Endophytic and entomopathogenic strains of Beauveria sp to control the bovine tick Rhipicephalus (Boophilus) microplus

Pathogenicity of strains of the entomopathogenic fungus Beauveria bassiana and endophytic strains of Beauveria sp against the bovine tick Rhipicephalus (Boophilus) microplus was tested in laboratory bioassays and under field conditions. Suspensions containing 10(5), 10(7) and 10(9) conidia/mL were prepared of each fungal strain for laboratory bioassays. The ticks were maintained at 28 degrees C, 90 +/- 5% relative humidity, and the following variables were evaluated: initial female weight, egg weight, hatching percentage, reproductive efficiency, and percentage control. For tests under field conditions, a Beauveria suspension containing 10(6) conidia/mL was sprayed on tick-infested cows. After 72 h, the ticks were collected to estimate mortality under field conditions. Laboratory bioassays showed a mortality of 20 to 50% of the ticks seven days after inoculation with 10(7) Beauveria conidia/mL. Under field conditions 10(6) Beauveria conidia/mL induced 18-32% mortality. All Beauveria strains were effective in biological control of R. (Boophilus) microplus under laboratory and field test conditions. This is the first demonstration that endophytic fungi can be used for biological control of the cattle tick; this could help reduce environmental contamination by diminishing the need for chemical acaricides. Two endophytic strains were isolated from maize leaves and characterized by molecular sequencing of 5.8S rDNA ITS1 and ITS2 and morphological analyses of conidia. We found that these two endophytic Beauveria isolates, designated B95 and B157, are close to Beauveria amorpha.

A multi-screening approach for marine-derived fungal metabolites and the isolation of cyclodepsipeptides from Beauveria felina

Extracts obtained from 57 marine-derived fungal strains were analyzed by HPLC-PDA, TLC and ¹H NMR. The analyses showed that the growth conditions affected the chemical profile of crude extracts. Furthermore, the majority of fungal strains which produced either bioactive of chemically distinctive crude extracts have been isolated from sediments or marine algae. The chemical investigation of the antimycobacterial and cytotoxic crude extract obtained from two strains of the fungus Beauveria felina have yielded cyclodepsipeptides related to destruxins. The present approach constitutes a valuable tool for the selection of fungal strains that produce chemically interesting or biologically active secondary metabolites.

The effect of mode of exposure to Beauveria bassiana on conidia acquisition and host mortality of Colorado potato beetle, Leptinotarsa decemlineata

The effects of the mode of exposure of second instar Colorado potato beetles to Beauveria bassiana on conidia acquisition and resulting mortality were investigated in laboratory studies. Larvae sprayed directly with a B, bassiana condial suspension, larvae exposed to B, bassiana-treated foliage, and larvae both sprayed and exposed to treated foliage experienced 76, 34, and 77% mortality, respectively. The total number of conidia and the proportion of germinating conidia were measured over time for four sections of the insect body: the ventral surface of the head (consisting mostly of ventral mouth parts), the ventral abdominal surface, the dorsal abdominal surface, and the legs. From observations at 24 and 36 h posttreatment, mean totals of 161.1 conidia per insect were found on sprayed larvae, 256.1 conidia on larvae exposed only to treated foliage, and 408.3 conidia on larvae both sprayed and exposed to treated foliage, On sprayed larvae, the majority of conidia were found on the dorsal abdominal surface, whereas conidia were predominantly found in the ventral abdominal surface and mouth parts on larvae exposed to treated foliage, Between 24 and 36 h postinoculation the percentage of conidia germinating on sprayed larvae increased slightly from 80 to 84%), On the treated foliage, the percentage of germinated conidia on larvae increased from 35% at 24 h to 50% at 36 h posttreatment, Conidia germination on sprayed larvae on treated foliage was 65% at 24 h and 75% at 36 h posttreatment, It is likely that the gradual acquisition of conidia derived from the continuous exposure to B. bassiana inoculum on the foliar surface was responsible for the increase in germination over time on larvae exposed to treated foliage, The density and germination of conidia were observed 0, 4, 8, 12, 16, 20, and 24 h after being sprayed with or dipped in conidia suspensions or exposing insects to contaminated foliage, Conidia germinated twice as fast on sprayed insects as with any other treatment within the first 12 h, This faster germination may be due to the pressure of the sprayer enhancing conidial lodging on cuticular surfaces. (C) 2001 Academic Press.

Evaluation of synergistic interactions between the Colorado potato beetle (Coleoptera : Chrysomelidae) pathogen Beauveria bassiana and the insecticides, imidacloprid, and cyromazine

Laboratory studies investigated the interaction between the fungal entomopathogen Beauveria bassiana (Balsamo) Vuillemin and sublethal doses of the insecticides imidacloprid and cyromazine when applied to larvae of the Colorado potato beetle, Leptinotarsa decemlinenta (Say). When second instars were fed potato leaf discs treated with sublethal doses of imidacloprid and a range of doses of B. bassiana, a synergistic action was demonstrated. Similar results were observed when larvae were sprayed directly with B. bassiana conidia and immediately fed leaf discs treated with imidacloprid. No synergistic interaction was detected when larvae were fed leaf discs treated with sublethal doses of imidacloprid 24 h after application of R. bassiana conidia to larvae. However, a synergistic interaction was detected when larvae were fed leaf discs treated with imidacloprid and sprayed with B, bassiana conidia 24 h later. Although sublethal doses of both imidacloprid and the triazine insect growth regulator (IGR) cyromazine prolonged the duration of the second instar, only imidacloprid interacted with B. bassiana to produce a synergistic response in larval mortality. In leaf consumption studies, the highest dose of B, bassiana tested promoted feeding in inoculated second instars. Feeding was inhibited when larvae were fed foliage treated with sublethal doses of imidacloprid and significantly reduced when fed foliage treated with a sublethal dose of cyromazine. Starvation of larvae for 24 h immediately after B. bassiana treatment produced a similar result to the combined treatment of B. bassiana and imidacloprid and increased the level of mycosis when compared with B. bassiana controls. Imidacloprid treatment affected neither the rate of germination of B. bassiana conidia on the insect cuticle nor the rate at which conidia were removed from the integument after application. The statistical analysis used to detect synergism and the possible role of starvation-induced stress factors underlying the observed synergistic interactions are discussed.

Infecção de Conotrachelus humeropictus Fiedler (Coleoptera: Curculionidae) por Metarhizium anisopliae (Metsch.) Sor. e Beauveria bassiana (Bals.) Vuill. no solo

Foi avaliada a infecção de larvas de Conotrachelus humeropictus Fiedler, séria praga do cacaueiro (Theobroma cacao L.) e do cupuaçuzeiro (T. grandiflorum (Willd. ex Spreng.) Schum.) na Amazônia brasileira, por Metarhizium anisopliae (Metsch.) Sor. e Beauveria bassiana (Bals.) Vuill. A pesquisa foi desenvolvida nos campos experimentais da Comissão Executiva do Plano da Lavoura Cacaueira - CEPLAC, em Ouro Preto D'Oeste, Rondônia. Foram testadas suspensões de 3,93 χ 1010 conídios/ml de M. anisopliae e 4,26 χ 1010 conídios/ml de B. bassiana, pulverizadas superficialmente em solo contido em recipientes de PVC, onde em diferentes dias após a pulverização (um, três, sete e quatorze dias) liberou-se larvas do último ínstar da praga. Beauveria bassiana mostrou-se mais eficiente (52,0% de mortalidade) do que M. anisopliae (42,7%), evidenciando assim, seu maior potencial no controle da praga. Os índices de mortalidade foram estatisticamente iguais para larvas liberadas até o 7º dia da contaminação, decrescendo significativamente no 14º dia. A queda na efetividade pode estar associada à presença de microrganismos antagonistas no solo.

Ocorrência de Beauveria bassiana (Bals.) Vuill. em broca-do-café (Hypothenemus hampei, ferrari) no estado de Rondônia, Brasil1

Em trabalho efetuado visando o levantamento de inimigos naturais da broca-do-café (Hypothenemus hampei, Ferrari) no estado de Rondônia, verificou-se a ocorrência do fungo Beauveria bassiana (Bals.) Vuill. conforme identificação da Embrapa Rondônia. Os frutos de café foram coletados em cafezais da cultivar Conilon (Coffea canephora), nos municípios de Rolim de Moura, Ouro Preto do Oeste e Machadinho do Oeste, Rondônia, nos meses de fevereiro a março de 2000. A percentagem média de infecção de B. bassiana em frutos broqueados variou de 0,23% a 0,47%, sendo menor que em outras regiões do Brasil.

Relative susceptibility of different stages of Rhodinius Prolixus to the entomopathogenic Hyphomycete Beauveria Bassiana

Laboratory bioassays were conducted to determine the relative suscepbility of eggs, 1st-, 3rd-, 5th- instar nymphs and adults of Rhodnius prolixus to one isolate of the entomopathogenic hyphomycete, Beauveria bassiana. Treatments consisted of directly spraying on insects of increasing doses of inoculum (3 x 10* to 3 x 10 (elevated to 5th potency) conidia per cm*). Mortality due to all doses of conidia was very high in the five tested stages of the target insect. Experiments on eggs demonstrated that the fungal isolate was able to kill eggs before they hatched. Both time-mortality and dose-mortality responses showed that the susceptibility of R. prolixus varied according to its stage of development and increased with age. As matter of fact, at the dose of 3 x 10* conidia per cm*, LD50 varied between 11.2 days in 1st-instar nymphs and 6.4 days in both 5th-instar nymphs and adults. Comparison of LD50 permitted to estimate that 1st-instar nymphs were about 700-fold less susceptible than the two oldest stages

Selection of Beauveria bassiana and Metarhizium anisopliae Isolates to Control Triatoma infestans

Twenty three isolates of Beauveria bassiana and 13 isolates of Metarhizium anisopliae were tested on third instar nymphs of Triatoma infestans, a serious vector of Chagas disease. Pathogenicity tests at saturated humidity showed that this insect is very susceptible to fungal infection. At lower relative humidity (50%), conditions expected in the vector microhabitat, virulence was significantly different among isolates. Cumulative mortality 15 days after treatment varied from 17.5 to 97.5%, and estimates of 50% survival time varied from 6 to 11 days. Maintaining lower relative humidity, four B. bassiana and two M. anisopliae isolates were selected for analysis of virulence at different conidial concentrations and temperatures. Lethal concentrations sufficient to kill 50% of insects (LC50) varied from 7.1x105 to 4.3x106 conidia/ml, for a B. bassiana isolate (CG 14) and a M. anisopliae isolate (CG 491) respectively. Most isolates, particularly B. bassiana isolates CG 24 and CG 306, proved to be more virulent at 25 and 30°C, compared to 15 and 20°C. The differential virulence at 50% humidity observed among some B. bassiana isolates was not correlated to phenetic groups in cluster analysis of RAPD markers. In fact, the B. bassiana isolates analyzed presented a high homogeneity (> 73% similarity).

Activity of oil-formulated Beauveria bassiana against Triatoma sordida in peridomestic areas in central Brazil

Field tests were carried out during the rainy season of 2001/2002 in São Luís de Montes Belos, Goiás, Brazil, to evaluate the potential of the entomopathogenic fungus, Beauveria bassiana, against peridomestic Triatoma sordida. An oil-water formulation of the isolate CG 14 (Embrapa) was applied in triatomine infested hen houses of four farms at a final concentration of 10(6) conidia/cm². Numbers of T. sordida decreased over the next 25 days, after application of the fungus, and B. bassiana developed on dead insects in one hen house. A high number of B. bassiana colonies was detected in substrates collected in treated hen houses 24 h after application of CG 14. In the following three months the presenceof B. bassiana declined to values found before treatment.

Patogenicidade de isolados de Metarhizium anisopliae (Metsch.) Sorokin e de Beauveria bassiana (Bals.) Vuillemin a Scaptocoris carvalhoi Becker (Hemiptera, Cydnidae)

Avaliou-se a patogenicidade de isolados dos fungos Metarhizium anisopliae (Metsch.) Sorokin e Beauveria bassiana (Bals.) Vuillemin a Scaptocoris carvalhoi Becker, 1967 sob condições de laboratório e em casa de vegetação. Os experimentos foram conduzidos na Embrapa Agropecuária Oeste de Dourados, MS, durante o ano de 2003. Foram avaliados dez isolados de M. anisopliae e onze de B. bassiana, em laboratório, utilizando-se o delineamento inteiramente casualizado, com cinco repetições (10 adultos e 5 ninfas/parcela). A patogenicidade de M. anisopliae (Ma69) também foi testada em ninfas e adultos, separadamente, em laboratório e casa de vegetação. Os níveis de mortalidade do percevejo foram maiores com os isolados de M. anisopliae que variaram de 73,3% a 94,7% contra 10,7% a 78,7% para os de B. bassiana. Em casa de vegetação, a porcentagem de mortalidade do percevejo causada pelo fungo Ma69 foi de 57,3% e não foi constatada diferença por este isolado quanto à mortalidade de ninfas e adultos, em laboratório. Todavia, em casa de vegetação, os níveis de mortalidade foram significativamente maiores (p<0,05) para ninfas (38,4%) do que para os adultos (16,2%). Com base nos resultados obtidos, concluiu-se que o isolado Ma69 de M. anisopliae foi mais eficiente para S. carvalhoi tanto em laboratório quanto em casa de vegetação, constituindo em uma alternativa promissora para ser utilizado como inseticida microbiano em condições de campo.

Seleção de isolados de Beauveria bassiana patogênicos ao bicudo-do-algodoeiro

Objetivou-se selecionar isolados de Beauveria bassiana (Balsamo) Vüillemin provenientes de diferentes hospedeiros e regiões geográficas, patogênicos ao Anthonomus grandis, o bicudo-do-algodoeiro. Foram analisados 12 isolados em condições de laboratório. Os isolados obtidos originalmente de A. grandis foram pouco virulentos a essa praga. A mortalidade do bicudo teve início no segundo dia após a inoculação das suspensões fúngicas, variando de 15% a 83%, com TL50 entre 5,30 a 11,06 dias. Os isolados apresentaram variabilidade quanto à germinação dos conídios em meio de cultura artificial, e esta não se correlacionou com a patogenicidade. O isolado CG138 de B. bassiana destacou-se como um dos mais virulentos ao bicudo-do-algodoeiro.

Compatibilidade entre Beauveria bassiana e o predador Chrysoperla externa em laboratório

O objetivo deste trabalho foi avaliar os efeitos do fungo Beauveria bassiana sobre ovos e larvas de Chrysoperla externa. Ovos com até 24 horas de idade e insetos no 1º, 2º e 3º ínstares foram imersos em suspensões fúngicas de 1,0x10(4) a 1,0x10(8) conídios mL-1. Não houve efeito do fungo sobre a viabilidade dos ovos. Larvas de terceiro ínstar foram afetadas por B. bassiana, e as suspensões de 1,0x10(7) e 1,0x10(8) conídios mL-1 interferiram nos parâmetros avaliados.

Controle associado de Alphitobius diaperinuse efeito de microrganismos eficazes no desenvolvimento de Beauveria bassiana

O objetivo deste trabalho foi avaliar a eficiência de Beauveria bassiana (Bb), terra diatomácea (TD) e microrganismos eficazes (EM-4), associados ou não, no controle de Alphitobius diaperinus, e o efeito de EM-4 no desenvolvimento de B. bassiana. Os agentes de controle (Bb, TD e EM-4), em diferentes concentrações e combinações, foram aplicados em uma mistura de cama-de-frango e ração, em que os insetos foram acondicionados por dez dias. Avaliaram-se, in vitro: a germinação, as unidades formadoras de colônia (UFC), o crescimento vegetativo e a produtividade de conídios de Bb em contato com suspensão aquosa de EM-4 (1%) não filtrada e filtrada. Os maiores índices de mortalidade foram observados nos tratamentos TD + Bb + EM-4 e TD + Bb, nas maiores concentrações. Verificou-se efeito não-aditivo sinérgico para TD + Bb nas três concentrações. Nos testes com EM-4 filtrado, não houve diferença em relação à testemunha quanto à germinação e às UFC, entretanto, o crescimento vegetativo e a produtividade de conídios foram negativamente afetados. O uso conjunto de B. bassiana e terra diatomácea, para o manejo de populações de A. diaperinus, pode reduzir o uso de produtos químicos.

Molecular characterization and pathogenicity of isolates of Beauveria spp. to fall armyworm

The objective of this work was to evaluate the pathogenicity of 24 Beauveria isolates to Spodoptera frugiperda larvae, and characterize them molecularly through rDNA-ITS sequencing and RAPD markers. Sequencing of rDNA-ITS fragments of 570 bp allowed the identification of isolates as B. bassiana or B. brongniarti by sequence comparison to GenBank. Sixty seven polymorphic RAPD fragments were capable to differentiate 20 among 24 Beauveria isolates, grouping them according to the derived host insect and to pathogenicity against maize fall armyworm larvae. Three RAPD markers were highly associated to the pathogenicity against S. frugiperda, explaining up to 67% of the phenotypic variation. Besides identification and molecular characterization of Beauveria isolates, ITS sequence and RAPD markers proved to be very useful in selecting the isolates potentially effective against S. frugiperda larvae and in monitoring field release of these microorganisms in biocontrol programs.