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Changes in grain trading between Russian Far Eastern ports and North America in the late 1980s and early 1990s led to introductions of Lymantria dispar asiatica (Vnukovskij), formerly called the Asian gypsy moth (AGM), and the recognition of a new pathway for its transport. Unlike the pathways commonly used and regulated for commodities and for packaging material containing pest organisms, this presented a unique pathway for AGM. Vessels departing from the Russia Far East transiting to ports on the west coast of the United States (US) and Canada (CA) were infested with life stages of AGM and related species. Upon arrival in North America, eggs oviposited on the surfaces of the ships and its cargo hatched with larvae ballooning to vegetation surrounding port areas leading to the potential establishment of AGM populations. A multi layered monitoring and inspection program was developed to mitigate this risk of introduction, initially for Russian Far Eastern ports, and eventually to include specified areas of other Asian countries. In this article, we summarize and review the integral parts of this mitigation program, which include: risk assessments, AGM monitoring in foreign and domestic ports, vessel cleaning and certification by a government Plant Protection Organization (PPO) or its proxy, AGM surveillance, and eradication of introduced AGM in ports and other areas within the United States and Canada. The uniqueness of this program is characterized by its complexity, which involves coordinated efforts of PPOs, the use of various inspection organizations, and the support of ocean transportation industries. [ABSTRACT FROM AUTHOR]

Sex differences in gene expression have been extensively documented, but little is known about these differences in parasitoid species that are widely applied to control pests. Brachymeria lasus is a solitary parasitoid species and has been evaluated as a potential candidate for release to control Lymantria dispar. In this study, gender differences in B. lasus were investigated using Illumina-based transcriptomic analysis. The resulting 37,453 unigene annotations provided a large amount of useful data for molecular studies of B. lasus. A total of 1416 differentially expressed genes were identified between females and males, and the majority of the sex-biased genes were female biased. Gene Ontology (GO) and Pathway enrichment analyses showed that (1) the functional categories DNA replication, fatty acid biosynthesis, and metabolism were enhanced in females and that (2) the only pathway enriched in males was phototransduction, while the GO subcategories enriched in males were those involved in membrane and ion transport. In addition, thirteen genes involving transient receptor potential (TRP) channels were annotated in B. lasus. We further explored and discussed the functions of TRPs in sensory signaling of light and temperature. In general, this study provides new molecular insights into the biological and sexually dimorphic traits of parasitoids, which may improve the application of these insects to the biological control of pests. [ABSTRACT FROM AUTHOR]

Since its accidental introduction to Massachusetts in the late 1800s, the European gypsy moth (EGM; Lymantria dispar dispar) has become a major defoliator in North American forests. However, in part because females are flightless, the spread of the EGM across the United States and Canada has been relatively slow over the past 150 years. In contrast, females of the Asian gypsy moth (AGM; Lymantria dispar asiatica) subspecies have fully developed wings and can fly, thereby posing a serious economic threat if populations are established in North America. To explore the genetic determinants of these phenotypic differences, we sequenced and annotated a draft genome of L. dispar and used it to identify genetic variation between EGM and AGM populations. The 865-Mb gypsy moth genome is the largest Lepidoptera genome sequenced to date and encodes ~13,300 proteins. Gene ontology analyses of EGM and AGM samples revealed divergence between these populations in genes enriched for several gene ontology categories related to muscle adaptation, chemosensory communication, detoxification of food plant foliage, and immunity. These genetic differences likely contribute to variations in flight ability, chemical sensing, and pathogen interactions among EGM and AGM populations. Finally, we use our new genomic and transcriptomic tools to provide insights into genome-wide gene-expression changes of the gypsy moth after viral infection. Characterizing the immunological response of gypsy moths to virus infection may aid in the improvement of virus-based bioinsecticides currently used to control larval populations. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Abstract: A morphofunctional investigation of the different neuronal subpopulations projecting through each of the nerves IV–VI emerging bilaterally from the terminal abdominal ganglion (TAG) was correlated with the octopaminergic activity in the ganglion that controls the ovipositor movements associated with calling behavior in the female gypsy moth Lymantria dispar. Tetramethylrodamine‐dextran backfills from nerve stumps resulted in a relatively low number of TAG projections, ranging from 12 to 13 for nerve pair IV, 12 to 14 for nerve pair V, and 8 to 9 for nerve pair VI. Furthermore, as assessed by electrophysiological recordings, a number of fibers within each of these nerves displays spontaneous tonic activity, also when the ganglion is fully disconnected from the ventral nerve cord (VNC). Octopamine (OA) applications to the TAG strongly enhanced the activity of these nerves, either by increasing the firing rate of a number of spontaneously firing units or by recruiting new ones. This octopaminergic activity affected calling behavior, and specifically the muscle activity leading to cycling extensions of the intersegmental membrane (IM) between segments VIII and IX (ovipositor). Our results indicate that in the female gypsy moth the octopaminergic neural activity of the TAG is coupled with extensions and retractions of IM for the purpose of releasing pheromone, where motor units innervated by nerve pair IV appear antagonistic with respect to those innervated by nerve pair V. [ABSTRACT FROM AUTHOR]

1. It has been predicted that temperature increases of 3.6–5.8 °C would shift the northern distribution limit of Lymantria monacha (Linnaeus) and Lymantria dispar (Linnaeus) by 500–700 km, but these predictions ignore the effects of minimum winter temperatures. It was hypothesised that winter cold can limit range expansion due to high egg mortality in cold temperatures. 2. The present study determined the supercooling points of overwintering eggs of these forest pests, and compared these with recent minimum winter temperatures in the areas of origin of three populations. Eggs from one L. monacha and one L. dispar population from the species' core distribution area in Germany were included, as well as L. monacha eggs from Finland, near the northern border of the species' distribution. 3. The median supercooling points of both species were more than 10 °C lower than the median minimum winter temperatures of their areas of origin, and the median supercooling points of Finnish and German L. monacha eggs did not differ significantly. The median supercooling point of German L. monacha eggs differed from that of German L. dispar eggs. 4. Previous literature on the topic is referenced, and translations of the old German and Russian sources are given. Based on these results, it is argued that the frequent claim that L. monacha eggs can survive cold down to −40 °C is unsupported, with a value near −30 °C being a more likely limit. 5. Winter cold alone can limit the predicted range shifts of these species to 200–300 km under 3.6–5.8 °C increase scenarios, which is less than half the value of earlier estimates. [ABSTRACT FROM AUTHOR]

Camptothecin (CPT), a monoterpene indole alkaloid, is a potent inhibitor of eukaryotic topoisomerase I (Top 1). Because of this property, several derivatives of CPT are widely used as chemotherapeutic agents. The compound is produced by several plant species, including Nothapodytes nimmoniana (Family: Icacinaceae) presumably as a deterrent to insect pests. Here, we report, a lepidopteran larva, Lymantria sp. of Lymantriidae family which feeds voraciously on the leaves of N. nimmoniana, without any adverse consequences. Larval body weight and molting period were unaffected despite captive feeding of the larva with CPT enriched leaves. Mass spectrometric analysis indicated that nearly 46% of the ingested CPT was excreted while the rest was sequestered predominantly in the exuviae and setae (~35%). Although most of the CPT was in the parental form as found in the plant, traces of inactive, sulfated forms of CPT were recovered from the larva. Compared to that in plant, there were no critical mutations at the CPT binding domain of the insect’s Top 1. The gut pH of the larva was alkaline (pH 10.0). The alkaline gut environment converts CPT from its active, lactone form to inactive, carboxylate form. It is likely that such conversion might help the larva to reduce the overall burden of CPT in its gut. We discuss the results in the context of the mechanisms of resistance adapted by insects to plant toxins. [ABSTRACT FROM AUTHOR]

An accurate quantitative relationship between key characteristics of an insect population, such as season-long and peak abundances, can be very useful in pest management programs. To the best of our knowledge, no such relationship has yet been established. Here we establish a predictive linear relationship between insect catch Mpw during the week of peak abundance, the length of seasonal flight period, F (number of weeks) and season-long cumulative catch (abundance) A = 0.41MpwF. The derivation of the equation is based on several general assumptions and does not involve fitting to experimental data, which implies generality of the result. A quantitative criterion for the validity of the model is presented. The equation was tested using extensive data collected on captures of male gypsy moths Lymantria dispar (L.) (Lepidoptera: Erebidae) in pheromone-baited traps during 15 years. The model was also tested using trap catch data for two species of mosquitoes, Culex pipiens (L.) (Diptera: Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae), in Gravid and BG-sentinel mosquito traps, respectively. The simple, parameter-free equation approximates experimental data points with relative error of 13% and R2 = 0.997, across all of the species tested. For gypsy moth, we also related season-long and weekly trap catches to the daily trap catches during peak flight. We describe several usage scenarios, in which the derived relationships are employed to help link results of small-scale field studies to the operational pest management programs. [ABSTRACT FROM AUTHOR]

RNA interference (RNAi) technology uses dsRNAs to silence specific targeted genes by downregulating their expression. It has become a potent tool for functional and regulatory studies of insect genes and has potential to be applied for insect control. Though it has been challenging to generate effective RNAi in lepidopteran insects, in the current study this technology was applied to develop specific RNAi-based molecular tools that could be used to negatively impact the invasive lepidopteran forest pest, gypsy moth (GM). GM midgut-specific genes were selected for dsRNA design from larval transcriptome profiles. Two methods were used to produce specific dsRNAs, bacterial expression and in vitro synthesis, which were then fed per os to GM larvae. Depletion of uncharacterized gene targets known as locus 365 and locus 28365, or their stacked combination, depleted target transcripts in a sequence specific manner and resulted in 60% reduction in body mass. Treated GM females that were able to moult to the adult stage displayed an approximately two-fold reduction in egg masses. These have potential to be developed as molecular biopesticides for GM. [ABSTRACT FROM AUTHOR]

Bioaccumulation and excretion of heavy metals in insects is only partially clarified. We have investigated cadmium accumulation in the feces, head and integument of Lymantria dispar (Lepidoptera: Lymantriidae) larvae exposed to chronic dietary intake of cadmium. The aim of the experiment was to establish modalities of metal accumulation, primarily cadmium, as well as changes in fitness traits in two insect populations receiving 50 or 100 μg Cd/g dry food. The egg-masses originated from two localities: a protected nature reserve (unpolluted population) and an area near a busy highway (polluted population) in Serbia. At both added dietary levels cadmium concentration was highest in feces. Small alterations in metal concentrations after both cadmium treatments were detected in the integument. It was established that irrespective of population origin, the modality of decline of larval mass and relative growth rate (RGR) was similar. Concentrations of Cu and Zn in the integument were approximately the same regardless of dietary cadmium intake. However, cadmium accumulation in the examined tissues, as well as variability of fitness traits depended on population origin and cadmium concentration. In larvae from both populations not given contaminated food the head was the organ with the greatest accumulation of trace elements. Our results reveal how invasive phytophagous insects cope with high metal concentrations in their food by body mass (RGR) reduction and energy allocation towards processes that enable accumulation of cadmium and other trace elements in different tissues. [ABSTRACT FROM AUTHOR]

Host innate immunity is one of the factors that determines the resistance of insects to their entomopathogens. In the research reported here we studied whether or not phenoloxidase (PO), a key enzyme in the melanogenesis component of humoral immunity of insects, plays a role in the protection of Lymantria dispar larvae from infection by L. dispar multiple nucleopolyhedrovirus. We studied two types of viral infection: overt and covert. The following lines of investigation were tested: i) the intravital individual estimation of baseline PO activity in haemolymph plasma followed by virus challenging; ii) the specific inhibition of PO activity in vivo by peroral treatment of infected larvae with phenylthiourea (PTU), a competitive inhibitor of PO; iii) the evaluation of PO activity in the haemolymph plasma after larval starvation. Starvation is a stress that activates the covert infection to an overt form. All of these experiments did not show a relationship between PO activity in haemolymph plasma of L. dispar larvae and larval susceptibility to baculovirus. Moreover, starvation-induced activation of covert viral infection to an overt form occurred in 70 percent of virus-carrying larvae against the background of a dramatic increase of PO activity in haemolymph plasma in the insects studied. Our conclusion is that in L. dispar larvae PO activity is not a predictor of host resistance to baculovirus. [ABSTRACT FROM AUTHOR]

The nun moth, Lymantria monacha L., is one of the most important defoliators of Eurasian coniferous forests. Outbreaks during 2011-2015 in the natural/planted larch, and larch-birch mixed forests of the Greater Khingan Range in Inner Mongolia, China, caused tremendous timber losses from severe defoliation and tree mortality. A series of trapping experiments were conducted in these outbreak areas to evaluate the efficacy of a synthetic species-specific pheromone lure based on the female pheromone blend of European nun moth populations. Our results clearly show that the nun moth in Inner Mongolia is highly and specifically attracted to this synthetic pheromone, with few gypsy moths ( Lymantria dispar) captured. Flight activity monitoring of L. monacha male moths using pheromone-baited Unitraps at 2 locations during the summer of 2015 indicated that the flight period started in mid-July, peaking in early August at both locations. Based on male moth captures, there was a strong diurnal rhythm of flight activity throughout the entire scotophase, peaking between 22:00 and 24:00. Unitraps and wing traps had significantly and surprisingly higher catches than the gypsy moth traps. Unitraps fastened to tree trunks 2 m above ground caught significantly more male moths than those at the ground level or at 5 m height. Male L. monacha moths can be attracted to pheromone-baited traps in open areas 150-200 m distant from the infested forest edge. Our data should allow improvement on the performance of pheromone-baited traps for monitoring or mass-trapping to combat outbreaks of this pest in northeastern China. [ABSTRACT FROM AUTHOR]

This study investigated the statistical relationship between defoliation in pine forests infested by nun moths (Lymantria monacha) and the spectral bands of the RapidEye sensor, including the derived normalized difference vegetation index (NDVI) and the normalized difference red-edge index (NDRE). The strength of the relationship between the spectral variables and the ground reference samples of percent remaining foliage (PRF) was assessed over three test years by the Spearman's ρ correlation coefficient, revealing the following ranking order (from high to low ρ): NDRE, NDVI, red, NIR, green, blue, and rededge. A special focus was directed at the vegetation indices. In both discriminant analyses and decision tree classification, the NDRE yielded higher classification accuracy in the defoliation classes containing none to moderate levels of defoliation, whereas the NDVI yielded higher classification accuracy in the defoliation classes representing severe or complete defoliation. We concluded that the NDRE and the NDVI respond very similarly to changes in the amount of foliage, but exhibit particular strengths at different defoliation levels. Combining the NDRE and the NDVI in one discriminant function, the average gain of overall accuracy amounted to 7.8 percentage points compared to the NDRE only, and 7.4 percentage points compared to the NDVI only. Using both vegetation indices in a machine-learning-based decision tree classifier, the overall accuracy further improved and reached 81% for the test year 2012, 71% for 2013, and 79% for the test year 2014. [ABSTRACT FROM AUTHOR]

According to the present research trend and legislative framework of the major world regions fostering the development and use of low-risk biocidal and plant protection products for Integrated Pest Management, the discovery of new entomopathogenic bacterial species and isolates bearing insecticidal traits against novel targets is needed in the near future. In this study, the oral insecticidal activity of bacteria isolated from septicaemicGalleria mellonellalarvae was assessed against different insect pests of agricultural, forest and medical-veterinary importance, including Lepidoptera (Lymantria disparandMalacosoma neustria) and Diptera (Musca domesticaandCeratitis capitata). This led to the identification of new entomopathogenic bacterial isolates belonging to different species includingAlcaligenes aquatilis, Alcaligenes faecalis, Enterococcus mundtii, Pseudomonas protegens, Serratia nematodiphila, Serratia marcescensandStenotrophomonas maltophilia. The majority of these isolates were toxic to gypsy mothL. disparand, more moderately, to lackey mothM. neustrialarvae. Variable degrees of virulence were observed on fly species. While the insecticidal action of some of these species was previously observed, we report for the first time the insecticidal properties of specific strains against a wider target range. [ABSTRACT FROM PUBLISHER]

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 highrisk 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. [ABSTRACT FROM AUTHOR]

When pheromone traps are used for detection of an invasive pest and then delimitation of its distribution, an unresolved issue is the interpretation of failure to capture any target insects. Is a population present but not detected, a so-called false negative? Using the gypsy moth ( Lymantria dispar) as an exemplar, we modeled the probability of males being captured in traps deployed at densities typical for surveillance (1 per 2.6 km or 1 per mi) and delimitation (up to 49 per 2.6 km). The simulations used a dynamic wind model generating a turbulent plume structure and varying wind direction, and a behavior model based on the documented maneuvers of gypsy moths during plume acquisition and along-plume navigation. Several strategies of plume acquisition using Correlated Random Walks were compared to ensure that the generated dispersions over three days were not either overly clumped or ranged many km. Virtual moths were released into virtual space with patterns mimicking prior releases of gypsy moth males in Massachusetts at varying distance from a baited trap. In general, capture rates of virtual and real moths at varying trap densities were similar. One application of this approach was to estimate through bootstrapping the probabilities of not detecting populations having densities ranging from 1 to 100 moths per 2.6 km and using traps that varied from 25 to 100 % in their efficiencies of capture. Low-level populations (e.g., 20-30 per 2.6 km) often were not detected with one trap per 2.6 km, especially when traps had low efficiencies. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

A high prevalence of nuclear polyhedrosis has been observed in larval outbreaks of Lymantria mathura in Iwate Prefecture, Japan. However, the virus responsible has not been identified. Here the virus was designated Lymantria mathura nucleopolyhedrovirus (LymaNPV), based on partial sequence results of the lef-8 , lef-9 , and polh genes and transmission electron microscopic observations. Diagnosis by polymerase chain reaction targeting of the partial polh gene and a subsequent restriction fragment length polymorphism (RFLP) analysis indicated that LymaNPV was an exclusive causative agent for the nuclear polyhedrosis of the L . mathura larvae. LymaNPV was also detected from a very small fraction of L. dispar larvae that co-occurred with L. mathura larvae where the prevalence of LymaNPV-caused disease was high. A bioassay using LymaNPV against L. dispar larvae produced infection with the inoculated virus. This finding was consistent with the results of the field sample analysis. LymaNPV contained six genome types in the L. mathura populations sampled, as determined by RFLP of LymaNPV genomic DNA and southern blot hybridization analyses. None of the genome types was unique to any sampling site, indicating that some flow had occurred among sites. However, genome-type composition seemed to differ among sites. This study provides basic information about the interaction between L. mathura and LymaNPV. [ABSTRACT FROM AUTHOR]

Diverse parasite taxa share hosts both at the population level and within individual hosts, and their interactions, ranging from competitive exclusion to facilitation, can drive community structure and dynamics. Emergent pathogens have the potential to greatly alter community interactions. We found that an emergent fungal entomopathogen dominated pre-existing lethal parasites in populations of the forest defoliating gypsy moth, Lymantria dispar. The parasite community was composed of the fungus and four parasitoid species that only develop successfully after they kill the host, and a virus that produces viable propagules before the host has died. A low-density site was sampled over 17 years and compared with 66 sites across a range of host densities, including outbreaks. The emergent fungal pathogen and competing parasitoids rarely co-infected host individuals because each taxa must kill its host. The virus was not present at low host densities, but successfully co-infected with all other parasite species. In fact, there was facilitation between the virus and one parasitoid species hosting a polydnavirus. This newly formed parasite community, altered by an emergent pathogen, is shaped both by parasite response to host density and relative abilities of parasites to co-inhabit the same host individuals. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Liparis aphylla G.A. Romero & Garay was previously known only from two herbarium specimens collected in 1945 and 1977 in Ecuador and Colombia, respectively. This little-known species is hereby reported for the first time for Peru. An updated description, line illustration, color photographs and distribution map of L. aphylla, as well as an identification key to the Peruvian species of Liparis are provided. [ABSTRACT FROM AUTHOR]

The Indian gypsy moth ( Lymantria obfuscata ) is a pest of national significance, which attacks a wide range of forest and fruit crops in India. The outbreaks of this pest over the past few years spurred interest towards sustainable biointensive approaches for quality fruit production of apple and walnut for agricultural export in Jammu & Kashmir (J & K). The exploratory survey's were conducted for the isolation of native strains of L. obfuscata multiple nucleopolyhedrovirus (LyobMNPV) and field evaluation was conducted to determine the most promising strain as a potential biological control agent of this pest. The virus was recovered from different locations of J&K with its natural incidence varying from 0.77 to 7.43 percent. This is the first report from the Indian sub-continent and worldwide of the occurrence of NPVs in natural larval populations of L. obfuscata . The in vivo mass production was optimised by inoculating per os late 3rd instar L. obfuscata larvae with an inoculum dose of 1.44 × 10 5 OBs/larva and followed by a 10 day incubation period. To prevent the disintegration of larval cuticle and microbial contamination, moribund larvae were harvested with an average yield of 1.42 × 10 8 OBs/larva. The field application of LyobMNPV against the host populations on apple and willow with the pre-standardised dosage of 2.5 × 10 12 OBs/ha reduced the larval population density by 25–63%. In the year following application, the larval population density in the treated blocks was 19–28% lower as compared to controls on both apple and willow. This suggests that the virus has considerable potential as a biocontrol agent and the timely application with LyobMNPV on non-fruit trees such as willow which are the preferred hosts for egg laying may help to initiate viral epizootics to keep population outbreaks under check. [ABSTRACT FROM AUTHOR]

A taxonomic review of the Korean L ymantria Hübner, 1819 was conducted. A total of nine species of five subgenera with two unrecorded species are listed: L ymantria ( P orthetria) dispar Linnaeus 1758, L. ( P.) xylina Swinhoe 1903, L. ( L ymantria) monacha ( Linnaeus 1758), L. ( L.) minomonis Matsumura 1933 (new to Korea), L. ( L.) similis monachoides Schintlimeister 2004 (new to Korea), L. ( L.) lucescens ( Butler 1881), L. ( N yctria) mathura Moore 1865, L. ( C ollentria) fumida Butler 1877, and L. ( S pinotria) bantaizana Matsumura 1933. L ymantria ( L ymantria) minomonis and L. ( L.) similis monachoides are newly added to the Korean fauna. L ymantria ( L.) minomonis was found only on Bogildo Island of Jeollanam-do in the southern part of Korea, and L. ( L.) similis monachoides was collected in central Korea. L ymantria ( P orthetria) xylina and L. ( C ollentria) fumida were not examined in this study, and it is considered that the previous records were due to misidentification or they are only distributed in the northern part of the Korean Peninsula. We provide diagnoses of two unrecorded species and adult habitus and genitalia photos of the Korean L ymantria species. [ABSTRACT FROM AUTHOR]

1. Globally, vast areas of forest are currently threatened by Lymantria dispar L. and Phytophthora species, which cause widespread declines and cascading ecological impacts. One important aim of evolutionary and ecological studies is to understand their interactions. 2. The present study tests whether Quercus rubra L. trees naturally infected with P. plurivora T. Jung & T.I. Burgess or free of infection are more suitable for L. dispar herbivory, and if relationships between L. dispar performance and herbivory may vary depending on whether trees are infected or free of infection. 3. In choice tests, the consumed area of leaves from trees infected by P. plurivora was four times larger than that from non-infected trees, probably because the increased values of N, soluble protein, and water content observed in the leaves of infected trees enhanced acceptability. Although larval performance was better in Phytophthora-infected trees, relationships between larval performance and defoliation did not significantly interact with the health status of trees. 4. The present results suggest that the impact of P. plurivora on natural and managed ecosystems may generate a positive feedback loop for oak decline. The link between the behavioural and physiological responses of L. dispar to infected trees and the population growth in nature deserves further investigation. [ABSTRACT FROM AUTHOR]

DNA barcoding and morphological analyses of Korean Lymantria (Erebidae, Lepidoptera) were conducted for quarantine inspection. In DNA barcoding, Lymantria dispar identified through quarantine inspection was distinguished as three species, L. dispar asiatica, L. albescens, and L. xylina. Lymantria monacha, which is known as a single species in Korea, is revealed as containing three species, L. monacha, L. minomonis, and L. sugii. At the subspecies level, L. dispar dispar formed a single cluster, whereas L. d. asiatica and L. d. japonica formed a cluster containing both subspecies. In morphological re-examination on DNA barcoding results, L. dispar was distinguished from L. albescens by wing pattern, and from L. xylina by papillae anale. L. monacha and the related species were hard to be distinct from each other by using wing pattern, but it was easily distinct through comparison of genitalia. Therefore, DNA barcoding led to accurate identification in species level, but in subspecies level, only a taxon showing geographically far distance was discriminated from the others. These results may provide a taxonomic outline of the Korean Lymantria fauna and may be used as an identification reference for Lymantria species during quarantine inspection. [ABSTRACT FROM AUTHOR]

We investigated the effects of acute and chronic treatments with cadmium at 10 μg Cd/g dry food and 30 μg Cd/g dry food on α-glucosidase activity of the 4th instar larvae ofLymantria dispar(Lepidoptera: Lymantriidae) as well as subsequent recovery. Enzyme inhibition was recorded during acute exposure to 30 μg Cd/g dry food and during chronic treatment at the lower metal concentration. After three days recovery from 10 μg Cd/g dry food, the α-glucosidase activity returned to the control level. One-way ANOVA showed that cadmium significantly influenced the activity of α-glucosidase during all treatments. The index of phenotypic plasticity was higher during chronic treatment at 10 μg Cd/g dry food than at 30 μg Cd/g as well as during the recovery. We detected four glucosidase isoforms by NATIVE PAGE. The activities and expressions of the isoforms depended on both larval genotype and cadmium treatment. [ABSTRACT FROM AUTHOR]

Tympanal ears of female gypsy moths Lymantria dispar dispar ( L.) ( Lepidoptera: Erebidae: Lymantriinae) are reportedly more sensitive than ears of conspecific males to sounds below 20 kHz. The hypothesis is tested that this differential sensitivity is a result of sex-specific functional roles of sound during sexual communication, with males sending and females receiving acoustic signals. Analyses of sounds produced by flying males reveal a 33-Hz wing beat frequency and 14-kHz associated clicks, which remain unchanged in the presence of female sex pheromone. Females exposed to playback sounds of flying conspecific males respond with wing raising, fluttering and walking, generating distinctive visual signals that may be utilized by mate-seeking males at close range. By contrast, females exposed to playback sounds of flying heterospecific males ( Lymantria fumida Butler) do not exhibit the above behavioural responses. Laser Doppler vibrometry reveals that female tympana are particularly sensitive to frequencies in the range produced by flying conspecific males, including the 33-Hz wing beat frequency, as well as the 7-kHz fundamental frequency and 14-kHz dominant frequency of associated clicks. These results support the hypothesis that the female L. dispar ear is tuned to sounds of flying conspecific males. Based on previous findings and the data of the present study, sexual communication in L. dispar appears to proceed as: (i) females emitting sex pheromone that attracts males; (ii) males flying toward calling females; and (iii) sound signals from flying males at close range inducing movement in females, which, in turn, provides visual signals that could orient males toward females. [ABSTRACT FROM AUTHOR]

The gypsy moth ( Lymantria dispar) is an insect folivore that feeds on a broad range of hosts, and undergoes intermittent outbreaks that cause extensive tree mortality. Like many other herbivorous insects, gypsy moth larvae consume a substrate that is low in nitrogen. Gypsy moth larvae have been known to cannibalize under crowded conditions in the laboratory. In this study, we assessed the influence of nitrogen and density on cannibalism behavior in gypsy moth larvae. Cannibalism rates increased with decreased nitrogen and increased density. There was no interaction between these two parameters. Developmental experiments confirmed that low dietary nitrogen is detrimental, in agreement with previous studies. In a second experiment, we assessed the influence of previous cannibalism experiences on subsequent cannibalism behavior. Gypsy moth larvae that had previously cannibalized other larvae subsequently exhibited higher cannibalism rates than those larvae that had not cannibalized. In conclusion, low nitrogen, high larval density, and previous cannibalism experience are important factors contributing to gypsy moth larval cannibalism. Future studies are needed to estimate benefits to larvae, and to more closely approximate field conditions. [ABSTRACT FROM AUTHOR]

[Objective] The paper was to explore the correlation between inclusion contents in Casuarina spp. twigs and its resistance to Lymantria xylina. [Method] Three types of representative Casuarina spp. strains including resistant strains(Zhanjiang 3, Hui 76, Hui 83, Guangdong A8-2), moderate resistant strains(Pingtan 2, Hui 1, Long 7-18) and susceptible strains(Dongshan 2, Guangdong 501, Kangfeng) were used as materials to determine the contents of six kinds of inclusions, total phenol, tannin, flavonoids, protein, soluble sugar and polysaccharide. [Result] The resistant strains had higher flavonoids, tannin and total phenol contents and lower soluble sugar and protein contents than the susceptible strains, while the content of polysaccharide acted randomly. The grey correlativity analysis showed that resistance degree, pupation rate and pupal weight had high correlation with protein and soluble sugar contents, with correlation index equal to or above 0.5000. [Conclusion] The study provided a theoretical basis for screening and promotion of resistant strains against L. xylina. [ABSTRACT FROM AUTHOR]

Adult Lymantria dispar japonica (females only) and Lymantria mathura (both sexes) flew to commercial lighting during the night in Takizawa Village, Iwate Prefecture, Japan, in large numbers during the first week of August 2008. Males of L. d. japonica were conspicuously absent while 93.8% of responding females were mated and subsequently laid an average of 419.2 eggs each post-flight. For L. mathura females, only 33.9% were mated during the first half of the night and each mated female carried on average 717.4 eggs (max. 1065). Egg counts were facilitated by a newly developed egg mass matrix digestion process and an approximation of egg count based on egg mass dry weight was calculated. These two moth species have dissimilar behavioral strategies- L. d. japonica is diurnal, mated during the afternoon pre-flight and females flew to the lighting after mating. In contrast, L. mathura is nocturnal and both sexes responded to the lighting; females generally arrived as virgins (unfertilized) to then mate with males as the night progressed. Lack of a moth flight in 2009 illustrated dramatic interannual population fluctuation prevalent among many lymantriine moths. An illustration of a 2013 mass flight in Ono City, Fukui Prefecture, suggests a more frequent occurrence of such flights in both time and space. [ABSTRACT FROM AUTHOR]

The spectral sensitivity of the compound eye in three gypsy moth species from six different geographical regions ( Lymantria dispar asiatica Vnukovskij [Asian gypsy moth], Lymantria dispar japonica Motschulsky [Japanese gypsy moth], and Lymantria dispar dispar L. [North American gypsy moth]) was tested electrophysiologically in the wavelength region 300-700 nm. For all moths examined, a maximum response occurred in the 480-520-nm range (blue-green region) with a shoulder peak occurring at 460 nm. A smaller, secondary peak was observed for both sexes at the 340-380-nm range, which is in the region considered behaviorally maximal in night-flying insects. No peaks in sensitivity were observed between 520 and 700 nm (red region) for any of the moths tested. Based on our retinal recording data, a short wavelength blocking filter with a transition wavelength near 500 nm should reduce gypsy moth attraction to artificial lighting sources. This would help reduce the number of Lymantria-infested ships traveling to and from foreign ports. [ABSTRACT FROM AUTHOR]

Highlights: [•] We evaluated the identity and genetic variation of L. dispar NPV isolates. [•] L. dispar NPVs from around the world are variants of the alphabaculovirus LdMNPV. [•] LdMNPV isolates occur in two groups: European/North American & Asian. [•] LdMNPV-2161 genome sequence is similar to LdMNPV 5–6 with ORF content differences. [•] 3.2-fold range in pathogenicity of North American and Asian isolates in bioassays. [Copyright &y& Elsevier]

A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)-based method for species identification was applied to seven Japanese Lymantria species, including four Asian gypsy moth (AGM) species. We sequenced the partial end of the cytochrome c oxidase I (COI) gene, tRNA leucine, COII gene, and partial end of the tRNA lysine in mitochondrial DNA (mtDNA) for one individual of each of the seven species. We analyzed the recognition sites of three restriction endonucleases and constructed a scheme for Lymantria species identification using PCR-RFLP. We then applied the scheme to 291 individuals from 45 populations of seven species. We found that all seven species were correctly identified using PCR-RFLP. These results suggest that PCR-RFLP is useful for identifying Japanese Lymantria species, which may be detected at Japanese ports. [ABSTRACT FROM AUTHOR]

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The gypsy moth, Lymantria dispar, is a non-native species that continues to invade areas in North America. It spreads generally through stratified dispersal where local growth and diffusive spread are coupled with long-distance jumps ahead of the leading edge. Long-distance jumps due to anthropogenic movement of life stages is a well-documented spread mechanism. Another mechanism is the atmospheric transport of early instars and adult males, believed to occur over short distances. However, empirical gypsy moth population data continue to support the possibility of alternative methods of long-range dispersal. Such dispersal events seemed to have occurred in the mid- to late-1990s with spread across Lake Michigan to Wisconsin. Such dispersal would be against the prevailing wind flow for the area and would have crossed a significant physical barrier (Lake Michigan). The climatology of the region shows that vigorous cyclones can result in strong easterly winds in the area at the time when early instars are present. It is hypothesized that these storms would enable individuals to be blown across the Lake and explain the appearance of new population centers observed at several locations on the western shore of Lake Michigan nearly simultaneously. A synoptic climatology model coupled with population dynamics data from the area was parameterized to show an association between transport events and population spread from 1996 to 2007. This work highlights the importance of atmospheric transport events relative to the invasion dynamics of the gypsy moth, and serves as a model for understanding this mechanism of spread in other related biological invasions. [ABSTRACT FROM AUTHOR]

Mathuralure ( 1) is the major sex pheromone component of the pink gypsy moth Lymantria mathura, a potentially devastating invasive species to North America. To support population monitoring of this moth, a gram-scale synthesis of (-)-mathuralure ( 1) was developed. This process relies on coupling an alkynyl lithium species with a chloroepoxide and provides access to the natural product in a 10% yield over 10 steps. [ABSTRACT FROM AUTHOR]

We tested the effects of defoliation-induced changes in silver birch, Betula pendula, foliar chemistry (delayed induced resistance, DIR) on the fitness and immune defense of the gypsy moth, Lymantria dispar. We measured larval developmental time, pupal weight, rate of survival to the adult stage, and five characteristics of larval immune defense: (1) encapsulation response; (2) phenoloxidase activity; (3) hemocyte concentration and (4) lysozyme-like activity in the hemolymph; and (5) resistance to infection by L. dispar nucleopolyhedrovirus (LdMNPV). The latter is an entomopathogenic baculovirus that often causes epizootics during outbreaks of L. dispar. We also measured the involvement of foliage non-tannin phenolic compounds in resistance of B. pendula to herbivory as well as the relationship between the compounds we identified and L. dispar development, growth, and survival. Leaves of B. pendula with previous defoliation history contained increased levels of myricetin glycoside, two flavonoid aglycones (acacetin and tetrahydroxy-flavone dimethyl ether), as well as one unidentified simple phenolic. The concentrations of two glycosides of quercetin, as well as the content of one unidentified flavonoid glycoside were significantly decreased under defoliation treatment. DIR of B. pendula retarded larval growth rate and increased lysozyme-like activity in the hemolymph, but did not affect encapsulation response, phenoloxidase activity, or hemocyte count. We did not find any DIR-mediated tritrophic interactions among birch, gypsy moth, and LdMNPV. After viral inoculation, the mean hemocyte counts in larvae reared on an individual tree correlated significantly with the survival of larvae reared on that same tree, indicating that hemocyte density in hemolymph might be associated with resistance to viral infection. We found a strong positive correlation between the concentration of 1-(4″-hydroxyphenyl)-3′-oxopropyl-β-D-glucopyranose and L. dispar survival rate, which may indicate an unlikely role of this dominant non-tannin phenolic in B. pendula defense against L. dispar. Our study also shows that several immune characteristics of insects that function as barriers against different groups of parasites are differently affected by plant induced defenses. This underscores the importance of considering multiple factors when characterizing barriers to insect immunity. [ABSTRACT FROM AUTHOR]

Species delimitation directly impacts on global biosecurity. It is a critical element in the decisions made by national governments in regard to the flow of trade and to the biosecurity measures imposed to protect countries from the threat of invasive species. Here we outline a novel approach to species delimitation, "tip to root", for two highly invasive insect pests, Bemisia tabaci (sweetpotato whitefly) and Lymantria dispar (Asian gypsy moth). Both species are of concern to biosecurity, but illustrate the extremes of phylogenetic resolution that present the most complex delimitation issues for biosecurity; B. tabaci having extremely high intraspecific genetic variability and L. dispar composed of relatively indistinct subspecies. This study tests a series of analytical options to determine their applicability as tools to provide more rigorous species delimitation measures and consequently more defensible species assignments and identification of unknowns for biosecurity. Data from established DNA barcode datasets (COI), which are becoming increasingly considered for adoption in biosecurity, were used here as an example. The analytical approaches included the commonly used Kimura two-parameter (K2P) inter-species distance plus four more stringent measures of taxon distinctiveness, (1) Rosenberg's reciprocal monophyly, (P(AB)),¹ (2) Rodrigo's (P(randomly distinct)),² (3) genealogical sorting index, ( gsi),³ and (4) General mixed Yule- coalescent (GMYC).4,5 For both insect datasets, a comparative analysis of the methods revealed that the K2P distance method does not capture the same level of species distinctiveness revealed by the other three measures; in B. tabaci there are more distinct groups than previously identified using the K2P distances and for L. dipsar far less variation is apparent within the predefined subspecies. A consensus for the results from P(AB), P(randomly distinct) and gsi offers greater statistical confidence as to where genetic limits might be drawn. In the species cases here, the results clearly indicate that there is a need for more gene sampling to substantiate either the new cohort of species indicated for B. tabaci or to detect the established subspecies taxonomy of L. dispar. Given the ease of use through the Geneious species delimitation plugins, similar analysis of such multi-gene datasets would be easily accommodated. Overall, the tip to root approach described here is recommended where careful consideration of species delimitation is required to support crucial biosecurity decisions based on accurate species identification. [ABSTRACT FROM AUTHOR]

Summary 1. When an invasive species first colonizes an area, there is an interval before any host-specific natural enemies arrive at the new location. Population densities of newly invading species are low, and the spatial and temporal interactions between spreading invasive species and specific natural enemies that follow are poorly understood. 2. We measured infection rates of two introduced host-specific pathogens, the entomophthoralean fungus Entomophaga maimaiga and the baculovirus Lymantria dispar nucleopolyhedrovirus ( LdNPV), occurring in spreading populations of an invasive forest defoliator, L. dispar (gypsy moth), in central Wisconsin. 3. Over 3 years, we found that host density was closely associated with the presence and prevalence of both pathogens. The fungal and viral pathogens differed in the sensitivity of their response as E. maimaiga was present in lower-density host population than LdNPV. 4. We examined the relationship between weather conditions and infection prevalence and found that activity of both the fungus and virus was strongly seasonally influenced by temperature and rainfall or temperature alone, respectively. 5. Purposeful releases of pathogens (median distances of study sites from release sites were 65·2 km for E. maimaiga and 25·6 km for LdNPV) were not associated with pathogen prevalence. 6. A generalist fly parasitoid, Compsilura concinnata, also killed L. dispar larvae collected from the study sites, and parasitism was greater when infection by pathogens was lower. 7. Our results demonstrated that although infection levels were low in newly established host populations, host-specific pathogens had already moved into host populations close behind advancing populations of an invasive host; thus, spreading hosts were released from these enemies for only a relatively short time. [ABSTRACT FROM AUTHOR]