Hepialus Research Articles

A near chromosome-level genome assembly of a ghost moth (Lepidoptera, Hepialidae)

Ghost moths are an unusual family of primitive moths (Lepidoptera: Hepialidae) known for their large body size and crepuscular adult activity. These moths represent an ancient lineage, frequently have soil dwelling larvae, and are adapted to high elevations, deserts, and other extreme environments. Despite being rather speciose with more than 700 species, there is a dearth of genomic resources for the family. Here, we present the first high quality, publicly available hepialid genome, generated from an Andean species of ghost moth, Druceiella hillmani. Our genome assembly has a length of 2,586 Mbp with contig N50 of 28.1 Mb and N50 of 29, and BUSCO completeness of 97.1%, making it one of the largest genomes in the order Lepidoptera. Our assembly is a vital resource for future research on ghost moth genomics.

Neurotransmitter acetylcholine mediates the mummification of Ophiocordyceps sinensis-infected Thitarodes xiaojinensis larvae.

Parasites can manipulate host behavior to facilitate parasite transmission. One such host-pathogen interaction occurs between the fungus Ophiocordyceps sinensis and the ghost moth Thitarodes xiaojinensis. O. sinensis is involved in the mummification process of infected host larvae. However, the underlying molecular and chemical mechanism for this phenomenon is unknown. We characterized the small molecules regulating host behaviors and the altered metabolites in infected and mummified host larvae. Lipid-related metabolites, such as phosphatidylcholine, were identified in infected and mummified larvae. Decreased levels of the neurotransmitter acetylcholine (ACh) and elevated choline levels were observed in the brains of both the infected and mummified larvae. The aberrant activity of acetylcholinesterase (AChE) and relative mRNA expression of ACE2 (acetylcholinesterase) may mediate the altered transformation between ACh and choline, leading to the brain dysfunction of mummified larvae. Caspofungin treatment inhibited the mummification of infected larvae and the activity of AChE. These findings indicate the importance of ACh in the mummification of host larvae after O. sinensis infection.IMPORTANCEOphiocordyceps sinensis-infected ghost moth larvae are manipulated to move to the soil surface with their heads up in death. A fruiting body then grows from the caterpillar's head, eventually producing conidia for dispersal. However, the underlying molecular and chemical mechanism has not been characterized. In this study, we describe the metabolic profile of Thitarodes xiaojinensis host larvae after O. sinensis infection. Altered metabolites, particularly lipid-related metabolites, were identified in infected and mummified larvae, suggesting that lipids are important in O. sinensis-mediated behavioral manipulation of host larvae. Decreased levels of the neurotransmitter acetylcholine were observed in both infected and mummified larvae brains. This suggests that altered or reduced acetylcholine can mediate brain dysfunction and lead to aberrant behavior. These results reveal the critical role of acetylcholine in the mummification process of infected host larvae.

Harposporium incensis sp. nov. a South American cordycipitoid species exhibiting inter-phylum host-jumping and having potential as a biological control agent for pest management

ABSTRACT Macro- and microscopic morphological studies and multilocus phylogenetic analysis were made on larval specimens of a ghost moth collected from a pigeon pea plantation in Huánuco, Peru. DNA sequences from the cadaver and the fungal isolates obtained represented a monophyletic clade based on the phylogeny. All morphological characters and molecular data showed that the pathogenic fungus infecting the ghost moth larvae was an unknown cordycipitoid species, herein described as, Harposporium incensis sp. nov. based on morphological features and multilocus phylogenetic analysis on the cadaver and fungus isolated from the same specimen. The far-related and ecologically different hosts of teleomorph and anamorph of this new species display a peculiar inter-phylum host jumping between the insect Trichophassus giganteus of the phylum Arthropoda and the nematode Caenorhabditis elegans of the phylum Nematoda and have biological control potential.

Tandemly expanded OR17b in Himalaya ghost moth facilitates larval food allocation via olfactory reception of plant-derived tricosane

Herbivorous insects utilize intricate olfactory mechanisms to locate food plants. The chemical communication of insect-plant in primitive lineage offers insights into evolutionary milestones of divergent olfactory modalities. Here, we focus on a system endemic to the Qinghai-Tibetan Plateau to unravel the chemical and molecular basis of food preference in ancestral Lepidoptera. We conducted volatile profiling, neural electrophysiology, and chemotaxis assays with a panel of host plant organs to identify attractants for Himalaya ghost moth Thitarodes xiaojinensis larvae, the primitive host of medicinal Ophiocordyceps sinensis fungus. Using a DREAM approach based on odorant induced transcriptomes and subsequent deorphanization tests, we elucidated the odorant receptors responsible for coding bioactive volatiles. Contrary to allocation signals in most plant-feeding insects, T. xiaojinensis larvae utilize tricosane from the bulbil as the main attractant for locating native host plant. We deorphanized a TxiaOR17b, an indispensable odorant receptor resulting from tandem duplication of OR17, for transducing olfactory signals in response to tricosane. The discovery of this ligand-receptor pair suggests a survival strategy based on food location via olfaction in ancestral Lepidoptera, which synchronizes both plant asexual reproduction and peak hatch periods of insect larvae.

A ghost moth olfactory prototype of the lepidopteran sex communication.

Sex role differentiation is a widespread phenomenon. Sex pheromones are often associated with sex roles and convey sex-specific information. In Lepidoptera, females release sex pheromones to attract males, which evolve sophisticated olfactory structures to relay pheromone signals. However, in some primitive moths, sex role differentiation becomes diverged. Here, we introduce the chromosome-level genome assembly from ancestral Himalaya ghost moths, revealing a unique olfactory evolution pattern and sex role parity among Lepidoptera. These olfactory structures of the ghost moths are characterized by a dense population of trichoid sensilla, both larger male and female antennal entry parts of brains, compared to the evolutionary later Lepidoptera. Furthermore, a unique tandem of 34 odorant receptor 19 homologs in Thitarodes xiaojinensis (TxiaOr19) has been identified, which presents overlapped motifs with pheromone receptors (PRs). Interestingly, the expanded TxiaOr19 was predicted to have unconventional tuning patterns compared to canonical PRs, with nonsexual dimorphic olfactory neuropils discovered, which contributes to the observed equal sex roles in Thitarodes adults. Additionally, transposable element activity bursts have provided traceable loci landscapes where parallel diversifications occurred between TxiaOr19 and PRs, indicating that the Or19 homolog expansions were diversified to PRs during evolution and thus established the classic sex roles in higher moths. This study elucidates an olfactory prototype of intermediate sex communication from Himalaya ghost moths.

Knockdown of Thitarodes host genes influences dimorphic transition of Ophiocordyceps sinensis in the host hemolymph.

The Chinese cordyceps, a unique parasitic complex of Thitarodes/Hepialus ghost moths and Ophiocordyceps sinensis fungus in the Tibetan Plateau, is a highly valuable biological resource for medicine and health foods in Asian countries. Efficient system for artificial cultivation of Chinese cordyceps relies on understanding the gene functions involved in the induction of growing blastospores into hyphae in the larval hemolymph of insect host, during O. sinensis infection. Transcriptome analysis and ribonucleic acid interference (RNA interference) method were employed to identify the key differentially expressed genes and to demonstrate their functions in Thitarodes xiaojinensis. Key larval genes critical for O. sinensis blastospore development or filamentation were identified. Nine of the 20 top upregulated genes encoded cuticles proteins, indicating that these proteins highly activated when the larval hemolymph was full of blastospores. Small interfering RNA (siRNA) knockdown of five larval genes such as Flightin, larval cuticle protein LCP-30, 26-hydroxylase (CYP18A1), cuticle protein 18.6, isoform B, and probable chitinase 3 significantly stimulated the dimorphic transition from blastospores to prehyphae in O. sinensis in the larval hemolymph after 120 h after injection. The expressions of these genes determined by quantitative real-time PCR were suppressed in various levels from 38.64% to 91.54%, compared to the controls. These results demonstrated that injection of the siRNAs of key upregulated genes into the larval hemolymph containing high load of blastospores caused the gene silence in T. xiaojinensis larvae and induced the fungal transition from blastospores to prehyphae, providing novel knowledge on the regulation of O. sinensis fungal dimorphism by Thitarodes host and cues for further study of Thitarodes biology and commercial cultivation of Chinese cordyceps.

EXPLORING THE MEDICINAL MARVEL: THE POTENTIAL OFYARSAGUMBA AS REGENERATIVE MEDICINE AND IN WOUND HEALING

Yarsagumba(Cordyceps mushroom) a valued herbal medicine, is a fungus that grows on ghost moth larvae. It belongs to the Ophiocordycipitaceae family and has been used in traditional Chinese medicine for centuries. It has various applications in the pharmaceutical and health industries. In Nepal, Yarsagumba has been recognized for thousands of years and is considered one of the most promising medicinal mushrooms. Scientists have been studying its unique life cycle and medical uses for the past 30 years.However, once correctly identified, these enigmatic organisms hold great promise in the prevention and treatment of numerous ailments such as cancer,woundhealing, antidiabetic, alleviating fatigue, respiratoryproblems, and erectile dysfunction in men.Cordyceps fungi possess significant phytochemicals, namely cordycepin and adenosine, which function by suppressing the adenosine A2 receptor and consequently inhibiting various neuroinflammatory markers such as NF-κB, NLRP3 inflammasome, IL-1β, iNOS, COX-2, TNF-α, and HMGB1. Yarsagumbas wound healing properties are multifaceted, with its anti-inflammatory and antioxidant compounds reducing inflammation and preventing tissue damage. Its immunomodulatory effects enhance the bodys immune response and inhibit the growth of harmful microorganisms, reducing the risk of infection. Yarsagumbas regenerative capabilities stimulate collagen production and promote angiogenesis, resulting in improved wound healing outcomes and minimized scarring. This review examines how Yarsagumbas bioactive compounds can enhance wound healing, reduce scarring, and improve tissue repair. It also explores the plants regenerative properties, such as its ability to stimulate stem cell growth and differentiation. Our findings indicate that the activation of adenosine receptors by adenosine and cordycepin effectively triggers the Wnt/β-catenin signaling, which can potentially facilitate tissue remodeling and serve as a therapeutic approach for skin wound treatment.

Plant quercetin degradation by gut bacterium Raoultella terrigena of ghost moth Thitarodes xiaojinensis.

Associated microbes of several herbivorous insects can improve insect fitness. However, the contribution of specific insect gut bacterium to plant toxin toxification for its host fitness remains scarce. Here, a gut bacterium Raoultella terrigena from the ghost moth Thitarodes xiaojinensis larvae was identified. This bacterium grew unhindered in the presence of Polygonum viviparum, which is a natural food for ghost moth larvae but showed significant growth inhibition and toxicity against Spodoptera litura. S. litura reared on artificial diets containing 5, 15 and 25% P. viviparum powder after 7 days coculture with R. terrigena were found to have shorter larval and pupal durations than on the diets containing P. viviparum powder but without R. terrigena coculture. HPLC analysis revealed that the content of quercetin in mineral medium containing 15% P. viviparum powder after 7 days coculture with R. terrigena was significantly decreased (79.48%) as compared with that in P. viviparum powder without R. terrigena coculture. In vitro fermentation further verified that R. terrigena could degrade 85.56% quercetin in Lucia-Bertani medium. S. litura reared on artificial diets containing 0.01, 0.05 and 0.1 mg/g quercetin after 48 h coculture with R. terrigena were also found to have shorter larval, prepupal and pupal durations, as well as higher average pupal weight and adult emergence rate than on the diets containing quercetin, but without R. terrigena coculture. In addition, R. terrigena was detected in the bud and root tissues of the sterilized P. viviparum, indicating that T. xiaojinensis larvae might acquire this bacterium through feeding. These results demonstrate that the gut bacteria contribute to the degradation of plant toxic molecules to improve the development of herbivorous insects and provide fundamental knowledge for developing effective methods for beneficial insect rearing and pest control.

Identification based on morphological and molecular characters of Phassus huebneri (Geyer) (Lepidoptera: Hepialidae), a ghost moth feeding on blackberry in Michoacán, Mexico: its biology and parasitoids

Larvae of an unidentified Lepidopteran hepialid were found boring stems and crown roots of blackberry (Rubus sp.) in Michoacán, Mexico. In this study, the morphological and molecular identification of larvae and adults of this insect was performed. Preliminary aspects of biology, including information on parasitoids associated to larval stage, are also recorded. A total of 109 larvae of various instars were collected and three were reared to adult. The two females and one male were identified as Phassus huebneri (Geyer) (Lepidoptera: Hepialidae) following morphological characters. This identification was corroborated by comparing the cytochrome oxidase I (COI) barcode of two field-collected larvae (IIAFL1 and IIAFL2) with sequences of Phassus spp. obtained from GenBank. A Neighbor-Joining similarity analysis yielded a phenogram with two subclades. The first subclade grouped the IIAFL1 and IIAFL2 sequences with two other Mexican sequences categorized as P. huebneri, whereas in the second subclade other species belonging to the genus Phassus from Guatemala and Costa Rica were grouped together. Larval development of nine instars took about 14 months. Larvae reached a maximum length of 10 cm. The pupal stage lasted 39-41 days. Each of the two females laid over 1,000 unfertilized eggs within a period of 48 h. Larvae were parasitized by Dinera grisescens Fallen (Diptera: Tachinidae) and another unidentified Dinera sp. This study provides additional evidences on the distribution pattern of P. huebneri in western Mexico and presents the first record of this species feeding on one of the most economically important crops in Mexico.

Transcriptomic Analysis Insight into the Immune Modulation during the Interaction of Ophiocordyceps sinensis and Hepialus xiaojinensis.

Ophiocordyceps sinensis (Berk.) is an entomopathogenic fungus that can infect the larva of the ghost moth, Hepialus xiaojinensis, causing mummification after more than one year. This prolonged infection provides a valuable model for studying the immunological interplay between an insect host and a pathogenic fungus. A comparative transcriptome analysis of pre-infection (L) and one-year post-infection (IL) larvae was performed to investigate the immune response in the host. Here, a total of 59,668 unigenes were obtained using Illumina Sequencing in IL and L. Among the 345 identified immune-related genes, 83 out of 86 immune-related differentially expressed genes (DEGs) had a much higher expression in IL than in L. Furthermore, the immune-related DEGs were classified as pathogen recognition receptors (PRRs), signal modulators or transductors, and immune effector molecules. Serpins and protease inhibitors were found to be upregulated in the late phase of infection, suppressing the host’s immune response. Based on the above analysis, the expression levels of most immune-related genes would return to the baseline with the immune response being repressed in the late phase of infection, leading to the fungal immunological tolerance after prolonged infection. Meanwhile, the transcriptomes of IL and the mummified larva (ML) were compared to explore O. sinensis invasion. A total of 1408 novel genes were identified, with 162 of them annotated with putative functions. The gene families likely implicated in O. sinensis pathogenicity have been identified, primarily including serine carboxypeptidase, peroxidase, metalloprotease peptidase, aminopeptidases, cytochrome P450, and oxidoreductase. Furthermore, quantitative real-time PCR (qPCR) was used to assess the expression levels of some critical genes that were involved in immune response and fungal pathogenicity. The results showed that their expression levels were consistent with the transcriptomes. Taken together, our findings offered a comprehensive and precise transcriptome study to understand the immune defense in H. xiaojinensis and O. sinensis invasion, which would accelerate the large-scale artificial cultivation of this medicinal fungus.

The invasion process of the entomopathogenic fungus Ophiocordyceps sinensis into the larvae of ghost moths (Thitarodes xiaojinensis) using a GFP-labeled strain.

Chinese cordyceps is a well-known and valuable traditional Chinese medicine that forms after Ophiocordyceps sinensis parasitizes ghost moth larvae. The low natural infection rate of O. sinensis limits large-scale artificial cultivation of Chinese cordyceps, and the invasion process is unclear. To investigate the temporal and spatial regulation when O. sinensis enters ghost moths, we constructed an O. sinensis transformant that stably expresses green fluorescent protein (GFP). Inoculating Thitarodes xiaojinensis larvae with a high concentration of GFP-labeled O. sinensis, we observed that O. sinensis conidia could adhere to the host cuticle within 2 days, germinate penetration pegs within 4 days, and produce blastospores in the host hemocoel within 6 days. The reconstructed three-dimensional (3D) structures of the invasion sites showed that penetration pegs germinated directly from O. sinensis conidia at the joining site with the larval cuticle. Differentiated appressoria or hyphae along the host epicuticle are not required for O. sinensis to invade ghost moths. Overall, the specific invasion process of O. sinensis into its host is clarified, and we provided a new perspective on the invasion process of entomopathogenic fungi.

New species of Phassus Walker ghost moth from northeastern Mexico and notes on its biogeographic significance (Lepidoptera: Hepialidae)

New species of Phassus Walker ghost moth from northeastern Mexico and notes on its biogeographic significance (Lepidoptera: Hepialidae)

Immulectin-2 from the ghost moth, Thitarodes xiaojinensis (Lepidoptera: Hepialidae), modulates cellular and humoral responses against fungal infection

C type-lectins constitute a large family of pattern recognition receptors, playing important roles in insect immune defenses. Thitarodes xiaojinensis larvae showed distinct immune features after Ophiocordyceps sinensis, Cordyceps militaris, or Beauveria bassiana infection. Based on transcriptome and immunoblot analysis, we found that immulectin-2 (IML2) was induced after T. xiaojinensis larvae were infected by C. militaris or B. bassiana but maintained at a low level after larvae injected with O. sinensis or Ringer's buffer. Recombinant IML2 (rIML2) could promote melanization, encapsulation, phagocytosis, and hemocyte aggregation in vitro. RNA interference with IML2 induced a significant reduction in the transcript levels of various antimicrobial peptides. Importantly, we found that the abundance of O. sinensis blastospores coated with rIML2 dramatically decreased in the host hemolymph. Overall, this study demonstrated that T. xiaojinensis IML2 modulates cellular and humoral responses to entomopathogenic fungi, broadening our view of the immune interaction between O. sinensis and its host.

A new species of Endoclita C. & R. Felder, 1874 ghost moth from Karimganj District of Assam, India, with comments on the genus in northeastern India, and clarification of E. signifer (Walker, 1856) (Lepidoptera: Hepialidae)

A new species of Endoclita C. & R. Felder, 1874 ghost moth from Karimganj District of Assam, India, with comments on the genus in northeastern India, and clarification of E. signifer (Walker, 1856) (Lepidoptera: Hepialidae)

Stage- and Rearing-Dependent Metabolomics Profiling of Ophiocordyceps sinensis and Its Pipeline Products

Simple SummaryThe caterpillar fungus O. sinensis is a historical ethnopharmacological commodity in China. The recent die-off of its wild population has raised the urgent task of ensuring the natural conservation of both the fungus and its host species Thitarodes. During the last decade, artificial cultivation of the caterpillar fungus has been established to supplement its declining natural colony. However, it is crucial to determine the nutritional background of the artificially reared cordyceps complex and its related products, i.e., mycelia and fruiting bodies reared on media. The current study aims to determine the comprehensive metabolic profiles of 17 treatments from 3 groups, including O. sinensis fungus, Thitarodes insect and cordyceps complex. We found that the metabolomics of O. sinensis-related products were mainly determined by the fruiting bodies rather than the culture methods. Our results suggest that artificially cultured fruiting bodies and cordyceps may have indistinguishable metabolite compositions as the natural ones. These results are highly intriguing in many aspects, from understanding the infection mechanism of caterpillar fungus, to commercial applications of caterpillar fungi quality authentication. It provides insights into the development of better alternatives for wild cordyceps and serves the future biological conservation of the fungal/insect species.Cordyceps, a parasitic complex of the fungus Ophiocordyceps sinensis (Berk.) (Hypocreales: Ophiocordycipitaceae) and the ghost moth Thitarodes (Lepidoptera: Hepialidae), is a historical ethnopharmacological commodity in China. Recently, artificial cultivation of Chinese cordyceps has been established to supplement the dwindling natural resources. However, much is unknown between the natural and cultivated products in terms of nutritional aspect, which may provide essential information for quality evaluation. The current study aims to determine the metabolic profiles of 17 treatments from 3 sample groups including O. sinensis fungus, Thitarodes insect and cordyceps complex, using Gas Chromatography - Quadrupole Time-of-Flight Mass Spectrometry. A total of 98 metabolites were detected, with 90 of them varying in concentrations among groups. The tested groups could be separated, except that fungal fruiting body was clustered into the same group as Chinese cordyceps. The main distinguishing factors for the groups studied were the 24 metabolites involved in numerous different metabolic pathways. In conclusion, metabolomics of O. sinensis and its related products were determined mainly by the fruiting bodies other than culture methods. Our results suggest that artificially cultured fruiting bodies and cordyceps may share indistinguishable metabolic functions as the natural ones.

Distinct Responses of Thitarodes xiaojinensis β-1,3-Glucan Recognition Protein-1 and Immulectin-8 to Ophiocordyceps sinensis and Cordyceps militaris Infection.

Melanization and encapsulation are prominent defense responses against microbes detected by pattern recognition receptors of their host insects. In the ghost moth Thitarodes xiaojinensis, an activated immune system can melanize and encapsulate the fungus Cordyceps militaris However, these responses were hardly detected in the host hemolymph postinfection of another fungus Ophiocordyceps sinensis The immune interaction between O. sinensis and the host remains largely unknown, which hinders the artificial cultivation of Chinese cordyceps. We found that T. xiaojinensis β-1,3-glucan recognition protein-1 (βGRP1) was needed for prophenoloxidase activation induced by C. militaris Failure of βGRP1 to recognize O. sinensis is a primary reason for the lack of melanization in the infected host. Lyticase or snailase treatment combined with binding and immunofluorescence detection showed the existence of a protective layer preventing the fungus from βGRP1 recognition. Coimmunoprecipitation and mass spectrometry analysis indicated that βGRP1 interacted with immulectin-8 (IML8) via binding to C. militaris IML8 promotes encapsulation. This study suggests the roles of T. xiaojinensis βGRP1 and IML8 in modulating immune responses against C. militaris Most importantly, the data indicate that O. sinensis may evade melanization by preventing βGRP1 recognition.

Gut Bacterial and Fungal Communities of the Wild and Laboratory-Reared Thitarodes Larvae, Host of the Chinese Medicinal Fungus Ophiocordyceps sinensis on Tibetan Plateau

Simple SummaryThe ghost moth, Thitarodes sp., is an obligate host of the most precious fungus Ophiocordyceps sinensis on Tibetan plateau. Artificial rearing of the ghost moth at low-altitude laboratory by mimicking the environment of the wild habitat for the cultivation of the Chinese cordyceps has been realized. However, the high mortality of ghost moth larvae by pathogens, low and slow infection, and mummification rate by O. sinensis still constrain the efficient cultivation of the Chinese cordyceps. Both larval gut microbiota and their exploitation in the Thitarodes artificial rearing system have attracted a renewed interest. In the present study, the gut bacterial and fungal communities of the wild and laboratory-reared populations were characterized using both culture-dependent and -independent approaches. The discovery of apparent microbial community shifts between the wild and laboratory-reared ghost moth larvae, many opportunistic pathogenic bacteria and fungi in the gut of the laboratory-reared ghost moth larvae, and the dominant bacteria enriched in the wild ghost moth provide interesting cues for selecting beneficial probiotic bacteria to improve the effectiveness of Thitarodes rearing system and the cultivation of the Chinese cordyceps.By employing a culture-dependent and -independent 16S rRNA and ITS gene high-throughput sequencing analyses, comprehensive information was obtained on the gut bacterial and fungal communities in the ghost moth larvae of three different geographic locations from high-altitude on Tibet plateau and from low-altitude laboratory. Twenty-six culturable bacterial species belonging to 21 genera and 14 fungal species belonging to 12 genera were identified from six populations by culture-dependent method. Carnobacterium maltaromaticum was the most abundant bacterial species from both the wild and laboratory-reared larvae. The most abundant OTUs in the wild ghost moth populations were Carnobacteriaceae, Enterobacteriaceae for bacteria, and Ascomycota and Basidiomycota for fungi. Larval microbial communities of the wild ghost moth from different geographic locations were not significantly different from each other but significant difference in larval microbial community was detected between the wild and laboratory-reared ghost moth. The larval gut of the wild ghost moth was dominated by the culturable Carnobacterium. However, that of the laboratory-reared ghost moth exhibited significantly abundant Wolbachia, Rhizobium, Serratia, Pseudomonas, and Flavobacterium. Furthermore, the larval gut of the wild ghost moth had a significantly higher abundance of Ophiocordyceps but lower abundance of Candida and Aspergillus than that of the laboratory-reared ghost moth.

<p><strong>Observations on <em>Phassus n-signatus </em>Weymer (Lepidoptera: Hepialidae) feeding on <em>Ligustrum lucidum</em> in a suburban habitat of Heredia, Costa Rica</strong></p>

Larval feeding by the Central American ghost moth Phassus n-signatus is recorded in the Asian tree Ligustrum lucidum for the first time. Larval feeding and feeding damage was observed on 15 plants in a suburban environment where L. lucidum is widely used as a residential hedge. Larvae feed under a web of silk and frass on callus wound tissue that grows back following removal of bark around the entrance to a short (less than 42 cm) tunnel where the larva resides when at rest. The diameter of host plant stems at the tunnel site ranged between 4–32 cm and most larval tunnels were found less than 1.5 m from the ground. The presence of P. n-signatus within a suburban residential area shows that this species can persist within human modified urbanized environments where there are sufficient host plants available, and ground conditions include decaying plant debris that would be necessary to support growth of early instars that are presumed to occupy this microhabitat as documented for other stem boring Hepialidae. Key words: callus feeding, Central America, ecology, host plant, larva, phytophagy, stem borer

Novel formulation development from Ophiocordyceps sinensis (Berk.) for management of high-altitude maladies.

Ophiocordyceps sinensis (Berk.) is a fungus closely related to medicinal mushroom, which belongs to the family Ophiocordycipitaceae. It is a well-known and rich herbal source of bioactive active constituents. The medicinal mushroom has garnered worldwide attention owing to its multifarious bioactivities. This mushroom grows on the larva of ghost moths (Hepialidae) and produces fruiting bodies, which serve as a vital natural source of medicine and supplementary diets. On account of the diverse pharmacological and bioactive constituents present in O. sinensis, it has been established as a potential antioxidant, anticancer, antibacterial, anti-proliferative, anti-inflammatory agent that has been successfully used for treating several health issues, including hypoxia-related problems encountered by mountaineers, pilgrims, tourists and soldiers occurring at high-altitude regions such as acute mountain sickness (AMS), high-altitude pulmonary edema (HAPE), high-altitude cerebral edema (HACE), frostbite, chilblains, hypothermia, etc. The most important pharmacologically active compounds present in the O. sinensis include nucleobases and its derivatives (adenosine, cordycepin, 3-deoxyadenosine, AMP, GMP, UMP, guanosine, uridine), polysaccharides (mannose, glucose, galactose, rhamnose, arabinose, xylose, galactose), proteins, peptides and steroids. This article focuses on the various research endeavors undertaken to scientifically establish the medicinal properties of O. sinensis, highlighting the various principally active compounds, their pharmacological action, drug designing and development and future perspective for various health benefits.

Infection of Ophiocordyceps sinensis Fungus Causes Dramatic Changes in the Microbiota of Its Thitarodes Host

The Chinese cordyceps is a unique and valuable parasitic complex of Thitarodes/Hepialus ghost moths and the Ophiocordyceps sinensis fungus for medicine and health foods from the Tibetan Plateau. During artificial cultivation of Chinese cordyceps, the induction of blastospores into hyphae is a prerequisite for mummification of the infected Thitarodes larvae. To explore the microbial involvement in the induction of mycelia-blastospore transition, the microbiota of the hemolymph and gut from Thitarodes xiaojinensis larvae with or without injected O. sinensis blastospores were investigated by culture-dependent and -independent methods. Twenty-five culturable bacterial species and 14 fungal species, together with 537 bacterial operational taxonomic units (OTUs) and 218 fungal OTUs, were identified from the hemolymph and gut of samples from five stages including living larvae without injected fungi (A) or with high blastospore load (B), mummifying larvae without mycelia coating (C), freshly mummifying larvae coated with mycelia (D), and completely mummified larvae with mycelia (E). Two culturable bacterial species (Serratia plymuthica, Serratia proteamaculans), and 47 bacterial and 15 fungal OTUs were considered as shared species. The uninfected larval hemolymph contained 13 culturable bacterial species but no fungal species, together with 164 bacterial and 73 fungal OTUs. To our knowledge, this is the first study to detect large bacterial communities from the hemolymph of healthy insect larvae. When the living larvae contained high blastospore load, the culturable bacterial community was sharply inhibited in the hemolymph but the bacterial and fungal community greatly increased in the gut. In general, high blastospore load increased bacterial diversity but sharply decreased fungal diversity in the hemolymph and gut by OTUs. The bacterial loads of four culturable species (Chryseobacterium sp., Pseudomonas fragi, S. plymuthica, S. proteamaculans) increased significantly and O. sinensis and Pseudomonas spp. became dominant microbes, when the infected larvae became mummified, indicating their possible involvement in the larval mummification process. The discovery of many opportunistic pathogenic bacteria in the hemolymph of the healthy larvae, the larval microbial diversity influenced by O. sinensis challenge and the involvement of dominant bacteria during larval mummification process provide new insight into the infection and mummification mechanisms of O. sinensis in its Thitarodes hosts.