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We studied ant predation on eggs and larvae of the black cutworm, Agrotis ipsilon (Hufnagel), and on eggs of the Japanese beetle, Popillia japonica Newman, on golf courses and lawns in central Kentucky. Lasius neoniger Emery accounted for >99% of the ant mounds on golf putting greens and collars. Although often regarded as a nuisance pest, L. neoniger preyed heavily upon A. ipsilon eggs on turfgrass cores implanted into putting greens, collars, fairways, and roughs. Predation on eggs was lower in fairways than in roughs, and in plots where ant populations were reduced by insecticides. When 1,600 individual 1st-instar cutworms were placed near L. neoniger nests on putting greens, 62% were attacked and killed upon their 1st encounter with the ants. Third- and 4th-instar cutworms generally fended off attacks by L. neoniger and Formica pallidifulva nitiventris Emery, but were invariably killed during encounters with Formica schaufussi Mayr and Formica subsericea Say, larger ants that are common in lawns and golf roughs. Predation on implanted Japanese beetle eggs also tended to be greater in roughs than in fairways, and fewer grubs were found in areas of golf roughs where ants were abundant than where ants had been controlled. This study suggests that predation by indigenous ants provides an important buffer against pest outbreaks on lawns and golf courses.

We tested biological control agents for the control of 3rd-instar scarab turfgrass pests, both for the masked chafer Cyclocephala hirta LeConte and the Japanese beetle, Popillia japonica Newman. The former species is endemic in California whereas the latter, although not yet established, constitutes a permanent serious threat to agriculture and horticulture in California. We conducted experiments using C. hirta in California and P. japonica in New Jersey. A field trial conducted in 2 different California turfgrass sites compared the field persistence in the absence of hosts of Bacillus thuringiensis Berliner subspecies japonensis Buibui strain, the milky disease bacterium, Paenibacillus (=Bacillus) popilliae (Dutky), and the entomopathogenic nematodes Steinernema kushidai Mamiya and Heterorhabditis bacteriophora Poinar to that of the organophosphate diazinon. Soil samples taken 0-70 d after applications were bio-assayed with P. japonica. Only diazinon and the entomopathogenic nematode S. kushidai caused substantial mortality and S. kushidai activity persisted significantly longer than diazinon activity. In greenhouse experiments, combinations of entomopathogenic nematode species usually resulted in additive mortality of scarab larvae. Combinations of S. kushidai and diazinon also resulted in additive mortality. In field trials, the efficacy of H. bacteriophora and especially S. kushidai and S. glaseri, was comparable to that of diazinon over 14-18 d. However, it is likely that at least S. kushidai would have outperformed diazinon over an extended period because of its longer persistence and potential for recycling in the hosts. S. kushidai, should it become commercially available, deserves further examination as an alternative to chemical white grub control especially as a highly compatible component of sustainable turfgrass management.

Root-feeding white grubs (Coleoptera: Scara baeidae) are among the most important group of pests attacking grasses in pastures and land scapes in the United States. Throughout the con tinental United States, about 10 species of grubs and several species in the genus Phyllophaga are reported pests of turfgrass (Vittum et al. 1999). In southeastern states, grubs of Phyllophaga, south ern masked chafer (Cyclocephala lurida), the green June beetle (Cotinus nitidia), and Japanese beetles (Popillia japonica) are traditionally asso ciated with damaged turfgrass. However, recent publications (Buss 2006, 2008) challenge the as sumption that just a few species account for the alleged increase in damage to turfgrass from white grubs in the southeast. Furthermore, intro duced and native species may be expanding their range into or throughout the southeast. This pa per documents the occurrence of white grubs of the Asiatic garden beetle Maladera castanea (Ar row), an exotic species, in damaged turfgrass in Alabama. Asiatic garden beetles, native to China and Ja pan, were first detected in New Jersey in 1921. They have since spread to most of the northeast ern states, west to Indiana, and south to South Carolina (Vittum et al. 1999, NAPIS 2008). Spread of Asiatic garden beetles in the northeast was attributed to natural dispersal of adults as well as movement of infested nursery stock and turfgrass (Hawley & Hallock 1936). All known biological information is based on only a few reports (Hawley 1931; Hawley & Hal lock 1936) of populations in the northeastern United States. Adults are nocturnal, emerging when temperatures are 18.3-21.1?C (65-70?F), and actively fly at temperatures >21.1?C (Hawley & Hallock 1936). Adult flight begins in late Jun and can last into the fall. At night, adults are at tracted to lights and light traps are the suggested means of trapping adults (Hawley & Hallock 1936). Adults feed on foliage or flowers of 100 or more hosts (Hawley & Hallock 1936), and spar ingly on grass blades (Heller 1995). Damage to woody and herbaceous plants from adults can be misdiagnosed since the damage appears over night and adults are rarely observed. Damage to foliage begins from the edges, progressing inward until only the midvein remains. Adults seem to prefer flowers attacking roses, chrysanthemums, dahlia, sunflower, strawflowers, and zinnias (Hawley & Hallock 1936), aster, and goldenrod (Vittum et al. 1999). Even though adult damage can be extensive when present, adults are only considered minor pests of ornamentals (Vittum et al. 1999). In daylight hours, adults are in the soil near host plants or in turf adjacent to these host plants. Females lay eggs in clusters in the soil and seem to prefer weedy patches of turf that are well irrigated (Hawley & Hallock 1936; Heller 1995). Eggs are present in the soil beginning in Jul. Eggs absorb moisture from the soil, swell, and hatch (Hawley & Hallock 1936) similarly to other white grubs. Newly-emerged grubs are 1.4 mm long, and develop to 19 mm when mature. There are 3 instars with mature grubs present in the mid-Oct. Grubs can be found under weedy turf, or in fields of sweetpotato, beans, and other field crops (Haw ley & Hallock 1936). Grubs are distinguished from other scarabs by a distinctive raster pattern and enlarged bulbous stipes on the maxillae (Hawley & Hallock 1936; Ritcher 1966). In the northeastern United States, there is 1 generation per year. On 2-3 Oct, 2008, a stand of weedy turfgrass on the Turfgrass Research Unit in Auburn, AL was sampled for grubs. This stand, 27.2 x 21.9 m (90 x 72 ft), is composed mostly of crabgrass (Digitaria sp.), dichondra, and various sedges and grasses. This stand was established in 1997 on a sandy loam soil and recently has been used for herbicide research, and in 2008, the area was irrigated weekly with about 2.54 cm (1 inch) of supplemen tal water and maintained at 3.8 cm (1.5 inch). The area generally stays moist after rain and wet dur ing the winter months. The grubs collected on both days were mostly southern masked chafer (Cyclocephala lurida), a few billbug larvae, 1 Japanese beetle (Popillia japonica) grub, and 5 unidentified larvae. All grubs were collected into containers along with field soil and transported to the lab. The previ ously unidentified grubs had a raster pattern and swollen stipes characteristic of Asiatic garden beetle. The grubs were preserved in ethanol and the identity later confirmed with the key of Ritcher (1966). Grubs had an average length of 17.1 mm (0.67 inch) and head capsule width of 2.77 mm. These data indicate that the grubs are likely third instars (Hawley & Hallock 1936; Ritcher 1966). This is the southernmost report of Asiatic gar den beetle in the United States. Recent trapping studies in Alabama (Flanders et al. 2000) and, in Florida, Buss (2006) did not detect adult M. casta nea in blaeklight trap samples. The Alabama study, conducted in pastures, was within 50 km of

Nine commercial plant protectants were tested for efficacy against feeding by adult Japanese beetles (Popillia japonica Newman). Treatments included an endotoxin from a bacterium [Bacillus thuringiensis (Berl.) var. san diego]; microencapsulated pyrethrum extracted from pyrethrum [Tanacetum cinnerariifolium (Trev.) Schultz-Bip.], two extracts from neem (Azadirachta indica A. Juss); an extract from cayenne pepper (Capsicum annuum L. var. annuum Longum Group); an extract from garlic (Allium sativum L.); rotenone extracted from galedupa (Derris trifoliata Lour.) and barbasco [Lonchocarpus sericeus (Poiret) Kunth] or timbo (L. nicou Aublet D.C.); carbaryl (1-napthyl methylcarbamate); and the pyrethroid, fenpropathrin. Experimental plots were located at the Horticulture Field Laboratory (HFL), Raleigh, and the Mountain Horticultural Crops Research and Extension Center (MHCREC), Fletcher, NC. Himalayan birches [Betula utilis var. jacquemontii (Spach) Winkl.] were used as host plants. Treatments were applied twice, 2 weeks apart. Five weeks after initial application, trees treated with fenpropathrin averaged 2% defoliation vs. 40% defoliation for the control trees at HFL; and 3% defoliation vs. 100% defoliation for the control trees at MHCREC. Rotenone treatments averaged 10% defoliation at HFL and 92% defoliation at MHCREC. The following treatments were not significantly different from the control at week 5 at either location: garlic extract, neem extracts, cayenne pepper extract, microencapsulated pyrethrum, encapsulated bacterial endotoxin, and carbaryl.

Imidacloprid, a chloronicotinyl, and halofenozide, a bisacylhydrazine ecdysteroid agonist, recently have become widely used for residual control of scarabaeid grubs in turf. We evaluated their impact on earthworms and beneficial arthropods in field trials, and tested whether application in late spring might interfere with subsequent predation on black cutworm, Agrotis ipsilon (Hufnagel), and Japanese beetle, Popillia japonica Newman, life stages in Kentucky bluegrass, Poa pratensis L., turf. Bendiocarb, a short-residual carbamate, was included for comparison. Imidacloprid caused some short-term suppression of earthworms, whereas bendiocarb had severe impact on earthworms, mesostigmatid mites, and Collembola. Pitfall trap captures of predatory coleopteran larvae and hister beetles were reduced by imidacloprid and bendiocarb, but abundance of ants, carabids, spiders, and staphylinids was largely unaffected. Halofenozide caused no reduction in abundance of any group of beneficial invertebrates. Scavenging on fresh-frozen A. ipsilon larvae was reduced for ≈1 wk after use of imidacloprid or bendiocarb, but predation rates on eggs or pupae of A. ipsilon, and on implanted P. japonica eggs, were unaffected. This work suggests that application of halofenozide or imidacloprid, followed by irrigation, will have relatively little impact on beneficial invertebrates, although both compounds are persistent enough to control P. japonica and Cyclocephala spp. grubs eclosing several months later.

Autodissemination may be effective against the Japanese beetle, Popillia japonica Newman, in situations where habitats of its larvae are inaccessible. Trapping systems with attractants for both male and female Japanese beetles are commercially available. We fabricated an inoculation chamber which fits between the top of a standard Trece Catch CanTM Japanese beetle Trap and its holding canister. Beetles which are attracted to the trap fall through a hole in the inoculation chamber and land on a mesh screen. A partial funnel and canister attachment from a metal Ellisco Japanese Beetle Trap was secured beneath a hole in the floor at the opposite end of the chamber. A 10-cm section in the middle of the box, between the entrance hole in the roof and the exit hole in the floor, allows space for a dish containing the inoculum to be placed into the chamber through a door in the side of the unit. The trap has been tested with Metarhizium anisopliae (Metschnikoff) Sorokin as the pathogen. Beetles emerging from the ...

Two morphological types of Bacillus popilliae, causal agent of the milky disease, have been isolated from taro beetles (Papuana spp, Coleoptera: Scarabaeidae). B. popilliae from P. woodlarkiana woodlarkiana (Papua New Guinea) was a type A1 with a small sporangium (4.1 x 1.6 microm) and a large spore (2.1 x 1.4 microm) and parasporal body (1.8 x 1.2 microm) that sometimes overlap. B. popilliae from P. uninodis and P. woodlarkiana laevipennis (Solomon Islands) was a type B2 with a small sporangium (2.8 x 1.3 microm), a small eccentric spore (1.1 x 0.7 microm), and no parasporal body. The infectivity of these B. popilliae to Papuana uninodis larvae was compared with two B. popilliae samples from Popillia japonica in injection tests. The hemolymph of P. uninodis supported the germination and growth of isolates from Papuana and P. japonica. Results were similar in third instars and adults. Highest infection (spores present) and mortality was caused by the isolates from Papuana: mortality reached almost 100% 4 weeks after injection of the B2 type B. popilliae with 40% of larvae and 52% of adults infected. Injection of type A1 caused lower mortality but a similar percentage infected. Of two A1 B. popilliae from P. japonica, one caused a mortality comparable to type A1 from Papuana but lower infection; an older isolate resulted in low mortality and only one infected larva. B. popilliae type A1 from P. woodlarkiana was produced in the Solomon Islands by injection of spores in P. uninodis. Thirty four percent of the injected larvae and 31% of the adults produced spores with an average yield of 3.2 and 0.8 x 10(9) spores/insect, respectively. Oral application of a single dose of 10(7) spores of the B. popilliae isolates from P. uninodis or P. japonica did not cause infection and similarly inoculation of the food with spores of B. popilliae type B2 did not result in infections. However, when different rates were applied to the food of second- and third-instar P. uninodis, the B. popilliae type A1 from P. woodlarkiana caused up to 15% infection and concentration-related mortality.

Selected elm, Ulmus spp., and their hybrids growing at The Morton Arboretum, Lisle, IL, were evaluated in bioassays for feeding preference by the Japanese beetle, Popillia japonica Newman. No-choice bioassays revealed that the hybrids U. ‘Morton’-Accolade ( U. japonica x U. wilsoniana ), U. elongata, U. laciniata, U. lamellosa, U. macrocarpa, U. parvifolia, U. ‘Morton Plainsman’-Vanguard ( U. japonica x U. pumila ) x U. davidiana, and U. Morton Plainsman-Vanguard ( U. japonica x U. pumila ) x U. japonica-wilsoniana-pumila had low ( 68%) amounts of leaf tissue consumed. Rankings for the dried fecal pellet weights, field defoliation survey, and multiple-choice studies were consistent with the no-choice studies for these same elm genotypes. Leaf pubescence on U. glaucescens, U. lamellosa, and U. macrocarpa seems to be a feeding deterrent for the adult beetle as evidenced by low levels of feeding damage (

It is commonly held that food-aversion learning should be more prevalent in polyphagous herbivores than in specialists. We tested the ability of Popillia japonica, a polyphagous scarab, to learn avoidance of a palatable but illness-inducing plant. Beetles that feed on flowers of geranium, Pelargonium × hortorum, became paralyzed, although most recovered within 24 h. In choice tests, naive beetles strongly preferred geranium petals over leaves of linden, Tilia cordata, a highly suitable host. Experienced beetles maintained this preference although it resulted in repeated bouts of paralysis. Fecundity was >10 times higher for beetles fed linden foliage for 2 wk than for those fed only geranium. Nevertheless, when a surplus of both foods was provided, the beetles fed mainly on geranium, resulting in greatly reduced fecundity. These results contradict the view that generalists should show propensity for food-aversion learning. Indeed, in this case, P. japonica continued to prefer the toxic plant, compromising its fitness.

Field populations of Japanese beetle, Popillia japonica Newman; European chafer, Rhizotrogus ( Amphimallon ) majalis (Razoumowsky); oriental beetle, Exomala orientalis (Waterhouse); and Asiatic garden beetle, Maladera castanea (Arrow), were exposed to the ecdysone agonist halofenozide in turf plots. In mixed-species larval populations, a higher dosage of halofenozide (1.7–2.2 kg [AI]/ha) was required to give significant suppression of European chafer than was required to control Japanese and oriental beetles (1.1 kg [AI]/ha). In other experiments, halofenozide (1.1–1.7 kg [AI]/ha) reduced Japanese beetle and oriental beetle populations by 75–98%. These data confirm predictions of selectivity based on previous laboratory studies. Asiatic garden beetle survival in the field was 40–50% higher in plots treated with halofenozide than in untreated plots. Dosage effects were compared for European chafer and Asiatic garden beetle 3rd instars in the laboratory; Asiatic garden beetles were insensitive to halofenozide at all tested dosages. Asiatic garden beetle and European chafer may increase in relative importance where halofenozide is used against mixed populations of exotic scarabs.

The host immune response is a key obstacle to entomopathogenic nematodes in making the transition from the free-living state to parasitism. The entomopathogenic nematode Steinernema glaseri has evolved mechanisms to evade immune encapsulation in larvae of the Japanese beetle, Popillia japonica. Host intrahemocoelic injection tests show that live axenic nematodes of S. glaseri not only avoid host melanotic encapsulation but also protect dead nematodes injected after the live ones. This result indicates that the nematodes release anti-immune factor(s). We extracted the nematode surface coat proteins and found that at least one protein (SCP3a) from the S. glaseri surface coat can suppress the host immune system. This suppression protects unrelated nematode species from encapsulation and latex beads from phagocytosis. We conclude that S. glaseri uses an anti-immune protein to defeat the host immune system, thereby protecting itself from encapsulation. Presumably its symbiotic bacteria are similarly protected from phagocytosis.

The basis for resistance of Bradford callery pear, Pyrus calleryana Decaisne ‘Bradford,’ [Rosaceae] to the Japanese beetle, Popillia japonica Newman, was investigated. Chloroform-dipping rendered leaves palatable, initially suggesting that deterrent waxes had been removed. However, extracts containing surface waxes were not deterrent. Subsequent experiments showed that increased palatability of solvent-dipped leaves is associated with enzymatic tissue browning, characteristic of polyphenol oxidases, rather than simply release of phagostimulants from surface disruption of damaged leaves. Frozen and thawed leaves showed similar browning, becoming increasingly palatable for several hours after thawing. Palatability changes were temperature- and aerobic-sensitive, further evidence that oxidizing enzymes are involved. Juice from leaves that had been frozen and thawed stimulated feeding on glass fiber disks, whereas fresh leaf juice did not. Survival and fecundity were much higher for beetles fed frozen and thawed or chloroform-dipped Bradford pear leaves than for beetles fed normal leaves. We hypothesize that decompartmentalization of deterrent compounds, possibly phenolics, followed by enzymatic oxidation and altered leaf chemistry may explain the increased palatability of chloroform-dipped or frozen and thawed Bradford pear tissue to P. japonica. This approach may be helpful in identifying specific compounds responsible for resistance of woody plants to generalist insects.

Attraction of Japanese beetles, Popillia japonica Newman, to 17 compounds typical of those released by undamaged and insect-damaged angiosperm leaves was compared in replicated field trials. Eight compounds attracted significantly more beetles than did an unbaited trap; however, the 2 most attractive single compounds, phenylacetonitrile and ( Z )-jasmone, were only ≍10% as effective as geraniol, a component of commercial lures. Progressive pairwise addition of less attractive volatiles to a blend of phenylacetonitrile and ( Z )-jasmone resulted in increasing beetle capture. The relative attractiveness of blends of conifer/eucalyptus, fruit-like, and floral odors, plus a complete mixture of these, also was tested. Although the floral, fruit-like, and complete mixtures captured more beetles than did the conifer/eucalyptus blend or phenylacetonitrile alone, our results suggest that this polyphagous insect is likely to be attracted to many naturally occurring plant volatiles, and that as the number of components in a volatile blend increases, so will its attractiveness. The complete mixture of volatile compounds also attracted more green June beetles, Cotinis nitida (L)., than did the fruit-like, floral, or conifer/eucalyptus blends.

An extract of adult Japanese beetles, Popillia japonica Newman (Coleoptera: Scarabaeidae), collected in commercial Japanese beetle traps, afforded a large amount (nearly 0.2 mg/g of fresh weight) of cholesterol. As phytophagous insects, Japanese beetles feed on a variety of plants. Since little or no cholesterol can be obtained from their diet, they must be able to utilize phytosterols to generate and store cholesterol.

Despite the remarkable diversity of the sex pheromone chemistry in scarab beetles, various species utilize a common type of γ-lactones in their chemical communication channels. These compounds differ primarily in length of the alkenyl side chain and the stereochemistry at the chiral center. Two species, Anomala osakana and Popillia japonica , utilize the opposite enantiomers of japonilure as sex pheromones. Each species produces only one of the enantiomers that functions as its own sex pheromone and as a behavioral antagonist to the alloreceiver. Pheromone binding proteins (PBPs) have been characterized, which are present in these and several other scarab species. In most cases there was only one class of PBP, which was expressed in both sexes. A. osakana and P. japonica possess each one single PBP with high homology to each other. In each species the same PBP seems to recognize both enantiomers of japonilure, i. e., the pheromonal and the “stop” signals. Based on the N-terminal sequences, the antennae-specific proteins from various other species were highly conserved within the family and showed moderate homology to putative odorant binding protein from Drosophila melanogaster (47%), Lygus lineolaris (45∼50%) and the ABPX protein from Bombyx mori (30∼35%). From analysis of extracts of soluble antennal proteins from several species, significant degradation of the γ-lactones (buibuilactone, japonilure) was detected, essentially in all of them, even in species that do not use these compounds as pheromones. Recently a peculiar pheromone with a diamide moiety [1, 3-dimethyl-2, 4-(1 H , 3 H )-quinazolinedione] was isolated from Phyllopertha diversa , which was rapidly degraded by antennal enzymes from this species. Beetles that utilize lactones as their pheromones possess little or no ability to metabolize this compound.

Entomopathogenic nematodes (Rhabdita: Heterorhabditidae and Steinernematidae) have been effective as inundative biological control agents of scarab larvae (Coleoptera: Scarabaeidae) in turfgrass. Entomopathogenic nematodes also occur naturally in turfgrass and endemic or inoculated populations may be able to provide effective long-term control. Variation in Heterorhabditis bacteriophora and Steinernema carpocapsae spatial and temporal distribution along transects placed at different turfgrass sites in central New Jersey, USA, was investigated. H. bacteriophora tended to be recovered from fewer sections in a transect than S. carpocapsae, but the two species, overall, did not differ in patchiness of distribution. In one transect with a H. bacteriophora population S. feltiae was also recovered, but the two populations seldom overlapped spatially. In transects with adequate scarab larvae density for analysis, H. bacteriophora density and Popillia japonica larvae density were inversely correlated. This suggests that endemic H. bacteriophora populations may suppress P. japonica populations. In one transect, an epizootic of H. bacteriophora in an undetermined host may have occurred. Edaphic factors were relatively uniform along transects and were, at most, weakly correlated with nematode recovery. Uniform inoculative releases of H. bacteriophora tended to return to patterns of distribution typical of endemic populations.

Endogenous allelochemicals can have a profound influence on host plant resistance to arthropod pests. This study evaluated 22 naturally-occurring compounds found in Rosaceous trees for their influence on feeding of adult Japanese beetles [Popillia japonica (Newman)]. Individual compounds were incorporated into artificial diets at concentrations from 0 to 100 mM. No-choice feeding trials were conducted over 24 hours. Four general trends were recognized among the dose responses: 1) no response: benzaldehyde, calcium oxalate, tannic acid, 2) stimulatory: rutin, 3) optimal peak: benzoic acid, phloridzin, quercetin, catechin, geraniol, arbutin, caffeic acid, chlorogenic acid, and 4) inhibitory: p-coumaric acid, eugenol, amygdalin, phloretin, naringenin, o-coumaric acid, arginine, asparagine, oxalic acid, and gallic acid. These results suggest that variation in chemical constituents and concentrations may have a strong influence on host plant resistance to Japanese beetles among Rosaceous plants.

The volatile compounds liberated by Japanese beetle ( Popillia japonica Newman) feeding on grape leaves ( Vitis labrusca L.) were studied. Ten consecutive collections of 3 hr duration were performed on live vines starting at 09:00 and continuing until 15:00 the following day. Release of most compounds followed a diurnal pattern, with the period of peak emission from 12:00 to 15:00 and the period of least emission from 00:00 to 03:00. Nineteen compounds were identified from the beetle-damaged vines, most of which were aliphatic aldehydes, alcohols and esters as well as terpene hydrocarbons. During periods of peak emission, volatile production from beetle-damaged vines was about 50-times higher than that of undamaged vines. © 1997 Elsevier Science Ltd. All rights reserved

We examined the preferences and performances of five beetle species (four chrysomelids and one scarab) on two species of willows (Salix sericea and S. eriocephala) and their interspecific hybrids. Beetle species differed markedly in their responses. In preference assays, two chrysomelid beetle species (Calligrapha multipunctata bigsbyana and Plagiodera versicolora) preferred hybrids, two chrysomelids (Chrysomela scriptaand Ch. Knabi) preferred hybrids and S. sericea, and the scarab beetle (Popillia japonica) preferred S. eriocephala. Experiments with purified salicortin indicated that salicortin concentration may contribute to these preferences. The relative performance (growth rate, pupal/adult weight and survivorship) of these beetles on the three willow taxa did not correspond with their feeding preferences. Three species exhibited intermediate performance on hybrid willows (the two Chrysomelaspp. and P. japonica); the Chrysomelaspp. performed best on S. sericea, while P. japonicaperformed best on S. eriocephala. One species performed equally well on all three taxa (C. multipunctata bigsbyana). The performance of Pl. versicolora was not tested. Our results support the general pattern that willow taxa with phenolic glycosides are more acceptable to specialist willow herbivores while those taxa without phenolic glycosides are more acceptable to generalist herbivores. We also show that to predict the relative susceptibility of hybrid and parental plants to herbivores, consideration must be given to the inheritance of traits affecting both preference and performance.

Cucurbitacins, the bitter triterpenes common to all Cucurbitaceae, are thought to be potent feeding deterrents for all insects not adapted to exploiting cucurbits. Yet appropriate choice tests not confounded by possible effects from phytochemical induction have only been conducted on a few chrysomelid leaf beetle spp. To examine the extent to which cucurbitacins deter feeding across several taxa of mandibulate and haustellate insects, we conducted feeding preference tests, pitting host tissues exogenously coated with a methanol dilution of cucurbitacin B against tissues coated with only methanol. Four mandibulate herbivores [ Popillia japonica Newman, Cerotoma trifurcata (Forster), Leptinotarsa decemlineata (Say) and Trichoplusia ni (Hubner)], 2 mandibulate detritivores [ Tenebrio molitor L. and Nauphoeta cinerea (Olivier)] and 4 haustellate herbivores [ Gargaphia solani Heidemann, Corythucha ciliata (Say), Peregrinus maidis (Ashmead), and Acyrthosiphon pisum (Harris)] were examined. We also investigated the ability of cucurbitacin B to deter Ostrinia nubilalis (Hubner) and Spodoptera exiqua (Hubner) females from ovipositing on treated substrates. Two patterns emerged from our feeding assays: nonadapted insects with mandibulate mouthparts were deterred from feeding on food tainted with cucurbitacins, whereas insects with haustellate mouthparts preferred cucurbitacin-treated food. Oviposition assays with O. nubilalis and S. exiqua revealed significant deterrence at doses of 15 μ g cucurbitacin B per square centimeter. The data suggest that cucurbitacins are generalized antifeedants for non-adapted mandibulate insects but may actually be phagostimulants for phytophagous insects with haustellate mouthparts. We report on the oviposition deterrence for this class of compounds.

The sex pheromone glands of female Anomala albopilosa albopilosa have been localized by extracting various parts of the body and analyzing the extracts by GC-MS and by histological and morphological studies. Female-specific epithelial cells line the inner surfaces of anal plates and two apical sternites; these cells are connected through many pores to the cuticle surface. The sex pheromones of other rutelines, A. cuprea and Popillia japonica and a melolonthine species, Heptophylla picea, have been also detected in the extracts from the same abdominal parts. On the other hand, the source of Holotrichia parallela sex pheromone was confirmed to be a ball-shaped abdominal sac that is exposed during the calling behavior, and no trace of the semiochemicals has been found in the anal plates and two apical sternites.

We studied the close range olfactory response of Japanese beetles, Popillia japonica Newman, to constitutive or feeding-induced volatiles from leaves of 4 maple species differing in susceptibility to defoliation. In pitfall arenas, intact leaves of susceptible species, Acer palmatum Thunberg and A. platanoides L., were not significantly more attractive than those of resistant maples, A. rubrum L. and A. saccharinum L. Consistent with these results, the compounds emitted by intact maple leaves were generally similar across resistance groupings. For each maple species, leaves that had been damaged overnight by Japanese beetles attracted significantly more beetles than did undamaged leaves. Beetles did not discriminate between damaged leaves of susceptible and resistant maple species. Significant differences were noted, however, in the types of feeding-induced compounds emitted by susceptible and resistant maple leaves. Specifically, leaves of susceptible species emitted relatively higher amounts of esters with fruit-like odors and the floral fragrance compound linalool, whereas leaves of resistant maples emitted more terpene hydrocarbons. Possible roles of volatile compounds as attractants and feeding stimulants, or as feeding deterrents, for the Japanese beetle are discussed.

Plants emit a series of characteristic volatile blends when damaged by insect feeding. Phenylacetonitrile is one of the volatiles from the leaves of the giant knotweed, Fallopia sachalinensis, infested by the Japanese beetle, Popillia japonica, or treated with exogenous airborne methyl jasmonate (MeJA). We examined the precursor of the nitrile and its origin in this system. l-Phenylalanine was determined to be a precursor of the nitrile in F. sachalinensis leaves, and the phenylalanine was also induced by beetle feeding and MeJA treatment. We also found that exogenous MeJA enhanced the biosynthesis of several amino acids in F. sachalinensis leaves.

Japanese beetle, Popillia japonica Newman, European chafer, Rhizotrogus (Amphimallon) majalis (Razoumowsky), and oriental beetle, Anomala orientalis Waterhouse, were exposed to an ecdysone agonist, halofenozide, in the egg, 1st instar, and 3rd instar at 1.5, 3, 6, and 12 ppm, respectively, in soil. Species, stage, and physiological condition of larvae affected the response to halofenozide. Japanese beetle was the most sensitive, with maximal response of 1st instars at 3 ppm. First and 2nd instars were more susceptible than were 3rd instars. Sublethal exposure of 1st-instar European chafer and 3rd-instar oriental beetle resulted in dose-dependent accelerated development and successful molts. Field trials are needed in locations where there are mixed populations of scarabs to determine whether the species differences in susceptibility to halofenozide observed in this work will translate to increased relative importance of European chafer.

Evaluation of trapped beetle repellency, various trap heights, and a string pheromone dispenser were tested on 2 golf courses for capture of Japanese beetle, Popillia japonica Newman, adults. Beetle captures increased significantly when trapped beetles were removed daily from traps. No significant differences in captures were found among traps with 121-, 18.9-, or 7.6-liter collection containers, and there was no interaction between beetle removal and collection container size. Trece traps with funnel rims at a height of 13 cm above ground level captured significantly more beetles than traps with funnel rims at ground level or at 90 cm above ground level. Numbers of beetles collected in traps with a polyvinyl chloride coated string pheromone dispenser were similar to numbers captured in traps using a standard Trece rubber septum pheromone dispenser.

A novel mechanism of reciprocal behavioral agonist-antagonist activities of enantiomeric pheromones plays a pivotal role in overcoming the signal-to-noise problem derived from the use of a single-constituent pheromone system in scarab beetles. Female Anomala osakana produce (S, Z)-5-(+)-(1-decenyl)oxacyclopentan-2-one, which is highly attractive to males; the response is completely inhibited even by 5% of its antipode. These two enantiomers have reverse roles in the Popillia japonica sex pheromone system. Chiral GC-electroantennographic detector experiments suggest that A. osakana and P. japonica have both R and S receptors that are responsible for behavioral agonist and antagonist responses.

Arborvitae ( Deja occidentals ‘Pyramidal’), Colorado blue spruce ( Peace pungent ), taxes ( Taxus x media ‘Densiformis’), and dwarf burning bush ( Eponymous Alta ‘Compacts’) nursery fields in Ohio and Michigan were sampled for Japanese beetle, papilla japonica Newman, adults and larvae. Nursery fields in counties with the highest adult catches were the most likely to have larvae in the field. Nursery fields where

The hypothesis that feeding-induced plant volatiles are responsible for aggregation of Japanese beetles, Popillia japonica Newman, on leaves was tested using clonal grape plants, Vitis Labrusca L. The attractiveness of undamaged vines, undamaged vines with nonfeeding beetles, vines with fresh feeding damage, and vines with overnight feeding damage was compared in replicated field trials by placing the treatments in the field in the morning and counting the number of beetles that landed on the plants throughout the day. Many more beetles were recruited to vines with overnight feeding damage than to any of the other treatments, indicating attraction to feeding-induced volatiles. Furthermore, the low number of beetles attracted to vines with nonfeeding beetles or freshly damaged vines provided evidence against the existence of a putative aggregation pheromone. Subsequently, we examined the emission of volatiles from grape vines in situ. During the period of peak emission (1200–1500 hours), emission from vines with overnight feeding damage was almost 65 times higher than from undamaged vines. Aliphatic compounds and terpenoids were the major classes of compounds emitted by the beetle-damaged vines. The implications of feeding-induced volatiles in host-finding and mate location by this polyphagous insect are discussed.

The Japanese beetle, Popillia japonica Newman, an introduced polyphagous scarabaeid, is an important pest of woody landscape plants and fruit crops in the eastern United States. A striking aspect of the beetle’s behavior is its tendency to congregate on particular plants, often nearly defoliating them, while other suitable hosts in the vicinity are only lightly damaged (Fleming, 1972). Feeding almost always begins at the top of a plant, regardless of its height, and proceeds downward (Fleming, 1972).

The volatile compounds emitted by leaves of four crabapple cultivars susceptible to damage by Japanese beetles and four relatively resistant cultivars were examined. Twelve compounds, mostly terpene hydrocarbons, were identified from intact leaves. The terpenes (E)-β-ocimene, caryophylene, germacrene D and (E,E)-α-farnesene occurred in significantly higher levels in susceptible cultivars, whereas resistant cultivars produced greater amounts of (E)-4,8-dimethyl 1,3,7-nonatriene and linalool. The relative attractiveness of the cultivars as determined in a pitfall bioassay, however, was not related to their susceptibility to the Japanese beetle as previously determined by defoliation sustained in the field. The attractiveness of individual cultivars was found to be positiviely correlated with linalool as a percent of the total volatile blend emitted by leaves. This study and previous work suggest that variation in susceptibility of crabapple cultivars to defoliation by Japanese beetles is not due to the attractiveness of the individual cultivars but rather to nonvolatile components of susceptibility and/or resistance. A scenario for host location by the Japanese beetle is presented.

Cultural practices were manipulated before or during seasonal flights of Japanese beetle, Popillia japonica Newman, and masked chafers, Cyclocephala spp., to study effects on grub densities in tall fescue or Kentucky bluegrass turf. Masked chafer grubs were consistently smaller and less abundant in turf that had been treated with aluminum sulfate to reduce soil pH and in high-mown turf. High mowing or application of aluminum sulfate before beetle flights reduced total biomass of white grubs in tall fescue by as much as 55 and 77%, respectively. However, where spatial gradients in soil moisture occur, the positive response of grub populations to moisture may override effects of those treatments. Grub densities were not affected by spring applications of lime or urea or by aerification of plots before beetle flights. Plots that were irrigated during beetle flights incurred significantly higher densities of both P. japonica and Cyclocephala spp. grubs than did nonirrigated turf. The use of a 2,247- kg roller to compact the soil before beetle flights did not affect subsequent grub populations, and the roller was not effective for remedial control of 3rd instars in the fall. Application of organic fertilizers (composted cow manure or activated sewage sludge) resulted in significant increases in grubs of green June beetle, Cotinis nitida L., in 1 of 2 yr. Cultural practices may have general or species-specific effects on densities of white grubs. This study suggests that witllolding irrigation during peak flight of beetles, raising cutting height, and light application of aluminum sulfate in spring may help to reduce the severity of subsequent grub infestations.

Descriptions or redescriptions along with comments on the distribution and habits of six species in the genus Pachystethus are provided. Anomala nutans Bates and A. vidua crassesculpta Bates are transferred to the genus Pachystethus and P. matzapatlecus new species, P. ixtacomitanus new species and P. nectoctenus new species are described from Oaxaca,Chiapas, and Veracruz, Mexico, respectively. Lectotypes for Popillia sticticollis, Anomala nutans, and A. vidua crassesculpta are designated. Color photographs, illustrations of diagnostic characters, and a key to species are also included.

Understanding the ability of natural populations of predators, parasitoids, and pathogens to suppress prey populations is important in determining their potential as biological control agents. We measured the seasonal population dynamics of endemic entomopathogenic nematodes ( Steinernema carpocapsae and Heterorhabditis bacteriphora ) in turfgrass and their effect on populations of Japanese beetle, Popillia japonica, larvae and mobile arthropods associated with the soil surface and thatch layer. Both nematode species were recovered in central New Jersey in samples collected from April to December. S. carpocapsae tended to be more prevalent in the plots (32.6 and 37.1% of sections) than H. bacteriophora (8.3 and 0.3% of sections). However, when only the positive sections were analyzed H. bacteriophora had a higher density (63.6 ± 49.3 nematodes/cm 2 ) than S. carpocapsae (17.0 ± 4.9 nematodes/cm 2 ). Sections with H. bacteriophora had significantly lower P. japonica densities compared to sections without nematodes (58.5 ± 20.2 compared to 112.7 ± 9.2 larvae/m 2 ). S. carpocapsae did not have a measurable impact on P. japonica populations. Mobile surface arthropods, as measured by pitfall catches, were higher in H. bacteriophora positive sections than in sections without nematodes. This difference was due to higher catches of Araneae and Lithobiomorpha. Mobile surface arthropod populations did not differ between sections with and without S. carpocapsae . The scope of H. bacteriophora′s impact on host populations is likely to be limited because of the nematodes patchy distribution.

Root-feeding white grubs (Coleoptera : Scarabaeidae) are among the most injurious insect pests of cool-season turfgrasses ; however, little is known about the influence of common lawn weeds on white grub populations. Greenhouse studies were conducted to evaluate the suitability of six weed species and one turfgrass including white clover (Trifolium repens L.), red clover (T. pratense L.), large crabgrass [Digitaria sanguinalis (L.) Scop.], annual bluegrass (Poa annua L.), buckhorn plantain (Plantago lanceolata L.), dandelion (Taraxacum officinale Weber), and Kentucky bluegrass (Poa pratensis L.) as food for grubs of the Japanese beetle (Popillia japonica Newman) and southern masked chafer (Cyclocephala lurida Bland). Third instars of both species fed upon and reduced root biomass of all weeds tested, and in some cases, reduced foliage yields as well. Survival of both grub species was as high, or higher, on most weeds as on Kentucky bluegrass. Dandelion and white clover appear to be particularly poor hosts for P. japonica and C. lurida, respectively.

Steinernema glaseri Steiner were applied in autumn field trials against Japanese, Popillia japonica Newman, and oriental beetle, Exomala orientalis (Waterhouse), larvae in 1992, 1993, and 1994. Nematodes were used in conjunction with various irrigation regimes and surfactants. The greatest reduction in larvae occurred where S. glaseri was used at 19.7 x 10 9 /ha and irrigated daily with 3.2 mm of water for 14 d. In 19 of 24 treatments where rates of 1.24-24.7 x 10 9 /ha were used with and without irrigation, S. glaseri reduced larval densities below a threshold of 8/0.09 m 2 . The use of 2 surfactants provided no additional control. Soil temperature data for Rhode Island revealed applications should be made before 1 September to maximize efficacy.

Adult females of the southern masked chafer, Cyclocephala lurida Bland, produce a volatile sex pheromone that attracts conspecific males. Previously we documented that extracts of 3rd-instar Cyclocephala spp. grubs contain one or more compounds that attract males and stimulate copulatory attempts when these males arrive at the source of the odor. Attraction of males to larvae appears to be undocumented for any other insect species. We examined developmental expression of the production of this attractant within Cyclocephala spp., and evaluated hexane extracts of other species of scarabaeid grubs for attractiveness to C. lurida or conspecifics. We found that the attractant is not limited to the postoverwintering last instar, but rather is present in all larval stages and in male and female pupae. Extracts of Cyclocephala spp. grubs also attracted males of the northern masked chafer, Cyclocephala borealis Arrow. Hexane extracts of 3rd instars of Japanese beetles, Popillia japonica Newman, and green June beetles, Cotinis nitida L., were not attractive to male C. lurida , indicating that there is some phylogenetic specificity to this unusual production of a sex-specific attractant. Third instars of C. nitida do not attract conspecific males. Studies of male attraction to conspecific immature stages in Cyclocephala spp. and other Scarabaeidae may provide insights into the evolutionary origins of chemical communication systems.

Response of Kentucky bluegrass, Poa pratensis L.; tall fescue, Festuca arundinacea Schreber; hard fescue, F ovina L. var. duriuscula; perennial rye grass, Lolium perenne L.; and creeping bent grass, Agrostis palustris (Hudson), to herbivory by root-feeding by grubs of Japanese beetle, Popillia japonica Newman, and southern masked chafer, Cyclocephala lunda Bland, was evaluated in greenhouse trials. Potted turf grasses were infested with initial densities equivalent to 73 or 146 grubs per 0.1 m2 in spring and fall trials, and effects on plant growth and grub survival were determined. All turf grasses tolerated significant damage to roots without loss of foliage yield. In fact, feeding by grubs stimulated increased growth of foliage in some grasses. With comparable densities of grubs, loss of roots tended to be proportionately less in creeping bent grass than in other grass species. Survival of grubs was similar in all turf grasses, and between low and high grub densities. P. japonica caused greater loss of roots than C. lurida in the spring trial, but the reverse was true in the fall. Root loss per grub decreased with increasing larval density, suggesting food limitation even though root systems were not completely devoured. Implications of these findings for tolerance and recovery of grub-damaged turf grasses are discussed.

Defense mechanisms of Japanese beetle larvae, Popillia japonica, and their effectiveness against entomopathogenic nematodes were investigated in the laboratory. Behavioral grooming by P. japonica larvae was effective in reducing invasion of both Steinernema glaseri and Heterorhabditis bacteriophora. Forty percent of H. bacteriophora infective juveniles were inactivated or killed in the host gut fluid within 8 hr, whereas 11% of S. glaseri were killed. Host melanotic encapsulation, induced by injection of 20 H. bacteriophora infective juveniles, caused 75% nematode mortality. S. glaseri was less susceptible to the host immune response, which caused only 12% mortality of injected nematodes. In a sand column assay, S. glaseri was found in the host hemocoel 8 hr earlier than H. bacteriophora. However, Photorhabdus luminescens, the bacterium of H. bacteriophora, was detected 16 hr earlier in the host's hemolymph than S. glaseri's bacterium, Xenorhabdus poinarii. Despite the differences in effectiveness of host defenses, S. glaseri and H. bacteriophora caused similar host mortality in the sand column assay. We demonstrated that the two nematode species use different strategies for invasion of P. japonica larvae. S. glaseri, which tolerated P. japonica gut fluid and avoided the host immune response, has a high initial rate of penetration and survival; H. bacteriophora, which exhibited poor tolerance to the host gut fluid and induced a strong host immune response, releases its bacteria during or soon after penetration to kill the host.

We examined the influence of proteinase inhibitors on digestive enzymes and development of oriental beetle, Exomala orientalis Waterhouse, European chafer, Rhizotrogus majalis (Razoumowsky), Phyllophaga white grub, Phyllophaga anxia (LeConte), cranberry root grub, Lichnanthe vulpina (Hentz), Japanese beetle, Popillia japonica Newman, Asiatic garden beetle, Maladera castanea (Arrow) (Coleoptera: Scarabaeidae), and the black cutworm, Agrotis ipsilon (Rottemburg) (Lepidoptera: Noctuidae). We demonstrated that all species within our test group had alkaline midguts that contained proteinase activity that could be inhibited, in vitro with serine proteinase inhibitors. Our data suggests that host range may influence the susceptibility to non-host inhibitors. Chronic ingestion of the serine proteinase inhibitor, Kunitz-soybean trypsin inhibitor (STI), significantly reduced proteolytic activity in vivo in those species with relatively specialized feeding habits (i.e., cranberry root grub, Japanese beetle, Asiatic garden beetle, and black cutworm). Chronic ingestion of STI also resulted in reduced larval growth and delayed pupation for black cutworm, and elevated larval mortality for Japanese beetle. However, chronic ingestion of STI did not influence larval survival for those species with relatively generalized feeding habits (i.e., oriental beetle, European chafer). Based on these results, we propose mechanistically-based criteria for selecting proteinase inhibitors for phytochemical defense against herbivorous insects.

Flowering crabapples, Malus spp., are important small landscape trees in much of the United States and southern Canada. We evaluated 42 cultivars of Malus spp. for resistance to defoliation by Japanese beetles, Popillia japonica Newman, for 2 growing seasons and found marked and consistent variation in resistance. Defoliation caused by beetles ranged from 100 to

Root-feeding white grubs (Coleoptera: Scarabaeidae) are important pests of cool-season turfgrasses in the eastern USA. We studied effects of irrigation and N fertilization on Kentucky bluegrass (Poa pratensis L.), and tall fescue (Festuca arundinacea Schreb.) response to grub damage under greenhouse and field conditions. Potted turfgrasses, with or without Japanese beetle (Popillia japonica Newman) grubs (122/0.1 m 2 ), were maintained under two irrigation regimes, with or without supplemental N fertilizer (204 kg N h -1 ). Foliar and root yields, and grub survival were measured after 4 wk. In the field, sod was grown in rooting boxes, with or without spring fertilization (146 kg N ha -1 ) and fall irrigation (2.54 cm wk -1 ). Half of the plots were infested with southern masked chafer (Cyclocephala lurida Bland) grubs (66/0.1 m 2 ) in late August and effects on aesthetic damage, surface temperature, foliar yield, root strength, and grub survival were measured for 9 wk. Grubs severely damaged roots of both grasses regardless of management regime. Within regimes, however, this damage significantly reduced foliar yield only in turf that received both N and high irrigation. In the greenhouse, grub-damaged grasses that received remedial N, nevertheless, had higher foliar yield than did non-fertilized grasses without grubs. In rooting boxes, fall irrigation following spring fertilization enhanced visual quality of grub-infested turf and root strength of grub-damaged Kentucky bluegrass. Survival of grubs did not differ between grass species or fertilization regimes, but more P. japonica survived in pots receiving high irrigation. Irrigation during the fall feeding period, followed by remedial N fertilization, may promote tolerance and recovery of grub-damaged turfgrasses.

Field-collected adult Japanese beetles, Popillia japonica Newman, treated with conidia of Metarhizium anisopliae Sorokin were compared with nontreated beetles under held conditions on Terceira Island, Azores, Portugal, using mark-release-recapture techniques, Distance, direction, and day of capture were monitored with lure-baited Japanese beetle traps placed in four concentric circles 50 to 1000 m around the release point during July and August of 1990 and 1991. Significantly fewer treated beetles (2.5%) were recaptured than untreated individuals (5.2%). Approximately 70% of the recaptured beetles were caught within 50 m of the release point, but a small percentage (

Japanese beetle adults, Popillia japonica, can become infected with and disperse the entomopathogenic nematode, Steinernema glaseri, under laboratory and field conditions. After a 24-h exposure to 10 000 infective juveniles/20 adult beetles, 45% of the beetles died within 4 days post-treatment, but only 59% of these were infected with the nematode. Corresponding control mortality was 6.5%. An average of 238 infective juveniles were produced/beetle. Beetles exposed to 4000 and 10 000 infectives/10 adults carried with them an average of 17 and 59 infectives/adult on external body surfaces respectively. When beetles that had been exposed to 4000 infectives/20 adults were transferred to, and held in, cages containing soil for 2 weeks, up to 89% of the adults died, as did 74% of the P. japonica larvae that were subsequently placed in the cages. When adults that had been exposed to 50 000 infectives/250 beetles in moist sand for 16 h were released into screened cages in the field at soil temperatures of over 25...

We evaluated tritrophic level interactions among fungal endophytes (Acremonium spp.) of fescue grasses (Festuca spp.), the root-feeding Japanese beetle Popillia japonica Newman larvae, and the entomopathogenic nematode Heterorhabditis bacteriophora Poinar. Third-instar P. japonica larvae were introduced into pots containing endophyte-infected or endophyte-free plants of tall fescue Festuca arundinacea Schreber (cultivars Kentucky 31 and Georgia Jesup Improved) and the Chewings fescue Festuca rubra commutata Guad. (cultivars F-93 and Jamestown II). After two weeks, the surviving larvae were recovered, and their susceptibility to nematodes was evaluated in sand columns. Endophytes enhanced the rate of nematode-induced mortality in all cultivars except Georgia Jesup Improved, and increased the proportion of dead larvae with nematodes in all cultivars except Jamestown II. Endophytes in the cultivar Kentucky 31 were associated with improved nematode establishment in the larvae. No effect on nematode reproduction was found. Since endophytes produce biologically active alkaloids, we tested the effects of an ergot alkaloid, ergotamine tartrate, on the feeding behavior and weight of P. japonica larvae in agar medium. The alkaloid caused feeding deterrence, and reduced the consumption of medium by the larvae, resulting in weight loss. These larvae were more susceptible to H. bacteriophora than the untreated larvae. Unfed ‘starved’ larvae were more susceptible to nematodes than those fed on untreated agar. Our results support the hypothesis that endophyte-induced starvation of P. japonica would reduce larval vigor, and render them more susceptible to entomopathogenic nematodes.

Cry8Da from Bacillus thuringiensis galleriae SDS-502 has insecticidal activity against both the larvae and adult Japanese beetle (Popillia japonica Newman). The receptor determines the specificity of the insecticidal activity of Cry proteins and hence, in order to reveal the mode of action of Cry toxin, receptor identification is a necessary step. However, a receptor for Cry8-type toxin has not been identified in the Scarabaeidae family of insects. Therefore, we aimed to identify the receptor of Cry8Da toxin in adult P. japonica BBMV. A ligand blot showed the Cry8Da toxin only bound to a 150kDa protein in the BBMV of adult P. japonica. In order to identify the Cry8Da toxin binding protein, it was purified by column chromatography and three internal amino acid sequences were determined. Two of the three internal amino acid sequences shared homology with Coleopteran β-glucosidases. In addition, the fraction containing the Cry8Da toxin binding protein had β-glucosidase activity but no aminopeptidase N and alkaline phosphatase activity, both of which are commonly reported as receptors for Cry toxins in Lepidopteran and Dipteran insects. The β-glucosidase homologous genes could be amplified by PCR using degenerate oligonucleotide primers designed from a conserved sequence of Coleopteran β-glucosidases and an internal amino acid sequence of the Cry8Da toxin binding protein. Taken together, the β-glucosidase in adult P. japonica BBMV is the receptor for B. thuringiensis Cry8Da toxin.

The impact of pathogens and parasitoids on the recently established population of Popillia japonica Newman in northwest Arkansas has been unknown. In this study, we quantified the prevalence of natural enemies: Stictospora villani Hays, Ovavesicula popilliae Andreadis and Hanula, Paenibacillus spp. (Dingman), nematodes and parasitic Diptera and Hymenoptera in third instar and adult populations in 2010 and 2011. S. villani was found in 38.4% and 35.5% of larvae in 2010 and 2011, respectively. S. villani was not found in adult beetles. Paenibacillus bacteria were not found in either larvae or adults in either year. In 2010, the microsporidian O. popilliae was not found in larvae but was present in 0.2% of adults. In 2011, 2.6% of larvae were infected with O. popilliae, but the microsporidian was not found in adults. A previously unknown Adelina sp. was found infecting 0.4% of adult beetles in 2010 and 1.3% of larvae in 2011. Nematode infections were found in 1.8% of larvae and 0.1% of adults in 2010 and not found in either life stage in 2011. No parasitic Hymenoptera or Diptera were found in either year. Apparently, pathogens and parasitoids currently provide little control of the Japanese beetle population within northwest Arkansas.

Larvae of the Japanese beetle, Popillia japonica , respond to attack by infective juveniles of the entomopathogenic nematode Heterorhabditis bacteriophora with aggressive and evasive behaviors. Brushing with the legs and rubbing with the abrasive raster were key aggressive behaviors associated with nematode attack in petri dish assays. Remarkably, contact with even a single live nematode triggered behavioral countermeasures directed at eliminating the attacker. Unrestrained P. japonica free to display aggressive behaviors removed more than 60% of attacking nematodes from their cuticle and experienced significantly less parasitism than larvae restrained from displaying defensive behaviors (13% or less of the nematodes removed). Unrestrained larvae were also capable of killing 20.6 ± 4.7% of the attacking nematodes. Observations of aggressive behavior in soil assays supported our petri dish results. Moreover, larvae were observed to respond to the presence of nematodes with evasive behavior. These defense behaviors are so generalized it appears unlikely that they evolved as a specific scarab evolutionary response to selection pressure from entomopathogenic nematodes, but rather to any threat or irritant.