An integrative approach to the study of Kempnyia Klapálek, 1914 (Plecoptera: Perlidae) from Brazil: Support for the description of four new species and a basis for future studies.

Kempnyia (Plecoptera: Perlidae) is an endemic genus of Brazilian stoneflies that has 36 valid species and is distributed primarily in the Atlantic Forest and the mountainous areas of Central Brazil, particularly in Goiás and Tocantins states. Despite being the Brazilian genus with the most DNA sequences available on GenBank, integrative studies on the genus began only recently, in 2014. In this context, herein we studied the morphology and molecular data of Kempnyia specimens deposited in the Aquatic Biology Laboratory (UNESP, Assis) and the Entomology Museum of the Federal University of Viçosa (UFVB, Viçosa) collections. For the integrative approach adopted, in addition to studying the specimens morphologically, we used sequences of the COI mitochondrial gene combined with the following species delimitation methods: Automatic Barcode Gap Discovery (ABGD), both primary (ABGDp) and recursive (ABGDr) partitions; Assemble Species by Automatic Partitioning (ASAP); Poisson Tree Processes (PTP) and the Bayesian implementation of the Poisson Tree Processes (bPTP). As a result, we provided 28 new COI sequences of 21 species and support the description of four new species, namely, K. guarani sp. nov., K. tupiniquim sp. nov., K. una sp. nov., and K. zwickii sp. nov., consequently increasing the known diversity of the genus to 40 species. We also discuss the morphological variations observed in other species of the genus and provide several new geographic records. Therefore, our study brings new insights into the values of intra- and interspecific molecular divergence within Kempnyia, serving as a basis for new studies.

The dark side of organic farming: Copper sulphate compromises the life history and behaviour of the walking stick insect, Bacillus rossius.

Organic farming is considered the most sustainable form of modern soil cultivation. Yet it often relies on the use of chemical compounds that are not necessarily harmless for the surrounding wildlife. In this study, we tested the effects of realistic concentrations of copper sulphate-largely used in organic farming as a fungicide-on ecologically-relevant traits of the walking stick insect Bacillus rossius, a species commonly found in the proximity of cultivated fields across Europe. By using second-generation progeny of wild-caught parthenogenetic females bred in common gardens, we measured the impact of copper sulphate (CuSO4) on both the life-history (body condition, number of eggs, and hatching success) and behavioural traits (activity and maximum vertical speed) of the individuals. We observed strong negative effects of high, realistic concentrations of copper sulphate on most traits within 12 days of exposure, while effects were less evident at lower concentrations of the pollutant. Our results reveal that realistic concentrations of copper sulphate can compromise important traits that regulate both the survival and reproduction of animals in the wild, with such effects that are, however, dose dependent. We suggest that common practices in organic farming require further consideration on their ecological and evolutionary impact on wildlife.

Morphology of the pterothoracic musculature in Paraneoptera and its phylogenetic implication (Insecta: Neoptera).

Although the monophyly of Paraneoptera (=hemipteroid orders or Acercaria, composed of Psocodea, Thysanoptera and Hemiptera) has been widely accepted morphologically, the results from molecular phylogenetic and phylogenomic analyses contradict this hypothesis. In particular, phylogenomic analyses provide strong bootstrap support for the sister group relationship between Psocodea and Holometabola, that is, paraphyly of Paraneoptera. Here, we examined the pterothoracic musculature of Paraneoptera, as well as a wide range of other neopterous insect orders, and analysed its phylogenetic implication. By using the synchrotron microcomputed tomography (µCT) and parsimony-based ancestral state reconstruction, several apomorphic conditions suggesting the monophyly of Paraneoptera, such as the absence of the II/IIItpm7, IIscm3, IIIspm2 and IIIscm3 muscles, were identified. In contrast, no characters supporting Psocodea + Holometabola were recovered from the thoracic muscles. These results provide additional support for the monophyly of Paraneoptera, together with the previously detected morphological apomorphies of the head, wing base, and abdomen.

The chromosome-level genome assembly of the giant dobsonfly Acanthacorydalis orientalis (McLachlan, 1899).

Acanthacorydalis orientalis (McLachlan, 1899) (Megaloptera: Corydalidae) is an important freshwater-benthic invertebrate species that serves as an indicator for water-quality biomonitoring and is valuable for conservation from East Asia. Here, a high-quality reference genome for A. orientalis was constructed using Oxford Nanopore sequencing and High throughput Chromosome Conformation Capture (Hi-C) technology. The final genome size is 547.98 Mb, with the N50 values of contig and scaffold being 7.77 Mb and 50.53 Mb, respectively. The longest contig and scaffold are 20.57 Mb and 62.26 Mb in length, respectively. There are 99.75% contigs anchored onto 13 pseudo-chromosomes. Benchmarking Universal Single-Copy Orthologs (BUSCO) analysis showed that the completeness of the genome assembly is 99.01%. There are 10,977 protein-coding genes identified, of which 84.00% are functionally annotated. The genome contains 44.86% repeat sequences. This high-quality genome provides substantial data for future studies on population genetics, aquatic adaptation, and evolution of Megaloptera and other related insect groups.

Sensory pathway in aquatic basal polyneoptera: Antennal sensilla and brain morphology in stoneflies.

Aquatic insects represent a great portion of Arthropod diversity and the major fauna in inland waters. The sensory biology and neuroanatomy of these insects are, however, poorly investigated. This research aims to describe the antennal sensilla of nymphs of the stonefly Dinocras cephalotes using scanning electron microscopy and comparing them with the adult sensilla. Besides, central antennal pathways in nymphs and adults are investigated by neuron mass-tracing with tetramethylrhodamine, and their brain structures are visualized with an anti-synapsin antibody. No dramatic changes occur in the antennal sensilla during nymphal development, while antennal sensilla profoundly change from nymphs to adults when switching from an aquatic to an aerial lifestyle. However, similar brain structures are used in nymphs and adults to process diverging sensory information, perceived through different sensilla in water and air. These data provide valuable insights into the evolution of aquatic heterometabolous insects, maintaining a functional sensory system throughout development, including a distinct adaptation of the peripheral olfactory systems during the transition from detection of water-soluble chemicals to volatile compounds in the air. From a conservation biology perspective, the present data contribute to a better knowledge of the biology of stoneflies, which are very important bioindicators in rivers.

A new species of the genus Embia Latreille, 1825 (Insecta, Embioptera) from the Madonie Regional Natural Park (Sicily, Italy).

Embia minapalumboi n. sp., a new species of the genus Embia Latreille, 1925, was collected during an excursion in the Madonie Regional Natural Park that took place at the conclusion of the XXVII Italian National Congress of Entomology celebrated in Palermo from 12 to 16 June 2023. In the location of discovery, at an elevation of just under 1400 m and therefore unusual for European Embioptera, the new species was found to be very rare and for this reason most of the few juvenile specimens found were kept in breeding by the author. The new species was compared with all 36 species known to date for the genus and in particular with the 21 species from the Mediterranean area and the 8 known from Europe. The author also presents the general distribution of all species of the genus. Embia tyrrhenica, Stefani, 1953 is also reported for the Madonie and reconfirmed for Sicily.

Description of Neoperla philippinensis sp. nov. confirmed using integrative taxonomy and some notes on Neoperla oculata Banks, 1924 (Plecoptera: Perlidae).

I re-examine the assumed phenotypic variable Neoperla oculata Banks, 1924 (Plecoptera, Perlidae) and describe Neoperla philippinensis sp. nov. All material examined herein are collections from the Mindanao region, of which supplementary information were provided including COI mtDNA accessions, female adult associations, and egg morphology characterization.

A new species and three records of Chloroperlidae (Plecoptera) from China.

A new species of the genus Suwallia Ricker, 1943, S. rostrata sp. nov., from Xizang of southwestern China is described, illustrated and compared with similar species. Alloperla kurentzovi Zhiltzova & Zapekina-Dulkeit, 1977, A. kurilensis Zhiltzova, 1978, A. teleckojensis mal, 1939 are reported for the first time from China. The diagnostic features of these species are illustrated with color images.

Dr. Wolfram Meys Stonefly (Insecta: Plecoptera) Collection from Luzon, Philippines: Part I. Perlidae.

In this study, the Perlidae stonefly collections of Dr. Wolfram Mey from over 20 years ago have been examined. Neoperla jhoanae sp. nov. from Central Luzon Region and Neoperla nagaensis sp. nov. from Bicol Region are described herein. Additional records and information are also provided on Neoperla jewetti Sivec, 1984, Neoperla recta Banks, 1913, Neoperla pseudorecta Sivec, 1984, Neoperla oculata Banks, 1924, Neoperla nigra Sivec, 1984, Phanoperla bakeri (Banks, 1924), and Phanoperla flaveola (Klaplek, 1910).

Additional records of Neoperla (Plecoptera: Perlidae) from Yunnan Province of China, with description of a new species.

A new species of the genus Neoperla, N. donglaii sp. nov., is described from Yunnan Province of southwestern China. The new species is assigned to the N. lushana-complex of the N. (Formosita) subgenus due to the presence of paired, ventral spiny lobes of the aedeagus. The new species was compared with related species in the subgenus. Neoperla hamata Jewett, 1975 and N. idella Stark & Sivec, 2008 are newly recorded for China and N. quadrata Wu & Claassen, 1934 specifically from Yunnan Province, China. Neoperla quadrata had not been collected for 90 years.

Revision of the bark louse genus Bicuspidatus (Psocodea: Peripsocidae) with description of a new species from China.

Bicuspidatus is a small genus of the family Peripsocidae, erected by Li (1993) with B. pulchipunctatus as the type species. Adults of this genus can be easily distinguished from other peripsocids by the phallosome with bicuspid apical process in males or by the subgenital plate with modestly elongated, distally bilobed posterior lobe in females. Bicuspidatus currently comprises three species: B. guangdongensis Li, 1993, B. pulchipunctatus Li, 1993 and B. sigillatus Li, 1993, all are known only from Guangdong Province, China. According to previous studies, these three species can be distinguished by the markings on the forewings and a few characteristics of phallosome (Li 1993, 2002). In this study, we present a taxonomic revision of Bicuspidatus based on examination of 40 specimens collected from various localities, including the type specimens of B. guangdongensis, B. pulchipunctatus and B. sigillatus. Furthermore, 12 of them were used for molecular analyses based on partial sequences of COI gene. As a result, B. guangdongensis and B. sigillatus are here considered as junior synonyms of B. pulchipunctatus. We redescribe B. pulchipunctatus and describe a new species from Xizang Autonomous Region, China. An updated distribution map and a revised key to the species of Bicuspidatus are also provided.

Identity on Filchneria heteroptera (Wu, 1938) (Plecoptera: Perlodidae) with a new record of Paraleuctra cercia (Okamoto, 1922) from China.

Filchneria heteroptera (Wu, 1938) is rediscovered from Heilongjiang Province of northeastern China. A supplementary description and illustrations including aedeagal structures based on newly collected specimens are provided. Filchneria dongruihangi Chen, 2020 is considered here as a junior synonym of F. heteroptera (Wu, 1938). Paraleuctra cercia (Okamoto, 1922) is reported from Liaoning and Heilongjiang provinces, China for the first time.

A remarkable new species and one newly recorded species of genus Oxyartes Stl, 1875 (Phasmida, Lonchodidae, Necrosciinae) from China.

The genus Oxyartes currently comprises 17 taxa, of which 12 are found in China, representing the high diversity in China. This list includes the two species from China as described in this paper. The first is a new remarkable species O. bouxraeuz sp. nov. collected from Gulinqing township, Yunnan. The second is a newly recorded species, O. cresphontes. This species is reported from Mdog, Xizang, China. A key to this genus from China is presented. Type specimens are deposited in the Yunnan Agricultural University (YNAU).

The only species of Calvisia (Phasmatodea, Necrosciinae) from China, with a new synonymy and additional descriptions.

Calvisia is a colorful winged stick insect genus consisting of 6 subgenera and 44 species widely distributed in temperate and tropical Asia. C. medogensis syn. nov. was discovered in Mdog, Xizang (Tibet), China and is so far the only species recorded from China. We here propose that C. medogensis syn. nov. is a synonym of C. fuscoalata after checking type specimens of both species. New materials studied are deposited in Yunnan Agricultural University, China (YNAU).

A new species of Jeremia Redtenbacher (Phasmatodea: Phasmatidae: Cladomorphinae), a stick insect from the northern Brazilian Amazon Basin.

A new species of Jeremia Redtenbacher (Phasmatodea: Phasmatidae: Cladomorphinae) is described based on both male and female specimens from the northern Brazilian Amazon Basin and named Jeremia megaplax sp. nov. The new species can be differentiated from other species of this genus by the length of the female subgenital plate, which is two times longer than the length of tergites 810. Photos and description from subadult male and not fully developed eggs are also provided. The studied material is deposited at the Invertebrate Collection of the Instituto Nacional de Pesquisas da Amaznia (INPA).

Synopsis of the Ptiloneuropsis Roesler, 1940 (Psocodea: Psocoptera: Ptiloneuridae), with a new cave-dwelling species from Brazil.

A new cave-dwelling species of Ptiloneuropsis from southeastern Brazil, is here described based on both sexes, illustrated and named as: Ptiloneuropsis krenaki sp. nov. This new species differs from Ptiloneuropsis immaculata Roesler, 1940 and Ptiloneuropsis diamantina Silva-Neto, Garca Aldrete & Rafael, 2018 in details of the hypandrium and phallosome. In addition to the third new species of the Ptiloneuropsis, descriptions of variation in the fore and hindwings veins are also given. The diagnosis of Ptiloneuropsis is updated, and P. immaculata is redescribed. Furthermore, the first identification key for Ptiloneuropsis species, based on males, is presented.

Notes on the genus Dictyogenus Klaplek, 1904 (Plecoptera, Perlodidae) in the Austrian, Italian, Slovenian and Swiss Alps, with the description of six new species.

Six new species of Dictyogenus Klaplek, 1904, from the Austrian, Italian, Slovenian, and Swiss Alps, are described and illustrated from adults and nymphs. The species Dictyogenus alpinum (Pictet, 1841) and topotypical Dictyogenus fontium (Ris, 1896) are re-described.

Mitochondrial genomes provide insights into the Euholognatha (Insecta: Plecoptera).

BACKGROUND: Euholognatha is a monophyletic group within stoneflies comprised by a superfamily Nemouroidea and a family Scopuridae. Based on morphological data, the family-level phylogenetic relationships within Euholognatha are widely accepted, but there is still controversy among different molecular studies. To better understand the phylogeny of all six extant euholognathan families, we sequenced and analyzed seven euholognathan mitogenomes. RESULTS: The sequence heterogeneity analysis observed a low degree of compositional heterogeneity in euholognathan mitogenomes. Meanwhile, leuctrid mitogenomes were more heterogeneous than other euholognathan families, which may affect the phylogenetic reconstruction. Phylogenetic analyses with various datasets generated three topologies. The Leuctridae was recovered as the earliest branching lineage, and the sister relationship of Capniidae and Taeniopterygidae was supported by most tree topologies and FcLM analyses. When separately excluding sparsely sampled Scopuridae or high heterogeneity leuctrid taxa, phylogenetic analyses under the same methods generated more stable and consistent tree topologies. Finally, based on the results of this study, we reconstructed the relationships within Euholognatha as: Leuctridae + (Scopuridae + ((Taeniopterygidae + Capniidae) + (Nemouridae + Notonemouridae))). CONCLUSION: Our research shows the potential of data optimizing strategies in reconstructing phylogeny within Euholognatha and provides new insight into the phylogeny of this group.

Subcortical life, evolution of flattened body, and constrained mating posture in the earwig Platylabia major (Insecta: Dermaptera: "Anisolabididae").

Many animals take advantage of the shaded, humid, and protected environments in subcortical spaces, i.e., thin spaces under the loosened bark of dead trees. Permanent inhabitants of subcortical spaces often show specialized morphologies, such as a miniaturized or dorsoventrally flattened body. However, the evolutionary consequences of these specialized morphologies on behavioral, ecological, and life-history traits have been little studied. We studied the mating biology and anatomy of Platylabia major (usually placed in the family Anisolabididae), which is an obligate inhabitant of subcortical spaces with a paper-like flattened body, and compared them with those of two thicker, spongiphorid earwigs, Nesogaster amoenus and Paralabellula curvicauda. Mating trials in various settings showed that Pl. major requires thin spaces sandwiched by two planes to accomplish genital coupling and insemination. In contrast, the thicker species, although also frequently found in subcortical spaces, could mate on a single horizontal plane due to the ability of the male to twist its abdomen through approximately 180°. Examination by micro-computed tomography and a reagent-based clearing technique revealed no substantive differences in the configuration of mid-abdominal musculature between the species. The dorsal and lateral muscles of Pl. major, which are almost parallel to the antero-posterior body axis for accommodation within the thin abdomen, seemed incapable of producing the power to twist the abdomen. The abdominal musculature conforms to a simple pattern in both male and female earwigs, which is repeated in each of the pregenital segments. We conclude that small differences in the range of motion of each abdominal segment can result in large differences in possible mating postures and positions. Surgical experiments also demonstrated that both right and left penises of Pl. major are competent and used for insemination with no lateral bias, as in most other earwigs with twin penises studied to date.

Taxonomic Contributions on Macrogynoplax Enderlein and Enderleina Jewett (Plecoptera: Perlidae) from Brazil, Including a New Species Supported by Molecular and Morphological Data.

Stoneflies (Plecoptera) are aquatic insects widely used in ecological and monitoring studies as indicators of environmental quality due to their high sensitivity to pollution. Among Brazilian Plecoptera, Perlidae is the most diverse family, with four genera (Anacroneuria Klapálek 1909, Enderleina Jewett 1960, Kempnyia Klapálek 1914, and Macrogynoplax Enderlein 1909) and about 150 species. Herein, based on data from the barcode region of the mitochondrial cytochrome c oxidase subunit I (COI) gene and morphological characters, we studied specimens of Enderleina and Macrogynoplax, mainly from the Brazilian Savanna. Our alignment with a total length of 608 base pairs included 23 COI sequences, of which 10 are new and 13 were obtained from GenBank. The sequences and congruence of the obtained species clusters were analyzed using neighbor joining. As a result, we describe a new species of Enderleina, the first of this genus in the Brazilian Savanna, and update the geographic distribution of some Macrogynoplax species. A new record was included for Macrogynoplax veneranda Froehlich 1984 in São Paulo state and the first record of M. yupanqui Stark 1996 in Brazil was also included. First records were made for Macrogynoplax matogrossensis Bispo and Neves 2005 and a Macrogynoplax specimen in Goiás and Bahia states, respectively. Consequently, we expanded the distribution of the genus in the Brazilian Savanna and Atlantic Forest. In addition, we provide observations on the morphological differences found among these species, discuss about the distribution of the genera, present molecular data discussing their contributions to the knowledge of the genus, and comment on the recommendations for future research considering a scenario of integrative approaches. Lastly, we present a key to identify the adult males of Enderleina.