Asian Gypsy Moth Research Articles

Monitoring the occurrence of Asian gypsy moth(Lymantria dispar asiatica)with sex pheromone-baited trap in five types of forest stands

Monitoring the occurrence of Asian gypsy moth(Lymantria dispar asiatica)with sex pheromone-baited trap in five types of forest stands

Global establishment threat from a major forest pest via international shipping: Lymantria dispar

The global shipping network is widely recognised as a pathway for vectoring invasive species. One species of particular concern is Lymantria dispar (gypsy moth). Two subspecies, L. d. asiatica and L. d. japonica (herein referred to as Asian Gypsy Moth - AGM) are of considerable concern as ships arriving to a number of countries have been found carrying AGM egg masses. However, ships carrying AGM eggs can only threaten a country at ports located in a climatically suitable region. We present a CLIMEX model of climate suitability and combine this with international shipping to estimate the global threat from AGM. We find that for the USA more than half of international ships (approximately 18,000 ships) arrive to climatically suitable ports. Other countries with a large number of ships arriving to ports with suitable climates include Canada and Brazil. This is the first global analysis of the invasion threat from AGM, and we recommend countries focus AGM-inspection programs towards ships arriving at ports found within climatically suitable regions.

Identification of Japanese Lymantria species (Lepidoptera: Lymantriidae) based on DNA sequences

We examined the utility of the cytochrome c oxidase subunit I (COI) and II (COII) genes of mitochondrial DNA (mtDNA) as a tool to identify nine Japanese Lymantria species, including four Asian gypsy moth species [Lymantria dispar japonica (Motschulsky), Lymantria umbrosa (Butler), Lymantria albescens Hori and Umeno, and Lymantria postalba Inoue]. In phylogenetic trees constructed for the COI and COII genes using the maximum likelihood methods, we could identify seven out of the nine Lymantria species (L. albescens and L. postalba could not be identified). These results suggest that the DNA sequences of the COI and COII genes are useful for identifying Japanese Lymantria species.

Pheromone-binding proteins in the Asian gypsy moth females, Lymantria dispar, recognizing the sex pheromone and plant volatiles.

Lepidopterans are known to have different pheromone-binding proteins with differential expression patterns that facilitate specific signal transduction of semiochemicals. Two PBPs of the Asian gypsy moth, Lymantria dispar, were reported to express in both females and males, but their physiological functions were unknown. Results showed that LdisPBP1 and LdisPBP2 were expressed in the sensilla trichodea of males and the s. trichodea and s. basiconica of females. When LdisPBP1 gene was targeted by RNA interference (RNAi) in males, the expression of LdisPBP1 and LdisPBP2 decreased by 69 and 76%, respectively, and when LdisPBP2 gene was targeted by RNAi, they decreased by 60 and 42%, respectively. In females, after treatment with LdisPBP1 dsRNA, LdisPBP1 and LdisPBP2 levels were reduced by 26 and 69%, respectively, and LdisPBP2 dsRNA reduced the relative expression of them by 4 and 62%, respectively. The expression of LdisPBP1 and LdisPBP2 was interdependent. Electroantennogram (EAG) recordings showed that LdisPBPs participate in the recognition of the sex pheromone in males, and the sex pheromone and plant volatiles in females. The function of LdisPBPs represents the sex-specific roles.

Synergic effect of sex pheromone (7R,8S)-cis-7,8-epoxy-2-methyloctadec-17-ene on (+)-disparlure of the Asian gypsy moth, Lymantria dispar, in Korea

field attraction test of (7R,8S)-cis-7,8-epoxy-2-methyloctadec-17-ene (= 7R,8S-epo-2me-17ene-18Hy), a trace component of the sex pheromone gland of the Asian gypsy moth, (Lymantria dispar), traps that were baited with the trace component attracted more L. dispar than traps baited with (7S,8R)-cis-7,8-epoxy-2-methyloctadec-17-ene (= 7S,8R-epo-2me-17ene-18Hy) and unbaited traps. However, traps baited with 7R,8S-epo-2me-17ene-18Hy were less attractive than traps baited with (+)-disparlure [= (7R,8S)-cis-7,8-epoxy-2-methyloctadecane], the sex pheromone of L. dispar. Combination tests with (+)-disparlure, 7R,8S-epo-2me-17ene-18Hy, and 7S,8R-epo-2me-17ene-18Hy revealed that 7R,8S-epo-2me-17ene-18Hy acted synergistically with (+)-disparlure.

Rapid identification of the Asian gypsy moth and its related species based on mitochondrial DNA.

The gypsy moth—Lymantria dispar (Linnaeus)—is a worldwide forest defoliator and is of two types: the European gypsy moth and the Asian gypsy moth. Because of multiple invasions of the Asian gypsy moth, the North American Plant Protection Organization officially approved Regional Standards for Phytosanitary Measures No. 33. Accordingly, special quarantine measures have been implemented for 30 special focused ports in the epidemic areas of the Asian gypsy moth, including China, which has imposed great inconvenience on export trade. The Asian gypsy moth and its related species (i.e., Lymantria monocha and Lymantria xylina) intercepted at ports are usually at different life stages, making their identification difficult. Furthermore, Port quarantine requires speedy clearance. As such, it is difficult to identify the Asian gypsy moth and its related species only by their morphological characteristics in a speedy measure. Therefore, this study aimed to use molecular biology technology to rapidly identify the Asian gypsy moth and its related species based on the consistency of mitochondrial DNA in different life stages. We designed 10 pairs of specific primers from different fragments of the Asian gypsy moth and its related species, and their detection sensitivity met the need for rapid identification. In addition, we determined the optimal polymerase chain reaction amplification temperature of the 10 pairs of specific primers, including three pairs of specific primers for the Asian gypsy moth (L. dispar asiatic), four pairs of specific primers for the nun moth (L. monocha), and three pairs of specific primers for the casuarina moth (L. xylina). In conclusion, using our designed primers, direct rapid identification of the Asian gypsy moth and its related species is possible, and this advancement can help improve export trade in China.

Costs and benefits of a leaf beetle Integrated Pest Management (IPM) program. I. Modelling changes in wood volume yields from pest management

ABSTRACTThe Tasmanian leaf beetle Paropsisterna bimaculata is a species native to Tasmania that can cause severe defoliation of eucalypt plantations. High populations of P. bimaculata, capable of causing severe defoliation if unmanaged, can periodically occur through a substantial proportion of a plantation rotation. Some exotic insect pests not yet established in Australia, most notably Asian Gypsy Moth (Lymantria dispar), share the similar characteristic of imposing a threat for a substantial proportion of a plantation rotation. Managing such a threat is operationally complex and requires committing adequate resources to sustain a robust management system. However, the costs and benefits to the plantation owner of sustaining such a management system have not been quantified. The first step of such an analysis is to quantify the gains in additional wood yields from management that protects against severe defoliation of plantations.Forestry Tasmania has conducted considerable research to develop an Integrated Pest Management (IPM) program to protect its plantations from damaging populations of P. bimaculata, and has been using this leaf beetle IPM program since the early 2000s. The program has generated many years of operational records from leaf beetle population monitoring. These historical population monitoring data coupled with knowledge of the impact of defoliation by leaf beetles on tree growth were used to estimate how much wood volume has been saved by protecting the Forestry Tasmania plantation estate from damaging large leaf beetle populations over an entire rotation. The probability of above-threshold leaf beetle populations (defined as populations that cause severe defoliation, >50%, if uncontrolled) could be predicted from (1) plantation age (the likelihood of above-threshold populations peaks at age 4–5 years and declines to a low value by age 12 years), and (2) site-level leaf beetle risk (sites of high leaf beetle risk were at higher elevation and closer to native grassland than sites of low leaf beetle risk). Based on these relationships, the occurrence of above-threshold leaf beetle populations was predicted in Forestry Tasmania’s 50 000 ha plantation estate between 2003 and 2034. The leaf beetle IPM program, through controlling these above-threshold populations, was then predicted to be able to avert losses of 2.18 million m3 (9.4% of merchantable wood volume).

Characterization and Expression Profiles of Juvenile Hormone Epoxide Hydrolase From Lymantria dispar (Lepidoptera: Lymantridae) and RNA Interference by Ingestion

Juvenile hormone epoxide hydrolase (JHEH) is an important enzyme in the degradation pathways of juvenile hormone (JH) in insects. It converts JH to JH diol and hydrolyses JH acid to JH acid diol. JHEH titers regulate the entire process of insect development. In this study, full length ldjheh cDNA (2101 bp) was cloned from the Asian gypsy moth Lymantria dispar (L.; Lepidoptera: Lymantridae), and provisionally designated ldjheh1. LdJHEH1 was characterized by predicted molecular weight of 52.64 kDa, theoretical isoelectric points of 6.87 and contains a transmembrane domain at the N-terminus. The transcriptional profiles of ldjheh1 were detected by qRT-PCR. The ldjheh1 was found to be expressed throughout all developmental stages with maximum expression levels occurring in fourth instar larvae. The ldjheh1 mRNA was detected in the heads, thoraces, and abdomens of gypsy moth larvae on day 2 of the third instar. The ldjheh1 was also detected in bodies of third instar larvae stage, with the highest peaks occurring at 24 h after ecdysis. The ldjheh1 gene was successfully knocked down by oral delivery dsRNA in the third instar larvae of L. dispar. The dsRNA targeting ldjheh1 was produced in vitro. Ingesting dsRNA for ldjheh1 only slightly delayed larval development.

Assessing the potential of genotyping‐by‐sequencing‐derived single nucleotide polymorphisms to identify the geographic origins of intercepted gypsy moth (Lymantria dispar) specimens: A proof‐of‐concept study

AbstractForest invasive alien species are a major threat to ecosystem stability and can have enormous economic and social impacts. For this reason, preventing the introduction of Asian gypsy moths (AGM; Lymantria dispar asiatica and L. d. japonica) into North America has been identified as a top priority by North American authorities. The AGM is an important defoliator of a wide variety of hardwood and coniferous trees, displaying a much broader host range and an enhanced dispersal ability relative to the already established European gypsy moth (L. d. dispar). Although molecular assays have been developed to help distinguish gypsy moth subspecies, these tools are not adequate for tracing the geographic origins of AGM samples intercepted on foreign vessels. Yet, this type of information would be very useful in characterizing introduction pathways and would help North American regulatory authorities in preventing introductions. The present proof‐of‐concept study assessed the potential of single nucleotide polymorphism (SNP) markers, obtained through genotyping by sequencing (GBS), to identify the geographic origins of gypsy moth samples. The approach was applied to eight laboratory‐reared gypsy moth populations, whose original stocks came from locations distributed over the entire range of L. dispar, comprising representatives of the three recognized subspecies. The various analyses we performed showed strong differentiation among populations (FST ≥ 0.237), enabling clear distinction of subspecies and geographic variants, while revealing introgression near the geographic boundaries between subspecies. This strong population structure resulted in 100% assignment success of moths to their original population when 2,327 SNPs were used. Although the SNP panels we developed are not immediately applicable to contemporary, natural populations because of distorted allele frequencies in the laboratory‐reared populations we used, our results attest to the potential of genomewide SNP markers as a tool to identify the geographic origins of intercepted gypsy moth samples.

Comparing the Expression of Olfaction-Related Genes in Gypsy Moth (Lymantria dispar) Adult Females and Larvae from One Flightless and Two Flight-Capable Populations

In insects, flight and sophisticated olfactory systems go hand in hand and are essential to survival and evolutionary success. Females of many Lepidopteran species have secondarily lost their flight ability, which may lead to changes in the olfactory capabilities of both larval and adult stages. The gypsy moth, Lymantria dispar, an important forest pest worldwide, is currently undergoing a diversification process with three recognized subspecies: the Asian gypsy moth (AGM) L. dispar asiatica, the Japanese gypsy moth L. dispar japonica (JGM) and the European gypsy moth (EGM) L. dispar dispar. Females of EGM populations from North America have lost their flight capacity whereas the JGM and AGM females are flight capable, making this an ideal system to investigate the relationship between flight and olfaction. We used next-generation sequencing to obtain female antennal and larval head capsule transcriptomes in order to (i) investigate the differences in expression of olfaction-related genes among populations; (ii) identify the most similar protein sequences reported for other organisms through a BLAST search, and (iii) establish the phylogenetic relationships of these sequences with respect to other insect species. Using this approach, we identified 115 putative chemosensory genes belonging to five families of olfaction-related genes. A principal component analysis revealed that the gene expression patterns of female antennal transcriptomes from different subspecies were more similar among them than to the larval head capsules of their respective subspecies supporting strong chemosensory differences between the two developmental stages. An analysis of the shared and exclusively expressed genes for three populations shows no evidence that loss of flight affects the number or type of genes being expressed. These results indicate either that (a) loss of flight does not impact the olfactory gene repertoire or that (b) the secondary loss of flight in the American EGM populations may be too recent to have caused major changes in the genes being expressed. However, we found higher expression values for IRs, OBPs GRs and ORs in EGM females, suggesting that differences in transcription rates could be an adaptation of flightless females to their chemical environment. Differences in olfactory genes and their expression in the

Effects of Temperature on Development of Lymantria dispar asiatica and Lymantria dispar japonica (Lepidoptera: Erebidae).

Periodic introductions of the Asian subspecies of gypsy moth, Lymantria dispar asiatica Vnukovskij and Lymantria dispar japonica Motschulsky, in North America are threatening forests and interrupting foreign trade. Although Asian gypsy moth has similar morphology to that of European and North American gypsy moth, it has several traits that make it a greater threat, the most important being the flight capability of females. Asian gypsy moth is not yet established in North America; however, infestations have been detected multiple times in Canada and the United States. To facilitate detection and eradication efforts, we evaluated the effect of a range of temperatures on development time, survivorship, and fertility of eight populations of Asian gypsy moth. There were significant impacts of temperature and population on these life history characteristics. The larval developmental rate increased with temperature until it reached an optimum at 29 °C. Larvae experienced significant molting problems at the highest and lowest temperatures tested (10 °C and 30 °C). At 30 °C, female fitness was markedly compromised, as evidenced by reduced fecundity and fertility. This suggests that development and survival of Asian gypsy moth may be limited by summer temperature extremes in the Southern United States. We also determined the degree-day requirements for two critical life stages and two populations of Asian gypsy moth, which represent the extremes in latitude, to predict the timing for biopesticide application and adult trap deployment. Our data will benefit pest managers in developing management strategies, pest risk assessments, and timing for implementation of management tactics.

Risk analysis of the invasion pathway of the Asian gypsy moth: a known forest invader

Risk is defined with many minor variations in the biological literature. Common to most definitions are the following elements: the probability of a future event; and the consequences of the event, usually with respect to some predefined human value. Risk analysis includes elements of risk assessment (quantification of risk), uncertainty (of the event and its consequences), risk management (reducing risk to an acceptable level), and development of policy to balance finite resources with uncertainty and risk tolerance. When biological invasion and its risk are jointly examined, it is common that the consequences of invasion are not explicitly quantified, but understood to be sufficiently negative that it must be minimized to the extent possible. Risk analysis then becomes quantification of the probabilities of an introduction (event) and that the introduction leads to establishment, and the uncertainty of those probabilities. I describe a risk analysis framework for the Asian gypsy moth—a known invader—in its pathway. The framework uses the available information regarding the transportation route of the vector (ships), and a phenology model that estimates vector contamination (propagule size), the probability of introduction, and the probability of initial establishment given an introduction. Reducing propagule pressure is arguably the most important factor in reducing biological invasion; propagule pressure can be reduced by inspection and sanitation of the pathway vector (e.g., ships, trucks, humans) at the point(s) of departure and at the point of entry. I demonstrate how the risk analysis framework can be used to more efficiently target incoming ships for inspection and propagule pressure reduction.

A comparison of the adaptations of strains of Lymantria dispar multiple nucleopolyhedrovirus to hosts from spatially isolated populations

The adaptation of pathogens to either their hosts or to environmental conditions is the focus of many current ecological studies. In this work we compared the ability of six spatially-distant Lymantria dispar (gypsy moth) multiple nucleopolyhedrovirus (LdMNPV) strains (three from eastern North America and three from central Asia) to induce acute infection in gypsy moth larvae. We also sequenced the complete genome of one Asian (LdMNPV-27/0) and one North American (LdMNPV-45/0) strain which were used for bioassay. We found that all of the North American virus strains, with the exception of one, demonstrated higher potency than the Asian virus strains, either in North American (Lymantria dispar) larvae or, in Asian (Lymantria dispar asiatica) larvae. Complete genome sequencing revealed two gene deletions in the LdMNPV-27/0 strain: the virus enhancin factor gene (vef-1) and the baculovirus repeated orf gene (bro-p). These deletions were not seen in the LdMNPV-45/0 strain nor in other American strains available in archiving systems. We also found deletions of the bro-e and bro-o genes in LdMNPV-45/0 strain but not in the LdMNPV-27/0 strain. The phylogenetic inference with an alignment of the 37 core gene nucleotide sequences revealed the close relationship of the LdMNPV-45/0 strain with other American strains accessed in GenBank (Ab-a624 and 5-6) while the LdMNPV-27/0 strain was clustered together with the LdMNPV-3054 strain (isolated in Spain) instead of predicted clustering with LdMNPV- 3029 (isolated in Asia). Our study demonstrated that first, different LdMNPV isolates from the same metapopulations of L. dispar exhibit little or no difference in the degree of virulence towards host larvae and second, that locality of host population is not an important driver of LdMNPV virulence. Virulence of LdMNPV is determined only by viral genetics. The genetic differences between North American and Central Asian virus strains are discussed.

Landscape-Level Patterns of Elevated FS1 Asian Allele Frequencies in Populations of Gypsy Moth (Lepidoptera: Erebidae) at a Northern U.S. Boundary.

From a regulatory perspective, Asian gypsy moth is a species complex consisting of three species of Lymantria and two subspecies of Lymantria dispar (L.), differing from the European subspecies, L. dispar dispar (L.), by having consistently flight-capable females. As such, the invasion potential in North America is thought to exceed that of European gypsy moth. USDA-APHIS therefore has a monitoring program to detect Asian gypsy moth at high-risk introduction pathways. Molecular markers are used to improve the diagnosis of Asian gypsy moth. One such marker, which targets the FS1 locus, detects an allele, FS1-A, prevalent in Asian populations but occurring at low frequencies (3-6%) throughout the European gypsy moth's range in North America. However, some locales, such as Minnesota, exhibit elevated FS1-A frequencies. We studied the distribution of the FS1-A allele in northern Minnesota, 2013-2014, assessing spatial patterns in the distribution of the FS1-A allele using Moran's I and using spatial regression techniques to examine if the FS1-A allele was associated with putative movement pathways. We also used time series analysis to discern if temporal patterns in FS1-A or possible introduction events occurred. Our results indicated that FS1-A occurred randomly in space and time. We found no evidence that elevated FS1-A frequencies were associated with movement pathways or possible immigration events into this region over the two years. Elevated frequencies of the FS1-A allele within this region could be due to genetic drift and allelic surfing along the expanding population front, or to selection of physiological or behavioral traits.

The influence of geographic population, age, and mating status on the flight activity of the Asian gypsy moth Lymantria dispar (Lepidoptera: Erebidae) in China

The gypsy moth Lymantria dispar (Linnaeus) is a highly polyphagous herbivore that defoliates a wide variety of tree species. The influence of age and mating status on the flight activity of females was assessed in seven geographically isolated populations from China. We examined flight activity of tethered females using a computer-linked flight mill in the laboratory and found that age significantly influenced flight parameters, including total flight distance, time, and speed. Flight distance peaked in the youngest females (at 1 day of age) and declined with age. No significant differences in flight parameters were found between unmated and mated females. Females from Guizhou showed the strongest flight activity among all seven populations. The total flight distances, flight times, and maximum flight speeds at 1 day of age were 7.50 ± 2.28 km, 2.17 ± 0.51 h, and 6.18 ± 0.51 km/h, respectively. These results provide valuable information regarding the scale and speed of dispersal in hotspots and can improve the management and control of forest damage.

Sequencing and de novo assembly of the Asian gypsy moth transcriptome using the Illumina platform.

The Asian gypsy moth (Lymantria dispar) is a serious pest of forest and shade trees in many Asian and some European countries. However, there have been few studies of L. dispar genetic information and comprehensive genetic analyses of this species are needed in order to understand its genetic and metabolic sensitivities, such as the molting mechanism during larval development. In this study, high-throughput sequencing technology was used to sequence the transcriptome of the Asian subspecies of the gyspy moth, after which a comprehensive analysis of chitin metabolism was undertaken. We generated 37,750,380 high-quality reads and assembled them into contigs. A total of 37,098 unigenes were identified, of which 15,901 were annotated in the NCBI non-redundant protein database and 9,613 were annotated in the Swiss-Prot database. We mapped 4,329 unigenes onto 317 pathways using the Kyoto Encyclopedia of Genes and Genomes Pathway database. Chitin metabolism unigenes were found in the transcriptome and the data indicated that a variety of enzymes was involved in chitin catabolic and biosynthetic pathways.

A Multi-Species TaqMan PCR Assay for the Identification of Asian Gypsy Moths (Lymantria spp.) and Other Invasive Lymantriines of Biosecurity Concern to North America.

Preventing the introduction and establishment of forest invasive alien species (FIAS) such as the Asian gypsy moth (AGM) is a high-priority goal for countries with extensive forest resources such as Canada. The name AGM designates a group of closely related Lymantria species (Lepidoptera: Erebidae: Lymantriinae) comprising two L. dispar subspecies (L. dispar asiatica, L. dispar japonica) and three closely related Lymantria species (L. umbrosa, L. albescens, L. postalba), all considered potential FIAS in North America. Ships entering Canadian ports are inspected for the presence of suspicious gypsy moth eggs, but those of AGM are impossible to distinguish from eggs of innocuous Lymantria species. To assist regulatory agencies in their identification of these insects, we designed a suite of TaqMan® assays that provide significant improvements over existing molecular assays targeting AGM. The assays presented here can identify all three L. dispar subspecies (including the European gypsy moth, L. dispar dispar), the three other Lymantria species comprising the AGM complex, plus five additional Lymantria species that pose a threat to forests in North America. The suite of assays is built as a “molecular key” (analogous to a taxonomic key) and involves several parallel singleplex and multiplex qPCR reactions. Each reaction uses a combination of primers and probes designed to separate taxa through discriminatory annealing. The success of these assays is based on the presence of single nucleotide polymorphisms (SNPs) in the 5’ region of mitochondrial cytochrome c oxidase I (COI) or in its longer, 3’ region, as well as on the presence of an indel in the “FS1” nuclear marker, generating North American and Asian alleles, used here to assess Asian introgression into L. dispar dispar. These assays have the advantage of providing rapid and accurate identification of ten Lymantria species and subspecies considered potential FIAS.

Geographic isolates of Lymantria dispar multiple nucleopolyhedrovirus: Genome sequence analysis and pathogenicity against European and Asian gypsy moth strains

Isolates of the baculovirus species Lymantria dispar multiple nucleopolyhedrovirus have been formulated and applied to suppress outbreaks of the gypsy moth, L. dispar. To evaluate the genetic diversity in this species at the genomic level, the genomes of three isolates from Massachusetts, USA (LdMNPV-Ab-a624), Spain (LdMNPV-3054), and Japan (LdMNPV-3041) were sequenced and compared with four previously determined LdMNPV genome sequences. The LdMNPV genome sequences were collinear and contained the same homologous repeats (hrs) and clusters of baculovirus repeat orf (bro) gene family members in the same relative positions in their genomes, although sequence identities in these regions were low. Of 146 non-bro ORFs annotated in the genome of the representative isolate LdMNPV 5-6, 135 ORFs were found in every other LdMNPV genome, including the 37 core genes of Baculoviridae and other genes conserved in genus Alphabaculovirus. Phylogenetic inference with an alignment of the core gene nucleotide sequences grouped isolates 3041 (Japan) and 2161 (Korea) separately from a cluster containing isolates from Europe, North America, and Russia. To examine phenotypic diversity, bioassays were carried out with a selection of isolates against neonate larvae from three European gypsy moth (Lymantria dispar dispar) and three Asian gypsy moth (Lymantria dispar asiatica and Lymantria dispar japonica) colonies. LdMNPV isolates 2161 (Korea), 3029 (Russia), and 3041 (Japan) exhibited a greater degree of pathogenicity against all L. dispar strains than LdMNPV from a sample of Gypchek. This study provides additional information on the genetic diversity of LdMNPV isolates and their activity against the Asian gypsy moth, a potential invasive pest of North American trees and forests.

Evaluation of the Effects of Light Intensity and Time Interval After the Start of Scotophase on the Female Flight Propensity of Asian Gypsy Moth (Lepidoptera: Erebidae).

Asian gypsy moth, Lymantria dispar L. (Lepidoptera: Erebidae), females are capable of flight, but little is known about what causes the variation in flight propensity that has been observed. The female flight propensity and capability of Asian gypsy moth from seven geographic populations (three from China, two from Russia, one from Japan, and one from Korea) were compared under all combinations of three light intensities (0.05, 0.10, and 0.40 lux) and during three time intervals after the start of scotophase. A total of 567 females were flight tested. Female flight propensity, time to initiate walking, fanning, and flying, and duration of fanning differed significantly among geographic populations. Females were less likely to voluntarily fly during the 0-1-h time interval after the start of scotophase than during the later time intervals (1-2 and 2-3 h), suggesting that the light intensity cue has to occur at the correct time after the expected start of scotophase for flight initiation. Light intensity did not significantly affect the proportion of females that voluntarily flew, but did impact the timing of the walking and fanning preflight behaviors. The interaction between light intensity and time interval after the start of scotophase had a significant effect on the proportion of females that fanned. The proportion of females with sustained flight capability varied among the populations evaluated. These results may aid in determining the risk of Asian gypsy moth dispersal, but further work is needed to assess other factors that play a role in flight propensity.

PCR-RFLP analysis of the ITS2 region to identify Japanese Lymantria species (Lepidoptera: Lymantriidae)

In this study, a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)-based method for species identification was applied to eight Japanese Lymantria species, including the four “Asian gypsy moth” species [Lymantria dispar japonica (Motschulsky), Lymantria umbrosa (Butler), Lymantria albescens Hori and Umeno, and Lymantria postalba Inoue] and four other species [Lymantria xylina Swinhoe, Lymantria mathura Moore, Lymantria monacha (Linnaeus), and Lymantria lucescens (Butler)]. We sequenced the second internal transcribed spacer (ITS2) region of nuclear ribosomal DNA (rDNA) of one individual from each of the eight species. We analyzed the recognition sites of a restriction endonuclease (MnlI) to identify the Lymantria species by PCR-RFLP. We then applied PCR-RFLP analysis to 330 individuals from 54 populations of the 8 species. As a result, these species could be correctly identified with the exception of two species pairs (L. dispar japonica and L. umbrosa; L. albescens and L. postalba). These results suggest that this PCR-RFLP method based on the ITS2 region is useful for identifying Japanese Lymantria species.