Susceptibility of larvae of the sheep blowfly Lucilia cuprina to various Heterorhabditis spp., Neoaplectana spp., and in undescribed steinernematid (Nematoda)

The susceptibility of third state larvae of the sheep blowfly Lucilia cuprina in sand to 11 species and strains of Heterorhabditis and Neoaplectana and to one species of an undescribed steiner-nematid was tested at various dosages. Larvae were susceptible to all, although far less so to one strain of N. bibionis and to the undescribed steinernematid. Estimates of LD₅₀ and LD₉₀ are presented for each effective strain. Heterorhabditis spp. were able to reproduce in L. cuprina larvae subjected to low dosages of effective-stage juveniles whereas Neoaplectana spp. were not able to reproduce at any dosage. The possibility exists for using Heterorhabditis species as a control agent against L. cuprina larvae after these have left the sheep to pupate in the soil.     

Comparative assessment of the efficacy of entomopathogenic nematode species at reducing western corn rootworm larvae and root damage in maize

The western corn rootworm (Diabrotica virgifera virgifera LeConte, Coleoptera: Chrysomelidae) is an invasive maize (Zea mays L.) pest in Europe. Crop yield is significantly impacted by the feeding of all three larval instars on maize roots, making them prime targets for control measures. Therefore, the control efficacy of three entomopathogenic nematodes (EPNs), Steinernema feltiae (Filipjev), Heterorhabditis bacteriophora Poinar, and H. megidis Poinar, Jackson and Klein (Nematoda: Rhabditida), was studied in four field plot experiments in southern Hungary in 2005 and 2006. All EPN species significantly reduced D. v. virgifera independently, whether applied as a row spray with a solid stream into the soil at sowing or onto the soil along maize rows in June. When applied at maize sowing, H. bacteriophora was more effective at reducing D. v. virgifera (81%) than H. megidis (49%) and S. feltiae (36%). When applied in June, H. bacteriophora and H. megidis were more effective at reducing D. v. virgifera (around 70%) than S. feltiae (32%). All tested EPN species significantly reduced damage on maize roots independently, whether they were applied at sowing or in June. Damage, however, was not totally prevented. The use of H. bacteriophora for the development of a biological control product for inundative releases against D. v. virgifera larvae is suggested.     

Assessment of establishment and persistence of entomopathogenic nematodes for biological control of western corn rootworm

Abstract:  The use of entomopathogenic nematodes (EPN) is potentially one ecological approach to control the invasive alien western corn rootworm ( Diabrotica virgifera virgifera LeConte, Col., Chrysomelidae) in Europe. This study investigated the establishment and the short- and long-term persistence of Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae), Heterorhabditis megidis Poinar, Jackson and Klein (Rh., Heterorhabditidae) and Steinernema feltiae Filipjev (Rh., Steinernematidae) in three maize fields in southern Hungary, using the insect-baiting technique. All three EPN species equally established and persisted in maize fields. The timing of application (April or June) did not influence the establishment of EPN species. EPNs persisted for 2–5 months, i.e. they survived up to and throughout D. v. virgifera larval occurrence in the soil. Results demonstrate that D. v. virgifera larvae can potentially be controlled by EPNs during the same year of EPN application but no long-term control effect is expected under intensive maize cultivation practices.     

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.     

ADVANCES IN THE RESEARCH OF ENTOMOPATHOGENIC NEMATODES STEINERNEMA AND HETERORHABDITIS IN CHINA

Abstract  The current status of Steinernema and Heterorhabditis entornopathogenic nematode research in China was reviewed in the collection and identification, the application in pest control programmes, symbiotic bacteria and mass production.     

Dispersal Behaviour and Transmission Strategies of the Entomopathogenic Nematodes Heterorhabditis and Steinernema

Entomopathogenic nematodes of the Heterorhabditidae and Steinernematidae appear to be capable of long-distance dispersal and local migration. Their transmission strategies include both highly active seek-and-destroy behaviours and ambusher strategies, and they may be sensitive to sex-related factors in their own populations. Their host-finding abilities are poorly understood, despite the fact that these abilities are fundamental to their success as biocontrol agents in soil. Like the vast numbers of exotic hymenopterans and other natural enemies that have been released for biological control over the past century, they may be used in their ecologically competent wild-type form. On the other hand, because they are applied inundatively, they may be tailored, by breeding or transformation, to their intended purpose and to ecological incompetence, improving both their efficacy and their ecological safety.     

中国昆虫病原斯氏和异小杆线虫的研究概况(英语)

昆虫病原斯氏和导小杆线虫作为生物防治因子已受到世界各国的广泛重视,并得到了商品化的发展.自50年代,特别是7O年代末,我国从原捷克斯洛伐克、澳大利亚、美国等地引进了大批昆虫病原斯氏和异小杆线虫.同时也从土壤或自然寄主中分离出许多本国线虫种或品系,并对此类线虫进行了全面的研究,在昆虫病原线虫的种类调查和鉴定、田间大面积应用、安全性、共生细菌、大量繁殖等方面取得了可喜的进展.本文将综述这些方面的研究概况.     

Field Efficacy of Entomopathogenic Nematodes against the Sugar-beet Weevil Temnorhinus ( = Conorrhynchus ) mendicus Gyll. (Coleoptera: Curculionidae)

In 1992 and 1993, the field effectiveness of Heterorhabditis sp. (NL-HL81 strain), H. bacteriophora (HP 88 strain) and Steinernema carpocapsae ('All' strain) against the larvae of Temnorhinus mendicus Gyll. was assessed. The biological tests were compared with two chemical treatments (cypermethrin or deltamethrin) and one untreated control. In 1992, S. carpocapsae gave better results than Heterorhabditis sp. in reducing the percentage of infested roots, as compared with the untreated sample and the chemical one; similarly, the irrigated control gave the best results. In 1993, three concentrations of entomopathogenic nematodes (EPNs) were tested: 0.250 106 infective juveniles (IJs) m - 2, 0.125 106 IJs m - 2 and 0.075 106 IJs m - 2. The different numbers of EPNs did not give very different results from each other; however, H. bacteriophora at 0.075 106 IJs m - 2 was the least effective. In general, cypermethrin was more effective than deltamethrin, but one treatment with EPNs followed by irrigation was always more effective than two chemical applications.     

Synergistic effect of the herbicides glyphosate and MCPA on survival of entomopathogenic nematodes

The survival and infectivity of the infective juveniles of two species of entomopathogenic nematodes, Steinernema feltiae (Rhabditida: Steinernematidae) Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae), were determined after exposure for 72 h to two concentrations of the herbicides glyphosate and MCPA, as well as to the combination of the two herbicides (glyphosate + MCPA). For all herbicide treatments, concentrations and exposure times, S. feltiae was more tolerant to the herbicides than H. bacteriophora. The exposure of entomopathogenic nematodes to glyphosate + MCPA caused significantly higher mortality (26.33–57.33%) than glyphosate (0.67–15%) or MCPA (2.33–19%) alone. These results confirm the synergistic effect of the glyphosate + MCPA combination on the mortality in these nematodes. Nematode infectivity of Galleria mellonella larvae in response to the herbicides presence was evaluated in Petri dish assays containing sterile sand. Nematode infectivity was not significantly reduced by exposure to herbicides in S. feltiae but H. bacteriophora was less tolerant. Synergistic effect was obtained in the nematode mortality test but no synergistic effect was observed in the nematode infectivity assay. Our results suggest that possible synergistic effects of agrochemicals on survival of nematodes should be tested before mixing with entomopathogenic nematodes.     

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.     

Application methods of entomopathogenic nematodes for control of Mahanarva spectabilis (Hemiptera: Cercopidae)

The pathogenicity of entomopathogenic nematodes (EPNs) against nymphs of the pasture spittlebug Mahanarva spectabilis (Distant, 1909) (Hemiptera: Cercopidae) was studied under four application methods. Nymphs of the fourth or fifth instar were placed on the roots of elephant grass plants and submitted to four EPN isolates (Steinernema carpocapsae, S. feltiae, S. riobravis and Heterorhabditis amazonensis RSC1), at two concentrations (2000 and 4000 infective juveniles/mL), with four application methods (pipetting, spraying on the nymphs after froth formation, spraying before froth formation and placement of infected host cadaver method). There was no significant difference in the nymph mortality in function of the concentration and/or the isolate used. However, the efficacy was influenced by the application method, with the most efficient being sprayed on nymphs after froth formation and infected host cadavers. Steinernema riobravis applied by spraying on nymphs with froth, at a concentration of 2000 EPNs/mL, and H. amazonensis RSC1 applied by infected host cadavers caused 71% of the nymphs to die. The use of infected host cadavers and spraying in an aqueous solution on nymphs after froth formation were found to be the most efficient methods to control Mahanarva spectabilis.     

Survey of entomopathogenic nematodes and associate bacteria in Thailand and their potential to control Aedes aegypti

Aedes aegypti is an insect vector that transmits several viruses affecting humans worldwide. Entomopathogenic nematodes (EPNs) and their symbiotic bacteria are organisms with the potential to control many insects. In this study, we did a survey aimed to identify EPNs and their symbiotic bacteria and evaluate the larvicidal activity of bacteria against Ae. aegypti. We collected 540 soil samples from 108 sites in Phitsanulok Province, lower northern Thailand. Baiting techniques and White traps were used to isolate EPNs from soil samples. By sequencing of 28S rDNA and internal transcribed spacer regions, 51 EPN isolates were identified as Steinernema surkhetense (35 isolates), Heterorhabditis indica (14 isolates) and Heterorhabditis sp. SGmg3 (two isolates). Based on sequencing of a partial region of the recA gene, 35 isolates of Xenorhabdus were identified as Xenorhabdus stockiae, and 20 Photorhabdus isolates were identified as Photorhabdus luminescens subsp. akhurstii (10 isolates), P. luminescens subsp. hainanensis (seven isolates) and P. asymbiotica subsp. australis (three isolates). Screening for larvicidal activity of bacteria against Ae. aegypti was performed in the laboratory. Xenorhabdus WB5.4 and Xenorhabdus WB12.5, which were closely related to X. stockiae, resulted in high mortality of Ae. aegypti (99.99% and 70%, respectively) at 96 hr after exposure. Comparing with control groups, mortality of Ae. aegypti larvae was low (1.11%–6.67%) after exposure for 24–96 hr. Our findings showed the potential of X. stockiae for controlling Ae. aegypti. Further studies are needed to elucidate the mechanisms through which these bacteria kill Ae. aegypti larvae.     

Nutritive significance of crystalline inclusion proteins of Photorhabdus luminescens in Steinernema nematodes

Phase I cells of Photorhabdus luminescens produce two types of intracellular crystalline inclusion proteins designated CipA and CipB. The genes encoding CipA and CipB proteins from P. luminescens H06 were expressed respectively in Escherichia coli and these cells were used to feed the axenic first juveniles (J1) of three Steinernema nematode isolates in liquid cultures and on agar plates. In liquid cultures, the axenic J1 juveniles of all three test Steinernema nematode isolates were able to produce next dauer juveniles (DJs) in the E. coli cultures with at least one of the expressed Cip proteins, but unable to develop beyond the next J1 stage without expressed Cip proteins. For each target nematode isolate, addition of the supernatant of the bacterial culture of its Xenorhabdus symbiont to the tested liquid cultures did not induce the formation of DJs. However, on LB agar plates with different test E. coli cultures, all J1 juveniles of the three Steinernema strains finally developed into next DJs. It seemed that the metabolite pathway of the test bacteria in both culture systems was different. The presence of the Cip proteins has a significant influence on the DJ formation of the Steinernema nematodes in liquid culture system.     

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.     

Biological control potential of native entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) against Spodoptera cilium (Lepidoptera: Noctuidae) in turfgrass

In laboratory studies, we demonstrated that five native entomopathogenic nematode species/isolates caused 100% mortality of Spodoptera cilium larvae, a soil surface-feeding pest of turfgrass. At 25 infective juveniles/cm² applied to sod, two selected Turkish species, Steinernema carpocapsae and Heterorhabditis bacteriophora (Sarigerme isolate), averaged 77% and 29% larval mortality, respectively.     

Isolation and virulence of entomopathogenic fungi against larvae of hazelnut weevil Curculio nucum (Coleoptera,Curculionidae) and the effects of combining Metarhizium anisopliae with entomopathogenic nematodes in the laboratory

An approach to ensure effective pest biocontrol would be to select native isolates of biological control agents (BCAs). A survey to isolate entomopathogenic fungi (EPF) from a hazelnut growing area has been carried out. EPF were recovered from 133 of the 295 soil samples. The main species isolated were Metarhizium anisopliae sensu lato (36%) and Beauveria bassiana sensu lato (42.8%). With regard to controlling abiotic factors, altitude had an effect on the distribution of B. bassiana, but not on M. anisopliae. Cropping system did not have an effect on the occurrence of these EPF, while pH appeared as a predictive variable for both. In addition, we tested the virulence of six of these isolates: three M. anisopliae and three of B. bassiana against larvae of Curculio nucum L. The highest larval mortality (reaching 80%) was due to M. anisopliae (strain 34) when applied in simultaneous combination with four entomopathogenic nematode species: Steinernema carpocapsae (strain B14), Steinernema feltiae (strain D114), Steinernema. sp. (strain D122) and Heterorhabditis bacteriophora (strain DG46). The effect of nematodes was greater and no antagonistic or synergistic effects were observed.     

Pathogenicity of five species of entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) to the ixodid tick Dermacentor silvarum Olenev (Acari: Ixodidae)

We determined the mortality of adult tick Dermacentor silvarum Olenev after it was exposed to five entomopathogenic nematode species: Steinernema carpocapsae BJ (Sc BJ), S. glaseri NC32 (Sg NC32), S. ceratophorum D43 (Sc D43), S. feltiae Otio (Sf Otio) and Heterorhabditis bacteriophora E-6-7 (Hb E-6-7). All five species had lethal effects on adult ticks, and caused significant decreases in the amount of eggs laid by engorged female ticks (P < 0.05). Among them, Hb E-6-7 and Sc BJ were the most pathogenic to female adult ticks, with mortalities ranging from 70 to 90% (P < 0.05). Engorged females were more susceptible than unfed and partially fed females. The cumulative mortalities of engorged females exposed to Hb E-6-7 and Sc BJ for 14 days were 90%, with LT₅₀ at 6.3 and 7.0 days, respectively. Dose-dependent effect was observed in the oviposition of the ticks after they were exposed to nematodes at concentrations of 1000–5000 IJs/dish. When the concentration was higher than 5000 IJs/dish, dose dependent effects disappeared (P > 0.05). All five nematode species were lethal to unfed male ticks, and there was significant difference in mortality when compared to the control group (P < 0.05); however, the pathogenicity varied among them.     

Production technology for entomopathogenic nematodes and their bacterial symbionts

Entomopathogenic nematodes (genera Steinernema and Heterorhabditis) kill insects with the aid of mutualistic bacteria. The nematode–bacteria complex is mass produced for use as biopesticides using in vivo or in vitro methods, i.e., solid or liquid fermentation. In vivo production (culture in live insect hosts) is low technology, has low startup costs, and resulting nematode quality is high, yet cost efficiency is low. In vitro solid culture, i.e., growing the nematodes and bacteria on crumbled polyurethane foam, offers an intermediate level of technology and costs. In vivo production and solid culture may be improved through innovations in mechanization and streamlining. In vitro liquid culture is the most cost-efficient production method but requires the largest startup capital and nematode quality may be reduced. Liquid culture may be improved through progress in media development, nematode recovery, and bioreactor design. A variety of formulations is available to facilitate nematode storage and application. Journal of Industrial Microbiology & Biotechnology (2002) 28, 137–146 DOI: 10.1038/sj/jim/7000230 Received 16 August 2001/ Accepted in revised form 10 November 2001     

昆虫病原线虫对腰果细蛾的致病力测定

室内测定了Steinernema carpocapsae All、S.carpocapsae Biosys、S.glaseri NC52、S.longicaudum X-7、Heterorhabditis bacteriophora H06和H.indica LN2共6个昆虫病原线虫品系对腰果细蛾Acrocercops syngramma Meyrick幼虫的致病力。结果表明,线虫S.carpocapsae All、Biosys品系与线虫S.glaseri NC52对腰果细蛾幼虫的致死效果显著高于其它线虫,但供试的6个线虫品系均未能在腰果细蛾幼虫体内繁殖下一代,说明供试线虫在腰果细蛾幼虫体内难以繁殖。选择小卷蛾斯氏线虫S.carpocapsae All进一步研究其对腰果细蛾的致病力,结果发现,All线虫对腰果细蛾3龄幼虫处理24 h后的致死中浓度LC50为19.88 IJs/虫。腰果细蛾各龄幼虫和蛹对小卷蛾斯氏线虫S.carpocapsae All均表现出高度的敏感性,但不同幼虫龄期和处理时间敏感程度不同。小卷蛾斯氏线虫S.carpocapsae All对不同龄期腰果细蛾幼虫和蛹均表现出较强的致病力。