Survival and Movement of Insect Parasitic Nematodes in Poultry Manure and Their Infectivity Against Musca domestica

Survival, infectivity, and movement of three insect parasitic nematodes (Steinernema feltiae All strain, S. bibionis SN strain, and Heterorhabditis heliothidis NC strain) in poultry manure were tested under laboratory conditions. The majority (70-100%) of the nematodes died within 18 hours after exposure to the manure. Nematodes exposed to manure slurry for 6 hours killed at least 95% of the house fly larvae, Musca domestica, but nematodes exposed for 12 hours achieved less than 40% larval mortality. The majority (90-97%) of the three nematode species applied to the manure remained on the surface. Poor survival and limited movement of nematodes in poultry manure appear to make them unlikely candidates for biocontrol of filth flies in this habitat.     

Soil Applications of Steinernematid and Heterorhabditid Nematodes for Control of Colorado Potato Beetles,Leptinotarsa decemlineata (Say)

Three strains of Steinernema feltiae Filipjev (All, Mexican, and Breton strains) and one of Heterorhabditis heliothidis (Khan, Brooks, and Hirschmann) were evaluated for their potential to control Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), larvae and pupae in the soil. In laboratory studies, H. heliothidis and S. feltiae (Mexican strain) produced the highest mortality (6 days posttreatment) of CPB when applied to the surface of a soil column containing mature CPB larvae 5 cm below. Mortality ranged from 80 to 90% at rates of 79-158 nematodes/cm². Similar results were seen in a field microplot study with all four nematodes; S. feltiae (Mexican strain) and H. heliothidis were most effective. Adult CPB emergence was reduced 86.5-100% after application of 31-93 H. heliothidis/cm² and 88.4-100% with 93-155 S. feltiae (Mexican strain)/cm². The All strain of S. feltiae was moderately effective (ca. 80% reduction at 93-155 nematodes/cm²), while the Breton strain was ineffective (< 40% reduction at 155 nematodes/cm²). In small plots of potatoes enclosed in field cages, application of H. heliothidis and S. feltiae (Mexican strain) at rates of 93-155 nematodes/cm² before larval CPB burial in the soil resulted in 66-77% reduction in adult CPB emergence. Soil applications of these nematodes show potential for biological control of CPB.     

Susceptibility of the Carrot Weevil (Coleoptera: Curculionidae) to Steinernema feltiae,S. bibionis,and Heterorhabditis heliothidis

Larvae, pupae, and adults of the carrot weevil (Listronotus oregonensis) were infected and killed by the three entomophagous nematodes (Steinernema feltiae, S. bibionis, and Heterorhabditis heliothidis) under controlled conditions. Third-stage larvae were more susceptible than pupae or adults. S. feltiae and S. bibionis were the most aggressive nematode species, causing larval mortality after 24-48 hours in both continuous and 2-hour contact with nematode suspension. The nematodes multiplied sufficiently in all insects at all stages of development; however, production of infective-stage larvae per host cadaver was variable.     

Susceptibility of Meligethes spp. and Dasyneura brassicae to entomopathogenic nematodes during pupation in soil

The susceptibility of pupating larvae of pollen beetles, Meligethes spp. Stephens (Coleoptera: Nitidulidae) and brassica pod midges, Dasyneura brassicae Winnertz (Diptera: Cecidomyidae) to entomopathogenic nematodes (Nematoda: Rhabditida) was studied in the laboratory. The results showed that brassica pod midge larvae were almost unaffected by the tested nematodes (Steinernema bicornutum, S. feltiae and Heterorhabditis bacteriophora) whereas successful pupation of pollen beetle larvae was reduced with increasing number of nematodes (S. bicornutum, S. carpocapsae, S. feltiae and H. bacteriophora). The exposed larvae had been collected in the field and some of the pollen beetle larvae were parasitised by parasitoid wasps. It appeared that parasitised larvae were less affected by nematodes than non-parasitised larvae.     

Entomopathogenic nematodes control Plum Sawflies (Hoplocampa minuta and H. flava)

Plum sawflies are among the most damaging pests of European plum. Current control strategy implies insecticide application. Three species of entomopathogenic nematodes (EPN), Steinernema feltiae Filipjev, S. carpocapsae Weiser and Heterorhabditis bacteriophora Poinar were tested under laboratory and field conditions to assess effectiveness against larval and adult stages. Laboratory tests resulted in up to 100% mortality of last instar larvae before construction of a cocoon. However, the nematodes were not able to penetrate the cocoon. Foliar application did not result in plum sawflies larvae infestation by EPNs. Under field conditions, the nematodes reduced the number of emerging adults by application against sawfly larvae in the previous year before migration into the soil for overwintering by 62%–92%. Application of the nematodes against adults just before their anticipated emergence resulted in reduction of fruit infestation up to 100%. Mean results of 5 trials using caged trees were 47.8% with S. feltiae, 56.3% with S. carpocapsae and 62.9% with H. bacteriophora. In open field trails, control of adults obtained with S. feltiae at 0.5 million nematodes/m² was 98.2 and 67.8% and at 0.25 million m⁻² 41.7 and 41.2%. Forecasting adult emergence and optimal soil moisture conditions are essential for success of the nematode application.     

Susceptibility of Delia radicum to steinernematid nematodes

Isolates of different Steinernema species (S. affine, S. bicornutum, S. feltiae and Steinernema C1) were used in mortality assays with third instar larvae of Delia radicum (L.) (Diptera: Anthomyiidae). The nematode isolates had been obtained by baiting soil regularly grown with cabbage. One isolate (S. feltiae) was the result of a natural infection of a D. radicum puparium. The highest mortality (77%) was obtained with an isolate of S. feltiae (DK1). The isolate DK1 was also used in tests with all larval stages of D. radicum. Mortality around 60% was observed for second and third instar larvae, while first instar larvae showed very low or no susceptibility. Maximum mortality of second and third instar larvae was reached applying only 25 nematodes per larva. Observations of larvae that pupated revealed that some of these puparia contained nematodes. Experiments with hatching puparia showed that a high proportion was infected by nematodes if the flies were prevented from leaving nematode-containing soil. In addition to mortality, the ability of the nematodes to successfully reproduce in the insects was studied. It was found that the species S. feltiae and S. bicornutum reproduced in D. radicum larvae and adults with S. feltiae being the most successful.     

Effectiveness of entomopathogenic nematodes Steinernema feltiae and Heterorhabditis bacteriophora on the Colorado potato beetle Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae) under laboratory and greenhouse conditions

In laboratory and greenhouse studies, the invading ability, virulence, and mortality caused by Stinernema feltiae and Heterorhabditis bacteriophora were compared. After one and two days of exposure to either nematode species, the mortality of Colordo potato beetle (CPB) Leptinotarsa decemlineata larvae at different instars, third and fourth, was recorded and the number of nematodes invading cadavers was more than the number of nematodes inside the larvae at the late last instar (one day before pre-pupa). Two concentrations, 250 and 500 IJs/dish, infective juvenile nematodes/0.5 ml were tested on different CPB larval instar. S. feltiae was more effective, with fourth instar rather than third and late last instar. On the other hand, H. bacteriophora showed a very weak effect with L. decemlineata. Also it was clear that S. feltiae was more effective and faster than H. bacteriophora: more than 70% of larvae were killed within 24 hours compared with H. bacteriophora which killed 40% of larvae within 48–72 hours. A significant difference in invading efficiency was observed with concentration 2500 IJs/pot in the greenhouse test. The number of adult females found in the cadavers of L. decemlineata larvae was always higher than the number of males. Foliage application of S. feltiae and H. bacteriophora resulted in a significant reduction of the number of damaged leaves and a lower index of damage compared with that in the control. We conclude that S. feltiae has significant potential and can help in the management of the Colorado potato beetle.     

Susceptibility of the Colorado Potato Beetle and the Sugarbeet Wireworm to Steinernema feltiae and S. glaseri

In laboratory tests, larvae of the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), and the sugarbeet wireworm (SBW), Limonius californicus (Mannerheim), were exposed to the nematodes Steinernema feltiae Filipjev (Mexican strain) (= Neoaplectana carpocapsae) and S. glaseri Steiner in soil. S. feltiae caused significantly higher mortality in SBW larvae than did S. glaseri, but both nematode species were equally effective against CPB larvae. The minimum concentration of S. feltiae for 100% mortality of CPB larvae after 13 days was 157 nematodes/cm² of soil, and the LC₅₀ based on 6-day mortality was 47.5 nematodes/cm²; in contrast, 100% mortality of SBW larvae was not achieved with even the highest concentration tested, 393 nematodes/cm². CPB adults emerging from nematode-contaminated soil were not infected. In field cage tests, S. feltiae applied to the soil surface at the rates of 155 and 310 nematodes/cm² soil caused 59% and 71% mortality, respectively, of late-fourth-instar spring-generation CPB, and 28% and 29% mortality, respectively, of SBW. No infection was obtained when larvae of summer generation CPB and SBW were placed in the same cages approximately 6 weeks after nematodes were applied to the soil. Inundative soil applications of S. feltiae, though cost prohibitive at present, were effective in reducing caged CPB and SBW field populations.     

Effect of temperature on the development of <Emphasis Type="Italic">Steinernema carpocapsae</Emphasis> and <Emphasis Type="Italic">Steinernema feltiae</Emphasis> (Nematoda: Rhabditida) in liquid culture

For commercial use of the entomopathogenic nematodes Steinernema carpocapsae and Steinernema feltiae in biological control of insect pests, they are produced in liquid culture on artificial media pre-incubated with their symbiotic bacteria Xenorhabdus nematophila and Xenorhabdus bovienii, respectively. After 1 day of the bacterial culture, nematode dauer juveniles (DJs) are inoculated, which recover development. The adult nematodes produce DJ offspring, which are harvested and can be sprayed. This study determined optimal temperatures to obtain high DJ progeny within a short process time. Temperatures assessed were 23°C, 25°C, 27°C, and 29°C for S. carpocapsae and 20°C, 23°C, 25°C, and 27°C for S. feltiae. The recovery of inoculated DJs was hardly affected and was reduced only in S. carpocapsae at 29°C. The fecundity (eggs in uterus) in S. carpocapsae reached a maximum at 27°C; whereas, maximum yields were recorded at 25°C. For both Steinernema spp., highest DJ densities were obtained after 15 days incubation at 25°C. Optimal culture temperature for both nematode species is 25°C. S. carpocapsae was more sensible to suboptimal temperature than S. feltiae. Results on total DJ density and DJ proportion of the total nematode population were more variable at non-optimal temperature condition for S. carpocapsae than for S. feltiae. Suboptimal culture temperature also reduced DJ infectivity.     

Susceptibility of the Plum Curculio,Conotrachelus nenuphar,to Entomopathogenic Nematodes

The plum curculio, Conotrachelus nenuphar, is a major pest of pome and stone fruit. Our objective was to determine virulence and reproductive potential of six commercially available nematode species in C. nenuphar larvae and adults. Nematodes tested were Heterorhabditis bacteriophora (Hb strain), H. marelatus (Point Reyes strains), H. megidis (UK211 strain), Steinernema riobrave (355 strain), S. carpocapsae (All strain), and S. feltiae (SN strain). Survival of C. nenuphar larvae treated with S. feltiae and S. riobrave, and survival of adults treated with S. carpocapsae and S. riobrave, was reduced relative to non-treated insects. Other nematode treatments were not different from the control. Conotrachelus nenuphar larvae were more susceptible to S. feltiae infection than were adults, but for other nematode species there was no significant insect-stage effect. Reproduction in C. nenuphar was greatest for H. marelatus, which produced approximately 10,000 nematodes in larvae and 5,500 in adults. Other nematodes produced approximately 1,000 to 3,700 infective juveniles per C. nenuphar with no significant differences among nematode species or insect stages. We conclude that S. carpocapsae or S. riobrave appears to have the most potential for controlling adults, whereas S. feltiae or S. riobrave appears to have the most potential for larval control.     

Efficacy and persistence of entomopathogenic nematodes for black cutworm control in turfgrass

Field experiments were conducted in turf maintained under golf course fairway conditions in May, June, and August 2009 and in August and September 2010 to evaluate the ability of entomopathogenic nematodes to control larval populations of the black cutworm, Agrotis ipsilon, on golf courses. Commercial products containing the entomopathogenic nematodes Heterorhabditis bacteriophora, Steinernema carpocapsae, S. feltiae, and S. riobrave were applied at 1.0 or 2.5×10⁹ infective juveniles per ha against fourth-instar black cutworms. Larval mortality and turf damage were evaluated at 4 and/or 7 days after treatment (DAT). Steinernema carpocapsae was the best performing species due to a combination of high control rates (average 83%), most consistent results (70–90% range), high speed of kill (average 68% at 4 DAT), and prevention of significant turf damage despite very high larval densities at 0 DAT. Efficacy of S. feltiae and H. bacteriophora was often similar to that of S. carpocapsae but overall less consistent. Short-term persistence of the nematodes was evaluated in four turfgrass sites maintained under golf course putting green, fairway, or rough conditions in June and August 2009 by baiting soil samples at 0, 4, 7, and 14 DAT. Relative to recovery immediately after application, at least 50% of S. feltiae and 25% of S. carpocapsae consistently persisted up to 4 days in one of the greens and up to 7 days in some trials. Our finding suggests that S. carpocapsae and S. feltiae may provide adequate black cutworm control in golf course turf under moderate summer temperatures.     

Stage-specific mortality of Rhagoletis indifferens (Diptera: Tephritidae) exposed to three species of Steinernema nematodes

Mortality of larval, pupal, and adult western cherry fruit fly, Rhagoletis indifferens (Tephritidae) exposed to the steinernematid nematodes Steinernema carpocapsae, Steinernema feltiae, and Steinernema intermedium, was determined in the laboratory and field. Larvae were the most susceptible stage, with mortality in the three nematode treatments ranging from 62 to 100%. S. carpocapsae and S. feltiae were equally effective against larvae at both 50 and 100 infective juveniles (IJs)/cm². S. intermedium was slightly less effective against larvae than the other two species. Mortalities of R. indifferens larvae at 0, 2, 4, and 6 days following their introduction into soil previously treated with S. carpocapsae and S. feltiae at 50 IJs/cm² were 78.6, 92.5, 95.0, and 77.5% and 87.5, 52.5, 92.5, and 70.0%, respectively, and at 100 IJs/cm² were 90.0, 92.0, 100.0, and 84.0% and 90.0, 50.0, 42.0, and 40.0%, respectively. There was no decline in mortality caused by S. carpocapsae as time progressed, whereas there was in one test with S. feltiae. Larval mortalities caused by the two species were the same in a 1:1:1 vermiculite:peat moss:sand soil mix and a more compact silt loam soil. In the field, S. carpocapsae and S. feltiae were equally effective against larvae. Pupae were not infected, but adult flies were infected by all three nematode species in the laboratory. S. carpocapsae was the most effective species at a concentration of 100 IJs/cm² and infected 11–53% of adults that emerged. The high pathogenicity of S. carpocapsae and S. feltiae against R. indifferens larvae and their persistence in soil as well as efficacy in different soil types indicate both nematodes hold promise as effective biological control agents of flies in isolated and abandoned lots or in yards of homeowners.     

Occurrence of Steinernema species in cabbage fields and the effect of inoculated S. feltiae on Delia radicum and its parasitoids

Abstract 1 Seven organically grown cabbage fields were surveyed for entomopathogenic nematodes in the autumn by baiting. Nematodes were obtained from three fields with bait larvae infection ranging from 1.3–4.0%. 2 Inoculation of Steinernema feltiae (Rhabditida: Steinernematidae) in the spring increased bait larvae infection to 16.0–32.0%. 3 Four different species were found (Steinernema affine, Steinernema bicornutum, S. feltiae and Steinernema C1). 4 The number of Delia radicum (L.) (Diptera: Anthomyiidae) puparia was significantly reduced at plants where S. feltiae had been inoculated. 5 Delia radicum parasitoids were also affected by S. feltiae. The results indicated that Aleochara species (Coleoptera: Staphylinidae) were more negatively affected than Trypliographa rapae (Westw.) (Hymenoptera: Cynipidae).     

Control of Larval Northern Corn Rootworm. (Diabrotica barberi) with Two Steinernematid Nematode Species

The entomogenous nematodes Steinerema feltiae and S. bibionis did not penetrate the roots of corn, Zea mays, to infect larval northern corn rootworm (NCR), Diabrotica barberi, feeding within. Laboratory bioassays against first instar NCR indicated that S. feltiae, Mexican strain (LD₅₀ = 49 nematodes/insect) is more virulent than S. bibionis (LD₅₀ = 100). Numbers of NCR larvae in a grain corn crop were reduced by both nematode species applied at corn seeding time at the rate of 10,000 infective-stage juveniles per linear meter of corn row. The chemical insecticide fonofos provided significantly better control than either nematode species.     

Infectivity of Steinernema carpocapsae and S. feltiae to Larvae and Adults of the Hazelnut Weevil,Curculio nucum: Differential Virulence and Entry Routes

We investigated the existing susceptibility differences of the hazelnut weevil, Curculio nucum L. (Coleoptera:, Curculionidae) to entomopathogenic nematodes by assessing the main route of entry of the nematodes, Steinernema carpocapsae strain B14 and S. feltiae strain D114, into larvae and adult insects, as well as host immune response. Our results suggested that S. carpocapsae B14 and S. feltiae D114 primarily entered adult insects and larvae through the anus. Larvae were more susceptible to S. feltiae D114 than S. carpocapsae B14 and adults were highly susceptible to S. carpocapsae B14 but displayed low susceptibility to S. feltiae D114. Penetration rate correlated with nematode virulence. We observed little evidence that hazelnut weevils mounted any cellular immune response toward S. carpocapsae B14 or S. feltiae D114. We conclude the differential susceptibility of hazelnut weevil larvae and adults to S. carpocapsae B14 and S. feltiae D114 primarily reflected differences in the ability of these two nematodes to penetrate the host.     

Efficacy of entomopathogenic nematodes against potato tuber moth,Phthorimaea operculella (Lepidoptera: Gelechiidae) under laboratory conditions

The susceptibility of potato tuber moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae) to native and commercial strains of entomopathogenic nematodes (EPNs) was studied under laboratory conditions. Native strains of EPNs were collected from northeastern Iran and characterised as Steinernema feltiae and Heterorhabditis bacteriophora (FUM 7) using classic methods as well as analysis of internal transcribed spacer (ITS) and D2/D3 sequences of 28S genes. Plate assays were performed to evaluate the efficiency of five EPN strains belonging to four species including Steinernema carpocapsae (commercial strain), S. feltiae, Steinernem glaseri and H. bacteriophora (FUM 7 and commercial strains). This initial assessment with 0, 75, 150, 250, 375 and 500 IJs/ml concentrations showed that S. carpocapsae and H. bacteriophora caused the highest mortality in both larval and prepupal stages of P. operculella, PTM. Thereafter, these three strains (i.e. S. carpocapsae, H. bacteriophora FUM 7 and the commercial strains) were selected for complementary assays to determine the effects of soil type (loamy, loamy–sandy and sandy) on the virulence of EPNs against the second (L2) and fourth instar (L4) larvae as well as prepupa. A soil column assay was conducted using 500 and 2000 IJs in 2-ml distilled water. Mortality in the L2 larvae was not affected by the EPN strain or soil type, while there was a significant interactive effect of nematode strains and soil type on larval mortality. The results also showed that EPN strains have higher efficiency in lighter soils and caused higher mortality on early larvae than that in loamy soil. In L4 larvae, mortality of PTM was significantly influenced by nematode strain and applied concentrations of infective juveniles. The larval mortality induced by S. carpocapsae was higher than those caused either by a commercial or the FUM 7 strain of H. bacteriophora. Prepupa were the most susceptible stage.     

Effect of Host Age and Nematode Strain on Susceptibility of Spodoptera frugiperda to Steinernema feltiae

Median lethal concentrations (LC₅₀) were determined for four nematode populations (two strains of Steinernema feltiae, a S. feltiae hybrid, and S. bibionis) against fifth-instar fall armyworm (Spodoptera frugiperda) larvae and for the most virulent of these nematodes against different instars and stages of the insect. Based on lack of overlap of 95% fiducial limits, there were significant differences in virulence among the four nematodes. The LC₅₀ ranged from 7.6 to 33.3 nematodes/ 0.7 ml water, and slopes of the log dose-probit regression lines were similar except for the S. feltiae All strain. First-instar fall armyworms suffered virtually 100% mortality from the S. feltiae Mexican strain at 1.0 nematode/0.7 ml, and LC₅₀ were 2.3 and 7.9 nematodes/0.7 ml in third-instar and fifth-instar larvae, respectively. Pupae had 7-20% mortality at doses ranging from 30 to 60 nematodes/0.7 ml.     

Symbiont-mediated competition: Xenorhabdus bovienii confer an advantage to their nematode host Steinernema affine by killing competitor Steinernema feltiae

Bacterial symbionts can affect several biotic interactions of their hosts, including their competition with other species. Nematodes in the genus Steinernema utilize Xenorhabdus bacterial symbionts for insect host killing and nutritional bioconversion. Here, we establish that the Xenorhabdus bovienii bacterial symbiont (Xb-Sa-78) of Steinernema affine nematodes can impact competition between S. affine and S. feltiae by a novel mechanism, directly attacking its nematode competitor. Through co-injection and natural infection assays we demonstrate the causal role of Xb-Sa-78 in the superiority of S. affine over S. feltiae nematodes during competition. Survival assays revealed that Xb-Sa-78 bacteria kill reproductive life stages of S. feltiae. Microscopy and timed infection assays indicate that Xb-Sa-78 bacteria colonize S. feltiae nematode intestines, which alters morphology of the intestine. These data suggest that Xb-Sa-78 may be an intestinal pathogen of the non-native S. feltiae nematode, although it is a nonharmful colonizer of the native nematode host, S. affine. Screening additional X. bovienii isolates revealed that intestinal infection and killing of S. feltiae is conserved among isolates from nematodes closely related to S. affine, although the underlying killing mechanisms may vary. Together, these data demonstrate that bacterial symbionts can modulate competition between their hosts, and reinforce specificity in mutualistic interactions.     

Safety evaluation of four entomopathogenic nematode species against two silkworm species

Chinese oak silkworm (Antheraea pernyi) and mulberry silkworm (Bombyx mori) are economically important insects used for silk production and food resource. Entomopathogenic nematodes (EPNs) from the families of Steinernematidae and Heterorhabditidae are beneficial organisms currently considered in biological control. In this paper, we evaluated survival of two silkworm species exposed to four Steinernema species which are widely applied in pest control. The results showed that among four Steinernema species, S. bicornutum and S. feltiae did not have an effect on the larval survival to the two silkworm species, whereas S. carpocapsae and S. glaseri did have an effect. Each Steinernema species poses no threat to hatchability of eggs, pupation rate, larval durations and cocoon shell ratio.     

Predation of entomopathogenic nematodes by <Emphasis Type="Italic">Sancassania</Emphasis> sp. (Acari: Acaridae)

Predation of the entomopathogenic nematode, Steinernema feltiae (Rhabditida: Steinernematidae), by Sancassania sp. (Acari: Acaridae) isolated from field-collected scarab larvae was examined under laboratory conditions. Adult female mites consumed more than 80% of the infective juvenile (IJ) stage of S. feltiae within 24 h. When S. feltiae IJs were exposed to the mites for 24 h and then exposed to Galleria mellonella (Lepidoptera: Pyralidae) larvae, the number of nematodes penetrating into the larvae was significantly lower compared to S. feltiae IJs that were not exposed to mites (control). Soil type significantly affected the predation rate of IJs by the mites. Mites preyed more on nematodes in sandy soil than in loamy soil. We also observed that the mites consumed more S. feltiae IJs than Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae). No phoretic relationship was observed between mites and nematodes and the nematodes did not infect the mites.