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.     

Comparative effects of Steinernema feltiae (Nematoda: Steinernematidae) and insecticides on yield and cropping of the mushroom Agaricus bisporus

In a mushroom crop (Agaricus bisporus) affected by a very low level of sciarid fly (Lycoriella auripila) infestation, the effects of an indigenous isolate of insect-parasitic nematode (Steinernema feltiae) and of two commonly used insecticides (diazinon and diflubenzuron) were studied. When compared with untreated plots, nematodes applied to the casing had no adverse effects on mushroom yields whereas insecticides decreased yields. At a rate of 3 × 10⁶ infective juveniles per tray (surface area = 0.56 m²), S. feltiae elicited increases of 28.5% and 19% in the mean total numbers and weights of mushrooms respectively. Treatment only with diflubenzuron resulted in 14.6% and 6% reductions in mean total numbers and weights of mushrooms, respectively; treatment with both diazinon and diflubenzuron caused 18.5% and 9.4% losses. Application of nematodes generally reduced the mean weight per mushroom whereas insecticides increased it; nematodes delayed the onset of mushroom production (first flush) whereas diflubenzuron delayed the third and fourth flushes. Nematode contamination of sporophores was minimal when S. feltiae was applied at casing. Although their numbers declined with time, the nematodes persisted, in the casing layer, throughout the cropping period of seven weeks. It is concluded that yield benefits associated with nematode application can result mainly from nematode effects on A. bisporus and not solely from suppression of a damaging pest population.     

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.     

Comparison of the Efficacy of Entomopathogenic Nematodes for the Biological Control of the Mushroom Pests Lycoriella auripila (Sciaridae) and Megaselia halterata (Phoridae)

The efficacy of different species of entomopathogenic nematodes was tested against larvae of the mushroom phorid Megaselia halterata (Diptera: Phoridae) and the mushroom sciarid Lycoriella auripila (Diptera: Sciaridae). Sciarid larvae originating from infestations in casing soil during colonization by Agaricus bisporus were almost completely controlled by applications of Steinernema feltiae to the casing soil. When larvae originated from infestations in freshly spawned compost, they could be controlled by compost applications halfway through spawnrunning and by very early casing treatments. The control of phorids in compost was maximally 31% when nematodes were mixed within the infested compost at a concentration of 3 106 nematodes/m2. Only slightly higher reduction rates were obtained at higher concentrations. The control of phorids was more promising in the infested casing layer, in which S. carpocapsae was most successful. At concentrations of 6 and 15 106 nematodes/m2 this species obtained reduction rates of 65 and 73% respectively when it was applied 3 days after the end of the infestation period. These concentrations are, however, too high for practical application.     

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.     

Biological control of sciarid and phorid pests of mushroom with predatory mites from the genus Hypoaspis(Acari: Hypoaspidae) and the entomopathogenic nematode Steinernema feltiae

In small-scale experiments, the predatory mites, Hypoaspis aculeifer (Canestrini) and H. miles Berlese, applied at 700 mites m(-2), and the entomopathogenic nematode, Steinernema feltiae (Filipjev) applied at 3 x 10(-6) nematodes m(-2) controlled sciarids and phorids in mushroom compost and casing substrates. For both mite species, earliest application to the growing substrate following sciarid infestation reduced sciarid emergence. In contrast, later application of each biological control agent provided more effective control of phorid emergence. The behaviour of adult mites suggested that H. aculeifer were more positively geotactic than H. miles although both species could penetrate compost and casing substrates to a depth of 2-12 cm. A majority of S. feltiae nematodes resided at a depth of 2-4 cm in both substrate types. Independent application of H. aculeifer provided more comprehensive control of sciarids and phorids than the other biological agents studied, owing to its better dispersal within compost and casing, and ability to attack larvae of differing ages.     

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.     

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.     

Effects of the paedogenetic mushroom cecid, Heteropeza pygmaea (Diptera:Cecidomyiidae) on cropping of the cultivated mushroom (Agaricus bisporus)

When introduced into a mushroom crop at rates of 2, 20 or 200 larvae/tray (0.56 m²), the mushroom cecid, Heteropeza pygmaea, caused significant reductions in both yield and number of mushrooms in relation to the infestation level. The reductions were greater when the larvae were introduced at spawning rather than at casing. The yield and number of infested (unmarketable) mushrooms increased significantly in relation to the initial infestation level. Just two H. pygmaea larvae, introduced at spawning, resulted in cecid populations that caused a 12% loss in total yield in addition to a 7% loss due to spoilage. Loss assessment in the future, therefore, should take into account both yield suppression and spoilage. There was little effect of cecid infestation on flush timing and mushroom size was only affected in the fourth flush, when a significant reduction (27%) was shown at the highest infestation rate at spawning.     

Selective breeding of Steinernema feltiae (Filipjev) (Nematoda: Steinernematidae) for improved efficacy in control of a mushroom fly,Lycoriella solani Winnertz (Diptera: Sciaridae)

A method of selecting a Steinernema feltiae strain that is effective against a mushroom fly, Lycoriella solani, is described in detail. The pest control efficacy of the selected nematode strain was evaluated and compared with the efficacy of two unselected strains. The selection procedure was designed to give preference to nematode individuals with the greatest ability (1) to search effectively for the target insect larvae in their natural habitat, (2) to infect them shortly after application and (3) to reproduce in their haemocoel. Thirty‐four rounds of selection achieved a 4‐fold improvement in nematode ability to find and parasitize third‐ and fourth‐instar larvae of the pest in the mushroom substrate. In 24‐h laboratory experiments, mortality of the insect caused by nematode juveniles rose from 22.5%, recorded for the original unselected isolate, to 92.5% for the selected strain. In a 51‐day experiment conducted on a mixed age mushroom house population of L. solani, the enhanced pest control ability of the selected strain was detected shortly after nematode application and remained high throughout the experimental period. During the first 4 weeks of the trial the selected nematode strain was significantly better than both unselected strains and caused 91.1–92.7% reduction of the fly emergence from the mushroom substrate. No difference was observed between the efficacy of the selected nematodes applied at 1 × 10⁶ and 3 ×10⁶ infective juveniles per m², while the unselected strains performed significantly better at the higher concentration. All the nematodes examined showed good persistence in the mushroom casing apparently due to recycling in the insect host.     

Susceptibility of larvae of the mushroom fly Megaselia halterata to the entomopathogenic nematode Steinernema feltiae in bioassays

Bioassays were initially conducted in Petri-dishes to screen the efficacy of four Heterorhabiditis and Steinernema species against the mushroom phorid Megaselia halterata. Control rates of 61 to 70% control were obtained at a dosage of 1500 infective juveniles (IJs) per 30 larvae. In order to avoid stress-induced susceptibility, an improved bioassay system in micro-wells, filled with 0.6 ml of compost agar and 0.2 of compost colonized by Agaricus bisporus, was developed. In a screening of different species of Heterorhabiditis and Steinernema with applications of 30 IJs per phorid larva, a highest parasitization rate of 20% was obtained with S. feltiae. Bioassays were continued with S. feltiae in dosage-mortality assays in which larvae of the sciarid Lycoriella auripila and the phorid M. halterata were challenged. At the lowest dosage of 30 IJs per sciarid larva, 78% control was obtained. Increasing the dosage from 30 to 1000 led to only small increases of the phorid mortality. At 1000 IJs per larva a significant mortality of 18% was obtained. The nature of the substrate, compost or casing did not greatly influence the parasitization rates. The sex ratio of nematodes that were able to penetrate and establish in the phorid larvae appeared to be female-skewed. Males were only present at a mean of 19%. Low susceptibility of the phorid larva was ascribed to the inaccessibility of its small mouth opening.     

Effects of Entomopathogenic Nematodes on Control of a Mushroom-infesting Sciarid Fly and on Mushroom Production

The entomopathogenic nematodes Steinernema feltiae (Biosys strain #27) and Heterorhabditis heliothidis were evaluated for the larval control of a mushroom-infesting sciarid, Lycoriella mali, and for the effects of these nematodes on mushroom (Agaricus bisporus) production. In a series of small-scale mushroom crops, infective-stage H. heliothidis and S. feltiae were applied to the mushroom casing surface in the irrigation water or incorporated into the casing material at densities ranging from 28 to 1120 and 11 to 1120 nematodes cm-2 of casing surface respectively. The mortality of L. mali larvae ranged from 52 to 100% for H. heliothidis and 38 to 100% for S. feltiae. Both nematode species reduced mycelial coverage on the casing surface at primordia initiation. Neither mushroom strain (off-white or white hybrid) or method of application (incorporation into or irrigation onto the casing surface) altered the effect on mycelial coverage. The nematodes's negative effect on mycelial growth confounded the benefit of fly control. At high nematode densities (up to 1120 nematodes cm-2), damage-free mushroom yields for the first week of harvest were less than those from the untreated control. However, at lower nematode densities, at or below 140 cm-2, the nematodes had less effect on mushroom growth, and consequently, damage-free mushroom yields for the first week of harvest were frequently greater than those from the untreated control. In the absence of flies, the first-week mushroom yield generally declined with increasing nematode densities for both white and off-white mushroom hybrids. After 4 weeks of harvest, accumulated mushroom yields had nearly recovered from the earlier decline.     

Physiological studies of a hybrid between populations of Dactylis glomerata from contrasting climatic regions. II. Intra population variation

Mushroom compost was treated with nematicides and infested with Aphelenchoides composticola at the time of filling into growing containers. Yields of mushrooms from infested untreated control composts were reduced to 40–60% of yields from uninfested control compost. Yields from infested compost treated with fenamiphos emulsifiable concentrate (e.c.) at 10 or 20 mg a.i./kg, thiabendazole wettable powder at 40 or 60 mg a.i./kg or oxamyl granules at 20 mg a.i./kg were as high as from uninfested controls. Compost treated with granules of AC 64,475 up to 20 mg a.i./kg or ethoprophos or thionazin up to 80 mg a.i./kg gave yields significantly lower than uninfested controls. Numbers of nematodes rose to about 10⁶/20 g of compost in untreated compost and then fell, and a similar peak occurred in treatments in which yields were substantially reduced by nematode damage. Treatments which yielded as well as the uninfested controls held maximum nematode numbers down to about 10V20 g of compost but populations stayed at this level or tended to rise while numbers in untreated compost fell. Incorporation of fenamiphos in casing or its application to the surface of beds 3 wk after cropping began gave lower yields than the uninfested control but mushrooms were being produced late in the cropping cycle. Fenamiphos e.c. at 20 mg ai./kg incorporated in compost is considered a practical preventive measure for control of A. composticola.     

Effects of entomopathogenic nematodes on the spruce web‐spinning sawfly Cephalcia arvensis panzer and its parasitoids in the field

Field tests were conducted in the Asiago Forest, Venetian Prealps (Italy) to evaluate the efficacy of nematode strains (Heterorhabditis sp. HL 81 Steinernema carpocapsae IS 230 S. feltiae C = bibionis,) IS 389 S. kraussei SK) against the spruce web‐spinning sawfly Cephalcia arvensis. Soil applications of 100 juveniles cm^‐2 of S. feltiae and S. kraussei resulted in 56% and 36.4% reduction of emergences of sawfly, respectively, when performed before the mature larvae drop and enter the soil. The effectiveness of S. feltiae becomes 32.3% if the nematodes are applied when the larvae have already prepared their chambers. S. feltiae parasitized more females and long‐term diapausing individuals than S. kraussei. The two most effective strains (IS 389 and SK) seem to be well adapted to low temperature, which is likely to be the most important limiting factor for nematode activity in the mountain spruce forests. An ichneumonid parasitoid (Xenoschesis fulvipes,) was strongly affected by S. feltiae, resulting in 66.6% reduction of emergences. Another ichneumonid Ctenopelma lucifer, seems to be less affected than X. fulvipes by the nematode application.     

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.     

The control of paedogenetic cecid larvae in mushroom beds by the use of thionazin

Thionazin incorporated in the casing layer of mushroom beds was shown to be highly phytotoxic even at 5 ppm, but when incorporated in compost it caused little phytotoxicity up to 20 ppm. A rate of 8 ppm gave satisfactory control of Heteropeza pygmaea Winnertz (Dipt.: Cecidomyiidae), but was less suitable against Mycophila speyeri (Barnes), for which the use of γ-BHC in the casing should be retained. A severe infestation of M. speyeri was shown to depress the total yield of mushrooms by 59% and the saleable yield by 87%.     

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.     

Effects of bendiocarb and diflubenzuron on mushroom cropping

When mixed into the casing or compost layers of a mushroom bed in the absence of pests, bendiocarb decreased yield and number of mushrooms according to concentration. The most severe effects were on mushroom number at the two highest rates used (100 and 1000 μg/g), and there were large increases in mushroom size. Effects of bendiocarb incorporation in the compost diminished with time, and there was partial compensation in yield and numbers at the fourth flush. The action of bendiocarb persisted when it was mixed into the casing. Diflubenzuron showed some opposite effects at lower concentrations. When either mixed into, or drenched onto the casing at the commercial rate (30 μg/g), yield and size were both increased and the timing of the flushes was unaffected. At the two higher concentrations (180 and 1080 μg/g), reductions in yield and number and an increase in mushroom size were shown. However, these effects became more severe with time, especially those on mushroom number, possibly due to the accumulation of a toxic breakdown product.     

Influences of compost and casing layer depths on the mechanical properties of mushrooms

The mushroom growing bed consists of an underlying compost which is covered with casing (a peat and chalk or lime mixture) to induce fruit-body formation. The influences of the depths of the compost and casing layers on the mushroom texture, susceptibility to bruising, and ease of detachment from the growing bed surface were investigated. Textural properties were determined on cubes of cap tissue and detachment torque was determined by using an instrumented rotating suction cup which was lowered on to the caps. The proportion of mushrooms which detached at the base from the bed surface was recorded, since this is an important parameter for an automatic (robotic) harvesting system. Shallow casing resulted in mushrooms that were firmer and less likely to deform plastically, but that showed greater discolouration after a standard bruising treatment. Shallower casing also increased the proportion of mushrooms that detached at the base. Increasing the depth of compost produced firmer mushrooms, significantly increased the detachment torque and reduced the proportion of mushrooms detached at the base. The proportion of mushrooms that detached at the base correlated negatively with the plastic deformation of cap tissue. A positive correlation was found between mushroom dry matter content and tissue firmness.     

The effect of some microphagous saprobic nematodes on mushroom culture

When added to spawned mushroom compost the microphagous saprobic nematodes Pelodera cylindrica, Mesorhabditis spiculigera and Acrobeloides bütschlii did not affect mycelial growth or compost pH. The eelworms multiplied at first but their numbers declined as the mycelium colonized the compost. In contrast, the fungal-feeding Aphelenchoides composticola increased rapidly and destroyed the mycelium. Excess water, and the presence of the fungal competitor of mushroom, Chaetomium olivaceum, allowed some increase of the microphagous forms. Different numbers of Mesorhabditis spiculigera added to the ‘casing’ had no effect on mushroom yield but the mushroom ‘flushes’ seemed less pronounced. Reasons for the failure of the saprobic eelworms to affect mushroom, and the possibility of synergistic pathogenicity by these eelworms and bacteria, are discussed.