Long-term suppression of turfgrass insect pests with native persistent entomopathogenic nematodes.

Entomopathogenic nematodes (EPNs) can control several important turfgrass insect pests including white grubs, weevils, cutworms, and sod webworms. But most of the research has focused on inundative releases in a biopesticide strategy using EPN strains that may have lost some of their ability to persist effectively over years of lab maintenance and / or selection for virulence and efficient mass-production. Our study examined the potential of fresh field isolate mixes of endemic EPNs to provide multi-year suppression of turfgrass insect pests. In early June 2020, we applied isolate mixes from golf courses of the EPNs Steinernema carpocapsae, Heterorhabditis bacteriophora, and their combination to plots straddling fairway and rough on two golf courses in central New Jersey, USA. Populations of EPNs and insect pests were sampled on the fairway and rough side of the plots from just before EPN application until October 2022. EPN populations increased initially in plots treated with the respective species. Steinernema carpocapsae densities stayed high for most of the experiment. Heterorhabditis bacteriophora densities decreased after 6 months and stabilized at lower levels. Several insect pests were reduced across the entire experimental period. In the fairway, the combination treatment reduced annual bluegrass weevil larvae (59 % reduction) and adults (74 %); S. carpocapsae reduced only adults (42 %). White grubs were reduced by H. bacteriophora (67 %) and the combination (63 %). Black turfgrass ataenius adults were reduced in all EPN treatments (43-62 %) in rough and fairway. Sod webworm larvae were reduced by S. carpocapsae in the fairway (75 %) and the rough (100 %) and by H. bacteriophora in the rough (75 %). Cutworm larvae were reduced in the fairway by S. carpocapsae (88 %) and the combination (75 %). Overall, our observations suggest that inoculative applications of fresh field isolate mixes of endemic EPNs may be a feasible approach to long-term suppression of insect pests in turfgrass but may require periodic reapplications.

Optimizing Sampling Technique Parameters for Increased Precision and Practicality in Annual Bluegrass Weevil Population Monitoring.

The annual bluegrass weevil (ABW), Listronotus maculicollis (Kirby), a significant pest of short-mown turfgrass in eastern North America, has developed widespread insecticide resistance because of excessive synthetic insecticide use. The proper monitoring of this pest may reduce insecticide applications in time and space. This study evaluated three sampling methods (soap flushing, vacuuming, and mowing) in golf course greens and fairways for monitoring adult ABW. Soap flushing was the most efficient method, especially with an 0.8% solution in two portions of 500 mL, extracting over 75% of the adults, and the extraction efficiency was not affected by the temperature or time of day. Vacuuming was more effective for recovering adult ABWs on greens (4-29% extracted) than on fairways (2-4%) but was not affected by the time of day. The extraction of adult ABWs in mower clippings was significantly affected by mowing height (higher recovery from greens versus fairways), and the efficiency decreased with the temperature. Adding a brush to the mower increased adult removal (from 15% to 24%) in greens at higher temperatures (18-25 °C); 70% of adults recovered in the clippings were unharmed. Overall, our findings suggest that soap flushing should be the preferred method for monitoring adult ABWs, and vacuuming might be a viable alternative for greens.

Insecticidal Activity of a Petroleum-Derived Spray Oil and an Organosilicone Surfactant on Listronotus maculicollis (Kirby) Adults in Laboratory and Greenhouse Bioassays.

The annual bluegrass weevil (ABW), Listronotus maculicollis (Kirby), is a severe pest of golf course turf in eastern North America. The development of pyrethroid- and multiple-resistant populations has created a dire need for novel tactics to control adults. We examined the insecticidal properties of a petroleum-derived spray oil (PDSO; Civitas Turf Defense™.) and an organosilicone, nonionic soil surfactant (Silwet L-77®) in laboratory and greenhouse bioassays. Civitas and Silwet killed > 75% of ABW adults in multiple assays. The level of control was positively affected by increased rate, spray application volume, and soil moisture levels. Dissections of weevils treated with Civitas revealed material entering the insect's hemocoel after 15−30 min, though most mortality occurred between the 3 and 24 h observation periods. Reducing rates while increasing carrier volume or soil moisture levels through irrigation applied prior to or after application also provided excellent control of adults in the same observation periods. Silwet provided comparable, yet less consistent levels of control in the laboratory studies but was excluded from further tests after treated plants demonstrated phytotoxicity in greenhouse studies. Neither Silwet nor Civitas efficacy was affected by pyrethroid resistance levels in the ABW populations tested.

Entomopathogenic Nematodes for the Management of Plum Curculio in Highbush Blueberry.

Conotrachelus nenuphar Herbst (Coleoptera: Curculionidae) is a key pest of stone and pome fruits in the United States. Application of certain entomopathogenic nematode (EPN) species has shown efficacy in some crops when targeting the larval stage of C. nenuphar in soil. To date, however, no EPNs have been tested for the control of this pest in highbush blueberries. In 2020, laboratory and field studies were conducted to: (1) determine the persistence of Steinernema riobrave, S. carpocapsae, S. feltiae, and Heterorhabditis bacteriophora in acidic blueberry soil; (2) compare the virulence of these EPNs to C. nenuphar larvae and pupae; and (3) compare the efficacy of these EPN species to control this pest in blueberry fields. The greatest persistence in blueberry soil was exhibited by S. riobrave followed by S. carpocapsae. Superior virulence was observed in S. riobrave against C. nenuphar larvae and pupae. Promising levels of virulence were also observed in S. carpocapsae and S. feltiae against the larvae, but S. scarabaei had low virulence. In the field, S. riobrave provided significantly higher levels of C. nenuphar suppression (90%) than the other EPNs. The field efficacy of S. riobrave against C. nenuphar at low and high rates was confirmed in 2021. Steinernema riobrave has the potential to become an important component in the management of C. nenuphar in highbush blueberry.

Diagnostic Dose Assays for the Detection and Monitoring of Resistance in Adults From Listronotus maculicollis (Coleoptera: Curculionidae) Populations.

The annual bluegrass weevil, Listronotus maculicollis (Kirby) (Coleoptera: Curculionidae), is the most difficult to control insect pest on golf courses in eastern North America. Insecticide resistance, particularly to pyrethroids, is a serious and expanding issue in its management. Optimal diagnostic tools for resistance detection are crucial for efficient resistance monitoring and mitigation. Developed vial and Petri dish assays clearly separated different resistance levels among weevil populations. With the pyrethroid bifenthrin, susceptible, moderately resistant (resistance ratios, RR50s 12.2-95.7), and highly resistant (RR50s 258.2-1760.9) populations were distinguished. With the organophosphate chlorpyrifos, susceptible, tolerant (RR50s 2.4-6.7), and resistant (RR50s 8.8-120.7) populations were distinguished. In validation assays, several bifenthrin and chlorpyrifos concentrations were needed to separate resistance levels in Petri dish (bifenthrin: 112.2 and 336.3 or 3,362.5 mg AI/m2; chlorpyrifos: 3.4 and 33.6 mg AI/m2) and vial (bifenthrin: 112.1 or 1,120.8 mg AI/m2; chlorpyrifos: 2.2 and 11.2 mg AI/m2) assays. The Petri dish assay with formulated bifenthrin and chlorpyrifos was the best option for L. maculicollis resistance detection and monitoring. It demonstrated sufficient discriminating power, accurately reflected resistance levels, and was easier to conduct. A single diagnostic concentration sufficed to separate susceptible and resistant populations. To determine different resistance or tolerance levels, two to three concentrations were necessary.

Role of Plant Volatiles in Host Plant Recognition by Listronotus maculicollis (Coleoptera: Curculionidae).

The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby, is an economically important pest of short cut turfgrass. Annual bluegrass, Poa annua L., is the most preferred and suitable host for ABW oviposition, larval survival and development. We investigated the involvement of grass volatiles in ABW host plant preference under laboratory and field conditions. First, ovipositional and feeding preferences of ABW adults were studied in a sensory deprivation experiment. Clear evidence of involvement of olfaction in host recognition by ABW was demonstrated. Poa annua was preferred for oviposition over three bentgrasses, Agrostis spp., but weevils with blocked antennae did not exhibit significant preferences. ABW behavioral responses to volatiles emitted by Agrostis spp. and P. annua were examined in Y-tube olfactometer assays. Poa annua was attractive to ABW females and preferred to Agrostis spp. cultivars in Y-tube assays. Headspace volatiles emitted by P. annua and four cultivars of Agrostis stolonifera L. and two each of A. capillaris L. and A. canina L. were extracted, identified and compared. No P. annua specific volatiles were found, but Agrostis spp. tended to have larger quantities of terpenoids than P. annua. (Z)-3-hexenyl acetate, phenyl ethyl alcohol and their combination were the most attractive compounds to ABW females in laboratory Y-tube assays. The combination of these compounds as a trap bait in field experiments attracted adults during the spring migration, but was ineffective once the adults were on the short-mown turfgrass. Hence, their usefulness for monitoring weevil populations needs further investigation.

Pyrethroid-Resistance Level Affects Performance of Larvicides and Adulticides From Different Insecticide Classes in Populations of Listronotus maculicollis (Coleoptera: Curculionidae).

The annual bluegrass weevil, Listronotus maculicollis Kirby (Coleoptera: Curculionidae), is a major pest of golf course turf in eastern North America with widespread insecticide resistance. This study examined the effect of pyrethroid-resistance level on the efficacy of adulticides and larvicides from different insecticide classes commonly used for L. maculicollis management through greenhouse and field studies. The tested populations had previously been determined to be susceptible, moderately resistant, resistant, and highly resistant to the pyrethroid bifenthrin. Targeting adults, efficacy was significantly reduced for bifenthrin against the highly resistant population and for the spinosyn spinosad and the oxadizine indoxacarb against the resistant and highly resistant populations. Efficacy of the organophosphate chlorpyrifos was not significantly reduced. No adulticide provided significant control of resistant and highly resistant populations. Targeting larvae, the efficacy of spinosad and the anthranilic diamide cyantraniliprole was marginally reduced against the highly resistant population. Significant reductions in efficacy and no significant control were observed for indoxacarb against the highly resistant population and for the neonicotinoid clothianidin, the anthranilic diamide chlorantraniliprole, and the organophosphate trichlorfon against the resistant and highly resistant populations. Our findings lay the groundwork for management recommendations for populations with different resistance levels. Generally, synthetic insecticide applications should be minimized with greater use of larvicides supplemented with nonchemical control alternatives. The remaining effective larvicides should be rotated using cyantraniliprole, spinosad, and indoxacarb against resistant and cyantraniliprole and spinosad against highly resistant populations.

Cross-resistance Patterns to Insecticides of Several Chemical Classes Among Listronotus maculicollis (Coleoptera: Curculionidae) Populations With Different Levels of Resistance to Pyrethroids.

The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby (Coleoptera: Curculionidae), is the most damaging golf course insect pest in eastern North America. Heavy reliance on synthetic insecticides against this pest has led to widespread problems in controlling ABW with pyrethroid resistance already reported from populations in southern New England. This study evaluated the degree and scope of ABW resistance, determined existing cross-resistance patterns, and confirmed laboratory findings under greenhouse conditions. The susceptibility of 10 ABW populations to insecticides of different chemical classes was assessed in topical, feeding, and greenhouse assays. The level of susceptibility to pyrethroids varied significantly among populations (LD50s ranging 2.4-819.1 ng per insect for bifenthrin and 1.1-362.7 ng for λ-cyhalothrin in the topical assay). Three populations were relatively susceptible to pyrethroids, and seven populations had moderate to high resistance levels (RR50 for bifenthrin ranging 30.5-343.1). The toxicity of chlorpyrifos (RR50s ranging 3.3-15.3), spinosad (RR50s 2.4-7.7), clothianidin (RR50s 4.2-9.7), and indoxacarb (RR50s 2.8-9.7) was decreased for the pyrethroid-resistant populations. Toxicity data for bifenthrin and chlorpyrifos obtained under more realistic greenhouse conditions confirmed laboratory observations, indicating that the topical assay is an accurate method of detection and measurement of resistance level. The current study expanded the previously known geographic range of ABW pyrethroid resistance to include the New York metropolitan area, New Jersey, and eastern Pennsylvania and provided clear evidence of cross-resistance not only within the pyrethroid class but also to several other chemical classes.

New Insight into the Management of the Tomato Leaf Miner, Tuta absoluta (Lepidoptera: Gelechiidae) with Entomopathogenic Nematodes.

The tomato leaf miner, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is a serious threat to tomato production in the world. Due to serious issues with insecticide resistance, there is a dire need for alternative control methods. Entomopathogenic nematodes (EPN) have potential for the biological control of T. absoluta. In the laboratory, we examined the effect of temperature, soil type, and exposure time on the efficacy of the EPN species Steinernema carpocapsae (Nematoda: Steinernematidae) and Heterorhabditis bacteriophora (Nematoda: Heterorhabditidae) against last-instar T. absoluta larvae. Both species caused high mortality in loamy sand (89%) and coco peat (93%) but not in sandy loam (17%). H. bacteriophora caused 92-96% mortality at 19, 25, and 31°C; S. carpocapsae caused 89-91% mortality at 25 and 31°C but only 76% at 19°C. Both species caused similar mortality levels after 65-min exposure; thereafter, mortality increased only with S. carpocapsae reaching high levels even at a low concentration. Both species infected larvae within leaf galleries. When applied to whole large tomato plants in the greenhouse, both species provided similar control levels (48-51%) at high pest densities. Both species could be incorporated as an effective alternative to synthetic insecticides into T. absoluta management programs in greenhouse tomato production.

Chilling effect on termination of reproductive diapause in Listronotus maculicollis (Coleoptera: Curculionidae).

Diapausing adults of the annual bluegrass weevil, Listronotus maculicollis, were collected from their hibernating sites at different times in autumn and winter, and subjected to different conditions to observe diapause termination by dissecting and measuring the reproductive organs. When diapausing weevils were maintained under laboratory cold conditions (10 h light at 6 °C:14 h dark at 4 °C) from early December to late March, the sizes of reproductive organs of both sexes increased or fluctuated slightly, and very few females had developing oocytes, suggesting that most adults did not resume development during the chilling period. When diapausing weevils (chilled for 40-83 days) were transferred to warm conditions (LD 14:10 and 21 °C) for different lengths of time, reproductive organ sizes in both sexes increased as chilling period prolonged, implying that chilling played an important role in diapause termination. Under field conditions, an apparent peak of reproductive development was observed on January 07 when 80% of males and 53% of females had resumed growth of reproductive organs. Diapausing weevils collected in September without chilling did not develop successfully despite exposure to warm conditions. In contrast, 87% of males and 93% of females collected from the field on January 21 had initiated reproductive development after 5 days of exposure to warm conditions, indicating the necessity of chilling for diapause termination. Male and female reproductive organ sizes increased faster and to a greater final size the longer the preceding chilling period was. The prolonged chilling period in the field resulted in more synchronized and advanced development in L. maculicollis when exposed to warm conditions.

Synergistic Combinations of a Pyrethroid Insecticide and an Emulsifiable Oil Formulation of Beauveria bassiana to Overcome Insecticide Resistance in Listronotus maculicollis (Coleoptera: Curculionidae).

The annual bluegrass weevil, Listronotus maculicollis (Kirby), is a major pest of golf course turf in eastern North America and has become particularly problematic owing to widespread development of insecticide resistance. As an alternative option to manage resistant adult L. maculicollis, we explored combinations of the pyrethroid insecticide bifenthrin with an emulsifiable oil formulation of the entomopathogenic fungus Beauveria bassiana strain GHA (Bb ES). Combinations synergistically enhanced mortality in both insecticide-susceptible and insecticide-resistant L. maculicollis adults in the laboratory when bifenthrin was used at LC50s for each population. To determine the component behind the synergism, technical spores of B. bassiana GHA and the emulsifiable oil carrier in the fungal formulation were tested separately or in combination with bifenthrin. In both separate and combined applications, the emulsifiable oil carrier was responsible for high mortality within 3 d after treatment and interacted synergistically with bifenthrin, whereas fungus-induced mortality started later. Strong synergism was also observed in three field experiments with a relatively resistant L. maculicollis population. Combinations of Bb ES and bifenthrin hold promise as an effective L. maculicollis management tool, particularly of pyrethroid-resistant populations.

Differential Response of a Local Population of Entomopathogenic Nematodes to Non-Native Herbivore Induced Plant Volatiles (HIPV) in the Laboratory and Field.

Recent work has shown the potential for enhanced efficacy of entomopathogenic nematodes (EPN) through their attraction to herbivore induced plant volatiles. However, there has been little investigation into the utilization of these attractants in systems other than in those in which they were identified. We compared (E)-ß-caryophyllene and pregeijerene in the highbush blueberry (Vaccinium corymbosum) agroecosystem in their ability to enhance the attraction of EPN to and efficacy against the system's herbivore, oriental beetle (Anomala orientalis). The relative attractiveness of (E)-ß-caryophyllene and pregeijerene to a local isolate of the EPN species Steinernema glaseri was tested in a six-arm olfactometer in the laboratory to gather baseline values of attraction to the chemicals alone in sand substrate before field tests. A similar arrangement was used in a V. corymbosum field by placing six cages with assigned treatments and insect larvae with and without compound into the soil around the base of 10 plants. The cages were removed after 72 h, and insect baits were retrieved and assessed for EPN infection. The lab results indicate that in sand alone (E)-ß-caryophyllene is significantly more attractive than pregeijerene to the local S. glaseri isolate Conversely, there was no difference in attractiveness in the field study, but rather, native S. glaseri were more attracted to cages with G. mellonella larvae, no larvae, and cages with the blank control and G. mellonella larvae.

Horizontal gene transfer of a bacterial insect toxin gene into the Epichloë fungal symbionts of grasses.

Horizontal gene transfer is recognized as an important factor in genome evolution, particularly when the newly acquired gene confers a new capability to the recipient species. We identified a gene similar to the makes caterpillars floppy (mcf1 and mcf2) insect toxin genes in Photorhabdus, bacterial symbionts of nematodes, in the genomes of the Epichloë fungi, which are intercellular symbionts of grasses. Infection by Epichloë spp. often confers insect resistance to the grass hosts, largely due to the production of fungal alkaloids. A mcf-like gene is present in all of the Epichloë genome sequences currently available but in no other fungal genomes. This suggests the Epichloë genes were derived from a single lineage-specific HGT event. Molecular dating was used to estimate the time of the HGT event at between 7.2 and 58.8 million years ago. The mcf-like coding sequence from Epichloë typhina subsp. poae was cloned and expressed in Escherichia coli. E. coli cells expressing the Mcf protein were toxic to black cutworms (Agrotis ipsilon), whereas E. coli cells containing the vector only were non-toxic. These results suggest that the Epichloë mcf-like genes may be a component, in addition to the fungal alkaloids, of the insect resistance observed in Epichloë-infected grasses.

Storage temperature and duration affect Steinernema scarabaei dispersal and attraction, virulence, and infectivity to a white grub host.

The entomopathogenic nematode Steinernema scarabaei has exceptional potential for the control of many white grub species. Initial studies suggested this species to have a widely ranging foraging strategy based on its attraction to hosts in soil columns, however, with a generally low but very variable dispersal rate even in the presence of hosts. The objective of this study was to develop a better understanding of the dispersal behavior of S. scarabaei IJs and the influence of storage conditions on its dispersal and infectivity. We found that storage temperature and duration had a strong effect on S. scarabaei IJ dispersal, virulence, and infectivity. But even under conditions conducive to movement only a small proportion of IJs moved towards a host. IJ dispersal declined with storage time by a factor of around 100 between 1 week and 12 weeks of storage whether the IJs were stored at room temperature or 8°C; however, the decline was more than twice as fast after storage at 8°C. Host attraction also diminished with storage duration. IJ virulence and infectivity declined with storage time for IJs stored at room temperature. In contrast, for IJs stored at 8°C virulence remained high and infectivity increased over time. The decrease in dispersal and infectivity when stored at room temperature may reflect an adaptation to conserve energy in the absence of hosts since S. scarabaei IJs have to persist through extended periods in summer during which infections are unlikely to occur. The even faster decrease in dispersal rate when stored at 8°C may suggest a cold-induced dormancy that may serve as an overwintering strategy. The parallel increase in infectivity, however, seems to contradict such a strategy. Future studies should examine whether and how S. scarabaei IJs 'inactivated' by the absence of hosts can be 'reactivated' and whether this behavior could be used to improve the efficacy of nematode applications against insect pests.

Subterranean, herbivore-induced plant volatile increases biological control activity of multiple beneficial nematode species in distinct habitats.

While the role of herbivore-induced volatiles in plant-herbivore-natural enemy interactions is well documented aboveground, new evidence suggests that belowground volatile emissions can protect plants by attracting entomopathogenic nematodes (EPNs). However, due to methodological limitations, no study has previously detected belowground herbivore-induced volatiles in the field or quantified their impact on attraction of diverse EPN species. Here we show how a belowground herbivore-induced volatile can enhance mortality of agriculturally significant root pests. First, in real time, we identified pregeijerene (1,5-dimethylcyclodeca-1,5,7-triene) from citrus roots 9-12 hours after initiation of larval Diaprepes abbreviatus feeding. This compound was also detected in the root zone of mature citrus trees in the field. Application of collected volatiles from weevil-damaged citrus roots attracted native EPNs and increased mortality of beetle larvae (D. abbreviatus) compared to controls in a citrus orchard. In addition, field applications of isolated pregeijerene caused similar results. Quantitative real-time PCR revealed that pregeijerene increased pest mortality by attracting four species of naturally occurring EPNs in the field. Finally, we tested the generality of this root-zone signal by application of pregeijerene in blueberry fields; mortality of larvae (Galleria mellonella and Anomala orientalis) again increased by attracting naturally occurring populations of an EPN. Thus, this specific belowground signal attracts natural enemies of widespread root pests in distinct agricultural systems and may have broad potential in biological control of root pests.

Entomopathogenic nematodes for the management of Agrotis ipsilon: effect of instar, nematode species and nematode production method.

BACKGROUND: Previous laboratory studies have indicated the potential of some entomopathogenic nematode (EPN) species for the control of larvae of the black cutworm (BCW). To determine the most promising EPN species and the most susceptible BCW stages, a more in-depth evaluation of seven EPN species against different BCW instars was carried out, the efficacies of in vitro- and in vivo-produced EPNs were compared and the suitability of BCW instars for EPN reproduction was examined. RESULTS: Heterorhabditis megidis was the most virulent species, irrespective of larval stage in small arenas, followed most often by H. bacteriophora. In pots with grass, Steinernema carpocapsae tended to be the most virulent species, followed by H. bacteriophora, H. megidis and S. riobrave. Fourth and/or fifth instars were the most susceptible stages to most EPN species, and pupae the least susceptible. Furthermore, H. bacteriophora, H. megidis and S. carpocapsae successfully reproduced in fifth and sixth instars and pupae. In vivo-produced H. megidis and S. carpocapsae controlled fifth instars better than the corresponding in vitro-produced products; production method did not affect H. bacteriophora and S. riobrave efficacy. CONCLUSIONS: Several in vitro-produced commercial EPN strains were highly virulent to BCW and warrant further testing under field conditions, along with some in vivo-produced strains.

Development of binomial sequential sampling plans for forecasting Listronotus maculicollis (Coleoptera: Curculionidae) larvae based on the relationship to adult counts and turfgrass damage.

Binomial sequential sampling plans were developed to forecast weevil Listronotus maculicollis Kirby (Coleoptera: Curculionidae), larval damage to golf course turfgrass and aid in the development of integrated pest management programs for the weevil. Populations of emerging overwintered adults were sampled over a 2-yr period to determine the relationship between adult counts, larval density, and turfgrass damage. Larval density and composition of preferred host plants (Poa annua L.) significantly affected the expression of turfgrass damage. Multiple regression indicates that damage may occur in moderately mixed P. annua stands with as few as 10 larvae per 0.09 m2. However, > 150 larvae were required before damage became apparent in pure Agrostis stolonifera L. plots. Adult counts during peaks in emergence as well as cumulative counts across the emergence period were significantly correlated to future densities of larvae. Eight binomial sequential sampling plans based on two tally thresholds for classifying infestation (T = 1 and two adults) and four adult density thresholds (0.5, 0.85, 1.15, and 1.35 per 3.34 m2) were developed to forecast the likelihood of turfgrass damage by using adult counts during peak emergence. Resampling for validation of sample plans software was used to validate sampling plans with field-collected data sets. All sampling plans were found to deliver accurate classifications (correct decisions were made between 84.4 and 96.8%) in a practical timeframe (average sampling cost < 22.7 min).

Sex pheromone of the scarab beetle Phyllophaga (Phytalus) georgiana (horn).

The sex pheromone of Phyllophaga (Phytalus) georgiana was characterized as valine methyl ester, tentatively the L-enantiomer. This is the first sex pheromone identified from the Phyllophaga subgenus Phytalus. The pheromone was extracted from female glands, the active component isolated by coupled gas chromatography-electroantennogram detection analysis, characterized by mass spectrometry, and shown to be active in field tests. The seasonal flight pattern was determined for P. georgiana as well as for three other species, P. anxia (both northern and southern genitalic forms), P. gracilis, and P. postrema. The latter three species were captured in traps baited with L-isoleucine methyl ester.

Pellet formulations of sex pheromone components for mating disruption of oriental beetle (Coleoptera: Scarabaeidae) in turfgrass.

Wax-based pelleted sex pheromone formulations were tested for efficacy in managing oriental beetle, Anomala orientalis Waterhouse, in turfgrass. Four pellet formulations were field tested at 25 g pheromone/ha during 2006 and 2007. Application patterns included pellets specifically placed in a uniform grid pattern and pellets scattered by a broadcast spreader. Treatment efficacy was measured by monitoring male A. orientalis captures in pheromone-baited traps, determining mating success of confined virgin females, and estimating subsequent densities of A. orientalis larvae in soil/sod samples. All formulation and application pattern combinations effectively suppressed mating for 2 wk after application as measured by reduction in male trap captures and mating success of confined virgins. Two formulations gradually lost efficacy beginning 2 wk after application and the two others after 3-4 wk. Application pattern had no effect on trap captures. The most effective formulation reduced trap captures by >/=90% for 36 d, reduced mating success of confined females by 86-100% until 24 d after treatment, and reduced A. orientalis larval populations by 69%. In a previous study, sprayable formulations resulted in nuisance contamination of shoes that attracted male beetles outside of pheromone-treated areas. In contrast, shoes walked through pellet-treated areas did not attract male beetles. Pellet formulations have great potential for A. orientalis mating disruption, but more research is needed to optimize performance.

Attraction of four entomopathogenic nematodes to four white grub species.

To better understand the differences in the efficacy of entomopathogenic nematode species against white grub species, we are studying the various steps of the infection process of entomopathogenic nematodes into different white grub species using nematode species/strains with particular promise as white grub control agents. In this study we compared the attraction of the entomopathogenic nematodes Steinernema scarabaei (AMK001 strain), Steinernema glaseri (NC1 strain), Heterorhabditis zealandica (X1 strain), and Heterorhabditis bacteriophora (GPS11 strain) to third-instars of the scarabs Popillia japonica, Anomala orientalis, Cyclocephala borealis, and Rhizotrogus majalis, and late-instar greater wax moth, Galleria mellonella, larvae. Individual larvae were confined at the bottom of 5.5 cm vertical sand columns, nematodes added to the sand surface after 24 h, and nematodes extracted after another 24 h. Nematode attraction to hosts was strongly affected by nematode species but the effect of insect species varied with nematode species. S. glaseri had a high innate dispersal rate (i.e., in absence of insects) and was strongly attracted to insects without significant differences among insect species. S. scarabaei had a very low innate dispersal rate so that even a strong relative response to insects resulted in low absolute dispersal rates toward insects. S. scarabaei tended to be most attracted to G. mellonella and least attracted to C. borealis. H. zealandica had a high innate dispersal rate but only responded weakly to insects without significant differences among species. H. bacteriophora had limited innate dispersal and only weakly responded to insects with G. mellonella tending to be the most attractive and C. borealis the least attractive insect. It has to be noted that we cannot exclude that the use of different rearing hosts (A. orientalis and P. japonica larvae for S. scarabaei, G. mellonella larvae for the other nematodes) might have had an impact on the nematodes dispersal and relative attraction behavior. This study indicates that host attractiveness and nematode dispersal rates may contribute but do not play a major role in the variability in white grub susceptibility and/or nematode virulence.