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Of four diets tested, one based on lima beans and casein was selected as the best and easiest for rearing third instars of Japanse beetles, Popillia japonica Newman. The diet gave the best survival and heaviest pupae and was previously used for rearing larvae of the New Zealand grass grub, Costelytra zealandica (White).

Although Maryland has a long history of the use of biological control, nowhere are these interesting and sometimes landmark efforts summarized. From 1881 and 1992, 144 species of natural enemies were released in Maryland against at least 49 arthropod pests and weeds. Some type of biological control project has been undertaken in each of Maryland′s 23 counties. This paper provides details primarily of the importation, augmentation, and recolonization, into Maryland, of natural enemies of insect and mite pests of plants, as well as of natural enemies of pest plants (i.e., of weeds).

Mortality of last-instar Japanese beetles, Popillia japonica Newman, was highest when exposed to isazophos, bendiocarb, and chlorpyrifos in 30-ml arenas containing California soils in laboratory assays. Carbaryl and two nematodes, Heterorhabditis bacteriophora Poinar and Steinernema glaseri (Steiner), were less effective treatments in these arenas. Soil × treatment (insecticides and nematodes) interactions were highly significant, often changing larvicide rank order of efficacy within soils. Soil characteristics identified by regression analysis as affecting insecticidal activity were organic matter (chlorpyrifos, bendiocarb, and S. glaseri ) and pH (carbaryl and isazophos). In arenas (7.6-cm diameter by 15 cm) simulating turf conditions, S. glaseri was the most effective treatment, followed by chlorpyrifos, then bendiocarb. The influence of irrigation on improving insecticide efficacy increased with higher soil organic matter. Because no treatment was completely effective against larvae, eradication of Japanese beetles cannot depend entirely on control tactics directed against the immature stages.

Feeding preferences of Japanese beetle, Popillia japonica Newman, and southern masked chafer, Cyclocephala lurida Bland, grubs for six common cool-season turfgrasses were evaluated in choice tests in the greenhouse. On the basis of larval distributions, Popillia japonica consistently preferred perennial ryegrass, Lolium perenne L., over all other turfgrasses tested. In contrast, C. lurida showed no consistent pattern of preference. Presence of one grub species did not affect distribution of the other species. Grubs did not discriminate between tall fescue, Festuca arundinacea Schreb., infected with the endophyte Acremonium coenophialum Morgan-Jones & Gams and endophyte-free tall fescue.

Several trap designs and trap heights were tested on a golf course for effectiveness of capture of Japanese beetle ( Popillia japonica Newman) adults. A single Trece trap top or four Trece trap tops (each with quarter-strength lures) on 121-liter trash container collection bases caught significantly more beetles than a standard Trece trap or a large vane trap on a 121-liter container. Traps with a single Trece trap top on a 121-liter container also caught significantly more beetles than a standard Trece trap when both types were emptied daily to determine if captured beetles were influencing trap capture. Significantly more beetles were captured by traps with trap collection funnel rims at 13 cm compared with rims at 90 cm above the ground.

Infective juveniles of the entomopathogenic nematode Heterorhabditis bacteriophora Poinar (Cl strain) were applied to Kentucky bluegrass (Poa pratensis L.) to determine the effect of irrigation on efficacy against larvae of the Japanese beetle, Popillia japonica Newman, and the northern masked chafer, Cyclocephala borealis Arrow. Excellent control (>;80%) was achieved with 1.25-5.0 billion nematodes per hectare when plots received irrigation before treatment application and again within 24 h after treatment application. When plots were not irrigated before treatment, results were inconsistent (range, 0-92% of the control). Two spray volumes (0.16 liters/m2 and 0.32 liters/m2) had no significant effect on nematode efficacy.

The effects of agroecosystem diversification on the most abundant herbivore species of soybean ( Glycine max Merrill) in Ohio were monitored during 1988, 1989 and 1990. Maize and soybean plots were arranged in a randomized complete block design with four replicates for a 3 × 2 factorial experiment. Factors were cropping systems (maize monoculture, soybean monoculture and strip intercropping of maize and soybean) and tillage systems (no-tillage and conventional tillage). The herbivore fauna of soybean was sampled by sweep net and suction net (D-Vac). Statistically significant results from at least 1 year indicated that no-tillage was favorable to Japanese beetle ( Popillia japonica Newman), grape colaspis ( Colaspis brunnea (F.)), tarnished plant bug ( Lygus lineolaris (Palisot de Beauvois)), homopterans (Cicadellidae and Cercopidae), grasshoppers ( Melanoplus spp.) and slugs (Gastropoda). Potato leafhopper ( Empoasca fabae (Harris), green cloverworm ( Plathypena scabra (F.)), and striped flea beetles ( Phyllotreta spp.) were more abundant in conventional tillage plots. Strip intercropping was favorable to green cloverworm, Japanese beetle, and grape colaspis, whereas the potato leafhopper were more abundant in monoculture plots. There was no evidence of reduced herbivory in diversified agroecosystems, as predicted by the resource concentration hypothesis.

Two methods of subsurface placement technology, high-pressure liquid injection (HPLI) and an adapted overseeder (OS), were used to apply isazophos (Triumph) to golf course roughs that were infested with Japanese beetle ( Popillia japonica Newman) grubs. In every instance (five trials with an emulsifiable concentrate formulation, one trial with a granular formulation), isazophos applied subsurface at 1.1 or 0.6 kg (AI)/ha performed as well or better than isazophos applied conventionally on the surface at 2.2 k (AI)/ha. This apparent enhancement of efficacy of isazophos should provide turf managers with an alternative that can reduce the amount of insecticide on turf, at least under certain field conditions. In addition, irrigation after application on plots treated with HPLI appeared to be less critical to efficacy of isazophos than on plots treated conventionally.

A strain of Steinernema carpocapsae selectively bred in the laboratory for improved host-finding of scarab larvae did not provide enhanced field efficacy of Japanese beetle larvae, Popillia japonica. Selected (S20) and wildtype (All) strains reduced larval populations by 25-30%, compared with 70% for the chemical insecticide. Similar results were obtained in laboratory exposures conducted in soil-filled pots against the scarab Maladera matrida . The selected S20 strain was, nevertheless, superior to the All and HP88 strains at locating larvae. S20 nematodes, which are known to have enhanced chemosensitivity to carbon dioxide, were observed to aggregate heavily at or near the spiracles. Scarab spiracles are covered with sieve plates and, therefore, are not a portal of nematode entry. Despite selection for greater response to host cues, S20 persisted in nictating, standing on their tails, even when a host was present. The unfavorable field results usually obtained with S. carpocapsae against scarab larvae have been attributed to low powers of host search. Our results indicate that even when larvae are located S. carpocapsae is poorly adapted to cause infection. Genetic improvement cannot he expected to overcome poor adaptation. Future selection efforts with nematodes should begin with species adapted for the target host.

The Japanese beetle (JB), Popillia japonica , exhibits rapid paralysis after consuming flower petals of zonal geranium, Pelargonium x hortorum . Activity-guided fractionations were conducted with polar flower petal extracts from P. x hortorum cv. Nittany Lion Red, which led to the isolation of a paralysis-inducing compound. High-resolution–MS and NMR ( 1 H, 13 C, COSY, heteronuclear sequential quantum correlation, heteronuclear multiple bond correlation) analysis identified the paralytic compound as quisqualic acid (C 5 H 7 N 3 O 5 ), a known but rare agonist of excitatory amino acid receptors. Optical rotation measurements and chiral HPLC analysis determined an l -configuration. Geranium-derived and synthetic l -quisqualic acid demonstrated the same positive paralytic dose–response. Isolation of a neurotoxic, excitatory amino acid from zonal geranium establishes the phytochemical basis for induced paralysis of the JB, which had remained uncharacterized since the phenomenon was first described in 1920.

Field studies were conducted in Virginia cornfields from 1989 to 1991 to evaluate a preplant method for detecting damage to germinating corn seeds by multiple species of insect pests, including seedcorn maggots, Delia platura (Meigen); several species of wireworms in the genera Melanotus, Conoderus , and Aeolus ; and several species of white grubs in the genera Popillia, Cyclocephala, Cotinis , and Maladera . The method used consisted of evaluating feeding damage to com seeds in wire-mesh traps (i.e., baited wire traps) that were buried in cornfields for ≍2 wk before planting. In the 1989 study conducted in a cornfield with a large infestation of wireworms, a significantly greater proportion of com seeds exhibited feeding damage in the baited wire traps (38.3%) compared with corn seeds planted by hand at conventional plant spacings (i.e., simulated commercial plantings) (31.5%). Further evaluation in 47 cornfields in 1990 and 1991 revealed no significant difference between the proportion of damaged corn seeds in the baited wire traps (2.7%) and simulated commercial plantings (2.0%). A Significantly greater proportion of com seeds exhibited feeding damage in the commercial plantings (5.4%) compared with the baited wire traps; however, mean stand loss associated with insect feeding in the commercial plantings was only 0.4%, which was substantially less than the proportion of damaged com seeds in the haited wire traps. Significant linear regressions were obtained between the proportion of damaged seeds in the baited wire traps and the proportion of damaged seeds in the simulated commercial plantings, commercial plantings, and proportion of stand loss in the commercial plantings. However, the lack of economic stand loss in the commercial plantings and low regression coefficient of determination precluded the development of a baited wire trap damage threshold to predict stand loss in fields subsequently planted with corn.

The entomopathogenic nematodes Steinernema carpocapsae (Weiser) (All, S17, and S20 strains), S. feltiae (Filipjev) (= S. bibionis) (N27 strain), Heterorhabditis bacteriophora (Poinar) (HP88 strain), and Heterorhabditis sp. (Bacardis strain) were evaluated as biological control agents of the sweetpotato weevil, Cylas formicarius (F.), during two consecutive growing seasons in southern Florida. Two strains, S17 and S20, were genetically selected over 17 and 20 generations, respectively, for enhanced finding of the host scarab Popillia japonica Newman. Steinernematid nematodes were applied as aqueous suspensions. The heterorhabditid nematodes, tested only in the second year, were applied using cadavers of greater wax moth, Galleria mellonella (L.), infected with these nematodes. Abundance and damage of weevils differed among nematode treatments. In the first experiment, S. carpocapsae All and S17 were most efficacious at reducing weevil damage and produced the highest percentages of marketable roots. In the second experiment, the heterorhabditid nematode, HP88, was most efficacious at reducing weevil damage to storage roots. All of the steinernematids, except S20 applied at a high rate (7.9 billion infective juveniles per ha), produced comparable percentages of marketable roots but were less efficacious than HP88. In the first experiment, S. carpocapsae All and S17 persisted poorly, whereas S. feltiae N27 persisted for >160 d after application. In the second experiment, most nematodes were recovered at moderate levels for >120 d after application; however, only Heterorhabditis sp. Bacardis was consistently recovered at high levels for >230 d after application.

Penetration of infective juveniles of Steinernema glaseri and Steinernema carpocapsae into Japanese beetle larvae, Popillia japonica , was compared using external exposure and injection methods. Steinernematids entered the grubs through the mouth and anus, but not through the spiracles or cuticle. For all treatments, S. glaseri had higher levels of penetration than S. carpocapsae . Penetration of the gut by orally injected nematodes began 4-6 hr postinoculation. All regions of the gut were penetrated, with more than 82% of all penetrating nematodes invading the host hemocoel via the midgut. Gastric cecae were important sites for nematode penetration. Penetration sites of anally injected nematodes were the rectal sac of the hindgut. Penetration rates of isolated guts and that in vivo did not differ. Defecation by P. japonica was rapid, with orally injected nematodes appearing in the feces in 4-8 hr. Our results indicate that compared with S. glaseri, S. carpocapsae is poorly adapted to parasitize Japanese beetle larvae.

The pheromones (1a,1b) of two insect pests, the Japanese Beetle (Popillia japonica, Newman) and Cupreus Chafer Beetle (Anomala cuprea, Hope), were synthesized. Starting from protected D-Ribose, 1a was prepared in 8 steps with a 30% total yield. The same chemistry was demonstrated for the synthesis of 1b.

Distribution of Japanese beetle, Popillia japonica Newman, grubs and dispersal of an entomopathogenic nematode, Heterorhabditis bacteriophora Poinar ‘Oswego’ strain (an isolate from New York state), were examined for 5 wk within soil-filled flats containing grass. Japanese beetle grubs uniformly dispersed to all sections of the flats not infested with H. bacteriophora ‘Oswego’ strain. In flats infested with H. bacteriophora ‘Oswego’ strain, however, greater proportions of Japanese beetle grubs were recovered in sections near the nematode release site or center sections of the flats. H. bacteriophora ‘Oswego’ strain dispersed to all sections of the flats but dispersed more rapidly within the flats infested with Japanese beetle grubs than in flats not infested with Japanese beetle grubs.

Control of Japanese beetle ( Popillia Japonica Newman) larvae by Heterorhabditis Bacteriophora Poinar (HP88 strain and a New Jersey strain, NJ-2) and Steinemema glaseri Steiner (NC strain and a New Jersey strain, NJ-43) showed that all strains reduced larval populations to a level comparable with that achieved by use of bendiocarb. Nematode strains and species did not differ significantly in mean level of population reductions ( H. bacteriophora HP88 and NJ-2, 51.0 and 71.6%; S. glaseri NC and NJ-43, 50.4 and 70.1%, respectively). However, variation in the level of population reduction provided by NJ-43 was less than that by the NC strain. After a 3-wk test period, nematode persistence and downward migration in the field did not differ among species and strains. In a laboratory bioassay, New Jersey strains provided higher levels of larval mortality compared with strains cultured in the laboratory for at least a decade. A new method used to count viable bacteria retained by individual infective juveniles showed that NJ-43 not only retained more bacteria than the NC strain (19.5 ± 0.2 compared with 4.9 ± 0.06 bacteria per infective juvenile) but also that the proportion of nematodes retaining bacteria was higher (77% compared with 55%). Bacterial retention by infective juveniles did not differ significantly between H. bacteriophora strains.

The pheromones (1a,1b) of two insect pests, the Japanese Beetle (Popillia japonica, Newman) and Cupreus Chafer Beetle (Anomala cuprea, Hope), were synthesized. Starting from protected D-Ribose, 1a was prepared in 8 steps with a 30% total yield. The same chemistry was demonstrated for the synthesis of 1b.

Because mating may be costly, sexually active males or females are predicted to be in relatively good physiological condition and may preferentially direct their mating behavior toward relatively high-quality mates. We tested this hypothesis in Japanese beetles (Popillia japonica New- man), a pest species in which males and females may be either isolated or in aggregations while feeding on host plants. We examined male size and lipid content and female size and egg load with respect to both their pairing status and whether they were isolated or in aggregations. Males that were paired had the highest lipid levels, and single, isolated males had the lowest. Paired females had the highest egg loads and single, isolated females had the lowest. Neither male nor female size was related to pairing status. Females captured during the times of relatively high pairing frequency (i.e., morning and evening) had higher egg loads than females captured at times of lower pairing frequency (i.e., afternoon). These results suggest that mating and aggregative behaviors in Japanese beetles are dependent on the physiological status of males and the reproductive condition of females.

Japanese beetles, Popillia japonica Newman (Coleoptera: Scarabaeidae), were evaluated for ovipositional preferences among four turfgrasses common in northwestern Arkansas. Choice assays revealed females preferred to oviposit in tall fescue (Festuca arundinacea Schreb.) and zoysiagrass (Zoysia japonica Steud.), and that they avoided oviposition in common bermudagrass (Cynodon dactylon [L.] Pers.) and hybrid bermudagrass (C. dactylon x C. transvaalensis Pers.). Significantly fewer eggs were oviposited in hybrid bermudagrass in a no-choice assay, suggesting that chemical and/or physical plant characteristics deter oviposition in that grass. The percentage of turfgrass cores with evidence of female activity (presence of female or eggs, or signs of female digging) in choice assays revealed no differences among treatments, yet significantly fewer hybrid bermudagrass cores had eggs. These results suggest that many females did not initially reject hybrid bermudagrass based on aboveground plant characteristics, but rather they left without ovipositing. Therefore, resistance in hybrid bermudagrass is likely expressed below ground. Our results suggest that the use of hybrid bermudagrass as a means of cultural control in an integrated pest management program may discourage Japanese beetle oviposition and subsequent grub infestations in lawns, golf courses, or sports fields.

In choice tests, Japanese beetles, Popillia japonica Newman, laid more eggs the wetter the soil, up to about field capacity. No eggs were laid in soils with less than or equal to 55% moisture content. Even in nonchoice tests, Japanese beetles laid few eggs in dry soils. In choice tests, Japanese beetles laid more eggs in soils with medium clay contents than other soil textures. In nonchoice tests, Japanese beetles laid similar numbers of eggs into a wide range of soil textures, except fewer eggs were laid into pure sand. In choice tests, rose chafer, Macrodactylus subspinosus (F.), also laid many more eggs in wetter soil than in drier soil. In choice tests, rose chafers showed no preference for oviposition in any of the soil textures tested. These data help explain the patchy distribution of Japanese beetle larvae and of rose chafer adults after dry summers, but do not explain reported occurrences of rose chafer larvae only in sandy soil.

Sampling statistics were determined for Japanese beetles, Popillia japonica Newman, caught in traps baited with a 3:7:3 mixture of phenethyl propionate/eugenol/geraniol, or with ( R, Z )-5-(1-decenyl)dihydro-2(3 H )-furanone (Japonilure), or with a combination of both lures. Iwao’s patchiness regression and a quadratic model were inappropriate for modeling the mean-variance relationships. Taylor’s power law was used to model these data, and relationships were developed for beetle numbers captured by traps baited with phenethyl propionate-eugenol-geraniol alone or in combination with Japonilure and for males and females captured by traps baited with Japonilure alone. Relationships to determine sample sizes for fixed levels of precision were also developed. The I n ( X + 1) transformation did not standardize the mean-variance relationship for any data set, but the Healy & Taylor transformation was effective on all data sets.

Several strains or species of entomopathogenic nematodes-Steinernema carpocapsae Weiser (All), S. feltiae (= bibionis ) (Filipjev) (Biosys strains 27 and 980), S. glaseri (Steiner), and Heterorhabditis bacteriophora (= heliothidis ) Poinar (HP88)—were tested in the field for control of Japanese beetle, Popillia japonica Newman; Oriental beetle, Anomala orientalis Waterhouse; or black turfgrass ataenius, Ataenius spretulus (Haldeman) larvae. A spring field trial against A. orientalis , in which half of the plots were aerated to assist in nematode dispersal in soil, showed no significant differences between control plots and those treated with H. bacteriophora (HP88) or S. carpocapsae (All). A fall field trial in which half of the plots were dethatched before nematode application (to enhance nematode-larva contact) also showed no significant differences between plots treated with S. feltiae and control plots. Four other fall field trials with S. carpocapsae, S. feltiae, S. glaseri , and H. bacteriorphora against Japanese beetle larvae showed a range of 0-81% control. The greatest mortality of P. japonica larvae occurred with S. glaseri at 24.7 × 109 per ha. A field trial with S. carpocapsae against A. spretulus showed 94% mortality at a 24.7 × 109 per ha rate.

Preference of adult Japanese beetle (Popillia japonica Newman) for different species, varieties, and cultivars was compared among 33 crabapple (Malus Mill.), nine cherry (Prunus L.), and nine birch (Betula L.) taxa. Field-grown crabapples and cherries and container-grown birches were visually rated based on the percentage of total leaf area skeletonized by natural populations of this beetle. The largest variation in the preference of Japanese beetles was found among the crabapples, with injury ranging from 0 to 83%. Eighteen of the crabapple taxa had no significant injury. All of the cherry taxa were skeletonized in excess of 46%, suggesting little natural resistance among these plants. Eight of the birch taxa had no significant injury, while one species, B. jacquemontii, had an average injury of 16%. An additional study was conducted to evaluate the degree of resistance of selected crabapple taxa by caging beetles on branches of individual trees in no choice tests. Although beetles continued to feed on one resistant species, M. hupehensis (a species without injury under natural conditions), beetles would not feed on leaves of another resistant cultivar, M. baccata ‘Jackii’, when caged on those trees. These results indicate there is considerable potential for using resistance to adult Japanese beetle feeding as one selection criterion when choosing taxa of crabapple and birch for use in the landscape. Results also suggest that the mechanism(s) of resistance can vary and may include aspects of avoidance (low attraction of beetles) as well as antixenosis.

Billions of nematodes were released from 1939 to 1942 throughout the state of New Jersey (563 sites) in an effort to colonize the entomopathogenic species Steinernema glaseri for biological control of the Japanese beetle, Popillia japonica. Because of the onset of World War 11 and the postwar development of chlorinated hydrocarbon insecticides, little effort was expended to evaluate the outcome of these introductions. We evaluated this colonization program by collecting soil samples in 1991 from 304 geographically and ecologically diverse sites across New Jersey. The soil samples were assayed for entomopathogenic nematodes using the Galleria bait method. Overall, 66 (21.7%) soil samples were positive for entomopathogenic nematodes: 24 steinernematids and 42 heterorhabditids. The most common species isolated was Heterorhabditis bacteriophora (38 isolates), followed by S. glaseri (14), S. carpocapsae (4), S. feltiae (4), Heterorhabditis spp. (4), and Steinernema spp. (2). S. glaseri was recovered only from the southernmost third of the state. We conclude that the colonization effort initiated more than 50 years ago was unsuccessful. The reasons remain uncertain, but intolerance of S. glaseri to temperate climates is one likely explanation. That is, southern New Jersey appears to represent the northernmost range of this neotropical species. Moreover, early workers were unaware of the nematode's mutualistically associated bacterium, Xenorhabdus poinarii, which plays important roles in killing insect hosts and in nematode reproductive potential. We show that the bacterium is inhibited by antimicrobial compounds used by these workers during mass rearing, so it is probable that only the nematode portion of the nematode-bacterium complex was released.

Field tests were conducted in 1986 and 1987 to evaluate control of Japanese beetle larvae, Popillia japonica Newman, in turfgrass plots by heterorhabditid and steinemematid nematodes. Heterorhabditis bacteriophora Poinar (NC strain) reduced populations a maximum of 60%, 34 d after a fall 1986 treatment. Control increased to 96% before pupation the following spring, and was 93-99% of the next larval generation. Application of Steinemema (= Neoaplectana ) carpocapsae Weiser (All strain) provided a maximum of 51% control after 34 d, 90% the next spring (290 d after treatment), and 0% after 386 d. A spring 1987 application of H. bacteriophora resulted in 68% control 28 d after treatment, and up to 67% of the following generation of] apanese beetle larvae in the fall (138 dafter treatment). H. bacteriophora (HP88 strain) gave 100% control after 28 d in the spring of 1987, and 93-97% control of the next generation of larvae. No adverse effects on nontarget organisms (mites or collembola) were observed 28 d after treatments with any of the nematodes. Our data show that nematodes reproduce in Japanese beetle larvae, survive in the field in turfgrass, and have an influence on target hosts for a longer period oftime than previously demonstrated.

Suitability of six cool-season turfgrasses, including tall fescue, festuca arundinacea Schreb., infected or not infected by the endophyte Acremonium coenophialum Morgan-Jones & Gams, was studied for root-feeding grubs of the Japanese beetle, Popillia japonica Newman, and the southern masked chafer, Cyclocephala lurida Bland. Larval growth, survival, food use, effects of larval diet on adult weight and fecundity, and natural incidence of grubs were investigated in laboratory, greenhouse, and field assays. Hard fescue, Festuca ovina var. duriuscula L., endophyte-free tall fescue, and perennial ryegrass, Lolium perenne L., were generally more favorable than creeping bentgrass, Agrostis palustris (Huds.), for growth of C. lurida, whereas Kentucky bluegrass, Poa pratensis L., was a relatively poor host for both grub species. Variation in nitrogen content of roots is apparently too small to explain these differences. Roots of endophyte-infected tall fescue contained 93 micrograms/g N-formyl loline, a level previously shown to deter grub feeding on artificial medium. However, this study suggests that antixenosis may not occur in the presence of the stimulus complex in tall fescue roots. Survival or growth or both of neonate first instars were lower on endophyte-infected than on endophyte-free tall fescue. However, response of older instars was variable, and there was no measurable effect of endophyte on density or weight of grubs in field plots or on fecundity of P. japonica adults that emerged from endophyte-infected turf. Variation in susceptibility of cool-season turfgrasses to white grubs is probably affected more by differences in their ability to tolerate the feeding damage than by their inherent suitability as food.

A newly isolated strain, designated Buibui, belonging to Bacillus thuringiensis serovar japonensis had potent larvicidal activity against the scarabaeids Anomala cuprea Hope and Popillia japonica Newman. Preliminary tests indicated that other scarabaeid larvae including A. albopilosa Hope, A. rufocuprea Motschulsky, A. daimiana Harold, A. schonfeldti Ohaus, Mimela splendens (Gyllenhal), and Blitopertha orientalis (Waterhouse) were equally susceptible to strain Buibui. However, the strain showed no insecticidal activity against larval lepidopteran insects including Bombyx mori (L.) (Bombycidae), Plutella xylostella (L.) (Plutellidae), Spodoptera litura Fabricius, S. exigua Hubner (Noctuidae), and Adoxophyes sp. (Tortricidae) and coleopterans, Allomyrina dichotoma L. (Scarabaeidae) and Henosepilachna vigintioctopunctata Fabricius (Coccinellidae). At 14 days post-treatment, the LC50 and LC95 of the δ-endotoxin against the cupreous chafer, A. cuprea, were estimated by probit analysis as 0.098 ± 0.019 and 0.29 ± 0.086 μg protein/g compost, respectively. Strain Buibui showed no β-exotoxin activity.

RH 5849 (1,2-dibenzoyl-1-tert-butylhydrazine), a nonsteroidal ecdysone agonist representative of a novel class of insect growth regulators, was tested against larval and adult Japanese beetles, Popillia japonica Newman; and fall armyworm, Spodoptera frugiperda J.E. Smith, caterpillars in turf. RH 5849 was effective against Japanese beetle grubs at concentrations as low as 1 ppm in soil. Symptoms of toxicity included discoloration, weight loss, apparent neurotoxic effects, cessation of feeding, and developmentally premature, lethal molting at higher rates. Sublethal effects on overwintered third instars included earlier formation of prepupae. Against second-instar grubs, RH 5849 was nearly as effective as isazophos (Triumph 4E) at comparable rates in the field. Subsequent egg production by female beetles that fed once on RH 5849-treated sassafras foliage was reduced by 66-74%, but hatching success of eggs was not affected. RH 5849 caused premature molting and 100% mortality of fall amlyworms fed tall fescue foliage that wassprayed once with 30 or 100 ppm, or treated systemically through the roots. Systemic activity of RH 5849 is noteworthy because it could provide simultaneous control of both root-feeding and foliar-feeding turfgrass pests.

Ovavesicula popilliae is a microsporidian pathogen used as a biological control agent of the Japanese beetle ( Popillia japonica Newman), and reportedly produces 32 spores within a sporophorous vesicle measuring 20.0-21.0 μm × 15.0-15.5 μm. We determined the sporophorous vesicles to have a mean size of 13.3 μm × 9.3 μm and each with a mean of 24.7 spores. Varying number of spores may be due to failed divisions of sporonts, death of spores after formation, host effects or different pathogen strains. Ovavesicula popilliae es una microsporidio patogeno utilizado como agente de control biologico del escarabajo japones ( Popillia japonica Newman), que segun reportes produce 32 esporas dentro de vesiculas con medidas de 20.0-21.0 μm × 15.0-15.5 μm. Determinamos que las vesiculas tienen un tamano de 13.3 μm × 9.3 μm y una media de 24.7 esporas. La variacion en el numero de esporas puede deberse a fallas en la division de los sporonts, muerte de las esporas despues de su formacion, efectos del hospedero, o diferencias en las cepas del patogeno. View this article in BioOne

A spore-forming bacterium isolated from the soil of Japan was assigned to Bacillus thuringiensis serovar japonensis (flagellar antigen 23). Parasporal inclusions of this isolate were spherical to ovoid in shape and exhibited high larvicidal activity against coleopterous scarabaeid beetles, the cupreous chafer. Anomala cuprea, the soybean beetle, Anomala rufocuprea, and the Japanese beetle. Popillia japonica. No toxicity was shown by this isolate against larvae of Lepidoptera, Diptera, and Orthoptera, and adults of a chrysomelid coleopteran.

Soybean [Glycine max (L.) Merr.] was one of the most important legume crops in the world in 2010. Japanese beetles (JB; Popillia japonica, Newman) in the US were an introduced and potentially damaging insect pest for soybean. JBs are likely to spread across the US if global warming occurs. Resistance to JB in soybean was previously reported only in plant introductions. The aims here were to identify loci underlying resistance to JB herbivory in recombinant inbred lines (RILs) derived from the cross of Essex × Forrest cultivars (EF94) and to correlate those with loci with factors that confer insect resistance in soybean cultivars. The RIL population was used to map 413 markers, 238 satellite markers and 177 other DNA markers. Field data were from two environments over 2 years. Pest severity (PS) measured defoliation on a 0–9 scale. Pest incidence (PI) was the percentage of plants within each RIL with beetles on them. Antibiosis and antixenosis data were from feeding assays with detached leaves in petri plates. Five QTL were detected for the mean PS field trait (16%

Forty-one plant essential oils were tested under field conditions for the ability to reduce the attraction of adult Japanese beetles, Popillia japonica Newman (Coleoptera: Scarabaeidae), to attractant-baited or nonbaited traps. Treatments applied to a yellow and green Japanese beetle trap included a nonbaited trap, essential oil alone, a Japanese beetle commercial attractant (phenethyl proprionate:eugenol:geraniol, 3:7:3 by volume) (PEG), and an essential oil plus PEG attractant. Eight of the 41 oils reduced attractiveness of the PEG attractant to the Japanese beetle. When tested singly, wintergreen and peppermint oils were the two most effective essential oils at reducing attractiveness of the PEG attractant by 4.2x and 3.5x, respectively. Anise, bergamont mint, cedarleaf, dalmation sage, tarragon, and wormwood oils also reduced attraction of the Japanese beetle to the PEG attractant. The combination of wintergreen oil with ginger, peppermint, or ginger and citronella oils reduced attractiveness of the PEG attractant by 4.7x to 3.1x. Seventeen of the 41 essential oils also reduced attraction to the nonbaited yellow and green traps, resulting in 2.0x to 11.0x reductions in trap counts relative to nonbaited traps. Camphor, coffee, geranium, grapefruit, elemi, and citronella oils increased attractiveness of nonbaited traps by 2.1x to 7.9x when tested singly, but none were more attractive than the PEG attractant. Results from this study identified several plant essential oils that act as semiochemical disruptants against the Japanese beetle.

Traps containing a combination floral and synthetic pheromone lure are used to monitor and manage Japanese beetles, Popillia japonica Newman (Coleoptera: Scarabaeidae). One key factor limiting trap effectiveness for beetle control is the “trap spillover” phenomenon, in which the trap attracts beetles without capturing them, resulting in increased damage to surrounding host plants. We investigated the mechanisms underlying trap spillover by conducting two studies in a soybean field in east central Illinois. In the first study, we set up trap stations for 1 d and compared the sex, size, and egg load (for females) of beetles caught in the traps with those on the plants immediately surrounding the trap, downwind of the trap, at lure-only (no trap) stations, and at control areas. Females caught in traps tended to be smaller than those on plants surrounding the traps, and females attracted to the traps had fewer eggs than those downwind or at control sites. We did not find any difference in male characteristics. In the second study, we observed the behavior of beetles initially approaching traps. Upon initial approach, the majority of individuals landed on plants before making contact with the trap, and those beetles that spent an extended time on the leaves tended to be females. Arriving males would occasionally pair with these females on the plants. Overall, traps did not capture a random subset of the beetles present in the field. We hypothesize that trap spillover is a result of arriving females not being as attracted to the precise location of the trap as they are to the general location itself, and of arriving males seeking mates and finding them among these spillover females.

BACKGROUND: Japanese beetles, Popillia japonica Newman, are a quarantine challenge for nursery shipments from infested to non-infested states. Marathon (imidacloprid) and Discus (imidacloprid + cyfluthrin) are approved preharvest nursery treatments (US Domestic Japanese Beetle Harmonization Plan; DJHP). This study evaluated approved and non-approved (acephate, carbaryl, clothianidin, dinotefuran, halofenozide, thiamethoxam, trichlorfon) preharvest treatments, optimal rates (labeled 1×, 0.3–0.75× or 2–3×) and optimal timings (June, July, August and September) to control early-instar (grubs) P. japonica in field nurseries. RESULTS: Most insecticides effectively reduced grub densities, except for acephate, carbaryl and trichlorfon. Clothianidin, thiamethoxam and halofenozide provided grub control equivalent to DJHP standards during most years. Across all test years and timings, percentage grub reductions were: Marathon (1×: 59.2–100; 3×: 78.9–100), Discus (1×: 60.7–100), clothianidin (1×: 96.1–100; 3×: 97.4–100), thiamethoxam (1×: 75.0–100; 3×: 80.0–100), halofenozide (1×: 70.0–100; 3×: 90.0–100) and dinotefuran (1×: 13.2–88.2; 3×: 71.1–93.4). CONCLUSIONS: July application was more consistent and effective than other timings, and higher rates (2× and 3×) did not generally improve grub control. Overall, clothianidin, thiamethoxam and halofenozide (and dinotefuran applied in August) were equivalent to current DJHP standards. These insecticides may be suitable for DJHP Category 2 states, potentially lowering grower costs. Copyright © 2009 Society of Chemical Industry

Publisher Summary This chapter focuses on the Japanese beetle, Popillia japonica Newman. It is among the most polyphagous of plant-feeding insects. The adults skeletonize the foliage or feed on the flowers or fruits, of nearly 300 species of wild or cultivated plants. The larvae, or grubs, develop in the soil where they feed on roots of turf and pasture grasses, vegetables, nursery seedlings, and field crops. It belongs to the family Scarabaeidae, subfamily Rutelinae. Adults are broadly oval, 8–11 mm in length, metallic green, with coppery-brown elytra that do not quite cover the end of the abdomen. The abdomen bears five patches of white hairs on both side, and another pair near its tip. Females, which tend to be slightly larger than males, have an elongate, spatula-shaped spur on the fore tibia, used for digging. This spur is shorter and pointed in males. Japanese beetles have a 1-year life cycle in most parts of their range. Adults occur from June to August. Upon emergence from the soil, virgin females emit a volatile sex pheromone that attracts clusters of males. Subsequent mating occurs on food plants. The beetles typically feed from the upper surface of leaves, chewing out the tissue between the veins and leaving a lace-like skeleton.

The effect of culture method on the efficacy of entomopathogenic nematodes was assessed by analysis of 511 greenhouse and field trials. In tests against larvae of the Japanese beetle, Popillia japonica Newman, and black vine weevil, Otiorhynchus sulcatus (F.), infective juveniles of Steinernema carpocapsae (Weiser) reared by in vivo, in vitro solid, and in vitro liquid culture methods provided equivalent larval reductions. Heterorhabditis bacteriophora Poinar infective juveniles produced in vivo and on solid media provided equivalent results in trials against Japanese beetle larvae and northern masked chafer, Cyclocephala borealis (Arrow). By contrast, H. bacteriophora produced by liquid culture achieved significantly lower Japanese beetle reductions than those reared in vivo or on solid media. Against the northern masked chafer, liquid culture H. bacteriophora provided significantly less host mortality than solid culture nematodes. Field persistence studies provided further evidence that liquid culture H. bacteriophora were inferior to infective juveniles produced on solid media. The poor performance of liquid culture H. bacteriophora was attributed to low lipid assimilation.

The effect of soil temperature and water on fungus-induced mortality of scarab grubs was investigated in the laboratory. Soil applications of dry mycelial particles of Metarhizium anisopliae (Metsch.) Sorokin were tested against the Japanese beetle, Popillia japonica Newman, and particles of Beauveria brongniartii (Saccardo) Petch were tested against the European chafer, Rhizotrogus majalis (Razoumowsky). Japanese beetle mortality occurred fastest in soil at 27°C and 11% water content and slowest in soil at 21°C and 17.5% water content. Japanese beetle mortality at Week 5 of the bioassay was affected by both soil temperature and water when a high dosage of fungus was used. The proportion of Japanese beetle cadavers with sporulating fungus was influenced by soil water, dose, and a soil temperature and dose interaction. In contrast, European chafer mortality was affected only by soil temperature. Mortality rates for B. brongniartii -killed European chafers and the proportion of cadavers supporting sporulating fungus were both significantly higher in soil at 21°C than at 27°C. Survival of M. anisopliae and B. brongniartii was affected by soil water; concentrations of both fungi were consistently higher in soil at low water content.

We tested two agronomic practices that are likely to increase plant and structural diversity, no-tillage and strip intercropping, for effects on corn invertebrate fauna. Some of the most common herbivores and natural enemies were sampled by direct counts and damage estimation from 1988 through 1990 on monoculture corn and strips of corn alternated with soybean, under no-tillage and conventional tillage. Among soil pests, cutworms (mostly the black cutworm, Agrotis ipsilon (Hufnagel)); armyworm, Pseudaletia unipuncta (Haworth); and slugs (Gastropoda) were more abundant in no-tillage plots, although only slugs caused severe damage. The western corn rootworm, Diabrotica virgifera virgifera LeConte, and the European corn borer, Ostrinia nubilalis (Hubner), were generally more abundant in conventional tillage plots. Despite crop rotation, the strip-intercropping system (four rows of each crop) was less effective in reducing western corn rootworm infestation, especially in conventional tillage plots. In 1990 only, ladybugs (mostly Coleomegilla maculata (DeGeer)) were more abundant in conventional tillage plots, whereas tarnished plant bugs, Lygus lineolaris (Palisot de Beauvois), were more abundant in notillage plots. Japanese beetle, Popillia japonica Newman; stink bugs, Acrosternum hilare (Say) and Euschistus servus (Say); and spiders (Aranea) were not significantly affected by treatments.

Collections of scarab beetles using chemical lures in Korea consisted of 21 species. The species collected in the greatest numbers were Anomala corpulenta Motschulsky, Anomala sp., Holotrichia morosa Waterhouse, Popillia uchidai Niijimi et Kinoshita and P. indigonacea Motschulsky. Population density and activity varied according to location, with earlier emergence on the southern island of Cheju-do. P. japonica Newman were not captured in Korea even though dense populations are common in Japan and previous publications indicated its presence. However, males of a closely related species, P. uchidai, were present in high numbers throughout the country. These responded to Japanese beetle sex lures and combinations of floral lures, phenethyl propionate and eugenol and geraniol (PEG), much as P. japonica is known to do, but P. uchidai did not respond to PEG, and unlike P. japonica no female response was observed. Anomala corpulenta and Anomala sp. males were attracted to P. japonica sex lures in great numbers. This is the first report of this type of response by beetles outside the genus Popillia to this lure.

The effect of entomopathogenic nematodes on nontarget arthropods in the laboratory, field soils, and a stream were assessed. In the laboratory, adult predators were less susceptible to the nematodes Steinernema carpocapsae (Weiser) (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) than the immature stages. In field tests, entomopathogenic nematodes that had significantly suppressed pest populations ( Popillia japonica Newman, Japanese beetle, Scapteriscus vicinus Scudder, tawny mole cricket, Otiorhynchus sulcatus (F.), black vine weevil, Delia radicum (L.), cabbage maggot, and Diabrotica virgifera virgifera LeConte, western corn rootworm) did not adversely affect the numbers of nontarget soil arthropods in comparison with the untreated control. In contrast, broad-spectrum chemical insecticides (isofenphos, ethoprop, or chlorpyrifos used as chemical checks) significantly reduced or showed a tendency to reduce nontarget arthropod populations. In a stream trial, S. carpocapsae significantly reduced black fly larval populations, but the nontarget insects often increased in the treatment sites. Decreases in nontarget populations were matched by approximately equal or greater reductions in the upstream controls. We conclude that entomopathogenic nematodes do not adversely affect nontarget arthropods when used for short-term control of insect pests.

In an examination of entomopathogenic nematodes used in inundative releases on turfgrass against Japanese beetle larvae, Popillia japonica Newman, 380 treatments from 82 field trials performed from 1984 to 1988 were analyzed using a standard protocol. The results show that most test failures can be explained on the basis of unsuitable nematode strains or environmental conditions. Steinemema carpocapsae (Weiser) appears ill-adapted to parasitize Japanese beetle larvae under any range of conditions. By contrast, the HP88 strain of Heterorhabditis bacteriophora Poinar, produced on solid media, provides control comparable with that by chemical insecticides at the appropriate season (fall), soil temperature (>20°C), soil type (silty clay), irrigation frequency (1-4-d intervals), and thatch depth

The G13 strain of Steinernema carpocapsae (Weiser), previously selected for enhanced host-finding of waxmoth larvae, Galleria mellonella L., was subjected to selection for the same trait against scarab larvae. Twenty rounds of selection produced a 3.7-fold increase, from 18.4 to 68.7%, in nematode (S20 strain) location of nondiapausing larvae of the Japanese beetle, Popillia japonica Newman. However, the overall improvement over wild-types (Foundation and All strains), after 33 rounds (13 against G. mellonella and 20 against scarabs) was 153- and 72-fold, respectively. Infective juveniles had difficulty locating diapausing P. japonica larvae, and selection did not result in any enhancement in finding these larvae. Selection for host-finding of scarabs resulted in a 24.6% gain in host-finding of G. mellonella , suggesting selection was for a general feature present at different levels in different hosts. The use of KOH to absorb CO2 resulted in the complete inhibition of S20 strain response to insect hosts, verifying that nematodes have been selected for enhanced sensitivity to CO2. S20 nematodes were significantly less storage stable than the Foundation strain.

Movement patterns of japanese beetle, Popillia japonica Newman, and European chafer, Rhizotrogus (Amphimallon) majalis (Razoumowsky), grubs as influenced by gravity, host plant position, and external disturbances were studied in laboratory soil–turfgrass microcosms. Second through third instars just before pupation were monitored using radiographic techniques. Neonates were monitored using destructive sampling. Results demonstrate significantly different movement patterns between species and among age groups. The development stage of the grub had a large effect on Japanese beetle grub behavior and a measurable, but lesser effect, on European chafers. All larval stages of European chafers and all larval stages of Japanese beetle, except neonates and postoverwintering third instars, displayed a downward movement in response to disturbance. Neonate Japanese beetles showed little movement while postoverwintering. Japanese beetles moved upward when disturbed. European chafer grubs of all age classes displayed random vertical movement with some arrestment in or near sod. Preoverwintering and postoverwintering third-instar chafers showed less dramatic arrestment behavior than other instars tested. Second-instar Japanese beetles behaved similarly to European chafers; however, third instars behaved very differently. All third instars except those tested in late winter and early spring showed some innate downward movement in the soil microcosms. Japanese beetles tested in late winter displayed random movement with some arrestment in sod, whereas those tested in early spring exhibited upward movement and arrestment in sod.

A novel cry gene, cry8Db, highly toxic to scarab beetles such as the Japanese beetle, Popillia japonica Newman, was cloned from an isolate of Bacillus thuringiensis(Bt), BBT2-5. The cry8Db gene has 3525bp nucleotides and codes for a protein of 1174 amino acid residues. The protein, Cry8Db, has typical Bt characteristics such as the 8-block, conserved sequences and the three-domain 3D toxin structure as defined with Cry3Aa. When the amino acid sequence of Cry8Db was compared with that of Cry8Da whose gene was cloned and characterized in our laboratory earlier, substantial sequence diversities were found in their Domain III. The cry8Db gene was expressed in an acrystalliferous B. thuringiensis strain, BT51. BT51 expressing cry8Db formed a spherical crystal like the natural crystal of BBT2-5. The Cry8Db protein was assayed along with the other scarab active Cry8Da and Cry8Ca against the Japanese beetle. While Cry8Da and Cry8Db had toxicity against both adults and larvae of the Japanese beetle, Cry8Ca was toxic to only larvae. Cry8Ca showed no toxicity against the adult beetle up to 30 microg per 1 cm(2) of leaf discs on which the protein was applied. The activation process of Cry8Db by adult and larval gut juice was compared in vitro with the processes of Cry8Da and Cry8Ca. All three proteins, Cry8Db, Cry8Da and Cry8Ca, produced a toxic core of approximately 70kDa equally indicating that the activation process does not inactivate the adult activity of Cry8Ca. We concluded that the adult activity of Cry8D proteins is encoded in Domain II. Further tests against other beetle species showed a significant difference between Cry8D's and Cry8Ca but no difference between Cry8Da and Cry8Db. Comparison of 3D structural models of Cry8Ca, Cry8Da and Cry8Db, which were constructed by using Cry3Bb as the structural template, indicated significant structural differences, especially between Cry8Ca and Cry8D proteins, in three major surface-exposed loops of Domain II that may be involved in determining the adult beetle activity.

The possible effect of soil pH on Japanese beetle, Popillia japonica Newman, oviposition and early-instar survival was investigated in the laboratory. Soils were amended with elemental sulfur or dolomitic limestone to produce a range of pH levels from 3.2 to 7.9. Pots containing one of seven soil pH levels that had been planted with a turfgrass mix were placed in cages and exposed to Japanese beetle adults for 7 or 10 d. Pots were subsequently inspected for eggs and first instars. In six tests conducted over two summers, there was no significant difference in Japanese beetle oviposition preference in pots maintained at pH 5.0-7.9.

Japanese beetle, Popillia japonica Newman, population densities and prevalence of the pathogens Ovavesicula popilliae Andreadis & Hanula and Bacillus popilliae Dutky are reported from three locations in Connecticut over a 3-yr period. Populations decreased to very low levels ( 50%). Conversely, prevalence of B. popilliae was

Total oxygen consumption ($V_{O_{2}}$ averaging 4.66 mL/h) of Japanese beetles (Popillia japonica Newman, Scarabaeidae, Coleoptera) during free flight was independent of body mass, while the mass-specific oxygen consumption $\dot{V}_{O_{2}}$ (averaging 52.14 mL/g · h⁻¹) was inversely related to body mass. Variations in wing beat frequency, ambient temperature ($T_{a}$), and thoracic temperature ($T_{th}$) did not alter $\dot{V}_{O_{2}}$. However, during the first few seconds following takeoff, flight metabolism was substantially increased. No change in wing beat frequency occurred during this time period; however, in separate experiments, larger wing beat amplitudes were observed during initial flight. Correspondingly, thermoregulation was more pronounced during takeoff and gradually disappeared in prolonged flight. Measured $T_{th}$ excesses were supported by data on metabolic heat gain and heat loss based on cooling curves. In resting beetles, whole-animal $\dot{V}O_{2}$ (0. 16 mL/h, averaged over a ran...

Marked Japanese beetles, Popillia japonica Newman, were released and recaptured in experimental plots to test the effect of contrasting types of strip-cropped soybean agroecosystems on beetle dispersal. Experimental treatments were soybean monoculture, soybean strip-cropped with dwarf sorghum (≍1 m tall), and soybean strip-cropped with tall sorghum (>2 m tall). Three hundred marked Japanese beetles were released in the center soybean strip of each treatment in small (0.11 ha) and large (0.45 ha) experimental plots. Beetles remained longer in the center soybean strip in the strip-cropped bicultures than in the monocultures indicating that the sorghum strips inhibited their movement. Dispersal rates were similar in dwarf and tall sorghum treatments despite differences in plant height. Significantly more marked beetles in the small plots, however, emigrated from the ends of the center soybean strip in the tall sorghum biculture than in the other two treatments indicating that tall sorghum oriented beetles to move parallel to intercropped strips. Japanese beetles responded to the sharp boundaries between host and nonhost habitat patches, as has been previously reported for host specific herbivorous insects. Results suggest that strip-cropping can affect the movement of polyphagous herbivorous insects and provide benefits for pest management in large-scale agroecosystems.

The host response to infection and tissue susceptibility of larval and adult Japanese beetles,Popillia japonica Newman, to the microsporidium,Ovavesicula popilliae Andreadis & Hanula, are reported. The normally transparent Malpighian tubules of Japanese beetle larvae, were hypertrophied and white in color when infected withO. popilliae, a microsporidian which also infects larval fat body, epidermis and pericardial cells. In addition to these tissues, œnocytes and tracheal epithelial cells were also infected in adults. Adult and larval reactions to infection included hypertrophied cells and melanization of the pericardium, but only larvae exhibited an intense inflammatory response. The discoloration of the pericardium most likely resulted from an accumulation of melanin.