Susceptibility of Sap Beetles (Coleoptera: Nitidulidae) to Entomopathogenic Nematodes

Nitidulid beetles (Coleoptera) are considered to be serious pests of date palms throughout the world. They attack ripe fruit, causing it to rot, and damage is reflected in both reduced yield and lower fruit quality. The present study was aimed at an evaluation of the susceptibility to different sap beetles to entomopathogenic nematodes. We further tested nematode efficacy in pots filled with soil infested by third instar larvae of the two beetle species. In Petri dish assay, mortality levels of Carpophilus humeralis and C. hemipterus exposed to Heterorhabditis sp. IS-5 strain indicated that the latter is less susceptible to nematode infection. Exposure of both sap beetle species to different nematode strains gave moderate levels of mortality (35-65%) with the heterorhabditid strains HP88, IS-5 and IS-25. The IS-12 strain of Heterorhabditis sp. showed poor virulence (<35% mortality) against larvae of C. humeralis as well as larvae and pupae of C. humipterus. The nematode species S. riobrave showed moderate virulence (35-65%) mortality to larvae and pupae of S. humeralis as well as to larvae of C. hemipterus . Exposure of C. hemeralis to different concentrations of Heterorhabditis sp. IS-5 in pots containing soil resulted in high mortality (>65%). In contrast, the lower concentrations (500 and 1000 nematodes/pot) caused low mortality (35%) of C. hemipterus . Other heterorhabditid strains caused 95-100% mortality of C. humeralis in pot assay. The HP88 strain of H. bacteriophora and the Tx strain of Steinernema riobrave showed poor effectiveness. Incubation of different nematode strains with the C. humeralis larvae at high temperature (32 C) resulted in an increase in insect mortality with the IS-12 and IS-21 strains. Reduced mortality was recorded with the HP88 strain treatment at the higher temperature. The IS-5 and IS-12 strains were equally effective in all three soil types tested, whereas the IS-19 strain was more effective in the Almog type soil than in the others.     

Efficacy and Persistence of Entomopathogenic Nematodes in Controlling Larval Populations of Thaumetopoea pityocampa (Lepidoptera: Thaumetopoeidae)

A 3-year study was conducted in a Pinus halepensis reforestation of Apulia Region (Southern Italy) injecting IJs (infective juveniles) of Steinernema feltiae , S. carpocapsae and Heterorhabditis bacteriophora in aqueous and gel suspensions (Idrosorb SR 2002 [Nigem ® ], and Compex) into the nests of Thaumetopoea pityocampa caterpillar. This study showed that the gel suspensions do not percolate and that slow release of water from the gels allowed nematodes to survive and complete their life cycle in the host. Results demonstrate the feasibility of reducing overwintering larval populations by injecting gel suspension of S. feltiae . We found no negative effects on the endoparasite Phryxe caudata .     

Suppression of Plant-parasitic Nematode Populations in Turfgrass by Application of Entomopathogenic Nematodes

We studied the influence of entomopathogenic nematodes , Steinernema carpocapsae and S. riobravis, on natural populations of plant - parasitic nematodes (PPNs) infesting turfgrass in Georgia and South Carolina . S. riobravis applied at 6 109 infective juveniles (IJs) / acre provided up to 95 - 100% control of the root - knot , Meloidogyne sp ., sting , Belonolaimus longicaudatus, and ring nematode , Criconemella sp ., in Georgia , but S. carpocapsae had no effect . S. riobravis was as effective as the chemical nematicide , Fenamiphos (Nemacur 10G) at 4 weeks after treatment and more effective at 8 weeks after treatment . In South Carolina , both S. riobravis and S. carpocapsae applied at 1 109 IJs / acre provided up to 86 - 100 % control of the root - knot , sting and ring nematodes . Application of 6 109 IJs / acre increased control by only 4 - 14 % over the 1 109 dosage . Possible causes of differences in efficacy of S. carpocapsae at the two sites are discussed . It is concluded that S. riobravis may provide effective , predictable and economical control of PPNs in turfgrass .     

Infectivity of Four Species of Nematodes (Rhabditoidea: Steinernematidae, Heterorhabditidae) to the Asian Longhorn Beetle, Anoplophora glabripennis (Motchulsky) (Coleoptera: Cerambycidae)

Four species of entomopathogenic nematodes, Steinernema carpocapsae , Heterorhabditis bacteriophora , H. indica and H. marelatus , were tested for their ability to kill and reproduce in larvae of the Asian longhorn beetle, Anoplophora glabripennis (Motchulsky). The larvae were permissive to all four species but mortality was higher and production of infective juveniles was greater for S. carpocapsae and H. marelatus . The lethal dosage of H. marelatus was determined to be 19 infective juveniles for second and third instar larvae and 347 infective juveniles for fourth and fifth instar larvae. H. marelatus infective juveniles, applied via sponges to oviposition sites on cut logs, located and killed host larvae within 30 cm galleries and reproduced successfully in several of the larvae.     

Evaluation of Entomopathogenic Nematodes for the Control of Mediterranean Fruit Fly (Diptera: Tephritidae)

The virulence of various entomopathogenic nematode (EPN) strains was evaluated against the Mediterranean fruit fly, C. capitata . The selected nematodes were assessed for their infectivity for the final larval stage of the insect host and under varying environmental conditions. Among 12 EPN strains tested, Steinernema riobrave Texas ( Sr TX) and Heterorhabditis sp. IS-5 (H IS-5), showed high activity and induced >80% mortality. Six EPN strains showed limited activity (>30% mortality), and four strains had no effect (<20% mortality). Sr TX was more effective than H IS-5. Mature C. capitata larvae were most susceptible to nematode infection during the first 4h after they began to emerge from their diet to pupate. Activity of the two nematode strains at a constant inoculation rate was dependent on insect larval density. The highest activity was recorded at 1.88 larvae cm -2 and decreased at higher larval densities. EPN activity was also directly related to nematode density. Maximal activity was shown at a density of 150 infective juveniles cm -2 . A similar activity pattern was also recorded with Sr TX in four different soil types. The persistence of this EPN in the soil extended over 5 days but there was no activity after 14 days. Except for a lower activity under cool conditions (17°C), temperatures ranging between 22 and 41°C, or moisture levels in the treated soil ranging between 3 and 20%, had no significant effect on nematode activity. Our results suggest that application of Sr TX against C. capitata may have potential for controlling C. capitata .     

Effect of Timber Condition on Parasitization of Pine Weevil ( Hylobius abietis L.) Larvae by Entomopathogenic Nematodes under Laboratory Conditions

The large pine weevil ( Hylobius abietis L.) is one of the most important pests in coniferous reforestation in Europe. Larvae develop in the stumps of recently felled trees; the emerging adults feed on the bark of seedlings and may kill them. The ability of the entomopathogenic nematodes Heterorhabditis megidis and Steinernema carpocapsae to invade pine weevil larvae in Sitka spruce ( Picea sitchensis ) buried in moist sand was evaluated. Overall, four times as many H. megidis as S. carpocapsae invaded pine weevil larvae. The two species of nematode differed in their response to timber condition. The number of S. carpocapsae invading pine weevil larvae was twice as high in billets inoculated with the wood-rotting fungus Phlebiopsis gigantea as in fresh timber, while the number of H. megidis invading was reduced by 25%. Invasion into non-feeding insects (larvae of the wax moth Galleria mellonella ) contained in timber disks was also affected by timber quality, indicating that nematode behaviour was affected directly by the physical or chemical condition of the timber, though trophically mediated effects may also have been involved.     

Attractant for the sugar-beet weevil Conorrhynchus (Cleonus) mendicus (Col.: Curculionidae)

Abstract:  Traps baited with a 1 : 1 : 1 mixture of Grandlure I [ cis -1-methyl-2-(1-methylenethenyl)-cyclobutaneethanol], Grandlure II [( Z )-2-ochtoden-1-ol; ( Z )-2-(3,3-dimethylcyclohexylidene)ethanol] and Grandlure III–IV [( Z )- and ( E )-2-ochtodenal; ( Z )- and ( E )-(3,3-dimethyl)cyclohexylidene)acetaldehyde] caught significantly more Conorrhynchus (Cleonus) mendicus Germar (Col.: Curculionidae) weevils than traps without bait. Later tests revealed that only Grandlure III–IV was responsible for attractivity. Both sexes were attracted according to a random sample of captures. C. mendicus appeared to be very similar in its responses to the synthetic aggregation attractant as the closely related Bothynoderes (Cleonus) punctiventris (Germar), in which species, the attraction to Grandluree III–IV has recently been published. Doses from 500 to 5000  μ g of Grandlure III–IV on rubber dispensers performed satisfactorily in monitoring trials and can be recommended for use in practical trapping applications for C. mendicus .     

Biological Control of the Pecan Weevil,Curculio caryae (Coleoptera: Curculionidae), with Entomopathogenic Nematodes

Steinernema carpocapsae (Weiser) strain A11, S. feltiae (Filipjev) strain SN, and Heterorhabditis bacteriophora Poinar strains HP88 and Georgia were tested for their efficacy as biological control agents of the pecan weevil, Curculio caryae (Horn), in pecan orchard soil-profile containers under greenhouse conditions. Percentage C. caryae parasitism by S. carpocapsae and H. bacteriophora strain HP88 and Georgia was consistently poor when applied either prior to or following C. caryae entry into the soil, suggesting that these nematode species and (or) their enterobacteria are poor biological control agents of weevil larvae. Soil taken 21 days following application of S. carpocapsae or H. bacteriophora strain HP88 induced a low rate of infection of Galleria mellonella larvae, whereas soil that had been similarily treated with H. bacteriophora strain Georgia induced a moderate rate of infection. Percentage C. caryae parasitism by S. feltiae was consistently low when applied following C. caryae entry into the soil and was inconsistent when applied as a barrier prior to entry of weevil larvae into the soil. Soil taken 21 days following application of S. feltiae induced a high rate of infection of G. mellonella larvae.     

Post-application Persistence of Entomopathogenic Nematodes

Five phases can be distinguished in the post-application persistence of entomopathogenic nematodes and each phase is associated with a specific set of mortality factors. Pre-application factors associated with production, storage and transport conditions determine the survival rate and quality of nematodes at the time of application. The phase of tank mixing and application with a sprayer, hose or other equipment does not usually cause mortality as nematode dauer juveniles are quite tolerant of shear forces. The most critical periods for survival are the first few minutes and hours directly after application. High losses, in the order of 40-80%, often occur during this phase. Ultraviolet radiation and dehydration are probably the most important mortality factors. The remaining nematodes settle in the soil and their numbers gradually decrease at levels of 5-10% per day. Predation, infection by antagonists, depletion of energy and desiccation are probably the main mortality factors during this period. In most cases, after 2-6 weeks less than 1% of the applied population is still alive. Through recycling in host insects, nematodes may then persist for years at these levels. Thus, the pattern is a rapid decline in the first few days followed by a moderate decline over the next 2-6 weeks and then a long period of recycling at a low level. Some nematode species that normally occur in warmer climatic zones can also persist in colder climates. Major side-effects of applications of entomopathogenic nematodes are not likely to occur as the population density decreases to background levels within days or weeks after application. Furthermore, there is little or no migration of the nematodes to neighbouring fields. The relatively short period of persistence of entomopathogenic nematodes and the necessity of their populations to recycle frequently in hosts in order to survive make it unlikely that they could have major effects on non-target organisms. Their selectivity and beneficial traits as biological control agents outweigh the small risks of causing unwanted environmental disturbance in non-target populations.     

Impact of Entomopathogenic Nematodes on Different Soil-dwelling Stages of Western Flower Thrips, Frankliniella occidentalis (Thysanoptera: Thripidae), in the Laboratory and Under Semi-field Conditions

The impact of entomopathogenic nematodes (EPN) on mortality of soil-dwelling stages of western flower thrips (WFT), Frankliniella occidentalis (Thysanoptera: Thripidae) with different insect stage combinations was studied in the laboratory and under semi-field conditions. In laboratory experiments, the efficacy of Steinernema feltiae strain Sylt (Rhabditida: Steinernematidae) at a concentration of 400 infective juveniles (IJs) cm -2 was tested against different proportions of soil-dwelling stages of WFT, i.e. late second instar larvae (L2), prepupae and pupae. Soil was used as the testing medium. S. feltiae significantly affected the mortality of all soil-dwelling life stages of WFT at all tested insect stage combinations. The proportion of late L2 in the population negatively correlated to EPN-induced mortality. WFT prepupa and pupa were similarly susceptible to S. feltiae and their proportion in the population did not affect the EPN-induced mortality under laboratory conditions. The highest mortality (80%) was recorded when the population consisted only of prepupae and/or pupae. In the semi-field study, the impact of S. feltiae , S. carpocapsae strain DD136 and Heterorhabditis bacteriophora strain HK3 (Rhabditida: Heterorhabditidae) ( H. bacteriophora ) at concentrations of 400 and 1000 IJs cm -2 was evaluated against WFT reared on green beans, Phaseolus vulgaris L., as host plant in pot experiments in a controlled climate chamber. All tested EPN strains at both dose rates significantly reduced the WFT populations. Up to 70% reduction of the WFT population was obtained at the higher EPN concentration.     

Genetic Enhancement of Nematicide Resistance in Entomopathogenic Nematodes

Entomopathogenic nematodes are highly effective bioinsecticides . Their efficacy may be reduced due to the various pesticides they encounter in the soil . These include insecticides as well as nematicides used against plant - parasitic nematodes . The purpose of this study was to examine the feasibility of genetic selection as a means of enhancing resistance of the entomopathogenic nematode Heterorhabditis bacteriophora strain HP88 to the nematicides: Fenamiphos (an organophosphate) , Oxamyl (a carbamate) and Avermectin (a biological product) . Estimates of heritability ( h 2) of resistance to the three nematicides were obtained from analysis of inbred lines derived from the base population . The heritability estimate for Fenamiphos was h 2 = 0 . 31 , for Oxamyl h 2 = 0 . 71 and for Avermectin h 2 = 0 . 46 . Five rounds of selection were performed . Thereafter , each line was divided into two: for one subline selection continued for six additional rounds . The other subline was reared without selection for the six additional rounds . After the eleventh round , resistance to the nematicides was examined as were several traits relevant to biocontrol efficacy including virulence , heat tolerance and reproduction potential . Selection resulted in an 8 - 9 - fold increase in resistance to Fenamiphos and Avermectin and a 70 - fold increase in resistance to Oxamyl . The enhanced resistance Oxamyl and Avermectin , and to a lesser extent to Fenamiphos , was stable and continued after selection was relaxed . No deterioration in traits relevant to biocontrol efficacy was observed in the selected lines as compared with the base population . The selected lines displayed enhanced cross - resistance towards some , but not all , of the nematicides tested . These results demonstrate that genetic selection can be used to enhance resistance of entomopatho genic nematodes to certain environmental stresses . The selected lines will be useful bioinsecti cides in the context of integrated pest management .     

EVect of the Entomopathogenic Nematodes, Steinernema feltiae and S. carpocapsae , on the Potato Cyst Nematode, Globodera rostochiensis , in Pot Trials

Two pot experiments, one in a glasshouse and the other in an outdoor sand plunge, were conducted to examine the influence of the entomopathogenic nematodes, Steinernema feltiae and S. carpocapsae , on the invasion and development of the potato cyst nematode, Globodera rostochiensis . Of a total of eight diVerent treatments with entomopathogenic nematodes in the glasshouse trial, three reduced the invasion of G. rostochiensis and one reduced the numbers of new cysts that were produced compared with controls. In the outdoor experiment, seven of the 12 treatments gave a reduction in invasion but none resulted in changes in the numbers of cysts found at plant senescence. In general, invasion of G. rostochiensis juveniles was reduced more eVectively by S. carpocapsae than by S. feltiae , and was greatest in the outdoor trial where larger inocula of entomopathogenic nematodes were used. Overall, the results indicated that use of S. feltiae and S. carpocapsae is unlikely to provide a viable control strategy for G. rostochiensis .     

Evaluation of Steinernema carpocapsae Weiser for Control of Carrot Weevil Adults, Listronotus oregonensis (LeConte) (Coleoptera: Curculionidae), in Organically Grown Carrots

Under controlled conditions, application of Steinernema carpocapsae using a baiting method consistently reduced the oviposition of Listronotus oregonensis adults at all moisture regimens. At 80 and 95% relative humidity, soil applications at 10 4 infective juveniles (IJs) cm-2 and 10 5 infective stages (400 IJs cm-2) per pot reduced the survival and oviposition of L. oregonensis adults; soil applications were ineffective at relative humidity lower than 80%. When timed correctly, soil application at the rate of 2 105 IJs/linear meter of row (4.4 109 IJs ha-1) reduced damage by up to 59%.     

Laboratory Trials of the Efficacy of the Crown Weevil Mortadelo horridus (Coleoptera: Curculionoidae) for the Biological Control of Slender Thistles Carduus pycnocephalus and C. tenuiflorus in Southwestern Australia

The hypothesis that larval crown weevil (Mortadelo horridus Alonso-Zarazaga and Sanchez-Ruiz) can suppress vegetative growth, seed set and viability in the winter annual slender thistles (Carduus pycnocephalus L. and C. tenuiflorus Curt.) was tested in laboratory gardens. Different densities (eight and 16) of M. horridus larvae and eggs were inoculated onto slender thistle rosettes between one and four times. Controls received no eggs or larvae. Larval feeding destroyed the apical meristems of treated plants, producing multi-crowned rosettes and significant declines in leaf area in both species, irrespective of the number of larvae. Treated thistles bolted at the same time and had a 4-fold increase in stem number compared to controls. The stems of control plants were up to one-third taller and thicker than attacked plants. Relative to controls, the leaves of treated plants had significantly fewer spines, decreased leaf area and decreased leaf perimeter, with less deeply lobed leaf margins. The total number of capitula produced by control and experimental groups was not significantly different, but consideration of fertile capitula only revealed a reduction in fertile capitula production following weevil attack. This was most marked in C. pycnocephalus. Control plants also produced significantly wider capitula. No control plants died, compared to 17 and 26% of the attacked plants of C. pycnocephalus and C. tenuiflorus, respectively. Implications for biocontrol of slender thistles are discussed.     

Enhanced Trehalose Accumulation and Desiccation Survival of Entomopathogenic Nematodes Through Cold Preacclimation

Limited storage stability is a major obstacle to further expansion of the use of entomopathogenic nematodes for pest control. Progress has been made that Steinernema carpocapsae can now be stored under partial anhydrobiosis for up to 6 months at 25°C and 10 months at 5°C in a water-dispersible granular (WG) formulation. However, other species have been more difficult to store in the WG formulation due to migration of nematodes out of the granules and sensitivity of some species to desiccation directly at cold temperatures. As acclimation to cold induces trehalose accumulation (a major cryo- and desiccation protectant) in many invertebrates, it was hypothesized that cold preacclimation of entomopathogenic nematodes will enhance their survival in the WG formulation at cold temperatures. This hypothesis was tested using a temperate species Steinernema feltiae , a subtropical species S. carpocapsae , and a tropical species Steinernema riobrave possessing different thermal niche breadths and reproduction temperature optima. Cold acclimation of infective juveniles increased trehalose accumulation in all three species and the amount of trehalose accumulated was both temperature and species dependent. Trehalose content reached at its peak after 6 days at 5°C in S. feltiae (82.28 μg/mg dry weight), after 10 days at 10°C in S. carpocapsae (94.16 μg/mg dry weight) and after 6 days at 15°C in S. riobrave (47.58 μg/mg dry weight). Cold preacclimation at 5°C for 2 days enhanced desiccation survival of S. feltiae in 25% glycerol (osmotic desiccation) at both 5 and 25° and of S. carpocapsae and S. riobrave only at 5°C. Non-cold acclimated S. carpocapsae and S. riobrave were extremely sensitive to desiccation directly at 5°C in 25% glycerol, resulting in over 98% mortality within 6 days, but S. feltiae was more sensitive to desiccation at 25°C than at 5°C. Cold preacclimation increased survival of all the three species in the WG formulation at both 5 and 25°C. The survival of S. riobrave at 5°C in the WG formulation was positively correlated with the length of preacclimation period at 5°C (R 2 = 0.99) and with the amount of trehalose accumulated during cold preacclimation (R 2 = 0.81). These results support the hypothesis that cold preacclimation enhances desiccation survival of entomopathogenic nematodes at cold temperatures and the increased survival correlates well with the increased trehalose accumulation. Results also demonstrate that cold preacclimation can be used as a tool to enhance survival of nematodes in the formulations with reduced water activity.     

Control of Black Vine Weevil larvae Otiorhynchus sulcatus (Fabricius) (Coleoptera:Curculionidae) in grow bags outdoors with nematodes

Abstract 1 Outdoor trials were carried out during 2001–02 on strawberries grown in commercial growing bags naturally infested with black vine weevil larvae (BVW) Otiorhynchus sulcatus in Co. Wexford, Ireland. 2 The two nematode isolates used in these trials were Heterorhabditis megidis (UK211) and Heterorhabditis downesi (K122), both laboratory cultured. Growing bags received nematodes either once (May 2001), twice (May and October 2001) or three times (May, October 2001 and May 2002). Ten days after each application date, nine blocks (of the total 27) were randomly selected, destructively assessed and discarded. 3 The single application (May 2001) resulted in a mortality of black vine weevil larvae, of 93.4% with H. megidis and 51.3% with H. downesi, compared with the control treatment at that date. Respective figures after the double application (May 2001 and October 2001) were 78.9 and 77.6% and after the triple application (May 2001, October 2001 and May 2002) the figures were 93.7 and 88.1%. 4 Results from these trials clearly indicate that entomopathogenic nematodes are good alternatives to chemical control of the black vine weevil on strawberries grown in growing bags in Ireland.     

Field efficacy of entomopathogenic nematodes against the beetle Maladera matrida (Coleoptera: Scarabaeidae)

Single, double and triple releases of the entomopathogenic nematode Heterorhabditis bacteriophora Poinar, reduced the population of the beetle Maladera matrida Argaman, infesting peanuts (’Shulamit’ cv.) by 70, 75 and 93% respectively in microplot tests. Simultaneous and late (2 weeks after infestation) applications reduced beetle numbers by 63 and 79% respectively, in the microplots, while early application (2 weeks prior to infestation) did not reduce the beetle population. In a field trial, reductions in insect population and damage to the crop were achieved by early treatment with the nematode as well as by Heptachlor, leading to reductions in the insect population of 60 and 90% respectively, when recorded 4 weeks after nematode application. However, the nematode treatment did not maintain its effectiveness for a longer period and pest damage increased to the same level as the untreated control after 7 weeks. When the nematodes were applied at different concentrations (0.25–1.0 x 10⁶ infective juveniles (IJs) m^‐2) their effectiveness was not related to the concentration level. The only significant (P < 0.05) reduction in insect levels was recorded in the treatment with 0.5 X 10⁶ IJs m^‐2. In a second field trial, both H. bacteriophora and Steinernema glaseri reduced insect populations significantly (P < 0.05) by approximately 50% in comparison to the control. In the third trial, treatment with H. bacteriophora resulted in a decrease in insect population of 90% while treatment with S. carpocapsae reduced the grub numbers by 40% in comparison to the control. A differential susceptibility of various grub developmental stages was recorded in the field. The small grubs (I‐4 mm long, lst‐2nd larval stage) were not affected by the nematode treatments while the numbers of medium and large size grubs were reduced by 2‐ and 3‐fold respectively in the various tests. Nematodes were recovered by ‘nematode traps’ containing Galleria mellonella larvae from treated field plots 78 days after application. The implications of the results from the present studies on the use of entomopathogenic nematodes are discussed in relation to the development of an integrated pest management programme.     

Entomopathogenic nematodes for control of Phyllophaga georgiana (Coleoptera: Scarabaeidae) in cranberries

A series of laboratory and greenhouse experiments evaluated the entomopathogenic nematodes Steinernema scarabaei Stock & Koppenhöfer, Heterorhabditis bacteriophora Poinar, and H. zealandica Poinar for control of second- and third-instar cranberry white grub, Phyllophaga georgiana Horn (Coleoptera: Scarabaeidae), in cranberries. Steinernema scarabaei was the most effective species with 76–100% control at a rate of 2.5×10⁹ IJ/ha in the greenhouse experiments. H. zealandica and especially H. bacteriophora were generally less effective and required rates of 5×10⁹ IJ/ha for acceptable control. Larval stage had no effect on H. zealandica and H. bacteriophora performance, whereas S. scarabaei was more effective against third instars than second instars in the laboratory but not in the greenhouse experiments. Steinernema scarabaei, should it become commercially available, could be an effective alternative to chemical insecticides for P. georgiana management.     

Biocontrol Potential of Entomopathogenic Nematodes Against Winter Moths (Operophtera brumata and O. fagata) (Lepidoptera: Geometridae) Infesting Urban Trees

Infectivity and biocontrol potential of entomopathogenic nematodes against winter moths (Operophtera brumata and O. fagata)pupating in the soil were examined in laboratory, semi-field and field conditions. A pilot experiment conducted in the field showed that Steinernema feltiae was completely ineffective against pupae of these moths in the soil. Subsequent laboratory tests revealed that none of the tested species (i.e. S. feltiae, S. affinae, S. carpocapsae, Heterorhabditis megidis and H. bacteriophora) could colonise the pupae, while mature larvae descending to the soil for pupation and prepupae were highly susceptible to nematode infection. No differences were observed between O. brumata and O. fagata in susceptibility to nematodes. In laboratory experiments H. megidis applied at 1.5×10⁵infective juveniles (IJ) m^-2infected almost 100% of insects exposed for 6 days in the soil. It was significantly more infective than H. bacteriophora (73-77%) and Steinernema species (29-50%). H. megidis was also highly effective in semi-field conditions when applied at an even lower dose, i.e. 10⁵IJ m^-2. After a 45-day experiment, only 3% of insects descending for pupation survived in the soil pre-treated with this species. This was significantly less than in soil with S. feltiae (43%) and control treated with water only (59%). Very high efficacy of H. megidis and a relatively easy method for its field application through ground spraying gives some promise for environmentally safe and successful biological control of winter moths during their pupation in the soil. The low application rate required and recycling in the host could be additional advantages for economic and long lasting protection of high value trees, particularly those in urban parks and forests.     

Entomopathogenic Nematode Infectivity Assay: Comparison of Penetration Rate into Different Hosts

Penetration rate (the percentage of the initial infective juvenile inoculum that invades an insect host) was tested as an indicator of entomopathogenic nematode infectivity. Several host-parasite-substrate combinations were evaluated for penetration rate. Four steinernematids, Steinernema carpocapsae, S. glaseri, S. feltiae, S. riobravis and two strains of Heterorhabditis bacteriophora were tested in a contact bioassay against the wax moth, Galleria mellonella, the yellow meal worm, Tenebrio molitor, the beet armyworm, Spodoptera exigua, the black cutworm, Agrotis ipsilon, and the European corn borer, Ostrinia nubilalis. The insect larvae were confined individually in sand and filter paper arenas and exposed to 200 infective juveniles. After incubation, dead insects were dissected in order to count the nematodes penetrated. The data were analyzed for the effects of nematode strain and substrate on penetration rate. The bioassay substrate had a variable effect depending on the insect species. The nematode effect was highly significant for all insects tested. The penetration rate therefore allowed comparisons among nematode strains invading a host. Nematode ranking for infectivity differed according to the insect tested.