Field Infection of Zonocerus variegatus following application of an oil‐based formulation of Metarhizium flavoviride conidia

Conidia of Metarhizium flavoviride grown in diphasic culture with sporulation on rice, were extracted in kerosene and formulated with peanut oil. The formulation led to high mortality when applied to field populations of Zonocerus variegatus; between 70 and 95% of insects in samples collected from the field 0, 3 or 7 days after spraying died. Sporulation of M. flavoviride was observed in 70–80% of the dead‐treated insects, which compares favourably with results obtained in laboratory bioassays. Many dead Z. variegatus, some with internal sporulation, were found in shaded niches and oviposition sites in the treated plots. No sporulation was observed in samples collected prior to application, and both sporulation and mortality differed significantly between control and treated plots. Some infections were observed in insects collected 16 and 23 days after treatment; these late infections may be a consequence of contact with spores surviving in the environment.     

Effects of the Sunscreen Oxybenzone on Field Efficacy and Persistence of Metarhizium flavoviride Conidia Against Kraussella amabile (Orthoptera: Acrididae) in Mali,West Africa

A field trial was performed at Mourdiah, northwest Mali, to evaluate whether the performance of Metarhizium flavoviride conidia would improve if 2% oxybenzone, an oil-soluble ultraviolet light protectant, was used. Conidia were applied in an ultra-low volume formulation against nymphs of the grasshopper Kraussella amabile . Mortality was assessed by collecting grasshoppers directly from field plots (sweep net samples) or by exposing untreated nymphs to sprayed vegetation for 48 h (field cage samples). For sweep net samples, the mortalities in M. flavoviride treatments varied between 94.5 and 99.9% up to 7 days after treatment, while control mortalities were significantly lower (20.7-55.0%). There were no significant differences between M. flavoviride treatments with or without oxybenzone. The average survival times were found to be significantly lower for fungal treatments (4.9 days) compared with the control treatment (11.8 days). W ith field cage samples, the mortality in M. flavoviride treatments declined from > 95.0% for nymphs exposed just after application to < 30.0% for nymphs exposed to treated vegetation on the 10th day after application. The control mortalities varied between 5.1 and 37.2%, and were significantly lower than fungal treatments for samples placed just after application and 10 days after application. The overall average survival times were 10.9 and 10.7 days for M. flavoviride with and without oxybenzone respectively, and 13.9 days with the control treatment. An exponential regression was fitted using cage mortality data which indicated that the half-life for conidial infectivity was 4.3 days on treated vegetation.     

Peripheral-zone treatments for plum curculio management: validation in commercial apple orchards

A field trial was conducted using an oil formulation of the entomopathogenic fungus, Metarhizium flavoviride Gams & Rozyspal (Deuteromycotina: Hyphomycetes), to control the variegated grasshopper, Zonocerus variegatus L. (Orthoptera: Pyrgomorphidae). The influence of dispersal and secondary pick-up from the spray residue on mortality was assessed by sampling insects from the treated plots at intervals and monitoring disease levels during subsequent incubation in the laboratory. The cumulative mortality curves showed the pathogen to be highly effective, even in samples taken 8 days after spraying. A model was developed to link the influence of mortality, incubation, secondary pick-up of spores and insect dispersal, on the shape of these cumulative mortality curves. The model proved useful for data received from an experiment using small plot sizes, where insect dispersal is a big problem. The spray residue had an important influence on overall insect mortality. The model also indicated a difference in speed of kill between field and laboratory samples, suggesting an incubation delay in the field.     

Comparison of an Organophosphate Insecticide with a Mycoinsecticide for the Control of Oedaleus senegalensis (Orthoptera: Acrididae) and Other Sahelian Grasshoppers at an Operational Scale

Operational scale field trials were conducted in 1996 and 1997, in the east of the Niger Republic, on 50 and 800 hectare plots, to compare the efficacy of an oil based formulation of the entomopathogenic fungus, Metarhizium anisopliae (flavoviride) var. acridum (Deuteromycotina: Hyphomycetes) with fenitrothion for the control of Sahelian grasshoppers. The Senegalese Grasshopper Oedaleus senegalensis Krauss was the most abundant species in the trials. M. anisopliae was applied at 5 x 1012 spores ha-1 at volume application rates of 2 and 0.5 l ha-1 in successive years. Fenitrothion was applied at 220 g/ha-1 at 1.25 and 0.22 l ha-1 volume application rates. Ultra low volume equipment mounted on a vehicle (1996) or a fixed wing aircraft (1997) was used for application. The M. anisopliae treatment reduced the grasshopper population significantly after 7 days and by 93% within 16 days. Fenitrothion caused a population reduction of more than 90% shortly after application, but due to immigration, the grasshopper population recovered to the initial level within 16 days. Grasshoppers treated with the fungus and given the opportunity to thermoregulate in the sun died more slowly than grasshoppers incubated in the shade. The survival of spores in the spray residue of the M. anisopliae plots assessed by exposing grasshoppers to the sprayed vegetation at intervals and monitoring disease levels during subsequent laboratory incubation, showed the spray residue to remain highly infective, for three weeks after spraying. At the end of the 1997 season, egg pod density and viability in the plot treated with the fungus was reduced compared with both untreated and the fenitrothion plots. Compared with the existing practice of large-scale treatment of grasshopper infestations with fenitrothion, use of M. anisopliae would not only be safer to mammals and less damaging to non-target organisms, but also be more effective in the long-term control of grasshoppers.     

Field Treatment of Desert Locust ( Schistocerca gregaria Forskal) Hoppers in Mauritania Using an Oil Formulation of the Entomopathogenic Fungus Metarhizium flavoviride

A field trial was conducted to test the efficacy of an oil formulation of the entomopathogenic fungus , Metarhizium flavoviride Gams & Rozsypal (Deuteromycotina: Hyphomycetes) , as a control agent for the desert locust , Schistocerca gregaria Forskal (Orthoptera: Acrididae) , in Mauritania . The treatment was applied to hopper bands in the field . In caged samples of the treated insects , 99% mortality was observed in 15 days . On average during application only 51% of the hoppers received spray droplets directly , the additional mortality being due to pick - up of spores from the spray residue . This mortality was assessed by exposing insects to a treated plot at intervals , and monitoring disease levels during subsequent incubation in cages . The spray residue remained highly infective for a period of 6 days after application . The number of nymphs in the hopper bands was estimated using a computer - based photographic image - processing technique . Unfortunately , most of the hopper bands dispersed into small groups , but a population reduction could be demonstrated for one hopper band . Maximal daily mortality in the field was observed 10 - 11 days after application , compared with 6 - 10 days in cages .     

Reduction of Feeding by the Variegated Grasshopper, Zonocerus variegatus , Following Infection by the Fungal Pathogen, Metarhizium flavoviride

The effect of infection by the fungal entomopathogen , Metarhizium flavoviride, on feeding by the tropical grasshopper pest , Zonocerus variegatus, was investigated in field - cage studies . A significant reduction in feeding , as indicated by faecal production , was recorded 2 - 3 days after inoculation for a range of spore doses (104 , 105 and 5 105 spores per insect) . This was before any mortality was recorded due to infection . All infected individuals died by day 7 . At this time , faecal production of the treated grasshoppers was equivalent to less than 2 days faecal production by grasshoppers untreated with spores . This reduction in feeding (69 , 71 and 74% total reduction by day 7 in the 104 , 105 and 5 105 doses respectively in comparison to controls) is a substantial contribution to the overall effects of the slow acting pathogen . Furthermore , the rapid reduction in feeding indicated that this effect was not simply due to invasion of the host tissues by the pathogen or production of secondary metabolites . The possibility that reduction in feeding is associated with a behavioural response in which there is a trade - off between host defence and feeding during early stages of infection is discussed     

Effect of organic and mineral fertilizers on N-use by wheat under different irrigation frequencies

A field trial was established in Errachidia, southern Morocco, to investigate the interaction between wheat residue management and mineral 15N-labelled ammonium sulphate, under different irrigation treatments, applied to wheat (Triticum durum var. Karim). In treatments I1, I2, I3 and I4, plots were irrigated every 10, 15, 21 and 30 days. Each plot contained three sub-plots that received three fertilization treatments: T1 received 42 kg N ha-1 of ammonium sulphate before seedling, 42 kg N ha-1 of ammonium sulphate labelled with 9.764 at % 15N excess at tillering and 84 N kg ha-1 of ammonium sulphate at flowering; T2 received 42 kg N ha-1 of ammonium sulphate labelled with 9.764 at % 15N excess at seedling, 42 kg N ha-1 at tillering and 42 kg N ha-1 at flowering; T3 received 4800 kg ha-1 of wheat residue labelled with 1.504 at % 15N excess and 42 kg N ha-1 of ammonium sulphate before seedling and 42 kg N ha-1 of ammonium sulphate at flowering. Nitrogen fertilization with 168 kg N ha-1 did no significantly increase grain and straw yields in comparison to the 126 kg N ha-1 application. The combination of the organic input and supplementary application of mineral fertilizer N has been found as a more attractive management option. For all irrigation treatments, the % recovery of N in the whole plant was higher in plants that received 15N at tillering (63%, 49% respectively for irrigation intervals between 10 and 30 d) than in plants that received 15N just after seeding (28% for irrigation each 10- and 30-d intervals). For the irrigation treatment each 10 and 15 days, the 15N was mainly recovered by the grain for all fertilization treatments, whereas for irrigation treatment each 30 days, the grain and straw recovered nearly equal amounts of fertilizer. For grain and straw of wheat, nitrogen in the plant derived from the fertilizer was low, while most of the N was derived from the soil for all irrigation and fertilization treatments. The % nitrogen in the plant derived from the fertilizer values showed no significant difference between the different plant parts. The results suggested a dominant influence of moisture availability on the fertilizer N uptake by wheat. Under dry conditions the losses of N can be allotted to denitrification and volatilisation.     

Application of Metarhizium flavoviride (Deuteromycotina: Hyphomycetes) Spores Against the Tree Locust, Anacridium melanorhodon (Orthoptera: Acrididae), in Sudan

Spores of the entomopathogenic fungus Metarhizium flavoviride were applied against the tree locust Anacridium melanorhodon melanorhodon in the field for the first time. The treated areas covered a total of 55 ha in the vicinity of Tendelti in the White Nile State of Sudan. At a dose of 100 g of spores/ha applied in 5 l of oil formulation, the application led to a reduction in population densities of 66-76% after 18 days. Correction of the data with the Henderson/ Tilton formula gave an overall control efficiency of 68%.     

Effet de l'application d'un mélange lambda-cyhalothrin (pesticide chimique) et de spores de Metarhizium anisopliae (flavoviride) var. acridum Driver and Milner (biopesticide) appliqué sur les larves de sauteriaux au Mali

A mixture of lambda-cyhalothrin (lambda-cyhalothrin: chemical insecticide) and Metarhizium anisopliae ( flavoviride ) var. acridum Driver and Milner, an entomopathogenic fungus (bioinsecticide) was used for grasshopper control in Mali. An oil-based formulation of Metarhizium anisopliae ( flavoviride ) var. acridum Driver and Milner has been developed by LUBILOSA a collaborative project for locust and grasshoppers control. It takes 6 to 10 days for the biopesticide to kill the hosts, which is not a problem for larvae in fallows because they will die before reaching the farmers' fields. However, if crops are infested by adults, the farmers can not wait for 6 to 10 days. An experiment was conducted in Mali using a mixture of a biopesticide and chemical pesticide. The mixture of lambda-cyhalothrin (chemical insecticide) and Metarhizium anisopliae ( flavoviride ) var. acridum (biopesticide: oil-based entomopathogenic fungus spore suspension) was applied to nymphs of Sahelian grasshoppers, using ultra low volume (ULV) sprayers. Both the mixture and lambda-cyhalothrin alone gave quick mortality, with slightly higher mortality for the mixture. Mortality due to the Metarhizium treatments began 2 days after application and subsequently reached similar levels of mortality to the lambda-cyhalothrin mixture treatments. The efficacy of the mixture was greater than Metarhizium alone. The efficacy of lambda-cyhalothrin reached 80% on the day following application, but declined after 10 days, due probably to immigration of untreated grasshoppers.     

Efficacy of Beauveria bassiana (Bals.) Vuill. against the tarnished plant bug, Lygus lineolaris L., in strawberries

Beauveria bassiana has a high insecticidal potential to control the tarnished plant bug, Lygus lineolaris, a significant pest of strawberries. Screening experiments showed that L. lineolaris adults were susceptible to several B. bassiana isolates. Another screening test with Coleomegilla maculata, a natural enemy found in strawberries, was also performed in order to select the isolate having lower entomopathogenic impact on this insect. Based on data obtained from both insect species and on the ecozone origin of the B. bassiana isolates, INRS‐IP and INRS‐CFL isolates were selected for further experiments. The LC₅₀ values of these two isolates against L. lineolaris adults were 7.8 × 10⁵ and 5.3 × 10⁵ conidia/ml, and average survival time (AST) values were 4.46 and 4.37 days at a concentration of 1 × 10⁸ conidia/ml respectively. Results also indicated that L. lineolaris nymphs are susceptible to the selected isolates. During field experiments, using a randomized block design with four replicates, INRS‐IP and INRS‐CFL isolates were applied at two rates (1 × 10¹¹ and 1 × 10¹³ conidia/ha) weekly during a period of 4 weeks. These multiple applications triggered a significant reduction of L. lineolaris nymphal populations in strawberries. Twenty‐four days after the first application, a significant difference was observed between the mean population densities of surviving nymphs in all B. bassiana‐treated plots (less than one insect per five plants) compared with those in control plots (four insects per five plants). During the field experiment, persistence of insecticidal activity and viability of B. bassiana conidia were also monitored. The results showed the presence of viable and infective conidia up to 6 days after each application on strawberry foliage. Moreover, the multiple applications of B. bassiana at the rate of 1 × 10¹³ conidia/ha triggered a significant reduction in strawberry fruit injuries induced by L. lineolaris feeding behaviour compared with the control plots.     

Detection of molecular variation in the insect pathogenic fungus Metarhizium using RAPD-PCR

Abstract DNA polymorphism among isolates of the insect pathogenic fungus Metarhizium anisopliae and M. flavoviride was investigated by RAPD-PCR. DNA fragments of between 0.3 and 2.7 kb were obtained using eight 10-mer PCR primers of arbitrary nucleotide sequence, and each isolate differed in the size and number of RAPD products, indicating considerable polymorphism. Isolate-specific RAPD fingerprints were used to calculate relative genetic similarity; this differentiated isolates into two major groups, separating nine of the ten isolates of M. anisopliae from the two of M. flavoviride . However, an Australian M. anisopliae isolated from an Orthopteran host exhibited a higher degree of genetic similarity to the M. flavoviride group. M. anisopliae isolates were further segregated into three subgroups which were loosely related to their geographical origins. although considerable polymorphism was observed within these groups. There was no apparent association between genotype and original insect host.     

Reduction of grasshopper populations following field application of the fungus Beauveria bassiana

Conidia of the Hyphomycete fungus Beauveria bassiana (Bals.) were applied in an attempt to reduce field populations of grasshoppers, primarily the migratory grasshopper Melanoplus sanguinipes (Fabricius). Dry spores were applied with wheat bran carrier to three fallow fields at a rate of 2.0 × 10¹³ spores ha^?1 in 10 kg bait ha^?1. Examination of culture plates that had been placed in the field to capture spores and of bran carrier with scanning electron microscopy indicated that a substantial portion of the B. bassiana colony—forming units (spores and clumps of spores) did not adhere to the bran and were applied in the field as free particles. Grasshoppers collected from the treated plots at intervals after application were assayed for infection by B. bassiana. The observed rate of mycosis in the treated populations was 70% of those collected after 2 days, declining to 41% by 13 days and 5% by 19 days after application. Analysis of reductions in population density gave results in agreement with the infection data. Treated populations declined 60% and 33% by 9 and 15 days after application respectively. The reductions were significant on both post—treatment sampling dates (p < 0.05) and the three replicated fields gave comparable results. This is the first field demonstration of effectiveness of this fungus as a microbial control agent of grasshoppers.     

Laboratory and Field Trials of Bait Formulations of the Fungal Pathogen, Metarhizium flavoviride, Against a Tropical Grasshopper and Locust

Extrusion technology was used to produce maize-starch 'contact bait' carriers, and vegetable-oil suspensions of Metarhizium flavoviride were incorporated into the baits after extrusion. Laboratory bioassays using the locust, Schistocerca gregaria, demonstrated mortality and reduction of 8 1 feeding after exposure to the baits, and gave an LC 50 of 1.5 10 spores l slope 0.63 for fifth instar nymphs. In laboratory trials, maize-starch contact baits showed improved efficacy relative to wheat-bran edible bait formulations of M. flavoviride. Field cage experiments, undertaken in Mali, using fourth and fifth instar nymphs of the grasshopper, Hieroglyphus dagenensis, demonstrated significant mortality with both fresh and weathered maize-starch contact bait formulations of M. flavoviride, relative to untreated controls.     

Studies on insecticidal potential of extracts of Gmelina arborea L. products for insect pests control on cowpea. 1. The legume flower bud thrips,Megalurothrips sjostedti Trybom

Abstract

The effects of weekly foliar application of extracts of Gmelina arborea (L.) fruits, bark and leaves on flower bud thrips, Megalurothrips sjostedti Trybom and the pod carrying capacity of cowpea variety SAMPEA 7, were investigated in field studies in the 1998 and 1999 cropping seasons. All the Gmelina extracts and the synthetic insecticide (Uppercott) caused significant a reduction of thrips on treated plots compared to the untreated check. However, no significant differences were observed on the number of thrips after the first two weeks of spraying and insect sampling between the fruits, bark extracts on one hand and Uppercott on the other in both seasons. This period corresponds to the period of high thrips number on this variety of cowpea planted in the northern Guinea savanna ecological zone of Nigeria. Pod density (a measure of the ability of insecticides to control the population of thrips on field cowpea) observed in untreated plots was inferior to those in treated plots. Uppercott treated plots had superior pod density compared to plots sprayed with Gmelina extracts while the fruit extract produced superior number of pods among the other extracts treated plots.     

Implications of pre- and post-fertilizing changes in growth and nitrogen pools following multiple applications of nitrogen fertilizer to a Pinus radiata stand over 12 years

Around the world large tracts of forest, previously available for production, have been reserved for nature conservation. This means that wood supply must be met from a reduced land base, including land of low productivity. In addition there are likely to be increasing demands on the use of managed forests for sequestering C as one means of reducing the build up of atmospheric CO2. One way for the forest industry to meet the demands of increased production would be through the use of fertilizers. Substantially increased growth from fertilizer N application has been measured in many cases while, in the northern hemisphere, atmospheric N deposition has been associated with increased growth in some forests.The possibility of using fertilizer N to increase growth, and the effect on the forest and soil, was studied in a research trial area in north-east Tasmania, Australia. Nitrogen was applied for 12 years to a 16 year old P. radiata stand in a low rainfall zone. Growth and foliar nutrient concentrations were measured to age 34 years, to determine change after attainment of steady state growth at age 25 years, and following cessation of fertilizing at age 29 years. Biomass sampling was carried out at ages 25 years, 29 years, and 34 years.Growth at the steady peak rate achieved in the fertilizer plots, of 31 m3 ha-1 periodic annual increment, was accompanied by changes in the N nutrient pools. Surface applied N built up in the surface litter layer while this layer was increasing, from 15 t ha-1 to nearly 50 t ha-1, and fertilizer was being applied between ages 25 and 29 years. Decline of total N in the soil, between ages 25 and 29 years, indicated continued uptake from that source even though fertilizer was being applied. Subsequent increases in soil N, through leaching from the litter layer, followed cessation of fertilizing.Following cessation of annual applications of N fertilizer, growth rates declined by less than 15% and remained substantially (>150%) above the control growth rates for 4 years. Over a 3 year period foliar N concentrations declined steadily, to the same as control plots, although mass of needles was not reduced 5 years after cessation of fertilizer applications and remained significantly above that of the control plots.Without fertilizers, N for new foliage must be supplied from internal recycling and from mineralization of litter and soil organic matter. There were net losses of N from the foliage and the wood, estimated at around 12 kg N ha-1 yr-1, for the fertilized trees for the 5 years following cessation of fertilizing. This indicated retention and recycling within the tree of a considerable proportion of the 104 kg N ha-1 in the foliage, at age 29 years. As rates of mineralization of soil N were likely to be low, this retention of N within the tree was important in maintaining growth rates.In the forest ecosystems investigated, with low initial content of soil organic matter and N, fertilizer N produced improved tree health and substantially increased growth, thus providing the opportunity to manage this forest for increased wood production or C sequestration. Although there was a substantial build up of N and C in the litter on the fertilized treatment, incorporation of N and C into the mineral soil was slow. Over 12 years 1.34 t N ha-1 had been applied, comparable to 40 years of atmospheric deposition at 30 kg N ha-1. There were no signs of detrimental effects from this application.     

The Susceptibility of Boophilus annulatus (Ixodidae) Ticks to Entomopathogenic Fungi

The pathogenicity of five species of entomopathogenic fungi (Deuteromycetes, species: Beauveria bassiana, Metarhizium anisopliae, Metarhizium flavoviride, Paecilomyces fumosoroseus and Verticillium lecanii ) to the various developmental stages of Boophilus annulatus ticks was compared under laboratory conditions. M. anisopliae and B. bassiana strains were most virulent to engorged females and caused 85-100% mortality within 7-10 days post-inoculation (PI). The highest mortality of engorged females caused by other fungi reached only 25-60%. All tested fungi prevented or reduced the egg laying capability of the ticks several days before their death. Females surviving after treatment with the most virulent M. anisopliae strain (Ma-7) reached only 7-8% of their egg laying capacity as compared with the control. Other fungi caused a reduction of the weight of laid eggs by 35.4-80.8% as compared with untreated females. Only M. anisopliae and B. bassiana strains caused 70-98% mortality of the treated eggs. Unfed larvae of Boophilus annulatus were sensitive to M. anisopliae and M. flavoviride strains. The Ma-7 strain was most virulent to unfed larvae, with a mortality rate of 80.4% at a concentration of 1 ×107 spores ml -1 and 100% mortality at a concentration of 1 ×108 spores ml -1 .     

Molecular characterization of isolates of Metarhizium from locusts and grasshoppers

The relationships between 30 strains of Metarhizium were investigated by isoenzyme analysis, PCR-RAPDs, and protease production. The strains included representatives of M. anisopliae, M. flavoviride var. flavoviride and M. flavoviride var. minus. Thirteen isolates conforming to M. flavoviride from acridoid hosts and pathogenic to locusts and grasshoppers were shown to be a single, distinctive genotype with a pan-tropical distribution. In addition, the separation of M. flavoviride var. minus as a host specific taxon distinct from European isolates of M. flavoviride was also supported. The possibility of coevolution within some insect pathogenic populations is discussed.     

Field Trial with the Entomopathogenic Fungus Metarhizium anisopliae var. acridum against Bands of the Grasshopper Rhammatocerus schistocercoides in Brazil

The efficacy of a mycoinsecticide formulated in vegetable oil was tested in Brazil against the grasshopper Rhammatocerus schistocercoides . A set of experiments was conducted in the Chapada dos Parecis region (Mato Grosso state), a permanent zone of outbreaks for this pest. Experiments were performed in zones of natural vegetation, against grasshopper bands in the third nymphal instar. Three nymphal bands were treated with a mycoinsecticide formulation based on conidia of the entomopathogenic fungus Metarhizium anisopliae var. acridum ( =M. flavoviride ), strain CG 423. Three non-treated bands were used as control. The application was made with the aid of a hand-held ULV sprayer adjusted to deliver 2 l of the formulation ha -1 , each containing 1 ×10 13 conidia. Treatments were limited to the surface of the grasshopper bands and their immediate borders (5-10 m). The efficacy of the mycoinsecticide was evaluated through band survival after treatment (grasshopper numbers, surface, density, behaviour and daily movement of the band), allowing the insects to move freely in their natural environment. Insects were regularly surveyed and maintained in the laboratory, allowing estimates of the infection rate. Field and laboratory studies showed a clear effect of the product 10 days after treatment. At 14 days post-spraying, mortality caused by the mycoinsecticide in the field was approximately 88%.     

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

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

Effect of different isolates of Beauveria bassiana on field populations of Lygus hesperus

Lygus hesperus (Knight) (Hemiptera: Miridae) is a particularly damaging pest of many crops in the Western United States. Current control tactics are chemically based and there is some concern over resistance building up in populations. Based on previous laboratory studies conducted in California and Mississippi, USA, two new isolates of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Deuteromycotina: Hyphomycetes) were selected for field-testing against L. hesperus in California. Alfalfa plots were treated with one of three isolates of B. bassiana (a commercial isolate, an isolate from CA (WTPB2) or an isolate from MS (TPB3)) or the chemical pesticide Warrior T. More than 75% of the adults collected from plots 3 days after application with B. bassiana were infected but no differences in percentage infection occurred among fungal treatments. In addition, approximately 30% of the insects collected from control plots or plots treated with Warrior T were also infected. PCR analysis using SSR markers revealed that the isolate causing most of the infections in fungus treated plots was the isolate applied. A mix of infections was found in control plots and plots treated with Warrior T. Despite high levels of infection, no significant reductions of adult populations occurred until 10–14 days after application when plots treated with Warrior T or B. bassiana had about half the numbers of adult L. hesperus as the control plots.