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.     

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.     

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.     

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.     

Location of immature stages of the mushroom insect pest Megaselia halterata in mushroom-growing medium

Experiments were conducted to examine the location of oviposition by the phorid fly Megaselia halterata (Wood) (Diptera: Phoridae) in uncased and cased compost. Clearly, a majority of the gravid females choose oviposition sites directly after entering the top layer of the compost. In uncased compost, 60% of all adults emerged from the top of four compost layers of equal thickness. When the compost was covered by a casing layer which was still uncolonized by Agaricus bisporus, oviposition was further concentrated in the top compost layer. In this situation, 91% of all adults emerged from the top compost layer whereas only 1.5% emerged from the casing. When the casing layer was colonized by mushroom mycelium, 45% of all adults emerged from the casing layer and 53% emerged from the top compost layer. Further concentration in the top compost layer and the casing layer occurred as a result of upward migration of larvae. When compost was cased after oviposition, up to 43% of all adults emerged from the casing layer. We concluded that in the control of phorid infestations with insect pathogenic nematodes, applications in uncased compost can be restricted to the upper compost layer. When compost and casing are filled simultaneously, nematode applications in the casing layer only could be considered.     

Prey preference and life tables of the predatory mite Parasitus bituberosus (Acari: Parasitidae) when offered various prey combinations

Parasitus bituberosus Karg (Acari: Parasitidae) is one of the predatory mite species inhabiting mushroom houses. It is known to accept a wide range of prey, suggesting that it may be a promising candidate for the biological control of key pests of mushroom culture. In our study it did not show any prey preference among four groups of small organisms often occurring in mushroom growth medium, namely rhabditid nematodes, pygmephorid mites, and sciarid and phorid fly larvae. Nevertheless, the type of food these predators fed on affects their development. The shortest egg-to-adult development time was obtained on a nematode diet. On a diet of phorid larvae, mite development stopped at the deutonymph stage; none reached adulthood. All other diets sufficed to reach the adult phase. Female fecundity when fed nematodes and sciarid larvae did not differ, but it was much lower when fed pygmephorid mites. Other life table parameters confirmed that pygmephorid mites constituted the worst diet for P. bituberosus. The highest intrinsic rate of population increase (r _m  = 0.34) was obtained on the nematode diet; when fed sciarid larvae and pygmephorid mites it was 0.25 and 0.14, respectively. Our study provides good reasons to further test P. bituberosus as biocontrol agent of especially sciarid flies and nematodes, especially when the compost is well colonized by mushroom mycelium (which retards nematode growth).     

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.     

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.     

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.     

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.     

Transmission of the mycoparasite Coniothyrium minitans by collembolan Folsomia Candida (Collembola: Entomobryidae) and glasshouse sciarid Bradysia sp. (Diptera: Sciaridae)

Folsomia Candida was maintained on potato dextrose agar (PDA) plates precolonised by the mycoparasite Coniothyrium minitans for 3 yr but the sciarid Bradysia sp. survived for a maximum of only three generations. Collembolans and sciarid larvae from these cultures were able to transmit C. minitans to uninoculated PDA plates through the survival of spores in faecal pellets. Adult and larval sciarids also transmitted C. minitans from PDA culture to uninoculated PDA plates by contamination on the cuticle. In soil and sand both sciarids and collembolans were able to transmit C. minitans from C. m/m'tans-inoculated to uninoculated sclerotia of Sclerotinia sclerotiorum. Inoculation of sclerotia with C. minitans enabled greater populations of larger collembolans to develop. In the glasshouse where C. minitans had been applied to the soil, one adult sciarid and four collembolans out of 70 and 101 insects collected respectively yielded C. minitans on placement onto PDA + Aureomycin.     

Control of Western flower thrips (Frankliniella occidentalis Pergande) pupae in compost

Western flower thrips (Frankliniella occidentalis) develops from an active larval stage through to a non-feeding, almost immobile, pre-pupal and pupal stage. This generally occurs in the compost or soil below the plant on which the larvae fed. Control of thrips at this stage in their development offers a chance of utilising pathogens or pesticides not suitable for use in an integrated control programme aimed at adult and larval stages. Trials were done with F. occidentalis pupae and pre-pupae in a soil/peat based compost using 11 pesticides, three fungal pathogens and four species of insect parasitic nematodes. The pesticides malathion and chlorpyrifos-methyl gave the most promising result with 97.5% and 96.5% control, respectively. The fungal pathogen Metarhizium anisopliae proved better when applied as a pre-pupation rather than as a post-pupation treatment (74.5% : 26.6% control). The insect parasitic nematode Steinernema carpocapsae gave 76.6% control when applied at 25 times 10⁴ nematodes litre^-1 of compost. The results are discussed in relation to control of thrips in glasshouses.     

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.     

Instar Susceptibility of Simulium vittatum (Diptera: Simuliidae) to the Entomogenous Nematode Neoaplectana carpocapsae

Laboratory bioassays showed that the susceptibility of Simulium vittatum to Neoaplectana carpocapsae increased with successive larval instars. First, second, and third instar larvae were resistant to infection, while seventh instars were highly susceptible. Significant differences in intra-instar susceptibility were also evident, as mortality ranged from 58% for the smallest seventh instar larvae to 97% for the largest. Dissections revealed that the basis for the resistance of early instars was physical exclusion of the comparatively large nematodes. The principle factor regulating the susceptibility of mid and late instars was injury to nematodes caused by larval mouthparts during ingestion. Differences in intra-instar susceptibility were similarly related to nematode injury.     

Phoretic relationship between the myceliophagous mite Microdispus lambi (Acari: Microdispidae) and mushroom flies in Spanish crops

We studied the role played by the phorid Megaselia halterata (Wood) and the sciarid Lycoriella auripila (Winnertz) in the phoretic dispersion of the myceliophagous mite Microdispus lambi (Acari: Pygmephoridae). Twenty‐four crops were monitored during 18 months in commercial mushroom farms in Castilla‐La Mancha (Spain). Adults of both species were collected weekly and the mites they carried were counted and identified. Both phorids (19.6%) and sciarids (4.4%) carried the mite M. lambi. The calculated load of each was 3.4 M. lambi mites per phorid and 1.9 per sciarid. The same percentage of male and female phorid was used as vector, but the load was slightly higher for females (1.86 mites per female compared with1.48 mites per male). A mean of 7.2% of the phorids examined in winter were vectors of M. lambi, while in spring and autumn of the first year the average was more than 22%. The mean load did not vary significantly between seasons. Inside the mushroom farms, less than 10% of a small initial population of phorids carried mites (less than two mites per phorid). As the cycle progressed, more than 35% of a larger population of emerging flies did so (average 3.5 mites per phorid vector). At the end of the growth cycle, the flies may fly off to colonise nearby farms, favouring the propagation of M. lambi from infested to uninfested crops. Megaselia halterata is the principal vector of M. lambi in the mushroom farms of Castilla‐La Mancha due to their high numbers, the high percentage carrying mites and the number of M. lambi they carry.     

Inter- and intraspecific variation in infectivity ofSteinernema spp. to larvae of the mushroom flyLycoriella solani

Laboratory bioassays were done to assess the susceptibility of larvae of the mushroom sciarid flyLycoriella solani Winnertz (Diptera; Sciaridae) to 16 isolates comprising five species of the entomopathogenic nematode,Steinernema (Nematoda; Steinernematidae). Each isolate was tested by challenging sciarid larvae with single nematodes. Six isolates were also tested in dose-response experiments. Variations in nematode infectivity occurred (p<0.001) at both inter- and intraspecific levels.Steinernema feltiae (Filipjev) was the most virulent species: two isolates, Nemasys (LD₅₀=1.5–3.9) and Sus 94 (1.8–3.9) were superior to the rest.Steinernema kraussei (Steiner) was the least infectious nematode tested as its infection probability never exceeded 0.10. In addition, lethal dose₅₀ values for this species ranged from 9.0–62.6 and two isolates failed consistently to infect any hosts.     

Effect of nematode Panagrellus redivivus density on growth, survival, feed consumption and carcass composition of bighead carp Aristichthys nobilis (Richardson) larvae

The study aimed to determine the optimum density of free‐living nematodes in feeding bighead carp, Aristichthys nobilis, larvae. In the first experiment, carp stocked at 25 larvae L^?1 were fed varying levels of nematodes (50, 75, 100, 125 and 150 per ml) twice a day for 21 days from the start of exogenous feeding. Final body weight was significantly higher (P < 0.05) in larvae fed 125 and 150 nematodes per ml than in those fed 50 and 75 per ml, but survival was low (61.8 and 63.6%, respectively). Survival rate was highest in larvae fed 100 nematodes ml^?1 (81.3%). Carcass analysis showed that larvae fed 125 and 150 nematodes ml^?1 had significantly lower body protein and higher body lipid than those fed other nematode densities. Carcass ash was similar for larvae fed 50–100 nematodes ml^?1 but it decreased significantly at the higher nematode densities. Carp larvae in a subsequent experiment were given 50, 75 and 100 nematodes ml^?1 per feeding. Newly hatched Artemia was the control feed. Nematode consumption and growth of the larvae were determined. Larvae were sampled at intervals of 2–4 days and the nematodes in the gut were counted and measured. At each nematode density, the number of nematodes present in the gut of the larvae increased significantly with time. At each sampling day, the number of nematodes in the gut did not differ significantly among treatments (P > 0.05) although it tended to increase with nematode density at day 2 and day 4 but decrease at day 7 onward. The carp larvae consumed significantly shorter nematodes on day 2 and day 4 than on the succeeding sampling days regardless of nematode density. However, the length of nematodes in the gut of the larvae did not differ significantly among the nematode densities. The final body weight of larvae increased with increasing nematode density. The body weight of larvae fed 100 nematodes ml^?1 did not differ significantly from that of larvae given Artemia nauplii. Results show that bighead carp larvae should be fed 100 free‐living nematodes per ml at each feeding time.     

Substrate dependent larval development and emergence of the mushroom pests Lycoriella auripila and Megaselia halterata

Pasteurized, spawned, full-grown and immediately-cased full-grown compost were simultaneously exposed to natural populations of the mushroom pests Lycoriella auripila (Winnertz) (Diptera: Sciaridae) and Megaselia halterata (Wood) (Diptera: Phoridae). Different numbers of adults emerged from each of these composts. Highest numbers of L. auripila emerged from spawned and pasteurized compost whereas lowest numbers of L. auripila emerged from full-grown compost. the emergence from full-grown compost was delayed, which could be explained by the delayed development of the larvae in this type of compost. High numbers of M. halterata emerged from compost that was completely colonized by the mycelia of the edible white button mushroom Agaricus bisporus (Lange) Imbach. The immediate covering of the compost with a casing layer significantly lowered the numbers of emerging M. halterata flies. Compared with the emergence pattern from full-grown and immediately-cased full-grown compost, adult M. halterata showed a delayed pattern of emergence in spawned compost. Adult M. halterata did not emerge from pasteurized compost. The results of these experiments enabled us to improve the timing of the application of insect pathogenic nematodes in the control of the larvae of both insect pests.     

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.