In-depth transcriptome and physiological function analysis reveals the toxicology of sodium fluoride in the fall webworm Hyphantria cunea.

Fluoride is an environmental pollutant that severely injures various organisms in ecosystems. Herein, the non-target organism, fall webworm (Hyphantria cunea), was used to determine the toxicological mechanism of NaF exposure. In this study, H. cunea exposed to NaF showed significant declines in growth and reproduction. The authors conducted RNA sequencing on adipose bodies and midgut tissues from NaF-exposed H. cunea larvae to uncover the toxicological mechanisms. The results showed that extracellular matrix-receptor interaction, pentose and glucuronate interconversions, fatty acid biosynthesis, and ferroptosis might contribute to NaF stress. NaF significantly decreased the antioxidant level, nitrous oxide synthase activity, and NO content, while significantly increasing lipid peroxidation. NaF induced significant changes in the expression of energy metabolism genes. However, the triglyceride content was significantly decreased and the lipase enzyme activity was significantly increased. Moreover, the expression levels of light and heavy chains of ferritin were inhibited in NaF-exposed H. cunea. NaF caused ferritin Fe2+overload in FerHCH1 and FerLCH knockdown H. cunea larvae, activated reactive oxygen species, and reduced the total iron content, eventually increasing the mortality H. cunea larvae. This study identified the toxicological mechanisms of NaF in lipid synthesis and energy metabolism in H. cunea, providing a basis for understanding the molecular mechanisms of NaF toxicity and developing pollution control strategies.

Molecular basis of camphor repellency in Hyphantria cunea.

The plant-derived camphor has been used as a natural insect repellent against various insects for >500 years. However, the repellency mechanism behind camphor remains less understood. In this study, we aimed to identify the camphor receptor in Hyphantria cunea by deorphanizing 7 odorant receptors (ORs). The results showed that HcunOR46 is narrowly tuned to Camphor and is only conserved within the family Noctuidae. Further analysis through behavioral and electroantennograms (EAG) assays indicated that H. cunea adults are more sensitive to camphor than larvae, both behaviorally and electrophysiologically. This difference may be due to the lower expression of HcunOR46 at the larval stage. Additionally, a feeding assay indicated that camphor repellency could be related to camphor toxicity to larvae, with the lethal concentration 50 (LC50) value of 69.713 µg/µL. These results suggest that H. cunea may detect camphor through a distinct olfactory pathway from Culicinae mosquitos, providing a novel camphor-based pest management strategy for H. cunea.

Molecular Basis of CO2 Sensing in Hyphantria cunea.

Carbon dioxide (CO2) released by plants can serve as a cue for regulating insect behaviors. Hyphantria cunea is a widely distributed forestry pest that may use CO2 as a cue for foraging and oviposition. However, the molecular mechanism underlying its ability to sense CO2 has not been elucidated. Our initial study showed that CO2 is significantly attractive to H. cunea adults. Subsequently, 44 H. cunea gustatory receptors (GRs) were identified using transcriptome data, and 3 candidate CO2 receptors that are specifically expressed in the labial palps were identified. In vivo electrophysiological assays revealed that the labial palp is the primary organ for CO2 perception in H. cunea, which is similar to findings in other lepidopteran species. By using the Xenopus oocyte expression system, we showed that the HcunGR1 and HcunGR3 co-expressions produced a robust response to CO2, but HcunGR2 had an inhibitory effect on CO2 perception. Finally, immunohistochemical staining revealed sexual dimorphism in the CO2-sensitive labial pit organ glomerulus (LPOG). Taken together, our results clarified the mechanism by which H. cunea sense CO2, laying the foundation for further investigations into the role of CO2 in the rapid spread of H. cunea.

Insecticidal activities of the local entomopathogenic nematodes and cell-free supernatants from their symbiotic bacteria against the larvae of fall webworm, Hyphantriacunea.

The fall webworm (FWW), Hyphantria cunea Drury (Lepidoptera: Erebidae), is an invasive and polyphagous insect pest of many economically important crops such as hazelnuts, apple, and mulberry. Recently, there have been an increasing number of reports about the damaging activities of FWW from hazelnut growing areas of Turkey indicating that currently existing control methods fail to satisfy the expectations of growers. Entomopathogenic nematodes (EPNs) in the Steinernematidae and Heterorhabditidae (Nematoda: Rhabditida) families and the symbiotic bacteria they carry in their intestine have a great potential for the management of many agriculturally important pests. In this study, the symbiotic bacteria of local EPN species (Heterorhabditis bacteriophora AVB-15, Steinernema feltiae KCS-4S, and Steinernema bicornotum MGZ-4S) recovered from the central Anatolia region was characterized using recA gene region as Photorhabdus luminescens, Xenorhabdus bovienii and Xenorhabdus budapestensis. The contact (25, 50, 100, 200 IJs/Petri) and oral efficacies of the infective juveniles (IJs) (25, 50, 100, 200 IJs/leaf) of these EPN isolates determined on 3rd/4th instar larvae, and cell-free supernatants from the identified symbiotic bacteria were evaluated separately on the 3rd and 4th larval instars of FWW in Petri dish environment under laboratory conditions (25 ± 1 °C, 60% of RH). In the Petri dish bioassays of EPN species, the most pathogenic isolate at the 1st DAT and 4th DAT was S. feltiae which caused 50% mortality at the highest concentration (200 IJs/Petri) and the highest mortality rate (97.5%) were achieved at 4th DAT by H. bacteriophora AVB-15 isolate. Surprisingly, the mortality rates were generally higher at the lowest concentrations and 82.5% mortality were reached 4th DAT by S. bicornotum at the lowest concentration (25 IJs/leaf) in the leaf bioassays. Mortality rates were higher in both Petri dish and filter paper efficacies of cell-free supernatants at the 2nd DAT and the highest mortality (87.5%) was reached in the contact efficacy studies when applied X. bovienii KCS-4S strain. The results suggest that the tested EPN species and CFSs have good potential for biological control of the larvae of FWW and can contribute to the IPM programs of FWW. However, the efficacy of both IJs of EPNs and CFSs of their symbiotic bacteria on larvae of FWW requires further studies to verify their efficiency in the field.

First record of Oscheius myriophilus () (Rhabditida: Rhabditidae) from Iran; and its efficacy against two economic forest trees pests, Cydalima perspectalis (Walker, 1859) (Lepidoptera: Crambidae) and Hyphantria cunea () (Lepidoptera: Erebidae) in laboratory condition.

The box tree moth (BTM), Cydalima perspectalis and the fall webworm (FWW), Hyphantria cunea are two invasive pests of forest trees that have been recorded from Hyracinan forests in north Iran for the first time in 2016 and 2002, respectively. In a search for tentative native entomopathogenic nematode species (EPNs) with potential biocontrol ability against lepidopteran pests of forest trees in north Iran, Oscheius myriophilus was isolated by soil-baiting method from forests of Amlash in the east of Gilan province. The Iranian isolate of this species is characterized by 870-1,247 µm long hermaphrodites having 17-20 µm long stoma, vulva at 45.5-53.1% of body length, tail 90-126 µm long, common males with 38-49 µm long spicules and 583-791 µm long dauer larvae. Compared to the type and other populations, no remarkable differences were observed for this population. The phylogenetic affinities of this isolate with other rhabditid nematodes were studied using partial sequences of small, and the D2-D3 expansion segments of the large subunit ribosomal DNA (SSU and D2-D3 LSU rDNA). This is the first record of the species from Iran. The pathogenicity of Iranian isolate of O. myriophilus was evaluated on the larvae of two important aforementioned forest trees pests, BTM and FWW under laboratory conditions. The results indicated that the suspension of 500 infective juveniles per ml (IJs/ml) of the nematode was the most effective treatment on fifth instar larvae of BTM and FWW, causing 100 and 95% mortality after 48 h, respectively. The lethal concentration 50 (LC50) values of the nematode were 74.5, 152.7, 99.9, and 197.3 IJs/ml on fifth and fourth instar larvae of BTM, and fifth and fourth instar larvae of FWW, respectively, after 48 h at 25°C and 60% relative humidity. Together, present results corroborated the efficacy of the Iranian isolate of Oscheius myriophilus for biocontrolling of BTM and FWW in laboratory conditions.

Genome of the webworm Hyphantria cunea unveils genetic adaptations supporting its rapid invasion and spread.

BACKGROUND: The fall webworm Hyphantria cunea is an invasive and polyphagous defoliator pest that feeds on nearly any type of deciduous tree worldwide. The silk web of H. cunea aids its aggregating behavior, provides thermal regulation and is regarded as one of causes for its rapid spread. In addition, both chemosensory and detoxification genes are vital for host adaptation in insects. RESULTS: Here, a high-quality genome of H. cunea was obtained. Silk-web-related genes were identified from the genome, and successful silencing of the silk protein gene HcunFib-H resulted in a significant decrease in silk web shelter production. The CAFE analysis showed that some chemosensory and detoxification gene families, such as CSPs, CCEs, GSTs and UGTs, were expanded. A transcriptome analysis using the newly sequenced H. cunea genome showed that most chemosensory genes were specifically expressed in the antennae, while most detoxification genes were highly expressed during the feeding peak. Moreover, we found that many nutrient-related genes and one detoxification gene, HcunP450 (CYP306A1), were under significant positive selection, suggesting a crucial role of these genes in host adaptation in H. cunea. At the metagenomic level, several microbial communities in H. cunea gut and their metabolic pathways might be beneficial to H. cunea for nutrient metabolism and detoxification, and might also contribute to its host adaptation. CONCLUSIONS: These findings explain the host and environmental adaptations of H. cunea at the genetic level and provide partial evidence for the cause of its rapid invasion and potential gene targets for innovative pest management strategies.

Exploring the Dynamic Invasion Pattern of the Black-Headed Fall Webworm in China: Susceptibility to Topography, Vegetation, and Human Activities.

The fall webworm (FWW), H. cunea (Drury) (Lepidoptera: Erebidae: Arctiidae), is an extremely high-risk globally invasive pest. Understanding the invasion dynamics of invasive pests and identifying the critical factors that promote their spread is essential for devising practical and efficient strategies for their control and management. The invasion dynamics of the FWW and its influencing factors were analyzed using standard deviation ellipse and spatial autocorrelation methods. The analysis was based on statistical data on the occurrence of the FWW in China. The dissemination pattern of the FWW between 1979 and 2022 followed a sequence of "invasion-occurrence-transmission-outbreak", spreading progressively from coastal to inland regions. Furthermore, areas with high nighttime light values, abundant ports, and non-forested areas with low vegetation cover at altitudes below 500 m were more likely to be inhabited by the black-headed FWW. The dynamic invasion pattern and the driving factors associated with the fall webworm (FWW) provide critical insights for future FWW management strategies. These strategies serve not only to regulate the dissemination of insects and diminish migratory tendencies but also to guarantee the implementation of efficient early detection systems and prompt response measures.

Niche and Range Shifts of the Fall Webworm (Hyphantria cunea Dury) in Europe Imply Its Huge Invasion Potential in the Future.

The fall webworm (Hyphantria cunea Dury) has a strong impact on agricultural systems in Europe. However, its invasive potential, which was inherited from its native niche in North America, remains unknown. Here, we investigated the climatic niche and range shifts of the fall webworm in Europe and compared them with those in native North America, then assessed the worms' invasive potential in Europe. Compared with the fall webworm in Europe, those in North America survived in more diverse climatic conditions, which was closely associated with their broader niche and larger potential ranges in Europe. If the fall webworm in Europe could exploit the native niche inherited from those in North America to adapt to climatic conditions in Europe, their potential ranges in Europe could be 5.5-fold those based on the niche as introduced in Europe. The potentially unfilled ranges of the fall webworm in Europe were mainly detected in vast regions of Europe, excluding Norway, Sweden, Finland, North Russia, Hungary, Croatia, Romania, and Ukraine, suggesting that, without strict control, these vast regions might be preferably invaded by the fall webworm in Europe in the future. Therefore, strict control against its invasion is needed. Given that small niche shifts in this invasive insect could result in large range shifts, the niche shifts represent a more sensitive indicator of invasion risk than range shifts.

Bottom-up and top-down pressures mediate competition between two generalist insects.

The effects of competition can have far-reaching consequences for individuals, populations, and communities and therefore we should strive toward a deeper understanding of competitive interactions. In some cases, dietary generalists may be predicted to experience weak competition effects because of their ability to use a wide range of host plants. However, competition between insects frequently occurs indirectly, which can hinder insects' abilities to avoid competitive interactions. Therefore, competition may be as strong among dietary generalists as among dietary specialists. Yet competition between insects that are dietary generalists is infrequently studied. We tested for evidence of competitive interactions between two common, temporally separated, generalist insects: the western tent caterpillar (Malacosoma californicum), which feeds early in the season, and the fall webworm (Hyphantria cunea), which feeds later in the season. Both species frequently use a common host plant species (chokecherry) as a preferred host at our field sites. We tested the relative strength of bottom-up effects resulting from competitive interactions between these two generalists with laboratory-rearing trials at the relevant time of year for each insect. We recorded three common fitness measures (development time, pupal mass, and survival) for caterpillars reared on chokecherry with no damage from either of our focal species, with tent caterpillar damage, and with fall webworm damage. To test the strength of top-down pressures on fall webworm larval fitness and any potential interactions with bottom-up effects, we reared larvae in the field either exposed to or protected from predators on host plants that either did or did not have tent caterpillars feeding on them earlier in the season. We found evidence of bottom-up fitness effects on tent caterpillars and top-down and bottom-up fitness effects on fall webworms confirming that tent caterpillars and fall webworms compete indirectly. Tent caterpillars had lower pupal mass when reared on leaves from shrubs damaged by fall webworms. Fall webworms had lower pupal mass and longer development time when reared on leaves from shrubs damaged by tent caterpillars. In field trials, fall webworms reared on shrubs damaged by tent caterpillars had a lower survival and pupal mass. We show evidence of indirect competition in temporally separated generalists through leaf quality (bottom-up effects) and natural enemies (top-down effects).

Bursicon receptor gene HLGR2 as a potential RNA interference target for control of the fall webworm Hyphantria cunea.

BACKGROUND: Insect G protein-coupled receptors (GPCRs) have been identified as a new generation of attractive targets for RNA interference (RNAi)-based pest control. A functional study of the leucine-rich repeat-containing (LGR2) gene in Hyphantria cunea (HLGR2) was performed to examine whether it can be used in the molecular control of this notorious pest. RESULTS: The complementary DNA (cDNA) sequence and deduced amino acids of HLGR2 were obtained and analyzed in the present study. HLGR2 is a typical GPCR and shows high structural and sequence similarity with other insect LGR2 proteins. The spatiotemporal expression profiles of HLGR2 showed that HLGR2 was highly expressed at the egg stage and tissues of head and silk gland. After RNAi of HLGR2, distinct phenotypes were observed when HLGR2 expression was suppressed, indicating that HLGR2 is essential in pupation and eclosion. HLGR2 RNAi led to a low pupation rate (45.00%), body malformation, abnormal wing expansion, failed cuticle melanization (63.33%), and high mortality rate (48.33%). Furthermore, we identified eight genes that are regulated by HLGR2. The expression of these eight genes was induced by the HLGR2 signaling pathway and correlated well with cuticle sclerotization. Unlike LGR2 in other insect species, HLGR2 was found to play a crucial role in the control of H. cunea during ecdysis and postecdysial stages. CONCLUSION: HLGR2 is essential for the growth and development and wing expansion and maturation in H. cunea, suggesting HLGR2 is a promising candidate for application in RNAi-based control of this notorious agriculture-forest pest. © 2021 Society of Chemical Industry.

Repurposing a traditional Japanese method of pest control for wintering pine moths, Komo-trap, for use against summer and winter populations of fall webworms.

BACKGROUND: The fall webworm, Hyphantria cunea Drury (Lepidoptera: Erebidae), is a widespread invasive species. It is native to North America, ranging from southern Canada to northern Mexico. During and after the 1940s, this pest was accidentally introduced in many parts of Europe and Asia. It has now spread to more than 30 countries. The larvae feed on leaves of a wide range of tree species, including ones used as street trees in cities, causing an increase in urban management cost. Although several pest management methods have been employed, pest damage continues especially in newly invaded areas. In this study, we examined the effect and cost-effectiveness of the komo-trap, traditionally used in Japan to reduce the population of larvae of the pine moth Dendrolimus spectabilis Butler (Lepidoptera: Lasiocampidae). This trap, which is safe for people and ecosystems, has not yet been applied to trap the fall webworm. METHODS: In two seasons of 2017, we set komo-traps on street trees in Hakodate City, Japan. We counted the numbers of captured fall webworms compared with controls. We also monitored other species to evaluate any nontarget effects of the trap. RESULTS: One komo, the material cost of which is about 1.10 USD, captured 43.8 fall webworms on average in summer and 27.2 in the fall. The values were significantly larger than those of the controls, which were 0.07 in summer and 0.14 in winter. Bycatch of other species was minimal in summer, whereas in the fall one komo, on average, caught 10.7 woodlice Porcellio sp. or spp. (Isopoda: Porcellionidae). DISCUSSION: The komo-trap is effective in capturing fall webworm. The cost performance of the trap is very favorable, and the nontarget effects can be reduced by using the trap in summer only. The komo-trap would complement other control methods such as tree pruning. Because its cost is low, we recommend that the komo-trap be introduced as a larger-scale trial.

Biocontrol potential of Beauveria bassiana and Metarhizium anisopliae isolates from Turkey against Hyphantria cunea (Drury) (Lepidoptera: arctiidae) Larvae under laboratory and field conditions

Hyphantria cunea is one of the most important pest insects causing significant damage in many plant species. The present study aimed to evaluate the insecticidal activity of Turkey isolates of Beauveria bassiana and Metarhizium anisopliae against H. cunea larvae under laboratory and field conditions. B. bassiana isolates YK16, YK23 and YK26, and M. anisopliae isolates YK41 and YK45 were sprayed onto the larvae of H. cunea at the respective doses of 1x105 and 1x106 conidia mL-1 and monitored for seven days. Fungal isolates, bio-insecticide Bacillus thuringiensis and chemical insecticide diflubenzuron were used in field studies. Trials were carried out in five replications. All B. bassiana isolates caused 100% mortality on first instar larvae in laboratory trials. Mortality ratios ranged from 100 to 96% on second instar larvae. M. anisopliae isolates YK45 and YK41 caused 88 and 84%, and 81.33 and 77.11% mortalities for the first and second instar larvae, respectively. The mortality rates fluctuated between 91.78-72.89% for B. bassiana on third instar larvae in laboratory conditions. However, M. anisopliae isolates YK45 and YK41 caused 77.11 and 60.22% mortality on third instar larvae, respectively. In the field trials, B. bassiana YK23 displayed promising insecticidal activity with 80.60% mortality on second instar larvae of H. cunea. Other isolates as well caused mortalities ranging from 60.77 to 49.55%. The results revealed that some isolates of B. bassiana and M. anisopliae have potential to control H. cunea larvae. However, additional detailed studies need to be carried out to increase their effectiveness in field conditions.

The complete mitochondrial genome of the fall webworm, Hyphantria cunea (Lepidoptera: Arctiidae).

The complete mitochondrial genome (mitogenome) of the fall webworm, Hyphantria cunea (Lepidoptera: Arctiidae) was determined. The genome is a circular molecule 15 481 bp long. It presents a typical gene organization and order for completely sequenced lepidopteran mitogenomes, but differs from the insect ancestral type for the placement of tRNA(Met). The nucleotide composition of the genome is also highly A + T biased, accounting for 80.38%, with a slightly positive AT skewness (0.010), indicating the occurrence of more As than Ts, as found in the Noctuoidea species. All protein-coding genes (PCGs) are initiated by ATN codons, except for COI, which is tentatively designated by the CGA codon as observed in other lepidopterans. Four of 13 PCGs harbor the incomplete termination codon, T or TA. All tRNAs have a typical clover-leaf structure of mitochondrial tRNAs, except for tRNA(Ser)(AGN), the DHU arm of which could not form a stable stem-loop structure. The intergenic spacer sequence between tRNA(Ser)(AGN) and ND1 also contains the ATACTAA motif, which is conserved across the Lepidoptera order. The H. cunea A+T-rich region of 357 bp is comprised of non-repetitive sequences, but harbors several features common to the Lepidoptera insects, including the motif ATAGA followed by an 18 bp poly-T stretch, a microsatellite-like (AT)(8) element preceded by the ATTTA motif, an 11 bp poly-A present immediately upstream tRNA(Met). The phylogenetic analyses support the view that the H. cunea is closerly related to the Lymantria dispar than Ochrogaster lunifer, and support the hypothesis that Noctuoidea (H. cunea, L. dispar, and O. lunifer) and Geometroidea (Phthonandria atrilineata) are monophyletic. However, in the phylogenetic trees based on mitogenome sequences among the lepidopteran superfamilies, Papillonoidea (Artogeia melete, Acraea issoria, and Coreana raphaelis) joined basally within the monophyly of Lepidoptera, which is different to the traditional classification.