Virulence of Metarhizium brunneum (Ascomycota: Hypocreales) Strains Against Stinkbugs Euschistus heros and Dichelops furcatus (Hemiptera: Pentatomidae).

Three strains of fungi belonging to the genus Metarhizium Sorokin (ARSEF 4556, ARSEF 3297, native strain) were assayed against adults and nymphs of the Neotropical brown stinkbug Euschistus heros (F.) and the green-belly stinkbug Dichelops furcatus (F.). The most virulent strain, ARSEF 4556, caused over 90% mortality. The average survival time of the second and fifth instar nymphs and adults following immersion in 1 × 108 conidia ml-1 was 4.8, 5.7, and 5.2 d, respectively. The second instar nymphs were more susceptible than the adults. The LC50 values and median survival times for second instar and adult E. heros were 1.6 × 107 and 3.1 × 107 conidia ml-1 and 6 and 8 d, respectively. Eggs of E. heros and the closely related stinkbug, D. furcatus, were highly susceptible to ARSEF 4556 with the mean mortality of eggs immersed in 1 × 108 conidia ml-1 being 77.4% and 89.7%, respectively. The strain 3297 showed also good aptitudes for stinkbugs control with mortalities higher than 80% against nymphs and adults and eggs mortalities of 75.5% for E. heros and 79.6% for D. furcatus. This study has shown that it is possible to have a two-pronged control strategy, targeting adults and to reduce oviposition and targeting egg clusters to prevent emergence and dispersal of nymphs. Besides early instars of nymphs have been shown to be more susceptible to the fungal strains than late instars and adults.

Soil drenching with entomopathogenic fungi for control of the soil-dwelling life stages and adults of the same generation of Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae).

Four Beauveria bassiana and three Metarhizium brunneum isolates were evaluated, as soil drenches, against Spodoptera littoralis prepupae. Treatment efficacy was determined by assessing total mortality during development from prepupae through to pupae and adults; mortality and sub-lethal effects on reproduction were also quantified for adults emerging from surviving prepupae/pupae. All isolates were pathogenic but overall mortality varied between 31.7 and 83.3% (0% for control); average survival time was 7.5-10.5 days (14.0 days for control). From 1.7-15.0% of adults emerging from surviving prepupae/pupae were deformed (0% in control). Contact with fungal suspensions as prepupae/pupae caused a significant reduction in fecundity of emerging adult females (15-58.9%), and a significant reduction in egg viability (6.8-28.4%) compared with controls. Two isolates were selected for virulence evaluation against S. littoralis prepupae. The LC50s were 1.7 × 107 and 1.8 × 107 conidia ml-1 and the median survival times were 7 and 6 days for isolates EAMa 01/58-Su and EAMb 09/01-Su, respectively. Destruxin A was present in pupae developing from prepupae treated with isolates EAMa 01/58-Su (0.010 ± 0.002 µg pupae-1) and EAMb 09/01-Su (0.015 ± 0.003 µg pupae-1). The use of entomopathogenic fungi as soil drenches could be a key component of S. littoralis IPM strategies due to direct reductions in the number of soil-dwelling life stages and, also, the significant reduction in reproductive potential of surviving adults.

Transient endophytic colonizations of plants improve the outcome of foliar applications of mycoinsecticides against chewing insects.

The current work reports how spray application of entomopathogenic fungi on alfalfa, tomato and melon plants may cause an additional Spodoptera littoralis larvae mortality due to a temporal colonization of the leaves and subsequent ingestion of those leaves by the larvae. Most entomopathogenic fungi (EF) (Ascomycota: Hypocreales) endophytes seem to colonize their host plants in a non-systemic pattern, in which case at least a transient endophytic establishment of the fungus should be expected in treated areas after spray application. In this work, all strains were able to endophytically colonize roots, stems and leaves during the first 96h after inoculation. Whilst the treatment of S. littoralis larvae with a 10(8)ml(-1) conidial suspension resulted in moderate to high mortality rates for the Metarhizium brunneum EAMb 09/01-Su (41.7-50.0%) and Beauveria bassiana EABb 01/33-Su (66.7-76.6%) strains, respectively, an additive effect was detected when these larvae were also fed endophytically colonized alfalfa, tomato, and melon leaves, with mortality rates varying from 25.0% to 46.7% as a function of the host plant and total mortality rates in the combined treatment of 75-80% and 33-60% for B. bassiana and M. brunneum, respectively. Fungal outgrowth was not detected in any of the dead larvae feeding on colonized leaves, whereas traces of destruxin A were detected in 11% of the insects fed tomato discs endophytically colonized by M. brunneum. The combined effects of the fungal spray with the mortality caused by the feeding of insects on transient EF-colonized leaves have to be considered to estimate the real acute impact of field sprays with entomopathogenic fungi on chewing insects.