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Sample records for fungi metarhizium anisopliae

  1. Anopheline and culicine mosquitoes are not repelled by surfaces treated with the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana

    NARCIS (Netherlands)

    Mnyone, L.L.; Koenraadt, C.J.M.; Lyimo, I.N.; Mpingwa, M.W.; Takken, W.; Russell, T.L.

    2010-01-01

    Background - Entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana, are promising bio-pesticides for application against adult malaria mosquito vectors. An understanding of the behavioural responses of mosquitoes towards these fungi is necessary to guide development of fungi beyond t

  2. Genome sequencing and comparative transcriptomics of the model entomopathogenic fungi Metarhizium anisopliae and M. acridum.

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    Gao, Qiang; Jin, Kai; Ying, Sheng-Hua; Zhang, Yongjun; Xiao, Guohua; Shang, Yanfang; Duan, Zhibing; Hu, Xiao; Xie, Xue-Qin; Zhou, Gang; Peng, Guoxiong; Luo, Zhibing; Huang, Wei; Wang, Bing; Fang, Weiguo; Wang, Sibao; Zhong, Yi; Ma, Li-Jun; St Leger, Raymond J; Zhao, Guo-Ping; Pei, Yan; Feng, Ming-Guang; Xia, Yuxian; Wang, Chengshu

    2011-01-06

    Metarhizium spp. are being used as environmentally friendly alternatives to chemical insecticides, as model systems for studying insect-fungus interactions, and as a resource of genes for biotechnology. We present a comparative analysis of the genome sequences of the broad-spectrum insect pathogen Metarhizium anisopliae and the acridid-specific M. acridum. Whole-genome analyses indicate that the genome structures of these two species are highly syntenic and suggest that the genus Metarhizium evolved from plant endophytes or pathogens. Both M. anisopliae and M. acridum have a strikingly larger proportion of genes encoding secreted proteins than other fungi, while ~30% of these have no functionally characterized homologs, suggesting hitherto unsuspected interactions between fungal pathogens and insects. The analysis of transposase genes provided evidence of repeat-induced point mutations occurring in M. acridum but not in M. anisopliae. With the help of pathogen-host interaction gene database, ~16% of Metarhizium genes were identified that are similar to experimentally verified genes involved in pathogenicity in other fungi, particularly plant pathogens. However, relative to M. acridum, M. anisopliae has evolved with many expanded gene families of proteases, chitinases, cytochrome P450s, polyketide synthases, and nonribosomal peptide synthetases for cuticle-degradation, detoxification, and toxin biosynthesis that may facilitate its ability to adapt to heterogeneous environments. Transcriptional analysis of both fungi during early infection processes provided further insights into the genes and pathways involved in infectivity and specificity. Of particular note, M. acridum transcribed distinct G-protein coupled receptors on cuticles from locusts (the natural hosts) and cockroaches, whereas M. anisopliae transcribed the same receptor on both hosts. This study will facilitate the identification of virulence genes and the development of improved biocontrol strains with

  3. Differential expression of genes involved in entomopathogenicity of the fungi Metarhizium anisopliae var. anisopliae and M. anisopliae var. acridum (Clavicipitaceae).

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    Carneiro-Leão, M P; Andreote, F D; Araújo, W L; Oliveira, N T

    2011-05-03

    Expression analysis of the genes involved in germination, conidiogenisis and pathogenesis of Metarhizium anisopliae during its saprophytic and pathogenic life stages can help plan strategies to increase its efficacy as a biological control agent. We quantified relative expression levels of the nitrogen response regulator gene (nrr1) and a G-protein regulator of genes involved in conidiogenesis (cag8), using an RT-qPCR assay. Comparisons were made between M. anisopliae var. anisopliae and M. anisopliae var. acridum during germination and conidiogenesis and at different stages of pathogenesis. The cag8 gene was repressed during germination and induced during conidial development and the pathogenic phase, and the nrr1 gene was induced during germination, conidiogenesis and the pathogenic phase. Both genes were more expressed in M. anisopliae var. anisopliae, demonstrating that different varieties of M. anisopliae differ in activation of genes linked to virulence for certain environments and hosts. This suggests that differences among these varieties in the ability to adapt could be attributed not only to specific genomic regions and genes, but also to differential gene expression in this fungus, modulating its ability to respond to environmental stimuli.

  4. In vivo interactions of entomopathogenic fungi, Beauveria spp. and Metarhizium anisopliae with selected opportunistic soil fungi of sugarcane ecosystem.

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    Geetha, N; Preseetha, M; Hari, K; Santhalakshmi, G; Bai, K Subadra

    2012-07-01

    In the present study, the interactions of entomopathogenic fungi viz., Beauveria bassiana, Beauveria brongniartii and Metarhizium anisopliae among themselves and three other opportunistic soil fungi from the sugarcane ecosystem namely, Fusarium saachari, Aspergillus sp. and Penecillium sp. were assayed in vivo against Galleria mellonella larvae. The tested fungi were co-applied on IV instar G. mellonella @ 1 x 10(7) ml(-1), in combinations of two, at the interval of 24 hrs either preceding or succeeding each otherto assess their efficacy and sporulation rates. Results showed that often mortality rates did not correspond to the spore harvest of the mortality agent and presence of other fungus may be antagonistic. The efficacy of B. bassiana (90%) and B. brongniartii (100%) was not enhanced further but was negatively affected in most combinations with other fungi. In case of M. anisopliae compatibility was higher, resulting in higher mortality by application of B. bassiana before (100%) or after (83.3%) M. anisopliae than when it was applied alone (70%). During sporulation, B. bassiana faced the most intense competition from M. anisopliae (2.75 x 10(6) larva(-1)) and enhancement due to F sacchari irrespective of sequence of application. In case of B. brongniartii, sporulation was lowest in the combination of B. brongniartiipreceding M. anisopliae (1.83 x10(6) larva(-1)) and B. brongniartii succeeding B. bassiana (1.58 x 10(6) larva(-1)). Of all fungi tested, except F sacchari (65.33 x 10(6) larva(-1)) all the other species affected sporulation of M. ansiopliae with the least in treatment of B. bassiana application following M. anisopliae. Similar kind of interaction was observed during sporulation of soil fungi when combined with entomopathogenic fungi, though individually they could not cause mortality of larvae.

  5. Virulence of Entomopathogenic Fungi Metarhizium anisopliae and Paecilomyces fumosoroseus for the Microbial Control of Spodoptera exigua.

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    Han, Ji Hee; Jin, Byung Rae; Kim, Jeong Jun; Lee, Sang Yeob

    2014-12-01

    The beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) is difficult to control using chemical insecticides because of the development of insecticide resistance. Several pest control agents are used to control the beet armyworm. Entomopathogenic fungi are one of the candidates for eco-friendly pest control instead of chemical control agents. In this study, among various entomopathogenic fungal strains isolated from soil two isolates were selected as high virulence pathogens against larva of beet armyworm. Control efficacy of fungal conidia was influenced by conidia concentration, temperature, and relative humidity (RH). The isolates Metarhizium anisopliae FT83 showed 100% cumulative mortality against second instar larvae of S. exigua 3 days after treatment at 1 × 10(7) conidia/mL and Paecilomyces fumosoroseus FG340 caused 100% mortality 6 days after treatment at 1 × 10(4) conidia/mL. Both M. anisopliae FT83 and P. fumosoroseus FG340 effectively controlled the moth at 20~30℃. M. anisopliae FT83 was significantly affected mortality by RH: mortality was 86.7% at 85% RH and 13.4% at 45% RH. P. fumosoroseus FG340 showed high mortality as 90% at 45% RH and 100% at 75% RH 6 days after conidia treatments. These results suggest that P. fumosoroseus FG340 and M. anisopliae FT83 have high potential to develop as a biocontrol agent against the beet armyworm.

  6. Optimization of compatible non-ionic surfactant for formulation development of hydrophobic conidia of entomopathogenic fungi, Beauveria bassiana (Hypocreales:Cordycipitaceae) and Metarhizium anisopliae Hypocreales:Clavicipita

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    Aerial conidia, especially dried conidia of entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae are hydrophobic, and therefore surfactants are needed for developing water-based formulations in laboratory studies, greenhouse bioassays, and field trials as well as commercial product ...

  7. Preservation of aerial conidia and biomasses from entomopathogenic fungi Beauveria brongniartii and Metarhizium anisopliae during lyophilization.

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    Toegel, Stefan; Salar-Behzadi, Sharareh; Horaczek-Clausen, Andrea; Viernstein, Helmut

    2010-09-01

    In this study, we assessed the stability provided by different formulations to aerial conidia or biomasses (conidia, blastospores, and mycelia) of Beauveria brongniartii and Metarhizium anisopliae subjected to lyophilization. First, the impact of the freezing and drying processes on spore survival was evaluated. Whereas unprotected B. brongniartii spores showed high cryosensitivity, those of M. anisopliae were markedly harmed by the drying process. Then, the protective efficiency of 14 excipients was systematically evaluated and optimized regarding required concentrations. Fructose, glucose, and saccharose significantly enhanced viabilities for B. brongniartii and M. anisopliae spores following lyophilization, especially as a result of their cryoprotective effects. In addition, the effect of various bulking agents on spore survival was studied and dextran 4 was selected to enhance the physical properties of the lyophilized products. The combination of fructose and dextran 4 was further applied to prepare lyophilized biomasses of both fungi. In comparison to freshly harvested biomasses, the lyophilized products showed similar growth rates and a comparable production of virulent secondary metabolites such as destruxin A, destruxin B, or oosporein, suggesting their applicability as biological control agents.

  8. Compatibility of polymers to fungi Beauveria bassiana and Metarhizium anisopliae and their formulated products stability

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    Inajá Marchizeli Wenzel Rodrigues

    2017-08-01

    Full Text Available Stability during storage is a limiting factor in the development and use of bioinsecticides. This study aims to evaluate the compatibility of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana with components used in encapsulated formulations and to evaluate shelf-life in formulations with B. bassiana. Sodium alginate and maltodextrin, hemicellulose, and dimethyl sulfoxide solvents were evaluated. Compatibility was evaluated by mixing the components with PDA, and the parameters evaluated were vegetative growth, sporulation and viability. The formulations of fungi were prepared in various concentrations of sodium alginate and then added drop-wise to calcium chloride. To evaluate the stability, formulations were stored at 26.5, 4.0, and -20.0°C. Monthly, samples were taken, and the capsules were placed on Petri dishes with PDA to verify the growth after seven days of incubation. These evaluations were performed by the 12th month. Biopolymers evaluated were compatible with both fungi, exception maltodextrin at 1.5%, which was classified as moderately toxic to B. bassiana. Dimethyl sulfoxide was classified as moderately toxic to B. bassiana at 3.0% and to M. anisopliae at 2.0% and 3.0%. The formulations were stable throughout the 12 months in the conditions evaluated, while for the pure conidia of the fungus, 46% viability in the 6th month at 26.5°C was observed.

  9. MALDI-TOF mass spectrometry applied to identifying species of insect-pathogenic fungi from the Metarhizium anisopliae complex.

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    Lopes, Rogério Biaggioni; Faria, Marcos; Souza, Daniela Aguiar; Bloch, Carlos; Silva, Luciano P; Humber, Richard A

    2014-01-01

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has proven to be a powerful tool for taxonomic resolution of microorganisms. In this proof-of-concept study, we assessed the effectiveness of this technique to track the current gene sequence-based phylogenetic classification of species in the Metarhizium anisopliae complex. Initially the phylogenetic analysis of 5' strains by sequencing of the 59' end of the TEF-1α gene region revealed seven species within M. anisopliae sensu lato and two varieties outside this complex. Because initial studies on MS profiles from different cell types showed that mycelial fragments or conidia produced on nutrient-poor medium may yield too much background noise, all subsequent spectrometric analyses were performed with acidhydrolyzed conidia from 10-12 d old PDA cultures. The initial MALDI-TOF reference library included protein spectral profiles from nine taxonomically distinct, molecularly identified isolates sharing high genetic homology with the ex-type or ex-epitype isolates of these taxa in Metarhizium. A second reference library added one isolate each for M. anisopliae sensu stricto and M. robertsii. The second, larger reference library (including 11 taxa) allowed nearly perfect MALDI-TOF matching of DNA-based species identification for the 40 remaining isolates molecularly recognized as M. anisopliae sensu stricto (n = 19), M. robertsii (n = 6), M. majus (n = 3), M. lepidiotae (n = 1), M. acridum (n = 3), M. flavoviride var. pemphigi (n = 1), plus seven unidentified strains (six of them phylogenetically close to M. anisopliae sensu stricto and one outside the Metarhizium pingshaense-anisopliae-robertsii-brunneum clade). Due to the increasing frequency of phylogenetically (genomically) based taxonomic revisions of fungi, this approach is especially useful for culture collections, because once the protein profiles of Metarhizium isolates are obtained taxonomic updating of MALDI

  10. Pathogenicity of three entomopathogenic fungi, Metarhizium anisopliae, Beauveria bassiana, and Paecilomyces lilacinus, to Tetranychus kanzawai infesting papaya seedlings

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    Yayan Sanjaya

    2016-09-01

    Full Text Available Tetranychus kanzawai is an economically important polyphagous mite species in East and South Asia. In the Philippines, it commonly infests cassava and papaya plants. The mites attack and severely damage the older leaves of papaya trees as well as seedlings. Its serious damage causes the leaves to dry up, thus, reducing the photosynthetic activity of the plant. Three entomopathogenic fungal isolates were tested on mites under greenhouse conditions using treated papaya seedlings following a completely randomized design. The mites tested were examined under a dissecting scope to determine the causal agent and to confirm mortality. The LT50 of Metarhizium anisopliae, Beauveria bassiana and Paecilomyces lilacinus on T. kanzawai were estimated. Our results indicate that among these entomopathogenic fungi, the Metarhizium anisopliae Ma6 isolate (LT50= 3.00 days has potentiality for the control of T. kanzawai.

  11. Susceptibility of different life stages of Ornithodoros lahorensis to entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana.

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    Tavassoli, Mousa; Malekifard, Farnaz; Soleimanzadeh, Ali; Pourseyed, Seyed Hassan; Bernousi, Iraj; Mardani, Karim

    2012-10-01

    The use of chemicals for the control of arthropod pests can be problematic due to the potential for both environmental contamination and resistance development. As a result, there is an increasing interest in nonchemical alternatives, such as biocontrol by entomopathogenic fungi. In the present study, three strains of Metarhizium anisopliae (V245, 3247, and 4456) and one strain of Beauveria bassiana (LM 3.2) were evaluated under laboratory conditions for their virulence towards three life stages of Ornithodoros lahorensis. Groups of eggs, larvae, and adult ticks were treated by immersion in two different suspensions (10(5) and 10(7) conidia/ml) of each fungal strain. All treatment and control groups were observed during a 3-week period, and the hatchability of eggs and mortality percentage of larvae and adult ticks were assessed. The effect of fungal strains on egg hatchability and larva and adult mortality was significant and dose dependent compared to the control groups (P fungi potential biological control agents of O. lahorensis reducing the use of chemical acaricides.

  12. Effect of Biofertilizers and Neem Oil on the Entomopathogenic Fungi Beauveria bassiana (Bals. Vuill. and Metarhizium anisopliae (Metsch. Sorok

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    Hirose Edson

    2001-01-01

    Full Text Available The in vitro fungitoxic effect of three biofertilizers, E.M.-4, Multibion Ô and Supermagro used in organic agriculture and the neem oil (Azadirachta indica A. Juss on the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana was studied. These products were mixed in a medium where the two fungi were inoculated, and germination, vegetative growth and conidiogenesis were assessed. The biofertilizers Supermagro and E.M.- 4 showed to be less toxic for the two fungi whereas MultibionÔ caused major inhibition on M. anisopliae, with reductions in germination (-37.74%, colony diameter (-30.26% and conidiogenesis (-42.62%. Neem oil promoted a larger negative effect on B. bassiana, inhibiting germination (-45.27%, colony diameter (-36.62% and conidiogenesis (-84.93%.

  13. Evaluation of the entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, and Isaria fumosorosea for control of Sitophilus oryzae.

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    Kavallieratos, Nickolas G; Athanassiou, Christos G; Aountala, Maria M; Kontodimas, Demetrius C

    2014-01-01

    The entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, and Isaria fumosorosea were tested against the stored-grain pest Sitophilus oryzae. The fungi were isolated from the soil (from three locations in Attica, Greece: B. bassiana from Tatoion, M. anisopliae from Marathon, and I. fumosorosea from Aghios Stefanos) using larvae of Galleria mellonella as bait. Suspensions of 2.11 × 10(7) and 2.11 × 10(8), 1.77 × 10(7) and 1.77 × 10(8), and 1.81 × 10(7) and 1.81 × 10(8) conidia per ml of B. bassiana, M. anisopliae, and I. fumosorosea, respectively, were applied by three treatments: (i) sprayed on food and set in petri dishes with adults of S. oryzae, (ii) sprayed on adults of S. oryzae and set in petri dishes without food, and (iii) sprayed on adults of S. oryzae and set in petri dishes with food. The observed mortality of S. oryzae adults during the overall exposure period for the lowest, as well as for the highest, concentrations of B. bassiana, M. anisopliae, and I. fumosorosea ranged from 0 to 100%. Concentration was, in most of the cases tested, a critical parameter that determined the "speed of kill" of the exposed insect species for B. bassiana and M. anisopliae. Conversely, concentration was not that critical for I. fumosorosea, and survival was high in some of the combinations tested, even after 14 days of exposure. Both in the highest and the lowest concentrations of fungi, the mortality of S. oryzae adults was higher when the fungi were applied on adults than when they were applied on food. Higher mortality was observed when food was absent than when food was present, in most of the cases tested. The high efficacy levels recorded in the current study indicate that the tested fungi could be effective biocontrol agents against S. oryzae.

  14. Genome-assisted development of nuclear intergenic sequence markers for entomopathogenic fungi of the Metarhizium anisopliae species complex.

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    Kepler, R M; Rehner, S A

    2013-03-01

    Entomopathogenic fungi in the genus Metarhizium are useful for biological control programmes against economically important arthropod pests worldwide. However, understanding the true diversity and ecology of these organisms is hampered by convergent morphologies between species. The application of molecular techniques has enabled greater resolution of species than allowed by morphology alone. In particular, the commonly used biocontrol agent M. anisopliae was found to be a species complex composed of nine species. This prior work was conducted with commonly used markers in fungal phylogenetics (BTUB, RPB1, RPB2 and TEF), which likely under-represent diversity in the M. anisopliae complex. Using sequence data from nuclear genomes of M. acridum and M. robertsii we identified regions of conserved gene synteny and developed primers to amplify intergenic regions of seven loci. Using ex-type and authenticated tissue specimens for species in the M. anisopliae complex, we demonstrate that sequence data derived from intergenic loci is more variable and phylogenetically informative than previously available markers. These new markers will facilitate investigations at or below the species level for the M. anisopliae complex. The method of marker development employed here should be extendable to any group with sufficiently divergent genome data available.

  15. Susceptibility of adults of the cerambycid beetle Hedypathes betulinus to the entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, and Purpureocillium lilacinum.

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    Schapovaloff, M E; Alves, L F A; Fanti, A L; Alzogaray, R A; López Lastra, C C

    2014-01-01

    The cerambycid beetle Hedypathes betulinus (Klug) (Coleoptera: Cerambycidae) causes severe damage to yerba mate plants (Ilex paraguariensis (St. Hilaire) (Aquifoliales: Aquifoliaceae)), which results in large losses of production. In this study, the pathogenicity of entomopathogenic fungi of the species Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Hypocreales: Cordycipitaceae), Metarhizium anisopliae sensu lato (Metschnikoff) Sorokin (Hypocreales: Clavicipitaceae), and Purpureocillium lilacinum (Thom) Luangsa-ard, Hywel-Jones, Houbraken and Samson (Hypocreales: Ophiocordycipitaceae) on yerba mate were evaluated. Fifteen isolates of B. bassiana, two of M. anisopliae, and seven of P. lilacinum on H. betulinus adults were analyzed under laboratory conditions. The raw mortality rate caused by B. bassiana isolates varied from 51.1 to 86.3%, and their LT50 values varied between 8.7 and 13.6 d. The isolates of M. anisopliae caused 69.6‒81.8% mortality, and their LT50 values varied between 7.4 and 7.9 d. In contrast, isolates of P. lilacinum were not pathogenic. M. anisopliae and B. bassiana isolates were pathogenic against H. betulinus adults, suggesting that they may be useful in biological control programs for insect pests of yerba mate.

  16. EFEITO FUNGITÓXICO DO ÓLEO DE NIM SOBRE Metarhizium anisopliae var. acridum e Metarhizium anisopliae var. anisopliae

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    Álison Bruno da Silva Santos

    2009-01-01

    Full Text Available Plague control is based almost exclusively on application of chemical substances, however these products are toxic to men and animals and cause odd effects on environment quality. In Plague Integrated Management (PIM, the use of selected insecticides and entomopathogenic fungi should be considered as one viable strategy for plague control in agriculture. This work aimed to evaluate, in laboratory, the compatibility of the entomopathogenic fungi Metarhizium anisopliae var. acridum and Metarhizium anisopliae var. anisopliae with the oil of Nim. The addition of the product was made to the potato-dextrose-agar medium still liquid (±45°C, in a way that the final concentration obeyed 50% of the producer's recommendation. After fungi inoculation, the dishes were incubated in a cimatized room at 28°C, photophase of 12 hours and relative humidity of 75±5% for 12 day period. The number of conidia per colonie was counted with a Neubauer chamber. Statistic delineament was entirely in random, with two treatments (PDA with insecticide, and a control group (PDA without insecticide, and 9 repetitions for each treatment. The results showed that the insecticide inhibited conidial production in Metarhizium anisopliae var. anisopliae strains when compared to the control group. The diameter of Metarhizium anisopliae var. acridum colonies suffered significative reduction in its size, compared to control. The tested insecticide, in the concentration and formulation used, presented compatibility with the tested strains.

  17. Potential of two entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae (Coleoptera: Scarabaeidae), as biological control agents against the June beetle.

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    Erler, Fedai; Ates, A Ozgur

    2015-01-01

    The aim of this study was to evaluate the effectiveness of the entomopathogenic fungi (EPF), Beauveria bassiana (Bals.) Vuill. (Deuteromycotina: Hyphomycetes) strain PPRI 5339 [BroadBand, an emulsifiable spore concentrate (EC) formulation] and Metarhizium anisopliae (Metsch.) Sorokin (Hypocreales: Clavicipitaceae) strain F52 [Met52, both EC and granular (GR) formulations] against the larvae of Polyphylla fullo (L.) (Coleoptera: Scarabaeidae). Larvicidal bioassays were performed in foam boxes (100 by 75 by 50 cm; length by width by height), containing moist soil medium with some humus and potato tubers as food. Although the B. bassiana product (min. 4 × 10(9) conidia/ml) was applied at 100, 150, and 200 ml/100 l water; M. anisopliae strain F52 was applied at 500, 1,000, and 1,500 g/m(3) of moist soil medium for GR (9 × 10(8) cfu/g) and 75, 100, and 125 ml/100 l water for EC (5.5 × 10(9) conidia/ml) formulation. Both fungi were pathogenic to larvae of the pest; however, young larvae (1st and 2nd instars) were more susceptible to infection than older ones (3rd instar). Mortality rates of young and older larvae varied with conidial concentration of both fungi and elapsed time after application. The B. bassiana product was more effective than both of the formulations of the M. anisopliae product, causing mortalities up to 79.8 and 71.6% in young and older larvae, respectively. The highest mortality rates of young and older larvae caused by the M. anisopliae product were 74.1 and 67.6% for the GR formulation, 70.2 and 61.8% for the EC formulation, respectively. These results may suggest that both fungi have potential to be used for management of P. fullo.

  18. Persistence of Brazilian isolates of the entomopathogenic fungi Metarhizium anisopliae and M. robertsii in strawberry crop soil after soil drench application

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    Castro, Thiago; Mayerhofer, Johanna; Enkerli, Jürg

    2016-01-01

    Establishment, persistence and local dispersal of the entomopathogenic fungi Metarhizium anisopliae (ESALQ1037) and M. robertsii (ESALQ1426) (Ascomycota: Hypocreales) were investigated in the soil and rhizosphere following soil drench application in strawberries between 2012 and 2013 at a single ...

  19. Anopheline and culicine mosquitoes are not repelled by surfaces treated with the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana

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    Mnyone Ladslaus L

    2010-08-01

    Full Text Available Abstract Background Entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana, are promising bio-pesticides for application against adult malaria mosquito vectors. An understanding of the behavioural responses of mosquitoes towards these fungi is necessary to guide development of fungi beyond the 'proof of concept' stage and to design suitable intervention tools. Methods Here we tested whether oil-formulations of the two fungi could be detected and avoided by adult Anopheles gambiae s.s., Anopheles arabiensis and Culex quinquefasciatus. The bioassays used a glass chamber divided into three compartments (each 250 × 250 × 250 mm: release, middle and stimulus compartments. Netting with or without fungus was fitted in front of the stimulus compartment. Mosquitoes were released and the proportion that entered the stimulus compartment was determined and compared between treatments. Treatments were untreated netting (control 1, netting with mineral oil (control 2 and fungal conidia formulated in mineral oil evaluated at three different dosages (2 × 1010, 4 × 1010 and 8 × 1010 conidia m-2. Results Neither fungal strain was repellent as the mean proportion of mosquitoes collected in the stimulus compartment did not differ between experiments with surfaces treated with and without fungus regardless of the fungal isolate and mosquito species tested. Conclusion Our results indicate that mineral-oil formulations of M. anisopliae and B. bassiana were not repellent against the mosquito species tested. Therefore, both fungi are suitable candidates for the further development of tools that aim to control host-seeking or resting mosquitoes using entomopathogenic fungi.

  20. Cuticle Fatty Acid Composition and Differential Susceptibility of Three Species of Cockroaches to the Entomopathogenic Fungi Metarhizium anisopliae (Ascomycota, Hypocreales).

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    Gutierrez, Alejandra C; Gołębiowski, Marek; Pennisi, Mariana; Peterson, Graciela; García, Juan J; Manfrino, Romina G; López Lastra, Claudia C

    2015-04-01

    Differences in free fatty acids (FFAs) chemical composition of insects may be responsible for susceptibility or resistance to fungal infection. Determination of FFAs found in cuticular lipids can effectively contribute to the knowledge concerning insect defense mechanisms. In this study, we have evaluated the susceptibility of three species of cockroaches to the entomopathogenic fungi Metarhizium anisopliae (Metschnikoff) Sorokin by topical application. Mortality due to M. anisopliae was highly significant on adults and nymphs of Blattella germanica L. (Blattodea: Blattellidae). However, mortality was faster in adults than in nymphs. Adults of Blatta orientalis L. (Blattodea: Blattidae) were not susceptible to the fungus, and nymphs of Blaptica dubia Serville (Blattodea: Blaberidae) were more susceptible to the fungus than adults. The composition of cuticular FFAs in the three species of cockroaches was also studied. The analysis indicated that all of the fatty acids were mostly straight-chain, long-chain, saturated or unsaturated. Cuticular lipids of three species of cockroaches contained 19 FFAs, ranging from C14:0 to C24:0. The predominant fatty acids found in the three studied species of cockroaches were oleic, linoleic, palmitic, and stearic acid. Only in adults of Bl. orientalis, myristoleic acid, γ-linolenic acid, arachidic acid, dihomolinoleic acid, and behenic acid were identified. Lignoceric acid was detected only in nymphs of Bl. orientalis. Heneicosylic acid and docosahexaenoic acid were identified in adults of Ba. dubia.

  1. Ethanol production from chitosan by the nematophagous fungus Pochonia chlamydosporia and the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana.

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    Aranda-Martinez, Almudena; Naranjo Ortiz, Miguel Ángel; Abihssira García, Isabel Sofía; Zavala-Gonzalez, Ernesto A; Lopez-Llorca, Luis Vicente

    2017-11-01

    Chitin is the second most abundant biopolymer after cellulose and virtually unexplored as raw material for bioethanol production. In this paper, we investigate chitosan, the deacetylated form of chitin which is the main component of shellfish waste, as substrate for bioethanol production by fungi. Fungal parasites of invertebrates such as the nematophagous Pochonia chlamydosporia (Pc) or the entomopathogens Beauveria bassiana (Bb) and Metarhizium anisopliae (Ma) are biocontrol agents of plant parasitic nematodes (eg. Meloidogyne spp.) or insect pests such as the red palm weevil (Rhynchophorus ferrugineus). These fungi degrade chitin-rich barriers for host penetration. We have therefore tested the chitin/chitosanolytic capabilities of Pc, Bb and Ma for generating reducing sugars using chitosan as only nutrient. Among the microorganisms used in this study, Pc is the best chitosan degrader, even under anaerobic conditions. These fungi have alcohol dehydrogenase (ADH) and pyruvate decarboxylase (PDC) encoding genes in their genomes. We have therefore analyzed their ethanol production under anaerobic conditions using chitosan as raw material. P. chlamydosporia is the largest ethanol producer from chitosan. Our studies are a starting point to develop chitin-chitosan based biofuels. Copyright © 2017 Elsevier GmbH. All rights reserved.

  2. Development of Metarhizium anisopliae and Beauveria bassiana formulations for control of malaria mosquito larvae

    NARCIS (Netherlands)

    Bukhari, S.T.; Takken, W.; Koenraadt, C.J.M.

    2011-01-01

    Background The entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana have demonstrated effectiveness against anopheline larvae in the laboratory. However, utilising these fungi for the control of anopheline larvae under field conditions, relies on development of effective means of app

  3. Pyrethroid resistance in Anopheles gambiae leads to increased susceptibility to the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana

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    Knols Bart GJ

    2010-06-01

    Full Text Available Abstract Background Entomopathogenic fungi are being investigated as a new mosquito control tool because insecticide resistance is preventing successful mosquito control in many countries, and new methods are required that can target insecticide-resistant malaria vectors. Although laboratory studies have previously examined the effects of entomopathogenic fungi against adult mosquitoes, most application methods used cannot be readily deployed in the field. Because the fungi are biological organisms it is important to test potential field application methods that will not adversely affect them. The two objectives of this study were to investigate any differences in fungal susceptibility between an insecticide-resistant and insecticide-susceptible strain of Anopheles gambiae sensu stricto, and to test a potential field application method with respect to the viability and virulence of two fungal species Methods Pieces of white polyester netting were dipped in Metarhizium anisopliae ICIPE-30 or Beauveria bassiana IMI391510 mineral oil suspensions. These were kept at 27 ± 1°C, 80 ± 10% RH and the viability of the fungal conidia was recorded at different time points. Tube bioassays were used to infect insecticide-resistant (VKPER and insecticide-susceptible (SKK strains of An. gambiae s.s., and survival analysis was used to determine effects of mosquito strain, fungus species or time since fungal treatment of the net. Results The resistant VKPER strain was significantly more susceptible to fungal infection than the insecticide-susceptible SKK strain. Furthermore, B. bassiana was significantly more virulent than M. anisopliae for both mosquito strains, although this may be linked to the different viabilities of these fungal species. The viability of both fungal species decreased significantly one day after application onto polyester netting when compared to the viability of conidia remaining in suspension. Conclusions The insecticide

  4. Evaluation of Pathogenicity of the Fungi Metarhizium anisopliae and Beauveria bassiana in Hazelnut Weevil (Curculio nucum L., Coleoptera, Curculionidae) Larvae.

    Science.gov (United States)

    Cheng, Yunqing; Liu, Ting; Zhao, Yixin; Geng, Wanting; Chen, Longtao; Liu, Jianfeng

    2016-12-01

    The nut weevil (Curculio nucum) is one of the most important and widespread pests in hazelnut orchards. In order to screen entomopathogenic fungal strains with high virulence against C. nucum, the growth rate, sporulation, and cumulative mortality of different Metarhizium anisopliae and Beauveria bassiana strains were investigated, and the process by which M. anisopliae CoM 02 infects C. nucum larvae was observed using scanning electron microscopy. The results indicated that the growth rate and sporulation of different fungal strains significantly differed. Thirteen days after inoculation with M. anisopliae CoM 02, the cumulative mortality of C. nucum larvae reached 100 %, which was considerably higher than that of the other five strains. As the most virulent of the six test strains, the cadaver rate, LT50, and LT90 of M. anisopliae CoM 02 were 93.4 %, 7.05 and 11.90 days, respectively. Analysis of the infection process by scanning electron microscopy showed that the spore attachment, hyphal germination, hyphal rapid growth, and sporulation of M. anisopliae CoM 02 occurred on the 3rd, 5th, 7th, and 11th day after inoculation, respectively, indicating that the infection cycle takes approximately 11 days. This finding suggests that the highly virulent M. anisopliae plays an important role in the biocontrol of C. nucum in China.

  5. The effect of leaf biopesticide Mirabilis jalapa and fungi Metarhizium anisopliae to immune response and mortality of Spodoptera exigua instar IV

    Science.gov (United States)

    Suryani, A. Irma; Anggraeni, Tjandra

    2014-03-01

    Spodoptera exigua is one of insect causing damage in agriculture sector. This insect can be controlled by a natural biopesticide by combining two agents of biological control, biopesticides Mirabilis jalapa and entomopathogenic fungi Metarhizium anisopliae, considered to be virulent toward a wide range of insects. The objective of research was to determine the effect of biopesticides M. jalapa and fungi M. anisopliae against immune system and mortality of S. exigua. This research used a complete randomized block design with five concentrations Mirabilis jalapa and optimum dose of M. anisopliae. A high dose of M. jalapa (0.8% w/v) is the most effective one to decrease total haemocytes especially granulocyt and plasmatocyt (cellular immune) and decrease the concentration of lectin (humoral immune) from S. exigua (p < 0.05). The combination of M. jalapa (0, 8% w/v) and lethal dose of M. anisopliae 2.59 × 107 spore/ml were significant to increase mortality of S. exigua within 48 hours (p < 0.05).

  6. Advances and Perspectives of the use of the entomopathogenic fungi beauveria bassiana and metarhizium anisopliae for the control of arthropod pests in poultry production

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    DGP Oliveira

    2014-03-01

    Full Text Available Global poultry production is plagued by a wide variety of arthropods. The problems associated with their chemical control have led to an increasing search for control alternatives, and entomopathogenic fungi seem to be a promising strategy. Despite the large number of insects and mites considered as important pests in animal production, studies on the use of entomopathogenic fungi for their control are still scarce compared with agricultural pests, particularly in Brazil. This article reviews some damages and control aspects of the main arthropod pests that affect Brazilian poultry production, including house flies, lesser mealworms, and feather mites, by the use of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae. Studies published in the last 20 years were reviewed, and the main problems and limitations of that pest-control strategy are discussed.

  7. A Quick Method for Metarhizium anisopliae Isolation from Cultural Soils

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    M. A. Tajick Ghanbary

    2009-01-01

    Full Text Available Problem statement: Fungi are one of the most active members in biological community of cultural soils. Many saprophyte and facultative parasitic fungi live in soil. Metarhizium anisopliae, one of the most famous soil inhabitant entomopathogens has a virulence potential on plant and animal pests. Approach: Introducing a new method for its isolation from soil was an applied method to find it without any limitation. Metarhizium anisopliae shifts to saprophytic phase and remain alive within soil in absence of susceptible host. As a shortcut, we can transfer the fungus from soil to lab by culturing soil suspension. One hundred cultural soil samples from different regions of Iran were tested to finding Metarhizium isolates. Culturing 1:5000-1:10000 soil suspension on artificial medium containing necessary macro and micronutrients for fungal growth were resulted in isolation. Metarhizium anisopliae isolates were harvested seven days after culturing the suspensions. All isolates were inoculated in 50 mL PDB in destruxin production assay and 7 days later broth medium was filtrated by using filter paper. Culture filtrates were extracted and in bioassays they were sprayed on larva of citrus leaf miner. Results: Nine isolates of Metarhizium anisopliae were harvested. Microscopic studies showed that morphological features had complete coincidence with valid descriptions of the fungus. Bioassay confirmed that all harvested isolates secrete active and effective destruxin in broth. Conclusion: Isolation of Metarhizium by culturing the soil suspension, a useful method for more studies of the entomopathogen at different geographical regions. Native populations of this fungus had special importance in local biological control programs. This procedure was a costs- and time-effective method for pathogen isolation.

  8. External development of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae in the subterranean termite Heterotermes tenuis Desenvolvimento dos fungos entomopatogênicos Beauveria bassiana E Metarhizium anisopliae no cupim subterrâneo Heterotermes tenuis

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    Alcides Moino Jr.

    2002-06-01

    Full Text Available The subterranean termite Heterotermes tenuis is one of the main pests of sugarcane and eucalyptus in Brazil, and the use of entomopathogenic fungi, alone or associated to chemicals, is an efficient and environmentally favorable method for its control. Studies related to the fungal development on these insects are important due to the effect of insect behavior on entomopathogens. The objective of this work was to describe the external development of Beauveria bassiana and Metarhizium anisopliae on H. tenuis using Scanning Electron Microscopy (SEM, determining the duration of the different phases of fungal infection. Two fixation techniques for preparing SEM samples were also evaluated. Worker specimens of H. tenuis were inoculated with a 1 x 10(9 conidia mL-1 suspension of the fungi and maintained at 25±1ºC and 70±10% relative humidity. Insects were collected from 0 to 144 hours after inoculation and prepared on SEM stubs for each of the two fixation techniques. The results obtained with the two techniques were compared and duration of the different phases of the infection process were estimated from SEM observations and compared for three fungal isolates. B. bassiana and M. anisopliae have similar development cycles on the termite, but some important differences exist. The penetration, colonization and conidiogenesis phases are relatively faster for M. anisopliae than for B. bassiana, which results in a faster rate of insect mortality. The fixation technique with OsO4 vapor is suitable for preparation of insects to be used in SEM observation of the developmental stages of entomopathogenic fungi.O cupim subterrâneo Heterotermes tenuis , uma das principais pragas da cana-de-açúcar e eucalipto no Brasil, e o uso de fungos entomopatogênicos, isoladamente ou associados a produtos químicos, é um método eficiente e ambientalmente seguro para seu controle. Estudos relacionados ao desenvolvimento fúngico nestes insetos são importantes devido

  9. PROTEOMIC ANALYSIS OF ALLERGENS FROM METARHIZIUM ANISOPLIAE

    Science.gov (United States)

    IntroductionThe goal of this project is the identification and characterization of allergens from the fungus Metarhizium anisopliae, using mass spectrometry (MS). The US EPA, under the "Children at Risk" program, is currently addressing the problem of indoor fungal bioaer...

  10. INFECTIVITY OF METARHIZIUM ANISOPLIAE IN GRASS SHRIMP EMBRYOS

    Science.gov (United States)

    Developing embryos of the estuarine grass shrimp, Palaemonetes pugio, were exposed to Metarhizium anisopliae conidiospores. Attachment of conidiospores was often followed by germination and outgrowth on embryo surface. Penetration of the embryonic envelopes by M. anisopliae allow...

  11. INFECTIVITY OF METARHIZIUM ANISOPLIAE IN GRASS SHRIMP EMBRYOS

    Science.gov (United States)

    Developing embryos of the estuarine grass shrimp, Palaemonetes pugio, were exposed to Metarhizium anisopliae conidiospores. Attachment of conidiospores was often followed by germination and outgrowth on embryo surface. Penetration of the embryonic envelopes by M. anisopliae allow...

  12. Cecropins from Plutella xylostella and Their Interaction with Metarhizium anisopliae.

    Science.gov (United States)

    Ouyang, Lina; Xu, Xiaoxia; Freed, Shoaib; Gao, Yanfu; Yu, Jing; Wang, Shuang; Ju, Wenyan; Zhang, Yuqing; Jin, Fengliang

    2015-01-01

    Cecropins are the most potent induced peptides to resist invading microorganisms. In the present study, two full length cDNA encoding cecropin2 (Px-cec2) and cecropin3 (Px-cec3) were obtained from P. xylostella by integrated analysis of genome and transcriptome data. qRT-PCR analysis revealed the high levels of transcripts of Px-cecs (Px-cec1, Px-cec2 and Px-cec3) in epidermis, fat body and hemocytes after 24, 30 and 36 h induction of Metarhizium anisopliae, respectively. Silencing of Spätzle and Dorsal separately caused the low expression of cecropins in the fat body, epidermis and hemocytes, and made the P.xylostella larvae more susceptible to M. anisopliae. Antimicrobial assays demonstrated that the purified recombinant cecropins, i.e., Px-cec1, Px-cec2 and Px-cec3, exerted a broad spectrum of antimicrobial activity against fungi, as well as Gram-positive and Gram-negative bacteria. Especially, Px-cecs showed higher activity against M. anisopliae than another selected fungi isolates. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed that cecropins exerted the vital morphological alterations to the spores of M. anisopliae. Based on our results, cecropins played an imperative role in resisting infection of M. anisopliae, which will provide the foundation of biological control of insect pests by using cecorpins as a target in the future.

  13. Cecropins from Plutella xylostella and Their Interaction with Metarhizium anisopliae.

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    Lina Ouyang

    Full Text Available Cecropins are the most potent induced peptides to resist invading microorganisms. In the present study, two full length cDNA encoding cecropin2 (Px-cec2 and cecropin3 (Px-cec3 were obtained from P. xylostella by integrated analysis of genome and transcriptome data. qRT-PCR analysis revealed the high levels of transcripts of Px-cecs (Px-cec1, Px-cec2 and Px-cec3 in epidermis, fat body and hemocytes after 24, 30 and 36 h induction of Metarhizium anisopliae, respectively. Silencing of Spätzle and Dorsal separately caused the low expression of cecropins in the fat body, epidermis and hemocytes, and made the P.xylostella larvae more susceptible to M. anisopliae. Antimicrobial assays demonstrated that the purified recombinant cecropins, i.e., Px-cec1, Px-cec2 and Px-cec3, exerted a broad spectrum of antimicrobial activity against fungi, as well as Gram-positive and Gram-negative bacteria. Especially, Px-cecs showed higher activity against M. anisopliae than another selected fungi isolates. Scanning electron microscopy (SEM and transmission electron microscopy (TEM revealed that cecropins exerted the vital morphological alterations to the spores of M. anisopliae. Based on our results, cecropins played an imperative role in resisting infection of M. anisopliae, which will provide the foundation of biological control of insect pests by using cecorpins as a target in the future.

  14. PCR-RFLP analysis of chitinase genes enables efficient genotyping of Metarhizium anisopliae var. anisopliae.

    Science.gov (United States)

    Enkerli, Jürg; Ghormade, Vandana; Oulevey, Catherine; Widmer, Franco

    2009-10-01

    A new genotyping tool has been developed and evaluated for Metarhizium anisopliae var. anisopliae. The tool is based on Restriction Fragment Length Polymorphism (RFLP) analysis of three chitinase genes that are functionally linked to insect-pathogenicity of this fungus. It allowed for discrimination of 14 genotypes among 22 M. anisopliae var. anisopliae strains of a world wide collection. Analyses revealed that the approach may also be applicable to other Metarhizium varieties. The new tool will be useful for genetic characterization of M. anisopliae var. anisopliae strains, and it is applicable for laboratories with limited access to molecular diagnostic equipment.

  15. Beauveria bassiana, Metarhizium anisopliae, and Metarhizium anisopliae var. acridum conidia: tolerance to imbibitional damage

    Science.gov (United States)

    When dry fungal cells are immersed in water, rapid imbibition (water uptake) may compromise the plasma membrane, killing the cell. This study investigated the impact of imbibitional damage (measured in terms of reduced viability) on Beauveria bassiana (Bb), Metarhizium anisopliae (Ma) and M. anisop...

  16. Ação in vitro dos fungos Beauveria bassiana (Bals Vuill e Metarhizium anisopliae (Metsch Sorok sobre ninfas e adultos de Amblyomma cajennense (Fabricius, 1787 (Acari: Ixodidae In vitro action of the fungi Beauveria bassiana (Bals Vuill and Metarhizium anisopliae (Metsch Sorok on ninphs and adults of Amblyomma cajennense (Fabricius, 1787 (Acari: Ixodidae

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    R.C.S. Reis

    2001-10-01

    Full Text Available O trabalho teve por objetivo verificar a mortalidade in vitro de ninfas e adultos de Amblyomma cajennense frente à ação de isolados dos fungos Beauveria bassiana e Metarhizium anisopliae. Foram avaliados três isolados de M. anisopliae (959, 319 e E9 e dois de B. bassiana (986 e 747. As suspensões de conídios foram preparadas a partir de fungos produzidos em meio de arroz, e cada bioensaio foi constituído de quatro tratamentos nas concentrações 10(5, 10(6, 10(7, 10(8 conídios/ml e um grupo-controle. A análise constou da observação do percentual de mortalidade, 15 dias após o tratamento ou após a ecdise de adultos. Foram observadas diferenças significativas entre os tratamentos quanto à mortalidade para todos os isolados e todos os instares. Nos grupos tratados houve aumento considerável na mortalidade à medida que se aumentava a concentração de conídios na suspensão. Conclui-se que todos os isolados testados causaram mortalidade em testes in vitro sobre esses estádios evolutivos, sugerindo o controle do A. cajennense pela ação desses fungos.This work aimed at the evaluation of the in vitro susceptibility of Amblyomma cajennense nimphs and adults to isolates of Beauveria bassiana and Metarhizium anisopliae fungi. Three isolates of M. anisopliae (959, 319 e E9 and two of B. bassiana (986 e 747 were used. Conidia suspensions were made from fungi grown up in a rice culture medium and each test consisted of four treated groups (10(5, 10(6, 10(7, 10(8 conidia/ ml plus a control group. Viability of individuals was assessed 15 days after treatment or after adult ecdisis. Significative differences were found for all treatments and for all stages studied. A large reduction in the viability of ticks was observed, and this effect increased as conidia concentration raised. Based upon the results obtained, it can be concluded that all isolates tested presented an in vitro lethality for the biological stages of A. cajennense evaluated

  17. The Impact of Culture Age, Aeration, and Agitation on the Production of Microsclerotia of the Entomopathogenic Fungus Metarhizium anisopliae Using 100-Liter Fermentors

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    Microsclerotia are desiccation-tolerant, compact hyphal aggregates produced by numerous fungi as overwintering structures. We recently discovered that the entomopathogenic fungus Metarhizium anisopliae produced microsclerotia during liquid culture fermentation. When air-dried microsclerotial granu...

  18. Pyrethroid resistance in Anopheles gambiae leads to increased susceptibility to the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana

    NARCIS (Netherlands)

    Howard, A.F.V.; Koenraadt, C.J.M.; Farenhorst, M.; Knols, B.G.J.; Takken, W.

    2010-01-01

    BACKGROUND: Entomopathogenic fungi are being investigated as a new mosquito control tool because insecticide resistance is preventing successful mosquito control in many countries, and new methods are required that can target insecticide-resistant malaria vectors. Although laboratory studies have

  19. Pyrethroid resistance in Anopheles gambiae leads to increased susceptibility to the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana

    NARCIS (Netherlands)

    Howard, A.F.V.; Koenraadt, C.J.M.; Farenhorst, M.; Knols, B.G.J.; Takken, W.

    2010-01-01

    Background: Entomopathogenic fungi are being investigated as a new mosquito control tool because insecticide resistance is preventing successful mosquito control in many countries, and new methods are required that can target insecticide-resistant malaria vectors. Although laboratory studies have

  20. Biological insect control using Metarhizium anisopliae: morphological, molecular, and ecological aspects

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    Patricia Vieira Tiago

    2014-04-01

    Full Text Available Microbial control of insects is based on the rational use of pathogens to maintain environmentally balanced pest population levels, and Metarhizium anisopliae has been the most studied and most utilized fungal species for that purpose. The natural genetic variability of entomopathogenic fungi is considered one of the principal advantages of microbial insect control. The inter- and intraspecific variability and the genetic diversity and population structures of Metarhizium and other entomopathogenic fungi have been examined using ITS-RFLP, ISSR, and ISSP molecular markers. The persistence of M. anisopliae in the soil and its possible effects on the structures of resident microbial communities must be considered when selecting isolates for biological insect control.

  1. Pyrethroid resistance in Anopheles gambiae leads to increased susceptibility to the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana

    NARCIS (Netherlands)

    Howard, A.F.V.; Koenraadt, C.J.M.; Farenhorst, M.; Knols, B.G.J.; Takken, W.

    2010-01-01

    BACKGROUND: Entomopathogenic fungi are being investigated as a new mosquito control tool because insecticide resistance is preventing successful mosquito control in many countries, and new methods are required that can target insecticide-resistant malaria vectors. Although laboratory studies have pr

  2. Pyrethroid resistance in Anopheles gambiae leads to increased susceptibility to the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana

    NARCIS (Netherlands)

    Howard, A.F.V.; Koenraadt, C.J.M.; Farenhorst, M.; Knols, B.G.J.; Takken, W.

    2010-01-01

    Background: Entomopathogenic fungi are being investigated as a new mosquito control tool because insecticide resistance is preventing successful mosquito control in many countries, and new methods are required that can target insecticide-resistant malaria vectors. Although laboratory studies have pr

  3. Use of Beauveria bassiana and Metarhizium anisopliae for fruit fly control: a novel approach

    Energy Technology Data Exchange (ETDEWEB)

    Toledo, Jorge; Liedo, Pablo, E-mail: jtoledo@ecosur.m [El Colegio de la Frontera Sur, Chiapas (Mexico). Dept. de Entomologia Tropical; Flores, Salvador; Montoya, Pablo [Secretaria de Agricultura, Ganaderia, Desarrollo Rural, Pesca y Alimentacion (SAGARPA), Chiapas (Mexico). Subdireccion de Desarrollo de Metodos; Campos, Sergio E.; Villasenor, Antonio [Secretaria de Agricultura, Ganaderia, Desarrollo Rural, Pesca y Alimentacion (SAGARPA), Chiapas (Mexico). Programa Moscamed. Direccion de Operaciones de Campo

    2006-07-01

    The potential of two species of entomopathogenic fungi, Beauveria bassiana (Bals.) and Metarhizium anisopliae (Met.) Sorokin, as practical fruit fly biocontrol agents is studied. These natural inhabitants of soil are found infecting a wide range of insect species that spend at least one stage of their life cycle in the soil. Sterile flies are used as vectors of the infection. A summary of results from different laboratory and field cage experiments is presented. (MAC)

  4. Production of microsclerotia of the fungal entomopathogen Metarhizium anisopliae and their potential for use as a biocontrol agent for soil-inhabiting insects

    Science.gov (United States)

    Microsclerotia (MS), overwintering structures produced by many plant pathogenic fungi, have not been described for Metarhizium anisopliae. Three strains of M. anisopliae – F52, TM109, and MA1200 – formed MS in shake flask cultures using media with varying carbon concentrations and carbon-to-nitroge...

  5. Efficacy of the entomopathogenic fungi Metarhizium anisopliae in the control of infestation by stable flies Stomoxys calcitrans (L.), under natural infestation conditions.

    Science.gov (United States)

    Cruz-Vazquez, C; Carvajal Márquez, J; Lezama-Gutiérrez, R; Vitela-Mendoza, I; Ramos-Parra, M

    2015-09-15

    The objective of this study was to evaluate the efficacy of an isolate of Metarhizium anisopliae applied by aspersion to control of Stomoxys calcitrans flies in dairy cattle naturally infested. Was applied by aspersion an aqueous formulation of M. anisopliae sensu lato (Ma134), at a concentration of 1×10(8)conidia/ml, four times with seven day intervals, on a group of eight Holstein cows; a control group of eight Holstein cows, received a water solution with Tween 80 (0.1%). The average number of flies per animal was estimated one day before each application, and then daily counts were done in both groups. The effectiveness of the formulation was calculated using the Abbott's formula. At the same time, defensive behaviors of stamp/kicks and tail movements were evaluated daily, estimating relative frequency per hour. The Ma134 formulation had an infestation control efficacy of 73%, taking into consideration the four study weeks. The population reduction effect was observed since the first week post-application (p<0.05), and the effect increased with the subsequent applications. Defensive behaviors were reduced beginning from the first application, reaching a reduction of 66% and 70%, respectively, during the four weeks of study. These results demonstrated the effectiveness of the formulation to control infestation by S. calcitrans, as well as reduce defensive behaviors which involves the infestation.

  6. IDENTIFICATION OF ALLERGENS FROM METARHIZIUM ANISOPLIAE USING MASS SPECTROMETRY

    Science.gov (United States)

    Background The U.S. EPA, under the "Children at Risk" Program, is currently addressing the problem of indoor fungal bioaerosol contamination. The fungus Metarhizium Anisopliae has been used as a bio-pesticide for insect control since the 1800's. Recent studies have shown t...

  7. Genetic diversity of Metarhizium anisopliae var. anisopliae in southwestern British Columbia.

    Science.gov (United States)

    Inglis, G Douglas; Duke, Grant M; Goettel, Mark S; Kabaluk, J Todd

    2008-05-01

    The abundance and genetic diversity of the entomopathogenic fungus, Metarhizium anisopliae var. anisopliae, in southwestern British Columbia (BC) and southern Alberta was examined. The fungus was found to be widespread in soil throughout southwestern BC, and was recovered from 56% of 85 sample sites. In contrast to southwestern BC, no M. anisopliae isolates were recovered in southern Alberta. An automated fluorescent amplified fragment length polymorphism (AFLP) method was used to examine genetic diversity. In excess of 200 isolates were characterized. The method identified 211 polymorphic amplicons, ranging in size from approximately 92 to 400 base pairs, and it was found to be reproducible with a resolution limit of 86.2% similarity. The AFLP method distinguished Metarhizium from other entomopathogenic fungal genera, and demonstrated considerable genetic diversity (25 genotypes) among the reference strains of M. anisopliae isolates examined (i.e. recovered from various substrates and geographical locations). Although 13 genotypes of M. anisopliae var. anisopliae were recovered from southwestern BC soils, the vast majority of isolates (91%) belonged to one of two closely-related genotypes. Furthermore, these two genotypes predominated in urban, agricultural and forest soils. The reasons for the limited diversity of M. anisopliae var. anisopliae in southwestern BC are uncertain. However, findings of this study are consistent with island biogeography theory, and have significant implications for the development of this fungus for microbial control of pest insects.

  8. Selectivity of Metarhizium anisopliae and Beauveria bassiana (Hypocreales: Clavicipitaceae) on adults of Cotesia flavipes (Hymenoptera: Braconidae).

    Science.gov (United States)

    Rossoni, Camila; Loureiro, Elisângela De Souza; Pereira, Fabricio Fagundes; Kassab, Samir Oliveira; Costa, Daniele Perassa; Barbosa, Rogério Hidalgo

    2014-01-01

    Understanding mortality patterns and interactions between entomopathogenic fungi and parasitoids is important to improve insect biological control programs. The aim of this study was to evaluate the effect of Metarhizium anisopliae (Metschnikoff, 1879) Sorokin, 1833 and Beauveria bassiana (Balsamo) Vuillemin, 1912 (Hypocreales: Clavicipitaceae) on adults of Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae) with biological insecticides Biometha WP Plus (M. anisopliae), Biovéria G (B. bassiana), Boverril WP (B. bassiana), Metarril WP (M. anisopliae), and Metie WP (M. anisopliae) at concentrations of 1 x 10(9) conidia (con).mL(-1), 5 x 10(9) con.ml(-1), and 10 x 10(9) con.ml(-1). In the experimental, 10 females of C. flavipes were packed in disposable cups capped with a contact surface (filter paper, 9 cm2) treated with commercial product. The experimental design was completely randomized, with 16 treatments and five replicates of 10 females each. Mortality was assessed at 24, 48, 72, 96, and 120 hours after exposition (HAE) of the products. In general, B. bassiana and M. anisopliae in the concentrations of 1 x 10(9) con.ml(-1), 5 x 10(9) con.ml(-1), and 10 x 10(9) con.ml(-1) can't affect C. flavipes females because the peak of mortality in treatments with bioinsecticides was similar to the control and this demonstrated the selectivity of fungi B. bassiana and M. anisopliae on C. flavipes females.

  9. Desenvolvimento dos fungos Metarhizium anisopliae (Metschnikoff, 1879 Sorokin, 1883 E Beauveria bassiana (Balsamo Vuillemin, 1912 sobre Ctenocephalides felis felis (Bouché, 1835 Development of the fungi Metarhizium anisopliae (Metschnikoff, 1879 Sorokin, 1883 and Beauveria bassiana (Balsamo Vuillemin, 1912 on the Ctenophephalides felis felis (Bouché, 1835

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    Denise R. De Melo

    2007-09-01

    Full Text Available A pulga Ctenocephalides felis felis é um parasita causador dermatites alérgicas e também pode transmitir diversos agentes etiológicos aos animais domésticos e aos homens. O objetivo deste trabalho foi verificar o desenvolvimento do fungo sobre a cutícula da pulga, através da microscopia eletrônica de varredura. Os isolados fúngicos testados foram o Metarhizium anisopliae 959 e Beauveria bassiana 986, ambos na concentração 10(8 conídios/ml. Após a exposição dos isolados fúngicos no período de duas, 15, 26 e 96 horas , o material foi processado para a microscopia eletrônica de varredura. Com a obtenção das micrografias, pode-se observar que com 2 horas após exposição aos fungos, os conídios estavam aderidos por toda a cutícula, situando-se preferencialmente nas membranas intersegmentais do abdome. Com 15 horas observou-se a formação do tubo de germinação e a cabeça do apressório e após 26 horas foi possível observar as ramificações e o engrossamento das hifas sobre a cutícula das pulgas. Os resultados indicam que os fungos testados foram capazes de se desenvolver sobre a cutícula de C. f. felis.The flea Ctenocephalides felis felis is a parasite that causes allergic dermatitis and also may transmit etiologic agents to domestic animals and humans. This study investigated by scanning electron microscopy the development of entomopathogenic fungi on flea cuticle. Fleas were exposed to conidia (10(8 ml-1 of Metarhizium anisopliae (isolate 959 or Beauveria bassiana (isolate 986. Following standard protocols for electron microscopy, the specimens were prepared 2, 15, 26 and 96 h after infection. The micrography revealed that 2 h after fungus exposure, conidia attachments encompassed the entire flea cuticle, especially on abdominal intersegmental membranes. The emergence of germ tubes and appressoria formation occurred at 15 h, thickening and branching of hyphae on the flea cuticle was noted at 26 h. Therefore, both of

  10. Efeito de Metarhizium anisopliae (Metsch. Sorok. e Beauveria bassiana (Bals. Vuill. sobre adultos de Oomyzus sokolowskii (Kurdjumov (Hymenoptera: Eulophidae = Effect of Metarhizium anisopliae (Metsch. Sorok. and Beauveria bassiana (Bals. Vuill. on adults of Oomyzus sokolowskii (Kurdjumov (Hymenoptera: Eulophidae

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    Hugo José Gonçalves dos Santos Júnior

    2006-04-01

    Full Text Available Avaliou-se a seletividade dos fungos Metarhizium anisopliae (Metsch. Sorok. e Beauveria bassiana (Bals. Vuill. sobre adultos do parasitóide Oomyzus sokolowskii (Kurdjumov. Foram utilizados os isolados Esalq 447 de B. bassiana e E9 de M. anisopliae, na concentração de 107 conídios mL-1. Os resultados mostraram que B. bassiana e M. anisopliae não reduziram a longevidade média do parasitóide. B. bassiana proporcionou porcentagem de mortalidade total de 26% e porcentagem de mortalidade confirmada de 21%, já M.anisopliae causou mortalidade total de 15% e confirmada de 9%, demonstrando que M. anisopliae foi menos agressivo. Portanto, em função dos resultados apresentados, a implementação no manejo integrado de P. xylostella com M. anisopliae, B. bassiana e O.sokolowskii pode ser uma excelente alternativa para otimizar o sistema produtivo das brássicas.The selectivity of the fungi Metarhizium anisopliae (Metsch. Sorok. and Beauveria bassiana (Bals. Vuill. to adults of the parasitoid Oomyzus sokolowskii (Kurdjumov was evaluated. The isolates E9 of M. anisopliae and Esalq 447 of B. bassiana were used at the concentration of 107 conidia mL-1. The results showed that B. bassiana and M. anisopliae reduced significantly the mean longevity of the adults about 6.7 and 4.7 days respectively. B. bassiana induced 26% and 21% of total and confirmed mortalities, respectively, while M. anisopliae caused 15% and 9% of total and confirmedmortalities, showing that the isolate of M. anisopliae was less aggressive. Therefore, in function of the presented results the combination of M. anisopliae, B. bassiana, and O. sokolowskii in the integrated management of P. xylostella may be an excellent alternative for optimizing the cabbage growing system.

  11. Severe fungal sclerokeratitis caused by Metarhizium anisopliae: a case report and literature review.

    Science.gov (United States)

    Eguchi, Hiroshi; Toibana, Toshi; Hotta, Fumika; Miyamoto, Tatsuro; Mitamura, Yoshinori; Yaguchi, Takashi

    2015-02-01

    To date, there has been only one published report on the infectious sclerokeratitis caused by Metarhizium anisopliae, which is an entomopathogenic fungus. Regarding corneal infection, three reports have been published to date. Although the prognoses of the corneal infections are favourable, prognosis when scleral infection is involved is very poor. A 76-year-old patient presented with foreign body sensation in the left eye. Microscopic examination with Fungi Flora Y staining of the corneal scraping revealed fungal infection. The conjunctiva was melted by the infection over a wide area. Although intensive medications were administered, an emergency surgery was necessary because scleral thinning, corneal perforation and lens prolapse occurred. The fungal isolate was identified as M. anisopliae by sequencing the internal transcribed spacer region. Herein, we report the second known case worldwide of M. anisopliae sclerokeratitis, and we review the literature related to the ocular infections.

  12. Pure culture of Metarhizium anisopliae LHL07 reprograms soybean to higher growth and mitigates salt stress.

    Science.gov (United States)

    Khan, Abdul Latif; Hamayun, Muhammad; Khan, Sumera Afzal; Kang, Sang-Mo; Shinwari, Zabta Khan; Kamran, Muhammad; Ur Rehman, Shafiq; Kim, Jong-Guk; Lee, In-Jung

    2012-04-01

    Little is known about the role of endophytic fungi against abiotic stresses and isoflavonoids (IF) contents of soybean. In current study, we investigated the role of fungal endophytes on the growth of soybean under salt stress conditions. Pure cultures of nine endophytic fungi were isolated from the roots of field-grown soybean plants, and their culture filtrates were screened on Waito-C and Dongjin-byeo rice cultivars; for identification of plant growth promoting fungal strains. It was observed that fungal isolate GMC-2B significantly promoted the growth of both Waito-C and Dongjin-byeo. GMC-2B was later identified as a new strain of Metarhizium anisopliae LHL07 on the basis of 18S rDNA sequences and phylogenetic analysis. Metarhizium anisopliae LHL07 inoculated soybean plants recorded significantly higher shoot length, shoot fresh and dry biomass, chlorophyll contents, transpiration rate, photosynthetic rate and leaf area; under sodium chloride induced salt stress as compared to non-inoculated control plants. An elevated proline and reduced superoxide dismutase and malondialdehyde contents in M. anisopliae LHL07 inoculated soybean plants demonstrated mitigation of salt induced oxidative stress. Furthermore, reduced abscisic acid and elevated jasmonic acid contents in soybean plants confirmed that lesser stress was convened to M. anisopliae inoculated-plants under salinity stress. We also assessed the role of M. anisopliae interaction on IF biosynthesis of soybean, and found significantly higher IF contents in M. anisopliae inoculated soybean plants. In conclusion, endophytic fungal interactions with soybean can be beneficial to improve soybean quality and quantity under salt affected agricultural systems.

  13. First unusual case of keratitis in Europe due to the rare fungus Metarhizium anisopliae.

    Science.gov (United States)

    Dorin, Josephine; Debourgogne, Anne; Zaïdi, Mohamed; Bazard, Marie-Christine; Machouart, Marie

    2015-05-01

    Metarhizium anisopliae is a fungus utilized worldwide for insect-pest biocontrol. Few M. anisopliae infections have been reported previously. Here, M. anisopliae was isolated from a corneal ulcer in a healthy man. It is the first ocular case in France and Europe of this extremely rare fungus in humans.

  14. Development of Metarhizium anisopliae and Beauveria bassiana formulations for control of malaria mosquito larvae

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    Takken Willem

    2011-02-01

    Full Text Available Abstract Background The entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana have demonstrated effectiveness against anopheline larvae in the laboratory. However, utilising these fungi for the control of anopheline larvae under field conditions, relies on development of effective means of application as well as reducing their sensitivity to UV radiation, high temperatures and the inevitable contact with water. This study was conducted to develop formulations that facilitate the application of Metarhizium anisopliae and Beauveria bassiana spores for the control of anopheline larvae, and also improve their persistence under field conditions. Methods Laboratory bioassays were conducted to test the ability of aqueous (0.1% Tween 80, dry (organic and inorganic and oil (mineral and synthetic formulations to facilitate the spread of fungal spores over the water surface and improve the efficacy of formulated spores against anopheline larvae as well as improve spore survival after application. Field bioassays were then carried out to test the efficacy of the most promising formulation under field conditions in western Kenya. Results When formulated in a synthetic oil (ShellSol T, fungal spores of both Metarhizium anisopliae and Beauveria bassiana were easy to mix and apply to the water surface. This formulation was more effective against anopheline larvae than 0.1% Tween 80, dry powders or mineral oil formulations. ShellSol T also improved the persistence of fungal spores after application to the water. Under field conditions in Kenya, the percentage pupation of An. gambiae was significantly reduced by 39 - 50% by the ShellSol T-formulated Metarhizium anisopliae and Beauveria bassiana spores as compared to the effects of the application of unformulated spores. Conclusions ShellSol T is an effective carrier for fungal spores when targeting anopheline larvae under both laboratory and field conditions. Entomopathogenic fungi formulated with

  15. Optimization of Production Medium for Mass Culture of Metarhizium anisopliae ARS 2231

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    Kang, Seung Woo [Korea University, Seoul (Korea); Moon, Ki Hyuk [Kyungwon University Technology Complex, Songnam (Korea); Yoon, Jeong Weon [Suwon University, Suwon (Korea)

    1999-11-01

    Chemical pesticides which have been commonly used for crop protection have many problems such as high production cost and environmental hazards. Entomogeneous fungi which attack living insects are very powerful means for microbiological insecticide. The purpose of this study is to investigate the culture conditions for mass production of Metarhizium anisopliae ARS 2231 which is a potential microbiological pesticide. The temperature and pH range for optimal cultivation were 28 deg. C and pH 5.0-7.0, respectively. For M. anisopliae ARS 2231, 3.0% (w/v) rice bran, 0.8% (w/v) hydrolyzed casein, and 0.3% (w/v) K{sub 2}HPO{sub 4} were found as the proper nutrients, considering cell mass, enzyme activities, and spore concentration. 16 refs., 10 tabs.

  16. Metarhizium anisopliae as a biological control agent against Hyalomma anatolicum (Acari: Ixodidae).

    Science.gov (United States)

    Suleiman, Elham A; Shigidi, M T; Hassan, S M

    2013-12-15

    In the Sudan, ticks and Tick-borne Diseases (TBDs) with subsequent costs of control and treatment are causing substantial economic loss. Control of ticks is mainly by chemical insecticides. The rising environmental hazards and problem of resistance has motivated research on biological agents as alternative methods of control. The present study aims at controlling livestock ticks using fungi for their unique mode of action besides their ability to adhere to the cuticle, to germinate and penetrate enzymatically. The study was conducted to evaluate the fungus Metarhizium anisopliae for tick control as an alternative mean to chemical acaricides. Pathogenicity of the fungus was tested on different developmental stages of the tick Hyalomma anatolicum. The fungus induced high mortality to flat immature stages. It, also, affected reproductive potential of the females. Egg laid, hatching percent, fertility and moulting percent of immature stages were significantly (p anisopliae to control ticks is discussed.

  17. Laboratory evaluation of Beauveria bassiana and Metarhizium anisopliae in the control of Haemaphysalis qinghaiensis in China.

    Science.gov (United States)

    Ren, Qiaoyun; Chen, Ze; Luo, Jin; Liu, Guangyuan; Guan, Guiquan; Liu, Zhijie; Liu, Aihong; Li, Youquan; Niu, Qingli; Liu, Junlong; Yang, Jifei; Han, Xueqing; Yin, Hong; Luo, Jianxun

    2016-06-01

    Haemaphysalis qinghaiensis, a prevalent tick species in China, is an ectoparasite that preferentially infests small ruminants and can transmit Theileria sp. and Babesia sp. In this study, we evaluated the pathogenicity of individual and mixed infections of the fungi Beauveria bassiana and Metarhizium anisopliae to H. qinghaiensis nymphs. The estimated LC50 for ticks immersed in solutions of B. bassiana, M. anisopliae and a mixture thereof were: 5.88056 × 10(4), 2.65 × 10(4), and 2.85 × 10(4) conidia mL(-1) respectively, and the nymphal mortality ranged from 52 to 100 %. Thus, these results suggest a potential approach for the biocontrol of H. qinghaiensis.

  18. Exposure of Bed Bugs to Metarhizium anisopliae at Different Humidities.

    Science.gov (United States)

    Ulrich, Kevin R; Feldlaufer, Mark F; Kramer, Matthew; St Leger, Raymond J

    2014-12-01

    Bed bugs Cimex lectularius L. were exposed to conidia (spores) of the entomopathogenic fungus Metarhizium anisopliae by feeding, aerosol spray, or contact with a treated surface. Feeding experiments demonstrated that bed bugs were innately susceptible to this fungus. However, only at 98% humidity were mortality rates high, regardless of whether bed bugs were sprayed with a fungal solution or contacted a treated surface. Mortality in treated bed bugs at ambient humidity did not increase when these bed bugs were kept in aggregation with other bed bugs that had recently blood fed to repletion. Based on these laboratory studies, we conclude that M. anisopliae is a poor pathogen for use in control of bed bugs, particularly at humidities that would likely be encountered under field conditions. © 2014 Entomological Society of America.

  19. METARHIZIUM ANISOPLIAE DAN ANDROGRAPHIS PANICULATA TERHADAP SERANGGA BUKAN HAMA SASARAN

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    Dini Yuliani

    2016-04-01

    Full Text Available The objective of this study was to determine the effect of Metarhizium anisopliae and Andrographis paniculata to the natural enemies of Nephotettix virescens and non-target insect pests. This research was conducted in tungro endemic areas in Subang District, West Java in the wet season 2013/2014. The method of research used a split plot design with four replications. The main plot was IR66, Inpari 9, and Ciherang varieties. Subplot was the application of M. anisopliae, A. paniculata, and control. Observations were carried out five times started at nursery (2 weeks after seedling, 14, 28, 42, and 56 days after planting using insect nets, double swing 10 times on each plot observations. The results show M. anisopliae and A. paniculata not adversely affect on non-target insect pests such as Chironomid and natural enemies of N. virescens namely Lycosa pseudoannulata, Cyrtorhinus lividipennis, Sepedon sp., damselfly, Tipulidae sp., Telenomus sp., dragonfly, and Tetrastichus sp.. Efficacy of M. anisopliae and A. paniculata as one of the control strategies that are environmentally friendly and proved not harmful to non-target insect pests.

  20. Pathogenicity of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae against larvae of Batocera davidis%白僵菌和绿僵菌不同菌株对橙斑白条天牛幼虫的致病力

    Institute of Scientific and Technical Information of China (English)

    童文钢

    2012-01-01

    橙斑白条天牛是福建省杨树的一种新害虫,应用不同球孢白僵菌(Beauveria bassiana)和金龟子绿僵菌(Metarhizium anisopliae)菌株对橙斑白条天牛幼虫进行致病力测定,旨在筛选出感染该虫的高致病力菌株,为生物防治提供新的资源.结果表明,接种后,该天牛幼虫的累积死亡率随时间的延长而逐渐增高,接种绿僵菌Ma1291-2和Ma1775菌株14 d后,幼虫的校正死亡率分别为91.69%和86.50%;接种白僵菌BbDp-06菌株14 d后,幼虫校正死亡率达71.05%.绿僵菌Mal291-2和Ma1775菌株对该幼虫的LT50分别为5.73和5.39 d,白僵菌BbDp-06菌株的LT50为8.02 d.Ma1291-2菌株对该幼虫的LC50为3.82×104孢子·mLL-1,白僵菌BbDp-06菌株的LC50为3.88 × 104孢子·mL-1.说明绿僵菌对该幼虫的致病力较强,尤其是Ma1291-2和Ma1775菌株,在橙斑白条天牛的生物防治中将有较好的应用价值.%Batocera davidis Deyrolle is a new pest of poplar in Fujian. In order to screen entomopathogenic fungi strains with high virulence against B. davidis larvae, and bioassay evaluation of ft bassiana and M. anisopliae against B. davidis larvae was carried out through dipping in spore suspension. The results showed that the cumulative corrected mortality of B. davidis larvae was gradually increased with time after inoculated with B. bassiana and M. anisopliae. The corrected mortality of B. davidis larvae on the 14th day after inoculated with Mal291-2 and Mal77S were 91.69% and 86.50% , respectively, and the mortality was 71.05% on the 14th day after inoculated with BbDp-06. The muscardine cadaver rate of larvae inoculated with M. anisopliae was significantly higher than that inoculated with ft bassiana. While LT50 value of the two M. anisopliae strains, Mal291-2 and Ma1775 were 5.73 and 5.39 days, respectively, and LT50 value of B. bassiana BbDp-06 was 8.02 days. LC50 value of two Btrains, Mal291-2 and BbDp-06 were 3.82 × 104 spore ·mL-1 and 3.88 × 104 spore·mL-1

  1. Virulence of Beauveria bassiana and Metarhizium anisopliae (Ascomycota: Hypocreales) commercial strains against adult Xylosandrus germanus (Coleoptera: Scolytidae) and impact on brood

    Science.gov (United States)

    The ambrosia beetle Xylosandrus germanus is an invasive pest with a wide host range and is a serious pest of orchards and nurseries in the eastern US. In this study we evaluated the potential of entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae as control agents against this beet...

  2. Altered immunity in crowded locust reduced fungal (Metarhizium anisopliae) pathogenesis.

    Science.gov (United States)

    Wang, Yundan; Yang, Pengcheng; Cui, Feng; Kang, Le

    2013-01-01

    The stress of living conditions, similar to infections, alters animal immunity. High population density is empirically considered to induce prophylactic immunity to reduce the infection risk, which was challenged by a model of low connectivity between infectious and susceptible individuals in crowded animals. The migratory locust, which exhibits polyphenism through gregarious and solitary phases in response to population density and displays different resistance to fungal biopesticide (Metarhizium anisopliae), was used to observe the prophylactic immunity of crowded animals. We applied an RNA-sequencing assay to investigate differential expression in fat body samples of gregarious and solitary locusts before and after infection. Solitary locusts devoted at least twice the number of genes for combating M. anisopliae infection than gregarious locusts. The transcription of immune molecules such as pattern recognition proteins, protease inhibitors, and anti-oxidation proteins, was increased in prophylactic immunity of gregarious locusts. The differentially expressed transcripts reducing gregarious locust susceptibility to M. anisopliae were confirmed at the transcriptional and translational level. Further investigation revealed that locust GNBP3 was susceptible to proteolysis while GNBP1, induced by M. anisopliae infection, resisted proteolysis. Silencing of gnbp3 by RNAi significantly shortened the life span of gregarious locusts but not solitary locusts. By contrast, gnbp1 silencing did not affect the life span of both gregarious and solitary locusts after M. anisopliae infection. Thus, the GNBP3-dependent immune responses were involved in the phenotypic resistance of gregarious locusts to fungal infection, but were redundant in solitary locusts. Our results indicated that gregarious locusts prophylactically activated upstream modulators of immune cascades rather than downstream effectors, preferring to quarantine rather than eliminate pathogens to conserve energy

  3. Altered immunity in crowded locust reduced fungal (Metarhizium anisopliae pathogenesis.

    Directory of Open Access Journals (Sweden)

    Yundan Wang

    2013-01-01

    Full Text Available The stress of living conditions, similar to infections, alters animal immunity. High population density is empirically considered to induce prophylactic immunity to reduce the infection risk, which was challenged by a model of low connectivity between infectious and susceptible individuals in crowded animals. The migratory locust, which exhibits polyphenism through gregarious and solitary phases in response to population density and displays different resistance to fungal biopesticide (Metarhizium anisopliae, was used to observe the prophylactic immunity of crowded animals. We applied an RNA-sequencing assay to investigate differential expression in fat body samples of gregarious and solitary locusts before and after infection. Solitary locusts devoted at least twice the number of genes for combating M. anisopliae infection than gregarious locusts. The transcription of immune molecules such as pattern recognition proteins, protease inhibitors, and anti-oxidation proteins, was increased in prophylactic immunity of gregarious locusts. The differentially expressed transcripts reducing gregarious locust susceptibility to M. anisopliae were confirmed at the transcriptional and translational level. Further investigation revealed that locust GNBP3 was susceptible to proteolysis while GNBP1, induced by M. anisopliae infection, resisted proteolysis. Silencing of gnbp3 by RNAi significantly shortened the life span of gregarious locusts but not solitary locusts. By contrast, gnbp1 silencing did not affect the life span of both gregarious and solitary locusts after M. anisopliae infection. Thus, the GNBP3-dependent immune responses were involved in the phenotypic resistance of gregarious locusts to fungal infection, but were redundant in solitary locusts. Our results indicated that gregarious locusts prophylactically activated upstream modulators of immune cascades rather than downstream effectors, preferring to quarantine rather than eliminate pathogens to

  4. Differential allergy responses to Metarhizium anisopliae fungal component extracts in BALB/c mice

    Science.gov (United States)

    Intratracheal aspiration (IA) exposure to Metarhizium anisopliae crude antigen (MACA), which is composed of equal protein amounts of mycelium (MYC), conidia (CON) and inducible proteases/chitinases (IND) extracts/filtrates, has resulted in responses characteristic of human allerg...

  5. Beauveria bassiana and Metarhizium anisopliae endophytically colonize cassava roots following soil drench inoculation

    Science.gov (United States)

    The fungal entomopathogens Beauveria bassiana and Metarhizium anisopliae were investigated to determine if endophytic colonization could be achieved in cassava. An inoculation method based on drenching the soil around cassava stems using conidial suspensions resulted in endophytic colonization of ca...

  6. Selectivity of Beauveria bassiana and Metarrhizium anisopliae to Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae); Seletividade de Beauveria bassiana e Metarhizium anisopliae a Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae)

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    Potrich, Michele; Silva, Everton L. da; Neves, Pedro M.O.J. [Universidade Estadual de Londrina (UEL), PR (Brazil). Centro de Ciencias Agrarias], e-mail: profmichele@gmail.com, e-mail: evertonloz@gmail.com, e-mail: pedroneves@uel.br; Alves, Luis F.A.; Daros, Alaxsandra [Universidade do Oeste do Parana (UNIOESTE), Cascavel, PR (Brazil). Centro de Ciencias Biologicas e da Saude. Lab. de Zoologia de Invertebrados], e-mail: lfaalves@unioeste.br; Haas, Jucelaine; Pietrowski, Vanda [Universidade do Oeste do Parana (UNIOESTE), Marechal Candido Rondon, PR (Brazil). Centro de Ciencias Agrarias], e-mail: jubarth@gmail.com, e-mail: vandapietrowski@gmail.com

    2009-07-01

    Trichogramma pretiosum Riley and the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae are efficient biological control agents and are thought to be used jointly. In here, we investigated if these entomopathogens could have any side-effects on T. pretiosum. Therefore, 1 x 8 cards containing sterilized eggs of Anagasta kuehniella (Zeller) that were sprayed with 0.2 ml of B. bassiana or M. anisopliae (1.0 X 10{sup 9} conidia/ml) were offered to a T. pretiosum female for 24h (30 cards/fungus = 30 replicates). Afterwards, females were isolated in glass tubes. The control group was sprayed with sterile distillated water + Tween 80 (0.01%). In addition, 60 cards with sterilized eggs of A. kuehniella were submitted to parasitism by females of T. pretiosum for 24h. Of these cards, 30 were sprayed with B. bassiana or M. anisopliae and 30 with distillated water + Tween 80 (0.01%), and observed daily until parasitoid emergence. Metarhizium anisopliae decreased parasitoid emergence and caused confirmed mortality. Therefore, field and semi-field experiments should be conducted for a final assessment of the side-effects of these entomopathogens on Trichogramma as a ways to develop a control strategy in which both can be used. (author)

  7. Susceptibility of the tick Haemaphysalis qinghaiensis to isolates of the fungus Metarhizium anisopliae in China.

    Science.gov (United States)

    Ren, Qiaoyun; Sun, Ming; Guan, Guiquan; Liu, Zhijie; Chen, Ze; Liu, Aihong; Li, Youquan; Ma, Miling; Yang, Jifei; Niu, Qingli; Liu, Junlong; Han, Xueqing; Yin, Hong; Luo, Jianxun

    2014-10-01

    Haemaphysalis qinghaiensis, a prevalent tick species in China, causes severe economic losses. In this study, we investigated the pathogenicity of six isolates of the fungus Metarhizium anisopliae to engorged female H. qinghaiensis using concentrations of 10(6), 10(7) and 10(8) conidia ml(-1). The results indicated that M.aAT08 and M.aAT13 isolates were highly virulent against the ticks. Metarhizium anisopliae has potential for biocontrol of H. qinghaiensis.

  8. Pathogenicity of entomopathogenic fungus Metarhizium anisopliae (Deuteromycetes) to Ixodes scapularis (Acari: Ixodidae)

    Science.gov (United States)

    Zhioua, E.; Browning, M.; Johnson, P.W.; Ginsberg, H.S.; LeBrun, R.A.

    1997-01-01

    The entomopathogenic fungus Metarhizium anisopliae is highly pathogenic to the black-legged tick, Ixodes scapularis. Spore concentrations of 108/ml for engorged larvae and 107/ml for engorged females resulted in 100% tick mortality, 2 wk post-infection. The LC50 value for engorged larvae (concentration to kill 50% of ticks) was 107 spores/ml. Metarhizium anisopliae shows considerable potential as a microbial control agent for the management of Ixodes scapularis.

  9. Sensitive and rapid detection of the insect pathogenic fungus Metarhizium anisopliae var. anisopliae by loop-mediated isothermal amplification.

    Science.gov (United States)

    Li, Ya; Cai, Shuang-Hu

    2011-05-01

    A set of six specific primers was designed by targeting intergenic spacer region (IGS) sequences. With Bst DNA polymerase, the products could be clearly amplified for 60 min at 62 °C in a simple water bath. The sensitivity of the loop-mediated isothermal amplification (LAMP) for detecting Metarhizium anisopliae var. anisopliae was about 0.01 pg fungal DNA per reaction (equivalent to 27 conidia). LAMP products could be judged with agar gel or naked eye after addition of SYBR Green I. There were no cross reactions with other fungal isolates indicating high specificity of the LAMP. The LAMP could detect the presence of M. anisopliae var. anisopliae from soil. The detection limits for M. anisopliae var. anisopliae of LAMP reaction was 50 conidia per reaction in soil.

  10. Laboratory and field evaluation of Metarhizium anisopliae var. anisopliae for controlling subterranean termites.

    Science.gov (United States)

    Hussain, A; Ahmed, S; Shahid, M

    2011-01-01

    The efficacy of the Metarhizium anisopliae strain ARSEF 6911 was determined in the laboratory and field against two sugarcane pests, Microtermes obesi Holmgren and Odontotermes obesus Rambur (Termitidae: Isoptera). The susceptibility of both termite species to different conidial suspensions (1 × 10(10), 1 × 10(8), 1 × 10(6) and 1 × 10(4) conidia/ml) was determined in laboratory. All conidial suspensions were able to induce mortality. Termite mortality caused by the fungal suspensions was dose dependent. There were no significant differences in the LT50 values between species. Field evaluation of M. anisopliae alone or in combination with diesel oil and thiamethoxam was carried out in two growing seasons (autumn 2005 and spring 2006) at two sites located in Punjab, Pakistan. Dipping the sugarcane setts in these suspensions was tried to determine their effects on germination and percentage of bud damage to sugarcane setts. All treatments significantly reduced termite infestation compared to the untreated control. The combined treatment of M. anisopliae and diesel oil significantly reduced insect damage by attaining higher germination > 55% and lower bud damage < 5.50% at both sites in both seasons. The results suggest that the application of M. anisopliae and diesel oil in combination might be a useful treatment option for the management of termites in sugarcane.

  11. Laboratory and field evaluation of Metarhizium anisopliae var. anisopliae for controlling subterranean termites

    Energy Technology Data Exchange (ETDEWEB)

    Hussain, A. [South China Agricultural University, Guangzhou (China). College of Natural Resources and Environment; Ahmed, S. [South China Agricultural University, Guangzhou (China). Dept. of Agricultural Entomology; Shahid, M., E-mail: solvia_aah@yahoo.co [University of Agriculture, Faisalabad (Pakistan). Dept. of Chemistry and Biochemistry

    2011-03-15

    The efficacy of the Metarhizium anisopliae strain ARSEF 6911 was determined in the laboratory and field against two sugarcane pests, Microtermes obesi Holmgren and Odontotermes obesus Rambur (Termitidae: Isoptera). The susceptibility of both termite species to different conidial suspensions (1 x 10{sup 10}, 1 x 10{sup 8}, 1 x 10{sup 6} and 1 x 10{sup 4} conidia/ml) was determined in laboratory. All conidial suspensions were able to induce mortality. Termite mortality caused by the fungal suspensions was dose dependent. There were no significant differences in the LT{sub 50} values between species. Field evaluation of M. anisopliae alone or in combination with diesel oil and thyamethoxam was carried out in two growing seasons (autumn 2005 and spring 2006) at two sites located in Punjab, Pakistan. Dipping the sugarcane setts in these suspensions was tried to determine their effects on germination and percentage of bud damage to sugarcane setts. All treatments significantly reduced termite infestation compared to the untreated control. The combined treatment of M. anisopliae and diesel oil significantly reduced insect damage by attaining higher germination > 55% and lower bud damage < 5.50% at both sites in both seasons. The results suggest that the application of M. anisopliae and diesel oil in combination might be a useful treatment option for the management of termites in sugarcane. (author)

  12. Horizontal Transmission of Beauveria bassiana (Hypocreales: Cordycipitaceae) and Metarhizium anisopliae (Hypocreales: Clavicipitaceae) in Musca domestica (Diptera: Muscidae).

    Science.gov (United States)

    Cárcamo, M C; Felchicher, F; Duarte, J P; Bernardi, E; Ribeiro, P B

    2015-08-01

    Beauveria bassiana Vuillemin and Metarhizium anisopliae (Metschnikoff) Sorokin are fungi with potential for controlling Musca domestica L. However, the impact on this dipteral may vary depending on the fungal isolates and the methodology used. This study evaluated the pathogenicity of direct application and horizontal transmission of B. bassiana (CG240) and M. anisopliae (CG34) on adult M. domestica individuals. The impact of B. bassiana and M. anisopliae on M. domestica was evaluated at the concentrations 2 × 10(4), 2 × 10(5), 2 × 10(6), and 2 × 10(7) conidia/ml. Horizontal transmission was also estimated between sexes at different infection periods of the vector insect. The mortality of adult M. domestica individuals directly infected with B. bassiana was above 90%, and the mortality of those infected with M. anisopliae ranged from 25.50 to 97.78%. Horizontal transmission of B. bassiana caused the death of 100% of individuals, in turn, that of M. anisopliae killed 55% of male and 100% of female individuals. Horizontal transmission of fungi was negatively influenced by time. This study shows the potential of these fungi for controlling M. domestica, both with the direct implementation strategy and horizontal transmission. However, field studies are needed to evaluate the capacity to decrease the M. domestica population using these alternatives.

  13. Diatomaceous earth and oil enhance effectiveness of Metarhizium anisopliae against Triatoma infestans.

    Science.gov (United States)

    Luz, Christian; Rodrigues, Juscelino; Rocha, Luiz F N

    2012-04-01

    Entomopathogenic fungi, especially Metarhizium anisopliae, have potential for integrated control of peridomestic triatomine bugs. However, the high susceptibility of these vectors to fungal infection at elevated ambient humidities decreases in the comparatively dry conditions that often prevail in their microhabitats. A formulation adapted to this target pest that induces high and quick mortality can help to overcome these drawbacks. In the present study diatomaceous earth, which is used against pests of stored grains or as an additive to mycoinsecticides, delayed but did not reduce in vitro germination of M. anisopliae s.l. IP 46 conidia after >24h agitation without affecting viability, and did not hamper the survival of Triatoma infestans nymphs exposed to treated surfaces. The settling behavior of nymphs on a treated surface in choice tests depended on the concentration of diatomaceous earth and ambient light level. Conidia formulated with diatomaceous earth and a vegetable oil synergized the insecticidal effect of the fungus in nymphs, and quickly killed all treated insects, even at 75% relative humidity (LT(90) 8.3 days) where unformulated conidia caused only 25% mortality after a 25 days exposure. The improved performance of a combined oil and desiccant dust formulation of this Metarhizium isolate raises the likelihood for its successful mycoinsecticidal use for triatomine control and, apparently, against other domestic insect pests.

  14. Production of Microsclerotia of Metarhizium anisopliae Using Deep-Tank Liquid Fermentation

    Science.gov (United States)

    The entomopathogenic fungus Metarhizium anisopliae is a pathogen of numerous soil-dwelling insects and has been registered in the United States and other countries as a bioinsecticide. Recent studies using various strains of M. anisopliae showed that small sclerotia (microsclerotia) were produced i...

  15. ASSESSMENT OF A CRUDE FUNGAL (METARHIZIUM ANISOPLIAE) EXTRACT AND IT'S COMPONENTS FOR ALLERGENICITY

    Science.gov (United States)

    ASSESSMENT OF A CRUDE FUNGAL (METARHIZIUM ANISOPLIAE) EXTRACT AND IT'S COMPONENTS FOR ALLERGENICITY. M D W Ward1, M E Viana2, L B Copeland1, and MJ K Selgrade1. 1US EPA, ORD, NHEERL, RTP, NC, USA. 2NCSU, College of Veterinary Medicine, Raleigh, NC, USA. Metarhizium anisopli...

  16. Application of the entomogenous fungus, Metarhizium anisopliae, for leafroller (Cnaphalocrocis medinalis) control and its effect on rice phyllosphere microbial diversity.

    Science.gov (United States)

    Hong, Mingsheng; Peng, Guoxiong; Keyhani, Nemat O; Xia, Yuxian

    2017-07-10

    Microbial pesticides form critical components of integrated pest management (IPM) practices. Little, however, is known regarding the impacts of these organisms on the indigenous microbial community. We show that Metarhizium anisopliae strain CQMa421 was highly effective in controlling the rice leafroller, Cnaphalocrocis medinalis Guenee. In addition, M. anisopliae distribution and its effects on phyllosphere microbial diversity after application in field trials were investigated. Phylloplane specific distribution of the fungus was observed over time, with more rapid declines of M. anisopliae CFUs (colony-forming units) seen in the top leaf layer as compared to lower layers. Application of the fungus resulted in transient changes in the endogenous microbial diversity with variations seen in the bacterial observed species and Shannon index. Notable increases in both parameters were seen at 6-day post-application of M. anisopliae, although significant variation within sample replicates for bacteria and fungi were noted. Application of M. anisopliae increased the relative distribution of bacterial species implicated in plant growth promotion and organic pollutant degradation, e.g., Methylobacterium, Sphingobium, and Deinococcus. These data show minimal impact of M. anisopliae on endogenous microbial diversity with transient changes in bacterial abundance/diversity that may result in added benefits to crops.

  17. Enzymatic activities and effects of mycovirus infection on the virulence of Metarhizium anisopliae in Rhipicephalus microplus.

    Science.gov (United States)

    Perinotto, Wendell M S; Golo, Patricia S; Coutinho Rodrigues, Caio J B; Sá, Fillipe A; Santi, Lucélia; Beys da Silva, Walter O; Junges, Angela; Vainstein, Marilene H; Schrank, Augusto; Salles, Cristiane M C; Bittencourt, Vânia R E P

    2014-06-16

    The present study aimed to evaluate the pathogenic potential of different Metarhizium anisopliae s.l. isolates and to determine whether differences in enzymatic activities of proteases, lipases and chitinases and infection with mycoviruses affect the control of Rhipicephalus microplus achieved by these fungal isolates. Engorged female ticks were exposed to fungal suspensions. The lipolytic and proteolytic activities in the isolates were evaluated using chromogenic substrates and the chitinolytic activity was determined using fluorescent substrates. A gel zymography was performed to determine the approximate size of serine proteases released by M. anisopliae isolates. To detect mycoviral infections, dsRNA was digested using both RNAse A and S1 endonuclease; samples were analyzed on an agarose gel. Four of the five isolates tested were infected with mycovirus; however, the level of control of R. microplus ticks achieved with the only isolate free of infection (isolate CG 347) was low. This finding suggests that mycoviral infection does not affect the virulence of fungi against ticks. Although all five isolates were considered pathogenic to R. microplus, the best tick control and the highest levels of enzymatic activity were achieved with the isolates CG 629 and CG 148. The in vitro activities of lipases, proteases and chitinases produced by M. anisopliae s.l. differed among isolates and may be related to their virulence.

  18. PERSISTÊNCIA DE Metarhizium anisopliae spp NO SOLO SOB DIFERENTES CONDIÇÕES DE TEMPERATURA E UMIDADE

    Directory of Open Access Journals (Sweden)

    Diana Mendonça Silva Guerra

    2009-01-01

    Full Text Available The fungi entomopathogenic are actually objects of works according to their importance in the ecological system. This work analysed the persistence of Metarhizium anisopliae var. anisopliae and M. anisopliae var. acridum, under different conditions of temperature and humidity, in the period of September to December. After inoculation on soil the fungi were submitted to four different treatments: environment temperature and 25% of humidity; environment temperature and 75% of humidity; 28º C and 25% of humidity; 28º C and 75% of humidity, during 120 days. The results show that M. anisiopliac var. anisopliae presented the most recuperation in the treatment at 28º C and 75% of humidity (P > 0,05 in 30 days and maintained a positive conditions on soil for 120 days of experiment. The same did not happen to M. anisopliae var. acridum that, during the whole process of observation and controll the colonies recuperation avoiding its persistence on soil, during the 120 days of experiment.

  19. Characterization of a Peruvian isolate of Metarhizium anisopliae var. acridum, a pathogen of grasshoppers

    Directory of Open Access Journals (Sweden)

    Magalhães Bonifácio Peixoto

    2003-01-01

    Full Text Available The objective of this study was to characterize the Peruvian isolate of Metarhizium anisopliae var. acridum, CG 863, obtained from the grasshopper Schistocerca interrita, a crop pest in Peru. The characterization was done by comparing this isolate with two other ones of M. anisopliae var. acridum, from Brazil and Australia, and with an isolate of M. anisopliae var. anisopliae. The three M. anisopliae var. acridum isolates had similar growth profiles in agar plates at 25°C and 37°C, and similar RAPD patterns according to the analysis of three primers. However, regarding these parameters and conidial size, these isolates were very distinct when compared to M. anisopliae var. anisopliae isolate. Bioassays indicated that the Peruvian isolate is as pathogenic as the Brazilian isolate against nymphs of Rhammatocerus schistocercoides.

  20. Patogenicidade e viabilidade de Beauveria bassiana, Metarhizium anisopliae Var. anisopliae e Metarhizium anisopliae Var. acridium ao Anthonomus grandis (Boheman) (Coleoptera : Curculionidae)

    OpenAIRE

    2005-01-01

    Fungos entomopatogênicos Beauveria bassiana e Metarhizium anisopliae amplamente distribuídos em todas as regiões do mundo podem ser isolados de insetos, solo e de outros substratos. Essas espécies apresentam variabilidade genética e habilidades sobre diferentes insetos-praga. Patogenicidade e virulência sobre diversos artrópodes têm sido observadas com a variação de atividades enzimáticas e características genéticas dos isolados. A ação desses agentes está relacionada com as condições climáti...

  1. Infection of Psoroptes mites with the fungus Metarhizium anisopliae.

    Science.gov (United States)

    Brooks, A J; Wall, R

    2001-01-01

    The astigmatid mite, Psoroptes ovis (Hering) (Acari: Psoroptidae), is an obligatory ectoparasite that causes psoroptic mange in a range of domesticated animals, particularly sheep, where the clinical disease is known as sheep scab. A series of laboratory assays were used to assess the use of the fungus, Metarhizium anisopliae (Metchnikoff) (Deuteromycotina: Hyphomycetes) as a biocontrol agent for P. ovis derived from rabbits (syn. P. cuniculi). The immersion of mites in a suspension of conidia of M. anisopliae resulted in the acquisition of fatal infections. The number of mites which developed infections increased significantly with the increasing concentration of the conidial suspension to which they were exposed; 77% of mites developed infections when exposed to the highest concentration used (1 x 10(8) conidia ml(-1)). Controls developed no fungal infections. Mites allowed simply to walk across a surface which had been treated with a suspension of conidia also acquired fungal infections; the number infected was again related to the concentration of conidia present. After contact for 24 h with a surface treated with 1 x 10(8) conidia ml(-1), 73% of the mites became infected. To determine whether dead infected mites could act as sources of infection, infected cadavers were placed in chambers with live uninfected mites. The uninfected mites acquired fatal infections from the cadavers; a higher ratio of infected cadavers to uninfected mites resulted in greater transmission of infection. The time after death of the infected cadaver was also an important factor influencing the number infected, 5-day-old cadavers were the most infective and 18-day-old cadavers the least infective. The results indicate that M. anisopliae is a good candidate control agent for Psoroptes mites.

  2. Laboratory evaluation of three strains of the entomopathogenic fungus Metarhizium anisopliae for controlling Dermanyssus gallinae.

    Science.gov (United States)

    Tavassoli, M; Ownag, A; Pourseyed, S H; Mardani, K

    2008-06-01

    The pathogenicity of three strains of the entomopathogenic fungus Metarhizium anisopliae on different life stages of Dermanyssus gallinae was evaluated in the laboratory. All the strains tested were virulent to D. gallinae but pathogenicity varied among the strains. Strain V245 induced a higher mortality rate using different concentrations than other two strains. The estimated median lethal concentration of different strains of M. anisopliae against D. gallinae varied depending on the exposure time of D. gallinae to M. anisopliae. It was concluded that the pathogenicity of the entomopathogenic fungus M. anisopliae on different life stages of D. gallinae was concentration and time dependent.

  3. [Bahaviour of Solenopsis invicta workers to protect pupae from infection by Metarhizium anisopliae].

    Science.gov (United States)

    Qiu, Hua-Long; Lü, Li-Hua; Zhang, Chun-Yang; He, Yu-Rong

    2014-09-01

    Previous studies have focused on how ants deal with workers infected by pathogens but how pupae are protected from infection by fungi is not well understood. The behavioral mechanisms adopted by Solenopsis invicta (red imported fire ants, RIFA) adult workers to protect pupae against Metarhizium anisopliae infection were studied. We observed the behavioral changes of M. anisopliae infected adult workers in the brood chamber as well as the behavioral changes of healthy workers to fungus exposed pupae. The time of fungus infected workers spent in the pupal chamber reduced significantly from 103.4 s on the first day to 38.5 s on the third day. Moreover, the percentage of time spending on brood care in the pupal chamber reduced significantly from 13.6% on the first day to 3.5% on the third day. When pupae were infected by M. anisopliae, workers performed 5.3 times more grooming to fungus exposed pupae than controls, and the duration of each grooming bout to fungus exposed pupae was 5.2 times longer than controls. Grooming did remove many conidia on the surface of fungus exposed pupae. The mean numbers of conidia on the surface of pupae were 103.1, 51.6 and 31.3 when no workers, two workers and ten workers accompanied a pupa, respectively. The presence of workers resulted in a lower germination rate of conidia on the surface of pupae. The mean germination rates of conidia after 20 h of inoculation on the surface of pupae were 95.1%, 80.4% and 59.9%, in the treatments with no worker, two workers and ten workers respectively. There was a positive correlation between the emergence rate of pupae and the number of accompanying workers. RIFA protect their pupae from infection by M. anisopliae through social be- haviors which enable the sustainable development of their population.

  4. Survival and immune response of the Chagas vector Meccus pallidipennis (Hemiptera: Reduviidae) against two entomopathogenic fungi, Metarhizium anisopliae and Isaria fumosorosea

    OpenAIRE

    Flores-Villegas, A. Laura; Cabrera-Bravo,Margarita; Toriello, Conchita; Bucio-Torres, Martha I.; Salazar-Schettino,Paz María; Córdoba-Aguilar, Alex

    2016-01-01

    Background Chagas disease is a key health problem in Latin America and is caused and transmitted by Trypanosoma cruzi and triatomine bugs, respectively. Control of triatomines has largely relied on the use pyrethroids, which has proved to be ineffective in the long term. Alternatively, the use of entomopathogenic fungi has been implemented to control triatomine bugs. These fungi are highly efficient as they induce a reduction in immune response on insects. Meccus pallidipennis is the main tri...

  5. Virulence potential of Metarhizium anisopliae s.l. isolates on Rhipicephalus (Boophilus) microplus larvae.

    Science.gov (United States)

    Quinelato, Simone; Golo, Patrícia S; Perinotto, Wendell M S; Sá, Fillipe A; Camargo, Mariana G; Angelo, Isabele C; Moraes, Aurea M L; Bittencourt, Vânia R E P

    2012-12-21

    The use of entomopathogenic fungi to control arthropods has been reported worldwide for decades. The aim of the present study was to evaluate the virulence of 30 Metarhizium anisopliae s.l. Brazilian isolates from different geographical regions, hosts or substrates on the larvae of Rhipicephalus (Boophilus) microplus ticks under in vitro conditions to the selection of virulent isolates in order to be further used in biological control programs. The current study confirmed the lethal action of M. anisopliae s.l. isolates on R. (B.) microplus larvae with different mortality levels, usually directly proportional to the conidia concentration. No relationship was found between the origin of the isolate and its virulence potential or between the virulence potential and conidia production. Three isolates (CG 37, CG 384 and IBCB 481) caused a high percentage of larval mortality, reaching LC(50) at 10(6) conidia ml(-1), thus requiring a lower conidia concentration to cause an approximately 100% larval mortality. The results of this study suggest that these three isolates are the most promising for use in programs aimed at microbial control in the field.

  6. Use of Metarhizium anisopliae chitinase genes for genotyping and virulence characterization.

    Science.gov (United States)

    Niassy, Saliou; Subramanian, Sevgan; Ekesi, Sunday; Bargul, Joel L; Villinger, Jandouwe; Maniania, Nguya K

    2013-01-01

    Virulence is the primary factor used for selection of entomopathogenic fungi (EPF) for development as biopesticides. To understand the genetic mechanisms underlying differences in virulence of fungal isolates on various arthropod pests, we compared the chitinase genes, chi2 and chi4, of 8 isolates of Metarhizium anisopliae. The clustering of the isolates showed various groups depending on their virulence. However, the analysis of their chitinase DNA sequences chi2 and chi4 did not reveal major divergences. Although their protein translates have been implicated in fungal virulence, the predicted protein structure of chi2 was identical for all isolates. Despite the critical role of chitin digestion in fungal infection, we conclude that chi2 and chi4 genes cannot serve as molecular markers to characterize observed variations in virulence among M. anisopliae isolates as previously suggested. Nevertheless, processes controlling the efficient upregulation of chitinase expression might be responsible for different virulence characteristics. Further studies using comparative "in vitro" chitin digestion techniques would be more appropriate to compare the quality and the quantity of chitinase production between fungal isolates.

  7. Use of Metarhizium anisopliae Chitinase Genes for Genotyping and Virulence Characterization

    Directory of Open Access Journals (Sweden)

    Saliou Niassy

    2013-01-01

    Full Text Available Virulence is the primary factor used for selection of entomopathogenic fungi (EPF for development as biopesticides. To understand the genetic mechanisms underlying differences in virulence of fungal isolates on various arthropod pests, we compared the chitinase genes, chi2 and chi4, of 8 isolates of Metarhizium anisopliae. The clustering of the isolates showed various groups depending on their virulence. However, the analysis of their chitinase DNA sequences chi2 and chi4 did not reveal major divergences. Although their protein translates have been implicated in fungal virulence, the predicted protein structure of chi2 was identical for all isolates. Despite the critical role of chitin digestion in fungal infection, we conclude that chi2 and chi4 genes cannot serve as molecular markers to characterize observed variations in virulence among M. anisopliae isolates as previously suggested. Nevertheless, processes controlling the efficient upregulation of chitinase expression might be responsible for different virulence characteristics. Further studies using comparative “in vitro” chitin digestion techniques would be more appropriate to compare the quality and the quantity of chitinase production between fungal isolates.

  8. Variation in the electrophoretic karyotype of Brazilian strains of Metarhizium anisopliae

    Directory of Open Access Journals (Sweden)

    Valadares-Inglis Maria Cléria

    1998-01-01

    Full Text Available Pulsed-field gel electrophoresis (PFGE was used to separate chromosome-sized DNA molecules of four strains of Metarhizium anisopliae from Brazil. Metarhizium anisopliae isolates from Japan have been reported as possessing seven chromosomes. Variation was observed among the Brazilian strains and the chromosomal DNA was resolved into eight bands for strain CG46. Densitometric analysis of PFGE gels suggested that the other three Brazilian strains also possess eight chromosomes, with two chromosomes migrating as doublets under the electrophoretic conditions used. The genome size was estimated as varying between 23.39 to 31.88 Mb, not including possible doublet chromosomes.

  9. Genome-wide identification and profiling of microRNA-like RNAs from Metarhizium anisopliae during development.

    Science.gov (United States)

    Zhou, Quan; Wang, Zhangxun; Zhang, Jun; Meng, Huimin; Huang, Bo

    2012-11-01

    Metarhizium anisopliae is one of the most common species of entopathogenic fungi. It has economic and social benefits in many countries where used in agriculture as an important biological control agent of insect pests. M. anisopliae can exist as multiple cell types, which suggests that this fungus has a complex way of gene regulation. MicroRNAs (miRNAs) are endogenous small noncoding RNAs. They play a crucial role in the regulation of gene expression and cell function in plants, animals, and in fungi where they were termed miRNA-like RNAs (milRNAs). In this study, we aimed to identify potential milRNAs in M. anisopliae that may regulate the processes of mycelium growth and conidiogenesis (CO). Two small RNA (sRNA) libraries were constructed and submitted to Solexa sequencing. Fifteen milRNAs were identified using deep-sequencing and computational analysis; most of these milRNAs originated from single genes. Database searches revealed that these novel milRNAs had no homologues in other organisms and were, therefore, M. anisopliae-specific. Many of the milRNAs had differential expression profiles for either mycelium growth or CO. The expression of the selected milRNAs was validated by quantitative reverse transcription polymerase chain reaction. Seventy-eight potential target mRNAs for 14 of the milRNAs were identified successfully by computational analysis. These milRNAs may play an important role in the regulation of mycelial growth and conidiation in M. anisopliae. To our knowledge, this study is the first report of milRNA profiles of organisms in the order Hypocreales. This information could be used to study the regulation of genes and their networks in M. anisopliae.

  10. Virulence and Pre-Lethal Reproductive Effects of Metarhizium anisopliae var. anisopliae on Pseudococcus viburni (Hemiptera: Pseudococcidae Virulencia y Efectos Pre-Letales en la Reproducción de Metarhizium anisopliae var. anisopliae en Pseudococcus viburni (Hemiptera: Pseudococcidae

    Directory of Open Access Journals (Sweden)

    Aldo Pereira

    2011-12-01

    Full Text Available Obscure mealybug, Pseudococcus viburni (Signoret, is a cosmopolitan pest that causes damage by suction of vascular juices and the production of honeydew, as well as for being a quarantine insect. Within control options, entomopathogenic fungi are a good alternative, nevertheless, more research is needed. In this research, the Metarhizium anisopliae var. anisopliae (Metschnikoff isolate Qu-M984 was evaluated on P. viburni under laboratory conditions. Virulence was evaluated by lethal dose 50 (LD50 and lethal time 50 (LT50, for each of the four life stages of the female. The doses tested were 10(5, 10(6, 10(7 and 10(8 conidia mL-1. Also fecundity, egg size, fertility, and longevity of adult females were evaluated at doses of 10(5 and 10(6 conidia mL-1, the evaluations were made every 2 d throughout the insect life time. The LD50 and LT50 obtained were variable for each life stage, although without statistical differences among life stages (P > 0.05, ranging from LD50 = 7.3 x 10(5 to 4.9 x 10(9 conidia mL-1 and LT50 = 7.74 to 9.97 d at 10(8 conidia mL-1. Significant differences (P 0.05.Chanchito blanco de la vid, Pseudococcus viburni (Signoret, es una plaga cosmopolita que causa daños tanto por succión de jugos vasculares como por su producción de mielecilla, así como también por ser un insecto cuarentenario. Dentro de las opciones de control, hongos entomopatógenos son una buena alternativa, sin embargo, más investigación es necesaria. En esta investigación fue evaluado Metarhizium anisopliae var. anisopliae (Metschnikoff aislamiento Qu-M984 en P. viburni bajo condiciones de laboratorio. Fue evaluada virulencia según dosis letal 50 (LD50 y tiempo letal 50 (LT50 para cada uno de los cuatro estados de desarrollo de la hembra. Las dosis evaluadas fueron 10(5, 10(6, 10(7 y 10(8 conidias mL-1. Fecundidad, tamaño de huevos, fertilidad y longevidad de hembras adultas fueron evaluados con dosis 10(5 y 10(6 conidias mL-1, las evaluaciones fueron

  11. Application of Bait Treated with the Entomopathogenic Fungus Metarhizium anisopliae (Metsch. Sorokin for the Control of Microcerotermes diversus Silv.

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    Amir Cheraghi

    2013-01-01

    Full Text Available Microcerotermes diversus Silvestri (Isoptera, Termitidae is considered to be the most destructive termite in Khuzestan province (Iran, and its control by conventional methods is often difficult. Biological control using entomopathogenic fungi could be an alternative management strategy. Performance of a bait matrix treated with the entomopathogenic fungus Metarhizium anisopliae (Metsch. Sorokin, Strain Saravan (DEMI 001, against M. diversus was evaluated in this paper. The highest rate of mortality occurred at concentrations of 3.7 × 107 and 3.5 × 108 (conidia per mL. There was no significant difference between treatments, in the rate of feeding on the bait. The fungal pathogen was not repellent to the target termite over the conidial concentrations used. The current results suggest potential of such bait system in controlling termite. However the effectiveness of M. anisopliae as a component of integrated pest management for M. diversus still needs to be proven under field conditions.

  12. Evaluating the virulence of Metarhizium anisopliae (Deuteromycotina: Hyphomycetes and Beauveria bassiana (Ascomycota: Hypocreales isolates to Arabian rhinoceros beetle, Oryctes agamemnon arabicus

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    M.W. Khudhair

    2015-12-01

    Full Text Available Virulence of entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana were tested against Arabian Rhinoceros Beetle, Oryctes agamemnon arabicus larvae. Four concentrations (1×105, 1×107, 1×109 and 1×1011 conidia/mL–1 of two locally isolated entomopathogenic fungi spore suspensions were used in this study via larval direct spraying. Results revealed that both isolates can cause high mortality rate reaching 100% after 29 days. However, Beauveria bassiana scored higher mortality rate in short time especially at the concentration of 1×1011 conidia/ mL–1 with lethal time (LT50 12.75 and LT90 20.00; while, Metarhizium anisopliae caused the higher percentage of malformed adults. Moreover, both isolates affected insect’s life cycle particularly in the pupal stage which was reduced remarkably by almost 50% in comparison with the control treatment.

  13. Patogenicidade de Metarhizium anisopliae var. anisopliae, Metarhizium flavoviride var. flavoviride e Beauveria bassiana a Nasutitermes coxipoensis (Isoptera: Termitidae) em cana-de-açúcar e compatibilidade com inseticidas químicos

    OpenAIRE

    2005-01-01

    Metarhizium anisopliae var. anisopliae, Metarhizium flavoviride var. flavoviride e Beauveria bassiana têm ação comprovada contra diversas espécies de insetos. Os cupins podem ser infectados por esses fungos naturalmente. Avaliou-se a ação desses fungos sobre Nasutitermes coxipoensis em laboratório e quantificou-se a produção de conídios nos insetos mortos. A compatibilidade de M. anisopliae var. anisopliae reisolado de N. coxipoensis com os inseticidas fipronil, imidaclopride e ensossulfan fo...

  14. Pathogenicity of Metarhizium anisopliae (Deuteromycetes) and permethrin to Ixodes scapularis (Acari: Ixodidae) nymphs

    Science.gov (United States)

    Hornbostel, V.L.; Zhioua, Elyes; Benjamin, Michael A.; Ginsberg, Howard S.; Ostfeld, Richard S.

    2005-01-01

    Effectiveness of the entomopathogenic fungus Metarhizium anisopliae, for controlling nymphal Ixodes scapularis, was tested in laboratory and field trials. In the laboratory, M. anisopliae (Metschnikoff) Sorokin strain ESC1 was moderately pathogenic, with an LC50 of 107 spores/ml and induced 70% mortality at 109 spores/ml. In a field study, however, 109 spores/ml M. anisopliae did not effectively control questing I. scapularis nymphs, and significant differences were not detected in pre- and post-treatment densities. For nymphs collected and returned to the laboratory for observation, mortality was low in treatment groups, ranging from 20 to 36%. To assess whether a chemical acaricide would synergistically enhance pathogenicity of the fungus, we challenged unfed nymphal I. scapularis with combinations of M. anisopliae and permethrin, a relatively safe pyrethroid acaricide, in two separate bioassays. Significant interactions between M. anisopliae and permethrin were not observed, supporting neither synergism nor antagonism.

  15. Soil Persistence of Metarhizium anisopliae Applied to Manage Sugarbeet Root Maggot in a Cover Crop Microenvironment

    Science.gov (United States)

    The sugarbeet root maggot, Tetanops myopaeformis (Röder), is a major insect pest of sugarbeet, Beta vulgaris L., in North Dakota, Minnesota, and Idaho. Three field trials using the insect pathogen Metarhizium anisopliae (Metch.) Sorok. ATCC 62176 in conjunction with cover crops were conducted in 200...

  16. THE PESTICIDE METARHIZIUM ANISOPLIAE HAS AN ADJUVANT EFFECT ON THE ALLERGIC RESPONSE TO OVALBUMIN IN MICE

    Science.gov (United States)

    Metarhizium anisopliae is a parasitic fungus employed as a biological control agent against vermin and used in the US for indoor control of cockroaches. Sensitization to cockroach allergens is associated with development of asthma. This pesticide is non-pathogenic for humans and ...

  17. THE IDENTIFICATION AND CHARACTERIZATION OF AN IGE-INDUCING PROTEIN IN METARHIZIUM ANISOPLIAE EXTRACT

    Science.gov (United States)

    The Identification and Characterization of an IgE-Inducing Protein in Metarhizium anisopliae ExtractMarsha D.W. Ward1, Lisa B. Copeland1, Maura J. Donahue2, and Jody A. Shoemaker31ORD, NHEERL, US EPA, RTP, NC; 2Oak Ridge Institute for Science and Education, Cincinnati...

  18. Infection of adult Aedes aegypti and Ae. albopictus mosquitoes with the entomopathogenic fungus Metarhizium anisopliae

    NARCIS (Netherlands)

    Scholte, E.J.; Takken, W.; Knols, B.G.J.

    2007-01-01

    This study describes a laboratory investigation on the use of the insect-pathogenic fungus Metarhizium anisopliae against adult Aedes aegypti and Ae. albopictus mosquitoes. At a dosage of 1.6 × 1010 conidia/m2, applied on material that served as a mosquito resting site, an average of 87.1 ± 2.65% of

  19. Infection of adult Aedes aegypti and Ae. albopictus mosquitoes with the entomopathogenic fungus Metarhizium anisopliae

    NARCIS (Netherlands)

    Scholte, E.J.; Takken, W.; Knols, B.G.J.

    2007-01-01

    This study describes a laboratory investigation on the use of the insect-pathogenic fungus Metarhizium anisopliae against adult Aedes aegypti and Ae. albopictus mosquitoes. At a dosage of 1.6 × 1010 conidia/m2, applied on material that served as a mosquito resting site, an average of 87.1 ± 2.65% of

  20. Survival of anopheline eggs and their susceptibility to infection with Metarhizium anisopliae and Beauveria bassiana under laboratory conditions

    NARCIS (Netherlands)

    Luz, C.; Mnyone, L.L.; Russell, T.L.

    2011-01-01

    The viability of Anopheles gambiae sensu stricto and Anopheles arabiensis (Diptera: Culicidae) eggs over time and the ovicidal activity of Beauveria bassiana (Ascomycota: Cordycipitaceae) and Metarhizium anisopliae (Ascomycota: Clavicipitaceae) were investigated. Eggs were incubated in soil or leaf

  1. Evaluation of Metarhizium anisopliae, Beauveria bassiana and Paecilomyces fumosoroseus as entomopathogens of the cactus moth, Cactoblastis cactorum (Lepidoptera:Pyralidae)

    Science.gov (United States)

    The fungal pathogens Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae), Paecilomyces fumosoroseus (Wize) Brown & Smith (Deuteromycotina: Hyphomycetes), and Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Clavicipitaceae) were evaluated as potential biological control ...

  2. Differential expression of the pr1A gene in Metarhizium anisopliae and Metarhizium acridum across different culture conditions and during pathogenesis

    Directory of Open Access Journals (Sweden)

    Mariele Porto Carneiro Leão

    2015-03-01

    Full Text Available The entomopathogenic fungi of the genus Metarhizium have several subtilisin-like proteases that are involved in pathogenesis and these have been used to investigate genes that are differentially expressed in response to different growth conditions. The identification and characterization of these proteases can provide insight into how the fungus is capable of infecting a wide variety of insects and adapt to different substrates. In addition, the pr1A gene has been used for the genetic improvement of strains used in pest control. In this study we used quantitative RT-PCR to assess the relative expression levels of the pr1A gene in M. anisopliae and M. acridum during growth in different culture conditions and during infection of the sugar cane borer, Diatraea saccharalis Fabricius. We also carried out a pathogenicity test to assess the virulence of both species against D. saccharalis and correlated the results with the pattern of pr1A gene expression. This analysis revealed that, in both species, the pr1A gene was differentially expressed under the growth conditions studied and during the pathogenic process. M. anisopliae showed higher expression of pr1A in all conditions examined, when compared to M. acridum. Furthermore, M. anisopliae showed a greater potential to control D. saccharalis. Taken together, our results suggest that these species have developed different strategies to adapt to different growing conditions.

  3. EFEITO ANTAGÔNICO DE Trichoderma sp. NO DESENVOLVIMENTO DE Beauveria bassiana (Bals. Vuill. e Metarhizium anisopliae (Metsch. Sorok ANTAGONISTIC EFFECT OF Trichoderma sp. ON THE DEVELOPMENT OF Beauveria bassiana (Bals. Vuill. AND Metarhizium anisopliae (Metsch. SOROK

    Directory of Open Access Journals (Sweden)

    Alcides Moino Jr.

    1999-01-01

    Full Text Available Este trabalho teve por objetivo avaliar o efeito de Trichoderma sp. no desenvolvimento de Beauveria bassiana e Metarhizium anisopliae. Trichoderma sp., B. bassiana (isolado 634 e M. anisopliae (isolado E-9 foram inoculados em meio BDA, com intervalos de 0, 48, 120 e 168 horas entre a inoculação de Trichoderma sp. e dos entomopatógenos. Avaliou-se o crescimento radial das colônias nos períodos de 48 e 120 horas após a inoculação de Trichoderma sp., sendo que este afetou o desenvolvimento dos entomopatógenos quando inoculado simultaneamente ou após 48 horas. B. bassiana e M. anisopliae desenvolveram-se normalmente quando inoculados 168 horas antes de Trichoderma sp.. Também foi avaliado o efeito de um extrato de Trichoderma sp. sobre os entomopatógenos, com a adição de 0,1; 0,5; 1,0 e 5,0 ml de extrato/100,0 ml de meio, onde foram inoculados os entomopatógenos. Foram medidos os diâmetros de colônias e o número de conídios produzidos por B. bassiana e M. anisopliae na presença do extrato. A concentração de 5,0 ml de extrato/100,0 ml de meio alterou o crescimento e a conidiogênese de B. bassiana. O fungo M. anisopliae foi afetado a partir da adição de 1,0 ml de extrato/100,0 ml de meio.The objective of this work was to evaluate the effect of Trichoderma sp. on the development of Beauveria bassiana and Metarhizium anisopliae. The fungus Trichoderma sp. was inoculated on PDA culture medium, 0, 48, 120 and 168 hours after inoculation of the same plates with either B. bassiana (isolate 634 or M. anisopliae (isolate E-9. The radial growth of fungal colonies was measured 48 and 120 hours after Trichoderma sp. inoculation. Trichoderma sp. affected the development of both entomopathogenic fungi when inoculated simultaneously or 48 hours later. B. bassiana and M. anisopliae had normal development when inoculated 168 hours before Trichoderma sp. The effect of a toxic extract from Trichoderma sp. on the entomopathogenic fungi was also

  4. Effects of Fungicides on the Development of the Entomopathogenic Fungus Metarhizium anisopliae var. anisopliae Efecto de los Fungicidas sobre el Desarrollo del Hongo Entomopatógeno Metarhizium anisopliae var. anisopliae

    Directory of Open Access Journals (Sweden)

    Maribel Yáñez

    2010-09-01

    Full Text Available Metarhizium anisopliae (Metschnikoff Sorokin is an entomopathogenic fungus used for controlling different insect pests. It is most frequently applied to berry fruit crops, where fungicides are also used for disease control. Fungicides: azoxystrobin, benomyl, captan, chlorothalonil, fenhexamid, fludioxonil, iprodione, and metalaxyl in concentrations of 0.01, 0.1, 1.0, 10, and 100 mg L-1 were evaluated in this research study. Vegetative growth, conidia germination, and conidia germination tube length were measured on the Qu-M82, Qu-M151b, Qu-M253, Qu-M430, and Qu-M984 Metarhizium anisopliae var. anisopliae fungus strains. Those strains were selected because of their present use against different insect pest in bramble fruits. Vegetative growth was measured through the colony rate growth in agar media, and those reaching up to 50% of the check growth were considered compatible. Results indicate that the benomyl and fenhexamid fungicides were compatible with the five isolates whereas, azoxystrobin and fludioxonil were incompatible. Furthermore, benomyl and fludioxonil reduced conidia germination by 53 and 91%, and germination tube length by 18 and 37%, respectively.Metarhizium anisopliae (Metschnikoff Sorokin es un hongo entomopatógeno que se utiliza para el control de diferentes insectos, uno de sus usos más frecuentes es en frutales menores, donde también se utilizan fungicidas para el control de enfermedades. En este trabajo se evaluó el efecto de los fungicidas azoxystrobin, benomil, captan, chlorothalonil, fenhexamid, fludioxonil, iprodione y metalaxil, en concentraciones de 0,01; 0,1; 1; 10 y 100 mg L-1, sobre el crecimiento de la colonia, porcentaje de germinación de conidias y longitud de tubos germinativos de distintas cepas de M. anisopliae var. anisopliae. Las cepas utilizadas fueron Qu-M82, Qu-M151b, Qu-M253, Qu-M430 y Qu-M984, seleccionadas por su uso comercial para el control de diferentes insectos en frutales menores. El

  5. Selection of Beauveria bassiana (Bals. Vuill. and Metarhizium anisopliae (Metsch. Sorok. strains for control of Cornitermes cumulans (Kollar

    Directory of Open Access Journals (Sweden)

    Pedro J. Neves

    2000-01-01

    Full Text Available Fifty strains of the entomopathogenic fungi Beauveria bassiana (Bals. Vuill. and Metarhizium anisopliae (Metsch. Sorok. were tested against the termite Cornitermes cumulans. In the first phase of the experiments, several bioassays were conducted and the five best strains were selected. The criterion for strain selection during this phase was the confirmed mortality above 50% five days after application of the fungus. Three M. anisopliae and two B. bassiana strains were the most virulent. The second phase of the experiments consisted of a bioassay and a conidial production test using a rice medium. The best M. anisopliae and B. bassiana strains were selected using both the confirmed insect mortality nine days after application of the fungus and the yield of conidia. Considering results from the bioassay, M. anisopliae 1037 showed the highest confirmed (57.8% and total (89.2% mortalities after nine days. Among the B. bassiana strains, 447 was the most virulent with confirmed and total mortalities of 45.9% and 89.8%, respectively. M. anisopliae 1037 had the highest conidial yield with mean of 3.37 x 10(12 conidia/kg of rice, followed by B. bassiana strain 447 with 2.66 x 10(12 conidia/kg of rice. The M. anisopliae strain 1037 was the highest virulent followed by B. bassiana strain 447. This strains showed the greatest potential as agents to be used in biological control programs against C. cumulans.Nesta pesquisa foram testados cinqüenta isolados dos fungos entomopatogênicos Beauveria bassiana (Bals. Vuill. e Metarhizium anisopliae (Metsch. Sorok. sobre Cornitermes cumulans. Na primeira fase foram realizados vários bioensaios, selecionando-se os 5 melhores isolados. O critério usado para a seleção foi a mortalidade confirmada acima de 50%, no quinto dia após a aplicação. Três isolados de M. anisopliae e dois de B. bassiana foram os mais virulentos. A segunda fase de seleção constou de um bioensaio e um teste de produção de con

  6. THE FUNGAL BIOPESTICIDE METARHIZIUM ANISOPLIAE HAS AN ADJUVANT EFFECT ON THE ALLERGIC RESPONSE TO OVALBUMIN IN MICE

    Science.gov (United States)

    Background: Sensitisation to cockroaches is associated with asthma and hence, the elimination of this vermin is of interest. Metarhizium anisopliae is a parasitic fungus used as a pesticide to control cockroach infestation indoors. Previously M. anisopliae has been shown to cause...

  7. Heat-stressed Metarhizium anisopliae: Viability (in vitro) and virulence (in vivo) assessments against the tick Rhipicephalus sanguineus

    Science.gov (United States)

    The current study investigated the thermotolerance of Metarhizium anisopliae s.l. conidia from the commercial products Metarril® SP Organic and Metarril® WP. The efficacy of these M. anisopliae formulations against the tick Rhipicephalus sanguineus s.l. was studied in laboratory under optimum or hea...

  8. Differential susceptibility of adults and nymphs of Blattella germanica (L.) (Blattodea: Blattellidae) to infection by Metarhizium anisopliae and assessment of delivery strategies.

    Science.gov (United States)

    Lopes, R B; Alves, S B

    2011-01-01

    Microbial insecticides for cockroach control, such as those containing entomopathogenic fungi, may be an alternative to reduce contamination by chemicals in housing and food storage environments. Virulence of isolate ESALQ1037 belonging to the Metarhizium anisopliae complex against nymphs and adults of Blattella germanica (L.), and its infectivity following exposure of insects to a contaminated surface or to M. anisopliae-bait were determined under laboratory conditions. Estimated LD50 15 d following topical inoculation was 2.69 x 10(5) conidia per adult, whereas for nymphs the maximum mortality was lower than 50%. Baits amended with M. anisopliae conidia had no repellent effect on targets; adult mortality was inferior to 25%, and nymphs were not susceptible. All conidia found in the digestive tract of M. anisopliae-bait fed cockroaches were unviable, and bait-treated insects that succumbed to fungal infection showed a typical mycelial growth on mouthparts and front legs, but not on the hind body parts. As opposed to baits, the use of a M. anisopliae powdery formulation for surface treatment was effective in attaining high mortality rates of B. germanica. Both nymphs and adults were infected when this delivery strategy was used, and mycelia growth occurred all over the body surface. Our results suggest that the development of powders or similar formulations of M. anisopliae to control B. germanica may provide faster and better results than some of the strategies based on baits currently available.

  9. Differential susceptibility of adults and nymphs of Blattella germanica (L.) (Blattodea: Blattellidae) to infection by Metarhizium anisopliae and assessment of delivery strategies

    Energy Technology Data Exchange (ETDEWEB)

    Lopes, R.B., E-mail: rblopes@cenargen.embrapa.b [EMBRAPA Recursos Geneticos e Biotecnologia, Brasilia, DF (Brazil); Alves, S.B. [Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP), Piracicaba, SP (Brazil). Dept. de Entomologia e Acarologia

    2011-05-15

    Microbial insecticides for cockroach control, such as those containing entomopathogenic fungi, may be an alternative to reduce contamination by chemicals in housing and food storage environments. Virulence of isolate ESALQ1037 belonging to the Metarhizium anisopliae complex against nymphs and adults of Blattella germanica (L.), and its infectivity following exposure of insects to a contaminated surface or to M. anisopliae-bait were determined under laboratory conditions. Estimated LD50 15 d following topical inoculation was 2.69 x 105 conidia per adult, whereas for nymphs the maximum mortality was lower than 50%. Baits amended with M. anisopliae conidia had no repellent effect on targets; adult mortality was inferior to 25%, and nymphs were not susceptible. All conidia found in the digestive tract of M. anisopliae-bait fed cockroaches were unviable, and bait-treated insects that succumbed to fungal infection showed a typical mycelial growth on mouthparts and front legs, but not on the hind body parts. As opposed to baits, the use of a M. anisopliae powdery formulation for surface treatment was effective in attaining high mortality rates of B. germanica. Both nymphs and adults were infected when this delivery strategy was used, and mycelia growth occurred all over the body surface. Our results suggest that the development of powders or similar formulations of M. anisopliae to control B. germanica may provide faster and better results than some of the strategies based on baits currently available. (author)

  10. Cloning and functional analysis of the second geranylgeranyl diphosphate synthase gene influencing helvolic acid biosynthesis in Metarhizium anisopliae.

    Science.gov (United States)

    Singkaravanit, Suthitar; Kinoshita, Hiroshi; Ihara, Fumio; Nihira, Takuya

    2010-07-01

    A gene (ggs2) having high similarity to the geranylgeranyl diphosphate synthase (GGPP synthase) gene was cloned from Metarhizium anisopliae NAFF635007. The ggs2 gene (1,239-bp open reading frame with no intron) encoded a protein of 412 amino acids, and the transcription occurred only after late log-phase during the growth. Gene disruption of ggs2, performed to clarify the function in M. anisopliae, resulted in decreased GGPP synthase activity together with a slight delay of sporulation. An high performance liquid chromatography (HPLC) comparison of compound profiles between the wild-type strain and the disruptant revealed that a compound was abolished by the ggs2 disruption. Purification and structural elucidation by 1H-NMR and mass spectrometry analyses revealed that the lost compound is helvolic acid. Furthermore, the pathogenicity assay against two species of insect larvae revealed that the ggs2-disruptant possessed much weaker toxicity than the wild-type strain. Based on these results, it was concluded that ggs2 encodes the GGPP synthase influencing the biosynthesis of secondary metabolites in various species, including helvolic acid in M. anisopliae. To the best of our knowledge, this is the first report to identify a GGPP synthase gene related to secondary metabolism in entomopathogenic fungi.

  11. Differential pathogenicity of Metarhizium anisopliae and the control of the sugarcane root spittlebug Mahanarva fimbriolata

    Directory of Open Access Journals (Sweden)

    Patricia Vieira Tiago

    2011-06-01

    Full Text Available In order to assess the effectiveness of Metarhizium anisopliae var. anisopliae (Metschnikoff Sorokin isolates in controlling the sugarcane root spittlebug Mahanarva fimbriolata (Stal (Hemiptera: Cercopidae, nine isolates obtained from a single geographical region were studied. 'Confirmed cumulative' and 'corrected cumulative' spittlebug mortality rates were measured for each of the isolates. Based on the confirmed mortality curve, the isolates URM5946, URM5951 and URM6033 were considered to be potentially the most effective in a biological control program for M. fimbriolata.

  12. DNA polymorphism and total protein in mutants of Metarhizium anisopliae var. Anisopliae (Metsch.) Sorokin strain E9 Polimorfismo de DNA e proteína total em mutantes da linhagem E9 de Metarhizium anisopliae var. anisopliae (Metsch.) Sorokin

    OpenAIRE

    Laurineide Lopes de Carvalho Freire; Ana Bolena Lima da Costa; Larissa Brandão Góes; Neiva Tinti de Oliveira

    2001-01-01

    Five mutants (MaE10, MaE27, MaE24, MaE41 e MaE49) of Metarhizium anisopliae wild strain E9 were analysed for DNA profile through the RAPD technique and for changes in total protein content by spectrophotometry, polyacrylamide gel electrophoresis and densitometry. The pattern of RAPD markers showed genetic polymorphism among the strains: out of twenty primers seven were selected, producing 113 bands. Forty seven bands were present in all strains (41.6% of monomorphic bands) and 66 showed polym...

  13. Different Effects of Metarhizium anisopliae Strains IMI330189 and IBC200614 on Enzymes Activities and Hemocytes of Locusta migratoria L.

    Science.gov (United States)

    Cao, Guangchun; Jia, Miao; Zhao, Xia; Wang, Lei; Tu, Xiongbing; Wang, Guangjun; Nong, Xiangqun; Zhang, Zehua

    2016-01-01

    Background Metarhizium is an important class of entomopathogenic fungi in the biocontrol of insects, but its virulence is affected by insect immunity. To clarify the mechanism in virulence of Metarhizium, we compared the immunological differences in Locusta migratoria L. when exposed to two strains of Metarhizium anisopliae (Ma). Results The virulence of Ma IMI330189 was significantly higher than that of Ma IBC200614 to locust, and IMI330189 overcame the hemocytes and began destroying the hemocytes of locust at 72 h after spray, while locust is immune to IBC200614. IMI330189 could overcome the humoral immunity of locust by inhibiting the activities of phenol oxidase (PO), esterases, multi-function oxidases (MFOs) and acetylcholinesterases in locust while increasing the activities of glutathione-S-transferases (GSTs), catalase and aryl-acylamidase (AA). However IBC200614 inhibit the activities of GSTs and AA in locust and increase the activities of MFOs, PO, superoxide dismutase, peroxidase and chitinase in locust. The changes of enzymes activities in period of infection showed that the time period between the 2nd and the 5th day after spray is critical in the pathogenic process. Conclusion These results found the phenomenon that Ma initiatively broke host hemocytes, revealed the correlation between the virulence of Ma and the changes of enzymes activities in host induced by Ma, and clarified the critical period in the infection of Ma. So, these results should provide guidance for the construction of efficient biocontrol Ma strains. PMID:27227835

  14. Different Effects of Metarhizium anisopliae Strains IMI330189 and IBC200614 on Enzymes Activities and Hemocytes of Locusta migratoria L.

    Directory of Open Access Journals (Sweden)

    Guangchun Cao

    Full Text Available Metarhizium is an important class of entomopathogenic fungi in the biocontrol of insects, but its virulence is affected by insect immunity. To clarify the mechanism in virulence of Metarhizium, we compared the immunological differences in Locusta migratoria L. when exposed to two strains of Metarhizium anisopliae (Ma.The virulence of Ma IMI330189 was significantly higher than that of Ma IBC200614 to locust, and IMI330189 overcame the hemocytes and began destroying the hemocytes of locust at 72 h after spray, while locust is immune to IBC200614. IMI330189 could overcome the humoral immunity of locust by inhibiting the activities of phenol oxidase (PO, esterases, multi-function oxidases (MFOs and acetylcholinesterases in locust while increasing the activities of glutathione-S-transferases (GSTs, catalase and aryl-acylamidase (AA. However IBC200614 inhibit the activities of GSTs and AA in locust and increase the activities of MFOs, PO, superoxide dismutase, peroxidase and chitinase in locust. The changes of enzymes activities in period of infection showed that the time period between the 2nd and the 5th day after spray is critical in the pathogenic process.These results found the phenomenon that Ma initiatively broke host hemocytes, revealed the correlation between the virulence of Ma and the changes of enzymes activities in host induced by Ma, and clarified the critical period in the infection of Ma. So, these results should provide guidance for the construction of efficient biocontrol Ma strains.

  15. Adulticidal and larvicidal activity of Beauveria bassiana and Metarhizium anisopliae against housefly, Musca domestica (Diptera: Muscidae), in laboratory and simulated field bioassays.

    Science.gov (United States)

    Mishra, Sapna; Kumar, Peeyush; Malik, Anushree; Satya, Santosh

    2011-06-01

    The susceptibility of the adult and larval stage of housefly, Musca domestica L. (Diptera: Muscidae), to two entomopathogenic fungi, Metarhizium anisopliae (Metsch.) Sor. and Beauveria bassiana (Bals.) Vuill., was evaluated under laboratory and simulated field bioassays. Bioassays on adult houseflies were carried out at different conidial concentrations ranging from 10(3) to 10(9) conidia/ml in petri plate and minichamber assays. Absolute mortality was observed within 4-5 days at all the concentrations tested. M. anisopliae was found to be more effective with LC(50) of 6.75 × 10(7) conidia/ml compared with 1.21 × 10(8) conidia/ml of B. bassiana in petri plate bioassay. Similar trend was observed in minichamber bioassay. Larvicidal activity evaluated through petri plate bioassay also indicated that M. anisopliae was more effective larvicide with LC(50) of 4.1 × 10(8) conidia/ml as against 3.31 × 10(9) conidia/ml of B. bassiana. Larvicidal activity was further evaluated in simulated field condition of decaying waste matrix using dry conidial formulations (10(8) conidia/g) of both the fungi. Larval mortality obtained in this assay was 43% (B. bassiana) and 63% (M. anisopliae). Remarkably better performance of M. anisopliae as an adulticidal and larvicidal agent over B. bassiana in laboratory bioassays as well as simulated field conditions suggests that it may have good potential to become part of an integrated housefly control program.

  16. Characterization of a newly discovered China variety of Metarhizium anisopliae (M. anisopliae var. dcjhyium) for virulence to termites, isoenzyme, and phylogenic analysis.

    Science.gov (United States)

    Dong, Changjin; Zhang, Jiamin; Chen, Wuguo; Huang, Hai; Hu, Yuanyang

    2007-01-01

    The efficacy of a new virulent Metarhizium anisopliae variety (M. anisopliae var. dcjhyium, DQ288247) obtained from Odontotermes formosanus in China was evaluated against the subterranean termite, O. formosanus, in the laboratory. The new variety was compared with four other virulent M. anisopliae isolates and was found to be highly infectious and virulent against termites. M. anisopliae var. dcjhyium could cause approximately 100% mortality of termites 3 days post-inoculation in the concentration of 3x10(8) conidia/ml. There were also differences in relative hyhal growth and isoenzymes. M. anisopliae var. dcjhyium showed a different isoenzyme band pattern from the four isolates of M. anisopliae (AB027337, AB099510, AB099941 and AF280631). The phylogenetic tree of the 18S rDNA sequences revealed the taxonomic and evolutionary position of M. anisopliae var. dcjhyium. M. anisopliae var. dcjhyium and four isolates of M. anisopliae formed a monophyletic group, supported by a 99% bootstrap value. M. anisopliae var. dcjhyium formed a distinct variety, which had a special characterization of unique bands of isoenzyme, high virulence and low repellency against termites when compared with four other isolates of M. anisopliae.

  17. A Field Experiment to Assess the Rate of Infestation in Honey Bee Populations of Two Metarhizium anisopliae Isolates on Varroa destructor (Acari: Mesostigmata.

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    Khodadad Pirali-Kheirabadi

    2013-06-01

    Full Text Available The protective effect of two isolates of an entomopathogenic fungus, Metarhizium anisopliae (DEMI 002 and Iran 437C on the adult stage of Varroa destructor was evaluated in comparison with fluvalinate strips in the field.A total of 12 honey bee colonies were provided from an apiculture farm. The selected hives were divided into 4 groups (3 hives per group. The first group was the control, treated with distilled water. The other two groups were exposed to different fungi (M. anisopliae isolates DEMI 002 and Iran 437C and the last group was treated with one strip of fluvalinate per colony. The number of fallen mites was counted using sticky traps during a 6-day period, six days before and after treatments. A fungal suspension at a concentration of 5× 10(6 conidia/mL was sprayed onto the frames and the number of fallen mites was counted.Metarhizium anisopliae DEMI 002 and Iran 437C isolates were as effective (i.e., caused as much mite fall as the fluvalinate strip in controlling bee colonies than no treatment.Both M. anisopliae isolates are promising candidates as agents in the control of Varroa mites under field conditions. Isolate DEMI 002 can be considered as a possible non-chemical biocontrol agent for controlling bee infestation with V. destructor in the field. In order to substantiate this hypothesis, tests are currently being performed using larger colonies and larger doses than tested in the present study in our beekeeping.

  18. Dissemination of Metarhizium anisopliae of low and high virulence by mating behavior in Aedes aegypti

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    Rodriguez-Perez Mario A

    2011-09-01

    Full Text Available Abstract Background Dengue is a viral disease transmitted by Aedes mosquitoes. It is a threat for public health worldwide and its primary vector Aedes aegypti is becoming resistant to chemical insecticides. These factors have encouraged studies to evaluate entomopathogenic fungi against the vector. Here we evaluated mortality, infection, insemination and fecundity rates in A. aegypti females after infection by autodissemination with two Mexican strains of Metarhizium anisopliae. Methods Two M. anisopliae strains were tested: The Ma-CBG-1 least virulent (lv, and the Ma-CBG-2 highly virulent (hv strain. The lv was tested as non mosquito-passed (NMP, and mosquito-passed (MP, while the hv was examined only as MP version, therefore including the control four treatments were used. In the first bioassay virulence of fungal strains towards female mosquitoes was determined by indirect exposure for 48 hours to conidia-impregnated paper. In the second bioassay autodissemination of fungal conidia from fungus-contaminated males to females was evaluated. Daily mortality allowed computation of survival curves and calculation of the LT50 by the Kaplan-Meier model. All combinations of fungal sporulation and mating insemination across the four treatments were analyzed by χ2. The mean fecundity was analyzed by ANOVA and means contrasted with the Ryan test. Results Indirect exposure to conidia allowed a faster rate of mortality, but exposure to a fungal-contaminated male was also an effective method of infecting female mosquitoes. All females confined with the hv strain-contaminated male died in fifteen days with a LT50 of 7.57 (± 0.45 where the control was 24.82 (± 0.92. For the lv strain, it was possible to increase fungal virulence by passing the strain through mosquitoes. 85% of females exposed to hv-contaminated males became infected and of them just 10% were inseminated; control insemination was 46%. The hv strain reduced fecundity by up to 99%, and the lv

  19. Pathogenicity of a new China variety of Metarhizium anisopliae (M. Anisopliae var. Dcjhyium) to subterranean termite Odontotermes formosanus.

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    Dong, Changjin; Zhang, Jiamin; Huang, Hai; Chen, Wuguo; Hu, Yuanyang

    2009-01-01

    The pathogenicity of a new China variety of Metarhizium anisopliae (M. anisopliae var. dcjhyium) against the subterranean termite Odontotermes formosanus and the effect of the fungal fermented solution on hemolymph intracellular calcium were studied in laboratory. Conidia from the M. anisopliae var. dcjhyium were highly virulent for O. formosanus causing approximately 100% mortality 3 days post-inoculation in the concentration of 3x10(8) conidia/ml. The conidial clumps with conidial chains distributed on the cadavers of termite. When the termite was treated with 3x10(5) conidia/ml for 2 days, two constitutive proteins (91 and 105kDa) disappeared and a new specific protein (40kDa) appeared in the hemolymph of survivors relative to the controls. Hemolymph cells treated by the fungal fermented solution had a significantly higher level of intracellular calcium than controls 30min after treatment (x1.7). When the termite O. formosanus was infected by the entomopathogenic fungus M. anisopliae var. dcjhyium, hyphae invaded the integument and body cavity of the termite; well-developed muscles and fat tissue in the thorax of termite were decomposed and absorbed by hyphae, and formed the net structure; Hyphae seriously destroyed hemolymph, various tissues, pipelines and produced large number of conidia in the body of termite.

  20. Formation of exudate droplets by Metarhizium anisopliae and the presence of destruxins.

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    Hutwimmer, Stefan; Wang, Hui; Strasser, Hermann; Burgstaller, Wolfgang

    2010-01-01

    Nutritional conditions causing droplet exudation by Metarhizium anisopliae var. anisopliae were studied. Exudation in droplets occurred only on media with more than one carbon source and was highly dependent on the ratio of a well metabolized sugar such as trehalose and a nonpreferred sugar, in particular arabinose. Exuded droplets contained destruxin A, B and E in concentrations similar to those on submerged culture on Czapek Dox medium with equivalent C:N ratios but was clearly less than previously reported on standard Czapek Dox or Sabouraud dextrose broth. Destruxins also were found in agar samples from directly below mycelium and from up to 2 cm from the colony edge. Exudates retrieved from different media were proven to have Pr1 protease-related enzyme activity. Additional HPLC analysis indicated that droplets from diverse media did not differ in their sugar and acid content. A hypothesis is presented regarding the trigger for guttation in Metarhizium during growth under these conditions.

  1. DNA polymorphism and total protein in mutants of Metarhizium anisopliae var. Anisopliae (Metsch.) Sorokin strain E9

    OpenAIRE

    Freire,Laurineide Lopes de Carvalho; Costa,Ana Bolena Lima da; Góes,Larissa Brandão; Oliveira,Neiva Tinti de

    2001-01-01

    Five mutants (MaE10, MaE27, MaE24, MaE41 e MaE49) of Metarhizium anisopliae wild strain E9 were analysed for DNA profile through the RAPD technique and for changes in total protein content by spectrophotometry, polyacrylamide gel electrophoresis and densitometry. The pattern of RAPD markers showed genetic polymorphism among the strains: out of twenty primers seven were selected, producing 113 bands. Forty seven bands were present in all strains (41.6% of monomorphic bands) and 66 showed polym...

  2. Sustainable control of taro beetles via scoliid wasps\\ud and Metarhizium anisopliae

    OpenAIRE

    2014-01-01

    Taro Scarab beetles (Papuana uninodis, Coleoptera: Scarabaeidae) inflict severe damage on important root crops and plants such as Taro or Cocoyam, yam, sweet potatoes, oil palm and coffee tea plants across Africa and Asia resulting in economic hardship and starvation in some nations. Scoliid wasps and Metarhizium anisopliae fungus - bio-control agents; are shown to be able to control the population of Scarab beetle adults and larvae using a newly created simulation model based on non-linear o...

  3. Genomic analyses and transcriptional profiles of the glycoside hydrolase family 18 genes of the entomopathogenic fungus Metarhizium anisopliae.

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    Junges, Ângela; Boldo, Juliano Tomazzoni; Souza, Bárbara Kunzler; Guedes, Rafael Lucas Muniz; Sbaraini, Nicolau; Kmetzsch, Lívia; Thompson, Claudia Elizabeth; Staats, Charley Christian; de Almeida, Luis Gonzaga Paula; de Vasconcelos, Ana Tereza Ribeiro; Vainstein, Marilene Henning; Schrank, Augusto

    2014-01-01

    Fungal chitin metabolism involves diverse processes such as metabolically active cell wall maintenance, basic nutrition, and different aspects of virulence. Chitinases are enzymes belonging to the glycoside hydrolase family 18 (GH18) and 19 (GH19) and are responsible for the hydrolysis of β-1,4-linkages in chitin. This linear homopolymer of N-acetyl-β-D-glucosamine is an essential constituent of fungal cell walls and arthropod exoskeletons. Several chitinases have been directly implicated in structural, morphogenetic, autolytic and nutritional activities of fungal cells. In the entomopathogen Metarhizium anisopliae, chitinases are also involved in virulence. Filamentous fungi genomes exhibit a higher number of chitinase-coding genes than bacteria or yeasts. The survey performed in the M. anisopliae genome has successfully identified 24 genes belonging to glycoside hydrolase family 18, including three previously experimentally determined chitinase-coding genes named chit1, chi2 and chi3. These putative chitinases were classified based on domain organization and phylogenetic analysis into the previously described A, B and C chitinase subgroups, and into a new subgroup D. Moreover, three GH18 proteins could be classified as putative endo-N-acetyl-β-D-glucosaminidases, enzymes that are associated with deglycosylation and were therefore assigned to a new subgroup E. The transcriptional profile of the GH18 genes was evaluated by qPCR with RNA extracted from eight culture conditions, representing different stages of development or different nutritional states. The transcripts from the GH18 genes were detected in at least one of the different M. anisopliae developmental stages, thus validating the proposed genes. Moreover, not all members from the same chitinase subgroup presented equal patterns of transcript expression under the eight distinct conditions studied. The determination of M. anisopliae chitinases and ENGases and a more detailed study concerning the enzymes

  4. Genomic analyses and transcriptional profiles of the glycoside hydrolase family 18 genes of the entomopathogenic fungus Metarhizium anisopliae.

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    Ângela Junges

    Full Text Available Fungal chitin metabolism involves diverse processes such as metabolically active cell wall maintenance, basic nutrition, and different aspects of virulence. Chitinases are enzymes belonging to the glycoside hydrolase family 18 (GH18 and 19 (GH19 and are responsible for the hydrolysis of β-1,4-linkages in chitin. This linear homopolymer of N-acetyl-β-D-glucosamine is an essential constituent of fungal cell walls and arthropod exoskeletons. Several chitinases have been directly implicated in structural, morphogenetic, autolytic and nutritional activities of fungal cells. In the entomopathogen Metarhizium anisopliae, chitinases are also involved in virulence. Filamentous fungi genomes exhibit a higher number of chitinase-coding genes than bacteria or yeasts. The survey performed in the M. anisopliae genome has successfully identified 24 genes belonging to glycoside hydrolase family 18, including three previously experimentally determined chitinase-coding genes named chit1, chi2 and chi3. These putative chitinases were classified based on domain organization and phylogenetic analysis into the previously described A, B and C chitinase subgroups, and into a new subgroup D. Moreover, three GH18 proteins could be classified as putative endo-N-acetyl-β-D-glucosaminidases, enzymes that are associated with deglycosylation and were therefore assigned to a new subgroup E. The transcriptional profile of the GH18 genes was evaluated by qPCR with RNA extracted from eight culture conditions, representing different stages of development or different nutritional states. The transcripts from the GH18 genes were detected in at least one of the different M. anisopliae developmental stages, thus validating the proposed genes. Moreover, not all members from the same chitinase subgroup presented equal patterns of transcript expression under the eight distinct conditions studied. The determination of M. anisopliae chitinases and ENGases and a more detailed study

  5. Secondary metabolite gene clusters in the entomopathogen fungus Metarhizium anisopliae: genome identification and patterns of expression in a cuticle infection model

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    Nicolau Sbaraini

    2016-10-01

    Full Text Available Abstract Background The described species from the Metarhizium genus are cosmopolitan fungi that infect arthropod hosts. Interestingly, while some species infect a wide range of hosts (host-generalists, other species infect only a few arthropods (host-specialists. This singular evolutionary trait permits unique comparisons to determine how pathogens and virulence determinants emerge. Among the several virulence determinants that have been described, secondary metabolites (SMs are suggested to play essential roles during fungal infection. Despite progress in the study of pathogen-host relationships, the majority of genes related to SM production in Metarhizium spp. are uncharacterized, and little is known about their genomic organization, expression and regulation. To better understand how infection conditions may affect SM production in Metarhizium anisopliae, we have performed a deep survey and description of SM biosynthetic gene clusters (BGCs in M. anisopliae, analyzed RNA-seq data from fungi grown on cattle-tick cuticles, evaluated the differential expression of BGCs, and assessed conservation among the Metarhizium genus. Furthermore, our analysis extended to the construction of a phylogeny for the following three BGCs: a tropolone/citrinin-related compound (MaPKS1, a pseurotin-related compound (MaNRPS-PKS2, and a putative helvolic acid (MaTERP1. Results Among 73 BGCs identified in M. anisopliae, 20 % were up-regulated during initial tick cuticle infection and presumably possess virulence-related roles. These up-regulated BGCs include known clusters, such as destruxin, NG39x and ferricrocin, together with putative helvolic acid and, pseurotin and tropolone/citrinin-related compound clusters as well as uncharacterized clusters. Furthermore, several previously characterized and putative BGCs were silent or down-regulated in initial infection conditions, indicating minor participation over the course of infection. Interestingly, several up

  6. Total lipids and fatty acids of strains of Metarhizium anisopliae

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    Pupin Antonio Marcos

    2000-01-01

    Full Text Available Two growth stages, conidia (C and mycelium (M, and two media, minimal medium (MM and complete medium (MC, were compared in 10 strains of M. anisopliae, and two strains of M. anisopliae var. majus were similar in percentages of total lipids. Tukey test for average of lipid content in conidia (C and mycelia (M cultured on minimal (MM and complete (MC media showed significant differences between means at the 5% level for mycelia and conidia, indicating variability in total lipid production and storage during growth. Strains 5 and 7, both variety majus, did not present sizable differences from variety anisopliae. For fatty acids, C18:1 and C18:2, oleic and linoleic, respectively, the differences were all highly significant (p= 1% with the highest means being obtained for conidia for fatty acid C18:1 and for myclelia for fatty acid C18:2.

  7. Metarhizium anisopliae lipolytic activity plays a pivotal role in Rhipicephalus (Boophilus) microplus infection.

    Science.gov (United States)

    Beys da Silva, Walter O; Santi, Lucélia; Schrank, Augusto; Vainstein, Marilene H

    2010-01-01

    Lipases secreted by Metarhizium anisopliae, an important biological control agent, could potentially be involved in the host infection process. Here, we present the activity profile during the host infection process and the effect of lipase activity inhibitor ebelactone B on infection. The previous treatment of spores with lipase activity inhibitor, ebelactone B, completely inhibited lipolytic activity and prevented the infection of the Rhipicephalus (Boophilus) microplus host. The results herein presented prove, for the first time, the importance of lipase activity in M. anisopliae host infection process. The filamentous fungus Metarhizium anisopliae is one of the most important and studied biological agents for the control of several arthropod pests, including the cattle tick Rhipicephalus (Boophilus) microplus. Lipases secreted by M. anisopliae could potentially be involved in the host infection process. This work presents the activity profile during the host infection process and the effect of lipase activity inhibitor ebelactone B on infection. During the course of tick exposure to spores (6-120 h) lipase activity increased from 0.03 ± 0.00 U to 0.312 ± 0.068 U using rho NP palmitate as substrate. In zymograms, bands of lipase activity were detected in ticks treated with spores without inhibitor. The previous treatment of spores with lipase activity inhibitor, ebelactone B, completely inhibited lipolytic activity, at all times specified, and prevented the infection of the R. microplus host. Spores treated with the inhibitor did not germinate on the tick, although this effect was not observed in the culture medium. The results herein presented prove, for the first time, the importance of lipase activity in M. anisopliae host infection process.

  8. Susceptibilidade de larvas de Cerotoma arcuata Olivier (Coleoptera: Chrysomelidae a Beauveria bassiana (Bals. Vuillemin, Metarhizium anisopliae (Metsch. Sorokin e Bacillus thuringiensis Berliner Susceptibility of Cerotoma arcuata Olivier (Coleoptera: Chrysomelidae larvae to Beauveria bassiana (Bals. Vuillemin, Metarhizium anisopliae (Metsch. Sorokin and Bacillus thuringiensis Berliner

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    Maria Lucia França Teixeira

    2007-02-01

    Full Text Available Larvas de 2° instar de Cerotoma arcuata foram avaliadas em relação à susceptibilidade aos fungos entomopatogênicos Beauveria bassiana, Metarhizium anisopliae e a bactéria Bacillus thuringiensis com as toxinas Cry3. Os insetos adultos foram mantidos em gaiolas e alimentados com plântulas de feijão (Phaseolus vulgaris L. e as larvas em "gerbox" com cotilédones de plântulas de feijão recém-germinadas. Das oito estirpes de B. bassiana avaliadas, CG 156 e CG 213 causaram 100% de mortalidade das larvas, as duas estirpes de M. anisopliae CG 210 e CG 321 foram patogênicas, eliminando 80 e 100% das larvas de C. arcuata, e, das cinco estirpes de B. thuringiensis testadas, o isolado CG 940 causou 70% de mortalidade das larvas.Second instar larvae of Cerotoma arcuata were evaluated concerning the susceptibility to fungi Beauveria bassiana and Metarhizium anisopliae and Bacillus thuringiensis strains containing Cry3 toxin. Adults of C. arcuata were kept in large cages and fed on bean seedlings and the larvae were reared in ‘gearbox’ feeding on germinated Phaseolus bean cotyledons. Strains CG 156 and CG 213 of B. bassiana killed 100% of the insect larvae and strains CG 210 and CG 321 of M. anisopliae killed 80 and 100% of the insect larvae. Strain CG 940 of B. thuringiensis killed 70% of the insect larvae.

  9. Biocontrol Potential of Metarhizium anisopliae (Metsch. Sorokin (Deuteromycotina: Hyphomycetes against Ailanthus Defoliator, Eligma narcissus (Cram.

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    N.S. Bai

    2015-09-01

    Full Text Available Eligma narcissus is recognised as a serious pest of Ailanthus in Southern India and defoliation of Ailanthus by this pest causes apparent loss of growth increment. The common control methods for this pest is mostly insecticides and the concern about the environmental effects of chemical insecticides, has emphasised the use of environmentally more benign microbial agents. Among entomopathogens, Fungi are the most explored and often act as important natural control agents that limit insect populations. On this point of view, Bio efficacy of 25 isolates of Metarhizium anisopliae was assessed to establish their virulence against E. narcissus in the laboratory and effective formulations of two potent isolates were subsequently evaluated in the field. MIS7 and MIS13 were more effective among the different isolates evaluated against E. narcissus. The median lethal concentration (LC50 of all the isolates ranged from 6.46×105 conidia/ml to 628.92×105 conidia/ml. Median lethal concentration of (LT50 of 4.9 and 5.4 days were recorded for MIS7 and MIS13 respectively at a concentration of 1× 107 conidia/ml. Virulence tests of the isolates MIS7 and MIS13 and 0.5% Pongamia pinnata seed oil, individually and in different combinations, indicated improved efficacy of the isolates when used in combination and also when combined with seed oil. Formulations composed of “MIS7+MIS13+0.5% Pongamia pinnata seed oil” and “MIS7+MIS13” proved to be superior against E. narcissus, causing 76.30% and 93.93% mortality, respectively. Field evaluation of the formulation MIS7+MIS13+0.5% Pongamia pinnata seed oil recorded 5.79 larvae per plant resulting in 60.53% reduction of infestation while the formulation, MIS7+MIS13 showed 53.76% reduction of infestation with 6.56 larvae per plant. The observations from this study suggest the prospects of using the entomopathogenic fungus, M. anisopliae for the control of E. narcissus.

  10. Selection of Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato isolates as microbial control agents against the boll weevil (Anthonomus grandis) in Argentina.

    Science.gov (United States)

    Nussenbaum, A L; Lecuona, R E

    2012-05-01

    The boll weevil (Anthonomus grandis) is the main pest of cotton in the Americas. The aim of this work was to evaluate isolates of the entomopathogenic fungi Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato virulent against A. grandis. Screening was performed to evaluate the pathogenicity of 28 isolates of M. anisopliae s.l. and 66 isolates of B. bassiana s.l. against boll weevil adults. To select the isolates, LC(50) values of the most virulent isolates were calculated, and compatibility between the fungi and insecticides was studied. In addition, the effects of these isolates on the feeding behavior of the adults were evaluated. Isolates Ma 50 and Ma 20 were the most virulent against A. grandis and their LC(50) values were 1.13×10(7) and 1.20×10(7) conidia/ml, respectively. In addition, these isolates were compatible with pyrethroid insecticides, but none with endosulfan. On the other hand, infected females reduced the damage caused by feeding on the cotton squares and their weight gain. This shows that entomopathogenic fungi cause mortality in the insects, but also these fungi could influence the feeding behavior of the females. In summary, these results indicate the possibility of the use of M. anisopliae s.l. as a microbiological control agent against boll weevils. Also, this species could be included in an Integrated Pest Management program.

  11. Metarhizium anisopliae and Trichoderma viride control colonies of Atta cephalotes in field better than a chemical insecticide

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    Elkin López Arismendy

    2011-12-01

    Full Text Available The leaf-cutting ant Atta cephalotes is an economically important pest in agriculture. These ants use the material they cut to cultivate a fungus from which they fed. In this study, baits prepared with the entomopathogenic fungus Metarhizium anisopliae strain M-137, Trichoderma viride strain T-26, antagonist of the symbiont fungus of A. cephaloies, and a combination of both fungi were applied in order to control A. cephalotes colonies under field conditions. In addition, the insecticide activity of the baits was compared to a chemical product, Pirimifos Metil, which was applied with an air pump. The ants did not detect the fungal agents contained in the baits, and introduced then into their nests without awakening defensive behaviors. The mortality of the bait-treated nests was higher than 80% while the treatment with Pirimifos Metil was effective in only 60% of the nests. Additionally, a week after the application of these treatments, changes in the insects' behavior were observed, reflected mainly in the absence of foraging activity. In conclusion, M. anisopliae was effective in controlling A. cephalotes colonies, and superior to the chemical product Pirimifos Metil. 

  12. Comparative transcriptomic analysis of the heat stress response in the filamentous fungus Metarhizium anisopliae using RNA-Seq.

    Science.gov (United States)

    Wang, Zhang-Xun; Zhou, Xia-Zhi; Meng, Hui-Min; Liu, Yu-Jun; Zhou, Quan; Huang, Bo

    2014-06-01

    The entomopathogenic fungus Metarhizium anisopliae is widely used for biological control of a variety of insect pests. The effectiveness of the microbial pest control agent, however, is limited by poor thermotolerance. The molecular mechanism underlying the response to heat stress in the conidia of entomopathogenic fungi remains unclear. Here, we conducted high-throughput RNA-Seq to analyze the differential gene expression between control and heat treated conidia of M. anisopliae at the transcriptome level. RNA-Seq analysis generated 6,284,262 and 5,826,934 clean reads in the control and heat treated groups, respectively. A total of 2,722 up-regulated and 788 down-regulated genes, with a cutoff of twofold change, were identified by expression analysis. Among these differentially expressed genes, many were related to metabolic processes, biological regulation, cellular processes and response to stimuli. The majority of genes involved in endocytic pathways, proteosome pathways and regulation of autophagy were up-regulated, while most genes involved in the ribosome pathway were down-regulated. These results suggest that these differentially expressed genes may be involved in the heat stress response in conidia. As expected, significant changes in expression levels of genes encoding heat shock proteins and proteins involved in trehalose accumulation were observed in conditions of heat stress. These results expand our understanding of the molecular mechanisms of the heat stress response of conidia and provide a foundation for future investigations.

  13. Metarhizium anisopliae chitinase CHIT30 is involved in heat-shock stress and contributes to virulence against Dysdercus peruvianus.

    Science.gov (United States)

    Staats, Charley Christian; Kmetzsch, Livia; Lubeck, Irina; Junges, Angela; Vainstein, Marilene Henning; Schrank, Augusto

    2013-02-01

    Entomopathogenic fungi are able to produce several chitinases, which serve a variety of biological functions, such as fungal cell wall organization and the degradation of exogenous chitin for nutrition or insect infection processes. In this study, we analyzed the contribution of the CHIT30 chitinase from Metarhizium anisopliae in morphogenetic development and virulence as a model of chitinase function. The analysis of chi3 gene expression revealed transcript accumulation in response to heat-shock stress conditions as well as cultivation in medium supplemented with chitin. Null chi3 mutants were constructed to determine the biological role of CHIT30. No substantial differences in the secreted chitinase activity could be detected between the wild type and the Δchi3 mutant. However, both endochitinase and exochitinase activities were diminished in the mutant strain following heat-shock treatment, suggesting that CHIT30 is involved in heat-shock adaptation. Mutants lacking CHIT30 chitinase showed reduced virulence against the cotton stainer bug Dysdercus peruvianus, indicating that the CHIT30 chitinase plays a role in the infection process of M. anisopliae.

  14. Identification of genes differentially expressed by Metarhizium anisopliae growing on Locusta migratoria wings using suppression subtractive hybridization.

    Science.gov (United States)

    Zhang, Chuanbo; Xia, Yuxian; Li, Zhongyuan

    2011-05-01

    Insect-pathogenic fungi penetrate their hosts directly through the cuticle. To better understand this process, we identified genes that were up-regulated by Metarhizium anisopliae germinating and differentiating on Locusta migratoria wings using suppression subtractive hybridization (SSH). A total of 78 unique expressed sequence tags (ESTs) up-regulated more than twofold during fungal growth on locust wings were identified. Among these 78 ESTs, 30 (38.5%) shared significant similarity with NCBI annotated hypothetical proteins, 16 (20.5%) shared low similarity to known or predicted genes, might represent novel genes, and 32 (41.0%) shared significant similarity with known proteins that are involved in various cell and molecular processes such as cell metabolism, protein metabolism, stress response and defense, and cell structure and function. Semi-quantitative RT-PCR analysis of six randomly selected genes confirmed the SSH results, verifying the fidelity of the SSH data. The results of this study provide novel information on genes expressed during early stages of infection with M. anisopliae, and improve current understanding of fungal pathogenesis.

  15. Field Efficacy of a Metarhizium anisopliae-Based Attractant-Contaminant Device to Control Ceratitis capitata (Diptera: Tephritidae).

    Science.gov (United States)

    Navarro-Llopis, V; Ayala, I; Sanchis, J; Primo, J; Moya, P

    2015-08-01

    Biological control of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) using entomopathogenic fungi is being studied as a viable control strategy. The efficacy of a Metarhizium anisopliae (Metschnikoff) Sorokin (Hypocreales: Clavicipitaceae)-based attractant-contaminant device (ACD) to control C. capitata was evaluated in a medium-scale (40 ha) 2-yr field trial using a density of 24 ACD per ha. Results showed that this density was adequate to efficiently reduce fruitfly populations and that the inoculation dishes (IDs) needed replacing mid-season to provide protection for the entire season. In this study, fungal treatment was even more effective than conventional chemical treatment. Population dynamics in fungus-treated fields along with the infectivity study of field-aged IDs in the laboratory found that the ACD remained effective for at least 3 mo. The results suggest M. anisopliae-based ACD can be used to control C. capitata in the field. The implications of its use, especially as a tool in an integrated pest management program, are discussed. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  16. The influence of conidial Pr1 protease on pathogenicity potential of Metarhizium anisopliae senso latu to ticks.

    Science.gov (United States)

    Golo, Patrícia S; Santos, Huarrisson A; Perinotto, Wendell M S; Quinelato, Simone; Angelo, Isabele C; Camargo, Mariana G; Sá, Fillipe A; Massard, Carlos L; Fernandes, Éverton K K; Roberts, Donald W; Bittencourt, Vânia R E P

    2015-06-01

    Pr1 is a subtilisin-like protease produced by Metarhizium spp. entomopathogenic fungi, and it is recognized as heavily involved in the initial steps of the fungal invasion of arthropod-host cuticles. In the current study, correlation was sought between mortality of tick larvae and conidial Pr1 levels of one Metarhizium anisopliae senso latu (s.l.) isolate (CG 148). Conidia with different levels of pr1 gene expression and enzymatic activity were obtained by producing them on either artificial medium (to yield low Pr1 activity) or on Rhipicephalus microplus cadavers (to yield high Pr1 activity). Conidial proteolytic activity was assessed using N-suc-ala-ala-pro-phe-ρNA as the chromogenic substrate, and pr1 expression was profiled by qPCR using three genes (gpd, try, and tef) as reference genes. Pr1 enzymatic (proteolytic) activity on conidia obtained from tick cadavers was 36 U mg(-1) in comparison to 4 U mg(-1) on conidia from PDA medium. Also, pr1 gene expression level was ten times higher in conidia from tick cadavers compared to PDA medium. Bioassays of M. anisopliae s.l. CG 148 spores with elevated Pr1 proteolytic activity and gene expression levels did not demonstrate increased virulence (= significant change percent mortality of tick larvae). The minimal levels of Pr1 on conidia produced on artificial medium was adequate to afford high levels of virulence, and the elevated amounts of the enzyme on tick-cadaver-produced conidia did not induce elevated larval mortality. As long as some Pr1 activity was present, fungal virulence of isolate CG 148 against tick larvae was not elevated by increased levels of conidial Pr1.

  17. Pengujian Toksisitas Akut Oral Dan Dermal pada Biolarvasida Metarhizium anisopliae terhadap Tikus Putih Spraque Dawley

    Directory of Open Access Journals (Sweden)

    Deni Zulfiana

    2016-03-01

    Full Text Available Acute oral and dermal toxicity test against white rats was conducted to determine the toxicity and side effects of bio-larvacide (Metarhizium anisopliae crude extract on humans. In the oral test used a maximum dose 5000 mg/kg and dermal testing used a maximum dose of 2000 mg/kg. Dose treatment and control tested to 5 Spraque Dawley male rats. The results showed that oral treatment with a dose of 5000 mg/kg did not cause mortality and did not cause changes in anatomic pathology of viceral organs. In the dermal treatment with a dose of 2000 mg/kg did not cause mortality and did not cause changes in anatomic pathology of viceral organs. Based on these results LD50 acute oral M. anisopliae biolarvacide above 5000 mg/kg and the acute dermal is above 2000 mg/kg. It was therefore concluded that the formulation of Metarhizium anisopliae biolarvasida classified as not hazardous when used in accordance with the recommendation of the class I (WHO, 2003.

  18. Impact of moisture on survival of Aedes aegypti eggs and ovicidal activity of Metarhizium anisopliae under laboratory conditions

    OpenAIRE

    C Luz; MHH Tai; AH Santos; HHG Silva

    2008-01-01

    The effect of relative humidity (43%, 75%, 86% and > 98%) on Aedes aegypti eggs treated with Metarhizium anisopliae or water only was tested for up to a six months exposure at 25ºC. Survival of larvae inside eggs was clearly affected by the lowest humidity (43%) tested, and eclosion diminished at all humidities after increasing periods of exposure. M. anisopliae showed to have a strong ovicidal activity only at humidity close to saturation. No difference of activity was found between conidia ...

  19. Susceptibility of adult female Aedes aegypti (Diptera: Culicidae to the entomopathogenic fungus Metarhizium anisopliae is modified following blood feeding

    Directory of Open Access Journals (Sweden)

    Samuels Richard I

    2011-05-01

    Full Text Available Abstract Background The mosquito Aedes aegypti, vector of dengue fever, is a target for control by entomopathogenic fungi. Recent studies by our group have shown the susceptibility of adult A. aegypti to fungal infection by Metarhizium anisopliae. This fungus is currently being tested under field conditions. However, it is unknown whether blood-fed A. aegypti females are equally susceptible to infection by entomopathogenic fungi as sucrose fed females. Insect populations will be composed of females in a range of nutritional states. The fungus should be equally efficient at reducing survival of insects that rest on fungus impregnated surfaces following a blood meal as those coming into contact with fungi before host feeding. This could be an important factor when considering the behavior of A. aegypti females that can blood feed on multiple hosts over a short time period. Methods Female A. aegypti of the Rockefeller strain and a wild strain were infected with two isolates of the entomopathogenic fungus M. anisopliae (LPP 133 and ESALQ 818 using an indirect contact bioassay at different times following blood feeding. Survival rates were monitored on a daily basis and one-way analysis of variance combined with Duncan's post-hoc test or Log-rank survival curve analysis were used for statistical comparisons of susceptibility to infection. Results Blood feeding rapidly reduced susceptibility to infection, determined by the difference in survival rates and survival curves, when females were exposed to either of the two M. anisopliae isolates. Following a time lag which probably coincided with digestion of the blood meal (96-120 h post-feeding, host susceptibility to infection returned to pre-blood fed (sucrose fed levels. Conclusions Reduced susceptibility of A. aegypti to fungi following a blood meal is of concern. Furthermore, engorged females seeking out intra-domicile resting places post-blood feeding, would be predicted to rest for prolonged

  20. Susceptibility of adult female Aedes aegypti (Diptera: Culicidae) to the entomopathogenic fungus Metarhizium anisopliae is modified following blood feeding.

    Science.gov (United States)

    Paula, Adriano R; Carolino, Aline T; Silva, Carlos P; Samuels, Richard I

    2011-05-26

    The mosquito Aedes aegypti, vector of dengue fever, is a target for control by entomopathogenic fungi. Recent studies by our group have shown the susceptibility of adult A. aegypti to fungal infection by Metarhizium anisopliae. This fungus is currently being tested under field conditions. However, it is unknown whether blood-fed A. aegypti females are equally susceptible to infection by entomopathogenic fungi as sucrose fed females. Insect populations will be composed of females in a range of nutritional states. The fungus should be equally efficient at reducing survival of insects that rest on fungus impregnated surfaces following a blood meal as those coming into contact with fungi before host feeding. This could be an important factor when considering the behavior of A. aegypti females that can blood feed on multiple hosts over a short time period. Female A. aegypti of the Rockefeller strain and a wild strain were infected with two isolates of the entomopathogenic fungus M. anisopliae (LPP 133 and ESALQ 818) using an indirect contact bioassay at different times following blood feeding. Survival rates were monitored on a daily basis and one-way analysis of variance combined with Duncan's post-hoc test or Log-rank survival curve analysis were used for statistical comparisons of susceptibility to infection. Blood feeding rapidly reduced susceptibility to infection, determined by the difference in survival rates and survival curves, when females were exposed to either of the two M. anisopliae isolates. Following a time lag which probably coincided with digestion of the blood meal (96-120 h post-feeding), host susceptibility to infection returned to pre-blood fed (sucrose fed) levels. Reduced susceptibility of A. aegypti to fungi following a blood meal is of concern. Furthermore, engorged females seeking out intra-domicile resting places post-blood feeding, would be predicted to rest for prolonged periods on fungus impregnated black cloths, thus optimizing infection

  1. Infection of two non-target grasshoppers by the biological control agent Metarhizium anisopliae var. acridum in the Sahel

    DEFF Research Database (Denmark)

    Fisker, E. N.; Eilenberg, J.; Langewald, J.;

    2006-01-01

    Fungal isolates from grasshoppers of the family Acrididae are suspected to be less virulent to grasshoppers of the family Pyrgomorphidae. The biological control agent Metarhizium anisopliae var. acridum was isolated from an acridid and is thus hypothesized to be less virulent to pyrgomorphids....... The susceptibility of two non-target pyrgomorphids, Pyrgomorpha cognata and Poekilocerus bufonius hieroglyphicus, to M. anisopliae was tested in the field. Results show that P. cognata under field conditions is as susceptible to infection by M. anisopliae as acridids, whereas P. b. hieroglyphicus is less susceptible...

  2. Metarhizium anisopliae pathogenesis of mosquito larvae: a verdict of accidental death.

    Science.gov (United States)

    Butt, Tariq M; Greenfield, Bethany P J; Greig, Carolyn; Maffeis, Thierry G G; Taylor, James W D; Piasecka, Justyna; Dudley, Ed; Abdulla, Ahmed; Dubovskiy, Ivan M; Garrido-Jurado, Inmaculada; Quesada-Moraga, Enrique; Penny, Mark W; Eastwood, Daniel C

    2013-01-01

    Metarhizium anisopliae, a fungal pathogen of terrestrial arthropods, kills the aquatic larvae of Aedes aegypti, the vector of dengue and yellow fever. The fungus kills without adhering to the host cuticle. Ingested conidia also fail to germinate and are expelled in fecal pellets. This study investigates the mechanism by which this fungus adapted to terrestrial hosts kills aquatic mosquito larvae. Genes associated with the M. anisopliae early pathogenic response (proteinases Pr1 and Pr2, and adhesins, Mad1 and Mad2) are upregulated in the presence of larvae, but the established infection process observed in terrestrial hosts does not progress and insecticidal destruxins were not detected. Protease inhibitors reduce larval mortality indicating the importance of proteases in the host interaction. The Ae. aegypti immune response to M. anisopliae appears limited, whilst the oxidative stress response gene encoding for thiol peroxidase is upregulated. Cecropin and Hsp70 genes are downregulated as larval death occurs, and insect mortality appears to be linked to autolysis through caspase activity regulated by Hsp70 and inhibited, in infected larvae, by protease inhibitors. Evidence is presented that a traditional host-pathogen response does not occur as the species have not evolved to interact. M. anisopliae retains pre-formed pathogenic determinants which mediate host mortality, but unlike true aquatic fungal pathogens, does not recognise and colonise the larval host.

  3. Unveiling chemical defense in the rice stalk stink bug against the entomopathogenic fungus Metarhizium anisopliae.

    Science.gov (United States)

    da Silva, Rodrigo Alves; Quintela, Eliane Dias; Mascarin, Gabriel Moura; Pedrini, Nicolás; Lião, Luciano Moraes; Ferri, Pedro Henrique

    2015-05-01

    Eggs, nymphs (1st-5th instar) and adults of Tibraca limbativentris were challenged by conidial suspensions of its major fungal pathogen Metarhizium anisopliae in order to assess their susceptibility. The role of chemical defensive compounds from exocrine secretions produced by both nymphs and adults were examined for their participation on M. anisopliae infection. Although insect susceptibility to M. anisopliae followed a dose-dependent manner, adults followed by older nymphs displayed the highest resistance. Eggs were highly susceptible showing >96% fungal infection. Crude extracts isolated from metathoracic scent gland and dorsal abdominal glands of adults and nymphs, respectively, showed fungistatic effects by impairing spore germination, vegetative growth and sporulation. Gas chromatography-mass spectrometry analysis of these extracts revealed that the major components were short-chain hydrocarbons (C10-13) and unsaturated aldehydes. In vitro tests with the corresponding synthetic standards indicated compounds with greater antifungal activity including (E)-2-hexenal, (E)-2-octenal, and (E)-2-decenal, with the latter being the most deleterious to fungal fitness. We demonstrated that differential susceptibility of the rice stalk stink bug to M. anisopliae infection is age-specific and partly mediated by fungistatic properties of aldehydes, which are produced by scent glands of both nymphs and adults.

  4. In vitro interaction of Metarhizium anisopliae Ma9236 and Beauveria bassiana Bb9205 with Heterorhabditis bacteriophora HNI0100 for the control of Plutella xylostella.

    Science.gov (United States)

    Correa-Cuadros, J P; Sáenz-Aponte, A; Rodríguez-Bocanegra, M X

    2016-01-01

    The diamondback moth (Plutella xylostella) is a major pest of broccoli crops in Colombia. To control P. xylostella, we evaluated the interaction of Beauveria bassiana Bb9205 and Metarhizium anisopliae Ma9236 with Heterorhabditis bacteriophora HNI0100 and its bacterial symbiont Photorhabdus luminescens HNI0100. We used antagonism and disk diffusion assays with fungal extracts to test the interaction between symbiotic bacterium and fungi. P. luminescens inhibited the growth of B. bassiana and M. anisopliae up to 40% by the secretion of secondary metabolites, whereas fungal extracts did not inhibit P. luminescens; this explains the in vivo interactions of these biological control agents. To test the interaction between fungi and nematodes, we first inoculated the fungi followed by the nematodes on different days (0, 2, 4, and 6). We identified the type of interaction using the formula by Nishimatsu and Jackson (J Econ Entomol 91:410-418, 1998) and established that on days 0, 2 and 4 there was an antagonistic interaction, while a synergistic interaction occurred on day 6. Therefore, the use of the interaction between H. bacteriophora HNI0100 with M. anisopliae Ma9236 and B. bassiana Bb9205 is an innovative alternative for the control of P. xylostella.

  5. 球孢白僵菌和金龟子绿僵菌不同菌株对黑足角胸叶甲成虫的致病力评价%Pathogenicity evaluation of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae against adults of Basilepta melanopus ( Coleoptera: Eumolpidae)

    Institute of Scientific and Technical Information of China (English)

    何学友; 蔡守平; 童应华; 熊瑜; 黄勇; 谢家冬; 陈顺立

    2011-01-01

    In order to screen entomopathogenic fungi strains with high virulence against the new pest Basilepta melanopus on Camellia oleifera, the growth rate and sporulation of different Beauveria bassiana and Metarhizium anisopliae strains were investigated, and bioassay evaluation of B. Bassiana and M. Anisopliae against B. Melanopus adults was carried out through dipping in spore suspension. The results showed that the growth rate and sporulation of different fungi strains were significantly different. The four strains, I. E., MaYTTR-04, BbFZ-17, MaZPTR-01 and BbTK-01, had higher growth rate and sporulation than other strains. The cumulative mortality of B. Melanopus adults was gradually increased with time after inoculation with B. Bassiana and M. Anisopliae. The cumulative mortality of B. Melanopus adults reached 100% at 7 d after inoculation with B. Bassiana, while the mortality of adults at 14 d after inoculation with MaZPTR-01 and MaYTTR-04 were 80. 3% and 78. 8% , respectively. The cadaver rate of adults inoculated with B. Bassiana was significantly higher than that inoculated with M. Anisopliae. The cadaver rates of BbTK-01 and BbFZ-17 were highest, being 85.7% and 75. 8% , respectively. The median lethal time (LT50) values of B. Bassianu strains BbXJ-01, BbFZ-17 and BbTK-01 were shortest, being 3.0, 3. 3 and 3. 4 d, respectively. However, the LTXvalues of the two M. Anisopliae strains, MaZPTR-01 and MaYTTR-04, were 6.0 d and 6.2 d, respectively. The results of bioassay showed that B. Bassiana strains had higher virulence to B. Melanopus adults than M. Anisopliae, especially the two strains, BbTK-01 and BbFZ-17, had higher lethality and infection rate than other strains, and at the same time, they had the characteristics of rapid growth and high sporulation, suggesting that the two strains have great potential in biocontrol of B. Melanopus adults.%本研究在测定不同球孢白僵菌Beauveria bassiana和金龟子绿僵菌Metarhizium anisopliae菌株的生长

  6. Laboratory mortality and mycosis of adult Curculio caryae (Coleoptera: Curculionidae) following application of Metarhizium anisopliae in the laboratory and field

    Science.gov (United States)

    The pecan weevil, Curculio caryae, is a key pest of pecans. Our objective was to determine the potential of Metarhizium anisopliae to control emerging C. caryae adults. First, a laboratory test was conducted to compare four Beauveria bassiana strains (Bb GA2, BbLA3, BbMS1, and GHA) and three M. an...

  7. Cover crop and conidia delivery system impacts on soil persistence of Metarhizium anisopliae (Hypocreales:Clavicipitaceae) in sugarbeet

    Science.gov (United States)

    The sugarbeet root maggot, Tetanops myopaeformis (Röder), is a major North American pest of sugarbeet, Beta vulgaris L. Previous research suggests moderate field efficacy of the fungal entomopathogen Metarhizium anisopliae (Metch.) Sorok. against T. myopaeformis larvae. We conducted three-years of f...

  8. Bioreactor and substrate selection for solid-state cultivation of the malaria mosquito control agent Metarhizium anisopliae

    NARCIS (Netherlands)

    Breukelen, van F.R.; Haemers, S.; Wijffels, R.H.; Rinzema, A.

    2011-01-01

    Metarhizium anisopliae spores are a promising alternative to chemical insecticides against malaria mosquitoes. In-house application resulting in infection of mosquitoes with the fungus can strongly contribute to reducing malaria transmission. For such application, fungal spores need to be produced i

  9. Autodissemination of the entomopathogenic fungus Metarhizium anisopliae amongst adults of the malaria vector anopheles gambiae s.s.

    NARCIS (Netherlands)

    Scholte, E.J.; Knols, B.G.J.; Takken, W.

    2004-01-01

    Background - The entomopathogenic fungus Metarhizium anisopliae is being considered as a biocontrol agent for adult African malaria vectors. In the laboratory, work was carried out to assess whether horizontal transmission of the pathogen can take place during copulation, as this would enhance the i

  10. Exposure of bed bugs to metarhizium anisopliae, and the effect of defensive secretions on fungal growth in vitro

    Science.gov (United States)

    Bed bugs Cimex lectularius were treated with conidia of the entomopathogenic fungus Metarhizium anisopliae by topical, spray, and contact exposure. One week post-exposure, inconsistent mortalities were observed, averaging 30% across all treatment groups and replicates. Microscopic examination of top...

  11. Infection of the malaria mosquito Anopheles gambiae with the entomopathogenic fungus Metarhizium anisopliae reduces blood feeding and fecundity

    NARCIS (Netherlands)

    Scholte, E.J.; Knols, B.G.J.; Takken, W.

    2006-01-01

    The entomopathogenic fungus Metarhizium anisopliae is being considered as a biocontrol agent against adult African malaria vectors. In addition to causing significant mortality, this pathogen is known to cause reductions in feeding and fecundity in a range of insects. In the present study we investi

  12. First report of Metarhizium anisopliae IP 46 pathogenicity in adult Anopheles gambiae s.s. and An. arabiensis (Diptera; Culicidae)

    NARCIS (Netherlands)

    Mnyone, L.L.; Russell, T.L.; Lyimo, I.N.; Lwetoijera, D.W.; Kirby, M.J.; Luz, C.

    2009-01-01

    The entomopathogenic fungus Metarhizium anisopliae isolate IP 46, originating from a soil sample collected in 2001 in the Cerrado of Central Brazil, was tested for its ability to reduce the survival of adult male and female Anopheles gambiae s.s. and An. arabiensis mosquitoes. A 6-h exposure to the

  13. Controle de larvas de Boophilus microplus por Metarhizium anisopliae em pastagens infestadas artificialmente Control of Boophilus microplus larvae by Metarhizium anisopliae in artificially infested pastures

    Directory of Open Access Journals (Sweden)

    Lúcia Mara de Souza Basso

    2005-06-01

    Full Text Available O objetivo deste trabalho foi avaliar a eficiência do controle exercido por Metarhizium anisopliae na população de Boophilus microplus, em pastagens de Brachiaria brizantha, e do híbrido Tifton 85 (Cynodon spp., artificialmente infestadas com fêmeas ingurgitadas do carrapato. Trinta canteiros com 1 m² de área cada foram distribuídos aleatoriamente. Quinze foram pulverizados com esporos do fungo e quinze controles em cada forrageira, constituindo cinco repetições de cada tratamento, foram infestados com número e peso padronizados de fêmeas ingurgitadas do ácaro. Aplicou-se o fungo, na concentração de 1,8x10(8 conídios mL-1, em três situações: pulverização antes da infestação com o carrapato, após a infestação e posterioriormente à emergência das primeiras larvas nos capins. A ação do fungo foi avaliada no 35º, 38º, 41º, 48º, 55º e 61º dia pós-infestação, por meio da contagem de larvas recuperadas. Obteve-se controle de larvas do ácaro, que, nas avaliações realizadas entre o 35º e o 48º dia pós-infestação, variou entre 87% e 94%. As médias das contagens de estágios larvares do carrapato foram menores em todas as amostragens realizadas no capim-Tifton 85, indicando que houve efeito da pastagem na ação do fungo. A situação de aplicação influencia a atividade do fungo, com melhor resultado nas coletas realizadas entre o 41º e 55º dia após infestação em B. brizantha, e aplicação dos conídios logo após a emergência das primeiras larvas.The objective of this work was to evaluate the efficiency of Metarhizium anisopliae fungus against Boophilus microplus population in Brachiaria brizantha and Tifton 85 (Cynodon pastures, artificially infested with tick engorged females. Thirty plots of 1 m² each were randomly distributed in fifteen treated and fifteen control groups per type of grass, establishing five repetitions for each treatment. Pastures were infested with engorged tick females

  14. Compatibilidade de Metarhizium anisopliae (Metschnikoff Sorokin (Hypocreales: Clavicipitaceae com Chrysoperla externa (Hagen (Neuroptera: Chrysopidae

    Directory of Open Access Journals (Sweden)

    Eunice Claúdia Souza

    2015-12-01

    Abstract. The ecoagrossistemas several biological control agents acting simultane ously on different or the same target insect. The culture of cotton presents a rich complex of natural enemies associated with pests, among which stands out the predator Chrysoperla externa (Hagen and the entomopathogenic fungus Metarhizium anisopliae (Metschnikoff Sorokin. The objective of this study was to evaluate the compatibility of the fungus M. anisopliae on the biological aspects of the predator C. externa. The first study evaluated the development of C. externa when treated with the following suspensions of M. anisopliae: T1 water (control; T2- 105 conídios.mL-1 de M. anisopliae (M.a.; T3- 106 conídios.mL-1 de M.a.; T4- 107 conídios.mL-1 de M.a.;T5- 108 conídios.mL-1 de M.a.. The second assay measured the duration of the larval and pupal predator and larval mortality in each treatment. For this, C. externa eggswere individualized in Petri dishes and kept under laboratory conditions. When the larvae of C. externa hatched, whitefly nymphs were offered on leaf discs coming from cotton treated with: T1-water (control; T2 - 105 conídios.mL-1 de M.a. e T3 - 108 conídios.mL-1 de M.a.. The results suggest that the fungus M. anisopliae at doses of 105 e 108 conídios.mL-1 de M.a. not interfere with the biological aspects of the predator C. externa.

  15. LIQUID BIOFORMULATION OF METARHIZIUM ANISOPLIAE IS EFFECTIVE FOR THE MANAGEMENT OF COW PEA MOSAIC DISEASE

    Directory of Open Access Journals (Sweden)

    Sarodee Boruah

    2015-03-01

    Full Text Available Liquid bioformulation of Metarhizium anisopliae amended with oils and adjuvants was prepared. Six oils viz., sunflower, safflower, soybean, mustard, arachnid and coconut oils at three different concentrations, 0.1, 0.5 and 1.0 per cent and three adjuvants viz., glycerol @ 5, 8 and 10 per cent; tween-80 @ 0.02, 0.04, 0.06 and 1.0 per cent and arachnid oil @ 1, 5 and 10 per cent were tested to see their effect on growth and development of M. anisopliae in the bioformulation. Liquid bioformulation of M. anisopliae amended with Glycerol (10.0% + Sunflower oil (0.5% was found significantly effective which showed 84.33 per cent and 94.93 per cent higher surface area covered and biomass production respectively than the control. Efficacy of the bioformulation was tested against cow pea aphid, Aphis craccivora in pot as well as field condition. Liquid formulation of M. anisopliae supplemented with Glycerol (10.0% + Sunflower oil (0.5% was found to be significantly effective causing aphid mortality of 80 per cent at 30 days after spraying with protection of secondary spread of cow pea mosaic disease up to 100 per cent and 96.03 per cent in pot and field condition respectively. Spraying liquid formulation of M. anisopliae amended with Glycerol (10.0% + Sunflower oil (0.5% at 15 days interval for twice proved to be the best treatment with highest yield of 24.03 t/ha with a cost benefit ratio of 1:7.95 as compared to control (11.20 t/ha.

  16. A Field Experiment to Assess the Rate of Infestation in Honey Bee Populations of Two Metarhizium Anisopliae Isolates on Varroa Destructor (Acari: Mesostigmata

    Directory of Open Access Journals (Sweden)

    Khodadad Pirali-kheirabadi

    2013-03-01

    Full Text Available Background: The protective effect of two isolates of an entomopathogenic fungus, Metarhizium anisopliae (DEMI 002 and Iran 437C on the adult stage of Varroa destructor was evaluated in comparison with fluvalinate strips in the field.Methods: A total of 12 honey bee colonies were provided from an apiculture farm. The selected hives were divided into 4 groups (3 hives per group. The first group was the control, treated with distilled water. The other two groups were exposed to different fungi (M. anisopliae isolates DEMI 002 and Iran 437C and the last group was treated with one strip of fluvalinate per colony. The number of fallen mites was counted using sticky traps during a 6-day period, six days before and after treatments. A fungal suspension at a concentration of 5× 106 conidia/mL was sprayed onto the frames and the number of fallen mites was counted.Results: Metarhizium anisopliae DEMI 002 and Iran 437C isolates were as effective (i.e., caused as much mite fall as the fluvalinate strip in controlling bee colonies than no treatment.Conclusion: Both M. anisopliae isolates are promising candidates as agents in the control of Varroa mites under field conditions. Isolate DEMI 002 can be considered as a possible non-chemical biocontrol agent for controlling bee infestation with V. destructor in the field. In order to substantiate this hypothesis, tests are currently being performed using larger colonies and larger doses than tested in the present study in our beekeeping.

  17. Virulence of Mexican isolates of entomopathogenic fungi upon Rhipicephalus-Boophilus microplus larvae and the efficacy of conidia formulations to reduce larval tick density under field conditions

    Science.gov (United States)

    The first objective was laboratory evaluation of the virulence of 53 Mexican isolates of fungi against larvae of Rhipicephalus (Boophilus) microplus. Thirty three isolates of Metarhizium anisopliae var. anisopliae (Metschnickoff) Sorokin (Hypocreales: Clavicipitaceae) and 20 isolates of Isaria (Paec...

  18. Molecular genetics of secondary chemistry in Metarhizium fungi

    Science.gov (United States)

    As with many microbes, entomopathogenic fungi from the genus Metarhizium produce a plethora of small molecule metabolites, often referred to as secondary metabolites. Although these intriguing compounds are a conspicuous feature of the biology of the producing fungi, their roles in pathogenicity and...

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

    Directory of Open Access Journals (Sweden)

    Luz Christian

    1998-01-01

    Full Text Available 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.

  20. Biochemical basis of synergism between pathogenic fungus Metarhizium anisopliae and insecticide chlorantraniliprole in Locusta migratoria (Meyen)

    Science.gov (United States)

    Jia, Miao; Cao, Guangchun; Li, Yibo; Tu, Xiongbing; Wang, Guangjun; Nong, Xiangqun; Whitman, Douglas W.; Zhang, Zehua

    2016-01-01

    We challenged Locusta migratoria (Meyen) grasshoppers with simultaneous doses of both the insecticide chlorantraniliprole and the fungal pathogen, Metarhizium anisopliae. Our results showed synergistic and antagonistic effects on host mortality and enzyme activities. To elucidate the biochemical mechanisms that underlie detoxification and pathogen-immune responses in insects, we monitored the activities of 10 enzymes. After administration of insecticide and fungus, activities of glutathione-S-transferase (GST), general esterases (ESTs) and phenol oxidase (PO) decreased in the insect during the initial time period, whereas those of aryl acylamidase (AA) and chitinase (CHI) increased during the initial period and that of acetylcholinesterase (AChE) increased during a later time period. Activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) decreased at a later time period post treatment. Interestingly, treatment with chlorantraniliprole and M. anisopliae relieved the convulsions that normally accompany M. anisopliae infection. We speculate that locust mortality increased as a result of synergism via a mechanism related to Ca2+ disruption in the host. Our study illuminates the biochemical mechanisms involved in insect immunity to xenobiotics and pathogens as well as the mechanisms by which these factors disrupt host homeostasis and induce death. We expect this knowledge to lead to more effective pest control. PMID:27328936

  1. Catalase overexpression reduces the germination time and increases the pathogenicity of the fungus Metarhizium anisopliae.

    Science.gov (United States)

    Morales Hernandez, Claudia Erika; Padilla Guerrero, Israel Enrique; Gonzalez Hernandez, Gloria Angelica; Salazar Solis, Eduardo; Torres Guzman, Juan Carlos

    2010-07-01

    Catalases and peroxidases are the most important enzymes that degrade hydrogen peroxide into water and oxygen. These enzymes and superoxide dismutase are the first lines of cell defense against reactive oxygen species. Metarhizium anisopliae displays an increase in catalase-peroxidase activity during germination and growth. To determine the importance of catalase during the invasion process of M. anisopliae, we isolated the cat1 gene. cat1 cDNA expression in Escherichia coli and the subsequent purification of the protein confirmed that the cat1 gene codes for a monofunctional catalase. Expression analysis of this gene by RT-PCR from RNA isolated from fungus grown in liquid cultures showed a decrease in the expression level of the cat1 gene during germination and an increase during mycelium growth. The expression of this gene in the fungus during the infection process of the larvae of Plutella xylostella also showed a significant increase during invasive growth. Transgenic strains overexpressing the cat1 gene had twice the catalase activity of the wild-type strain. This increase in catalase activity was accompanied by a higher level of resistance to exogenous hydrogen peroxide and a reduction in the germination time. This improvement was also observed during the infection of P. xylostella larvae. M. anisopliae transgenic strains overexpressing the cat1 gene grew and spread faster in the soft tissue of the insect, reducing the time to death of the insect by 25% and the dose required to kill 50% of the population 14-fold.

  2. Route of infection and hematological effect of Metarhizium anisopliae (Metsch.) Sorokin on Dysdercus cingulatus (Fab.) adult.

    Science.gov (United States)

    Sahayaraj, Kitherian; Borgio, Jesu Francis; Lucini, Luigi

    2014-01-01

    The primary objective of this work was to identify, under laboratory conditions, the route of infection and hemogram of Dysdercus cingulatus (Fab.) adults by Metarhizium anisopliae. The infection process in D. cingulatus by M. anisopliae involved the conidia adherence to the host cuticle and germination after 24 h post-infection, accompanied by falling of bristles. The subsequent step, within 24-48 h post-infection, comprised penetration of fungus through spiracles, root of bristles, hemolymph, and the three dorsal sacs. Subsequently, within 72-96 h post-infection, the fungus penetrated into trachea and sacs, then emerged on cuticular surface and was found to be maximum in hemolymph. A great decrease in hemocytes count was observed within 96 h from infection. The hemosomic index (HSI) decreased gradually as the incubation period increased. As far as we know, this is the first study to know the mechanism of action of M. anisopliae to D. cingulatus.

  3. Proteins differentially expressed in conidia and mycelia of the entomopathogenic fungus Metarhizium anisopliae sensu stricto.

    Science.gov (United States)

    Su, Yubin; Guo, Qingfeng; Tu, Jie; Li, Xiaoxia; Meng, Lixue; Cao, Liping; Dong, Dong; Qiu, Junzhi; Guan, Xiong

    2013-07-01

    Metarhizium anisopliae is a well-characterized entomopathogenic fungus that attacks a variety of insects. Its conidia are involved in its propagation and also in its infection of host insects. To investigate the protein expression profiles and to identify the proteins related to development and pathogenesis, we performed a comparative proteomic analysis of the conidia and mycelia of an M. anisopliae strain (Ma1291). The analysis used 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. We detected 898 ± 37 protein spots in conidia and 1072 ± 24 in mycelia of strain Ma1291. A comparison of the 2 protein-expression profiles indicated that only 28% of protein spots were common to both developmental stages. Finally, we identified 30 proteins (19 from conidia and 11 from mycelia). The identified proteins exclusive to conidia were those involved in protective processes, appressorium formation, and degradation of the host cuticle (protease PR1H). The identified proteins exclusive to mycelia included major proteins participating in biosynthetic and energy metabolism, such as UTP-glucose-1-phosphate uridylyltransferase and heat shock protein 70. This research provides the first proteomic analysis of different developmental stages of M. anisopliae, and the results should facilitate clarification of the molecular basis of these epigenetic variations.

  4. Effects of Metarhizium anisopliae conidia mixed with soil against the eggs of Aedes aegypti.

    Science.gov (United States)

    Leles, Renan Nunes; D'Alessandro, Walmirton Bezerra; Luz, Christian

    2012-04-01

    The effectiveness of Metarhizium anisopliae IP 46 conidia mixed with soil was tested against Aedes aegypti eggs. Mycelium and new conidia developed first on eggs between 4.8 and 15 days respectively after incubation of fungus-treated soils at 3.3 × 10(3) up to 3.3 × 10(5) conidia/g soil at 25°C and relative humidities close to saturation. After 15-day incubation, 53.3% of the eggs exposed to soil with 3.3 × 10(5) conidia/g showed external development of mycelium and conidia. Fungus-inoculated soils (but not untreated controls) showed some mycelial growth and sporulation apart from the eggs. Some eggs on treated soils hatched; those larvae died and eventually showed fungal development on their bodies. The cumulative relative eclosion of larvae after submersion of treated eggs in water decreased from 52.2% at 3.3 × 10(3) conidia/g to 25.3% at 3.3 × 10(5) conidia/g. These findings clearly showed that A. aegypti eggs can be infected by M. anisopliae when deposited on fungus-contaminated soil. The effectiveness of M. anisopliae against gravid females, larvae, and also eggs of A. aegypti underscored the possible usefulness of this fungus as a mycoinsecticide, whether naturally occurring or artificially applied, in the breeding sites of this mosquito.

  5. Potential nontarget effects of Metarhizium anisopliae (Deuteromycetes) used for biological control of ticks (Acari: Ixodidae)

    Science.gov (United States)

    Ginsberg, Howard S.; LeBrun, Roger A.; Heyer, Klaus; Zhioua, Elyes

    2002-01-01

    The potential for nontarget effects of the entomopathogenic fungus Metarhizium anisopliae (Metschnikoff) Sorokin, when used for biological control of ticks, was assessed in laboratory trials. Fungal pathogenicity was studied against convergent ladybird beetles, Hippodamia convergens Guérin-Méneville, house crickets, Acheta domesticus (L.), and the milkweed bugs Oncopeltus fasciatus (Dallas). Fungal spores applied with a spray tower produced significant mortality in H. convergens and A. domesticus, but effects on O. fasciatus were marginal. Placing treated insects with untreated individuals resulted in mortality from horizontal transmission to untreated beetles and crickets, but not milkweed bugs. Spread of fungal infection in the beetles resulted in mortality on days 4–10 after treatment, while in crickets mortality was on day 2 after treatment, suggesting different levels of pathogenicity and possibly different modes of transmission. Therefore, M. anisopliae varies in pathogenicity to different insects. Inundative applications can potentially affect nontarget species, but M. anisopliae is already widely distributed in North America, so applications for tick control generally would not introduce a novel pathogen into the environment. Pathogenicity in lab trials does not, by itself, demonstrate activity under natural conditions, so field trials are needed to confirm these results and to assess methods to minimize nontarget exposure.

  6. Biochemical basis of synergism between pathogenic fungus Metarhizium anisopliae and insecticide chlorantraniliprole in Locusta migratoria (Meyen).

    Science.gov (United States)

    Jia, Miao; Cao, Guangchun; Li, Yibo; Tu, Xiongbing; Wang, Guangjun; Nong, Xiangqun; Whitman, Douglas W; Zhang, Zehua

    2016-06-22

    We challenged Locusta migratoria (Meyen) grasshoppers with simultaneous doses of both the insecticide chlorantraniliprole and the fungal pathogen, Metarhizium anisopliae. Our results showed synergistic and antagonistic effects on host mortality and enzyme activities. To elucidate the biochemical mechanisms that underlie detoxification and pathogen-immune responses in insects, we monitored the activities of 10 enzymes. After administration of insecticide and fungus, activities of glutathione-S-transferase (GST), general esterases (ESTs) and phenol oxidase (PO) decreased in the insect during the initial time period, whereas those of aryl acylamidase (AA) and chitinase (CHI) increased during the initial period and that of acetylcholinesterase (AChE) increased during a later time period. Activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) decreased at a later time period post treatment. Interestingly, treatment with chlorantraniliprole and M. anisopliae relieved the convulsions that normally accompany M. anisopliae infection. We speculate that locust mortality increased as a result of synergism via a mechanism related to Ca(2+) disruption in the host. Our study illuminates the biochemical mechanisms involved in insect immunity to xenobiotics and pathogens as well as the mechanisms by which these factors disrupt host homeostasis and induce death. We expect this knowledge to lead to more effective pest control.

  7. Biological control of Ixodes ricinus larvae and nymphs with Metarhizium anisopliae blastospores.

    Science.gov (United States)

    Wassermann, Marion; Selzer, Philipp; Steidle, Johannes L M; Mackenstedt, Ute

    2016-07-01

    The entomopathogenic fungus Metarhizium anisopliae is used as a biological pest control agent against various arthropod species, including ticks. However, the efficacy depends on tick species, tick stage and fungus strain. We studied the effect of M. anisopliae on engorged larvae and nymphs of Ixodes ricinus, the most abundant tick species in Europe, under laboratory and semi-field conditions. A significant reduction of engorged larvae and nymphs could be shown under laboratory as well as under semi-field conditions. Only 3.5% of the larvae treated in the lab and only 18.5% kept under semi-field conditions were able to develop into nymphs compared to the recovered nymphs of the control groups, which were regarded as 100%. Only 7.1% of nymphs were recovered as adult ticks after fungal treatment under semi-field conditions compared to the control (100%). The efficacy of blastospores of M. anisopliae against engorged larvae and nymphs of I. ricinus under semi-field conditions was demonstrated in this study, showing their high potential as a biological control agent of ticks. Further studies will have to investigate the effect of this agent against other stages of I. ricinus as well as other tick species before its value as a biological control agent against ticks can be fully assessed.

  8. The antileukaemic cell cycle regulatory activities of swainsonine purified from Metarhizium anisopliae fermentation broth.

    Science.gov (United States)

    Singh, Digar; Kaur, Gurvinder

    2014-01-01

    Swainsonine is a Metarhizium secondary metabolite known differentially for its specific mannosidase inhibitory, toxic and therapeutic activities. Here, the standard and purified swainsonine from Metarhizium anisopliae fermentation broth were comparatively evaluated for their in situ antileukaemic activities in human promyelocytic cell line, HL-60. Both the standard (IC50 = 6.96 μM) and purified (IC50 = 9.50 μM) compounds inhibited the leukaemic cell proliferation without inflicting cell membrane disruption at 48 h of post-treatment incubation. The DNA cell cycle analysis showed approximately 48.81% and 60.72% of the treated cells arrested in the synthetic phase (S-phase) at 36 and 48 h, respectively, upon treatment with IC50 concentration of the purified swainsonine. However, only 29.62% of cells were arrested in S-phase with standard swainsonine at 48 h, suggesting the comprehensive action of certain other metabolites sharing the similar paradigm of antiproliferative properties in Metarhizium broth extract.

  9. Efficacy of enthomopathogenic fungus Metarhizium anisopliae against Tuta absoluta (Lepidoptera: Gelechiidae).

    Science.gov (United States)

    Contreras, J; Mendoza, J E; Martínez-Aguirre, M R; García-Vidal, L; Izquierdo, J; Bielza, P

    2014-02-01

    The tomato borer Tuta absoluta Meyrick is a serious tomato pest that has lately undergone a rapid expansion, causing severe crop losses. An integrated management is required to control this insect, within which biological control is now beginning to play a key role. In this regard, the effectiveness of a liquid formulation based on strains of the fungus Metarhizium anisopliae variety anisopliae (Metschnikoff) Sorokin (4.46 x 10(9) viable conidia per milliliter), applied together with irrigation water, has been evaluated by laboratory tests on different populations of T. absoluta. A bioassay method has been developed to test the efficacy of the product. The technique chosen has been validated and the different studied populations have been typified according to their susceptibility, determining the baseline susceptibility of the pest to the fungus. The results revealed a complete efficacy of M. anisopliae against pupae of T. absoluta at the recommended label rate (5.58 x 10(9) viable conidia per liter) for the populations assayed. Moreover, a notably lower dose was also sufficiently effective to control the tomato borer populations because values of LC90 lower than 3 x 10(9) viable conidia per liter were obtained unfailingly. The most sensitive populations were those collected in Almeria and Nijar, Spain, with LC50 values of 0.21 and 0.22 x 10(9) viable conidia per liter(-1), respectively. The average value obtained for LC50 was 0.34 x 10(9) viable conidia per liter(-1) and 2 x 10(9) for LC90. These results show the potential of M. anisopliae to control pupae of the tomato borer in integrated pest management programs.

  10. Laboratory Bioassay of Iranian Isolates of Entomopathogenic Fungus Metarhizium anisopliae (Metsch. Sorokin (Ascomycota: Hypocreales against two Species of Storage Pest

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    Hamid Sakenin Chelav

    2013-03-01

    Full Text Available The susceptibility of adults of Tribolium castaneum Herbst (Coleoptera: Tenebrionidae and Oryzaephilus surinamensis L. (Coleoptera: Silvanidae to three Iranian Isolates of the entomopathogenic fungi Metarhizium anisopliae (Metsch. Sorokin was evaluated through bioassays with direct immersion under laboratory conditions. For each isolates, five aqueous suspensions were prepared in a logarithmic series in Tween 80 (0.05% v/v. Results showed that adult of both species were susceptible to all isolates of M. anisopliae. For all three isolates, mortality percentage of the two species increased with increasing conidial concentration and significant difference was observed between concentrations. The corrected cumulative mortality of adult of O. surinamensis 10 days after immersion ranged from 12.38 to 85.84%, 18.6 to 62.83% and 10.63 to 77.87% for different concentrations of DEMI001, IRAN 715C and IRAN 1018C, respectively. These amounts for T. castaneum varied from 31.07 to 74.78%, 26.02 to 75.61% and 23.33 to 89.99% for different concentrations of DEMI001, IRAN 715C and IRAN 1018C, respectively. The parameters of probit analysis demonstrated non-overlap of 95% confidence limits of LC50 and LC95 and significant difference was observed among three isolates tested against each insect. The lowest and the highest LC50 and LC95 values were observed in the isolates DEMI001 for O. surinamensis (3/1×105 and 1/5×108 and IRAN 715C for T. castaneum (6.2×108 and 6.9×1014, respectively. This observation highlights the importance and need of screening for more virulent isolates against storage pests for use in the management of these pests.

  11. DNA polymorphism and total protein in mutants of Metarhizium anisopliae var. Anisopliae (Metsch. Sorokin strain E9 Polimorfismo de DNA e proteína total em mutantes da linhagem E9 de Metarhizium anisopliae var. anisopliae (Metsch. Sorokin

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    Laurineide Lopes de Carvalho Freire

    2001-06-01

    Full Text Available Five mutants (MaE10, MaE27, MaE24, MaE41 e MaE49 of Metarhizium anisopliae wild strain E9 were analysed for DNA profile through the RAPD technique and for changes in total protein content by spectrophotometry, polyacrylamide gel electrophoresis and densitometry. The pattern of RAPD markers showed genetic polymorphism among the strains: out of twenty primers seven were selected, producing 113 bands. Forty seven bands were present in all strains (41.6% of monomorphic bands and 66 showed polymorphism (58.4%. The mean coefficient of similarity among all strains was 0.75 (75%. The total protein content varied, staining in the interval of 6.0-8.0 µg/µl. The electrophoresis analysis, through zymogram and protein fraction profiles by densitometry, allowed the observation of seven bands for the wild strain E9 and five bands for the mutants MaE10, MaE27, MaE34, MaE41 and MaE49, evidence of variations in µg% among protein fractions. The RAPD technique was very sensitive to detect genetic differences between the wild type and the mutants obtained through gamma radiation. The total protein analysis also showed changes in quantity and pattern of bands after electrophoresis in the mutants compared to the wild type.Foram analisados cinco mutantes MaE (MaE10, MaE27, MaE34, MaE41 e MaE49 da linhagem selvagem E9 de Metarhizium anisopliae var. anisopliae quanto ao perfil de DNA pela técnica de RAPD e também quanto ao conteúdo de proteína total por espectrometria e eletroforese em gel de poliacrilamida e densitometria. O padrão de marcadores de RAPD evidenciou polimorfismo nas amostras; dos 20 primers testados foram selecionados 7 que geraram 113 bandas. Deste total, 47 estavam presentes em todas as amostras (41.6% de bandas monomórficas e 66 mostraram polimorfismo (58.4%. O coeficiente médio de similaridade foi de 75%. O conteúdo de proteína total variou de 6 a 8 µg/µl. O zimograma e perfís das frações de proteínas obtidos por densitometria

  12. The rhizosphere-competent entomopathogen Metarhizium anisopliae expresses a specific subset of genes in plant root exudate.

    Science.gov (United States)

    Pava-Ripoll, Monica; Angelini, Claudia; Fang, Weiguo; Wang, Sibao; Posada, Francisco J; St Leger, Raymond

    2011-01-01

    Metarhizium anisopliae and Beauveria bassiana are ubiquitous insect pathogens and possible plant symbionts, as some strains are endophytic or colonize the rhizosphere. We evaluated 11 strains of M. anisopliae and B. bassiana, and two soil saprophytes (the non-rhizospheric Aspergillus niger and the rhizosphere-competent Trichoderma harzianum) for their ability to germinate in bean root exudates (REs). Our results showed that some generalist strains of M. anisopliae were as good at germinating in RE as T. harzianum, although germination rates of the specialized acridid pathogen Metarhizium acridum and the B. bassiana strains were significantly lower. At RE concentrations of anisopliae strain ARSEF 2575 showed higher germination rates than T. harzianum. Microarrays showed that strain 2575 upregulated 29 genes over a 12 h period in RE. A similar number of genes (21) were downregulated. Upregulated genes were involved in carbohydrate metabolism, lipid metabolism, cofactors and vitamins, energy metabolism, proteolysis, extracellular matrix/cell wall proteins, transport proteins, DNA synthesis, the sexual cycle and stress response. However, 41.3% of the upregulated genes were hypothetical or orphan sequences, indicating that many previously uncharacterized genes have functions related to saprophytic survival. Genes upregulated in response to RE included the subtilisin Pr1A, which is also involved in pathogenicity to insects. However, the upregulated Mad2 adhesin specifically mediates adhesion to plant surfaces, demonstrating that M. anisopliae has genes for rhizosphere competence that are induced by RE.

  13. Impact of moisture on survival of Aedes aegypti eggs and ovicidal activity of Metarhizium anisopliae under laboratory conditions.

    Science.gov (United States)

    Luz, C; Tai, M H H; Santos, A H; Silva, H H G

    2008-03-01

    The effect of relative humidity (43%, 75%, 86% and > 98%) on Aedes aegypti eggs treated with Metarhizium anisopliae or water only was tested for up to a six months exposure at 25 degrees C. Survival of larvae inside eggs was clearly affected by the lowest humidity (43%) tested, and eclosion diminished at all humidities after increasing periods of exposure. M. anisopliae showed to have a strong ovicidal activity only at humidity close to saturation. No difference of activity was found between conidia and hyphal bodies tested. This fungus affected larvae inside eggs and has potential as a control agent of this important vector in breeding sites with high moisture.

  14. Impact of moisture on survival of Aedes aegypti eggs and ovicidal activity of Metarhizium anisopliae under laboratory conditions

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    C Luz

    2008-03-01

    Full Text Available The effect of relative humidity (43%, 75%, 86% and > 98% on Aedes aegypti eggs treated with Metarhizium anisopliae or water only was tested for up to a six months exposure at 25ºC. Survival of larvae inside eggs was clearly affected by the lowest humidity (43% tested, and eclosion diminished at all humidities after increasing periods of exposure. M. anisopliae showed to have a strong ovicidal activity only at humidity close to saturation. No difference of activity was found between conidia and hyphal bodies tested. This fungus affected larvae inside eggs and has potential as a control agent of this important vector in breeding sites with high moisture.

  15. New antibacterial isocoumarin glycosides from a wetland soil derived fungal strain Metarhizium anisopliae.

    Science.gov (United States)

    Tian, Jie-Feng; Li, Peng-Ju; Li, Xiao-Xia; Sun, Ping-Hua; Gao, Hao; Liu, Xing-Zhong; Huang, Peng; Tang, Jin-Shan; Yao, Xin-Sheng

    2016-03-01

    Eight new isocoumarin glycosides (1-8) were obtained from the solid culture of the wetland soil-derived fungus Metarhizium anisopliae (No. DTH12-10). Their chemical structures were elucidated by analyses of HR ESI-TOF MS, (1)H, (13)C NMR, (1)H-(1)H COSY, HSQC, and HMBC spectra. The absolute configurations were determined by single crystal X-ray diffraction, circular dichroism (CD) spectrum, and chemical derivatization methods. In addition, inhibition of the biofilm formation and the secretion of virulence factor of the new isocoumarin glycosides against Pseudomonas aeruginosa strain PAOA (clinical isolates) were evaluated. The result revealed that compound 1 showed antibacterial activity comparable with (Z)-4-bromo-5-(bromomethylene)-2(5H)-furanone (BF).

  16. Estimates of genetic parameters related to chitinase production by the entomopathogenic fungus Metarhizium anisopliae

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    Gilberto U.L. Braga

    1998-06-01

    Full Text Available Chitinolytic activity and dry mass production were determined in culture filtrates from 17 Metarhizium anisopliae strains grown in liquid medium containing chitin as the only carbon source. The objectives were to estimate parameters such as genetic variance among strains, heritability and expected gain from selection, as well as correlations between tested traits. Wide genotypic variability was observed among strains in chitinolytic activity, permitting the exploitation of this property in selection. The high heritability suggests that progress can be made through phenotypic selection. The genotypic correlation coefficient between dry mass production and chitinolytic activity detected in the filtrates was negative (= - 0.588. One of the isolates was also investigated for variation in the two traits as a function of culture growth time. The results showed an increase in enzyme activity up to the 8th (and last day of the experiment and a decrease in dry mass from the 4th day on.Foi determinada a atividade quitinolítica apresentada pelos filtrados de culturas de 17 genótipos de Metarhizium anisopliae crescidos em meio líquido, tendo quitina como única fonte de carbono. O objetivo foi a estimativa de parâmetros como a variância genética, a herdabilidade e o ganho esperado na seleção. Uma grande variabilidade genotípica foi verificada na atividade quitinolítica, permitindo sua exploração no melhoramento. Os altos coeficientes de herdabilidade permitem esperar um grande progresso na seleção fenotípica. Para uma das linhagens foi também determinada a variação na atividade enzimática, em função do tempo de crescimento das culturas. Os resultados mostraram um aumento na atividade enzimática até o oitavo dia da avaliação.

  17. Pathogenicity of the entomopathogenic fungus Metarhizium anisopliae var. major on different stages of the sunn pest Eurygaster integriceps.

    Science.gov (United States)

    Sedighi, Neda; Abbasipour, Habib; Askary, Hassan; Gorjan, Aziz Sheikhi; Karimi, Jaber

    2013-01-01

    The sunn pest, Eurygaster integriceps Puton (Hemiptera: Scutelleridae), is the most important insect pest of wheat and barley. The population management of this pest is of major concern to wheat producers. One of the potential control strategies is to use entomopathogenic fungi. This study evaluates the pathogenicity of the fungus Metarhizium anisopliae var. major (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae) on the sunn pest, E. integriceps. Five concentrations of the fungus were utilized, ranging from 1×10(4) to 1×10(8) conidia/mL, accompanied by controls. Fifth instar nymphs and adults (a migratory summer population and a diapausing population) previously exposed to fungi were sown to isolate the fungi, and the growth parameters were analyzed. A direct spray technique was used to expose the isolates to the E. integriceps. The experiment was repeated four times, and mortalities of the insects for all treatments were recorded daily. The results showed that the mortality of infected nymphs was significantly higher than the mortality of control nymphs. Also, the longevity of infected adults was lower than the controls. The results also showed that diapausing adults of the sunn pest were much more susceptible to infection than the summer adults. Estimated LC50 values for the M14 isolate were 1.4 × 10(6), 1.4 ×10 (5) , and 2.3 × 10(3) spores/mL against the aestivation population, the diapausing population, and 5(th) instar nymphs, respectively. Estimated LT50 values using 10(8) spores/mL of the Mm isolate on the aestivation and diapausing populations were 11.9 and 5.11 days, respectively. The results demonstrated that M. anisoplaie was effective on all of stages of E. integriceps. In addition, the nymphal stage was more susceptible than the adults.

  18. Infectivity of Metarhizium anisopliae (Hypocreales: Clavicipitaceae) to Phlebotomus papatasi (Diptera: Psychodidae) under laboratory conditions.

    Science.gov (United States)

    Zayed, Alia; Soliman, Mustafa M; El-Shazly, Mohamed M

    2013-07-01

    Susceptibility of Phlebotomus papatasi Scopoli (Diptera: Psychodidae) larvae to the entomopathogenic fungus Metarhizium anisopliae (Metschinkoff) Sorokin (Ma79) (Hypocreales: Clavicipitaceae) was evaluated at two different temperatures. The ability of the fungus to reinfect healthy sand flies was followed up for approximately 20 wk and the effect of in vivo repassage on the enhancement of its virulence was assessed. The fungus reduced the adult emergence at 26 +/- 1 degrees C when applied to larval diet. Six spore concentrations were used in the bioassays ranging from 1 x 10(6) to 5 x 10(8) spores/ml. Mortality decreased significantly when the temperature was raised to 31 +/- 1 degrees C at all tested concentrations. Fungus-treated vials were assayed against sand fly larvae at different time lapses without additional reapplication of the fungus in the media to determine whether the level of inocula persisting in the media was sufficient to reinfect healthy sand flies. Twenty weeks postapplication, there were still enough infectious propagules of Ma79 to infect 40% of P. papatasi larvae. A comparison between the infectivity of 10 subsequent in vitro cultures and the host-passed inocula of the fungus against sand fly larvae was conducted. Mortalities of P. papatasi larvae changed significantly when exposed to inocula passed through different insects. Presented data can provide vector control decision makers and end users with fundamental information for the introduction and application of M. anisopliae as an effective control agent against the main cutaneous leishmaniasis old-world vector P. papatasi.

  19. Expression of chitinase genes of Metarhizium anisopliae isolates in lepidopteran pests and on synthetic media.

    Science.gov (United States)

    Bhanu Prakash, G V S; Padmaja, V; Jami, Sravan Kumar; Kirti, P B

    2012-12-01

    Pathogenecity of the well characterized entomopathogenic fungus Metarhizium anisopliae used for biocontrol of a wide range of insect pests secretes hydrolytic enzymes that degrade the host cuticle. The chitinolytic activity of high and low virulent isolates of M. anisopliae was assayed on minimal medium (MM) + colloidal chitin and MM supplemented with insect cuticles. Ex- pression pattern of four chitinase genes (chitinase (chi), chi 1, chi 2, chi 3) was profiled during pathogenic stages of the entomopathogen under in vitro and in vivo conditions. Reverse-transcription polymerase chain reaction (RT-PCR) analysis confirmed that chitinase cDNAs were expressed during the germination of fungus under nutrient-deprived conditions. RT-PCR analysis performed for the four chitinase genes on the two insect hosts Spodoptera litura and Helicoverpa armigera at six developmental stages of the pathogen displayed up-regulation in S. litura at mycosed and conidiated condition while with H. armigera there was expression only after 48 h of incubation. Differential expression of chi, chi 1 and chi 2 genes in vitro (nitrogen rich and nitrogen limiting media) and in vivo (live insect hosts S. litura and H. armigera) implicate the role of substrate differences in pathogenesis.

  20. New insights on the effectiveness of Metarhizium anisopliae formulation and application against Aedes aegypti eggs.

    Science.gov (United States)

    Sousa, N A; Lobo, L S; Rodrigues, J; Luz, C

    2013-09-01

    Increasing needs for innovative control tools against the dengue vector Aedes aegypti have prompted investigations into the development of specific mycoinsecticides. The entomopathogenic fungus Metarhizium anisopliae attacks both larval and adult stages, but its ovicidal activity against A. aegypti is still little explored. This study reports important findings about the effectiveness of conidia formulated in water and oil-in-water emulsions and of direct and indirect application techniques against A. aegypti eggs. The ovicidal activity of M. anisopliae increased with higher conidial concentrations regardless of the application technique, and larvae elimination concentrations were lowest with oil-in-water-formulated conidia (LEC50 ≤ 4·8 × 10(3) conidia cm(-2) and LEC90 ≤ 1·9 × 10(5) conidia cm(-2), respectively). Conidia eventually stimulated larval eclosion. Consequently, the indirect application of oil-based fungal formulations onto substrates where oviposition will later occur appears to be a more efficient means to infect those eggs than the direct fungal application to previously deposited eggs.

  1. Evaluating Toxicity of Extracted Destruxin from Metarhizium Anisopliae Against Citrus Leafminer, Phyllocnistis citrella

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    M. M. Sharif

    2010-01-01

    Full Text Available Problem statement: The fungus, Metarhizium anisopliae (Metschnikoff Sorokin produces some cyclic peptide toxins, destruxins, which exhibit a variety of insecticidal actions. Citrus leafminer, Phyllocnistis citrella Stainton, is a major pest of young citrus trees; therefore its effective and safe control is a important issue. Approach: Five isolates of M. anisopliae were inoculated in Potato Dextrose Broth (PDB mediums. Destruxin was extracted by adding chloroform. Citrus leafminer was bioassayed by using the leaves containing early stages larvae and the data were recorded after 1, 2, 3 and 4 days after treatment. Results: The range of mortality was between 30-65% based on the endpoint data. The minimum of three days to achieve 50% mortality was proved by probit analysis of time-mortality responses. The 10, 15 and 20 fold dilutions of extracted destruxins from isolates cause 17.2-48.5, 9.4-37.5 and 0-26.5%, respectively. Conclusion: Bioassay of destruxin by using citrus leafminer larvae shows its acceptable effect of destruxin. By considering biology of this species and calculated LT50s, destruxin seems to be an effectiveness component of IPM programs. The 10, 15 and 20 fold dilutions of extracted destruxin have a reduced amount of mortality, but in some cases it can balance the expensive production process of microbial agents and their efficacy.

  2. Horizontal Transmission of the Entomopathogen Fungus Metarhizium anisopliae in Microcerotermes diversus Groups.

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    Cheraghi, Amir; Habibpour, Behzad; Mossadegh, Mohammad Saied; Sharififard, Mona

    2012-08-08

    An experiment was carried out in order to investigate fungal conidia transmission of Metarhizium anisopliae (Metschnikoff) Sorokin from vector (donor) to healthy Microcerotermes diversus Silvestri (Iso.: Termitidae) and determine the best donor/concentration ratio for transmission. After preliminary trials, concentrations of 3.1 × 10⁴, 3.9 × 10⁵, 3.2 × 10⁶ and 3.5 × 10(8) conidia mL(-1) were selected for testing. The experiment was performed at three donor : Recipient ratios of 10, 30 and 50%. The highest mortality of recipient workers was observed after 14 days at the concentration of 3.5 × 10⁸ conidia mL(-1) and donor ratio of 50%. The mortality of recipient workers was less than 20% at all concentrations at a donor ratio of 10%. Our observations indicate social behavior of M. diversus, such as grooming, can be effective in promoting epizootic outbreaks in a colony. While the current results suggest good potential for efficacy, the use of M. anisopliae as a component of integrated pest management of M. diversus still needs to be proven under field conditions.

  3. Isolados de Metarhizium anisopliae (Metchnikoff Sorokin (Hypocreales: Clavicipitaceae para controle de Diatraea saccharalis Fabricius (Lepidoptera: Crambidae

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    Eduardo Hayashida

    2014-04-01

    Abstract. The selection of entomopathogenic fungus that is a necessary to know the efficiency of different isolates and one can choose the most suitable for use in biological control programs. This study aimed to evaluate the efficiency of the isolates of Metarhizium anisopliae (Metchnikoff Sorokin (UFGD 03, UFGD 05, UFGD UFGD 07 and UFGD 22, obtained in Mato Grosso do Sul on the Diatraea saccharalis Fabricius. The experimental trial consisted of a completely randomized design with five treatments and 10 repetitions with 5 caterpillars standardized size in each repetition. In general, M. anisopliae affects the life cycle of D. saccharalis and development of this pest is affected by the fungus. The percentage of caterpillars that pupae ranged from 64.00% to 88.60%. Treatment with isolated UFGD 03 had a higher percentage of dead pupae (68.00% compared to other isolates tested and compared to the control value (11.00%. The adult emergence of D. saccharalis was 87.02% in the control. In the treatments with the fungus, there was a variation of 35.20%, 38.00%, 40.00%, 52.80% provided by isolates UFGD 22, UFGD 03, UFGD 05 and UFGD 07, respectively.

  4. Compatibility of conventional agrochemicals used in rice crops with the entomopathogenic fungus Metarhizium anisopliae

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    Rodrigo Alves da Silva

    2013-06-01

    Full Text Available The toxicological impact of chemical pesticides on fungal entomopathogens and their use in tank-mixing can be directly measured through in vitro compatibility tests. This study reports the in vitro toxicity of eight insecticides, four fungicides and five herbicides in the conidial germination, vegetative growth and conidiation of Metarhizium anisopliae (strain CG 168. A conidial suspension containing the pesticide at recommended field dosage was subjected to constant agitation in a rotary shaker for 3h to simulate a tank mixing. Then, aliquots of each suspension were used to determine conidial germination, vegetative growth and conidiation on potato dextrose agar (PDA. The fungicides difenoconazole (69 mL ha-1, propiconazole (75 mL ha-1, trifloxystrobin (313 g ha-1 and azoxystrobin (56 mL ha-1 were the most harmful products to all biological stages of M. anisopliae and they should not be applied together with this fungus in tank mixing. The insecticides exhibited the least degree of toxicity to this fungal pathogen, whereas the herbicides had the greatest impact on mycelial growth. The agrochemicals compatible with M. anisopliae were the insecticides methyl parathion (240 mL ha-1, thiamethoxam (31 g ha-1, and lambda-cyhalothrin (6.3 mL ha-1 and the herbicides glyphosate (1560 mL ha-1, bentazon (720 mL ha-1, and imazapic+ imazapyr (84 g ha-1. The compatible pesticides could be simultaneously used with this bio-control agent for integrated pest management in rice production systems.

  5. Correlation between Pr1 and Pr2 gene content and virulence in Metarhizium anisopliae strains.

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    Rosas-García, Ninfa M; Avalos-de-León, Osvaldo; Villegas-Mendoza, Jesús M; Mireles-Martínez, Maribel; Barboza-Corona, J E; Castañeda-Ramírez, J C

    2014-11-28

    Metarhizium anisopliae is a widely studied model to understand the virulence factors that participate in pathogenicity. Proteases such as subtilisin-like enzymes (Pr1) and trypsin-like enzymes (Pr2) are considered important factors for insect cuticle degradation. In four M. anisopliae strains (798, 6342, 6345, and 6347), the presence of pr1 and pr2 genes, as well as the enzymatic activity of these genes, was correlated with their virulence against two different insect pests. The 11 pr1 genes (A, B, C, D, E, F, G, H, I, J, and K) and pr2 gene were found in all strains. The activity of individual Pr1 and Pr2 proteases exhibited variation in time (24, 48, 72, and 96 h) and in the presence or absence of chitin as the inductor. The highest Pr1 enzymatic activity was shown by strain 798 at 48 h with chitin. The highest Pr2 enzymatic activity was exhibited by the 6342 and 6347 strains, both grown with chitin at 24 and 48 h, respectively. Highest mortality on S. exigua was caused by strain 6342 at 48 h, and strains 6342, 6345, and 6347 caused the highest mortality 7 days later. Mortality on Prosapia reached 30% without variation. The presence of subtilisin and trypsin genes and the activity of these proteases in M. anisopliae strains cannot be associated with the virulence against the two insect pests. Probably, subtilisin and trypsin enzyme production is not a vital factor for pathogenicity, but its contribution is important to the pathogenicity process.

  6. Selection of indigenous isolates of entomopathogenic soil fungus Metarhizium anisopliae under laboratory conditions.

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    Skalický, Aleš; Bohatá, Andrea; Šimková, Jana; Osborne, Lance S; Landa, Zdeněk

    2014-07-01

    Eight native isolates of the entomopathogenic fungus Metarhizium anisopliae (Metschnikoff) Sorokin were obtained by monitoring soils cultivated in a conventional manner. These isolates were compared in three areas: (a) conidial germination, (b) radial growth and sporulation and (c) ability of conidia to infect Tenebrio molitor larvae. All bioassays were carried out at constant temperatures of 10, 15, and 20 °C. Conidia of individual isolates demonstrated differences in germination after a 24-h long incubation at all evaluated temperatures. At 20 °C, the germination ranged from 67 to 100 % and at 15 °C from 5.33 to 46.67 %. At 10 °C, no germination was observed after 24 h; nevertheless, it was 8.67-44.67 % after 48 h. In terms of radial growth, the culture diameters and the associated production of spores of all isolates increased with increasing temperature. At 10 °C, sporulation was observed in three isolates while all remaining cultures appeared sterile. Three weeks post-inoculation, conidia of all assessed isolates caused 100 % cumulative mortality of treated larvae of T. molitor at 15 and 20 °C with the exception of isolate 110108 that induced 81.33 % mortality at 15 °C. At 10 °C, larval cumulative mortality ranged from 6.67 to 85.33 % depending on the isolate. Isolates 110108 and 110111 showed significantly slower outset and a much lower rate of infection at all temperatures compared to other tested isolates of M. anisopliae. The bioassays were carried out with the purpose to sort and select indigenous isolates of M. anisopliae useful as biocontrol agents in their original habitat.

  7. Metarhizium anisopliae var. Anisopliae Pr1A基因的克隆表达及抗血清研究%Cloning and Expressing Pr1A Gene of the Entomopathogene fungus Metarhizium anisopliae var. anisopliae and Researching Its Antiserum

    Institute of Scientific and Technical Information of China (English)

    宋妍; 张世清; 黄俊生

    2006-01-01

    采用RT-PCR方法从本实验室分离筛选到的金龟子绿僵小孢变种Metarhizium anisopliae var.anisopliae中,扩增得到Pr1A基因全长,此基因全长为1242bp,经Blastn分析此基因序列与M.anisopliae的Pr1A基因(M73795)同源率为98%.以pET-22b(+)为基础载体,构建pET-Pr1A重组表达载体,在大肠杆菌(Escherichia.coli)BL 21(DE3)中进行表达.经SDS-PAGE分析,获得了约42kDa大小的重组目的蛋白,目的蛋白占表达总蛋白含量的63.2%.将表达的Pr1A蛋白切胶回收后制备成抗原,免疫家兔4次后,采血收集抗血清,用ELISA测定效价为1/10 000.结果表明,获得的抗体可用于更进一步的研究,将有利于我们进一步了解M.anisopliaeis的侵染机理,弄清楚各Pr蛋白酶的作用方式和对寄主的选择优势,提高生防控制的有效性.

  8. Interactions between isolates of the entomopathogenic fungus Metarhizium anisopliae and the entomopathogenic nematode Heterorhabditis bacteriophora JPM4 during infection of the sugar cane borer Diatraea saccharalis (Lepidoptera: Pyralidae).

    Science.gov (United States)

    Acevedo, Juan Pablo Molina; Samuels, Richard Ian; Machado, Inês Ribeiro; Dolinski, Claudia

    2007-10-01

    Interactions between the nematode Heterorhabditis bacteriophora isolate JPM4 and the fungus Metarhizium anisopliae, isolates LPP45 and LPP39, were studied during dual infections of Diatraea saccharalis. Mortality, production of infective juveniles (IJs) and production of conidia were evaluated. A positive effect was demonstrated for host mortality in duel infections of JPM4 and LPP39, causing 100% mortality with LT(50) and LT(95) values of 1.8 and 2.8 days, respectively. Higher values were seen when using the nematode or fungi individually. However, a combination of JPM4+LPP39 caused a significant reduction in IJ production. The results show that faster time to death, a moderately virulent fungal isolate could be combined with the nematode, however at the expense of IJ production.

  9. Seletividade de Lecanicillium lecanii e Metarhizium anisopliae para larvas de primeiro ínstar de Ceraeochrysa cincta (Neuroptera: Chrysopidae em laboratório = Selectivicty of Lecanicillium lecanii and Metarhizium anisopliae for Ceraeochrysa cincta first instar larvae (Neuroptera: Chrysopidae in laboratory

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    Eliane Ribeiro Cardoso

    2007-10-01

    Full Text Available O objetivo deste trabalho foi avaliar a seletividade dos fungos Lecanicillium lecanii e Metarhizium anisopliae para larvas de Ceraeochrysa cincta, utilizando-se a aplicação por imersão e aaplicação em bases de vidro, formando filme seco (IOBC modificado. Nos dois métodos, foram aplicados os tratamentos: 1 - L. lecanii; 2 - M. anisopliae - 2.1x107 conídios viáveis mL-1; 3 - solução de Tween 80® a 0,05% como testemunha. Avaliou-se a mortalidade, duração médiado período larval, fecundidade e a viabilidade dos ovos. Todos os insetos mortos foram submetidos à assepsia, para confirmação da mortalidade causada pelo fungo. Em ambos os bioensaios, as mortalidades confirmadas pelos fungos foram iguais a zero. No filme seco o efeito total dos fungos sobre o predador foi classificado como pouco nocivo de acordo com as categorias propostas pelo método IOBC. L. lecanii e M. anisopliae não afetaram a viabilidade dos ovos de C. cincta, quando aplicados por imersão. Na concentração que foi estabelecida para a realização dos bioensaios, os fungos entomopatogênicos L. lecanii e M. anisopliae foram seletivos para larvas de 1o ínstar de C. cincta.This paper aimed to evaluate the selectivity of the fungi Lecanicillium lecanii and Metarhizium anisopliae for Ceraeochrysa cincta. Two bioassays were developed: 1st application of dry filmproposed by the International Organization for Biological Control IOBC (modified - they were applied in glass bases, forming dry film. 2nd immersion application. In the two methods, the following treatments were applied: 1- watery suspension of L. lecanii; 2- the same for M. anisopliae (both at 2.1x107 viable conidia mL-1; and 3- Tween 80® 0.05% watery solution. The larvae mortality, the average duration of the larvae period, the females fecundity and the eggsviability were evaluated. All the dead insects were submitted to asepsis to confirm the mortality caused by the fungi. In both bioassays, the mortality

  10. Distribuição volumétrica de calda contendo Metarhizium anisopliae Volumetric distribution of spray containing Metarhizium anisopliae

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    Haroldo Xavier Linhares Volpe

    2012-11-01

    Full Text Available O controle microbiano de Mahanarva fimbriolata é de suma importância para o manejo dessa praga em cana-de-açúcar, porém carece de melhorias quanto à tecnologia de aplicação. O trabalho teve como objetivo definir um modelo de bico de pulverização adequado para o controle de M. fimbriolata, com base no padrão de distribuição de calda aspergida e espaçamento entre bicos. Utilizaram-se os bicos TF4, TTI e AIUB11004VS e caldas a base de fungo Metarhizium anisopliae sem adjuvante e com Agral a 2% e 4%. Avaliou-se o espaçamento entre bicos baseado na construção de curvas de deposição, considerando CV máximo de 10%; ângulo de abertura e vazão dos bicos. Os maiores espaçamentos foram de 85cm em calda com 2% de adjuvante para o bico TF4, 70cm para AIUB11004VS sem adição de adjuvante e 55cm para o bico TTI, independente da calda. Em relação ao ângulo de abertura, houve apenas diferença entre os bicos testados em uma mesma calda. A vazão foi maior para a calda com 4% de adjuvante para os bicos TF4 e TTI, sendo que AIUB11004VS apresentou menores vazões em relação aos outros modelos para calda com 2% e 4% de adjuvante. Conclui-se que o modelo AIUB11004VS é uma importante ferramenta operacional, visando o controle de M. fimbriolata, por apresentar menor consumo de calda e bons resultados de distância entre bicos.The microbial control of Mahanarva fimbriolata is very important to management of this pest in sugarcane crops, however lacks improvement in spraying technology. The work aimed to evaluate a nozzle model adequate to control M. fimbriolata, based on pattern of spray distribution and the space between nozzles. TF4, TTI and AIUB11004VS nozzles containing the fungus Metarhizium anisopliae without adjuvant, with Agral 2% and 4% were used in the assay. We evaluated spacing between nozzles by constructing deposition curves considering maximum CV of 10%, spray angle and nozzle flow for different sprays. The larger nozzle

  11. Effect of two dosages of Metarhizium anisopliae var. acridum against Rhammatocerus schistocercoides Rehn Efeito de duas dosagens de Metarhizium anisopliae var. acridum contra Rhammatocerus schistocercoides Rehn

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    Marcos Rodrigues de Faria

    2002-11-01

    Full Text Available The fungus Metarhizium anisopliae var. acridum, strain CG 423, was tested under field conditions against the gregarious grasshopper Rhammatocerus schistocercoides (Rehn (Orthoptera: Acrididae. Conidia formulated in a racemic mixture of soybean oil and kerosene were sprayed under field conditions using an ultralow-volume hand-held atomizer Ulva Plus adjusted to deliver 2.9 L/ha. Bands composed of 2nd instar nymphs were treated with either 5.0x10(12 or 1.0x10(13 viable conidia/ha. The number of insects in each band was estimated at day one following spraying and by the end of the field trial (15 to 16 days post-treatment. Reductions in population size reached, in average, 65.8% and 80.4% for bands treated with the higher and lower dosage, respectively. For both dosages, total mortality rates of insects collected at two days post-application, and kept in cages for 14 days under lab conditions, showed no significant differences as compared to that obtained with insects collected immediately after spraying. Healthy insects were fed to native grasses sprayed on the field with 1.0x10(13 viable conidia/ha. Mortality levels of the nymphs fed on grasses collected two and four days post-application were not affected when compared to nymphs fed on grasses collected immediately following application.O fungo Metarhizium anisopliae var. acridum, isolado CG 423, foi avaliado em condições de campo como agente de controle biológico do gafanhoto gregário Rhammatocerus schistocercoides (Rehn (Orthoptera: Acrididae. Conídios formulados em uma mistura racêmica de óleo de soja e querosene foram pulverizados no campo com a utilização de um atomizador rotativo manual Ulva Plus, calibrado para aplicação de 2,9 L/ha. Bandos com ninfas de 2º estádio foram tratados com o equivalente a 5,0x10(12 ou 1,0x10(13 conídios viáveis/ha. O número de insetos em cada bando foi estimado no 1º dia após a aplicação e ao final do experimento (15 a 16 dias ap

  12. Metarhizium anisopliae host-pathogen interaction: differential immunoproteomics reveals proteins involved in the infection process of arthropods.

    Science.gov (United States)

    Santi, Lucélia; Silva, Walter O B; Pinto, Antônio F M; Schrank, Augusto; Vainstein, Marilene H

    2010-04-01

    Metarhizium anisopliae is an entomopathogenic fungus well characterized for the biocontrol of a wide range of plagues. Its pathogenicity depends on the secretion of hydrolytic enzymes that degrade the host cuticle. To identify proteins involved in the infection process and in host specify, immunoproteomic analysis was performed using antiserum produced against crude extract of M. anisopliae cultured in the presence of Rhipicephalus (Boophilus) microplus and Dysdercus peruvianus cuticles. Spots detected using antisera produced against M. anisopliae cultured in cuticles and spore surface proteins, but not with antiserum against M. anisopliae cultured in glucose, were identified so as to give insights about the infection process. An MS/MS allowed the identification of proteases, like elastase, trypsin, chymotrypsin, carboxypeptidase and subtilisin (Pr1A, Pr1I and PR1J), chitinases, DNase I and proline-rich protein. Chymotrypsin and Pr1I were inferred as host specific, being recognized in D. peruvianus infection only. This research represents an important contribution to the understanding the adaptation mechanisms of M. anisopliae to different hosts.

  13. Potential capacity of Beauveria bassiana and Metarhizium anisopliae in the biosorption of Cd2+ and Pb2+.

    Science.gov (United States)

    Hussein, Khalid A; Hassan, Sedky H A; Joo, Jin Ho

    2011-01-01

    In this study Beauveria bassiana and Metarhizium anisopliae were used as inexpensive and efficient biosorbents for Pb(II) and Cd(II) from aqueous metal solutions. The effects of various physicochemical factors on Pb(II) and Cd(II) biosorption by B. bassiana and M. anisopliae were studied. The optimum pH for Cd(II) and Pb(II) biosorption by two fungal species was achieved at pH 6.0 for Pb(II) and 5.0 Cd(II) at a constant time of 30 min. The nature of fungal biomass and metal ion interactions was evaluated by Fourier transform infrared. The maximum adsorption capacities (q(max)) calculated from Langmuir isotherms for Pb(II), and Cd(II) uptake by B. bassiana were 83.33±0.85, and 46.27±0.12 mg/g, respectively. However, the q(max) obtained for Pb(II) uptake by M. anisopliae was 66.66±0.28 mg/g, and 44.22±0.13 mg/g for Cd(II). B. bassiana showed higher adsorption capacity compared to M. anisopliae. The data obtained imply the potential role of B. bassiana and M. anisopliae for heavy metal removal from aqueous solutions.

  14. Temperature requirements of four entomopathogenic fungi

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    Ryszard Miętkiewski

    2014-08-01

    Full Text Available Infection potential of Beauvaria bassiana, Metarhizium anisopliae, Paecilomyces farinosus and P. fumosoroseus to G. mellonella larvae in pine litter was established. The growth of these fungi on the Czapek's Dox medium exposed to different temperature was determined.

  15. Neem oil increases the efficiency of the entomopathogenic fungus Metarhizium anisopliae for the control of Aedes aegypti (Diptera: Culicidae) larvae.

    Science.gov (United States)

    Gomes, Simone A; Paula, Adriano R; Ribeiro, Anderson; Moraes, Catia O P; Santos, Jonathan W A B; Silva, Carlos P; Samuels, Richard I

    2015-12-30

    Entomopathogenic fungi are potential candidates for use in integrated vector management and many isolates are compatible with synthetic and natural insecticides. Neem oil was tested separately and in combination with the entomopathogenic fungus Metarhizium anisopliae against larvae of the dengue vector Aedes aegypti. Our aim was to increase the effectiveness of the fungus for the control of larval mosquito populations. Commercially available neem oil was used at concentrations ranging from 0.0001 to 1%. Larval survival rates were monitored over a 7 day period following exposure to neem. The virulence of the fungus M. anisopliae was confirmed using five conidial concentrations (1 × 10(5) to 1 × 10(9) conidia mL(-1)) and survival monitored over 7 days. Two concentrations of fungal conidia were then tested together with neem (0.001%). Survival curve comparisons were carried out using the Log-rank test and end-point survival rates were compared using one-way ANOVA. 1% neem was toxic to A. aegypti larvae reducing survival to 18% with S50 of 2 days. Neem had no effect on conidial germination or fungal vegetative growth in vitro. Larval survival rates were reduced to 24% (S50 = 3 days) when using 1 × 10(9) conidia mL(-1). Using 1 × 10(8) conidia mL(-1), 30% survival (S50 = 3 days) was observed. We tested a "sub-lethal" neem concentration (0.001%) together with these concentrations of conidia. For combinations of neem + fungus, the survival rates were significantly lower than the survival rates seen for fungus alone or for neem alone. Using a combination of 1 × 10(7) conidia mL(-1) + neem (0.001%), the survival rates were 36%, whereas exposure to the fungus alone resulted in 74% survival and exposure to neem alone resulted in 78% survival. When using 1 × 10(8) conidia mL(-1), the survival curves were modified, with a combination of the fungus + neem resulting in 12% survival, whilst the fungus alone at this concentration also

  16. Laboratory Bioassay of Iranian Isolates of Entomopathogenic Fungus Metarhizium anisopliae (Metsch. Sorokin (Ascomycota: Hypocreales against two Species of Storage Pest

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    Adel Khashaveh

    2013-03-01

    Full Text Available M-B 12.00 Normal 0 false false false EN-US X-NONE FA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Times New Roman","serif";} The susceptibility of adults of Tribolium castaneum Herbst (Coleoptera: Tenebrionidae and Oryzaephilus surinamensis L. (Coleoptera: Silvanidae to three Iranian Isolates of the entomopathogenic fungi Metarhizium anisopliae (Metsch. Sorokin was evaluated through bioassays with direct immersion under laboratory conditions. For each isolates, five aqueous suspensions were prepared in a logarithmic series in Tween 80 (0.05% v/v. Results showed that adult of both species were susceptible to all isolates of M. anisopliae. For all three isolates, mortality percentage of the two species increased with increasing conidial concentration and significant difference was observed between concentrations. The corrected cumulative mortality of adult of O. surinamensis 10 days after immersion ranged from 12.38 to 85.84%, 18.6 to 62.83% and 10.63 to 77.87% for different concentrations of DEMI001, IRAN 715C and IRAN 1018C, respectively. These amounts for T. castaneum varied from 31.07 to 74.78%, 26.02 to 75.61% and 23.33 to 89.99% for different concentrations of DEMI001, IRAN 715C and IRAN 1018C, respectively. The parameters of probit analysis demonstrated non-overlap of 95% confidence limits of LC50 and LC95 and significant difference was observed among three isolates tested against each insect. The lowest and the highest LC50 and LC95 values were observed in the isolates DEMI001 for O. surinamensis (3/1×105 and 1/5×108 and IRAN 715C for T. castaneum (6.2×108 and 6.9×1014, respectively. This

  17. The combination of the entomopathogenic fungus Metarhizium anisopliae with the insecticide Imidacloprid increases virulence against the dengue vector Aedes aegypti (Diptera: Culicidae).

    Science.gov (United States)

    Paula, Adriano R; Carolino, Aline T; Paula, Cátia O; Samuels, Richard I

    2011-01-25

    Dengue fever transmitted by the mosquito Aedes aegypti, is one of the most rapidly spreading insect borne diseases, stimulating the search for alternatives to current control measures. The dengue vector A. aegypti has received less attention than anophelene species, although more than 2.5 billion people are at risk of infection worldwide. Entomopathogenic fungi are emerging as potential candidates for the control of mosquitoes. Here we continue our studies on the pathogenicity of the entomopathogenic fungus Metarhizium anisopliae against adult A. aegypti females. With the aim of further reducing mean survival times of A. aegypti exposed to fungus impregnated surfaces, a sub-lethal concentration of the neonicotinoid insecticide Imidacloprid (IMI) was added to fungal suspensions. A sub-lethal concentration of IMI that did not significantly alter the daily survival rates or mean survival percentages of mosquitoes was identified to be 0.1 ppm. This sub-lethal concentration was combined with M. anisopliae conidia (1 × 10(9) conidia mL(-1)). Both the combined treatment and the conidia alone were able to reduce the survival of A. aegypti compared with untreated or IMI treated mosquitoes. Importantly, mosquito survival following exposure to the combined treatment for 6 and 12 hrs was significantly reduced when compared with mosquitoes exposed to conidia alone. This is the first time that a combination of an insecticide and an entomopathogenic fungus has been tested against A. aegypti. Firstly, the study showed the potential of IMI as an alternative to the currently employed pyrethroid adulticides. Secondly, as an alternative to applications of high concentrations of chemical insecticides, we suggest that adult A. aegypti could be controlled by surface application of entomopathogenic fungi and that the efficiency of these fungi could be increased by combining the fungi with ultra-low concentrations of insecticides, resulting in higher mortality following relatively short

  18. The combination of the entomopathogenic fungus Metarhizium anisopliae with the insecticide Imidacloprid increases virulence against the dengue vector Aedes aegypti (Diptera: Culicidae

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    Samuels Richard I

    2011-01-01

    Full Text Available Abstract Background Dengue fever transmitted by the mosquito Aedes aegypti, is one of the most rapidly spreading insect borne diseases, stimulating the search for alternatives to current control measures. The dengue vector A. aegypti has received less attention than anophelene species, although more than 2.5 billion people are at risk of infection worldwide. Entomopathogenic fungi are emerging as potential candidates for the control of mosquitoes. Here we continue our studies on the pathogenicity of the entomopathogenic fungus Metarhizium anisopliae against adult A. aegypti females. With the aim of further reducing mean survival times of A. aegypti exposed to fungus impregnated surfaces, a sub-lethal concentration of the neonicotinoid insecticide Imidacloprid (IMI was added to fungal suspensions. Results A sub-lethal concentration of IMI that did not significantly alter the daily survival rates or mean survival percentages of mosquitoes was identified to be 0.1 ppm. This sub-lethal concentration was combined with M. anisopliae conidia (1 × 109 conidia mL-1. Both the combined treatment and the conidia alone were able to reduce the survival of A. aegypti compared with untreated or IMI treated mosquitoes. Importantly, mosquito survival following exposure to the combined treatment for 6 and 12 hrs was significantly reduced when compared with mosquitoes exposed to conidia alone. Conclusions This is the first time that a combination of an insecticide and an entomopathogenic fungus has been tested against A. aegypti. Firstly, the study showed the potential of IMI as an alternative to the currently employed pyrethroid adulticides. Secondly, as an alternative to applications of high concentrations of chemical insecticides, we suggest that adult A. aegypti could be controlled by surface application of entomopathogenic fungi and that the efficiency of these fungi could be increased by combining the fungi with ultra-low concentrations of insecticides

  19. African water storage pots for the delivery of the entomopathogenic fungus Metarhizium anisopliae to the malaria vectors Anopheles gambiae s.s. and Anopheles funestus.

    Science.gov (United States)

    Farenhorst, Marit; Farina, Daniela; Scholte, Ernst-Jan; Takken, Willem; Hunt, Richard H; Coetzee, Maureen; Knols, Bart G J

    2008-06-01

    We studied the use of African water storage pots for point source application of Metarhizium anisopliae against the malaria vectors Anopheles gambiae s.s. and An. funestus. Clay pots were shown to be attractive resting sites for male and female An. gambiae s.s. and were not repellent after impregnation with fungus. M. anisopliae was highly infective and virulent after spray application inside pots. At a dosage of 4 x 10(10) conidia/m(2), an average of 95 +/- 1.2% of An. gambiae s.s. obtained a fungal infection. A lower dosage of 1 x 10(10) conidia/m(2) infected an average of 91.5 +/- 0.6% of An. gambiae s.s. and 91.8 +/- 1.2% of An. funestus mosquitoes. Fungal infection significantly reduced mosquito longevity, as shown by differences between survival curves and LT(50) values. These pots are suitable for application of entomopathogenic fungi against malaria vectors and their potential for sustainable field implementation is discussed.

  20. Trehalose and trehalose-hydrolyzing enzyme in the haemolymph of Locusta migratoria infected with Metarhizium anisopliae strain CQMa102

    Institute of Scientific and Technical Information of China (English)

    HUA ZHAO; ZHONG-KANG WANG; YOU-PING YIN; YAN-LING LI; ZHEN-LUN LI; GUO-XIONG PENG; YU-XIAN XIA

    2007-01-01

    Topical application of the Metarhizium anisopliae var. acridum specialist strain CQMa102 to the locust Locusta migratoria man ilensis results in changes of the concentrations of trehalose and glucose in the haemolymph. Micrographs of the locust haemolymph shows Metarhizium anisopliae can effectivly penetrate the external skeleton of locust and after 2 days infection, the hyphae body will appear in the haemolymph of infected insects. The time in decrease of trehalose concentration coincided with that in increase of trehalose-hydrolysing enzyme activity in the haemolymph of the fungus-infected insects. Overlay gel analysis indicated there was considerably more trehalose-hydrolysing activity in the haemolymph of locusts infected by fungus than in controls. A comparable isoform was identified in in vitro culture of the fungus, suggesting a fungal origin for the in vivo enzyme. Haemolymph trehalose decreased significantly during mycosis of locusts by M. anisopliae. All these results suggested that this fungus may take advantage of competing nutrient utilization against the insect by its trehalose-hydrolyzing enzyme secretion. It may provide fundamental knowledge for fungal pathogenesis.

  1. POTENCIAL DE BLASTOSPOROS DE Metarhizium anisopliae PARA O CONTROLE DE ADULTOS DE Aedes aegypti

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    Aline Teixeira Carolino

    2017-04-01

    Full Text Available Os conídios do fungo entomopatogênico Metarhizium anisopliae são capazes de infectar desde o ovo até a fase adulta do mosquito Aedes aegypti. Em cultura líquida, fungos entomopatogênicos são capazes de produzir blastosporos, um propágulo com características hidrofílicas e às vezes mais virulento que conídios. No presente estudo foi observado pela primeira vez, que blastosporos secos quando formulados em óleo são eficientes para controle do mosquito adulto. Foi observada somente 1% de sobrevivência dos mosquitos tratados com blastosporos e 94% de sobrevivência do grupo controle. Estudos já evidenciaram que formulações em óleo podem melhorar a eficiência de fungos entomopatogênicos durante o processo de infecção, facilitando a adesão do mesmo com o tegumento dos insetos. Pelo fato dos blastosporos serem hidrofílicos, a película formada pelo óleo ao redor do fungo, favorece a interação entre o fungo e o inseto, levando à infecção dos adultos.

  2. Field bioassay of Metarhizium anisopliae strains to control the poultry red mite Dermanyssus gallinae.

    Science.gov (United States)

    Tavassoli, M; Allymehr, M; Pourseyed, S H; Ownag, A; Bernousi, I; Mardani, K; Ghorbanzadegan, M; Shokrpoor, S

    2011-06-10

    The poultry red mite, Dermanyssus gallinae is one of the most economically deleterious ectoparasite of laying hens worldwide. To evaluate the efficacy of three strains (V245, 3247 and 715C) of entomopathogenic fungus Metarhizium anisopliae with potential as acaricides against D. gallinae, this investigation was carried out in a commercial caged laying poultry farm in Naghedeh, West Azarbaijan of Iran. The parasite infestation already existed in the farm. Sunflower oil suspension of all fungal strains, each in two concentrations (1×10(7) and 1×10(9) conidia/ml) were used separately as spray on hens and cages, and in the control group the cages were only sprayed with sunflower oil and sterile distilled water. For estimating the population rate of mites before and after treatment, special cardboard traps were fixed to cages during a 1-month period. The traps were placed on weeks -1, 0, 1, 2 and 3 and always removed after 1 w. The results showed that the population rates post fungal treatment with the lower concentration were not significantly different compared to the control group. However, the reduction in mite numbers induced by all three strains at the concentration of 1×10(9) conidia/ml was significantly higher than the control (Pgallinae.

  3. Enhancing Growth of Vigna radiata in the Presence of Pseudomonas aeruginosa Biopolymer and Metarhizium anisopliae Spores

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    Bhagwan N. Rekadwad

    2016-01-01

    Full Text Available Exopolysaccharide producing Pseudomonas aeruginosa NCIM 2945 (PANCL belonging to gamma-proteobacterium and entomopathogenic fungus Metarhizium anisopliae MCC 1129 (MAMCC belonging to Ascomycota were studied for their morphological features biochemical characteristics and plant growth promotion ability. Optimum growth of PANCL was recorded after 24 h at temperature 30°C and pH 7.0. Gram-negative PANCL appeared as white in color, one mm size, circular, opaque, and nonconsistent elevated colonies with entire margin. It has utilized dextrose, fructose, maltose, and sorbitol as carbon source and produced acid in the medium. PANCL was sensitive to Polymyxin B (300 µgm/disc followed by Neomycin (30 µgm/disc, Gentamycin (10 µgm/disc, and Chloramphenicol (30 µgm/disc. PANCL has secreted extracellular lipase, amylase, protease, and exopolysaccharides (EPS. Another fungal strain MAMCC sporulated after 168 h at temperature 30°C and pH 7.0. MAMCC has septate-white mycelium and bears dirty green colored spores. Growth of MAMCC was enhanced in the presence of Neem and Karela-Amla oil (0.1 mL each. Extracellular polysaccharide produced by PANCL and spores of MAMCC promoted growth of dicotyledon Vigna radiata (Mung individually as well as in consortium. Considerable increase in dry weight of Vigna radiata was recorded. Thus, reported PANCL and MAMCC strains have promoted growth Vigna radiata and may be a solution for sustainable agriculture.

  4. Expression of a chitinase gene from Metarhizium anisopliae in tobacco plants confers resistance against Rhizoctonia solani.

    Science.gov (United States)

    Kern, Marcelo Fernando; Maraschin, Simone de Faria; Vom Endt, Débora; Schrank, Augusto; Vainstein, Marilene Henning; Pasquali, Giancarlo

    2010-04-01

    The chit1 gene from the entomopathogenic fungus Metarhizium anisopliae, encoding the endochitinase CHIT42, was placed under the control of the CaMV 35S promoter, and the resulting construct was transferred to tobacco. Seventeen kanamycin-resistant transgenic lines were recovered, and the presence of the transgene was confirmed by polymerase chain reactions and Southern blot hybridization. The number of chit1 copies was determined to be varying from one to four. Copy number had observable effects neither on plant growth nor development. Substantial heterogeneity concerning production of the recombinant chitinase, and both general and specific chitinolytic activities were detected in leaf extracts from primary transformants. The highest chitinase activities were found in plants harboring two copies of chit1 inserts at different loci. Progeny derived from self-pollination of the primary transgenics revealed a stable inheritance pattern, with transgene segregation following a mendelian dihybrid ratio. Two selected plants expressing high levels of CHIT42 were consistently resistant to the soilborne pathogen Rhizoctonia solani, suggesting a direct relationship between enzyme activity and reduction of foliar area affected by fungal lesions. To date, this is the first report of resistance to fungal attack in plants mediated by a recombinant chitinase from an entomopathogenic and acaricide fungus.

  5. Pathogenicity of Metarhizium anisopliae for Ceratitis capitata (Wied.) (Diptera: Tephritidae) in soil with different pesticides.

    Science.gov (United States)

    Mochi, Dinalva A; Monteiro, Antonio C; De Bortoli, Sergio A; Dória, Háyda O S; Barbosa, José C

    2006-01-01

    This research intended to investigate if the presence of pesticides in the soil could affect the pathogenicity of Metarhizium anisopliae Metsch. (Sorokin) for Ceratitis capitata (Wied.) and assess the effect of conidia application as suspension or dry conidia. The fungicides chlorothalonyl and tebuconazol, the acaricide abamectin, the insecticide trichlorfon, and the herbicide ametrin were applied at the manufacturer-recommended doses. Soil samples were placed in glass flasks and were given the fungus as conidial suspension or dry. After pesticide application, 20 3rd-instar larvae were placed in the soil. The flasks were sealed with voile fabric and incubated at 27 +/- 0.5 masculineC for nine days, until adult emergence; incubation continued for four more days at room temperature. The total insect survival was significantly affected and pathogenic activity was detected from the pupa stage on. Pupa survival was reduced (Ptebuconazole reduced (86.2% and 82.5%, respectively) the survival period of C. capitata compared to the control (95.0%). The techniques used for conidia application did not influence the total insect survival rate, but conidial suspension applied on soil surface reduced survival during the pupae and adult phases.

  6. Hemocyte characterization of Nasutitermes coxipoensis (Holmgren) (Isoptera: Termitidae) workers and hemocyte evaluation after parasitism by Metarhizium anisopliae; Caracterizacao dos hemocitos de operarios de Nasutitermes coxipoensis (Holmgren) (Isoptera: Termitidae) e avaliacao hemocitaria apos parasitismo por Metarhizium anisopliae

    Energy Technology Data Exchange (ETDEWEB)

    Cunha, Franklin M.; Wanderley-Teixeira, Valeria; Albuquerque, Auristela C. [Universidade Federal Rural de Pernambuco (UFRPE), Recife, PE (Brazil). Programa de Pos-Graduacao em Entomologia Agricola], e-mail: ukento@yahoo.com.br; Teixeira, Alvaro A.C. [Universidade Federal Rural de Pernambuco (UFRPE), Recife, PE (Brazil). Dept. de Morfologia e Fisiologia Animal], e-mail: valeria@dmfa.ufrpe.br, e-mail: auritermes@yahoo.com.br; Alves, Luiz C. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Lab. de Imunopatologia Keizo Asami (LIKA); Lima, Elza A.L.A. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de Micologia. Lab. de Controle Biologico

    2009-03-15

    We aimed to characterize the morphology and ultrastructure of hemocytes of Nasutitermes coxipoensis (Holmgren) workers and to quantify the cell types 24h, 48h and 72h after inoculation with Metarhizium anisopliae. Six hemocytes types were identified, plasmatocyte, granulocyte, spherulocyte, prohemocyte, adipohemocyte and eonocytoid Hemocytes did not present any morphological alteration at the several observation periods, but they did have a change in their abundance, as observed for spherulocytes, adipohemocytes and eonocytoids at all intervals, and for plasmatocytes and granulocytes at 48h after host inoculation. We argue on the possible reasons and implications of the observed changes. (author)

  7. First report of Metarhizium anisopliae IP 46 pathogenicity in adult Anopheles gambiae s.s. and An. arabiensis (Diptera; Culicidae

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    Lyimo Issa N

    2009-12-01

    Full Text Available Abstract The entomopathogenic fungus Metarhizium anisopliae isolate IP 46, originating from a soil sample collected in 2001 in the Cerrado of Central Brazil, was tested for its ability to reduce the survival of adult male and female Anopheles gambiae s.s. and An. arabiensis mosquitoes. A 6-h exposure to the fungus coated on test paper at a concentration of 3.3 × 106 conidia cm-2 reduced the daily survival of both mosquito species (HR = 3.14, p An. gambiae s.s relative to An. arabiensis (HR = 1.38, p 95% of mosquito cadavers in the treatment groups. The results indicate that M. anisopliae IP 46 has the potential to be a bio-control agent for African malaria vector species, and is a suitable candidate for further research and development.

  8. Comparative analysis of the Metarhizium anisopliae secretome in response to exposure to the greyback cane grub and grub cuticles.

    Science.gov (United States)

    Manalil, Nirupama Shoby; Junior Téo, V S; Braithwaite, K; Brumbley, S; Samson, P; Helena Nevalainen, K M

    2010-08-01

    Metarhizium anisopliae is a well-characterized biocontrol agent of a wide range of insects including cane grubs. In this study, a two-dimensional (2D) electrophoresis was used to display secreted proteins of M. anisopliae strain FI-1045 growing on the whole greyback cane grubs and their isolated cuticles. Hydrolytic enzymes secreted by M. anisopliae play a key role in insect cuticle-degradation and initiation of the infection process. We have identified all the 101 protein spots displayed by cross-species identification (CSI) from the fungal kingdom. Among the identified proteins were 64-kDa serine carboxypeptidase, 1,3 beta-exoglucanase, Dynamin GTPase, THZ kinase, calcineurin like phosphoesterase, and phosphatidylinositol kinase secreted by M. ansiopliae (FI-1045) in response to exposure to the greyback cane grubs and their isolated cuticles. These proteins have not been previously identified from the culture supernatant of M. anisopliae during infection. To our knowledge, this the first proteomic map established to study the extracellular proteins secreted by M. ansiopliae (FI-1045) during infection of greyback cane grubs and its cuticles.

  9. Survival of anopheline eggs and their susceptibility to infection with Metarhizium anisopliae and Beauveria bassiana under laboratory conditions.

    Science.gov (United States)

    Luz, Christian; Mnyone, Ladslaus L; Russell, Tanya L

    2011-09-01

    The viability of Anopheles gambiae sensu stricto and Anopheles arabiensis (Diptera: Culicidae) eggs over time and the ovicidal activity of Beauveria bassiana (Ascomycota: Cordycipitaceae) and Metarhizium anisopliae (Ascomycota: Clavicipitaceae) were investigated. Eggs were incubated in soil or leaf litter for up to 12 weeks at 26°C and 75%, 86% or >98% relative humidity (RH). Eggs were treated topically with M. anisopliae ICIPE-30 or B. bassiana I93-825 conidia in either water or oil-in-water formulations. Survival of eggs whether treated or not with fungus was similar, and untreated eggs generally did not survive longer than 2 weeks regardless of the substrate or humidity tested. After a minimal 5-day exposure, M. anisopliae at 5 × 10(6) conidia/cm(2) clearly reduced the number of larvae. The efficacy of the fungus increased when it was oil-in-water formulated, and eclosion was completely prevented regardless of the conidial concentration (10(5)-10(7) conidia/cm(2)) after a 10-day exposure in soils at >98% RH. Treatment of eggs with B. bassiana, however, failed to reduce the number of eclosing larvae. This is the first demonstration of the ovicidal activity by M. anisopliae against either A. gambiae s. s. or A. arabiensis and the results underline the potential of this fungus against anopheline mosquitoes.

  10. Virulence and mycotoxic effects of Metarhizium anisopliae on Mahogany shoot borer, Hypsipyla robusta (Lepidoptera: Pyralidae)

    Institute of Scientific and Technical Information of China (English)

    M.Balachander; O.K.Remadevi; T.O.Sasidharan; N.Sapna Bai

    2012-01-01

    Developing appropriate control measures for the Mahogany shoot borer,Hypsipyla robusta Moore has become increasingly important due to the severe damaging effect of the pest on the establishment of the saplings of Swietenia mahagoni Jacq (Sapindales:Meliaceae).Existing management methods are largely limited to silvicultural practices and spraying of chemical insecticides.To identify a potential fungal biocontrol agent,we compared the virulence of six native and two standard ARSEF isolates of Metarhizium anisopliae Metsch.against this pest.The average survival time and conidial yield of IWST-Ma7 was higher (6.2 to 7.3 days and 4.9 to 4.7 × 105 conidia/ml) than the standards.Significant difference in sporulation on the cadavers between isolates,doses and incubation periods were substantiated for the selection of potential strain.The mycotoxic effects of crude soluble protein extract when incorporated in the artificial diet,the ARSEF 2596 and ARSEF 3603showed LD50 value of 3.7% and 5.6%.However,IWST-Ma7 was highly lethal with significant lowest LD50 value of 2.6%.The enzyme activity of IWST-Ma7 was highest for chitinase,CDA,protease and lipase viz.,1.90 U/mg,1.80 U/mg,0.98 U/mg and 0.80 U/mg respectively.However the enzyme activity of chitinase and Chitin deacetylase assay for all the isolates was significantly higher than protease and lipase activity.The ITS regions (5.8S rDNA and 28S rDNA) of seven isolates of M.anisopliae were amplified using the ITS1 and ITS4 primers which was a unique fragment of approximately 550 bp.Based on ITS regions,phylogenetic tree have been constructed and the isolates have been grouped in to 5 clades.The virulence and mycotoxic effects of different isolates could rationally be used to employ them for the management of the mahogany borer.

  11. African water storage pots for the delivery of the Entomopathogenic fungus Metarhizium anisopliae to the Malaria vectors Anopheles gambiae s.s. and Anopheles funestus

    NARCIS (Netherlands)

    Farenhorst, M.; Farina, D.; Scholte, E.J.; Takken, W.; Hunt, R.H.; Coetzee, M.; Knols, B.G.J.

    2008-01-01

    We studied the use of African water storage pots for point source application of Metarhizium anisopliae against the malaria vectors Anopheles gambiae s.s. and An. funestus. Clay pots were shown to be attractive resting sites for male and female An. gambiae s.s. and were not repellent after impregnat

  12. Inhibition of the entomopathogenic fungus Metarhizium anisopliae in vitro by the bed bug defensive secretions (E)-2-hexenal and (E)-2-octenal

    Science.gov (United States)

    The two major aldehydes (E)-2-hexenal and (E)-2-octenal emitted as defensive secretions by bed bugs Cimex lectularius L. (Hemiptera: Cimicidae), inhibit the in vitro growth of Metarhizium anisopliae (Metsch.) Sokorin (Hypocreales: Clavicipitaceae). These chemicals inhibit fungal growth by direct con...

  13. African water storage pots for the delivery of the Entomopathogenic fungus Metarhizium anisopliae to the Malaria vectors Anopheles gambiae s.s. and Anopheles funestus

    NARCIS (Netherlands)

    Farenhorst, M.; Farina, D.; Scholte, E.J.; Takken, W.; Hunt, R.H.; Coetzee, M.; Knols, B.G.J.

    2008-01-01

    We studied the use of African water storage pots for point source application of Metarhizium anisopliae against the malaria vectors Anopheles gambiae s.s. and An. funestus. Clay pots were shown to be attractive resting sites for male and female An. gambiae s.s. and were not repellent after

  14. Réponse des stades larvaires de Helicoverpa armigera (Hübner (Lepidoptera : Noctuidae à l'application de champignons entomopathogènes Metarhizium anisopliae et Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    Tamò, M.

    2012-01-01

    Full Text Available Response of the nymphs of Helicoverpa armigera (Hübner (Lepidoptera: Noctuidae to entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana. Two experiments on dose/mortality response between the instars of Helicoverpa armigera and two strains of entomopathogenic fungi, Metarhizium anisopliae (Met 31 and Beauveria bassiana (Bb 11 were carried out in laboratory conditions. In the first experiment, M. anisopliae Met 31 was tested on the third instar of H. armigera, while in the second experiment, both Met 31 and Bb 11 were tested on the fourth instar. In all the experiments, the following different doses of conidia per insect were used: 104, 105, 106, 107. The following parameters were measured: mortality and sporulation rates, the number of pupae formed and the number of adults that emerged. Abbott's formula was used to correct the treatment mortality rates. LD50 was determined using Cox-regression. For the third instar in experiment one, no significant difference was observed between high doses (106 and 107 conidia per insect. For instar L4, only the dose of 107 conidia per insect showed high mortality rates (74%. For the strain Bb 11, in spite of the variation observed between the mortality rates induced by high doses (106 and 107 conidia per insect, no significant difference was recorded at the 5% level. No mycosis was observed from cadavers resulting from lower doses when tested on L4. The control recorded the highest numbers of pupae and adults. These two parameters were related to the level of dosage: the higher the dose, the lower the numbers of pupae and adults that emerged. For all the strains of fungi used, whatever the larval stage of H. armigera, the dose/mortality response was significant.

  15. Survival and mutant production induced by mutagenic agents in Metarhizium anisopliae Sobrevivência e obtenção de mutantes induzidos por agentes mutagênicos em Metarhizium anisopliae

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    V. Kava - Cordeiro

    1995-12-01

    Full Text Available A wild strain of Metarhizium anisopliae, an entomopathogenic fungus, was submitted to three mutagenic agents: gamma radiation, ultraviolet light and nitrous acid. Survival curves were obtained and mutants were selected using different mutagenic doses which gave 1 to 5% survival. Morphological and auxotrophic mutants were isolated. Morphological mutants were grouped in a class with yellow conidia and other with pale vinaceous conidia as opposed to the green wild type conidia. Auxotrophic mutants had requirements for vitamin and aminoacid biosynthesis. More than 58% of the total auxotrophk mutants required proline/aipnine. Gamma radiation showed to be the most efficient mutagenic agent giving 0.2% of auxotrophk mutants followed by ultraviolet light (0.12% and nitrous acid (0.06%.The conidial colour and auxotrophk mutants isolated until now from M. anisopliae were reviewed.Uma linhagem selvagem do fungo entomopatogênico Metarhizium anisopliae foi submetida à ação de três agentes mutagênicos: radiação gama, luz ultravioleta e ácido nitroso. Curvas de sobrevivência foram obtidas para cada mutagênicos utilizado e mutantes foram selecionados a partir de doses dos mutagênicos que proporcionassem de 1 a 5% de sobrevivência. Mutantes morfológicos para a coloração de conídios e mutantes auxotróficos foram isolados. Mutantes para coloração de conidios foram agrupados em duas classes, uma com conídios amarelos e outra com conídios vinho pálido. Os mutantes auxotróficos obtidos foram deficientes para aminoácidos e vitaminas e mais de 58% deles eram auxotróficos para prolina/argmina. Radiação gama foi o mutagênico mais eficiente com uma porcentagem de obtenção de mulantes auxotróficos de aproximadamente 0,2%, seguido pela luz ultravioleta (0.12% e pelo ácido nitroso (0.06%.Os mulantes morfológicos e auxotróficos obtidos até o momento em Metarhizium anisopliae foram revistos.

  16. In vitro effect of pesticides on the germination, vegetative growth, and conidial production of two strains of Metarhizium anisopliae.

    Science.gov (United States)

    Schumacher, Verona; Poehling, Hans-Michael

    2012-01-01

    Entomopathogenic fungi are widely used as biological control agents against a broad range of insect and arachnid pests. However, the control efficacy of entomopathogenic fungi is variable because of unfavourable and fluctuating environmental conditions and intrinsic factors. One strategy to enhance entomopathogenic fungi efficacy is a combined use of entomopathogenic fungi and low dosages of pesticides. These sub-lethal dosages of chemicals can increase the control efficiency of entomopathogenic fungi but only if they do not affect the fungi. Adverse effects could include the inhibition of germination and/or vegetative growth as well as conidiogenesis. The present study investigated the in vitro effects of different concentrations of fipronil, permethrin, imidacloprid, NeemAzal, and amitraz as potential candidates for combined applications on two strains of the entomopathogenic fungus Metarhizium anisopliae (MA). MA was inoculated on a medium amended with five different concentrations (0.32-200 ppm) of the abovementioned pesticides. The germination, vegetative growth, and sporulation were evaluated. The results showed, according to a physiology parameter compatibility classification, that all pesticides were compatible with both tested MA strains. Only fipronil in the higher dose rates of 40 and 200 ppm was close to moderately toxic to MA-7. Furthermore, only higher concentrations of the pesticides caused a slight inhibition (about 15%) of conidial germination and a reduction in colony size. Sporulation was reduced at most by approximately 50% by 40 or 200 ppm of fipronil or amitraz, respectively. Therefore, it is possible to use the tested pesticides in combination with either strain of MA for an integrated pest management approach. Studies on the effect of these combinations on target organisms are in progress.

  17. Commercial formulation of Metarhizium anisopliae for the control of Rhipicephalus microplus in a pen study.

    Science.gov (United States)

    Camargo, Mariana G; Marciano, Allan F; Sá, Fillipe A; Perinotto, Wendell M S; Quinelato, Simone; Gôlo, Patrícia S; Angelo, Isabele C; Prata, Márcia C A; Bittencourt, Vânia R E P

    2014-09-15

    The present study evaluated, for the first time, the effect of the commercial formulation Metarril(®) SP Organic of Metarhizium anisopliae plus 10% mineral oil to control Rhipicephalus microplus in a pen study. Three groups were formed with six animals each: the first group was exposed to Metarril(®) plus 10% mineral oil and 1% Tween 80; the second group was exposed to sterile distilled water, mineral oil and Tween 80 (oil control group); and the third group received no treatment (control group). The fungal formulation contained 1 × 10(8)conidiaml(-1). Each animal was sprayed with 3L of formulation. Fallen ticks were counted daily and a sample of 20 engorged females per day was incubated for assessment of biological parameters. Throughout the study period, Metarril(®) oil-based formulation showed an efficacy ranging from 19.20% to 67.39% in comparison with the control group; and from 8.18% to 61.38% in comparison with the oil control group. The average efficacy of Metarril(®) oil-based formulation was 47.74% and 40.89% in comparison with control and oil control groups, respectively. Changes in the biological parameters of engorged R. microplus females were observed in the first three days after treatment, with a significant reduction in hatching percentage and egg production index. We concluded that Metarril(®) SP Organic plus 10% mineral oil was efficient against R. microplus in pen studies. However, further in vivo studies are required to increase the efficacy and to establish a protocol for the use of this product in the field against the cattle tick.

  18. The production relationship of destruxins and blastospores of Metarhizium anisopliae with virulence againstPlutella xylostella

    Institute of Scientific and Technical Information of China (English)

    DONG Ting-yan; ZHANG Bo-wen; WENG Qun-fang; HU Qiong-bo

    2016-01-01

    Metarhizium anisopliaeas an essential entomopathogenic fungushas been known to produce destruxins (a kind of cyclo-peptidic mycotoxins) and blastospores in submerged culture. Blastospores and destruxins are candidates for in-secticides, but the relations of both productions and the impact factors are unclear yet. In this study, we investigated the effects of inoculums, rotation, dissolved oxygen (DO) on the productions of blastospores and destruxins A and B (DA and DB) in submerged culture ofM. anisopliaestrain MaQ01. The results indicated that DO levels were regulated by inoculum amounts and rotation speeds, meanwhile, the productions of DA, DB and blastospores were also closely inlfuenced by those factors. Totaly, when DO value was more than 40%, the higher productions of destruxins and blastospores were achieved, by contrast, lower than 40% of DO values resulted in lower productions. The regression analysis suggested that the productions of DA, DB and blastospores were positively correlated with the DO levels. Meanwhile, the positive correlations between the productions of DA or DB and blastospores were also found. Brielfy, when the rotation is 150 r min–1 and the inoculum is 1.0×106 spore mL–1, the DA, DB and blastospores achieved the best production of 61.81 mg mL–1, 24.74 mg mL–1 and 5.73×106 spore mL–1, respectively. In addition, the pathogenicities of blastospores and conidia against Plutela xylostela were bioassayed. The higher mortalities of P. xylostela were totaly recorded in blastospore treatments than in conidia treatments, especialy in lower dosages and earlier periods. Our research wil give some new insights to production of destruxins and blastospores by using M. anisopliae.

  19. A novel dodine-free selective medium based on the use of cetyl trimethyl ammonium bromide (CTAB) to isolate Beauveria bassiana, Metarhizium anisopliae sensu lato and Paecilomyces lilacinus from soil.

    Science.gov (United States)

    Posadas, Julieta B; Comerio, Ricardo M; Mini, Jorge I; Nussenbaum, Ana L; Lecuona, Roberto E

    2012-01-01

    This study evaluated the quaternary ammonium compound cetyl trimethyl ammonium bromide (CTAB) as an alternative to the chemically related dodecylguanidine (dodine) for the selective isolation of entomopathogenic fungi. Oatmeal agar (OA) with chloramphenicol was used as basal medium, and three concentrations of CTAB (0.5, 0.6, 0.7 g/L) were evaluated and compared against OA + 0.46 g/L dodine. Selective isolation and growth studies were performed with the entomopathogens Beauveria bassiana, Metarhizium anisopliae s.l. and Paecilomyces lilacinus and five common non-entomopathogenic non-target species. The three entomopathogenic fungi sporulated earlier on OA + 0.6 g/L CTAB than on OA + 0.46 g/L dodine, while none of the non-target fungi sporulated on OA + 0.6 g/L CTAB. All entomopathogenic fungal isolates grew on OA + 0.6 g/L CTAB, despite some intra-species variation, whereas non-target fungi showed no growth or sporulation. OA + 0.6 g/L CTAB resulted in an efficient medium to isolate B. bassiana, M. anisopliae s. l. and P. lilacinus from soil samples. Results of our study suggest that OA + 0.6 g/L CTAB is a suitable, simple and inexpensive to prepare medium to replace OA + 0.46 g/L dodine for the selective isolation of these fungi.

  20. Production and quality of conidia by Metarhizium anisopliae var. lepidiotum: critical oxygen level and period of mycelium competence.

    Science.gov (United States)

    Garcia-Ortiz, Nohemi; Tlecuitl-Beristain, Saúl; Favela-Torres, Ernesto; Loera, Octavio

    2015-03-01

    Mycoinsecticides application within Integral Pest Management requires high quantities of conidia, with the proper quality and resistance against environmental conditions. Metarhizium anisopliae var. lepidiotum conidia were produced in normal atmospheric conditions (21 % O2) and different concentrations of oxygen pulses (16, 26, 30, and 40 %); conidia obtained under hypoxic conditions showed significantly lower viability, hydrophobicity, and virulence against Tenebrio molitor larvae or mealworm, compared with those obtained under normal atmospheric conditions. Higher concentrations of oxygen (26 and 30 %) improved conidial production. However, when a 30 % oxygen concentration was applied, maximal conidial yields were obtained at earlier times (132 h) relative to 26 % oxygen pulses (156 h); additionally, with 30 % oxygen pulses, conidia thermotolerance was improved, maintaining viability, hydrophobicity, and virulence. Although conidial production was not affected when 40 % oxygen pulses were applied, viability and virulence were diminished in those conidia. In order to find a critical time for mycelia competence to respond to these oxidant conditions, oxygen pulses were first applied either at 36, 48, 60, and 72 h. A critical time of 60 h was determined to be the best time for the M. anisopliae var. lepidiotum mycelia to respond to oxygen pulses in order to increase conidial production and also to maintain the quality features. Therefore, oxygen-enriched (30 %) pulses starting at 60 h are recommended for a high production without the impairment of quality of M. anisopliae var. lepidiotum conidia.

  1. Secretome Analysis of Metarhizium anisopliae Under Submerged Conditions Using Bombyx mori Chrysalis to Induce Expression of Virulence-Related Proteins.

    Science.gov (United States)

    Rustiguel, Cynthia Barbosa; Rosa, José Cesar; Jorge, João Atílio; de Oliveira, Arthur Henrique Cavalcanti; Guimarães, Luis Henrique Souza

    2016-02-01

    The entomopathogenic fungus Metarhizium anisopliae is used to control insect pests. This species is specialized for the secretion of an enzymatic complex consisting of proteases, lipases, and chitinases related to pathogenicity and virulence. In this context, the secretomes of strains IBCB 167 and IBCB 384 of M. anisopliae var. anisopliae, grown under submerged fermentation in the presence of chrysalis as an inducer, were analyzed. Analysis of two-dimensional gels showed qualitative and quantitative differences between secreted proteins in both isolates. Around 102 protein spots were analyzed, and 76 % of the corresponding proteins identified by mass spectrometry were grouped into different classes (hydrolases, oxidases, reductases, isomerases, kinases, WSC domains, and hypothetical proteins). Thirty-three per cent of all the proteins analyzed were found to be common in both strains. Several virulence-related proteins were identified as proteases and mannosidases. Endo-N-acetyl-β-D-glucosaminidase expression was observed to be 10.14-fold higher for strain IBCB 384 than for strain IBCB 167, which may be an important contributor to the high virulence of IBCB 384 in Diatraea ssaccharalis. These results are important for elucidation of the host-pathogen relationship and the differences in virulence observed between the two strains.

  2. Selection of Beauveria bassiana (Bals. Vuill. and Metarhizium anisopliae (Metsch. Sorok. for the control of the mite Mononychellus tanajoa (Bondar

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    Barreto Rodrigo Soares

    2004-01-01

    Full Text Available The green mite, Mononychellus tanajoa (Bondar (Acari: Tetranychidae, is considered to be one of the key pests in cassava, Manihot esculenta Crants, leading to considerable field losses. In this study, ten Beauveria bassiana (Bals. Vuill. and ten Metarhizium anisopliae (Metsch. Sorok. isolates were evaluated with regard to their potential as biological control agents against adult M. tanajoa females. The total mortality percentage of M. tanajoa caused by B. bassiana ranged from 13.0 to 97.0%, with confirmed mortality rates extending from 9.0 to 91.0% and LT50 varying from 4.2 to 17.0 days. The M. anisopliae isolates showed total mortality percentages ranging from 12.0 to 45.0% with confirmed mortality rates extending from 8.0 to 45.0%, and LT50 varying from 8.6 to 19.8 days. Lethal Concentrations (LC50 of 3.93 × 10(6 conidia mL-1 and 7.44 × 10(8 conidia mL-1 were determined for B. bassiana and M. anisopliae, respectively. B. bassiana isolate 645 was the most efficient, being an alternative for use in biological control programs against the cassava green mite.

  3. Isolation and efficacy of entomopathogenic fungus (Metarhizium anisopliae) for the control of Aedes albopictus Skuse larvae:suspected dengue vector in Pakistan

    Institute of Scientific and Technical Information of China (English)

    Hazrat Bilal; Soaib Ali Hassan; Imtinan Akram Khan

    2012-01-01

    Objective: To isolate the entomopathogenic fungus Metarhizium anisopliae (M. anisopliae) in the local environment, and evaluate its efficacy against the suspected dengue vector Aedes albopictus in Pakistan. Methods: According to the standard procedure, M. anisopliae was isolated from the dead mosquitoes which were collected from the field or dead after the collection. Bioassay was performed to determine its efficacy. Results: The results indicated that M. anisopliae had larvicidal effect with LC50 value 1.09×105 and LC90 value 1.90×1013 while it took 45.41 h to kill 50%of tested population. Conclusions: Taking long time to kill 50% population when compare with the synthetic insecticides, is the only drawback for the use of entomopathogenic fungus but these bio-pesticides are safe for the use.

  4. Monitoring persistence of the entomopathogenic fungus Metarhizium anisopliae under simulated field conditions with the aim of controlling adult Aedes aegypti (Diptera: Culicidae).

    Science.gov (United States)

    Carolino, Aline T; Paula, Adriano R; Silva, Carlos P; Butt, Tariq M; Samuels, Richard I

    2014-04-25

    Entomopathogenic fungi are potential candidates for use in integrated vector management, with recent emphasis aimed at developing adult mosquito control methods. Here we investigated the persistence of the fungus Metarhizium anisopliae when tested against female A. aegypti under field conditions. Black cotton cloths impregnated with M. anisopliae conidia, formulated in vegetable oil + isoparaffin, were maintained on a covered veranda for up to 30 days. At specific times, pieces of the cloths were removed, placed in Tween 80 and the resuspended conidia were sprayed directly onto mosquitoes. The persistence of conidia impregnated on black cloths using three different carriers was evaluated in test rooms. Fifty mosquitoes were released into each room and after a 5 day period, the surviving insects were captured. Another 50 insects were then released into each room. The capacity of the fungus at reducing mosquito survival was evaluated over a total of 35 days. Conidia extracted from cloths maintained on the veranda for 2 to 18 days remained virulent, with 28 to 60% mosquito survival observed. Mosquito survival following exposure to fungus impregnated cloths showed that fungus + Tween caused similar reductions to that of fungus + vegetable oil. Mosquitoes exposed to the formulation fungus + vegetable oil had survival rates of 36% over the first 5 days of the experiment. Following the release of the second cohort of mosquitoes (6-11days), survival increased to 50%. The survival of the 12-17 day cohort (78%) was statistically equal to that of the controls (84%). Formulation of the fungus in vegetable oil + isoparaffin increased the persistence of the fungus, with the 18-23 day cohort (64% survival) still showing statistical differences to that of the controls (87% survival). The potential of entomopathogenic fungi for the control of adult A. aegypti was confirmed under field conditions. Vegetable oil + isoparaffin formulations of M. anisopliae significantly increased the

  5. Effectiveness of Metarhizium anisopliae against immature stages of Anastrepha fraterculus fruitfly (Diptera : Tephritidae Eficácia de Metarhizium anisopliae a estágios imaturos de Anastrepha fraterculus a mosca das frutas (Diptera : Tephritidae

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    Ricardo Henry Rodrigues Destéfano

    2005-03-01

    Full Text Available The study evaluated the effectiveness of Metarhizium anisopliae var. anisopliae (Hyphomycetes : Moniliales strain E9, isolated from the pasture spittlebug Deois flavopicta (Hemiptera : Cercopidae, against larvae, prepupae and pupae stage and emergent adults of Anastrepha fraterculus, the South American fruitfly. The bioassay was carried out simulating field conditions, on autoclaved (AS and non-autoclaved (NAS soil from typical citrus orchards in the State of São Paulo, Southeastern region of Brazil. Various concentrations of conidia were incorporated into the soil the mortality, calculated based on the percentage of adult emergence, was 86% for the highest conidia concentrations: 2.52 x 10(10 for AS and 2.52 x 10(10 for NAS. The lethal concentration (LC50, expressed as conidia concentration, was 8.44 x 10(9 conidia/g of soil (S for AS and 12.23 x 10(9 conidia/g of soil for NAS.O estudo avaliou a eficácia de Metarhizium anisopliae var. anisopliae, (Hyphomycetes : Moniliales linhagem E9, isolada da cigarrinha das pastagens Deois flavopicta (Hemiptera : Cercopidae, contra larvas, prepupas, pupas e adultos emergentes de Anastreha fraterculus, a mosca Sul Americana das frutas. Os bioensaios foram conduzidos simulando condições de campo em solo autoclavado (AS e não autoclavado (NAS de pomares típicos de citros no Estado de São Paulo, sudeste do Brasil. Várias concentrações de conídios foram incorporadas no solo. A mortalidade calculada sobre a porcentagem de adultos emergentes, foi de 86% para as concentrações mais altas de conídios: 2,52 x 10(10 para AS e 2,52 x 10(10 para NAS. A concentração letal (LC50 expressa pela concentração de conídios, foi 8,44 x 10(9 conídios/g de solo para AS e 12,2 x 10(9 conídios/g/S para NAS.

  6. Identification of a hybrid PKS-NRPS required for the biosynthesis of NG-391 in Metarhizium anisopliae var. anisopliae

    Science.gov (United States)

    A 19,818 kb genomic region harboring six predicted ORFs was identified in M. anisopliae ARSEF 2575. ORF4, putatively encoding a hybrid polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS) was targeted using Agrobacterium-mediated gene knockout. Homologous recombinants failed to produce det...

  7. Production of destruxins from metarhizium spp. fungi in artificial medium and in endophytically colonized cowpea plants

    Science.gov (United States)

    Destruxins (DTXs) are cyclic depsipeptides produced by many Metarhizium isolates that have long been assumed to contribute to virulence of these entomopathogenic fungi. We evaluated the virulence of 20 Metarhizium isolates against insect larvae and measured the concentration of DTXs A, B, and E prod...

  8. Clarification of generic and species boundaries for Metarhizium and related fungi through multigene phylogenetics

    Science.gov (United States)

    The genus Metarhizium traditionally refers to green-spored asexual insect pathogenic fungi. Through culturing and molecular methods, Metarhizium has been linked to Metacordyceps sexual states. Historically, fungal nomenclature has allowed separate names for the different life-stages of pleomorphic...

  9. New biotypes of Metarhizium anisopliae var. anisopliae E9 strain with altered conidial germination, obtained by exposition to gamma radiation; Novos biotipos de Metarhizium anisopliae var. anisopliae (Metsch.) Sorokin com germinacao alterada de conidios, obtidos pela exposicao a radiacao gama

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, M.G.; Oliveira, N.T.; Luna Alves Lima, E.A. [Pernambuco Univ., Recife, PE (Brazil). Centro de Ciencias Biologicas. Dept. de Micologia

    1997-12-31

    Conidia produced by a wild strain (E9) of the entomopathogenic fungus M. anisopliae var anisopliae were exposed to gamma radiation in order to obtain new biotypes. At the 390 Gy dose there were obtained 48 colonies (MaE). On complete medium, 5 colonies (MaE 01, MaE 10, MaE 15, MaE 40) presented morphological changes in color while the colony MaE 24 lost its esporulation capacity. Twenty six colonies presented mycelial growth significantly different from the wild strain, after 12 days of incubation. Twelve colonies showed average of conidial germination different from the wild strain, after 12 days of incubation on liquid minimum medium at 25 deg C. The colony MaE started germination precociously after 5 hours of incubation. (author) 31 refs., 1 fig., 4 tabs.

  10. Selection of Metarhizium anisopliae (Metsch. and Beauveria bassiana (Bals. isolates to control Alabama argillacea (Huebner caterpillars Seleção de isolados de Metarhizium anisopliae (Metsch. e Beauveria bassiana (Bals. para o controle de lagartas de Alabama argillacea (Huebner

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    Everardo César Filho

    2002-09-01

    Full Text Available The cotton leafworm, Alabama argillacea (Huebner, 1818 (Lepidoptera: Noctuidae, is considered to be one of the key pests in herbaceous cotton (Gossypium hirsutum L. r. latifolium Hutch cropping, with constant occurrence in all cotton-growing states of Brazil. In this study Metarhizium anisopliae and Beauveria bassiana isolates were screened and evaluated for pathogenicity against Alabama argillaceae. Initially, a screening of ten isolates of each fungus in a concentracion of 10(8 conidia mL-1, was carried out on 3rd instar larvae of A. argillacea. Further studies were conducted to determine the pathogenicity and virulence of six and seven isolates of M. anisopliae and B. bassiana, respectively, against 3rd instar larvae of A. argillacea and using the concentrations of 10(6, 10(7, 10(8, and 10(9 conidia mL-1. The experiments were carried out in Recife, PE, Brazil, at 27 ± 2ºC, RH 70 ± 5% and a photophase of 12 hours. Mortalities caused by M. anisopliae isolate at the different concentrations ranged from 4.5 to 91.2%, the highest mortality percentage being found for the isolate 1189 at 10(9 conidia mL-1. The isolate 645 of B. bassiana caused the highest mortality at the highest concentration, followed by isolates 634, 604, and IPA 198. The lowest lethal time for B. bassiana and M. anisopliae, was achieved by the isolates 483 (4.1 days and 1189 (2.0 days, respectively. The isolates 1189, 1022 e 866 of M. anisopliae and 483, IPA198 and 604 of B. bassiana, at 10(8 e 10(9 conidia mL-1 are promissing for use the integrated control of A. argillacea larvae, but M. anisopliae seems more effective.O curuquerê-do-algodoeiro (Alabama argillacea é considerado uma das principais pragas do algodoeiro herbáceio no Brasil, com ocorrência comum em vários estágios de crescimento da cultura. Este trabalho avaliou a patogenicidade e selecionou isolados dos fungos entomopatogênicos Metarhizium anisopliae e Beauveria bassiana, para lagartas do curuquer

  11. Integrated control of an acaricide-resistant strain of the cattle tick Rhipicephalus microplus by applying Metarhizium anisopliae associated with cypermethrin and chlorpyriphos under field conditions.

    Science.gov (United States)

    Webster, Anelise; Reck, José; Santi, Lucélia; Souza, Ugo A; Dall'Agnol, Bruno; Klafke, Guilherme M; Beys-da-Silva, Walter O; Martins, João Ricardo; Schrank, Augusto

    2015-01-30

    The efficacy of the fungus Metarhizium anisopliae to control ticks has been shown in several in vitro experiments. However, few studies have been undertaken in field conditions in order to demonstrate the applicability of its use as a biological control of ticks and its combination with chemical acaricides. The aim of the present study was to evaluate the efficacy of M. anisopliae to control an acaricide-resistant strain of Rhipicephalus microplus under laboratory and field conditions. First, the compatibility of M. anisopliae strain (TIS-BR03) with commercial acaricides and its potential to control the cattle tick were evaluated in vitro. In general, acaricide treatments had mild effects on fungus viability. In the field experiment, the median of treatment efficacy with acaricide only, M. anisopliae only and combination of M. anisopliae with acaricide were 71.1%, 56.3% and 97.9%, respectively. There is no statistical difference between groups treated with M. anisopliae and acaricide alone. Thus, in this work we have demonstrated the applicability of M. anisopliae use associated or not with chemical acaricides on field conditions in order to control an acaricide-resistant strain of the cattle tick R. microplus.

  12. Efeito de Beauveria bassiana (Bals. Vuill. e Metarhizium anisopliae (Metsch. Sorok. sobre características biológicas de Diatraea saccharalis F. (Lepidoptera: Crambidae = Effect of Beauveria bassiana (Bals. Vuill. and Metarhizium anisopliae (Metsch. Sorok. on Biological Characteristics of Diatraea saccharalis F. (Lepidoptera: Crambidae

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    Marco Aurélio Paes de Oliveira

    2008-04-01

    Full Text Available A broca da cana-de-açúcar Diatraea saccharalis F. é considerada uma das principais pragas nas Américas. Entre os métodos de controle, o uso de fungos entomopatogênicos tem sido amplamente recomendado no manejo das pragas da cana-de-açúcar, incluindo outras lepidobrocas. Assim sendo, este estudo investigou os efeitos de diferentes concentrações de Beauveria bassiana (Bals. Vuill. e Metarhizium anisopliae (Metsch. Sorok. sobre parâmetros biológicos da broca da cana-de-açúcar. Larvas de terceiro instar de D. saccharalis foram tratadas com os fungos usando as concentrações de 103, 104 e 105 conídios mL-1. Larvas tratadas com 105 conídios mL-1 de B. bassiana tiveram menor sobrevivência (56,6%, comparadas com lagartas não-tratadas (90%. Adultos originados de larvas tratadas colocaram menor número de ovos, com menor viabilidade, e viveram menos, comparados com adultos originados de larvas não-tratadas. Larvas tratadas com M. anisopliae na concentração de 105 conídios mL-1 e adultos originados destas larvas também exibiram redução no desempenho, comparados aos insetos não-tratados. Os resultados indicam que B. bassiana e M. anisopliae, além de patogênicos àslarvas de D. saccharalis, também interferem negativamente na sua biologia, mostrando potencial de uso contra esta praga.The sugarcane borer Diatraea saccharalis F. is considered oneof the major sugarcane pests in the American continent. Among control methods, the use of entomopathogenic fungi has been broadly recommended to manage sugarcane pests, including other sugarcane borers. Therefore, this study investigated the effects of differentconcentrations of Beauveria bassiana (Bals. Vuill and Metarhizium anisopliae (Metch. Sorok on biological characteristics of the sugarcane borer. Third-instar larvae of D. saccharalis werefungi-treated using the concentrations of 103, 104 and 105 conidia mL-1. Larvae treated with 105 conidia mL-1 of B. bassiana showed lower survival

  13. Screening of Metarhizium anisopliae (Metsch. Sorok. strains against Mahanarva fimbriolata (Stal, 1854 (Hemiptera: Cercopidae/ Seleção de isolados de Metarhizium anisopliae (Metsch. Sorok. a Mahanarva fimbriolata (Stal, 1854 (Hemiptera: Cercopidae

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    Solange Aparecida Vieira

    2006-06-01

    Full Text Available The mechanized harvest of sugarcane results in accumulation of mulch on the soil and promotes favorable temperature and moist to development of sugarcane root spittlebug, Mahanarva fimbriolata (Stal, 1854 (Hemiptera: Cercopidae. High populations of this pest causes serious damages to plants and high costs with insecticides. This research work tested strains of Metarhizium anisopliae (Metsch. Sorok. pathogenic to sugarcane root spittlebug using a new methodology. The selection was carried out in laboratory conditions. Nymphs were sprayed with 2 mL of a suspension containing 5x107 conidia/mL and maintained in roots of sugarcane. The corrected mortality was evaluated five days after pulverization and ranged from 10.5 to 60% and the more pathogenic strains caused mortality from 46.5 to 59.7%.A colheita mecanizada de cana-de-açúcar, sem a queima da palhada, proporciona acúmulo de matériaorgânica no solo tornando o ambiente favorável ao desenvolvimento da cigarrinha-da-raiz, Mahanarva fimbriolata (Stal, 1854 (Hemiptera: Cercopidae. Em elevadas infestações essa praga tem causado prejuízos devido aos grandes danos nas plantas e aos gastos com agrotóxicos. O objetivo da pesquisa foi selecionar isolados de Metarhizium anisopliae (Metsch. Sorok. patogênicos para cigarrinha-da-raiz, usando uma nova metodologia. A seleção foi feita em condições de laboratório, utilizando-se ninfas coletadas a campo que foram pulverizadas com o fungo e mantidas em raiz de mudas de cana-de-açúcar. A mortalidade corrigida ao quinto dia após a inoculação variou de 10,5 a 60%, sendo que os isolados mais patogênicos causaram mortalidades de 46,5 a 59,7%.

  14. Purification and characterisation of proteins secreted by the entomopathogenic fungus Metarhizium anisopliae with insecticidal activity against adults of the Mediterranean fruit fly, Ceratitis capitata (Diptera: Tephritidae).

    Science.gov (United States)

    Ortiz-Urquiza, Almudena; Garrido-Jurado, Inmaculada; Santiago-Alvarez, Cándido; Quesada-Moraga, Enrique

    2009-10-01

    The control of the Mediterranean fruit fly (medfly) Ceratitis capitata (Wied) is usually performed with protein bait sprays incorporating chemical insecticides that may have adverse effects on humans, non-target organisms and the environment. In recent years, scientists have sought more environmentally friendly insecticides for medfly control, such as plant- and microorganism-derived compounds. Among these compounds, entomopathogenic fungi are an unexplored source of natural insecticides. The crude soluble protein extract (CSPE) of the entomopathogenic fungus Metarhizium anisopliae (Mestch.) (strain EAMa 01/58-Su) shows chronic insecticidal activity when administered per os. Mortality in flies exhibits a dose response. The CSPE produces an antifeedant effect in adult flies, a result probably due to a progressive deterioration of the fly midgut after ingestion of the extract. Protease and temperature treatments show that insecticidal activity against C. capitata is due to proteinaceous compounds that are highly thermostable. Four monomeric proteins from this crude extract have been purified by liquid chromatography and gel electroelution. Although all four monomers seem to be involved in the insecticidal activity of the CSPE, the 15 kDa and the 11 kDa proteins appear to be mainly responsible for the observed insecticidal effect. Four new fungal proteins with insecticidal activity have been purified and identified. These proteins might be combined with insect baits for C. capitata biocontrol. Copyright 2009 Society of Chemical Industry.

  15. Mecanismo de infecção do fungo Metarhizium anisopliae no carrapato Boophilus microplus em condições experimentais The penetration of the fungus Metarhizium anisopliae on Boophilus microplus in experimental conditions

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    Vânia Rita Elias Pinheiro Bittencourt

    1999-06-01

    Full Text Available O objetivo do presente trabalho foi avaliar o mecanismo de penetração do fungo Metarhizium anisopliae em Boophilus microplus em condições experimentais. As infecções foram realizadas utilizando amostra de M. anisopliae isolada de carrapatos. A suspensão de conídios do fungo foi preparada utilizando água destilada e espalhante adesivo Tween 80 e foi quantificada na concentração de 10(8 conídios/ml. As fêmeas ingurgitadas foram banhadas nesta suspensão durante cinco minutos e, após este período, foram levadas à câmara climatizada. Três dias após a infecção, as fêmeas foram fixadas utilizando tetróxido de ósmio e glutaraldeído, posteriormente desidratadas em bateria de álcool etílico com acetona. Após a desidratação, esse material foi levado à metalização e ao microscópio eletrônico de varredura, onde se observou a fixação dos conídios na cutícula das fêmeas, havendo germinação e a dilatação da extremidade do tubo germinativo em todas as amostras. Esta dilatação observada foi devido à formação do apressório, apesar de não ser possível observar esta estrutura sobre a cutícula das fêmeas ingurgitadas. O tetróxido de ósmio mostrou ser um excelente fixador para fungos, enquanto o glutaraldeído mostrou ser eficiente na fixação do artrópode. Conclui-se que a forma principal de penetração deste entomopatógeno em B. microplus é através da cutícula. Cabe aqui frisar que esta é a primeira vez que é descrito o mecanismo de penetração de M. anisopliae em carrapatos.The objective of the present communication has been to draw attention to the mode of penetration of the fungus Metarhizium anisopliae through the cuticle of Boophilus microplus under experimental conditions. Samples of M. anisopliae isolated from naturally infected ticks were used to prepare a suspension of 108 conidia/ml in a destilled water with 2% of Tween 80. The engorged females were submergged in this suspension for five

  16. Real encoded genetic algorithm and response surface methodology to optimize production of an indolizidine alkaloid, swainsonine, from Metarhizium anisopliae.

    Science.gov (United States)

    Singh, Digar; Kaur, Gurvinder

    2013-09-01

    Response surface methodology (RSM) and artificial neural network-real encoded genetic algorithm (ANN-REGA) were employed to develop a process for fermentative swainsonine production from Metarhizium anisopliae (ARSEF 1724). The effect of finally screened process variables viz. inoculum size, oatmeal extract, glucose, and CaCl2 were investigated through central composite design and were further utilized for training sets in ANN with training and test R values of 0.99 and 0.94, respectively. ANN-REGA was finally employed to simulate the predictive swainsonine production with best evolved media composition. ANN-REGA predicted a more precise fermentation model with 103 % (shake flask) increase in alkaloid production compared to 75.62 % (shake flask) obtained with RSM model upon validation.

  17. Evaluating physical and nutritional stress during mycelial growth as inducers of tolerance to heat and UV-B radiation in Metarhizium anisopliae conidia.

    Science.gov (United States)

    Rangel, Drauzio E N; Anderson, Anne J; Roberts, Donald W

    2008-11-01

    Elevated tolerance to UV-B radiation and heat may be induced in conidia produced on fungi exposed during mycelial growth to sublethal stresses other than heat or UV-B. This is due to a phenomenon referred to as 'cross-protection'. Several mechanisms are associated with this increased conidial tolerance, one of which is the accumulation of trehalose and mannitol within conidia. In the present study, conidia of the insect-pathogenic fungus Metarhizium anisopliae var. anisopliae were produced on mycelium subjected to nutritive, heat-shock, osmotic, or oxidative stress. The tolerance levels to UV-B radiation and heat of the conidia from stressed mycelium were evaluated, and the amounts of trehalose and mannitol accumulated in conidia were quantified. Conidia produced under nutritive stress (carbon and nitrogen starvation) were two-times more heat and UV-B tolerant than conidia produced under rich (non-stress) nutrient conditions [potato-dextrose agar with yeast extract (PDAY)], and they also accumulated the highest concentrations of trehalose and mannitol. Conidia produced on heat-shock stressed PDAY cultures had higher tolerance to UV-B radiation and heat than conidia produced without heat shock; however, both the UV-B tolerance and trehalose/mannitol concentrations in conidia produced on heat-shocked mycelium were less than those of conidia produced under nutritive stress. Conidia produced under osmotic stress (sodium or potassium chloride added to PDAY) had elevated heat and UV-B tolerances similar to those of conidia produced under nutritive stress; however, they had the lowest levels of mannitol and trehalose, which indicates that accumulation of these compounds is not the only mechanism used by M. anisopliae for protection from heat and UV-B radiation. Oxidative stress from UV-A irradiation or hydrogen peroxide did not produce conidia with elevated UV-B or heat tolerances. Conidia produced under oxidative stress generated by menadione had increased or unchanged

  18. Density, Viability Conidia And Symptoms of Metarhizium anisopliae infection on Oryctes rhinoceros larvae

    Science.gov (United States)

    Indriyanti, D. R.; Putri, R. I. P.; Widiyaningrum, P.; Herlina, L.

    2017-04-01

    M. anisopliae is parasitic fungus on insect pests; it is used as a biocontrol agent. M. anisopliae can be propagated on maize or rice substrate. M. anisopliae is currently sold in the form of kaolin powder formulations. Before it is used to check the density, viability and pathogenicity of M. anisopliae. However the problem is the kaolin powder very soft, so it difficult to distinguish between kaolin and conidia. This article gives information on how to calculate conidia density, viability and symptoms of M. anisopliae infection on Oryctes rhinoceros larvae. The study was conducted in the laboratory to determine the density and viability. The pathogenicity testing was done using pots. The Pot is containing soil substrate mixed with M. Anispoliae and ten tails O. Rhinoceros larvae per pot. The results showed that the density of M. anisopliae conidia was 1.81 x 108 conidia mL-1 and the viability was 94% within 24 hours. The larval mortality began to emerge in the 1st week, and all larvae died at the sixth week. The symptom of M. anisopliae infection on Oryctes rhinoceros larvae, there was a black spot on the larval integument. The larvae movements become slow and poor appetite; it will die within 3-7 days. The larvae die hard, and the white hyphae grow on the body surface that turns green.

  19. Enhanced ovicidal activity of an oil formulation of the fungus Metarhizium anisopliae on the mosquito Aedes aegypti.

    Science.gov (United States)

    Albernaz, D A S; Tai, M H H; Luz, C

    2009-06-01

    The effect of humidity on the activity of Metarhizium anisopliae IP 46 (Metsch.) Sorokin (Hypocreales: Clavicipitaceae) formulated in sunflower oil against Aedes aegypti (L.) (Diptera: Culicidae) eggs was examined. After exposure of eggs at 75% relative humidity (RH) for ovicidal activity was not increased by oil-in-water formulated conidia, hyphal bodies or pure-oil formulated conidia, compared with conidia or hyphal bodies prepared in water only. At optimal > 98% RH, eclosion was Ovicidal activity was still detected at 93% RH and was augmented with increasing humidity and time of exposure of eggs. Eclosion of larvae was distinctly reduced by IP 46 pure-oil formulated conidia after a minimal initial exposure of 3 days at > 98% RH, followed by: (a) a 12-day exposure at 75% RH before submersion in water; (b) a minimal 5-day exposure at > 98% RH and direct subsequent transfer of treated eggs to water, or (c) a minimal daily 20-h exposure at > 98% RH alternating with 4 h at 75% RH for 10 days. We demonstrate that oil-based formulations of conidia of M. anisopliae enhance ovicidal activity at high humidities and conclude that these formulations have potential in the integrated control of Ae. aegypti.

  20. Horizontal Transmission of the Entomopathogen Fungus Metarhizium anisopliae in Microcerotermes diversus Groups

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    Mohammad Saied Mossadegh

    2012-08-01

    Full Text Available An experiment was carried out in order to investigate fungal conidia transmission of Metarhizium anisopliae (Metschnikoff Sorokin from vector (donor to healthy Microcerotermes diversus Silvestri (Iso.: Termitidae and determine the best donor/concentration ratio for transmission. After preliminary trials, concentrations of 3.1 × 104, 3.9 × 105, 3.2 × 106 and 3.5 × 108 conidia mL−1 were selected for testing. The experiment was performed at three donor : Recipient ratios of 10, 30 and 50%. The highest mortality of recipient workers was observed after 14 days at the concentration of 3.5 × 108 conidia mL−1 and donor ratio of 50%. The mortality of recipient workers was less than 20% at all concentrations at a donor ratio of 10%. Our observations indicate social behavior of M. diversus, such as grooming, can be effective in promoting epizootic outbreaks in a colony. While the current results suggest good potential for efficacy, the use of M. anisopliae as a component of integrated pest management of M. diversus still needs to be proven under field conditions.

  1. Allergic Responses Induced by a Fungal Biopesticide Metarhizium anisopliae and House Dust Mite Are Compared in a Mouse Model

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    Marsha D. W. Ward

    2011-01-01

    Full Text Available Biopesticides can be effective in controlling their target pest. However, research regarding allergenicity and asthma development is limited. We compared the ability of fungal biopesticide Metarhizium anisopliae (MACA and house dust mite (HDM extracts to induce allergic responses in BALB/c mice. The extracts were administered by intratracheal aspiration at doubling doses (2.5–80 g protein 4X over a four-week period. Three days after the last exposure, serum and bronchoalveolar lavage fluid (BALF were collected. The extracts' relative allergenicity was evaluated based on response robustness (lowest significant dose response compared to control (0 g. MACA induced a more robust serum total IgE response than HDM. However, in the antigen-specific IgE assay, a similar dose of both MACA and HDM was required to achieve the same response level. Our data suggest a threshold dose of MACA for allergy induction and that M. anisopliae may be similar to HDM in allergy induction potential.

  2. 绿僵菌防治蛴螬的应用研究进展%Research progress in application of Metarhizium anisopliae against white grubs

    Institute of Scientific and Technical Information of China (English)

    于永浩; 龙秀珍; 曾宪儒; 韦德卫; 曾涛

    2013-01-01

    White grub is an important global underground pest, which could be effectively controlled by Metarhizium anisopliae, a crucial entomopathogenic fungus for effective insect pest control. This paper reviewed the domestic and international researches on Metarhizium anisopliae control against white grubs and the development and application status of Metarhizium anisopliae commercial formulation; at the same time, this paper also discussed the influences of temperature, humidity, ultraviolet radiation, and other environmental factors on the effects of Metarhiziutn anisopliae on white grubs control. In conclusion, this research report pointed out studies on AIDS screening, R&D mixture preparation, strain degradation mechanism, and soil antibacterial action in order to provide research focuses and directions for Metarhizium anisoplian control against white grubs.%蛴螬是重要的世界性地下害虫,绿僵菌是防治害虫的重要昆虫病原真菌,对蛴螬有良好的防治效果.文章综述了国内外对绿僵菌防治蛴螬的研究报道及商品制剂的研制和应用现状,讨论了温湿度和紫外线等环境因素对绿僵菌防治蛴螬效果的影响;指出助剂的筛选、混合制剂的研发、菌株退化机理和土壤抑菌作用的研究等是绿僵菌防治蛴螬的研究热点和方向.

  3. Utilization of metarhizium anisopliae and vegetable extract for control of the spodoptera frugiperda and helicoverpa zea (lepidoptera: noctuidae in the corn

    Directory of Open Access Journals (Sweden)

    Eduardo Barbosa Beserra

    2008-05-01

    Full Text Available The potential use of the vegetable extracts and Metarhizium anisopliae about the population and damage of the Spodoptera frugiperda and Helicoverpa zea in corn, it was evaluated when planted in two spacings. The experiment was installed in the culture of corn, in the city of Lagoa Seca/PB, in two areas of 500 m2, in the spacings 0,8 x 0,4 and 0,4 x 0,4 m, with two and one plant for hole. The basis was constituited of: 1 Water as witness; 2 Metarhizium anisopliae; 3 Extracts of Mormodica charantia; 4 Metarhizium anisopliae + extracts of M. charantia; 5 M. anisopliae + manipueira. The results showed some statistical difference between the two spacings, with a larger number of collected caterpillars, 5,84, and a larger damage to the plant, 3,24, in the spacing 0,4 x 0,4 m, in relation to the spacing 0,8 x 0,4 m, within as average of 4,13 caterpillars and danificated of 2,71. There was not any significant interaction between the spacings and the products used for the control of S. frugiperda. However, there was a significant difference among the treatments, with a smaller number of caterpillars, 4,17, and a smaller medium damage to the plant, 2,81, when was used, M. anisopliae + extract of M. charantia. Considering to the control of the plaque the percentage of damage ears of corn by H. zea, weight and length of the ear of corn in corn, there wasn’t neither significant difference with regard to the spacing, neither with regard to the used treatments.

  4. Human sera IgE reacts with a Metarhizium anisopliae fungal catalase

    Science.gov (United States)

    Background: Previous studies have demonstrated that Metarhzium anisopliae extract can induce immune responses in a mouse model that are characteristic of human allergic asthma. Objectives: The objective of this study was to identify and characterize the extract proteins t...

  5. Evaluation of Metarhizium anisopliae for the control of Culicoides brevitarsis Kieffer (Diptera: Ceratopogonidae), the principal vector of bluetongue virus in Australia.

    Science.gov (United States)

    Nicholas, A H; McCorkell, B

    2014-06-01

    Four isolates of the entomopathogenic fungus Metarhizium anisopliae were tested for their potential to control the biting midge Culicoides brevitarsis, the principal vector of bluetongue virus in Australia. Adult C. brevitarsis died three to eight days after walking on paper substrate treated with 0.7 g/m(2) conidia of any of the isolates, indicating that M. anisopliae has potential as a surface treatment or topical application control strategy. Incorporation of the fungus into freshly excreted cattle dung at rates of between 0.25 and 1 g conidia/kg reduced the emergence of adult midges by up to 98.5% compared to untreated dung indicating that M. anisopliae has the potential to control C. brevitarsis larvae in cattle dung. Three of the isolates produced similar mortality rates on adult and immature C. brevitarsis while the fourth isolate produced lower, but still significant, mortality rates on adult and immature stages.

  6. Protease production during growth and autolysis of submerged Metarhizium anisopliae cultures Produção de protease durante o crescimento e análise de culturas submersas de Metarhizium anisopliae

    Directory of Open Access Journals (Sweden)

    Gilberto U.L. Braga

    1999-04-01

    Full Text Available The growth and autolysis of two strains of the entomopathogenic deuteromycete fungus Metarhizium anisopliae var. anisopliae were evaluated in medium containing casein or glucose as carbon source. Parameters such as economic coefficient and degree of autolysis were determined for each strain. Protease production was determined throughout the growth and autolysis phases of the cultures on medium under conditions of protease induction (in the presence of casein as sole source of carbon and nitrogen. The fungus was shown to utilize casein as a carbon/energy source in a more efficient manner than glucose. The autolysis shown by the strains was intense under both types of growth conditions, reaching up to 62.7% of the dry mass produced and started soon after the depletion of the exogenous carbon source. The relationship between the proteolytic activities of the two strains evaluated varied significantly (a maximum of 19.78 on the 5th day and a minimum of 2.03 on the 16th day of growth during the various growth and autolysis phases, clearly showing that the difference between the growth curves and the difference in the kinetics of enzyme production may decisively affect the process of strain selection for protease production.O crescimento e a autólise de duas linhagens do deuteromiceto entomopatogênico Metarhizium anisopliae var. anisopliae foram avaliados em meio contendo caseína ou glicose como fonte de carbono. Foram determinados parâmetros como o coeficiente econômico e o grau de autólise apresentado pelas linhagens. A produção de protease foi determinada durante todas as fases do crescimento e da autólise das culturas, em meio indutor da produção de proteases (meio contendo caseína como única fonte de carbono e de nitrogênio. Pôde-se verificar que o fungo foi capaz de utilizar a caseína como fonte de carbono/energia de maneira mais eficiente do que a glicose. A autólise apresentada pelas linhagens foi intensa em ambas as condi

  7. Efeito de beauveria bassiana (bals. Vuillemin e Metarhizium anisopliae (metsch. sorokin nos parâmetros biológicos de trichogramma atopovirilia oatman & platner, 1983 (hymenoptera: trichogrammatidae Effect of Beauveria bassiana (Bals. Vuillemin and Metarhizium anisopliae (Metsch. Sorokin on the biological parameters of Trichogramma atopovirilia Oatman & Platner (Hymenoptera: Trichogrammatidae

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    Ricardo Antonio Polanczyk

    2010-12-01

    Full Text Available Dois experimentos foram realizados para avaliar o efeito de duas formulações comerciais à base de Beauveria bassiana (Bals. Vuillemin e Metarhizium anisopliae (Metsch., Sorokin sobre os parâmetros biológicos de Trichogramma atopovirilia Oatman & Platner, 1983. No primeiro experimento, cartelas com ovos de Spodoptera frugiperda, 1797 foram mergulhadas em suspensões preparadas com os produtos e, em seguida, colocadas à disposição dos parasitóides para oviposição, durante um período de 24 horas. No segundo, fêmeas do parasitóide foram alimentadas com uma solução mel e suspensão de conídios. Em ambos os experimentos as fêmeas mortas foram colocadas em uma câmara úmida para observar a esporulação dos fungos. Os tratamentos foram mantidos em câmara climatizada com temperatura de 25±1º C, umidade relativa de 70±10% e fotofase de 14 horas. Avaliou-se a longevidade e mortalidade dos adultos, índice de parasitismo, emergência do parasitóide, número de indivíduos por ovo e razão sexual dos descendentes. As formulações não interferiram nos parâmetros avaliados e não foi observada a esporulação do fungo no cadáver do parasitóide adulto. É possível inferir que T. atopovirilia e os fungos entomopatogênicos B. bassiana e M. anisopliae são compatíveis e podem ser empregados simultaneamente em programas de manejo integrado de S. frugiperda.Two bioassays were performed to evaluate the effect of two biopesticides based on Beauveria bassiana (Bals. Vuillemin and Metarhizium anisopliae (Metsch. Sorokin on the biological parameters of Trichogramma atopovirilia Oatman & Platner, 1983. In the first one, displays with S. frugiperda, 1797 eggs were dropped into the biopesticide suspension and offered to the parasitoid females for 24 hours. In the second one, parasitoid females were fed with a suspension containing honey and biopesticide suspension. In both cases, after the parasitoid death they were mantained into a humid

  8. Vectorial capacity of Aedes aegypti for dengue virus type 2 is reduced with co-infection of Metarhizium anisopliae.

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    Javier A Garza-Hernández

    Full Text Available BACKGROUND: Aedes aegypti, is the major dengue vector and a worldwide public health threat combated basically by chemical insecticides. In this study, the vectorial competence of Ae. aegypti co-infected with a mildly virulent Metarhizium anisopliae and fed with blood infected with the DENV-2 virus, was examined. METHODOLOGY/PRINCIPAL FINDINGS: The study encompassed three bioassays (B. In B1 the median lethal time (LT50 of Ae. aegypti exposed to M. anisopliae was determined in four treatments: co-infected (CI, single-fungus infection (SF, single-virus infection (SV and control (C. In B2, the mortality and viral infection rate in midgut and in head were registered in fifty females of CI and in SV. In B3, the same treatments as in B1 but with females separated individually were tested to evaluate the effect on fecundity and gonotrophic cycle length. Survival in CI and SF females was 70% shorter than the one of those in SV and control. Overall viral infection rate in CI and SV were 76 and 84% but the mortality at day six post-infection was 78% (54% infected and 6% respectively. Survivors with virus in head at day seven post-infection were 12 and 64% in both CI and SV mosquitoes. Fecundity and gonotrophic cycle length were reduced in 52 and 40% in CI compared to the ones in control. CONCLUSION/SIGNIFICANCE: Fungus-induced mortality for the CI group was 78%. Of the survivors, 12% (6/50 could potentially transmit DENV-2, as opposed to 64% (32/50 of the SV group, meaning a 5-fold reduction in the number of infective mosquitoes. This is the first report on a fungus that reduces the vectorial capacity of Ae. aegypti infected with the DENV-2 virus.

  9. A protocol for determination of conidial viability of the fungal entomopathogens Beauveria bassiana and Metarhizium anisopliae from commercial products.

    Science.gov (United States)

    Oliveira, Daian Guilherme Pinto; Pauli, Giuliano; Mascarin, Gabriel Moura; Delalibera, Italo

    2015-12-01

    Techniques for directly determining conidial viability have widespread use but also have limitations for quality control assessments of formulated mycoinsecticides, especially in emulsifiable oil. This study proposes a new method based on adaptations of already established protocols that use the direct viability method to make it more economical and accurate, thus enabling its use in the evaluation of formulated products. Appropriate parameters and conditions were defined using products based on Beauveria bassiana and Metarhizium anisopliae in the forms of pure conidia, fungus-colonized rice, ground fungus-colonized rice and oil dispersion. The established protocol, named ESALQ, consists of the transfer of 150 μL of a suspension containing about 0.7 and 1 × 10(6) conidia/mL onto Rodac® plates with 5 mL of potato dextrose agar culture medium + 5 mg/L of Pentabiotic® and 10 μL/L of Derosal® (Carbendazim) and subsequent counting of germinated and non-germinated conidia. For the ground fungus-colonized rice and oil dispersion formulations, prior to transferring the fungal suspension to the medium, rice should be decanted and the oil removed, respectively. This method was compared with another direct viability method and with the Colony-forming unit (CFU) and Fluorescence viability methods, comparing the accuracy obtained using the coefficient of variation (CV) of the analysis of each method. The results showed that in addition to the ease of application, the developed method has higher accuracy than the other methods (with a CV up to seven times lower than in the Standard method and up to 32 times lower than CFU). The CFU method underestimated the concentration of viable conidia in most of the tested fungal forms, and in the emulsifiable oil products, these values were 54% lower for B. bassiana and 84% lower for M. anisopliae. The adaptations and standardizations proposed in the ESALQ method showed effective improvements for routine quality assessment of

  10. The Adh1 gene of the fungus Metarhizium anisopliae is expressed during insect colonization and required for full virulence.

    Science.gov (United States)

    Callejas-Negrete, Olga Alicia; Torres-Guzmán, Juan Carlos; Padilla-Guerrero, Israel Enrique; Esquivel-Naranjo, Ulises; Padilla-Ballesteros, Maria Fernanda; García-Tapia, Adriana; Schrank, Augusto; Salazar-Solís, Eduardo; Gutiérrez-Corona, Félix; González-Hernández, Gloria Angélica

    2015-03-01

    Zymography of alcohol dehydrogenase (ADH) activity in the entomopathogenic fungus Metarhizium anisopliae grown under various conditions revealed that micro-aerobic growth was associated with increased ADH activity. The major ADH protein, AdhIp, was purified to homogeneity by affinity chromatography and has an estimated molecular weight of 41kDa and an isoelectric point (pI) of 6.4. Peptide mass fingerprint analysis allowed the identification and cloning of the gene that encodes this protein, Adh1, as annotated in the M. anisopliae genome database. AdhIp is related to the medium-chain dehydrogenase/reductase (MDR)/zinc-dependent alcohol dehydrogenase-like family and contains conserved ADH sequence motifs, such as the zinc-containing ADH signature, the FAD/NAD binding domain and amino acid residues that are conserved in most microbial ADHs. Semi-quantitative RT-PCR analysis revealed that Adh1 gene expression occurs at low levels during early Plutella xylostella infection and that the Adh1 gene was primarily expressed at larval death and as mycelia emerge from the insect cuticle before conidiation. Antisense-RNA experiments indicated that NAD(+)-dependent ADH activity was diminished by 20-75% in the transformants, and the transformants that had lower ADH activity showed allyl alcohol resistance, which indicates that reduction in ADH activity also occurs in vivo. Bioassays performed using antisense adh1 transformants, which have lower ADH activity, showed that LC50 values were two to five times higher than the wild-type, indicating that AdhIp is required for full capability of the fungus to penetrate and/or colonize the insect.

  11. Horizontal Transmission of Metarhizium anisopliae in Fruit Flies and Effect of Fungal Infection on Egg Laying and Fertility

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    Nguya K. Maniania

    2013-05-01

    Full Text Available Fly-to-fly transmission of conidia of the entomopathogenic fungus Metarhizium anisopliae and the effect of fungal infection on the reproductive potential of females surviving infection were investigated in three fruit fly species, Ceratitis cosyra, C. fasciventris, and C. capitata. The number of conidia picked up by a single fruit fly was determined in C. cosyra. The initial uptake (Day 0 of conidia by a single fly was approx. 1.1 × 106 conidia after exposure to the treated substrate. However, the number of conidia dropped from 7.2 × 105 to 4.1 × 105 conidia after 2 and 8 h post-exposure, respectively. The number of conidia picked up by a single fungus-treated fly (“donor” varied between 3.8 × 105 and 1.0 × 106 in the three fruit fly species, resulting in 100% mortality 5–6 days post-exposure. When fungus-free flies of both sexes (“recipient” flies were allowed to mate with “donor” flies, the number of conidia picked up by a single fly varied between 1.0 × 105 and 2.5 × 105, resulting in a mortality of 83–100% in C. capitata, 72–85% in C. cosyra and 71–93% in C. fasciventris 10–15 days post-inoculation. There was an effect of fungal infection on female egg laying in the three species of fruit flies as control flies laid more eggs than fungus-treated females. The percentage reduction in fecundity in flies infected with M. anisopliae was 82, 73 and 37% in C. capitata, C. fasciventris and C. cosyra, respectively. The results are discussed with regard to application in autodissemination techniques.

  12. The physiological effects of multi-walled carbon nanotubes (MWCNTs) on conidia and the development of the entomopathogenic fungus, Metarhizium anisopliae (Metsch.) Sorok.

    Science.gov (United States)

    Gorczyca, Anna; Kasprowicz, Marek J; Lemek, Tadeusz

    2014-01-01

    The aim of the study was an in vitro evaluation of the effect of MWCNTs on the conidia of two strains of entomopathogenic fungus, Metarhizium anisopliae. The study made use of water suspensions of MWCNTs (concentration ∼ 3 mg·mL(-1)) made from commercial nanotubes and centrifuged. The conidia were placed in contact with nanotubes for 240 h. An assessment of MWCNT influence on conidia was performed after 1, 24, 72 and 240 h and focused on the linear growth of vegetative mycelium derived from these conidia, mycelium sporulation in subcultures and pathogenicity. Using TEM imaging, it was demonstrated that carbon nanotubes are able to damage cell membranes of the examined fungi conidia. However, the absence was noted of a significantly fungistatic effect of both MWCNT suspensions on the examined strains with respect to the physiological features in question. The increase in vegetative mycelium effected by spores after contact with MWCNTs was characterized by a slight modification in relation to the control. There was no strong trend (inhibition - stimulation), in relation to the effect of the tested suspension of carbon nanotubes, on the development of the vegetative mycelium in in vitro culture. Sporulation of the mycelium after completion of the culture only occurred in one case (strain Ma73F and culture of spores after 24-h contact with MWCNTs) significantly more intensely than in the controls. With respect to pathogenicity for test insects compared to the control strain, Ma73F spores grown from the longest contact with nanotubes suspensions performed significantly better. On the basis of the calculated of mycelium index growth rates and the time of death of the test insects (LT50), it was found that the adverse effects of water suspension MWCNTs on the spores of M. anisopliae were applied after a short contact with biological material. This indicates unfavorable physical rather than chemical effects on the tested cell. Over time, nanotube aggregation in water

  13. Imbibitional damage in conidia of the entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, and Metarhizium acridum

    Science.gov (United States)

    When dried organisms are immersed in water, rapid imbibition may cause severe damage to plasma membranes; in unicellular organisms, such damage is usually lethal. This study investigated effects of pre-immersion moisture levels and immersion temperature on imbibitional damage in three insect pathoge...

  14. Copulation Activity, Sperm Production and Conidia Transfer in Aedes aegypti Males Contaminated by Metarhizium anisopliae: A Biological Control Prospect.

    Science.gov (United States)

    Garza-Hernández, Javier A; Reyes-Villanueva, Filiberto; Russell, Tanya L; Braks, Marieta A H; Garcia-Munguia, Alberto M; Rodríguez-Pérez, Mario A

    2015-01-01

    Dengue is the most prevalent arboviral disease transmitted by Aedes aegypti worldwide, whose chemical control is difficult, expensive, and of inconsistent efficacy. Releases of Metarhizium anisopliae--exposed Ae. aegypti males to disseminate conidia among female mosquitoes by mating represents a promising biological control approach against this important vector. A better understanding of fungus virulence and impact on reproductive parameters of Ae. aegypti, is need before testing auto-dissemination strategies. Mortality, mating competitiveness, sperm production, and the capacity to auto-disseminate the fungus to females up to the 5 th copulation, were compared between Aedes aegypti males exposed to 5.96 x 10(7) conidia per cm2 of M. anisopliae and uninfected males. Half (50%) of fungus-exposed males (FEMs) died within the first 4 days post-exposure (PE). FEMs required 34% more time to successively copulate with 5 females (165 ± 3 minutes) than uninfected males (109 ± 3 minutes). Additionally, fungus infection reduced the sperm production by 87% at 5 days PE. Some beneficial impacts were observed, FEMs were able to successfully compete with uninfected males in cages, inseminating an equivalent number of females (about 25%). Under semi-field conditions, the ability of FEMs to search for and inseminate females was also equivalent to uninfected males (both inseminating about 40% females); but for the remaining females that were not inseminated, evidence of tarsal contact (transfer of fluorescent dust) was significantly greater in FEMs compared to controls. The estimated conidia load of a female exposed on the 5th copulation was 5,200 mL(-1) which was sufficient to cause mortality. Our study is the first to demonstrate auto-dissemination of M. anisopliae through transfer of fungus from males to female Ae. aegypti during mating under semi-field conditions. Our results suggest that auto-dissemination studies using releases of FEMs inside households could successfully

  15. Copulation Activity, Sperm Production and Conidia Transfer in Aedes aegypti Males Contaminated by Metarhizium anisopliae: A Biological Control Prospect.

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    Javier A Garza-Hernández

    Full Text Available Dengue is the most prevalent arboviral disease transmitted by Aedes aegypti worldwide, whose chemical control is difficult, expensive, and of inconsistent efficacy. Releases of Metarhizium anisopliae--exposed Ae. aegypti males to disseminate conidia among female mosquitoes by mating represents a promising biological control approach against this important vector. A better understanding of fungus virulence and impact on reproductive parameters of Ae. aegypti, is need before testing auto-dissemination strategies.Mortality, mating competitiveness, sperm production, and the capacity to auto-disseminate the fungus to females up to the 5 th copulation, were compared between Aedes aegypti males exposed to 5.96 x 10(7 conidia per cm2 of M. anisopliae and uninfected males. Half (50% of fungus-exposed males (FEMs died within the first 4 days post-exposure (PE. FEMs required 34% more time to successively copulate with 5 females (165 ± 3 minutes than uninfected males (109 ± 3 minutes. Additionally, fungus infection reduced the sperm production by 87% at 5 days PE. Some beneficial impacts were observed, FEMs were able to successfully compete with uninfected males in cages, inseminating an equivalent number of females (about 25%. Under semi-field conditions, the ability of FEMs to search for and inseminate females was also equivalent to uninfected males (both inseminating about 40% females; but for the remaining females that were not inseminated, evidence of tarsal contact (transfer of fluorescent dust was significantly greater in FEMs compared to controls. The estimated conidia load of a female exposed on the 5th copulation was 5,200 mL(-1 which was sufficient to cause mortality.Our study is the first to demonstrate auto-dissemination of M. anisopliae through transfer of fungus from males to female Ae. aegypti during mating under semi-field conditions. Our results suggest that auto-dissemination studies using releases of FEMs inside households could

  16. Seleção de isolados de Metarhizium anisopliae e Beauveria bassiana patogênicos a soldados de Atta bisphaerica e Atta sexdens rubropilosa em condições de laboratório Selection of Metarhizium anisopliae and Beauveria bassiana isolates pathogenic to Atta bisphaerica and Atta sexdens rubropilosa soldiers under laboratory conditions

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    Alzimiro Marcelo Conteiro Castilho

    2010-06-01

    crops, pastures and planted forests. The entomopathogenic fungi are among the natural mortality factors of these ants and because of that they have potential to be use in biological control of this pest. The present research aimed to select isolates of Metarhizium anisopliae and Beauveria bassiana pathogenic to soldiers of Atta sexdens rubropilosa and Atta bisphaerica under laboratory conditions. To evaluate the pathogenicity, eight isolates of M. anisopliae and six of B. bassiana were used. The experiment was carried out in a completely randomized design, being the plots composed by a group of ten soldiers per each ant species, and three replicates per treatment. For each isolate, three groups of soldiers were sprayed with conidial suspensions containing 1.0 x 10(8 conidia ml-1, kept in moist chamber (25±1°C, 80 1% of RH in the dark without food, and the mortality was evaluated every day. From the 14 isolates tested, four of M. anisopliae and four of B. bassiana were pathogenic to soldiers of the both ant species. The virulence was evaluated to the isolates that caused mortality equal or higher than 50%. For each isolate, suspensions containing 1.0 x 10(6 to 1.0 x 10(11 conidia ml-1 were sprayed on three groups of ten soldiers, and equally kept as on the pathogenicity test. The mortality percentage was calculated each 24 hours for determining TL50. The isolate ENA04 of M. anisopliae was the most pathogenic, causing more than 80% of mortality in the first three days after inoculation, showed higher capacity of spore production on ant cadavers, and was the most virulent to the soldiers of A. bisphaerica, with a TL50 of 1.15 days. All pathogenic isolates of A. sexdens rubropilosa soldiers were equally virulent.

  17. 绿僵菌毒素的研究进展%Research Progress of Metarhizium anisopliae Destruxin

    Institute of Scientific and Technical Information of China (English)

    张秋

    2009-01-01

    With highly toxic infectivity acting on pest ,Metarhizium anisopliae destruxin has a wide application prospect in the control of agricultural pests. It can be divided into low-molecular-weight compound and high-molecular-weight protein destruxin according to its molecular weight and configuration. The article particularly expatiated its molecular structure,category,extractingMethod,biology activity and producing condition.%绿僵菌毒素是绿僵菌产生的具有杀虫活性的次生代谢物质,在农业害虫防治方面具有广阔的应用前景.根据其分子量和结构分为低分子量毒素和高分子量蛋白毒素两大类,对其分子结构、种类、提取方法、生物活性、产生条件等进行了综述.

  18. Efficacy of water- and oil-in-water-formulated Metarhizium anisopliae in Rhipicephalus sanguineus eggs and eclosing larvae.

    Science.gov (United States)

    Luz, Christian; D'Alessandro, Walmirton Bezerra; Rodrigues, Juscelino; Fernandes, Éverton Kort Kamp

    2016-01-01

    Conidia of the entomopathogenic fungus Metarhizium anisopliae (Ascomycota: Clavicipitaceae) were assessed against Rhipicephalus sanguineus (Arachnida: Ixodidae) eggs under laboratory conditions. Clusters of 25 eggs were applied either directly with the fungal conidial formulations or set on previously fungus-treated filter paper. Treatments consisted of conidia formulated in water or an oil-in-water emulsion at final concentrations of 3.3 × 10(3), 10(4), 3.3 × 10(4), 10(5), or 3.3 × 10(5) conidia/cm(2). The development of mycelium and new conidia on egg clusters incubated at 25 °C and humidity close to saturation depended on conidial concentration, formulation, and application technique. No larvae eclosed from eggs after direct applications of conidia regardless of the formulation. The eclosion and survival of larvae from indirectly treated egg clusters depended on the type of formulation and conidial concentration applied. Oil-in-water formulations of conidia demonstrated the highest activity against eggs of R. sanguineus.

  19. Pathogenicity of Metarhizium anisopliae (Metch Sorok and Beauveria bassiana (Bals Vuill to adult Phlebotomus duboscqi (Neveu-Lemaire in the laboratory

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    Philip M. Ngumbi, Lucy W. Irungu, Paul N. Ndegwa & Nguya K. Maniania

    2011-03-01

    Full Text Available Background & objectives: Biological control of sandflies using entomopathogenic fungi is a possible alternativeto the expensive synthetic chemical control. It is potentially sustainable, less hazardous, and relatively inexpensiveand merits further investigations. The objective of this study was to identify the most pathogenic fungal isolate(sto sandflies in the laboratory.Methods: Isolates of entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana were screenedfor their pathogenicity against Phlebotomus duboscqi. Adult flies were contaminated using the technique describedby Migiro et al (2010. Briefly, flies were exposed to 0.1 g of dry conidia evenly spread on a cotton velvet clothcovering the inner side of a cylindrical plastic tube (95 mm long × 48 mm diam. In all 25 sandflies weretransferred into the cylindrical tube and allowed to walk on the velvet for one minute, after which they weretransferred from the velvet into the cages in Perplex. Insects in the control treatments were exposed to fungusfree velvet cloth before being transferred into similar cages. The treatments were maintained at 25 ± 2°C,60–70% RH and 12L: 12D photoperiod. The experiment was replicated 5 times. The most pathogenic isolateswere selected for further studies.Results: A total of 19 isolates were screened against adult sandflies in the laboratory. Mortality in the controlswas approximately 16.8 ± 1.7 %. All the isolates were found to be pathogenic to P. duboscqi. Mortality rangedbetween 76.8 and 100% on all the fungal isolates tested. The lethal time taken to 50% (LT50 and 90% (LT90(mortality ranged from 3.0–7.8 days and from 5.3–16.2 days, respectively. The virulent isolates, causing mortalitiesof 97.5–100%, were selected for further studies.Interpretation & conclusion: The high susceptibility of sandflies to entomopathogenic fungi suggests that fungiare potential alternatives to chemical control methods. We conclude that application of

  20. Efeito de Beauveria bassiana (Bals. Vuill. e Metarhizium anisopliae (Metsch. Sorok. sobre características biológicas de Diatraea saccharalis F. (Lepidoptera: Crambidae - DOI: 10.4025/actascibiolsci.v30i2.3627 Effect of Beauveria bassiana (Bals. Vuill. and Metarhizium anisopliae (Metsch. Sorok. on Biological Characteristics of Diatraea saccharalis F. (Lepidoptera: Crambidae - DOI: 10.4025/actascibiolsci.v30i2.3627

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    Valéria Wanderley Teixeira

    2008-05-01

    Full Text Available A broca da cana-de-açúcar Diatraea saccharalis F. é considerada uma das principais pragas nas Américas. Entre os métodos de controle, o uso de fungos entomopatogênicos tem sido amplamente recomendado no manejo das pragas da cana-de-açúcar, incluindo outras lepidobrocas. Assim sendo, este estudo investigou os efeitos de diferentes concentrações de Beauveria bassiana (Bals. Vuill. e Metarhizium anisopliae (Metsch. Sorok. sobre parâmetros biológicos da broca da cana-de-açúcar. Larvas de terceiro instar de D. saccharalis foram tratadas com os fungos usando as concentrações de 103, 104 e 105 conídios mL-1. Larvas tratadas com 105 conídios mL-1 de B. bassiana tiveram menor sobrevivência (56,6%, comparadas com lagartas não-tratadas (90%. Adultos originados de larvas tratadas colocaram menor número de ovos, com menor viabilidade, e viveram menos, comparados com adultos originados de larvas não-tratadas. Larvas tratadas com M. anisopliae na concentração de 105 conídios mL-1 e adultos originados destas larvas também exibiram redução no desempenho, comparados aos insetos não-tratados. Os resultados indicam que B. bassiana e M. anisopliae, além de patogênicos às larvas de D. saccharalis, também interferem negativamente na sua biologia, mostrando potencial de uso contra esta praga.The sugarcane borer Diatraea saccharalis F. is considered one of the major sugarcane pests in the American continent. Among control methods, the use of entomopathogenic fungi has been broadly recommended to manage sugarcane pests, including other sugarcane borers. Therefore, this study investigated the effects of different concentrations of Beauveria bassiana (Bals. Vuill and Metarhizium anisopliae (Metch. Sorok on biological characteristics of the sugarcane borer. Third-instar larvae of D. saccharalis were fungi-treated using the concentrations of 103, 104 and 105 conidia mL-1. Larvae treated with 105 conidia mL-1 of B. bassiana showed lower

  1. Potential of Metarhizium anisopliae (Metsch. Sorokin (Ascomycetes, hypocreales in the control of Bonagota salubricola (Meyrick (Lepidoptera, Tortricidae and its compatibility with chemical insecticides

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    FA. Anhalt

    Full Text Available Several insects are harmful to apples grown in Brazil, especially the leafroller Bonagota salubricola (Lepidoptera, Tortricidae, usually controlled with chemical insecticides. The purpose of the present study was to investigate the use of Metarhizium anisopliae strains in the control of the apple leafroller, by assessing their virulence to B. salubricola larvae in bioassays with suspensions of 2 x 10(5 to 2 x 10(9 conidia/mL as well as their relationship with protease expression. The most effective strain underwent a compatibility test with chemical insecticides. The M. anisopliae E6 strain showed a good performance, with up to 88% mortality and a LT50 of 1.66 days. The virulence was positively correlated with a higher enzymatic activity. The E6 strain was compatible with tebufenozide, evidencing its potential to control B. salubricola.

  2. Flutuação Populacional e Parasitismo de Ovos de Spodoptera frugiperda e Helicoverpa zea em milho pulverizado com extratos vegetais e Metarhizium anisopliae

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    Jaqueline Ramos de Freitas

    2010-10-01

    Full Text Available We evaluated the effect of plant extracts and Metarhizium anisopliae on population and parasitism of eggs of Spodoptera frugiperda and Helicoverpa zea in corn planted at two spacings. The research was conducted at the Agricultural School "Assis Chateaubriand" in Laguna Seca, PB, and Laborat6rio of Entomology at the Center for Integrated Pest Management / UEPB in Campina Grande-PB. Corn was planted in an area of 500m2, in spacing: (I 0,8 m x 0,4 m, and (II 0,4 m x 0,4 m, with one or two plants per hole, respectively. The treatments consisted of: 1 water as a control, 2 M. anisopliae (100 g 10 L-1 water, 3 Extract of Momordica charantia (50 ml L-1 water, 4 M. anisopliae (10 g L-1 water + water extract of M. charantia (50 ml L-1 water, 5 M. anisopliae (10 g L-1 water + cassava (Manihot esculenta Crantz (50 ml L-1 water. According to the results, no significant difference between treatments to control spacing I S. frugiperda egg, which was due, probably, late spraying, when 100% of the plants were attacked. The treatments also not efficient to control H zea eggs, recommending further studies to determine the concentration and dosage most appropriate for controlling these pests.

  3. Fitness costs to Helicoverpa armigera after exposure to sub-lethal concentrations of Metarhizium anisopliae sensu lato: Study on F1 generation.

    Science.gov (United States)

    Jarrahi, Azadeh; Safavi, Seyed Ali

    2016-07-01

    The entomopathogenic fungus, Metarhizium anisopliae (Metsch.) Sorokin is a valuable biocontrol agent attacking larval stages of many lepidopteran pests including Helicoverpa armigera (Hübner). Sub-lethal effects of M. anisopliae sensu lato (s.l.) (isolate M14) were investigated on life table parameters of offspring from treated larvae of H. armigera. Duration of different life stages was significantly affected by fungal treatments. Fecundity was decreased in females derived from H. armigera larvae treated with M. anisopliae s.l. Sub-lethal concentrations of the entomopathogen reduced the net reproduction rate (R0) of F1 insects for all treatments compared with the control. Similar reductions were observed for the intrinsic and the finite rates of increase (rm and λ, respectively). The mean generation time (T) and the doubling time (DT) were statistically higher in offspring of individuals exposed to some fungal concentrations than control insects. Our results indicated that there was a significant decrease in the F1 population of H. armigera derived from larvae that were exposed to sub-lethal concentrations of M. anisopliae s.l.

  4. Inhibition of ecto-phosphatase activity in conidia reduces adhesion and virulence of Metarhizium anisopliae on the host insect Dysdercus peruvianus.

    Science.gov (United States)

    Cosentino-Gomes, Daniela; Rocco-Machado, Nathália; Santi, Lucélia; Broetto, Leonardo; Vainstein, Marilene H; Meyer-Fernandes, José Roberto; Schrank, Augusto; Beys-da-Silva, Walter O

    2013-05-01

    Metarhizium anisopliae is an entomopathogenic fungus with the ability to infect a broad range of arthropods, and have evolved distinct strategies for their attachment to hosts. Here, we describe the characterisation of ecto-phosphatase activity on the conidia surface of M. anisopliae and its relevance in the host interaction process. Ecto-phosphatase activity was linear for 60 min and during this time, was linear with the increase of cell density. The optimum pH was in the acidic range and some divalent metals, such as Cu(2+), Cd(2+) and Zn(2+), inhibited ecto-phosphatase activity. The activity was also reduced by phosphatase inhibitors. Importantly, the inhibition of phosphatase activity in conidia reduced the adhesion to Dysdercus peruvianus (Hemiptera: Pyrrhocoridae) integument and, consequently and indirectly, M. anisopliae infection. The results herein presented show, for the first time, the importance of ecto-phosphatase activity in M. anisopliae conidia and provide the first evidence of its direct involvement in adhesion and host infection.

  5. Pathogenicity of Metarrhizium anisopliae (Metsch.) Sorokin and Beauveria bassiana (Bals.) Vuillemin isolates to Scaptocoris carvalhoi Becker (Hemiptera, Cydnidae); Patogenicidade de isolados de Metarhizium anisopliae (Metsch.) Sorokin e de Beauveria bassiana (Bals.) Vuillemin a Scaptocoris carvalhoi Becker (Hemiptera, Cydnidae)

    Energy Technology Data Exchange (ETDEWEB)

    Xavier, Luciane Modenez Saldivar [Universidade Federal da Grande Dourados (UFGD), Dourados, MS (Brazil)], e-mail: luciane_modenez@ibest.com.br; Avila, Crebio Jose [EMBRAPA Agropecuaria Oeste, Dourados, MS (Brazil)], e-mail: crebio@cpao.embrapa.br

    2006-12-15

    Pathogenicity of the fungi Metarrhizium anisopliae (Metsch.) Sorokin and Beauveria bassiana (Bals.) Vuillemin to stink bug Scaptocoris carvalhoi Becker, 1967 was evaluated under laboratory and greenhouse conditions. Experiments were carried out at EMBRAPA Agropecuaria Oeste, Dourados, Mato Grosso do Sul State, Brazil, in 2003. Ten M. anisopliae and eleven B. bassiana isolates were evaluated in laboratory using a completely randomized experimental design with five replicates (10 adults and 5 nymphs/plot). The pathogenicity of M. anisopliae isolate (Ma69) was also separately evaluated against nymphs and adults in laboratory and greenhouse. The stink bug mortality levels were higher for M. anisopliae isolates (between 73.3% and 94.7% than for B. bassiana isolates (between 10.7% and 78.7%). In greenhouse, stink bug mortality due to the M. anisopliae isolate (Ma69) was 57.3%, and there was no difference of mortality for nymphs and adults of stink bug in laboratory. However, in greenhouse, mortality levels were significantly higher (p<0,05) for nymphs (38,4%) than for adults (16,2%). From these data, we conclude that M. anisopliae isolate Ma69 was efficient to control S. carvalhoi in laboratory and in greenhouse, thus being a promising choice for use as a microbial insecticide under field conditions. (author)

  6. Efficacy of spray applications of entomopathogenic fungi against western flower thrips infesting greenhouse impatiens under variable moisture conditions

    Science.gov (United States)

    Efficacy tests of three entomopathogenic fungi (Beauveria bassiana strain GHA, Metarhizium brunneum strain F52, and Metarhizium anisopliae s.l. strain ESC-1) were conducted against thrips infesting greenhouse crops of single impatiens under variable moisture conditions. Fungal conidia suspended in 0...

  7. Eventos externos e internos da infecção de larvas e ninfas de Rhipicephalus sanguineus por Metarhizium anisopliae External and internal events of Rhipicephalus sanguineus larvae and nymphs infection by Metarhizium anisopliae

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    M.V. Garcia

    2008-08-01

    Full Text Available Examinaram-se a adesão, a germinação, a penetração e a colonização de larvas e ninfas de Rhipicephalus sanguineus por Metarhizium anisopliae, assim como as lesões infringidas pelo fungo nas respectivas fases do ciclo de vida do ácaro. Realizaram-se infecções experimentais em 11 grupos contendo 250 larvas e 11 grupos contendo 75 ninfas de R. sanguineus, por meio de banho, durante três minutos sob agitação manual, em suspensão contendo 10(8 conídios/ml do fungo. Nos grupos-controles, o banho foi realizado usando o veículo da suspensão. Larvas e ninfas foram processadas para um estudo histopatológico e de microscopia eletrônica de varredura nos seguintes tempos após a infecção: uma e 18 horas, e um, dois, três, quatro, cinco, seis, sete, nove e 11 dias. A germinação dos conídios ocorreu em até 18 horas pós-inoculação, e o fungo penetrou nas larvas e ninfas através do tegumento, dois e três dias após a infecção, respectivamente. Após penetração, o fungo invadiu o corpo das larvas e ninfas, promovendo uma colonização difusa, sem preferência aparente por tecidos específicos. Lesões significativas não foram observadas. A morte das larvas e ninfas ocorreu no terceiro e quarto dias pós-infecção, e a esporulação do patógeno sobre o cadáver foi iniciada no sexto dia pós-infecção.The adhesion, germination and colonization of Rhipicephalus sanguineus larvae and nymphs by Metarhizium anisopliae as well as the lesions caused by the fungus were studied. For this purpose, 11 groups of 250 larvae each and 11 groups of 75 nymphs each were bathed during 3 minutes under manual shaking in a 10(8 conidia/ml suspension. Corresponding control groups were bathed only in the suspension vehicle. Ticks were also submitted to both conventional microscopy and scanning eletronmicrocopy analyses at several post-infection periods (1 and 18 hours and 1, 2, 3, 4, 5, 6, 7, 9, and 11 days. Conidial germination occurred in less

  8. População de Metarhizium anisopliae em diferentes tipos e graus de compactação do solo Metarhizium anisopliae population in differents soil types and compactness degrees

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    Lourenço Maurício Lanza

    2004-12-01

    Full Text Available Este trabalho objetivou investigar a influência do tipo e compactação do solo na sobrevivência do fungo Metarhizium anisopliae. A sobrevivência do fungo foi determinada em quatro tipos de solos: Latossolo Vermelho textura argilosa, Latossolo Vermelho textura média, Argissolo Vermelho Amarelo textura arenosa média e Argissolo Vermelho Amarelo textura areno-argilosa, com maior teor de matéria orgânica. Para determinar o efeito da compactação na sobrevivência do fungo usaram-se os três primeiros tipos de solos nas densidades de 1,12, 1,32, 1,50g cm-3; 1,22, 1,44, 1,65g cm-3; 1,30, 1,50, 1,70g cm-3, respectivamente. Por meio da contagem de unidades formadoras de colônias (UFC em placas de Petri, fizeram-se avaliações da sobrevivência do fungo, após zero, 20, 40, 60, 80, 100 e 120 dias de incubação a 27 ± 1ºC. Houve influência significativa do tipo de solo e do grau de compactação na sobrevivência do fungo, obtendo-se maior quantidade de UFC no solo textura areno-argilosa. Entre os demais solos, a maior sobrevivência ocorreu no solo textura arenosa e a menor no solo textura argilosa. O efeito da compactação foi significativo para o tipo de solo, exceto no solo textura arenosa. Independentemente do tipo de solo, a maior sobrevivência foi observada nos valores médios de densidade. A compactação teve maior impacto no solo textura média, onde ocorreu queda mais acentuada na quantidade de UFC em todas as densidades.The aim of this work was to investigate the effect of different soil types and compactness on Metarhizium anisopliae survival. The fungus survivorship was determined in four soil types: Red Latosol with clay texture, Red Latosol with medium texture, Red Yellow Podzolic with medium sand texture and Red Yellow Podzolic with sand-clay texture, with higher amount of organic material. To determine the effect of compactness on fungus survival the first three soil types were used with the following densities

  9. Effects of conidial densities and spray volume of Metarhizium anisopliae and Beauveria bassiana fungal suspensions on conidial viability, droplet size and deposition coverage in bioassay using a novel bioassay spray system

    Science.gov (United States)

    Experiments were conducted to study the conidial viability during bioassay spray with different suspensions of Metarhizium anisopliae ATCC 62176 and Beauveria bassiana NI8, and to investigate the effects of conidial density and spray volume on the distribution of droplet size and deposit coverage us...

  10. [Isolation and identification of Metarhizium from Citrus grandis 'tomentosa' GAP base].

    Science.gov (United States)

    Ma, Weisi; Xu, Jiang; Qiao, Haili; Chen, Jun; Li, Xiangming; Qin, Rongmin; Cheng, Huizhen

    2012-04-01

    A type of entomopathogenic fungus of soil in Citrus grandis 'tomentosa' production base was isolated and identified with morphological and molecular biological methods, including pathogenesis, spore characteristic and ITS sequence analysis were conducted. The results showed that eighteen entomopathogenic fungi strains were isolated from the Tenebrio molitor infected in the soil samples, which were identified as Metarhizium anisopliae var. anisopliae. Based on results above, we concluded that there was quantity of Metarhizium resources in this area. These provided the useful information for controlling some pests of C. grandis by using these strains of fungus.

  11. Laboratory evaluation of the entomopathogenic fungus, Metarhizium anisopliae for the control of the groundnut bruchid, Caryedon serratus on groundnut.

    Science.gov (United States)

    Ekesi, S; Egwurube, E A.; Akpa, A D.; Onu, I

    2001-10-01

    The pathogenicity of five isolates of Metarhizium anisopliae to adult Caryedon serratus was evaluated in the laboratory. All the isolates tested were virulent to the beetle but pathogenicity varied among the isolates. One isolate, CPD 4 was consistently superior to all other isolates in terms of mortality of the beetle, protection of groundnut pods from damage, reduction in progeny production and repellency to the beetle. At 10 days post-treatment, adult mortality treated with 0.1, 0.5 and 1.0g of dry conidia equivalent to 3.6x10(8), 1.8x10(9) and 3.6x10(9) conidia of isolate CPD 4 per 50g of groundnut pods was 100% which did not differ significantly from pirimiphos-methyl-treated pods at 10ppm. At the lowest dosage of 0.1g of conidia per 50g of pods, damage in pods protected with isolate CPD 4 was 5% which did not differ significantly from the 2% damage in pods protected by pirimiphos-methyl at 10ppm but significantly differed from damage in untreated pods which was 26%. Isolate CPD 4 caused complete reduction in progeny emergence at all dosages tested. It also exhibited some degree of repellency to the beetle with percentage repellency values of between 40-79% at concentrations of 0.1-1.0g of conidia per 50g of groundnut pods. These combined virulence and repellency characteristics of this isolate may increase its protectant potential against C. serratus.

  12. Interactions between Entomopathogenic Fungus, Metarhizium Anisopliae and Sublethal Doses of Spinosad for Control of House Fly, Musca Domestica

    Directory of Open Access Journals (Sweden)

    M Sharififard

    2011-06-01

    Full Text Available Background: Metarhizium anisopliae strain IRAN 437C is one of the most virulent fungal isolates against house fly, Musca domestica. The objective of this study was to determine the interaction of this isolate with sublethal doses of spino­sad against housefly.Methods: In adult bioassay, conidia of entomopathogenic fungus were applied as inoculated bait at 105 and 107 spore per gram and spinosad at 0.5, 1 and 1.5 µg (A.I. per gram bait. In larval bioassay, conidia were applied as combina­tion of spore with larval bedding at 106 and 108 spore per gram and spinosad at sublethals of 0.002, 0.004 and 0.006 µg (AI per gram medium. Results: Adult mortality was 48% and 72% for fungus alone but ranged from 66–87% and 89–95% in combination treat­ments of 105 and 107 spore/g with sublethal doses of spinosad respectively. The interaction between 105 spore/g with sublethals exhibited synergistic effect, but in combination of 107 spore in spite of higher mortality, the interac­tion was additive. There was significant difference in LT50 among various treatments. LT50 values in all combination treat­ments were smaller than LT50 values in alone ones. Larval mortality was 36% and 69% for fungus alone but ranged from 58%–78% and 81%–100% in combination treatments of 106 and 108 spore/g medium with sublethals of spino­sad respectively. The interaction was synergistic in all combination treatments of larvae.Conclusion: The interaction between M. anispliae and spinosad indicated a synergetic effect that increased the house fly mortality as well as reduced the lethal time.

  13. Isolation of Metarhizium anisopliae carboxypeptidase A with native disulfide bonds from the cytosol of Escherichia coli BL21(DE3).

    Science.gov (United States)

    Austin, Brian P; Waugh, David S

    2012-03-01

    The carboxypeptidase A enzyme from Metarhizium anisopliae (MeCPA) has broader specificity than the mammalian A-type carboxypeptidases, making it a more useful reagent for the removal of short affinity tags and disordered residues from the C-termini of recombinant proteins. When secreted from baculovirus-infected insect cells, the yield of pure MeCPA was 0.25mg per liter of conditioned medium. Here, we describe a procedure for the production of MeCPA in the cytosol of Escherichia coli that yields approximately 0.5mg of pure enzyme per liter of cell culture. The bacterial system is much easier to scale up and far less expensive than the insect cell system. The expression strategy entails maintaining the proMeCPA zymogen in a soluble state by fusing it to the C-terminus of maltose-binding protein (MBP) while simultaneously overproducing the protein disulfide isomerase DsbC in the cytosol from a separate plasmid. Unexpectedly, we found that the yield of active and properly oxidized MeCPA was highest when coexpressed with DsbC in BL21(DE3) cells that do not also contain mutations in the trxB and gor genes. Moreover, the formation of active MeCPA was only partially dependent on the disulfide-isomerase activity of DsbC. Intriguingly, we observed that most of the active MeCPA was generated after cell lysis and amylose affinity purification of the MBP-proMeCPA fusion protein, during the time that the partially purified protein was held overnight at 4°C prior to activation with thermolysin. Following removal of the MBP-propeptide by thermolysin digestion, active MeCPA (with a C-terminal polyhistidine tag) was purified to homogeneity by immobilized metal affinity chromatography (IMAC), ion exchange chromatography and gel filtration.

  14. Infection of malaria (Anopheles gambiae s.s.) and filariasis (Culex quinquefasciatus) vectors with the entomopathogenic fungus Metarhizium anisopliae.

    Science.gov (United States)

    Scholte, Ernst-Jan; Njiru, Basilio N; Smallegange, Renate C; Takken, Willem; Knols, Bart G J

    2003-09-15

    Current intra-domiciliary vector control depends on the application of residual insecticides and/or repellents. Although biological control agents have been developed against aquatic mosquito stages, none are available for adults. Following successful use of an entomopathogenic fungus against tsetse flies (Diptera: Glossinidae) we investigated the potency of this fungus as a biological control agent for adult malaria and filariasis vector mosquitoes. In the laboratory, both sexes of Anopheles gambiae sensu stricto and Culex quinquefasciatus were passively contaminated with dry conidia of Metarhizium anisopliae. Pathogenicity of this fungus for An. gambiae was further tested for varying exposure times and different doses of oil-formulated conidia. Comparison of Gompertz survival curves and LT50 values for treated and untreated specimens showed that, for both species, infected mosquitoes died significantly earlier (p control groups. No differences in LT50 values were found for different exposure times (24, 48 hrs or continuous exposure) of An. gambiae to dry conidia. Exposure to oil-formulated conidia (doses ranging from 1.6 x 10(7) to 1.6 x 10(10) conidia/m2) gave LT50 values of 9.69 +/- 1.24 (lowest dose) to 5.89 +/- 0.35 days (highest dose), with infection percentages ranging from 4.4-83.7%. Our study marks the first to use an entomopathogenic fungus against adult Afrotropical disease vectors. Given its high pathogenicity for both adult Anopheles and Culex mosquitoes we recommend development of novel targeted indoor application methods for the control of endophagic host-seeking females.

  15. Infection of malaria (Anopheles gambiae s.s. and filariasis (Culex quinquefasciatus vectors with the entomopathogenic fungus Metarhizium anisopliae

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    Smallegange Renate C

    2003-09-01

    Full Text Available Abstract Background Current intra-domiciliary vector control depends on the application of residual insecticides and/or repellents. Although biological control agents have been developed against aquatic mosquito stages, none are available for adults. Following successful use of an entomopathogenic fungus against tsetse flies (Diptera: Glossinidae we investigated the potency of this fungus as a biological control agent for adult malaria and filariasis vector mosquitoes. Methods In the laboratory, both sexes of Anopheles gambiae sensu stricto and Culex quinquefasciatus were passively contaminated with dry conidia of Metarhizium anisopliae. Pathogenicity of this fungus for An. gambiae was further tested for varying exposure times and different doses of oil-formulated conidia. Results Comparison of Gompertz survival curves and LT50 values for treated and untreated specimens showed that, for both species, infected mosquitoes died significantly earlier (p 50 values were found for different exposure times (24, 48 hrs or continuous exposure of An. gambiae to dry conidia. Exposure to oil-formulated conidia (doses ranging from 1.6 × 107 to 1.6 × 1010 conidia/m2 gave LT50 values of 9.69 ± 1.24 (lowest dose to 5.89 ± 0.35 days (highest dose, with infection percentages ranging from 4.4–83.7%. Conclusion Our study marks the first to use an entomopathogenic fungus against adult Afrotropical disease vectors. Given its high pathogenicity for both adult Anopheles and Culex mosquitoes we recommend development of novel targeted indoor application methods for the control of endophagic host-seeking females.

  16. Construction of transgenic Trichoderma koningi with chit42 of Metarhizium anisopliae and analysis of its activity against the Asian corn borer.

    Science.gov (United States)

    Li, Ying Y; Tang, Jun; Fu, Ke H; Gao, Shi G; Wu, Qiong; Chen, Jie

    2012-01-01

    The chit42 gene was cloned from Metarhizium anisopliae CY1 and was inserted into Trichoderma koningii T30 genome by protoplast transformation. Sixteen transgenic isolates were identified by polymerase chain reaction analysis. The chit42 gene was 1275 bp in length and its coded protein was approximately 42 kDa in size. Semi-quantitative RT-PCR analysis and the measurement of the chitinase activity under induced conditions were conducted. Mortality of the Asian corn borer (Ostrinia furnacalis) was used for assessing efficacy of culture filtrates and conidial suspensions of transgenic Trichoderma strains against the insect. The results indicated that the transgenic Trichoderma strain harboring chit42 gene from Metarhizium anisopliae CY1 showed significant lethal effect on the Asian corn borer larvae. Study on growth inhibition of silkworm (Bombyx mori) larvae was carried out. The transgenic Trichoderma could better hinder the growth and development of the silkworm larvae than the wild-type Trichoderma did. The inhibition to the expression of three genes associated with development and anti-stress response in the mid-gut of the Asian corn borer larvae was more significant in the transcriptional level after larvae were fed with transgenic biomass than with the wild type. Evaluation of inhibition on the growth of maize ear rot pathogens was carried out in vitro test and the transgenic strains kept antagonistic activity against Fusarium verticilloides.

  17. Comparative impact of artificial selection for fungicide resistance on Beauveria bassiana and Metarhizium brunncum

    Science.gov (United States)

    Hypocreales fungi such as Beauveria bassiana and Metarhizium anisopliae can be negatively affected by fungicides thereby reducing their biocontrol potential. The overall goal of this study was to investigate the impact of artificial selection for fungicide resistance on two commercial entomopathoge...

  18. Production of microsclerotia by brazilian strains of metarhizium spp. using submerged liquid culture fermentation

    Science.gov (United States)

    We investigated the potential production and desiccation tolerance of microsclerotia (MS) by Brazilian strains of Metarhizium. anisopliae [Ma], M. acridum [Mc] and M. robertsii [Mr]. These fungi were grown in a liquid medium containing 16 g carbon l-1 with a carbon:nitrogen ratio of 50:1. One hundre...

  19. Inhititory Mechanisms of Metarhizium anisopliae Against Rhizoctonia cerealis%绿僵菌对小麦纹枯病菌的抑制作用研究

    Institute of Scientific and Technical Information of China (English)

    齐永霞; 陈方新; 李增智

    2011-01-01

    The inhibition and mechanism of Metarhizium anisopliae against Rhizoctonia cerealis was studied in vitro by applying dual-culture, cup-plate test and microscopy, respectively. The results showed that in dual-culture tests and cupplate tests, the tested M. Anisopliae had strong antagonism against the tested R. Cerealis. It suggested that the inhibitory effects of the tested M. Anisopliae on the tested R. Cerealis were significant when it was dual-cultured and when its conidia suspension was applied into the PDA media. Using metabolic liquid of Ma55, which was cultured in different time, the inhibitions to hyphal growth, sclerotium-formation number and sclerotium weight, sclerotium-germinating rate were studied, too. The results showed that the metabolic liquid cultured 25 d has significant inhibitory effect on hyphal growth, sclerotium number and sclerotium-germinating rate. The test indicated that the culture filtrates of Ma55 contained a thermal stable antifungal compound. It was also observed that the hyphal of Metarhizium anisopliae and Rhizoctonia cerealis has no hyper parasitism, but the hyphal of Rhizoctonia cerealis was lysed, broken and their cytoplasm becamethinning on dual culture. The results mentioned above indicated that the inhibitory effects of Metarhizium anisopliae against the pathogen of wheat sharp eyespot were produced mostly by nutrition competition, space competition and antibiosis.%在实验室条件下,研究了金龟子绿僵菌(Metarhizium anisopliae)对小麦纹枯病菌(Rhizoctonia cerealis)的拮抗作用及其机理.结果表明,金龟子绿僵菌与小麦纹枯病菌对峙培养以及在培养基中加入金龟子绿僵菌孢子悬浮液,对小麦纹枯病菌菌丝生长均有较好的抑制作用.测定了培养不同天数的金龟子绿僵菌Ma55发酵液对小麦纹枯病菌菌丝生长、菌核产生量及菌核萌发率的影响.结果表明,液体振荡培养25 d的金龟子绿僵菌Ma55发酵液对小麦纹枯病菌的菌丝

  20. Secretome of the biocontrol agent metarhizium anisopliae induced by the cuticle of the cotton pest Dysdercus peruvianus reveals new insights into infection.

    Science.gov (United States)

    Beys-da-Silva, Walter O; Santi, Lucélia; Berger, Markus; Calzolari, Diego; Passos, Dario O; Guimarães, Jorge A; Moresco, James J; Yates, John R

    2014-05-02

    Metarhizium anisopliae is an entomopathogenic fungus that has evolved specialized strategies to infect insect hosts. Here we analyzed secreted proteins related to Dysdercus peruvianus infection. Using shotgun proteomics, abundance changes in 71 proteins were identified after exposure to host cuticle. Among these proteins were classical fungal effectors secreted by pathogens to degrade physical barriers and alter host physiology. These include lipolytic enzymes, Pr1A, B, C, I, and J proteases, ROS-related proteins, oxidorreductases, and signaling proteins. Protein interaction networks were generated postulating interesting candidates for further studies, including Pr1C, based on possible functional interactions. On the basis of these results, we propose that M. anisopliae is degrading host components and actively secreting proteins to manage the physiology of the host. Interestingly, the secretion of these factors occurs in the absence of a host response. The findings presented here are an important step in understanding the host-pathogen interaction and developing more efficient biocontrol of D. peruvianus by M. anisopliae.

  1. Sublethal effect of concentrations of Metarhizium anisopliae (Metsch.) Sorok on the larval stage and immunologic characteristics of Diatraea flavipennella (BOX) (Lepidoptera: Crambidae).

    Science.gov (United States)

    Guimarães, Jennifer; Marques, Edmilson J; Wanderley-Teixeira, Valéria; De Albuquerque, Auristela C; Dos Passos, Eliana M; Silva, Cínthia C M; Teixeira, Álvaro A C

    2014-12-01

    This work aimed to evaluate the effects of Metarhizium anisopliae on Diatraea flavipennella and investigate their immune response. Was observed the effect of M. anisopliae against larvae of D. flavipennella sprayed at concentrations of 103, 104, 105 conidia / mL, in which showed differences relative the larval period, extending up to 72.0 days in treatment and 25.0 days in the control. The results for hemocytes revealed that the most frequent cells when sprayed at the concentrations of 103, 105, 107 conidia / mL were the prohemocytes, spherulocytes, plasmatocytes and granulocytes in relation to adipohemocytes and oenocytoids. The level of nitric oxide was different between the control and the concentration 107 spores / mL (24), while the activity of phenoloxidase was similar among treatments in 24 and higher concentration 107 spores / mL (60h). In biochemical profile of hemocytes was a change in carbohydrates, lipids and proteins in response to the fungus. The results indicate that the fungus M. anisopliae can be used in the Integrated Management of D. flavipennella by presenting pathogenicity and interfere with their development even when exposed to small concentrations.

  2. Effect of Metarhizium anisopliae on the fertility and fecundity of two species of fruit flies and horizontal transmission of mycotic infection.

    Science.gov (United States)

    Sookar, P; Bhagwant, S; Allymamod, M N

    2014-01-01

    In Mauritius, the peach fruit fly, Bactrocera zonata Saunders (Diptera: Tephritidae), and the melon fly, Bactrocera cucurbitae (Coquillett), are the major pest of fruits and vegetables, respectively. Fruit growers make use of broad-spectrum insecticides to protect their crops from fruit fly attack. This method of fruit fly control is hazardous to the environment and is a threat to beneficial insects. The entomopathogenic fungus, Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae), which was isolated from the soils of Mauritius, was used to investigate whether fungus-treated adult fruit flies could transfer conidia to non-treated flies during mating, and whether fungal infection could have an effect on mating behavior, fecundity, and fertility of the two female fruit fly species. When treated male flies were maintained together with non-treated female flies, they were able to transmit infection to untreated females, resulting in high mortalities. Similarly, fungus-infected female flies mixed with untreated males also transmitted infections to males, also resulting in high mortalities. Infection by M. anisopliae also resulted in the reduction of the number of eggs produced by females of B. cucurbitae. The results suggest that M. anisopliae may have potential for use in integrated control programs of B. zonata and B. cucurbitae using the sterile insect technique in Mauritius.

  3. Sublethal effect of concentrations of Metarhizium anisopliae (metsch. sorok on the larval stage and immunologic characteristics of Diatraea flavipennella (box (lepidoptera: crambidae

    Directory of Open Access Journals (Sweden)

    JENNIFER GUIMARÃES

    2014-12-01

    Full Text Available This work aimed to evaluate the effects of Metarhizium anisopliae on Diatraea flavipennella and investigate their immune response. Was observed the effect of M. anisopliae against larvae of D. flavipennella sprayed at concentrations of 103, 104, 105 conidia / mL, in which showed differences relative the larval period, extending up to 72.0 days in treatment and 25.0 days in the control. The results for hemocytes revealed that the most frequent cells when sprayed at the concentrations of 103, 105, 107 conidia / mL were the prohemocytes, spherulocytes, plasmatocytes and granulocytes in relation to adipohemocytes and oenocytoids. The level of nitric oxide was different between the control and the concentration 107 spores / mL (24, while the activity of phenoloxidase was similar among treatments in 24 and higher concentration 107 spores / mL (60h. In biochemical profile of hemocytes was a change in carbohydrates, lipids and proteins in response to the fungus. The results indicate that the fungus M. anisopliae can be used in the Integrated Management of D. flavipennella by presenting pathogenicity and interfere with their development even when exposed to small concentrations.

  4. Evaluation of Metarhizium anisopliae var. anisopliae Qu-M845 Isolate to Control Varroa destructor (Acari: Varroidae in Laboratory and Field Trials Evaluación del Aislamiento Qu-M845 de Metarhizium anisopliae var. anisopliae para el Control de Varroa destructor (Acari: Varroidae en Ensayos de Laboratorio y Terreno

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    Marta Rodríguez

    2009-12-01

    Full Text Available The effectiveness of the Metarhizium anisopliae (Metschinkoff Qu-M845 isolate was evaluated in laboratory and field trials. It was previously selected for thermal resistance (at 30 and 35 ºC and pathogenicity on Varroa destructor (Anderson and Trueman. In the laboratory, the first evaluations were carried out by spraying and increasing the concentration from zero to 10(8 conidia mL-1 on varroa adults. Lethal concentrations required for 50 and 90% mite mortality (LC50 and LC90 were 3.8 x 10(5 and 8 x 10(7 conidia mL-1, respectively (χ2 = 2.03. In the autumn field trials, three application methods (doses of 5 x 10(10 conidia per hive were evaluated. The treatments were: a conidia stamped on filter paper, located on every second frame inside the hive; b dry conidia sprinkled on and between frames; and c dry conidia in a dispenser path at the entrance of the hive. Furthermore, untreated hives were included as controls. After 21 days of treatment, the dry conidia sprinkled on and between frames showed 67% less bees infested by the mite than the control (p La efectividad del aislamiento Qu-M845 de Metarhizium anisopliae (Metschinkoff, seleccionado previamente por su resistencia a temperaturas de 30 y 35 ºC, y patogenicidad sobre Varroa destructor Anderson y Trueman fue evaluada en laboratorio y en ensayos de terreno. Las primeras pruebas consistieron en pulverizar concentraciones crecientes de 0 a 10(8 conidias mL-1 sobre varroas adultas. La concentración letal para matar el 50 y 90% de la población (CL50 y CL90 fueron de 3,8 x 10(5 y 8 x 10(7 conidias mL-1, respectivamente (χ² = 2,03. En otoño se evaluaron en terreno tres métodos de aplicación de una dosis de 5 x 10(10 conidias por colmena. Los tratamientos fueron: a conidias estampadas en papel filtro ubicado cada dos panales móviles al interior de la colmena; b conidias espolvoreadas sobre y entre los panales; y c dispensador de conidias ubicado en la piquera de las colmenas. Además se

  5. Histopathological events and detection of Metarhizium anisopliae using specific primers in infected immature stages of the fruit fly Anastrepha fraterculus (Wiedemann, 1830) (Diptera: Tephritidae).

    Science.gov (United States)

    Bechara, I J; Destéfano, R H R; Bresil, C; Messias, C L

    2011-02-01

    The fungus Metarhizium anisopliae is used on a large scale in Brazil as a microbial control agent against the sugar cane spittlebugs, Mahanarva posticata and M. fimbriolata (Hemiptera., Cercopidae). We applied strain E9 of M. anisopliae in a bioassay on soil, with field doses of conidia to determine if it can cause infection, disease and mortality in immature stages of Anastrepha fraterculus, the South American fruit fly. All the events were studied histologically and at the molecular level during the disease cycle, using a novel histological technique, light green staining, associated with light microscopy, and by PCR, using a specific DNA primer developed for M. anisopliae capable to identify Brazilian strains like E9. The entire infection cycle, which starts by conidial adhesion to the cuticle of the host, followed by germination with or without the formation of an appressorium, penetration through the cuticle and colonisation, with development of a dimorphic phase, hyphal bodies in the hemocoel, and death of the host, lasted 96 hours under the bioassay conditions, similar to what occurs under field conditions. During the disease cycle, the propagules of the entomopathogenic fungus were detected by identifying DNA with the specific primer ITSMet: 5' TCTGAATTTTTTATAAGTAT 3' with ITS4 (5' TCCTCCGCTTATTGATATGC 3') as a reverse primer. This simple methodology permits in situ studies of the infective process, contributing to our understanding of the host-pathogen relationship and allowing monitoring of the efficacy and survival of this entomopathogenic fungus in large-scale applications in the field. It also facilitates monitoring the environmental impact of M. anisopliae on non-target insects.

  6. The Control Effect of Metarhizium anisopliae and Regent to Grasshopper%应用绿僵菌与锐劲特防治蝗虫的效果

    Institute of Scientific and Technical Information of China (English)

    董辉; 高松; 农向群; 丛斌; 张泽华

    2011-01-01

    The control test to grasshopper was conducted with oil-based suspension and bait of Metarhizium anisopliae and Regent. Investigation was taken by the methods of sample and sweeping. Results showed that the grasshopper began to die in the third day after oil-based suspension and bait of M. Anisopliae were applied. The mortality increased significantly in the 15th day, and the mortality exceeded 80% in the 50th day. The oil-based suspension of M. Anisopliae had better control effect on grasshopper than bait of M. Anisopliae. The insecticidal rate of the oil-based suspension of Regent was significantly higher than of the M. Anisopliae, but the persistence was significantly lower. The insecticidal rate and persistence of Regent bait were intermediate. In conclusion, the oil-based suspension of M. Anisopliae could be used as the preferred biological a-gents against grasshopper.%利用绿僵菌与锐劲特制备的油悬浮剂和饵剂在内蒙古草原进行了防治蝗虫试验.结果表明,利用样筐调查和扫网调查发现在施用绿僵菌油悬浮剂和饵剂后第三天蝗虫开始感病死亡,第15天死亡率显著增加,第50天死亡率超过80%,绿僵菌油悬浮剂具有比绿僵菌饵剂更好的防治效果;锐劲特油悬浮剂的杀虫速率显著高于绿僵菌,但持效性显著低于绿僵菌;锐劲特饵剂的杀虫速率和持效性居中.绿僵菌油悬浮剂可以作为防治草原蝗虫的首选生物制剂.

  7. Colonização e lesão em fêmeas ingurgitadas do carrapato Rhipicephalus sanguineus causadas pelo fungo Metarhizium anisopliae Colonization and lesions on engorged female Rhipicephalus sanguineus, caused by Metarhizium anisopliae

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    Marcos Valerio Garcia

    2004-10-01

    Full Text Available O presente trabalho teve como objetivo verificar a forma de penetração do fungo Metarhizium anisopliae [METSCH. (SOROKIN, 1883] em carrapatos da espécie Rhipicephalus sanguineus (LATREILLE, 1806, assim como as lesões infringidas nos tecidos internos do ácaro. A forma de aderência e penetração do fungo foi estudada através da microscopia eletrônica de varredura e a ação do fungo nos tecidos internos avaliada em secções histológicas convencionais. Para observação destes eventos, realizaram-se infecções experimentais em 11 grupos de fêmeas ingurgitadas do carrapato R. sanguineus contendo 12 fêmeas ingurgitadas cada. Para tal, as fêmeas ingurgitadas foram banhadas durante 3 minutos, sob agitação manual, em suspensão com concentração 108 conídios/mL. No caso dos grupos controle o banho foi realizado apenas no veículo da suspensão. Os carrapatos foram processados para histopatologia e microscopia eletrônica em diversos tempos após a infecção, a saber: 1 e 18h, e um, dois, três, quatro, cinco, seis, sete, nove e onze dias. Observou-se que a maior parte dos conídios germinou em até 18h após a inoculação e que o fungo penetrou no ácaro através do tegumento 48h após a infecção. Após a penetração, o fungo invadiu o corpo do hospedeiro promovendo uma colonização difusa, sem preferência aparente por tecidos específicos. Dentre as lesões nos tecidos internos do ácaro, ressalta-se o rompimento da parede intestinal e vazamento do conteúdo para a hemocele. A morte do hospedeiro ocorreu entre 96 e 120h pós-infecção, e a esporulação do patógeno sobre o cadáver do ácaro iniciou-se em torno de 120 a 144h pós-infecção. Espera-se, com este trabalho, contribuir para o desenvolvimento e viabilização de técnicas de controle biológico dos carrapatos por fungos como alternativa ao uso de acaricidas.The objective of this work was to verify the penetration of the fungus Metarhizium anisopliae [METSCH

  8. Eficiência de Metarhizium anisopliae no controle do Percevejo-do-Colmo Tibraca limbativentris (Heteroptera: Pentatomidae em lavoura de arroz irrigado Efficiency of Metarhizium anisopliae on rice stem bug Tibraca limbativentris (Heteroptera: Pentatomidae control in flooded rice field

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    José Francisco da Silva Martins

    2004-12-01

    Full Text Available O percevejo-do-colmo, Tibraca limbativentris Stal, 1860 (Heteroptera: Pentatomidae, é altamente prejudicial à cultura do arroz no Brasil, principalmente em sistemas de cultivo irrigados por inundação. O efeito de duas formas de aplicação, conídios em suspensão e veiculados em grãos de arroz autoclavado, da cepa (CP 172 de Metarhizium anisopliae (Metsch. Sorok., no controle do percevejo, foi avaliado em três experimentos conduzidos em 1991, 1993 e 1994, em lavoura comercial de arroz irrigado. Em 1991, tanto a pulverização de conídios como a distribuição manual grãos de arroz cobertos com material fúngico em dosagem equivalente a 7,2 x 10(13 conídios.ha-1, sobre o solo e entre os colmos de arroz, onde os percevejos estavam alojados, reduziram significativamente a população natural do inseto com eficiência de controle de 52,6% e 61,8%, respectivamente. Ainda em 1991, estudo sobre o estabelecimento e persistência da cepa no solo do arrozal, com base na contagem de unidades formadoras de colônias (UFC, indicou que o fungo manteve-se viável na entressafra, até 216 dias após a aplicação, época da implantação de novos arrozais. O número de UFC foi maior nas parcelas tratadas com os grãos de arroz cobertos com material fúngico. O crescimento linear do número de UFC, no solo nas parcelas testemunhas, evidenciou disseminação do fungo para partes anteriormente não tratadas do arrozal. Resultados significativos de controle com a aplicação da suspensão de conídios e dos grãos de arroz com o fungo, na dose de 5 x 10(13 conídios.ha-1, também foram obtidos em 1993, com 51,8% e 48,2% de eficiência, respectivamente. Em 1994, a aplicaç��o da suspensão de 5 x 10(13 conídios.ha-1 resultou em 39,5% de controle do inseto. O índice de confirmação de infeção, contudo, foi baixo, nos dois anos, atingindo, no máximo, 20%, em 1993. O baixo número de insetos com micose, em comparação ao índice de mortalidade

  9. 绿僵菌野生菌株对红脉穗螟幼虫的致病效果研究%Bioassay of Toxicity of Metarhizium anisopliae Wild Strains to Larvae of Tirathaba Rufivena

    Institute of Scientific and Technical Information of China (English)

    吕朝军; 钟宝珠; 钱军; 苟志辉; 孙晓东; 覃伟权

    2014-01-01

    采用孢子悬浮液喷雾法测定不同绿僵菌( Metarhizium anisopliae)野生菌株对红脉穗螟( Tirathaba rufivena)幼虫的致病能力,旨在获得对该虫具有高致病力的生防菌株,以期为红脉穗螟的生物防治提供新的生物资源。结果表明,G2和G3菌株对红脉穗螟幼虫表现出较高致病力,在1.0×108孢子/mL浓度下,两个菌株对红脉穗螟幼虫的累计校正死亡率分别为86.32%和80.34%,LT50分别为4.77 d、6.18 d;田间试验表明,G2和G3菌株在1.0×1010孢子/mL浓度下,药后7 d可使红脉穗螟的虫口减退率分别达到73.72%和68.71%,表明这两株菌株在红脉穗螟生物防治中是极具有应用潜力的生防真菌。%In order to screen entomopathogenic fungi strains with high virulence against the larva of Ti⁃rathaba rufivena,the evaluation of the pathogenicity of Metarhizium anisopliae to T.rufivena larvae was conduc⁃ted using the spore suspension spray method. The results indicated that trains of G2 and G3 had higher viru⁃lence to T.rufivena larvae,and the cumulative mortality rates were 86.32% and 80.34%,respectively;the me⁃dian lethal times (LT50) were 4.77 d and 6.18 d at the concentration of 1.0×108 spores·mL-1.The field trial results showed that the pest population declined by 73.72% and 68.71% after treated with G2 and G3 at the concentration of 1.0×108 spores·mL-1 7 d after the treatment.The results suggest that G2 and G3 M.anisopliae strains have potential in biocontrol of T.rufivena.

  10. Laboratory Trials of Metarhizium anisopliae var. acridum (Green Muscle® Against the Saxaul Locust, Dericorys albidula Serville (Orthoptera: Dericorythidae Ensayos de Laboratorio de Metarhizium anisopliae var. acridum (Green muscle® contra la Langosta de Saxaul, Dericorys albidula Serville (Orthoptera: Dericorythidae

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    Heydar Valizadeh

    2011-12-01

    Full Text Available The saxaul locust, Dericorys albidula Serville (Orthoptera: Dericorythidae is a major pest of saxaul plants in Qom province of Iran. During 2005-2006, different nymphal instars of bands of D. albidula were treated by aerial spraying of Metarhizium anisopliae var. acridum (Green Muscle®. The gasoline formulation of M. anisopliae var. acridum isolate IMI 330189 was applied in different conidial concentrations (10(6, 10(7, 10(8, 10(9, 10(10 and 10(13 spores mL-1 that were prepared in sterile distilled gasoline. Results showed that various concentrations significantly affected the 2nd, 3rd, 4th and 5th nymphal instars of D. albidula compared to control. In addition, there were no differences in the effects of the different concentrations in 2005, but the differences were significant in 2006. Concentration 10(10 killed 100% of tested insects 15 d after treatment. Comparing the results of the two years showed that the susceptibility of nymphs in the second year (2006 was higher than in the first year (2005. In conclusion, the results of this study indicated that the fungal insecticide M. anisopliae var. acridum, diluted in gasoline, was efficacious with the nymphal instars of locust D. albidula in 2005 and 2006.La langosta del saxaul, Dericorys albidula Serville (Orthoptera: Dericorythidae, es la principal plaga de plantas de saxaul de las provincias Qom, Irán. Durante 2005-2006, diferentes instars ninfales de D. albidula se asperjaron con Metarhizium anisopliae var. acridum (Green Muscle®. La formulación de gasolina de M. anisopliae var. acridum aislamiento IMI 330189 se aplicó en diferentes concentraciones conidiales (10(6, 10(7, 10(8, 10(9, 10(10 and 10(13 esporas mL-1 preparadas en gasolina destilada estéril. Los resultados demostraron que varias concentraciones afectaron significativamente los nstars ninfales de D. albidula comparado con el control. Además el efecto de concentraciones no fue diferente en 2005 pero sí significativamente en

  11. Seletividade de agrotóxicos utilizados na cultura do arroz irrigado ao fungo Metarhizium anisopliae, agente de controle microbiano de Tibraca limbativentris Selectivity of chemical pesticides used in rice irrigated crop at fungus Metarhizium anisopliae, microbial control agent of Tibraca limbativentris

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    Fátima Teresinha Rampelotti-Ferreira

    2010-04-01

    Full Text Available Para subsidiar a utilização de táticas de manejo integrado para o inseto-praga Tibraca limbativentris, avaliou-se a compatibilidade in vitro entre o isolado CG 891 de Metarhizium anisopliae e os agrotóxicos utilizados na cultura do arroz irrigado. Os parâmetros avaliados foram crescimento vegetativo, esporulação e germinação em pré e pós-contato do fungo, com duas doses de fipronil, tiametoxam, fenitrotiona, carbofurano, pirazossulfurom-etílico, bentazona, clomazona, glifosato e quincloraque; e uma dose de triciclazol e azoxistrobina. Reduções significativas no crescimento vegetativo e esporulação de M. anisopliae foram observadas para fenitrotiona, carbofurano e azoxistrobina. Observou-se redução na germinação em pré-contato para o agrotóxico azoxistrobina. O valor percentual da germinação alterou a classificação dos agrotóxicos com relação a sua seletividade sobre o entomopatógeno. No entanto, os resultados mostram que in vitro os agrotóxicos fenitrotiona, carbofurano, glifosato e azoxistrobina são prejudiciais ao isolado CG 891 de M. anisopliae.Searching for integrated control strategies for the insect-pest Tibraca limbativentris, the in vitro compatibility between isolated CG 891 of Metarhizium anisopliae and several pesticides used in the culture of irrigated rice was evaluated. The followings parameters were studied: vegetative growth, sporulation and the germination viability in pre- and post-contact of the fungus for two concentrations of fipronil, thiametoxan, fenitrothion, carbofuran, pyrazosulfuron-ethyl, bentazon, clomazone, glyfosate and quinclorac and one concentration of tricyclazole and azoxystrobin. Significant reductions in the vegetative growth and sporulation of M. anisopliae were observed for fenitrothion, carbofuran and azoxystrobin. The germination in pre-contact was affected by azoxystrobin. The percentage of germination changed the classification of pesticides in relation to its

  12. Occurrence of entomopathogenic fungi in arable soil

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    Ryszard Miętkiewski

    2014-08-01

    Full Text Available Samples of soil were taken from arable field and from balk. Larvae of Galleria mellonella and Ephestia kühniella were used as an "insect bait" for isolation of entomopathogenic fungi from soil. Metarhizium anisopliae and Paecilomyces fumosoroseus were isolated from both kind of soil. but Beauveria bassiana was present only in soil taken from balk.

  13. AVALIAÇÃO DA VIRULENCIA DE BLASTOSPOROS DE Metarhizium anisopliae NO CONTROLE DE LARVAS DE CAMPO DO MOSQUITO Aedes aegypti

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    Aline Teixeira Carolino

    2016-11-01

    Full Text Available Atualmente no Brasil, o mosquito Aedes aegypti é transmissor das arboviroses dengue, chikungunya e Zika. Não existe tratamento específico para estas doenças. A redução da população do vetor ainda é o método mais eficaz para reduzir a taxa dessas arboviroses. O presente estudo comparou a virulência de conídios e blastosporos de Metarhizium anisopliae contra larvas do mosquito A. aegypti provenientes de coletas no campo. Blastosporos foram mais virulentos para larvas, sendo observada mortalidade total das larvas em apenas 48 horas. Larvas infectadas com conídios apresentaram 100% de mortalidade no quinto dia pós-infecção. O presente estudo mostra que blastosporos apresentam grande potencial para controle de larvas de A. aegypti no campo.

  14. Metarhizium anisopliae and Its Mechanism for Killing Insects%金龟子绿僵菌及其杀虫机理

    Institute of Scientific and Technical Information of China (English)

    裘晖; 吴振强; 梁世中

    2004-01-01

    金龟子绿僵菌(Metarhizium anisopliae)属于半知菌亚门绿僵菌属,是一种昆虫内寄生菌物.它对寄主侵染过程包括粘附、孢子萌发、穿透虫体、体内发育和致死.这一过程是附着胞、表皮降解酶和破坏菌素等物质的生理生化作用的综合结果.由于其寄主范围广,致病力强,对人、畜、农作物无毒,无残毒、菌剂易生产,持效期长等优点,具有广阔的应用前景.

  15. Screening of Metarhizium anisopliae Strains for Controlling Brontispa longissima%感染椰心叶甲绿僵菌菌株的筛选

    Institute of Scientific and Technical Information of China (English)

    吴青; 曾玲; 徐大高; 陆永跃

    2006-01-01

    从田间感病的椰心叶甲虫尸上分离所得病原菌,经逐步筛选获得最优的绿僵菌Metarhizium anisopliae BMa-9菌株.其菌落生长率为6.14 mm/d,平均产分生孢子量3.91×109 mL-1,孢子萌发率87.32%.1×106 mL-1孢子悬浮液处理椰心叶甲时,对3龄幼虫具很强感染力,第3 d平均感染率61%,第8 d平均死亡率97%.

  16. 金龟子绿僵菌菌株生长环境变量的优化%Optimization of environmental factors for the growth of Metarhizium anisopliae var.anisopliae strains MA4 and Malml

    Institute of Scientific and Technical Information of China (English)

    杨腊英; 甘露; 刘丽; 文明富; 黄俊生

    2008-01-01

    金龟子绿僵菌(Metarhizium anisopliae var.anisopliae)是一种广谱的昆虫病原真菌,提高菌株的生长速率及产孢量具有重要的意义.分别对2株金龟子绿僵菌菌株MA4与MAlml的菌丝生长及次代产孢具有影响的培养温度、培养基初始pH、装液比、光照及微量元素等环境因素进行了测定.经过优化得到2菌株最佳培养条件分别为:MA4菌株在28℃、pH 7、装液比为75ml/250ml、加入微量元素Mn全光照培养时生长最好、次代产孢量最高;MAlml菌株在28℃、pH9、装液比为75ml/250ml、加入微量元素Cu、黑暗培养时生长最好、次代产孢量最高.

  17. Avaliação do potencial de controle biológico do Metarhizium anisopliae sobre Boophilus microplus em teste de estábulo Evaluation of the biological control potential of Metarhizium anisopliae toward Boophilus microplus in pen trials

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    Thiago C. Bahiense

    2007-12-01

    Full Text Available Avaliou-se por meio de um teste de estábulo a capacidade de Metarhizium anisopliae em controlar o carrapato Boophilus microplus. Foram realizados tratamentos em bovinos estabulados, utilizando este fungo aplicando-se suspensão conidial em banhos por aspersão. Os resultados foram obtidos pelo percentual de mortalidade de carrapatos durante 28 dias após o tratamento e pelos índices biológicos após incubação destes em câmara climatizada. Constatou-se 33% de mortalidade no período total observado, sendo o índice de produção de ovos e índice nutricional reduzidos somente em um curto período após o tratamento.The aim of the present study was to evaluate the capacity of the fungus Metarhizium anisopliae to control Boophilus microplus tick in pen trials. Infested calves were held in individual pen and treated with fungus suspension through aspersion bath. The results were evaluated based on ticks' mortality rate for 28 days after treatment, and on the analysis of biology of tick's samples which were transferred to an incubation chamber. It was reported 33% of mortality during the total period analyzed, and the production of eggs and nutritional rates were decreased only for a short period after treatment.

  18. O USO DE SOLUÇÃO PROTETORA PREVINE PERDA DE VIABILIDADE DE BLASTOSPOROS DE Metarhizium anisopliae DURANTE A LIOFILIZAÇÃO

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    Aline Teixeira Carolino

    2017-04-01

    Full Text Available Em cultura líquida, o fungo Metarhizium anisopliae é capaz de produzir estruturas denominadas blastosporos. Para utilização a seco, é necessária a secagem do fungo, e o método mais utilizado para tal processo é a liofilização. Entretanto, é sabido que este processo pode levar a perda de viabilidade de algumas culturas. O presente estudo investigou a possibilidade de utilização de uma solução protetora pré-liofilização para promover proteção a blastosporos de M. anisopliae. Após a liofilização, através de ensaios de unidade formadora de colônia (UFC, foi evidenciado que o fungo seco na solução contendo leite em pó + trealose + glutamato de sódio foi o mais capaz de manter a viabilidade e integridade celular do fungo durante o processo de secagem, comparada as demais soluções testadas. Sendo observada médias de 180 colônias do fungo seco com a solução protetora supracitada e 1 colônia do fungo seco apenas em água estéril.

  19. Enhancement of microbial hydroxylation of 13-ethyl-gon-4-ene-3,17-dione by Metarhizium anisopliae using nano-liposome technique.

    Science.gov (United States)

    Feng, Meiqing; Liao, Ziwei; Han, Lei; Li, Jiyang; Ye, Li

    2014-04-01

    The introduction of 11α-hydroxy to 13-ethyl-gon-4-ene-3,17-dione (GD) by microbial transformation is a key step in the synthesis of oral contraceptive desogestrel, while low substrate solubility and uptake into cells are tough problems influencing biotransformation efficiency greatly. Nano-liposome technique was used in the hydroxylation of GD by Metarhizium anisopliae. The substrate GD was processed to be GD-loaded nano-liposomes (GNLs) with high stability and encapsulation efficiency, and then applied in microbial hydroxylation by M. anisopliae. The results proved that the yield of the main product 11α-hydroxy-13-ethyl-gon-4-ene-3,17-dione (HGD) tripled compared to regular solvent dimethylformamide dispersion method at 2 g/l of substrate feeding concentration, and the HGD conversion rate showed no obvious reduction when the substrate feeding concentration increased from 2 to 6 g/l, which indicated the improvement of GNL addition method on biotransformation. Furthermore, the main byproduct changed from 6β-hydroxy derivative of GD (with similar polarity to HGD) to 6β,11α-dihydroxy derivative, which benefits the following purification of HGD from fermentation broth. These advantages suggest a great potential for the application of nano-liposome technique in microbial steroid transformation.

  20. Evaluation of cellulose substrates treated with Metarhizium anisopliae (Metschnikoff Sorokin as a biological control agent against the termite Microcerotermes diversus Silvestri (Isoptera: Termitidae

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    Behzad Habibpour

    2011-08-01

    Full Text Available This article is the first report on the promising effect of an entomopathogenic fungus, Metarhizium anisopliae (Metschnikoff Sorokin to control populations of Microcerotermes diversus Silvestri. Biological control is an alternative to the long-term usage of chemical pesticides. M. anisopliae, the causal agent of green muscardine disease of insects, is an important fungus in biological control of insect pests. Bait systems can eliminate entire colonies of subterranean termites. Baiting reduces adverse environmental impacts caused by organochlorine and organophosphate pesticides in the control of termites and creates sustainable protection of buildings against their invasion. Treated-sawdust bait was applied by two methods: a combination of treated sawdust and untreated filter paper, and b combination of treated sawdust and untreated sawdust. When combinations of treated sawdust and untreated sawdust were used, LC50 and LC90 were 8.4×106 and 3.9×107 (spore/ml, respectively. With the use of improved bait formula and more virulent strains, we hope to achieve better control of termite colonies and enable pathogens to become a useful element in the Integrated Pest Management system.

  1. Construction and preliminary analysis of a normalized cDNA library from Locusta migratoria manilensis topically infected with Metarhizium anisopliae var. acridum.

    Science.gov (United States)

    Wang, Jie; Xia, Yuxian

    2010-08-01

    The insect immune response to fungal infection is poorly understood at the molecular level. To explore the molecular basis of this process, a novel method to analyze the gene transcripts of insects in response to pathogenic fungus was established. A normalized cDNA library based on the SMART method combined with DSN (duplex-specific nuclease) treatment was constructed using mRNA extracted from the fat body and hemocytes of Locusta migratoria manilensis 6-24h after being topically infected with Metarhizium anisopliae var. acridum. Analysis of 259 unigenes out of 303 sequenced inserts from the cDNA library revealed that the cDNA library was not contaminated with M. anisopliae transcripts and validated the presence of the immune-related genes characterized here. These results suggest that this method overcame the difficulties of contamination from a fungal source in constructing the host cDNA library from mycosed insects and proved that this method is reliable and feasible for investigation of host genes in response to fungal infection. Further studies of the expressed sequence tags from this library will provide insights into the molecular basis of insect immune response to fungal infection.

  2. Identification of genes differentially expressed in vivo by Metarhizium anisopliae in the hemolymph of Locusta migratoria using suppression-subtractive hybridization.

    Science.gov (United States)

    Zhang, Chuanbo; Xia, Yuxian

    2009-08-01

    Metarhizium anisopliae is an important insect pathogenic fungus widely used in biological pest control. The aim of this study was to identify genes differentially expressed in vivo by M. anisopliae CQMa102 in the hemolymph of infected Locusta migratoria. Suppression-subtractive hybridization was performed using cDNA generated from hyphal bodies purified from hemolymph and the fungus germinating and differentiating on locust wings. A total of 350/1,600 random clones screened by cDNA array dot blotting were sequenced, resulting in 120 uniquely expressed sequence tags (ESTs) that were up-regulated during colonization of hemolymph. Among these 120 ESTs, 42 (35.0%) had matches in the NR protein database, and 29 (24.2%) were significantly similar to known proteins involved in various cellular processes, including general metabolism, cell wall remodeling, protein synthesis, signal transduction and stress responses. In contrast, the remaining 78 ESTs (65.0%) either had low similarity in the NR database or represented novel genes. Semi-quantitative RT-PCR analysis of five randomly selected genes revealed that all were highly expressed in the host hemolymph. These results provide new insight into the underlying molecular mechanisms of adaptation to host hemolymph and may increase understanding of host-pathogen interactions.

  3. On the Separation and Liquid Culture of Metarhizium anisopliae (Metsch) Sorokin (L8500)%金龟子绿僵菌(L8500)的分离及液体培养

    Institute of Scientific and Technical Information of China (English)

    郭素萍; 徐一强; 曹素芳

    2004-01-01

    通过液体发酵培养对定名为金龟子绿僵菌(Metarhizium anisopliae(Metsch)Sorokin)(L8500)的培养基进行了考察,确定了该菌液体培养的最佳配方,并且制定了L8500菌液体培养的质量控制方案.

  4. Purification of destruxins produced by Metarhizium anisopliae and bioassay of their insecticidal activities against grubs%绿僵菌素的分离制备及其对蛴螬的毒力

    Institute of Scientific and Technical Information of China (English)

    胡琼波; 任顺祥; 刘树艳

    2007-01-01

    以金龟子绿僵菌金龟子变种Metarhizium anisopliae var.anisopliae菌株MaQ10发酵液为材料,采用萃取、浓缩、制备色谱及重结晶技术,分离纯化出5种绿僵菌素的晶体,经与标准样品以及

  5. Effects of physical and nutritional stress conditions during mycelial growth on conidial germination speed, adhesion to host cuticle, and virulence of Metarhizium anisopliae, an entomopathogenic fungus.

    Science.gov (United States)

    Rangel, Drauzio E N; Alston, Diane G; Roberts, Donald W

    2008-11-01

    Growth under stress may influence pathogen virulence and other phenotypic traits. Conidia of the entomopathogenic fungus Metarhizium anisopliae var. anisopliae (isolate ARSEF 2575) were produced under different stress conditions and then examined for influences on in vitro conidial germination speed, adhesion to the insect cuticle, and virulence to an insect host, Tenebrio molitor. Conidia were produced under non-stress conditions [on potato-dextrose agar plus 1gl(-1) yeast extract (PDAY; control)], or under the following stress conditions: osmotic (PDAY+sodium chloride or potassium chloride, 0.6 or 0.8m); oxidative [(PDAY+hydrogen peroxide, 5mm) or UV-A (irradiation of mycelium on PDAY)]; heat shock (heat treatment of mycelium on PDAY at 45 degrees C, 40min); and nutritive [minimal medium (MM) with no carbon source, or on MM plus 3gl(-1) lactose (MML)]. Conidia were most virulent (based on mortality at 3d) and had the fastest germination rates when produced on MML, followed by MM. In addition, conidial adhesion to host cuticle was greatest when the conidia were produced on MML. Media with high osmolarity (0.8m) produced conidia with slightly elevated virulence and faster germination rates than conidia produced on the control medium (PDAY), but this trend did not hold for media with the lower osmolarity, (0.6m). Conidia produced from mycelium irradiated with UV-A while growing on PDAY had somewhat elevated virulence levels similar to that of conidia produced on MM, but their germination rate was not increased. Hydrogen peroxide and heat shock treatments did not alter virulence. These results demonstrate that the germination, adhesion and virulence of M. anisopliae conidia can be strongly influenced by culture conditions (including stresses) during production of the conidia.

  6. Control de Rhipicephalus microplus (Acari: Ixodidae mediante el uso del hongo entomopatógeno Metarhizium anisopliae (Hypocreales: Clavicipitaceae. Revisión

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    Melina Maribel Ojeda-Chi

    2011-01-01

    Full Text Available Las infestaciones de la garrapata del ganado, Rhipicephalus microplus, producen el mayor problema global de ectoparásitos en ganado de regiones tropicales y subtropicales, provocan importantes pérdidas económicas en la producción de carne, leche y pieles, además incrementan los gastos derivados de los programas de control, y son capaces de transmitir Babesia bovis, B. bigemina y Anaplasma marginale. El control de R. microplus se basa principalmente en el uso de ixodicidas, sin embargo, su uso irracional ha propiciado la aparición de garrapatas resistentes a las principales familias de ixodicidas, siendo necesario desarrollar alternativas de control no químico. Una de estas alternativas es el uso de hongos entomopatógenos, entre los que se encuentra Metarhizium anisopliae (Hypocreales, Clavicipitaceae el cual ha demostrado ser eficiente, tanto en estudios in vitro como in vivo, para el control de las diferentes fases evolutivas de R. microplus, causa disminución en la tasa de oviposición, incrementa el período de incubación y de eclosión, además produce la muerte de larvas y garrapatas adultas con porcentajes de eficiencia de hasta el 100 %. Diferentes estudios demuestran que M. anisopliae representa una alternativa no química sustentable para el control de garrapatas. La presente revisión tiene como objetivo presentar información actualizada sobre el uso de diferentes cepas de M. anisopliae en el control de la garrapata R.microplus.

  7. Efficacy of entomopathogenic hypocrealean fungi against Periplaneta americana.

    Science.gov (United States)

    Hubner-Campos, Rayssa Fátima; Leles, Renan Nunes; Rodrigues, Juscelino; Luz, Christian

    2013-12-01

    The American cockroach Periplaneta americana, one of the worlds' most important urban insect pests was tested with entomopathogenic fungi. Most promising Metarhizium anisopliae, Metarhizium robertsii and Beauveria bassiana killed nymphs (≥ 81.7% mortality, 25 days after treatment), and these fungi developed on all dead insects. Other fungi tested were less virulent (Metarhizium frigidum and Purpureocillium lilacinum) or avirulent (Isaria cateniobliqua, Isaria farinosa, Simplicillium lanosoniveum, Sporothrix insectorum and Tolypocladium cylindrosporum). Intrageneric and intraspecific variability of fungal activity was detected. Adults were highly susceptible, and oothecae proved to be more resistant than nymphs and adults to infection with M. anisopliae IP 46. Findings of the study underscore the potential of fungi as biocontrol agents against this pest.

  8. Kemampuan Larva Oryctes Rhinoceros (Coleoptera: Scarabaeidae) Menularkan Cendawan Metarhizium Anisopliae Ke Larva Sehat Di Pertanaman Kelapa Sawit

    OpenAIRE

    Perabu Jaya Sitepu

    2009-01-01

    Perabu Jaya Sitepu, Kemampuan larva Oryctes rhinoceros menularkan Metharizium anisopliae ke larva sehat di pertanaman kelapa sawit. Penelitian bertujuan untuk mengetahui kemampuan larva O. rhinoceros menularkan M. anisopliae ke larva O. rhinoceros yang lain di lapangan. Penelitian ini menggunakan Rancangan Acak Kelompok (RAK) Non Faktorial yang terdiri dari 4 perlakuan dan enam ulangan yaitu : L1 (kontrol), L2 (5 larva dilumuri M. anisoplioae), L3 (10 larva dilumuri M. anisoplioae), L4 (15 la...

  9. Production of destruxins from Metarhizium spp. fungi in artificial medium and in endophytically colonized cowpea plants.

    Directory of Open Access Journals (Sweden)

    Patrícia S Golo

    Full Text Available Destruxins (DTXs are cyclic depsipeptides produced by many Metarhizium isolates that have long been assumed to contribute to virulence of these entomopathogenic fungi. We evaluated the virulence of 20 Metarhizium isolates against insect larvae and measured the concentration of DTXs A, B, and E produced by these same isolates in submerged (shaken cultures. Eight of the isolates (ARSEF 324, 724, 760, 1448, 1882, 1883, 3479, and 3918 did not produce DTXs A, B, or E during the five days of submerged culture. DTXs were first detected in culture medium at 2-3 days in submerged culture. Galleria mellonella and Tenebrio molitor showed considerable variation in their susceptibility to the Metarhizium isolates. The concentration of DTXs produced in vitro did not correlate with percent or speed of insect kill. We established endophytic associations of M. robertsii and M. acridum isolates in Vigna unguiculata (cowpeas and Cucumis sativus (cucumber plants. DTXs were detected in cowpeas colonized by M. robertsii ARSEF 2575 12 days after fungal inoculation, but DTXs were not detected in cucumber. This is the first instance of DTXs detected in plants endophytically colonized by M. robertsii. This finding has implications for new approaches to fungus-based biological control of pest arthropods.

  10. Pathogenicity, Ovicidal Action, and Median Lethal Concentrations (LC 50 ) of Entomopathogenic Fungi against Exotic Spiralling Whitefly, Aleurodicus dispersus Russell

    OpenAIRE

    Boopathi Thangavel; Karuppuchamy Palaniappan; Kalyanasundaram Manickavasagam Pillai; Mohankumar Subbarayalu; Ravi Madhaiyan

    2013-01-01

    Biological control using entomopathogenic fungi could be a promising alternative to chemical control. Entomopathogenic fungi, Beauveria bassiana (Balsamo) Vuillemin, Metarhizium anisopliae (Metschnikoff) Sorokin, Lecanicillium lecanii (Zimmerm.) Zare and Gams, and Paecilomyces fumosoroseus (Wize) Brown and Smith, were tested for their pathogenicity, ovicidal effect, and median lethal concentrations (LC50) against exotic spiralling whitefly, Aleurodicus dispersus Russell. The applications were...

  11. Specific diversity of the entomopathogenic fungi Beauveria and Metarhizium in Mexican agricultural soils.

    Science.gov (United States)

    Pérez-González, Víctor H; Guzmán-Franco, Ariel W; Alatorre-Rosas, Raquel; Hernández-López, Jorge; Hernández-López, Antonio; Carrillo-Benítez, María G; Baverstock, Jason

    2014-06-01

    Prior knowledge of the local population structure of entomopathogenic fungi is considered an important requisite when developing microbial control strategies against major pests of crops such as white grubs. An extensive survey in the estate of Guanajuato, one of the main agricultural regions of Mexico, was carried out to determine the abundance and diversity of entomopathogenic fungi in soil. Soil collected from 11 locations was baited for entomopathogenic fungi using Galleria mellonella. In addition, all isolates were morphologically identified and selected isolates of Beauveria and Metarhizium isolates identified using Bloc and ITS or Elongation Factor 1-α and ITS sequence information respectively. Genotypic diversity was then studied using microsatellite genotyping. The proportion of isolates belonging to each genus varied amongst all locations. The species Beauveria bassiana, B. pseudobassiana and Metarhizium robertsii were found, with B. bassiana being the most abundant and widely distributed. Microsatellite genotyping showed that the 36 B. bassiana isolates were grouped in 29 unique haplotypes, but with no separation according to geographical origin.

  12. Entomopathogenic Fungi (Hypocreales) for Control of Potato Psyllid

    Science.gov (United States)

    Laboratory and field trials of the four isolates of entomopathogenic fungi, Isaria fumosorosea and Metarhizium anisopliae were conducted to evaluate their potential for control of the potato psyllid. In the laboratory, 2 ml aqueous suspension of 107conidia/ml in a of each isolate applied in a Potte...

  13. 雄甾-4-烯-3,17-二酮11α羟化突变株Metarhizium anisopliae M28-203的代谢调控%Metabolic regulation of androst-4-ene-3,17-dione 11α-hydroxylation mutant strain Metarhizium anisopliae M28-203

    Institute of Scientific and Technical Information of China (English)

    叶丽; 周珮; 冯美卿

    2011-01-01

    目的 从营养代谢调控和转化条件控制两个方面入手提高雄甾-4-烯-3,17-二酮(androst-4-ene-3,17-dione,AD)11α羟化突变株Metarhizium anisopliae M28-203转化效率.方法 通过正交试验确定发酵培养基配方,对影响转化的一系列因素如:pH、发酵时间、通气量、底物投料量及溶解性、细胞通透性进行控制,考察以上因素对羟化反应的影响,确定最佳转化条件.结果 该突变株培养基最佳配方为葡萄糖30 g/L、玉米浆20 g/L、蚕蛹粉5 g/L、硫酸铵2.5 g/L、磷酸氢二钾1 g/L、硫酸镁0.5 g/L和硫酸亚铁0.02 g/L.在pH 6.0~6.5、菌龄48~60 h、250 mL摇瓶装液量为30和40 mL、转化时间72 h时能取得最佳转化效果.在底物投料量为2 mg/mL时,采用4%无水乙醇溶解底物或加入0.75 mg/mL Tween 80均有利于羟化反应,在以上优化条件下转化率分别达到62.5%和66.8%.结论 突变株Metarhizium anisopliae M28-203能有效地在AD上引入11α羟基,为工业生产依普利酮及其他甾体药物提供关键中间体.%Objective To improve the transformation efficiency of androst-4-ene-3, 17-dione (AD) llα-hydroxylation mutant strain Metarhizium anisopliae M28-203 through regulations of nutrition metabolism and biotransformation conditions. Methods The optimal fermentation culture was determined by orthogonal test. The effects of pH, fermentation time, air flow, substrate addition method and substrate concentration on hydroxylation were investigated and the optimal transformation condition was constructed. Results The optimal fermentation culture contained glucose 30 g/L, corn steep 20 g/L, silkworm chrysalis powder 5 g/L, (NH4)2SO4 2.5 g/L, K2HPO4 1 g/L, MgSO4 0.5 g/L and FeSO4 0.02 g/L. The optimal transformation condition was pH 6.0 - 6.5, 48 - 60 h mycelium age, 30 and 40 mL medium volume in 250 mL shake flask and 72 hour transformation time. Both 4% ethanol and 0.75 mg/mL Tween 80 were helpful for substrate solubility, and then

  14. Efeitos de Beauveria bassiana (Bals Vuill e Metarhizium anisopliae (Metsc Sorok sobre fêmeas ingurgitadas de Amblyomma cajennense (Fabricius, 1787 em condições de laboratório Effects of Beauveria bassiana (Bals Vuill and Metarhizium anisopliae (Metsc Sorok on engorged females of Amblyomma cajennense (Fabricius, 1787 in laboratory conditions

    Directory of Open Access Journals (Sweden)

    R.C.S. Reis

    2004-12-01

    Full Text Available The in vitro susceptibility of Amblyomma cajennense engorged females to some isolated of the fungus Beauveria bassiana and Metarhizium anisopliae was verified and lethal concentrations (LC 50 and LC 90 were calculated. The females were dived in conidia suspensions for five minutes, and kept in climatically controlled chambers BOD under 27° C and 80% relative humidity. Each bioassay had four treatments in concentrations of 10(5,10(6,10(7e10(8 conidia/ml. A control group was also used. The following characteristics were evaluated: weight and period of oviposition, indexes of reproductive and nutritional efficiency and percentage of microbiological control. A dose dependent negative effect was observed in ticks treated with the suspension. All isolates tested cause a negative effect on in vitro tests of engorged females of A. cajennense, suggesting its potential for microbiological control of tick's species.

  15. Efficacy of Metarhizium anisopliae in controlling the two-spotted spider mite Tetranychus urticae on common bean in screenhouse and field experiments.

    Science.gov (United States)

    Bugeme, David Mugisho; Knapp, Markus; Ekesi, Sunday; Chabi-Olaye, Adenirin; Boga, Hamadi Iddi; Maniania, Nguya Kalemba

    2015-02-01

    The efficacy of aqueous and emulsifiable formulations of the fungus Metarhizium anisopliae isolate ICIPE78 was evaluated on the population density of Tetranychus urticae infesting common bean plants under screenhouse and field conditions. Synthetic acaricide abamectin was included as a check. Bean plants were artificially infested with T. urticae and allowed to multiply. Three treatments were applied in the screenhouse and 1 treatment in field trials. Mite density was recorded 2 d before spraying and weekly postspraying. The number of pods per plant, number of seeds per pod, and the dry weight of seeds per plant were recorded only in the screenhouse trials. In both screenhouse and field trials, fungal formulations applied at the concentration of 10(8) conidia/mL and the acaricide reduced the population density of mites as compared to the controls. There were significant differences in T. urticae population densities between the treatments at the various post-spraying sampling dates. In the screenhouse, the mite densities were near zero from 3-week postspraying in the treated leaves. At 4-week postspraying, there were no more leaves in the untreated control (T1) and in the control water + Silwet-L77 (T2). Fungal formulations were as effective as abamectin in reducing mite densities in both screenhouse and field experiments. There were significant differences in the production parameters during the 2 screenhouse trials, with fungal and abamectin treatments generally having the highest yield. Results of this study underline the potential of the M. anisopliae isolate ICIPE78 as an alternative to acaricides for T. urticae management.

  16. Patogenicidade, DL50 e TL50 de isolados de Metarhizium anisopliae (Metsch. Sorok. para o percevejo castanho das raízes Scaptocoris carvalhoi BECKER (Hemiptera: Cydnidae

    Directory of Open Access Journals (Sweden)

    Xavier Luciane Modenez Saldivar

    2005-01-01

    Full Text Available Objetivou-se neste trabalho avaliar a patogenicidade de isolados do fungo Metarhizium anisopliae em Scaptocoris carvalhoi BECKER, 1967, bem como determinar a Dose Letal média (DL50 e o Tempo Letal médio (TL50, em laboratório. Os experimentos foram conduzidos na Embrapa Agropecuária Oeste em Dourados, MS, durante 2003. Suspensões de quatro isolados de M. anisopliae (Ma7, Ma69, Ma283 e Ma342 foram preparadas nas concentrações 10(4, 10(5, 10(6, 10(7 e 10(8 conídios mL-1 e inoculados topicamente 5mil dessas suspensoes sobre S. carvalhoi correspondendo, respectivamente, a 50, 500, 5.000, 50.000 e 500.000 conídios percevejo-1. Após a inoculação, os insetos (10 adultos e cinco ninfas foram acondicionados em gerbox (parcela e mantidos em câmaras climatizadas reguladas para 26?1masculineC, UR 85%, sem fotofase. Na maior dose testada (500.000 conídios inseto-1, observou-se um incremento significativo de mortalidade do percevejo com o aumento da dose do fungo, para todos os isolados testados. Os menores valores da DL50 foram observados com os isolados Ma69 e Ma7 e o maior com Ma283. Os valores do TL50 variaram de 0,32 a 5,84 dias, sem diferirem significativamente, entre si. Os isolados Ma69 e Ma7 apresentam potencial para serem empregados no controle de S. carvalhoi a campo.

  17. 杨扇舟蛾幼虫高毒力绿僵菌菌株筛选研究%Studies on the screening of high virulent Metarhizium anisopliae against Clostera anachoreta larva

    Institute of Scientific and Technical Information of China (English)

    潘永胜; 徐福元; 韩正敏

    2012-01-01

    The growth rate and sporulation quantity of 21 Metarhizium anisopliae strains were measured in this paper,indicating that Metarhizium anisopliae1(Ma1,Ma42,Ma788,Ma789)had high yield of spore.And the germination percentage with different temperatures in water bath treatment and toxicity of different M.anisopliae strains against Clostera anachoreta were measured.The results showed that Ma789 was the most virulent one.%测定了21株绿僵菌的平板单菌落生长和产孢量情况,证明(Ma1,Ma42,Ma788,Ma789)4菌株产孢量高,通过对这4株绿僵菌菌株的不同温度水浴处理,发现了孢子的萌发条件;并开展了4菌株对杨扇舟蛾幼虫的毒力测定,筛选出Ma789菌株的致病力较强。

  18. 金龟子绿僵菌(Metarhizium anisopliae HN1)几丁质酶基因的克隆及高效表达%Cloning Chitinase Gene of the Entomopathogene Fungus Metarhizium anisopliae HN1 and High-level Expression in Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    任文彬; 张世清; 黄俊生

    2006-01-01

    几丁质酶是昆虫病原真菌金龟子绿僵菌致病力的主要因子之一.用RT-PCR方法,从分离筛选到的高毒力金龟子绿僵菌Metarhizium anisopliae HN1中,扩增得到几丁质酶基因全长,此基因全长为1275bp,登录号为DQ011865,经Blastn分析此基因序列与M. anisopliae E6的chi1基因(AF02749)同源率为96%.以pET-22b(+)为基础载体,构建pET-chi重组表达载体,在大肠杆菌(Escherichia coli)BL 21中进行表达.经SDS-PAGE分析,获得了42kDa大小的重组目的蛋白,目的蛋白占表达总蛋白含量的63.3%.菌体经冷冻与超声波破碎后,按DNS法可测得几丁质酶的活性.

  19. 金龟子绿僵菌(Metarhizium anisopliae HN1)几丁质酶与谷胱甘肽S-转移酶GST在大肠杆菌中的高效融合表达%High-efficient Fusion Expression of Chitinase and GST from Metarhizium anisopliae HN1 in Escheichia coli

    Institute of Scientific and Technical Information of China (English)

    任文彬; 张世清; 黄俊生

    2009-01-01

    使用RT-PCR方法,从高毒力金龟子绿僵菌Metarhizium anisopliae HN1中,克隆得到一个全长为1 275 bp的几丁质酶基因,经Blast分析此基因序列与M. anisopliae E6的chi1基因(AF02749)同源率为96%.将此基因克隆到pGEX-6p-1载体上,使之与载体上一个约26 kD大小的谷胱甘肽S-转移酶(GST)相连,构建pGEX-chi融合表达载体,转化到大肠杆菌(Escherichia coli)BL 21中,经SDS-PAGE结果分析显示:表达出的融合蛋白大小为68 kD,此目的蛋白占表达总量的64.5%.经破碎处理后可检测到几丁质酶活性.

  20. 金龟子绿僵菌致病的分子机理研究进展%Research Progress on Pathogenic Molecular Mechanism of Metarhizium anisopliae

    Institute of Scientific and Technical Information of China (English)

    刘颖; 殷从松

    2010-01-01

    金龟子绿僵菌(Metarhizium anisopliae)是一类重要的昆虫病原真菌,在害虫生物防治中起着重要作用.与化学农药相比,昆虫病原真菌开发的真菌杀虫剂作用时间持久、无环境污染、对非靶标生物安全,但存在致死时间长,杀虫效率低,防效不稳定等缺点,严重阻碍了真菌杀虫剂的广泛应用,为解决这一问题,从分子水平揭示昆虫病原真菌的致病机理尤为重要.从分子水平阐述了金龟子绿僵菌对寄主的识别与粘附、绿僵菌侵染寄主过程中附着胞的形成、绿僵菌对寄主体壁的穿透机制和绿僵菌进入寄主血淋巴后的适应机制.

  1. Production of chitosanase by Metarhizium anisopliae in solid state fermentation%金龟子绿僵菌固态发酵产壳聚糖酶

    Institute of Scientific and Technical Information of China (English)

    刘桦; 张涛; 王玉明

    2007-01-01

    对金龟子绿僵菌(Metarhizium anisopliae)AS3.4606产壳聚糖酶进行了固态发酵条件优化及酶学部分特征的研究.正交实验结果表明,以麸皮为碳源,日本根霉菌丝体粉末为氮源,在起始pH 5.0,m(碳):m(氨)为1∶4,m(干重):m(液体)为1:1.4,27 ℃下培养120 h,壳聚糖酶活性可达35.08 U/g(干培养基).粗酶液的最适反应温度为40 ℃,最适反应pH为5.0,酶在pH 5.0~6.0条件下稳定性最高.该酶不能水解固体甲壳素和纤维素粉末,最适底物为胶体壳聚糖.

  2. Destruxin B Isolated from Entomopathogenic Fungus Metarhizium anisopliae Induces Apoptosis via a Bcl-2 Family-Dependent Mitochondrial Pathway in Human Nonsmall Cell Lung Cancer Cells

    Directory of Open Access Journals (Sweden)

    Chun-Chi Wu

    2013-01-01

    Full Text Available Destruxin B, isolated from entomopathogenic fungus Metarhizium anisopliae, is one of the cyclodepsipeptides with insecticidal and anticancer activities. In this study, destruxin B was extracted and purified by ion-exchange chromatography, silica gel chromatography, and semipreparative high-performance liquid chromatography. The potential anticancer effects and molecular mechanisms of destruxin B in human nonsmall cell lung cancer cell lines were characterized. Our results showed that destruxin B induced apoptotic cell death in A549 cells. This event was accompanied by the activation of caspase-2, -3, and -9. Moreover, destruxin B increased the expression level of proapoptotic molecule, PUMA, while decreased antiapoptotic molecule Mcl-1. Additionally, the translocation of Bax from cytosol to mitochondrial membrane was observed upon destruxin B treatment. Knockdown of Bax by shRNA effectively attenuated destruxin-B-triggered apoptosis in A549 cells. Interestingly, similar toxic effects and underlying mechanisms including caspase activation, upregulation of PUMA, and downregulation of Mcl-1 were also observed in a p53-null lung cancer H1299 cell line upon destruxin B treatment. Taken together, our findings suggest that destruxin-B-induced apoptosis in human nonsmall cell lung cancer cells is via a Bcl-2 family-dependent mitochondrial pathway.

  3. 11-αhydroxylation Process of Steroids with Metarhizium anisopliae%金龟子绿僵菌对甾体底物11-α羟化反应的工艺

    Institute of Scientific and Technical Information of China (English)

    刘玲玲; 杜大庆; 张星元; 赵允麟

    2006-01-01

    研究了金龟子绿僵菌(Metarhizium anisopliae)对甾体底物16α、17α-环氧黄体酮(16α,17α-epoxy-4-pregnene-3,20-dione)的羟化反应工艺,考察了培养基组成、接种量、投料时间、转化时间、溶料方式、发酵级数等因素,探索了稀释发酵液以提高转化效率的新工艺.

  4. Effects of single and combined applications of entomopathogenic fungi and nematodes against Rhynchophorus ferrugineus (Olivier)

    Science.gov (United States)

    This study was carried out to investigate the insecticidal properties of Beauveria bassiana and Metarhizium anisopliae, and Heterorhabditis bacteriophora Poinar for their virulence against 2nd, 4th and 6th instar larvae of Rhynchophorus ferrugineus (Olivier). Both fungi were either applied alone or ...

  5. SUBSTRATO DE CRESCIMENTO DO FUNGO Metarhizium anisopliae INFLUENCIA NA VIRULÊNCIA CONTRA LARVAS DE Aedes aegypti

    Directory of Open Access Journals (Sweden)

    Aline Teixeira Carolino

    2016-11-01

    Full Text Available O presente estudo investigou a virulência dos isolados 4556 e ESALQ 818 do fungo M. anisopliae crescidos nos substratos arroz parboilizado e nos meios de cultivo BDA (para o isolado 4556 e SDA (para o isolado ESALQ 818. Ensaios de virulência foram realizados com larvas de Aedes aegypti provenientes de ovos coletados no campo. Os ovos foram colocados na água e deram origem as larvas que foram utilizadas nos experimentos. Os conídios de M. anisopliae (ambos isolados crescidos nos diferentes substratos foram formulados em Tween 80 e utilizados na concentração de 1x107 conídios ml-1. O estudo constatou que os dois isolados crescidos no arroz parboilizado foram mais virulentos para larvas de campo de A. aegypti. Apenas 7,5% das larvas tratadas com os conídios do isolado 4556 crescidos no arroz sobreviveram e no meio de cultivo BDA a sobrevivência das larvas foi de 65,%, após 24 horas de infecção. As larvas tratadas com conídios do isolado ESALQ 818 produzidos no arroz apresentaram 52,5% de sobrevivência e as tratadas com conídios produzidos no meio SDA apresentaram sobrevivência de 85%, após 24 horas de infecção.

  6. Occurence of thrips on Niagara table grape and its control with the insecticides thiacloprid and methiocarb associated with Metarhizium anisopliae Ocorrência de tripes em uva Niagara e seu controle com os inseticidas thiacloprid e methiocarb associados com Metarhizium anisopliae

    Directory of Open Access Journals (Sweden)

    ROGÉRIO BIAGGIONI LOPES

    2002-04-01

    Full Text Available Thrips are reported as important pests on table grapes in United States and several countries of Europe. Damage caused by thrips, particulary Frankliniella occidentalis, was observed on niagara table grape crop in Limeira-SP, Brazil. During the blooming period, high thrips densities were observed feeding on pollen and small berries. The symptoms left were more visible after the development of the berries and were characterized by dark scars and suberized surface on berries, sometimes causing the berry to crack, and the seed to prolapse. The effect of insecticides thiacloprid or methiocarb, associated or not with the entomopathogenic fungus Metarhizium anisopliae were evaluated during the blooming period. For evaluation of thrips damage on fruits, the treatments were applied three additional times, 7, 14 and 21 days after the first application. The treatments were: a M. anisopliae (strain 1037 1x10(7 conidia/mL; b thiacloprid 20mL/100L; c-d methiocarb 100 and 150mL/100L; e methiocarb 100mL/100L + M. anisopliae 1x10(7 conidia/mL. Only methiocarb, associated or not with the fungus, was effective in reducing thrips infestation, and no phytotoxic damage was observed. The efficiency of methiocarb 150mL/100L and the insecticide associated with the fungus for the control of the thrips population was 84.2 and 95.5%, respectively. In both cases, there was a reduction of approximately 70% in the number of berries with scars symptoms. For control of thrips on table grapes, chemical insecticides associated or not with M. anisopliae should be applied during the blooming period of the crop.Os tripes são mencionados como importantes pragas da cultura da uva de mesa em diversos países da Europa e Estados Unidos. Em área de cultivo da uva Niagara localizada na região de Limeira-SP verificou-se a ocorrência de danos provocados por Frankliniella occidentalis. Essa praga foi observada, principalmente, durante a fase de florescimento, alimentando-se de pólen e

  7. Purification and properties of chitinase from Metarhizium anisopliae%绿僵菌几丁质酶的分离纯化及性质

    Institute of Scientific and Technical Information of China (English)

    杨革; 陈洪章; 李佐虎

    2005-01-01

    从自然罹病死亡的金龟子体内分离到一株金龟子绿僵菌(Metarhizium anisopliae),它在几丁质的诱导下能产生较高活性的几丁质酶.发酵液经硫酸铵盐析、DEAE纤维素柱层析、Phenyl SepharoseTM 6 Fast Flow疏水柱层析等方法,得到电泳纯的几丁质酶.用SDS-PAGE测得该酶相对分子质量为61.5 kD,而经质谱分析为57.14 kD.最适反应温度为55 ℃,最适反应pH值为6.0,酶的等电点pI为4.02,其N末端序列为VIGPAAPL,用硫酸-酚法测得其含糖量为56.2%.水解几丁质的Km为14.5 μmol·L-1.该酶在45 ℃,pH值3.0~9.5较为稳定.Zn2+、Ca2+、Ba2+和Mn2+离子对几丁质酶活性有明显的促进作用,而Hg2+、Co2+和Fe2+离子完全抑制几丁质酶的活性.此酶还可被EDTA所抑制,表明金属离子为其活性所必需.PMSF试剂对几丁质酶的活力影响比较大,丝氨酸可能是酶活力的必需基团.

  8. Screening of Metarhizium anisopliae Strain with High Virulence against Larvae of Curculio chinensis(Coleoptera:Curculionidae)%感染油茶象幼虫的高致病力金龟子绿僵菌菌株筛选

    Institute of Scientific and Technical Information of China (English)

    何学友; 蔡守平; 杜月飞; 陈德兰; 黄金水; 李孔泉

    2015-01-01

    Objective]Curculio chinensis ( Coleoptera: Curculionidae ) is one of the most serious pests of oil-tea Camellia fruits. Screening of Metarhizium anisopliae strains with high virulence against this pest is important for using fungi as biocontrol agent to suppress this pest.[Method]Colony growth and sporulation of 7 strains of M. anisopliae were measured and their virulence against C. chinensis larvae were investigated in laboratory by using two inoculating methods ( Being dipped on larvae with conidia suspension of 107 spores·mL -1 and being mixed in soil with 105 spores·g -1 ) . The infection effects in field were evaluated through spraying conidia suspension of two superior strains. [Result]Bioassay results showed that larvae of C. chinensis were susceptible to different M. anisopliae strains. The larval mortalities reached 100% on the 13 th day after inoculation using dipping inoculative method and the 9 th day for conidia-soil mixing inoculative method. The median lethal time ( LT50 ) of dipping inoculative method was 1. 65 days to 4. 26 days,and that of conidia-soil mixing inoculative method was 1. 96 days to 3. 51 days. The FJMa201101 and FJMa201205 strains of M. anisopliae were the most virulent against larvae of C. chinensis. When larvae inoculated with FJMa201101,the LT50 was 1. 65 days,and the infection rate was 86. 6% when using dipping inoculative method,while the LT50 was 2. 11 days and the infection rate was 94. 5% respectively for conidia-soil mixing inoculative method. When inoculated with FJMa201205,the LT50 was 1. 71 days with the infection rate of 91. 1% by using dipping inoculative method,whilst it was 1. 96 days with the infection rate of 88. 9% with conidia-soil mixing inoculative method. In field test,there was no significant difference in infection rate between the two superior strains,but the infection rates of larvae introduced to soil after spraying conidia suspension were significantly higher than those introduced before spraying

  9. Community composition and population genetics of insect pathogenic fungi in the genus Metarhizium from soils of a long-term agricultural research system.

    Science.gov (United States)

    Kepler, Ryan M; Ugine, Todd A; Maul, Jude E; Cavigelli, Michel A; Rehner, Stephen A

    2015-08-01

    Fungi in the genus Metarhizium are insect pathogens able to function in other niches, including soil and plant rhizosphere habitats. In agroecosystems, cropping and tillage practices influence soil fungal communities with unknown effects on the distribution of Metarhizium, whose presence can reduce populations of crop pests. We report results from a selective media survey of Metarhizium in soils sampled from a long-term experimental farming project in the mid-Atlantic region. Field plots under soybean cultivation produced higher numbers of Metarhizium colony-forming units (cfu) than corn or alfalfa. Plots managed organically and via chisel-till harboured higher numbers of Metarhizium cfu than no-till plots. Sequence typing of Metarhizium isolates revealed four species, with M. robertsii and M. brunneum predominating. The M. brunneum population was essentially fixed for a single clone as determined by multilocus microsatellite genotyping. In contrast, M. robertsii was found to contain significant diversity, with the majority of isolates distributed between two principal clades. Evidence for recombination was observed only in the most abundant clade. These findings illuminate multiple levels of Metarhizium diversity that can be used to inform strategies by which soil Metarhizium populations may be manipulated to exert downward pressure on pest insects and promote plant health. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  10. Spatial separation of semiochemical Lurem-TR and entomopathogenic fungi to enhance their compatibility and infectivity in an autoinoculation system for thrips management

    NARCIS (Netherlands)

    Mfuti, D.K.; Subramanian, S.; Tol, van R.W.H.M.; Wiegers, G.L.; Kogel, de W.J.; Niassy, S.; Plessis, du H.; Ekesi, S.; Maniania, N.K.

    2016-01-01

    BACKGROUND
    The effect of spatial separation of the semiochemical Lurem-TR, which has been found to inhibit conidia of entomopathogenic fungi when put together, on the persistence of conidia of Metarhizium brunneum and M. anisopliae was evaluated in the greenhouse and field in order to develop

  11. Infection of the malaria mosquito, Anopheles gambiae, with two species of entomopathogenic fungi: effects of concentration, co-formulation, exposure time and persistence

    NARCIS (Netherlands)

    Mnyone, L.L.; Kirby, M.J.; Lwetoijera, D.W.; Mpingwa, M.W.; Knols, B.G.J.; Takken, W.; Russell, T.L.

    2009-01-01

    Background - Entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana isolates have been shown to infect and reduce the survival of mosquito vectors. Methods - Here four different bioassays were conducted to study the effect of conidia concentration, co-formulation, exposure time and per

  12. Entomopathogenic fungi Metarhizium spp. in the soil environment of an agroecosystem

    DEFF Research Database (Denmark)

    Steinwender, Bernhardt Michael

    Species of the entomopathogenic fungal genus Metarhizium are found worldwide predominantly in the soil environment where they infect a broad spectrum of insects, but also associate with plant roots. To increase performance of Metarhizium as biological control agents against pests, fundamental...

  13. Cloning and Sequence Analysis of Mannose-6-phosphate lsomerase cDNA from Metarhizium anisopliae ZJ1109%金龟子绿僵菌甘露糖6-磷酸异构酶基因cDNA序列的克隆及分析

    Institute of Scientific and Technical Information of China (English)

    李亚

    2011-01-01

    The primers were designed according to the conservative sequence of mannose-6-phosphate isomerase gene, and the complete cDNA sequence of Metarhizium anisopliae was amplified by RT-PCR and RACE PCR. The complete cDNA sequence of mpi gene was 1513 bp and the complete ORF length was 1328 bp which encoded a protein with 441 amino acid residues. Blast analysis indicated the deduced amino acid sequence of the mpi gene from M. Anisopliae showed high homology with other fungi respectively. The analysis of protein structure showed MPI protein had the characteristic of the conservative phosphate enzyme, and mainly constructed by a-helix and random coil.%通过绿僵菌属甘露糖6-磷酸异构酶基因保守核苷酸区域设计简并性引物,采用RT-PCR及RACE-PCR技术成功克隆了金龟子绿僵菌mpi基因cDNA序列.该基因cDNA序列全长为1513bp,开放阅读框长度为1328 bp,共编码441氨基酸.BLAST分析发现该基因演绎的氨基酸序列与其它真菌同源性较高,蛋白结构分析表明MPI蛋白是较保守的蛋白磷酸酶结构特征,主要由α螺旋和不规则卷曲构成.

  14. Impacts of 6 fungicides on conidial germination of Metarhizium anisopliae%6种杀菌剂对金龟子绿僵菌孢子萌发的影响

    Institute of Scientific and Technical Information of China (English)

    叶小真; 陈全助; 宋漳; 陈耀群

    2012-01-01

    The inhibition of six fungicides on the germination of conidia of Metarhizium anisopliae were determined. The results showed that different fungicides had different inhibition on conidia of M. Anisopliae, and the inhibition decreased with declining concentration of fungicide. Among them, myclobutanil showed minimum inhibition on M. Anisopliae. Except the strain Mal03, the inhibition on other strains were less than 35%. The average inhibition rates of zineb on the stains Ma202, Ma201 and Ma203 were low. Other fungicides had obvisous inhibition on most strains of M. Anisopliae, especially chlorothalonil and thiophanate-methyl. The average inhibition rates of these two fungicides on the strains Mal03, Ma205, Ma207 and Ma208 were 100%. They also effectively inhibited the other strains. Therefore, we suggested that chlorothalonil, thiophanate-methyl and M. Anisopliae should not be applied in the field at the same time and place. Otherwise, the control effect of M. Anisopliae will be significantly influenced by these two fungicides.%测定了6种杀菌剂对金龟子绿僵菌分生孢子萌发的抑制作用.结果表明,不同杀菌剂对金龟子绿僵菌的抑制作用不同,且随着杀菌剂浓度的下降而减小.其中以腈菌唑的抑制效果最弱,除Ma103菌株外,对其他菌株抑制率均低于35%.代森锌对Ma202,Ma201和Ma203菌株的平均抑制率都较小.其他几种杀菌剂都明显抑制绿僵菌大部分菌株分生孢子的萌发,特别是百菌清和甲基托布津对Ma103,Ma205,Ma207和Ma208菌株的平均抑制率都为100%.2种药剂也强烈抑制其他菌株分生孢子的萌发.因此,应该尽量避免百菌清和甲基托布津与金龟子绿僵菌同时同地使用,否则会明显影响金龟子绿僵菌制剂的应用效果.

  15. Compatibility of Eighteen Chemical Pesticides with Metarhizium anisopliae%18种化学农药与绿僵菌相容性研究

    Institute of Scientific and Technical Information of China (English)

    张英财; 农向群; 张泽华; 王广君

    2012-01-01

    本文检测了18种常用低毒化学农药对生防真菌绿僵菌Metarhizium anisopliae孢子萌发、菌丝生长及产孢的影响,分析它们与绿僵菌的相容性。总体看,杀虫剂、植物生长调节剂和除草剂与绿僵菌的相容性好于杀菌剂。其中,氰戊菊酯与绿僵菌相容性最好,即使在10倍的推荐浓度下,对孢子萌发无影响,对菌丝生长速度和产孢量的抑制率只有25.13%和38.63%。高效氯氟氰菊酯和乐果与绿僵菌也有较好的相容性,在推荐的田间使用浓度下,孢子萌发抑制率为30%左右,对菌丝生长和产孢量基本没有抑制。溴氰菊酯、啶虫脒、吡虫啉、甲霜灵、氟乐灵、阿维菌素、敌百虫、毒死蜱和波尔多液与绿僵菌有一定的相容性。矮壮素、哒螨灵、多菌灵、甲基硫菌灵、代森锰锌和三唑酮与绿僵菌相容性很差。%Impacts of eighteen low-toxicity commercial chemical pesticides on conidial germination,mycelial growth and sporulation of Metarhizium anisopliae,an entomopathogenic fungus as biocontrol agent,were tested to evaluate compatibility between the fungus and the pesticides.Results showed that the tested insecticides,plant growth regulators and herbicides were more compatible with the fungus than the tested germicides.Among them,deltamethrin showed the highest compatibility with the fungus.Even at ten times of recommended concentration,it had no inhibition on conidial germination and only inhibited mycelial growth by 25.13% and sporulation by 38.63%.Lambda-cyhalothrin and dimethoate demonstrated moderate compatibility with the fungus.Their inhibition rates on conidial germination were about 30%,but no inhibition on mycelial growth at the recommended concentration.Deltamethrin,acetamiprid,imidacloprid,metalaxyl,trifluralin,abamectin,trichlorphon,chlorpyrifos and bordeaux mixture showed lower compatibility with the fungus,while chlormequa,pyridaben,carbendazim,thiophanate-methyl,mancozeb and

  16. Responsiveness of entomopathogenic fungi to menadione-induced oxidative stress.

    Science.gov (United States)

    Azevedo, Rosana F F; Souza, Roberta K F; Braga, Gilberto U L; Rangel, Drauzio E N

    2014-12-01

    Entomopathogenic fungi are predisposed to ROS induced by heat and UV-A radiation when outside the insect host. When inside the host, they are subject to phagocytic cells that generate ROS to eliminate invading pathogens. The oxidative stress tolerance of the entomopathogenic fungi Aschersonia aleyrodis (ARSEF 430 and 10276), Aschersonia placenta (ARSEF 7637), Beauveria bassiana (ARSEF 252), Isaria fumosorosea (ARSEF 3889), Lecanicillium aphanocladii (ARSEF 6433), Metarhizium acridum (ARSEF 324), Metarhizium anisopliae (ARSEF 5749), Metarhizium brunneum (ARSEF 1187 and ARSEF 5626), Metarhizium robertsii (ARSEF 2575), Tolypocladium cylindrosporum (ARSEF 3392), Tolypocladium inflatum (ARSEF 4877), and Simplicillium lanosoniveum (ARSEF 6430 and ARSEF 6651) was studied based on conidial germination on a medium supplemented with menadione. Conidial germination was evaluated 24 h after inoculation on potato dextrose agar (PDA) (control) or PDA supplemented with menadione. The two Aschersonia species (ARSEF 430, 7637, and 10276) were the most susceptible fungi, followed by the two Tolypocladium species (ARSEF 3392 and 4877) and the M. acridum (ARSEF 324). Metarhizium brunneum (ARSEF 5626) and M. anisopliae (ARSEF 5749) were the most tolerant isolates with MIC 0.28 mM. All fungal isolates, except ARSEF 5626 and ARSEF 5749, were not able to germinate at 0.20 mM.

  17. Microbial control of the invasive spiraling whitefly on cassava with entomopathogenic fungi

    OpenAIRE

    Boopathi, Thangavel; Karuppuchamy,Palaniappan; Singh,Soibam B.; Kalyanasundaram, Manickavasagam; S. Mohankumar; Ravi,Madhaiyan

    2015-01-01

    Abstract The entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, Lecanicillium lecanii and Isaria fumosorosea were tested for their efficacy in managing the exotic spiraling whitefly Aleurodicus dispersus (Hemiptera, Aleyrodidae) on cassava (Manihot esculenta) during 2 seasons (2011-2012 and 2012-2013). The fungi I. fumosorosea and L. lecanii exhibited promising levels of control (> 70% mortality of the A. dispersus population). The percent mortality increased over time in both...

  18. Abundance of soil-borne entomopathogenic fungi in organic and conventional fields in the Midwestern USA with an emphasis on the effect of herbicides and fungicides on fungal persistence

    Science.gov (United States)

    Naturally-occurring entomopathogenic fungi provide a valued service of killing agricultural pests and subduing pest outbreaks. Species such as Beauveria bassiana and Metarhizium anisopliae have been researched in many experiments and have proven to be effective pathogens. This research will focus ...

  19. Impacts of chemical insecticides on extracellular protease and chitinase activities of Metarhizium anisopliae%化学杀虫剂对绿僵菌胞外蛋白酶和几丁质酶活性的影响

    Institute of Scientific and Technical Information of China (English)

    宋漳; 叶小真

    2010-01-01

    研究了几种化学杀虫剂对金龟子绿僵菌(Metarhizium anisopliae)胞外蛋白酶和几丁质酶活性的影响.结果表明,化学杀虫剂对绿僵菌的胞外蛋白酶和几丁质酶活性影响较大,供试的12株金龟子绿僵菌大部分菌株的胞外蛋白酶和几丁质酶活性均明显降低,仅有Ma202、Ma207菌株的胞外蛋白酶和几丁质酶活性略有提高.

  20. 金龟子绿僵菌发酵代谢产物中苦马豆素的初步研究%Preliminary Study of Swainsoine from Metabolism Products of the Metarhizium anisopliae

    Institute of Scientific and Technical Information of China (English)

    路宏朝; 杨鸣琦; 朱晓飞; 何欣; 李卫彬

    2006-01-01

    金龟子绿僵菌(Metarhizium anisopliae)经过发酵培养,采用气相色谱法对发酵代谢产物中的苦马豆素(Swainsonine SW) 进行了定性和定量分析;同时用正丁醇萃取法对发酵液进行了初步的分离提取及薄层层析法检测.结果表明,金龟子绿僵菌发酵代谢产物中含有苦马豆素,且含量为10.2 131 mg/L;正丁醇萃取法是从发酵液中提取苦马豆素的有效方法.

  1. Avaliação da patogenicidade de Beauveria bassiana e Metarhizium anisopliae, sobre os diferentes estádios de Rhipicephalus sanguineus no cão e no ambiente

    OpenAIRE

    2007-01-01

    O presente trabalho teve como objetivo avaliar “in vitro”, a ação de Beauveria bassiana e Metarhizium anisopliae, para larvas, ninfas e adultos machos e fêmeas não alimentadas de Rhipicephalus sanguineus e para onde as mesmas fases do ciclo de vida do ácaro mantidas alimentadas em cães por meio de câmaras de alimentação coladas no dorso do animal. A ação no ambiente habitado pelo cão foi avaliada por meio de infestações naturais em boxes com todos os estádios do ácaro. Foram usadas suspensões...

  2. Entomopathogenic fungi Metarhizium spp. in the soil environment of an agroecosystem

    DEFF Research Database (Denmark)

    Steinwender, Bernhardt Michael

    ecological knowledge of Metarhizium is necessary. The present PhD project contributed to this knowledge, particularly of Metarhizium spp. occurrence and abundance within a single Danish agroecosystem, with emphasis on the molecular diversity and ecological traits. Metarhizium was isolated from bulk soil...... several sympatric species and genotypes. The isolated species and their genotypes were evaluated for ecological traits including UVB tolerance, temperature dependent in vitro growth, virulence and conidia production on infected cadavers, and mycelial growth from insect cadavers into the surrounding soil....... The results showed a broad variety of ecological characteristics of the different genotypes, indicating their potential adaptations to different ecological niches within the agroecosystem. The presented knowledge of co-existence of diverse Metarhizium species and genotypes will be valuable for the development...

  3. Potential of Metarrhizium anisopliae and Beauveria bassiana isolates and Neem oil to control the aphid Lipaphis erysimi (Kalt.) (Hemiptera: Aphididae); Potencial de isolados de Metarhizium anisopliae e Beauveria bassiana e do oleo de Nim no controle do pulgao Lipaphis erysimi (Kalt.) (Hemiptera: Aphididae)

    Energy Technology Data Exchange (ETDEWEB)

    Araujo Junior, Jose M. de; Marques, Edmilson J.; Oliveira, Jose V. de [Universidade Federal Rural de Pernambuco (UFRPE), Recife, PE (Brazil). Dept. de Agronomia - Entomologia], e-mail: jma_junior@yahoo.com.br, e-mail: emar@depa.ufrpe.br, e-mail: vargasoliveira@uol.com.br

    2009-07-15

    This work aimed to determine the efficiency of the entomopathogenic fungi Metarrhizium anisopliae and Beauveria bassiana to control the aphid Lipaphis erysimi (Kalt.) (Hemiptera:Aphididae) in kale Brassica oleracea var acephala D.C., as well as their compatibility with a neem oil formulation (Neemseto{sup R}). Ten isolates of both fungi were tested and the most pathogenic ones were B. bassiana CG001 and M. anisopliae CG30 with 90% and 4.4 days, and 64% and 3.8 days of mortality and median lethal time, respectively. Bioassays with neem at concentrations of 0.5, 1.0 and 2.0% were done either by leaf discs dipping or spraying the aphids on the leaf discs. The neem spraying treatment at 2.0% provided 90% mortality. The use of B. bassiana isolate CG001 or M. anisopliae isolate CG30 with neem at 0.125, 0.25, and 0.5%, demonstrated that these isolates could have their spore viability or colony growth affected when exposed to neem concentrations higher than 0.25%. In absolute values, the isolates B. bassiana CG001 and M. anisopliae CG30 are the most virulent to L. erysimi, and could be utilized in the management of this pest. (author)

  4. Influence of pesticides used potatoes control on the growth of entomopathogenic fungi isolated from potatoes fields

    Directory of Open Access Journals (Sweden)

    Ryszard Miętkiewicz

    2013-12-01

    Full Text Available The growth of Metarhizium anisopliae, M.flavoviridae and Paecilomyces fumosoroseus was estimated on Sabouraud's medium to which insecticides and herbicides were added in three doses: A - 10 times higher from recommended field dose, B - as recommended field dose, C - 10 timer lower than recommended. Fungicides were used in B and C doses as well as in dose D - 100 times lower than recomended one. The fungi were obtained from soil under potatoes using Galleria mellonella as bait insect. Chlorothalonil and copper oxychloride were chosen from fungicides, linuron, MCPA, fluazifop-P-butyl and dikwat - from herbicides and deltamethrin, teflubenzuron and fozalon from insecticides. The growth of both species of Metarhizium was stronger inhibited than of Paecilomyces fumosoroseus by fungicides however colonies of Metarhizium always overpassed 50% controls colonies apart from M. flavoviridae on medium with copper oxychloride at concentration B. Herbicide linuron was more toxic to fungi than fungicides. M. anisopliae and M. fluvoviridae did not grow on medium containing dose A and dose B this herbicide but the colonies of P. fumosoroseus at dose B did not overpass 20% of controlled ones. MCPA and fluazifop-P-butyl inhibited fungal colonies in approximated way. On the medium with these herbicides in concentration A fungal colonies were strongly inhibited and the growth of fungi on medium with MCPA at this concentration appeared not before 5 days after inoculation. Dikwat in dose A strongly inhibited the growth of M. anisopliae but in remaining combinations growth of fungal colonies was similar to controlled ones. Fozalon, among insecticides, inhibited the growth of inwestigated fungi strongest. On the medium containing this insecticide in dose A all fungi did not grow, and in dose B colonies of both species of Metarhizium did not overpass 40% of controlled ones. Deltamethrin in dose A and B inhibited the growth of M. anisopliae and M. flavoviridae, but

  5. Kinetic study of liquid fermentation for Metarhizium anisopliae IMI330189%金龟子绿僵菌IMI330189液体发酵动力学研究

    Institute of Scientific and Technical Information of China (English)

    王乔; 王海胜; 农向群; 张泽华

    2012-01-01

    The liquid fermentation kinetics of Metarhizium anisopliae IMI330189 was observed. Kinetic models of biomass growth and substrate consumption were established based on Sigmoid model and the parameters of the models were determined by Origin7.S software. The results showed that the models fitted well with the liquid fermentation process of M. anisopliae IMI330189. The maximum specific growth rate was found to be 0.084h' at 22.8h, specific total sugar consumption rate 0.246h-1 at 9.6h and specific total nitrogen consumption rate 0.007h-1 at 10.3h. The maximum yieldcoefficient of biomass from total sugar was 0.861g/g at 39.8h. The model simulation matched well with the experimental observations, which made it possible to elucidate the kinetic characteristics of M anisopliae IMI330189 during liquid fermentation. These models thus can be employed for the development and optimization of M. anisopliae IMI330189 liquid fermentation processes.%研究了金龟子绿僵菌IMI330189的液体发酵动力学.利用Sigmoid函数构建了该菌株液体发酵过程中的菌体生长和底物消耗的动力学模型,并运用Origin7.5软件拟合求解出各模型参数.结果表明,模型能够较好地拟合绿僵菌IMI330189液体发酵过程,其比生长速率在发酵第22.8h达到最大值,为0.084h-1;总糖比消耗速率在第9.6h达到最大值,为0.246h-1;总氮比消耗速率在第l0.3h达到最大值,为0.007h-1;菌体对总糖的得率系数在39.8h达到最高,为0.861g/g.模型拟合和实验数据具有良好的适应性,基本反映了绿僵菌IMI330189液体发酵过程的动力学特征,为其液体发酵工艺的优化和发展奠定了基础.

  6. 金龟子绿僵菌诱发UV-B抗性导致分生孢子产孢和孢子逆境耐受性的转化%Inducing UV-B tolerance of Metarhizium anisopliae var.anisopliae conidia results in a trade-off between conidial production and conidial stress tolerance

    Institute of Scientific and Technical Information of China (English)

    德罗奇欧· 兰格尔; Donald W Roberts

    2007-01-01

    The conidial tolerance of Metarhizium anisopliae var.anisopliae isolate ARSEF 2575 to UV-B irradiation is greatly influenced by growth-environment alterations. In this review, we report high variability in conidial UV-B tolerance in response to altered culture conditions.Conidia produced on insect cadavers[Zophobas morio(Coleoptera) or Galleria mellonella(Lepidoptera)] had low tolerance to UV-B radiation; and conidia produced on potato dextrose agar supplemented with yeast extract (PDAY) had medium UV-B tolerance; whereas conidia produced on a minimal medium without any carbon source (MM), on MM with a non-preferred carbon source such as lactose (=MML), on PDAY plus 1 M NaCl or KCl, or PDBY with high alkalinity had the highest UV-B tolerances. All of the above conditions that induced high UV-B tolerance, however, also greatly reduced conidial production. Comparisons between stress tolerance and conidial production, particularly with conidia produced under osmotic and nutritive stress, point out that the benefits of producing very tolerant conidia have the enormous cost of low conidial production. Growth under visible light also greatly improved conidial UV-B tolerance, but light did not negatively influence conidial production. Therefore, culture on rich media under light is proposed as the most promising approach to producing conidia with improved UV-B tolerance for biological control of pest insects in agriculture.

  7. Isolation and identification of Metarhizium anisopliae from natural infections of Coconut hispid beetle Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae) , and preliminary studies on against this pest in Hainan island, China%海南椰心叶甲病原菌金龟子绿僵菌的分离、鉴定及其生防潜力

    Institute of Scientific and Technical Information of China (English)

    詹儒林; 覃伟权; 宋妍; 张世清; H H HO; 许天委; 黄俊生

    2007-01-01

    椰心叶甲[Brontispa longissima(Gestro)]是椰子的重要害虫,近年来,该虫在海南岛发生普遍,椰子受害严重.由于椰心叶甲受到自然界中某些致病微生物的侵袭,在受害的椰子树心叶上常可发现椰心叶甲僵虫,并发现大部分僵虫表面长出了霉菌,本研究的目的在于从椰心叶甲僵虫表面的霉菌中分离出绿僵菌,并对分离菌株进行鉴定和致病性测定.从僵虫表面刮下孢子或菌丝体,置于绿僵菌选择性培养基(DOA)上培养,挑出真菌菌落,经纯化后,进行生物学特性、菌落生长速率及产孢量的测定,并从PPDA、OMA、V8A和PDA中筛选菌落生长及产孢最适培养基,同时对所分离的菌株进行对椰心叶甲的致病性测定.结果表明,所有分离菌株均鉴定为金龟子绿僵菌[Metarhizium anisopliae(Metschnikoff)],PPDA是菌落生长及产孢的最适培养基,大多数菌株对椰心叶甲有较强的致病力.选取强毒菌株MA4在田间进行防治效果的初步测定,结果表明,该菌株能显著降低椰心叶甲成虫的虫口密度.这些金龟子绿僵菌菌株是首次从海南的椰心叶甲僵虫中分离到的昆虫病原真菌,该菌对海南的椰心叶甲具有很好的生防潜能.%Coconut hispid beetles, Brontispa longissima (Gestro) (Coleoptera:Chrysomelidae) have been prevalent all over Hainan Island in China, and millions of coconut palms were attacked by this pest. The objective of this work was to isolate and identify strains of entomopathogenic fungi from natural infections of B. longissima collected from coconut palms.Conidia or mycelia collected from the surface of beetle cadavers were cultured and the colonies were screened on dodine oatmeal agar medium (DOA). The colony morphology, mycelia growth rate and conidial production were further studied on various agar media: PPDA, OMA, V8A and PDA. In order to confirm whether these isolates were pathogenic to B.longissima, their virulence to the beetles were

  8. Análise das deposições da pulverização aérea simulando a aplicação de Metarhizium anisopliae (Metsch na cultura da cana-de-açúcar Aerial spraying analysis for Metarhizium anisopliae (Metsch application in sugarcane crops

    Directory of Open Access Journals (Sweden)

    Hermes Geraldo Corrêa

    1992-01-01

    Full Text Available Aplicações de inseticida biológico constituído de esporos do fungo Metarhizium anisopliae (Metsch têm controlado satisfatoriamente a cigarrinha-da-cana, Mahanarva posticata (Stal, importante praga da cana-de-açúcar, em Alagoas. Este trabalho analisa as deposições da aplicação aérea na cultura da cana-de-açúcar, utilizando traçantes químicos e corantes. Foram determinados os depósitos do magnésio em fitas de papel acetinado colocadas transversalmente à linha de vôo e mediante análises de gotas recolhidas em cartões kromekote dispostos paralelamente às fitas. Nas condições em que as aplicações foram realizadas, verificou-se que nas faixas sobrepostas de 20 m, a recuperação foi de 44,5%,60% da qual se perde no solo de modo que, no final, apenas 18% do que foi aplicado efetivamente se deposita sobre a massa vegetal da cana. Portanto, se for realizada uma aplicação do entomopatógeno nas condições do ensaio, ela deverá apresentar uma deposição efetiva de aproximadamente 1/5 do total dos esporos aplicados por unidade de área.Biological control of spitthebug, Mahanarva posticata (Stal in sugarcane crop has been demonstrated possible and effective. Particular conditions in Northeastern of Brazil, where this pest causes severe damage, contribute to make aerial spraying suitable. Nevertheless, some good results in pest control are alternated with unsatisfactory ones. Some doubts on aerial spray application technology used have arisen, since technical information obtained in this Field are scarce. This trial was carried out in Alagoas State, Brazil, in 30-7-1985 with the aim of improving the efficiency of aerial application of the above biological pesticide mentioned. This paper presents some spray deposit patterns obtained with the Brazilian aircraft, namely IPANEMA EMB 201-A. The aerial spray application using this Brazilian aircraft and lane separations of 20 m, gave recovery of 44.5%, while 60% of it was lost in the

  9. Effect of Different Carbon Sources on the Growth of Metarhizium anisopliae IMI330189%不同碳源对金龟子绿僵菌IMI330189生长的影响

    Institute of Scientific and Technical Information of China (English)

    王磊; 赵霞; 曹广春; 张泽华

    2014-01-01

    The effect of different carbon sources on the fermentation of Metarhizium anisopliae IMI330189 was determined in this paper. The effects of six carbon sources-glucose, sucrose, soluble starch, fructose, glucose+fluctose (1:1) and the starch hydrolysate on the growth and the metabolism of M. anisopliae were discussed. The results showed that glucose+fluctose was the best carbon source among the six carbon sources, the biomass, growth rate and degradation time were 16.16 g/L, 0.4238 g/L·h, 48 h respectively, and the biomass were glucose+fluctose>sucrose>glucose>fructose>soluble starch. The starch hydrolysate were found to be better for M. anisopliae, the biomass of M. anisopliae was improved 14.3% compared with soluble starch. So it was demonstrated that the utilization of polysaccharide was insufficient for M. anisopliae and the addition of exogenous enzyme might improve the fermentation level and decrease the cost of M. anisopliae.%本文以单糖葡萄糖、果糖、蔗糖、葡萄糖+果糖(1:1)、多糖可溶性淀粉以及可溶性淀粉经酶处理后的水解产物为碳源,测定了发酵过程中糖消耗水平及绿僵菌生物量,分析了碳源对绿僵菌生长代谢的影响。结果表明,供试碳源中,葡萄糖+果糖的生物量最大(16.16 g/L)、达到最大生物量时的增长速度也最大(0.4238 g/L·h),并且被绿僵菌利用最快(48 h);其它依次为蔗糖、葡萄糖、果糖、可溶性淀粉;相同质量淀粉经外源α-淀粉酶水解12 h后作为碳源,与直接利用可溶性淀粉相比绿僵菌生物量增加14.3%左右。由此可见多糖基质碳源不利于绿僵菌的吸收利用,若在多糖基质碳源中添加外源糖类水解酶提前将多糖降解为单糖供绿僵菌生长,可提高绿僵菌对其利用率,进而提高发酵水平,降低发酵成本。

  10. The Influence of 3 Anti-fungal Agents on the Growth of Metarhizium and 6 Non-target Fungis%3种真菌抗生素对绿僵菌和6种非目标真菌的生长影响分析

    Institute of Scientific and Technical Information of China (English)

    杨华; 徐金柱; 秦长生; 揭育泽

    2014-01-01

    文章比较了两性霉素、伊曲康唑和放线菌酮对土壤内金龟子绿僵菌(Metarhizium anisopliae)、黄绿绿僵菌(M.flavoviride)和6种非目标真菌生长的影响。结果表明,伊曲康唑对毛霉(Mucor racemosus)和黑曲霉(Aspergillus niger)的抑菌效果较差,浓度达200μg/mL时对毛霉的抑制率仅达54.84%,且对绿僵菌生长的抑制作用显著大于两性霉素和放线菌酮(P<0.01),不适宜用于土壤绿僵菌的分离。两性霉素和放线菌酮在浓度为100μg/mL时,对6种非目标真菌抑制率达69%以上,可作为绿僵菌分离的真菌抗生素。%The effects of 3 anti- fungal agents on the growth of Metarhizium anisopliae,M.flavoviride strains and 6 non-target fungis were studied.The results showed that itraconazole did not have good inhibitory effects on Mucor racemosus and Aspergillus niger,the rate of M.racemosus controlling was 54.84%,but the inhibi-tory effects on the growth of Metarhizium were significantly higher than that of amphotericin and cycloheximide (P<0.01),suggesting it was not suitable for Metarhizium isolation from soil.Amphotericin and cycloheximide of 100μg/mL could inhibit over 69%of 6 non-target fungis,and were the most suitable anti-fungal agents for Metarhizium isolation.

  11. 绿僵菌田间流行及其与寄生蝇对蝗虫控制的调查%The Prevailing of Metarhizium anisopliae in Rangeland and Its Interaction with Parasitic Flies for Biological Control of Grasshoppers

    Institute of Scientific and Technical Information of China (English)

    董辉; 高松; 农向群; 丛斌; 张泽华; 孙大勇

    2011-01-01

    The grasshopper population showed two pathogenetic peaks after 20 d and 60 d of treatment by Metarhizium anisopliae conidia supplemented with oil-based suspension and bait. On the 20th d after treatment, the infectious rates with oil-based suspension and bait treatments rose to 54.5% and 34.79%, respectively, and difference between the treatments was significant. On the 60th d after treatment, the infected rates with both the treatments dropped to 23.62 % and 32.60%, respectively, and difference between treatments was also significant. The result indicated that the action of the oil-based suspension was faster than that of the bait, and the effect of the bait was more durative than that of the oil-based suspension. M. anisopliae might prevail naturally among the grasshopper population in this area. Infection of grasshoppers by M. anisopliae was independent on parasitic flies, suggesting that roles of biological control of grasshoppers by M. anisopliae and parasitic flies were additive.%在锡林浩特草场分别以6.0×107孢子·m-2施用量的绿僵菌Metarhizium anisopliae油悬浮剂、绿僵菌饵剂进行喷洒,调查绿僵菌在田间蝗虫中的流行、对寄生蝇寄生蝗虫的影响.结果表明,施用绿僵菌后,蝗虫种群分别于20d和60d出现2次感病高峰;处理后20d绿僵菌油悬浮剂和绿僵菌饵剂对蝗虫的感染率分别为54.55%和34.79%,绿僵菌油悬浮剂对蝗虫的感染率显著高于绿僵菌饵剂;处理后60d其感染率分别为23.62%和32.60%,绿僵菌饵剂对蝗虫的感染率显著高于绿僵菌油悬浮剂.说明绿僵菌油悬浮剂作用速度快于绿僵菌饵剂;绿僵菌饵剂持效性高于绿僵菌油悬浮剂;绿僵菌可以自然流行于该地区的蝗虫种群.绿僵菌孢子通过蝗虫病原地的二次流行,可向其周围环境扩散传播.表现出随着与绿僵菌饵剂处理区的距离增加,感染率逐渐降低;随着病原流行时间的延长,感病率逐渐升高.绿僵菌的

  12. 绿僵菌侵染后东亚飞蝗营养生理参数变化研究%Nutrition physiological parameter changes of Locusta migratoria manilensis ( Meyen ) infected with Metarhizium anisopliae

    Institute of Scientific and Technical Information of China (English)

    涂雄兵; 李志红; 牙森·沙力; 张泽华

    2011-01-01

    We investigated changes in the nutritional physiological parameters of Locusta migratoria manilensis (Meyen)infected with Metarhizium anisopliae at 18, 21, 24, 27 and 30℃.Total food consumption of 3rd -5th instar nymphs infected with M.anisopliae was 96.4, 108.3, 131.9, 103.2 and 249.5 mg respectively.The total food consumption of infected adults at 21, 24, 27 and 30℃, was 93.1, 121.8, 109.0 and 237.5 mg, respectively for males, and 108.7、195.5 、234.7、114.7 mg for females.The approximate digestive efficiency of infected nymphs decreased, and conversion efficiency increased, with increasing temperature, the latter being a negative value at lower temperatures.There were no significant differences in the approximate digestive efficiency of infected male adults, but that of infected female adults decreased after initially rising.The conversion efficiency of infected male adults decreased from 24 to 30℃, but that of infected female adults did not.The total food consumption of infected locusts was much less than that of CK locusts.We also studied changes in the food consumption of different developmental stages; the daily food consumption of CK nymphs increased at constant temperature and that of female adults was higher than that of male adults.Changes in the total food consumption of CK locusts were the same as changes in daily food consumption.However, there was no obvious relationship between these paramenters in locusts infected with M.anisopliae.%本文采用饵剂饲喂方法研究东亚飞蝗Locusta migratoria manilensis(Meyen)营养生理参数变化,结果表明:绿僵菌(Metarhizium anisopliae)侵染后,5个温度梯度下蝻期总取食量分别为96.4、108.3、131.9、103.2、249.5mg;21、24、27、30℃4个温度梯度下雄成虫总取食量为93.1、121.8、109.0、237.5 mg,雌成虫为108.7、195.5、234.7、114.7 mg.随温度升高,蝻期近似消化力降低,食物转化率升高,低温时转化率为负值.21~30℃范围内雄成虫近

  13. 绿僵菌产海藻糖水解酶培养条件研究%CULTURE CONDITIONS FOR THE PRODUCTION OF TREHALOSE-HYDROLYSING ENZYMES FROM METARHIZIUM ANISOPLIAE

    Institute of Scientific and Technical Information of China (English)

    李艳玲; 赵华; 王中康; 殷幼平; 彭国雄; 蔡绍皙; 夏玉先

    2004-01-01

    The filamentous fungus Metarhizium produces several extracellular disaccharide-degrading enzymes, including trehalose-hydrolysing enzymes. These enzymes play an important role in the antagonistic action of Metarhizium against most insects. The effects of different carbon sources and nitrogen sources on the production of trehalose-hydrolysing enzymes from Metarhizium anisopliae var. acridum isolate CQMaI02 were compared. The results indicated that CQMal02 could produce trehalose-hydrolysing enzymes in medium using glucose, maltose, sucrose, sorbitol or soluble starch as carbon source, but the best carbon source was soluble starch in terms of trehalose-hydrolysing activity and the number of isozymes produced, followed by sorbitol. Among the nitrogen sources tested, ammonium nitrogen and mixture of NaNO3 and organic nitrogen produced higher trehalose-hydrolysing activity than nitrate nitrogen.Comparison of trehalose-hydrolysing enzyme activity between the extracts of mycelia and the culture filtrate showed that the majority of trehalose-hydrolysing enzymes were secreted into liquid culture, only smaller amount of them were bound on cell wall.%丝状真菌绿僵菌能产生一系列二糖水解酶,其中包括海藻糖水解酶.这些酶在绿僵菌对昆虫的致病过程中起着重要的作用.本文研究了不同碳源、氮源对金龟子绿僵菌Metarhizium anisopliae var.acridum菌株CQMa102产生与分解昆虫血淋巴中海藻糖等二糖相关的海藻糖水解酶活性的影响.结果表明:分别以葡萄糖、麦芽糖、蔗糖、山梨醇和可溶性淀粉为碳源,金龟子绿僵菌均可产生海藻糖水解酶,但最佳碳源是可溶性淀粉,因为由其诱导产生的海藻糖水解酶具有最高的总活性和比活性以及更多的同工酶,山梨醇次之.硝态氮(NaNO3)作为唯一氮源时,几乎检测不出海藻糖水解酶活性,而铵态氮((NH4)2SO4)或NaNO3和有机氮(蛋白胨和酵母浸膏)混合氮源作氮源时,海藻糖水

  14. Pathogenicity and Efficacy of Different Metarhizium anisopliae and Beauveria bassiana and in Controlling Pantana phyllostachysae Chao Larvae%不同菌株绿僵菌和白僵菌对刚竹毒蛾幼虫致病力及林间防治试验

    Institute of Scientific and Technical Information of China (English)

    朱信华

    2013-01-01

    Pantana phyllostachysae Chao is an important kind of injurious pests of bamboo in Fujian Province. In order to screen entomopathogenic fungi strains with high virulence against P. phyllostachysae Larvae, the bioassay evaluation of one B. bassiana strain and five M. anisopliae strains against P. phyllostachysae Larvae was carried out through dipping in spore suspension. The results showed that the cumulative corrected mortality of P. phyllostachysae Larvae was over 63.16% with B. bassiana and M. anisopliae. The corrected mortality of P. phyllostachysae Larvae at 12 d after inoculated with MaFzzL-01 and Ma1291-2 was 90.93% and 84.21%, respectively. While LT50 values of two M. anisopliae strains, MaFzzL-01 and Ma1291-2, were at 5.97 d and 6.67 d, respectively. LC50 values of two strains, MaFzzL-01 and Ma1291-2, were 1.727×104 spore/mL and 2.209×104 spore/mL, respectively. The results of bioassay showed that MaFzzL-01 and Ma1291-2 strains had higher virulence to P. phyllostachysae Larvae than other strains did. When using 1.45×108 spore/mL of two M. anisopliae strains, Ma1291-2 and MaFzzL-01, to fight against larvae, good control efficiency in fields could be achieved.%刚竹毒蛾(Pantana phyllostachysae Chao)是福建省毛竹林的一种主要害虫,本研究应用5种金龟子绿僵菌(Metarhizium anisopliae)菌株和1种白僵菌(Beauveria bassiana)菌株,对3龄初幼虫进行感染力测定,旨在筛选出感染该虫的高致病力菌株。结果表明:应用1.45×107孢子/mL孢悬液感染12 d后,3龄初幼虫的校正死亡率均达到63.16%以上;其中MaFzzL-01和Ma1291-2两个绿僵菌株对该幼虫的致病力分别为90.93%和84.21%,其LT50值分别为5.97 d 和6.67 d, LC50值分别为1.727×104孢子/mL 和2.029×104孢子/mL。结果说明:绿僵菌MaFzzL-01和Ma1291-2两个菌株对该幼虫的致病力强,且致死速度快。应用1.45×108孢子/mL孢悬液进行林间防治可以取得良好效果。

  15. 金龟子绿僵菌深层培养产几丁质酶的研究%Production of Metarhizium anisopliae Chitinase in Submerged Culture and Study on the Producting Factors Influencing its Activity

    Institute of Scientific and Technical Information of China (English)

    杨革; 陈洪章; 李佐虎

    2005-01-01

    金龟子绿僵菌(Metarhizium anisopliae)是一种重要的昆虫寄生真菌,从自然罹病死亡的金龟子体内分离到一株金龟子绿僵菌(Metarhizium anisopliae)M-6.研究了该菌深层培养产几丁质酶的情况,几丁质酶合成的最佳碳源和诱导物为几丁质,在以0.5%(w/v)几丁质为碳源时,其产几丁质酶活达0.126IU·mL-1,在一定范围内增加培养基中几丁质浓度,葡萄糖浓度,微量元素盐浓度和碳氮比都能提高几丁质酶活.胆汁酸和吐温80作为表面活性剂能显著提高几丁质酶活.通过摇瓶试验得到优化培养基和培养条件.根据优化条件在摇瓶和3.7L发酵罐中分别进行产酶试验, 实验结果表明,几丁质酶活分别达到0.231 IU·mL-1和0.273 IU·mL-1.最适反应温度为55℃,最适反应pH6.0,在45℃,pH 3.0~ 9.5较为稳定.Zn2+、Ca2+、Ba2+和Mn2+离子对几丁质酶活性有明显的促进作用,而Hg2+、Co2+和Fe2+离子完全抑制几丁质酶的活性.

  16. Larvicidal and pupicidal properties of Acalypha alnifolia Klein ex Willd. (Euphorbiaceae) leaf extract and the microbial insecticide Metarhizium anisopliae (Metsch.) against lymphatic filarial vector, Culex quinquefasciatus..

    Science.gov (United States)

    This study was made to determine the mosquitocidal properties of Acalypha alnifolia leaf extract combined with the use of Metarizhium anisopliae spores for control of the lymphatic filariasis vector Culex quinquefasciatus. The methanolic leaf extract showed larvicidal and pupicidal effects after 24...

  17. Larvicidal and pupicidal properties of Acalypha alnifolia Klein ex willd.(Euphorbiaceae) leaf extract and the microbial insecticide Metarhizium anisopliae (Metsch.) against lymphatic filarial vector, Culex quinquefasciatus Sa

    Science.gov (United States)

    This study was made to determine the mosquitocidal properties of Acalypha alnifolia leaf extract combined with the use of Metarizhium anisopliae spores for control of the lymphatic filariasis vector Culex quinquefasciatus. The methanolic leaf extract showed larvicidal and pupicidal effects after 24...

  18. Identification of a hybrid PKS-NRPS required for the biosynthesis of NG-391 and NG-393 metabolites in Metarhizium anisopliae

    Science.gov (United States)

    A 19,818 kb genomic region harboring six predicted ORFs was identified in M. anisopliae ARSEF 2575. The ORF4 CDS, putatively encoding a hybrid polyketide synthase/nonribosomal peptide synthetase (PKS-NRPS) was targeted using Agrobacterium-mediated gene knockout. Homologous, but not heterolog...

  19. 绿僵菌大孢种的原生质体及其染色体组型研究%Studies on the Protoplasts and Karyotype of Metarhizium anisopliae var. majus, a Entomogenous Fungus

    Institute of Scientific and Technical Information of China (English)

    时连根

    2000-01-01

    研究了绿僵菌大孢种(Metarhizium anisopliae var.majus)原生质体的分离制备条件及其性状,并用脉冲凝胶电泳(PFGE)技术分析了其染色体组型.以6 mg/mL Driselase、6.0 mg/mL Novozyme234、6 mg/mL Zymolyase混合液中添加3mg/mL牛血清白蛋白作为酶液,以含0.05 mol/L EDTA的0.7 mol/mL NaCl溶液(pH 5.8)为渗透压稳定剂,在30℃下轻轻振荡处理幼嫩菌丝1.5 h,为绿僵菌大孢种原生质体分离的适宜条件.观察表明:原生质体的无核率为25.3 %,有核率为74.7 %,其中单核率为53.6 %.绿僵菌大孢种至少具有7条染色体,估算大小为1.1~6.5 Mb,3菌株染色体组型大小平均为24.4 Mb.

  20. Study on Production Conditions and Properties of Chitinase by Metarhizium anisopliae Ma83%绿僵菌Ma83固态发酵几丁质酶和部分酶学性质的初步研究

    Institute of Scientific and Technical Information of China (English)

    蔡荟梅; 刘斌; 蔡敬民; 吴克; 樊美珍

    2003-01-01

    研究了金龟子绿僵菌Metarhizium anisopliae Ma83菌株产几丁质酶的固态发酵条件和粗酶液的酶学性质.研究结果显示,当以麸皮∶蚕蛹粉为3∶1作为培养基,最适氮源为1% NaNO3,起始pH为6.5,发酵温度为31℃,接种量为2mL液态种子时酶活力最高.此外,添加稻壳对Ma83产几丁质酶有促进作用.该菌株在生长2d时,酶活力达33.9U/g干培养基.酶的最适反应温度为50℃,最适反应pH为6.0;不同温度保温1h后,酶的半失活温度为42 ℃.不同pH值的缓冲溶液中(25℃)放置1h后,酶在pH 5.0~6.0条件下稳定性最高.

  1. Cellular Defence Reaction of American Cockroach,Periplaneta americana to Metarhizium anisopliae Isolate CQMa102%美洲大蠊血细胞对金龟子绿僵菌CQMa102菌株的免疫反应

    Institute of Scientific and Technical Information of China (English)

    钟伟; 殷幼平

    2003-01-01

    将蜚蠊的非致病菌--金龟子绿僵菌(Metarhizium anisopliae)CQMa102菌株的分生孢子注入美洲大蠊(Periplaneta americana)血腔后,诱导了蜚蠊血淋巴强烈的免疫反应,其血细胞数量发生了显著变化,血细胞通过吞噬作用及形成结节使绿僵菌孢子钝化,萌发受到抑制,或被彻底分解;注射高浓度孢子时,蜚蠊的免疫体系受到严重破坏,容易感染细菌而死亡;但注射低浓度孢子却能够增强蜚蠊的免疫机能.生理盐液也能诱导蜚蠊血淋巴的免疫反应.

  2. Effect of Metarhizium anisopliae on Insect Pests in Lawn%绿僵菌与化学农药对草坪地下害虫控制效果的比较

    Institute of Scientific and Technical Information of China (English)

    李有志; 马骏; 黎家文; 文礼章

    2005-01-01

    近年来,随着城市绿化草坪和高尔夫球场总面积的扩大,影响草坪质量的虫害问题也日益突出。由于农药在草坪上的大量使用,其污染和残留问题已引起人们的重视。寻找污染少、残留低,又能有效地控制草坪害虫的杀虫剂是当前草坪害虫治理中的课题之一。金龟子绿僵菌(Metarhizium anisopliae)作为一种生物农药已被广泛地用于防治蝗虫、蟑螂等害虫。作者就绿僵菌和两种化学药剂对草坪害虫的防治效果作了比较试验,现报道如下。

  3. Antagonism of entomopathogenic fungi by Bacillus spp. associated with the integument of cicadellids and delphacids.

    Science.gov (United States)

    Toledo, Andrea; López, Silvina; Aulicino, Mónica; de Remes-Lenicov, Ana María; Balatti, Pedro

    2015-06-01

    Entomopathogenic fungi are potential tools to biocontrol cicadellids and delphacids, two groups of insects that cause extensive damage to agricultural crops. However, bacteria living on the host cuticle may inhibit fungal growth. In the present work, following the molecular characterization of 10 strains of Bacillus isolated from the integument of cicadellids and delphacids, we selected isolates of the fungi Beauveria bassiana and Metarhizium anisopliae that are resistant to the antimicrobials secreted by these bacterial strains. The antagonistic activity of the 10 bacterial isolates belonging to the genus Bacillus (i.e., B. amyloliquefaciens, B. pumilus, and B. subtilis) against 41 isolates of Bea. bassiana and 20 isolates of M. anisopliae was investigated in vitro on tryptic soy agar using the central disk test. With this approach, isolates of Bea. bassiana and M. anisopliae resistant to antagonistic bacteria were identified that can be further developed as biological control agents.

  4. 应用白僵菌和绿僵菌防治浙江双栉蝠蛾试验%Experiment of Beauveria bassiana and Metarhizium anisopliae on controlling Bipectilus zhejiangensis

    Institute of Scientific and Technical Information of China (English)

    童应华; 谢家冬; 祝德平

    2012-01-01

    Highly pathogenic Beauveria bassiana strain B3 and Metarhizium anisopliae strain Ma1291-2 were used for controlling Bipectilus zhejiangensis with different release methods. The results of 2-year controlling showed that effects of root irrigation was better than that of spreading with 2 strains, otherwise, the best effect was using 2 methods contemporarily (spreading + irrigation) in different time ( root irrigation in mid-March after spread in early March). There were no significant differences of the degraded shoot rate controlled between Beauveria bassiana and M. anisopliae in the first year, but the degraded shoot rate being controlled with strain Mal291-2 was lower significantly in the second year. By root spreading + irrigation with B3, the effects were 72.20% and 36.76% respectively in the first year and second year, and that of Mal291-2 was 79.28% and 48.70% respectively. Therefore, the 2 strains have application value for controlling Bipectilus zhejiangensis in forests, and the effect of application by root spreading + irrigation root was better in different time.%应用高致病力球孢白僵菌[ Beauveria bassiana( Bals.)Vuil] B3和金龟子绿僵菌[Metarhizium anisopliae( Metsch.)Sorokin] Ma1291-2菌株,以不同的施菌方式防治浙江双栉蝠蛾(Bipectilus zhejiangensis Wang),统计分析2a的防治效果.结果表明,2菌株均体现笋基灌施的防治效果较菌土撤施好,但以3月上旬进行撒施,3月中旬进行灌施(撒施+灌施)的防治效果最好.2菌株防治后第1年的退笋率无显著差异,而绿僵菌防治后第2年退笋率显著较低.以撒施+灌施防治后第1年和第2年,白僵菌B3菌株的防治效果分别为72.20%和36.76%,绿僵菌Ma1291-2菌株的防治效果分别为79.28%和48.70%.因此,2菌株均有林间应用价值,并以不同时间撒施+灌施的施菌方式防治,效果更佳.

  5. Growth and sporulation of Metarhizium flavoviride var. Flavoviride on culture media and lighting regimes

    Directory of Open Access Journals (Sweden)

    Onofre Sideney Becker

    2001-01-01

    Full Text Available Entomopathogenic fungi from the genus Metarhizium are largely used for the biological control of agricultural pests by conidia spreading on the field. Although conidia production is well studied in M. anisopliae, only few research studies were done in M. flavoviride. The present work was carried out alming to evaluate the Mycelial growth and sporulation of the entomopathogenic fungus Metarhizium flavoviride var. flavoviride growing at 27 ± 2°C on Potato-dextrose-agar (PDA, Czapek-agar (CZP and a complete agar medium (CM under three lighting regimes, (continuous illumination, light/dark cycle and an black light/dark cycle were investigated. A completely randomized 3 × 3 (culture media × lighting regime factorial design with four replicates was used. The best mycelial growth and sporulation occurred on the PDA and CM media under continuous illumination (P <= 0,05.

  6. MALDI-TOF mass spectrometry applied to identifying species of insect-pathogenic fungi from the Metarhizium anisopliae complex

    Science.gov (United States)

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has proven to be a powerful tool for taxonomic resolution of microorganisms. In this proof-of-concept study, we assessed the effectiveness of this technique to track the current gene sequence-based phylogenet...

  7. Cultivation of entomopathogenic fungi for the search of antibacterial compounds.

    Science.gov (United States)

    Lee, Si-Young; Nakajima, Ikuo; Ihara, Fumio; Kinoshita, Hiroshi; Nihira, Takuya

    2005-11-01

    Entomopathogenic fungi are a rich source of natural bioactive compounds. To establish cultivation conditions which facilitate the production of bioactive compounds and to select good genera among entomopathogenic fungi as the producer, 47 typical entomopathogenic fungi were tested for their ability to produce antibiotic activity. Thirty-eight strains (81%) and 30 strains (64%) of these fungi produced either anti-Bacillus compounds or anti-Staphylococcus compounds, respectively, indicating that the majority of the entomopathogenic fungi tested possessed the ability to produce antibacterial compounds. Using 9 representative strains (Aschersonia sp. HF724, Beauveria bassiana HF338, Cordyceps ramosopulvinata HF746, Metarhizium anisopliae HF293, Metarhizium flavoviride HF698, Nomuraea rileyi HF588, Paecilomyces fumosoroseus HF254, Paecilomyces tenuipes HF419, and Verticillium lecanii HF238), the cultivation conditions in liquid medium were surveyed with respect to the cultivation procedure and medium composition, particularly in terms of the presence or absence of insect-derived materials. At 26 degrees C, M. anisopliae HF293, N. rileyi HF588, and V. lecanii HF238 strains produced clear antibiotic activity against Bacillus and Saccharomyces, but only in the presence of insect-derived materials, suggesting that the production of antibacterial/antifungal compounds by entomopathogenic fungi is triggered by the presence of insect-derived materials.

  8. 绿僵菌治蝗对草原昆虫生物多样性的影响%The Effect on Grassland Insects Biodiversity of Metarhizium anisopliae Controlling Grasshopper

    Institute of Scientific and Technical Information of China (English)

    董辉; 张泽华; 高松; 农向群; 丛斌

    2011-01-01

    Metarhizium anisopliae to control locust plagues is a sustainable low-cost biological control measure with no pollution effects on society, and has an important significance in the protection of grassland ecosystem biodiversity. Through investigation on of insect community composition and structural diversity in different grassland locust control areas in Xilinguolemeng of Inner Mongolia, the influence on biodiversity was exposed. Major dominant pests and potential secondary pests were analyzed, which provides a basis for reference for establishing a grassland locust integrated control system.%绿僵菌治蝗是一项低耗、无公害、可持续的生物治理蝗害措施,对保护草原生态系统生物多样性具有重要意义.通过对内蒙古锡林郭勒盟草原蝗虫不同防治区昆虫群落的组成和结构多样性调查与研究,揭示了化学农药对生物多样性的负面影响;评价了绿僵菌制剂在保护和调节生物多样性方面的作用;明确了目前的主要优势种害虫和可能上升为主要害虫的次要害虫,对建立草原蝗虫的综合治理和其他害虫控制提供参考依据.

  9. Abundance, Genetic Diversity and Persistence of Metarhizium Spp. Fungi from Soil of Strawberry Crops and Their Potential as Biological Control Agents against the Two-Spotted Spider Mite Tetranychus urticae

    DEFF Research Database (Denmark)

    Castro, Thiago Rodriguesde

    The growing demand for strawberries has imposed challenges, especially regarding the control of pests. Many farmers report problems with reduced chemical control efficiency, probably due to selection of resistant populations of insects and mites. An alternative is the use of biological control...... using pathogenic fungi as a tool in integrated pest management. Metarhizium spp. (Hypocreales: Clavicipitaceae) are generalist entomopathogenic fungi with worldwide distribution and can cause diseases in a large number of hosts. Many studies on the development of Metarhizium as a biological control...... agent were performed, but this bulk of knowledge is in remarkable contrast to the lack of research on the fundamental ecology of Metarhizium in agroecosystems. This thesis aimed to evaluate the establishment, persistence and dispersal of these entomopathogenic fungi in strawberry crop soil...

  10. Effects of the Secondary Metabolites of Metarhizium anisopliae on Newcastle Disease Vaccine%苦马豆素对新城疫疫苗免疫效果的影响

    Institute of Scientific and Technical Information of China (English)

    吴延磊; 杨鸣琦; 商育锦; 施珊珊; 吴小胜

    2011-01-01

    研究金龟子绿僵菌次级代谢产物对鸡新城疫疫苗免疫效果的影响.将100只14日龄健康无免疫雏公鸡随机分为5组,第Ⅰ~Ⅲ组分别注射高、中、低(4、2、1 mg/kg)3种剂量的金龟子绿僵菌次级代谢产物;Ⅳ组和Ⅴ组分别为无佐剂组对照组(No Adjuvant,NA)和空白对照组.第Ⅰ~Ⅳ组动物于14日龄以新城疫病毒(NDV) LaSota株弱毒活疫苗滴鼻进行第1次免疫,2周后加强免疫1次.分别于首免前和首免后连续6周在各免疫组随机抽取10只鸡检测血清中ND-HI抗体效价;于每次免疫后24 h检测外周血中白细胞数;于免疫后42日龄检测免疫器官指数;用MTT法检测脾脏淋巴细胞的增殖活性.金龟子绿僵菌次级代谢产物剂量为1~2mg/kg,大多数试验组鸡的抗体效价与雏鸡脏器(胸腺,脾脏,法氏囊)指数及外周血中白细胞数都显著高于对照组.金龟子次级代谢产物单独或其协同脂多糖(LPS),植物血凝素P(PHA-P)刺激时,鸡脾脏淋巴细胞的增殖活性增强.金龟子绿僵菌次级代谢产物剂量为4mg/kg时,试验组鸡的抗体效价、免疫器官指数和外周血中的白细胞数均减少;鸡脾脏淋巴细胞的增殖活性降低.%The trial was to study the effect of the secondary metabolites of Metarhizium anisopliae on newcastle disease vaccine. 100 14-day-old chickens were randomly assigned into equal five groups. The chickens in three experimental groups were intramuscularly injected respectively with secondary metabolites of Metsrhizium anisopliae at high, medium, and low dose and vaccinated with Newcastle disease vaccine. Two control groups were administrated with no adjuvant (NA) and normal group. Four groups (group Ⅰ to Ⅳ) were immunized with the LaSota live vaccine against Newcastle disease virus(NDV)for two times with 14 days interval. In continuous 6 weeks before and after vaccination, 10 chickens were sampled randomly from each group for the determination of

  11. Entomopathogenic fungi in cornfields and their potential to manage larval western corn rootworm Diabrotica virgifera virgifera.

    Science.gov (United States)

    Rudeen, Melissa L; Jaronski, Stefan T; Petzold-Maxwell, Jennifer L; Gassmann, Aaron J

    2013-11-01

    Entomopathogenic ascomycete fungi are ubiquitous in soil and on phylloplanes, and are important natural enemies of many soil-borne arthropods including larval western corn rootworm, Diabrotica virgifera virgifera, which is a major pest of corn. We measured the prevalence of Beauveria bassiana and Metarhizium anisopliae sensu lato in ten cornfields in Iowa, USA by baiting with larval insects. B. bassiana and M. anisopliae s.l. were present in 60% ± 6.3% and 55% ± 6.4% of soil samples, respectively. Subsequent laboratory bioassays found that some M. anisopliae s.l. strains collected from cornfields killed a greater proportion of D.v. virgifera larvae than a standard commercial strain.

  12. REVESTIMENTO DE PANOS COM POLÍMERO AUMENTA A EFICIÊNCIA DO FUNGO Metarhizium anisopliae NO CONTROLE DE Aedes aegypti

    Directory of Open Access Journals (Sweden)

    Aline Teixeira Carolino

    2017-04-01

    Full Text Available A estratégia de aplicação do fungo sobre panos pretos é uma ferramenta eficiente no controle do mosquito A. aegypti. Para aprimorar a técnica e aumentar o número de conídios disponíveis na superfície do pano, os panos de algodão preto foram revestidos com alginato de sódio (ALG e impregnados com o fungo entomopatogênico M. anisopliae (F. O fungo foi pulverizado sobre os panos e após secos, ficaram em contato com os mosquitos por 48h. Nos ensaios de germinação, foi possível verificar que o alginato não influenciou na viabilidade do fungo. No meio de cultura SDA acrescido com alginato foi verificado germinação de 97% dos conídios e 98% no grupo controle, sem adição do polímero. Nos ensaios de sobrevivência, o tratamento ALG+F foi mais eficiente em reduzir a sobrevivência dos mosquitos sendo verificada 12% de sobrevivência e S50 de 3 dias. O tratamento Fungo+Tween apresentou menor eficiência, sendo constatado 42% de sobrevivência e S50 de 7 dias.

  13. Virulence of Metarhizium anisopliae var. anisopliae to the Larvae of Melanotus cribricollis (Coleoptera: Elateridae)%金龟子绿僵菌小孢变种对筛胸梳爪叩甲幼虫致病力的生物测定

    Institute of Scientific and Technical Information of China (English)

    王鹏; 张亚波; 舒金平; 邓顺; 王浩杰

    2010-01-01

    从筛胸梳爪叩甲Melanotus cribricollis僵死幼虫体内分离得到1株金龟子绿僵菌小孢变种Metarhizium anisopliae var.anisopliae菌株WP,首次采用拌土法对该菌寄生筛胸梳爪叩甲幼虫的致病力进行测定,并分析外界条件对绿僵菌致病力的影响.在WP菌株1×104~1×107孢子/g干土7个孢子浓度梯度下,随着浓度的提高,筛胸梳爪叩甲的校正死亡率从20%上升至100%.在15、20、25、30℃4个温度下,该菌对筛胸梳爪叩甲的致死率随着温度升高而上升.30℃时,筛胸梳爪叩甲死亡率达96.7%;当降至15℃,筛胸梳爪叩甲不会被寄生.在6%~18%土壤湿度条件下,土壤湿度的升高会提高WP菌株对筛胸梳爪叩甲的致死率,6%的土壤湿度不会导致该虫染病.

  14. Genetic diversity and pathogenicity of Metarhizium spp. associated with the white grub Phyllophaga capillata (Blanchard) (Coleoptera: Melolonthidae) in a soybean field.

    Science.gov (United States)

    Lopes, R B; Souza, D A; Oliveira, C M; Faria, M

    2013-08-01

    Phyllophaga capillata (Blanchard) was recently described as a soybean pest in Brazil. The occurrence of Metarhizium spp. associated with adults and their pathogenicity were investigated. Natural prevalence of Metarhizium anisopliae was 0.51% in adults collected by light trap. Most Metarhizium isolates were identified as Metarhizium robertsii, although M. anisopliae sensu stricto and a few unidentified isolates were also found. Bioassays with representative isolates selected from different genetic groups resulted in ≤5.6% confirmed mortality against larvae and adults, suggesting low potential of microbial control of this pest by use of indigenous Metarhizium isolates.

  15. 金龟子绿僵菌附着胞分化及其与环腺苷酸cAMP的关联性研究%Appressorial differentiation and its association with cAMP in the insect pathogenic fungus Metarhizium anisopliae

    Institute of Scientific and Technical Information of China (English)

    段志兵; 高强; 吕丁丁; 石少华; BUTT Tariq M.; 王成树

    2009-01-01

    Host recognition and appressorium differentiation are the pivotal steps for insect pathogenic fungi to initiate infection process. In this study, appressorial differentiation was studied by comparing a mutant, which has been identified with the loss of genetic materials, with the wild-type strain of the insect pathogenic fungus Metarhizium anisopliae. The observations showed that both the mutant and wild-type strain could not only produce appressoria on the tips of newly germinated spores, but also form multiple infection structures from the terminals of branching mycelia on insect cuticle. In contrast to the wild-type, the frequency of appressorial formation was significantly reduced and in addition, no clear mucilaginous sheath was produced by the mutant appressoria. The study shows that the cuticle degrading enzyme subtilisin is not involved in appressorial differentiation, orindispensable in cuticle digestion. A significantly low frequency of appressorial differentiation by the mutant was coincided with its lower intracellular cAMP level in comparison to the wild-type. Addition of exogenous cAMP could significantly increase the frequency of appressorial differentiation by the mutant, indicating that cAMP signaling pathway is potentially involved in regulation of appressorial differentiation in M. anisopliae.%寄主识别与附着胞分化是虫生真菌启动侵染过程的首要步骤.本文利用先前获得的金龟子绿僵菌基因缺失突变株与其野生型一起进行附着胞分化研究.接种后不同时间下的观察表明,绿僵菌突变株或野生型的附着胞既可以在萌发不久的芽管顶端形成,也可以在伸长菌丝分支的顶端形成.与野生型不同的足,突变株附着胞的分化频率显著下降,附着胞周围也缺乏粘液层的产生.研究表明,绿僵菌的类枯草杆菌类体壁降解酶对于附着胞分化不产生影响,对体壁降解也非完全必需的.与突变株附着胞分化频率显著降低相

  16. Screening of Metarhizium spp. strains for anticancer indolizidine alkaloid production and its rapid detection by MS analysis Seleção de linhagens de Metarhizium spp. para a produção de alcalóide indolizidínico anticancerígeno e sua rápida detecção por MS análise

    Directory of Open Access Journals (Sweden)

    Oselys Rodriguez Justo

    2007-12-01

    Full Text Available Six fungi strains (M. anisopliae 3935, 4516, 4819, PL57, PL43 and M. flavoviride CG291 were studied regarding their ability to produce an anticancer indolizidine alkaloid. The culture process was carried out in Shaken flask at 26ºC and 200 rpm using three different culture medium containing oat meal extract supplemented with glucose and DL-lysine or Czapek culture medium. The mycelial extracts produced by Metarhizium spp. cultures were directly submitted to electrospray ionization mass spectrometry (ESI-MS analysis and the highest alkaloid concentration (approximately, 6 mg.L-1 was reached when M. anisopliae 3935 was tested.O presente trabalho teve como objetivo avaliar diferentes cepas dos fungos M. anisopliae e M. flavoviride ao respeito da sua capacidade de produzir um alcalóide anticancerígeno, por fermentação em frascos erlenmeyers usando três meios de cultura distintos. De seis cepas testadas, quatro foram capazes de produzir o composto de interesse, M. anisopliae 3935, PL57 e PL43 e M. flavoviride CG291, sendo que a maior concentração de alcalóide (aproximadamente, 6 mg.L-1 foi produzida pelo M. anisopliae 3935, contendo um meio constituído de extrato de farinha de aveia, glicose e DL-lisina a 26ºC e 200 rpm.

  17. [Natural ocurrence of entomopathogenic fungi in soils cultivated with Paraguay tea (Ilex paraguariensis St. Hil.) in Misiones, Argentina].

    Science.gov (United States)

    Schapovaloff, María E; Angeli Alves, Luis F; Urrutia, María I; López Lastra, Claudia C

    2015-01-01

    This study aimed to morphologically isolate, identify and characterize entomopathogenic fungi present in soils cultivated with Paraguay tea (Ilex paraguariensis). A survey of native entomopathogenic fungi was conducted from 40 soil samples grown with Paraguay tea in the province of Misiones, Argentina, from May 2008 to June 2010. The soil dilution plate methodology on selective culture media was used to isolate microorganisms. Taxonomic identification was performed using macroscopic and microscopic characters and specific keys. Twenty nine strains, belonging to the species Beauveria bassiana (n = 17), Metarhizium anisopliae (n = 2) and Purpureocillium lilacinum (n = 10) were isolated and identified.

  18. Association between entomopathogenic nematodes and fungi for control of Rhipicephalus microplus (Acari: Ixodidae).

    Science.gov (United States)

    Monteiro, Caio Márcio Oliveira; Araújo, Laryssa Xavier; Matos, Renata Silva; da Silva Golo, Patrícia; Angelo, Isabele Costa; de Souza Perinotto, Wendell Marcelo; Coelho Rodrigues, Camila Aparecida; Furlong, John; Bittencourt, Vânia Rita Elias Pinheiro; Prata, Márcia Cristina Azevedo

    2013-10-01

    The aim of the study was to assess the effect of the association of entomopathogenic nematodes and fungi on Rhipicephalus microplus. The nematodes used were Heterorhabditis bacteriophora HP88 and Heterorhabditis indica LPP1 and the fungi were Metarhizium anisopliae IBCB 116 and Beauveria bassiana ESALQ 986. In the groups treated with the fungi, the females were immersed for 3 min in a conidial suspension, while in the groups treated with the nematodes, the ticks were exposed to infective juveniles. To evaluate the interaction between entomopathogens, the females were first immersed in a conidial suspension and then exposed to the nematodes. The egg mass weight and hatching percentage values of the groups treated with M. anisopliae IBCB 116 and B. bassiana ESALQ 986 in the two experiments were statistically similar (p > 0.05) to the values of the control group. In the groups treated only with nematodes, there was a significant reduction (p fungi, there was a significant reduction (p fungi alone varied from 31 to 55%. In the groups treated with nematodes associated or not with fungi, the control percentage was always greater than 90% and reached 100% in the group treated with H. bacteriophora HP88 associated with the fungus M. anisopliae IBCB 116.

  19. Abundance, Genetic Diversity and Persistence of Metarhizium Spp. Fungi from Soil of Strawberry Crops and Their Potential as Biological Control Agents against the Two-Spotted Spider Mite Tetranychus urticae

    DEFF Research Database (Denmark)

    Castro, Thiago Rodriguesde

    The growing demand for strawberries has imposed challenges, especially regarding the control of pests. Many farmers report problems with reduced chemical control efficiency, probably due to selection of resistant populations of insects and mites. An alternative is the use of biological control...... agent were performed, but this bulk of knowledge is in remarkable contrast to the lack of research on the fundamental ecology of Metarhizium in agroecosystems. This thesis aimed to evaluate the establishment, persistence and dispersal of these entomopathogenic fungi in strawberry crop soil...... in Inconfidentes, Minas Gerais, Brazil; and to study the diversity and abundance of species of Metarhizium isolated from organic and conventional strawberry crop soils, and from the field margins in Brazil and Denmark. The effectiveness of new species of Metarhizium recently found in Brazil, was evaluated against...

  20. Effect of certain entomopathogenic fungi on oxidative stress and mortality of Periplaneta americana.

    Science.gov (United States)

    Chaurasia, Abhilasha; Lone, Yaqoob; Wani, Owais; Gupta, U S

    2016-02-01

    The present paper reports the effects of Metarhizium anisopliae, Isaria fumosoroseus and Hirsutella thompsonaii on Periplaneta americana. I. fumosoroseus and H. thompsonaii were cultured at 28±1°C on potato carrot agar and M. anisopliae was cultured at 28±1°C on potato dextrose agar for 14days. Conidial suspensions of fungi were given to cockroaches through different routes. M. anisopliae shows high virulence against adult cockroaches and mortality ranges from 38.65% to 78.36% after 48h. I. fumosoroseus and H. thompsonii show less virulence compared to M. anisopliae. We also investigated the effect of these three fungi on the activity of lactate dehydrogenase, lipid peroxidation and catalase in different tissues of the insect to gain an understanding of the different target site. The result suggested that the activity of lactate dehydrogenase, catalase and level of malondialdehyde varies in different organs and through different routes of exposure. Based on mortality percentages, all tested fungi had high potentials for biocontrol agents against P. americana. Our study reveals for the first time that I. fumosoroseus and H. thompsonaii fungal infections initiate oxidative stress in the midgut, fat body, whole body and hemolymph of cockroach thereby suggesting them to be the target organs for oxidative damage.

  1. Unveiling the oxidative metabolism of Rhipicephalus microplus (Acari: Ixodidae) experimentally exposed to entomopathogenic fungi.

    Science.gov (United States)

    Tunholi-Alves, Vinícius Menezes; Tunholi Alves, Victor Menezes; da Silva, Jairo Pinheiro; Nora Castro, Rosane; Salgueiro, Fernanda Barbosa; Perinotto, Wendell Marcelo de Souza; Gôlo, Patrícia Silva; Camargo, Mariana Guedes; Angelo, Isabele da Costa; Bittencourt, Vânia Rita Elias Pinheiro

    2016-10-01

    Rhipicephalus microplus is an important tick in tropical regions due to the high economic losses caused by its parasitism. Metarhizium anisopliae and Beauveria bassiana are well-known entomopathogenic fungi that can afflict R. microplus ticks. The development of new targets and strategies to control this parasite can be driven by studies of this tick's physiology. Recently, it was reported that when exposed to adverse physiological conditions, ticks can activate fermentative pathways, indicating transition from aerobic to anaerobic metabolism. Nevertheless, the precise mechanism by which entomopathogenic fungi influence R. microplus metabolism has not been clarified, limiting understanding of the tick-fungus association. Thus, the present study aimed to evaluate the effect of infection of ticks by M. anisopliae and B. bassiana on the amount of selected carboxylic acids present in the hemolymph, enabling increased understanding of changes previously reported. The results showed preservation in the concentrations of oxalic, lactic, and pyruvic acids in the hemolymph 24 and 48 h after dropping from cattle; while there were variations in the concentration of these carboxylic acids after infection of female ticks to M. anisopliae and B. bassiana. Significant increases were observed in the concentration of oxalic and lactic acids and significant reduction of pyruvic acid for both observation times (24 and 48 h) after infection by entomopathogenic fungi. These results indicate that B. bassiana and M. anisopliae infection alters the basal metabolism of R. microplus females, resulting in the activation of fermentative pathways.

  2. Microbial control of the invasive spiraling whitefly on cassava with entomopathogenic fungi.

    Science.gov (United States)

    Boopathi, Thangavel; Karuppuchamy, Palaniappan; Singh, Soibam B; Kalyanasundaram, Manickavasagam; Mohankumar, S; Ravi, Madhaiyan

    2015-01-01

    The entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, Lecanicillium lecanii and Isaria fumosorosea were tested for their efficacy in managing the exotic spiraling whitefly Aleurodicus dispersus (Hemiptera, Aleyrodidae) on cassava (Manihot esculenta) during 2 seasons (2011-2012 and 2012-2013). The fungi I. fumosorosea and L. lecanii exhibited promising levels of control (> 70% mortality of the A. dispersus population). The percent mortality increased over time in both seasons. Application of I. fumosorosea was highly pathogenic to A. dispersus in both seasons compared to the other entomopathogenic fungi. Analysis of the percent mortality in both seasons revealed differences in efficacy between 3 and 15 days after treatment. The season also influenced the effects of the fungi on the A. dispersus population. Thus, entomopathogenic fungi have the potential to manage A. dispersus infestation of cassava.

  3. OCCURRENCE OF ENTOMOPATHOGENIC FUNGI IN SOILS FROM FESTUCA PRATENSIS HUDS. CROP

    Directory of Open Access Journals (Sweden)

    Roman Kolczarek

    2014-04-01

    Full Text Available Entomopathogenic fungi are the largest group of microorganisms existing in the soil environment. Occurrence and pathogenicity of entomopathogenic fungi in soil is dependent on many factors affecting the soil environment. The aim of this study was to compare the species composition and the intensity of the occurrence of entomopathogenic fungi in soils from monoculture crops of Festuca pratensis Huds. The study material consisted of soil samples taken from the experiment conducted in two experimental stations of the Research Centre for Cultivars Study. The insecticides fungi were isolated from soil using a method of the selective substrate. Three species of entomopathogenic fungi Beauveria bassiana, Isaria fumosorosea and Metarhizium anisopliae were isolated from the study soils using the selective medium.

  4. Biolarvicidal and pupicidal activity of Acalypha alnifolia Klein ex Willd.(Family:Euphorbiaceae) leaf extract and microbial insecticide, Metarhizium anisopliae(Metsch.)against malaria fever mosquito Anopheles stephensi Liston

    Science.gov (United States)

    This study was made to determine the biological activity of Acalypha alnifolia leaf extract and the microbial insecticide Metarizhium anisopliae against larvae and pupae of the malaria vector Anopheles stephensi. Ethanolic A. alnifolia leaf extract tested against 1st through 4th instars and pupae o...

  5. Characterization of Metarhizium species and varieties based on molecular analysis, heat tolerance and cold activity

    Science.gov (United States)

    Fernandes, E.K.K.; Keyser, C.A.; Chong, J.P.; Rangel, D.E.N.; Miller, M.P.; Roberts, D.W.

    2010-01-01

    Aims: The genetic relationships and conidial tolerances to high and low temperatures were determined for isolates of several Metarhizium species and varieties. Methods and Results: Molecular-based techniques [AFLP and rDNA (ITS1, ITS2 and 5??8S) gene sequencing] were used to characterize morphologically identified Metarhizium spp. isolates from a wide range of sources. Conidial suspensions of isolates were exposed to wet heat (45 ?? 0??2??C) and plated on potato dextrose agar plus yeast extract (PDAY) medium. After 8-h exposure, the isolates divided clearly into two groups: (i) all isolates of Metarhizium anisopliae var. anisopliae (Ma-an) and Metarhizium from the flavoviride complex (Mf) had virtually zero conidial relative germination (RG), (ii) Metarhizium anisopliae var. acridum (Ma-ac) isolates demonstrated high heat tolerance (c. 70-100% RG). Conidial suspensions also were plated on PDAY and incubated at 5??C for 15 days, during which time RGs for Ma-an and Ma-ac isolates were virtually zero, whereas the two Mf were highly cold active (100% RG). Conclusions: Heat and cold exposures can be used as rapid tools to tentatively identify some important Metarhizium species and varieties. Significance and Impact of the Study: Identification of Metarhizium spp. currently relies primarily on DNA-based methods; we suggest a simple temperature-based screen to quickly obtain tentative identification of isolates as to species or species complexes. ?? 2009 The Society for Applied Microbiology.

  6. Effects of entomopathogenic fungi on different developmental stages of Cotesia flavipes (Cam. a parasitoid of Diatraea flavipennella (Box (Lepidoptera: Crambidae

    Directory of Open Access Journals (Sweden)

    Cinthia Conceição Matias da Silva

    2016-02-01

    Full Text Available Biological control using the parasitoid Cotesia flavipes (Cam. is one of the main components in the integrated management of the sugarcane moth borer Diatraea spp. Besides this parasitoid, the entomopathogenic fungi Beauveria bassiana (Bals. Vuill and Metarhizium anisopliae (Metsch. Sorok. are used to control sugarcane pests, and they can be found naturally parasitizing caterpillars. This study aimed to evaluate the effects of M. anisopliae and B. bassiana on different developmental stages of the C. flavipes parasitoid on Diatraea flavipennella (Box. The experiments were carried at laboratory with isolates PL 43 of M. anisopliae and ESALQ 447 of B. bassiana were applied to the parasitoid at the immature and adult phases. No negative effects were observed on the larval development of C. flavipes, and it could complete its development on caterpillars of D. flavipennella treated with fungi. The fungi did not cause pupal mortality. However, B. bassiana caused high mortality in the adult parasitoid (76%. The fungi had negative effects on parasitoids when applied during certain developmental stages of C. flavipes.

  7. Could insect phagocytic avoidance by entomogenous fungi have evolved via selection against soil amoeboid predators?

    Science.gov (United States)

    Bidochka, Michael J; Clark, David C; Lewis, Mike W; Keyhani, Nemat O

    2010-07-01

    The entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana are ubiquitously distributed in soils. As insect pathogens they adhere to the insect cuticle and penetrate through to the insect haemocoel using a variety of cuticle-hydrolysing enzymes. Once in the insect haemocoel they are able to survive and replicate within, and/or evade, phagocytic haemocyte cells circulating in the haemolymph. The mechanism by which these soil fungi acquire virulence factors for insect infection and insect immune avoidance is unknown. We hypothesize that insect phagocytic cell avoidance in M. anisopliae and B. bassiana is the consequence of a survival strategy against soil-inhabiting predatory amoebae. Microscopic examination, phagocytosis assays and amoeba mortality assays showed that these insect pathogenic fungi are phagocytosed by the soil amoeba Acanthamoeba castellanii and can survive and grow within the amoeba, resulting in amoeba death. Mammalian fungal and bacterial pathogens, such as Cryptococcus neoformans and Legionella pneumophila, respectively, show a remarkable overlap between survival against soil amoebae and survival against human macrophages. The insect immune system, particularly phagocytic haemocytes, is analogous to the mammalian macrophage. Our data suggest that the ability of the fungal insect pathogens M. anisopliae and B. bassiana to survive insect phagocytic haemocytes may be a consequence of adaptations that have evolved in order to avoid predation by soil amoebae.

  8. 培养基和温度对2株治蝗绿僵菌生长和毒力的影响%Effect of Different Media and Temperatures on Growth and Virulence of Two Isolates of Metarhizium anisopliaed from Locusta migr atoria manilensis

    Institute of Scientific and Technical Information of China (English)

    李树一; 雷仲仁; 李传仁

    2007-01-01

    研究了不同培养基及培养温度对治蝗绿僵菌(Metarhizium anisopliae)菌株LA和LD菌落面积、产孢量及毒力的影响,结果表明:LA在SMAY和PPDA培养基上的菌落面积较大,在SDAY上次之,在PCA上最小,且在SMAY培养基上的产孢量最高;LD在SMAY、PPDA和SDAY培养基上的菌落面积较大,在PCA上较小,且也在SMAY培养基上的产孢量最高;LA在30 ℃下生长较快,而LD则在25 ℃生长较快,但2种菌株均在25 ℃下产孢量较高;在SMAY上培养的LA、LD对东亚飞蝗(Locusta migratoria manilensis (Meyen))毒力较高,LT50分别是4.321 2 d和4.890 2 d.

  9. Metarhizium anisopliae CQMa102 biomass growth and extracellular phosphatase biosynthesis with different phosphorus sources%不同磷源对金龟子绿僵菌CQMa102生物量及生成胞外磷酸酶的影响

    Institute of Scientific and Technical Information of China (English)

    李鑫强; 朱培; 李振轮

    2013-01-01

    Metarhizium anisopliae is an important entomopathogenic fungus which is common and widely distributed in soil throughout the world.Some reports suggested that its extracellular phosphatase might play a pivotal role in invading and killing the hosts.This study explored the effect of different phosphorus sources on biomass growth and extracellular phosphatase biosynthesis of M.anisopliae by using shaking flask methods.Those evaluated phosphorus sources are including inorganic phosphorus (KH2PO4),simple organic phosphorus (phytate sodium and C6H5Na2PO4·2H2O) and protein phosphorus (casein).The results indicated that,in terms of biomass growth,production of acidic phosphatase and extracellular protein,casein yielded the best result,followed by KH2PO4 and C6H5Na2PO4·2H2O,wherein phytate sodium came at last.For purification of the extracellular protein phosphatase of M.anisopliae,the optimal phosphorus source is C6H5Na2PO4·2H2O,where there was the highest specific activity of protein phosphatase.%金龟子绿僵菌Metarhizium anisopliae是一种广泛分布于世界各地土壤中的重要的昆虫病原真菌.已有研究表明,胞外磷酸酶在绿僵菌侵染并致死寄主过程中发挥了重要作用.利用摇瓶培养方法探究了无机磷(KH2PO4)、简单有机磷(植酸钠、磷酸苯二钠)和蛋白有机磷(酪蛋白)为单独磷源条件下,绿僵菌生物量、产胞外酸性磷酸酶以及酪氨酸蛋白磷酸酶、丝/苏氨酸蛋白磷酸酶活性的变化.实验结果显示:加入酪蛋白的培养基,最有利于绿僵菌生长、胞外蛋白的分泌和产酸性磷酸酶;其次为加入KH2PO4和磷酸苯二钠的培养基;加入植酸钠的培养基不利于绿僵菌的生长代谢.然而,加入磷酸苯二钠的培养基,最有利于酪氨酸蛋白磷酸酶、丝/苏氨酸蛋白磷酸酶的分离纯化.

  10. Occurrence of entomopathogenic fungi from agricultural and natural ecosystems in Saltillo, México, and their virulence towards thrips and whiteflies.

    Science.gov (United States)

    Sánchez-Peña, Sergio R; Lara, Jorge San-Juan; Medina, Raúl F

    2011-01-01

    Entomopathogenic fungi were collected from soil in four adjacent habitats (oak forest, agricultural soil, pine reforestation and chaparral habitat) in Saltillo, México using the insect bait method with Tenebrio molitor (L.) (Coleoptera: Tenebrionidae) larvae as bait. Overall, of the larvae exposed to soil, 171 (20%) hosted Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Cordycipitaceae), 25 (3%) hosted Metarhizium anisopliae (Metschnikoff) Sorokin (Hypocreales: Clavicipitaceae) and 1 (0.1%) hosted lsaria (=Paecilomyces) sp. (Hypocreales: Cordycipitaceae). B. bassiana was significantly more frequent on larvae exposed to oak forest soil. M. anisopliae was significantly more frequent on larvae exposed to agricultural soil. From the infected bait insects, 93 isolates of B. bassiana and 24 isolates of M. anisopliae were obtained. Strains were tested for their infectivity against Cuban laurel thrips, Gynaikothrips uzeli Zimmerman (Thysanoptera: Phlaeothripidae) and the greenhouse whitefly, Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae). B. bassiana isolates caused the highest mortality on thrips (some causing 88% mortality after 6 days); both fungal species caused similarly high mortality levels against whiteflies (75%) after 6 days. Large amounts of germplasm of entomopathogenic fungi, fundamentally B. bassiana and M. anisopliae, exist in the habitats sampled; pathogenicity varied among strains, and some strains possessed significant virulence. Soils in these habitats are reservoirs of diverse strains with potential for use in biocontrol.

  11. A proposed role for the cuticular fatty amides of Liposcelis bostrychophila (Psocoptera: Liposcelidae) in preventing adhesion of entomopathogenic fungi with dry-conidia.

    Science.gov (United States)

    Lord, Jeffrey C; Howard, Ralph W

    2004-08-01

    Maximum challenge exposure of Liposcelis bostrychophila to Beauveria bassiana, Paecilomyces fumosoroseus, Aspergillus parasiticus or Metarhizium anisopliae resulted in no more than 16% mortality. We investigated several of L. bostrychophila's cuticular lipids for possible contributions to its tolerance for entomopathogenic fungi. Saturated C14 and C16 fatty acids did not reduce the germination rates of B. bassiana or M. anisopliae conidia. Saturated C6 to C12 fatty acids that have not been identified in L. bostrychophila cuticular extracts significantly reduced germination, but the reduction was mitigated by the presence of stearamide. Cis-6-hexadecenal did not affect germination rates. Mycelial growth of either fungal species did not occur in the presence of caprylic acid, was reduced by the presence of lauric acid, and was not significantly affected by palmitic acid. Liposcelis bostrychophila is the only insect for which fatty acid amides have been identified as cuticular components. Stearamide, its major fatty amide, did not reduce germination of B. bassiana or M. anisopliae conidia or growth of their mycelia. Adhesion of conidia to stearamide preparations did not differ significantly from adhesion to the cuticle of L. bostrychophila. Pretreatment of a beetle known to be fungus-susceptible, larval Oryzaephilus surinamensis, with stearamide significantly decreased adhesion of B. bassiana or M. anisopliae conidia to their cuticles. This evidence indicates that cuticular fatty amides may contribute to L. bostrychophila's tolerance for entomopathogenic fungi by decreasing hydrophobicity and static charge, thereby reducing conidial adhesion.

  12. Field Applications of Entomopathogenic Fungi Beauveria bassiana (Hypocreales: Clavicipitaceae) and Metarhizium anisopliae F52 (Hypocreales: Nectriaceae) for the Control of Ixodes scapularis (Acari: Ixodidae)

    Science.gov (United States)

    Two commercial formulations of Beauveria bassiana (Balsamo-Crivelli) Vuillemin were applied to residential sites in Old Lyme, Connecticut for the control of nymphs of the blacklegged tick, Ixodes scapularis in 1999 and 2000. The pyrethroid bifenthrin was applied to other homes for comparison with B....

  13. Debilitation in conidia of the entomopathogenic fungi Beauveria and Metarhizium anisopliae and implications with respect to viability determinations and mycopesticide quality assessments

    Science.gov (United States)

    Viabilities of entomopathogenic fungal conidia comprising biopesticide products are most commonly determined by suspension of dry conidia from storage in a water/surfactant solution immediately before inoculation onto an agar-based germination substrate; conidia are then incubated at a moderate temp...

  14. Use of the entomopathogenic fungi Metarhizium anisopliae, Cordyceps bassiana and Isaria fumosorosea to control Diaphorina citri (Hemiptera: Psylidae) in Persian lime under field conditions

    Science.gov (United States)

    The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is a destructive insect pest in the citriculture, because it is an efficient vector of the proteobacteria, ‘Candidatus Liberibacter asiaticus’ (Las), ‘Ca. L. Africanus’ (Laf), and ‘Ca. L. Americanus’ (Lam). These bacteria c...

  15. Selection of Entomopathogenic Fungi to Control Varroa destructor (Acari: Varroidae Selección de Hongos Entomopatógenos para el Control de Varroa destructor (Acari: Varroidae

    Directory of Open Access Journals (Sweden)

    Marta Rodríguez

    2009-12-01

    Full Text Available The aim of this work was select entomopathogenic fungi tolerant to temperatures inside the brood area of honey bees (Apis mellifera for to control Varroa destructor. For this purpose, 50 Beauveria bassiana (Balsamo Vuillemin and 48 Metarhizium anisopliae (Metschn. Sorokin isolates were evaluated at 30 and 35 ºC. For each isolate, colony discs of 5 mm with mycelium were placed in the center of a Petri dish with Sabouraud dextrose agar (SDA medium. The dishes were incubated at 30 and 35 °C, without light. Radial growth of each colony was measured daily. All the B. bassiana and M. anisopliae isolates presented a lineal growth rate at a temperature of 30 ºC. However, at 35 ºC, most of the isolates did not grow, except three B. bassiana and 14 M. anisopliae isolates (P El objetivo de este trabajo fue seleccionar hongos entomopatógenos tolerantes a las temperaturas del nido de cría de las abejas (Apis mellifera, para ser utilizados en el control de Varroa destructor. Se evaluaron 50 aislamientos de Beauveria bassiana (Balsamo Vuillemin y 48 de Metarhizium anisopliae (Metschn. Sorokin a temperaturas de 30 y 35 ºC. Discos de agar de 5 mm de diámetro con micelio de colonias de cada aislamiento, se depositaron en el centro de placas Petri con medio agar Sabouraud dextrosa (ASD. Las placas fueron incubadas a 30 y 35 °C y oscuridad y diariamente se midió el radio de cada colonia. Todos los aislamientos de B. bassiana y M. anisopliae var. anisopliae presentaron una tendencia lineal a través del tiempo a temperaturas de incubación de 30 °C. A 35 °C la mayoría de los aislamientos no crecieron, excepto tres aislamientos de B. bassiana y 14 de M. anisopliae (p < 0,001. Estos aislamientos fueron seleccionados para realizar pruebas de patogenicidad sobre V. destructor, aplicando una suspensión de 10(7 conidias mL-1. El aislamiento más efectivo fue Qu-M845 de M. anisopliae (p = 0,0033, produjo una mortalidad de 85%. La capacidad patogénica de este

  16. Metarhizium brunneum Blastospore Pathogenesis in Aedes aegypti Larvae: Attack on Several Fronts Accelerates Mortality.

    Science.gov (United States)

    Alkhaibari, Abeer M; Carolino, Aline T; Yavasoglu, Sare I; Maffeis, Thierry; Mattoso, Thalles C; Bull, James C; Samuels, Richard I; Butt, Tariq M

    2016-07-01

    Aedes aegypti is the vector of a wide range of diseases (e.g. yellow fever, dengue, Chikungunya and Zika) which impact on over half the world's population. Entomopathogenic fungi such as Metarhizium anisopliae and Beauveria bassiana have been found to be highly efficacious in killing mosquito larvae but only now are the underlying mechanisms for pathogenesis being elucidated. Recently it was shown that conidia of M. anisopliae caused stress induced mortality in Ae. aegypti larvae, a different mode of pathogenicity to that normally seen in terrestrial hosts. Blastospores constitute a different form of inoculum produced by this fungus when cultured in liquid media and although blastospores are generally considered to be more virulent than conidia no evidence has been presented to explain why. In our study, using a range of biochemical, molecular and microscopy methods, the infection process of Metarhizium brunneum (formerly M. anisopliae) ARSEF 4556 blastospores was investigated. It appears that the blastospores, unlike conidia, readily adhere to and penetrate mosquito larval cuticle. The blastospores are readily ingested by the larvae but unlike the conidia are able infect the insect through the gut and rapidly invade the haemocoel. The fact that pathogenicity related genes were upregulated in blastospores exposed to larvae prior to invasion, suggests the fungus was detecting host derived cues. Similarly, immune and defence genes were upregulated in the host prior to infection suggesting mosquitoes were also able to detect pathogen-derived cues. The hydrophilic blastospores produce copious mucilage, which probably facilitates adhesion to the host but do not appear to depend on production of Pr1, a cuticle degrading subtilisin protease, for penetration since protease inhibitors did not significantly alter blastospore virulence. The fact the blastospores have multiple routes of entry (cuticle and gut) may explain why this form of the inoculum killed Ae. aegypti larvae

  17. Metarhizium brunneum Blastospore Pathogenesis in Aedes aegypti Larvae: Attack on Several Fronts Accelerates Mortality

    Science.gov (United States)

    Alkhaibari, Abeer M.; Carolino, Aline T.; Yavasoglu, Sare I.; Maffeis, Thierry; Mattoso, Thalles C.; Bull, James C.; Samuels, Richard I.; Butt, Tariq M.

    2016-01-01

    Aedes aegypti is the vector of a wide range of diseases (e.g. yellow fever, dengue, Chikungunya and Zika) which impact on over half the world’s population. Entomopathogenic fungi such as Metarhizium anisopliae and Beauveria bassiana have been found to be highly efficacious in killing mosquito larvae but only now are the underlying mechanisms for pathogenesis being elucidated. Recently it was shown that conidia of M. anisopliae caused stress induced mortality in Ae. aegypti larvae, a different mode of pathogenicity to that normally seen in terrestrial hosts. Blastospores constitute a different form of inoculum produced by this fungus when cultured in liquid media and although blastospores are generally considered to be more virulent than conidia no evidence has been presented to explain why. In our study, using a range of biochemical, molecular and microscopy methods, the infection process of Metarhizium brunneum (formerly M. anisopliae) ARSEF 4556 blastospores was investigated. It appears that the blastospores, unlike conidia, readily adhere to and penetrate mosquito larval cuticle. The blastospores are readily ingested by the larvae but unlike the conidia are able infect the insect through the gut and rapidly invade the haemocoel. The fact that pathogenicity related genes were upregulated in blastospores exposed to larvae prior to invasion, suggests the fungus was detecting host derived cues. Similarly, immune and defence genes were upregulated in the host prior to infection suggesting mosquitoes were also able to detect pathogen-derived cues. The hydrophilic blastospores produce copious mucilage, which probably facilitates adhesion to the host but do not appear to depend on production of Pr1, a cuticle degrading subtilisin protease, for penetration since protease inhibitors did not significantly alter blastospore virulence. The fact the blastospores have multiple routes of entry (cuticle and gut) may explain why this form of the inoculum killed Ae. aegypti

  18. Metarhizium brunneum Blastospore Pathogenesis in Aedes aegypti Larvae: Attack on Several Fronts Accelerates Mortality.

    Directory of Open Access Journals (Sweden)

    Abeer M Alkhaibari

    2016-07-01

    Full Text Available Aedes aegypti is the vector of a wide range of diseases (e.g. yellow fever, dengue, Chikungunya and Zika which impact on over half the world's population. Entomopathogenic fungi such as Metarhizium anisopliae and Beauveria bassiana have been found to be highly efficacious in killing mosquito larvae but only now are the underlying mechanisms for pathogenesis being elucidated. Recently it was shown that conidia of M. anisopliae caused stress induced mortality in Ae. aegypti larvae, a different mode of pathogenicity to that normally seen in terrestrial hosts. Blastospores constitute a different form of inoculum produced by this fungus when cultured in liquid media and although blastospores are generally considered to be more virulent than conidia no evidence has been presented to explain why. In our study, using a range of biochemical, molecular and microscopy methods, the infection process of Metarhizium brunneum (formerly M. anisopliae ARSEF 4556 blastospores was investigated. It appears that the blastospores, unlike conidia, readily adhere to and penetrate mosquito larval cuticle. The blastospores are readily ingested by the larvae but unlike the conidia are able infect the insect through the gut and rapidly invade the haemocoel. The fact that pathogenicity related genes were upregulated in blastospores exposed to larvae prior to invasion, suggests the fungus was detecting host derived cues. Similarly, immune and defence genes were upregulated in the host prior to infection suggesting mosquitoes were also able to detect pathogen-derived cues. The hydrophilic blastospores produce copious mucilage, which probably facilitates adhesion to the host but do not appear to depend on production of Pr1, a cuticle degrading subtilisin protease, for penetration since protease inhibitors did not significantly alter blastospore virulence. The fact the blastospores have multiple routes of entry (cuticle and gut may explain why this form of the inoculum killed

  19. 田间噻嗪酮与黄绿绿僵菌对褐飞虱的协同防治%Synergism Between Low Concentration Buprofezin and Metarhizium anisopliae var. acridum on Controlling Nilaparvata lugens St(a)l(Homoptera: Delphacidae)

    Institute of Scientific and Technical Information of China (English)

    耿博闻; 张润杰

    2005-01-01

    在广州郊区稻田中用黄绿绿僵菌Metarhizium anisopliae var.acridum Bosenberg et Strand和低用量农药噻嗪酮(buprofezin)对褐飞虱Nilaparvata lugens Stl进行了协同防治作用药效试验.试验验共设低用量噻嗪酮、高用量噻嗪酮、黄绿绿僵菌、l低用量噻嗪酮和黄绿绿僵菌混施、清水对照共5个喷雾处理,每处理3个重复小区(4 m×5 m),随机区组排列.在处理前1天、处理后的第2、6、11、16、21 d分别调查褐飞虱每丛的飞虱数.结果表明,低用量噻嗪酮与黄绿绿僵菌混施可以有效的控制田间稻飞虱的种群密度,对成虫的防效在第16天、对若虫第21天时达到95%以上,其相对防效和虫口减退率都有明显升高并表现出明显的协同作用,这种协同作用对褐飞虱成虫的防治效果优于对褐飞虱若虫的防治效果.

  20. 绿僵菌侵染小菜蛾幼虫过程的透射电镜观察%Observation of infection process of Metarhizium anisopliae on Plutella xylostella larvae with transmission electron microscopy

    Institute of Scientific and Technical Information of China (English)

    王音; 雷仲仁; 张青文; 问锦曾

    2006-01-01

    利用透射电镜观察了绿僵菌Metarhizium anisopliae对小菜蛾Plutella xylostella幼虫的侵染过程.结果表明:接种后22 h最早观察到绿僵菌穿透到寄主表皮层中,在穿透表皮过程中伴随着机械力和酶的活动.菌体进入寄主血腔后,以菌丝出芽生殖、菌丝分隔及菌丝段分隔3种方式大量增殖,主要形成颗粒状的菌丝段在寄主体腔内扩散,也形成少量的丝状菌丝.

  1. Sequence analysis of the rDNA intergenic spacer of Metarhizium strains isolated in Brazil

    Directory of Open Access Journals (Sweden)

    Fabiana Y. Yanaka-Schäfer

    2008-01-01

    Full Text Available To assess the extent of genetic variability of rDNA intergenic spacer (IGS in Metarhizium sp., 34 strains (27 isolated in Brazil were sequenced and analyzed together with an additional 20 Metarhizium anisopliae var. anisopliae sequences retrieved from GenBank. Overall, the global nucleotide diversity for the region under study was of 0.090, while for the Brazilian isolates it was only 0.016. Phylogenetic analyses showed four well-supported groups (A, B, C, and D, one of which (D has not been previously identified. All but one of the Brazilian strains cluster in this novel D phylogroup, suggesting that the genetic variation found in Brazil is a subset of the worldwide M. anisopiliae var. anisopliae variation.

  2. Trials to control South American ticks with entomopathogenic fungi.

    Science.gov (United States)

    Bittencourt, V R

    2000-01-01

    Ticks are parasites of great importance to man, domestic, and wild animals. In spite of the progress of the science, thus far, ticks are controlled mainly by chemical acaricides and when this technique is used in an indiscriminate way it can lead to ambiental pollution and development of tick resistance to acarcides. The studies developed in the Universidade Federal Rural do Rio de Janeiro demonstrate that some isolates of the fungi Metarhizium anisopliae and Beauveria bassiana are pathogens for ticks and their action effects the mortality of ticks and reduces the populational levels in subsequent generations. In future, research needs to address the search of new pathogens; the association of biological with chemical products seeking an synergistic action; the search of formulations that propitiate increased stability, and careful studies on the safety of biological products for use in animals.

  3. Isolation and identification of Metarhizium from Citrus grandis ‘tomentosa’ GAP base%化橘红GAP基地土壤绿僵菌的分离鉴定

    Institute of Scientific and Technical Information of China (English)

    马维思; 徐江; 乔海莉; 陈君; 李向明; 覃蓉敏; 程惠珍

    2012-01-01

    A type of entomopathogenic fungus of soil in Citrus grandis 'tomentosa' production base was isolated and identified with morphological and molecular biological methods, including pathogenesis, spore characteristic and ITS sequence analysis were conducted. The results showed that eighteen entomopathogenic fungi strains were isolated from the Tenebrio molitor infected in the soil samples, which were identified as Metarhizium anhopliae var. anisopliae. Based on results above, we concluded that there was quantity of Metarhaium resources in this area. These provided the useful information for controlling some pests of C grandis by using these strains of fungus.%目的:从化橘红GAP基地土壤中分离、鉴定绿僵菌,为进一步利用绿僵菌防治化橘红害虫奠定基础.方法:采用黄粉虫诱集法从40份土壤样品中诱集、分离绿僵菌,通过形态特征和ITS序列比对鉴定其种类.结果:从化橘红GAP基地土壤中共分离出18个绿僵菌菌株,经鉴定均为金龟子绿僵菌小孢变种Metarhizium anisopliae var.anisoplia.结论:化橘红GAP基地环境适于金龟子绿僵菌小孢变种生存,土壤中拥有丰富的绿僵菌资源,这为进一步利用绿僵菌防治化橘红害虫奠定基础.

  4. Entomopathogenic fungi disturbed the larval growth and feeding performance of Ocinara varians (Lepidoptera: Bombycidae) larvae

    Institute of Scientific and Technical Information of China (English)

    Abid Hussain; Ming-Yi Tian; Yu-Rong He; Sohail Ahmed

    2009-01-01

    Feeding experiments using three strains of entomopathogenic fungi, Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea were conducted with newly moulted 3rd-5th instar Ocinara varians Walker larvae in the laboratory. The mortality of larvae immersed individually in spore suspension (1×10~7 spores/mL) of all the strains was > 80% except 5th instar larvae treated with M. anisopliae which transformed into pupae, but did not result in adult emergence. The growth (total body mass), consumption, relative consumption rate and relative growth rate, were reduced at all three larval stages, while developmental time was extended in infected larvae with concurrent significant increase in approximate digestibility in infected larvae. Conversion of digested food (ECD) and ingested food (ECI) values declined in infected larvae as compared to the healthy larvae (control). The 5th instar larvae treated with M. anisopliae showed higher ECD and ECI values than control. Based on mortality and growth inhibition it can be suggested that all the studied fungal strains have a high potential for biocontrol and could be developed into biocontrol agents against O. varians.

  5. Changes of some biochemical estimates in the hemolymph and body wall of Dendrolimus punctatus infected by Metarhizium anisopliae%马尾松毛虫感染绿僵菌后某些生化指标的变化

    Institute of Scientific and Technical Information of China (English)

    宋漳; 冯丽贞; 景云

    2002-01-01

    3~4龄马尾松毛虫Dendrolimus punctatus幼虫被金龟子绿僵菌Metarhizium anisopliae感染后1~4d,血淋巴中血细胞总数和可溶性蛋白浓度均显著高于同期未感染的幼虫.利用聚丙烯酰胺凝胶电泳技术,分析了感染后3,4,6 d幼虫体壁组织中可溶性蛋白和过氧化物酶,以及血淋巴中过氧化物酶的变化.结果显示,金龟子绿僵菌的侵染对马尾松毛虫幼虫体壁组织蛋白质代谢产生影响,过氧化物酶活性有减弱趋势.

  6. Immune response of Chilo suppressalis Walker (Lepidoptera: Crambidae) larvae to different entomopathogenic fungi.

    Science.gov (United States)

    Zibaee, A; Malagoli, D

    2014-04-01

    The current study reports mortality and effects on cellular immune response of several entomopathogenic fungi including isoleates BB1, BB2 and BB3 of Beauveria bassiana, Metarhizium anisopliae, Isaria fumosoroseus and Lecanicilium lecanii against larvae of Chilo suppressalis. Prohemocytes, granulocytes, plasmatocytes and oenocytoids were identified as the main circulating hemocytes in the hemolymph of larvae using Giemsa staining solution. Entomopathogenic fungi caused differential mortality on larvae: BB1, BB3, M. anisopliae lead to the highest mortality on larvae and L. lecanii caused the lowest mortality. The highest numbers of total hemocytes were observed 3 h post-injection of B. bassiana isolates and 6 h for the other treatments. The highest numbers of plasmatocytes were observed 3 h post-injection of BB1 and Tween 80, whereas BB2, BB3, M. anisopliae, I. fumosoroseus and L. lecani caused plasmatocyte increase 6 h post-injection. Similar results were obtained in case of granulocytes but only Tween 80 showed the highest number of hemocytes 3 h post-injection. The highest numbers of nodules were found at various time intervals after injection of fungal isolates and latex bead. The highest activities of phenoloxidase were observed 12 h post-injection by BbB1, BbB3, M. anisopliae and latex bead; 3-6 h post-injection by BbB2, 6 h post-injection by I. fumosoroseus and 3-6 h post-injection by L. lecanii. Our data demonstrate the possibility of utilizing different fungal extracts in the field to help reduce the risk of resistance evolution in C. suppressalis and encourage experimentations aimed to increase the number of biological control agent for insect pests such as the striped rice stem borer C. suppressalis.

  7. Pathogenicity, Ovicidal Action, and Median Lethal Concentrations (LC50 of Entomopathogenic Fungi against Exotic Spiralling Whitefly, Aleurodicus dispersus Russell

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    Boopathi Thangavel

    2013-01-01

    Full Text Available Biological control using entomopathogenic fungi could be a promising alternative to chemical control. Entomopathogenic fungi, Beauveria bassiana (Balsamo Vuillemin, Metarhizium anisopliae (Metschnikoff Sorokin, Lecanicillium lecanii (Zimmerm. Zare and Gams, and Paecilomyces fumosoroseus (Wize Brown and Smith, were tested for their pathogenicity, ovicidal effect, and median lethal concentrations (LC50 against exotic spiralling whitefly, Aleurodicus dispersus Russell. The applications were made at the rate of 2 × 109 conidia mL−1 for evaluating the pathogenicity and ovicidal effect of entomopathogenic fungi against A. dispersus. The results of pathogenicity test showed that P. fumosoroseus (P1 strain was highly pathogenic to A. dispersus recording 100% mortality at 15 days after treatment (DAT. M. anisopliae (M2 strain had more ovicidal effect causing 37.3% egg mortality at 8 DAT. However, L. lecanii (L1 strain caused minimum egg hatchability (23.2% at 10 DAT as compared to control (92.6%. The lowest LC50 produced by P. fumosoroseus (P1 strain as 8.189 × 107 conidia mL−1 indicated higher virulence against A. dispersus. Hence, there is potential for use of entomopathogenic fungi in the field conditions as an alternate control method in combating the insect pests and other arthropod pests since they are considered natural mortality agents and are environmentally safe.

  8. Pathogenicity, Ovicidal Action, and Median Lethal Concentrations (LC 50 ) of Entomopathogenic Fungi against Exotic Spiralling Whitefly, Aleurodicus dispersus Russell.

    Science.gov (United States)

    Thangavel, Boopathi; Palaniappan, Karuppuchamy; Manickavasagam Pillai, Kalyanasundaram; Subbarayalu, Mohankumar; Madhaiyan, Ravi

    2013-01-01

    Biological control using entomopathogenic fungi could be a promising alternative to chemical control. Entomopathogenic fungi, Beauveria bassiana (Balsamo) Vuillemin, Metarhizium anisopliae (Metschnikoff) Sorokin, Lecanicillium lecanii (Zimmerm.) Zare and Gams, and Paecilomyces fumosoroseus (Wize) Brown and Smith, were tested for their pathogenicity, ovicidal effect, and median lethal concentrations (LC50) against exotic spiralling whitefly, Aleurodicus dispersus Russell. The applications were made at the rate of 2 × 10(9) conidia mL(-1) for evaluating the pathogenicity and ovicidal effect of entomopathogenic fungi against A. dispersus. The results of pathogenicity test showed that P. fumosoroseus (P1 strain) was highly pathogenic to A. dispersus recording 100% mortality at 15 days after treatment (DAT). M. anisopliae (M2 strain) had more ovicidal effect causing 37.3% egg mortality at 8 DAT. However, L. lecanii (L1 strain) caused minimum egg hatchability (23.2%) at 10 DAT as compared to control (92.6%). The lowest LC50 produced by P. fumosoroseus (P1 strain) as 8.189 × 10(7) conidia mL(-1) indicated higher virulence against A. dispersus. Hence, there is potential for use of entomopathogenic fungi in the field conditions as an alternate control method in combating the insect pests and other arthropod pests since they are considered natural mortality agents and are environmentally safe.

  9. Effects of steam-distilled shoot extract of Tagetes minuta (Asterales: Asteraceae) and entomopathogenic fungi on larval Tetanops myopaeformis.

    Science.gov (United States)

    Dunkel, Florence V; Jaronski, Stefan T; Sedlak, Christopher W; Meiler, Svenja U; Veo, Kendra D

    2010-06-01

    Interactions of a biopesticidal formulation of steam distilled shoot extract of Mexican marigold, Tagetes minuta, and entomopathogenic fungi were evaluated for management of the sugarbeet root maggot, Tetanops myopaeformis (Röder). Shoot extract plus surfactant (E-Z Mulse) (=T. minuta oil) was used in a 65:35 ratio to test the hypothesis that this fungicidal and nematocidal biopesticide causes dose-dependent mortality and developmental arrest of T. myopaeformis but does not interfere with the action of entomopathogenic fungi when applied together. A soil-petri dish bioassay system was developed to test the hypothesis. For diapausing, nonfeeding but active 12-mo-old third-instar larvae, 0.5% T. minuta oil treatment (=0.325% active ingredient [AI]) was sufficient to prevent pupation without mortality, but 0.75% T. minuta oil treatment (=0.458% AI) was lethal for 93% of the test insects. The effect of T. minuta oil on fungal efficacy under simultaneous use was studied using a model system of two entomopathogenic fungi, Beauveria bassiana (Bals.) Vuillemin. TM28 and Metarhizium anisopliae variety anisopliae (Metsch.) Sorokin MA 1200, in a soil-based bioassay with larval sugarbeet root maggots. No adverse effects of T. minuta oil on action of entomopathogenic fungi and no synergy were found; an additive effect of the T. minuta oil and each fungal isolate separately was found.

  10. Factors affecting the occurrence and distribution of entomopathogenic fungi in natural and cultivated soils.

    Science.gov (United States)

    Quesada-Moraga, Enrique; Navas-Cortés, Juan A; Maranhao, Elizabeth A A; Ortiz-Urquiza, Almudena; Santiago-Alvarez, Cándido

    2007-08-01

    Factors affecting the occurrence and distribution of entomopathogenic fungi in 244 soil samples collected from natural and cultivated areas in Spain were studied using an integrated approach based on univariate and multivariate analyses. Entomopathogenic fungi were isolated from 175 of the 244 (71.7%) soil samples, with only two species found, Beauveria bassiana and Metarhizium anisopliae. Of the 244 soil samples, 104 yielded B. bassiana (42.6%), 18 yielded M. anisopliae (7.3%), and 53 soil samples (21.7%) harboured both fungi. Log-linear models indicated no significant effect of habitat on the occurrence of B. bassiana, but a strong association between M. anisopliae and soils from cultivated habitats, particularly field crops. Also, irrespective of habitat type, B. bassiana predominated over M. anisopliae in soils with a higher clay content, higher pH, and lower organic matter content. Logistic regression analyses showed that pH and clay content were predictive variables for the occurrence of B. bassiana, whereas organic matter content was the predictive variable for M. anisopliae. Also, latitude and longitude predicted the occurrence of these same species, but in opposite directions. Altitude was found to be predictive for the occurrence of B. bassiana. Using principal component analysis, four factors (1 to 4) accounted for 86% of the total variance; 32.8, 22.9, 19.6 and 10.4% of the cumulative variance explained, respectively. Factor 1 was associated with high positive weights for soil clay and silt content and high negative weights for soil sand content. Factor 2 was associated with high positive weights for soil organic matter content and high negative weights for soil pH. Factor 3 was associated with high positive weights for latitude and longitude of the sampled localities and factor 4, had high positive weights only for the altitude. Bi-plot displays representing soil samples were developed for different factor combinations and indicated that, irrespective

  11. Physiological changes in Rhipicephalus microplus (Acari: Ixodidae) experimentally infected with entomopathogenic fungi.

    Science.gov (United States)

    Angelo, Isabele C; Tunholi-Alves, Vinícius M; Tunholi, Victor M; Perinotto, Wendell M S; Gôlo, Patrícia S; Camargo, Mariana G; Quinelato, Simone; Pinheiro, Jairo; Bittencourt, Vânia R E P

    2015-01-01

    Carbohydrate metabolism plays an important role in the physiology and maintenance of energy stores within living organisms. However, when organisms are exposed to adverse physiological conditions, such as during pathogenic infection, these organisms begin to use alternative substrates (proteins and lipids) for energy production. This paper studied the carbohydrate metabolism of Rhipicephalus microplus after infection with Beauveria bassiana and Metarhizium anisopliae. The parameters evaluated were glucose concentration, enzymatic activities of lactate dehydrogenase (LDH), alanine aminostransferase (ALT) and aspartate aminostransferase (AST), amounts of uric acid and urea in the hemolymph, and amount of glycogen in the fat body. The results showed changes in nitrogenous products, including an increase in the amount of urea detected 48 h after infection with both fungi. The enzymatic activities of LDH, ALT, and AST were increased after infection. The amount of glucose was increased 24 h after infection with B. bassiana and was reduced 48 h after infection with both fungi. The amount of glycogen in the fat body was reduced at different times of infection with both fungi. These results demonstrate, for the first time, the changes in carbohydrate metabolism of R. microplus after infection with M. anisopliae and B. bassiana and contribute to a better understanding of this host-parasite relationship. Together with knowledge of diseases that affect these ticks and their susceptibility to entomopathogens, an understanding of tick physiology will be necessary for the effective implementation of current biological control methods and will assist in the discovery of new methods to control this ectoparasite.

  12. Selection of entomopathogenic fungi for aphid control.

    Science.gov (United States)

    Vu, Van Hanh; Hong, Suk Il; Kim, Keun

    2007-12-01

    Twelve strains of entomopathogenic fungi such as Lecanicillium lecanii, Paecilomyces farinosus, Beauveria bassiana, Metarhizium anisopliae, Cordyceps scarabaeicola, and Nomuraea rileyi were screened for aphid control. At 25 degrees C and 75% relative humidity (RH), among tested entomopathogenic fungi, L. lecanii 41185 showed the highest virulent pathogenicity for both Myzus persicae and Aphis gossypii, and their control values were both nearly 100% 5 and 2 d after treatment, respectively. Moreover, at an RH of 45% and in a wide temperature range (20-30 degrees C), L. lecanii 41185 also exhibited the highest virulence to M. persicae. The control value of M. persicae and the 50% lethal time (LT50) decreased significantly as the applied conidial concentration increased. The 50% lethal concentration (LC50) of the conidial suspension of this fungus was determined to be 6.55x10(5) conidia/ml. The control values of M. persicae resulting from the application of 1x10(7) and 1x10(8) conidia/ml were nearly the same and were significantly higher than that of 1x10(6) conidia/ml. The tested entomopathogenic fungi grew in a broad temperature range (15-30 degrees C). Lecanicillium strains showed optimum growth at 25 degrees C. The aerial conidia of Lecanicillium strains also could germinate in a broad temperature range (15-30 degrees C) and L. lecanii 41185 was the only strain with conidial germination at 35 degrees C.

  13. Pathogenicity of the Metarhizium anisopliae to Asias halodendri larvae at different temperature and humidity conditions%不同温湿度下绿僵菌对红缘天牛幼虫致病力的影响

    Institute of Scientific and Technical Information of China (English)

    汪敏捷; 刘强

    2014-01-01

    在实验室内测定不同温、湿度下绿僵菌Metarhizium anisopliae对红缘天牛Asias halodendri幼虫的致病力的影响.分别在5个温度、5个湿度梯度下,研究温湿度对绿僵菌对红缘天牛幼虫的致病力影响.28℃、相对湿度>90%是绿僵菌感染红缘天牛的最佳温度和湿度条件.在28℃条件下,5d后的幼虫的校正死亡率高达94.4%,其致死中时(LT50)约为8h,明显短于其他各组,且幼虫僵虫率最高.温度高于或低于28℃条件时,绿僵菌对红缘天牛幼虫致死率都会减弱,致死中时(LT50)均在33 h以上,僵虫率也降低.相对湿度越大,对幼虫致病力越强.相对湿度>90%时的致死效果最好,校正死亡率和僵虫率分别为99.3%和50%,此湿度条件下致死速率也最快,LT50仅为35.3 h;相对湿度50%-60%时,幼虫校正死亡率和僵虫率显著降低,LT50约为122 h.

  14. Prospects for the use of entomopathogenic fungi for control of stored-product pests%利用昆虫病原真菌防治贮粮害虫展望

    Institute of Scientific and Technical Information of China (English)

    杰夫· 劳尔德

    2007-01-01

    Only two fungus species,Beauveria bassiana and Metarhizium anisopliae,have been given serious attention as possible microbial controls for stored-product pests.Their host ranges are broad, but their potency for the various target insects varies greatly,and some of the most important pests such as the red flour beetle are very tolerant of fungi.Accordingly, strategies are needed to improve the fungal performance.One such strategy is combination with other environmentally benign treatments such as controlled desiccant dusts and controlled atmospheres. Ironically the relatively dryness of stored-product environments favors fungal efficacy.Desiccation stress renders some insects more vulnerable to fungi.The longevity of fungus spores is also best under dry conditions.The prospects of fungi for control of stored-product pests can be improved by taking advantage of these phenomena and judicious selection of use venues and application strategies.

  15. Production of microsclerotia by Brazilian strains of Metarhizium spp. using submerged liquid culture fermentation.

    Science.gov (United States)

    Mascarin, Gabriel Moura; Kobori, Nilce Naomi; de Jesus Vital, Rayan Carlos; Jackson, Mark Alan; Quintela, Eliane Dias

    2014-05-01

    We investigated the potential production and desiccation tolerance of microsclerotia (MS) by Brazilian strains of Metarhizium anisopliae (Ma), M. acridum (Mc) and M. robertsii (Mr). These fungi were grown in a liquid medium containing 16 g carbon l⁻¹ with a carbon:nitrogen ratio of 50:1. One hundred milliliters cultures were grown in 250 ml Erlenmeyer flasks in a rotary incubator shaker at 28 °C and 200 rpm for 5 days. Five-day-old MS were harvested, mixed with diatomaceous earth (DE) and air-dried for 2 days at 30 °C. The air-dried MS-DE granular preparations were milled by mortar + pestle and stored in centrifuged tubes at either 26 or -20 °C. Desiccation tolerance and conidia production were assessed for dried MS granules by measuring hyphal germination after incubation for 2 days on water agar plates at 26 °C and for conidia production following 7 days incubation. Yields of MS by all strains of Metarhizium were 6.1-7.3 × 10⁶ l⁻¹ after 3 days growth with maximum MS yields (0.7-1.1 × 10⁷ l⁻¹) after 5 days growth. No differences in biomass accumulation were observed after 3 days growth, whereas Ma-CG168 showed the highest biomass accumulation after 5 days growth. Dried MS-DE preparations of all fungal strains were equally tolerant to desiccation (≥93 % germination) and the highest conidia production was obtained by MS granules of Mc-CG423 (4 × 10⁹ conidia g⁻¹). All MS granules showed similar stability after storage at either 26 or -20 °C for 3.5 months.

  16. Occurrence of Metarhizium spp in central Brazilian soils

    Science.gov (United States)

    The biodiversity of entomopathogenic fungi in tropical ecosystems is still little investigated, and the objective of this study was to isolate and identify fungi of the entomopathogenic genus Metarhizium (Hypocreales: Clavicipitaceae) present in undisturbed soils of Central Brazilian Cerrado. A tota...

  17. Can insects develop resistance to insect pathogenic fungi?

    Directory of Open Access Journals (Sweden)

    Ivan M Dubovskiy

    Full Text Available Microevolutionary adaptations and mechanisms of fungal pathogen resistance were explored in a melanic population of the Greater wax moth, Galleria mellonella. Under constant selective pressure from the insect pathogenic fungus Beauveria bassiana, 25(th generation larvae exhibited significantly enhanced resistance, which was specific to this pathogen and not to another insect pathogenic fungus, Metarhizium anisopliae. Defense and stress management strategies of selected (resistant and non-selected (susceptible insect lines were compared to uncover mechanisms underpinning resistance, and the possible cost of those survival strategies. We hypothesize that the insects developed a transgenerationally primed resistance to the fungus B. bassiana, a costly trait that was achieved not by compromising life-history traits but rather by prioritizing and re-allocating pathogen-species-specific augmentations to integumental front-line defenses that are most likely to be encountered by invading fungi. Specifically during B. bassiana infection, systemic immune defenses are suppressed in favour of a more limited but targeted repertoire of enhanced responses in the cuticle and epidermis of the integument (e.g. expression of the fungal enzyme inhibitor IMPI, and cuticular phenoloxidase activity. A range of putative stress-management factors (e.g. antioxidants is also activated during the specific response of selected insects to B. bassiana but not M. anisopliae. This too occurs primarily in the integument, and probably contributes to antifungal defense and/or helps ameliorate the damage inflicted by the fungus or the host's own immune responses.

  18. Can insects develop resistance to insect pathogenic fungi?

    Science.gov (United States)

    Dubovskiy, Ivan M; Whitten, Miranda M A; Yaroslavtseva, Olga N; Greig, Carolyn; Kryukov, Vadim Y; Grizanova, Ekaterina V; Mukherjee, Krishnendu; Vilcinskas, Andreas; Glupov, Viktor V; Butt, Tariq M

    2013-01-01

    Microevolutionary adaptations and mechanisms of fungal pathogen resistance were explored in a melanic population of the Greater wax moth, Galleria mellonella. Under constant selective pressure from the insect pathogenic fungus Beauveria bassiana, 25(th) generation larvae exhibited significantly enhanced resistance, which was specific to this pathogen and not to another insect pathogenic fungus, Metarhizium anisopliae. Defense and stress management strategies of selected (resistant) and non-selected (susceptible) insect lines were compared to uncover mechanisms underpinning resistance, and the possible cost of those survival strategies. We hypothesize that the insects developed a transgenerationally primed resistance to the fungus B. bassiana, a costly trait that was achieved not by compromising life-history traits but rather by prioritizing and re-allocating pathogen-species-specific augmentations to integumental front-line defenses that are most likely to be encountered by invading fungi. Specifically during B. bassiana infection, systemic immune defenses are suppressed in favour of a more limited but targeted repertoire of enhanced responses in the cuticle and epidermis of the integument (e.g. expression of the fungal enzyme inhibitor IMPI, and cuticular phenoloxidase activity). A range of putative stress-management factors (e.g. antioxidants) is also activated during the specific response of selected insects to B. bassiana but not M. anisopliae. This too occurs primarily in the integument, and probably contributes to antifungal defense and/or helps ameliorate the damage inflicted by the fungus or the host's own immune responses.

  19. Tools for delivering entomopathogenic fungi to malaria mosquitoes: effects of delivery surfaces on fungal efficacy and persistence

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    Mnyone Ladslaus L

    2010-08-01

    Full Text Available Abstract Background Entomopathogenic fungi infection on malaria vectors increases daily mortality rates and thus represents a control measure that could be used in integrated programmes alongside insecticide-treated bed nets (ITNs and indoor residual spraying (IRS. Before entomopathogenic fungi can be integrated into control programmes, an effective delivery system must be developed. Methods The efficacy of Metarhizium anisopliae ICIPE-30 and Beauveria bassiana I93-825 (IMI 391510 (2 × 1010 conidia m-2 applied on mud panels (simulating walls of traditional Tanzanian houses, black cotton cloth and polyester netting was evaluated against adult Anopheles gambiae sensu stricto. Mosquitoes were exposed to the treated surfaces 2, 14 and 28 d after conidia were applied. Survival of mosquitoes was monitored daily. Results All fungal treatments caused a significantly increased mortality in the exposed mosquitoes, descending with time since fungal application. Mosquitoes exposed to M. anisopliae conidia on mud panels had a greater daily risk of dying compared to those exposed to conidia on either netting or cotton cloth (p B. bassiana conidia on mud panels or cotton cloth had similar daily risk of death (p = 0.14, and a higher risk than those exposed to treated polyester netting (p Conclusion Both fungal isolates reduced mosquito survival on immediate exposure and up to 28 d after application. Conidia were more effective when applied on mud panels and cotton cloth compared with polyester netting. Cotton cloth and mud, therefore, represent potential substrates for delivering fungi to mosquitoes in the field.

  20. Susceptibility of biological stages of the horn fly, Haematobia irritans, to entomopathogenic fungi (Hyphomycetes)

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    Angel-Sahagún, C. A.; Lezama-Gutiérrez, R.; Molina-Ochoa, J.; Galindo-Velasco, E.; López-Edwards, M.; Rebolledo-Domínguez, O.; Cruz-Vázquez, C.; Reyes-Velázquez, W. P.; Skoda, S. R.; Foster, J. E.

    2005-01-01

    The susceptibility of the egg, pupa, and adult of Haematobia irritans (L.) (Diptera: Muscidae) to isolates of the fungi Metarhizium anisopliae (Metsch.) Sor., Beauveria bassiana (Bals.) Vuill., and Paecilomyces fumosoroseus (Wize) Brown and Smith, was evaluated under laboratory conditions. Groups of 20 eggs than 4 h old, pupae less than 48h old and adults were sprayed with a conidial suspension of each isolate. Eggs, pupae and adults of horn fly were susceptible to these entomopathogenic fungi. For treated eggs, the isolates Ma3, Ma 15, Ma25, Pfr1, and Pfr8 reduced adult emergence to 3.8% to 6.3% in comparison with the control (72%). The mortality of pupae infected by the isolates Ma2, Ma25, and Pfr10 ranged between 50% and 71.3%. Mortality of adults after treatment with the isolates Ma6, Ma 10, Ma 14, Ma 15, Pfr 1, Pfr 9, Pfr 10, Pfr 11, and Pfr12 were higher than 90%. The isolate Ma6 produced the lowest LC50 against adult horn flies (8.08 × 102conidia/ml). These findings supported the hypotheses that isolates of M. anisopliae, and P. fumosoroseus are pathogenic against the different biological stages of horn flies by reducing adult emergence when applied on groups of eggs and pupae, and producing mortality when applied to adults. PMID:17119632

  1. Susceptibility of different populations of ticks to entomopathogenic fungi.

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    Perinotto, W M S; Angelo, I C; Golo, P S; Quinelato, S; Camargo, M G; Sá, F A; Bittencourt, V R E P

    2012-03-01

    This study aimed to evaluate the in vitro effect of the entomopathogenic fungi Metarhizium anisopliae sensu lato (s.l.) and Beauveria bassiana sensu lato (s.l.) on two distinct populations of Rhipicephalus microplus, from two different experimental farms. Bioassays were performed with engorged females, eggs and larvae. Fungal infection was evaluated based on biological parameters of treated engorged females, percentage of hatch from treated eggs, and percentage of mortality and mean lethal time (LT90) of treated larvae. When the treatments were compared between the two populations, there were significant differences in the following parameters: pre-oviposition period, hatching period and egg production index. Moreover, the results showed that B. bassiana s.l., isolate Bb 986, was more virulent than M. anisopliae s.l., isolate 959, for engorged females, showing a control percentage of 49%. In the bioassay with eggs, the hatching percentage ranged from 3.1% to 49.5% in one population and from 3.4% to 42.7% in the other, with no significant difference between the two populations. In the bioassay with unfed larvae, the mortality percentage ranged from 91.8% to 98.7% in one population and from 71.0% to 94.0% in the other. The LT(90) varied from 19.52 to 27.51 days in one of the populations and 22.89 to 37.31 days in the other. These results suggest that populations of R. microplus show distinct variation in their susceptibility to B. bassiana s.l. and M. anisopliae s.l.

  2. Molecular, morphological and pathogenic characterization of six strains of Metarhizium spp. (Deuteromycotina: Hyphomycetes for the control of Aegorhinus superciliosus (Coleoptera: Curculionidae

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    María Sepúlveda

    2016-03-01

    Full Text Available Aegorhinus superciliosus is an important pest on blueberry (Vaccinium corymbosum L. and other fruit trees. The use of entomopathogenic fungi as Metarhizium spp. has been evaluated for the control of this insect, but variability has been observed among different strains. The aim of this study was to characterize six promising strains of Metarhizium spp. for the control of A. superciliosus. The studied strains were QuM173c, Qu-M363, Qu-M171a, Qu-M156a, Qu-M421, and Qu-M430, all of which belonged to the Chilean Collection of Microbial Genetic Resources (ChCMGR of the Institute de Investigaciones Agropecuarias (INIA, Chile. Molecular characterization was made by sequencing the ITS region (Internal Transcribed Spacers, ITS-5.8S rDNA. The morphology of conidia was evaluated through scanning electron microscopy and radial colony growth was evaluated in potato dextrose agar (PDA, Sabouraud dextrose agar (SDA, agar enriched with larvae of Galleria mellonella (Lepidoptera: Pyralidae (GA, and agar enriched with adults of A. superciliosus (AA. Pathogenicity was studied based on mortality of adults of A. superciliosus inoculated with conidia. Sequencing of the ITS-5.8S rDNA region indicates that the strains belong to the clade of M. anisopliae var. anisopliae, except for Qu-M171a, which was identified as M. anisopliae var. lepidiotum. Conidia average length for the six strains was 5.09 pm and average conidia width was 1.92 pm. Radial colony growth differences were observed between strains (p < 0.01 and between different growth media (p < 0.01. The strains exhibited the highest colony growth in the GA medium, while in the AA medium they showed the lowest (p < 0.01. Pathogenicity tests show that Qu-M430 reached a 90% mortality rate (p < 0.01. Results show that there is variability between the studied strains, which is expressed in their morphology, molecular characteristics and pathogenicity towards A. superciliosus.

  3. Virulence of Mexican isolates of entomopathogenic fungi (Hypocreales: Clavicipitaceae) upon Rhipicephalus=Boophilus microplus (Acari: Ixodidae) larvae and the efficacy of conidia formulations to reduce larval tick density under field conditions.

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    Angel-Sahagún, C A; Lezama-Gutiérrez, R; Molina-Ochoa, J; Pescador-Rubio, A; Skoda, S R; Cruz-Vázquez, C; Lorenzoni, A G; Galindo-Velasco, E; Fragoso-Sánchez, H; Foster, J E

    2010-06-24

    The first objective was laboratory evaluation of the virulence of 53 Mexican isolates of fungi against larvae of Rhipicephalus (Boophilus) microplus. Thirty-three isolates of Metarhizium anisopliae var. anisopliae (Metschnickoff) Sorokin (Hypocreales: Clavicipitaceae) and 20 isolates of Isaria (Paecilomyces) fumosorosea (fumosoroseus) (Wize) (Eurotiales: Trichomaceae) were tested on 7-day-old larvae under laboratory conditions. Larvae were immersed in a suspension containing 10(8)conidia/mL and the CL(50) values were estimated. Then, field tests were conducted to determine the efficacy of formulations of the isolate with the highest virulence. M. anisopliae (Ma 14 isolate) was formulated with four carriers: Tween, Celite, wheat bran, and Citroline (mineral oil) and applied on pasture beds of Cynodon plectostachyus (L.), at a dose of 2 x 10(9)CFU/m(2). In the first trial, M. anisopliae was applied on plots naturally infested with larvae; in the second trial, tick populations in the experimental plots were eliminated and then re-infested with 20,000 7-day-old larvae. In the laboratory, all M. anisopliae isolates infected larvae with a mortality range between 2 and 100%; also, 13 of 20 I. fumosorosea isolates caused mortality rates between 7 and 94%. In the first field trial, 14 days post-application, conidial formulations in Celite and wheat bran caused 67.8 and 94.2% population reduction, respectively. In the second trial, the Tween formulation caused the highest larval reduction, reaching up to 61% (28 days post-application). Wheat bran formulation caused 58.3% larval reduction (21 days post-application) and was one of the most effective. The carriers and emulsifiers have a large impact on the effectiveness of conidial formulations.

  4. Specific Diversity of Metarhizium Isolates Infecting Aeneolamia spp. (Hemiptera: Cercopidae) in Sugarcane Plantations.

    Science.gov (United States)

    Hernández-Domínguez, C; Guzmán-Franco, A W; Carrillo-Benítez, M G; Alatorre-Rosas, R; Rodríguez-Leyva, E; Villanueva-Jiménez, J A

    2016-02-01

    Spittlebugs from the genus Aeneolamia are important pests of sugarcane. Although the use of the entomopathogenic fungus Metarhizum anisopliae s.l. for control of this pest is becoming more common in Mexico, fundamental information regarding M. anisopliae in sugarcane plantations is practically non-existent. Using phylogenetic analysis, we determined the specific diversity of Metarhizium spp. infecting adult spittlebugs in sugarcane plantations from four Mexican states. We obtained 29 isolates of M. anisopliae s.str. Haplotype network analysis revealed the existence of eight haplotypes. Eight selected isolates, representing the four Mexican states, were grown at different temperatures in vitro; isolates from Oaxaca achieved the greatest growth followed by isolates from Veracruz, San Luis Potosi and Tabasco. No relationship was found between in vitro growth and haplotype diversity. Our results represent a significant contribution to the better understanding of the ecology of Metarhizum spp. in the sugarcane agroecosystem.

  5. Effect of oil-based formulations of acaripathogenic fungi to control Rhipicephalus microplus ticks under laboratory conditions.

    Science.gov (United States)

    Camargo, Mariana G; Golo, Patrícia S; Angelo, Isabele C; Perinotto, Wendell M S; Sá, Fillipe A; Quinelato, Simone; Bittencourt, Vânia R E P

    2012-08-13

    The formulations of acaripathogenic fungi to control ticks have been widely studied. The present study evaluated the efficacy of oil-based formulations of Metarhizium anisopliae sensu lato (s.l.), isolate Ma 959, and Beauveria bassiana, isolate Bb 986, on different Rhipicephalus microplus stages, comparing the efficacy between aqueous suspensions and 10, 15 and 20% mineral oil formulations. Twelve groups were formed: one aqueous control group; three mineral oil control groups, at 10, 15 or 20%; two aqueous fungal suspensions of M. anisopliae s.l. or B. bassiana; and three formulations of M. anisopliae (s.l.) or B. bassiana containing 10, 15, and 20% mineral oil. To prepare aqueous suspensions and oily formulations, fungal isolates were cultivated on rice grains in polypropylene bags. The conidial suspensions and formulations had a concentration of 10(8)conidia/mL. Bioassays were repeated twice. After treatment, the following biological parameters of engorged females were evaluated: hatching percentage, egg production index, nutritional index, and percentage of tick control. The following parameters were evaluated in the bioassays with eggs: period of incubation, period of hatch, and hatching percentage. Mortality was evaluated in bioassays with larvae. M. anisopliae s.l. and B. bassiana oil-based formulations were more effective than aqueous suspensions against R. microplus eggs, larvae and engorged females, however, there was no significant difference between the three oil concentrations used. M. anisopliae s.l. and B. bassiana formulated in mineral oil reached 93.69% and 21.67% efficacy, respectively, while M. anisopliae s.l. and B. bassiana aqueous suspensions attained 18.70% and 1.72% efficacy, respectively. M. anisopliae s.l. oil-based formulations caused significant effects in all biological parameters of engorged females while B. bassiana oil-based formulations modified significantly the nutritional index only. Eggs treated with M. anisopliae s.l. and B

  6. Interactions of two insect pathogens, Paranosema locustae (Protista: Microsporidia) and Metarhizium acridum (Fungi: Hypocreales), during a mixed infection of Locusta migratoria (Insecta: Orthoptera) nymphs.

    Science.gov (United States)

    Tokarev, Yuri S; Levchenko, Maxim V; Naumov, Anton M; Senderskiy, Igor V; Lednev, Georgiy R

    2011-02-01

    Locusta migratoria nymphs were fed Paranosema locustae spores and/or surface-treated with Metarhizium acridum 3 (assay 1), 6 (assay 2) or 9 days (assay 3) post microsporidia application (p.m.a.). These three dates corresponded to the key phases of P. locustae development: (a) mass proliferation, (b) transition to sporogenesis and (c) onset of spore maturation, respectively. As a result, locust mortality due to mixed treatment increased slower, equally and faster, as compared to mortality expected from the combination of two pathogens in assays 1-3, respectively. However, a statistically significant difference in survival times was observed only in assay 3, indicating that only at the phase of spore maturation microsporidia drastically increase locust susceptibility to fungal infection. Analysis of perished nymphs showed that fungal treatment 3 days p.m.a. impeded development of microsporidia. Fungal sporulation on locust cadavers was not affected by co-occurring microsporidiosis. Copyright © 2010 Elsevier Inc. All rights reserved.

  7. Entomopathogenic fungi as potential biocontrol agents of the ecto-parasitic mite, Varroa destructor, and their effect on the immune response of honey bees (Apis mellifera L.).

    Science.gov (United States)

    Hamiduzzaman, Mollah Md; Sinia, Alice; Guzman-Novoa, Ernesto; Goodwin, Paul H

    2012-11-01

    Three isolates of each of the entomopathogenic fungi, Metarhizium anisopliae, Beauveria bassiana and Clonostachys rosea, were assessed for their pathogenicity to the honey bee parasitic mite, Varroa destructor. The fungi were applied to varroa mites by immersing them in a spore solution, and then the inoculated mites were placed on honey bee brood inside capped cells. At 7 days post inoculation (dpi), the three fungi caused significant varroa mortality compared to non-inoculated mites. In brood treated only with varroa mites, expression of the honey bee genes, hymenoptaecin and poly U binding factor 68 Kd (pUf68), decreased over time, while expression of blue cheese (BlCh) and single minded (SiMd) was not affected. In brood inoculated directly only with M. anisopliae or B. bassiana, the emerged adults showed reduced weight indicating infection by the fungi, which was confirmed by observation of hyphae in the brood. Fungal infection of the brood resulted in increased expression of hymenoptaecin, pUf68 and BlCh, but not SiMd. In brood treated with varroa mites that had been inoculated with the fungi, expression of hymenoptaecin, pUf68 and BlCh, but not SiMd, was even more up-regulated. While varroa mites can suppress gene expression in honey bee brood, varroa mites infected with entomopathogenic fungi induced their expression. This may be due to a low level of fungal infection of the bee, which negated the immunosuppression by the mites. Therefore, entomopathogenic fungi could reduce varroa mite damage to honey bee brood by both infecting the parasite and preventing varroa-associated suppression of honey bee immunity.

  8. Laboratory evaluation of entomopathogenic fungi against larvae and adults of onion maggot (Diptera: Anthomyiidae).

    Science.gov (United States)

    Davidson, G; Chandler, D

    2005-12-01

    Laboratory experiments were done to measure the susceptibility of larvae and adults of the onion maggot, Delia antiqua (Meigen) (Diptera: Muscidae: Anthomyiidae) to 27 isolates of entomopathogenic fungi from four genera [Beauveria Vuillemin, Lecanicillium (Petch) Zare & W. Gams, Metarhizium Sorokin, and Paecilomyces Bainier]. A novel bioassay was developed for D. antiqua larvae by using a diet based on mixed vegetable powder. When evaluated in a virulence screen, the fungal isolates caused less mortality of D. antiqua larvae than adults. Only three isolates caused > 50% mortality of larvae, whereas 12 isolates caused > 50% mortality of adults. Fungal species was a statistically significant factor affecting the mortality of larvae but not of adults. The fungal isolates causing the most mortality of larvae tended to belong to Metarhizium anisopliae (Metschnikoff) Sorokin. Two M. anisopliae isolates (389.93 and 392.93) were evaluated in dose-response bioassays. The median lethal concentrations of the isolates against larvae were 6.1 x 10(7) conidia ml(-1) for isolate 389.93 and 7.6 x 10(7) conidia ml(-1) for isolate 392.93. The emergence of adult flies from pupae was reduced at high concentrations of conidia (3.0 x 10(8) and 1.0 x 10(8) conidia ml(-1)). The median lethal concentrations of the isolates against adults were 1.7 x 10(7) and 4.0 x 10(7) conidia ml(-1), respectively. Some of the fungal isolates examined may have potential as biological control agents of larvae of D. antiqua and related species.

  9. Evaluation of insect associated and plant growth promoting fungi in the control of cabbage root flies.

    Science.gov (United States)

    Razinger, Jaka; Lutz, Matthias; Schroers, Hans-Josef; Urek, Gregor; Grunder, Jürg

    2014-08-01

    Delia radicum L. or cabbage maggot is an important pest for Brassicaceous crops. There are currently no registered chemical control agents for its control in Slovenia. Fungal control agents for cabbage maggot were therefore sought among nine rhizosphere-compatible and plant growth-promoting, soil-adapted, and entomopathogenic species to cabbage maggots and were assayed in in vitro and soil laboratory bioassays. In the in vitro tests, the conidial suspensions were applied directly to cabbage maggot eggs. The soil tests mimicked pathways of natural exposure of various insect life stages to the fungal strains. Conidial concentrations used in soil tests were comparable to economic rates for in-furrow application. The following fungi were tested: Trichoderma atroviride P. Karst. (2 isolates), Trichoderma koningiopsis Samuels, C. Suárez & H.C. Evans (1), Trichoderma gamsii Samuels & Druzhin. (3), Beauveria brongniartii (Saccardo) Petch (1), Beauveria bassiana (Balsamo-Crivelli) Vuillemin (2), Metarhizium robertsii J.F. Bisch., Rehner & Humber (1), Metarhizium anisopliae (Metschn.) Sorokin (4), Purpureocillium lilacinum (Thom) Luangsa-ard, Houbraken, Hywel-Jones & Samson (2), and Clonostachys solani f. nigrovirens (J.F.H. Beyma) Schroers (2). Abbott's corrected mortality in the in vitro tests ranged from 0.0 +/- 18.9 to 47.6 +/- 9.0% and in the soil test from 2.4 +/- 13.0 to 68.2 +/- 21.5%. Seven isolates (B. bassiana [isolate 1174], C. solani [1828], M. anisopliae [1154 and 1868], T. atroviride [1872], T. koningiopsis [1874], and T. gamsii [1876]) caused significant cabbage maggot mortality in either in vitro or soil tests. The importance of fungal ecology as a criterion during the screening of potential biological control agents is discussed.

  10. evaluation of indigenous fungal isolates and metarhizium anisopliae ...

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    native fungal isolates against the lesser wax moth and assessing non target effect of one isolate of. Beauveria (IITA 18) and five ... widely distributed and devastating insect pest to ..... Non-Target. Invertebrates of Fungal Biocontrol Agents, PP.

  11. Isolation and identification of Metarhizium anisopliae from Chilo ...

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    2012-04-12

    Apr 12, 2012 ... Control of Tropical Crops Diseases and Insect Pests), Danzhou 571737, Hainan Province, ... well as the application of frequency vibration-killing lamp ..... Bioinsecticidal Strain of Pseudomonas fluorescens Active against the.

  12. Variability in the production of extracellular enzymes by entomopathogenic fungi grown on different substrates

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    Elio Gomes Fernandes

    2012-06-01

    Full Text Available Entomopathogenic fungi are important controllers of pest-insects populations in agricultural production systems and in natural environment. These fungi have enzymatic machinery which involve since the recognition and adherence of spores in their hosts culminating with infection and death of these insects. The main objective of this study was to analyzed extracellular enzyme production of the fungi strains Beauveria bassiana, Metarhizium anisopliae and Paecilomyces sp when cultured on substrates. These fungi were grown in minimal media containing specific substrates for the analysis of different enzymes such as amylases, cellulases, esterases, lipases, proteases (gelatin and caseinase, pectinases and cuticles of Musca domestica larvae and adults. All the assays were performed with and without the presence of dextrose in the culture media. The quantification of enzyme activity was performed by the ratio of halo / colony (H/C and the results subjected to variance analysis level of 5% (ANOVA followed by post-Tukey test. All strains were positive for lipase and also they showed a high significant enzyme production for gelatin at concentrations of 4 and 1%. B. bassiana and Paecilomyces sp. were positive for amylase, pectinase and caseinase, and only Paecilomyces sp. showed cellulase activity.

  13. [Species diversity and temporal niche of entomopathogenic fungi in the extensively managed tea plantation soil].

    Science.gov (United States)

    Guo, Xian-Jian; Shen, Wan-Fang; Liu, Yu-Jun; Chen, Ming-Jun

    2014-11-01

    The species diversity and temporal niche of entomopathogenic fungi community in the rhizosphere soil collected from the extensively managed Huangshan fuzz tip tea plantation were investigated. A total of 140 soil samples were collected at the location of Tangkou Town, Huangshan of Anhui Province during August, 2012 to June, 2013, and totally 1041 fungal isolates were obtained on selective medium with soil dilution plating. The results showed that the entomopathogenic fungi community in the tea plantation soil was diverse with 13 species in 6 genera. Purpureocillium lilacinum (309 strains), Beauveria bassiana (255 strains), and Metarhizium anisopliae (101 strains) were the dominant species accounting for 29.7%, 24.5% and 9.7% of the relative frequency, respectively. P. lilacinum had the widest temporal niche breadth among these dominant entomopathogenic fungi from the tea plantation soil, while B. bassiana had the narrowest. Among the entomopathogenic fungi, B. bassiana and B. brongniartii had the biggest temporal niche overlap of 1.965, while Isaria javanicus and B. bassiana had the smallest of 0.374.

  14. Naturally occurring entomopathogenic fungi infecting stored grain insect species in Punjab, Pakistan.

    Science.gov (United States)

    Wakil, Waqas; Usman Ghazanfar, Muhammad; Yasin, Muhammad

    2014-01-01

    The occurrence of entomopathogenic fungi isolated from stored grain insect pests sampled from various geographical regions of Punjab, Pakistan, was investigated. In total, 25,720 insects from six different species were evaluated, and 195 isolates from 24 different fungal species were recovered. These included the Ascomycetes Beauveria bassiana sensu lato (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae), Metarhizium anisopliae sensu lato (Metschnikoff) Sorokin (Hypocreales: Clavicipitaceae), Purpureocillium lilacinum (Thorn) Samson (Hypocreales: Ophiocordycipitaceae), and Lecanicillium attenuatum (Zare and W. Gams) (Hypocreales: Clavicipitaceae). The cadavers of red flour beetle Tribolium castaneum (Herbst.) (Coleoptera: Tenebrionidae) were significantly infected with the fungi followed by rice weevil Sitophilus oryzae (L.) (Coleoptera: Curculionidae), lesser grain borer Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae), rusty grain beetle Cryptolestes ferrugineus (Stephens) (Coleoptera: Cucujidae), and cowpea weevil Callosobruchus maculatus (F.) (Coleoptera: Bruchidae); however, the least were recovered from khapra beetle Trogoderma granarium (Everts) (Coleoptera: Dermestidae). The geographical attributes (altitude, longitude, and latitude) greatly influenced the occurrence of entomopathogenic fungi with highest number of isolates found from >400 (m) altitude, 33°-34' N latitude, and 73°-74' E longitude. The findings of the current surveys clearly indicated that the entomopathogenic fungi are widely distributed in the insect cadavers, which may later be used in successful Integrated Pest Management programs.

  15. Effects of carbon concentration and carbon to nitrogen ratio on the growth and sporulation of several biocontrol fungi.

    Science.gov (United States)

    Gao, Li; Sun, Man H; Liu, Xing Z; Che, Yong S

    2007-01-01

    Effects of carbon concentration and carbon to nitrogen (C:N) ratio on six biocontrol fungal strains are reported in this paper. All fungal strains had extensive growth on the media supplemented with 6-12 gl(-1) carbon and C:N ratios from 10:1 to 80:1, and differed in nutrient requirements for sporulation. Except for the two strains of Paecilomyces lilacinus, all selected fungi attained the highest spore yields at a C:N ratio of 160:1 when the carbon concentration was 12 gl(-1) for Metarhizium anisopliae SQZ-1-21, 6 gl(-1) for M. anisopliae RS-4-1 and Trichoderma viride TV-1, and 8 gl(-1) for Lecanicillium lecanii CA-1-G. The optimal conditions for P. lilacinus sporulation were 8 gl(-1) carbon with a C:N ratio of 10:1 for M-14 and 12 gl(-1) carbon with a C:N ratio of 20:1 for IPC-P, respectively. The results indicated that the influence of carbon concentration and C:N ratio on fungal growth and sporulation is strain dependent; therefore, consideration for the complexity of nutrient requirements is essential for improving yields of fungal biocontrol agents.

  16. Phylogenic analysis of adhesion related genes Mad1 revealed a positive selection for the evolution of trapping devices of nematode-trapping fungi.

    Science.gov (United States)

    Li, Juan; Liu, Yue; Zhu, Hongyan; Zhang, Ke-Qin

    2016-03-04

    Adhesions, the major components of the extracellular fibrillar polymers which accumulate on the outer surface of adhesive traps of nematode-trapping fungi, are thought to have played important roles during the evolution of trapping devices. Phylogenetic analyses based on the genes related to adhesive materials can be of great importance for understanding the evolution of trapping devices. Recently, AoMad1, one homologous gene of the entomopathogenic fungus Metarhizium anisopliae cell wall protein MAD1, has been functionally characterized as involved in the production of adhesions in the nematode-trapping fungus Arthrobotrys oligospora. In this study, we cloned Mad1 homologous genes from nematode-trapping fungi with various trapping devices. Phylogenetic analyses suggested that species which formed nonadhesive constricting ring (CR) traps more basally placed and species with adhesive traps evolved along two lineages. Likelihood ratio tests (LRT) revealed that significant positive selective pressure likely acted on the ancestral trapping devices including both adhesive and mechanical traps, indicating that the Mad1 genes likely played important roles during the evolution of nematode-trapping fungi. Our study provides new insights into the evolution of trapping devices of nematode-trapping fungi and also contributes to understanding the importance of adhesions during the evolution of nematode-trapping fungi.

  17. Further Screening of Entomopathogenic Fungi and Nematodes as Control Agents for Drosophila suzukii

    Science.gov (United States)

    Cuthbertson, Andrew G. S.; Audsley, Neil

    2016-01-01

    Drosophila suzukii populations remain low in the UK. To date, there have been no reports of widespread damage. Previous research demonstrated that various species of entomopathogenic fungi and nematodes could potentially suppress D. suzukii population development under laboratory trials. However, none of the given species was concluded to be specifically efficient in suppressing D. suzukii. Therefore, there is a need to screen further species to determine their efficacy. The following entomopathogenic agents were evaluated for their potential to act as control agents for D. suzukii: Metarhizium anisopliae; Isaria fumosorosea; a non-commercial coded fungal product (Coded B); Steinernema feltiae, S. carpocapsae, S. kraussei and Heterorhabditis bacteriophora. The fungi were screened for efficacy against the fly on fruit while the nematodes were evaluated for the potential to be applied as soil drenches targeting larvae and pupal life-stages. All three fungi species screened reduced D. suzukii populations developing from infested berries. Isaria fumosorosea significantly (p < 0.001) reduced population development of D. suzukii from infested berries. All nematodes significantly reduced adult emergence from pupal cases compared to the water control. Larvae proved more susceptible to nematode infection. Heterorhabditis bacteriophora proved the best from the four nematodes investigated; readily emerging from punctured larvae and causing 95% mortality. The potential of the entomopathogens to suppress D. suzukii populations is discussed. PMID:27294962

  18. Further Screening of Entomopathogenic Fungi and Nematodes as Control Agents for Drosophila suzukii

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    Andrew G. S. Cuthbertson

    2016-06-01

    Full Text Available Drosophila suzukii populations remain low in the UK. To date, there have been no reports of widespread damage. Previous research demonstrated that various species of entomopathogenic fungi and nematodes could potentially suppress D. suzukii population development under laboratory trials. However, none of the given species was concluded to be specifically efficient in suppressing D. suzukii. Therefore, there is a need to screen further species to determine their efficacy. The following entomopathogenic agents were evaluated for their potential to act as control agents for D. suzukii: Metarhizium anisopliae; Isaria fumosorosea; a non-commercial coded fungal product (Coded B; Steinernema feltiae, S. carpocapsae, S. kraussei and Heterorhabditis bacteriophora. The fungi were screened for efficacy against the fly on fruit while the nematodes were evaluated for the potential to be applied as soil drenches targeting larvae and pupal life-stages. All three fungi species screened reduced D. suzukii populations developing from infested berries. Isaria fumosorosea significantly (p < 0.001 reduced population development of D. suzukii from infested berries. All nematodes significantly reduced adult emergence from pupal cases compared to the water control. Larvae proved more susceptible to nematode infection. Heterorhabditis bacteriophora proved the best from the four nematodes investigated; readily emerging from punctured larvae and causing 95% mortality. The potential of the entomopathogens to suppress D. suzukii populations is discussed.

  19. Further Screening of Entomopathogenic Fungi and Nematodes as Control Agents for Drosophila suzukii.

    Science.gov (United States)

    Cuthbertson, Andrew G S; Audsley, Neil

    2016-06-09

    Drosophila suzukii populations remain low in the UK. To date, there have been no reports of widespread damage. Previous research demonstrated that various species of entomopathogenic fungi and nematodes could potentially suppress D. suzukii population development under laboratory trials. However, none of the given species was concluded to be specifically efficient in suppressing D. suzukii. Therefore, there is a need to screen further species to determine their efficacy. The following entomopathogenic agents were evaluated for their potential to act as control agents for D. suzukii: Metarhizium anisopliae; Isaria fumosorosea; a non-commercial coded fungal product (Coded B); Steinernema feltiae, S. carpocapsae, S. kraussei and Heterorhabditis bacteriophora. The fungi were screened for efficacy against the fly on fruit while the nematodes were evaluated for the potential to be applied as soil drenches targeting larvae and pupal life-stages. All three fungi species screened reduced D. suzukii populations developing from infested berries. Isaria fumosorosea significantly (p entomopathogens to suppress D. suzukii populations is discussed.

  20. Molecular identification of Metarhizium strains isolated from soil in Anhui%安徽省土壤绿僵菌分离株的分子鉴定

    Institute of Scientific and Technical Information of China (English)

    周玉宝; 聂勇; 黄勃

    2011-01-01

    目的 进一步验证ITS序列的系统发育分析可为绿僵菌属种的鉴定提供重要的参考依据.方法 对分离自安徽土壤的13株绿僵菌菌株的内转录间隔区(ITS)片段进行PCR扩增和序列测定,采用Blast方法 将测序结果 在GenBank中进行同源搜索,依据邻接法构建获得与其相关菌株的ITS序列系统发育树.结果 供试菌株分别位于系统发育树的3个分支上,分支Ⅰ包括8个菌株和金龟子绿僵菌小孢变种,1个菌株和金龟子绿僵菌鳞鳃金龟变种形成分支Ⅲ,另外4个菌株和黄绿绿僵菌棉蚜变种聚为分支X.结论 结合同源比较的数据,将这8个、4个和1个绿僵菌菌株分别鉴定为金龟子绿僵菌小孢变种、黄绿绿僵菌棉蚜变种和金龟子绿僵菌鳞鳃金龟变种.%Objective To further manifest that the phylogenetic analysis of ITS sequence could provide an important reference basis for genus identification of Metarhizium. Method The internal transcribed spacer (ITS) sequences of 13 Metarhizium strains isolated from Anhui soil was amplified and sequenced. We searched for homology using the BLAST programs available at the web site of the National Center for Biotechnology Information, and the phylogenetic tree for the genus Metarhizium was constructed by Neighbor-Joining method. Result The tested strains were located in 3 clades of the phylogenetic tree. 8 Metarhizium strains and the Metarhizium anisopliae var. Anisopliae type were classified into clade I, 1 strain and the M. Anisopliae var. Lepidiotum type were grouped into clade Ⅲ, and 4 strains and the M. Jlavoviride var. Pemphigum type were classified into clade X. -Conclusion Combined with the homologous comparative data, the eight strains, the one strain and the four strains are identified as Metarhizium anisopliae var. Anisopliae, M. Anisopliae var. Lepidiotum and M. Jlavoviride var. Pemphigum, respectively.

  1. Biocontrol of the Brown-banded Cockroach, Supella longipalpa F. (Blattaria: Blattellidae, with Entomopathogenic Fungus, Metharhizium anisopliae

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    Mona Sharififard

    2016-01-01

    Full Text Available Background: Considering to the high distribution of cockroaches as urban pests, the efficacy of different formula­tions of Metarhizium anisopliae strain Iran 437C were assessed against the brown-banded cockroach, Supella longi­palpa F. under laboratory and field conditions.Methods: Metarhizium anisopliae isolates were screened with immersing adults of the brown-banded cockroachs in aqueous suspension of 108 conidia ml-1 followed by surface or bait treated with different doses of the most virulent isolate against the nymphs. Then formulations of conidia oil-in-water were examined versus cockroach nymphs us­ing different plant oils and paraffin. Then they were evaluated and compared with aqueous suspension and control group. On a large-scale, the sunflower oil-in-water formulation of conidia was sprayed at houses using a hand sprayer.Results: Metarhizium anisopliae IRAN 437C was the most virulent isolate against the brown-banded cockroach, causing 100% mortality in adults at seven days post-exposure. Inoculated bait with this isolate was not enough path­ogenic against the cockroach even at two weeks after treatment. Treated surface with conidia as aqueous suspension or oil-in- water formulation was more effective than the bait formulation against the cockroach caused 39.4–97.2% mortality compared with 2.5% mortality in control group after two days. Spraying the conidia formulated with sun­flower oil was an effective formulation causing 76.1% reduction in the cockroach density on the third day post treatment in the houses.Conclusion: The oil-in-water formulation of M. anisopliae IRAN 437C could be recommended as a promising al­ternative for cockroach control.  

  2. ACTIVITY OF SOME BRAZILIAN ISOLATES OF ENTOMOPATHOGENIC FUNGI AGAINST THE POULTRY RED MITE DERMANYSSUS GALLINAE DE GEER (ACARI: DERMANYSSIDAE

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    CR Kasburg

    Full Text Available ABSTRACT Poultry red mite Dermanyssus gallinae is a cosmopolitan and hematophagous species commonly found in layer houses around the world. Poultry mite infestations may cause anemia, stress, low body weight and egg production, and mortality. Mite control is typically based on chemical products, but they are not effective and leave residues in eggs; therefore, alternative control methods, such as entomopathogenic fungi, need to researched. This study aimed at evaluating, in the laboratory, the activity of Brazilian isolates of entomopathogenic fungi against D. gallinae. The mites were collected from a commercial layer house and were sprayed with conidial suspensions (1 × 108 conidia/mL of five isolates of Beauveria bassiana and Metarhizium anisopliae. All tested isolates were pathogenic for the red mite, with confirmed mortality ranging from 22.9 to 52.4%. This demonstrate the potential of the tested entomopathogenic fungi isolates for mite control, and reinforces the need for further studies with other isolates, application strategies, and with fungal formulations.

  3. Fungi associated with the hemlock woolly adelgid, Adelges tsugae, and assessment of entomopathogenic isolates for management.

    Science.gov (United States)

    Reid, W R; Parker, B L; Gouli, S Y; Skinner, M; Gouli, V V; Teillon, H B

    2010-01-01

    Fungi associated with the hemlock wooly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), were collected throughout the eastern USA and southern China. Twenty fungal genera were identified, as were 79 entomopathogenic isolates, including: Lecanicillium lecanii (Zimmermann) (Hypocreales: Insertae sedis), Isaria farinosa (Holm: Fries.) (Cordycipitaceae), Beauveria bassiana (Balasamo) (Hyphomycetes), and Fusarium spp (Nectriaceae). The remaining fungal genera associated with insect cadavers were similar for both the USA and China collections, although the abundance of Acremonium (Hypocreaceae) was greater in China. The entomopathogenic isolates were assayed for efficacy against Myzus persicae (Sulzer) (Homoptera: Aphididae) and yielded mortality ranging from 3 to 92%. Ten isolates demonstrating the highest efficacy were further assessed for efficacy against field-collected A. tsugae under laboratory conditions. Overall, two B. bassiana, one L. lecanii, and a strain of Metarhizium anisopliae (Metchnikoff) (Hypocreales: Clavicipitaceae), demonstrated significantly higher efficacy against A. tsugae than the others. Isolates were further evaluated for conidial production, germination rate and colony growth at four temperatures representative of field conditions. All isolates were determined to be mesophiles with optimal temperature between 25-30 degrees C. In general, conidial production increased with temperature, though two I. farinosa produced significantly more conidia at cooler temperatures. When efficacy values were compared with conidial production and temperature tolerances, Agricultural Research Service Collection of Entomopathogenic Fungi (ARSEF) 1080, 5170, and 5798 had characteristics comparable to the industrial B. bassiana strain GHA.

  4. Influence of substrate and relative humidity on the efficacy of three entomopathogenic fungi for the hide beetle, Dermestes maculatus

    Science.gov (United States)

    Dermestes maculatus is carrion feeder that is also a pest of poultry houses, museums, silkworm culture, and many stored foods. The Hypocreales, Beauveria bassiana, Metarhizium anisopliae, and Isaria fumosorosea, were tested for efficacy against D. maculatus larvae on concrete, plastic, leather, and ...

  5. Screening of Fungi for Biological Control of a Triatomine Vector of Chagas Disease: Temperature and Trypanosome Infection as Factors

    Science.gov (United States)

    Garcia, Aline R. M.; Rocha, Adriana de Paula; Moreira, Camila C.; Rocha, Silma L.; Guarneri, Alessandra A.; Elliot, Simon L.

    2016-01-01

    Entomopathogenic fungi have been investigated as an alternative tool for controlling various insects, including triatomine vectors of the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. Here we tested the pathogenicity and virulence of ten isolates of the fungi Metarhizium spp. and Beauveria bassiana against Rhodnius prolixus and found all of the isolates to be virulent. We used two isolates (URPE-11 Metarhizium anisopliae and ENT-1 Beauveria bassiana) for further screening based on their prolific sporulation in vitro (an important property of fungal biopesticides). We characterized their virulences in a dose-response experiment and then examined virulence across a range of temperatures (21, 23, 27 and 30°C). We found isolate ENT-1 to maintain higher levels of virulence over these temperatures than URPE-11. We therefore used B. bassiana ENT-1 in the final experiment in which we examined the survival of insects parasitized with T. cruzi and then infected with this fungus (once again over a range of temperatures). Contrary to our expectations, the survival of insects challenged with the pathogenic fungus was greater when they had previously been infected with the parasite T. cruzi than when they had not (independent of temperature). We discuss these results in terms of aspects of the biologies of the three organisms. In practical terms, we concluded that, while we have fungal isolates of potential interest for development as biopesticides against R. prolixus, we have identified what could be a critical problem for this biological tool: the parasite T. cruzi appears to confer a measure of resistance to the insect against the potential biopesticide agent so use of this fungus as a biopesticide could lead to selection for vector competence. PMID:27855217

  6. Potential for entomopathogenic fungi to control Triatoma dimidiata (Hemiptera: Reduviidae, a vector of Chagas disease in Mexico

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    María Guadalupe Vázquez-Martínez

    2014-12-01

    Full Text Available Introduction The use of entomopathogenic fungi to control disease vectors has become relevant because traditional chemical control methods have caused damage to the environment and led to the development of resistance among vectors. Thus, this study assessed the pathogenicity of entomopathogenic fungi in Triatoma dimidiata. Methods Preparations of 108 conidia/ml of Gliocladium virens, Talaromyces flavus, Beauveria bassiana and Metarhizium anisopliae were applied topically on T. dimidiata nymphs and adults. Controls were treated with the 0.0001% Tween-80 vehicle. Mortality was evaluated and recorded daily for 30 days. The concentration required to kill 50% of T. dimidiata (LC50 was then calculated for the most pathogenic isolate. Results Pathogenicity in adults was similar among B. bassiana, G. virens and T. flavus (p>0.05 and differed from that in triatomine nymphs (p=0.009. The most entomopathogenic strains in adult triatomines were B. bassiana and G. virens, which both caused 100% mortality. In nymphs, the most entomopathogenic strain was B. bassiana, followed by G. virens. The native strain with the highest pathogenicity was G. virens, for which the LC50 for T. dimidiata nymphs was 1.98 x108 conidia/ml at 13 days after inoculation. Conclusions Beauveria bassiana and G. virens showed entomopathogenic potential in T. dimidiata nymphs and adults. However, the native G. virens strain presents a higher probability of success in the field, and G. virens should thus be considered a potential candidate for the biological control of triatomine Chagas disease vectors.

  7. EFFECTS OF SOME BIOINSECTICIDES AND ENTOMOPATHOGENIC FUNGI ON COLORADO POTATO BEETLE (LEPTINOTARSA DECEMLINEATA L.).

    Science.gov (United States)

    Öztürk, H E; Güven, Ö; Karaca, I

    2015-01-01

    In this study, biological activity of entomopathogenic fungi (4 strains) isolated from the Colorado potato beetle and the commercial biopesticides containing entomopathogenic fungi; Priority® (Paecilomyces fumosoroseus), Nibortem® (Verticillium lecanii), Nostalgist® (Beauveria bassiana), Bio-Magic* (Metarhizium anisopliae), Bio-Nematon* (Paeciliomyces sp.) and plant extracts; Nimbedicine EC* (Azadiractin) were determined against Leptinotarsa decemlineata under laboratory conditions. An Imidacloprid active ingredient commercial insecticide was also used to compare the insecticidal activity and distilled water was used as control. The biological control agents were applied to 2nd-3rd larval instars, 4th larval instars and adults with spray and leaf dipping methods. Single concentration (10⁸ conidia/mL⁻¹) of entomopathogenic fungi and recommended dose of bioinsecticides were prepared for application. The number of dead insects were determined at 3, 5, and 7 days after applications. Experiments were conducted at 25 ±1° C and 60% ± 5 relative humidity with 16:8 h light: dark conditions. Entomopathogenic fungi and bioinsecticides were found to be more effective on larval stage than 4th larval instars and adults. In spray methods, Bio-Magic®, Nibortem®, and Nostalgist® caused 96.4%, 92.9% and 82.1% mortality on 2nd larval instars and 20%, 36.7% and 33.3% mortality on adults, respectively. All local fungal isolates (B. bassiana) applied on 2nd and 4th larval instars caused 100% mortality. Adults showed 58.6-86.2% mortality.

  8. Fungal Entomopathogens in the Rhizosphere

    Science.gov (United States)

    Entomopathogenic fungi are found in a wide variety of fungal groups. The order Hypocreales contains the largest number of entomogenous fungi, including two of the most widely studied, Beauveria bassiana (Balsamo) Vuillemin (Hypocreales: Clavicipitaceae) and Metarhizium anisopliae (Metchnikoff) Sorok...

  9. Entomopathogenic fungi isolates to control the borer of yerba mate (Hedypathes betulinus Kluger (Coleoptera; CerambycidaeIsolados de fungos entomopatogênicos visando ao controle da broca da erva-mate (Hedypathes betulinus Kluger (Coleoptera; Cerambycidae

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    André Luis Pereira Fanti

    2013-09-01

    Full Text Available Yerba mate is an important crop of southern Brazil, Argentina and Paraguay, with socio-economic and environmental importance. Due the lack of native herbals, yerba mate has become a monoculture, and as a consequence, insects have become pests, and the borer, Hedypathes betulinus, one of the most important. In order to improve biological control in the crop, this work aims to select in laboratory the most virulent, of 32 isolates of the entomopathogenic fungi Beauveria bassiana (Bals. Vuill. and 18 isolates of Metarhizium anisopliae (Metsch. Sorok. Initially, isolates virolence was evaluated by immersing the adults borers in a fungal suspension, containing 1×108 conidia mL-1, and after, those caused highest confirmed mortality were selected for the second step, wherein it was evaluated the virulence, vegetative growth and conidia production in culture medium, rice and borer bodies. Three isolates of B. bassiana (UNIOESTE 4, UNIOESTE 52 and UNIOESTE 64 and one of M. anisopliae (IBCB 352 reached confirmed mortality equal or superior to 90%, these isolates were selected for the second step, together with the standard isolated GC 716 (B. bassiana. The isolated IBCB 352 of M. anisopliae, despite being the only of this species to be selected for the second step, promoted the high confirmed mortality and conidiogenesis in adults borer, indicating a high inoculums production i, being nominated for field evaluations. A erva-mate é uma cultura importante da região sul do Brasil, Argentina e Paraguai, tendo grande importância sócio-econômica e ambiental. Com a escassez de ervais nativos, a cultura da erva-mate passou a ser cultivada em sistema de monocultivo e como consequência, alguns insetos tornaram-se pragas, sendo a broca, Hedypathes betulinus, muito importante. A fim de incrementar o controle biológico na cultura, o objetivo deste trabalho foi selecionar em laboratório os isolados mais virulentos de 32 isolados do fungo entomopatog

  10. Effects of transgenic Cry1Ac cotton on entomopathogenic fungi in soil%转Cry1Ac抗虫棉对土壤虫生真菌群落的影响

    Institute of Scientific and Technical Information of China (English)

    谢明; 张艳军; 吴刚

    2012-01-01

    The species diversity and amount of soil fungi around the roots of the transgenic GK-12 cotton variety containing the CrylAc gene was compared to that of the non-transgenic SM-3 variety. There was no significant difference in soil fungal species between GK-12 and SM-3 plants, with the fungi Beauveria bassiana and Metarhizium anisopliae found in the root area of both varieties. There were, however, significant differences in the soil fungal species associated with these two varieties during the boll forming and boll opening periods. There were no differences in species or quantity of fungi found at different distances (5 cm, 15 cm and 25 cm) from the roots of these cotton varieties. These results indicate that the presence of the CrylAc gene had no significant effect on fungal species and quantity.%利用大蜡螟Galleria mellonella L.诱菌法对转Cry1Ac抗虫棉GK-12及其亲本棉花SM-3的根系区域土壤的虫生真菌种类和数量进行监测,评价转Cry1Ac基因对棉田土壤虫生真菌的影响.实验结果显示:检测到的虫生真菌种类,在转基因棉GK-12及其亲本棉SM-3之间无显著差异,均为球孢白僵菌(Beauveria bassiana)和金龟子绿僵菌(Metarhizium anisopliae);在棉花开花初期、盛花期、结铃期或吐絮期,土壤虫生真菌数量在抗虫棉GK-12和亲本SM-3之间存在显著的差异,其他生育期无显著差异;在距棉花主根不同距离(5cm\\15cm\\25cm)的区域,土壤虫生真菌数量和种类在抗虫棉GK-12和亲本SM-3之间无显著差异.

  11. Laboratory and field evaluation of entomopathogenic fungi for the control of amitraz-resistant and susceptible strains of Rhipicephalus decoloratus.

    Science.gov (United States)

    Murigu, Mercy M; Nana, Paulin; Waruiru, Robert M; Nga'nga', Chege J; Ekesi, Sunday; Maniania, Nguya K

    2016-07-30

    Rhipicephalus decoloratus causes serious economic losses in cattle industry every year in East Africa. Biological control using entomopathogenic fungi is seen as a promising alternative to chemical acaricides being used for their control. The pathogenicity of Metarhizium anisopliae and of Beauveria bassiana isolates was tested in the laboratory against amitraz-resistant and amitraz-susceptible strains of R. decoloratus. Unfed larvae were sprayed with conidial suspensions of 1×10(9) conidia ml(-1). Fungal isolates were pathogenic to R. decoloratus larvae, causing mortality of between 10.0 and 100% and between 12.1 and 100% of amitraz-susceptible and amitraz-resistant strains, respectively. The LT50 values of selected fungal isolates varied between 2.6-4.2days in amitraz-susceptible strain and between 2.8-3.9days in amitraz-resistant strain. The LC50 values varied between 0.4±0.1 and 200.0±60×10(3) conidia ml(-1) and between 0.1±0.1 and 200.0±31.0×10(3) conidia ml(-1) in amitraz-susceptible and amitraz-resistant strains, respectively. Metarhizium anisopliae isolate ICIPE 7 outperformed the other isolates and was selected for compatibility study with amitraz and field trial. ICIPE 7 was compatible with amitraz. In the field, four treatments including control, ICIPE 7 alone, amitraz alone and ICIPE 7/amitraz were applied on cattle. All the treatments significantly reduced the number of ticks on all the sampling dates: day 7 (F3,8=3.917; P=0.0284), day 14 (F3,8=9.090; P=0.0275), day 21 (F3,8=37.971; P=0.0001) and day 28 (F3,8=8.170; P=0.0016) compared to the control. Results of the present study indicate that ICIPE 7 can be used for the management of amitraz-resistant strain of R. decoloratus.

  12. Infection of the malaria mosquito, Anopheles gambiae, with two species of entomopathogenic fungi: effects of concentration, co-formulation, exposure time and persistence

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    Lwetoijera Dickson W

    2009-12-01

    Full Text Available Abstract Background Entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana isolates have been shown to infect and reduce the survival of mosquito vectors. Methods Here four different bioassays were conducted to study the effect of conidia concentration, co-formulation, exposure time and persistence of the isolates M. anisopliae ICIPE-30 and B. bassiana I93-925 on infection and survival rates of female Anopheles gambiae sensu stricto. Test concentrations and exposure times ranged between 1 × 107 - 4 × 1010 conidia m-2 and 15 min - 6 h. In co-formulations, 2 × 1010 conidia m-2 of both fungus isolates were mixed at ratios of 4:1, 2:1, 1:1,1:0, 0:1, 1:2 and 1:4. To determine persistence, mosquitoes were exposed to surfaces treated 1, 14 or 28 d previously, with conidia concentrations of 2 × 109, 2 × 1010 or 4 × 1010. Results Mosquito survival varied with conidia concentration; 2 × 1010 conidia m-2 was the concentration above which no further reductions in survival were detectable for both isolates of fungus. The survival of mosquitoes exposed to single and co-formulated treatments was similar and no synergistic or additive effects were observed. Mosquitoes were infected within 30 min and longer exposure times did not result in a more rapid killing effect. Fifteen min exposure still achieved considerable mortality rates (100% mortality by 14 d of mosquitoes, but at lower speed than with 30 min exposure (100% mortality by 9 d. Conidia remained infective up to 28 d post-application but higher concentrations did not increase persistence. Conclusion Both fungus isolates are effective and persistent at low concentrations and short exposure times.

  13. Multiplexed microsatellite markers for seven Metarhizium species

    Science.gov (United States)

    Cross-species transferability of 41 previously published simple sequence repeat (SSR) markers was assessed for 11 species of the entomopathogenic fungus Metarhizium. A collection of 65 Metarhizium isolates including all 54 used in a recent phylogenetic revision of the genus were characterized. Betwe...

  14. Diversity of rhizosphere associated entomopathogenic fungi of perennial herbs, shrubs and coniferous trees.

    Science.gov (United States)

    Fisher, Joanna J; Rehner, Stephen A; Bruck, Denny J

    2011-02-01

    Understanding habitat selection of fungal entomopathogens is critical to improve the efficacy, persistence and cost of these fungi as microbial insecticides. This study sought to determine the prevalence of Metarhizium and Beauveria spp. isolated from the rhizosphere of strawberry, blueberry, grape and Christmas tree crops in the Willamette Valley of Oregon. Entomopathogenic fungi were assigned to thirteen species based on molecular phylogenetic criteria. Four species of Metarhizium were isolated including Metarhizium brunneum, Metarhizium guizhouense, Metarhizium robertsii, and Metarhizium flavoviride var. pemphigi. Nine Beauveria species were isolated including, Beauveria brongniartii, an undescribed species referred to as Clade C and seven phylogenetic species of Beauveria bassiana. Strawberries and blueberries were significantly associated with M. brunneum and Christmas trees with M. guizhouense and M. robertsii. Grapes were significantly associated with B. bassiana phylogenetic species Bbas-16. All of the Metarhizium isolates screened were pathogenic to Otiorhynchus sulcatus larvae in laboratory bioassays but only M. brunneum and M. robertsii caused significant levels of infection. The study results suggest that certain species of Metarhizium and Beauveria are significantly associated with the strawberry, blueberry and Christmas tree rhizosphere and could potentially provide better control of O. sulcatus.

  15. Isolation of endosymbionts from Ipomoea carnea and Swainsona canescens that produce swainsonine

    Science.gov (United States)

    Fungi including Metarhizium anisopliae (Clavicipitaceae), Rhizoctonia leguminicola (Ceratobasidiaceae), and Undifilum (Pleosporaceae), an endophyte found in the plant genera Astragalus and Oxytropis (Fabaceae) have been reported to be responsible for the production of swainsonine. Based upon the ass...

  16. Behavioral and electrophysiological responses of Coptotermes formosanus Shiraki towards entomopathogenic fungal volatiles

    Science.gov (United States)

    Termites adjust their response to entomopathogenic fungi according to the profile of the fungal volatile organic compounds (VOCs). This study first demonstrated the pathogenicity of Metarhizium anisopliae, Beauveria bassiana and Isaria fumosorosea (=Paecilomyces fumosoroseus) towards the Formosan s...

  17. Advances in microbial insect control in horticultural ecosystems

    Science.gov (United States)

    The use of microbial organisms as biological control agents has progressed significantly since Metschnikoff launched the first attempt at microbial insect control with Metarhizium anisopliae in 1879. Following the lead of Metschnikoff, entomopathogenic nematodes, fungi, bacteria and viruses have b...

  18. Characterization and activity of entomopathogenic fungi isolates against “paraguay tea ampul” (Gyropsylla spegazziniana (Lizer & Trelles (Hemiptera: Psyllidae

    Directory of Open Access Journals (Sweden)

    Marina Andressa Formentini

    2015-12-01

    Full Text Available Paraguay tea is a native crop of southern Brazil and is also socioeconomically important in Paraguay and Argentina. One of the main pests of this crop is Gyropsylla spegazziniana (Hemiptera: Psyllidae. As there are no registered insecticides for this pest in Brazil, the use of entomopathogenic fungi is an alternative method for its control. This study aimed to evaluate and characterize isolates of the entomopathogenic fungi Beauveria spp., Metarhizium anisopliae, Isaria spp. and Lecanicillium spp. on G. spegazziniana. For this purpose, 5th instar nymphs were transferred to Paraguay tea seedlings, followed by spraying of conidial suspensions (1 × 109 conidia mL-1 and then placing the seedlings in polyvinyl chloride (PVC cages and maintaining them in a temperature-controlled room (26 ± 1°C, 12h00 photophase, 60 ± 10% R.H.. Insect mortality was evaluated daily for 10 days, and the selected isolates were characterized according to their vegetative growth, conidial production in synthetic culture medium and rice and insecticidal activity as well as through molecular analyses, including sequencing the rDNA-ITS region and RAPD analysis. The genus Beauveria spp. was the most efficient, particularly the Unioeste 44 isolate, which caused the greatest total mortality of G. spegazziniana (81.7% and showed among the highest conidial production levels on rice, indicating a significant potential to be used in an integrated management program for this pest. Molecular analysis of the rDNA-ITS region allowed the isolates to be identified as B. bassiana and B. brongniartii, and RAPD markers were found to be associated with virulence.

  19. Presencia natural de hongos hyphomycetes en larvas invernantes de Diatraea saccharalis F. en caña de azúcar en Tucumán, Argentina Natural presence of hyphomycetes fungi in hibernating Diatraea saccharalis F. larvae in sugarcane in Tucumán, Argentina

    Directory of Open Access Journals (Sweden)

    Marta G. Yasem de Romero

    2008-07-01

    Full Text Available El "gusano perforador de la caña de azúcar" Diatraea saccharalis (Fabricius (Lepidoptera: Pyralidae es la plaga más importante del cultivo de caña de azúcar en Tucumán. La optimización del manejo de esta plaga requiere la adecuada combinación de diferentes recursos disponibles, incluso en la naturaleza. En el año 2005 se inició un relevamiento de los hongos entomopatógenos presentes en el agroecosistema cañero tucumano. El objetivo es detectar la presencia e identificar a los hongos entomopatógenos que actúan como agentes de mortalidad natural en larvas invernantes de D. saccharalis en caña de azúcar en Tucumán, Argentina. De las larvas analizadas muertas por hongos entomopatógenos, el 57,13% correspondió a Beauveria bassiana (Bals. Vuill., el 23,82 % Metarhizium anisopliae (Metsch. Sorokin, el 9,53% a Nomuraea rileyi (Farlow Samson y el 9,53% a Isaria sp. Persoon: Fries. Su presencia resultaría de importancia, ya que la densidad poblacional del gusano perforador podría estar regulada por los citados hyphomycetes.Sugarcane borer Diatraea saccharalis (Fabricius (Lepidoptera: Pyralidae is the most important pest affecting sugarcane crops in Tucumán. Optimizing its management entails combining different available resources, even natural ones, accurately. Since 2005, a survey of entomopathogenic fungi present in the local sugarcane agroecosystem has been under way. The aim is to detect and identify entomopathogenic fungi which cause natural death in hibernating D. saccharalis larvae affecting sugarcane in Tucumán, Argentina. By now, Beauveria bassiana (Bals. Vuill. (57.13%, Metarhizium anisopliae (Metsch. Sorokin (23.82%, Nomuraea rileyi (Farlow Samson (9.53% and Isaria sp. Persoon: Fries (9.53% have been found naturally in hibernating sugarcane borer larvae in sugarcane crops in the province of Tucumán. This finding may be relevant in suggesting that sugarcane borer population might be regulated by hyphomycetes.

  20. 感染水椰八角铁甲的绿僵菌的分子鉴定及致病力测定%Molecular identification and pathogenicity assay on Metarhizium against Octodonta nipae ( Coleoptera: Chrysomelidae)

    Institute of Scientific and Technical Information of China (English)

    徐利; 蓝江林; 侯有明; 陈玉森; 陈智雄; 翁章权

    2011-01-01

    在室内饲养的水椰八角铁甲Octodonta nipae(Maulik)种群中,发现有大量甲虫被病原菌感染致死.对死虫体表的病原真菌进行分离鉴定,并依据ITS序列分析鉴定,确定该病原真菌为金龟子绿僵菌小孢变种(Metarhizium anisopliae var.anisopliae).经室内致病力测定,接种浓度分别为1.0×105~1.0×1010cfu·mL-1孢子悬浮液,在接菌后10 d,成虫致病力方程为y=-4.992+5.623x,LC50对数剂量为7.721;幼虫致病力方程为y=-4.335+5.368 x,LC50对数剂量为6.420.结果表明金龟子绿僵菌小孢变种对水椰八角铁甲幼虫和成虫具有高的致病力,且浓度越高感染率越高,相同浓度条件下幼虫的感染率较成虫高.金龟子绿僵菌小孢变种在水椰八角铁甲生物防治中具有较强的应用潜力.%High adult and larval mortality due to infection with a pathogenic fungus was observed when rearing Octodonta nipae (Maulik) under laboratory conditions. ITS sequence analysis identified the fungus as Metarhizium anisopliae var.anisopliae. The effect of inoculating 3rd instar larvae and adults of O. nipae with six different concentrations of M.anisopliae var. anisopliae was investigated. Dose-response bioassays showed that M. anisopliae var. anisopliae had a high level of toxicity to both larvae and adults. At concentrations ranging from 1.0 × l05 cfu · mL-1to 1.0 × 1010 cfu · mL-1 ,the virulence equations on the 10th day after infection were y = - 4.335 + 5.368 x and y = - 4.992 + 5.623x for larvae and adults, respectively. The estimated logarithm of median lethal concentration was 6.420 and 7.721 for larvae and adults, respectively. These results indicate that M. anisopliae var. anisopliae was highly virulent against O. nipae larvae and adults and that the degree of virulence increased with increasing concentration. The infection rate was higher in larvae than in adults at the same concentration. These results indicate that M. anisopliae var. anisopliae could be a

  1. The use of entomopathogenic fungi for the control of parasitic mites, Psoroptes spp.

    Science.gov (United States)

    Smith, K E; Wall, R; French, N P

    2000-09-20

    In vitro trials were carried out to evaluate the potential of fungal pathogens as biological control agents of parasitic mites, Psoroptes ovis (Hering) (Acari: Psoroptidae) from rabbit hosts (synonym: Psoroptes cuniculi). The fungus Hirsutella thompsonii Fisher showed no pathogenicity. Metarhizium anisopoliae (Metschinkoff), however, showed a high level of pathogenicity; 3 days after exposure to fungal conidia all mites were dead and 6 days after exposure 60% of the dead adult females, 10% of the dead adult males and 30% of the dead female nymphs had fungal hyphae protruding from their cuticular surface. There was a significant effect of conidial concentration on the number of mite cadavers that displayed fungal infection. Exposure to between 1x10(4) and 1x10(6) conidiaml(-1) resulted in 2-25% of the mites being infected. Mean infection levels were highest, 71%, when the mites were exposed to 1x10(7) conidiaml(-1). Similarly, there was a significant effect of conidial concentration on the time taken for the mites to reach 50% levels of mortality (LT(50)) The mean LT(50) value was approximately 2.7 days, when the mites were exposed to a solution of 1x10(7) and 1x10(8) conidiaml(-1) which was significantly shorter than controls exposed to 0.03% Tween-80 solution only. There was no significant effect of passaging the fungus, either once or twice, through the host on the subsequent infectivity of M. anisopliae. The potential for use of entornopathogenic fungi for the control of parasitic mites, particularly in relation to sheep, is discussed.

  2. Storage conditions affect speed of germination in Beauveria bassiana and Metarhizium anisopliae conidia

    Science.gov (United States)

    The retention of high viability during storage is essential for effectiveness and thus market acceptance of fungus-based biopesticides. The length of adequate shelf-lives for mycoinsecticides is controversial, with proposed requirements varying from a few weeks to 18 months. Shelf-life determination...

  3. THE IDENTIFICATION AND CHARACTERIZATION OF AN LGE-INDUCING PROTEIN IN METARHIZIUM ANISOPLIAE EXTRACT

    Science.gov (United States)

    Molds are ubiquitous components of the indoor environment and have been associated with exacerbation of asthma as well as a number of other health effects. Their contribution to the induction of allergic asthma is less certain. Previously, we have shown that BALB/c mice exposed...

  4. Behavioural effects of fungal infection by Metarhizium anisopliae in adult malaria mosquitoes

    NARCIS (Netherlands)

    Ondiaka, S.N.

    2012-01-01

    Malaria remains a major global health problem with the burden of disease greatest in Sub-Saharan Africa. The strategies for malaria control differ throughout the world according to levels of endemicity and the magnitude of disease but the focus remains either to control malaria parasites or vectors.

  5. 76 FR 26194 - Metarhizium anisopliae Strain F52; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2011-05-06

    ... transferred from Earth BioSciences, Inc. to Novozymes Biologicals, Inc. (TAE-001 Technical Bioinsecticide, EPA Reg. No. 70127-7; Taenure Granular Bioinsecticide, EPA Reg. No. 70127-8; Tick-EX G, EPA Reg. No. 70127...

  6. Behavioural effects of fungal infection by Metarhizium anisopliae in adult malaria mosquitoes

    NARCIS (Netherlands)

    Ondiaka, S.N.

    2012-01-01

    Malaria remains a major global health problem with the burden of disease greatest in Sub-Saharan Africa. The strategies for malaria control differ throughout the world according to levels of endemicity and the magnitude of disease but the focus remains either to control malaria parasites or vectors.

  7. Efficacy of entomopathogenic fungi in combination with diatomaceous earth against Callosobruchus maculatus(Coleoptera: Bruchidae)%昆虫病原真菌与硅藻土混用对四纹豆象的毒力效果

    Institute of Scientific and Technical Information of China (English)

    Neda NABAEI; Ali MEHRVAR; Moosa SABER; Mohammad BAGHERI

    2012-01-01

    在室内条件下检测了白僵菌Beauveria bassiana (Balsamo) Vuillemin菌株IRAN-187C和金龟子绿僵菌Metarhizium anisopliae (Metschnikow) Sorokin菌株A-396分别与硅藻土(DE)制剂InsectoSec(R)混用对四纹豆象Callosobruchus maculatus成虫的毒力效果.结果表明:硅藻土与致死剂量和亚致死剂量的真菌分离物混用导致了四纹豆象成虫的高死亡率,且这些处理均缩短了真菌的致死中时(LT50).LC50的白僵菌B.bassiana菌株IRAN-187C和LC50的DE混用对四纹豆象的LT50 (122.2 h)最低,在95%置信限上与LC50的金龟子绿僵菌M.anisopliae菌株A-396与LC50的DE混用得到的LT50(128.2 h)相当,说明两种真菌分离物与DE混用对四纹豆象成虫都具有较好的毒力效果.用四纹豆象成虫检测3个温度(22,27和30℃)和2个相对湿度(40%和55%)组合对DE活性的影响,结果显示30℃和40%相对湿度条件下得到的LT50(110.11 h)最低,说明高温低湿条件下DE活性较高.%Experiments were conducted to enhance the virulence of two isolates of entomopathogenic fungi,IRAN-187C of Beauveria bassiana (Balsamo) Vuillemin, and A-396 isolate of Metarhizium anisopliae (Metschnikow) Sorokin,by combination with InsectoSec(R) formulation of diatomaceous earth (DE) against the adults of Callosobruchus maculatus F.under laboratory conditions.The results demonstrated that the combinations between diatomaceous earth and the fungal isolates at lethal and sub-lethal doses achieved high mortality rates in the adult populations ; moreover,the treatments improved LT50 values of the fungi.The lowest value of LT50 (122.2 h) was achieved by LC50 (B.bassiana,IRAN-187C) + LC50 (DE) treatment which was on par with that of LC50 (M.anisopliae,A-396) + LC50 (DE) combination (128.2 h) based on their 95% fiducial limit values,resulting in better efficacy of each fungal isolate in combination with diatomaceous earth against C.maculatus adults.Also,effects of the three temperatures (22,27,and 30

  8. Virulence of Metarhizium flavoviride 82 to different developmental stages of the brown planthopper, Nilaparvata lugens (Hemiptera:Delphacidae)%黄绿绿僵菌Mf82菌株对不同虫态褐飞虱的毒力

    Institute of Scientific and Technical Information of China (English)

    李茂业; 林华峰; 李世广; 金立

    2012-01-01

    对水稻重要害虫褐飞虱Nilaparvata lugens目前市场上尚无一种理想的微生物杀虫剂.昆虫病原真菌具有从体壁侵入的能力因而对刺吸性害虫的防治具有优势.为此,本研究选用不同原寄主和来源地的3种昆虫病原真菌(金龟子绿僵菌Metarhizium anisopliae、黄绿绿僵菌Metarhizium flavoviride和球孢白僵菌Beauveria bassiana)的12个不同菌株,以1 100孢子/mm2孢子悬浮液进行室内毒力测定.结果表明:在参试的不同菌种12个菌株中,黄绿绿僵菌Mf82菌株对褐飞虱成虫致病力最高,10 d累计校正死亡率为83.5%,致死中时(LT50)为4.6d.其不同浓度孢子液对褐飞虱3个发育阶段有不同程度的致病力,毒力大小顺序为成虫>高龄若虫>低龄若虫.黄绿绿僵菌孢子液对各处理稻株褐飞虱产卵痕部位、卵粒均有侵染作用,10d侵染率分别为66.7%和51.2%,卵龄越低,侵染效果越好,卵龄为0.5d时侵染率最高.本研究表明黄绿绿僵菌Mf82菌株对褐飞虱成虫、若虫和卵均有较强的致病性,是一株极具应用潜力的生防真菌.%At present there still lacks ideal commercial microbial insecticides to control the brown planthopper ( BPH) , Nilaparvata lugens (Stil) , an important pest of rice in Asia. Entomopathogenic fungi can penetrate the cuticle of insect hosts and thus have the advantage of controlling sucking pests. In this study, laboratory bioassays were carried out on 12 isolates of three entomopathogenic fungal species, Metarhizium anisopliae, Metarhizium flavoviride and Beauveria bassiana, which were originally obtained from various hosts and geographic origins, for their virulence against BPH adults. The results showed that at the concentration of 1 100 conidia/mm2, the strain Mf82 had the highest virulence to adults with the corrected mortality reaching 83.5% within 10 d after treatment and the LT50 value 4.65 d. The pathogenicity of Mf82 to adults, nymphs at different instars and

  9. Evaluating the lethal and pre-lethal effects of a range of fungi against adult Anopheles stephensi mosquitoes

    Directory of Open Access Journals (Sweden)

    Blanford Simon

    2012-11-01

    Full Text Available Abstract Background Insecticide resistance is seriously undermining efforts to eliminate malaria. In response, research on alternatives to the use of chemical insecticides against adult mosquito vectors has been increasing. Fungal entomopathogens formulated as biopesticides have received much attention and have shown considerable potential. This research has necessarily focused on relatively few fungal isolates in order to ‘prove concept’. Further, most attention has been paid to examining fungal virulence (lethality and not the other properties of fungal infection that might also contribute to reducing transmission potential. Here, a range of fungal isolates were screened to examine variation in virulence and how this relates to additional pre-lethal reductions in feeding propensity. Methods The Asian malaria vector, Anopheles stephensi was exposed to 17 different isolates of entomopathogenic fungi belonging to species of Beauveria bassiana, Metarhizium anisopliae, Metarhizium acridum and Isaria farinosus. Each isolate was applied to a test substrate at a standard dose rate of 1×109 spores ml-1 and the mosquitoes exposed for six hours. Subsequently the insects were removed to mesh cages where survival was monitored over the next 14 days. During this incubation period the mosquitoes’ propensity to feed was assayed for each isolate by offering a feeding stimulant at the side of the cage and recording the number probing. Results and conclusions Fungal isolates showed a range of virulence to A. stephensi with some causing >80% mortality within 7 days, while others caused little increase in mortality relative to controls over the study period. Similarly, some isolates had a large impact on feeding propensity, causing >50% pre-lethal reductions in feeding rate, whereas other isolates had very little impact. There was clear correlation between fungal virulence and feeding reduction with virulence explaining nearly 70% of the variation in

  10. PROTEOMIC ANALYSIS OF ALLERGENS FROM METARHIZIUM ANISOPLIEA

    Science.gov (United States)

    The goal of this project is the identification and characterization of allergens from the fungus M. Anisopliae, using mass spectrometry (MS). The US EPA, under the "Children at Risk" program, is currently addressing the problem of indoor fungal bioaerosol contamination. One of ...

  11. Preliminary survey for entomopathogenic fungi associated with Ixodes scapularis>/i> (Acari: Ixodidae) in southern New York and New England, USA

    Science.gov (United States)

    Zhioua, Elyes; Ginsberg, Howard S.; Humber, Richard A.; LeBrun, Roger A.

    1999-01-01

    Free-living larval, nymphal, and adult Ixodes scapularis Say were collected from scattered locales in southern New England and New York to determine infection rates with entomopathogenic fungi. Infection rates of larvae, nymphs, males, and females were 0% (571), 0% (272), 0% (57), and 4.3% (47), respectively. Two entomopathogenic fungi were isolated from field-collected I. scapularis females from Fire Island, NY. Isolates were identified as Verticillium lecanii (Zimmermann) Viegas and Verticillium sp. (a member of the Verticillium lecanii species complex).Ixodes scapularis Say is the principal vector of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner (Burgdorfer et al. 1982, Johnson et al. 1984), the etiologic agent of Lyme disease in the northeastern and upper-midwestern United States. Control of I. scapularis is based on chemical treatment (Mather et al. 1987b; Schulze et al. 1987, 1991), environmental management (Wilson et al. 1988, Schulze et al. 1995), and habitat modification (Wilson 1986). These methods have shown variable success, and some potentially have negative environmental effects (Wilson and Deblinger 1993, Ginsberg 1994).Studies concerning natural predators, parasitoids, and pathogens of I. scapularis are rare. The use of ground-dwelling birds as tick predators has had only limited success (Duffy et al. 1992). Nymphal I. scapularis are often infected with the parasitic wasp Ixodiphagus hookeri (Howard) (Mather et al. 1987a, Hu et al. 1993, Stafford et al. 1996, Hu and Hyland 1997), but this wasp does not effectively control I. scapularis populations (Stafford et al. 1996). The entomopathogenic nematodes Steinernema carpocapsae (Weiser) and S. glaseri (Steiner) are pathogenic only to engorged female I. scapularis, and thus have limited applicability (Zhioua et al. 1995). In contrast, the entomogenous fungus Metarhizium anisopliae (Metschnikoff) Sorokin is highly pathogenic to all stages of I. scapularis, unfed as well as engorged

  12. 一株绿僵菌的分离鉴定及对桃小食心虫的致病力研究%Isolation and Identification of a Strain of Metarhizium and Its Virulence to Carposina sasakii

    Institute of Scientific and Technical Information of China (English)

    王刚; 熊琦; 田芬; 谢映平; 薛皎亮

    2013-01-01

    A new strain designated No.TSL06 was isolated and purified from the diseased larvae of the peach fruit moth,Carposina sasakii (Matsumura) (Lepidoptera:Carposinidae),which were collected from a orchard in Shanxi.The strain was identified based on its culture traits,morphological characteristics observed under the optical microscope and scanning electron microscope,and ITS sequence analysis.The identification result showed that the strain belongs to the species of Metarhizium anisopliae (Metsch.) Sorokin.In virulence test,the conidial suspensions in five concentrations were used and the result showed that the maximal correction mortality rate of the infected C.sasakii larvae reached 86.08% in the infected section with concentration of 1.2 × 109 spores/mL.%从山西临汾襄汾景毛乡果园罹病的桃小食心虫Carposina sasakii(Matsumura) (Lepidoptera:Carposinidae)幼虫虫尸中分离纯化获得一株新菌株,命名为TSL06.通过对该菌株的培养性状、光学显微和扫描电镜下的形态特征观察,以及ITS基因序列分析,鉴定出TSL06菌株为绿僵菌属Metarhizium Sorokin的金龟子绿僵菌Metarhizium anisopliae (Metsch.) Sorokin.用该菌株的5种浓度孢子悬液对桃小食心虫幼虫进行毒力试验,结果表明,浓度为1.2×109孢子/mL的孢子悬液对桃小食心虫幼虫的校正致死率最高,达到86.08%.

  13. Fungi with multifunctional lifestyles: endophytic insect pathogenic fungi.

    Science.gov (United States)

    Barelli, Larissa; Moonjely, Soumya; Behie, Scott W; Bidochka, Michael J

    2016-04-01

    This review examines the symbiotic, evolutionary, proteomic and genetic basis for a group of fungi that occupy a specialized niche as insect pathogens as well as endophytes. We focus primarily on species in the genera Metarhizium and Beauveria, traditionally recognized as insect pathogenic fungi but are also found as plant symbionts. Phylogenetic evidence suggests that these fungi are more closely related to grass endophytes and diverged from that lineage ca. 100 MYA. We explore how the dual life cycles of these fungi as insect pathogens and endophytes are coupled. We discuss the evolution of insect pathogenesis while maintaining an endophytic lifestyle and provide examples of genes that may be involved in the transition toward insect pathogenicity. That is, some genes for insect pathogenesis may have been co-opted from genes involved in endophytic colonization. Other genes may be multifunctional and serve in both lifestyle capacities. We suggest that their evolution as insect pathogens allowed them to effectively barter a specialized nitrogen source (i.e. insects) with host plants for photosynthate. These ubiquitous fungi may play an important role as plant growth promoters and have a potential reservoir of secondary metabolites.

  14. Genetic basis of destruxin production in the entomopathogen Metarhizium robertsii.

    Science.gov (United States)

    Giuliano Garisto Donzelli, Bruno; Krasnoff, Stuart B; Moon, Yong-Sun; Sun-Moon, Yong; Churchill, Alice C L; Gibson, Donna M

    2012-04-01

    Destruxins are among the most exhaustively researched secondary metabolites of entomopathogenic fungi, yet definitive evidence for their roles in pathogenicity and virulence has yet to be shown. To establish the genetic bases for the biosynthesis of this family of depsipeptides, we identified a 23,792-bp gene in Metarhizium robertsii ARSEF 2575 containing six complete nonribosomal peptide synthetase modules, with an N-methyltransferase domain in each of the last two modules. This domain arrangement is consistent with the positioning of the adjacent amino acids N-methyl-L: -valine and N-methyl-L: -alanine within the depsipeptide structure of destruxin. DXS expression levels in vitro and in vivo exhibited comparable patterns, beginning at low levels during the early growth phases and increasing with time. Targeted gene knockout using Agrobacterium-mediated transformation produced mutants that failed to synthesize destruxins, in comparison with wild type and ectopic control strains, indicating the involvement of this gene in destruxin biosynthesis. The destruxin synthetase (DXS) disruption mutant was as virulent as the control strain when conidial inoculum was topically applied to larvae of Spodoptera exigua, Galleria mellonella, and Tenebrio molitor indicating that destruxins are dispensable for virulence in these insect hosts. The DXS mutants exhibited no other detectable changes in morphology and development.

  15. Genome Studies on Nematophagous and Entomogenous Fungi in China

    Directory of Open Access Journals (Sweden)

    Weiwei Zhang

    2016-02-01

    Full Text Available The nematophagous and entomogenous fungi are natural enemies of nematodes and insects and have been utilized by humans to control agricultural and forestry pests. Some of these fungi have been or are being developed as biological control agents in China and worldwide. Several important nematophagous and entomogenous fungi, including nematode-trapping fungi (Arthrobotrys oligospora and Drechslerella stenobrocha, nematode endoparasite (Hirsutella minnesotensis, insect pathogens (Beauveria bassiana and Metarhizium spp. and Chinese medicinal fungi (Ophiocordyceps sinensis and Cordyceps militaris, have been genome sequenced and extensively analyzed in China. The biology, evolution, and pharmaceutical application of these fungi and their interacting with host nematodes and insects revealed by genomes, comparing genomes coupled with transcriptomes are summarized and reviewed in this paper.