IOBC/wprs Bulletin Vol. 28(2) 2005

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grown in the dark for 3 weeks at 22˚C. Conidia were harvested by washing the plates with 0.1% Tween 80 and resulting suspensions were diluted to a concentration of 10 7 conidia per ml. Forty ml of each isolate suspension were combined and mixed with 4kg of Beauveria free soil by repeated cycles of spraying of the suspension on the soil (hand spray applicator) and subsequent mixing. This resulted in a soil sample containing a conidia concentration of 10 5 /g soil for each isolate. The concentration in the soil was verified by applying the SM method and counting colony forming units. Re-isolation of B. brongniartii from the soil was performed by applying the three isolation techniques each in 40 repetitions. For each isolation 30g soil were used. Baiting with M. melolontha (MB) was performed according to the GB method. Eight days post inoculation Melolontha and Galleria larvae were moved to new plastic cups (4.5 cm ∅, 6 cm high) filled with damped peat and further incubated in constant darkness at 22˚C. B. brongniartii strains were re-isolated from diseased larvae up to 70d postinoculation for MB and 30d for GB. Per diseased larvae one isolate was obtained and subjected to single colony isolation on CM. For re-isolations with the SM method one single colony per SM plate was isolated. Numbers of obtained isolates per isolation technique are listed in Table 2. The genotype of each isolate was determined by applying microsatellite analysis as above. Table 1. Genotypes of selected B. brongniartii isolates Isolate No. Original isolation technique Microsatellite loci and allele sizes [bp] Bb1F4 Bb2A3 Bb2F8 Bb4H9 Bb5F4 Bb8D6 Genotype 1 MB 214 106 196 171 154 172 A 4 MB 214 106 208 165 154 172 B 12 GB 196 103 181 177 157 172 C 19 GB 238 124 217 180 199 172 D 22 SM 238 127 211 177 208 172 E 29 SM 214 106 193 165 151 172 F Results and discussion For each isolation technique 10 genetically unique isolates, which were selected with one particular technique only, were tested for their virulence against M. melolontha larvae. Results of the bioassays are shown in Figure 1. The average survival time varied across all isolates from 24d to 62d. The average of the average survival times was 39.6d ± 8.6d for isolates obtained with MB method, 42.9d ± 9.2d for isolates obtained with GB method and 36.2d ± 5.0d for isolates obtained with SM method. Non of the averages differed significantly among the isolation techniques. These results suggest that the three techniques do not select for isolates with different virulence against M. melolontha. Investigations of various growth characteristics such as in vitro biomass, blastospore or oosporein production supported these results (data not shown). No significant differences among averages of the according parameters of the three isolate groups were found (data not shown). 27
<strong>28</strong> Average survival time [d] 70 60 50 40 30 20 10 0 Figure 1. Virulence against M. melolontha of 30 B. brongniartii isolates obtained either by the MB ( ), GB ( ), or SM ( ) method Table 2. Abundance of re-isolated genotypes Re-isolation technique 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 <strong>28</strong> 29 30 Isolate no. No. of isolates Genotype Original isolation technique Abundance of genotype Observed Expected MB 18 A MB 3 (16.7) a 3.0 B MB 7 (38.9) 3.0 C GB 0 ( 0.0) 3.0 D GB 4 (22.2) 3.0 E SM 3 (16.7) 3.0 F SM 1 ( 5.6) 3.0 GB 25 A MB 3 (12.0) 4.2 B MB 3 (12.0) 4.2 C GB 4 (16.0) 4.2 D GB 11 (44.0) 4.2 E SM 1 ( 4.0) 4.2 F SM 3 (12.0) 4.2 SM 40 A MB 9 (22.5) 6.7 B MB 11 (27.5) 6.7 C GB 7 (17.5) 6.7 D GB 11 (27.5) 6.7 E SM 0 ( 0.0) 6.7 F SM 2 ( 5.0) 6.7 a Relative observed abundance of genotypes among isolates re-isolated with the same technique To test whether the different techniques select certain genotypes or whether they equally selected different genotypes, two isolates of each isolate group were mixed in equal amounts with a soil sample. Subsequently, isolates were re-isolated with the three isolation techniques and the re-isolation rate for each genotype was determined. Eighteen isolates were obtained with the MB method, 25 with the GB method and 40 with the SM method (Table 2.). Four
Page 1 and 2: IOBC / WPRS Working group „Insect
Page 3: Preface The Working Group “Integr
Page 6 and 7: iv ERICSSON Jerry University of Bri
Page 8 and 9: vi PÁZMÁNDI Christian Centre for
Page 10 and 11: viii
Page 12 and 13: x Metarhizium anisopliae for white
Page 14 and 15: xii
Page 16 and 17: Insect Pathogens and Insect Parasit
Page 18 and 19: stands (Levine et al. 2002) to feed
Page 20 and 21: In conclusion, the chemical control
Page 22: Kuhlmann, U. & W.A.C.M. Burgt 1998:
Page 25 and 26: 10 Austria, Belgium, Czech Republic
Page 27 and 28: 12 The control strategies used agai
Page 29 and 30: 14 1) Case study: Soil application
Page 31 and 32: 16 2) Case study: Genetic diversity
Page 33 and 34: 18 Bidochka, M.J., Menzies, F.V. &
Page 38 and 39: Results and discussion The first co
Page 40 and 41: Isolation of Beauveria brongniartii
Page 44: genotypes (A, B, D, and F) were re-
Page 47 and 48: 32 Since 1974 the financial losses
Page 49 and 50: 34 there is no more a strict three
Page 54 and 55: Numbers of captured beetles were an
Page 56 and 57: Mean captues +/- SE per trap & day
Page 58 and 59: dilution for plating (see above). F
Page 60: 45 “New” white grubs
Page 63 and 64: 48 The goal of this study was to de
Page 65 and 66: 50 In spring 2003 the density of M.
Page 67 and 68: 52 (Germany). The first treatment w
Page 69 and 70: 54 Application timing of H. bacteri
Page 74 and 75: Results and discussion Collection o
Page 76: Discussion During this study the en
Page 79 and 80: 64 Materials and methods Source of
Page 81 and 82: 66 100 80 60 40 20 0 100 80 60 40 2
Page 83 and 84: 68 During this study, selectivity a
Page 86: 71 Wireworms
Page 89 and 90: 74 Solanum tuberosum L. (Solanaceae
Page 91 and 92: 76 % CATEGORIZATION OF MORBIDITY FI
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78 of adults caught can reliably pr
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Insect Pathogens and Insect Parasit
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Results and discussion Before spray
Page 100:
References Parker, W.E. & Howard, J
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88 insecticide residues. Nonetheles
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90 References Cockbill, G.F., Hende
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92 Results and discussion a. Taxono
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94 published; some information has
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96 C2a: no sensitive crops in the s
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98 Furlan, L., Tóth, M., Ujvary, I
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100 Roberts, A.W.R. 1928: On the li
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102 Results and discussion Crop rot
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104 % damage by wireworms 100 80 60
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106 noticed that grass-clover in cr
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108 % damage by wireworms 60 50 40
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110 Materials and methods Soil Soil
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112 Emigration - deterrence of wire
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114 Table 1. The effect of broadcas
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116
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118 presence and abundance of wirew
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120 Furthermore, A. obscurus larvae
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122 Horton, D.R. & Landolt, P.J. 20
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124 Bait trapping 48 bait traps wer
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126 Acknowledgements We thank Anton
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128 history can be obtained (Scheu
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130 lower left hand corner of fig.
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132 Eggers, T. & Jones, T.H. 2000:
Page 149 and 150:
134 pheromone combinations for each
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136 Fig. 2. Click beetle spp. captu
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138 Fig 4. Click beetle spp. captur
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140 Fig 6. Click beetle spp. captur
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142 Siirde, K., Lääts, K., Erm, A
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Page 162 and 163:
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Fig. 2. Comparison of several trap
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Fig. 5. Catches of WCR in VARs+ tra
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Table 1. Performance characteristic
Page 170:
Miscellaneous
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158 Material and methods Field tria
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160 Figure 2: Abundances of Collemb
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162
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164 but also when used against the
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166 based EPN product in this exper
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168
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170 branches. Beetles can walk up t
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172 1999/ 2000, I 1999/ 2000, C 200
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174
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176 Experimental design During the
Page 193 and 194:
178 Acknowledgements We thank Stefa
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180 Spore suspension Conidia of M.
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182 Data about the stability of spo
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184 dence of host presence and appl
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186 Material and methods All habita
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188 Hajek, A.E. & Leger, R. J. St.
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190 Risk Assessment of Fungal Biolo
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192 It is scheduled that our future
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IOBC/wprs Bulletin Vol. 28(2) 2005

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