Lidar as a potential tool for monitoring migratory insects.

The seasonal migrations of insects involve a substantial displacement of biomass with significant ecological and economic consequences for regions of departure and arrival. Remote sensors have played a pivotal role in revealing the magnitude and general direction of bioflows above 150 m. Nevertheless, the takeoff and descent activity of insects below this height is poorly understood. Our lidar observations elucidate the low-height dusk movements and detailed information of insects in southern Sweden from May to July, during the yearly northward migration period. Importantly, by filtering out moths from other insects based on optical information and wingbeat frequency, we have introduced a promising new method to monitor the flight activities of nocturnal moths near the ground, many of which participate in migration through the area. Lidar thus holds the potential to enhance the scientific understanding of insect migratory behavior and improve pest control strategies.

Camera-based automated monitoring of flying insects in the wild (Camfi). II. flight behaviour and long-term population monitoring of migratory Bogong moths in Alpine Australia.

Introduction: The Bogong moth Agrotis infusa is well known for its remarkable annual round-trip migration from its breeding grounds across eastern and southern Australia to its aestivation sites in the Australian Alps, to which it provides an important annual influx of nutrients. Over recent years, we have benefited from a growing understanding of the navigational abilities of the Bogong moth. Meanwhile, the population of Bogong moths has been shrinking. Recently, the ecologically and culturally important Bogong moth was listed as endangered by the IUCN Red List, and the establishment of a program for long-term monitoring of its population has been identified as critical for its conservation. Methods: Here, we present the results of two years of monitoring of the Bogong moth population in the Australian Alps using recently developed methods for automated wildlife-camera monitoring of flying insects, named Camfi. While in the Alps, some moths emerge from the caves in the evening to undertake seemingly random flights, filling the air with densities in the dozens per cubic metre. The purpose of these flights is unknown, but they may serve an important role in Bogong moth navigation. Results: We found that these evening flights occur throughout summer and are modulated by daily weather factors. We present a simple heuristic model of the arrival to and departure from aestivation sites by Bogong moths, and confirm results obtained from fox-scat surveys which found that aestivating Bogong moths occupy higher elevations as the summer progresses. Moreover, by placing cameras along two elevational transects below the summit of Mt. Kosciuszko, we found that evening flights were not random, but were systematically oriented in directions relative to the azimuth of the summit of the mountain. Finally, we present the first recorded observations of the impact of bushfire smoke on aestivating Bogong moths - a dramatic reduction in the size of a cluster of aestivating Bogong moths during the fire, and evidence of a large departure from the fire-affected area the day after the fire. Discussion: Our results highlight the challenges of monitoring Bogong moths in the wild and support the continued use of automated camera-based methods for that purpose.

Camera-based automated monitoring of flying insects (Camfi). I. Field and computational methods.

The ability to measure flying insect activity and abundance is important for ecologists, conservationists and agronomists alike. However, existing methods are laborious and produce data with low temporal resolution (e.g. trapping and direct observation), or are expensive, technically complex, and require vehicle access to field sites (e.g. radar and lidar entomology). We propose a method called "Camfi" for long-term non-invasive population monitoring and high-throughput behavioural observation of low-flying insects using images and videos obtained from wildlife cameras, which are inexpensive and simple to operate. To facilitate very large monitoring programs, we have developed and implemented a tool for automatic detection and annotation of flying insect targets in still images or video clips based on the popular Mask R-CNN framework. This tool can be trained to detect and annotate insects in a few hours, taking advantage of transfer learning. Our method will prove invaluable for ongoing efforts to understand the behaviour and ecology of declining insect populations and could also be applied to agronomy. The method is particularly suited to studies of low-flying insects in remote areas, and is suitable for very large-scale monitoring programs, or programs with relatively low budgets.

Adaptive migratory orientation of an invasive pest on a new continent.

Many species of insects undertake long-range, seasonally reversed migrations, displaying sophisticated orientation behaviors to optimize their migratory trajectories. However, when invasive insects arrive in new biogeographical regions, it is unclear if migrants retain (or how quickly they regain) ancestral migratory traits, such as seasonally preferred flight headings. Here we present behavioral evidence that an invasive migratory pest, the fall armyworm moth (Spodoptera frugiperda), a native of the Americas, exhibited locally adaptive migratory orientation less than three years after arriving on a new continent. Specimens collected from China showed flight orientations directed north-northwest in spring and southwest in autumn, and this would promote seasonal forward and return migrations in East Asia. We also show that the driver of the seasonal switch in orientation direction is photoperiod. Our results thus provide a clear example of an invasive insect that has rapidly exhibited adaptive migratory behaviors, either inherited or newly evolved, in a completely alien environment.

Stimulus-dependent orientation strategies in monarch butterflies.

Insects are well known for their ability to keep track of their heading direction based on a combination of skylight cues and visual landmarks. This allows them to navigate back to their nest, disperse throughout unfamiliar environments, as well as migrate over large distances between their breeding and non-breeding habitats. The monarch butterfly (Danaus plexippus), for instance, is known for its annual southward migration from North America to certain trees in Central Mexico. To maintain a constant flight route, these butterflies use a time-compensated sun compass, which is processed in a region in the brain, termed the central complex. However, to successfully complete their journey, the butterflies' brain must generate a multitude of orientation strategies, allowing them to dynamically switch from sun-compass orientation to a tactic behavior toward a certain target. To study whether monarch butterflies exhibit different orientation modes and if they can switch between them, we observed the orientation behavior of tethered flying butterflies in a flight simulator while presenting different visual cues to them. We found that the butterflies' behavior depended on the presented visual stimulus. Thus, while a dark stripe was used for flight stabilization, a bright stripe was fixated by the butterflies in their frontal visual field. If we replaced a bright stripe with a simulated sun stimulus, the butterflies switched their behavior and exhibited compass orientation. Taken together, our data show that monarch butterflies rely on and switch between different orientation modes, allowing the animal to adjust orientation to its actual behavioral demands.

Flight-induced compass representation in the monarch butterfly heading network.

For navigation, animals use a robust internal compass. Compass navigation is crucial for long-distance migrating animals like monarch butterflies, which use the sun to navigate over 4,000 km to their overwintering sites every fall. Sun-compass neurons of the central complex have only been recorded in immobile butterflies, and experimental evidence for encoding the animal's heading in these neurons is still missing. Although the activity of central-complex neurons exhibits a locomotor-dependent modulation in many insects, the function of such modulations remains unexplored. Here, we developed tetrode recordings from tethered flying monarch butterflies to reveal how flight modulates heading representation. We found that, during flight, heading-direction neurons change their tuning, transforming the central-complex network to function as a global compass. This compass is characterized by the dominance of processing steering feedback and allows for robust heading representation even under unreliable visual scenarios, an ideal strategy for maintaining a migratory heading over enormous distances.

A Guide for Using Flight Simulators to Study the Sensory Basis of Long-Distance Migration in Insects.

Studying the routes flown by long-distance migratory insects comes with the obvious challenge that the animal's body size and weight is comparably low. This makes it difficult to attach relatively heavy transmitters to these insects in order to monitor their migratory routes (as has been done for instance in several species of migratory birds. However, the rather delicate anatomy of insects can be advantageous for testing their capacity to orient with respect to putative compass cues during indoor experiments under controlled conditions. Almost 20 years ago, Barrie Frost and Henrik Mouritsen developed a flight simulator which enabled them to monitor the heading directions of tethered migratory Monarch butterflies, both indoors and outdoors. The design described in the original paper has been used in many follow-up studies to describe the orientation capacities of mainly diurnal lepidopteran species. Here we present a modification of this flight simulator design that enables studies of nocturnal long-distance migration in moths while allowing controlled magnetic, visual and mechanosensory stimulation. This modified flight simulator has so far been successfully used to study the sensory basis of migration in two European and one Australian migratory noctuid species.

Australian Bogong moths Agrotis infusa (Lepidoptera: Noctuidae), 1951-2020: decline and crash.

The Bogong moth Agrotis infusa is well known for its remarkable long-distance migration - a return journey from the plains of southeast Australia to the Australian Alps - as well as for its cultural significance for Indigenous Australians. Each spring, as many as four billion moths are estimated to arrive in the Australian Alps to aestivate in cool mountain caves and in boulder fields, bringing with them a massive annual influx of energy and nutrients critical for the health of the alpine ecosystem. However, a massive decline in moths present at their aestivation sites has occurred over the past 3 years, with only a few individuals present where hundreds of thousands could earlier be found. In order to understand the possible sources of decline, we analysed historical records of Bogong moth numbers at aestivation sites in the Australian Alps, including observations on Mt. Gingera (NSW) in the early 1950s, observations from 1980 onwards in the Snowy Mountains (NSW) and an almost-unbroken series of observations each summer over the past 53 years in three caves at different elevations on Mt. Buffalo (Victoria). This analysis shows that moth numbers were probably steady from 1951 until about 1980, fluctuated and slowly fell from then until 2016 and dramatically crashed in 2017. In the Murray-Darling Basin, the main winter breeding ground of Bogong moths, changes in farming practices, such as increasing land clearing for crops (which has removed around a quarter of a billion moths annually from the mountains compared to pre-European levels), has probably driven some of the decline in Bogong moth numbers observed from 1980 to 2016. The impact of insecticide remains unclear and is in urgent need of further study. Even though we found little evidence that increasing global temperatures per se are responsible for the Bogong moth decline, the Australian climate has nonetheless become drier and warmer over past decades, possibly hampering the survival of immature stages in the breeding areas and confining adult aestivation to gradually higher elevations. The crash in moth numbers from 2017 is most likely due to the recent severe drought in the moth's breeding grounds.

Spatial orientation based on multiple visual cues in non-migratory monarch butterflies.

Monarch butterflies (Danaus plexippus) are prominent for their annual long-distance migration from North America to their overwintering area in Central Mexico. To find their way on this long journey, they use a sun compass as their main orientation reference but will also adjust their migratory direction with respect to mountain ranges. This indicates that the migratory butterflies also attend to the panorama to guide their travels. Although the compass has been studied in detail in migrating butterflies, little is known about the orientation abilities of non-migrating butterflies. Here, we investigated whether non-migrating butterflies - which stay in a more restricted area to feed and breed - also use a similar compass system to guide their flights. Performing behavioral experiments on tethered flying butterflies in an indoor LED flight simulator, we found that the monarchs fly along straight tracks with respect to a simulated sun. When a panoramic skyline was presented as the only orientation cue, the butterflies maintained their flight direction only during short sequences, suggesting that they potentially use it for flight stabilization. We further found that when we presented the two cues together, the butterflies incorporate both cues in their compass. Taken together, we show here that non-migrating monarch butterflies can combine multiple visual cues for robust orientation, an ability that may also aid them during their migration.

Evidence for a southward autumn migration of nocturnal noctuid moths in central Europe.

Insect migrations are spectacular natural events and resemble a remarkable relocation of biomass between two locations in space. Unlike the well-known migrations of daytime flying butterflies, such as the painted lady (Vanessa cardui) or the monarch butterfly (Danaus plexippus), much less widely known are the migrations of nocturnal moths. These migrations - typically involving billions of moths from different taxa - have recently attracted considerable scientific attention. Nocturnal moth migrations have traditionally been investigated by light trapping and by observations in the wild, but in recent times a considerable improvement in our understanding of this phenomenon has come from studying insect orientation behaviour, using vertical-looking radar. In order to establish a new model organism to study compass mechanisms in migratory moths, we tethered each of two species of central European Noctuid moths in a flight simulator to study their flight bearings: the red underwing (Catocala nupta) and the large yellow underwing (Noctua pronuba). Both species had significantly oriented flight bearings under an unobscured view of the clear night sky and in the Earth's natural magnetic field. Red underwings oriented south-southeast, while large yellow underwings oriented southwest, both suggesting a southerly autumn migration towards the Mediterranean. Interestingly, large yellow underwings became disoriented on humid (foggy) nights while red underwings remained oriented. We found no evidence in either species for a time-independent sky compass mechanism as previously suggested for the large yellow underwing.

The Earth's Magnetic Field and Visual Landmarks Steer Migratory Flight Behavior in the Nocturnal Australian Bogong Moth.

Like many birds [1], numerous species of nocturnal moths undertake spectacular long-distance migrations at night [2]. Each spring, billions of Bogong moths (Agrotis infusa) escape hot conditions in different regions of southeast Australia by making a highly directed migration of over 1,000 km to a limited number of cool caves in the Australian Alps, historically used for aestivating over the summer [3, 4]. How moths determine the direction of inherited migratory trajectories at night and locate their destination (i.e., navigate) is currently unknown [5-7]. Here we show that Bogong moths can sense the Earth's magnetic field and use it in conjunction with visual landmarks to steer migratory flight behavior. By tethering migrating moths in an outdoor flight simulator [8], we found that their flight direction turned predictably when dominant visual landmarks and a natural Earth-strength magnetic field were turned together, but that the moths became disoriented within a few minutes when these cues were set in conflict. We thus conclude that Bogong moths, like nocturnally migrating birds [9], can use a magnetic sense. Our results represent the first reliable demonstration of the use of the Earth's magnetic field to steer flight behavior in a nocturnal migratory insect.

Corrigendum: The Australian Bogong Moth Agrotis infusa: A Long-Distance Nocturnal Navigator.

[This corrects the article on p. 77 in vol. 10, PMID: 27147998.].

Migratory blackcaps can use their magnetic compass at 5 degrees inclination, but are completely random at 0 degrees inclination.

It is known that night-migratory songbirds use a magnetic compass measuring the magnetic inclination angle, i.e. the angle between the Earth's surface and the magnetic field lines, but how do such birds orient at the magnetic equator? A previous study reported that birds are completely randomly oriented in a horizontal north-south magnetic field with 0° inclination angle. This seems counter-intuitive, because birds using an inclination compass should be able to separate the north-south axis from the east-west axis, so that bimodal orientation might be expected in a horizontal field. Furthermore, little is known about how shallow inclination angles migratory birds can still use for orientation. In this study, we tested the magnetic compass orientation of night-migratory Eurasian blackcaps (Sylvia atricapilla) in magnetic fields with 5° and 0° inclination. At 5° inclination, the birds oriented as well as they did in the normal 67° inclined field in Oldenburg. In contrast, they were completely randomly oriented in the horizontal field, showing no sign of bimodality. Our results indicate that the inclination limit for the magnetic compass of the blackcap is below 5° and that these birds indeed seem completely unable to use their magnetic compass for orientation in a horizontal magnetic field.

The Australian Bogong Moth Agrotis infusa: A Long-Distance Nocturnal Navigator.

The nocturnal Bogong moth (Agrotis infusa) is an iconic and well-known Australian insect that is also a remarkable nocturnal navigator. Like the Monarch butterflies of North America, Bogong moths make a yearly migration over enormous distances, from southern Queensland, western and northwestern New South Wales (NSW) and western Victoria, to the alpine regions of NSW and Victoria. After emerging from their pupae in early spring, adult Bogong moths embark on a long nocturnal journey towards the Australian Alps, a journey that can take many days or even weeks and cover over 1000 km. Once in the Alps (from the end of September), Bogong moths seek out the shelter of selected and isolated high ridge-top caves and rock crevices (typically at elevations above 1800 m). In hundreds of thousands, moths line the interior walls of these cool alpine caves where they "hibernate" over the summer months (referred to as "estivation"). Towards the end of the summer (February and March), the same individuals that arrived months earlier leave the caves and begin their long return trip to their breeding grounds. Once there, moths mate, lay eggs and die. The moths that hatch in the following spring then repeat the migratory cycle afresh. Despite having had no previous experience of the migratory route, these moths find their way to the Alps and locate their estivation caves that are dotted along the high alpine ridges of southeastern Australia. How naïve moths manage this remarkable migratory feat still remains a mystery, although there are many potential sensory cues along the migratory route that moths might rely on during their journey, including visual, olfactory, mechanical and magnetic cues. Here we review our current knowledge of the Bogong moth, including its natural history, its ecology, its cultural importance to the Australian Aborigines and what we understand about the sensory basis of its long-distance nocturnal migration. From this analysis it becomes clear that the Bogong moth represents a new and very promising model organism for understanding the sensory basis of nocturnal migration in insects.

Weak Broadband Electromagnetic Fields are More Disruptive to Magnetic Compass Orientation in a Night-Migratory Songbird (Erithacus rubecula) than Strong Narrow-Band Fields.

Magnetic compass orientation in night-migratory songbirds is embedded in the visual system and seems to be based on a light-dependent radical pair mechanism. Recent findings suggest that both broadband electromagnetic fields ranging from ~2 kHz to ~9 MHz and narrow-band fields at the so-called Larmor frequency for a free electron in the Earth's magnetic field can disrupt this mechanism. However, due to local magnetic fields generated by nuclear spins, effects specific to the Larmor frequency are difficult to understand considering that the primary sensory molecule should be organic and probably a protein. We therefore constructed a purpose-built laboratory and tested the orientation capabilities of European robins in an electromagnetically silent environment, under the specific influence of four different oscillating narrow-band electromagnetic fields, at the Larmor frequency, double the Larmor frequency, 1.315 MHz or 50 Hz, and in the presence of broadband electromagnetic noise covering the range from ~2 kHz to ~9 MHz. Our results indicated that the magnetic compass orientation of European robins could not be disrupted by any of the relatively strong narrow-band electromagnetic fields employed here, but that the weak broadband field very efficiently disrupted their orientation.

Migratory blackcaps tested in Emlen funnels can orient at 85 degrees but not at 88 degrees magnetic inclination.

Migratory birds are known to use the Earth's magnetic field as an orientation cue on their tremendous journeys between their breeding and overwintering grounds. The magnetic compass of migratory birds relies on the magnetic field's inclination, i.e. the angle between the magnetic field lines and the Earth's surface. As a consequence, vertical or horizontal field lines corresponding to 0 or 90 deg inclination should offer no utilizable information on where to find North or South. So far, very little is known about how small the deviations from horizontal or vertical inclination are that migratory birds can detect and use as a reference for their magnetic compass. Here, we asked: what is the steepest inclination angle at which a migratory bird, the Eurasian blackcap (Sylvia atricapilla), can still perform magnetic compass orientation in Emlen funnels? Our results show that blackcaps are able to orient in an Earth's strength magnetic field with inclination angles of 67 and 85 deg, but fail to orient in a field with 88 deg inclination. This suggests that the steepest inclination angle enabling magnetic compass orientation in migratory blackcaps tested in Emlen funnels lies between 85 and 88 deg.

3D Standard Brain of the Red Flour Beetle Tribolium Castaneum: A Tool to Study Metamorphic Development and Adult Plasticity.

The red flour beetle Tribolium castaneum is emerging as a further standard insect model beside Drosophila. Its genome is fully sequenced and it is susceptible for genetic manipulations including RNA-interference. We use this beetle to study adult brain development and plasticity primarily with respect to the olfactory system. In the current study, we provide 3D standard brain atlases of freshly eclosed adult female and male beetles (A0). The atlases include eight paired and three unpaired neuropils including antennal lobes (ALs), optic lobe neuropils, mushroom body calyces and pedunculi, and central complex. For each of the two standard brains, we averaged brain areas of 20 individual brains. Additionally, we characterized eight selected olfactory glomeruli from 10 A0 female and male beetles respectively, which we could unequivocally recognize from individual to individual owing to their size and typical position in the ALs. In summary, comparison of the averaged neuropil volumes revealed no sexual dimorphism in any of the reconstructed neuropils in A0 Tribolium brains. Both, the female and male 3D standard brain are also used for interspecies comparisons, and, importantly, will serve as future volumetric references after genetical manipulation especially regarding metamorphic development and adult plasticity.

Visual but not trigeminal mediation of magnetic compass information in a migratory bird.

Magnetic compass information has a key role in bird orientation, but the physiological mechanisms enabling birds to sense the Earth's magnetic field remain one of the unresolved mysteries in biology. Two biophysical mechanisms have become established as the most promising magnetodetection candidates. The iron-mineral-based hypothesis suggests that magnetic information is detected by magnetoreceptors in the upper beak and transmitted through the ophthalmic branch of the trigeminal nerve to the brain. The light-dependent hypothesis suggests that magnetic field direction is sensed by radical pair-forming photopigments in the eyes and that this visual signal is processed in cluster N, a specialized, night-time active, light-processing forebrain region. Here we report that European robins with bilateral lesions of cluster N are unable to show oriented magnetic-compass-guided behaviour but are able to perform sun compass and star compass orientation behaviour. In contrast, bilateral section of the ophthalmic branch of the trigeminal nerve in European robins did not influence the birds' ability to use their magnetic compass for orientation. These data show that cluster N is required for magnetic compass orientation in this species and indicate that it may be specifically involved in processing of magnetic compass information. Furthermore, the data strongly suggest that a vision-mediated mechanism underlies the magnetic compass in this migratory songbird, and that the putative iron-mineral-based receptors in the upper beak connected to the brain by the trigeminal nerve are neither necessary nor sufficient for magnetic compass orientation in European robins.