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 We studied the songs and morphology of the stridulatory file of Isophya modestior across its complete geographic range, in order to test our hypothesis that the male calling song of the species shows strong differentiation between the northern (Pannonian) and southern (Balkan) parts of its distribution range, reflecting its disjunct distribution. Our analyses confirm this hypothesis, separating analyzed specimens of I. modestior into two main groups - one present in the central part of the Balkan Peninsula (representing Isophya modestior sensu stricto ), with the second group occurring in the Pannonian Basin, Dinarides, Slovenia and NE Italy. The most reliable difference between the groups is the duration of the main syllable, the number of stridulatory teeth and number of pulses in the main syllable, where all values are higher in specimens from the Balkan Peninsula. Additional analyses showed that within the second group, there are differences in analyzed characters between specimens from the Pannonian Basin and specimens from the Dinaric area, the latter ones having intermediate song characteristics, closer to the group from the Balkan Peninsula. Our study shows that detailed bioacoustic analyses can help to unravel patterns of intraspecific differentiation and thus provide a useful tool for taxonomic studies. Keywords: Balkans, bioacoustics, bush-cricket, oscillogram, Pannonian Basin, stridulatory file

 The Phaneropterinae, commonly known as the bush katydids, are among the most diverse tettigoniids in the world. A new species Anormalous liu sp. nov. is described from Kashmir, India. This is the second species in the short-winged genus Anormalous . It is differentiated from the other species from China by the absence of posterior apical spurs on the fore and mid tibiae, the male subgenital plate with two long cylindrical lobes fused with each other and blunt at the apices, and the male stridulatory area longer than broad. We include a key to species in the genus Anormalou . The holotype has been deposited in the Museum of Zoology Department, Aligarh Muslim University, Aligarh Uttar Pradesh, India. Keywords: Anormalous , India, Kashmir, new species, Phaneropterinae

Mammalian hearing operates on three basic steps: 1) sound capturing, 2) impedance conversion, and 3) frequency analysis. While these canonical steps are vital for acoustic communication and survival in mammals, they are not unique to them. An equivalent mechanism has been described for katydids (Insecta), and it is unique to this group among invertebrates. The katydid inner ear resembles an uncoiled cochlea, and has a length less than 1 mm. Their inner ears contain the crista acustica , which holds tonotopically arranged sensory cells for frequency mapping via travelling waves. The crista acustica is located on a curved triangular surface formed by the dorsal wall of the ear canal. While empirical recordings show tonotopic vibrations in the katydid inner ear for frequency analysis, the biophysical mechanism leading to tonotopy remains elusive due to the small size and complexity of the hearing organ. In this study, robust numerical simulations are developed for an in silico investigation of this process. Simulations are based on the precise katydid inner ear geometry obtained by synchrotron-based micro-computed tomography, and empirically determined inner ear fluid properties for an accurate representation of the underlying mechanism. We demonstrate that the triangular structure below the hearing organ drives the tonotopy and travelling waves in the inner ear, and thus has an equivalent role to the mammalian basilar membrane. This reveals a stronger analogy between the inner ear basic mechanical networks of two organisms with ancient evolutionary differences and independent phylogenetic histories.

Curated DNA barcode reference libraries are crucial for advancing environmental DNA (eDNA) studies, monitoring biological invasions, reliable biodiversity assessments, accurate species identification, etc. However, DNA barcode databases remain highly incomplete for most invertebrate taxa. In this study, we present the most comprehensive reference library to date for the family Tettigoniidae (Orthoptera) from the Iberian Peninsula—the most species-rich orthopteran family globally, with over 8,000 valid species. We generated 402 new DNA barcodes from at least 121 tettigoniid species from the Iberian Peninsula and integrated these with 169 previously published sequences. The resulting dataset comprises 571 barcoded specimens, representing 49 genera and 123 species, including many recently described taxa. Notably, we provide DNA barcodes for at least 68 described species that previously lacked them. Our dataset covers 85% of the tettigoniid species in the Iberian Peninsula and approximately 25% of European bush-cricket species. Furthermore, our analyses show that most tettigoniid species (95%) can be reliably identified using DNA barcoding. However, mitochondrial introgression events were detected in several species of the subfamilies Bradyporinae and Tettigoniinae, highlighting the need for cautious application of this molecular identification tool.

Background DNA barcoding has been developed as a useful tool for species discrimination. Several sequence-based species delimitation methods, such as Barcode Index Number (BIN), REfined Single Linkage (RESL), Automatic Barcode Gap Discovery (ABGD), a Java program uses an explicit, determinate algorithm to define Molecular Operational Taxonomic Unit (jMOTU), Generalized Mixed Yule Coalescent (GMYC), and Bayesian implementation of the Poisson Tree Processes model (bPTP), were used. Our aim was to estimate Chinese katydid biodiversity using standard DNA barcode cytochrome c oxidase subunit I (COI-5P) sequences. Results Detection of a barcoding gap by similarity-based analyses and clustering-base analyses indicated that 131 identified morphological species (morphospecies) were assigned to 196 BINs and were divided into four categories: (i) MATCH (83/131 = 64.89%), morphospecies were a perfect match between morphospecies and BINs (including 61 concordant BINs and 22 singleton BINs); (ii) MERGE (14/131 = 10.69%), morphospecies shared its unique BIN with other species; (iii) SPLIT (33/131 = 25.19%, when 22 singleton species were excluded, it rose to 33/109 = 30.28%), morphospecies were placed in more than one BIN; (iv) MIXTURE (4/131 = 5.34%), morphospecies showed a more complex partition involving both a merge and a split. Neighbor-joining (NJ) analyses showed that nearly all BINs and most morphospecies formed monophyletic cluster with little variation. The molecular operational taxonomic units (MOTUs) were defined considering only the more inclusive clades found by at least four of seven species delimitation methods. Our results robustly supported 61 of 109 (55.96%) morphospecies represented by more than one specimen, 159 of 213 (74.65%) concordant BINs, and 3 of 8 (37.5%) discordant BINs. Conclusions Molecular species delimitation analyses generated a larger number of MOTUs compared with morphospecies. If these MOTU splits are proven to be true, Chinese katydids probably contain a seemingly large proportion of cryptic/undescribed taxa. Future amplification of additional molecular markers, particularly from the nuclear DNA, may be especially useful for specimens that were identified here as problematic taxa.

Ecotones occur naturally throughout complex landscapes. Each ecotone has particular ecological conditions resulting in species-specific responses. Across anthropogenic landscape mosaics both natural and cultural processes maintain ecotones. However, there is also a behavioural component associated with ecotones, allowing fine interpretation of ecotones in terms of animal responses, where traditional sampling has disadvantages. Singing bush crickets are often major components of soundscapes, with individuals having strong associations with certain vegetation types. Here we use ecoacoustic methods to determine how singing bush cricket species characterize the soundscape of natural vs. anthropogenic ecotones in a complex landscape. We compare the response of bush crickets to anthropogenic and natural ecotones based on acoustic species composition, acoustic activity, and total call times in response to measured vegetation characteristics. Eleven bush cricket species were identified from almost 90,000 calls from both natural and anthropogenic biotope interiors and their ecotones. Both the natural and anthropogenic ecotones were almost identical in assemblage composition, were diverse, and further supported high degrees of bush cricket singing activity. The bush cricket assemblages at these ecotones did not differ, yet they proved to be the turnover point between assemblages in the open vs. woodier sites. This complex landscape supported considerable environmental heterogeneity, reflected in the soundscape. Interestingly, we show here that both natural and anthropogenic ecotones have value for bush crickets in this semi-transformed landscape.

Wind, a major source of environmental noise, forces invertebrates that communicate with plant-borne vibrations to adjust their signaling when communicating in windy conditions. However, the strategies that animals use to reduce the impact of wind noise on communication are not well studied. We investigated the effects of wind on the production of tremulatory signals in the neotropical katydid Copiphora brevirostris. First, we recorded katydid signaling activity and natural wind variation in the field. Additionally, we exposed katydid couples during their most active signaling time period to artificial wind of different levels, and we recorded the number of tremulations produced by the males. We found that wind levels are at their lowest between 2:00 and 5:00 in the morning, which coincides with peak signaling period for male katydids. Furthermore, we found that males produce significantly fewer tremulations when exposed to wind rather than acoustic noise or silence. Wind velocity significantly affected the number of tremulations produced during the wind treatment, with fewer tremulations produced with higher wind velocities. Our results show that katydids can time their vibratory signaling both in the short-and long-term to favorable sensory conditions, either through behavioral flexibility in response to short-term fluctuations in wind or as a result of an evolutionary process in response to predictable periods of low-wind conditions.

Gampsocleis gratiosa Brunner von Wattenwyl, 1862, is a type of omnivorous chirping insect with a long history of artificial breeding. It has high economic value and is also an excellent orthopteran model organism. In this study, 12 types of culture media combined with 16S rRNA sequencing were employed to isolate 838 bacterial strains from the gut of G. gratiosa. After sequence comparison, a total of 98 species of bacteria were identified, belonging to 3 phyla, 5 classes, 11 orders, 20 families, and 45 genera. Firmicutes and Proteobacteria accounted for the majority (92.86%). At the order level, Enterobacteriaceae, Bacillales, and Lactobacillales predominated (79.59%). At the genus level, Klebsiella (11.22%) and Enterococcus (7.14%) predominated. This study also enumerated the strain morphological, physiological and biochemical properties of 98 species of bacteria, including colony morphology, Gram staining, bacterial motility test, temperature gradient growth, pH gradient growth, citrate utilization test, temperature oxidase test, contact enzyme test, methyl red test, V-P test, indole test, gelatin liquefaction test, nitrate reduction test, hydrogen sulfide test, starch hydrolysis test, cellulose decomposition test, esterase (corn oil) test and antibiotic susceptibility testing. Additionally, 16 antibiotics were utilized to test the bacterial susceptibility of the strains. This study explored the types and community structure of some culturable microorganisms in the intestinal tract of G. gratiosa and recorded their physiological characteristics. These data reflect the physiological functions of the intestinal microorganisms of G. gratiosa and provide support for subsequent research on the interaction mechanism between microorganisms and their hosts.

Habitat destruction and fragmentation are among the major current threats to global biodiversity. Fragmentation may also affect species with good dispersal abilities. We study the heath bushcricket Gampsocleis glabra, a specialist of steppe-like habitats across Europe that are highly fragmented, investigating if these isolated populations can be distinguished using population genomics and if there are any traces of admixture or dispersal among them. We try to answer these questions using genome-wide SNP data generated with ddRAD sequencing. We calculated F-statistics and visualized differentiation using STRUCTURE plots. While limited by the difficulty of sampling this threatened species, our results show that all populations except one that was represented by a singleton were clearly distinct, with pairwise FST values between 0.010 and 0.181. STRUCTURE indicated limited but visible admixture across most populations and probably also an exchange of individuals between populations of Germany and The Netherlands. We conclude that in G. glabra, a certain amount of gene flow has persisted, at least in the past, also among populations that are isolated today. We also detect a possibly more recent dispersal event between a population in The Netherlands and one in Germany, which may be human aided. We suggest that the conservation of larger populations should be maintained, that efforts should be taken to restore abandoned habitat, that the preservation even of small habitat fragments may be beneficial for the conservation of this species, and that these habitats should be regularly monitored for possible (re-)colonization.

Many predators and parasites eavesdrop on the communication signals of their prey. Eavesdropping is typically studied as dyadic predator–prey species interactions; yet in nature, most predators target multiple prey species and most prey must evade multiple predator species. The impact of predator communities on prey signal evolution is not well understood. Predators could converge in their preferences for conspicuous signal properties, generating competition among predators and natural selection on particular prey signal features. Alternatively, predator species could vary in their preferences for prey signal properties, resulting in sensory-based niche partitioning of prey resources. In the Neotropics, many substrate-gleaning bats use the mate-attraction songs of male katydids to locate them as prey. We studied mechanisms of niche partitioning in four substrate-gleaning bat species and found they are similar in morphology, echolocation signal design and prey-handling ability, but each species preferred different acoustic features of male song in 12 sympatric katydid species. This divergence in predator preference probably contributes to the coexistence of many substrate-gleaning bat species in the Neotropics, and the substantial diversity in the mate-attraction signals of katydids. Our results provide insight into how multiple eavesdropping predator species might influence prey signal evolution through sensory-based niche partitioning.