José Eduardo Marcondes de Almeida

Effect of Thiamethoxam on Entomopathogenic Microorganisms

The compatibility of entomopathogenic microorganisms with thiamethoxam and other insecticides was studied in vitro and under field conditions. The microorganisms tested were: Aschersonia aleyrodis, Bacillus thuringiensis, Baculovirus anticarsia (NPVAg), Beauveria bassiana, Hirsutella thompsonii, Metarhizium anisopliae, Nomuraea rileyi, Paecilomyces farinosus, Sporothrix insectorum and Verticillium lecanii. Two concentrations of each product were tested in the laboratory, based on the maximum and minimum recommended rates for application in the field. The products were added to specific culture medium for growth of the entomopathogens. Reproductive and vegetative growth were evaluated for fungi, and colony forming units (CFU) were evaluated for bacteria. For the field test, CFUs were considered for both fungi and bacteria and caterpillar mortality for the NPV of Anticarsia gemmatalis (Hueb.). Results showed that: (1) the action of the pesticides on the vegetative growth and sporulation of the microorganisms varied as a function of the chemical nature of the products, its concentration and the microbial species; (2) thiamethoxam was compatible with all microorganisms studied; (3) endosulfan, monocrotophos and deltamethrin were the insecticides that most affected B. thuringiensis, B. bassiana, M. anisopliae and S. insectorum; (4) thiamethoxam did not affect the inoculum potential of B. thuringiensis, B. bassiana or M. anisopliae when applied to bean crop (Phaseolus vulgaris); and (5) thiamethoxam did not affect the efficiency of the nuclear polyhedral virus of A. gemmatalis.

Seleção de isolados de Beauveria bassiana (Bals.) Vuill. e Metarhizium anisopliae (Metsch.) Sorok. contra o cascudinho Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae)

This study was carried out to evaluate entomopathogenic fungi isolates as microbial control agents of the lesser mealworm, Alphitobius diaperinus (Panzer). Larvae and adults were inoculated with conidial suspension of 99 isolates of Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae (Metsch.) Sorok. (105 to 109 conidia/ml). Vegetative growth on culture media and sporulation on culture media, cooked rice and lesser mealworm cadavers were also evaluated. Isolates of B. bassiana were more effective than the M. anisopliae isolates and larvae were more susceptible than adults. The isolates UNIOESTE 04 and UNIOESTE 02 were originally obtained from adults of the lesser mealworm and were the most efficient isolates. In addition, UNIOESTE 04 showed high level of sporulation on different culture media and is considered a potential useful tool to lesser mealworm control.

Seleção de isolados de Metarhizium anisopliae (Metsch.) sorok. contra a cigarrinha da raiz da cana-de-açúcar Mahanarva fimbriolata (Stål) (Hemiptera: Cercopidae) em laboratório

O corte mecanizado da cana-de-acucar resulta num acumulo da palhada depositada no solo, proporcionando temperatura e umidade favoraveis a cigarrinha-da-raiz, Mahanarva fimbriolata (Stal), que provoca serios problemas devido a sua ocorrencia generalizada e altas populacoes. A patogenicidade de 79 isolados do fungo Metarhizium anisopliae (Metsch.) Sorok, provenientes de diferentes hospedeiros e regioes do Pais, foi testada em laboratorio. Dez ninfas da cigarrinha foram colocadas em placas de Petri (9 cm de diâmetro) contendo folhas de cana lavadas e pulverizadas com 1 ml de uma suspensao contendo 5 x 107 conidios/ml. Cada tratamento foi composto por cinco repeticoes. Apos a pulverizacao, as placas foram colocadas em câmara climatizada a 25 ± 1oC, 70 ± 10% de UR e fotofase de 12h. A mortalidade foi avaliada diariamente, calculando-se os indices de mortalidade acumulada total, confirmada e corrigida de cada tratamento. Os melhores isolados testados foram: IBCB 348, IBCB 351, IBCB 363, IBCB 408, IBCB 410, IBCB 418, IBCB 425 e IBCB 482, os quais apresentaram mortalidade confirmada igual ou superior a 70% seis dias apos a pulverizacao.

Controle do cupim subterrâneo Heterotermes tenuis (Hagen) com iscas termitrap impregnadas com inseticidas e associadas ao fungo entomopatogênico Beauveria bassiana (Bals.) Vuill.

The control of the termite Heterotermes tenuis (Hagen) (Isoptera: Rhinotermitidae) was evaluated using the bait/trap Termitrap impregnated with insecticides in low concentrations and associated to the entomopathogenic fungus Beauveria bassiana (Bals.) Vuill. isolate 634 (from Solenopsis invicta F. Smith) in sugarcane (Saccharum officinarum L.). The experiment was conducted in Rio Claro-SP, in a sugarcane field highly infested with termites. The treatments consisted of: imidacloprid 0,01%; imidacloprid 0,01% + B. bassiana; WG 0,003%; WG 0,003% + B. bassiana; B. bassiana; and Untreated control. Each treatment was replicated five times. The insecticides were impregnated on baits by immersion in water, being their concentrations calculated according to the weigth of the bait, and the B. bassiana was applied as pure conidia (109 conidia/bait). The research was conducted during November 1996 through April 1997. The evaluations were made after 15, 30, 41, 63, 86 e 136 days by assigning indexes to populations levels. All treatments significantly reduced termite populations when compared to the control. However, it took longer for B. bassiana alone to reduced H. tenuis population. The treatments with imidacloprid and WG were the most efficient in the control of termites in sugarcane. The baits/traps did not repel the termites.

Patogenicidade de Beauveria bassiana ao psilídeo Diaphorina citri e compatibilidade do fungo com produtos fitossanitários

The objective of this work was to evaluate the pathogenicity of Beauveria bassiana to Diaphorina citri (Hemiptera: Psyllidae) nymphs, and to check the compatibility of the fungus with phytosanitary products, and its persistence in citrus plants. Nymphs of D. citri were sprayed with B. bassiana in the concentrations 5x106, 1x107, 5x107, 1x108, 5x108 and 1x109 conidia mL-1 for lethal concentration determination. In order to evaluate the compatibility of the phytosanitary products with the fungus, neem extract and five insecticides of four different chemical groups were individually added to PDA culture medium, in which the fungus was cultivated. Vegetative growth, sporulation and viability of the entomopathogen were evaluated. In the greenhouse, citrus plants were first treated with the phytosanitary products and, then, with the pathogen. Exposure times of 24 hours, 7 and 14 days were evaluated. Fungus pathogenicity to D. citri nymphs was confirmed; LC50 of 0.4 x107 and LC90 of 6.7 x107 conidia mL-1 were determined at the tenth day of evaluation. In the laboratory, the phytosanitary products reduced fungus growth. In greenhouse, the products did not affect fungus survival on citrus plants.

FORAGING AREA OF THE UNDERGROUND TERMITE Heterotermes tenuis (ISOPTERA; RHINOTERMITIDAE) IN SUGARCANE

The area of foraging of the termite species Heterotermes tenuis was evaluated using the Termitrap® bait marked with the radioisotope 32P. The experiment was conducted in an area with sugarcane, in Piracicaba,SP Brazil. Twenty infestion foci of H. tenuis were identified with baits. Each focus recieved baits labeled with 18.5 MBb (500 mCi) 32P in the form of sodium phosphate diluted in 15 ml of distilled water. Surrounding the marked baits were installed baits without marker in four points: North, South, East and West, halfway of 1, 5, 10 and 20 meters. Each treatment was represented previously by a focus settled and marked and one of the distances specified for the monitoring bait, being repeated of 4 to 6 times. After 15 days of the demarcation, the evaluation was proceeded by collecting 15 individuals of H. tenuis from each infested monitoring bait. These insects were transferred to glass flasks with distilled water, and analyzed in a liquid cintilador, by the test of Cerenkov. It was verified that in only 20% of the monitoring baits termites were collected and 11.3% were marked with the radioisotope, and that several colonies may by foraging in a same area. The foraging area for a colony of H. tenuis in sugarcane can vary from 3 to 1,250 m2.

Eficiência de isolados de Metarhizium anisopliae (Metsch.) Sorok. no controle da cigarrinha-da-raiz da cana-de-açúcar, Mahanarva fimbriolata (Stal, 1854) (Hemiptera: Cercopidae), em condições de campo

No Estado de Sao Paulo, Mahanarva fimbriolata (Stal) e observada a partir de setembro e, atual-mente, o controle esta sendo realizado com a aplicacao do fungo Metarhizium anisopliae (Metsch.) Sorok. O objetivo deste trabalho foi comparar a eficiencia de isolados de M. anisopliae em controlar populacoes naturais de M. fimbriolata em cultivos comerciais de cana-de-acucar colhida mecanicamente. Os isolados IBCB 348, IBCB 408, IBCB 410 e IBCB 425 do fungo foram aplicados na concentracao de 1,5 × 1012 conidios/ha, utilizando-se 2 kg de arroz + fungo. As suspensoes de conidios foram preparadas pela lavagem do arroz+fungo em agua. O delineamento experimental foi em blocos casualizados com 5 tratamentos e 4 repeticoes, sendo cada repeticao composta por 7 linhas de 100 m de comprimento e espacamento de 1,5 m. Decorridos 30 dias da pulverizacao, observaram-se que os isolados IBCB 408 e IBCB 425 apresentaram eficiencia de controle de ninfas de 63 e 62%, respectivamente, e para os adultos a eficiencia foi de 100%. Aos 60 dias da aplicacao, o isolado IBCB 425 foi o mais eficiente para controlar as ninfas (48,4%). De maneira geral, nao ocorreu diferenca estatistica entre os isolados quanto a capacidade de controle das populacoes de M. fimbriolata. Observou-se que apenas uma unica aplicacao de M. anisopliae nao e eficiente para a manutencao da populacao de M. fimbriolata abaixo do nivel de dano economico.

Produção líquida de fungos entomopatogênicos e efeitos da radiação ultravioleta e da temperatura sobre os propágulos produzidos

The purpose of this paper was to evaluate the liquid culture media for the production of Metarhizium anisopliae (IBCB 425) and Beauveria bassiana (IBCB 66), as well as the tolerance of these seedlings to the ultraviolet action and to the temperature. Twelve treatments composed of combinations between carbon and nitrogen concentrations were assessed. In order to determine the effect of ultraviolet radiation, plates with blastos-pores were exposed to it for 25 and 50 seconds. To determine the temperature efect, blastospores from culture media were exposed to 20, 25, 30 and 35°. For the virulence experiments, caterpillars of Diatraea saccharalis were sprayed with 2 mL of fungal suspension with the aid of a Potter tower. Te best media for M. anisopliae are 16.00 g (carbon) + 7.00 g (nitrogen) and 14.40 g (carbon) + 7.00 g (nitrogen), whereas for B. bassiana: 20.00 g (carbon) + 6.30 g (nitrogen) and 20.00 g (carbon) + 7.00 g (nitrogen). Te longer the exposure to ultraviolet radiation, the smaller the number of colonies. At 35°, there is a significant decrease in the formation of colonies. Te produced seedlings of fungi are pathogenic to D. saccharalis.

Seleção de isolados de fungos entomopatogênicos para o controle de Leptopharsa heveae (Hemiptera: Heteroptera, Tingidae)

Twenty-seven strains of entomopathogenic fungi from diverse hosts and geographic regions were tested for their virulence, total conidia production and viability rate. The conidia production was made in PDA medium. Leptopharsa heveae third instar nymphs were inoculated with fungi suspension at the concentration of 1 x 109 conidia/mL. In each two days, confirmed mortality evaluations were done and a large variation of virulence was observed, been 1,189, of Metarhizium anisopliae, the best one. The top five strains, plus the two standards, were then submitted to production tests. For this purpose, pre-matrices in PDA medium were prepared, from which matrices in rice medium were after inoculated at the concentration of 5 x 107 conidia/mL. From the rice matrices, polypropylene bags with maize medium were inoculated with 40 mL of these suspensions. After fifteen days of incubation, the bags content were evaluated for total conidia production and their viability. The 1200 strain, of Isaria fumosorosea, presented the best results in total yield and viability rate; to E9 strain of de Metarhizium anisopliae was more virulent to rubber tree lacebug.