Gut flora alterations among aquatic firefly Aquatica leii inhabiting various dissolved oxygen in fresh water.

Knowledge about the impact of different dissolved oxygen (DO) on the composition and function of gut bacteria of aquatic insects is largely unknown. Herein, we constructed freshwater environments with different DOs (hypoxia: 2.50 ± 0.50, normoxia: 7.00 ± 0.50, and hyperoxia: 13.00 ± 0.50 mg/L) where aquatic firefly Aquatica leii larvae lived for three months. Their gut flora was analyzed using the combination of 16S rRNA amplicon sequencing and metagenomics. The results showed no difference in alpha diversity of the gut flora between A. leii inhabiting various DOs. However, the relative abundance of several bacterial lineages presented significant changes, such as Pseudomonas. In addition, bacterial genes with an altered relative abundance in response to various DOs were primarily related to metabolism. The alteration of these functions correlated with the DO change. This is the first to uncover structure of gut flora under various DOs in aquatic insect larvae.

Gut microbial diversity and function analysis of the final-instar larvae of Protohermes xanthodes (Megaloptera: Corydalidae).

Protohermes xanthodes (Megaloptera, Corydalidae, Protohermes), widespread species in China, is an important insects treated as food and medicine for aquaculture. In this study, the gut microbiota was investigated by 16S rRNA microbial profiling. A total of 600 Amplicon Sequence Variants (ASV) were identified, Proteobacteria and Firmicutes were the main dominant phyla, and 27 genera ran through the entire digestive tract, mainly Hafnia-Obesumbacterium (Proteobacteria), Lactobacillus and Lactococcus (Firmicutes). The PICRUSt2 functional prediction of gut microbiota showed that the foregut abundant pathways related to metabolism, environmental information processing, and genetic information processing. while the midgut had the most abundant metabolic and environmental information processing pathways, including the prominent phosphotransferase system (PTS), propionate metabolism, and ß-lactam resistance. The hindgut had the weakest metabolic function, but its genetic information processing was more abundant than the foregut and midgut. Additionally, 26 strains of bacteria were isolated from the midgut microorganisms, with Firmicutes being the dominant bacteria, and some of the purified bacteria had potential probiotic and anti-pathogen functions. These findings suggest that there are differences in the microorganisms of the different gut floras of the larvae, and each flora has specific metabolic functions. This research could be used to further understand the function of gut microorganisms, explore the co-evolution of P. xanthodes and gut microorganisms, and promote healthy breeding based on gut microorganisms.

A new species of the genus Burmadactylus Heads, 2009 from mid-Cretaceous amber in north Myanmar (Orthoptera: Caelifera: Tridactyloidea).

This paper describes a new species of the genus Burmadactylus Heads, 2009 (Tridactylidae: Dentridactylinae) from Burmese amber, namely: Burmadactylus tenuicerci sp. nov. This new species is similar to Burmadactylus grimaldi Heads, 2009, but differs from latter by mesotibia basally inflated and almost as long as mesofemur; the second segment of cercus distinctly slender; paraproctal lobe covered with sparse and slender setae, one of the setae near the apex distinct thick and long.

Adults of Alderflies, Fishflies, and Dobsonflies (Megaloptera) Expel Meconial Fluid When Disturbed.

Chemical secretions are an effective means by which insects may deter potential enemies, already being studied extensively with regard to their chemicals, synthesis, toxicity, and functions. However, these defensive secretions have been little studied in Megaloptera. Herein, the fluid expelling behavior of adult alderflies (Sialidae), fishflies (Corydalidae: Chauliodinae), and dobsonflies (Corydalidae: Corydalinae), all of the order Megaloptera, is described in detail regarding the timing and possible function of this behavior. When disturbed artificially, both males and females could expel fluid from the anus. However, the frequency of expelling was much lower in alderflies than in fishflies and dobsonflies. The amount of expelled fluid relative to body weight was also smaller in alderflies. In fishflies and dobsonflies, the amount of expelled fluid decreased with adult age, probably because the fluid is little replenished once expelled. The cream-colored fluid seems to be meconial fluid produced via the Malpighian tubules at the pupal stage, which is usually discharged at adult emergence in most other holometabolous insects. However, adult fishflies and dobsonflies often expel it vigorously by bending their abdomen when disturbed after emergence. Thus, the fluid expelling may be an anti-predatory behavior, particularly in younger adults that can expel a relatively large amount of fluid.

A new genus and a new species of pygmy mole cricket from mid-Cretaceous amber in North Myanmar (Orthoptera: Caelifera: Tridactyloidea).

This paper describes a new genus with a new species from Burmese amber, namely: Latedactylus longapedi gen. et sp. nov. (Tridactylidae: Tridactylinae). L. longapedi gen. et sp. nov. is erected based on the cylindrical second segment of cercus longer and distinctly slender than the first; metatibia without swimming plates; metatarsus about three times as long as apical spurs of metatibia.

Two New Species of Ripipterygidae (Orthoptera, Tridactyloidea) from Mid-Cretaceous of Myanmar with a Key to the Genera of Tridactyloidea in Amber.

The abundance of insects in Burmese amber illustrates a highly diverse orthoptera community of the mid-Cretaceous, but the records of ripipterygids are relatively rare. Here, we reviewed the genus of Magnidactylus (Xu, Fang and Jarzembowski, 2020) and transfered it from Tridactylidae to Ripipterygidae. Based on four ambers specimens collected from northern Myanmar, two new species, Magnidactylusmirus sp. nov. and Magnidactylusgracilis sp. nov., wereerected. M.mirus sp. nov. can be characterized by its basal segment and apical segment of paraproctal lobes, which are equally thick and clavate. M.gracilis sp. nov. can be characterized by its apical segment of paraproctal lobes, which are distinctly swollen. Additionally, in order to facilitate the classification of amber specimens of Tridactyloidea, a key to the genera ofambers in this superfamily is provided.

Physiological Responses of the Firefly Pyrocoelia analis (Coleoptera: Lampyridae) to an Environmental Residue From Chemical Pesticide Imidacloprid.

Imidacloprid, a neonicotinoid insecticide, is widely applied to control insect pests across a broad spectrum. Though the impact of residues from this chemical pesticide on non-target organisms in the field has been reported, it was not well characterized across a wide range of ecosystems, especially for some species considered as environmental indicators that live in forests. The effects of sublethal dose of imidacloprid on firefly, Pyrocoelia analis, were analyzed physiologically and biochemically in this study to better understand the impact of chemical pesticide application on environmental indicators such as fireflies. After imidacloprid treatment, the midgut tissues of the larva presented an abnormal morphology featured as atrophy of fat body cells, shrinking cells, and the destruction of a midgut structure. The activities of antioxidant enzymes, superoxide dismutase, catalase, and peroxidase were noticeably increased during early exposure to sublethal imidacloprid and then decreased at later stages. The malondialdehyde content significantly increased after 12 h of exposure to imidacloprid compared with the control. Similarly, the enzyme activities of polyphenol oxidase and acetylcholinesterase were increased after the imidacloprid treatment and then decreased at the later stage. In summary, a sublethal dose of imidacloprid caused destructive change in the tissue structure, and this damage was followed by an excessive reactive oxygen species that could not be eliminated by antioxidant enzymes. Our results indicated that the residues of imidacloprid might cause severe toxicity to non-target insects in the environment even far away from the agro-ecosystem where the chemicals were applied.

A Chromosome-Level Genome Assembly of the Pygmy Mole Cricket Xya riparia.

The pygmy mole cricket Xya riparia (Orthoptera: Tridactyloidea) is rarely studied or widely known. Some species of pygmy mole crickets, however, not only have a potential ecological value but are also important in the study of the evolution of the orthopteran genome and its phylogenetic relationships. The genome resources of pygmy crickets are limited and there are currently no publications referencing this species' genome. In this study, we assembled a reference genome of X. riparia at the chromosomal level using nanopore sequencing and Hi-C technology. An X. riparia genome of 1.67 Gb was successfully assembled from 164.01 Gb of nanopore sequencing data. The genome assembly showed a completeness of 98.97% benchmarking universal single-copy orthologs with a contig N50 of 4.18 Mb and the longest contig being 18.84 Mb. The contigs were clustered, ordered, and correctly oriented on six pseuchromosomes, which covered 95.63% of the genome assembly through Hi-C data with a scaffold N50 of 319.1 Mb and the longest scaffold being 397.8 Mb. Repeat sequences accounted for 42.88% of the whole-genome assembly. A total of 60,847 noncoding RNAs were detected. Moreover, 16,468 (87.91%) of the genes were functionally annotated. As this is the first high-quality reference genome of X. riparia at the chromosomal level, it will undoubtedly serve as a valuable resource for ecological, biological, and genetic research on pygmy mole crickets as well as for general research on Orthoptera's genome evolution and phylogenetic relationships.

Do predators prefer toxic animals? A case of chemical discrimination by an Asian snake that sequesters firefly toxins.

Several Asian natricine snakes of the genus Rhabdophis feed on toads and sequester steroidal cardiac toxins known as bufadienolides (BDs) from them. A recent study revealed that species of the Rhabdophis nuchalis Group ingest lampyrine fireflies to sequester BDs. Although several species of fireflies are distributed in the habitat of the R. nuchalis Group, only lampyrine fireflies, which have BDs, are included in the diet of these snakes. Thus, we hypothesized that the R. nuchalis Group chemically distinguishes fireflies that have BDs from those that do not have BDs. We also predicted that the R. nuchalis Group detects BDs as the chemical cue of toxin source. To test these predictions, we conducted 3 behavioral experiments using Rhabdophis chiwen, which belongs to the R. nuchalis Group. In the first experiment, R. chiwen showed a moderate tongue flicking response to cinobufagin, a compound of BDs. On the other hand, the snake showed a higher response to the chemical stimuli of lampyrine fireflies (BD fireflies) than those of lucioline fireflies (non-BD fireflies). In the second experiment, in which we provided live BD and non-BD fireflies, the snake voluntarily consumed only the former. In the third, a Y-maze experiment, the snake tended to select the chemical trail of BD fireflies more frequently than that of non-BD fireflies. These results demonstrated that R. chiwen discriminates BD fireflies from non-BD fireflies, but the prediction that BDs are involved in this discrimination was not fully supported. To identify the proximate mechanisms of the recognition of novel toxic prey in the R. nuchalis Group, further investigation is necessary.

Checklist and keys to Deltocephalinae leafhoppers (Hemiptera, Cicadellidae) from Pakistan.

Keys to all levels of the subfamily Deltocephalinae (Hemiptera: Cicadellidae) of Pakistan are provided based on published records and original data from recent research. Checklists to the genera and species of Deltocephalinae are also given. A total of 49 genera with more than 100 species are now known from Pakistan. Two new synonyms are proposed, i.e., Cicadulinastriata Ahmed, 1986 a junior synonym of Cicadulinachinai Ghauri, 1965, syn. nov. and Macrostelesparafalcatus Naveed & Zhang, 2018 a new junior synonym of Macrostelesindrina (Pruthi, 1930), syn. nov.

First report of the Old World leafhopper genus Olidiana McKamey (Hemiptera: Cicadellidae: Coelidiinae) from Pakistan with description of a new species.

The genus Olidiana McKamey, 2006 is reported for the first time from Pakistan based on the new species Olidiana pakistanica sp. nov. Photographic illustrations and a description are provided for the habitus and male genitalia of this new species. A partial mitochondrial cytochrome oxidase I gene (COI) sequence is also provided.

Two new species of the genus Xya Latreille, 1809 (Orthoptera, Tridactyloidea, Tridactylidae) from Yunnan with a key to all Xya species in China.

This contribution to the taxonomy of Xya Latreille, 1809 (Orthoptera, Tridactyloidea, Tridactylidae) adds descriptions and photographic illustrations of two new species: Xya xishangbanna sp. nov. and Xya yunnanensis sp. nov. from Xishuangbanna, Yunnan Province, China. Xya xishangbanna sp. nov. can be diagnosed by the shiny dark brown hind femora, and the epiproct with a shallow bottom of the middle "v-shaped" crack in the upper part and straight sides; Xya yunnanensis sp. nov. can be diagnosed by the compound eye bearing no narrow band along the inner margin, and the epiproct with the bottom of the side edge with a sharply angled protrusion and a narrow lower anchor-shaped base less than 1/2 the width of the upper one. Distributional information and bionomics for these two new species and photos for the habitat are given. A key to all Chinese species of Xya is provided.

A new species of the genus Xya Latreille, 1809 from Fujian, China (Orthoptera, Tridctyloidea, Tridactylidae).

The genus Xya Latreille, 1808 obtains 58 known species in the world, among them 18 species distributed in Asia and 8 species in China only i.e. Xya japonica (Haan, 1842), Xya riparia (Saussure, 1877), Xya nitobei (Shiraki, 1911), Xya apicicornis (Chopard, 1928), Xya manchurei Shiraki, 1936, Xya lehsanensis Cao et al, 2017, Xya sichuanensis Cao et al, 2018, Xya shandongensis, Zhang et al, 2018; [Latreille, 1809; Haan, 1844; Walker, 1871; Saussure, 1877, 1896; Brunner von Wattenwyl. 1893; Bolivar, 1900(1899); Shiraki,1911, 1936; Chopard, 1920, 1928, 1936, 1968; Tindale, 1928; Willemse, 1954; Bey-Bienko, 1967; Harz, 1970, 1971; Günther, 1974, 1978, 1980, 1982, 1990, 1995, 1998; Ingrisch, 1987, 2006; Baehr, 1988; Yin et al, 1996; Murai, 2005; Yin et al, 2013; Heads Hollier, 2016; Kuravova Kocarek, 2016; Cao et al, 2017; Cao et al, 2018; Zhang et al, 2018; Cigliano, et al, 2019]. In the present paper, a new species of the genus Xya Latreille, 1809 from Fujian, China is described. The new species Xya fujianensis sp. nov. is similar to Xya lehsanensis Cao et al, 2017, but differs from the latter by head black, with a yellow band along inner margin of eye; pronotum black, with a white band on the lower margin; fore wing black, with two yellow spots near base and two yellow spots near top and hind femur black with two large yellow spots on upper side. Type specimens are deposited in the College of Life Science, Leshan Normal University, Leshan 614004, China.

Dramatic dietary shift maintains sequestered toxins in chemically defended snakes.

Unlike other snakes, most species of Rhabdophis possess glands in their dorsal skin, sometimes limited to the neck, known as nucho-dorsal and nuchal glands, respectively. Those glands contain powerful cardiotonic steroids known as bufadienolides, which can be deployed as a defense against predators. Bufadienolides otherwise occur only in toads (Bufonidae) and some fireflies (Lampyrinae), which are known or believed to synthesize the toxins. The ancestral diet of Rhabdophis consists of anuran amphibians, and we have shown previously that the bufadienolide toxins of frog-eating species are sequestered from toads consumed as prey. However, one derived clade, the Rhabdophis nuchalis Group, has shifted its primary diet from frogs to earthworms. Here we confirm that the worm-eating snakes possess bufadienolides in their nucho-dorsal glands, although the worms themselves lack such toxins. In addition, we show that the bufadienolides of R. nuchalis Group species are obtained primarily from fireflies. Although few snakes feed on insects, we document through feeding experiments, chemosensory preference tests, and gut contents that lampyrine firefly larvae are regularly consumed by these snakes. Furthermore, members of the R. nuchalis Group contain compounds that resemble the distinctive bufadienolides of fireflies, but not those of toads, in stereochemistry, glycosylation, acetylation, and molecular weight. Thus, the evolutionary shift in primary prey among members of the R. nuchalis Group has been accompanied by a dramatic shift in the source of the species' sequestered defensive toxins.

Allometry and morphological trait relationship in the sexually dimorphic Chinese dobsonfly, Acanthacorydalisasiatica (Wood-Mason, 1884) (Megaloptera, Corydalidae).

Male insects with large weapons such as horns and elongate mandibles would be expected to invest more on such structures than other parts of the body for advantages in male to male competition for mating. In male genitalia, however, intermediate size provides a better fit for more females than small or large sizes, and such a male would leave more offspring regardless of their body size. These predictions were tested using a static allometry analysis between body size and other trait sizes. Acanthacorydalisasiatica is a large dobsonfly (Megalotera) and males have conspicuously large mandibles used as weapons. We examined the hypothesis that the male mandibles of this sexually dimorphic species are sexually selected to enlarge, whereas the male genitalia are stable to be intermediate regardless of a great variation in body size. The results, as predicted, showed positive allometry between male body size and mandible length but negative allometry between male body size and ectoproct length (a male grasping structure). Sperm are transferred through a small spermatophore attached externally to the female genital opening, so it may be evolutionarily unnecessary to develop an enlarged male genital size. In contrast, there may be a trade-off between male mandible size and wing length, because of negative allometry between body size and wing length in males but isometry between them in females.

A new species and key to all known species of the genus Ceracris Walker, 1870 from China (Orthoptera: Acridoidea, Arcypteridae).

The genus Ceracris Walker, 1870 is a median genus in Arcypteridae, which contains 23 species in the world, among them 11 species distributed in China [Walker, 1870; Bolivar, 1914; Uvarov, 1931; Zheng, 1977; Liu, 1981; Zheng, 1983; Yin, 1984; Liang, 1988; Wang, 1992; Liu et Li, 1995; Ou et al, 1995; Yin et al, 1996; Zheng et Xia, 1998; Zhang et al, 2017; Feng et al, 2018; Cigliano, 2018]. A new species of the genus from Hainan, China is described in this paper. The new species i.e. Ceracris amplicornis sp. nov. is similar to Ceracris versicolor (Brunner von Wattenwyl, 1893), but differs from latter by antennae short, length of a segment in the middle 2.9 times its width; lateral carinae of pronotum not intact and indistinct in metazona; median area of tegmen without intercalary vein; bridge of epiphallus straight and tegmen of female short, reaching the end of hind femur only. Type specimens are deposited in the Natural Museum of Hebei University, Baoding, Hebei, China.

A new genus and a new species of pygmy mole cricket in Cretaceous amber from Burma (Orthoptera: Tridactylidae).

A new genus and a new species of pygmy mole cricket i.e. Paraxya gen nov. and Paraxya hui sp. nov. are described from Burma Cretaceous amber in this paper. The new genus is allied to Xya Latreille. 1809, but differs from latter by lacking swimming plates at metatibia [Latreille. 1809; Yin et al, 1996; Cao et al, 2017, Cao et al, 2018 and Zhang et al, 2018]. The new genus is also allied to Burmadactylus Heads, 2009, but differs from latter by metatarsus absent and hindwing present, surpassing the end of abdomen distinctly [Heads, 2009]. Type specimen is deposited in the College of Life Science, Leshan Normal University, Leshan 614004, China.

Description of the final-instar larva and pupa of Neoneuromus ignobilis Navás, 1932 (Megaloptera: Corydalidae).

The immature stages of only one species of the dobsonfly genus Neoneuromus van der Weele are known. The final-instar larva and pupa of N. ignobilis Navás, 1932, the most common and widespread species of the genus, is herein described for the first time. Several morphological features of the larvae were found to distinguish N. ignobilis from other dobsonfly species with similar large-sized mature larvae.