Effect of insect cadaver desiccation and soil water potential during rehydration on entomopathogenic nematode (Rhabditida: Steinernematidae and Heterorhabditidae) production and virulence.

We examined the influence of insect cadaver desiccation on the virulence and production of entomopathogenic nematodes (EPNs), common natural enemies of many soil-dwelling insects. EPNs are often used in biological control, and we investigated the feasibility of applying EPNs within desiccated insect cadavers. Desiccation studies were conducted using the factitious host, Galleria mellonella (Lepidoptera: Pyralidae, wax moth larvae) and three EPN species (Heterorhabditis bacteriophora 'HB1', Steinernema carpocapsae 'All', and Steinernema riobrave). Weights of individual insect cadavers were tracked daily during the desiccation process, and cohorts were placed into emergence traps when average mass losses reached 50%, 60%, and 70% levels. We tracked the proportion of insect cadavers producing infective juveniles (IJs), the number and virulence of IJs produced from desiccated insect cadavers, and the influence of soil water potentials on IJ production of desiccated insect cadavers. We observed apparent differences in the desiccation rate of the insect cadavers among the three species, as well as apparent differences among the three species in both the proportion of insect cadavers producing IJs and IJ production per insect cadaver. Exposure of desiccated insect cadavers to water potentials greater than -2.75 kPa stimulated IJ emergence. Among the nematode species examined, H. bacteriophora exhibited lower proportions of desiccated insect cadavers producing IJs than the other two species. Desiccation significantly reduced the number of IJs produced from insect cadavers. At the 60% mass loss level, however, desiccated insect cadavers from each of the three species successfully produced IJs when exposed to moist sand, suggesting that insect cadaver desiccation may be a useful approach for biological control of soil insect pests.

Evolution of host search strategies in entomopathogenic nematodes.

There is interspecific variation in infective juvenile behavior within the entomopathogenic nematode genus Steinernema. This variation is consistent with use of different foraging strategies along a continuum between ambush and cruise foraging. To address questions about the evolution of foraging strategy, behavioral and morphological characters were mapped onto a phylogeny of Steinernema. Three species, all in the same clade, were classified as ambushers based on standing bout duration and host-finding ability. One clade of six species were all cruisers based on both host-finding and lack of standing behavior. All species in the ambusher clade had a high rate of jumping, all species in the cruiser clade had no jumping, and most intermediate foragers exhibited some level of jumping. Response to volatile and contact host cues was variable, even within a foraging strategy. Infective juveniles in the ambusher clade were all in the smallest size category, species in the cruiser clade were in the largest size categories, and intermediate foragers tended to be more intermediate in size. We hypothesize that the ancestral Steinernema species was an intermediate forager and that ambush and cruise foraging both evolved at least once in the genus.

Developmental temperature effects on five geographic isolates of the entomopathogenic nematode Steinernema feltiae (Nematoda: Steinernematidae).

The development of five geographic isolates of Steinernema feltiae at 5, 8, 10, 15, 20, 25, and 28 degrees C in wax moth, Galleria mellonella, larvae was examined. The isolates were from Mediterranean (Sinop from Turkey, SN from France, and Monterey from California), subtropical (Rafaela from Argentina), and tropical (MG-14 from Hawaii) regions. All isolates caused 100% mortality of wax moth larvae and developed and produced progeny between 8 and 25 degrees C. At 28 degrees C, mortality was 100%, but no progeny was observed. The highest infective juvenile production was observed at 15 degrees C for all isolates. In general, the tropical isolate, MG-14, had the lowest production of infective juveniles. The time of emergence of the infective juveniles from the host cadaver showed some differences among isolates, with the Sinop isolate having the earliest emergence time from cadavers at 15 degrees C (10 days) and 20 degrees C (8 days). At 25 degrees C, the infective juveniles of the Sinop, SN, and Rafaela isolates emerged from the cadavers from 5 to 7 days. Time of host death by all isolates showed no differences at 8, 10, 15, 20, and 28 degrees C. At 25 degrees C for all isolates (except the MG-14), shorter times to host death were observed. Host death occurred at 12 days at 8 degrees C, 9 to 11 days at 10 degrees C, 4 to 5 days at 15 degrees C, 3 days at 20 degrees C, and 2 days at 25 and 28 degrees C. For penetration efficiency, the Sinop, SN, and Rafaela isolates penetrated their hosts at 5, 8, and 10 degrees C. Penetration of the infective juveniles was consistently high for all isolates at 15, 20, 25, and 28 degrees C, but it was significantly lower for the MG-14 isolate at 15, 25, and 28 degrees C. No progeny production occurred at 28 degrees C, but nematode penetration did occur with the MG-14 isolate having significantly lower penetration than the other isolates. When nematodes were produced at 8, 15, and 23 degrees C in wax moth larvae, all isolates had infective juveniles with longer body lengths at 8 degrees C followed by 15 and 23 degrees C. To further verify body length at the different temperatures, beet armyworm, Spodoptera exigua, larvae and dog-food agar medium were used, respectively, for in vivo and in vitro culture of the Sinop isolate. Infective juvenile body length showed the same trends, with the longest being at 8 degrees C and decreasing in length from 15 to 23 degrees C. The data suggest that quality of food for the nematode and temperature (that is, developmental time) influence the body length of the infective juvenile.

Biological control agents for white grubs (Coleoptera: Scarabaeidae) in anticipation of the establishment of the Japanese beetle in California.

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.

Are there temporarily non-infectious dauer stages in entomopathogenic nematode populations: a test of the phased infectivity hypothesis.

Many studies of entomopathogenic nematodes (Heterorhabditidae and Steinernematidae) have reported that only a small proportion (typically < 40%) of infective stages (dauers), even under apparently ideal conditions, actually infect a host. The 'phased infectivity hypothesis' is most frequently invoked to explain this pattern of low infection with entomopathogenic nematodes. It proposes that at a given point in time not all individuals are infectious i.e. infectiousness is delayed in some individuals. We tested experimentally several predictions based on this hypothesis. Specifically, if phased infectivity occurs, we should be able to expose dauers to increasing numbers of potential hosts until dauers no longer infect and still be able to recover viable dauers. These recovered dauers which did not infect should be infectious at some later point in time. However, our results do not support the phased infectivity hypothesis for 3 species of Steinernema: most dauers could be recovered in one sampling round when provided with sufficient suitable hosts. In contrast, Heterorhabditis bacteriophora frequently did not infect all available hosts, and infectious dauers were recovered in subsequent sampling rounds. This result is more consistent with the phased infectivity hypotheses, but further research is needed before we can be more confident in the hypothesis. For all species tested, the number of available hosts influenced population levels of nematode infectivity. This suggests that the infection status of hosts can influence whether a dauer infects. Our results indicate that phased infectivity is not a common phenomenon in entomopathogenic nematode dauers, despite the widespread acceptance of this hypothesis.

Suppression of diamondback moth (Lepidoptera: Plutellidae) with an entomopathogenic nematode (Rhabditida: Steinernematidae) and Bacillus thuringiensis Berliner.

We tested the efficacy of the All strain of Steinernema carpocapsae (Weiser) against larvae of the diamondback moth, Plutella xylostella (L.). In laboratory bioassays we found that (1) commercially formulated nematodes produced in vitro were as effective as nematodes produced in vivo, (2) resistance of P. xylostella to Bacillus thuringiensis Berliner subsp. kurstaki did not confer cross-resistance to nematodes, (3) mortality caused by nematodes was higher for early than late 3rd-instar P. xylostella larvae, and (4) no interaction occurred when B. thuringiensis and nematodes were combined against a susceptible strain of P. xylostella, but an antagonistic interaction occurred between the 2 pathogens against a strain of P. xylostella resistant to B. thuringiensis. In field trials conducted on 2 watercress [Rorippa Nasturtium-aquaticum (L.) Hayek] farms in Hawaii, nematodes provided 41% control, B. thuringiensis subsp. aizawai gave 44% control, and the combined treatment (B. thuringiensis plus nematodes both at half rate) resulted in 58% control. Using nemodes to control diamondback moth can theoretically reduce resistance development in diamondback moth populations to B. thuringiensis products, but repeated applications of nematodes will probably be ineffective in attaining control (suggested in simulation model). The results of this study demonstrate that nematodes may be a useful component of integrated pest management programs if efficacy can be increased, especially for populations of P. xylostella that are resistant to B. thuringiensis.

Efficacy of a dehydrated steinernematid nematode against black cutworm (Lepidoptera: Noctuidae) and diamondback moth (Lepidoptera: Plutellidae).

We compared the ability of in vitro-produced, commercially formulated with in vivo-produced, nonformulated Steinernema carpocapsae (Weiser) Poinar. All strain to infect and kill larvae of black cutworm, Agrotis ipsilon (Hufnagel), and diamondback moth, Plutella xylostella (L.). In vitro-produced nematodes formulated in wettable dispersible granules, which were stored dry, were rehydrated in water for 0-72 h before application. Against black cutworms, the efficacy of nematodes was (from most to least effective): in vivo > in vitro rehydrated for 72 h > in vitro rehydrated for 48 h > in vitro rehydrated for 24 h > dehydrated (0 h). Nematodes rehydrated for 72 h in water or moist soil were equally effective against black cutworm larvae, and both were significantly more effective than nematodes without rehydration. These results indicated that nematodes in the wettable dispersible granule formulation required time to rehydrate in the soil before infecting black cutworm larvae. Nematode treatments described above were applied to radish plants held at 100 or 75% RH and tested against diamondback moth larvae. At 100% RH, nematode efficacy was (from most to least effective): in vitro rehydrated for 72 h > in vivo > in vitro rehydrated 48 h > in vitro rehydrated 24 h > dehydrated (0 h). The efficacy of all treatments was lower at 75% than at 100% RH, and the ranking of in vivo and in vitro nematodes rehydrated for 72 h was reversed. The nematodes in the wettable dispersible granule formulation were effective for foliar treatments when humidity was high and nematodes were rehydrated for at least 48 h before application. The data show that nematode infectivity was reduced unless nematodes were rehydrated.

Instar susceptibility of the monarch butterfly (Danaus plexippus) to the neogregarine parasite, Ophryocystis elektroscirrha.

The susceptibility of the monarch butterfly (Danaus plexippus) larvae to the neogregarine parasite, Ophryocystis elektroscirrha, was tested in the laboratory. Spore loads recovered from infected monarch butterflies were directly related to the inoculum level, larval stage of the host, and spore age. There was a linear relationship between spores ingested by first instar larvae and spore concentration. Larvae feeding on leaves treated with 0, 50, 500, 5000, or 50,000 spores averaged 0, 0, 193, 457, or 1,255 spores, respectively, on the abdomens of the adult butterflies. When first, third, and fifth instar larvae were given 14.5 spores/mg of body weight, there was no significant difference in the spore load of the adults resulting from the first and third instars. However, there were significant differences in the spore load from adults resulting from the first and third instars versus the fifth instar. In addition, 1-year-old spores were not as infectious as fresh spores. Our findings indicate that under field conditions, the first instar is most likely to become infected because one spore appears sufficient to produce a detectable spore load in the adult. Older instars are less susceptible and have fewer opportunities to encounter sufficient viable spores for infection to occur. Thus, vertical transmission appears to be the primary mode of parasite maintenance in natural populations of monarch butterflies.

Effect of nematode-trapping fungi on an entomopathogenic nematode originating from the same field site in California.

We determined whether nematode-trapping fungi may influence the dynamics of a coastal shrub community. The food chain interactions in the shrub community involve the dominant plant species, its major insect herbivore, and an entomopathogenic nematode, Heterorhabditis hepialus. Of the 12 nematode-trapping fungi previously isolated from soils at the study site, 5 were selected for this study. Arthrobotrys oligospora, Geniculifera paucispora, Monacrosporium eudermatum, and Monacrosporium cionopagum efficiently trapped and colonized H. hepialus on agar; in contrast Nematoctonus concurrens trapped but did not infect or colonize the nematode on agar. To determine whether these fungi can suppress H. hepialus in soil, we added the fungi in the form of fungal-colonized nematodes to pasteurized (2 hr at 62 degrees C) and raw (nontreated) soil from the study site. Suppression was measured by comparing nematode invasion into a wax moth larva in fungus-treated and untreated soil in vials at 20 degrees C. Fungal population density in soil was estimated using dilution plating and most probable number procedures. All fungi suppressed H. hepialus if the wax moth larvae were added 4 days after the nematodes. Suppression ranged between 37 and 54% and did not differ among fungi. Suppression was usually greater in raw than in pasteurized soil. Raw soil contained a constant background of nematode-trapping fungi, and A. oligospora was the most common among these; no background was detected in pasteurized soil. The presence of background fungi in raw soil may explain the higher suppression in raw than in pasteurized soil. Fungal propagule densities in our laboratory experiments were similar to those observed in the field, suggesting that nematode-trapping fungi may influence the dynamics of the plant, insect herbivore, and entomopathogenic nematode in the coastal ecosystem.

A multivariate analysis of morphometric characters of Heterorhabditis species (Nemata: Heterorhabditidae) and the role of morphometrics in the taxonomy of species of the genus.

A multivariate analysis on the morphometrics of 7 Heterorhabditis species was conducted to review the status of the traditional morphometric characterization methods for differentiating species of these nematodes. Results from this study showed consistency in the selection of morphometric characters for discriminating among males and infective juveniles of Heterorhabditis species. For the males, testis reflexion (TREF) and total length (LENGTH) were the variables that contributed most in the discrimination among the different species, and for the infective juveniles, tail length (TAILL) and total length (LENGTH) were the variables that contributed most. Therefore, we consider that these morphometric characters are useful and reliable, and that they should be used for the identification of Heterorhabditis species/isolates.

Larvicidal activity of the symbiotic bacterium Xenorhabdus japonicus from the entomopathogenic nematode Steinernema kushidai against Anomala cuprea (Coleoptera:Scarabaeidae).

The entomopathogenicity of the symbiotic bacterium Xenorhabdus japonicus and the nematode Steinernema kushidai was determined. Phase I and II X. japonicus were cultured on an artificial medium and inoculated into the test insect or established into axenic S. kushidai populations. When 100, 1000, or 10,000 bacterial cells of phase I or II were directly injected into the hemocoels of 3rd instar cupreous chafer, Anomala cuprea, both phases in the late log period killed 100% of the larvae by the 2nd day postinoculation. However, both phases in the stationary period were less pathogenic with cupreous chafer mortality < 20 and 80% at 100 and 1000 bacterial cells/ larva, respectively. In vitro studies showed that axenic S. kushidai provided with phase I or II symbionts grew well and produced equal numbers of progeny on a dog food medium, but nematodes with no symbionts did not grow at all. Pig liver extracts added as a dietary supplement to the dog food medium completely restored growth and progeny production of the nematode with no bacterial cells. Studies were conducted with infective juveniles (IJs) harboring phase I or II or no symbionts that were applied against 3rd instar cupreous chafer larvae in compost or injected directly into their hemocoels. In the compost study, IJs harboring phase I killed 100% within 10 days. IJs with phase II or no symbionts caused low mortality of the cupreous chafer larvae at 10 days (< 20%). In the intrahemocoelic injection study, IJs harboring phase I resulted in 60% larval mortality at five nematodes/larva, and as the number of IJs injected increased, significantly higher larval mortality was obtained.

Competition between two steinernematid nematode species for an insect host at different soil depths.

We studied interactions between 2 entomopathogenic nematode species, Steinernema carpocapsae, an ambusher forager, and Steinernema glaseri, a cruiser forager, when they were provided wax moth larvae as hosts at 0, 2, or 10 cm soil depth. Populations of infective juvenile nematodes in soil were monitored at 30-day intervals over 120 days using wax moth larvae as baits. After application of S. carpocapsae, S. glaseri, or the combination of both species, hosts were added at 30-day intervals. With hosts at 0 cm depth, each nematode species was negatively affected by the presence of the other species at the 30- and 60-day samples. At 90 and 120 days, S. carpocapsae numbers in the combined treatment were as high as in the single species treatment, whereas only few S. glaseri were recovered. With hosts at 2 or 10 cm depth, the presence of S. glaseri had a strong negative effect on S. carpocapsae, but S. glaseri was not affected by the presence of S. carpocapsae. In another experiment, S. carpocapsae dominated over S. glaseri in hosts located at 0 cm depth as measured by penetration efficiency into hosts and progeny production. In contrast, S. glaseri dominated at 2 cm depth. At 2 cm depth, S. carpocapsae penetrated into hosts too slowly to compete successfully with S. glaseri. Steinernema carpocapsae is superior to S. glaseri when competing for a host on the soil surface; however, below the surface S. glaseri is superior to S. carpocapsae.

Entomopathogenic nematodes: natural enemies of root-feeding caterpillars on bush lupine.

A new species of soil-dwelling entomopathogenic nematode Heterorhabditis hepialus killed up to 100% (mean=72%) of root-boring caterpillars of a ghost moth Hepialus californicus in coastal shrub lands. When unchecked, ghost moth caterpillars killed bush lupine, Lupinus arboreus. Here we describe this strange food chain. Although unappreciated by ecologists, entomopathogenic nematodes are widespread and probably one of the most important groups of natural enemies for underground insects. The free-living infective juvenile (IJ) of entomopathogenic nematodes searches for host insects in the soil. A single IJ can kill a host, although several often invade together. After entering the host through a spiracle or other orifice, the IJ regurgitates its symbiotic bacterium, Photorhabdus luminescens, which kills the host within 48 h. The bacteria digest the cadaver and provide food for the exponentially growing nematode population inside. The bacteria produce antibiotics and other noxious substances that protect the host cadaver from other microbes in the soil. When the cadaver is exhausted of resources, IJs break the host integument and can disperse. As many as 420,000 IJs can be produced within a large ghost moth caterpillar. Surface soil of the lupine rhizosphere is the primary habitat of IJs of H. hepialus. Attracted to waste gases emitted by insects, the 0.5-mm-long IJs can move 6 cm/day through moist soil. Prevalences of H. hepialus ranged from as high as 78% of rhizospheres in some lupine stands to almost zero in others, but it was absent from no stand at our study site. Field intensities ranged from 0.003 IJs/cm3 of soil to 7.5 IJs/cm3, and correlated roughly with prevalences among sites. Few ghost moth caterpillars (mean=6.7) succeeded in entering lupine roots where prevalence of H. hepialus was highest, and this stand had lowest mortality (0.02) of mature bush lupine. In the three stands with lowest prevalence (mean = 2%) of this nematode, many caterpillars (mean = 38.5) entered roots, and lupine mortality was high (range = 0.41-1.0). Old aerial photographs indicate that the stands with highest recent nematode prevalence have had little or no mass die-off of lupine over the past 40 years. The photos depict repeated die-offs of lupine during the past four decades in stands with lowest recent prevalence of the nematode. This pattern leads us to entertain the hypothesis that the nematode affects vegetation dynamics indirectly through a trophic cascade. Dispersal of entomopathogenic nematodes is little understood. We found that air drying of soil extirpates H. hepialus and speculate that this nematode is dispersed during the wet season in moist soil bits on the exterior of fossorial insects and mammals. H. hepialus colonized some previously unoccupied lupine rhizospheres during the wet winter-spring season and, obversely, became extinct from some rhizosperes as soil dried in summer. Root-feeding insects have only recently been recognized as a force in communities, and the regulation of these important herbivores is still largely an ecological terra incognita. All evidence indicates that entomopathogenic nematodes are found throughout terrestril ecosystems, and we propose that trophic chains similar to those described in this report should not be uncommon.

Density-dependent effects on Steinernema glaseri (Nematoda: Steinernematidae) within an insect host.

Increasing densities of Steinernema glaseri infective juveniles (IJs) in soil affected penetration efficiency and reproduction of the nematodes in larvae of the greater wax moth Galleria mellonella. The penetration efficiency and the proportion of penetrated IJs developing to adults decreased significantly with increasing numbers of IJs present in the soil and entering the hosts, respectively. The number of progeny produced/host cadaver initially increased, with the highest production being between 20.7 +/- 3.3 and 58.0 +/- 6.6 IJs established/host +/- SE and decreased at higher densities. Above 184.4 +/- 37.7 IJs established/host, no progeny emerged from the cadavers.

Lagenidium giganteum in Culex tarsalis larvae: production of infective propagules.

Zoospore and oospore production of the entomopathogenic fungus, Lagenidium giganteum, was quantified from Culex tarsalis larvae in rice irrigation water. For the first through fourth instars, respectively, the average zoospore production (+/- SE) was 521 +/- 106, 2458 +/- 365, 10,546 +/- 582, and 14,360 +/- 1076, and the average oospore production was 17 +/- 1.8, 33 +/- 3.5, 44 +/- 2.6, and 34 +/- 3.1. Zoosporogenesis varied with the water sample. The ionic composition of the water samples was significantly correlated with zoospore production. The correlations, however, changed as different subsets of data were analyzed, suggesting that other factors also affected zoosporogenesis. The extent to which a cadaver was colonized by other microorganisms had a clear effect on the resulting Lagenidium reproduction. The interval of peak zoosporogenesis occurred between 24 and 36 hr postmortem for the first instar and 12 to 24 hr postmortem for other instars. Measurable production was observed up to 96 hr postmortem from fourth instar larvae.

Nonsusceptibility of the Honey Bee, Apis mellifera (Hymenoptera: Apidae), to Steinernematid and Heterorhabditid Nematodes.

We exposed honey bee workers and brood to four entomopathogenic nematode species under conditions normally encountered in the hive by spraying nematodes onto combs. Mortality of adult bees exposed to any of the nematode species was less than 10%, and there was no evidence of nematode infection when dead adults were dissected. To assess the impact of nematodes on brood, we used smaller-size honey combs placed in the second story (super) of a hive and large brood combs placed in the main section of the hive. Our results were inconsistent between these two experimental designs. The smaller honey combs sprayed with Steinernema carpocapsae contained the largest number of uncapped ceils, those sprayed with Heterorhabditis baeteriophora or S. riobravis contained an intermediate number of uncapped cells, and control combs and those sprayed with S. glaseri contained the fewest nmnber of uncapped cells. Large combs sprayed with S. riobravis contained more uncapped ceils than controls or those sprayed with S. carpocapsae, although the differences were not significant. Our results do not support the hypothesis that high-temperature-tolerant species of nematodes are necessarily more infective to honey bees.

A new species of Heterorhabditis from the Hawaiian Islands.

A new species of nematode of the genus Heterorhabditis (Nemata: Heterorhabditidae) was found during a survey of the soil entomopathogenic nematode fauna of the Hawaiian Islands. Heterorhabditis hawaiiensis sp. n. can be separated from all other species of Heterorhabditis by the length of the infective juvenile and the morphological characters of the spicules, gubernaculum, and bursa. Random amplified polymorphic DNA (RAPD) fragment analysis showed that this species also has a distinct genetic pattern in RAPD bands relative to the other 6 species or isolates of Heterorhabditis that were compared.

Influence of salinity on survival and infectivity of entomopathogenic: nematodes.

Exposure to NaC1, KCI, and CaCl affected the entomopathogenic nematodes Heterorhabditis bacteriophora and Steinernema glaseri differently. Survival, virulence, and penetration efficiency of S. glaseri were not affected by these salts. At high concentrations, however, all three salts inhibited its ability to move through a soil column and locate and infect a susceptible host. Calcium chloride and KCl had no effect on H. bacteriophora survival, penetration efficiency, or movement through a soil column, but moderate concentrations of these salts enhanced H. bacteriophora virulence. NaCl, however, adversely affected each of these parameters at high salinities (>16 dS/m). Salt effects on S. glaseri are attributed solely to interference with nematode host-finding ability, whereas the NaCl effects on H. bacteriophora are attributed to its toxicity and possibly to interference with host-finding behavior.

Impact of a Nematode-parasitic Fungus on the Effectiveness of Entomopathogenic Nematodes.

The impact of the nematode-parasitic fungus Hirsutella rhossiliensis on the effectiveness of Steinernema carpocapsae, S. glaseri, and Heterorhabditis bacteriophora against Galleria mellonella larvae was assessed in the laboratory. The presence of Hirsutella conidia on the third-stage (J3) cuticle of S. carpocapsae and H. bacteriophora interfered with infection of insect larvae. Conidia on the J3 cuticle of S. glaseri and on the ensheathing second-stage cuticle of H. bacteriophora did not reduce the nematodes' ability to infect larvae. The LD values for S. carpocapsae, S. glaseri, and H. bacteriophora in sand containing H. rhossiliensis were not different from those in sterilized sand when Galleria larvae were added at the same time as the nematodes. However, when Galleria larvae were added 3 days after the nematodes, the LD of S. glaseri was higher in Hirsutella-infested sand than in sterilized sand, whereas the LD of H. bacteriophora was the same in infested and sterilized sand. Although the LD of S. carpocapsae was much higher in Hirsutella-infested sand than in sterilized sand, the data were too variable to detect a significant difference. These data suggest that H. bacteriophora may be more effective than Steinernema species at reducing insect pests in habitats with abundant nematode-parasitic fungi.