For each concentration 10 adult individuals (5 males and 5 females) were individually inoculated by applying 1 μl of the suspension on the thorax with a pipette, resulting in expected exposure rates of 102, 103, 104, 105 and 106 conidia per individual. After inoculation each individual was put singly in a sealed medicine cup under moist conditions. After 24 h incubation

in the moist chambers, the wasps EPZ015666 in vitro were provided with a cotton wick soaked in 0.3 M sucrose as food, in a new medicine cup. The food was renewed weekly. The wasps were incubated in L:D 16:8 h and monitored daily for 14 days. Dead wasps were surface sterilized as described in Section 2.3 and transferred to moist chambers. A wasp was considered to be mycosed if mycelia protruded through the cuticle after death and subsequently formed distinctive conidiation. The experiment was repeated on four different occasions, each time with 10 individuals for each concentration and fungal isolate. The treatments were arranged in a completely randomized design in polystyrene boxes as for D. radicum. The experimental arena (‘patch’) consisted of a 55 mm petri dish (VWR, Sweden), containing a 50 mm filter paper (quality 1701, Munktell Filter AB, Sweden) and a 35 × 35 × 6 mm piece of turnip. Larvae of D.radicum, treated as described Selleckchem Ruxolitinib for the respective choice bioassays in Sections 2.5.1 and 2.5.2 below, were distributed on the turnip. The thickness of the turnip allowed

free probing access for the parasitoid, since the ovipositor length is 2.9 mm ( Brown and Anderson, 1998). Around the turnip 2 ml of dry vermiculite (2–5 mm) was evenly distributed. The petri dishes were sealed with parafilm and incubated for 18 h in darkness at 20 ± 1 °C for D. radicum larvae to establish in the turnip. The following day, the parafilm and lids were removed. For the

respective choice bioassay, the vermiculite was then inoculated as described below in Sections 2.5.1 and 2.5.2. Just before the onset of the choice bioassays a 20 mm high cylindrical aminophylline metal barrier (mesh width 0.8 mm, Sintab, Sweden) with 5 mm inward overhang was placed around the outside of each petri dish to prevent larvae from leaving the arena. Two arenas were placed inside a transparent plastic box (185 × 185 × 115 mm). At the onset of the choice bioassays, a 2–4 day old mated and sugar-fed female T.rapae was introduced into each box. The parasitoid had access to water and 0.3 M sucrose in 30 ml cups through a 4 cm piece of dental cotton roll throughout the experiment. The bioassays were performed in a climate cabinet for 24 h at 20 ± 1 °C and L:D 16:8 h with illumination provided by white fluorescent lamps (Long Life T8 Ultimate 36 W/830 3000 K, Aura Light, Sweden) reaching ca 4200 lux inside the boxes. After termination of the experiments described in Sections 2.5.1 and 2.5.2, the females of T. rapae were incubated individually in medicine cups at 20 ± 1 °C in L:D 16:8 h, provided with a cotton wick soaked in 0.