In Brazil, countless insect species attack and damage sugarcane (Saccharum officinarum L.), which is an extremely important crop since it is planted on more than 10 million hectares. Among these insects, the sugarcane billbug, Sphenophorus levis (Coleoptera: Curculionidae), is of great importance as the larvae open tunnels in the rhizome of the plant, causing high damage and losses. This insect is attracted mainly to vinasse, which is the liquid fraction generated from the alcohol production and discarded onto the sugarcane fields for fertigation. Toward a novel control method for S. levis, the native entomopathogenic nematode Steinernema rarum (Pam 25) was compared with S. carpocapsae (IL 1) and Heterorhabditis bacteriophora (HBEN01) in respect to their ability to search for larvae of two insect hosts (Galleria mellonella and S. levis) within the cane rhizome. The selected nematode S. rarum was also assessed for rate effects, its survival in vinasse and field efficacy to control sugarcane pests S. levis, Hyponeuma taltula (Lepidoptera, Erebidae) and Leucothyreus alvarengai (Coleoptera: Melolonthidae). Steinernema rarum exhibited superior virulence to G. mellonella and S. levis larvae inserted into the cane rhizomes (75–78% mortality) compared to S. carpocapsae (30–53%) and H. bacteriophora (18–28%). Vinasse affected S. rarum when infective juveniles were suspended in the liquid compost for more than 6 h but did not affect the nematode when kept on the straw and soil treated with the compost. Steinernema rarum tested at 1–3 × 108 infective juveniles/ha on the sugarcane field caused 74.1, 56.3 and 50.6% control of S. levis, L. alvarengai and H. taltula, respectively.

ABSTRACT:Bacillus has been widely studied and used for the control of pests and diseases. The adapted protocol proposed by POLANCZYK (2004) proved to be more efficient than the one by the World Health Organization (WHO, 1985) to isolate edaphic strains of Bacillus. However, it has not been assessed for isolation of endophytic strains, which are much less abundant in the nature and more difficult to be isolated. This study aimed to compare two methodological procedures for isolation of Bacillus, established by the WHO (1985) and by POLANCZYK (2004), regarding their efficiency for isolation of endophytics and edaphics Bacillus strains from inside the root tissue of sugarcane, as well as from the associated soil sample, collected from 11 locations; and to compare the density of bacteria in both environments. Endophytic and edaphic strains of Bacillus were isolated by both procedures. However, the isolation protocol performed by POLANCZYK (2004) made more efficient by having a greater number of colony forming units (CFU) per gram of soil and root indicating that this procedure is more useful, especially for isolation of endophytic strains of Bacillus, which are much less abundant in the nature than edaphic strains, being therefore more difficult to be isolated. Using the Polanczyk protocol (2004), Bacillus strains were recovered from all roots (endophytic) and soil (edaphic) samples of all the 11 fields, suggesting that the plant root may be another important source for isolation of Bacillus besides the soil. Higher densities of Bacillus were isolated from the edaphic environment compared with the endophytic environment, with significant differences when isolated by Polanczyk method (2004 (2004), quanto a sua eficiência para o isolamento de estirpes endofitas e edáficas de Bacillus originárias do interior do tecido radicular de cana-de-açúcar, bem como de amostras de solos associados, coletada de 11 locais; e comparar a densidade de bactérias em ambos os ambientes. As cepas endofíticas e edáficas de Bacillus foram isoladas por ambos os procedimentos. No entanto, o protocolo de isolamento realizado por POLANCZYK (2004) demonstrou-se mais eficiente por gerar maior número de unidades de formação de colônias (CFU) por grama de solo e raiz, indicando que esse procedimento é mais útil, especialmente para isolamento de estirpes endofíticas de Bacillus, que são muito menos abundantes na natureza do que as cepas edáficas, sendo, portanto, mais difíceis de serem isoladas. Usando o protocolo de POLANCZYK (2004), as cepas de Bacillus foram isoladas de todas as amostras de raízes (endofíticas) e de solo (edáficas) de todos os 11 campos, sugerindo que a raiz da planta pode ser outra fonte importante de isolamento de Bacillus além do solo. As densidades mais altas de Bacillus foram isoladas do ambiente edáfico em comparação com o ambiente endofítico, com diferenças significativas quando isoladas pelo método de POLANCZYK (2004). PALAVRAS-CHAVE: controle biológico; bactéria benéfica;solo; raiz; endósporos.