formicarius. Pheromone lures consisting of rubber septa loaded with Z3-dodecenyl-E2-butenoate, sealed in an impermeable bag for shipping and storage, were obtained from Chem Tica Internacional S.A. (San José, Costa Rica). Pherocon unitraps (Trécé Incorporated, Adair, Oklahoma, USA) baited with these lures were used to trap adult C. formicarius in sweet potato fields in Latte Heights (Guam, USA) during 2010. The trapped adults were taken to the laboratory, placed in batches in collapsible cages (12 × 10 × 10 cm), fed leaves and pieces of the sweet potato,
and maintained at 22 ± 2 °C, 70–80% relative humidity and a 16:8 h L:D photoperiod. Approximately 5–6 generations were completed before using the offspring for experiments. For all experiments,
3–4 week old adults were obtained from these laboratory colonies ( Gadi and Reddy, 2014). Conidia of B. bassiana strain 17-AAG cost GHA were supplied as an unformulated technical grade powder by Laverlam International (Butte, Montana, USA). The conidial titer was 1.6 × 1011 conidia/g and viability was 98%, based on conidial germination in the laboratory on potato dextrose yeast extract agar after incubation for 18 h at 27 °C. Cultures of M. brunneum F52 (a commercialized isolate previously identified as M. anisopliae) were obtained from Novozymes Biologicals Inc. (Salem, Virginia, USA). Conidial powders were stored dry check details at 4–5 °C until formulation and use. The chemicals used in the present study – azadirachtin (Aza-Direct) and spinosad – were obtained as shown in Table 1. Laboratory tests were carried out from 12 September to 15 October 2013 with the hypothesis that the chemicals we tested, when topically applied, would exhibit contact toxicity to C. formicarius adults ( Table 1). For each replicate, 10 adults were transferred to a disk of Whatman No. 1 filter paper (9 cm diam, Whatman® quantitative
filter paper, ashless, Sigma–Aldrich, St. Louis, Missouri, USA) in a 9 cm disposable Petri dish. Each dish received a 10-g piece of sweet potato and two 7 cm sweet potato branches with leaves (4–8) as food for the insects. Five replicate (prepared at PDK4 separate times using different cultures and batches of insects) Petri dishes of 10 adults were sprayed (Household Sprayer, Do It Best Corp., Ft. Wayne, Indiana, USA) with 0.5 mL of its assigned treatment (Leng and Reddy, 2012). Two control treatments were maintained; in one, the dishes were sprayed with 0.5 mL of tap water, and in the other, no treatment was applied. Following applications, dishes were maintained under laboratory conditions (previously described), and adult mortality was assessed at 24, 48, 72–96, 120–144, and 168–192 h after treatment.