Asymmetry between the dorsal and ventral digging valves of the female locust: function and mechanics.

BACKGROUND: The female locust is equipped with unique digging tools, namely two pairs of valves-a dorsal and a ventral-utilized for excavating an underground hole in which she lays her eggs. This apparatus ensures that the eggs are protected from potential predators and provides optimal conditions for successful hatching. The dorsal and the ventral valves are assigned distinct roles in the digging process. Specifically, the ventral valves primarily function as anchors during propagation, while the dorsal valves displace soil and shape the underground tunnel. RESULTS: In this study, we investigated the noticeable asymmetry and distinct shapes of the valves, using a geometrical model and a finite element method. Our analysis revealed that although the two pairs of valves share morphological similarities, they exhibit different 3D characteristics in terms of absolute size and structure. We introduced a structural characteristic, the skew of the valve cross-section, to quantify the differences between the two pairs of valves. Our findings indicate that these structural variations do not significantly contribute to the valves' load-bearing capabilities under external forces. CONCLUSIONS: The evolutionary development of the form of the female locust digging valves is more aligned with fitting their respective functions rather than solely responding to biomechanical support needs. By understanding the intricate features of these locust valves, and using our geometrical model, valuable insights can be obtained for creating more efficient and specialized tools for various digging applications.

Target for lipid-to-carbohydrate intake minimizes cost of growth.

Many theoretical treatments of foraging use energy as currency, with carbohydrates and lipids considered interchangeable as energy sources. However, herbivores must often synthesize lipids from carbohydrates since they are in short supply in plants, theoretically increasing the cost of growth. We tested whether a generalist insect herbivore (Locusta migratoria) can improve its growth efficiency by consuming lipids, and whether these locusts have a preferred caloric intake ratio of carbohydrate to lipid (C : L). Locusts fed pairs of isocaloric, isoprotein diets differing in C and L consistently selected a 2C : 1L target. Locusts reared on isocaloric, isoprotein 3C : 0L diets attained similar final body masses and lipid contents to locusts fed the 2C : 1L diet, but they ate more and had a ~12% higher metabolic rate, indicating an energetic cost for lipogenesis. These results demonstrate that some animals can selectively regulate carbohydrate-to-lipid intake and that consumption of dietary lipids can improve growth efficiency.

Theoretical principles explain the structure of the insect head direction circuit.

To navigate their environment, insects need to keep track of their orientation. Previous work has shown that insects encode their head direction as a sinusoidal activity pattern around a ring of neurons arranged in an eight-column structure. However, it is unclear whether this sinusoidal encoding of head direction is just an evolutionary coincidence or if it offers a particular functional advantage. To address this question, we establish the basic mathematical requirements for direction encoding and show that it can be performed by many circuits, all with different activity patterns. Among these activity patterns, we prove that the sinusoidal one is the most noise-resilient, but only when coupled with a sinusoidal connectivity pattern between the encoding neurons. We compare this predicted optimal connectivity pattern with anatomical data from the head direction circuits of the locust and the fruit fly, finding that our theory agrees with experimental evidence. Furthermore, we demonstrate that our predicted circuit can emerge using Hebbian plasticity, implying that the neural connectivity does not need to be explicitly encoded in the genetic program of the insect but rather can emerge during development. Finally, we illustrate that in our theory, the consistent presence of the eight-column organisation of head direction circuits across multiple insect species is not a chance artefact but instead can be explained by basic evolutionary principles.

Insects, including fruit flies and locusts, move throughout their environment to find food, interact with each other or escape danger. To navigate their surroundings, insects need to be able to keep track of their orientation. This tracking is achieved through visual cues and integrating information about their movements whilst flying so they know which direction their head is facing. The set of neurons responsible for relaying information about the direction of the head (also known as heading) are connected together in a ring made up of eight columns of cells. Previous studies showed that the level of activity across this ring of neurons resembles a sinusoid shape: a smooth curve with one peak which encodes the animal's heading. Neurons downstream from this eight-column ring, which relay velocity information, also display this sinusoidal pattern of activation. Aceituno, Dall'Osto and Pisokas wanted to understand whether this sinusoidal pattern was an evolutionary coincidence, or whether it offers a particular advantage to insects. To answer this question, they established the mathematical criteria required for neurons in the eight-column ring to encode information about the heading of the animal. This revealed that these conditions can be satisfied by many different patterns of activation, not just the sinusoidal shape. However, Aceituno, Dall'Osto and Pisokas show that the sinusoidal shape is the most resilient to variations in neuronal activity which may impact the encoded information. Further experiments revealed that this resilience only occurred if neurons in the circuit were connected together in a certain pattern. Aceituno, Dall'Osto and Pisokas then compared this circuit with experimental data from locusts and fruit flies and found that both insects exhibit the predicted connection pattern. They also discovered that animals do not have to be born with this neuronal connection pattern, but can develop it during their lifetime. These findings provide fresh insights into how insects relay information about the direction of their head as they fly. They suggest that the structure of the neuronal circuit responsible for encoding head direction was not formed by chance but instead arose due to the evolutionary benefits it provided.

Risk of migration: not all gregarious locust nymphs reach appropriate refuges.

Sedentary animals choose appropriate refuges against predators, while migratory ones may not necessarily do so. In ectotherms, refuge selection is critical during low temperatures, because they cannot actively evade predators. To understand how migratory ectotherms alter their defensive behaviors depending on refuge quality in cold temperatures, we evaluated migratory gregarious desert locust nymphs (Schistocerca gregaria) in the Sahara Desert, where daily thermal constraints occur. We recorded how roosting plant type (bush/shrub) and its height influenced two alternative defense behaviors (dropping/stationary) during cold mornings, in response to an approaching simulated ground predator. Most locusts in bushes dropped within the bush and hid irrespective of their height, whereas those roosting > 2 m height in shrubs remained stationary. These defenses are effective and match with refuge plant types because dynamic locomotion is not required. When nymphs roosted on shrubs < 1.5-m height, which was an unsafe position, nearly half showed both defensive behaviors, indicating that escaping decisions become ambiguous when the refuges are inappropriate. These results suggest that locusts display flexible defensive behaviors when finding appropriate refuges and selecting refuge before daily thermal limitations occur could be critical for migratory ectotherms, which is a risk associated with migration.

Parental experiences orchestrate locust egg hatching synchrony by regulating nuclear export of precursor miRNA.

Parental experiences can affect the phenotypic plasticity of offspring. In locusts, the population density that adults experience regulates the number and hatching synchrony of their eggs, contributing to locust outbreaks. However, the pathway of signal transmission from parents to offspring remains unclear. Here, we find that transcription factor Forkhead box protein N1 (FOXN1) responds to high population density and activates the polypyrimidine tract-binding protein 1 (Ptbp1) in locusts. FOXN1-PTBP1 serves as an upstream regulator of miR-276, a miRNA to control egg-hatching synchrony. PTBP1 boosts the nucleo-cytoplasmic transport of pre-miR-276 in a "CU motif"-dependent manner, by collaborating with the primary exportin protein exportin 5 (XPO5). Enhanced nuclear export of pre-miR-276 elevates miR-276 expression in terminal oocytes, where FOXN1 activates Ptbp1 and leads to egg-hatching synchrony in response to high population density. Additionally, PTBP1-prompted nuclear export of pre-miR-276 is conserved in insects, implying a ubiquitous mechanism to mediate transgenerational effects.

Retention Rates of Genetic Therapies Based on AAV Serotypes 2 and 8 Using Different Drug-Delivery Materials.

The purpose of this study was to compare the retention rate of Adeno-associated viral vector (AAV) gene therapy agents within different subretinal injection systems. The retention of AAV serotype 2-based voretigene neparvovec (VN) and a clinical-grade AAV serotype 8 vector within four different subretinal cannulas from two different manufacturers was quantified. A standardized qPCR using the universal inverted terminal repeats as a target sequence was developed. The instruments compared were the PolyTip® cannula 25 g/38 g by MedOne Surgical, Inc., Sarasota, FL, USA, and three different subretinal injection needles by DORC, Zuidland, The Netherlands (1270.EXT Extendible 41G subretinal injection needle (23G), DORC 1270.06 23G Dual bore injection cannula, DORC 27G Subretinal injection cannula). The retention rate of VN and within the DORC products (10-28%) was comparable to the retention rate (32%) found for the PolyTip® cannula that is mentioned in the FDA-approved prescribing information for VN. For the AAV8 vector, the PolyTip® cannula showed a retention rate of 14%, and a similar retention rate of 3-16% was found for the DORC products (test-retest variability: mean 4.5%, range 2.5-20.2%). As all the instruments tested showed comparable retention rates, they seem to be equally compatible with AAV2- and AAV8-based gene therapy agents.

Insights into the molecular mechanism of yellow cuticle coloration by a chitin-binding carotenoprotein in gregarious locusts.

Carotenoids are hydrophobic pigments binding to diverse carotenoproteins, many of which remain unexplored. Focusing on yellow gregarious locusts accumulating cuticular carotenoids, here we use engineered Escherichia coli cells to reconstitute a functional water-soluble ß-carotene-binding protein, BBP. HPLC and Raman spectroscopy confirmed that recombinant BBP avidly binds ß-carotene, inducing the unusual vibronic structure of its absorbance spectrum, just like native BBP extracted from the locust cuticles. Bound to recombinant BBP, ß-carotene exhibits pronounced circular dichroism and allows BBP to withstand heating (T0.5 = 68 °C), detergents and pH variations. Using bacteria producing distinct xanthophylls we demonstrate that, while ß-carotene is the preferred carotenoid, BBP can also extract from membranes ketocarotenoids and, very poorly, hydroxycarotenoids. We show that BBP-carotenoid complex reversibly binds to chitin, but not to chitosan, implying the role for chitin acetyl groups in cuticular BBP deposition. Reconstructing such locust coloration mechanism in vitro paves the way for structural studies and BBP applications.

Molecular strategies of the pygmy grasshopper Eucriotettix oculatus adapting to long-term heavy metal pollution.

To study the heavy metal accumulation and its impact on insect exterior and chromosome morphology, and reveal the molecular mechanism of insects adapting to long-term heavy metal compound pollution habitats, this study, in the Diaojiang river basin, which has been polluted by heavy metals(HMs) for nearly a thousand years, two Eucriotettix oculatus populations was collected from mining and non-mining areas. It was found that the contents of 7 heavy metals (As, Cd, Pb, Zn, Cu, Sn, Sb) in E. oculatus of the mining area were higher than that in the non-mining 1-11 times. The analysis of morphology shows that the external morphology, the hind wing type and the chromosomal morphology of E. oculatus are significant differences between the two populations. Based on the heavy metal accumulation，morphological change, and stable population density, it is inferred that the mining area population has been affected by heavy metals and has adapted to the environment of heavy metals pollution. Then, by analyzing the transcriptome of the two populations, it was found that the digestion, immunity, excretion, endocrine, nerve, circulation, reproductive and other systems and lysosomes, endoplasmic reticulum and other cell structure-related gene expression were suppressed. This shows that the functions of the above-mentioned related systems of E. oculatus are inhibited by heavy metal stress. However, it has also been found that through the significant up-regulation of genes related to the above system, such as ATP2B, pepsin A, ubiquitin, AQP1, ACOX, ATPeV0A, SEC61A, CANX, ALDH7A1, DLD, aceE, Hsp40, and catalase, etc., and the down-regulation of MAPK signalling pathway genes, can enhanced nutrient absorption, improve energy metabolism, repair damaged cells and degrade abnormal proteins, maintain the stability of cells and systems, and resist heavy metal damage so that E. oculatus can adapt to the environment of heavy metal pollution for a long time.

Chromosome-level genome assembly of the pygmy grasshopper Eucriotettix oculatus (Orthoptera: Tetrigoidea).

The pygmy grasshoppers, which belong to the superfamily Tetrigoidea, exhibit remarkable environmental adaptability. However, no study has yet reported a reference genome for this group. In this study, we assembled a high-quality chromosome-scale genome of Eucriotettix oculatus, which survive in the environment heavily polluted by heavy metals, achieved through Illumina and PacBio sequencing, alongside chromosome conformation capture techniques. The resulting genome spans 985.45 Mb across seven chromosomes (range: 71.55 to 266.65 Mb) and features an N50 length of 123.82 Mb. Chr5 is considered to be the single sex chromosome (X). This genome is composed of 46.42% repetitive elements and contains 14,906 predicted protein-coding genes, 91.63% of which are functionally annotated. Decoding the E. oculatus genome not only promotes future studies on environmental adaptation for the pygmy grasshopper, but also provides valuable resources for in-depth investigation on phylogeny, evolution, and behavior of Orthoptera.

Myanmar endemic Burmorthacris subaptera Kevan, Singh et Akbar, 1964 (Pyrgomorphidae: Orthacridini) rediscovered after 85 years.

A curious micropteous gaudy grasshopper (family Pyrgomorphidae), Burmorthacris subaptera was described by Kevan, Singh and Akbar in 1964 as a sole member of its genus based on a female and a male collected in Yenangyaung (upper Myanmar) on 27th and 28th August 1937 and which were deposited at the Academy of Natural Sciences of Philadelphia. The species has never been reported since then. The genus Burmorthacris is the northernmost of the Orthacris genus group genera, which includes mostly genera from Sri Lanka and Malaysia. Recently we rediscovered this B. subaptera in its type locality (Yenangyaung township in the Magway Region) in Myanmar, 85 years after the holotype and the paratype were collected in the same place, and furthermore herewith we present one more locality in another region where the species has been found (Nyaung-U township in the Mandalay Region). Due to the lack of basically any information on this species distribution and habitat, including also photographs of its natural coloration in this habitat, the present study provides the first-time photographs of B. subaptera in its natural habitat from both localities, as well as some insights into its morphology, especially coloration, habitat, and behavior.

Studies on Eumastacoid grasshoppers: Paralethus montanus n. sp. (Orthoptera: Episactidae) from Chiapas, Mexico.

Paralethus montanus n. sp. is described from Chiapas, Mexico. This new species constitutes the first record of the genus for Mexico, previously known only for El Salvador and Guatemala, and it is the first episactid species collected at higher elevations (above 3000 m). A standardization for the abbreviations of the genital structures of episactids is proposed, which could apply to other groups of eumastacoids. A key for the identification of Paralethus species is also provided. Finally, the taxonomy of the genus and the spatial and altitudinal distribution of Central American taxa are discussed.

Limited genetic variability and spatial population structure in grasshoppers between natural and metal-contaminated areas in Egypt.

Pollutants in an environment can have long-term implications for the species living there, resulting in local adaptations with implications for their genetic structure. Heavy metal pollutants infiltrate soils and groundwater, bioaccumulate in food webs, and negatively impact biota. In this study, we investigated the degree to which the genetic structure and variability of the slender green-winged grasshopper (Aiolopus thalassinus (Fabricius) (Orthoptera: Acrididae)) were impacted by heavy metal pollution and distance. We used the random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) method to examine the genetic variability of populations in 3 heavy metal-polluted and 3 unpolluted locations across varying geographical distances in Egypt. The heavy metal concentrations of cadmium, copper, lead, and zinc were measured from the grasshopper tissue and soils. Sixty-nine unique and polymorphic bands were produced by 4 primers. Cluster and principal component analyses separated the populations inside and outside Cairo into 2 main branches, which were further divided into smaller branches corresponding to their geographical regions. We found no differences in the Shannon genetic diversity index between populations or with increasing heavy metal concentrations in either the soil or the grasshopper tissue. Our results showed a greater genetic variation among populations than between populations within the same location, indicating populations within locations were less differentiated than those between locations. The moderate correlation between genetic similarity and spatial distance suggests geographical isolation influenced grasshopper population differentiation. Based on the RAPD analysis, environmental pollutants and geographical distances impact the A. thalassinus population structure, potentially restricting gene flow between sites even at small spatial scales.

Kinetic characterization of a flavin-dependent monooxygenase from the insect food crop pest, Zonocerus variegatus.

Zonocerus variegatus, or the painted grasshopper, is a food crop pest endemic in Western and Central Africa. Agricultural industries in these regions rely heavily on natural defense mechanisms to control the grasshopper population such as plant-secreted alkaloid compounds. In recent years, the Z. variegatus population has continued to rise due to acquired resistance to alkaloids. Here we focus on the kinetic characterization of a flavin-dependent monooxygenase, ZvFMO, that catalyzes the nitrogen oxidation of many of these alkaloid compounds and confers resistance to the insect. Expression and purification of ZvFMO through a traditional E. coli expression system was successful and provided a unique opportunity to characterize the catalytic properties of an FMO from insects. ZvFMO was found to catalyze oxidation reactions of tertiary nitrogen atoms and the sulfur of cysteamine. Using stopped-flow spectroscopy, we have determined the kinetic mechanism of ZvFMO. We assessed F383 for its involvement in substrate binding, which was previously proposed, and determined that this residue does not play a major role in binding substrates. Through molecular docking, we identified N304 and demonstrated that this residue plays a role in substrate binding. The role of K215 was studied and was shown that it plays a critical role in NAD(P)H binding and cofactor selectivity.

A neural m6A pathway regulates behavioral aggregation in migratory locusts.

RNA N6-methyladenosine (m6A), as the most abundant modification of messenger RNA, can modulate insect behaviors, but its specific roles in aggregation behaviors remain unexplored. Here, we conducted a comprehensive molecular and physiological characterization of the individual components of the methyltransferase and demethylase in the migratory locust Locusta migratoria. Our results demonstrated that METTL3, METTL14 and ALKBH5 were dominantly expressed in the brain and exhibited remarkable responses to crowding or isolation. The individual knockdown of methyltransferases (i.e., METTL3 and METTL14) promoted locust movement and conspecific attraction, whereas ALKBH5 knockdown induced a behavioral shift toward the solitary phase. Furthermore, global transcriptome profiles revealed that m6A modification could regulate the orchestration of gene expression to fine tune the behavioral aggregation of locusts. In summary, our in vivo characterization of the m6A functions in migratory locusts clearly demonstrated the crucial roles of the m6A pathway in effectively modulating aggregation behaviors.

Study on the prediction method of grasshopper occurrence risk in Inner Mongolia based on the maximum entropy model during the growing period.

Grasshoppers represent a significant biological challenge in Inner Mongolia's grasslands, severely affecting the region's animal husbandry. Thus, dynamic monitoring of grasshopper infestation risk is crucial for sustainable livestock farming. This study employed the Maxent model, along with remote sensing data, to forecast Oedaleus decorus asiaticus occurrence during the growing season, using grasshopper suitability habitats as a base. The Maxent model's predictive accuracy was high, with an AUC of 0.966. The most influential environmental variables for grasshopper distribution were suitable habitat data (34.27%), the temperature-vegetation dryness index during the spawning period (18.81%), and various other meteorological and vegetation factors. The risk index model was applied to calculate the grasshopper distribution across different risk levels for the years 2019-2022. The data indicated that the level 1 risk area primarily spans central, eastern, and southwestern Inner Mongolia. By examining the variable weights, the primary drivers of risk level fluctuation from 2019 to 2022 were identified as accumulated precipitation and land surface temperature anomalies during the overwintering period. This study offers valuable insights for future O. decorus asiaticus monitoring in Inner Mongolia.

Recent advances in insect vision in a 3D world: looming stimuli and escape behaviour.

Detecting looming motion directly towards the insect is vital to its survival. Looming detection in two insects, flies and locusts, is described and contrasted. Pathways using looming detectors to trigger action and their topographical layout in the brain is explored in relation to facilitating behavioural selection. Similar visual stimuli, such as looming motion, are processed by nearby glomeruli in the brain. Insect-inspired looming motion detectors are combined to detect and avoid collision in different scenarios by robots, vehicles and unmanned aerial vehicle (UAV)s.

Linking shape conspicuous asymmetry with shape covariation patterns and performance in the insect head and mandibles.

Bilateral symmetry is widespread across animals, yet, among bilaterians, many cases of conspicuous asymmetries evolved. This means that bilaterally homologous structures on the left and right sides display divergent phenotypes. The evolution of such divergent phenotypes between otherwise similarly shaped structures can be thought to be favored by modularity, but this has rarely been studied in the context of left-right differences. Here, we provide an empirical example, using geometric morphometrics to assess patterns of asymmetry and covariation between landmark partitions in a grasshopper with conspicuously asymmetric mandibles. Our morphometric data confirm the presence of strictly directional conspicuous asymmetry in the mandibles and surrounding structures. Covariance patterns and tests hint at a strong integration between mandibles despite their divergent morphologies, and variational modularity with the head capsule. While mandibles have been selected to achieve a key-and-lock morphology by having interlocking shapes, the developmental modularity required to achieve this seems to be overwritten by developmental and/or functional integration, allowing the precise matching required for feeding. The consequent conflicting covariation patterns are reminiscent of the palimpsest model. Finally, the degree of directional asymmetry appears to be under selection, although we find no relationship between bite force and mandible shape or asymmetry.

Multiple forces facilitate the aquatic acrobatics of grasshopper and bioinspired robot.

Aquatic locomotion is challenging for land-dwelling creatures because of the high degree of fluidity with which the water yields to loads. We surprisingly found that the Chinese rice grasshopper Oxya chinensis, known for its terrestrial acrobatics, could swiftly launch itself off the water's surface in around 25 ms and seamlessly transition into flight. Biological observations showed that jumping grasshoppers use their front and middle legs to tilt up bodies first and then lift off by propelling the water toward the lower back with hind legs at angular speeds of up to 18°/ms, whereas the swimming grasshoppers swing their front and middle legs in nearly horizontal planes and move hind legs less violently (~8°/ms). Force measurement and model analysis indicated that the weight support could be achieved by hydrostatics which are proportionate to the mass of the grasshoppers, while the propulsions for motion are derived from the controlled limb-water interactions (i.e., the hydrodynamics). After learning the structural and behavioral strategies of the grasshoppers, a robot was created and was capable of swimming and jumping on the water surface like the insects, further demonstrating the effectiveness of decoupling the challenges of aquatic locomotion by the combined use of the static and dynamic hydro forces. This work not only uncovered the combined mechanisms responsible for facilitating aquatic acrobatics in this species but also laid a foundation for developing bioinspired robots that can locomote across multiple media.

Pausing to swarm: locust intermittent motion is instrumental for swarming-related visual processing.

Intermittent motion is prevalent in animal locomotion. Of special interest is the case of collective motion, in which social and environmental information must be processed in order to establish coordinated movement. We explored this nexus in locust, focusing on how intermittent motion interacts with swarming-related visual-based decision-making. Using a novel approach, we compared individual locust behaviour in response to continuously moving stimuli, with their response in semi-closed-loop conditions, in which the stimuli moved either in phase with the locust walking, or out of phase, i.e. only during the locust's pauses. Our findings clearly indicate the greater tendency of a locust to respond and 'join the swarming motion' when the visual stimuli were presented during its pauses. Hence, the current study strongly confirms previous indications of the dominant role of pauses in the collective motion-related decision-making of locusts. The presented insights contribute to a deeper general understanding of how intermittent motion contributes to group cohesion and coordination in animal swarms.