Comparative Assessment of biological (Nematoda: Steinernema feltiae) and chemical methods of control for the mushroom fly Lycoriella auripila (Diptera: Sciaridae)

The sciarid fly Lycoriella auripila is the major pest of mushrooms cultivated in the UK. Its larvae, which are capable of damaging the crop at all stages of production, may cause severe yield losses and can only be controlled with chemical pesticides. An indigenous isolate of the insect‐parasitic nematode Steinemema feltiae was tested as a biological control agent and its effects compared with two commonly used insecticides, diazinon and diflubenzuron. The timing of application of nematodes was found to affect their efficacy. When applied to compost during spawning, nematodes did not significantly reduce fly emergence, but they did if applied at casing when they were almost as effective as diflubenzuron. Diazinon incorporated into compost did not reduce fly emergence and was also the only treatment that did not lower the incidence of mushrooms spoiled by tunnelling of the larvae of L. auripila. When compared with untreated control plots those treated both with diazinon and diflubenzuron showed significant mean losses in yield of 10% in total weight and 17% in total numbers of mushrooms picked. In contrast, when S. feltiae was applied at casing significant mean increases in yield of 7% and 19%, respectively, were attained. Infective nematodes persisted well in casing, very few were found on sporophores.     

Effects of Caenorhabditis elegans (Nematoda: Rhabditidae) on yield and quality of the cultivated mushroom Agaricus bisporus

Thirteen species of saprobic rhabditid nematodes (11 genera) were identified from samples of compost and casing material collected from mushroom farms in the British Isles. Caenorhabditis elegans, the most frequently found saprobe, was mass-produced monoxenically and its effects on the cultivated mushroom, Agaricus bisporus (strain U3) were studied. C. elegans did not multiply in well-prepared, pasteurised, spawned compost, whereas casing material proved to be a highly suitable environment for its reproduction. An initial casing inoculum of 10⁶ nematodes/crate of compost (7.5 kg), caused a significant reduction in mushroom yield. Losses in total mushroom yields of 11%, 20% and 26% were caused by initial inoculum rates of 10⁶, 10⁷and 2 × 10⁷ nematodes/crate, respectively. Yields were negatively correlated with the initial nematode inoculation level and regression equations were derived. The nematode treatments caused fewer mushrooms to be produced and an absence of the usual distinctive flushing patterns. C. elegans caused considerable deterioration in mushroom quality and characteristic distortion of mushrooms. Individual sporophores were mis-shapen, notched and had brown or violet coloured grills. Up to 3.8%, 6.7% and 10.8% of total weight and 3.5%, 5.4% and 8% of total numbers of mushrooms were distorted at the three highest nematode inoculum rates tested. Weights and numbers of distorted mushrooms were positively correlated with the initial nematode population. C. elegans commonly colonised sporophores.     

Influence of Steinernema feltiae and Diflubenzuron on Yield and Economics of the Cultivated Mushroom Agaricus bisporus in Dutch Mushroom Culture

The insect pathogenic nematode Steinernema feltiae (Nematoda: Steinernematidae) was shown to offer an alternative to the use of diflubenzuron for the control of the mushroom fly Lycoriella auripila (Diptera: Sciaridae). The influence of diflubenzuron and S. feltiae treatments on the yield and numbers of the mushroom, Agaricus bisporus , was studied. Applications of diflubenzuron after casing significantly reduced total mushroom yields by 10-13%. The greatest yield reductions were observed in the first flush. In the second flush, high yields were found despite significantly lower numbers of mushrooms. These high yields, however, could not compensate for the yield loss of the first flush. Some of the treatments with S. feltiae significantly increased yields, but these were not associated with a particular time of application. As nematodes did not reduce mushroom yields they were able to compete with diflubenzuron. Although the purchase costs of nematodes were high, their use was more economical than that of diflubenzuron when the yield losses of 10% due to casing treatments were taken into account.     

Comparative effects of three insect growth regulator insecticides and a dipteran-active strain of Bacillus thuringiensis on cropping of the cultivated mushroom Agaricus bisporus

Three insect growth regulator insecticides and an entomopathogenic strain of Bacillus thuringiensis (GC327), products effective against the mushroom sciarid, Lycoriella auripila, were compared for their effect on mushroom cropping. Cyromazine and diflubenzuron were applied as a surface drench to mushroom compost before or after pasteurisation (at filling or spawning, respectively); admixed into casing material (at casing); or at a combination of these times. Hexaflumuron and GC327 were applied only at filling and casing, respectively. The presence of the target pest, L. auripila, had no effect on treatment trends, although it was accounted for in the analysis by use of a yield model. The trial was notable for the disparate effects that cyromazine and diflubenzuron casing treatments had on mushroom cropping. Cyromazine treatments that included application at casing resulted in increases in yield, compared to the untreated control whereas, with diflubenzuron, the opposite was true, with treatment at casing alone causing the greatest reduction overall (10%). GC327 applied at casing was also conspicuous for giving a 13% increase in yield. Treating the crop at casing with either cyromazine or GC327, therefore, resulted in a 15% or 24% increase in yield, respectively, compared to a similar treatment with diflubenzuron. Hexaflumuron applied at filling caused increases in yield compared to application of cyromazine at filling and cyromazine or diflubenzuron at spawning. There were also effects on crop timing. The addition of a cyromazine casing treatment normally caused the distinct flushes of mushrooms to be produced significantly earlier than the untreated control (up to 2.5 days), as did GC327. With diflubenzuron, the earlier flushes were only produced by those treatments that did not include a casing application. The combinations that included a casing treatment with diflubenzuron initially produced mushroom flushes earlier than the untreated control. They became either synchronous with the control or they were delayed. From the crop tolerance perspective, therefore, cyromazine and GC327 would be the sciarid control products of choice for a commercial mushroom grower.     

Effects of Application Rates of Steinernema feltiae (Nematoda: Steinernematidae) on Biological Control of the Mushroom Fly Lycoriella auripila (Diptera: Sciaridae)

The potential of Steinernema feltiae for the biological control of Lycoriella auripila was tested in commercial mushroom‐growing conditions. The nematodes, applied at rates of 1.5, 3, 6 or 12 x 10 ⁶ infective juveniles per 34 kg tray of spawn‐run compost, were mixed into the casing material before it was spread over the compost surface. When compared with untreated control trays, any rate of nematode application significantly reduced fly emergence. Insecticides significantly reduced mushroom yields; nematodes significantly increased them. At a rate of 3 x 10 ⁶ infectives/tray S. feltiae elicited mean total increases in the weight and numbers of mushrooms produced of 8% and 11% respectively. The nematodes also reduced the incidence of mushrooms spoiled by tunnelling sciarid larvae. The early decline in the numbers of nematodes persisting in casing was a trend that was reversed later, when evidence was obtained that S. feltiae was recycling in insects that had been killed. When applied at a rate of 3 ‐10⁶infectives/tray of compost S. feltiae should provide reliable and cost‐effective biological control of L. auripila.     

Factors affecting entomophilic activity of Neoaplectana feltiae in mushroom compost

The host-searching ability of Neoaplectana feltiae Filipjev (= S. bibionis Bovien) (Nematoda: Steinernematidae) in response to larvae of a mushroom fly, Lycoriella solani Winn. was examined in a mushroom substrate. Individuals of L. solani were less attractive for the parasite than larvae of Galleria mellonella L. The nematode juveniles penetrated a 22 cm layer of casing mixture within 2–4 days. In the casing alone nematode effectiveness was better than in mushroom compost or in compost and casing together. In the casing mixture parasite dosages of 20 and 100 juveniles per cm² led to 22% and 45% parasitization of L. solani respectively, while all G. mellonella larvae were parasitized at both dosages. The prevalence of nematode infection depended on the content of water in the mushroom substrate. The highest N. feltiae infectivity was observed, when the ratio of the dry casing weight to the weight of water content was 1: 2.5. The practical aspects of the observed phenomena, essential for the use of N. feltiae in the protection of commercial mushroom cultivation are discussed.     

Control of the mushroom pests Lycoriella auripila (Diptera: Sciaridae) and Megaselia halterata (Diptera: Phoridae) by Steinernema feltiae (Nematoda: Steinernematidae) in field experiments

The nematode Steinernema feltiae (Nematoda: Steinemematidae) was tested for its ability to control two main mushroom pests i.e. the sciarid Lycoriella auripila (Diptera: Sciaridae) and the phorid Megaselia halterata (Diptera: Phoridae) in growing-rooms filled with spawned compost. A clear difference between female and male sciarid control was observed. A nematode application 1 day after casing preceded by an application 1 day before casing on the compost caused an almost complete control (97%) of the F1-generation of female sciarids. The F2-generation of females was similarly controlled (95%) by an application 7 days after casing. A dosage of 1 × 10⁶nematodes m^-2was found to be equally effective as higher dosages. Diflubenzuron remained active throughout entire the cropping period with high sciarid mortality rates varying from 72% to 99%. Phorid control was variable and seemed to depend on the presence of sciarids. In one occasion the control rate of F2-generation phorid larvae was 75% and was possibly caused by the presence of new infective juvenile nematodes recycled in F2-generation sciarid larvae. Diflubenzuron did not significantly reduce phorid numbers.     

Evaluation of a genetically selected strain of Steinernema feltiae against the mushroom sciarid Lycoriella mali

The control potential of a strain (ScP) of Steinernema feltiae genetically selected against the mushroom sciarid Lycoriella solani, was evaluated for the management of L. mali. Trials were conducted at two commercial mushroom farms with high and low levels of fly infestation. The efficacy of the ScP strain was compared with the SN strain of S. feltiae and the chitin synthesis inhibitor, diflubenzuron. At low densities of L. mali, the two strains did not differ in efficacy and caused 85–94% reduction in fly populations. At high fly densities with a mixed infestation of the phorid fly, Megaselia halterata, the ScP stain caused 56–83% reduction in L. mali populations whereas the SN strain caused 51–73% reduction. Two doses of the improved strain (0.5 times 10⁶ and 1.0 times 10⁶ infective juveniles per m² cropping area) did not differ in efficacy from diflubenzuron which caused 80% reduction in L. mali populations. The lower dose (0.5 times 10⁶) of the SN strain was less effective than diflubenzuron. The ScP strain had a major advantage over the SN strain in persistence in casing material. Nematodes had no significant effect on mushroom yields.     

Bendiocarb and diflubenzuron as substitute insecticides for endosulfan in commercial mushroom growing

Diflubenzuron and bendiocarb treatments of compost and casing soil at a rate of 1 g a.i./m² each, resulted in comparable or even improved compost and casing soil colonisation over endosulfan treatments (at a rate of 1.5 g a.i./m²). Alternating diflubenzuron/bendiocarb treatments were preferable to the reverse treatments, which tended to diminish total yield of mushrooms significantly by 1.4–3.9 kg/m². This effect was due to the casing soil treatment with diflubenzuron and not due to the compost treatment with bendiocarb after spawning. The effectiveness of these substitutes for endosulfan was tested on five commercial mushroom farms, where a similar or improved control of Megaselia halterata (Diptera: Phoridae) was obtained over control (endosulfan) treatments. If present, Lycoriella auripila (Diptera: Lycoriidae) was very effectively controlled by the substitute insecticides compared with endosulfan. Relatively high numbers of M. halterata were occasionally observed in endosulfan-treated houses, indicating that a certain level of resistance towards this insecticide may already have developed. This assessment of tolerance in M. halterata emphasised the need for substitute insecticides with different modes of action, in addition to environmental reasons. The chemicals should be alternately applied within individual crops to avoid resistance development.     

Relative contribution of nematodes (Caenorhabditis elegans) and bacteria towards the disruption of flushing patterns and losses in yield and quality of mushrooms (Agaricus bisporus)

Laboratory tests of bacteria isolated from the body surface, or from the gut, of a saprophagous rhabditid nematode Caenorhabditis elegans infesting mushrooms (Agaricus bisporus) showed that some bacteria enhanced nematode reproduction and that others inhibited it. As some bacteria were shown to inhibit mycelial growth of the mushroom, the effects of Acinetobacter calcoaceticus var. anitratus, Enterobacter cloacae and Serratia liquefaciens, either alone or in combination with C. elegans, on the flushing patterns, quality and yield of A. bisporus (strain Horst U3) were studied. Bacteria alone had little effect on flushing patterns whereas C. elegans delayed the onset of mushroom production and significantly disrupted the growth pattern of crops, with mushrooms appearing more regularly and not within obvious flushes. Inoculation with bacteria resulted in ‘browning’ of mushrooms that was even more pronounced in C. elegans treatments. Characteristic distortion of sporophores was observed only in the presence of C. elegans. Nematodes commonly colonised sporophores. Bacteria affected the size of nematode populations both on the sporophores and in the casing. Significant yield loss occurred; up to 10% when bacteria were inoculated, up to 27.8% when C. elegans was inoculated, and up to 35% with both bacteria and nematodes. Synergism between C. elegans and A. calcoaceticus var. anitratus was observed; the combination resulted in significantly greater reduction in mushroom yield than any other treatment. It is concluded that bacteria contribute to yield loss and quality deterioration in A. bisporus but that the effects are far greater in the presence of C. elegans.     

Principles of insecticide action on mushroom cropping: incorporation into casing

The incorporation of organophosphorous insecticides into the casing layer of mushroom beds reduces the numbers of mushrooms, according to a logistic relationship with log₁₀ (dose). The cumulative numbers at any flush are affected in this way, and the response of individual flushes may, therefore, be represented by the difference between two logistic curves. This explains the observed increases in numbers in the second and third flushes following diazinon treatment and in the fourth following thionazin, pirimiphos-methyl and pirimiphos-ethyl. No compensation in numbers followed chlorfenvinphos treatment. This decreasing response pattern reflected the increasing residual action of the insecticides. Mushroom size was inversely related to numbers but even when numbers in any flush were unaffected by the treatment, there was usually an increase in size. Thus yield increases occurred in most flushes at commercial concentrations of four insecticides (thionazin compensated only in the fourth flush) and total yields were increased by 8 to 13%.     

Susceptibility of mushroom pests to the insect-parasitic nematodes Steinernema feltiae and Heterorhabditis heliothidis

The potential of two species of insect-parasitic rhabditid nematodes (Steinernema feltiae, Heterorhabditis heliothidis) for biological control of mushroom flies was studied in pot trials. Three Diptera that commonly infest mushroom crops were used; the larvae of Megaselia halterata (Phoridae), Heteropeza pygmaea (Cecidomyiidae) and Lycoriella auripila (Sciaridae) were all susceptible to parasitism by both nematode species. Fewer adult phorids and sciarids emerged when compost was nematode-treated and, for L. auripila, the effects of nematode applications at spawning, casing or on both occasions were compared. Casing treatments were more effective than spawning treatments; little extra benefit was gained from applying the nematodes twice. Populations of paedogenetic larvae of H. pygmaea built up rapidly in untreated compost, but were reduced when S. feltiae was applied, and were eradicated by H. heliothidis. Because they can penetrate insect cuticle, as well as natural body openings, Heterorhabditis spp. may be more suitable than Steinernema spp. for the control of mushroom fly larvae.     

Biology and control of the sciarid Lycoriella auripila Winn. (Diptera: Lycoriidae) in mushroom culture

The life-history of L. auripila, as a pest of mushrooms, is outlined and an explanation is given of the different types of damage caused. The most satisfactory control is based on the incorporation of insecticides in compost at spawning. Emulsions of chlorfenvinphos and diazinon at 50 ppm are lethal to young larvae for at least 7 weeks but granular formulations of the former must be applied at 100 ppm to achieve comparable results. Drenches of 0–01% malathion, applied at about 40 gal/1000 ft², are effective against larvae in the casing.     

Efficacy of soil treatments of entomopathogenic nematodes against the larvae, pupae and adults of Tuta absoluta and their interaction with the insecticides used against this insect

The efficacy of soil treatments of three native entomopathogenic nematodes (Steinernema carpocapsae, S. feltiae and Heterorhabditis bacteriophora) against Tuta absoluta larvae, pupae and adults was determined under laboratory conditions. The effect of three insecticides commonly used against T. absoluta, in the survival, infectivity and reproduction of these nematode strains was also evaluated. When dropped into soil to pupate, soil application of nematodes resulted in a high mortality of larvae: 100, 52.3 and 96.7 % efficacy for S. carpocapsae, S. feltiae and H. bacteriophora respectively. No mortality of pupae was observed and mortality of adults emerging from soil was 79.1 % for S. carpocapsae and 0.5 % for S. feltiae. The insecticides tested had a negligible effect on nematode survival, infectivity and reproduction. No sublethal effects were observed. Infective juveniles that survived to insecticide exposition were able to infect Galleria larvae with no significant differences from the control. The Galleria larvae affected by the three insecticides tested served as suitable hosts for the infection and reproduction of the nematodes. These results suggest that larvae of T. absoluta, falling from leaves following insecticide application, could be suitable hosts for nematodes, thereby increasing their concentration and persistence in the soil.     

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.     

Compatibility of Soil Amendments with Entomopathogenic Nematodes

The impact of inorganic and organic fertilizers on the infectivity, reproduction, and population dynamics of entomopathogenic nematodes was investigated. Prolonged (10- to 20-day) laboratory exposure to high inorganic fertilizer concentrations inhibited nematode infectivity and reproduction, whereas short (1-day) exposures increased infectivity. Heterorhabditis bacteriophora was more sensitive to adverse effects than were two species of Steinernema. In field studies, organic manure resulted in increased densities of a native population of Steinernema feltiae, whereas NPK fertilizer suppressed nematode densities regardless of manure applications. Inorganic fertilizers are likely to be compatible with nematodes in tank mixes and should not reduce the effectiveness of nematodes used for short-term control as biological insecticides, but may interfere with attempts to use nematodes as inoculative agents for long-term control. Organic manure used as fertilizer may encourage nematode establishment and recycling.     

The compatibility of the entomopathogenic nematode,Steinernema feltiae, and chemical insecticidesfor the control of the South American leafminer, Liriomyza huidobrensis

The compatibility of infectivejuveniles (IJs) of entomopathogenic nematodes, Steinernema feltiae, and chemical insecticides tocontrol larval stages of the South American leafminer,Liriomyza huidobrensis, was investigated.Initially the effect of direct IJ exposure to 5insecticides (abamectin, deltamethrin, dimethoate,heptenophos and trichlorfon) for 24 hours was testedagainst Galleria mellonella in a standard sandtube bioassay. Trichlorfon and dimethoate did notreduce the nematodes ability to locate and infect G. mellonella larvae to an unacceptable level. However,nematode infectivity was significantly reducedfollowing exposure to abamectin, deltamethrin andheptenophos. Secondly, IJ infectivity for L.huidobrensis in the presence of dry pesticideresidues on foliage was tested. No significantdetrimental effects on the level of control of L. huidobrensis was recorded when compared with theeffect of nematodes applied to residue free foliage.The integration of these agents into a pest managementprogramme is discussed.     

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.     

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.     

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.