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Abstract Background Cryptolaemus montrouzieri (Coccinellidae) is widely utilized as biological control agents in modern agriculture. A comprehensive understanding of its food preference can help guide mass rearing and safety management during field application of pest control. Although some studies have paid attentions to the impacts of prey shift on C. montrouzieri, little is known regarding the role of post-transcriptional regulations in its acclimation to unnatural preys. Results We performed a genome-wide investigation on alternative splicing dynamics in C. montrouzieri in response to the predation transition from natural prey to unnatural ones. When feeding on undesired diets, 402–764 genes were differentially alternative spliced in C. montrouzieri. It is noteworthy that the majority of these genes (> 87%) were not differentially expressed, and these differentially spliced genes regulated distinct biological processes from differentially expressed genes, such as organ development and morphogenesis, locomotory behavior, and homeostasis processes. These suggested the functionally nonredendant role of alternative splicing in modulating physiological and metabolic responses of C. montrouzieri to the shift to undesired preys. In addition, the individuals feeding on aphids were subject to a lower level of changes in splicing than other alternative diets, which might be because of the similar chemical and microbial compositions. Our study further suggested a putative coupling of alternative splicing and nonsense-mediated decay (AS-NMD), which may play an important role in fine-tuning the protein repertoire of C. montrouzieri, and promoting its acclimation to predation changes. Conclusion These findings highlight the key role of alternative splicing in modulating the acclimation of ladybirds to prey shift and provide new genetic clues for the future application of ladybirds in biocontrol.

ABSTRACTIn this study, we first time sequenced and analyzed the 16S rRNA gene data of predator ladybird beetles Novius pumilus and globally distributed invasive pest Icerya aegyptiaca at different stages, and combined data with bacterial genome sequences in N. pumilus to explored the taxonomic distribution, alpha and beta diversity, differentially abundant bacteria, co-occurrence network, and putative functions of their microbial community. Our finding revealed that Candidatus Walczuchella, which exhibited a higher abundance in I. aegyptiaca, possessed several genes in essential amino acid biosynthesis and seemed to perform roles in providing nutrients to the host, similar to other obligate symbionts in scale insects. Lactococcus, Serratia, and Pseudomonas, more abundant in N. pumilus, were predicted to have genes related to hydrocarbon, fatty acids, and chitin degradation, which may assist their hosts in digesting the wax shell covering the scale insects. Notably, our result showed that Lactococcus had relatively higher abundances in adults and eggs compared to other stages in N. pumilus, indicating potential vertical transmission. Additionally, we found that Arsenophonus, known to influence sex ratios in whitefly and wasp, may also function in I. aegyptiaca, probably by influencing nutrient metabolism as it similarly had many genes corresponding to vitamin B and essential amino acid biosynthesis. Also, we observed a potential horizontal transfer of Arsenophonus between the scale insect and its predator, with a relatively high abundance in the ladybirds compared to other bacteria from the scale insects.IMPORTANCEThe composition and dynamic changes of microbiome in different developmental stages of ladybird beetles Novius pumilus with its prey Icerya aegyptiaca were detected. We found that Candidatus Walczuchella, abundant in I. aegyptiaca, probably provide nutrients to their host based on their amino acid biosynthesis-related genes. Abundant symbionts in N. pumilus, including Lactococcus, Serratia, and Pseudophonus, may help the host digest the scale insects with their hydrocarbon, fatty acid, and chitin degrading-related genes. A key endosymbiont Arsenophonus may play potential roles in the nutrient metabolisms and sex determination in I. aegyptiaca, and is possibly transferred from the scale insect to the predator.

The Proterozoic oil in Gaosheng area of the Western Depression of Liaohe Basin has great potential and is an important exploration field. However, the study about control factors of Gaosheng reservoir needs to be improved. The source rocks are characterized by using geochemical techniques and the structural evolution history is restored by using 2D-Move software. The characteristics of reservoirs and caps are clarified, and the controlling factors of reservoir formation are summarized. The oil generation intensity of Es4 and Es3 is generally 5 × 106–10 × 106 t/km2, indicating that the oil source is sufficient. The sandstone reservoir has fracture and dissolution porosity, which provide storage space for oil accumulation. Faults and unconformities provide pathways for oil migration. The dense Fangshenpao and Mesozoic basalt is a good cap layer to preserve the reservoir, which effectively prevents the upward migration of oil. According to the comprehensive analysis of main controlling factors, the favorable exploration areas are pointed out. The research results are of great significance to guide the future oil and gas exploration in Liaohe Basin.

The eggs of the Mediterranean flour moth, Ephestia kuehniella, are frequently utilized as alternative diets and have demonstrated promising outcomes when consumed by various insects. Nonetheless, the specific reasons for their effectiveness remain unclear. In our study, we assessed the developmental performance of the ladybird Propylea japonica when fed E. kuehniella eggs, alongside 12 factitious prey or artificial diets. Our findings revealed that ladybirds fed E. kuehniella eggs displayed a performance comparable to those fed the natural prey Megoura crassicauda. Transcriptome profiling of larvae raised on E. kuehniella eggs and M. crassicauda revealed that genes upregulated in the former group were enriched in metabolic pathways associated with carbohydrates, lipids, and other essential nutrients. This suggests that E. kuehniella eggs may have a higher nutrient content compared to natural prey. Furthermore, a notable downregulation in the expression of immune effector genes, such as Attacin and Coleoptericin, was observed, which might be attributed to the lower microbial content in E. kuehniella eggs compared to M. crassicauda. We suggest that the difference between E. kuehniella eggs and M. crassicauda as food sources for P. japonica lies in their nutrient and microbial contents. These findings provide valuable insights for the advancement of innovative artificial breeding systems for natural enemies.

The Shuguang area has great oil and gas potential in the Proterozoic and it is a major exploration target in the Western Depression. However, controlling factors and a reservoir-forming model of the Shuguang reservoir need further development. The characteristics of the reservoir formation in this area were discussed by means of a geochemical technique, and the controlling factors of the oil reservoir were summarized. The oil generation intensity of Es4 source rock was 25 × 106–500 × 106 t/km2, indicating that the source rocks could provide enough oil for the reservoir. The physical property of the quartz sandstone reservoir was improved by fractures and faults, which provided a good condition for the oil reservoir. Two periods of oil charging existed in the reservoir, with peaks of 38 Ma and 28 Ma, respectively. A continuous discharge of oil is favorable for oil accumulation. Oil could migrate through faults and fractures. In addition, the conditions of source–reservoir–cap assemblage in the Shuguang area well preserved the oil reservoir. The lower part of the Shuguang reservoir was source rock, the upper part was reservoir, and it was a structure-lithologic oil reservoir. These results are crucial for further oil exploration.

Abstract Background Scale insects are worldwide sap-sucking parasites, which can be distinguished into neococcoids and non-neococcoids. Neococcoids are monophyletic with a peculiar reproductive system, paternal genome elimination (PGE). Different with neococcoids, Iceryini, a tribe in non-neococcoids including several damaging pests, has abdominal spiracles, compound eyes in males, relatively abundant wax, unique hermaphrodite system, and specific symbionts. However, the current studies on the gene resources and genomic mechanism of scale insects are mainly limited in the neococcoids, and lacked of comparison in an evolution frame. Result We sequenced and de novo assembled a transcriptome of Icerya aegyptiaca (Douglas), a worldwide pest of Iceryini, and used it as representative of non-neococcoids to compare with the genomes or transcriptomes of other six species from different families of neococcoids. We found that the genes under positive selection or negative selection intensification (simplified as “selected genes” below) in I. aegyptiaca included those related to neurogenesis and development, especially eye development. Some genes related to fatty acid biosynthesis were unique in its transcriptome with relatively high expression and not detected in neococcoids. These results may indicate a potential link to the unique structures and abundant wax of I. aegyptiaca compared with neococcoids. Meanwhile, genes related to DNA repair, mitosis, spindle, cytokinesis and oogenesis, were included in the selected genes in I. aegyptiaca, which is possibly associated with cell division and germ cell formation of the hermaphrodite system. Chromatin-related process were enriched from selected genes in neococcoids, along with some mitosis-related genes also detected, which may be related to their unique PGE system. Moreover, in neococcoid species, male-biased genes tend to undergo negative selection relaxation under the PGE system. We also found that the candidate horizontally transferred genes (HTGs) in the scale insects mainly derived from bacteria and fungi. bioD and bioB, the two biotin-synthesizing HTGs were exclusively found in the scale insects and neococcoids, respectively, which possibly show potential demand changes in the symbiotic relationships. Conclusion Our study reports the first I. aegyptiaca transcriptome and provides preliminary insights for the genetic change of structures, reproductive systems and symbiont relationships at an evolutionary aspect. This will provide a basis for further research and control of scale insects.

Based on the analysis of Upper Paleozoic source rocks, source-reservoir-caprock assemblage, and gas accumulation characteristics in the Ordos Basin, the gas accumulation geological model of total petroleum system is determined. Then, taking the Carboniferous Benxi Formation and the Permian Taiyuan Formation and Shanxi Formation as examples, the main controlling factors of gas accumulation and enrichment are discussed, and the gas enrichment models of total petroleum system are established. The results show that the source rocks, faults and tight reservoirs and their mutual coupling relations control the distribution and enrichment of gas. Specifically, the distribution and hydrocarbon generation capacity of source rocks control the enrichment degree and distribution range of retained shale gas and tight gas in the source. The coupling between the hydrocarbon generation capacity of source rocks and the physical properties of tight reservoirs controls the distribution and sweet spot development of near-source tight gas in the basin center. The far-source tight gas in the basin margin is mainly controlled by the distribution of faults, and the distribution of inner-source, near-source and far-source gas is adjusted and reformed by faults. Generally, the Upper Paleozoic gas in the Ordos Basin is recognized in four enrichment models: inner-source coalbed gas and shale gas, inner-source tight sandstone gas, near-source tight gas, and far-source fault-transported gas. In the Ordos Basin, inner-source tight gas and near-source tight gas are the current focuses of exploration, and inner-source coalbed gas and shale gas and far-source gas will be important potential targets in the future.

Abstract Background Members of the genus Novius Mulsant, 1846 (= Rodolia Mulsant, 1850) (Coleoptera, Coccinellidae), play important roles in the biological control of cotton cushion scale pests, especially those belonging to Icerya. Since the best-known species, the vedalia beetle Novius cardinalis (Mulsant, 1850) was introduced into California from Australia, more than a century of successful use in classical biological control, some species of Novius have begun to exhibit some field adaptations to novel but related prey species. Despite their economic importance, relatively little is known about the underlying genetic adaptations associated with their feeding habits. Knowledge of the genome sequence of Novius is a major step towards further understanding its biology and potential applications in pest control. Results We report the first high-quality genome sequence for Novius pumilus (Weise, 1892), a representative specialist of Novius. Computational Analysis of gene Family Evolution (CAFE) analysis showed that several orthogroups encoding chemosensors, digestive, and immunity-related enzymes were significantly expanded (P

Species of the genus Oxyscelio Kieffer are egg parasitoids of Orthoptera. The genus is relatively diverse in China, with 34 described species. Some species of the genus are extremely morphologically similar and difficult to identify, especially in males. In this study, DNA barcoding based on sequences of the COI gene was used to discriminate Oxyscelio species from southern China. In total, 49 COI sequences belonging to 22 morphospecies were obtained. The COI sequences worked well for the identification of all the studied species, with intraspecific genetic distances ranging between 0 and 4.3%, while interspecific distances ranged between 7.1% and 19%. Based on both morphological and molecular analyses, five species are described as new: O. amalocarina Mo & Chen, sp. nov., O. apheles Mo & Chen, sp. nov., O. latheticus Mo & Chen, sp. nov., O. stenos Mo & Chen, sp. nov., and O. striae Mo & Chen, sp. nov.

The relictual archostematan beetle family Ommatidae attained high diversity during the Mesozoic. Despite their once high taxonomic diversity and morphological disparity, many Mesozoic ommatid taxa remain poorly understood, partly due to limited preservation. Here we report an exceptionally well-preserved fossil, which we describe as a new ommatid genus and species, Limnomma daohugouense gen. et sp. nov., from the mid-Jurassic Daohugou Lagerstätte in Northeast China. The new genus can be most easily distinguished from other ommatids by the presence of a circular non-tuberculate region on ventrite 5. The new taxon is discussed in relation to the classification of the Mesozoic genera Brochocoleus and Burmocoleus.

A new genus and species, Multistria orthotropa gen. et sp. nov., and a new species, Dachibangus hui sp. nov., of Mimarachnidae are described from the mid-Cretaceous Burmese amber. These new taxa display unique wing color patterns and extend the Mesozoic diversity of Mimarachnidae. The evolution of wing venation, phylogenetic placement of Mimarachnidae, and anti-predation defenses of this family in Burmese amber forest are briefly discussed.

2-dose measles-mumps-rubella (MMR) vaccine was recommended for children in Shanghai in November 1996 and incorporated into Shanghai immunization program in December 2008. We described the mumps epidemiology and assessed impact of the 2-dose MMR vaccination in Changning district, Shanghai, 1990–2017. We obtained the MMR vaccination coverage for children born during 1995–2015 and examined the incidence and disease characteristics of mumps during 1990–2017. The 1st dose MMR coverage had maintained above 95% since 1999 birth cohort. The 2nd dose MMR coverage reached above 90% since 2006 birth cohort. A total of 13,388 cases were reported during 1990–2017. The incidence decreased from 315.2 per 100,000 population in 1990 to 8.8 per 100,000 population in 2017. Of the 13,388 cases, 7585 (56.7%) were male and 91.7% were 1–14 years of age and 86.8% were children in kindergartens and students in schools. Compared with 1990–1996, the incidence had a significant decrease in 0–4 and ≥15 years in 1997–2008 and in all age groups in 2009–2017. A later birth cohort was associated with a lower incidence in children covered by MMR vaccination. In Conclusions, the incidence of mumps has dramatically declined with high coverage of 2-dose MMR in Changning district, Shanghai. Children in kindergartens and schools are still the most affected populations. An increase in incidence in adults has not occurred after 20 years of MMR vaccination. Long-term surveillance is needed to fully evaluate the impact of MMR vaccination policy.

Co-infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and other viruses has been reported. We evaluated cell lines commonly used to isolate viruses and diagnose related diseases for their susceptibility to SARS-CoV-2. Although multiple kidney cell lines from monkeys were susceptible to SARS-CoV-2, we found many cell types derived from humans, dogs, minks, cats, mice, and chicken were not. We analyzed MDCK cells, which are most commonly used for surveillance and study of influenza viruses, and found that they were not susceptible to SARS-CoV-2. The low expression level of the angiotensin converting enzyme 2 receptor and lower receptor affinity to SARS-CoV-2 spike, which could be overcome by overexpression of canine angiotensin converting enzyme 2 in trans, strengthened the cellular barrier to productive infection. Moreover, a D614G mutation in the spike protein did not appear to affect SARS-CoV-2 cell tropism. Our findings should help avert inadvertent propagation of SARS-CoV-2 from diagnostic cell lines.

Abstract Background The ladybird beetle Cryptolaemus montrouzieri Mulsant, 1853 (Coleoptera, Coccinellidae) is used worldwide as a biological control agent. It is a predator of various mealybug pests, but it also feeds on alternative prey and can be reared on artificial diets. Relatively little is known about the underlying genetic adaptations of its feeding habits. Results We report the first high-quality genome sequence for C. montrouzieri. We found that the gene families encoding chemosensors and digestive and detoxifying enzymes among others were significantly expanded or contracted in C. montrouzieri in comparison to published genomes of other beetles. Comparisons of diet-specific larval development, survival and transcriptome profiling demonstrated that differentially expressed genes on unnatural diets as compared to natural prey were enriched in pathways of nutrient metabolism, indicating that the lower performance on the tested diets was caused by nutritional deficiencies. Remarkably, the C. montrouzieri genome also showed a significant expansion in an immune effector gene family. Some of the immune effector genes were dramatically downregulated when larvae were fed unnatural diets. Conclusion We suggest that the evolution of genes related to chemosensing, digestion, and detoxification but also immunity might be associated with diet adaptation of an insect predator. These findings help explain why this predatory ladybird has become a successful biological control agent and will enable the optimization of its mass rearing and use in biological control programs.

Hydrocarbon potential evaluation is a high priority in oil/gas exploration which is significantly influenced by evaluation methods and subjective cognition. With the aim of quantitatively establishing a hydrocarbon generation and expulsion (HGE) model of source rocks, a data-driven approach is proposed based on abundant and readily available Rock-Eval/total organic carbon content (TOC)/vitrinite reflectance (VRo) datasets. This approach takes into account the original hydrocarbon generation potential (GPIo) and the loss of TOC. Hydrocarbon generation simulation was also adopted in this study. This data-driven method was applied to the Lower Paleogene Shahejie Formation (Es4), which has three sets of superior source rocks (average thickness > 250 m, TOC > 3%). The GPIo of the Es4 low-maturity source rock was 600 mg HC/g TOC. The initial maturity values of hydrocarbon generation and expulsion were 0.36% and 0.46%, respectively. The Gaosheng sub-member of the Es4 source rock had the largest amount of hydrocarbon generation but a limited amount of hydrocarbon expulsion, implying a favorable exploration of shale oil resources. By contrast, the values of the conventional, unconventional tight, and shale oil resource potentials of the Niuxintuo sub-member were 3.20 × 107, 7.70 × 107, and 5.93 × 108 t, respectively, indicating good prospects for tight oil and shale oil exploration. A comparison between the previous method and the data-driven method showed considerable differences in the restoration of GPIo and TOC, and both had their own limitations. This data-driven approach provides a quick and valid source rock evaluation method and can greatly enhance the accuracy of resource assessment.

Abstract Background Horizontal gene transfer (HGT) has been documented in many herbivorous insects, conferring the ability to digest plant material and promoting their remarkable ecological diversification. Previous reports suggest HGT of antibacterial enzymes may have contributed to the insect immune response and limit bacterial growth. Carnivorous insects also display many evolutionary successful lineages, but in contrast to the plant feeders, the potential role of HGTs has been less well-studied. Results Using genomic and transcriptomic data from 38 species of ladybird beetles, we identified a set of bacterial cell wall hydrolase (cwh) genes acquired by this group of beetles. Infection with Bacillus subtilis led to upregulated expression of these ladybird cwh genes, and their recombinantly produced proteins limited bacterial proliferation. Moreover, RNAi-mediated cwh knockdown led to downregulation of other antibacterial genes, indicating a role in antibacterial immune defense. cwh genes are rare in eukaryotes, but have been maintained in all tested Coccinellinae species, suggesting that this putative immune-related HGT event played a role in the evolution of this speciose subfamily of predominant predatory ladybirds. Conclusion Our work demonstrates that, in a manner analogous to HGT-facilitated plant feeding, enhanced immunity through HGT might have played a key role in the prey adaptation and niche expansion that promoted the diversification of carnivorous beetle lineages. We believe that this represents the first example of immune-related HGT in carnivorous insects with an association with a subsequent successful species radiation.

Eucinetus parvus sp. nov. is described from late Cretaceous Burmese amber, representing the second record of Eucinetidae from the Burmese amber and the first species of the family with simple, not piercing, mouthparts. A comparison between Mesocinetidae and Eucinetidae is provided.

Gonatopus viet Olmi, 1986 was originally described from Vietnam based on a single female. No further distribution records or hosts have been documented since its original discovery. In the present study, this species is newly recorded from China and its host is confirmed as Stirellus capitatus (Distant, 1918) using DNA barcoding techniques. The utility of DNA barcoding to discover host-Dryinidae associations is discussed.

Two new species of Trigonalyidae are described from Tibet (SW China): Jezonogonalos nyingchiensis Chen & van Achterberg, sp. nov., and Taeniogonalos eurysoma Chen & van Achterberg, sp. nov. In total, seven species representing four genera are known from Tibet, and two of them are newly recorded from Tibet: Taeniogonalos bucarinata Chen, van Achterberg, He & Xu, 2014, and Teranishia crenulata Chen, van Achterberg, He & Xu, 2014.

Three cryptic species, which were previously reported as Nidirana adenopleura, are revealed on the basis of comprehensive approaches. Nidirana guangdongensis Lyu, Wan, and YY Wang, sp. nov. is distributed in Nanling Mountains and southern Luoxiao Mountains, Nidirana mangveni Lyu, Qi, and YY Wang, sp. nov. is known from northern Zhejiang, and Nidirana xiangica Lyu and YY Wang, sp. nov. occurs in Xiangjiang River Basin, while the true Nidirana adenopleura is designated from Taiwan Island, northern Fujian, southern Zhejiang, and central Jiangxi. These three new species can be distinguished from all congeners by significant divergences in the mitochondrial 16S and CO1 genes, differences in advertisement calls, and the combination of multiple characteristics. This work indicates that the current records of Nidirana adenopleura should be of a species complex composed of multiple species and have clarified the true identity of N. adenopleura.

Abstract Background Ladybird beetles (Coleoptera, Coccinellidae) are highly diverse in their feeding habits. Most of them are specialist feeders, while some can have a broad spectrum of prey. As a representative group of generalists, the tribe Coccinellini includes many aphidophagous species, but members of this tribe also feed on other hemipterous insects including coccids, psyllids and whiteflies. As a result, several species are effective biological control agents or invasive species with serious non-target effects. Despite their economic importance, relatively little is known about how they adapt to new prey. Results In this study, comparisons of the life history traits and transcriptomes of ladybirds fed initial (aphids) and alternative prey (mealybugs) were performed in three Coccinellini species. The use of alternative prey greatly decreased performance, implied by the significantly prolonged development time and decreased survival rate and adult weight. Prey shifts resulted in a set of differentially expressed genes encoding chemosensory proteins and digestive and detoxifying enzymes. Conclusions Our results suggest that these generalists do not perform well when they use alternative prey as the sole nutrition source. Although their capacity for predation might have created an opportunity to use varied prey, they must adapt to physiological obstacles including chemosensing, digestion and detoxification in response to a prey shift. These findings challenge the effect of Coccinellini predators on the biological control of non-aphid pests and suggest the possibility of non-target attacks by so-called specialists.

The fine-grained rocks in the Paleogene Shahejie Formation in Nanpu Sag, Huanghua Depression, Bohai Bay Basin, are extremely important source rocks. These Paleogene rocks are mainly subdivided into organic-rich black shale and gray mudstone. The average total organic carbon contents of the shale and mudstone are 11.5 wt.% and 8.4 wt.%, respectively. The average hydrocarbon (HC)-generating potentials (which is equal to the sum of free hydrocarbons (S 1 ) and potential hydrocarbons (S 2 )) of the shale and mudstone are 39.3 mg HC/g rock and 28.5 mg HC/g rock, respectively, with mean vitrinite reflectance values of 0.82% and 0.81%, respectively. The higher abundance of organic matter in the shale than in the mudstone is due mainly to paleoenvironmental differences. The chemical index of alteration values and Na/Al ratios reveal a warm and humid climate during shale deposition and a cold and dry climate during mudstone deposition. The biologically derived Ba and Ba/Al ratios indicate high productivity in both the shale and mudstone, with relatively low productivity in the shale. The shale formed in fresh to brackish water, whereas the mudstone was deposited in fresh water, with the former having a higher salinity. Compared with the shale, the mudstone underwent higher detrital input, exhibiting higher Si/Al and Ti/Al ratios. Shale deposition was more dysoxic than mudstone deposition. The organic matter enrichment of the shale sediments was controlled mainly by reducing conditions followed by moderate-to-high productivity, which was promoted by a warm and humid climate and salinity stratification. The organic matter enrichment of the mudstone was less than that of the shale and was controlled by relatively oxic conditions.

In recent years, a large amount of industrial oil and gas resources have been discovered in the Paleogene Wenchang Formation (WC) of the southern Lufeng sag, confirming that the WC resources are promising prospects for petroleum exploration. However, because of the complex lithology, multiple sources of crude oil and multi-period charging characteristics in the WC, the accumulation process and model have not been clearly understood. Therefore, in this study, the main sources of crude oil and the process of hydrocarbon accumulation in key oil accumulation periods are determined by combining biomarker and geological analyses. Finally, the model of oil and gas accumulation is systematically summarized. The obtained results show that the source rock of the WC is the main source of hydrocarbons in the study area. The crude oil types in the southern Lufeng sag are classified into three types. Type A crude oil comes from source rocks of Wenchang Formation Members 3 and 4. Type B crude oil comes partly from source rocks of Wenchang Formation Members 1, 2, and 3 and partly from source rocks of Wenchang Formation Members 5 and 6. Type C crude oil is a mix of type A and type B crude oil. There are three accumulation periods that typify the study area: in the first period, accumulation was with vertical migration by fault; in the second period, accumulation was with lateral migration by sand body; and in the third period, accumulation was with vertical migration by sand body and adjacent to the source. The methodology and results of the accumulation process in southern Lufeng sag can support offshore oil and gas exploration.

ABSTRACT Laboratory surveillance for poliovirus (PV) relies on virus isolation by cell culture to identify PV in stool specimens from acute flaccid paralysis (AFP) cases. Although this method successfully identifies PV, it is time-consuming and necessitates the additional biorisk of growing live virus in an increasingly polio-free world. To reduce the risk of culturing PV, the Global Polio Laboratory Network (GPLN) must switch to culture-independent diagnostic methods with sensitivity at least equivalent to that of cell culture procedures. Five commercial nucleic acid extraction kits and one enrichment method were tested for PV extraction efficiency. RNA yield was measured using real-time reverse transcription (RT)-PCR. Based on greater RNA yield, compared with the other kits, the Quick-RNA viral kit was selected for further testing and was optimized using an RNA extraction procedure for stool suspensions. RNA extraction was retrospectively tested with 182 stool samples that had previously tested positive for PVs, in parallel with the standard GPLN virus isolation algorithm. After virus isolation or RNA extraction, real-time RT-PCR assays were performed. RNA extraction was significantly more sensitive than virus isolation (McNemar’s test, P < 0.001). Thereafter, the RNA extraction method was tested in parallel for 202 prospective samples; RNA extraction and virus isolation were not significantly different from each other (McNemar’s test, P = 0.13). Direct RNA extraction was noninferior to current cell culture methods for detecting PV in stool samples. Our results show that direct RNA extraction can make downstream manipulation safer and can reduce the risk of accidental posteradication viral release. The method is amenable to implementation in a wide variety of polio laboratories. IMPORTANCE Successfully identifying poliovirus from acute flaccid paralysis (AFP) cases is a vital role of the Global Polio Laboratory Network to achieve the goals of the Global Polio Eradication Initiative. Currently, laboratory surveillance relies on virus isolation by cell culture to test for PV present in stool samples. Although this method can identify polioviruses, laboratories must switch to culture-independent methods to reduce the risk associated with growing live viruses in a soon-to-be polio-free world. By implementing this streamlined method, in combination with real-time RT-PCR, laboratories can quickly screen for and type polioviruses of programmatic importance to support the final stages of global polio eradication.

A new species of Music frogs, Nidirana yaoica sp. nov. is described based on a series of adult male specimens collected from Mt Dayao, Guangxi, southern China, providing valuable new information on the phylogeny, bioacoustics, and biogeography of related species within the genus Nidirana. The new species forms the sister taxon to N. daunchina from western China and together the sister taxon to N. chapaensis from northern Vietnam. Nidirana yaoica sp. nov. can be distinguished from all known congeners by a significant genetic divergence in the mitochondrial 16S and CO1 genes, the advertisement call containing 1–3 rapidly repeated regular notes, and the combination of morphological characteristics including a medium-sized body with SVL 40.4–45.9 mm in adult males; lateroventral grooves on every digit, not meeting at the tip of disk; tibio-tarsal articulation reaching the nostril; the presence of a pair of subgular vocal sacs in males; and one single developed nuptial pad on dorsal surface of first finger in males.

Abstract Biological control is the main purpose of intentionally introducing non‐native invertebrate species. The evolutionary changes that occur in the populations of the introduced biological control agents may determine the agent's efficiency and the environmental safety. Here, to explore the pattern and extent of potential genomic changes in the worldwide introduced predatory ladybird beetle Cryptolaemus montrouzieri, we used a reduced‐representation sequencing method to analyze the genome‐wide differentiation of the samples from two native and five introduced locations. Our analyses based on a total of 53,032 single nucleotide polymorphism loci showed that beetles from the introduced locations in Asia and Europe exhibited significant reductions in genetic diversity and high differentiation compared with the samples from the native Australian range. Each introduced population belonged to a unique genetic cluster, while the beetles from two native locations were much more similar. These genomic patterns were also detected when the dataset was pruned for genomic outlier loci (52,318 SNPs remaining), suggesting that random genetic drift was the main force shaping the genetic diversity and population structure of this biological control agent. Our results provide a genome‐wide characterization of polymorphisms in a biological control agent and reveal genomic differences that were influenced by the introduction history. These differences might complicate assessments of the efficiency of biological control and the invasion potential of this species but also indicate the feasibility of selective breeding.

Alongiside the growing demand for wearable and implantable electronics, the development of flexible thermoelectric (FTE) materials holds great promise and has recently become a highly necessitated and efficient method for converting heat to electricity. Conductive polymers were widely used in previous research; however, n-type polymers suffer from instability compared to the p-type polymers, which results in a deficiency in the n-type TE leg for FTE devices. The development of the n-type FTE is still at a relatively early stage with limited applicable materials, insufficient conversion efficiency, and issues such as an undesirably high cost or toxic element consumption. In this work, as a prototype, a flexible n-type rare-earth free skutterudite (CoSb3)/poly(3,4-ethylenedioxythiophene)-polystyrene sulfonate (PEDOT:PSS) binary thermoelectric film was fabricated based on ball-milled skutterudite via a facile top-down method, which is promising to be widely applicable to the hybridization of conventional bulk TE materials. The polymers bridge the separated thermoelectric particles and provide a conducting pathway for carriers, leading to an enhancement in electrical conductivity and a competitive Seebeck coefficient. The current work proposes a rational design towards FTE devices and provides a perspective for the exploration of conventional thermoelectric materials for wearable electronics.

Polioviruses are positive-sense, single-stranded RNA picornaviruses and the principal cause of poliomyelitis. Global poliovirus surveillance has relied on poliovirus isolation in cells, which may take a minimum of 10 days, involves maintaining two cell lines, and propagates virus in high titers. With eradication underway, a major objective of the Global Polio Eradication Initiative (GPEI) is to develop culture-independent detection of polioviruses as an alternative method to complement the current virus isolation technique. A culture-independent method on poliovirus-positive stool suspensions was assessed with commercially available recombinant soluble poliovirus receptor (PVR) coupled to Histidine (His) tags. Viral RNA was screened by quantitative real-time reverse transcription PCR using the poliovirus intratypic differentiation kit. Poliovirus recovery was optimized with PVR-His–tagged protein and buffers supplemented with polyethylene glycol. To validate the poliovirus-PVR–His tag purification assay, 182 poliovirus-positive stools of programmatic importance were parallel tested against the GPLN-accepted virus isolation method. The PVR-His tag enrichment method detected poliovirus in 164 of 171 poliovirus-positive stools, whereas the virus isolation method misidentified 38 stools as poliovirus-negative (McNemar χ2p

Tight oil is an important unconventional resource, and sand production is an inevitable challenge during the field development. In this paper, based on data from the Daqing oilfield in Songliao Basin, the sand production of the tight sandstone oil reservoir is studied from the perspective of seepage and in situ stress distribution. Based on the combination of the formation fluid seepage law and the stress distribution around the well, a sand production prediction model is proposed to quantitatively estimate the sand production rate. The sand production prediction model is built based on the derivation of the sand production rate, which is well validated against the field data in the Daqing field with a relative error of 4.38%.The following conclusions are drawn: (1) after the critical pressure difference is exceeded, the sand production rate is smaller with a higher flowing bottom-hole pressure; (2) a smaller sand production radius makes the formation more unstable and causes a more severe sand production; and (3) various sand production rates exhibit due to different permeabilities. A larger permeability results in a higher sand production rate. The findings of this study can help for sand production prediction in the tight sandstone oil reservoir.

Abstract Background Introduced biological control agents have opportunities of population admixture through multiple introductions in the field. However, the importance of population admixture for their establishment success often remains unclear. Previous studies based on genetic markers have suggested a history of population admixture in the predatory ladybird Cryptolaemus montrouzieri Mulsant in China. Results We tested whether population admixture may lead to fitness changes under laboratory conditions. We first found no mating barrier or strong bias between two parental populations, despite their differences in genetics and phenotypes. Then, our experimental evidence supported the hypothesis that admixed populations have a higher potential of establishment success, due to their superior reproductive ability, and hunger and cold tolerance inherited from one of the parental populations. Conclusions We suggest that population admixture can be a breeding method to improve the performance of biological control agents, particularly when used in a classical biological control approach, but that consequences for potential invasiveness need to be considered.

A new genus and species of Sisyridae, Stictosisyra pennyi gen. et sp. n., is described from the Late Cretaceous (earliest Cenomanian/late Albian) Myanmar amber. It can be easily distinguished from other sisyrids genera by the configuration of wing venation such as forewing with four ra-rp crossveins, M forked distal to the separation of RP1, CuA pectinate and CuP simple; hind wing 1r-m long and sinuous. Besides, the newly documented spongillaflies bore distinct, irregularly distributed spots on the forewings.

The phylogeny of beetles, which represent ~25% of known extant animal species, has been a challenge to resolve. Here, Zhang et al. infer a time-calibrated phylogeny for Coleoptera based on 95 protein-coding genes in 373 species and suggest an association between the hyperdiversification of beetles and the rise of angiosperms.

A new species of Hemerobiidae, Wesmaelius makarkini Yang, Pang & Ren, sp. n. is described from the Lower Miocene, Garang Formation of Zeku County, Qinghai Province (northeastern Tibetan Plateau), China. The species is assigned to the widely distributed extant genus Wesmaelius Krüger (Hemerobiinae). The species represents the first named fossil of this family from China, which sheds light on the historical distribution of Wesmaelius and early divergences within Hemerobiinae.

A new genus and species of the cleroid family Lophocateridae are described and illustrated from the mid-Cretaceous amber of northern Myanmar. Gracilenticrus burmiticus Yu, Kolibáč & Ślipiński gen. et sp. nov. is unique among Lophocateridae in the tiny body size, frontoclypeal suture and antennal grooves absent, symmetrical antennal clubs, protrochantin reduced, tarsal claws small and widened at base. A key to the species of Mesozoic Lophocateridae is also provided. Morphological characters of the newly discovered Gracilenticrus were analyzed together with representatives of 43 extant genera of Cleroidea (broadly defined Trogossitidae) in a matrix of 91 characters. Gracilenticrus burmiticus was resolved as a member of Lophocateridae. The discovery of a diverse fauna of Lophocateridae in the mid-Cretaceous sheds a new light on the early evolution of superfamily Cleroidea.

Eucalyptus species have become one of the most commonly planted trees worldwide, including China, due to their fast growth and various commercial applications. However, the productivity of Eucalyptus plantations has been threatened by exotic invasive insect pests in recent years. Among these pests, gall inducers of the genus Ophelimus of the Eulophidae family are among the most important invasive species in Eucalyptus plantations. We report here for the first time the presence of a new invasive Eucalyptus gall wasp, Ophelimus bipolaris sp. n., in Guangzhou, China, which also represents the first species of the genus reported from China. The identity of the new species was confirmed by an integrative approach combing biological, morphological and molecular evidence. The new species is described and illustrated. This wasp induces galls only on the leaf blade surface of four Eucalyptus species: E. grandis, E. grandis × E. urophylla, E. tereticornis and E. urophylla. Our preliminary observation showed that O. bipolaris could complete a life cycle on E. urophylla in approximately 2 months under local climatic conditions (23.5–30 °C). Considering the severe damage it may cause to Eucalyptus production, further investigations of its biology and control are urgently needed in China.

Abstract Background The tribe Coccinellini is a group of relatively large ladybird beetles that exhibits remarkable morphological and biological diversity. Many species are aphidophagous, feeding as larvae and adults on aphids, but some species also feed on other hemipterous insects (i.e., heteropterans, psyllids, whiteflies), beetle and moth larvae, pollen, fungal spores, and even plant tissue. Several species are biological control agents or widespread invasive species (e.g., Harmonia axyridis (Pallas)). Despite the ecological importance of this tribe, relatively little is known about the phylogenetic relationships within it. The generic concepts within the tribe Coccinellini are unstable and do not reflect a natural classification, being largely based on regional revisions. This impedes the phylogenetic study of important traits of Coccinellidae at a global scale (e.g. the evolution of food preferences and biogeography). Results We present the most comprehensive phylogenetic analysis of Coccinellini to date, based on three nuclear and one mitochondrial gene sequences of 38 taxa, which represent all major Coccinellini lineages. The phylogenetic reconstruction supports the monophyly of Coccinellini and its sister group relationship to Chilocorini. Within Coccinellini, three major clades were recovered that do not correspond to any previously recognised divisions, questioning the traditional differentiation between Halyziini, Discotomini, Tytthaspidini, and Singhikaliini. Ancestral state reconstructions of food preferences and morphological characters support the idea of aphidophagy being the ancestral state in Coccinellini. This indicates a transition from putative obligate scale feeders, as seen in the closely related Chilocorini, to more agile general predators. Conclusions Our results suggest that the classification of Coccinellini has been misled by convergence in morphological traits. The evolutionary history of Coccinellini has been very dynamic in respect to changes in host preferences, involving multiple independent host switches from different insect orders to fungal spores and plants tissues. General predation on ephemeral aphids might have created an opportunity to easily adapt to mixed or specialised diets (e.g. obligate mycophagy, herbivory, predation on various hemipteroids or larvae of leaf beetles (Chrysomelidae)). The generally long-lived adults of Coccinellini can consume pollen and floral nectars, thereby surviving periods of low prey frequency. This capacity might have played a central role in the diversification history of Coccinellini.

Two new species the genus Rhorus Förster are described from Thailand, R. inthanonensis Reshchikov & Xu, sp. n. and R. lannae Reshchikov & Xu, sp. n. from Chiang Mai Province. These are the first records of the genus from Thailand. The type specimen of R. orientalis (Cameron, 1909) is re-described and illustrated. An identification key for the Oriental species of Rhorus is provided.

A new species of the genus Lethades Davis, 1897 (Ctenopelmatinae: Pionini), L. orientalis Reshchikov & Xu, sp. n., collected in Heishiding Nature Reserve in Guangdong Province, China, is described. This is new record of the genus from China and for the Oriental region also. The species can be distinguished from all other species of the genus by its black metasoma, the presence of an areolet on the fore wing, distinctly pectinate claws with teeth of the hind claw as high as the claw itself, and a complete longitudinal propodeal carina. A key to the world species of the genus is provided.

The cotton mealybug Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae), is an emerging invasive insect pest in China. Hymenopteran parasitoids are the key organisms for suppressing populations of P. solenopsis in the field, and therefore could be used as biological agents. Accurate identification of the associated parasitoids is the critical step to assess their potential role in biological control. In this study, we facilitated the identification of the parasitoid composition of P. solenopsis using an integrated approach of species delimitation, combining morphology with molecular data. Eighteen Hymenoptera parasitoid species belonging to 11 genera of four families are recognized based on morphological examination and molecular species delimitation of the mitochondrial cytochrome c oxidase 1 (COI) gene and the 28S rDNA using the automatic barcode gap discovery (ABGD) and the Bayesian Poisson tree processes model (bPTP). Among these species, eight species are primary parasitoids with Aenasius arizonensis (Girault) (Hymenoptera: Encyrtidae) being the dominant taxon, while the other 10 species are probably hyperparasitoids, with a prevalence of Cheiloneurus nankingensis Li & Xu (Hymenoptera: Encyrtidae). These results indicate that parasitoid wasps associated with P. solenopsis from China are diverse and the integrated taxonomic approach applied in this study could enhance the accurate identification of these parasitoids that should be assessed in future biological control programs.

Enrofloxacin is a poorly soluble antibacterial drug of the fluoroquinolones class used in veterinary medicine. The main purpose of this work was to investigate the structural and pharmaceutical properties of new enrofloxacin salts. Enrofloxacin anhydrate and its organic salts with tartaric acid, nicotinic acid and suberic acid formed as pure crystalline anhydrous solids. All the crystals were grown from a mixed solution by slow evaporation at room temperature. These products were then characterized by field-emission scanning electron microscopy, powder X-ray diffraction, Fourier transform infrared spectroscopy and differential scanning calorimetry. Further, X-ray single crystal diffraction analysis was used to study the crystal structure. The intermolecular interactions and packing arrangements in the crystal structures were studied, and the solubility of these salts in water was determined using high-performance liquid chromatography. The results show that the new salts of enrofloxacin developed in this study exhibited excellent water solubility.

Plenty of marine crude oil generated by the Middle–Upper Ordovician source rocks has been discovered in the Tabei Uplift, Tarim Basin. Previous studies on the extensively distributed low organic matter source rock total organic carbon ≤ 0.5%) imply that this contributes to oil accumulation. Thus, the identification of present-day low-total organic carbon source rocks that experience hydrocarbon expulsion and evaluation of their contribution is of considerable significance to analyze marine oil accumulation and to assess the resources in the Tarim Basin. Using the mass balance approach, we identify and verify present-day low-total organic carbon carbonate source rocks and present a quantitative model using well-logging data to predict the present-day low-total organic carbon carbonate source rocks in order to evaluate their contribution in the Tarim Basin. Hydrocarbon expulsion has occurred in the Yijianfang formation, which has present-day low-total organic carbon, between 6452 and 6487 m in the Yangwu 2 well. In the horizons depicting carbonates with hydrocarbon expulsion, the porosity is relatively high and the daily oil production is 0.19 m 3 according to the perforation tests. The extracts have similar biomarker signatures to that of the crude oils in the Tabei Uplift. In this study, a quick and cost-effective quantitative model is developed using double lateral resistivity and uranium/thorium well-logging data. By applying the prediction model, the contribution of present-day low-total organic carbon carbonate source rocks in the Middle–Upper Ordovician formation of the Tarim Basin is primarily evaluated to be 363.38 × 10 9 bbl oil equivalent.

Biological control agents usually suffer from a shortage of target prey or hosts in their post-release stage. Some predatory agents turn to attacking other prey organisms, which may induce physiological and evolutionary changes. In this study, we investigated life history traits, gene expression and genotype frequency in the predatory ladybird beetle Cryptolaemus montrouzieri during experimental prey shifts. C. montrouzieri were either continuously fed on aphids Megoura japonica as an alternative prey for four generations or were shifted back to the initial prey mealybugs Planococcus citri in each generation. In general, the utilization of aphids resulted in reduced performance and severe physiological adjustments, indicated by significant changes in development and fecundity traits and a large number of differentially expressed genes between the two offering setup prey treatments. Within the aphid-fed lines, performance regarding the developmental time, the adult weight and the survival rate recovered to some level in subsequent generations, possibly as a result of adaptive evolution. In particular, we found that a shift back to mealybugs caused a gradual increase in fecundity. Accordingly, a genotype of the fecundity-related gene vitellogenin, of which there were several minor alleles in the initial population, became the main genotype within four generations. The present study explored the short-term experimental evolution of a so-call specialist predator under prey shift conditions. This potential rapid adaptation of biological control agents to novel prey will increase environmental risks associated with non-target effects.

A new species of Ripiphoridae Gemminger & Harold, 1870, Archaeoripiphorus nuwa gen. et sp. nov., is described and illustrated from a well-preserved impression fossil from the Middle Jurassic Jiulongshan Formation collected at Daohugou Village, Shantou Township, Ningcheng County, Inner Mongolia, China, representing the oldest documented occurrence of the Ripiphoridae described from the Mesozoic era. It shares several characters belonging to two basal ripiphorid subfamilies (Pelecotominae and Ptilophorinae), but it cannot be attributed to either of them and is herein placed as Subfamily incertae sedis. An overall similarity between Archaeoripiphorus gen. nov. and Recent Pelecotominae and the occurrence of wood-boring beetles in the same Formation implies a similar parasitoid host preference in xylophagous beetles for A. nuwa gen. et sp. nov., putting a spotlight on a potential host-parasitoid relationship in the Mesozoic.

This paper presents an integrated workflow for investigating hydrocarbon charge history using fluid inclusions using the Ordovician reservoirs from the Tazhong Oilfield, Tarim Basin as an example. The work flow involves the delineation of fluid inclusion assemblage (FIA) using fluid inclusion petrography and spectroscopy for microthermometric analysis. The interpretation of microthermometric data is based on data derived from synthetic inclusion experiments and takes consideration of the P-T re-equilibration of fluid inclusions in carbonate and over pressure effect on fluid inclusion trapping. The workflow also emphasizes on the requirements of adequate sample preparation, FIA identification, quantitative spectroscopic data (CIE), coeval petroleum and aqueous fluid inclusions, adequate number of microthermometric measurements and integrated interpretation using borehole temperature. In the study reservoirs in the Tazhong area, fluid inclusion spectroscopy reveals the presence of three groups of oil inclusions of near yellow, near blue and near white fluorescence colours. The fluid inclusion microthermometry data indicate the presence of at least two predominant hydrocarbon fluid inclusion assemblages including: homogenous liquid phase yellow fluorescence inclusions and liquid phase oil and condensate fluid inclusion assemblages with homogenous near blue, near white and near yellow fluorescence colours. The aqueous inclusions from the Ordovician reservoirs in the Tazhong area reveal the presence of low, moderate, high and very high salinities in the reservoirs. The extremely high salinity is believed to be caused by regional brine intrusions relating to regional structural movement and deformation. Key words: Tazhong Ordovician reservoirs, fluid inclusions, analytical procedure, fluid inclusion assemblages, hydrocarbon charge history

Sinoeuthemis daohugouensis gen. et sp. n. is the first record of the isophlebiopteran family Euthemistidae from Middle Jurassic of northeast China, while previously this family was restricted to the early Late Jurassic Kazakhstan. This new finding allows us to emend the family diagnosis with hindwing characters. This new species shows a mixture of characters alternatively present in different genera of the two families Euthemistidae and Sphenophlebiidae.

Recently, several invasive mealybugs (Hemiptera: Pseudococcidae) have rapidly spread to Asia and have become a serious threat to the production of cotton including transgenic cotton. Thus far, studies have mainly focused on the effects of mealybugs on non-transgenic cotton, without fully considering their effects on transgenic cotton and trophic interactions. Therefore, investigating the potential effects of mealybugs on transgenic cotton and their key natural enemies is vitally important. A first study on the effects of transgenic cotton on a non-target mealybug, Ferrisia virgata (Cockerell) (Hemiptera: Pseudococcidae) was performed by comparing its development, survival and body weight on transgenic cotton leaves expressing Cry1Ac (Bt toxin) + CpTI (Cowpea Trypsin Inhibitor) with those on its near-isogenic non-transgenic line. Furthermore, the development, survival, body weight, fecundity, adult longevity and feeding preference of the mealybug predator Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) was assessed when fed F. virgata maintained on transgenic cotton. In order to investigate potential transfer of Cry1Ac and CpTI proteins via the food chain, protein levels in cotton leaves, mealybugs and ladybirds were quantified. Experimental results showed that F. virgata could infest this bivalent transgenic cotton. No significant differences were observed in the physiological parameters of the predator C. montrouzieri offered F. virgata reared on transgenic cotton or its near-isogenic line. Cry1Ac and CpTI proteins were detected in transgenic cotton leaves, but no detectable levels of both proteins were present in the mealybug or its predator when reared on transgenic cotton leaves. Our bioassays indicated that transgenic cotton poses a negligible risk to the predatory coccinellid C. montrouzieri via its prey, the mealybug F. virgata.

High quality single crystalline GaN nanowires with large aspect ratio (>100) are synthesized on n-type Si (111) substrate via Au-catalyzed vapor-liquid-solid process. Morphology, crystal structure, and optical property of the as-synthesized GaN nanowires are characterized by means of X-ray diffraction, scanning/transmission electron microscopy, UV-vis diffuse reflection spectroscopy, and room temperature photoluminescence. The results indicate that the as-prepared GaN nanowires with a large aspect ratio are well crystallized in the hexagonal wurtzite structure, and a slight blue shift appears in both the absorption edge and emission peak probably due to the quantization effect. Photocatalytic H2 evolution over the as-prepared GaN nanowires is performed with the incorporation of Pt or Rh as the cocatalyst, exhibiting greatly enhanced capability compared to the GaN powder tested under the same conditions. Moreover, photocatalytic CO2 reduction over the GaN nanowires is also successfully realized using Pt or Rh as the cocatalyst, depending on which the products show a strong selectivity inherently related to the reductive electrons transferred by cocatalyst.

BackgroundMating behaviors have been widely studied for extant insects. However, cases of mating individuals are particularly rare in the fossil record of insects, and most of them involved preservation in amber while only in rare cases found in compression fossils. This considerably limits our knowledge of mating position and genitalia orientation during the Mesozoic, and hinders our understanding of the evolution of mating behaviors in this major component of modern ecosystems.Principal findingHere we report a pair of copulating froghoppers, Anthoscytina perpetua sp. nov., referable to the Procercopidae, from the Middle Jurassic of northeastern China. They exhibit belly-to-belly mating position as preserved, with male's aedeagus inserting into the female's bursa copulatrix. Abdominal segments 8 to 9 of male are disarticulated suggesting these segments were twisted and flexed during mating. Due to potential taphonomic effect, we cannot rule out that they might have taken side-by-side position, as in extant froghoppers. Genitalia of male and female, based on paratypes, show symmetric structures.Conclusions/significanceOur findings, consistent with those of extant froghoppers, indicate froghoppers' genitalic symmetry and mating position have remained static for over 165 million years.