The neem leaf extract was prepared by crushing 100 g of neem leaves in water and soaking in water overnight; the neem seed kernel – V. negundo leaf extract was prepared by taking 100 g each neem seed kernel powder and V. negundo Selleck Osimertinib leaves. They are then crushed and soaked in water overnight and filtered before use for field trials. The 2nd, 3rd, 4th and 5th instar larvae
were grown in plastic containers covered by a muslin cloth for aeration. Each container consists of 10 larvae and three replicates were maintained. Ten milliliters of spore suspension of the fungi were taken in which each larva was dipped thoroughly for 10 s. The control larvae were dipped in 0.02% Tween 80 alone. The containers with larvae were maintained at 26 ± 1 °C temperature; relative humidity 70 ± 10% and photoperiod of 16:8 L:D. Larval mortality was recorded at every 24 h interval for seven days after treatment and the data was analyzed statistically. The cadavers were used for re-isolating the pathogen in pure culture for confirming the pathogenicity of fungi. The larvae were fed twice a day with a specially formulated diet (slightly modified diet of6) which see more consists of caesin-10 g, sucrose-20 g,
ascorbic acid-2 g, Brewer’s yeast-2 g, sorbic acid-0.65 g, formaldehyde-1 ml, agar-6 g, turmeric leaves-50 g and water-275 ml. The unfed feed and leaves were removed periodically. Field trials were conducted for two years at one of the turmeric farms in Karungalpalayam, Erode, Tamil Nadu, India during 2010–2011 in randomized complete block design having 11 treatments which includes an untreated control plot with three replicates for each treatment. Each treatment plot size was 10 m2 with 50 plants in each plot. Treatments were applied as foliar sprays and comprised as follows: T1 – M. anisopliae; T2 – B. bassiana; T3 – Standard N. rileyi (MTCC 4175); T4 – Standard H. citriformis (MTCC 6800); T5 – H. citriformis
HC28; T6 – N. rileyi NR07; T7 – Neem leaf extract; why T8 – Neem seed kernel + V. negundo leaf extract; T9 – Commercial Biopesticide (Biopower®); T10 – Acephate; T11 – Untreated control. The spraying of bioformulations was done using a Knapsack sprayer with a spray volume of 300 L ha−1. The treatment sprays were applied twice at two days interval. Soap powder (2 g/L) and/or starch powder was added to enhance the adhesiveness of the sprays as the whole experiments were conducted during rainy season.10 The observations were recorded on ten randomly selected plants in each plot. Data on the death of larval population after 3, 5 and 7 days after spraying were calculated.