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There is no sampling methodology defined for the whitefly Bemisia tabaci MEAM1 (Hemiptera: Aleyrodidae) in soybean crops. We optimized a plan, through the minimization of total sampling variance, to evaluate population density of various whitefly stages, based on a feasible sample size. The sampling plan proposed for N1–N4 immature stages consists of four random sampling points in a field. Per point, four leaflets are collected per third of a plant's height (one leaflet per plant), evaluating 1 cm2 per leaflet.

The use of Metarhizium against sugarcane spittlebugs in Brazil is one of the most successful and long lasting biological control programs using entomopathogenic fungus in the world. However, studies to monitor the fate of this fungus on the sugarcane agroecosystem are rare, especially with respect to its persistence, efficacy in pest control and impact on the local populations of Metarhizium. The present study aimed at documenting the efficacy and persistence of M. anisopliae strain ESALQ1604 in a sugarcane field by using microsatellite molecular markers. The species diversity of Metarhizium was characterized in insects, soil and sugarcane roots in a sprayed and an unsprayed plot. Although the infection rates were not very high (≤ 50%), the applied strain was recovered from spittlebugs after 7, 30 and 60 days’ post-application, but accounted for only 50%, 50% and 70.5% of all insects killed by M. anisopliae, respectively. All haplotypes from spittlebug were associated with a single subclade of M. anisopliae. The highest haplotype diversity was found in soil (h = 0.989) and in the smallest in spittlebug (h = 0.779). Metarhizium robertsii, M. anisopliae, M. brunneum; one taxonomically unassigned lineage was found in soil and only M. brunneum and M. anisopliae were isolated from roots. This study revealed the great diversity of Metarhizium spp. in the sugarcane agroecosystem and the importance of the local population of M. anisopliae on spittlebugs management.

The influence of the temperature of aqueous conidial sprays on conidial viability and virulence against Diatraea saccharalis was evaluated for pure conidia, rice + fungus (technical concentrates) and oil-based formulations of Beauveria bassiana s.s. and Metarhizium anisopliae s.s. under laboratory conditions. The fungal preparations were suspended in water and maintained at 26 °C, 36 °C and 46 °C for one, four and six hours. Conidial viability was determined by plating aliquots of each suspension onto PDA medium followed by incubation for 20-22 h and observing for viable conidia (germ tubes longer than diameter of conidia). Fungal virulence was determined by spraying suspensions onto third-instar larvae of D. saccharalis. In general, germination and virulence, particularly for unformulated conidia, were negatively affected by increases in water temperature and exposure time in suspension. However, the decrease in conidial viability in the oil-in-water emulsion was less than 7% for both species after 6 h of exposure at 36 °C, in contrast to reductions of 7-21% and 28-60% for the oil-free suspensions of B. bassiana and M. anisopliae, respectively. For the sprays of conidia in an oil-in-water emulsion previously exposed to elevated water temperatures for longer periods, the levels of insect mortality were higher than those of pure conidia or technical concentrates under identical conditions. Our results indicate that emulsifiable oil-based formulations can protect the conidia of both species of fungi from the adverse effects of high water temperatures before spraying in the field.

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