. Surprisingly, distance estimation and homing performance remained virtually unaffected even by the most severe interferences with walking behaviour. This demonstrates a remarkable robustness of walking behaviour and homing, and it suggests that stride length is determined by robust signals of leg sense organs.

Two distance memories in desert ants-Modes of interaction.

Navigation plays an essential role for many animals leading a mobile mode of life, and for central place foragers in particular. One important prerequisite for navigation is the ability to estimate distances covered during locomotion. It has been shown that Cataglyphis desert ants, well-established model organisms in insect navigation, use two odometer mechanisms, namely, stride and optic flow integration. Although both mechanisms are well established, their mode of interaction to build one odometer output remains enigmatic. We tackle this problem by selectively covering the ventral eye parts in Cataglyphis fortis foragers, the eye regions responsible for optic flow input in odometry. Exclusion of optic flow cues was implemented during different sections of outbound and inbound travel. This demonstrated that the two odometers have separate distance memories that interact in determining homing distance. Possible interpretations posit that the two odometer memories (i) take on different relative weights according to context or (ii) compete in a winner-take-all mode. Explanatory values and implications of such interpretations are discussed. We are able to provide a rough quantitative assessment of odometer cue interaction. An understanding of the interaction of different odometer mechanisms appears valuable not only for animal navigation research but may inform discussions on sensor fusion in both behavioural contexts and potential technical applications.

Optic flow odometry operates independently of stride integration in carried ants.

Cataglyphis desert ants are impressive navigators. When the foragers roam the desert, they employ path integration. For these ants, distance estimation is one key challenge. Distance information was thought to be provided by optic flow (OF)-that is, image motion experienced during travel-but this idea was abandoned when stride integration was discovered as an odometer mechanism in ants. We show that ants transported by nest mates are capable of measuring travel distance exclusively by the use of OF cues. Furthermore, we demonstrate that the information gained from the optic flowmeter cannot be transferred to the stride integrator. Our results suggest a dual information channel that allows the ants to measure distances by strides and OF cues, although both systems operate independently and in a redundant manner.

Homing distance in desert ants, Cataglyphis fortis, remains unaffected by disturbance of walking behaviour and visual input.

Desert ants gauge walking distance by means of a stride integrator and, to a minor extent, by optic flow integration. With the present experiments we attempt to interfere with both, stride integration and optic flow input in order to reveal possible interactions of the two modes of odometry and further functional details of the stride integrator. We tried to impair stride integration by amputating two of the six walking legs. Amputation of left middle and right hind legs had especially severe effects since it left only the right front leg in one of the support tripods that are alternately used in walking. We tried to impair optic flow input - which is used for distance estimation to a minor extent - by covering both ventral eye halves. These two sets of manipulations were carried out in combination to study possible compensatory effects, for instance, of optic flow input in the case of an impaired stride integrator. Unexpectedly, none of the manipulations we carried out had significant effects on homing performance. This was true with regard to homing distance estimation (as determined by the centres of the ants' nest searches) and homing certainty (as determined by the search spreads). These results corroborate the surprising robustness of odometry by stride integration, and they indicate that leg proprioceptive feedback is used for stride integration. The question of a possible interaction of optic flow input and stride integration remains open.

Re-visiting of plentiful food sources and food search strategies in desert ants.

North African desert ants, Cataglyphis fortis, are established model organisms in animal navigation research. Cataglyphis re-visit plentiful feeding sites, but their decision to return to a feeder and the organization of food searches has been little studied. Here we provide a review of recent advances regarding this topic. At least two parameters determine the ants' assessment of site quality, namely, amount of food available and reliability of food encounter on subsequent visits. The amount of food appears to be judged by the concentration of items at the food uptake site. Initially the amount of food in a feeder dominates the foragers' decision to return, whereas learning about reliability takes precedence in the course of a few visits. The location of a worthwhile site is determined by the animals' path integration system. In particular, the distance of the feeding site is memorized as the arithmetic average of the distances covered during the previous outbound and homebound journeys. Feeding sites that are small and inconspicuous cannot be approached directly with sufficient certainty, due to inevitable inaccuracies of the path integrator. Instead, desert ants steer downwind of the goal to encounter the odor plume emanating from the food and they follow this plume to the feeder. The angle steered downwind reflects the animals' maximal navigation error and is adjusted according to experience. In summary, food searches of desert ants provide an unexpected wealth of features that may advance our understanding of search, navigation, and decision strategies. There are several aspects that warrant further scrutiny.

The desert ant odometer: a stride integrator that accounts for stride length and walking speed.

Desert ants, Cataglyphis, use path integration as a major means of navigation. Path integration requires measurement of two parameters, namely, direction and distance of travel. Directional information is provided by a celestial compass, whereas distance measurement is accomplished by a stride integrator, or pedometer. Here we examine the recently demonstrated pedometer function in more detail. By manipulating leg lengths in foraging desert ants we could also change their stride lengths. Ants with elongated legs ('stilts') or shortened legs ('stumps') take larger or shorter strides, respectively, and misgauge travel distance. Travel distance is overestimated by experimental animals walking on stilts, and underestimated by animals walking on stumps - strongly indicative of stride integrator function in distance measurement. High-speed video analysis was used to examine the actual changes in stride length, stride frequency and walking speed caused by the manipulations of leg length. Unexpectedly, quantitative characteristics of walking behaviour remained almost unaffected by imposed changes in leg length, demonstrating remarkable robustness of leg coordination and walking performance. These data further allowed normalisation of homing distances displayed by manipulated animals with regard to scaling and speed effects. The predicted changes in homing distance are in quantitative agreement with the experimental data, further supporting the pedometer hypothesis.

Hair plate mechanoreceptors associated with body segments are not necessary for three-dimensional path integration in desert ants, Cataglyphis fortis.

In formicine ants, the hair fields associated with the neck and the petiole (alitrunk-petiole and petiole-gaster joints) have long been established to function in graviception. Here, we examine a possible role of these hair receptors in three-dimensional (3-D) path integration of the (formicine) desert ant, Cataglyphis fortis. Cataglyphis judge the ground distance when travelling over hills, allowing correct homing even in (unpredictably) uneven terrain. We eliminated the function of these hair sensors in graviception either by shaving the hairs or by immobilising the joints monitored by the hair plates. With that major component of their sense of graviception eliminated, one would expect the ants to disregard, or at least misgauge, the ascents and descents performed across hills during outbound journey. The ants should thus consider the (much longer) actual walking trajectory, instead of the base distance, when calculating their homing distance. Surprisingly, neither shaving nor immobilisation of the hair sensillae affected correct path integration, across both uneven terrain (3-D) and level surface. If anything, the ants underestimated homing distance, which may reflect a general, safety-oriented navigation strategy. Animals that had performed the outbound journey with their gaster fixed in a horizontal position underestimated their homing so dramatically that this latter explanation cannot hold.

The ant odometer: stepping on stilts and stumps.

Desert ants, Cataglyphis, navigate in their vast desert habitat by path integration. They continuously integrate directions steered (as determined by their celestial compass) and distances traveled, gauged by as-yet-unknown mechanisms. Here we test the hypothesis that navigating ants measure distances traveled by using some kind of step integrator, or "step counter." We manipulated the lengths of the legs and, hence, the stride lengths, in freely walking ants. Animals with elongated ("stilts") or shortened legs ("stumps") take larger or shorter strides, respectively, and concomitantly misgauge travel distance. Travel distance is overestimated by experimental animals walking on stilts and underestimated by animals walking on stumps.