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Sample records for bassiana isolates obtained

  1. Antimicrobial activity of Stomoxys calcitransagainst Beauveria bassiana sensu lato isolates

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    Ana Paula Rodrigues de Moraes

    Full Text Available This study had the aims of evaluating the antimicrobial characteristics of Stomoxys calcitrans (Diptera: Muscidae larvae against the fungal isolates CG138, CG228 and ESALQ986 of Beauveria bassiana sensu lato (Balsamo-Crivelli Vuillemin, 1912 (Hypocreales: Cordycipitaceae. S. calcitrans eggs, larvae and pupae were exposed to these same isolates. Statistical analysis showed that the immature stages of S. calcitrans were not susceptible to the fungal isolates used, regardless of the exposure method. Diffusion test on solid culture medium reveled that macerated S. calcitrans larvae exposed to isolate CG138 reduced CG138 fungal development. The analysis of the chromatographic profiles indicated that the macerate or mucus of larvae of the control group and the groups exposed to the isolate CG138 presented different profiles. Reduced development of the isolate CG138 on the larvae cuticle was observed by means of scanning electron microscopy.

  2. Genetic diversity among brazilian isolates of beauveria bassiana: comparisons with non-brazilian isolates and other beauveria species

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    Fernandes, E.K.K.; Moraes, A.M.L.; Pacheco, R.S.; Rangel, D.E.N.; Miller, M.P.; Bittencourt, V.R.E.P.; Roberts, D.W.

    2009-01-01

    Aims: The genetic diversity of Beauveria bassiana was investigated by comparing isolates of this species to each other (49 from different geographical regions of Brazil and 4 from USA) and to other Beauveria spp. Methods and Results: The isolates were examined by multilocus enzyme electrophoresis (MLEE), amplified fragment length polymorphism (AFLP), and rDNA sequencing. MLEE and AFLP revealed considerable genetic variability among B. bassiana isolates. Several isolates from South and Southeast Brazil had high similarity coefficients, providing evidence of at least one population with clonal structure. There were clear genomic differences between most Brazilian and USA B. bassiana isolates. A Mantel test using data generated by AFLP provided evidence that greater geographical distances were associated with higher genetic distances. AFLP and rDNA sequencing demonstrated notable genotypic variation between B. bassiana and other Beauveria spp. Conclusion: Geographical distance between populations apparently is an important factor influencing genotypic variability among B. bassiana populations in Brazil. Significance and Impact of the Study: This study characterized many B. bassiana isolates. The results indicate that certain Brazilian isolates are considerably different from others and possibly should be regarded as separate species from B. bassiana sensu latu. The information on genetic variation among the Brazilian isolates, therefore, will be important to comprehending the population structure of B. bassiana in Brazil. ?? 2009 The Society for Applied Microbiology.

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

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    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.

  4. Characterization of Beauveria bassiana (Ascomycota: Hypocreales) isolates associated with Agrilus planipennis (Coleoptera: Buprestidae) populations in Michigan

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    Louela A. Castrillo; Leah S. Bauer; Houping Liu; Michael H. Griggs; John D. Vandenberg

    2010-01-01

    Earlier research in Michigan on fungal entomopathogens of the emerald ash borer (EAB), a major invasive pest of ash trees, resulted in the isolation of Beauveria bassiana from late-instar larvae and pre-pupae. In the present study, some of these isolates were characterized and compared to ash bark- and soil-derived isolates to determine their...

  5. Polymorphism of Beauveria bassiana (Deuteromycota: Hyphomycetes) strains isolated from Ixodes ricinus (Acari: Ixodidae) in Moldova.

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    Mitina, G V; Tokarev, Y S; Movila, A A; Yli-Mattila, T

    2011-03-01

    Polymorphism of 10 Beauveria bassiana strains, isolated from Ixodes ricinus in Moldova, was evaluated using traditional (morphological and cultural properties) and molecular (RAPD patterns and ITS sequences) methods. The isolates differed greatly in morphological and cultural features, such as color, consistence, and growth rate. Four RAPD-PCR markers were used to evaluate genetic diversity of the strains. Phylogenetic neighbor-joining analysis of RAPD patterns divided strains into 3 major clades. The ITS sequences of 8 strains were identical to those of known B. bassiana strains. Two subsets (1 and 2) different by one nucleotide change were found in the ITS1 region. One strain of subset 1 was different from known B. bassiana strains by possessing 2 point mutations in the ITS region. RAPD-based clustering correlated to ITS sequence and colony morphology-based grouping of the strains.

  6. Evaluation of Beauveria bassiana (Hyphomycetes) isolates as potential agents for control of Dendroctonus valens

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    Long-Wa Zhang; Yu-Jun Liu; Jian Yao; Bin Wang; Bo Huang; Zeng-Zhi Li; Mei-Zhen Fan; Jiang-Hua Sun

    2011-01-01

    The red turpentine beetle (RTB), Dendroctonus valens LeConte, as a destructive invasive pest, has become one of the most economically important forest pest in China. Effective control measures are desperately needed. Entomopathogenic fungi, such as Beauveria bassiana, have shown great potential for the management of some bark beetle species. In this study, 12 isolates of B. bassiana from bark beetle were examined for biological characteristics and virulence, to assess their potential as biocontrol agents for RTB. There were significant differences (at P = 0.05) in colony growth rate, conidial yield, conidial germination, tolerance to UV light and extracellular proteases activity among the tested B. bassiana isolates. Isolates, including Bbl801, Bbl906, Bb789 and Bb773, exhibited the best characteristics, because they have faster hyphal growth rate, higher spore production and faster spore germination, higher UV tolerance and protease (Prl) production. The results of a pathogenicity test of B. bassiana on RTB larvae showed that most isolates of B. bassiana have demonstrated high efficacy and the highest virulent isolate was Bbl801, which killed 100% of the treated insects and had a median lethal time (LT50) of 4.60 days at a concentration of 1× 107 conidia/mL. Therefore, isolate Bb 1801 has a great potential for sustainable control of RTB in the forest. The correlation between biological characteristics and virulence of the fungal isolates is discussed and the possibility of combination of entomopathogenic fungi with semiochemicals, as one of the promising strategy for RTB control, is considered.

  7. Endophytic Ability of Different Isolates of Entomopathogenic Fungi Beauveria bassiana (Balsamo) Vuillemin in Stem and Leaf Tissues of Maize (Zea mays L.).

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    Renuka, S; Ramanujam, B; Poornesha, B

    2016-06-01

    The present study was conducted to examine the ability of six promising indigenous isolates of Beauveria bassiana (NBAII-Bb-5a, 7, 14, 19, 23 and 45) as an endophyte in maize stem and leaf tissues. Maize seedlings (var. Nithyashree) were inoculated with conidial suspensions and were examined for endophytic establishment in leaf and stems at different intervals during 15-90 days after treatment. All six isolates showed colonization in stem and leaf tissues with varying abilities of colonization and persistence. The mean percent colonization ranged from 7.41 to 20.37 % in older stem tissues and 3.70 to 21.29 % in young stem tissues and in leaf, it ranged from 6.46 to 27.78 % in older leaf tissues and 11.11 to 26.85 % in young leaf tissues. Among six isolates tested, Bb-23 isolate recorded the maximum mean colonization in older stem (20.37 %), older leaf (27.78 %) and in young stem (21.29 %). Bb-5a isolate showed maximum mean colonization in young leaf tissues (26.85 %). Persistence of inoculated fungal isolates decreased with increase in age of the plant. No physical symptoms of damage were observed in any of the B. bassiana treated plants. No colonization of B. bassiana was observed in the untreated control maize plants. The results obtained in plating and PCR techniques were similar with regard to the confirmation of endophytic establishment of B. bassiana. This study indicated the possibility of using B. bassiana as an endophyte in maize for management of maize stem borer, Chilo partellus.

  8. Isolation and characterization of 2-pyridone alkaloids and alloxazines from Beauveria bassiana.

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    Andrioli, W J; Lopes, A A; Cavalcanti, B C; Pessoa, C; Nanayakkara, N P D; Bastos, J K

    2016-12-29

    Two novel compounds bearing heterocyclic nitrogen, 2-pyridone alkaloid (1) and alloxazine derivative (2), along with the known pretenellin B (3), pyridovericin (4) and lumichrome (5) were isolated from a culture of the entomopathogenic fungal strain Beauveria bassiana. The chemical structures of 2-pyridone alkaloid and alloxazine derivative were established on the basis of the interpretation of spectroscopic data. The isolated compounds were evaluated in a panel of five cancer cell lines and pyridovericin exhibited cytotoxicity (IC50, μM) against cancer cell lines: HL-60 (25.9 ± 0.3), HCT8 (34.6 ± 3.6), MDA-MB435 (34.8 ± 3.8) and SF295 (31.1 ± 0.6). Considering that other pyridone compounds display good cytotoxic activity, it would be suggested to obtain new semi synthetic derivatives of pyridovericin, for the development of new cytotoxic chemical entities.

  9. Comparative genomics of Beauveria bassiana

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    Valero-Jiménez, Claudio A.; Faino, Luigi; Spring in 'T Veld, Daphne; Smit, Sandra; Zwaan, Bas J.; Kan, van Jan A.L.

    2016-01-01

    Background: Entomopathogenic fungi such as Beauveria bassiana are promising biological agents for control of malaria mosquitoes. Indeed, infection with B. bassiana reduces the lifespan of mosquitoes in the laboratory and in the field. Natural isolates of B. bassiana show up to 10-fold differences

  10. Comparative genomics of Beauveria bassiana

    NARCIS (Netherlands)

    Valero-Jiménez, Claudio A.; Faino, Luigi; Spring in 'T Veld, Daphne; Smit, Sandra; Zwaan, Bas J.; Kan, van Jan A.L.

    2016-01-01

    Background: Entomopathogenic fungi such as Beauveria bassiana are promising biological agents for control of malaria mosquitoes. Indeed, infection with B. bassiana reduces the lifespan of mosquitoes in the laboratory and in the field. Natural isolates of B. bassiana show up to 10-fold differences

  11. Responses to abiotic environmental stresses among phylloplane and soil isolates of Beauveria bassiana from two holm oak ecosystems.

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    Fernández-Bravo, María; Garrido-Jurado, Inmaculada; Valverde-García, Pablo; Enkerli, Jürg; Quesada-Moraga, Enrique

    2016-11-01

    The response of entomopathogenic mitosporic ascomycete (EMAs) to abiotic stresses might be adapted to the microhabitats in which they inhabit. In phylloplane, these organisms are more exposed to such stresses than they are in soil, which may have led to adaptation to this environment. In the present work, we investigate whether Beauveria bassiana genotype or isolation habitat, i.e., soil or phylloplane, within the same geographic area influences their responses to key environmental stresses, such as temperature, moisture and ultraviolet radiation (UV-B), which can affect their successful use in microbial control. Twenty isolates of B. bassiana obtained from the soil and phylloplane in two ecosystems from southern Spain (holm oak dehesa and a reforested area) were selected to study the population distribution of these isolates and evaluate their thermal, humidity and UV-B requirements. Molecular characterization was conducted by using elongation factor-1α (EF-1α), the intergenic nuclear region Bloc and 15 microsatellite primers. The cluster analysis based on concatenated EF-1α and Bloc sequences grouped the 20 isolates into five clades within B. basiana, with Clades a, b, d and e containing both soil and phylloplane isolates and Clade c including three phylloplane isolates. The dendrogram and the minimal spanning network generated from the genetic distances among multilocus genotypes showed four divergent groups corresponding to the five clades obtained based on the sequence data (Clades b and d were represented in the same group), with a high degree of shared alleles within groups and few alleles shared among groups. Although no relationship was found between MLG and the habitat (soil or phylloplane) of isolation, isolates grouped into Clade c, all of which were collected from phylloplane, formed a separate group of MLGs. To investigate our hypothesis, the responses to temperature (germination and colony growth evaluated in the range 15-35°C), water activity

  12. Label-Free Differential Proteomics and Quantification of Exoenzymes from Isolates of the Entomopathogenic Fungus Beauveria bassiana

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    Dionisio, Giuseppe; Kryger, Per; Steenberg, Tove

    2016-01-01

    Beauveria bassiana is an entomopathogenic fungus that grows both in vivo and in vitro. In vivo it can colonize live insect hosts, and tissue digestion occurs by secreted hydrolytic exoenzymes. It can also colonize dead insect tissue provided this is free from competing microorganisms. Depending on whether the host is alive or dead the expression (quality/quantity) of the exoenzymes may vary. We have grown several isolates of B. bassiana in shaking flasks for 120 h at 25 °C in order to evaluate the maximal exoenzyme production using two diet regimes. As sole carbon, nitrogen, and phosphate sources we used 1% shrimp chitin and either 0.5% w/v of dead intact American cockroach (Periplaneta americana) or their isolated cuticles. This is the first report of a differential proteomics of B. bassiana exoenzymes performed by label-free nano-LC MS/MS. Total proteolytic enzyme activity was mainly due to Pr1A or Pr1B depending on the isolate and the diet regime. The most differentially secreted enzymes were: the cuticle-degrading subtilisin Pr1A, GH13 alpha-glycosidase, glucan endo-1,3-beta-glucosidase, subtilisin-like proteinase Spm1, lipase 1, beta-1,3 exoglucanase, and endo-1,3-beta-glucosidase. Among the B. bassiana isolates analyzed, Bb 678 and Bb BG were the most active in Pr1A secretion. PMID:27754403

  13. Label-Free Differential Proteomics and Quantification of Exoenzymes from Isolates of the Entomopathogenic Fungus Beauveria bassiana.

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    Dionisio, Giuseppe; Kryger, Per; Steenberg, Tove

    2016-10-14

    Beauveria bassiana is an entomopathogenic fungus that grows both in vivo and in vitro. In vivo it can colonize live insect hosts, and tissue digestion occurs by secreted hydrolytic exoenzymes. It can also colonize dead insect tissue provided this is free from competing microorganisms. Depending on whether the host is alive or dead the expression (quality/quantity) of the exoenzymes may vary. We have grown several isolates of B. bassiana in shaking flasks for 120 h at 25 °C in order to evaluate the maximal exoenzyme production using two diet regimes. As sole carbon, nitrogen, and phosphate sources we used 1% shrimp chitin and either 0.5% w/v of dead intact American cockroach (Periplaneta americana) or their isolated cuticles. This is the first report of a differential proteomics of B. bassiana exoenzymes performed by label-free nano-LC MS/MS. Total proteolytic enzyme activity was mainly due to Pr1A or Pr1B depending on the isolate and the diet regime. The most differentially secreted enzymes were: the cuticle-degrading subtilisin Pr1A, GH13 alpha-glycosidase, glucan endo-1,3-beta-glucosidase, subtilisin-like proteinase Spm1, lipase 1, beta-1,3 exoglucanase, and endo-1,3-beta-glucosidase. Among the B. bassiana isolates analyzed, Bb 678 and Bb BG were the most active in Pr1A secretion.

  14. Effect of temperature and humidity on pathogenicity of native Beauveria bassiana isolate against Musca domestica L.

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    Mishra, Sapna; Kumar, Peeyush; Malik, Anushree

    2015-12-01

    Beauveria bassiana HQ917687 virulence to housefly larvae and adult was assessed at different relative humidity, RH (50, 75, 90, and 100 %) and temperature (15, 20, 25, 30, 35, 40, 45 °C) conditions at the fungal dose of 10(8) conidia/ml. Depending on the temperature and RH regime tested, difference in mortality rates of housefly adult and larvae were detected. During assay on adult housefly, 100 % mortality was achieved at RH, 90 and 100 % while the temperature of 30 °C showed maximum mortality at all the tested humidity conditions. Lethal time, LT50 was 2.9 days at 100 % RH. Larval mortality at different humidity conditions varied between 30 and 74 %, with maximum mortality at 100 % RH and 30 °C. Optimum temperature for B. bassiana virulence to housefly larvae was also found to be 30 °C. The interaction between temperature and RH revealed significant effect of RH at moderate temperature range (20-35 °C), while such an interaction was not observed at extreme temperatures. The results obtained in this study have useful implications in understanding the pathogen behavior under actual field conditions. This in turn may help devising suitable entomopathogen release schedules for maximum fungal infection.

  15. Successful Development of Cordyceps bassiana Stromata from Beauveria bassiana

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    Lee, Je-O; Shrestha, Bhushan; Sung, Gi-Ho; Han, Sang-Kuk

    2010-01-01

    A specimen of Beauveria bassiana was collected from Yang-yang of Gangwon province, Korea in October 2006. Conidial isolates were prepared from the specimen by the dilution method and inoculated in brown rice medium for fruiting body production. After nearly two months incubation for perithecial stromata developed from single isolates as well as from their combinations. They were determined as Cordyceps bassiana by observing the stromatal characters and their conidial structures. This is the first report of the development of C. bassiana from B. bassiana cultures. PMID:23956619

  16. Nutritional optimization of a native Beauveria bassiana isolate (HQ917687) pathogenic to housefly, Musca domestica L.

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    Mishra, Sapna; Malik, Anushree

    2013-10-01

    A new Beauveria bassiana isolate, showing high activity against Musca domestica L. (Diptera: Muscidae) adults (mortality-100.0 %), larvae (mortality-72.3 %) and pupae (Infection in emerged flies-96.7 %) was used. The isolate was subjected to a combinational approach towards selection of process parameters for its growth optimization. Initial screening of several carbon and nitrogen sources revealed glucose and NaNO3 as the most suitable source for optimal biomass and spore production. Further, optimization through Placket-Burman and a 2(5) full factorial central composite design revealed highly significant effect of glucose and pH. The optimum composition for maximum biomass yield was (g/l): glucose 28; NaNO3 2.43; KH2PO4 1.32; MgSO4 0.60; and pH 7.00. Glucose concentration showed almost linear relationship with biomass yield, indicating its significant contribution in medium composition for fungal growth. Highly significant interactions were observed between glucose and pH, followed by glucose and NaNO3 concentration.

  17. Isolation and Assessment of Stability of Six Formulations of Entomopathogenic Beauveria bassiana.

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    Mwamburi, Lizzy A

    2016-01-01

    Beauveria bassiana is the most widely studied and exploited entomopathogen. The development of a suitable formulation for B. bassiana is a critical component in aiding the entomopathogen germinate and infect the host. In addition to being economical to produce, having high residual activity, it is also important that the formulation is easy to handle, stable during storage, and convenient to mix and apply and be consistently effective in controlling the target pest.In this chapter we describe preparation of experimental formulations of conidia of B. bassiana. The formulations are prepared with barley, rice, wheat bran, clay, kaolin, and peat. The protocol for assessing the stability of the formulations of B. bassiana is also described.

  18. Beauveria bassiana: Quercetinase production and genetic diversity

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    Eula Maria de M. B., Costa; Fabiana Cristina, Pimenta; Christian, Luz; Valéria de, Oliveira; Marília, Oliveira; Elda, Bueno; Silvana, Petrofeza

    2011-01-01

    Beauveria bassiana genetic diversity and ability to synthesize quercetin 2,3-dioxygenase (quercetinase) were analyzed. B. bassiana isolates, obtained from Brazilian soil samples, produced quercetinase after induction using 0.5 g/L quercetin. B. bassiana ATCC 7159 (29.6 nmol/mL/min) and isolate IP 11 (27.5 nmol/ml/min) showed the best performances and IP 3a (9.5 nmol/mL/min) presented the lowest level of quercetinase activity in the culture supernatant. A high level of polymorphism was detected by random amplified polymorphic DNA (RAPD) analysis. The use of internal-transcribed-spacer ribosomal region restriction fragment length polymorphism (ITS-RFLP) did not reveal characteristic markers to differentiate isolates. However, the ITS1-5.8S-ITS2 region sequence analysis provided more information on polymorphism among the isolates, allowing them to be clustered by relative similarity into three large groups. Correlation was tested according to the Person's correlation. Data of our studies showed, that lower associations among groups, level of quercetinase production, or geographical origin could be observed. This study presents the production of a novel biocatalyst by B. bassiana and suggests the possible industrial application of this fungal species in large-scale biotechnological manufacture of quercetinase. PMID:24031599

  19. Beauveria bassiana: quercetinase production and genetic diversity

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    Eula Maria de M. B Costa

    2011-03-01

    Full Text Available Beauveria bassiana genetic diversity and ability to synthesize quercetin 2,3-dioxygenase (quercetinase were analyzed. B. bassiana isolates, obtained from Brazilian soil samples, produced quercetinase after induction using 0.5 g/L quercetin. B. bassiana ATCC 7159 (29.6 nmol/mL/min and isolate IP 11 (27.5 nmol/ml/min showed the best performances and IP 3a (9.5 nmol/mL/min presented the lowest level of quercetinase activity in the culture supernatant. A high level of polymorphism was detected by random amplified polymorphic DNA (RAPD analysis. The use of internal-transcribed-spacer ribosomal region restriction fragment length polymorphism (ITS-RFLP did not reveal characteristic markers to differentiate isolates. However, the ITS1-5.8S-ITS2 region sequence analysis provided more information on polymorphism among the isolates, allowing them to be clustered by relative similarity into three large groups. Correlation was tested according to the Person's correlation. Data of our studies showed, that lower associations among groups, level of quercetinase production, or geographical origin could be observed. This study presents the production of a novel biocatalyst by B. bassiana and suggests the possible industrial application of this fungal species in large-scale biotechnological manufacture of quercetinase.

  20. 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)

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    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)

  1. Pathogenicity of Beauveria bassiana isolated from Moroccan Argan forests soil against larvae of Ceratitis capitata (Diptera: Tephritidae) in laboratory conditions.

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    Imoulan, Abdessamad; Elmeziane, Abdellatif

    2014-03-01

    The Mediterranean fruit fly, Ceratitis capitata Wiedemann (Diptera: Tephritidae), is the major tephritid pest in Morocco. This pest survives in Moroccan forests Argania spinosa and continually invades the nearest agricultural areas. Entomopathogenic fungi are an interesting tool for fruit fly control and hold a useful alternative to conventional insecticides. However, primary selection of effective pathogens should be taken in laboratory condition prior to applying them in the field. Here, we used third late instar larvae of C. capitata to investigate the effectiveness of 15 local Beauveria bassiana isolates. Results showed that all isolates were able to infect the larval stage, producing a large mortality rate in puparia ranging from 65 to 95 % and caused significant reduction in adult emergence. The fungal treatments revealed that the mycosis occurred also in adults escaping infection as pupariating larvae. The percentage of mycosed puparia was highest in strain TAM6.2 (95 %) followed by ERS4.16 (90 %), therefore they were the most virulent. Median lethal concentration (LC₅₀) was studied for five isolates at four concentrations ranging from 10⁵ to 10⁸ conidia ml⁻¹. The results showed that the slopes of regression lines for B. bassiana ERS4.16 (slope = 0.386) and TAM6.2 (slope = 0.41) were the most important and had the lowest LC₅₀ values (2.85 × 10³ and 3.16 × 10³ conidia ml⁻¹ respectively). This investigation suggests that the soil of Argan forests contains pathogenic B. bassiana isolates and highlights for the first time their potential as biological control toward C. capitata larval stage in Morocco.

  2. Estimation of median lethal concentration of three isolates of Beauveria bassiana for control of Megacopta cribraria (Heteroptera: Plataspidae) bioassayed on solid Lugys Spp. Diet²

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    The entomopathogenic fungus Beauveria bassiana is a potential candidate for biological control of a variety of pests. The objectives of this study were isolate and identify the fungus from Kudzu bugs and determine its infectivity comparing with two highly pathogenic isolates including the Mississipp...

  3. Estimation of Median Lethal Concentration of Three Isolates of Beauveria bassiana for Control of Megacopta cribraria (Heteroptera: Plataspidae Bioassayed on Solid Lygus spp. Diet

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    Maribel Portilla

    2016-06-01

    Full Text Available The kudzu bug, Megacopta cribraria (F., is an urban nuisance and significant agricultural pest. The median lethal concentrations of three strains of Beauveria bassiana (Balsamo, including the Mississippi Delta native strain (NI8 isolated from Lygus lineolaris (Palisot de Beauvois, the commercial strain BotaniGard® (GHA (Victor, NY, USA, and the B. bassiana strain isolated from M. cribraria (KUDSC, were estimated on kudzu bug adults. A technique developed to evaluate B. bassiana against L. lineolaris was used. Younger adults (eight days after collection were treated with NI8 and GHA and older adult (50 days after collection were treated with NI8, GHA and KUDSC. Higher concentrations (n × 106, n × 107 of NI8 and GHA caused kudzu bug mortality two days after treatment in younger adults and similar concentrations of NI8, GHA, and KUDSC caused mortality one day after treatment in older adults. Lower concentrations (n × 104, n × 105 were not significantly different in mortality between strains. LS50 values of the KUDSC were significantly lower than NI8 and GHA values in older adults. This is the first available information on median lethal concentration of B. bassiana on kudzu bug adults bioassayed on artificial diet. It was determined that B. bassiana (KUDSC and NI8 are highly effective for young adults at very low doses (LC50 1.98–4.98 viable spores per mm2.

  4. Estimation of Median Lethal Concentration of Three Isolates of Beauveria bassiana for Control of Megacopta cribraria (Heteroptera: Plataspidae) Bioassayed on Solid Lygus spp. Diet.

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    Portilla, Maribel; Jones, Walker; Perera, Omaththage; Seiter, Nick; Greene, Jeremy; Luttrell, Randall

    2016-06-30

    The kudzu bug, Megacopta cribraria (F.), is an urban nuisance and significant agricultural pest. The median lethal concentrations of three strains of Beauveria bassiana (Balsamo), including the Mississippi Delta native strain (NI8) isolated from Lygus lineolaris (Palisot de Beauvois), the commercial strain BotaniGard(®) (GHA) (Victor, NY, USA), and the B. bassiana strain isolated from M. cribraria (KUDSC), were estimated on kudzu bug adults. A technique developed to evaluate B. bassiana against L. lineolaris was used. Younger adults (eight days after collection) were treated with NI8 and GHA and older adult (50 days after collection) were treated with NI8, GHA and KUDSC. Higher concentrations (n × 10⁶, n × 10⁷) of NI8 and GHA caused kudzu bug mortality two days after treatment in younger adults and similar concentrations of NI8, GHA, and KUDSC caused mortality one day after treatment in older adults. Lower concentrations (n × 10⁴, n × 10⁵) were not significantly different in mortality between strains. LS50 values of the KUDSC were significantly lower than NI8 and GHA values in older adults. This is the first available information on median lethal concentration of B. bassiana on kudzu bug adults bioassayed on artificial diet. It was determined that B. bassiana (KUDSC and NI8) are highly effective for young adults at very low doses (LC50 1.98-4.98 viable spores per mm²).

  5. Stable Formation of Fruiting Body in Cordyceps bassiana

    Science.gov (United States)

    Lee, Je-O; Shrestha, Bhushan; Kim, Tae-Woong; Sung, Gi-Ho

    2007-01-01

    In order to breed a Cordyceps bassiana isolate that stably forms fruiting body in artificial cultivation, isolates derived from subculturing and single spores were tested through mating. From C. bassiana EFCC 783, three subcultured isolates EFCC 2830, EFCC 2831 and EFCC 2832 were obtained and fourteen single conidial isolates were obtained from these three subcultured isolates. Two different morphological types were found in the fourteen single conidial isolates. One type was able to form synnemata and another type was not able to form synnemata. Since switch of morphological type was not observed despite their continuous subculturing, cross was performed between the two types and the formation of fruiting body was examined. Ascospores were obtained from a selected fruiting body formed by hybrid of the cross. Self-cross and combinational cross of the ascospore-derived isolates generated hybrids that stably produce high quality fruiting body in artificial media. PMID:24015103

  6. Temperature and moisture requirements for conidial germination of an isolate of Beauveria bassiana, pathogenic to Rhodnius prolixus.

    Science.gov (United States)

    Luz, C; Fargues, J

    1997-01-01

    The effects of temperature, relative humidity and water activity on germination of conidia of an isolate of Beauveria bassiana (Bals.) Vuill. pathogenic to the triatomine vector of Chagas' disease, Rhodnius prolixus Stål., were investigated in vitro. Germination occurred at temperatures between 15 degrees C and 35 degrees C under saturated atmosphere and the optima ranged from 25 degrees C to 30 degrees C. At the extreme temperatures tested (15 degrees C and 35 degrees C) the germination process was delayed, but germination rates reached more than 95%. Germination of B. bassiana conidia was strongly affected by moisture conditions. The availability of water, in both atmospheric and liquid conditions, caused changes in germination times as well as in germination rates. For example, at 25 degrees C + 0.5 degrees C, germination took place within 20 h at 95.5% RH, whereas it needed 72 h of incubation at 90% RH. Germination times increased as the water activity declined from 0.96 a(w) to 0.92 a(w). Below 0.92 a(w), no germination was observed after a 72 h incubation time.

  7. 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

  8. 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.

  9. Pro-Apoptotic Activity of 4-Isopropyl-2-(1-Phenylethyl) Aniline Isolated from Cordyceps bassiana.

    Science.gov (United States)

    Kim, Mi Seon; Lee, Yunmi; Sung, Gi-Ho; Kim, Ji Hye; Park, Jae Gwang; Kim, Han Gyung; Baek, Kwang Soo; Cho, Jae Han; Han, Jaegu; Lee, Kang-Hyo; Hong, Sungyoul; Kim, Jong-Hoon; Cho, Jae Youl

    2015-07-01

    Cordyceps species including Cordyceps bassiana are a notable anti-cancer dietary supplement. Previously, we identified several compounds with anti-cancer activity from the butanol fraction (Cb-BF) of Cordyceps bassiana. To expand the structural value of Cb-BF-derived anti-cancer drugs, we employed various chemical moieties to produce a novel Cb-BF-derived chemical derivative, KTH-13-amine-monophenyl [4-isopropyl-2-(1-phenylethyl) aniline (KTH-13-AMP)], which we tested for anti-cancer activity. KTH-13-AMP suppressed the proliferation of MDA-MB-231, HeLa, and C6 glioma cells. KTH-13-AMP also dose-dependently induced morphological changes in C6 glioma cells and time-dependently increased the level of early apoptotic cells stained with annexin V-FITC. Furthermore, the levels of the active full-length forms of caspase-3 and caspase-9 were increased. In contrast, the levels of total forms of caspases-3, caspase-8, caspase-9, and Bcl-2 were decreased in KTH-13-AMP treated-cells. We also confirmed that the phosphorylation of STAT3, Src, and PI3K/p85, which is linked to cell survival, was diminished by treatment with KTH-13-AMP. Therefore, these results strongly suggest that this compound can be used to guide the development of an anti-cancer drug or serve as a lead compound in forming another strong anti-proliferative agent.

  10. Rosalia longicorn Rosalia alpina (LINNAEUS, 1758 (Coleoptera: Cerambycidae as a host of the entomopathogenic fungus Cordyceps bassiana LI, LI, HUANG

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    Bartnik Czesław

    2016-06-01

    Full Text Available The paper describes, for the first time, the occurrence of the entomopathogenic fungus Cordyceps bassiana (anamorph: Beauveria bassiana on the imago of the endangered beetle Rosalia longicorn Rosalia alpina from the Low Beskid Mountains (the Carpathians, SE Poland. Furthermore, an isolate of the saprotrophic fungus Hypoxylon fragiforme was obtained as a result of laboratory tests on R. alpina specimens. Relationships between the identified fungi and R. alpina are discussed.

  11. 不同宿主的白僵菌菌株分离及特性研究%A study on Beauveria bassiana strains isolated in different hosts and their biological characteristics//

    Institute of Scientific and Technical Information of China (English)

    查玉平; 陈京元; 蔡三山; 赵东容

    2011-01-01

    9 strains of Beaiweria bassiana isolated in different hosts and their biological characteristics were studied. The results showed that there were 8 strains of B. bassiana ( Bals. ) Vuillemin, and a strain of B. brongniartii (Sacc. ) Petch. Based on B. bassiana colony characteristics, vegetative growth and sporulation, the 2nd and 3rd strains were fit for the production.%以9株白僵菌菌株为材料,对其进行分离鉴定并探讨菌株的生物学特性.结果表明:有1株为布氏白僵菌(Beauveria brongniartii),8株为球孢白僵菌(B.bassiana).结合白僵菌菌落特征、营养生长情况和产孢量来看,2号和3号菌株比较适合应用于生产.

  12. Selection, assessment of virulence to Alphitobius diaperinus, and Pr1 enzyme production of Beauveria bassiana (Bals. Vuill. isolates cultured at stress temperatures

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    Kelly Christiane Constanski

    2015-12-01

    Full Text Available The entomopathogenic fungus Beauveria bassiana is a promising agent for use in insect control. Its pathogenic activity, as well as other factors such as temperature that can interfere with its development, should be assessed, thus, establishing the foundations for B. bassiana use in biological control programs. The objective of this study was to select and induce tolerance of B. bassiana isolates to high and low temperatures and to assess their virulence before and after exposure to those temperatures. A pre-selection test was performed, in which the tolerance of isolates to stress temperatures was tested and compared to the ideal growth temperature of 25 °C for this organism. For the isolates/temperature combinations resulting in growth, conidia germination and colony-forming units (CFUs were assessed. The isolates Unioeste 4 and Unioeste 40 exhibited >95% germinated conidia at 16 and 31 °C. Thereafter, they underwent four consecutive passages at maximum and minimum tolerated temperatures (10 and 37 °C. A significant difference in germination was observed between the two isolates at all temperatures tested. More CFUs were observed for Unioeste 4 compared to Unioeste 40 at all temperatures, and in the case of the latter, there was no difference in CFU formation at 10 and 25 °C. For both isolates, decreased vegetative growth was observed at 37 °C. Recovery of virulence was observed in both isolates, as determined by insect mortality. No relationship was observed between production of the enzyme Pr1 and the virulence of the isolates.

  13. 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.

  14. Patogenicidade de isolados de Beauveria bassiana para ovos, larvas e ninfas ingurgitadas de Rhipicephalus sanguineus Pathogenicity of Beauveria bassiana isolates towards eggs, larvae and engorged nymphs of Rhipicephalus sanguineus

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    Nancy Prette

    2005-08-01

    Full Text Available O presente trabalho objetivou avaliar a ação dos isolados JAB 07, CB 7 e AM 9 do fungo Beauveria bassiana para ovos, larvas e ninfas ingurgitadas de Rhipicephalus sanguineus. Padronizou-se a concentração de 109 conídios mL-1 para cada isolado e por meio de diluições seriadas, obtiveram-se as suspensões com concentrações de 10(8 e 10(7con. mL-1. Preparou-se um bioensaio para cada fase do ciclo de vida do carrapato. Cada ensaio foi composto por 10 grupos tratamentos, formados por um isolado do fungo e uma concentração de conídios, e um grupo controle contendo apenas o veículo das suspensões. Os isolados do fungo, aplicados nas diferentes concentrações de esporos, causaram redução no percentual de eclosão de larvas oriundas de ovos infectados (0,7 a 12,1% de eclosão e no percentual de ecdise de larvas (4,7 a 33,7% de ecdise e ninfas (0 a 16,7% de ecdise. Não houve diferença entre os isolados (P>0,05 quanto à infecção de ovos, mas a aplicação de 109 con./mL de JAB 07 e AM 9 promoveu redução significativa da eclosão de larvas, em relação à concentração de 10(7con. mL-1. O isolado CB 7 foi o mais eficaz na redução do número de ecdises de larvas. Os isolados e as concentrações de conídios não diferiram (P>0,05 quanto à capacidade de redução do percentual de ecdise de ninfas, mas evidenciou-se intensa atividade patogênica do fungo.The aim of the present study was to determine the action of isolates JAB 07, CB 7 and AM 9 of the fungus Beauveria bassiana on eggs, larvae and engorged nymphs of Rhipicephalus sanguineus tick. A concentration of 109 conidia mL-1 was standardized for each isolate and suspensions containing 10(8 and 10(7 conidia mL-1 were obtained by serial dilution. A bioassay was prepared for each phase of the life cycle of the tick. Each assay comprised 10 treatment groups, consisting of one fungal isolate and one conidial concentration, and one control group containing only the vehicle of

  15. 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)

  16. Using in silico techniques: Isolation and characterization of an insect cuticle-degrading-protease gene from Beauveria bassiana.

    Science.gov (United States)

    Khan, Sehroon; Nadir, Sadia; Wang, Xuewen; Khan, Afsar; Xu, Jianchu; Li, Meng; Tao, Lihong; Khan, Siraj; Karunarathna, Samantha C

    2016-08-01

    Cuticle-degrading-proteases (CDPs) secreted by Beauveria spp. are pivotal biocontrol substances, possessing commercial potential for developing bio-pesticides. Therefore, a thoughtful and contemplative understanding and assessment of the structural and functional features of these proteases would markedly assist the development of biogenic pesticides. Computational molecular biology is a new facile alternative approach to the tedious experimental molecular biology; therefore, by using bioinformatics tools, we isolated and characterized an insect CDP gene from Beauveria bassiana 70 s.l. genomic DNA. The CDP gene (1240 bp with GeneBank accession no. KT804651.1) consisted of three introns and four CDS exons, and shared 74-100% sequence identity to the reference CDP genes. Its phylogenetic tree results showed a unique evolution pattern, and the predicted amino acid peptide (PAAP) consisted of 344 amino acid residues with pI, molecular weight, instability index, grand average hydropathicity value and aliphatic index of 7.2, 35.4 kDa, 24.45, -0.149, and 76.63, respectively. The gene possessed 74-89% amino acid sequence similarity to the 12 reference strains. Three motifs (Peptidase_S8 subtilase family) were detected in the PAAP, and the computed 3D structure possessed 79.09% structural identity to alkaline serine proteases. The PAAP had four (three serine proteases and one Pyridoxal-dependent decarboxylase) conserved domains, a disulfide bridge, two calcium binding sites, MY domain, and three predicted active sites in the serine family domains. These results will set the groundwork for further exploitation of proteases and understanding the mechanism of disease caused by cuticle-degrading-serine-proteases from entomopathogenic fungi.

  17. Antiproliferative and Apoptosis-Inducing Activities of 4-Isopropyl-2,6-bis(1-phenylethylphenol Isolated from Butanol Fraction of Cordyceps bassiana

    Directory of Open Access Journals (Sweden)

    Ji Hye Kim

    2015-01-01

    Full Text Available The Cordyceps species have been widely used for treating various cancer diseases. Although the Cordyceps species have been widely known as an alternative anticancer remedy, which compounds are responsible for their anticancer activity is not fully understood. In this study, therefore, we examined the anticancer activity of 5 isolated compounds derived from the butanol fraction (Cb-BF of Cordyceps bassiana. For this purpose, several cancer cell lines such as C6 glioma, MDA-MB-231, and A549 cells were employed and details of anticancer mechanism were further investigated. Of 5 compounds isolated by activity-guided fractionation from BF of Cb-EE, KTH-13, and 4-isopropyl-2,6-bis(1-phenylethylphenol, Cb-BF was found to be the most potent antiproliferative inhibitor of C6 glioma and MDA-MB-231 cell growth. KTH-13 treatment increased DNA laddering, upregulated the level of Annexin V positive cells, and altered morphological changes of C6 glioma and MDA-MB-231 cells. In addition, KTH-13 increased the levels of caspase 3, caspase 7, and caspase 9 cleaved forms as well as the protein level of Bax but not Bcl-2. It was also found that the phosphorylation of AKT and p85/PI3K was also clearly reduced by KTH-13 exposure. Therefore, our results suggest KTH-13 can act as a potent antiproliferative and apoptosis-inducing component from Cordyceps bassiana, contributing to the anticancer activity of this mushroom.

  18. Antiproliferative and Apoptosis-Inducing Activities of 4-Isopropyl-2,6-bis(1-phenylethyl)phenol Isolated from Butanol Fraction of Cordyceps bassiana

    Science.gov (United States)

    Kim, Ji Hye; Sung, Gi-Ho; Kim, Han Gyung; Park, Jae Gwang; Baek, Kwang-Soo; Yoon, Deok Hyo; Lee, Sang Yeol; Kang, Hyojeung; Song, Changsik; Cho, Jae Han; Lee, Kang-Hyo; Kim, Tae Woong

    2015-01-01

    The Cordyceps species have been widely used for treating various cancer diseases. Although the Cordyceps species have been widely known as an alternative anticancer remedy, which compounds are responsible for their anticancer activity is not fully understood. In this study, therefore, we examined the anticancer activity of 5 isolated compounds derived from the butanol fraction (Cb-BF) of Cordyceps bassiana. For this purpose, several cancer cell lines such as C6 glioma, MDA-MB-231, and A549 cells were employed and details of anticancer mechanism were further investigated. Of 5 compounds isolated by activity-guided fractionation from BF of Cb-EE, KTH-13, and 4-isopropyl-2,6-bis(1-phenylethyl)phenol, Cb-BF was found to be the most potent antiproliferative inhibitor of C6 glioma and MDA-MB-231 cell growth. KTH-13 treatment increased DNA laddering, upregulated the level of Annexin V positive cells, and altered morphological changes of C6 glioma and MDA-MB-231 cells. In addition, KTH-13 increased the levels of caspase 3, caspase 7, and caspase 9 cleaved forms as well as the protein level of Bax but not Bcl-2. It was also found that the phosphorylation of AKT and p85/PI3K was also clearly reduced by KTH-13 exposure. Therefore, our results suggest KTH-13 can act as a potent antiproliferative and apoptosis-inducing component from Cordyceps bassiana, contributing to the anticancer activity of this mushroom. PMID:25918546

  19. SELECTION OF BEAUVERIA BASSIANA (BALSAMO VUILLEMIN ISOLATES FOR MANAGEMENT OF MYZUS PERSICAE (SULTZAR (HOM.: APHIDAE BASED ON VIRULENCE AND GROWTH RELATED CHARACTERISTICS

    Directory of Open Access Journals (Sweden)

    Mohammad S. Al-alawi

    2014-01-01

    Full Text Available Isolates of the entomopathogenic fungus Beauveria bassiana originated from Jordan were evaluated for their efficacy against the green peach aphid, Myzus persicae under laboratory and greenhouse conditions. Efficacy evaluation involved testing all isolates at a concentration of 1×107 conidia/mL followed by concentration dependent and greenhouse bioassays for the top virulent isolates. Growth characteristics related to virulence were evaluated for high, intermediate and low virulent isolates. Results showed that three isolates namely: BAU004, BAU018 and BAU019 were highly virulent to the aphid in the laboratory causing more than 75% infection. In the greenhouse, the three isolates caused infection from 41.3 to 46.5%. For the growth characteristics, isolate BAU019 produced more spores than the other highly virulent ones including the commercial isolate GHA. Highly virulent isolates also showed faster hyphal growth than low virulent isolates. These findings indicate that isolates BAU004, Bau018 and BAU019 might be developed as commercial microbial insecticides for safe and effective control of green peach aphid.

  20. The pathogenicity of Beauveria bassiana: what happens after an endophytic phase in plants?

    Science.gov (United States)

    Akello, J; Dubois, T; Coyne, D; Kyamanywa, S

    2010-01-01

    The banana weevil Cosmopolites sordidus (Germar) (Coleoptera: Curculionidae) is a serious constraint to banana (Musa spp.) production throughout the world. The entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) offers a potential weevil management option, but conventional delivery mechanisms have limited its success. As an endophyte, however, B. bassiana can be efficiently delivered to banana planting materials for the potential management of C. sordidus. However, entomopathogens can change morphology and efficacy against their target host when successively sub-cultured on artificial media or when exposed to certain physical and chemical environmental conditions. Whether such changes occur in B. bassiana after an endophytic phase inside a banana plant remains unknown. The primary aim of our study was to evaluate the viability, growth, sporulation and pathogenicity of endophytic B. bassiana. To attain this, two sets of experiments, namely morphological characterization and larval bioassays, were conducted under laboratory conditions. In these experiments, growth and pathogenicity of the wild-type B. bassiana strain G41, obtained originally from banana farms, was compared with the endophytic B. bassiana strain G41, re-isolated from the rhizome of B. bassiana-inoculated banana plants at one month post-inoculation. Morphological characterization, conidial germination, colony growth and sporulation rate was assessed on SDAY media while pathogenicity was determined 15 days after immersing the larvae of C. sordidus in different conidial doses. No differences were observed in colony appearance and growth rate between the endophytic and wild-type strain. Percentage conidial germination for the endophytic strain (91.4-94.0%) was higher than for the wild-type (86.6-89.7%). LD50 equated 1.76 x 10(5) and 0.71 x 10(5) conidia/ml for the wild-type and endophytic B. bassiana strains, respectively, but did not differ between strains. Our study

  1. Molecular tracing of white muscardine in the silkworm, Bombyx mori (Linn.) II. Silkworm white muscardine is not caused by artificial release or natural epizootic of Beauveria bassiana in China.

    Science.gov (United States)

    Chen, Xue; Huang, Cui; He, Lingmin; Zhang, Shengli; Li, Zengzhi

    2015-02-01

    The fungal pathogen Beauveria bassiana causes serious economic losses in sericulture. Its origin is usually attributed to the release of B. bassiana insecticides against pine caterpillars (Dendrolimus punctuatus). In the present study, 488 B. bassiana isolates obtained from silkworm (Bombyx mori) collected from 13 Chinese provinces, and 327 B. bassiana isolates obtained from D. punctatus collected from 9 provinces, were analyzed for population genetic structure using the ISSR technique based on genetic distance. A UPGMA dendrogram clustered them into three independent clades: two B. mori clades and one D. punctatus clade. A 3-D principal component analysis further divided them into two completely independent host groups, revealing high host-specificity. This suggested that white muscardine occurring in B. mori populations throughout southern China was not caused by any B. bassiana strain either naturally prevailing in D. punctatus populations or by any strain artificially released as a fungal insecticide against D. punctatus. We further investigated the genetic differentiation coefficient Gst and gene flow between B. mori-pathogenic and D. punctatus-pathogenic B. bassiana isolates from across China and from five provinces inhabited by both B. mori and D. punctatus. The Gst value across China was computed as 0.410, while the values of the five provinces ranged from 0.508 to 0.689; all above 0.25, which is the threshold for significant genetic differentiation. This suggests that B. bassiana strains isolated from the two different hosts maintained their respective heredity without a convergent homogenization trend, and reduces the possibility that the host range of the caterpillar isolates could expand and enhance their virulence in B. mori. These findings indicate that the use of B. bassiana does not threaten the safety of sericulture.

  2. Caracterización molecular de 15 aislamientos de Beauveria bassiana asociados con Cosmopolites y Metamasius en plátano y banano en tres regiones de Colombia Molecular characterization of 15 isolations of Beauveria bassiana related to Cosmopolites and Metamasius in plantain and banana in three regions of Colombia

    Directory of Open Access Journals (Sweden)

    Diego Fernando Marmolejo

    2008-10-01

    Full Text Available Se colectaron picudos de Cosmopolites y Metamasius en municipios del Valle del Cauca, Caldas y Quindío. Se obtuvieron cultivos monospóricos con diluciones de 10-10 y 10-11. Los aislamientos fueron almacenados a -80°C con glicerol al 10% y el ADN a -20°C. Los marcadores moleculares RAM generaron 82 fragmentos de los cuales 67% fueron polimórficos con una heterocigocidad de 0.24, que indica diversidad media a alta. A un índice de similitud 0.84 se formaron 5 grupos: uno con 11 aislamientos y 4 con un solo aislamiento. En el gran grupo se detectó un duplicado y se encontró diversidad del hongo en los sitios muestreados. No se encontró relación entre aislamientos sobre Cosmopolites y Metamasius o zona geográfica en la formación de grupos genéticos.Weevils Cosmopolites and Metamasius in municipalities of the Valle del Cauca, Caldas and Quindío departaments of Colombia were collected. Monosporic cultures were obtained from10-10 and 10-11 dilutions of Beauveria bassiana. Isolates were kept at -80°C with 10% glycerol and DNA stored at -20°C. RAMs molecular markers generated a total of 82 fragments of which 67% were polymorphic. A heterozygosity value of 0.24 indicated a medium - high diversity. Five groups were formed which have a similarity value of 0.84 and one big group with 11 isolates and four groups with only one isolate. In the big group was detected a duplicate and fungi genetic diversity in the sampled places. Neither relationship among isolates of Cosmopolites and Metamasius nor geographical zone related to the formation of genetic groups.

  3. Molecular characterization of a novel amalgavirus from the entomopathogenic fungus Beauveria bassiana.

    Science.gov (United States)

    Koloniuk, Igor; Hrabáková, Lenka; Petrzik, Karel

    2015-06-01

    Beauveria bassiana is a ubiquitous entomopathogen infecting hundreds of insect species. We have determined the genomic organization and the complete nucleotide sequence of a novel virus isolated from the isolate A24 of B. bassiana. Phylogenetic analysis of the polymerase gene reveals that the virus, tentatively named Beauveria bassiana virus 1, belongs to the family Amalgaviridae and represents a distinct lineage of amalgaviruses infecting fungi.

  4. Infection of Helicoverpa armigera by endophytic Beauveria bassiana colonizing tomato plants

    Science.gov (United States)

    A novel endophytic strain of Beauveria bassiana was isolated from leaf tissue of a wild tomato plant. This strain and two B. bassiana strains previously isolated from soil were evaluated for their ability to endophytically colonize tomatoes and subsequent in planta efficacy against Helicoverpa armig...

  5. Bioconversion of 2‑Ethylpyridine by Beauveria bassiana

    OpenAIRE

    Parshikov, Igor A; Khasaeva, Fatima M

    2015-01-01

    Investigated the bioconversion of 2-ethylpyridine by the fungus Beauveria bassiana ATCC 7159. In the result of researches was obtained the hydroxylated derivative of the initial substrate. The yield of the product was observed as 60 %.

  6. 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.

  7. 球孢白僵菌九个菌株对赤拟谷盗成虫的毒效%Efficacy of nine isolates of Beauveria bassiana against adults of Tribolium castaneum (Coleoptera: Tenebrionidae)

    Institute of Scientific and Technical Information of China (English)

    Hediyeh GOLSHAN; Moosa SABER; Farzad MAJIDI-SHILSAR; Farrokh KARIMI

    2013-01-01

    Beauveria bassiana (Balsamo) Vuillemin is one of the most important entomopathogenic fungi and has been widely used for many insect pests all over the world.In this study,the pathogenicity of nine isolates of B.bassiana was evaluated against adults of Tribolium castaneum (Herbst).Fifteen adults of the pest were submerged into the four suspensions (1 × 106,1 × 107,1 × 10s and 1 × l09 conidia/mL) of different isolates of B.bassiana for 20 s,and the mortality was recorded daily for 14 d.Results showed that IRAN 440C and IRAN 187C isolates had the lowest (5.04 × 107 conidia/mL) and highest (5.05 ×108 conidia/mL) LCs0,and DEBI 005 and DEBI 014 isolates had the shortest (2.88 d) and longest (4.96 d) LT50,respectively.According to the LC5o and LT50 values and the mortality,IRAN 440C is a perfect isolate for using in controlling the pest.%球孢白僵菌Beauveria bassiana (Balsamo) Vuillemin是最重要的昆虫病原真菌,广泛用于防治世界各地的多种害虫.本研究评价了球孢白僵菌9个菌株对赤拟谷盗Tribolium castaneum (Herbst)成虫的致病性.将15头赤拟谷盗成虫浸入到4个浓度(1×106,l×107,l×lo8和l×109个分生孢子/mL)的白僵菌菌株中20 s,14 d内每日记录成虫的死亡率.结果表明:IRAN 440C菌株对赤拟谷盗成虫的LC50最低(5.04 ×l07个分生孢子/mL),IRAN187C菌株的最高(5.05 ×lo8个分生孢子/mL);DEBI 005菌株对赤拟谷盗成虫的LT5o最短(2.88 d),DEBI 014菌株的最长(4.96 d).根据LC50,LT50和死亡率结果得出IRAN 440C是防治这一害虫的理想菌株.

  8. 4-Isopropyl-2,6-bis(1-phenylethylaniline 1, an Analogue of KTH-13 Isolated from Cordyceps bassiana, Inhibits the NF-κB-Mediated Inflammatory Response

    Directory of Open Access Journals (Sweden)

    Woo Seok Yang

    2015-01-01

    Full Text Available The Cordyceps species has been a good source of compounds with anticancer and anti-inflammatory activities. Recently, we reported a novel compound (4-isopropyl-2,6-bis(1-phenylethylphenol, KTH-13 with anticancer activity isolated from Cordyceps bassiana and created several derivatives to increase its pharmacological activity. In this study, we tested one of the KTH-013 derivatives, 4-isopropyl-2,6-bis(1-phenylethylaniline 1 (KTH-13-AD1, with regard to anti-inflammatory activity under macrophage-mediated inflammatory conditions. KTH-13-AD1 clearly suppressed the production of nitric oxide (NO and reactive oxygen species (ROS in lipopolysaccharide (LPS and sodium nitroprusside- (SNP- treated macrophage-like cells (RAW264.7 cells. Similarly, this compound also reduced mRNA expression of inducible NO synthase (iNOS and tumor necrosis factor-α (TNF-α, as analyzed by RT-PCR and real-time PCR. Interestingly, KTH-13-AD1 strongly diminished NF-κB-mediated luciferase activities and nuclear translocation of NF-κB family proteins. In accordance, KTH-13-AD1 suppressed the upstream signaling pathway of NF-κB activation, including IκBα, IKKα/β, AKT, p85/PI3K, and Src in a time- and dose-dependent manner. The autophosphorylation of Src and NF-κB observed during the overexpression of Src was also suppressed by KTH-13-AD1. These results strongly suggest that KTH-13-AD1 has strong anti-inflammatory features mediated by suppression of the Src/NF-κB regulatory loop.

  9. Determinação da CL90 e TL90 do isolado IBCB66 de Beauveria bassiana (Ascomycetes: Clavicipitaceae para o controle de Rhipicephalus (Boophilus microplus (Acari: Ixodidae Determination of LC 90 and LT 90 of IBCB66 Beauveria bassiana (Ascomycetes: Clavicipitaceae isolate for Rhipicephalus (Boophilus microplus (Acari: Ixodidae control

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    Barci Leila A. G.

    2009-12-01

    Full Text Available O presente trabalho teve por objetivo avaliar a patogenicidade e a virulência do isolado IBCB66 de Beauveria bassiana para larvas de Rhipicephalus (Boophilus microplus. O isolado IBCB66 foi utilizado como padrão, com a finalidade de determinar a CL50 (Concentração Letal, CL90, TL50 (Tempo Letal e TL90. O isolado IBCB66 foi testado em seis concentrações diferentes (5 × 10(6, 10(7, 5 × 10(7, 10(8, 5 × 10(8 e 10(9 para determinar a porcentagem de mortalidade. A mortalidade total de larvas foi observada 18 dias após o início do teste no grupo tratado com 5 × 10(9 conídios.mL-1. A análise de Probit dos dados consignados apontou a CL50 e CL90 concentrações de 3 × 10(7 e 5 × 10(8 conídios.mL-1 e para TL50 e TL90, foram 10 e 16 dias, respectivamente.The objective of this research was to evaluate the pathogenicity and the virulence of the IBCB66 isolate of Beauveria bassiana on infected larvae of Rhipicephalus (Boophilus microplus. The IBCB66 fungus strain was used as standard isolates of B. bassiana against R. (B. microplus larvae. The larval bioassay tests using the IBCB66 isolate were carried out to determine the (Lethal Concentration LC50, LC90, (Lethal Time LT50 and LT90. The IBCB66 fungus strain was tested at six different concentrations (5 × 10(6, 10(7, 5 × 10(7, 10(8, 5 × 10(8 and 10(9 conidia.mL-1 to determine the percentage of larval mortality. In addition, a Probit analysis was also performed. Total larval mortality was observed eighteen days after the beginning of the test in the group treated with 5 × 10(9 conidia.mL-1. The LC50 and LC90 were 3 × 10(7 and 5 × 10(8 respectively and the LT50 and LT90 were 10 and 16 days.

  10. Lethal effects of a Mexican Beauveria bassiana (Balsamo strain against Meccus pallidipennis (Stal

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    Zumaquero Rios José Lino

    2014-06-01

    Full Text Available The entomopathogenic fungus Beauveria bassiana (Balsamo 1835 Vuillemin is an effective alternative control agent against some agricultural pests and biological vectors of important diseases such as Chagas disease. In this work we studied an isolate of Beauveria bassiana from of the town of San Antonio Rayón, Puebla, Mexico and its entomopathogenic effects on Meccus pallidipennis (Stal 1872. Phylogenetic analysis using molecular comparison of the ITS and EF1α genes, showed that the resulting cladogram places the BUAP 04 strain with a relationship closer to the AFAO 9-6 strain, within the diversity of the B. bassiana sensu lato group. Although there was the possibility that BUAP 04 strain was a direct descendant of strains used in campaigns of biologic control, molecular study allowed us to recognize that it was a different fungus due to numerous inserts. A strain isolated from a T. dimiata was evaluated for pathogenicity against another triatoma (Meccus pallidipennis species obtaining an LC50 of 4.16 x 10(6 spores/mL, confirming that the BUAP 04 strain is virulent for M. pallidipennis and could be a good prospect for formulations to control M. pallidipennis.

  11. Lethal effects of a Mexican Beauveria bassiana (Balsamo) strain against Meccus pallidipennis (Stal)

    Science.gov (United States)

    Lino, Zumaquero Rios José; Juventino, López-Tlacomulco José; Raúl, Rojas García; Estibaliz, Sansinenea

    2014-01-01

    The entomopathogenic fungus Beauveria bassiana (Balsamo 1835) Vuillemin is an effective alternative control agent against some agricultural pests and biological vectors of important diseases such as Chagas disease. In this work we studied an isolate of Beauveria bassiana from of the town of San Antonio Rayón, Puebla, Mexico and its entomopathogenic effects on Meccus pallidipennis (Stal 1872). Phylogenetic analysis using molecular comparison of the ITS and EF1α genes, showed that the resulting cladogram places the BUAP 04 strain with a relationship closer to the AFAO 9-6 strain, within the diversity of the B. bassiana sensu lato group. Although there was the possibility that BUAP 04 strain was a direct descendant of strains used in campaigns of biologic control, molecular study allowed us to recognize that it was a different fungus due to numerous inserts. A strain isolated from a T. dimiata was evaluated for pathogenicity against another triatoma (Meccus pallidipennis) species obtaining an LC50 of 4.16 × 106 spores/mL, confirming that the BUAP 04 strain is virulent for M. pallidipennis and could be a good prospect for formulations to control M. pallidipennis. PMID:25242941

  12. Lethal effects of a Mexican Beauveria bassiana (Balsamo) strain against Meccus pallidipennis (Stal).

    Science.gov (United States)

    Lino, Zumaquero Rios José; Juventino, López-Tlacomulco José; Raúl, Rojas García; Estibaliz, Sansinenea

    2014-01-01

    The entomopathogenic fungus Beauveria bassiana (Balsamo 1835) Vuillemin is an effective alternative control agent against some agricultural pests and biological vectors of important diseases such as Chagas disease. In this work we studied an isolate of Beauveria bassiana from of the town of San Antonio Rayón, Puebla, Mexico and its entomopathogenic effects on Meccus pallidipennis (Stal 1872). Phylogenetic analysis using molecular comparison of the ITS and EF1α genes, showed that the resulting cladogram places the BUAP 04 strain with a relationship closer to the AFAO 9-6 strain, within the diversity of the B. bassiana sensu lato group. Although there was the possibility that BUAP 04 strain was a direct descendant of strains used in campaigns of biologic control, molecular study allowed us to recognize that it was a different fungus due to numerous inserts. A strain isolated from a T. dimiata was evaluated for pathogenicity against another triatoma (Meccus pallidipennis) species obtaining an LC50 of 4.16 × 10(6) spores/mL, confirming that the BUAP 04 strain is virulent for M. pallidipennis and could be a good prospect for formulations to control M. pallidipennis.

  13. Studies on the Cloning and Expression of Bbchit1 Gene of Beauveria bassiana NCIM 1216.

    Science.gov (United States)

    Pinnamaneni, Rajasekhar; Kalidas, P; Sambasiva Rao, K R S

    2011-07-01

    Beauveria bassiana is a biocontrol agent which shows entomopathogenecity on insect pests especially the Lepidopterons invading the agriculturally important crops. The mode of infection is through the cuticle by degrading the chitin present on it which is enabled by the exochitinase enzyme of Bbchit1 gene. A good quality genomic DNA was isolated from Beauveria bassiana NCIM 1216 and amplified with specific primers to isolate the gene corresponding to Bbchit1 which codes for the exochitinase enzyme that is responsible for pathogenesis. The Bbchit1 gene of B. bassiana was transformed with the binary plasmid pBANF-bar-pAN-Bbchit1, in which the Bbchit1 gene was placed downstream of the constitutive gpd promoter, which was mediated by A. tumefaciens, and transformants were selected on the basis of herbicide resistance. Fifty herbicide resistant colonies were obtained and analyzed. The exochitinase produced by these transformants was observed maximum on the 7th day of inoculation in both which was 0.09 μmol/ml/min for the purified fraction and 0.06 μmol/ml/min for the crude extract. The chitinolytic activity was observed maximum at pH 5 and at temperature of 40°C. The genetically modified pure form can be used in the production of transgenic plants and in bringing out commercial formulation for the control of Lepidopteran pests.

  14. Isolation and Identification of highly pathogenic Beauveria bassiana against Thrips%对蓟马类害虫高致病性球孢白僵菌的分离、鉴定

    Institute of Scientific and Technical Information of China (English)

    王俊平; 郑长英

    2011-01-01

    12 different entomopathogenic fungi isolated from diseased Drosicha corpulenta Kuwana showed high virulence with 70%~90% mortality. CYT10 strain was identified based on the morphological and molecular characteristics. Conidiogenous cells were short-globose to flask-shaped and the conidiophore elongated in a long zig-zag extension after each conidium is produced. The result of Blast search indicated that the ITS and EF1-α sequence of CYT10 showed a high similarity with those of other strain of Beauveria bassiana (>95%). Phylogenetic analysis showed that it was closely related to ARSEF-1185 strain, which was belonging to B. Bassiana. CYT10 strain was classified as a member of the species B. Bassiana based on the morphological characteristics and analysis of DNA sequences.%从草履蚧僵虫体内分离出12株病原真菌,对蓟马类昆虫具有70%~90%的致死率.现选出一株病原真菌CYT10利用形态和分子的特点进行鉴定.扫描电镜下观察,该菌产孢细胞簇生瓶状,“之”型产孢轴.经PCR获得rDNA-ITS和EF1 -a核苷酸序列,与其他球孢白僵菌的相应序列一致性达95%以上,进化分析CYT10菌株与球孢白僵菌Beauveria bassiana聚为一类.根据菌株的形态观察和DNA序列分析确定该菌为球孢白僵菌.

  15. 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.

  16. 玉米根际土球孢白僵菌(Beauveria bassiana)的分离与鉴定%Isolation and Identification of Beauveria bassiana From Maize Rhizosphere Soil

    Institute of Scientific and Technical Information of China (English)

    陈方新; 梅玉云; 张强; 齐永霞; 丁婷

    2016-01-01

    为明确虫生真菌在玉米根际的分布情况,通过选择性培养基对采自安徽省涡阳、萧县、蒙城3地的玉米根际土壤中的虫生真菌进行了分离,并结合形态特征及rDNA ITS序列分析对分离的菌株进行了鉴定.结果表明,从玉米根际土壤中共分离得到39个白僵菌菌株,经鉴定均为球孢白僵菌(Beauveria bassiana),且不同菌株间的菌落生长速度和分生孢子产生量存在显著差异.本研究对丰富玉米地下害虫及土传病害的生物防治资源具有重要的意义.

  17. 银杏大蚕蛾白僵菌的分离、培养及应用试验%Isolation,culture and application of Beauveria bassiana for the control of Dictyoploca japonica

    Institute of Scientific and Technical Information of China (English)

    杨世璋; 吴猛耐; 陈杰; 林琳; 付成松; 杜友兵; 蔡泽荣

    2001-01-01

    从银杏大蚕蛾僵死虫(蛹)尸上分离的白僵菌B21.1,经过室内扩大培养,生产出含孢量185亿孢子/g菌剂,以其2亿孢子/ml菌液对银杏大蚕蛾幼虫室内毒力测定,表现出很高的致病能力:4天死亡率近半,10天校正死亡率达到98.51%;林间喷雾防治,10天左右致病死亡近50%,21天校正死亡率为84.36%;与其它菌株(剂)试验比较,其致病死亡率和杀虫效果均明显高于其他各供试菌株,表现了较强的专化性。%Five isolates of Beauveria bassiana from different sources were examined for pathogenicity to Dictyoploca japonica. Compared with other four isolates,the isolates B21.1 derived from dead body of Dictyoploca japonica showed the best effectiveness.Two inoculation experiments,one in lab and the other in the field,were carried out.The mortality of pests caused by isolates B21.1 come to 98.51% in lab 10 days after inoculation and 84.36% in the field 21 days after inoculation.The results suggest that isolate of Beauveria bassiana from the host itself shows good prospect for the control of Dictyoploca japonica.

  18. Variation in virulence of Beauveria bassiana and B. pseudobassiana to the pine weevil Pissodes nemorensis in relation to mycelium characteristics and virulence genes.

    Science.gov (United States)

    Romón, Pedro; Hatting, Hardus; Goldarazena, Arturo; Iturrondobeitia, Juan Carlos

    2017-02-01

    Entomopathogenic fungi such as Beauveria spp. have potential applications in the biocontrol of insect pests but little is known regarding their infectivity to the pine weevil Pissodes nemorensis. In this study, five isolates of Beauveria pseudobassiana and five isolates of Beauveria bassiana were tested for characteristics correlating with virulence on P. nemorensis. Isolate UAMH301 had the lowest mean lethal concentration value whereas the highest value was obtained with isolate LRC137. Growth rate was negatively correlated with virulence in B. bassiana, because isolate LRC137, the least virulent isolate, grew much more rapidly than the other B. bassiana isolates on SDYA. In contrast, its growth on a hyperosmotic medium was the slowest. Sporulation rate and conidial area were not correlated with virulence. Mycelial cell density was positively correlated with virulence in both species, and the four tested genes appear to be one-copy genes. Bbchit1 and Bbhog1, genes respectively encoding a chitinase and a protein kinase, induced relative expression levels were positively correlated with virulence in B. pseudobassiana. We discuss in terms of previous morphological, physiological and genetic parameters related to virulence in Beauveria and the importance of testing the expression of putative virulence genes in comparison with their basal transcript levels.

  19. Discovery and demonstration of the teleomorph of Beauve-ria bassiana (Bals.) Vuill. , an important entomogenous fun-gus

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A Cordyceps specimen was collected in Anhui, China, a strain of Beauveria bassiana, an important entomopathogenic fungus for biological pest control, was isolated and their relationship was demonstrated by microcycle conidiation. The teleomorph is an undescribed species and is named Cordyceps bassiana.

  20. Tyrosinase Inhibition Type of Isolated Compounds Obtained from Pachyrhizus erosus

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    Endang Lukitaningsih

    2013-12-01

    Full Text Available In Indonesia, Bengkoang (Phacyrhizus erosus have been used as one of cosmetics especially as sun screening and skin whitening materials. Six active compounds in Bengkoang with antioxidant and skin whitening activities have been isolated, namely daidzein, daidzin, genistin, (8,9-furanyl-pterocarpan-3-ol, 4-(2-(furane-2-ylethyl-2-methyl-2,5-dihydro-furane-3-carbaldehyde and 2-butoxy-2,5-bis(hydroxymethyl-tetrahydrofurane-3,4-diol. According to literatures, the type of their tyrosinase inhibitory activity has not yet reported. The determination of whitening activity of each compound was evaluated by the evaluation of Lineweaver-Burk plot. The result showed that five compounds had competitive inhibitory activity and 8,9-furanyl-pterocarpan-3-ol showed a non-competitive inhibition.

  1. 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

    Directory of Open Access Journals (Sweden)

    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.

  2. Screening of liquid media and fermentation of an endophytic Beauveria bassiana strain in a bioreactor.

    Science.gov (United States)

    Lohse, Rieke; Jakobs-Schönwandt, Desiree; Patel, Anant V

    2014-01-01

    A novel approach for biological control of insect pests could be the use of the endophytic entomopathogenic Beauveria bassiana isolate ATP-02. For the utilization of the endophyte as a commercial biocontrol agent, the fungus has to be mass-produced. B. bassiana was raised in shake flask cultures to produce high concentrations of total spores (TS), which include blastospores (BS) and submerged conidiospores (SCS). The highest concentration of 1.33×10(9) TS/mL and the highest yield of 5.32×10(10) TS/g sucrose was obtained in the TKI broth with 5% sugar beet molasses which consists of 50% sucrose as a carbon source. In spite of the lower sugar concentration (2.5%) the amount of TS could be increased up to 11-times in contrast to the cultivation with 5% sucrose. The scale-up to a 2 L stirred tank reactor was carried out at 25°C, 200-600 rpm and 1 vvm at pH 5.5. A TS yield of 5.2×10(10) TS/g sucrose corresponding to a SCS yield of 0.2×10(10) SCS/g sucrose was obtained after 216 h. With regards to the culture medium the cost of 10(12) TS amounts to 0.24 €. Plutella xylostella larvae, which were fed with oilseed rape leaves treated with spores from fermentation resulted in 77 ± 5% mortality. Moreover, spores from submerged cultivation were able to colonize oilseed rape leaves via leaf application. This is the first report of fermentation of an endophytic B. bassiana strain in a low-cost culture medium to very high yields of TS.

  3. THE INFLUENCE OF ULTRAVIOLET LIGHT ON PATHOGENICITY OF ENTOMOPATHOGENIC FUNGUS BEAUVERIA BASSIANA (BALSAMO VUILLEMIN TO THE EUROPEAN CORN BORER, OSTRINIA NUBILALIS HBN. (LEPIDOPTERA: CRAMBIDAE

    Directory of Open Access Journals (Sweden)

    L Cagán

    2002-05-01

    Full Text Available The influence of different doses of ultraviolet (UV light on the pathogenicity of entomopathogenic fungus Beauveria bassiana (Balsamo Vuillemin to the European corn borer, Ostrinia nubilalis Hbn., and radial growth of fungus was studied in laboratory conditions. The suspensions of B. bassiana isolate SK99 were exposed to UV light. Four different doses of UV light were used in the experiment. The distance between exposed suspensions and UV light source was 0.3 m. Exposure duration was 15, 30, 45 and 60 minutes (as A, B, C and D variants. Control variant SK99 and obtained variants SK99A, SK99B, SK99C and SK99D were cultivated 21 days on Sabourard-dextrose agar. The larvae of O. nubilalis were infected with dry powder consisted of mycelia and spores from fungus cultures. During 10 days, the mortality of infected larvae was evaluated. It was ascertained that UV light exposition significantly influenced the mortality effect of B. bassiana isolates to O. nubilalis larvae. Variant SK99C showed the highest level of infectivity. Radial growth of UV variants was slower with rising time of exposure. The best ability to grow possessed non-irradiated isolate SK99 and the worse variant SK99D. The difference between these two variants was significant.

  4. BIOLOGICAL CHARACTERISTICS OF THREE ISOLATES OF BEAUVERIA BASSIANA AND THEIR VIRULANCE AGAINST WHEAT APHIDS%球孢白僵菌的生物学特性及对小麦蚜虫的毒力

    Institute of Scientific and Technical Information of China (English)

    张永军; 王中康; 殷幼平; 裴炎

    2001-01-01

    Three isolates of Beauveria bassiana, Bb 02, Bb 03 and Bb 07, were cultured at different temperature and pH and on different media. The optimum temperature for their growth ranged from 25℃ to 30℃ and Bb 02 and Bb 07 grew well at pH 4.0 ~ 6.0. The addition of peptone and yeast extract stimulated their growth,while glucose and maltose had little promoting effect. All the three isolmes grew well on the media PPDA,SDAY and SMAY. Bioassay showed that they were strongly virulent against whest aphids,though their optimum dosage varied a little. The opimum concentration was 106/mL for Bb 02,107/mL for Bb 03 and 107 ~ 108/mL for Bb 07.%3株球孢白僵茵(Beauveria bassiana)Bb 02,Bb 03和Bb 07在不同温度,pH值和培养基上培养,结果表明,三者的最适生长温度在25℃~30℃间;Bb 02和Bb 07在pH4.0~6.0生长良好;适量的蛋白胨或酵母浸膏可促进菌株的生长,但对葡萄糖和麦芽糖的利用能力没有明显差异,三者在PPDA,SDAY和SMAY上生长良好。生物测定表明,3茵株对小麦蚜虫均具有较强的毒力,但各菌株的最适剂量略有差异,其中Bb 02以106mL-1较佳,Bb 03以107mL-1最好,Bb07则介于107mL-1~108mL-1之间。

  5. Field evaluation of the synergistic effects of neem oil with Beauveria bassiana (Hypocreales: Clavicipitaceae) and Bacillus thuringiensis var. kurstaki (Bacillales: Bacillaceae)

    NARCIS (Netherlands)

    Togbe, C.E.; Zannou, E.; Gbehounou, G.; Kossou, D.; Huis, van A.

    2014-01-01

    In the present study, the synergistic effects of Beauveria bassiana (Bals.-Criv. Vuill.) (isolate Bb11) and Bacillus thuringiensis var. kurstaki (Berliner) with neem oil were evaluated in three agroecological zones in Be´nin. Four bioinsecticide treatments (neem oil, neem oil and B. bassiana used

  6. Field evaluation of the synergistic effects of neem oil with Beauveria bassiana (Hypocreales: Clavicipitaceae) and Bacillus thuringiensis var. kurstaki (Bacillales: Bacillaceae)

    NARCIS (Netherlands)

    Togbe, C.E.; Zannou, E.; Gbehounou, G.; Kossou, D.; Huis, van A.

    2014-01-01

    In the present study, the synergistic effects of Beauveria bassiana (Bals.-Criv. Vuill.) (isolate Bb11) and Bacillus thuringiensis var. kurstaki (Berliner) with neem oil were evaluated in three agroecological zones in Be´nin. Four bioinsecticide treatments (neem oil, neem oil and B. bassiana used se

  7. Cultural Characteristics and Fruiting Body Production in Cordyceps bassiana

    Science.gov (United States)

    Lee, Je-O; Shrestha, Bhushan; Sung, Gi-Ho; Han, Sang-Kuk; Kim, Tae-Wong

    2010-01-01

    Single ascospore isolates of Cordyceps bassiana were observed for their colony pigmentation on Sabouraud Dextrose agar plus Yeast Extract (SDAY) plates and were inoculated in a brown rice medium for production of fruiting bodies. Colony pigmentation did not show any relationship with perithecial stromata formation. The isolates were also grown on opposite sides of SDAY agar plates and were observed for vegetative compatibility. Neither vegetative compatibility nor perithecial stromata could be found to be related to each other. It was concluded that fertile fruiting body production was independent of colony pigmentation and vegetative compatibility. Synnemata formation was found to be more common than perithecial stromata formation. This might be due to its highly conidiogenous anamorphic stage, i.e., Beauveria bassiana. PMID:23956638

  8. Beauveria bassiana as an endophyte

    DEFF Research Database (Denmark)

    McKinnon, Aimee C.; Saari, Susanna; Moran-Diez, Maria E.;

    2017-01-01

    In the last decade there has been increased focus on the potential of endophytic Beauveria bassiana for the biocontrol of insect herbivores. Generally, detection of endophytes is acknowledged to be problematic and recovery method-dependent. Herein, we critically analyse the methodology reported...

  9. Mississippi Delta native strain of Beauveria bassiana for control of TPB

    Science.gov (United States)

    Previous works reported that Beauveria bassiana (Balsamo) isolates from tarnished plant bug, TPB, Lygus lineolaris Palisot de Beauvois, (Hemiptera: Miridae) have shown a great potential to be used as a microbial control for TPB, particularly the NI8 strain, isolated from the TPB from Mississippi De...

  10. 4-Isopropyl-2,6-bis(1-phenylethyl)aniline 1, an Analogue of KTH-13 Isolated from Cordyceps bassiana, Inhibits the NF-κB-Mediated Inflammatory Response

    Science.gov (United States)

    Yang, Woo Seok; Ratan, Zubair Ahmed; Kim, Gihyeon; Lee, Yunmi; Kim, Mi-Yeon; Kim, Jong-Hoon; Cho, Jae Youl

    2015-01-01

    The Cordyceps species has been a good source of compounds with anticancer and anti-inflammatory activities. Recently, we reported a novel compound (4-isopropyl-2,6-bis(1-phenylethyl)phenol, KTH-13) with anticancer activity isolated from Cordyceps bassiana and created several derivatives to increase its pharmacological activity. In this study, we tested one of the KTH-013 derivatives, 4-isopropyl-2,6-bis(1-phenylethyl)aniline 1 (KTH-13-AD1), with regard to anti-inflammatory activity under macrophage-mediated inflammatory conditions. KTH-13-AD1 clearly suppressed the production of nitric oxide (NO) and reactive oxygen species (ROS) in lipopolysaccharide (LPS) and sodium nitroprusside- (SNP-) treated macrophage-like cells (RAW264.7 cells). Similarly, this compound also reduced mRNA expression of inducible NO synthase (iNOS) and tumor necrosis factor-α (TNF-α), as analyzed by RT-PCR and real-time PCR. Interestingly, KTH-13-AD1 strongly diminished NF-κB-mediated luciferase activities and nuclear translocation of NF-κB family proteins. In accordance, KTH-13-AD1 suppressed the upstream signaling pathway of NF-κB activation, including IκBα, IKKα/β, AKT, p85/PI3K, and Src in a time- and dose-dependent manner. The autophosphorylation of Src and NF-κB observed during the overexpression of Src was also suppressed by KTH-13-AD1. These results strongly suggest that KTH-13-AD1 has strong anti-inflammatory features mediated by suppression of the Src/NF-κB regulatory loop. PMID:26819495

  11. Effect of surfactants and temperature on germination and vegetative growth of Beauveria bassiana.

    Science.gov (United States)

    Mwamburi, Lizzy A; Laing, Mark D; Miller, Ray M

    2015-03-01

    Three non-ionic surfactants: Tween20, Tween80 and Breakthru (®) were screened for their effects on spore germination and mycelial growth rates and for their influence on three isolates of Beauveria bassiana spore germination at various temperatures. Tween20 and Tween80 were compatible with all the B. bassiana isolates in the germination studies, but inhibited germination at higher surfactant concentrations, irrespective of the conidial concentrations . Breakthru (®) had an inhibitory effect on germination even at the lowest concentration of 0.1% on all the B. bassiana isolates. The effects of the surfactants on spore germination did not correspond with their effects on colony growth. Conidial viability within the same formulation declined significantly with increases in temperature, irrespective of the surfactant. The optimal temperature for conidial germination of B. bassiana isolates was approximately 25 °C with an upper limit at 30 °C. Isolate 7320 was identified as the least affected by the different surfactants. This isolate was able to germinate rapidly in a broad temperature range of 25-30 °C after 24 h, this characteristic being an essential factor in controlling house fly populations in poultry houses.

  12. Adhesion of the Entomopathogenic Fungus Beauveria (Cordyceps) bassiana to Substrata†

    Science.gov (United States)

    Holder, Diane J.; Keyhani, Nemat O.

    2005-01-01

    The entomopathogenic fungus Beauveria bassiana produces at least three distinct single-cell propagules, aerial conidia, vegetative cells termed blastospores, and submerged conidia, which can be isolated from agar plates, from rich broth liquid cultures, and under nutrient limitation conditions in submerged cultures, respectively. Fluorescently labeled fungal cells were used to quantify the kinetics of adhesion of these cell types to surfaces having various hydrophobic or hydrophilic properties. Aerial conidia adhered poorly to weakly polar surfaces and rapidly to both hydrophobic and hydrophilic surfaces but could be readily washed off the latter surfaces. In contrast, blastospores bound poorly to hydrophobic surfaces, forming small aggregates, bound rapidly to hydrophilic surfaces, and required a longer incubation time to bind to weakly polar surfaces than to hydrophilic surfaces. Submerged conidia displayed the broadest binding specificity, adhering to hydrophobic, weakly polar, and hydrophilic surfaces. The adhesion of the B. bassiana cell types also differed in sensitivity to glycosidase and protease treatments, pH, and addition of various carbohydrate competitors and detergents. The outer cell wall layer of aerial conidia contained sodium dodecyl sulfate-insoluble, trifluoroacetic acid-soluble proteins (presumably hydrophobins) that were not present on either blastospores or submerged conidia. The variations in the cell surface properties leading to the different adhesion qualities of B. bassiana aerial conidia, blastospores, and submerged conidia could lead to rational design decisions for improving the efficacy and possibly the specificity of entomopathogenic fungi for host targets. PMID:16151112

  13. Antimicrobial Resistance among Campylobacter Isolates Obtained from Retail Chicken Meat and Offal Products in Japan

    National Research Council Canada - National Science Library

    Hidano, Arata; Yamamoto, Takehisa; Hayama, Yoko; Muroga, Norihiko; Kobayashi, Sota; Nishida, Takeshi; Tsutsui, Toshiyuki

    2014-01-01

    .... In this study, we aimed to demonstrate the overall trend in antimicrobial resistance among Campylobacter isolates obtained from chicken meat and offal products collected from a wide geographic area throughout Japan...

  14. Comparative genomics of Beauveria bassiana: uncovering signatures of virulence against mosquitoes.

    Science.gov (United States)

    Valero-Jiménez, Claudio A; Faino, Luigi; Spring In't Veld, Daphne; Smit, Sandra; Zwaan, Bas J; van Kan, Jan A L

    2016-12-01

    Entomopathogenic fungi such as Beauveria bassiana are promising biological agents for control of malaria mosquitoes. Indeed, infection with B. bassiana reduces the lifespan of mosquitoes in the laboratory and in the field. Natural isolates of B. bassiana show up to 10-fold differences in virulence between the most and the least virulent isolate. In this study, we sequenced the genomes of five isolates representing the extremes of low/high virulence and three RNA libraries, and applied a genome comparison approach to uncover genetic mechanisms underpinning virulence. A high-quality, near-complete genome assembly was achieved for the highly virulent isolate Bb8028, which was compared to the assemblies of the four other isolates. Whole genome analysis showed a high level of genetic diversity between the five isolates (2.85-16.8 SNPs/kb), which grouped into two distinct phylogenetic clusters. Mating type gene analysis revealed the presence of either the MAT1-1-1 or the MAT1-2-1 gene. Moreover, a putative new MAT gene (MAT1-2-8) was detected in the MAT1-2 locus. Comparative genome analysis revealed that Bb8028 contains 163 genes exclusive for this isolate. These unique genes have a tendency to cluster in the genome and to be often located near the telomeres. Among the genes unique to Bb8028 are a Non-Ribosomal Peptide Synthetase (NRPS) secondary metabolite gene cluster, a polyketide synthase (PKS) gene, and five genes with homology to bacterial toxins. A survey of candidate virulence genes for B. bassiana is presented. Our results indicate several genes and molecular processes that may underpin virulence towards mosquitoes. Thus, the genome sequences of five isolates of B. bassiana provide a better understanding of the natural variation in virulence and will offer a major resource for future research on this important biological control agent.

  15. Laboratory invesitgations of variability of Ascochyta fabae Speg. isolates obtained from horse bean

    Directory of Open Access Journals (Sweden)

    Antoni Józef Filipowicz

    2014-08-01

    Full Text Available Fifty five isolates of Ascochyta fabae Speg. were investigated. They were selected from 1650 isolates of this fungus obtained from horse bean seeds in 1974-1976. All the isolates grew and sporulated on Potato Dextrose Agar, Malt Agar and Horse Bean Agar. The rate of their growth amounted to 1-4 mm per 24 hours. The variability of isolates in size of pycnidia and conidia and number of sepia was noticed. A few spores with untypical shapes were observed as well.

  16. Soil application of Beauveria bassiana GHA against apple sawfly, Hoplocampa testudinea (Hymenoptera: Tenthredinidae)

    DEFF Research Database (Denmark)

    Świergiel, Weronika; Meyling, Nicolai Vitt; Porcel, Mario;

    2016-01-01

    Low impact alternatives to synthetic insecticides for the control of apple sawfly (Hoplocampa testudinea Klug) are scarce encumbering pest management in organic apple orchards. We investigated the soil persistence and field efficacy of the entomopathogenic fungus Beauveria bassiana (Balsamo......) Vuillemin (BotaniGard) against apple sawfly under common organic orchard practices. We also assessed the efficacy of B. bassiana GHA and Metarhizium brunneum Petch (indigenous strain) against sawfly in the laboratory. Larvae treated with either fungus in the laboratory died faster than control larvae...... and displayed 49.4%-68.4% mycosis. In the field, B. bassiana density remained high in the week after application, during larval descent to the soil. Fungal density decreased to 25% at 49 d after application and to 0.4% after 55 weeks. Molecular markers revealed that the majority of fungal isolates recovered...

  17. Assessments of the microbial biocontrol potential of novel strains of Beauveria bassiana s.l. against greenhouse shore fly, Scatella tenuicosta: virulence, mass production capacity, and effects on shore fly reproduction

    Science.gov (United States)

    The biological control potential of three strains of Beauveria bassiana s.l. originally isolated from the shore fly Scatella ternuicosta was assessed in a series of laboratory bioassays. Comparisons were made to two commercially available strains of B. bassiana. Two of the new strains proved 27–67 t...

  18. Effects of indole-3-acetic acid on Botrytis cinerea isolates obtained from potted plants.

    Science.gov (United States)

    Martínez, J A; Valdés, R; Gómez-Bellot, M J; Bañón, S

    2011-01-01

    We study the growth of different isolates of Botrytis cinerea collected from potted plants which were affected by Botrytis blight in southern Spain during recent years. These isolates, which show widely phenotypic differences when grown in vitro, are differentially affected by growth temperature, gibberellic acid applications and paclobutrazol, an efficient plant growth retardant and fungicide at the same time. In this work, we have evaluated the effect of the auxin indole-3-acetic acid (IAA) dose (0, 1, 10, and 100 mg/plate) on the growth of the collection of B. cinerea isolates obtained from the following potted plants: Cyclamen persicum, Hydrangea macrophylla, Lantona camara, and Lonicera japonica. B. cinerea produces indolacetic acid, but so far the precise biosynthetic pathway and some effects on this fungal species are still unclear, although recent studies have revealed an antifungal activity of IAA on several fungi, including B. cinerea isolated from harvested fruits. Mycelial growth curves and growth rates assessed from difference in colony areas during the both linear and deceleration phase, conidiation (measured as time of appearance), conidia length (microm), and sclerotia production (number/plate) were evaluated in the isolates, which were grown at 26 degrees C on Petri dishes containing potato dextrose agar for up to 35 days. Mycelial growth curves fitted a typical kinetic equation of fungi grown on solid media. B. cinerea isolates showed a high degree of variability in their growth kinetics, depending on the isolate and auxin dose. This plant growth substance delayed mycelial growth during the linear phase in an isolate-dependent manner, thus isolates from C. persicum, H. macrophylla and L. camara were more affected by IAA than L. japonica. On the other hand, 100 mg of IAA was the critical dose to significantly reduce the growth rate in all isolates and to promote brown-striped hyphae development, especially in isolate from C. persicum. 10 and 100 mg

  19. Polyphasic characterization of Gluconacetobacter diazotrophicus isolates obtained from different sugarcane varieties.

    Science.gov (United States)

    Guedes, Helma V; Dos Santos, Samuel T; Perin, Liamara; Teixeira, Kátia R Dos S; Reis, Veronica M; Baldani, José I

    2008-10-01

    A polyphasic approach was applied to characterize 35 G. diazotrophicus isolates obtained from sugarcane varieties cultivated in Brazil. The isolates were analyzed by phenotypic (use of different carbon sources) and genotypic tests (ARDRA and RISA-RFLP techniques). Variability among the isolates was observed in relation to the carbon source use preference. Glucose and sucrose were used by all isolates in contrast to myo-inositol, galactose and ribose that were not metabolized. The results of the analysis showed the presence of two groups clustered at 68% of similarity. The genetic distance was higher when RISA-RFLP analysis was used. Analysis of 16S rDNA sequences from isolates showed that all of them belonged to the G. diazotrophicus species. Neither effect of the plant part nor sugarcane variety was observed during the cluster analysis. The observed metabolic and genetic variability will be helpful during the strain selection studies for sugarcane inoculation in association with sugarcane breeding programs.

  20. Transcriptome analysis of silkworm, Bombyx mori, during early response to Beauveria bassiana challenges.

    Science.gov (United States)

    Hou, Chengxiang; Qin, Guangxing; Liu, Ting; Geng, Tao; Gao, Kun; Pan, Zhonghua; Qian, Heying; Guo, Xijie

    2014-01-01

    Host-pathogen interactions are complex processes and it is a central challenge to reveal these interactions. Fungal infection of silkworm, Bombyx mori, may induce a variety of responsive reaction. However, little is known about the molecular mechanism of silkworm immune response against the fungal infection. To obtain an overview of the interaction between silkworm and an entomopathogenic fungus Beauveria bassiana, Digital Gene Expression profiling, a tag based high-throughput transcriptome sequencing method, was employed to screen and identify differentially expressed genes (DEGs, FDR ≤ 0.001, ∣log2ratio∣ ≥ 1) of silkworm larvae during early response against B. bassiana infection. Total 1430 DEGs including 960 up-regulated and 470 down-regulated ones were identified, of which 627 DEGs can be classified into GO categories by Gene Ontology (GO) analysis. KEGG pathways analysis of these DEGs suggested that many biological processes, such as defense and response, signal transduction, phagocytosis, regulation of gene expression, RNA splicing, biosynthesis and metabolism, protein transport etc. were involved in the interaction between the silkworm and B. bassiana. A number of differentially expressed fungal genes were also identified by mapping the sequencing tags to B. bassiana genome. These results provided new insights to the molecular mechanism of silkworm immune response to B. bassiana infection.

  1. The hidden habit of the entomopathogenic fungus Beauveria bassiana: first demonstration of vertical plant transmission.

    Directory of Open Access Journals (Sweden)

    Enrique Quesada-Moraga

    Full Text Available Beauveria bassiana strain 04/01-Tip, obtained from a larva of the opium poppy stem gall wasp Iraella luteipes (Hymenoptera; Cynipidae, endophytically colonizes opium poppy (Papaver somniferum L. plants and protects them against this pest. The goal of this study was to monitor the dynamics of endophytic colonization of opium poppy by B. bassiana after the fungus was applied to the seed and to ascertain whether the fungus is transmitted vertically via seeds. Using a species-specific nested PCR protocol and DNA extracted from surface-sterilised leaf pieces or seeds of B. bassiana-inoculated opium poppy plants, the fungus was detected within the plant beginning at the growth stage of rosette building and them throughout the entire plant growth cycle (about 120-140 days after sowing. The fungus was also detected in seeds from 50% of the capsules sampled. Seeds that showed positive amplification for B. bassiana were planted in sterile soil and the endophyte was again detected in more than 42% of the plants sampled during all plant growth stages. Beauveria bassiana was transmitted to seeds in 25% of the plants from the second generation that formed a mature capsule. These results demonstrate for the first time the vertical transmission of an entomopathogenic fungus from endophytically colonised maternal plants. This information is crucial to better understand the ecological role of entomopathogenic fungi as plant endophytes and may allow development of a sustainable and cost effective strategy for I. luteipes management in P. somniferum.

  2. The hidden habit of the entomopathogenic fungus Beauveria bassiana: first demonstration of vertical plant transmission.

    Science.gov (United States)

    Quesada-Moraga, Enrique; López-Díaz, Cristina; Landa, Blanca Beatriz

    2014-01-01

    Beauveria bassiana strain 04/01-Tip, obtained from a larva of the opium poppy stem gall wasp Iraella luteipes (Hymenoptera; Cynipidae), endophytically colonizes opium poppy (Papaver somniferum L.) plants and protects them against this pest. The goal of this study was to monitor the dynamics of endophytic colonization of opium poppy by B. bassiana after the fungus was applied to the seed and to ascertain whether the fungus is transmitted vertically via seeds. Using a species-specific nested PCR protocol and DNA extracted from surface-sterilised leaf pieces or seeds of B. bassiana-inoculated opium poppy plants, the fungus was detected within the plant beginning at the growth stage of rosette building and them throughout the entire plant growth cycle (about 120-140 days after sowing). The fungus was also detected in seeds from 50% of the capsules sampled. Seeds that showed positive amplification for B. bassiana were planted in sterile soil and the endophyte was again detected in more than 42% of the plants sampled during all plant growth stages. Beauveria bassiana was transmitted to seeds in 25% of the plants from the second generation that formed a mature capsule. These results demonstrate for the first time the vertical transmission of an entomopathogenic fungus from endophytically colonised maternal plants. This information is crucial to better understand the ecological role of entomopathogenic fungi as plant endophytes and may allow development of a sustainable and cost effective strategy for I. luteipes management in P. somniferum.

  3. OBTAINING THE POLAR COD PROTEIN ISOLATE AND ITS USING FOR MAYONNAISE AND MEATAND FISH FRANKFURTERS MANUFACTURING

    Directory of Open Access Journals (Sweden)

    V. I. Volchenko

    2015-01-01

    Full Text Available The technology of fish protein isolate from the low-cost raw material – the polar cod meat – was developed. The rationality of using partial alkaline-acid hydrolysis of minced fish for making fish protein isolate from the polar cod meat increasing the full-grade highly functional protein yield was proved. The most significant technological parameters influencing the physical and chemical characteristics of isolate were obtained; they are the parameters of washing and hydrolysis processes. The experiments of determination of the dependency of shelf life of the frozen raw material on physical and chemical characteristics of the raw material were carried out. The optimal quantity of washing cycles depending on the storage time was determined, it is 4 for the polar cod stored less than 4 month, and 6 for the polar cod stored more than 4 months. The dependency of temperature and pH of suspension changes during hydrolysis on the most significant quality characteristics (protein content, isolate yield by minced fish, isolate soluabilitywas obtained. The optimal values of these factors are: the temperature of 98 ˚С, pH of 11.75. The technology of manufacturing cholesterol-free mayonnaise choosing the fish protein isolate as an emulsifier was developed. The high emulsifying ability of fish protein isolate in fine emulsion “oil in water” was proved: increasing the content of isolate by 0.6 % the emulsifying ability increases by 1.85 %. The organ, oleptic microbiological, physical and chemical analyses of the mayonnaise were carried out, the recommended shelf life (30 days at the temperature from 3 to 5 ˚С was found. The fish protein isolate was also used for meat and fish frankfurters. The receipt of these frankfurters includes pork, FPI, microwave-blanched cod liver, salt, species, sugar, tomato paste, potato sctarch and yolk, liquid smoke.

  4. Allergens of the entomopathogenic fungus Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    Keyhani Nemat O

    2005-01-01

    Full Text Available Abstract Background Beauveria bassiana is an important entomopathogenic fungus currently under development as a bio-control agent for a variety of insect pests. Although reported to be non-toxic to vertebrates, the potential allergenicity of Beauveria species has not been widely studied. Methods IgE-reactivity studies were performed using sera from patients displaying mould hypersensitivity by immunoblot and immunoblot inhibition. Skin reactivity to B. bassiana extracts was measured using intradermal skin testing. Results Immunoblots of fungal extracts with pooled as well as individual sera showed a distribution of IgE reactive proteins present in B. bassiana crude extracts. Proteinase K digestion of extracts resulted in loss of IgE reactive epitopes, whereas EndoH and PNGaseF (glycosidase treatments resulted in minor changes in IgE reactive banding patterns as determined by Western blots. Immunoblot inhibitions experiments showed complete loss of IgE-binding using self protein, and partial inhibition using extracts from common allergenic fungi including; Alternaria alternata, Aspergillus fumigatus, Cladosporium herbarum, Candida albicans, Epicoccum purpurascens, and Penicillium notatum. Several proteins including a strongly reactive band with an approximate molecular mass of 35 kDa was uninhibited by any of the tested extracts, and may represent B. bassiana specific allergens. Intradermal skin testing confirmed the in vitro results, demonstrating allergenic reactions in a number of individuals, including those who have had occupational exposure to B. bassiana. Conclusions Beauveria bassiana possesses numerous IgE reactive proteins, some of which are cross-reactive among allergens from other fungi. A strongly reactive potential B. bassiana specific allergen (35 kDa was identified. Intradermal skin testing confirmed the allergenic potential of B. bassiana.

  5. Molecular and immunological characterization of allergens from the entomopathogenic fungus Beauveria bassiana

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    Keyhani Nemat O

    2006-09-01

    Full Text Available Abstract Background Entomopathogenic fungi such as Beauveria bassiana are considered promising biological control agents for a variety of arthropod pests. Beauveria species, however, have the potential to elicit allergenic reactions in humans, although no specific allergens have been characterized to date. Methods Four putative allergens were identified within B. bassiana expressed sequence tag (EST datasets. IgE-reactivity studies were performed using sera from patients displaying mold allergies against recombinant B. bassiana proteins expressed in E. coli. Results Full length cDNA and genomic nucleotide sequences of four potential B. bassiana allergens were isolated. BLASTX search results led to their putative designation as follows; Bb-Eno1, with similarity to fungal enolases; Bb-f2, similar to the Aspergillus fumigatus major allergen, Asp f2 and to a fibrinogen binding mannoprotein; Bb-Ald, similar to aldehyde dehydrogenases; and Bb-Hex, similar to N-acetyl-hexosaminadases. All four genes were cloned into E. coli expression systems and recombinant proteins were produced. Immunoblots of E. coli extracts probed with pooled as well as individual human sera from patients displaying mould allergies demonstrated IgE reactivity versus recombinant Bb-Eno1 and Bb-Ald. Conclusion Four putative Beauveria bassiana allergens were identified. Recombinant proteins corresponding to two of the four, Bb-Eno1 and Bb-Ald were bound by sera IgEs derived from patients with fungal allergies. These data confirm the potential allergenicity of B. bassiana by identification of specific human IgE reactive epitopes.

  6. Analysis of the genetic diversity of Candida isolates obtained from diabetic patients and kidney transplant recipients

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    Volmir Pitt Benedetti

    2016-01-01

    Full Text Available Yeasts of the genus Candida have high genetic variability and are the most common opportunistic pathogenic fungi in humans. In this study, we evaluated the genetic diversity among 120 isolates of Candida spp. obtained from diabetic patients, kidney transplant recipients and patients without any immune deficiencies from Paraná state, Brazil. The analysis was performed using the ITS1-5.8S-ITS2 region and a partial sequence of 28S rDNA. In the phylogenetic analysis, we observed a consistent separation of the species C. albicans, C. dubliniensis, C. glabrata, C. tropicalis, C. parapsilosis, C. metapsilosis and C. orthopsilosis, however with low intraspecific variability. In the analysis of the C. albicans species, two clades were formed. Clade A included the largest number of isolates (91.2% and the majority of isolates from GenBank (71.4%. The phylogenetic analysis showed low intraspecific genetic diversity, and the genetic polymorphisms between C. albicans isolates were similar to genetic divergence found in other studies performed with isolates from Brazil. This low genetic diversity of isolates can be explained by the geographic proximity of the patients evaluated. It was observed that yeast colonisation was highest in renal transplant recipients and diabetic patients and that C. albicans was the species most frequently isolated.

  7. Analysis of the genetic diversity of Candida isolates obtained from diabetic patients and kidney transplant recipients

    Science.gov (United States)

    Benedetti, Volmir Pitt; Savi, Daiani Cristina; Aluizio, Rodrigo; Adamoski, Douglas; Kava-Cordeiro, Vanessa; Galli-Terasawa, Lygia V; Glienke, Chirlei

    2016-01-01

    Yeasts of the genus Candida have high genetic variability and are the most common opportunistic pathogenic fungi in humans. In this study, we evaluated the genetic diversity among 120 isolates of Candida spp. obtained from diabetic patients, kidney transplant recipients and patients without any immune deficiencies from Paraná state, Brazil. The analysis was performed using the ITS1-5.8S-ITS2 region and a partial sequence of 28S rDNA. In the phylogenetic analysis, we observed a consistent separation of the species C. albicans, C. dubliniensis, C. glabrata, C. tropicalis, C. parapsilosis, C. metapsilosis and C. orthopsilosis, however with low intraspecific variability. In the analysis of the C. albicans species, two clades were formed. Clade A included the largest number of isolates (91.2%) and the majority of isolates from GenBank (71.4%). The phylogenetic analysis showed low intraspecific genetic diversity, and the genetic polymorphisms between C. albicans isolates were similar to genetic divergence found in other studies performed with isolates from Brazil. This low genetic diversity of isolates can be explained by the geographic proximity of the patients evaluated. It was observed that yeast colonisation was highest in renal transplant recipients and diabetic patients and that C. albicans was the species most frequently isolated. PMID:27276363

  8. Studies on the Virome of the Entomopathogenic Fungus Beauveria bassiana Reveal Novel dsRNA Elements and Mild Hypervirulence.

    Science.gov (United States)

    Kotta-Loizou, Ioly; Coutts, Robert H A

    2017-01-01

    The entomopathogenic fungus Beauveria bassiana has a wide host range and is used as a biocontrol agent against arthropod pests. Mycoviruses have been described in phytopathogenic fungi while in entomopathogenic fungi their presence has been reported only rarely. Here we show that 21.3% of a collection of B. bassiana isolates sourced from worldwide locations, harbor dsRNA elements. Molecular characterization of these elements revealed the prevalence of mycoviruses belonging to the Partitiviridae and Totiviridae families, the smallest reported virus to date, belonging to the family Narnaviridae, and viruses unassigned to a family or genus. Of particular importance is the discovery of members of a newly proposed family Polymycoviridae in B. bassiana. Polymycoviruses, previously designated as tetramycoviruses, consist of four non-conventionally encapsidated capped dsRNAs. The presence of additional non-homologous genomic segments in B. bassiana polymycoviruses and other fungi illustrates the unprecedented dynamic nature of the viral genome. Finally, a comparison of virus-free and virus-infected isogenic lines derived from an exemplar B. bassiana isolate revealed a mild hypervirulent effect of mycoviruses on the growth of their host isolate and on its pathogenicity against the greater wax moth Galleria mellonella, highlighting for the first time the potential of mycoviruses as enhancers of biocontrol agents.

  9. Production of extracellular proteolytic enzymes by Beauveria bassiana

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    Józefa Chrzanowska

    2014-08-01

    Full Text Available The production of proteolytic enzymes by two strains of Beauveria bassiana 278, B. bassiana 446 and one strain of Ascosphera apis 496 was analysed. It was demonstrated that the strain of B. bassiana 278 proved to be the best producer of basic and acid proteases. The influence of different environmental factors such as nitrogen and carbon sources on the production of extracellular hydrolytic enzymes was assessed. In addition the acid protease from B. bassiana was partially characterized.

  10. Production of extracellular proteolytic enzymes by Beauveria bassiana

    OpenAIRE

    Józefa Chrzanowska; Maria Kołaczkowska

    2014-01-01

    The production of proteolytic enzymes by two strains of Beauveria bassiana 278, B. bassiana 446 and one strain of Ascosphera apis 496 was analysed. It was demonstrated that the strain of B. bassiana 278 proved to be the best producer of basic and acid proteases. The influence of different environmental factors such as nitrogen and carbon sources on the production of extracellular hydrolytic enzymes was assessed. In addition the acid protease from B. bassiana was partially characterized.

  11. Evaluation of three formulations of Beauveria bassiana for control of lesser mealworm and hide beetle in Georgia poultry houses

    Science.gov (United States)

    Initial screening of 12 Beauveria bassiana (Balsamo) Vuillemin isolates against larvae of the lesser mealworm (Alphitobius diaperinus[Panzer]) resulted in the selection of two isolates, GHA and 707, for further testing under field conditions. Three formulations of each strain were prepared: an EC, ...

  12. Pathogenicity and phenotypic sulfadiazine resistance of Toxoplasma gondii isolates obtained from livestock in northeastern Brazil.

    Science.gov (United States)

    Oliveira, Claudio Bs; Meurer, Ywlliane Sr; Andrade, Joelma Ma; Costa, Maria Esm; Andrade, Milena Mc; Silva, Letícia A; Lanza, Daniel Cf; Vítor, Ricardo Wa; Andrade-Neto, Valter F

    2016-06-01

    Toxoplasma gondii is the causative protozoan agent of toxoplasmosis, which is a common infection that is widely distributed worldwide. Studies revealed stronger clonal strains in North America and Europe and genetic diversity in South American strains. Our study aimed to differentiate the pathogenicity and sulfadiazine resistance of three T. gondii isolates obtained from livestock intended for human consumption. The cytopathic effects of the T. gondii isolates were evaluated. The pathogenicity was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using a CS3 marker and in a rodent model in vivo. Phenotypic sulfadiazine resistance was measured using a kinetic curve of drug activity in Swiss mice. IgM and IgG were measured by ELISA, and the dihydropteroate synthase (DHPS) gene sequence was analysed. The cytopathic effects and the PCR-RFLP profiles from chickens indicated a different infection source. The Ck3 isolate displayed more cytopathic effects in vitro than the Ck2 and ME49 strains. Additionally, the Ck2 isolate induced a differential humoral immune response compared to ME49. The Ck3 and Pg1 isolates, but not the Ck2 isolate, showed sulfadiazine resistance in the sensitivity assay. We did not find any DHPS gene polymorphisms in the mouse samples. These atypical pathogenicity and sulfadiazine resistance profiles were not previously reported and served as a warning to local health authorities.

  13. Experimental infections with Paracoccidioides brasiliensis obtained from armadillos: comparison to clinical isolates

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    Angela Satie Nishikaku

    2008-02-01

    Full Text Available Paracoccidioides brasiliensis causes paracoccidioidomycosis (PCM that is one of the most prevalent systemic human mycoses in Latin America. Armadillos show a high incidence of PCM infection and could, therefore, be a natural reservoir for this fungus. In this study were compared the virulence profiles of isolates obtained from nine-banded armadillos (Dasypus novemcinctus (PbT1 and PbT4 and isolates from PCM patients (Pb265 and Bt83. Pathogenicity was evaluated by fungal load and analysis of colony morphology. Immunity against the fungus was tested by delayed type hypersensitivity test (DTH and antibody quantification by ELISA. The higher virulence of PbT1 and PbT4 was suggested by higher fungal load in spleen and lungs. Armadillo isolates and Bt83 presented a cotton-like surface contrasting with the cerebriform appearance of Pb265. All isolates induced cellular and humoral immune responses in infected BALB/c mice. DTH reactions were similarly induced by the four isolates, however, a great variability was observed in specific antibody levels, being the highest ones induced by Bt83 and PbT4. The present work confirms that armadillos harbor P. brasiliensis, whose multiplication and induced immunity in experimentally infected mice are heterogeneous, resembling the behavior of isolates from human PCM. This study reinforces the possibility that armadillos play an important role in the biological cycle of this pathogen.

  14. Identification of virulence genes carried by bacteriophages obtained from clinically isolated methicillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Karasartova, Djursun; Cavusoglu, Zeynep Burcin; Turegun, Buse; Ozsan, Murat T; Şahin, Fikret

    2016-12-01

    Bacteriophages play an important role in the pathogenicity of Staphylococcus aureus (S. aureus) either by carrying accessory virulence factors or several superantigens. Despite their importance, there are not many studies showing the actual distribution of the virulence genes carried by the prophages obtained from the clinically isolated Staphylococcus. In this study, we investigated prophages obtained from methicillin-resistant S. aureus (MRSA) strains isolated from hospital- and community-associated (HA-CA) infections for the virulence factors. In the study, 43 phages isolated from 48 MRSA were investigated for carrying toxin genes including the sak, eta, lukF-PV, sea, selp, sek, seg, seq chp, and scn virulence genes using polymerase chain reaction (PCR) and Southern blot. Restriction fragment length polymorphism was used to analyze phage genomes to investigate the relationship between the phage profiles and the toxin genes' presence. MRSA strains isolated from HA infections tended to have higher prophage presence than the MRSA strains obtained from the CA infections (97% and 67%, respectively). The study showed that all the phages with the exception of one phage contained one or more virulence genes in their genomes with different combinations. The most common toxin genes found were sea (83%) followed by sek (77%) and seq (64%). The study indicates that prophages encode a significant proportion of MRSA virulence factors.

  15. Copaifera langsdorffii: evaluation of potential gastroprotective of extract and isolated compounds obtained from leaves

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    Marivane Lemos

    2015-06-01

    Full Text Available AbstractGastric ulcer is a prevalent gastrointestinal disease, and the drugs currently used in the treatment produce several adverse effects. In this context, the search for new therapeutic antiulcer agents is essential, and medicinal plants have great potential. Here, we investigated the gastroprotective properties of Copaifera langsdorffii Desf., Fabaceae, hydroalcoholic extract obtained from leaves and its isolated compounds. The phytochemistry studies and the compounds isolations were performed using chromatographic and spectroscopic methodologies. The hydroalcoholic extract was evaluated using ethanol/HCl, non-steroidal anti-inflammatory drug, stress-induced-ulcer and chronic ulcer-model. The effects on gastric content volume, pH, total acidity and mucus stomach production were evaluated in the pylorus ligated-model. The C. langsdorffii extract obtained from leaves (50, 250 or 500 mg/kg reduced the injured area compared to control group in all experiments. The extract showed a significant decrease in the total gastric juice acidity and an increase in mucus production (500 mg/kg when compared to vehicle. Among isolated compounds (30 mg/kg α-humulene, β-caryophyllene and caryophyllene oxide showed greater gastroprotective activity in the ethanol/HCl induced ulcer model. The data herein obtained shown that C. langsdorffii leaves extract and isolated compounds from it, presented gastroprotective properties in different animal models of gastric ulcer. These effects may be associated with the ability of the extract to decrease gastric secretion and increase the mucus production.

  16. Characterization of rhizobia isolates obtained from nodules of wild genotypes of common bean.

    Science.gov (United States)

    Cardoso, Aline Assis; Andraus, Michel de Paula; Borba, Tereza Cristina de Oliveira; Martin-Didonet, Claudia Cristina Garcia; Ferreira, Enderson Petrônio de Brito

    This study aimed to evaluate the tolerance to salinity and temperature, the genetic diversity and the symbiotic efficiency of rhizobia isolates obtained from wild genotypes of common bean cultivated in soil samples from the States of Goiás, Minas Gerais and Paraná. The isolates were subjected to different NaCl concentrations (0%, 1%, 2%, 4% and 6%) at different temperatures (28°C, 33°C, 38°C, 43°C and 48°C). Genotypic characterization was performed based on BOX-PCR, REP-PCR markers and 16S rRNA sequencing. An evaluation of symbiotic efficiency was carried out under greenhouse conditions in autoclaved Leonard jars. Among 98 isolates about 45% of them and Rhizobium freirei PRF81 showed a high tolerance to temperature, while 24 isolates and Rhizobium tropici CIAT899 were able to use all of the carbon sources studied. Clustering analysis based on the ability to use carbon sources and on the tolerance to salinity and temperature grouped 49 isolates, R. tropici CIAT899 and R. tropici H12 with a similarity level of 76%. Based on genotypic characterization, 65% of the isolates showed an approximately 66% similarity with R. tropici CIAT899 and R. tropici H12. About 20% of the isolates showed symbiotic efficiency similar to or better than the best Rhizobium reference strain (R. tropici CIAT899). Phylogenetic analysis of the 16S rRNA revealed that two efficient isolates (ALSG5A1 and JPrG6A8) belong to the group of strains used as commercial inoculant for common bean in Brazil and must be assayed in field experiments.

  17. Relationship between Biological Characteristics of Beauveria bassiana (Bals.) Vuill and Pathogenicity to Bombyx mori L.%Relationship between Biological Characteristics of Beauveria bassiana (Bals.) Vuill and Pathogenicity to Bombyx mori L.

    Institute of Scientific and Technical Information of China (English)

    Haiyu LUO; Yecheng DENG; Yongmei LIAO; Ruiyu LI

    2012-01-01

    [Objective] This study was to investigate the relationship between biological characteristics of Beauveria bassiana (Bals.) Vuill and pathogenicity to Bombyx rnori L, with the aim to provide scientific basis for the control of white muscardine in Bombyx mori L. [Method] The strains isolated and purified from the 6 Beauveria bassiana biocontrol agents from all over the country and the 3 white muscardine silkworms collected from Guangxi provincial silkworm rearing areas were identified by the morphological observation and molecular biology technology. The pathogenicity of B. bassaina to silkworms was determined, and the biological characteristics such as growth diameter, sporulation and the extracellular protease activity of the different B. bassiana strains were compared. [Result] The isolated 9 strains were all B. bassaina (Bals.) Vuillemin, and all strains had high pathogenicity to silkworm, but with different pathogenicities. The growth diameter, sporulation and extracellular protease activity of different B. bassiana strains were also different, and showed correlation with the patheogenicity to silkworms. [Conclusion] B. bassiana spores production amount and exocellular protease activity had significant positive correlation with their pathogenicity to silkworm.

  18. Pathogenicity and phenotypic sulfadiazine resistance ofToxoplasma gondii isolates obtained from livestock in northeastern Brazil

    Science.gov (United States)

    Oliveira, Claudio BS; Meurer, Ywlliane SR; Andrade, Joelma MA; Costa, Maria ESM; Andrade, Milena MC; Silva, Letícia A; Lanza, Daniel CF; Vítor, Ricardo WA; Andrade-Neto, Valter F

    2016-01-01

    Toxoplasma gondii is the causative protozoan agent of toxoplasmosis, which is a common infection that is widely distributed worldwide. Studies revealed stronger clonal strains in North America and Europe and genetic diversity in South American strains. Our study aimed to differentiate the pathogenicity and sulfadiazine resistance of three T. gondiiisolates obtained from livestock intended for human consumption. The cytopathic effects of the T. gondii isolates were evaluated. The pathogenicity was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using a CS3 marker and in a rodent model in vivo. Phenotypic sulfadiazine resistance was measured using a kinetic curve of drug activity in Swiss mice. IgM and IgG were measured by ELISA, and the dihydropteroate synthase (DHPS) gene sequence was analysed. The cytopathic effects and the PCR-RFLP profiles from chickens indicated a different infection source. The Ck3 isolate displayed more cytopathic effects in vitro than the Ck2 and ME49 strains. Additionally, the Ck2 isolate induced a differential humoral immune response compared to ME49. The Ck3 and Pg1 isolates, but not the Ck2 isolate, showed sulfadiazine resistance in the sensitivity assay. We did not find any DHPS gene polymorphisms in the mouse samples. These atypical pathogenicity and sulfadiazine resistance profiles were not previously reported and served as a warning to local health authorities. PMID:27276184

  19. Isolation and identification of acetogenic bacteria obtained from deer rumen and their potential for methanogenesis inhibitor

    Directory of Open Access Journals (Sweden)

    Amlius Thalib

    2008-10-01

    Full Text Available Methanogenesis can be inhibited by various chemicals through different mechanism reaktion. The use of acetogenic bacteria as H2 sink is assumed to be a promising approach. Isolation and identification of acetogenic bacteria obtained from deer rumen had been conducted. Two types of media used for isolation were hydrogen-carbondioxide utilizing acetogens and carbonmonoxide utilizing acetogens. Identification of species of acetogens isolates was based on descriptions of morphology, Gram type, motility, bioreaction results, and oksygen requirement. The compositions of methane and volatile fatty acids (VFA were determined on minimal media or added with sheep rumen liquid innoculated with pure isolates. The identification results showed that the isolate cultured on media of hydrogen-carbondioxide utilizing acetogens was Acetoanaerobium noterae and the ones cultured on media of carbonmonoxide utilizing acetogens was Acetobacterium woodii. Inoculumn of A. noterae and A. woodii could decreased the composition of methane resulted from substrate fermented by fresh rumen liquid of sheep (CRDF, that is culture of A. noterae added FPM and defaunator decreased methane production by 28.8% (P CH3COOH + 2H2O by which reduction of CO2 with H2 producing CH4 can be inhibited or decreased. Their function as methanogenesis inhibitor would be more significant when they are combined with microbial growth factors and defaunator.

  20. SEROLOGICAL DETECTION AND MOLECULAR CHARACTERIZATION OF A BEGOMOVIRUS ISOLATE OBTAINED FROM Macroptilium lathyroides

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    JOSÉ ALBERSIO DE ARAUJO LIMA

    2012-01-01

    Full Text Available The viruses from the genus Begomovirus, family Geminiviridae are considered emergent pathogens, mainly because of the population explosion of their insect vectors. For this reason, more attention needs to be directed to the correct virus species identification inside the genus. The present paper had the objectives of serologically detecting a begomovirus in Macroptilium lathyroides plants in the State of Ceará, and developing biological, serological and molecular studies with a virus isolate obtained from M. lathyroides. Indirect ELISA with antiserum for Macroptilium golden mosaic virus (MaGMV demonstrated that the samples collected from M. lathyroides showing golden mosaic in the field were infected with a begomovirus. The virus isolate obtained was transmitted by grafting to eight species of the family Leguminosae, four species of Solonaceae, and one species in the family Amaranthaceae. The virus also was transmitted from M. lathyroides to M. lathyroides by the whitefly Bemisia tabaci biotype B. A DNA fragment of 1.2 kb was obtained by PCR with the primers PAL1v 1978 and PAR1c 496 for component A, and a DNA fragment of 0.5 kb was obtained with the primers PBL1v 2040 and PCR cl for component B, confirming the presence of a begomovirus infecting M. lathyroides. Molecular studies indicated that the begomovirus isolate showed 77% genomic similarity with Bean golden mosaic virus and 75% with Cowpea golden mosaic virus for their cp and rep genes, indicating the possibility that the isolate is a distinct virus species of the Begomovirus genus.

  1. Effect of surfactants and temperature on germination and vegetative growth of Beauveria bassiana

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    Lizzy A. Mwamburi

    2015-03-01

    Full Text Available Three non-ionic surfactants: Tween20, Tween80 and Breakthru® were screened for their effects on spore germination and mycelial growth rates and for their influence on three isolates of Beauveria bassianaspore germination at various temperatures. Tween20 and Tween80 were compatible with all the B. bassiana isolates in the germination studies, but inhibited germination at higher surfactant concentrations, irrespective of the conidial concentrations. Breakthru® had an inhibitory effect on germination even at the lowest concentration of 0.1% on all the B. bassiana isolates. The effects of the surfactants on spore germination did not correspond with their effects on colony growth. Conidial viability within the same formulation declined significantly with increases in temperature, irrespective of the surfactant. The optimal temperature for conidial germination of B. bassiana isolates was approximately 25 °C with an upper limit at 30 °C. Isolate 7320 was identified as the least affected by the different surfactants. This isolate was able to germinate rapidly in a broad temperature range of 25–30 °C after 24 h, this characteristic being an essential factor in controlling house fly populations in poultry houses.

  2. Effect of Leaf Litters and Soils on Viability of Entomopathogenic Fungi Beauveria bassiana (Bals. Vuill

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    LISDAR IDWAN SUDIRMAN

    2008-09-01

    Full Text Available Viability of Beauveria bassiana is extremely low due to toxic compounds in soils. This research was aimed to study the effect of four groups of media on viability of B. bassiana Bb-Pb2. The first group was leaf litters of onion, flowering white cabbage, cabbage, and chinese mustard, respectively; the second group was the soils containing decomposed residues of each plant of the first group; the third group was the mixtures of each media of both groups above (1:1, and the fourth group was natural top soil as a control. Each plastic bag filled with one kg of each medium was inoculated with ten ml of B. bassiana conidia (106/ml of concentration and incubated in open area for 8 weeks. The results showed that all leaf litters of those plants and their compost soils affected the fungal viability. The highest decreasing number of colony was found on onion’s leaf litters, soil containing of decomposed onion, and the mixtures of both media. The treated B. bassiana showed significant reducing abilities of growth, conidia production and conidia germination on PDA media, except the one of control. It is suggested that the Bb-Pb2 isolate might not be effective as bioinsecticide in the soils containing either those leaf litters or composts.

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

    Science.gov (United States)

    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.

  4. A novel isolation strategy for obtaining crude membrane vesicles from bovine skim milk

    DEFF Research Database (Denmark)

    Blans, Kristine; Larsen, Lotte Bach; Wiking, Lars

    2014-01-01

    Bovine milks content of phospholipid membranes have largely been explored in the cream fraction, and known as the milk fat globule membrane that surrounds fat droplets. In skim milk, the population of phospholipid membranes is reported to constitute membrane vesicles with a soluble content known...... components. Here we present a novel strategy for a short, gentle and non-denaturing isolation of skim milk-derived membrane vesicles. Methods: Untreated fresh bovine milk was defatted to remove milk fat globules. The resulting skim milk was subjected to ultracentrifugation. The resulting ochre...... fraction can be obtained from skim milk by ultracentrifugation. Casein micelle remnants as well as smaller protein components in the crude vesicle fraction can be successfully removed by size chromatography. Electron microscopy of the vesicle isolate reveals circular structures with membrane vesicle...

  5. A novel isolation strategy for obtaining crude membrane vesicles from bovine skim milk

    DEFF Research Database (Denmark)

    Blans, Kristine; Larsen, Lotte Bach; Wiking, Lars

    Bovine milks content of phospholipid membranes have largely been explored in the cream fraction, and known as the milk fat globule membrane that surrounds fat droplets. In skim milk, the population of phospholipid membranes is reported to constitute membrane vesicles with a soluble content known...... components. Here we present a novel strategy for a short, gentle and non-denaturing isolation of skim milk-derived membrane vesicles. Methods: Untreated fresh bovine milk was defatted to remove milk fat globules. The resulting skim milk was subjected to ultracentrifugation. The resulting ochre...... fraction can be obtained from skim milk by ultracentrifugation. Casein micelle remnants as well as smaller protein components in the crude vesicle fraction can be successfully removed by size chromatography. Electron microscopy of the vesicle isolate reveals circular structures with membrane vesicle...

  6. 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.

  7. Soil application of Beauveria bassiana GHA against apple sawfly, Hoplocampa testudinea (Hymenoptera: Tenthredinidae): Field mortality and fungal persistence.

    Science.gov (United States)

    Świergiel, Weronika; Meyling, Nicolai V; Porcel, Mario; Rämert, Birgitta

    2016-12-01

    Low impact alternatives to synthetic insecticides for the control of apple sawfly (Hoplocampa testudinea Klug) are scarce encumbering pest management in organic apple orchards. We investigated the soil persistence and field efficacy of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (BotaniGard) against apple sawfly under common organic orchard practices. We also assessed the efficacy of B. bassiana GHA and Metarhizium brunneum Petch (indigenous strain) against sawfly in the laboratory. Larvae treated with either fungus in the laboratory died faster than control larvae and displayed 49.4%-68.4% mycosis. In the field, B. bassiana density remained high in the week after application, during larval descent to the soil. Fungal density decreased to 25% at 49 d after application and to 0.4% after 55 weeks. Molecular markers revealed that the majority of fungal isolates recovered comprised the applied B. bassiana strain GHA. Larvae pupating in soil cages in the orchard for 49 d displayed 17% mycosis. The high efficacy under laboratory conditions was not seen in the field. B. bassiana application resulted in densities above the upper natural background level during the growing season, but reversion to background levels occurred within a year. It remains to be investigated whether this has a detrimental effect on nontarget organisms. Additional work is needed to bridge the knowledge gap between laboratory and field efficacy in orchards.

  8. Genotyping of methicillin-resistant Staphylococcus aureus isolates obtained in the Northeast region of Brazil

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    F.C. de Sousa-Junior

    2009-10-01

    Full Text Available Methicillin-resistant Staphylococcus aureus (MRSA is a major agent of hospital infections worldwide. In Brazil, a multiresistant MRSA lineage (ST239-SCCmecIIIA, the so-called Brazilian epidemic clone (BEC, has predominated in all regions. However, an increase in nosocomial infections caused by non-multiresistant MRSA clones has recently been observed. In the present study, 45 clinical isolates of MRSA obtained from a university hospital located in Natal city, Brazil, were identified by standard laboratory methods and molecularly characterized using staphylococcal chromosome cassette mec (SCCmec typing and pulsed-field gel electrophoresis. Antimicrobial susceptibility testing was carried out using CLSI methods. The MRSA isolates studied displayed a total of 8 different pulsed-field gel electrophoresis patterns (types A to H with predominance (73% of pattern A (BEC-related. However, MRSA harboring SCCmec type IV were also identified, 3 (7% of which were genetically related to the pediatric clone - USA800 (ST5-SCCmecIV. In addition, we found a considerable genetic diversity within BEC isolates. MRSA displaying SCCmecIV are frequently susceptible to the majority of non-β-lactam antibiotics. However, emergence of multiresistant variants of USA800 was detected.

  9. Amylase production potentials of bacterial isolates obtained from the gut of Oryctes rhinoceros larvae

    Science.gov (United States)

    Aryati, P. C.; Pangastuti, A.; Sari, S. L. A.

    2017-04-01

    Amylase is one of the main enzymes used in industry, such as food, detergent, textile, and pharmaceutical industry. Amylase can be produced by plants, animals, and microorganisms. However, bacterial and fungal amylases have dominated application in industries. This research was aimed to determine amylolytic activity of bacteria isolated from the gut of Oryctes rhinoceros larvae. Based on clear zone formation, 9 from 11 isolates showed amylolytic activity. Isolates with the widest clear zone, i.e Bacillus subtilis GOR1, Bacillus cereus GOR3, and Bacillus pumilus GOR2, were screened for amylolytic activity based on reduction sugar production. The result showed that Bacillus subtilis GOR1 was the most potential as amylase producer, showed by the widest clear zone 5.224 cm2 and highest reduction sugar production 0.0235 mg/ml. Highest amylase specific activity (0.1447 U/mg protein) was obtained at 60°C and pH 7. Amylase activity was stable for 3 hours at 60°C with residual activity respectively was 59.7%.

  10. Nanocomposites films obtained from protein isolates of mechanically deboned chicken meat added with montmorillonite

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    Bruna da Silva Menezes

    Full Text Available Abstract The aim of this study was to evaluate the properties of nanocomposite films of protein isolates from mechanically deboned chicken meat with organoclay (montmorillonite. For the film development, a 23 experimental design was performed with three levels, protein isolate (2, 3.5, 5 g.100 mL-1 of solution, montmorillonite (0.3, 0.5, 0.7 g.100mL-1 of solution and glycerol (25, 30, 35 g.100 mL-1 CPI. The tensile strength varied between 6.7 and 9.1 MPa, elongation to break from 26-66%, opacity of 13.1 to 35.7 and solubility from 38.5% to 81.8%. Assessing the structural properties, interleaving of the isolate and montmorillonite can be noted. The results obtained in the experimental design indicate that 2.0 g of CPI.100 g-1 of solution, 0.8 g of MMT.100 g-1of solution and 0.2 g of glicerol.100 g-1CPI are the ideal parameters for preparing nanocomposite films.

  11. 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.

  12. 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

  13. Simple and efficient method for isolation and cultivation of endoscopically obtained human colonocytes

    DEFF Research Database (Denmark)

    Seidelin, Jakob B; Horn, Thomas; Nielsen, Ole H

    2003-01-01

    on a porous membrane was superior to the other methods applied [P cultivated cells. CM had similar viability supporting effects to FCS. Other supplements had......Few comparative and validated reports exist on the isolation and growth of colonoscopically obtained colonic epithelium. The aim of this study was to develop and validate a simple method for the cultivation of colonoscopically obtained colonocytes. Forty patients, who underwent routine colonoscopy...... and where the diagnosis of irritable bowel syndrome was later reached, were included. Seven colon biopsies were taken and incubated at varying time periods of 10-120 min and temperatures of 4-37 degrees C in a chelating buffer. The epithelium was then harvested and cultivated under three different...

  14. Simple and efficient method for isolation and cultivation of endoscopically obtained human colonocytes

    DEFF Research Database (Denmark)

    Seidelin, Jakob B; Horn, Thomas; Nielsen, Ole H

    2003-01-01

    on a porous membrane was superior to the other methods applied [P cultivated cells. CM had similar viability supporting effects to FCS. Other supplements had no effects......Few comparative and validated reports exist on the isolation and growth of colonoscopically obtained colonic epithelium. The aim of this study was to develop and validate a simple method for the cultivation of colonoscopically obtained colonocytes. Forty patients, who underwent routine colonoscopy...... and where the diagnosis of irritable bowel syndrome was later reached, were included. Seven colon biopsies were taken and incubated at varying time periods of 10-120 min and temperatures of 4-37 degrees C in a chelating buffer. The epithelium was then harvested and cultivated under three different...

  15. Isolation and Identification of Pathogenic Fungi-Beauveria bassiana%沙棘木蠹蛾幼虫病原真菌——球孢白僵菌的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    王素英; 特木钦; 王英凯; 陆群; 时亚琴; 杨奋勇; 苏梅

    2007-01-01

    在对沙棘木蠹蛾危害情况进行调查时,从死亡的沙棘树干内发现了罹病致死的沙棘木蠹蛾,通过对虫尸体内病原真菌的分离与鉴定,确定为球孢白僵菌(Beauveria bassiana).经室内致病力测定,接种浓度4亿/mL、8亿/mL、12亿/mL,54 d的感病率分别是72.22%、83.33%、100%,证明球孢白僵菌是沙棘木蠹蛾的病原菌.同时表明浓度越高感病率越高.

  16. Beauveria bassiana Strains for Biological Control of Cosmopolites sordidus (Germ. (Coleoptera: Curculionidae in Plantain

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    Marilene Fancelli

    2013-01-01

    Full Text Available The objective of this study was to select strains of Beauveria bassiana for controlling Cosmopolites sordidus (Germ. in plantain farms (cv. Terra of the “Recôncavo” and southern regions in the state of Bahia, Brazil. The virulence of 32 B. bassiana isolates against C. sordidus was determined under laboratory conditions. Three isolates (CNPMF 407, CNPMF 218, and CNPMF 416 were selected for evaluation under field conditions in plantations located in the counties of Mutuípe and Wenceslau Guimarães. Population of C. sordidus was estimated every 15 days by using pseudostem traps. The efficiency of the three strains of B. bassiana was compared to chemical control (carbofuran, 4g/trap and absence of control. Carbofuran caused around 90% of adult mortality after 12 months, with a reduction in the population of C. sordidus since the first evaluation. A low number of trapped insects was observed in the fungus-treated plots, suggesting the efficiency of the isolates in controlling the C. sordidus population. The strain CNPMF 218 was the most efficient in controlling C. sordidus adults in both locations, causing around 20% mortality, leading to 40% population size reduction after 12 months.

  17. Simple and efficient method for isolation and cultivation of endoscopically obtained human colonocytes.

    Science.gov (United States)

    Seidelin, Jakob B; Horn, Thomas; Nielsen, Ole H

    2003-12-01

    Few comparative and validated reports exist on the isolation and growth of colonoscopically obtained colonic epithelium. The aim of this study was to develop and validate a simple method for the cultivation of colonoscopically obtained colonocytes. Forty patients, who underwent routine colonoscopy and where the diagnosis of irritable bowel syndrome was later reached, were included. Seven colon biopsies were taken and incubated at varying time periods of 10-120 min and temperatures of 4-37 degrees C in a chelating buffer. The epithelium was then harvested and cultivated under three different conditions: 1) on a collagen coating, 2) embedded in a collagen gel, or 3) embedded in a gel put on a porous well insert. The effect of conditioned medium (CM), insulin, transferrin, selenium, and the oxygen content was assessed. Viability was tested by the metabolic dimethylthiazol-diphenyl-tetrazolium bromide assay, by flowcytometry, by phase contrast microscopy, and by transmission electron microscopy. Incubation at 21 degrees C for 75 min gave an optimal yield of 3 x 10(6) (2.0-3.8 x 10(6)) viable epithelial cells in intact crypts per seven biopsies. Embedding of crypts in a collagen gel put on a porous membrane was superior to the other methods applied [P coat-cultivated cells. CM had similar viability supporting effects to FCS. Other supplements had no effects. A simple method is presented, which makes cultivation of colonocytes obtained at endoscopy possible for up to 72 h.

  18. 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.

  19. 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)

    Energy Technology Data Exchange (ETDEWEB)

    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)

  20. Exploiting the genetic diversity of Beauveria bassiana for improving the biological control of the coffee berry borer through the use of strain mixtures.

    Science.gov (United States)

    Cruz, Lina P; Gaitan, Alvaro L; Gongora, Carmenza E

    2006-08-01

    Beauveria bassiana is an entomopathogen widely used to control the coffee berry borer in Colombia, as part of an Integrated Pest Management strategy. Traditionally, the development of fungal insect pathogens as biocontrol agents in crop pests has been oriented towards the selection and formulation of elite clonal strains. Instead, we explored the potential application of genetic diversity in B. bassiana by determining the effect of strain mixtures on coffee berry borer mortality compared to clonal isolates. Genomic DNA from 11 strains was characterized using internal transcribed spacers and beta-tubulin sequences as well as amplified fragment length polymorphism markers. Cluster analysis produced three genetic groups and confirmed the low but significant intraspecific genetic diversity present among the strains. Single strain virulence towards the coffee berry borer under laboratory conditions, using 1x10(6) conidia ml(-1), ranged between 89.9 and 57.5%. All the inoculations with mixtures resulted in coinfection events. Combinations of genetically similar strains showed no significant differences when their virulences were compared. However, mixtures of genetically different strains led to both antagonism and synergism. The lowest virulence percentage (57%) was obtained by putting together the most virulent strain of each group, contrary to the highest virulence percentage (93%) that resulted from mixing the three least virulent strains. The results indicate the promising potential of designing strain mixtures as an alternative for the biocontrol of Hypothenemus hampei and other pests and provide tools for the understanding of the ecological dynamics of entomopathogen populations under natural conditions.

  1. THE EFFECT OF SELECTED PESTICIDES ON THE GROWTH OF ENTOMOPATHOGENIC FUNGI HIRSUTELLA NODULOSA AND BEAUVERIA BASSIANA

    Directory of Open Access Journals (Sweden)

    Cezary Tkaczuk

    2015-06-01

    Full Text Available The effect of three pesticides used in corn protection: Targa Super 05 (chizalofop-P-ethyl, Roundup 360SL (glyphosate and Karate Zeon 050CS (lambda-cyhalothrin, on the growth of entomopathogenic fungi Hirsutella nodulosa Petch and Beauveria bassiana (Bals. Vuill. was evaluated under laboratory conditions. Fungi isolates were cultured on Sabouraud’s medium with addition of pesticides at three different concentrations. H. nodulosa was more susceptible to pesticides than B. bassiana. The most inhibitory effect on tested entomopathogenic fungi showed chizalofop-P-ethyl herbicide. Lambda-cyhalothrin at the recommended field dose showed minor toxic effect on entomopathogenic fungi, which suggests the possibility of compatible use of this insecticide with biopesticides based on the tested species.

  2. Biocide and antibiotic susceptibility of Salmonella isolates obtained before and after cleaning at six Danish pig slaughterhouses

    DEFF Research Database (Denmark)

    Gantzhorn, Mette Rørbæk; Pedersen, Karl; Olsen, John Elmerdahl

    2014-01-01

    . The susceptibility toward three different biocides, triclosan and two commercial disinfection products: Desinfect Maxi, a quaternary ammonium compound, and Incimaxx DES, an acetic compound, was determined. We found no resistance toward the biocides tested, but we did find that isolates obtained after cleaning had...... that there was a weak statistical correlation between MICs toward the biocides and some antibiotics, but no difference in log(MIC)s toward antibiotics between isolates obtained before and after cleaning, nor did we find any difference in the number of resistances of isolates obtained before and after cleaning...

  3. A technique for the prevention of greenhouse whitefly (Trialeurodes vaporariorum) using the entomopathogenic fungus Beauveria bassiana M130.

    Science.gov (United States)

    Kim, Chang-Su; Lee, Jung-Bok; Kim, Beam-Soo; Nam, Young-Ho; Shin, Kee-Sun; Kim, Jin-Won; Kim, Jang-Eok; Kwon, Gi-Seok

    2014-01-01

    The possibility of using hyphomycete fungi as suitable biocontrol agents against greenhouse whitefly has led to the isolation of various insect pathogenic fungi. Among them is Beauveria bassiana, one of the most studied entomopathogenic fungi. The objective of this study was to use B. bassiana M130 as an insecticidal agent against the greenhouse whitefly. M130 isolated from infected insects is known to be a biocontrol agent against greenhouse whitefly. Phylogenetic classification of M130 was determined according to its morphological features and 18S rRNA sequence analysis. M130 was identified as B. bassiana M130 and showed chitinase (342.28 units/ml) and protease (461.70 units/ml) activities, which were involved in the invasion of the host through the outer cuticle layer, thus killing them. The insecticidal activity was 55.2% in petri-dish test, 84.6% in pot test, and 45.3% in field test. The results of this study indicate that B. bassiana has potential as a biological agent for the control of greenhouse whitefly to replace chemical pesticides.

  4. Establishment of fungal entomopathogens Beauveria bassiana and Bionectria ochroleuca (Ascomycota: Hypocreales) as endophytes on artichoke Cynara scolymus.

    Science.gov (United States)

    Guesmi-Jouini, J; Garrido-Jurado, I; López-Díaz, C; Ben Halima-Kamel, M; Quesada-Moraga, E

    2014-06-01

    Entomopathogenic fungi (EPF) are commonly found in diverse habitats and are known to cause mycoses in many different taxa of arthropods. Various unexpected roles have been recently reported for fungal entomopathogens, including their presence as fungal endophytes, plant disease antagonists, rhizosphere colonizers and plant growth promoting fungi. In Tunisia, a wide range of indigenous EPF isolates from different species, such as Beauveria bassiana and Bionectria ochroleuca, were found to occur in the soil, and to be pathogenic against the artichoke aphid Capitophorus elaeagni (Hemiptera: Aphididae). Since endophytic fungi are recently regarded as plant-defending mutualists and their presence in internal plant tissue has been discussed as an adaptive protection against insects, we were interested on elucidating the possible endophytic behavior of B. bassiana and B. ochroleuca on artichoke, Cynara scolymus, after foliar spraying tehcnique. The leaf spray inoculation method was effective in introducing the inoculated fungi into the plant tissues and showed, then, an endophytic activity on artichoke even 10 days later. According S-N-K test, there was significant differences between the two fungal treatments, B. ochroleuca (84% a) and B. bassiana (78% a), and controls (0% b). Likewise, the inoculated entomopathogenic fungi were also isolated from new leaves even though with significant differences respectively between controls (0% c), B. bassiana (56% b) and B. ochroleuca (78% a). These results reveals significant new data on the interaction of inoculated fungi with artichoke plant as ecological roles that can be exploited for the protection of plants.

  5. [Phospholipase, proteinase and hemolytic activities of Candida albicans isolates obtained from clinical specimens].

    Science.gov (United States)

    Yenişehirli, Gülgün; Bulut, Yunus; Tunçoglu, Ebru

    2010-01-01

    This study was aimed to determine the phospholipase, proteinase and hemolytic activities of Candida albicans strains isolated from clinical specimens. A total of 147 C. albicans strains isolated from blood (n = 29), respiratory specimens (n = 44), urine (n = 52), pus (n = 17) and stool (n = 5) were included in the study. Proteinase and phospholipase activities were determined in 81% and 76% of C. albicans isolates, respectively. All C. albicans isolates revealed beta-hemolytic activity on Sabouraud dextrose agar supplemented with 7% fresh sheep blood and 3% glucose. Phospholipase and proteinase positivity were highest among the respiratory isolates. Proteinase activity of respiratory (93%) and blood (83%) isolates were statistically significantly higher than that of urine (77%; p = 0.032), pus (65%; p = 0.007) and stool isolates (60%; p = 0.026). While phospholipase activity showed statistically significant difference between respiratory (84%) and pus (53%) isolates (p = 0.014), no statistically significant difference was determined for blood (79%), urine (75%) and stool (80%) isolates (p > 0.05). Two blood isolates with 4+ proteinase activity and 3 urine isolates with 3+ proteinase activity were phospholipase negative. One urine isolate with 4+ phospholipase activity and 4 with 3+ phospholipase activity were proteinase negative. Phospholipase and proteinase negative 1 isolate from stool and 1 isolate from pus were found to have 4+ hemolytic activity. In conclusion, besides proteinase and phospholipase enzyme activities, hemolytic activity may play an important role for the C.albicans infections. The pathogenetic role of these virulence factors should be evaluated by further clinical studies.

  6. Denaturation and in Vitro Gastric Digestion of Heat-Treated Quinoa Protein Isolates Obtained at Various Extraction pH

    NARCIS (Netherlands)

    Ruiz, Geraldine Avila; Opazo-Navarrete, Mauricio; Meurs, Marlon; Minor, Marcel; Sala, Guido; Boekel, van Martinus; Stieger, Markus; Janssen, Anja E.M.

    2016-01-01

    The aim of this study was to determine the influence of heat processing on denaturation and digestibility properties of protein isolates obtained from sweet quinoa (Chenopodium quinoa Willd) at various extraction pH values (8, 9, 10 and 11). Pretreatment of suspensions of protein isolates at 60,

  7. Denaturation and in Vitro Gastric Digestion of Heat-Treated Quinoa Protein Isolates Obtained at Various Extraction pH

    NARCIS (Netherlands)

    Ruiz, Geraldine Avila; Opazo-Navarrete, Mauricio; Meurs, Marlon; Minor, Marcel; Sala, Guido; Boekel, van Tiny; Stieger, Markus; Janssen, Anja E.M.

    2016-01-01

    The aim of this study was to determine the influence of heat processing on denaturation and digestibility properties of protein isolates obtained from sweet quinoa (Chenopodium quinoa Willd) at various extraction pH values (8, 9, 10 and 11). Pretreatment of suspensions of protein isolates at 60,

  8. Molecular analysis of Mycobacterium isolates from extrapulmonary specimens obtained from patients in Mexico

    Directory of Open Access Journals (Sweden)

    Portillo-Gómez Leopoldo

    2009-03-01

    Full Text Available Abstract Background Little information is available on the molecular epidemiology in Mexico of Mycobacterium species infecting extrapulmonary sites in humans. This study used molecular methods to determine the Mycobacterium species present in tissues and body fluids in specimens obtained from patients in Mexico with extrapulmonary disease. Methods Bacterial or tissue specimens from patients with clinical or histological diagnosis of extrapulmonary tuberculosis were studied. DNA extracts from 30 bacterial cultures grown in Löwenstein Jensen medium and 42 paraffin-embedded tissues were prepared. Bacteria were cultured from urine, cerebrospinal fluid, pericardial fluid, gastric aspirate, or synovial fluid samples. Tissues samples were from lymph nodes, skin, brain, vagina, and peritoneum. The DNA extracts were analyzed by PCR and by line probe assay (INNO-LiPA MYCOBACTERIA v2. Innogenetics NV, Gent, Belgium in order to identify the Mycobacterium species present. DNA samples positive for M. tuberculosis complex were further analyzed by PCR and line probe assay (INNO-LiPA Rif.TB, Innogenetics NV, Gent, Belgium to detect mutations in the rpoB gene associated with rifampicin resistance. Results Of the 72 DNA extracts, 26 (36.1% and 23 (31.9% tested positive for Mycobacterium species by PCR or line probe assay, respectively. In tissues, M. tuberculosis complex and M. genus were found in lymph nodes, and M. genus was found in brain and vagina specimens. In body fluids, M. tuberculosis complex was found in synovial fluid. M. gordonae, M. smegmatis, M. kansasii, M. genus, M. fortuitum/M. peregrinum complex and M. tuberculosis complex were found in urine. M. chelonae/M. abscessus was found in pericardial fluid and M. kansasii was found in gastric aspirate. Two of M. tuberculosis complex isolates were also PCR and LiPA positive for the rpoB gene. These two isolates were from lymph nodes and were sensitive to rifampicin. Conclusion 1 We describe the

  9. Genetic diversity of Japanese encephalitis virus isolates obtained from the Indonesian archipelago between 1974 and 1987.

    Science.gov (United States)

    Schuh, Amy J; Guzman, Hilda; Tesh, Robert B; Barrett, Alan D T

    2013-07-01

    Five genotypes (GI-V) of Japanese encephalitis virus (JEV) have been identified, all of which have distinct geographical distributions and epidemiologies. It is thought that JEV originated in the Indonesia-Malaysia region from an ancestral virus. From that ancestral virus GV diverged, followed by GIV, GIII, GII, and GI. Genotype IV appears to be confined to the Indonesia-Malaysia region, as GIV has been isolated in Indonesia from mosquitoes only, while GV has been isolated on three occasions only from a human in Malaysia and mosquitoes in China and South Korea. In contrast, GI-III viruses have been isolated throughout Asia and Australasia from a variety of hosts. Prior to this study only 13 JEV isolates collected from the Indonesian archipelago had been studied genetically. Therefore the sequences of the envelope (E) gene of 24 additional Indonesian JEV isolates, collected throughout the archipelago between 1974 and 1987, were determined and a series of molecular adaptation analyses were performed. Phylogenetic analysis indicated that over a 14-year time span three genotypes of JEV circulated throughout Indonesia, and a statistically significant association between the year of virus collection and genotype was revealed: isolates collected between 1974 and 1980 belonged to GII, isolates collected between 1980 and 1981 belonged to GIV, and isolates collected in 1987 belonged to GIII. Interestingly, three of the GII Indonesian isolates grouped with an isolate that was collected during the JE outbreak that occurred in Australia in 1995, two of the GIII Indonesian isolates were closely related to a Japanese isolate collected 40 years previously, and two Javanese GIV isolates possessed six amino acid substitutions within the E protein when compared to a previously sequenced GIV isolate collected in Flores. Several amino acids within the E protein of the Indonesian isolates were found to be under directional evolution and/or co-evolution. Conceivably, the tropical climate

  10. Proteases (caseinase and elastase, hemolysins, adhesion and susceptibility to antimicrobials of Stenotrophomonas maltophilia isolates obtained from clinical specimens

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    Garcia Doroti de Oliveira

    2002-01-01

    Full Text Available Forty-six S. maltophilia isolates obtained from hospital clinical specimens were studied for protease (caseinase and elastase production, hemolytic activity, adhesion to HEp-2 cells, plastic and glass. Susceptibility to antimicrobial agents was also evaluated. The majority of isolates were obtained from respiratory tract secretions of patients using medical devices. All the isolates grown overnight were able to hydrolyze casein at 30masculineC and 37masculineC. After 72h, all the isolates hydrolyzed elastase at 30masculineC and 40 isolates (87% at 37masculineC. Most of the isolates presented hemolytic activity after 96h of incubation at both temperatures. Rabbit blood showed the hightest hemolytic activity, after 96h 61% and 98% of tested isolates presented beta-hemolysis at 30masculineC and 37masculineC, respectively. All isolates were susceptible to trimethoprim-sulfametoxazole and were resistant to most beta-lactams tested. By the dilution method, S. maltophilia showed a high susceptibility to ticarcillin-clavulanate and a lower susceptibility to ciprofloxacin than the agar diffusion. The isolates showed adhesion to HEp-2 cells, plastic and glass. The proteolytic activities and adhesion to inanimate surfaces detected in S. maltophilia can be related to the pathogenesis of this bacterium and/or medical device colonization which favors the development of nosocomial infections.

  11. Molecular characterization of Pasteurella multocida isolates obtained from poultry, ruminant, cats and dogs using RAPD and REP-PCR analysis

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    Hesamaddin Shirzad-Aski

    2016-09-01

    Full Text Available In the present study, Randomly Amplified Polymorphic DNA‌ (RAPD and Repetitive Extragenic Palindromic sequence-based Polymerase Chain Reaction (REP-PCR were used to characterize 131 isolates of Pasteurella multocida, originating from different healthy and diseased animal species obtained from several geographical regions of Iran. The RAPD and REP-PCR generated amplified products in the range of 300 to 3400 bp and 200 to 2850 bp, respectively. Among all of the P. multocida isolates, cluster analysis revealed that 63 clusters and nine untypable isolates and 81 clusters and six untypable isolates were produced with RAPD and REP-PCR methods, respectively. The results indicated that the REP-PCR method showed a slightly higher level of discrimination power in differentiating of P. multocida isolates as compared with RAPD. The results showed that a considerable level of genetic diversity exists among P. multocida isolates even in the isolates with the same animal or geographical origins. There was no host- and region-specific pattern. In addition, the isolates obtained from the healthy and diseased animal did not reveal any correlation genotypic profiles, which could be supported by the hypothesis that P. multocida is a strictly opportunistic pathogen. In conclusion, because of a large amount of genetic heterogeneityin the P. multocida isolates, Pasteurellosis may be caused by different clones in the same herd or animal.

  12. Assignment of serotype to Salmonella enterica isolates obtained from poultry and their environment in Southern Brazil.

    Science.gov (United States)

    To assess diversity of Salmonella enterica serotypes present in poultry and their environment from Southern Brazil, the Kauffman-White-LeMinor (KWL) scheme was used to serotype a total of 155 isolates. Isolates were then re-examined with nested PCR and sequencing of the dkgB-linked Intergenic Sequ...

  13. Identification, Characterization and Antibiotic Resistance of Bacterial Isolates Obtained from Waterpipe Device Hoses

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    Majed M. Masadeh

    2015-05-01

    Full Text Available The general lack of knowledge about the health effects of waterpipe smoking is among the reasons for its global spread. In this study, bacterial contamination of waterpipe hoses was investigated. Twenty hoses were collected from waterpipe cafés and screened for bacterial pathogens using standard culture and isolation techniques. Additionally, resistance of isolated bacteria to common antibiotics was determined by identifying the minimum inhibitory concentration (MIC of each isolate. Forty eight bacterial isolates were detected. Isolates included both Gram-positive and Gram-negative pathogens from species that included Micrococcus (12, Corynebacterium (13 and Bacillus (9. In addition, some of the detected pathogens were found to be resistant to aztreonam (79%, cefixime (79%, norfloxacin, amoxicillin (47%, clarithromycin (46% and enrofloxacin (38%. In conclusion, the hose of the waterpipe device is a good environment for the growth of bacterial pathogens, which can then be transmitted to users.

  14. Identification, characterization and antibiotic resistance of bacterial isolates obtained from waterpipe device hoses.

    Science.gov (United States)

    Masadeh, Majed M; Hussein, Emad I; Alzoubi, Karem H; Khabour, Omar; Shakhatreh, Muhamad Ali K; Gharaibeh, Mahmoud

    2015-05-13

    The general lack of knowledge about the health effects of waterpipe smoking is among the reasons for its global spread. In this study, bacterial contamination of waterpipe hoses was investigated. Twenty hoses were collected from waterpipe cafés and screened for bacterial pathogens using standard culture and isolation techniques. Additionally, resistance of isolated bacteria to common antibiotics was determined by identifying the minimum inhibitory concentration (MIC) of each isolate. Forty eight bacterial isolates were detected. Isolates included both Gram-positive and Gram-negative pathogens from species that included Micrococcus (12), Corynebacterium (13) and Bacillus (9). In addition, some of the detected pathogens were found to be resistant to aztreonam (79%), cefixime (79%), norfloxacin, amoxicillin (47%), clarithromycin (46%) and enrofloxacin (38%). In conclusion, the hose of the waterpipe device is a good environment for the growth of bacterial pathogens, which can then be transmitted to users.

  15. Bioesai bioinsektisida Beauveria bassiana dari Sumatera Selatan terhadap kutu putih pepaya, Paracoccus marginatus Williams & Granara De Willink (Hemiptera: Pseudococcidae

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    Siti Herlinda

    2015-09-01

    Full Text Available In fresh swamp areas of South Sumatra, papaya mealybug, Paracoccus marginatus (Hemiptera: Pseudococcidae could cause severe damage to young papaya trees and decreased the fruit quality. The objective of this research was to bioassay test of the bioinsecticide Beauveria bassiana againts nymphs of P. marginatus. B. bassiana was conducted on rice medium. The bioinsecticide was formulated using dried compost, compost gram, paddy ash, paddy bran, woody powder, paddy bran mixed with woody powder, compost enriched with Trichoderma virens. Controls used were sterile water (control 1 and isolate of B. bassiana (control 2. Results showed that conidial viability of B. bassiana on control 2 was the highest (46.33%. The viability of control 2 was not significantly different from the formulations with carrier of the paddy bran, the paddy bran mixed with woody powder, and the compost enriched with T. virens. The highest nymph mortality (82.86% was found on formulation of compost enriched with T. virens and was significantly different from other treatments. The lowest visibly (73.48% occured on formulation of paddy ash, and was significantly different from other treatments. Mortality on control 1 on average was 29.52%, whereas control 2 averaged of 75.71%. The shortest median lethal time (LT50 (3.55 days was found on formulation of compost enriched with T. virens but the longest one (3.73 days occured on the formulation of paddy ash. Overall, the most effective bioinsecticide was the formulation of compost enriched with T. virens.

  16. Antimicrobial resistance trends among Salmonella isolates obtained from horses in the northeastern United States (2001-2013).

    Science.gov (United States)

    Cummings, Kevin J; Perkins, Gillian A; Khatibzadeh, Sarah M; Warnick, Lorin D; Aprea, Victor A; Altier, Craig

    2016-05-01

    OBJECTIVE To describe the antimicrobial resistance patterns of Salmonella isolates obtained from horses in the northeastern United States and to identify trends in resistance to select antimicrobials over time. SAMPLE 462 Salmonella isolates from horses. PROCEDURES Retrospective data were collected for all Salmonella isolates obtained from equine specimens that were submitted to the Cornell University Animal Health Diagnostic Center between January 1, 2001, and December 31, 2013. Temporal trends in the prevalence of resistant Salmonella isolates were investigated for each of 13 antimicrobials by use of the Cochran-Armitage trend test. RESULTS The prevalence of resistant isolates varied among antimicrobials and ranged from 0% (imipenem) to 51.5% (chloramphenicol). During the observation period, the prevalence of resistant isolates decreased significantly for amoxicillin-clavulanic acid, ampicillin, cefazolin, cefoxitin, ceftiofur, chloramphenicol, and tetracycline and remained negligible for amikacin and enrofloxacin. Of the 337 isolates for which the susceptibility to all 13 antimicrobials was determined, 138 (40.9%) were pansusceptible and 192 (57.0%) were multidrug resistant (resistant to ≥ 3 antimicrobial classes). The most common serovar isolated was Salmonella Newport, and although the annual prevalence of that serovar decreased significantly over time, that decrease had only a minimal effect on the observed antimicrobial resistance trends. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that current antimicrobial use in horses is not promoting the emergence and dissemination of antimicrobial-resistant Salmonella strains in the region served by the laboratory.

  17. Identification of xanthans isolated from sugarcane juices obtained from scalded plants infected by Xanthomonas albilineans.

    Science.gov (United States)

    Fontaniella, Blanca; Rodríguez, C W; Piñón, Dolores; Vicente, C; Legaz, María-Estrella

    2002-04-25

    The exudate gum produced by Xanthomonas albilineans, a specific sugarcane pathogen, has been isolated from juices of diseased sugarcane stalks, hydrolyzed with hydrochloric acid, and the hydrolysate analyzed by capillary electrophoresis. Sucrose. cellobiose, mannose, glucose, glucose-1-P and glucuronic acid were identified as the major components of the polysaccharide isolated from diseased stalks. Juices from healthy stalks contained maltose instead of cellobiose. The chemical nature of this polysaccharide is discussed.

  18. Sensitivity of Beauveria bassiana conidia to abiotic factors after successive in vitro subcultures

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    Janaina Zorzetti

    2014-09-01

    Full Text Available The aim of this study was to evaluate the effect of in vitro successive subcultures of Beauveria bassiana isolates in different nutritional conditions on conidia heat and UV radiation tolerance and on sensitivity to azoxystrobin fungicide. The fungus was initially inoculated in Alphitobius diaperinus (Coleoptera: Tenebrionidae adults and after its development, the conidia were cultivated in PDA (potato dextrose agar and MPE (medium to produce Beauveria spp. conidia originating the first subculture 1st (A conidia, which were successively multiplied in their media until the 20th cultivation. Conidia taken from the 20th subculture were again inoculated in the insect and after sporulation they were once more multiplied in the media, producing the 1st (B conidia. For the tests, 1st (A, 10th, 20th and 1st (B conidia from the CG 152 and Unioeste 40 isolates were used. Successive subcultures and the medium nutritional conditions interfered on the fungus physiology, and the isolates showed different behaviors. The subcultures on PDA showed higher thermotolerance to Unioeste 40 in all subcultures and to CG 152 just after inoculation in the host. B. bassiana was susceptible to UV radiation, and this characteristic was influenced by the subcultures in different media. Both isolates were more tolerant to UV radiation when subcultivated on MPE, while just for the 40 Unioeste this tolerance increased after host inoculation. The B. bassiana sensitivity to azoxystrobin fungicide was affected after successive subcultures in both medium. Conidia originated in MPE were more resistant to fungicide and the host inoculation increased this tolerance. These results show that it is possible to enhance the fungi efficiency to control pests, through the selection of a virulent isolate, and handling their nutritional conditions during cultivation and host inoculation.

  19. 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

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    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

  20. Genome Sequences of 12 Bacterial Isolates Obtained from the Urine of Pregnant Women

    Science.gov (United States)

    Weimer, Cory M.; Deitzler, Grace E.; Robinson, Lloyd S.; Park, SoEun; Hallsworth-Pepin, Kymberlie; Wollam, Aye; Mitreva, Makedonka

    2016-01-01

    The presence of bacteria in urine can pose significant risks during pregnancy. However, there are few reference genome strains for many common urinary bacteria. We isolated 12 urinary strains of Streptococcus, Staphylococcus, Citrobacter, Gardnerella, and Lactobacillus. These strains and their genomes are now available to the research community. PMID:27688327

  1. Biofilm Formation by Drug Resistant Enterococci Isolates Obtained from Chronic Periodontitis Patients

    Science.gov (United States)

    Mehta, Manjula; Sood, Shaveta; Sharma, Jyoti

    2017-01-01

    Introduction Enterococci are an important cause of opportunistic nosocomial infections and several multidrug resistant strains have emerged. The severity of periodontal diseases is managed by reduction in the pathogenic bacteria. There is a need to assess the prevalence and antibiotic susceptibility of enterococci colonizing the periodontal pocket and correlate its biofilm formation ability because oral biofilms provide a protective environment and are a reservoir of bacterial colonization of the gingival crevice. Aim To investigate possible association between antibiotic susceptibility and biofilm formation in enterococci isolates from chronic periodontitis patients. Materials and Methods This retrospective study was conducted at Dr. Harvansh Singh Judge Institute of Dental Sciences and Hospital, Punjab University, Chandigarh from January 2015 to October 2015. Sterile paper points were inserted in the periodontal pocket of 100 subjects and put in a transport media. Forty -six isolates were identified as enterococci. The isolates were further examined for their ability to form biofilm by microtitre plate assay and antimicrobial susceptibility testing was done by disc diffusion method for clinically relevant antibiotics. Results Significant relationship (p<0.001) was found between biofilm production with antibiotic resistance to Vancomycin, Erythromycin, Ciprofloxacin, Tiecoplanin, Amoxycillin and Gentamycin. Conclusion The study demonstrates a high propensity among the isolates of Enterococci to form biofilm and a significant association of biofilm with multiple drug resistance. PMID:28273964

  2. Characterization of Enterobacteriaceae isolates obtained from a tertiary care hospital in Mexico, which produces extended-spectrum β-lactamase.

    Science.gov (United States)

    Morfín-Otero, Rayo; Mendoza-Olazarán, Soraya; Silva-Sánchez, Jesús; Rodríguez-Noriega, Eduardo; Laca-Díaz, Jorge; Tinoco-Carrillo, Perla; Petersen, Luis; López, Perla; Reyna-Flores, Fernando; Alcantar-Curiel, Dolores; Garza-Ramos, Ulises; Garza-González, Elvira

    2013-10-01

    The prevalence and genetic characteristics of Escherichia coli and Klebsiella pneumoniae clinical isolates producing extended-spectrum β-lactamase (ESBL) were examined. Between October 2010 and March 2011, E. coli (n=460) and K. pneumoniae (n=78) isolates were collected at a tertiary care hospital in Guadalajara, Mexico. The minimum inhibitory concentration (MIC) for each isolate was determined using a broth microdilution method, and ESBL production was assayed. The presence of β-lactamase genes, blaSHV, blaCTX-M, and blaTLA-1, was detected by PCR and confirmed with sequencing. Only ESBL-producing isolates were further subjected to pulsed-field gel electrophoresis (PFGE) and plasmid profiling. All of the ESBL isolates were multidrug resistant and 75/460 (16.3%) E. coli isolates and 21/78 (26.9%) K. pneumoniae isolates were found to produce ESBL. For the E. coli isolates, >95% susceptibility to amikacin, meropenem, fosfomycin, imipenem, and nitrofurantoin was observed. For K. pneumoniae, similar results were obtained, with discrepancies observed for gentamicin and nitrofurantoin. PFGE further identified eleven pulsotypes for E. coli and three clusters of K. pneumoniae. CTX-M-15 was detected in 85% of ESBL-producing E. coli and in 76% of ESBL-producing K. pneumoniae. In contrast, SHV-5 ESBL was identified in 17% of E. coli isolates and in 86% of K. pneumoniae isolates. The bla-TLA-1 gene was not detected in any of the 96 isolates analyzed. Overall, CTX-M-15 and SHV-5 were found to have a high rate of spread throughout the hospital and were associated with strong multidrug resistance.

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

    Science.gov (United States)

    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.

  4. Laboratory evaluation of a native strain of Beauveria bassiana for controlling Dermanyssus gallinae (De Geer, 1778) (Acari: Dermanyssidae).

    Science.gov (United States)

    Immediato, Davide; Camarda, Antonio; Iatta, Roberta; Puttilli, Maria Rita; Ramos, Rafael Antonio Nascimento; Di Paola, Giancarlo; Giangaspero, Annunziata; Otranto, Domenico; Cafarchia, Claudia

    2015-09-15

    The poultry red mite, Dermanyssus gallinae (De Geer, 1778) (Acari: Dermanyssidae) is one of the most economically important ectoparasites of laying hens worldwide. Chemical control of this mite may result in environmental and food contamination, as well as the development of drug resistance. High virulence of Beauveria bassiana sensu lato strains isolated from naturally infected hosts or from their environment has been demonstrated toward many arthropod species, including ticks. However, a limited number of studies have assessed the use of B. bassiana for the control of D. gallinae s.l. and none of them have employed native strains. This study reports the pathogenicity of a native strain of B. bassiana (CD1123) against nymphs and adults of D. gallinae. Batches of nymph and adult mites (i.e., n=720 for each stage) for treated groups (TGs) were placed on paper soaked with a 0.1% tween 80 suspension of B. bassiana (CIS, 10(5), 10(7) and 10(9) conidia/ml), whilst 240 untreated control mites for each stage (CG) were exposed only to 0.1% tween 80. The mites in TG showed a higher mortality at all stages (p<0.01) when compared to CG, depending on the time of exposure and the conidial concentration. A 100% mortality rate was recorded using a CIS of 10(9) conidia/ml 12 days post infection (DPI) in adults and 14 DPI in nymphs. B. bassiana suspension containing 10(9) conidia/ml was highly virulent towards nymph and adult stages of D. gallinae, therefore representing a possible promising natural product to be used in alternative or in combination to other acaricidal compounds currently used for controlling the red mite.

  5. Glucose concentration alters dissolved oxygen levels in liquid cultures of Beauveria bassiana and affects formation and bioefficacy of blastospores.

    Science.gov (United States)

    Mascarin, Gabriel Moura; Jackson, Mark A; Kobori, Nilce Naomi; Behle, Robert W; Dunlap, Christopher A; Delalibera Júnior, Ítalo

    2015-08-01

    The filamentous fungus Beauveria bassiana is an economically important pathogen of numerous arthropod pests and is able to grow in submerged culture as filaments (mycelia) or as budding yeast-like blastospores. In this study, we evaluated the effect of dissolved oxygen and high glucose concentrations on blastospore production by submerged cultures of two isolates of B. bassiana, ESALQ1432 and GHA. Results showed that maintaining adequate dissolved oxygen levels coupled with high glucose concentrations enhanced blastospore yields by both isolates. High glucose concentrations increased the osmotic pressure of the media and coincided with higher dissolved oxygen levels and increased production of significantly smaller blastospores compared with blastospores produced in media with lower concentrations of glucose. The desiccation tolerance of blastospores dried to less than 2.6 % moisture was not affected by the glucose concentration of the medium but was isolate dependent. Blastospores of isolate ESALQ1432 produced in media containing 140 g glucose L(-1) showed greater virulence toward whitefly nymphs (Bemisia tabaci) as compared with blastospores produced in media containing 40 g glucose L(-1). These results suggest a synergistic effect between glucose concentration and oxygen availability on changing morphology and enhancing the yield and efficacy of blastospores of B. bassiana, thereby facilitating the development of a cost-effective production method for this blastospore-based bioinsecticide.

  6. Beauveria bassiana keratitis in bullous keratopathy: antifungal sensitivity testing and management.

    Science.gov (United States)

    Figueira, Luís; Pinheiro, Dolores; Moreira, Raul; Pinto, Eugénia; Simões, Joana; Camisa, Elisa; Torrão, Luís; Palmares, Jorge; Falcão-Reis, Fernando

    2012-01-01

    Beauveria bassiana is a ubiquitous fungus available as an insecticide. In humans, it has limited virulence; to our knowledge, only 3 cases of invasive disease and 10 cases of keratitis have been documented. We report the first case of B bassiana keratitis in a patient with aphakic bullous keratopathy. The fungal keratitis proved to be highly resistant to topical clotrimazole. Molecular identification was based on DNA sequence analysis. The minimal inhibitory concentrations (MIC) obtained were 2 µg/mL for voriconazole, 0.250 µg/mL for posaconazole, and >128 µg/mL for fluconazole; amphotericin B MIC was >16 µg/mL. In the absence of clinical improvement, a penetrating keratoplasty (PK) was performed. The patient was discharged on topical and systemic voriconazole and prednisolone 40 mg PO/day. The eye remained calm with a transparent cornea and clear anterior chamber. B bassiana keratitis is extremely rare, with only a few cases reported. Its risk factors are unknown. We report the first case in a patient with aphakic bullous keratopathy, which proved highly resistant to antifungal therapy (antifungal susceptibility results are presented). A PK was necessary for clinical improvement. A review of the literature is performed in an effort to define therapeutic strategies.

  7. Development of a method for detection and quantification of B. brongniartii and B. bassiana in soil

    Science.gov (United States)

    Canfora, L.; Malusà, E.; Tkaczuk, C.; Tartanus, M.; Łabanowska, B. H.; Pinzari, F.

    2016-03-01

    A culture independent method based on qPCR was developed for the detection and quantification of two fungal inoculants in soil. The aim was to adapt a genotyping approach based on SSR (Simple Sequence Repeat) marker to a discriminating tracing of two different species of bioinoculants in soil, after their in-field release. Two entomopathogenic fungi, Beauveria bassiana and B. brongniartii, were traced and quantified in soil samples obtained from field trials. These two fungal species were used as biological agents in Poland to control Melolontha melolontha (European cockchafer), whose larvae live in soil menacing horticultural crops. Specificity of SSR markers was verified using controls consisting of: i) soil samples containing fungal spores of B. bassiana and B. brongniartii in known dilutions; ii) the DNA of the fungal microorganisms; iii) soil samples singly inoculated with each fungus species. An initial evaluation of the protocol was performed with analyses of soil DNA and mycelial DNA. Further, the simultaneous detection and quantification of B. bassiana and B. brongniartii in soil was achieved in field samples after application of the bio-inoculants. The protocol can be considered as a relatively low cost solution for the detection, identification and traceability of fungal bio-inoculants in soil.

  8. Effects of morning and night application of Beauveria bassiana strains NI8 and GHA against the tarnished plant bug in cotton

    Science.gov (United States)

    The tarnished plant bug, (TPB), Lygus lineolaris (Palisot de Beauvois), (Hemiptera: Miridae) an important pest of cotton (Gosssypium hirsutum L.) found in the Mississippi Delta is naturally attacked by the entomopathogenic fungus Beauveria bassiana (Balsamo) Vueillemin. In this study, two isolates o...

  9. Obtaining freshly isolated and cultured mesenchymal stem cells from human adipose tissue.

    Science.gov (United States)

    Boquest, Andrew C; Collas, Philippe

    2012-01-01

    The stromal compartment of adipose tissue harbors mesenchymal stem cells (MSCs) (also called stromal stem cells) that display extensive proliferative capacity and multilineage differentiation potential. Such cells offer a practical avenue of generating patient-matched tissue for use in regenerative medicine. It is relatively easy to isolate these cells from adipose tissue in large enough quantities (tens of millions) to allow for their clinical use in a native, uncultured form. Alternatively, MSCs from adipose tissue can be expanded and differentiated into the desired tissue type in vitro using straightforward cell culture techniques. In this chapter, we outline procedures for isolating large numbers of highly purified MSCs from human adipose tissue in their native, uncultured form and methods for their subsequent expansion and differentiation in vitro.

  10. Isolation and Purification of an Early Pregnancy Factor–Like Molecule from Culture Supernatants Obtained from Lymphocytes of Pregnant Women

    OpenAIRE

    1998-01-01

    Purpose:Our purpose was to determine whether lymphocytes synthesize proteins during pregnancy, to observe whether one of the proteins synthesized has early pregnancy factor (EPF)–like activity and to isolate and purify this molecule from culture supernatants obtained from stimulated lymphocytes of pregnant women.

  11. 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

    Directory of Open Access Journals (Sweden)

    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

  12. Virulence attenuation and phenotypic variation of Paracoccidioides brasiliensis isolates obtained from armadillos and patients

    Directory of Open Access Journals (Sweden)

    SAG Macoris

    2006-05-01

    Full Text Available Paracoccidioides brasiliensis is the etiological agent of paracoccidioidomycosis, the most important systemic mycosis in Latin America. The virulence profiles of five isolates of P. brasiliensis were studied in two different moments and correlated with some colonial phenotypic aspects. We observed a significant decrease in the virulence and an intense phenotypic variation in the mycelial colony. The recognition of all ranges of phenotypic and virulence variation of P. brasiliensis, as well as its physiological and genetic basis, will be important for a better comprehension of its pathogenic and epidemiological features.

  13. In vitro and in vivo antitumor activity of crude extracts obtained from Brazilian Chromobacterium sp isolates

    Energy Technology Data Exchange (ETDEWEB)

    Menezes, C.B.A.; Silva, B.P. [Universidade Estadual de Campinas, Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas, Campinas, SP (Brazil); Universidade de São Paulo, Interunidades em Biotecnologia, São Paulo, SP (Brazil); Sousa, I.M.O.; Ruiz, A.L.T.G.; Spindola, H.M. [Universidade Estadual de Campinas, Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas, Campinas, SP (Brazil); Cabral, E.; Eberlin, M.N. [Instituto de Química, Universidade Estadual de Campinas, Laboratório Thomson Mass Spectrometry, Campinas, SP (Brazil); Tinti, S.V.; Carvalho, J.E. [Universidade Estadual de Campinas, Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas, Campinas, SP (Brazil); Foglio, M.A.; Fantinatti-Garboggini, F. [Universidade Estadual de Campinas, Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas, Campinas, SP (Brazil); Universidade de São Paulo, Interunidades em Biotecnologia, São Paulo, SP (Brazil)

    2012-10-23

    Natural products produced by microorganisms have been an important source of new substances and lead compounds for the pharmaceutical industry. Chromobacterium violaceum is a Gram-negative β-proteobacterium, abundant in water and soil in tropical and subtropical regions and it produces violacein, a pigment that has shown great pharmaceutical potential. Crude extracts of five Brazilian isolates of Chromobacterium sp (0.25, 2.5, 25, and 250 µg/mL) were evaluated in an in vitro antitumor activity assay with nine human tumor cells. Secondary metabolic profiles were analyzed by liquid chromatography and electrospray ionization mass spectrometry resulting in the identification of violacein in all extracts, whereas FK228 was detected only in EtCE 308 and EtCE 592 extracts. AcCE and EtCE 310 extracts showed selectivity for NCI/ADR-RES cells in the in vitro assay and were evaluated in vivo in the solid Ehrlich tumor model, resulting in 50.3 and 54.6% growth inhibition, respectively. The crude extracts of Chromobacterium sp isolates showed potential and selective antitumor activities for certain human tumor cells, making them a potential source of lead compounds. Furthermore, the results suggest that other compounds, in addition to violacein, deoxyviolacein and FK228, may be involved in the antitumor effect observed.

  14. 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.

  15. Efficacy of Beauveria bassiana substrates and formulations for the ...

    African Journals Online (AJOL)

    Mo

    Efficacy of Beauveria bassiana substrates and formulations for the control of banana weevil. E. Magara ..... huge challenge to its utilzation in biological control programmes. .... Journal of invertebrate Pathology 38:191-200. Nankinga Calorine.

  16. Equine preantral follicles obtained via the Biopsy Pick-Up method: histological evaluation and validation of a mechanical isolation technique.

    Science.gov (United States)

    Haag, K T; Magalhães-Padilha, D M; Fonseca, G R; Wischral, A; Gastal, M O; King, S S; Jones, K L; Figueiredo, J R; Gastal, E L

    2013-03-15

    The aims of this study in mares were to: (1) compare preantral follicle parameters between in vitro Biopsy Pick-Up (BPU) and scalpel blade collection methods and between histological and mechanical isolation processing (experiment 1); (2) histologically evaluate preantral follicles (experiment 2); and (3) compare histological analysis with a previously established mechanical isolation technique using a tissue chopper (experiment 3) for ovarian cortical fragments obtained in vivo using a BPU instrument. In experiment 1, preantral follicles were analyzed (N = 220; 90% primordial and 10% primary). Proportions of primordial and primary follicles did not differ (P > 0.05) between tissue collection (BPU vs. scalpel blade dissection) or processing (mechanical isolation vs. histology) methods. Follicle viability and morphology rates were similar (P > 0.05) between tissue collection methods, but mechanical isolation produced more (P 0.05) by processing methods. In conclusion, most parameters evaluated for preantral follicles were similar between histological and tissue chopper processing techniques; hence, mechanical isolation efficiently dissociated equine preantral follicles from the ovarian cortex. Therefore, the tissue chopper could be used to isolate large numbers of morphologically normal equine preantral follicles for cryopreservation and/or in vitro culture.

  17. In vitro inflammatory responses elicited by isolates of Alloiococcus otitidis obtained from children with otitis media with effusion.

    Science.gov (United States)

    Ashhurst-Smith, Christopher; Hall, Sharron T; Burns, Christine J; Stuart, John; Blackwell, C Caroline

    2014-04-01

    Alloiococcus otitidis is usually detected in children with otitis media (OM) by PCR as it is not often detected by routine culture. Our improved method for its isolation obtained A. otitidis from nearly 50% of 78 children with OM with effusion. The role of A. otitidis in pathogenesis of OM is unclear. This study tested two hypothesis: (1) that fresh isolates of A. otitidis would elicit pro-inflammatory cytokines from THP-1 monocytic cells equivalent to those induced by Streptococcus pneumoniae; (2) priming THP-1 cells with interferon-gamma (IFN-γ) a surrogate for virus infection, would enhance pro-inflammatory responses. Recent clinical isolates of A. otitidis, S. pneumoniae (ATCC 49619) and a blood culture isolate of S. pneumoniae (SP2) were used in the assays. Cytokines were quantified by BioRad bead assay and Luminex 200. IFN-γ priming enhanced cytokine responses. S. pneumoniae ATCC 49619 induced lower responses than SP2 for IL-1β, IL-6, TNF-α. A. otitidis LW 27 elicited higher IL-1β and TNF-α responses than either pneumococcal isolate. Small green colony types of A. otitidis induced higher responses than large white colony types for IL-8 and IL-1β. The hypothesis that A. otitidis elicits cytokines observed in middle ear effusions was supported; the need to use recent clinical isolates in studies of pathogenesis was highlighted.

  18. [A Large Number of Circulating Tumor Cells(CTCs)Can Be Isolated from Samples Obtained by Using Leukapheresis Procedures].

    Science.gov (United States)

    Soya, Ryoko; Taguchi, Jyunichi; Nagakawa, Yuichi; Takahashi, Osamu; Sandoh, Norimasa; Hosokawa, Yuichi; Kasuya, Kazuhiko; Umeda, Naoki; Okamoto, Masato; Tsujitani, Shunichi; Tsuchida, Akihiko

    2015-09-01

    We hypothesized that a large number of circulating tumor cells(CTCs)may be isolated from samples obtained by using the leukapheresis procedures that are utilized to collect peripheral blood mononuclear cells for dendritic cell vaccine therapy. We utilized the CellSearch System to determine the number of CTCs in samples obtained by using leukapheresis in 7 patients with colorectal cancer, 5 patients with breast cancer, and 3 patients with gastric cancer. In all patients, a large number of CTCs were isolated. The mean number of CTCs per tumor was 17.1(range 10-34)in colorectal cancer, 10.0(range 2-27)in breast cancer, and 24.0(range 2-42)in gastric cancer. We succeeded in culturing the isolated CTCs from 7 patients with colorectal cancer, 5 patients with breast cancer, and 3 patients with gastric cancer. In conclusion, compared to conventional methods, a large number of CTCs can be obtained by using leukapheresis procedures. The molecular analyses of the CTCs isolated by using this method should be promising in the development of personalized cancer treatments.

  19. Successful treatment of Beauveria bassiana fungal keratitis with topical voriconazole.

    Science.gov (United States)

    Ogawa, Akiko; Matsumoto, Yukihiro; Yaguchi, Takashi; Shimmura, Shigeto; Tsubota, Kazuo

    2016-04-01

    We describe a 66-year-old woman who suffered from fungal keratitis after corneal transplantation. The causative organism was identified as Beauveria bassiana on the basis of morphological characteristics and the sequence of the internal transcribed spacer region of the ribosomal RNA gene. The patient was successfully treated with topical voriconazole (VRCZ) use only. We, hereby, present the first report of a case with B. bassiana fungal keratitis that responded to topical antifungal VRCZ treatment.

  20. Identification and Characterization of Imipenem-Resistant Klebsiella pneumoniae and Susceptible Klebsiella variicola Isolates Obtained from the Same Patient.

    Science.gov (United States)

    Garza-Ramos, Ulises; Moreno-Dominguez, Stephania; Hernández-Castro, Rigoberto; Silva-Sanchez, Jesús; Barrios, Humberto; Reyna-Flores, Fernando; Sanchez-Perez, Alejandro; Carrillo-Casas, Erika M; Sanchez-León, María Carmen; Moncada-Barron, David

    2016-04-01

    Klebsiella variicola, a bacterium closely genetically related to Klebsiella pneumoniae, is commonly misidentified as K. pneumoniae by biochemical tests. To distinguish between the two bacteria, phylogenetic analysis of the rpoB gene and the identification of unique genes in both bacterial species by multiplex-polymerase chain reaction (PCR) provide the means to reliably identify and genotype K. variicola. In recent years, K. variicola has been described both as the cause of an intrahospital outbreak in a pediatric hospital, which resulted in sepsis in inpatients, and as a frequent cause of bloodstream infections. In the present study, K. pneumoniae and K. variicola were isolated from a unique patient displaying different antimicrobial susceptibility phenotypes and different genotypes of virulence determinants. Eight clinical isolates were obtained at different time intervals; all during a 5-month period. The isolates were identified as K. pneumoniae by an automated identification system. The clinical (biochemical test) and molecular (multiplex-PCR and rpoB gene) characterization identified imipenem resistance in the first six K. pneumoniae ST258 isolates, which encode the SHV-12 cephalosporinase and KPC-3 carbapenemase genes. The two last remaining isolates corresponded to susceptible K. variicola. The bacterial species showed a specific profile of virulence-associated determinants, specifically the fimA, fimH, and ecpRAB fimbrial-encoding genes identified only in K. pneumoniae isolates. However, the entb (enterobactin), mrkD (fimbrial adhesin), uge (epimerase), ureA (urease), and wabG (transferase) genes were shared between both bacterial species. Recent studies attribute a higher mortality rate to K. variicola than to K. pneumonia. This work highlights the identification of K. pneumoniae and the closely related K. variicola isolated from the same patient. The value of distinguishing between these two bacterial species is in their clinical significance, their

  1. CAMP-reaction among skin isolates obtained from a dog with an acute squamous eczema.

    Science.gov (United States)

    Brückler, J; Wibawan, I W; Lämmler, C

    1990-12-01

    The primary culture of a clinical specimen obtained from a dog with an acute squamous eczema revealed 3 different bacterial cultures. Two of these cultures, a beta-hemolytic Staphylococcus aureus and a group B streptococcal culture, demonstrated synergistic hemolytic activities on this primary culture plate. The group B streptococcus had the serotype surface antigens Ib/c, protein antigen c in its c beta component.

  2. Isolation and characterization of starch obtained from Brosimum alicastrum Swarts seeds.

    Science.gov (United States)

    Pérez-Pacheco, E; Moo-Huchin, V M; Estrada-León, R J; Ortiz-Fernández, A; May-Hernández, L H; Ríos-Soberanis, C R; Betancur-Ancona, D

    2014-01-30

    In this paper, the Ramon starch was isolated and its chemical composition and physical and microscopic characteristics were determined. Corn starch was used as reference. In general, the proximal composition was similar between starches studied. Ramon starch granules were oval-spherical and rounded with sizes between 6.5 and 15 μm. Starch purity was high (92.57%) with amylose content of 25.36%. The gelatinization temperature was 83.05°C and transition enthalpy was 21.423 J/g. At 90°C, solubility was 20.42%, swelling power 17.64 g water/gstarch and water absorption capacity was 13 gwater/gstarch. The pH, clarity and color (Hue angle) of Ramon starch were higher to those reported for corn starch. The results achieved suggest that Ramon starch has potential for application in food systems requiring high processing temperatures and it is also a promising option for use in the manufacture of biodegradable materials.

  3. Isolation, expansion and differentiation of cellular progenitors obtained from dental pulp of agouti (Dasyprocta prymnolopha Wagler, 1831

    Directory of Open Access Journals (Sweden)

    Yulla K.P. de Carvalho

    2015-06-01

    Full Text Available Abstract: The study aimed to isolate, expand, differentiate and characterize progenitor cells existent in the dental pulp of agouti. The material was washed with PBS solution and dissociated mechanically with the aid of a scalpel blade on plates containing culture medium D-MEM/F-12, and incubated at 5% CO2-37⁰C. The growth curve, CFU assay, osteogenic/adipogenic differentiation and characterization were obtained from the isolation. The cells began to be released from the explant tissue around the 7th day of culture. By day 22 of culture, cells reached 80% confluence. At the UFC test, 81 colonies were counted with 12 days of cultivation. The growth curves before and after freezing showed a regular growth with intense proliferation and clonogenic potential. The cell differentiation showed formation of osteoblasts and fat in culture, starting at 15 days of culture in a specific medium. Flow cytometry (FACs was as follows: CD34 (positive, CD14 (negative, CD45 (negative, CD73 (positive, CD79 (negative, CD90 (positive, CD105 (positive, demonstrating high specificity and commitment of isolated cells with mesenchymal stem cells strains. These results suggest the existence of a cell population of stem cells with mesenchymal features from the isolated tissue in the explants of agouti dental pulp, a potential model for study of stem cell strains obtained from the pulp tissue.

  4. Laboratory Study on Biological Control of Ticks (Acari: Ixodidae) by Entomopathogenic Indigenous Fungi (Beauveria bassiana)

    OpenAIRE

    M Abdigoudarzi; Esmaeilnia, K; Shariat, N

    2009-01-01

    Background: Chemical control method using different acaricides as spray, dipping solution or pour-on is routinely used for controlling ticks. Biological control agents are favorable due to their safety for animals and environment. Entomopathogenic fungi such as Beauveria bassiana are well known for controlling ticks. In this study, two Iranian indigenous strains of B. bassiana (B. bassiana 5197 and B. bassiana Evin) were selected and grown on specific me­dia. The pathogenic effects of the...

  5. PCR detection and serotyping of enterotoxigenic and shigatoxigenic Escherichia coli isolates obtained from chicken meat in Mumbai, India

    Directory of Open Access Journals (Sweden)

    R. J. Zende,

    2013-08-01

    Full Text Available Aim: Present study was undertaken to find out the frequency of few virulent genes and prevalence of related strains of Escherichia coli isolated from chicken meat obtained from chicken retail shops by Polymerase Chain Reaction (PCR.Materials and Methods: 66 samples of freshly slaughtered chicken meat were collected from 22 identified retail shops located at Mumbai city, randomly. Processed meat samples were cultured in EMB agar and presumptive colonies were confirmed by various biochemical tests. PCR method was accustomed for identification of the genes coding for heat-stable enterotoxin a (STa, heat labile enterotoxin (LT, shiga-like toxins 1 and 2 (SLT1 and SLT2. E. coli isolates were sent to National Salmonella and Escherichia Centre, CRI, Kasauli, HP, India for serotyping.Results: 11 (16.67% E. coli strains were isolated from 66 chicken meat samples. 3 (27.27% out of 11 harbored the gene for SLT2, and 2 (18.18% for STa. None of the strain contains SLT1 and LT genes. Serotypes detected were rough, O2, O20, O22, O102 each for one isolate and 6 isolates were untypable (UT.Conclusion: The results concluded that chicken meat samples analysed harbored genes for shiga like toxins and enterotoxins and different serotypes of E. coli. These findings indicating that regular monitoring of chicken meat is essential for this pathogen to prevent potential public health problems.

  6. Anti-Trichomonas vaginalis activity of Hypericum polyanthemum extract obtained by supercritical fluid extraction and isolated compounds.

    Science.gov (United States)

    Cargnin, Simone Tasca; Vieira, Patrícia de Brum; Cibulski, Samuel; Cassel, Eduardo; Vargas, Rubem Mário Figueiró; Montanha, Jarbas; Roehe, Paulo; Tasca, Tiana; von Poser, Gilsane Lino

    2013-04-01

    The anti-Trichomonas vaginalis activity of Hypericum polyanthemum extract obtained by supercritical fluid extraction (50°C, 150bar) and the chemical compounds isolated and purified from this extract (benzopyrans HP1, HP2, HP3, and phloroglucinol derivative uliginosin B) were assessed. All samples had anti-T. vaginalis activity; however, HP1 demonstrated the best selectivity against this protozoan (metronidazole-resistant and susceptible isolates), with no cytotoxicity on mammalian cells (selectivity index of 73.97). Moreover, HP1 had activity against a metronidazole-resistant isolate (52% of viable trophozoites), and this effect was higher when tested with a low concentration of metronidazole (23% of viable trophozoites). Experiments demonstrated that all isolated compounds caused damage to the parasites' membrane (>90% of LDH release) and do not present a notable hemolytic effect, although HP2 and uliginosin B exhibited cytotoxicity against mammalian cells. Therefore, the analyzed molecules are promising prototypes for new antiprotozoal drugs, especially HP1, which seems to improve metronidazole's effect on a resistant T. vaginalis isolate.

  7. A method for obtaining isolation in production cementing; Sistematica para obtencao de isolamentos nas cimentacoes de producao

    Energy Technology Data Exchange (ETDEWEB)

    Lepeleire, Rubens de; Miletto, Antonio Luis Pires [PETROBRAS, XX (Brazil). Distrito de Perfuracao do Sudeste. Div. Tecnica

    1989-12-31

    This paper presents the procedure used to identify the factors that influenced the appearance of low isolation indexes, 59,1% in production cementing operations in Sao Mateus, during the year of 1987, bringing about a cost increase of over US$200,000.00 in primary cementing corrections. The various stages for the obtainment of such an index are analysed and the modifications that were considered necessary are presented. These modifications raised the index to 91,3% thus reducing correction costs to less than US$150,000.00 in 1988. From January to July, 1989, we able to obtain 98,9% of the isolations and an accumulated cementing correction cost of less than US$20,000.00. (author) 3 refs., 6 figs., 4 tabs.

  8. Isolation, characterization and formulation properties of a new plant gum obtained from Cissus refescence

    Directory of Open Access Journals (Sweden)

    Emeje Martins

    2009-01-01

    Full Text Available This study elucidated the physical, thermal, sorption and functional properties of a gum obtained from the stem of Cissus refescence (CRG. Scanning electron microscopy (SEM, Particle size analysis, X-ray powder diffraction (XPRD, Thermo gravimetric analysis (TGA, Differential scanning colorimetry (DSC, Fourier transmittance infra red (FTIR, and Elemental analysis were used to characterize the gum sample. Tablets were prepared by incorporating an anti asthmatic drug; theophylline. In vitro drug release was carried out in simulated gastric and intestinal conditions. Effect of gum concentration on release kinetics was evaluated. CRG had a glass transition (Tg and melting peak of 233.5 and 270 o C respectively. This material showed a 10.59 % loss in weight at 195 o C. The sample had very strong peaks at approximately 14 o , 15 o , 23 o , 24 o , and 29 o 2θ degrees of 2-theta (θ in the X-Ray Powder Diffraction pattern. Elemental analysis showed that CRG contains 44.1, 7.1, 48.5, and 0.3% Carbon, Hydrogen, Oxygen and Nitrogen respectively. Release of theophylline under simulated biologic conditions varied between 2 to 12 hours depending on the concentration of the gum used in formulation. Drug release was found to be erosion-controlled initially (i.e. in SGF, but at later stage, it became swelling -controlled (i.e. in SIF. The results obtained in this study establish the fundamental characteristics of CRG. The matrices were pH sensitive and can potentially be used for intestinal drug delivery.

  9. 40 CFR 180.1205 - Beauveria bassiana ATCC #74040; exemption from the requirements of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Beauveria bassiana ATCC #74040... RESIDUES IN FOOD Exemptions From Tolerances § 180.1205 Beauveria bassiana ATCC #74040; exemption from the... the insecticide Beauveria bassiana (ATCC #74040) in or on all food commodities when applied or used...

  10. 40 CFR 180.1273 - Beauveria bassiana HF23; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Beauveria bassiana HF23; exemption... FOOD Exemptions From Tolerances § 180.1273 Beauveria bassiana HF23; exemption from the requirement of a tolerance. Residues of Beauveria bassiana HF23 are exempt from the requirement of a tolerance on all...

  11. 40 CFR 180.1146 - Beauveria bassiana Strain GHA; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Beauveria bassiana Strain GHA... RESIDUES IN FOOD Exemptions From Tolerances § 180.1146 Beauveria bassiana Strain GHA; exemption from the requirement of a tolerance. Beauveria bassiana Strain GHA is exempted from the requirement of a tolerance...

  12. Binding of isolated plant lectin by rhizobia during episodes of reduced gravity obtained by parabolic flight

    Science.gov (United States)

    Henry, R. L.; Green, P. D.; Wong, P. P.; Guikema, J. A.; Spooner, B. S. (Principal Investigator)

    1990-01-01

    Development of a legume root nodule is a complex process culminating in a plant/bacterial symbiosis possessing the capacity for biological dinitrogen fixation. Formation of root nodules is initiated by the binding and stabilization of rhizobia to plant root hairs, mediated in part by a receptor/ligand recognition system composed of lectins on the plant root surface and lectin-binding sites on the rhizobial cell surface. The dinitrogen fixation activity of these root nodules may be an important feature of enclosed, space-based life support systems, and may provide an ecological method to recycle nitrogen for amino acid production. However, the effects on nodule development of varied gravitational fields, or of root nutrient delivery hardware, remain unknown. We have investigated the effects of microgravity on root nodule formation, with preliminary experiments focused upon the receptor/ligand component. Microgravity, obtained during parabolic flight aboard NASA 930, has no apparent effect on the binding of purified lectin to rhizobia, a result that will facilitate forthcoming experiments using intact root tissues.

  13. Antimicrobial resistance trends among Escherichia coli isolates obtained from dairy cattle in the northeastern United States, 2004-2011.

    Science.gov (United States)

    Cummings, Kevin J; Aprea, Victor A; Altier, Craig

    2014-01-01

    Monitoring antimicrobial resistance trends among bacteria isolated from food animals and people is necessary to inform risk analyses and guide public policy regarding antimicrobial use. Our objectives were to describe the antimicrobial resistance status of Escherichia coli isolates from dairy cattle in the northeastern United States and to identify trends in resistance to selected antimicrobial agents over time. We collected data retrospectively for all bovine E. coli isolates that were obtained from samples submitted to Cornell University's Animal Health Diagnostic Center between January 1, 2004 and December 31, 2011. We investigated temporal trends in the prevalence of resistant E. coli for each antimicrobial agent using the Cochran-Armitage trend test. Antimicrobial susceptibility testing was performed on 3373 bovine E. coli isolates from clinical samples submitted during the study period. Overall resistance to each antimicrobial agent ranged from 2.7% (enrofloxacin) to 91.3% (oxytetracycline). There was evidence of a significantly decreasing trend in prevalence of resistance to several agents: chlortetracycline, florfenicol, neomycin, oxytetracycline, spectinomycin, and trimethoprim/sulfamethoxazole. However, a significantly increasing trend in prevalence of resistance to enrofloxacin was also evident. These results do not support the idea that current antimicrobial use practices on dairy operations are driving a general increase in the emergence and dissemination of drug-resistant E. coli in the region served by the laboratory. However, resistance to some drugs remained consistently high during the study period, and increasing resistance to enrofloxacin is a key area of concern.

  14. Spore production of Beauveria bassiana from agro-industrial residues

    Directory of Open Access Journals (Sweden)

    Herta Stutz Dalla Santa

    2005-06-01

    Full Text Available The purpose of this work was to produce Beauveria bassiana by Solid-State Fermentation using agro-industrial residues and optimizing the cultivation conditions. Refused potatoes, coffee husks and sugar-cane bagasse were tested. The blend of refused potatoes and sugar-cane bagasse (60-40% with particle size in the range of 0.8-2 mm was used in the fermentation experiments. In Erlenmeyer flasks the best spore production was achieved with the following conditions: incubation temperature 26º C; initial pH 6.0; inoculum concentration 10(7 spores.g-1.dw and initial moisture 75%. In the column type reactor using forced aeration under the optimized conditions, the maximum production (1.07x10(10spores.g-1.dw was obtained at the 10th day of fermentation. The respirometric analyses of the fermentation showed a strong correlation between fungal growth and spore production.O objetivo deste trabalho foi produzir Beauveria bassiana por fermentação no estado sólido em resíduos agro-industriais e otimizar as condições de cultivo. Batata-refugo, polpa de café e bagaço de cana de açúcar foram testados. A mistura de batata-refugo e de bagaço de cana de açúcar (60:40%, com granulometria de 2 a 0,8 mm foi escolhida como melhor substrato/suporte. Em frascos de Erlenmeyer a produção de esporos foi maior com as seguintes condições: pH 6,0; temperatura de incubação de 26º C; taxa de inóculo de 10(7 esporos.g-1 de matéria seca; e umidade inicial de 75%. Em bioreator do tipo coluna com aeração forçada, as condições otimizadas possibilitaram uma produção máxima de esporos no 10º dia de fermentação, obtendo-se 1,07x10(10 esporos.g-1 de matéria seca. A análise respirométrica desta fermentação permitiu correlacionar o desenvolvimento do fungo com a produção de esporos.

  15. Characterization and virulence of Beauveria bassiana associated with auger beetle (Sinoxylon anale) infesting allspice (Pimenta dioica).

    Science.gov (United States)

    Senthil Kumar, C M; Jacob, T K; Devasahayam, S; D'Silva, Sharon; Nandeesh, P G

    2016-09-01

    The incidence of auger beetle, Sinoxylon anale Lesne (Bostrichidae: Coleoptera), a destructive pest of cosmopolitan occurrence is reported for the first time on allspice trees, Pimenta dioica (L.) Merr. in Kerala, India. The insects bored through the basal region of fresh twigs resulting in dieback symptoms. Morphological characterization and sequencing of a partially amplified fragment of the mitochondrial CO1 gene (696bp) revealed the insect to be Sinoxylon anale. An entomopathogenic fungus was isolated from infected cadavers of S. anale that was identified as Beauveria bassiana (Bals.-Criv.) Vuill., sensu stricto (s.s.) (Ascomycota: Hypocreales) based on morphological and molecular studies. The partial sequences of the ITS, TUB, TEF and Bloc gene regions were sequenced. The fungus grew well in ambient room temperature conditions (28-32±2°C; 60-70% relative humidity) and the infection process on the insect was documented by scanning electron microscopy. Bioassay studies with the isolate indicated that the fungus was virulent against adult beetles as evidenced by the LC50 (3.6×10(6)conidia/ml) and ST50 values (6.8days at a dose of 1×10(7)conidia/ml and 5.8days at a dose of 1×10(8)conidia/ml, respectively). This is the first record of B. bassiana naturally infecting S. anale and the fungus holds promise to be developed as a mycoinsecticide.

  16. Focus on RNA isolation: obtaining RNA for microRNA (miRNA) expression profiling analyses of neural tissue

    Science.gov (United States)

    Wang, Wang-Xia; Rajeev, Bernard W.; Baldwin, Donald A.; Isett, R. Benjamin; Ren, Na; Stromberg, Arnold; Nelson, Peter T.

    2008-01-01

    MicroRNAs (miRNAs) are present in all known plant and animal tissues and appear to be somewhat concentrated in the mammalian nervous system. Many different miRNA expression profiling platforms have been described. However, relatively little research has been published to establish the importance of ‘upstream’ variables in RNA isolation for neural miRNA expression profiling. We tested whether apparent changes in miRNA expression profiles may be associated with tissue processing, RNA isolation techniques, or different cell types in the sample. RNA isolation was performed on a single brain sample using eight different RNA isolation methods, and results were correlated using a conventional miRNA microarray and then cross-referenced to Northern blots. Differing results were seen between samples obtained using different RNA isolation techniques and between microarray and Northern blot results. Another complication of miRNA microarrays is tissue-level heterogeneity of cellular composition. To investigate this phenomenon, miRNA expression profiles were determined and compared between highly-purified primary cerebral cortical cell preparations of rat primary E15–E18 neurons versus rat primary E15–E18 astrocytes. Finally, to assess the importance of dissecting human brain gray matter from subjacent white matter in cerebral cortical studies, miRNA expression profiles were compared between gray matter and immediately contiguous white matter. The results suggest that for microarray studies, cellular composition is important, and dissecting white matter from gray matter improves the specificity of the results. Based on these data, recommendations for miRNA expression profiling in neural tissues, and considerations worthy of further study, are discussed. PMID:18316046

  17. Antimicrobial resistance trends among Salmonella isolates obtained from dairy cattle in the northeastern United States, 2004-2011.

    Science.gov (United States)

    Cummings, Kevin J; Perkins, Gillian A; Khatibzadeh, Sarah M; Warnick, Lorin D; Altier, Craig

    2013-04-01

    Monitoring antimicrobial resistance trends among bacteria isolated from food animals and people is necessary to inform public policy regarding appropriate antimicrobial use. Our objectives were to describe the antimicrobial resistance status of Salmonella isolates from dairy cattle in the northeastern United States and to identify trends in resistance to various antimicrobial agents over time. Data were collected retrospectively for all bovine Salmonella isolates that were obtained from samples submitted to Cornell University's Animal Health Diagnostic Center between January 1, 2004 and December 31, 2011. Temporal trends in the prevalence of resistant Salmonella were investigated for each antimicrobial agent using the Cochran-Armitage trend test. Antimicrobial susceptibility testing was performed on 2745 bovine Salmonella isolates from clinical samples submitted during the study period. Overall resistance to each antimicrobial agent ranged from 0% (amikacin, ciprofloxacin, and nalidixic acid) to 72.0% (sulfadimethoxine). There was evidence of a significantly decreasing trend in prevalence of resistance to most agents: amoxicillin/clavulanic acid (AUG), ampicillin (AMP), cefoxitin (FOX), ceftiofur (TIO), ceftriaxone (AXO), chloramphenicol (CHL), chlortetracycline (CTET), florfenicol (FFN), kanamycin (KAN), neomycin (NEO), oxytetracycline (OXY), spectinomycin (SPE), streptomycin (STR), sulfadimethoxine (SDM), sulfisoxazole (FIS), and tetracycline (TET). Among the 265 isolates that were tested using the National Antimicrobial Resistance Monitoring System (NARMS) panel, the most common resistance patterns were pansusceptible (54.0%), AUG-AMP-FOX-TIO-AXO-CHL-KAN-STR-FIS-TET (18.1%), and AUG-AMP-FOX-TIO-AXO-CHL-STR-FIS-TET (12.1%). Increasing prevalence of S. enterica serovar Cerro over the course of the study period presumably had an impact on the observed resistance trends. Nevertheless, these results do not support the notion that the current level of antimicrobial

  18. Paracoccidioides brasilienses isolates obtained from patients with acute and chronic disease exhibit morphological differences after animal passage

    Directory of Open Access Journals (Sweden)

    SVIDZINSKI Terezinha Inez Estivalet

    1999-01-01

    Full Text Available The basis for virulence in Paracoccidioides brasiliensis is not completely understood. There is a consensus that the sequencial in vitro subcultivation of P. brasiliensis leads to loss of its pathogenicity, which can be reverted by reisolation from animal passage. Attention to morphological and biochemical properties that are regained or demonstrated after animal passage may provide new insights into factors related to the pathogenicity and virulence of P. brasiliensis. We evaluated morphological characters: the percentage of budding cells, number of buds by cell and the diameter of 100 mother cells of yeast-like cells of 30 P. brasiliensis isolates, before and after animal passage. The isolates were obtained from patients with different clinical forms of paracoccidioidomycosis (PCM: acute form (group A, n=15 and chronic form (group C, n=15. The measurement of the yeast cell sizes was carried out with the aid of an Olympus CBB microscope coupled with a micrometer disc. We measured the major transverse and longitudinal axes of 100 viable cells of each preparation. The percentage of budding cells as also the number of buds by cell was not influenced by animal passage, regardless of the source of the strain (acute or chronic groups. The size values of P. brasiliensis isolates from groups A and C, measured before the animal passage exhibited the same behavior. After animal passage, there was a statistically significant difference between the cell sizes of P. brasiliensis isolates recovered from testicles inoculated with strains from groups A and C. The maximum diameter of mother cells from group A isolates exhibited a size of 42.1mm in contrast with 32.9mm exhibited by mother cells from group C (p<0.05. The diameter of 1500 mother cells from group A isolates exhibited a medium size of 16.0mm (SD ± 4.0, a value significantly higher than the 14.1mm (SD = ± 3.3 exhibited by 1500 mother cells from group C isolates (p<0.05. Our results reinforce the

  19. Molecular Genetics of Beauveria bassiana Infection of Insects.

    Science.gov (United States)

    Ortiz-Urquiza, A; Keyhani, N O

    2016-01-01

    Research on the insect pathogenic filamentous fungus, Beauveria bassiana has witnessed significant growth in recent years from mainly physiological studies related to its insect biological control potential, to addressing fundamental questions regarding the underlying molecular mechanisms of fungal development and virulence. This has been in part due to a confluence of robust genetic tools and genomic resources for the fungus, and recognition of expanded ecological interactions with which the fungus engages. Beauveria bassiana is a broad host range insect pathogen that has the ability to form intimate symbiotic relationships with plants. Indeed, there is an increasing realization that the latter may be the predominant environmental interaction in which the fungus participates, and that insect parasitism may be an opportunist lifestyle evolved due to the carbon- and nitrogen-rich resources present in insect bodies. Here, we will review progress on the molecular genetics of B. bassiana, which has largely been directed toward identifying genetic pathways involved in stress response and virulence assumed to have practical applications in improving the insect control potential of the fungus. Important strides have also been made in understanding aspects of B. bassiana development. Finally, although increasingly apparent in a number of studies, there is a need for progressing beyond phenotypic mutant characterization to sufficiently investigate the molecular mechanisms underlying B. bassiana's unique and diverse lifestyles as saprophyte, insect pathogen, and plant mutualist.

  20. Effectiveness of Beauveria bassiana sensu lato strains for biological control against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in China.

    Science.gov (United States)

    Sun, Ming; Ren, Qiaoyun; Guan, Guiquan; Li, Yufeng; Han, Xueqing; Ma, Chao; Yin, Hong; Luo, Jianxun

    2013-10-01

    Owing to the need to combat the spread of acaricide-resistant ticks, the development of long-term biological control has become a hot topic for tick control. In this study, we investigated the pathogenicity of three Beauveria bassiana isolates on the engorged female Rhipicephalus (Boophilus) microplus ticks using different conidial concentrations. The results showed that B. bassiana B.bAT17 was highly pathogenic against engorged R. (B.) microplus females, resulting in lethal time (LT50 and LT90) of 7.14 and 9.33 days at a concentration of 10(9)conidia/ml. R. (B.) microplus females treated with B. bassiana B.bAT17 significantly reduced the amount of ovipositioning; and most ticks died before they could begin to oviposit. Proteases and chitinases were analyzed in order to establish a screening method for identification of high virulent strains. This study has confirmed the significant pathogenic effect of entomopathogenic fungi against engorged R. (B.) microplus females in China, and further studies on the efficiency of the fungus against ticks in the field are required.

  1. Structure and antioxidant activity of soy protein isolate-dextran conjugates obtained by TiO2 photocatalysis.

    Science.gov (United States)

    Jin, Bei; Zhou, Xiaosong; Li, Bing; Chen, Caiyan; Zhang, Xiaosa; Chen, Siqiao

    2015-01-01

    The aim of this study was to investigate the structural characteristics and antioxidant activities of soy protein isolate- (SPI-) dextran conjugates obtained by TiO2 photocatalysis treatment. Results revealed that the UV-vis absorption and the fluorescence intensity increased as the photocatalytic power increased (P photocatalysis. Moreover, significant changes of secondary structure occurred in SPI-dextran conjugates. The α-helix, β-sheet, β-turns, and random coil were changed from approximately 10.6%, 37.9%, 12.9%, and 38.6% to 3.8%, 10.4%, 17.7%, and 68.8%, respectively, after treatment at photocatalytic power of 1000 W. In addition, SPI-dextran conjugates obtained by TiO2 photocatalysis treatment exhibited high hydroxyl radical scavenging activity and possessed increased reducing power. All data indicated that TiO2 photocatalysis was an efficient method for promoting protein-polysaccharide copolymerisation.

  2. New Approach of Beauveria bassiana to Control the Red Palm Weevil (Coleoptera: Curculionidae) by Trapping Technique.

    Science.gov (United States)

    Hajjar, M J; Ajlan, A M; Al-Ahmad, M H

    2015-04-01

    This work is the first study to investigate the efficacy of the commercial formulation of Beauveria bassiana (Broadband) to control adults of red palm weevil (Rhynchophorus ferrugineus (Olivier)). This fungus could be applied as one of the biological tactics in controlling red palm weevil. Bioassay experiments for medium lethal concentrate and medium time to cause death of 50% of red palm weevil adults were carried out. The result showed that the LC50 of B. bassiana (Broadband) was 2.19×10(7) and 2.76×10(6) spores/ml at 9 and 23 d of treatment, respectively. The LT50 was 13.95 and 4.15 d for concentration of 1×10(7) and 1×10(8) spores/ml, respectively, whereas 1×10(9) spores/ml caused 100% mortality after 24 h. Additionally, a red palm weevil pheromone trap was designed to attract the adults to be contaminated with spores of Broadband, which was applied to the sackcloth fabric that coated the internal surfaces of the bucket trap. The mating behavior was studied to determine direct and indirect infection of the spores from male to female and vice versa. The results showed a high efficacy of Broadband suspension at 1×10(9) spores/ml; 40 ml of suspension at this concentration treated to cloth in a trap caused death of contaminated adults with B. bassiana spores directly and indirectly. The 100% mortality was obtained even after 13 d of traps treatment with 40 ml of the suspension at 1×10(9) spores/ml.

  3. Identification of lactic acid bacteria with bio-preservative potential isolated from contaminated avian blood obtained at the slaughterhouse

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    MV Zbrun

    2013-01-01

    Full Text Available Blood is a common by-product of the meat industry, which has several potential applications in the animal feed industry. However, since blood is highly susceptible to microbial spoilage, blood and its fractions are often not suitable ingredients for the feed industry. Biopreservation appears as an alternative for the improvement of blood's quality towards its use as an ingredient in foodstuff. The objective of this work was to isolate and identify Lactic Acid Bacteria (LAB in avian blood obtained from industrial slaughterhouses and evaluate their antimicrobial activity. Ninety-six LAB were isolated from avian blood and genotyped. Eleven Amplified rDNA Restriction Analysis groups were identified. Between two and five different species were detected in each blood sample (31 strains in all blood samples which were selected to study antagonistic activity. Twenty-eight of them produced antimicrobial compounds such as organic acids, 11 strains produced hydrogen peroxide (H2O2 and two released bacteriocin-like compounds. The latter, identified as Lactobacillus salivarius (DSPV 027SA and Enterococcus faecalis (DSPV 008SA, inhibited the growth of Escherichia coli, Pseudomonas aeruginosa and some serotypes of Salmonella spp. These two LAB strains would be candidates for potential application as a blood biopreservation system. This biotechnological tool is cheaper than others sanitation techniques and could reduce the risk of pathogens transmission thought food chain.

  4. Effect of replacement of corn starch by whey protein isolate in biodegradable film blends obtained by extrusion.

    Science.gov (United States)

    Azevedo, Viviane Machado; Borges, Soraia Vilela; Marconcini, José Manoel; Yoshida, Maria Irene; Neto, Alfredo Rodrigues Sena; Pereira, Tamara Coelho; Pereira, Camila Ferreira Gonçalves

    2017-02-10

    The aim of this study was to evaluate the effect of replacing corn starch by whey protein isolated (WPI) in biodegradable polymer blends developed by extrusion. X-ray diffraction showed the presence of a Vh-type crystalline arrangement. The films were homogeneous, indicating strong interfacial adhesion between the protein and the thermoplastic starch matrix (TPS) as observed in scanning electron microscopy. The addition of WPI on TPS matrix promoted an increase in the thermal stability of the materials. It was observed 58.5% decrease in the water vapor permeability. The effect of corn starch substitution by WPI on mechanical properties resulted in a more resistant and less flexible film when compared the TPS film. The addition of WPI caused greenish yellow color and less transparent films. The substitution of corn starch by WPI made it possible to obtain polymer blends with improved properties and represents an innovation for application as a packaging material.

  5. Regulation of a phenylalanine ammonia lyase (BbPAL) by calmodulin in response to environmental changes in the entomopathogenic fungus Beauveria bassiana.

    Science.gov (United States)

    Kim, Jiyoung; Park, Hyesung; Han, Jae-Gu; Oh, Junsang; Choi, Hyung-Kyoon; Kim, Seong Hwan; Sung, Gi-Ho

    2015-11-01

    Phenylalanine ammonia lyase (PAL, E.C. 4.3.1.5) catalyses the deamination of L -phenylalanine to trans-cinnamic acid and ammonia, facilitating a critical step in the phenylpropanoid pathway that produces a variety of secondary metabolites. In this study, we isolated BbPAL gene in the entomopathogenic fungus Beauveria bassiana. According to multiple sequence alignment, homology modelling and in vitro PAL activity, we demonstrated that BbPAL acts as a typical PAL enzyme in B. bassiana. BbPAL interacted with calmodulin (CaM) in vitro and in vivo, indicating that BbPAL is a novel CaM-binding protein. The functional role of CaM in BbPAL action was to negatively regulate the BbPAL activity in B. bassiana. High-performance liquid chromatography analysis revealed that L -phenylalanine was reduced and trans-cinnamic acid was increased in response to the CaM inhibitor W-7. Dark conditions suppressed BbPAL activity in B. bassiana, compared with light. In addition, heat and cold stresses inhibited BbPAL activity in B. bassiana. Interestingly, these negative effects of BbPAL activity by dark, heat and cold conditions were recovered by W-7 treatment, suggesting that the inhibitory mechanism is mediated through stimulation of CaM activity. Therefore, this work suggests that BbPAL plays a role in the phenylpropanoid pathway mediated by environmental stimuli via the CaM signalling pathway.

  6. Toxicity of entomopathogenic fungi, Beauveria bassiana and Lecanicillium muscarium against a field-collected strain of the German cockroach Blattella germanica (L.) (Dictyoptera: Blattellidae).

    Science.gov (United States)

    Davari, B; Limoee, M; Khodavaisy, S; Zamini, G; Izadi, S

    2015-09-01

    The German cockroach, Blattella germanica (L.) has been recognized as a serious health problem throughout the world. Control failures due to insecticide resistance and chemical contamination of environment have led some researchers focus on the other alternative strategy controls. Microbial insecticides such as those containing entomo pathogenic fungi could be of high significance. Lecanicillium muscarium and Beauveria bassiana grow naturally in soils throughout the world and act as a parasite on various arthropod species, causing white muscardine disease. Thus, these two species could be considered as entomopathogenic fungi. The current study conducted to evaluate the toxicity of Beauveria bassiana and Lecanicillium muscarium against German cockroach, Blattella germanica. Conidial formulations of L. muscarium (PTCC 5184) and B. bassiana (PTCC5197) were prepared in aqueous suspensions with Tween 20. Bioassays were performed using two methods including submersion of cockroaches in conidial suspension and baiting. Data were analyzed by Probit program and LC50 and LC90 were estimated. The obtained results indicated that both fungi species were toxic against German cockroach however; Beauveria bassiana was significantly 4.8 fold more toxic than L. muscarium against German cockroach using submersion method.

  7. Detection of sul1, sul2 and sul3 in sulphonamide resistant Escherichia coli isolates obtained from healthy humans, pork and pigs in Denmark.

    Science.gov (United States)

    Hammerum, Anette M; Sandvang, Dorthe; Andersen, Sigrid R; Seyfarth, Anne Mette; Porsbo, Lone Jannok; Frimodt-Møller, Niels; Heuer, Ole E

    2006-02-01

    The occurrence of sulphonamide resistance was investigated in 998 Escherichia coli isolates, obtained from pig faeces collected at slaughter, Danish pork collected at retail outlets and from faeces from healthy persons in Denmark. In total 18% (n=35), 20% (n=38) and 26% (n=161) of the E. coli isolates obtained from humans, pork and pigs, respectively, were resistant to sulphonamide. All sulphonamide resistant E. coli isolates were investigated for the presence of sul1, sul2, sul3 and intI1 genes by PCR. The sul1 gene was detected in 40% (n=14), 29% (n=11) and 55% (n=88) of the sulphonamide resistant isolates from humans, pork and pigs, respectively. The sul2 gene was detected in 80% (n=28), 76% (n=29) and 50% (n=81) of isolates from humans, pork and pigs, respectively. None of the human isolates were PCR-positive for sul3, whereas sul3 was present in 5% of the pork isolates and 11% of the pig isolates. Of the 113 sul1 positive isolates, 97 carried the integron-associated integrase gene intI1. All 20 sul3 positive isolates were positive for intI1, and in 12 of these isolates sul3 was the only sulphonamide resistance gene detected. The origin of sul1 and sul2 found in isolates from healthy humans is speculative, but their spread from pigs to humans via the food chain is possible.

  8. Structure and Antioxidant Activity of Soy Protein Isolate-Dextran Conjugates Obtained by TiO2 Photocatalysis

    Directory of Open Access Journals (Sweden)

    Bei Jin

    2015-01-01

    Full Text Available The aim of this study was to investigate the structural characteristics and antioxidant activities of soy protein isolate- (SPI- dextran conjugates obtained by TiO2 photocatalysis treatment. Results revealed that the UV-vis absorption and the fluorescence intensity increased as the photocatalytic power increased (P<0.05. Higher photocatalytic power could promote the extent of glycation and the formation of high molecular weight SPI-dextran conjugates, which were evidenced by free amino group content and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE analysis. The Fourier transform infrared (FT-IR spectra suggested that the amide I, II, and III bands of SPI were altered by the glycation induced by TiO2 photocatalysis. Moreover, significant changes of secondary structure occurred in SPI-dextran conjugates. The α-helix, β-sheet, β-turns, and random coil were changed from approximately 10.6%, 37.9%, 12.9%, and 38.6% to 3.8%, 10.4%, 17.7%, and 68.8%, respectively, after treatment at photocatalytic power of 1000 W. In addition, SPI-dextran conjugates obtained by TiO2 photocatalysis treatment exhibited high hydroxyl radical scavenging activity and possessed increased reducing power. All data indicated that TiO2 photocatalysis was an efficient method for promoting protein-polysaccharide copolymerisation.

  9. Isolation and Identification of Aerobic Bacteria Carrying Tetracycline and Sulfonamide Resistance Genes Obtained from a Meat Processing Plant.

    Science.gov (United States)

    Li, Lili; Ye, Lei; Zhang, Sen; Meng, Hecheng

    2016-06-01

    Microbial contamination in food-processing plants can play a fundamental role in food quality and safety. The purpose of this study was to investigate aerobic bacteria carrying tetracycline and sulfonamide resistance genes from a meat processing plant as possible sources of meat contamination. One hundred swab samples from surfaces of conveyor belts, meat slicers, meat knives, benches, plastic trays, gloves, and aprons were analyzed. A total of 168 isolates belonging to 10 genera were obtained, including Pseudomonas sp. (n = 35), Acinetobacter sp. (n = 30), Aeromonas sp. (n = 20), Myroides sp. (n = 15), Serratia sp. (n = 15), Staphylococcus sp. (n = 14), Enterobacter sp. (n = 11), Escherichia coli (n = 10), Lactococcus sp. (n = 10), and Klebsiella sp. (n = 8). Of the 168 isolates investigated, 60.7% showed resistance to tetracycline and 57.7% to trimethoprim/sulfamethoxazole. The tetracycline resistance genes tetL, tetA, tetB, tetC, tetE, tetM, tetS, tetK, and tetX were found in the frequency of 7.7%, 6.0%, 4.8%, 4.8%, 3.6%, 3.6%, 3.6%, 1.2%, and 0.6%, respectively. Sulfonamide resistance genes sul1 and sul2 were observed in the frequency of 17.9% and 38.1%, respectively. The tetracycline resistance genes tetX was first found in Myroides sp. This investigation demonstrated that food contact surfaces in a meat processing plant may be sources of contamination of aerobic bacteria carrying tetracycline and sulfonamide antibiotic resistance genes. © 2016 Institute of Food Technologists®

  10. Evaluation of effectiveness of entomopathogenic fungi Beauveria bassiana using a standard laboratory bioassay

    OpenAIRE

    2011-01-01

    In laboratory bioassays, the efficacy of the entomopathogenic fungus Beauveria bassiana against the yellow mealworm (Tenebrio molitor) was tested under various temperature conditions. Six different strains of fungus B. bassiana was investigated. The evaluation was based on vitality bioassays including germination and growth index assessment and the bioassay of virulence based on target organism T. molitor was also assessed growth and yield of conidia different strains of fungus B. bassiana on...

  11. Roseomonas tokyonensis sp. nov. isolated from a biofilm sample obtained from a cooling tower in Tokyo, Japan.

    Science.gov (United States)

    Furuhata, Katsunori; Ishizaki, Naoto; Edagawa, Akiko; Fukuyama, Masafumi

    2013-01-01

    Strain K-20(T), a Gram-negative, nonmotile, nonspore-forming and strictly aerobic coccobacillus, which produces a pale pink pigment (R2A agar medium, 30℃, seven days) was isolated from a sample of biofilm obtained from a cooling tower in Tokyo, Japan. A phylogenetic analysis of the 16S rRNA partial gene sequences (1,439 bp) showed that the strain (accession number: AB297501) was related to Roseomonas frigidaquae CW67(T) and Roseomonas stagni HS-69(T) with 97.4% and 96.9% sequence similarity, respectively. Strain K-20(T) formed a distinct cluster with Roseomonas frigidaquae CW67(T) in the phylogenetic tree at a high bootstrap value (93%); however, distance was recognized between the strains. In addition, the DNA-DNA hybridization level between strain K-20(T) and Roseomonas frigidaquae JCM 15073(T) was 33%. The taxonomic data indicate that K-20(T) (=JCM 14634(T) =KCTC 32152(T)) should be classified in the genus Roseomonas as the type strain of a novel species, Roseomonas tokyonensis sp. nov.

  12. Ultrastructural study of the mycelial phase of clinical isolates of Sporothrix schenckii obtained from feline, canine and human cases of sporotrichosis

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    Isabel Martins Madrid

    2011-09-01

    Full Text Available Using transmission electron microscopy, we studied the presence of melanin and cell wall thickness of clinical isolates of Sporothrix schenckii obtained from cats, dogs and humans as compared to reference strains. We detected differences regarding presence of the melanin among the clinical isolates of S. schenckii and a correlation between presence of melanin and cell wall thickness.

  13. Compatibility of the insect pathogenic fungus Beauveria bassiana with neem against sweetpotato whitefly, Bemisia tabaci, on eggplant

    Science.gov (United States)

    A study on the compatibility of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) with neem was conducted against sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), on eggplant. Initially, three concentrations of B. bassiana (106, 1...

  14. The use of aggregation pheromone to enhance dissemination of Beauveria bassiana for the control of the banana weevil in Uganda

    NARCIS (Netherlands)

    Tinzaara, W.; Gold, C.S.; Dicke, M.; Huis, van A.; Nankinga, C.M.; Kagezi, G.H.; Ragama, P.E.

    2007-01-01

    Candidate strains of Beauveria bassiana were identified for use in integrated pest management of the banana weevil Cosmopolites sordidus. Horizontal field transmission of B. bassiana between banana weevils using different delivery systems, including aggregation pheromones, was investigated. We obser

  15. Molecular characterization and phylogeny of Shiga toxin-producing Escherichia coli isolates obtained from two Dutch regions using whole genome sequencing.

    Science.gov (United States)

    Ferdous, M; Friedrich, A W; Grundmann, H; de Boer, R F; Croughs, P D; Islam, M A; Kluytmans-van den Bergh, M F Q; Kooistra-Smid, A M D; Rossen, J W A

    2016-07-01

    Shiga toxin-producing Escherichia coli (STEC) is one of the major causes of human gastrointestinal disease and has been implicated in sporadic cases and outbreaks of diarrhoea, haemorrhagic colitis and haemolytic uremic syndrome worldwide. In this study, we determined the molecular characteristics and phylogenetic relationship of STEC isolates, and their genetic diversity was compared to that of other E. coli populations. Whole genome sequencing was performed on 132 clinical STEC isolates obtained from the faeces of 129 Dutch patients with gastrointestinal complaints. STEC isolates of this study belonged to 44 different sequence types (STs), 42 serogenotypes and 14 stx subtype combinations. Antibiotic resistance genes were more frequently present in stx1-positive isolates compared to stx2 and stx1 + stx2-positive isolates. The iha, mchB, mchC, mchF, subA, ireA, senB, saa and sigA genes were significantly more frequently present in eae-negative than in eae-positive STEC isolates. Presence of virulence genes encoding type III secretion proteins and adhesins was associated with isolates obtained from patients with bloody diarrhoea. Core genome phylogenetic analysis showed that isolates clustered according to their ST or serogenotypes irrespective of stx subtypes. Isolates obtained from patients with bloody diarrhoea were from diverse phylogenetic backgrounds. Some STEC isolates shared common ancestors with non-STEC isolates. Whole genome sequencing is a powerful tool for clinical microbiology, allowing high-resolution molecular typing, population structure analysis and detailed molecular characterization of strains. STEC isolates of a substantial genetic diversity and of distinct phylogenetic groups were observed in this study.

  16. Evaluation of kinetic parameters of chitinases produced by Beauveria bassiana (Bals. Vuill. / Avaliação de parâmetros cinéticos de quitinases produzidas por Beauveria bassiana (Bals. Vuill.

    Directory of Open Access Journals (Sweden)

    Cristiane Mita

    2008-07-01

    Full Text Available Entomopathogenic fungus Beauveria bassiana is currently used as a biocontrol agent for agricultural pests. The infection process involves extracellular enzymes such as proteases and chitinases that degrade the cuticle of the insects. The objective of this work was to evaluate kinetic parameters of pH, temperature, ionic concentration and time of reaction on chitinases activity. The fungus B. bassiana CG432 was cultivated on coffee berry borer Hypothenemus hampei (Ferrari and the conidia grown on insect were used to prepare the inoculum containing 108conídia/mL. These conidia were inoculated at 1% (v/v in culture liquid medium containing D-glucose (10g, yeast extract (5g, NaNO3 (1,58g, Na2HPO4.7H2O (1,05g, KCl (1g, MgSO4.7H2O (0,6g and KH2PO4 (0,36g per liter. The cultivation was carried at 28°C and 180rpm during 5 days. Culture fluid was obtained by filtration and centrifugation at 8.000g, and the chitinases were isolated and concentrated by ultrafiltration using 10 and 100kDa cut off membranes under nitrogen pressure. Chitinase activity was detected and quantified using N-acetylglucosamine released by hydrolysis of colloidal chitin at 40 to 60ºC, at 50, 100 and 200 mM ionic concentrations of buffers sodium acetate (pH 4.0 to 6.0; sodium phosphate (pH 6.0 to 8.0; and Glycine-NaOH (pH 8.0 to 10.0 during 60 minutes. Maximum chitinase activity was at 45ºC and pH 5.5, and was also high at pH 6.0 and pH 8.5 using 50mM buffer. The chitinase activity increased and was stable during an hour at optimum conditions of the reaction, shown the stable nature of this enzyme.Beauveria bassiana é um fungo entomopatogênico utilizado no controle biológico de insetos-praga que infestam produtos agrícolas. O mecanismo de infecção envolve a produção de enzimas extracelulares, como proteases e quitinases que degradam a cutícula dos insetos. O objetivo deste trabalho foi avaliar parâmetros cinéticos de pH, temperatura, concentração iônica e tempo de

  17. Genetic diversity demonstrated by pulsed field gel electrophoresis of Salmonella enterica isolates obtained from diverse sources in Mexico

    Science.gov (United States)

    This study was conducted to determine the genetic diversity of Salmonella isolates recovered from a variety of sources using pulsed-field gel electrophoresis (PFGE) to assess their possible relatedness. Salmonella was isolated from ca. 52% of samples from a pepper var. Bell production system. A to...

  18. Multiple-stress tolerance of ionizing radiation-resistant bacterial isolates obtained from various habitats: correlation between stresses.

    Science.gov (United States)

    Shukla, Manish; Chaturvedi, Ruchi; Tamhane, Dhruti; Vyas, Pranav; Archana, G; Apte, Shree; Bandekar, J; Desai, Anjana

    2007-02-01

    Isolation of five ionizing radiation (IR)-resistant bacteria by screening of isolates from various habitats classified as common and stressed is reported. IR-resistant isolates exhibited varying degrees of resistance to gamma-radiation and were classified as highly and moderately radiation resistant. Resistance to ultraviolet (UV) radiation correlated well with gamma-radiation resistance, whereas a comparable desiccation resistance for all the highly and moderately radiation-resistant isolates was observed. However, salt tolerance failed to correlate with IR resistance, indicating a divergent evolution of the salt tolerance and radiation resistance. Characterization of isolates by the amplified rDNA restriction analysis profiling attested to the clustering of these isolates with their stress phenotype. 16S rRNA gene-based analysis of the isolates showed that the bacteria with similar-resistance physiologies clustered together and belonged to related genera. Hydrogen peroxide resistance and mitomycin survival patterns of the isolates indicated the roles of oxidative-stress tolerance in desiccation survival and recombination repair in higher radiation resistance, respectively.

  19. Transmission of Beauveria bassiana from male to female Aedes aegypti mosquitoes

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    Reyes-Villanueva Filiberto

    2011-02-01

    Full Text Available Abstract Background Resistance to chemical insecticides plus high morbidity rates have lead to rising interest in fungi as candidates for biocontrol agents of mosquito vectors. In most studies fungal infections have been induced by exposure of mosquitoes to various surfaces treated with conidia. In the present study eight Mexican strains of Beauveria bassiana were assessed against Aedes aegypti by direct exposure of females to 6 × 108 conidia ml -1 on a filter paper, afterwards, the transmission of the least and most virulent isolates was evaluated by mating behavior from virgin, fungus-contaminated male to females, to examine this ethological pattern as a new approach to deliver conidia against the dengue vector. Methods In an exposure chamber with a filter paper impregnated with 6 × 108 conidia ml -1 of the least and most virulent strains of B. bassiana, 6-8 day old males of A. aegypti were exposed for 48 hours, and then transferred individually (each one was a replicate to another chamber and confined with twenty healthy females of the same age. Clean males were used in controls. Survival, infection by true mating (insemination or by mating attempts (no insemination and fecundity were daily registered until the death of last female. Data analysis was conducted with proc glm for unbalanced experiments and means were separated with the Ryan test with SAS. Results All strains were highly virulent with LT50 ranging from 2.70 (± 0.29 to 5.33 (± 0.53 days. However the most (Bb-CBG2 and least virulent (Bb-CBG4 isolates were also transmitted by mating behavior; both killed 78-90% of females in 15 days after being confined with males that had previously been exposed for 48 hours to fungi. Of these mortality rates, 23 and 38% respectively, were infections acquired by copulations where insemination occurred. The LT50 for sexually-infected females were 7.92 (± 0.46 and 8.82 (± 0.45 days for both strains, while the one in control was 13.92 (± 0

  20. Transmission of Beauveria bassiana from male to female Aedes aegypti mosquitoes.

    Science.gov (United States)

    García-Munguía, Alberto M; Garza-Hernández, Javier A; Rebollar-Tellez, Eduardo A; Rodríguez-Pérez, Mario A; Reyes-Villanueva, Filiberto

    2011-02-26

    Resistance to chemical insecticides plus high morbidity rates have lead to rising interest in fungi as candidates for biocontrol agents of mosquito vectors. In most studies fungal infections have been induced by exposure of mosquitoes to various surfaces treated with conidia. In the present study eight Mexican strains of Beauveria bassiana were assessed against Aedes aegypti by direct exposure of females to 6 × 10(8) conidia ml (-1) on a filter paper, afterwards, the transmission of the least and most virulent isolates was evaluated by mating behavior from virgin, fungus-contaminated male to females, to examine this ethological pattern as a new approach to deliver conidia against the dengue vector. In an exposure chamber with a filter paper impregnated with 6 × 10(8) conidia ml (-1) of the least and most virulent strains of B. bassiana, 6-8 day old males of A. aegypti were exposed for 48 hours, and then transferred individually (each one was a replicate) to another chamber and confined with twenty healthy females of the same age. Clean males were used in controls. Survival, infection by true mating (insemination) or by mating attempts (no insemination) and fecundity were daily registered until the death of last female. Data analysis was conducted with proc glm for unbalanced experiments and means were separated with the Ryan test with SAS. All strains were highly virulent with LT(50) ranging from 2.70 (± 0.29) to 5.33 (± 0.53) days. However the most (Bb-CBG2) and least virulent (Bb-CBG4) isolates were also transmitted by mating behavior; both killed 78-90% of females in 15 days after being confined with males that had previously been exposed for 48 hours to fungi. Of these mortality rates, 23 and 38% respectively, were infections acquired by copulations where insemination occurred. The LT(50) for sexually-infected females were 7.92 (± 0.46) and 8.82 (± 0.45) days for both strains, while the one in control was 13.92 (± 0.58). Likewise, fecundity decreased by

  1. Existence of entomopathogen fungi, Beauveria bassiana as an endophyte in cacao seedlings

    Directory of Open Access Journals (Sweden)

    Endang Sulistyowati

    2015-12-01

    Full Text Available Beauveria bassiana is one of the entomopathogen fungi which is known as biological control agent of cocoa pod borer and cocoa mirids (Helopeltis spp.. Because of its effectiveness in the fields is still not consistent, so we conduct a research with the objective to know the possibility of Beauveria bassiana to be established as a endophyte. Various fungal entomopathogens have already been reported as endophytes and the various methods used to inoculate the plants with B. bassiana were partially effective. The research has been conducted in laboratory of Plant Protection, Indonesian Coffee and Cocoa Research Institute by inoculating of cocoa seeds and cocoa nursery with B. bassiana suspension.  The trial was arranged  by randomized complete block design with a factorial arrangement. The factor were spore concentration of B. bassiana (0; 2; and 4 g/ 10 l and cocoa varieties (family of ICS 60, TSH858, and hybrid. The trial were use  four replications. The results showed that the fungal entomopathogen B. bassiana was established as an endophyte in cocoa seedling, both from cocoa seeds and nursery application. Percentage of existence of B. bassiana colonies as endophytes one month after seeds application were ICS 60 amounted to 93.3 % both on concentration treatments, while the families of TSH 858 by 80 % and 86.67 % respectively in 2 g and 4 g per 10 l of B. bassiana spores concentration treament.. The lowest percentage was in hybrids, which amounted to 66.67% and 50%. B. bassiana colonies was exixtence as an endophyte in culture from root, stem and leaves of cocoa seedling up to 5 months post inoculation. While the application on nursery by soil drenshing, leaf spraying, and stem injection , it was known that B. bassiana colonies were found in the tissues of leaves, stems, and roots until two months after application. Colonies of B. bassiana as endophytes still exsist until six weeks after nursery was planted in the field. 

  2. Genetic Relatedness of Salmonella Serovars Isolated from Catfish (Clarias gariepinus) and Tilapia (Tilapia mossambica) Obtained from Wet Markets and Ponds in Penang, Malaysia.

    Science.gov (United States)

    Budiati, Titik; Rusul, Gulam; Wan-Abdullah, Wan Nadiah; Chuah, Li-Oon; Ahmad, Rosma; Thong, Kwai Lin

    2016-04-01

    A total of 43 Salmonella enterica isolates belonging to different serovars (Salmonella Albany, Salmonella Agona, Salmonella Corvallis, Salmonella Stanley, Salmonella Typhimurium, Salmonella Mikawasima, and Salmonella Bovismorbificans) were isolated from catfish (Clarias gariepinus) and tilapia (Tilapia mossambica) obtained from nine wet markets and eight ponds in Penang, Malaysia. Thirteen, 19, and 11 isolates were isolated from 9 of 32 catfish, 14 of 32 tilapia, and 11 of 44 water samples, respectively. Fish reared in ponds were fed chicken offal, spoiled eggs, and commercial fish feed. The genetic relatedness of these Salmonella isolates was determined by random amplified polymorphic DNA PCR (RAPD-PCR) using primer OPC2, repetitive extragenic palindromic PCR (REP-PCR), and pulsed-field gel electrophoresis (PFGE). Composite analysis of the RAPD-PCR, REP-PCR, and PFGE results showed that the Salmonella serovars could be differentiated into six clusters and 15 singletons. RAPD-PCR differentiated the Salmonella isolates into 11 clusters and 10 singletons, while REP-PCR differentiated them into 4 clusters and 1 singleton. PFGE differentiated the Salmonella isolates into seven clusters and seven singletons. The close genetic relationship of Salmonella isolates from catfish or tilapia obtained from different ponds, irrespective of the type of feed given, may be caused by several factors, such as the quality of the water, density of fish, and size of ponds.

  3. Isolation and Characterization of Mobile Genetic Elements from Microbial Assemblages Obtained from the Field Research Center Site

    Energy Technology Data Exchange (ETDEWEB)

    Patricia Sobecky; Cassie Hodges; Kerri Lafferty; Mike Humphreys; Melanie Raimondo; Kristin Tuttle; Tamar Barkay

    2004-03-17

    Considerable knowledge has been gained from the intensive study of a relatively limited group of bacterial plasmids. Recent efforts have begun to focus on the characterization of, at the molecular level, plasmid populations and associated mobile genetic elements (e.g., transposons, integrons) occurring in a wider range of aquatic and terrestrial habitats. Surprisingly, however, little information is available regarding the incidence and distribution of mobile genetic elements extant in contaminated subsurface environments. Such studies will provide greater knowledge on the ecology of plasmids and their contributions to the genetic plasticity (and adaptation) of naturally occurring subsurface microbial communities. We requested soil cores from the DOE NABIR Field Research Center (FRC) located on the Oak Ridge Reservation. The cores, received in February 2003, were sampled from four areas on the Oak Ridge Site: Area 1, Area 2, Area 3 (representing contaminated subsurface locales) and the background reference sites. The average core length (24 in) was subdivided into three profiles and soil pH and moisture content were determined. Uranium concentration was also determined in bulk samples. Replicate aliquots were fixed for total cell counts and for bacterial isolation. Four different isolation media were used to culture aerobic and facultative microbes from these four study areas. Colony forming units ranged from a minimum of 100 per gram soil to a maximum of 10,000 irrespective of media composition used. The vast majority of cultured subsurface isolates were gram-positive isolates and plasmid characterization was conducted per methods routinely used in the Sobecky laboratory. The percentage of plasmid incidence ranged from 10% to 60% of all isolates tested. This frequency appears to be somewhat higher than the incidence of plasmids we have observed in other habitats and we are increasing the number of isolates screened to confirm this observation. We are also

  4. Multiple heavy metal removal using an entomopathogenic fungi Beauveria bassiana.

    Science.gov (United States)

    Gola, Deepak; Dey, Priyadarshini; Bhattacharya, Arghya; Mishra, Abhishek; Malik, Anushree; Namburath, Maneesh; Ahammad, Shaikh Ziauddin

    2016-10-01

    Towards the development of a potential remediation technology for multiple heavy metals [Zn(II), Cu(II), Cd(II), Cr(VI) and Ni(II)] from contaminated water, present study examined the growth kinetics and heavy metal removal ability of Beauveria bassiana in individual and multi metals. The specific growth rate of B. bassiana varied from 0.025h(-1) to 0.039h(-1) in presence of individual/multi heavy metals. FTIR analysis indicated the involvement of different surface functional groups in biosorption of different metals, while cellular changes in fungus was reflected by various microscopic (SEM, AFM and TEM) analysis. TEM studies proved removal of heavy metals via sorption and accumulation processes, whereas AFM studies revealed increase in cell surface roughness in fungal cells exposed to heavy metals. Present study delivers first report on the mechanism of bioremediation of heavy metals when present individually as well as multi metal mixture by entomopathogenic fungi.

  5. Comparison of host cell invasion and proliferation among Neospora caninum isolates obtained from oocysts and from clinical cases of naturally infected dogs.

    Science.gov (United States)

    Dellarupe, A; Regidor-Cerrillo, J; Jiménez-Ruiz, E; Schares, G; Unzaga, J M; Venturini, M C; Ortega-Mora, L M

    2014-10-01

    In a previous study we have shown that the in vitro invasion rate (IR) and tachyzoite yield (TY) are associated with the virulence phenotypes of Neospora caninum isolates of bovine origin. In addition, we recently observed marked differences in virulence when canine isolates were compared in a pregnant BALB/c mouse model. In this study, we investigated whether invasion and proliferation capacities could be used as virulence-related N. caninum phenotypic traits. Of the isolates compared in mice, four canine isolates obtained from oocysts (Nc-Ger2, Nc-Ger3, Nc-Ger-6, Nc-6 Arg) had shown a low-moderate virulence, and two further isolates obtained from dogs with neurological signs (Nc-Bahia, Nc-Liv) were highly virulent. The IR for each isolate was determined by a plaque assay and the counting of immunofluorescence-labeled parasitophorous vacuoles at 3 days post-inoculation (p.i.). The TY was determined by the quantification of tachyzoites at 56 h p.i. by real-time PCR. Most of the canine isolates showed similar IR values under controlled invasion conditions for 4h and 72 h p.i., indicating a limited time period for invasion similar to that observed for bovine isolates. The Nc-Ger3, Nc-Bahia, and Nc-Liv isolates showed a significantly higher IR and TY than the Nc-Ger2 and Nc-Ger6 isolates (P0.885, P0.8117, P<0.058) in mice. These results demonstrate the importance both the invasive and proliferative capacities have on the virulence of canine N. caninum isolates.

  6. Uncertainty and Sensitivity Analysis Results Obtained in the 1996 Performance Assessment for the Waste Isolation Pilot Plant

    Energy Technology Data Exchange (ETDEWEB)

    Bean, J.E.; Berglund, J.W.; Davis, F.J.; Economy, K.; Garner, J.W.; Helton, J.C.; Johnson, J.D.; MacKinnon, R.J.; Miller, J.; O' Brien, D.G.; Ramsey, J.L.; Schreiber, J.D.; Shinta, A.; Smith, L.N.; Stockman, C.; Stoelzel, D.M.; Vaughn, P.

    1998-09-01

    The Waste Isolation Pilot Plant (WPP) is located in southeastern New Mexico and is being developed by the U.S. Department of Energy (DOE) for the geologic (deep underground) disposal of transuranic (TRU) waste. A detailed performance assessment (PA) for the WIPP was carried out in 1996 and supports an application by the DOE to the U.S. Environmental Protection Agency (EPA) for the certification of the WIPP for the disposal of TRU waste. The 1996 WIPP PA uses a computational structure that maintains a separation between stochastic (i.e., aleatory) and subjective (i.e., epistemic) uncertainty, with stochastic uncertainty arising from the many possible disruptions that could occur over the 10,000 yr regulatory period that applies to the WIPP and subjective uncertainty arising from the imprecision with which many of the quantities required in the PA are known. Important parts of this structure are (1) the use of Latin hypercube sampling to incorporate the effects of subjective uncertainty, (2) the use of Monte Carlo (i.e., random) sampling to incorporate the effects of stochastic uncertainty, and (3) the efficient use of the necessarily limited number of mechanistic calculations that can be performed to support the analysis. The use of Latin hypercube sampling generates a mapping from imprecisely known analysis inputs to analysis outcomes of interest that provides both a display of the uncertainty in analysis outcomes (i.e., uncertainty analysis) and a basis for investigating the effects of individual inputs on these outcomes (i.e., sensitivity analysis). The sensitivity analysis procedures used in the PA include examination of scatterplots, stepwise regression analysis, and partial correlation analysis. Uncertainty and sensitivity analysis results obtained as part of the 1996 WIPP PA are presented and discussed. Specific topics considered include two phase flow in the vicinity of the repository, radionuclide release from the repository, fluid flow and radionuclide

  7. Presence of superantigen genes and antimicrobial resistance in Staphylococcus isolates obtained from the uteri of dairy cows with clinical endometritis.

    Science.gov (United States)

    Zhao, J-L; Ding, Y-X; Zhao, H-X; He, X-L; Li, P-F; Li, Z-F; Guan, H; Guo, X

    2014-10-11

    Clinical endometritis is an important disease of dairy cattle and results in decreased reproductive performance. This disease is caused by contamination of the uterus with a broad spectrum of microorganisms after calving. In this study, staphylococcal isolates from the uterus of dairy cows with clinical endometritis were tested for their distribution of superantigen (SAg) genes and antimicrobial resistance. Between the 127 staphylococcal isolates collected in this study, 10 species were identified. The predominant strain identified was Staphylococcus aureus (n=53), followed by Staphylococcus saprophyticus (n=38) and Staphylococcus chromogenes (n=22). PCR analysis demonstrated that most isolates (63.0 per cent) harboured at least one SAg gene. The most commonly observed SAg gene and genotype was selj (38.6 per cent) and sec-selj-seln (24.0 per cent), respectively. Most isolates were resistant to penicillin (79.5 per cent), ampicillin (71.7 per cent), erythromycin (56.7 per cent), and tetracycline (52.0 per cent). PCR analysis demonstrated that the antimicrobial resistance determinants ermA, ermB, ermC, tetK, tetM and blaZ were detected in 0 per cent, 44.4 per cent, 51.4 per cent, 68.2 per cent, 13.6 per cent and 86.1 per cent of the erythromycin, tetracycline and β-lactam resistant isolates, respectively. There were 22 (17.3 per cent of all isolates) coagulase-negative staphylococci shown to be methicillin resistant. In the methicillin-resistant isolates, significant resistances to ampicillin, erythromycin and penicillin were observed (P<0.01). The results of this study demonstrate that staphylococci recovered from dairy cows with clinical endometritis contain an extensive and complex prevalence of SAg genes. Significant resistances to antibiotics were also seen, highlighting the need for the rational appliance of antibiotics in veterinary medicine.

  8. Syk-Mediated Suppression of Inflammatory Responses by Cordyceps bassiana.

    Science.gov (United States)

    Yang, Woo Seok; Nam, Gyeong Sug; Kim, Mi-Yeon; Cho, Jae Youl

    2017-01-01

    The fruit body of artificially cultivated Cordyceps bassiana has been reported to exhibit anti-inflammatory and anticancer activities. Although it has been suggested that the fruit body has neutraceutic and pharmaceutic biomaterial potential, the exact anti-inflammatory molecular mechanism has not been fully elucidated. In this study, we demonstrated the immunopharmacologic activity of Cordyceps bassiana under in vitro conditions and investigated its anti-inflammatory mechanism. Water extract (Cm-WE) of the fruit body of artificially cultivated Cordyceps bassiana without polysaccharide fractions reduced the expression of the proinflammatory genes cyclooxygenase (COX)-2, interleukin (IL)-12, and inducible nitric oxide synthase (iNOS) and promoted the expression of the anti-inflammatory gene IL-10 in lipopolysaccharide (LPS)-treated RAW264.7 cells. In addition, this fraction suppressed proliferation and interferon (IFN)-[Formula: see text] production in splenic T lymphocytes. Cm-WE blocked the activation of nuclear factor (NF)-[Formula: see text]B and activator protein (AP)-1 and their upstream inflammatory signaling cascades, including Syk, MEK, and JNK. Using kinase assays, Syk was identified as the target enzyme most strongly inhibited by Cm-WE. These results strongly suggest that Cm-WE suppresses inflammatory responses by inhibiting Syk kinase activity, with potential implications for novel neutraceutic and pharmaceutic biomaterials.

  9. Genes involved in virulence of the entomopathogenic fungus Beauveria bassiana.

    Science.gov (United States)

    Valero-Jiménez, Claudio A; Wiegers, Harm; Zwaan, Bas J; Koenraadt, Constantianus J M; van Kan, Jan A L

    2016-01-01

    Pest insects cause severe damage to global crop production and pose a threat to human health by transmitting diseases. Traditionally, chemical pesticides (insecticides) have been used to control such pests and have proven to be effective only for a limited amount of time because of the rapid spread of genetic insecticide resistance. The basis of this resistance is mostly caused by (co)dominant mutations in single genes, which explains why insecticide use alone is an unsustainable solution. Therefore, robust solutions for insect pest control need to be sought in alternative methods such as biological control agents for which single-gene resistance is less likely to evolve. The entomopathogenic fungus Beauveria bassiana has shown potential as a biological control agent of insects, and insight into the mechanisms of virulence is essential to show the robustness of its use. With the recent availability of the whole genome sequence of B. bassiana, progress in understanding the genetics that constitute virulence toward insects can be made more quickly. In this review we divide the infection process into distinct steps and provide an overview of what is currently known about genes and mechanisms influencing virulence in B. bassiana. We also discuss the need for novel strategies and experimental methods to better understand the infection mechanisms deployed by entomopathogenic fungi. Such knowledge can help improve biocontrol agents, not only by selecting the most virulent genotypes, but also by selecting the genotypes that use combinations of virulence mechanisms for which resistance in the insect host is least likely to develop.

  10. Bestrijding van de slawortelboorder Hepialus lupulinus met de insectenpathogene schimmel Beauveria bassiana : praktijkproeven pioenroos 2004 - 2005

    NARCIS (Netherlands)

    Boertjes, B.C.

    2005-01-01

    Uit het onderzoek kan geconcludeerd worden dat Beauveria bassiana een goede bestrijder is van de slawortelboorder Hepialus lupulinus L. Door de schimmel aangetaste rupsen zijn makkelijk aan te treffen in en buiten het behandelde perceel. Doordat in deze proeven Beauveria bassiana ook werd aangetroff

  11. Laboratory Study on Biological Control of Ticks (Acari: Ixodidae by Entomopathogenic Indigenous Fungi (Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    M Abdigoudarzi

    2009-12-01

    Full Text Available Background: Chemical control method using different acaricides as spray, dipping solution or pour-on is routinely used for controlling ticks. Biological control agents are favorable due to their safety for animals and environment. Entomopathogenic fungi such as Beauveria bassiana are well known for controlling ticks. In this study, two Iranian indigenous strains of B. bassiana (B. bassiana 5197 and B. bassiana Evin were selected and grown on specific me­dia. The pathogenic effects of these strains were evaluated on adult stages of two Iranian Ixodidae members (H. anatolicum anatolicum Koch 1844, and H. marginatum Koch 1844 by dipping method.Methods: Two Iranian strains of Beauveria bassiana (Beauveria bassiana 5197 and Beauveria bassiana Evin were selected and were grown successfully on specific media. The pathogenic effects of these strains were evaluated on adult stages of Iranian Ixodidae members such as, Hyalomma anatolicum anatolicum and H. marginatum by dipping method (these ticks were grown up at laboratory conditions during 2002 up to 2003 and still it is continued .Results: There was no effect of strain 5197 on mortality or fecundity rates for ticks. There was acute phase sign of paralysis in test group after dipping ticks in suspension made from Evin strain of B. bassiana. In addition, the test groups were totally died after four months, but the control groups survived for six months.Conclusion: High concentration of fungal spores is needed for inducing fungal infection. Additional study using different strains and fungi on Iranian ticks is proposed. 

  12. Characterization of Clostridium perfringens isolates obtained from 2010 to 2012 from chickens with necrotic enteritis in Korea.

    Science.gov (United States)

    Park, Ji Young; Kim, Sara; Oh, Jae Young; Kim, Hye Ryoung; Jang, Il; Lee, Hee Soo; Kwon, Yong Kuk

    2015-06-01

    Clostridium perfringens produces diverse virulent toxins that cause necrotic enteritis in poultry, resulting in a great negative impact on the poultry industry. To study the characteristics of C. perfringens in chickens, we isolated 88 strains from chickens (1 strain per flock) with necrotic enteritis. The isolated bacterial strains were screened for toxin type and antimicrobial susceptibility. Necropsy of 17 chickens that died from necrotic enteritis revealed that their intestines were dilated with inflammatory exudates and characterized by mucosal necrosis. All the isolated strains were identified as toxin type A using multiplex PCR for toxin typing. We found that the rate of netB-positive strains isolated from dead chickens was significantly higher (8 of 17) than the rate among healthy chickens (2 of 50). We performed antimicrobial susceptibility test with 20 selected antimicrobial agents using the disk diffusion test and found that 30 tested strains were completely resistant to 5 antibiotics and partially resistant to 6 antibiotics whereas all the strains were susceptible to 9 antimicrobial agents. Using pulsed-field gel electrophoresis analysis, the 17 strains were divided into 13 genetic clusters showing high genetic diversity. In conclusion, C. perfringens strains isolated from Korean poultry showed a high resistance to antimicrobial drugs and high genetic diversity, suggesting that continuous monitoring is essential to prevent outbreaks of necrotic enteritis in chickens.

  13. Genetic polymorphisms and drug susceptibility in four isolates of Leishmania tropica obtained from Canadian soldiers returning from Afghanistan.

    Directory of Open Access Journals (Sweden)

    Marie Plourde

    2012-01-01

    Full Text Available BACKGROUND: Cutaneous leishmaniasis (CL is a vector-borne parasitic disease characterized by the presence of one or more lesions on the skin that usually heal spontaneously after a few months. Most cases of CL worldwide occur in Southwest Asia, Africa and South America, and a number of cases have been reported among troops deployed to Afghanistan. No vaccines are available against this disease, and its treatment relies on chemotherapy. The aim of this study was to characterize parasites isolated from Canadian soldiers at the molecular level and to determine their susceptibility profile against a panel of antileishmanials to identify appropriate therapies. METHODOLOGY/PRINCIPAL FINDINGS: Parasites were isolated from skin lesions and characterized as Leishmania tropica based on their pulsed field gel electrophoresis profiles and pteridine reductase 1 (PTR1 sequences. Unusually high allelic polymorphisms were observed at several genetic loci for the L. tropica isolates that were characterized. The drug susceptibility profile of intracellular amastigote parasites was determined using an established macrophage assay. All isolates were sensitive to miltefosine, amphotericin B, sodium stibogluconate (Pentostam and paromomycin, but were not susceptible to fluconazole. Variable levels of susceptibility were observed for the antimalarial agent atovaquone/proguanil (Malarone. Three Canadian soldiers from this study were successfully treated with miltefosine. CONCLUSIONS/SIGNIFICANCE: This study shows high heterogeneity between the two L. tropica allelic versions of a gene but despite this, L. tropica isolated from Afghanistan are susceptible to several of the antileishmanial drugs available.

  14. Antifungal susceptibility and molecular typing of 115 Candida albicans isolates obtained from vulvovaginal candidiasis patients in 3 Shanghai maternity hospitals.

    Science.gov (United States)

    Ying, Chunmei; Zhang, Hongju; Tang, Zhenhua; Chen, Huifen; Gao, Jing; Yue, Chaoyan

    2016-05-01

    In our multicenter study, we studied the distribution of Candida species in vulvovaginal candidiasis patients and investigated antifungal susceptibility profile and genotype of Candida albicans in vaginal swab. A total of 115 Candida albicans strains were detected in 135 clinical isolates. Minimum inhibitory concentration determinations showed that 83% and 81% of the 115 Candida albicans strains were susceptible to fluconazole and voriconazole. Randomly amplified polymorphic DNA analysis (RAPD) was applied to identify clonally related isolates from different patients at the local level. All tested strains were classified into genotype A (77.4%), genotype B (18.3%), and genotype C (4.3%). Genotype A was further classified into five subtypes and genotype B into two subtypes.Candida albicans was the dominant pathogen of vulvovaginal candidiasis, the majority belonging to genotype A in this study. Exposure to azoles is a risk factor for the emergence of azole resistance among Candida albicans isolated from VVC patients.

  15. Genetic Diversity of Clostridium sporogenes PA 3679 Isolates Obtained from Different Sources as Resolved by Pulsed-Field Gel Electrophoresis and High-Throughput Sequencing.

    Science.gov (United States)

    Schill, Kristin M; Wang, Yun; Butler, Robert R; Pombert, Jean-François; Reddy, N Rukma; Skinner, Guy E; Larkin, John W

    2015-10-30

    Clostridium sporogenes PA 3679 is a nonpathogenic, nontoxic model organism for proteolytic Clostridium botulinum used in the validation of conventional thermal food processes due to its ability to produce highly heat-resistant endospores. Because of its public safety importance, the uncertain taxonomic classification and genetic diversity of PA 3679 are concerns. Therefore, isolates of C. sporogenes PA 3679 were obtained from various sources and characterized using pulsed-field gel electrophoresis (PFGE) and whole-genome sequencing. The phylogenetic relatedness and genetic variability were assessed based on 16S rRNA gene sequencing and whole-genome single nucleotide polymorphism (SNP) analysis. All C. sporogenes PA 3679 isolates were categorized into two clades (clade I containing ATCC 7955 NCA3679 isolates 1961-2, 1990, and 2007 and clade II containing PA 3679 isolates NFL, UW, FDA, and Campbell and ATCC 7955 NCA3679 isolate 1961-4). The 16S maximum likelihood (ML) tree clustered both clades within proteolytic C. botulinum strains, with clade I forming a distinct cluster with other C. sporogenes non-PA 3679 strains. SNP analysis revealed that clade I isolates were more similar to the genomic reference PA 3679 (NCTC8594) genome (GenBank accession number AGAH00000000.1) than clade II isolates were. The genomic reference C. sporogenes PA 3679 (NCTC8594) genome and clade I C. sporogenes isolates were genetically distinct from those obtained from other sources (University of Wisconsin, National Food Laboratory, U.S. Food and Drug Administration, and Campbell's Soup Company). Thermal destruction studies revealed that clade I isolates were more sensitive to high temperature than clade II isolates were. Considering the widespread use of C. sporogenes PA 3679 and its genetic information in numerous studies, the accurate identification and genetic characterization of C. sporogenes PA 3679 are of critical importance. Copyright © 2015, American Society for Microbiology. All

  16. Drug Susceptibility and Genetic Evaluation of Plasmodium falciparum Isolates Obtained in Four Distinct Geographical Regions of Kenya

    OpenAIRE

    Mbaisi, Abigael; Liyala, Pamela; Eyase, Fredrick; Achilla, Rachel; Akala, Hosea; Wangui, Julia; Mwangi, Josphat; Osuna, Finnley; Alam, Uzma; Smoak, Bonnie L.; Davis, Jon M.; Kyle, Dennis E.; Coldren, Rodney L; Mason, Carl; Waters, Norman C.

    2004-01-01

    The drug resistance profiles of Plasmodium falciparum isolated from four regions in Kenya were analyzed for drug resistance profiles. We observed variability in resistance to a broad range of antimalarial drugs across Kenya as determined from in vitro drug susceptibility screening and genotyping analysis.

  17. Up-regulation of carbon metabolism-related glyoxylate cycle and toxin production in Beauveria bassiana JEF-007 during infection of bean bug, Riptortus pedestris (Hemiptera: Alydidae).

    Science.gov (United States)

    Yang, Yi-Ting; Lee, Se Jin; Nai, Yu-Shin; Kim, Sihyeon; Kim, Jae Su

    2016-10-01

    Beauveria bassiana (Bb) is used as an environment-friendly biopesticide. However, the molecular mechanisms of Bb-host interactions are not well understood. Herein, RNA isolated from B. bassiana (Bb JEF-007) and Riptortus pedestris (Hemiptera: Alydidae) infected with this strain were firstly subjected to high-throughput next generation sequencing (NGS) to analyze and compare transcriptomes. Due to lack of fungal and host genome information, fungal transcriptome was processed to partially exclude non-infection specific genes and host-flora. Differentially Expressed Gene (DEG) analysis showed that 2381 genes were up-regulated and 2303 genes were down-regulated upon infection. Most DEGs were classified into the categories of single-organism, cellular and metabolism processes by Gene Ontology analysis. Most DEGs were involved in metabolic pathways based on Kyoto Encyclopedia of Genes and Genomes pathway mapping. Carbon metabolism-related enzymes in the glyoxylate cycle were significantly up-regulated, suggesting a possible role for them in Bb growth in the host. Additionally, transcript levels of several fungal genes were dramatically increased after infection, such as cytotoxic lectin-like protein, bacterial-like toxin, proteins related to cell wall formation, hyphal growth, nutrient uptake, and halogenated compound synthesis. This work provides insight into how entomopathogenic B. bassiana grows in agriculturally harmful bean bug at 6 d post infection.

  18. Isolation of entomopathogenic fungi from Northern Thailand and their production in cereal grains.

    Science.gov (United States)

    Mar, Thet Thet; Suwannarach, Nakarin; Lumyong, Saisamorn

    2012-12-01

    Spore productivity in six entomopathogenic fungal strains isolated from insect cadavers at four locations in Chiang Mai province was evaluated in five cereal grains: white-rice, wheat, rye, corn and sorghum. According to sequence analysis of the internal transcribed spacer regions of these isolates, they were closely related to Beauveria bassiana (2 isolates), Metarhizium flavoviride (1 isolate), Metarhizium anisopliae (1 isolate), Paecilomyces lilacinus (1 isolate) and Isaria tenuipes (1 isolate). Among all fungal isolates, the maximum amount of spores (530.0 × 10(9) conidia/g) was yielded P. lilacinus CMUCDMT02 on sorghum grain followed by white-rice (399.3 × 10(9) conidia/g). Moreover, the highest number of spore in M. flavoviride was 102.8 × 10(9) conidia/g sorghum whereas white-rice yielded the greatest amount of spore for B. bassiana CMUCDMF03 (141.0 × 10(9) conidia/g) after 60 days incubation. The fungal growth rate was found highest in corn for all strains and rye showed the lowest with the exception of P. lilacinus CMUCDMT02 among the tested grains. Spore viability was over 80 % for all isolates that had been inoculated for 60 days. Fungal conidia suspension of P. lilacinus obtained highest virulence against Bactrocera spp. at a concentration of 1 × 10(6) spore/ml. The strains isolated, exhibited good production of conidia suggesting a promising strategy for the mass production of inoculum as biocontrol agents with low production cost.

  19. Frequency of MCR-1-mediated colistin resistance among Escherichia coli clinical isolates obtained from patients in Canadian hospitals (CANWARD 2008-2015).

    Science.gov (United States)

    Walkty, Andrew; Karlowsky, James A; Adam, Heather J; Lagacé-Wiens, Philippe; Baxter, Melanie; Mulvey, Michael R; McCracken, Melissa; Poutanen, Susan M; Roscoe, Diane; Zhanel, George G

    2016-01-01

    Colistin is often used as an antimicrobial of last resort for the treatment of infections caused by multidrug-resistant gram-negative bacilli. In 2015, plasmid-mediated colistin resistance in Escherichia coli due to MCR-1 was described. The purpose of this study was to evaluate the frequency of colistin resistance among E. coli clinical isolates obtained from patients in Canadian hospitals as part of the Canadian Ward Surveillance Study (CANWARD) and to determine how often the mcr-1 gene is detected among the colistin-resistant subset. From January 2008 to December 2015 (excluding 2011), 10 to 15 sentinel hospitals submitted consecutive clinical isolates (1 per patient per infection site) from blood (100-240), respiratory (100-150), urine (25-100) and wound (25-100) infections. We performed susceptibility testing using Clinical and Laboratory Standards Institute broth microdilution methods. Isolates that showed resistance to colistin as defined by European Committee on Antimicrobial Susceptibility Testing breakpoints (minimum inhibitory concentration ≥ 4 µg/mL) were evaluated for the mcr-1 gene by polymerase chain reaction. In total, 5571 E. coli clinical isolates were obtained over the study years. Twelve isolates (0.2%) were resistant to colistin. The proportion of colistin-resistant isolates varied from 0.0% to 0.5% depending on the study year, and there was no clear trend toward increasing resistance over time. Typically the colistin-resistant isolates remained susceptible to antimicrobials from several other classes. Two colistin-resistant isolates (0.04%) were found to harbour the mcr-1 gene. The results suggest that colistin resistance among E. coli human clinical isolates, including resistance mediated by the mcr-1 gene, remains rare in Canada.

  20. Phylogenetic and biogeographic implications inferred by mitochondrial intergenic region analyses and ITS1-5.8S-ITS2 of the entomopathogenic fungi Beauveria bassiana and B. brongniartii

    Directory of Open Access Journals (Sweden)

    Typas Milton A

    2010-06-01

    Full Text Available Abstract Background The entomopathogenic fungi of the genus Beauveria are cosmopolitan with a variety of different insect hosts. The two most important species, B. bassiana and B. brongniartii, have already been used as biological control agents of pests in agriculture and as models for the study of insect host - pathogen interactions. Mitochondrial (mt genomes, due to their properties to evolve faster than the nuclear DNA, to contain introns and mobile elements and to exhibit extended polymorphisms, are ideal tools to examine genetic diversity within fungal populations and genetically identify a species or a particular isolate. Moreover, mt intergenic region can provide valuable phylogenetic information to study the biogeography of the fungus. Results The complete mt genomes of B. bassiana (32,263 bp and B. brongniartii (33,920 bp were fully analysed. Apart from a typical gene content and organization, the Beauveria mt genomes contained several introns and had longer intergenic regions when compared with their close relatives. The phylogenetic diversity of a population of 84 Beauveria strains -mainly B. bassiana (n = 76 - isolated from temperate, sub-tropical and tropical habitats was examined by analyzing the nucleotide sequences of two mt intergenic regions (atp6-rns and nad3-atp9 and the nuclear ITS1-5.8S-ITS2 domain. Mt sequences allowed better differentiation of strains than the ITS region. Based on mt and the concatenated dataset of all genes, the B. bassiana strains were placed into two main clades: (a the B. bassiana s. l. and (b the "pseudobassiana". The combination of molecular phylogeny with criteria of geographic and climatic origin showed for the first time in entomopathogenic fungi, that the B. bassiana s. l. can be subdivided into seven clusters with common climate characteristics. Conclusions This study indicates that mt genomes and in particular intergenic regions provide molecular phylogeny tools that combined with criteria of

  1. Antimicrobial Resistance Pattern in Escherichia coli Isolates Obtained from a Specialized Women and Children Hospital in Shiraz, Iran: A Prevalence Study

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    Mahtab Hadadi

    2016-10-01

    Full Text Available Abstract Background: Escherichia coli, known as a clinically significant bacteria, can cause a wide range of infections, including urinary tract infections (UTIs, blood stream infections (BSIs, and can frequently be isolated from various clinical specimens. Evaluation of antimicrobial resistant pattern is a necessary action, especially about such bacteria which are frequent and life threatening. The aim of this study was to determine the frequency and antimicrobial resistance pattern of E. coli isolates obtained from various clinical specimens. Methods: This retrospective study was performed within a seven month period from January 2015 to August 2015 at a specialized women and children hospital in Shiraz, Iran. E. coli isolates were obtained from various clinical specimens and identified using standard microbiological procedure. Antimicrobial susceptibility patterns were determined using disk diffusion method in accordance with CLSI recommendation. Results: Of the total 130 positive cultures, the majority of E. coli isolates were obtained from urine (96=73.8% and blood (11=8.5% specimens. Overall, gentamicin (70.8% was the effective antibiotic for the tested E. coli isolates. E. coli isolates obtained from urine specimens showed the highest resistance rates against ampicillin (84.4% and nalidixic acid (61.5%; while they showed the most sensitivity to gentamicin (79.2%, nitrofurantoin (70.8% and ciprofloxacin (66.7%. Moreover, the highest antibiotic resistance rates belonged to the isolates recovered from endotracheal tube (ETT. Conclusion: The results showed that gentamicin was the most effective antibiotic against E. coli infections. However, in addition to the gentamicin, we can recommend nitrofurantoin and ciprofloxacin as the other effective agents for UTIs

  2. SCREENING FOR ANTIBACTERIAL PROPERTIES OF THREE MEDICINAL PLANTS AGAINST SALMONELLA SP. ISOLATES OBTAINED FROM BROILER CARCASS IN INDONESIA

    Directory of Open Access Journals (Sweden)

    M. Poeloengan

    2013-03-01

    Full Text Available Antibacterial properties of three medicinal plants used widely amongst the native Indonesians, i.e., sirih (Piper betel, garlic (Allium sativum and jinten hitam (Nigella sativa were screened and evaluated against Salmonella sp. isolates. For this purpose, the three plants’ powder, aqueous and ethanol extracts were prepared. The tests’ results demonstrated the three plants’ anti Salmonella sp. activities. In this conjunction, the aqueous extract of Allium sativum, the ethanol extracts of Nigella sativa, and Piper betel, consecutively produced 17 mm, 15 mm and 13 mm diameter of bacterial growth inhibition zones. As a comparison, sensitivity tests of three commercial antibiotics, i.e., chloramphenicol, tetracycline, and gentamycin on Salmonella sp. isolates produced 19.7-27.3 mm growth inhibition zones. Garlic having antimicrobial potential was comparable to the commercial antibiotics, can be used as a decontaminant against Salmonella sp. to maintain the quality of the broiler carcasses and therefore prolonging the carcass shelf-life.

  3. Risk factors for isolation of Staphylococcus aureus or Streptococcus dysgalactiae from milk culture obtained approximately 6 days post calving.

    Science.gov (United States)

    Osterås, Olav; Whist, Anne Cathrine; Sølverød, Liv

    2008-02-01

    Milk culture results at approximately 6 d post calving were assessed in a 2-year retrospective single-cohort study in 178 Norwegian herds. A combined teat dipping and selective antibiotic therapy trial was performed in these herds where cows with composite milk somatic cell count (CMSCC) >100,000 cells/ml before drying-off (geometric mean of the last three CMSCC test-days) and isolation of Staphylococcus aureus or Streptococcus dysgalactiae were selected for either short-acting lactation antibiotic treatment or long-acting dry cow antibiotic treatment. Milk culture results at approximately 6 d post-calving were available from 437 treated cows and 3061 non-treated cows before drying-off and separate multivariable logistic regression models were ran for these two groups. Risk factors associated with isolation of Staph. aureus 6 d post calving for non-treated cows were CMSCC >400,000 cells/ml before drying-off v. 200,000 cells/ml before drying-off v. 50,000 cells/ml compared with teat disinfection (PMTD) did not influence the isolation of Staph. aureus 6 d post calvin, but it was less likely to isolate Str. dysgalactiae 6 d post calving if iodine PMTD was used regularly rather than irregularly. The external teat sealant had no effect on either of the two bacteria. This study indicates that the CMSCC limit for sampling cows before drying-off can be reduced to 50,000 cells/ml in herds with a Str. dysgalactiae problem. Iodine PMTD should also be recommended in these herds. Cows with a CMSCC > 400,000 cells/ml prior to drying-off should receive long-acting dry cow formula irrespective of the milk result.

  4. Characterization of Nivalenol-Producing Fusarium culmorum Isolates Obtained from the Air at a Rice Paddy Field in Korea

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    Da-Woon Kim

    2016-06-01

    Full Text Available Together with the Fusarium graminearum species complex, F. culmorum is a major member of the causal agents of Fusarium head blight on cereals such as wheat, barley and corn. It causes significant yield and quality losses and results in the contamination of grain with mycotoxins that are harmful to humans and animals. In Korea, F. culmorum is listed as a quarantine fungal species since it has yet to be found in the country. In this paper, we report that two isolates (J1 and J2 of F. culmorum were collected from the air at a rice paddy field in Korea. Species identification was confirmed by phylogenetic analysis using multi-locus sequence data derived from five genes encoding translation elongation factor, histone H3, phosphate permease, a reductase, and an ammonia ligase and by morphological comparison with reference strains. Both diagnostic PCR and chemical analysis confirmed that these F. culmorum isolates had the capacity to produce nivalenol, the trichothecene mycotoxin, in rice substrate. In addition, both isolates were pathogenic on wheat heads and corn stalks. This is the first report on the occurrence of F. culmorum in Korea.

  5. Baeuveria bassiana (BalsVuill and Metarhizium anisioplae (Metsch. Sorokin in the pupas control of Prodiplosis longifila Gagné on asparagus crop

    Directory of Open Access Journals (Sweden)

    Carolina Cedano

    2012-03-01

    Full Text Available The objective of the present work was to evaluate the effect of the entomopathogen fungi Baeuveria bassiana and Metarhizium anisioplae of the Prodiplosis longifila pupas, in order to reduce the adult population of this insect. The treatments consisted on the application of two different propagel concentrations (mycel and conidia of each entomopathogen. One of these concentration was 1x106 propagels/ml (obtained from the total amount of conidia collected of 25 kg of rice colonized by the entomopathogen; and the other one, was 1x107 propagels/ml (obtained from the total amount of conidia collected of 40 kg of rice colonized by the entomopathogen. As a check a non application treatment was considered. The applications were trough the irrigation system and started 15 days after the end of harvest and were repeated each five days during a month, coincident with the period of most falls of pupas of the soil. As a result Baeuveria bassiana at 1x107 propagels/ml shown 53.4 % of the total pupas colonized by the entomopathogen, which allows indicating B. bassiana as a promissory biocontrol of this specie

  6. Comparative evaluation of five Beauveria isolates for housefly (Musca domestica L.) control and growth optimization of selected strain.

    Science.gov (United States)

    Mishra, Sapna; Malik, Anushree

    2012-11-01

    Pathogenic potential of five native Beauveria isolates was assessed against housefly adult and larvae in laboratory bioassays. Beauveria isolate Beauveria bassiana HQ917687 showed highest virulence with 72.3 and 100 % mortality of larvae and adults of Musca domestica, respectively. Other Beauveria isolates caused 36-52 % housefly larval mortality while the adult mortalities varied between 72 and 82 %. B. bassiana HQ917687 also showed the fastest killing activity with LT(50) of 4 days (for larvae) and 3 days (for adults). This isolate showing highest virulence was selected for its growth optimization in terms of biomass and spore production using response surface methodology. The optimum value of temperature, yeast extract, and pH for maximum biomass and spore production was predicted as 27 °C, 5.00 g/l, and 6.75, respectively. Temperature was found to be the most critical factor influencing biomass and spore yield of the fungus and even nullified the effects of other factors at sufficiently higher value. The results obtained in this study depict the significance of appropriate strain selection and process parameter optimization in order to facilitate mass production of biocontrol agents.

  7. Evaluation of a New Entomopathogenic Strain of Beauveria bassiana and a New Field Delivery Method against Solenopsis invicta

    Science.gov (United States)

    Li, Jun; Guo, Qiang; Lin, Miaofeng; Jiang, Lu; Ye, Jingwen; Chen, Dasong; Li, Zhigang; Dai, Jianqing; Han, Shichou

    2016-01-01

    Solenopsis invicta Buren is one of the most important pests in China, and control measures are mainly based on the use of synthetic pesticides, which may be inadequate and unsustainable. Hence, there is a growing interest in developing biological control alternatives for managing S. invicta, such as the use of entomopathogenic fungi. To facilitate the commercialization of entomopathogenic fungi against S. invicta, 10 Beauveria bassiana isolates originating from different hosts were tested for virulence in laboratory bioassays, and the most pathogenic strain, ZGNKY-5, was tested in field studies using an improved pathogen delivery system. The cumulative mortality rate reached 93.40% at 1×108 mL-1 conidia after 504 h. The germination and invasion of the spores were observed under a scanning electron microscope, and several conidia adhered to the cuticle of S. invicta after 2 h. Furthermore, the germ tubes of the conidia oriented toward the cuticle after 48 h, and the mycelium colonized the entire body after 96 h. Based on the efficacy observed in the laboratory trials, further experiments were performed with ZGNKY-5 strain to evaluate its utility in an injection control technology against S. invicta in the field. We found that three dosage treatments of ZGNKY-5 strain (500 mL, 750 mL, and 1,000 mL per nest) had significant control effects. Our results show that this strain of Beauveria bassiana and our control method were effective against S. invicta in both laboratory and field settings. PMID:27341441

  8. Evaluation of a New Entomopathogenic Strain of Beauveria bassiana and a New Field Delivery Method against Solenopsis invicta.

    Science.gov (United States)

    Li, Jun; Guo, Qiang; Lin, Miaofeng; Jiang, Lu; Ye, Jingwen; Chen, Dasong; Li, Zhigang; Dai, Jianqing; Han, Shichou

    2016-01-01

    Solenopsis invicta Buren is one of the most important pests in China, and control measures are mainly based on the use of synthetic pesticides, which may be inadequate and unsustainable. Hence, there is a growing interest in developing biological control alternatives for managing S. invicta, such as the use of entomopathogenic fungi. To facilitate the commercialization of entomopathogenic fungi against S. invicta, 10 Beauveria bassiana isolates originating from different hosts were tested for virulence in laboratory bioassays, and the most pathogenic strain, ZGNKY-5, was tested in field studies using an improved pathogen delivery system. The cumulative mortality rate reached 93.40% at 1×108 mL-1 conidia after 504 h. The germination and invasion of the spores were observed under a scanning electron microscope, and several conidia adhered to the cuticle of S. invicta after 2 h. Furthermore, the germ tubes of the conidia oriented toward the cuticle after 48 h, and the mycelium colonized the entire body after 96 h. Based on the efficacy observed in the laboratory trials, further experiments were performed with ZGNKY-5 strain to evaluate its utility in an injection control technology against S. invicta in the field. We found that three dosage treatments of ZGNKY-5 strain (500 mL, 750 mL, and 1,000 mL per nest) had significant control effects. Our results show that this strain of Beauveria bassiana and our control method were effective against S. invicta in both laboratory and field settings.

  9. Evaluation of a New Entomopathogenic Strain of Beauveria bassiana and a New Field Delivery Method against Solenopsis invicta.

    Directory of Open Access Journals (Sweden)

    Jun Li

    Full Text Available Solenopsis invicta Buren is one of the most important pests in China, and control measures are mainly based on the use of synthetic pesticides, which may be inadequate and unsustainable. Hence, there is a growing interest in developing biological control alternatives for managing S. invicta, such as the use of entomopathogenic fungi. To facilitate the commercialization of entomopathogenic fungi against S. invicta, 10 Beauveria bassiana isolates originating from different hosts were tested for virulence in laboratory bioassays, and the most pathogenic strain, ZGNKY-5, was tested in field studies using an improved pathogen delivery system. The cumulative mortality rate reached 93.40% at 1×108 mL-1 conidia after 504 h. The germination and invasion of the spores were observed under a scanning electron microscope, and several conidia adhered to the cuticle of S. invicta after 2 h. Furthermore, the germ tubes of the conidia oriented toward the cuticle after 48 h, and the mycelium colonized the entire body after 96 h. Based on the efficacy observed in the laboratory trials, further experiments were performed with ZGNKY-5 strain to evaluate its utility in an injection control technology against S. invicta in the field. We found that three dosage treatments of ZGNKY-5 strain (500 mL, 750 mL, and 1,000 mL per nest had significant control effects. Our results show that this strain of Beauveria bassiana and our control method were effective against S. invicta in both laboratory and field settings.

  10. Detection of sul1, sul2 and sul3 in sulphonamide resistant Escherichia coli isolates obtained from healthy humans, pork and pigs in Denmark

    DEFF Research Database (Denmark)

    Hammerum, Anette Marie; Sandvag, Dorthe; Andersen, Sigrid R.

    2006-01-01

    The occurrence of sulphonamide resistance was investigated in 998 Escherichia coli isolates, obtained from pig faeces collected at slaughter, Danish pork collected at retail outlets and from faeces from healthy persons in Denmark. In total 18% (n = 35), 20% (n = 38) and 26% (n = 161) of the E. co...

  11. Nutritional evaluation of the germ meal and its protein isolate obtained from the carob seed (Ceratonia siliqua) in the rat.

    Science.gov (United States)

    Drouliscos, N J; Malefaki, V

    1980-01-01

    1. Evaluation of the germ meal (CGM) of carob seed (Ceratonia siliqua) and its protein isolate was carried out with weanling rats. Comparisons were made with casein, soya-bean meal, whole defatted egg and a soya-bean protein isolate (Promine-D) as protein sources. The growth-promoting effects and certain biological indices were evaluated using the protein efficiency ratio (PER), biological value (BV) and net protein utilization (NPU) bioassay procedures. 2. The unsupplemented CGM had a PER of 1.66 +/- 0.09 and an NPU of 0.58 +/- 0.013. Addition of DL-methionine at 4, 8 and 12 g/kg diet resulted in a PER of 1.95 +/- 0.11, 2.01 +/- 0.11 and 1.90 +/- 0.11 respectively. The corresponding BV values were 0.80 +/- 0.003, 0.78 +/- 0.015 and 0.74 +/- 0.011, and those for NPU 0.69 +/- 0.013, 0.66 +/- 0.026 and 0.63 +/- 0.020 respectively. The addition of amino acids improved the PER (2.24--2.59), BV (0.78--0.79) and NPU (0.71--0.73) values. 3. The BV and NPU assays for the unsupplemented carob germ isolate were low (BV 0.36 +/- 0.016, NPU 0.35 +/- 0.015). Supplementation with amino acids resulted in a positive increase with values of 0.66 +/- 0.013 and 0.64 +/- 0.013 for BV and NPU respectively.

  12. Identification of bacterial isolates obtained from intestinal contents associated with 12,000-year-old mastodon remains.

    Science.gov (United States)

    Rhodes, A N; Urbance, J W; Youga, H; Corlew-Newman, H; Reddy, C A; Klug, M J; Tiedje, J M; Fisher, D C

    1998-02-01

    Mastodon (Mammut americanum) remains unearthed during excavation of ancient sediments usually consist only of skeletal material, due to postmortem decomposition of soft tissues by microorganisms. Two recent excavations of skeletal remains in anoxic sediments in Ohio and Michigan, however, have uncovered organic masses which appear to be remnants of the small and large intestines, respectively. Macrobotanical examinations of the composition of these masses revealed assemblages of plant material radiocarbon dated to approximately 11,500 years before the present and thought to be incompletely digested food remains from this extinct mammal. We attempted to cultivate and identify bacteria from the intestinal contents, bone-associated sediments, and sediments not in proximity to the remains using a variety of general and selective media. In all, 295 isolates were cultivated, and 38 individual taxa were identified by fatty acid-methyl ester (FAME) profiles and biochemical characteristics (API-20E). The taxonomic positions of selected enteric and obligately anaerobic bacteria were confirmed by 16S ribosomal DNA (rDNA) sequencing. Results indicate that the intestinal and bone-associated samples contained the greatest diversity of bacterial taxa and that members of the family Enterobacteriaceae represented 41% of all isolates and were predominant in the intestinal masses and sediments in proximity to the skeleton but were uncommon in the background sediments. Enterobacter cloacae was the most commonly identified isolate, and partial rDNA sequencing revealed that Rahnella aquatilis was the correct identity of strains suggested by FAME profiles to be Yersinia enterocolitica. No Bacteroides spp. or expected intestinal anaerobes were recovered. The only obligate anaerobes recovered were clostridia, and these were not recovered from the small intestinal masses. Microbiological evidence from this study supports other, macrobotanical data indicating the intestinal origin of these

  13. Screening Test of Highly Virulent Strains of Entomopathogenic Fungi Beauveria bassiana against Apriona germari Larvae%桑天牛幼虫高毒力白僵菌菌株筛选

    Institute of Scientific and Technical Information of China (English)

    李会平; 黄大庄; 王志刚; 闫海霞; 郑建伟

    2007-01-01

    为利用白僵菌防治桑天牛,从野外感病死亡昆虫体上进行分离,同时利用黄粉虫对河北省土壤中的白僵菌进行诱集.诱集结果表明:白僵菌较多存在于自然的或人为活动较少的土壤中.利用获得的菌株对桑天牛幼虫进行生物测定,初步筛选出Bi05和Bs04 2个致病力较强的菌株,其接种10 d后的死亡率分别为96.47% 和92.93%,致死中时分别为4.53 d和4.83 d,而其他菌株的死亡率在28.59%和 82.12%之间,致死中时从5.84 d到16.4 d.利用不同孢子浓度的悬浮液对桑天牛幼虫进行接种,进一步测定了Bi05和Bs04菌株的致死中浓度分别为6.76×105 和1.39×106 conidia·mL-1.Bi05和Bs04菌株具有生物防治桑天牛的潜力.%To develop microbial control of Apriona germari, a destructive stem-boring pest, Beauveria bassiana were isolated from infected insects obtained in the field and by the bait method from soil samples in Hebei Province. The bait results showed that B. bassiana occurred more frequently in the natural and undestroyed soils.The virulence of all isolates was tested in bioassays by dipping larvae in 1×108 conidia·mL-1 suspensions. Two isolates Bi05 and Bs04 with the shortest LT50 value(4.53 d and 4.83 d) caused 96.47% and 92.93% mortality(due to B. bassiana)respectively 10 d post-inoculation while other isolates only caused mortalities from 28.59% to 82.12% with LT50 value ranged from 5.84 d to 16.4 d. The virulence of Bi05 and Bs04 was further tested by exposing A. germari larvae to conidial serial concentrations of 1×104~1×108 conidia·mL-1.LC50 value of Bi05 and Bs04 were 6.76×105 and 1.39×106 conidia·mL-1, respectively. The present results suggest that the isolate Bi05 and Bs04 has excellent potential for biological control of A. germari.

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Isolation and identification of zoonotic species of genus arcobacter from chicken viscera obtained from retail distributors of the metropolitan area of San José, Costa Rica.

    Science.gov (United States)

    Villalobos, Edgar García; Jaramillo, Heriberto Fernández; Ulate, Carolina Chaves; Echandi, María Laura Arias

    2013-05-01

    Arcobacter is a genus of growing importance worldwide; some of its species are considered emerging enteropathogens and potential zoonotic agents. In Costa Rica, as well as in other countries, its isolation has been reported, so the objective of this project was to evaluate and identify the presence of Arcobacter in chicken viscera sold in the metropolitan area of San José, Costa Rica, as well as to determine the antimicrobial resistance patterns associated with it. One hundred fifty samples of chicken viscera including heart, liver, and other gastrointestinal organs were purchased from 15 supermarkets and 15 local retailers. De Boer and Houf broths were used as enrichment media; isolation was done with Arcobacter-selective medium and with membrane filtration with blood agar. Typical colonies were identified with genus-specific PCR, and species identification was made with multiplex PCR. Susceptibility to ampicillin, ciprofloxacin, chloramphenicol, erythromycin, gentamicin, and tetracycline was done with the Epsilometer test. The isolation frequency of Arcobacter genus obtained in this study was of 17.3%. A total of 33 isolates were obtained from the poultry samples, and according to the multiplex PCR methodology, 22 (66.7%) isolates were identified as Arcobacter butzleri, 8 (24.2%) as Arcobacter cryaerophilus, and 1 (3.1%) as Arcobacter skirrowii. Two strains were not identified. No statistical significant difference was found when the source of samples was compared. Resistance toward chloramphenicol was 68.75%, followed by ampicillin (43.75%) and ciprofloxacin (18.75%); all strains were susceptible to tetracycline.

  16. Laboratory Study on Biological Control of Ticks (Acari: Ixodidae by Entomopathogenic Indigenous Fungi (Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    M Abdigoudarzi

    2009-12-01

    Full Text Available Background: Chemical control method using different acaricides as spray, dipping solution or pour-on is routinely used for controlling ticks. Biological control agents are favorable due to their safety for animals and environment. Entomopathogenic fungi such as Beauveria bassiana are well known for controlling ticks. In this study, two Iranian indigenous strains of B. bassiana (B. bassiana 5197 and B. bassiana Evin were selected and grown on specific me­dia. The pathogenic effects of these strains were evaluated on adult stages of two Iranian Ixodidae members (H. anatolicum anatolicum Koch 1844, and H. marginatum Koch 1844 by dipping method."nMethods: Two Iranian strains of Beauveria bassiana (Beauveria bassiana 5197 and Beauveria bassiana Evin were selected and were grown successfully on specific media. The pathogenic effects of these strains were evaluated on adult stages of Iranian Ixodidae members such as, Hyalomma anatolicum anatolicum and H. marginatum by dipping method (these ticks were grown up at laboratory conditions during 2002 up to 2003 and still it is continued ."nResults: There was no effect of strain 5197 on mortality or fecundity rates for ticks. There was acute phase sign of paralysis in test group after dipping ticks in suspension made from Evin strain of B. bassiana. In addition, the test groups were totally died after four months, but the control groups survived for six months."nConclusion: High concentration of fungal spores is needed for inducing fungal infection. Additional study using different strains and fungi on Iranian ticks is proposed.   Keywords: Biological control, fungi, Beauveria bassiana, ticks, Ixodidae, Iran

  17. Morphological and Molecular Identification of aBeauveria bassiana Strain%一株球孢白僵菌的形态及分子生物学鉴定

    Institute of Scientific and Technical Information of China (English)

    刘达综; 高慧珊; 刘娜; 何洁明; 徐树兰; 郑常格; 吕思行

    2015-01-01

    利用PDA培养基,对收集自广东茂名的玉米螟(Pyrausta nubilalis)病原真菌(编号:Bba1菌株)进行分离培养。对 Bba1菌株的菌落形态、产孢结构和分生孢子形态、大小进行观察和测量,从形态学初步鉴定该菌株为球孢白僵菌(Beauveria bassiana);同时对Bba1菌株的26,s rDNA和rDNA ITS区域进行序列分析,与NCBI数据库中球孢白僵菌的同源性达到99%,以上。最终判定来自玉米螟的病原真菌Bba1菌株为球孢白僵菌(Beauveria bassiana)。%A strain(No.Bba1)was isolated using PDA culture medium from the muscardine cadaver ofPyrausta nubilalis collected from Maoming in Guangdong Province.In this study,the colony morphology,spore structure,spore shape and size of Bba1 strain were observed and measured,and it was initially identified asBeauveria bassiana.At the same time,its 26,s rDNA and rDNA ITS were sequenced and then compared with those ofBeauveria bassiana in NCBI database.The re-sults showed the homology was above 99%,.Eventually,Bba1 strain was identified asBeauveria bassiana.

  18. Ação do fungo Beauveria bassiana (Balsamo Vuillemin, 1912 sobre a fase parasitária do carrapato Anocentor nitens (Neumann, 1897 Schulze, 1937 (Acari: ixodidae Action of the fungus Beauveria bassiana (Balsamo Vuillemin, 1912 on the parasitic phase of the tick Anocentor nitens (Neumann, 1897 Schulze, 1937 (Acari: ixodidae

    Directory of Open Access Journals (Sweden)

    Silvia Gonzalez Monteiro

    2003-06-01

    Full Text Available A patogenicidade do fungo Beauveria bassiana (isolado 986 foi avaliada no desenvolvimento da fase parasitária do Anocentor nitens, acompanhando o desenvolvimento de larvas não alimentadas até o estágio adulto. Foram utilizados dois tratamentos e um grupo controle com cinco repetições cada. No tratamento 1 (T1, as orelhas dos bovinos foram banhadas com a suspensão fúngica de 10(8 conídios ml-1; no tratamento 2 (T2, as orelhas foram banhadas com água destilada e espalhante adesivo. No grupo controle (C, as orelhas foram banhadas com água destilada. Após 24 horas dos tratamentos, as orelhas de cada animal foram infestadas com 100mg de larvas. Os parâmetros analisados, em cada tratamento, foram: número e peso de fêmeas recuperadas; períodos de pré-postura, postura, incubação, eclosão e longevidade; percentual de eclosão; peso da massa de ovos e da quenógina; índices de eficiência reprodutiva e nutricional. Verificou-se que, no T1 e T2, o número de fêmeas recuperadas, após 35 dias, foi significantemente menor (PThis study evaluated the pathogenicity of the fungus Beauveria bassiana (Isolate 986 on the development of the parasitic phase of Anocentor nitens by accompanying the development of unfed larvae until the adult stage. Two treated and one control group were used with five repetitions each. In the first treated group (T1, cattle ears were bathed with a fungal suspension of 10(8 conidia ml-1, second treated group (T2, the ears were bathed with distilled water and Tween 80 1% and the control group (C, the ears were bathed with distilled water. One day after treatment, the ears of each animal were infested with 100 mg of larvae. The parameters analyzed in each treatment were: number and weight of females recovered; periods of pre-oviposition, oviposition, incubation, eclosion and longevity; percentage of eggs hatched; weight of the egg mass and of females after oviposition. We found that in the two treated groups the

  19. Molecular Typing and Virulence Gene Profiles of Enterotoxin Gene Cluster (egc)-Positive Staphylococcus aureus Isolates Obtained from Various Food and Clinical Specimens.

    Science.gov (United States)

    Song, Minghui; Shi, Chunlei; Xu, Xuebing; Shi, Xianming

    2016-11-01

    The enterotoxin gene cluster (egc) has been proposed to contribute to the Staphylococcus aureus colonization, which highlights the need to evaluate genetic diversity and virulence gene profiles of the egc-positive population. Here, a total of 43 egc-positive isolates (16.2%) were identified from 266 S. aureus isolates that were obtained from various food and clinical specimens in Shanghai. Seven different egc profiles were found based on the polymerase chain reaction (PCR) result for egc genes. Then, these 43 egc-positive isolates were further typed by multilocus sequence typing, pulsed-field gel electrophoresis (PFGE), multiple-locus variable-number tandem-repeat analysis (MLVA), and accessory gene regulatory (agr) typing. It showed that the 43 egc-positive isolates displayed 17 sequence types, 28 PFGE patterns, 29 MLVA types, and 4 agr types, respectively. Among them, the dominant clonal lineage was CC5-agr II (48.84%). Thirty toxin and 20 adhesion-associated genes were detected by PCR in egc-positive isolates. Notably, invasive toxin genes showed a high prevalence, such as 76.7% for Panton-Valentine leukocidin encoding genes, 27.9% for sec, and 23.3% for tsst-1. Most of the examined adhesion-associated genes were found to be conserved (76.7-100%), whereas the fnbB gene was only found in 8 (18.6%) isolates. In addition, 33 toxin gene profiles and 13 adhesion gene profiles were identified, respectively. Our results imply that isolates belonging to the same clonal lineage harbored similar adhesion gene profiles but diverse toxin gene profiles. Overall, the high prevalence of invasive virulence genes increases the potential risk of egc-positive isolates in S. aureus infection.

  20. Sensitivity of the Entomogenous Fungus Beauveria bassiana to Selected Plant Growth Regulators and Spray Additives

    Science.gov (United States)

    Storey, Greggory K.; Gardner, Wayne A.

    1986-01-01

    Mefluidide was the only one of four plant growth regulators that caused little to no significant inhibition of in vitro germination and growth of the entomogenous fungus Beauveria bassiana. Silaid, paclobutrazol, and flurprimidol significantly inhibited germination and growth. Mortality of fall armyworm, Spodoptera frugiperda, resulting from B. bassiana was significantly reduced when larvae were exposed to conidia plus soil treated with paclobutrazol. Larval mortality resulting from conidia plus soil treated with mefluidide did not differ significantly from mortality resulting from untreated conidia. Triton CS-7 was the only one of eight spray adjuvants that significantly inhibited germination of B. bassiana conidia. PMID:16347095

  1. Safety of Beauveria bassiana (Bals. Vuill. to Gallus domesticus L.

    Directory of Open Access Journals (Sweden)

    Jucelaine Haas-Costa

    2010-04-01

    Full Text Available Beauveria bassiana, an important insect pathogen was used to evaluate its effect on the feeding, behavior, histology of the digestory system and anatomy of Gallus domesticus. The fungus (strain Unioeste 4 was administrated orally treated with chicken food. Chicken feces were collected, and the birds observed for 28 days. Also, were evaluated the weight of the birds, as well as any sign of intoxication or pathological modification. Tissue samples were withdrawn to test lesions with the optic microscope. There was 100% of survival of the birds, and no behavior alteration or external lesion was found. The test group presented the highest weight gain (1383.9±54.4g. Viable conidia were observed in the feces until 24 h after feeding the fungus, indicating that there was no germination inside the digestory system. No tissue lesion was observed, providing overwhelming evidence for the safety of B. bassiana to non-target organism G. domesticus.Beauveria bassiana é um importe entomopatógeno, sendo avaliado seu efeito sobre a alimentação, comportamento e histologia do sistema digestório de Gallus domesticus. O fungo (isolado Unioeste 4 foi administrado oralmente às aves, misturado à ração. Estas foram observadas por 28 dias e as fezes coletadas diariamente para análise. Também foi avaliado o peso das aves, bem como sinais de intoxicação ou modificação patológica. Amostras teciduais foram retiradas para verificar lesões com o microscópico óptico. Houve sobrevivência de 100% das aves avaliadas e nenhuma alteração comportamental ou lesão externa durante o experimento. O grupo teste apresentou o maior ganho de peso (1383,9±54,4g, sendo mais acentuado a partir da segunda semana. Observou-se a presença de conídios viáveis nas fezes somente até 24 horas após a ingestão do fungo, indicando que não houve germinação nas aves. Nenhuma lesão tecidual foi verificada, de forma que B. bassiana mostrou-se seguro para o organismo n

  2. Genetic Polymorphisms and Phenotypic Profiles of Sulfadiazine-Resistant and Sensitive Toxoplasma gondii Isolates Obtained from Newborns with Congenital Toxoplasmosis in Minas Gerais, Brazil

    Science.gov (United States)

    Silva, Letícia Azevedo; Reis-Cunha, João Luís; Bartholomeu, Daniella Castanheira; Vítor, Ricardo Wagner Almeida

    2017-01-01

    Background Previous Toxoplasma gondii studies revealed that mutations in the dhps (dihydropteroate synthase) gene are associated with resistance to sulfonamides. Although Brazilian strains are genotypically different, very limited data are available regarding the susceptibility of strains obtained from human to sulfonamides. The aim of this study was to evaluate the efficacy of sulfadiazine (SDZ) against Brazilian isolates of T. gondii and verify whether isolates present polymorphisms in the dhps gene. We also investigated whether the virulence-phenotype and/or genotype were associated with the profile of susceptibility to SDZ. Methods Five T. gondii isolates obtained from newborns with congenital toxoplasmosis were used to verify susceptibility. Mice were infected with 104 tachyzoites and orally treated with different doses of SDZ. The mortality curve was evaluated by the Log-rank test. The presence of polymorphisms in the dhps gene was verified using sequencing. A descriptive analysis for 11 Brazilian isolates was used to assess the association between susceptibility, genotype, and virulence-phenotype. Results Statistical analysis showed that TgCTBr03, 07, 08, and 16 isolates were susceptible to SDZ, whereas TgCTBr11 isolate presented a profile of resistance to SDZ. Nineteen polymorphisms were identified in dhps exons. Seven polymorphisms corresponded to non-synonymous mutations, with four being new mutations, described for the first time in this study. No association was found between the profile of susceptibility and the virulence-phenotype or genotype of the parasite. Conclusions There is a high variability in the susceptibilities of Brazilian T. gondii strains to SDZ, with evidence of drug resistance. Despite the large number of polymorphisms identified, the profile of susceptibility to SDZ was not associated with any of the dhps variants identified in this study. Other genetic factors, not yet determined, may be associated with the resistance to SDZ; thus

  3. Vasorelaxant activity of extracts obtained from Apium graveolens:Possible source for vasorelaxant molecules isolation with potential antihypertensive effect

    Institute of Scientific and Technical Information of China (English)

    Vergara-Galicia Jorge; Jimenez-Ramirez Luis ngel; Tun-Suarez Adrin; Aguirre-Crespo Francisco; Salazar-Gmez Anuar; Estrada-Soto Samuel; Sierra-Ovando ngel; Hernandez-Nuez Emmanuel

    2013-01-01

    Objective:To investigate the vasorelaxant effect of organic extracts from Apium graveolens (A. graveolens) which is a part of a group of plants subjected to pharmacological and phytochemical study with the purpose of offering it as an ideal source for obtaining lead compounds for designing new therapeutic agents with potential vasorelaxant and antihypertensive effects. Methods:An ex vivo method was employed to assess the vasorelaxant activity. This consisted of using rat aortic rings with and without endothelium precontracted with norepinephrine. Results:All extracts caused concentration-dependent relaxation in precontracted aortic rings with and without endothelium;the most active extracts were Dichloromethane and Ethyl Acetate extracts from A. graveolens. These results suggested that secondary metabolites responsible for the vasorelaxant activity belong to a group of compounds of medium polarity. Also, our evidence showed that effect induced by dichloromethane and ethyl acetate extracts from A. graveolens is mediated probably by calcium antagonism. Conclusions: A. graveolens represents an ideal source for obtaining lead compounds for designing new therapeutic agents with potential vasorelaxant and antihypertensive effects.

  4. Insecticidal effects of zinc oxide nanoparticles and Beauveria bassiana TS11 on Trialeurodes vaporariorum (Westwood, 1856 (Hemiptera: Aleyrodidae

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    Zahra KHOOSHE-BAST

    2016-10-01

    Full Text Available Greenhouse whitefly, Trialeurodes vaporariorum is a major pest of horticultural and ornamental plants and is usually controlled with insecticides or biological control agents. In the current study, we examined the effects of synthesized zinc oxide nanoparticles (ZnO NPs and Beauveria bassiana TS11 on T. vaporariorum adults. ZnO NPs were synthesized by precipitation method. Field emission scanning electron microscope images indicated that ZnO NPs were non-compacted uniformly. X-ray diffraction results confirmed the hexagonal wurtzite structure of ZnO NPs. Fourier transform infrared analysis showed an intense absorption peak at a range of 434-555 cm-1 related to Zn-O bond. In bioassays, adults were exposed to different concentrations of ZnO NPs (3, 5, 10, 15, 20 mg l-1 and fungi (104, 105, 106, 107, 108 spores ml-1. LC50 values for ZnO NPs and fungi were 7.35 mg l-1and 3.28×105 spores ml-1, respectively. Mortality rates obtained with ZnO NPs and fungi at the highest concentration were 91.6 % and 88.8 %, respectively. The results indicate a positive effect of ZnO NPs and B. bassiana TS11on adults. The current study was conducted under laboratory conditions, therefore, more studies are needed in field.

  5. KDNA genetic signatures obtained by LSSP-PCR analysis of Leishmania (Leishmania infantum isolated from the new and the old world.

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    Janaína Sousa Campos Alvarenga

    Full Text Available BACKGROUND: Visceral Leishmaniasis (VL caused by species from the Leishmania donovani complex is the most severe form of the disease, lethal if untreated. VL caused by Leishmania infantum is a zoonosis with an increasing number of human cases and millions of dogs infected in the Old and the New World. In this study, L. infantum (syn. L.chagasi strains were isolated from human and canine VL cases. The strains were obtained from endemic areas from Brazil and Portugal and their genetic polymorphism was ascertained using the LSSP-PCR (Low-Stringency Single Specific Primer PCR technique for analyzing the kinetoplastid DNA (kDNA minicircles hypervariable region. PRINCIPAL FINDINGS: KDNA genetic signatures obtained by minicircle LSSP-PCR analysis of forty L. infantum strains allowed the grouping of strains in several clades. Furthermore, LSSP-PCR profiles of L. infantum subpopulations were closely related to the host origin (human or canine. To our knowledge this is the first study which used this technique to compare genetic polymorphisms among strains of L. infantum originated from both the Old and the New World. CONCLUSIONS: LSSP-PCR profiles obtained by analysis of L. infantum kDNA hypervariable region of parasites isolated from human cases and infected dogs from Brazil and Portugal exhibited a genetic correlation among isolates originated from the same reservoir, human or canine. However, no association has been detected among the kDNA signatures and the geographical origin of L. infantum strains.

  6. 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

  7. Natural variation in virulence of the entomopathogenic fungus Beauveria bassiana against malaria mosquitoes

    NARCIS (Netherlands)

    Valero Jimenez, C.A.; Debets, A.J.M.; Kan, van J.A.; Schoustra, S.E.; Takken, W.; Zwaan, B.J.; Koenraadt, C.J.M.

    2014-01-01

    Background Insecticide resistance is greatly hampering current efforts to control malaria and therefore alternative methods are needed. Entomopathogenic fungi have been proposed as an alternative with a special focus on the cosmopolitan species Beauveria bassiana. However, few studies have analysed

  8. First report of pathogenicity of Beauveria bassiana RBL1034 to the ...

    African Journals Online (AJOL)

    SERVER

    2008-04-17

    Apr 17, 2008 ... mosquitoes as well as among the different species of the genus. Anopheles. .... to both sexes of A. gambiae and Culex quinquefasciatus and mosquitoes in ... studies on the pathogenicity of the fungus Beauveria bassiana to.

  9. Developing a dual agent biopesticide formulation with Bacillus thuringiensis var. kurstaki and Beauveria bassiana blastospores

    Science.gov (United States)

    Production and formulation techniques to create a biopesticide with a mixture of entomopathogenic microbes were studied for improvements in efficacy. Recently developed production techniques were used to produce blastospores of Beauveria bassiana (Balsamo) Vuillemin strain GHA (Bb), which were mixed...

  10. KOMPATIBILITAS CENDAWAN ENTOMOPATOGEN BEAUVERIA BASSIANA (BALS VUILL (DEUTEROMYCOTINA: HYPHOMYCETES DENGAN MINYAK SERAI WANGI

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    . Trizelia

    2013-09-01

    Full Text Available Entomopathogenic fungi  such as Beauveria bassiana are important natural control agents of many insects and can be potentially used as a bioinsecticide against several pests. Other potential source of bioinsecticide is certain plants such as fragrant lemongrass oil.  The in vitro compatibility of the entomopathogenic fungus B. bassiana  and  fragrant lemongrass oil was evaluated.  Fragrant lemongrass oil was tested in three different concentrations (0.1, 0.3 and 0.5%. Fragrant lemongrass  oil  was diluted in the steril SDAY medium at the different concentrations. Effects of these concentrations on conidia germination, colony growth and sporulation were compared.  Fragrant lemongrass  oil  affected conidial germination, colony growth and sporulation of B. bassiana. Fragrant lemongrass  oil  was not compatible with the entomopathogenic fungus B.  bassiana.

  11. 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...

  12. Soil application of Beauveria bassiana GHA against apple sawfly, Hoplocampa testudinea (Hymenoptera: Tenthredinidae)

    DEFF Research Database (Denmark)

    Świergiel, Weronika; Meyling, Nicolai Vitt; Porcel, Mario;

    2016-01-01

    Low impact alternatives to synthetic insecticides for the control of apple sawfly (Hoplocampa testudinea Klug) are scarce encumbering pest management in organic apple orchards. We investigated the soil persistence and field efficacy of the entomopathogenic fungus Beauveria bassiana (Balsamo...

  13. Struktur komunitas laba-laba di ekosistem padi ratun: pengaruh aplikasi Beauveria bassiana (Balsamo

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    Siti Herlinda

    2016-02-01

    Full Text Available Spiders are the main predators in paddy ecosystem. Application of entomopathogenic fungus, such as Beauveria bassiana (Balsamo is not expected to have any negative effect on spiders in the paddy field. This research was aimed to compare species composition, abundance, and species richness of spiders in ratoon paddy. The research was done on  three hectares of rice fields that were treated with the following treatment, (i 2 L/ha/application of liquid B. bassiana, (ii 2 L/ha/application compost extract (without liquid B. bassiana, and  (iii a combination of liquid B. bassiana and compost extract with each using the same 2L/ha/application dose. The result  of this research showed that B. bassiana bioinsecticide application did not affect the abundance and species richness of spiders in ratoon paddy. The treatments did not significantly affect species richness and abundance of the canopy-inhabiting spiders. The species richness and abundance of the soil-dwelling spiders on the paddy field applied by the bioinsecticide was not different from those without the bioinsecticide. The treatments did not effect species composition of the canopy-inhabiting and the soil-dwelling spiders. Application of B. bassiana bioinsecticide was not harmful for the spiders in the rice field. 

  14. Diversity of Fungi Associated with Atta bisphaerica (Hymenoptera: Formicidae: The Activity of Aspergillus ochraceus and Beauveria bassiana

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    Myriam M. R. Ribeiro

    2012-01-01

    Full Text Available The grass-cutting ant Atta bisphaerica is one of the most serious pests in several pastures and crops in Brazil. Fungal diseases are a constant threat to these large societies composed of millions of closely related individuals. We investigated the occurrence of filamentous fungi associated with the ant A. bisphaerica in a pasture area of Viçosa, Minas Gerais State, Brazil. Several fungi species were isolated from forager ants, and two of them, known as entomopathogenic, Beauveria bassiana and Aspergillus ochraceus, were tested against worker ants in the laboratory. The two species were highly virulent, achieving 50 percent worker mortality within 4-5 days. It is the first time A. ochraceus, a commonly found fungal species, is reported to infect Atta species at a high prevalence. Possible uses for the fungus within biological control are discussed.

  15. A molecular tool for detection and tracking of a potential indigenous Beauveria bassiana strain for managing emerald ash borer populations in Canada.

    Science.gov (United States)

    Johny, Shajahan; Kyei-Poku, George

    2014-10-01

    Emerald ash borer is an invasive species from Asia. Beauveria bassiana strain L49-1AA is being tested for the control of emerald ash borer in Canada, using an autocontamination trapping system. We have developed a simplified allele discrimination polymerase chain reaction (PCR) assay to screen B. bassiana strain, L49-1AA from other Beauveria species by targeting the inter-strain genetic differences in 5' end of EF1-α gene of the genus Beauveria. A single nucleotide polymorphism (SNP) site, T→C was identified only in L49-1AA and was used to develop a simplified allele discrimination polymerase chain reaction (PCR) assay based on a modified allelic inhibition of displacement activity (AIDA) approach for distinguishing B. bassiana L49-1AA from all background Beauveria isolates. The SNP site was employed to design inner primers but with a deliberate mismatch introduced at the 3' antepenultimate from the mutation site in order to maximize specificity and detection efficiency. Amplification was specific to L49-1AA without cross-reaction with DNA from other Beauveria strains. In addition, the designed primers were also tested against environmental samples in L49-1AA released plots and observed to be highly efficient in detecting and discriminating the target strain, L49-1AA from both pure and crude DNA samples. This new method can potentially allow for more discriminatory tracking and monitoring of released L49-1AA in our autocontamination and dissemination projects for managing EAB populations. Additionally, the modified-AIDA format has potential as a tool for simultaneously identifying and differentiating closely related Beauveria species, strains/isolates as well as general classification of other pathogens or organisms.

  16. Characterization of T-DNA insertion mutants with decreased virulence in the entomopathogenic fungus Beauveria bassiana JEF-007.

    Science.gov (United States)

    Kim, Sihyeon; Lee, Se Jin; Nai, Yu-Shin; Yu, Jeong Seon; Lee, Mi Rong; Yang, Yi-Ting; Kim, Jae Su

    2016-10-01

    The bean bug, Riptortus pedestris, is a major agricultural pest that reduces crop quality and value. Chemical pesticides have contributed to pest management, but resistance to these chemicals has significantly limited their use. Alternative strategies with different modes of action, such as entomopathogenic fungi, are therefore of great interest. Herein, we explored how entomopathogenic fungi can potentially be used to control the bean bug and focused on identifying virulence-related genes. Beauveria bassiana (JEF isolates) were assayed against bean bugs under laboratory conditions. One isolate, JEF-007, showed >80 % virulence by both spray and contact exposure methods. Agrobacterium tumefaciens-mediated transformation (AtMT) of JEF-007 generated 249 random transformants, two of which (B1-06 and C1-49) showed significantly reduced virulence against Tenebrio molitor and R. pedestris immatures. Both species were used for rapid screening of virulence-reduced mutants. The two transformants had different morphologies, conidial production, and thermotolerance than the wild type. To determine the localization of the randomly inserted T-DNA, thermal asymmetric interlaced (TAIL) PCR was conducted and analysis of the two clones found multiple T-DNA insertions (two in B1-06 and three in C1-49). Genes encoding complex I intermediate-associated protein 30 (CIA30) and the autophagy protein (Atg22) were possibly disrupted by the T-DNA insertion and might be involved in the virulence. This work provides a strong platform for future functional genetic studies of bean bug-pathogenic B. bassiana. The genes putatively involved in fungal virulence should be experimentally validated by knockdown in future studies.

  17. Improving Efficacy of Beauveria bassiana against Stored Grain Beetles with a Synergistic Co-Formulant

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    Clare Storm

    2016-08-01

    Full Text Available The potential of a dry powder co-formulant, kaolin, to improve the control of storage beetles by the entomopathogenic fungus Beauveria bassiana, isolate IMI389521, was investigated. The response of Oryzaephilus surinamensis adults to the fungus when applied to wheat at 1 × 1010 conidia per kg with and without kaolin at 1.74 g per kg wheat was assessed. Addition of kaolin increased control from 46% to 88% at day 7 and from 81% to 99% at day 14 post-treatment. Following this the dose response of O. surinamensis and Tribolium confusum to both kaolin and the fungus was investigated. Synergistic effects were evident against O. surinamensis at ≥0.96 g of kaolin per kg of wheat when combined with the fungus at all concentrations tested. For T. confusum, adult mortality did not exceed 55%, however, the larvae were extremely susceptible with almost complete suppression of adult emergence at the lowest fungal rate tested even without the addition of kaolin. Finally, the dose response of Sitophilus granarius to the fungus at 15 and 25 °C, with and without kaolin at 1 g per kg of wheat, was examined. Improvements in efficacy were achieved by including kaolin at every fungal rate tested and by increasing the temperature. Kaolin by itself was not effective, only when combined with the fungus was an effect observed, indicating that kaolin was having a synergistic effect on the fungus.

  18. Improving Efficacy of Beauveria bassiana against Stored Grain Beetles with a Synergistic Co-Formulant

    Science.gov (United States)

    Storm, Clare; Scoates, Freya; Nunn, Adam; Potin, Olivier; Dillon, Aoife

    2016-01-01

    The potential of a dry powder co-formulant, kaolin, to improve the control of storage beetles by the entomopathogenic fungus Beauveria bassiana, isolate IMI389521, was investigated. The response of Oryzaephilus surinamensis adults to the fungus when applied to wheat at 1 × 1010 conidia per kg with and without kaolin at 1.74 g per kg wheat was assessed. Addition of kaolin increased control from 46% to 88% at day 7 and from 81% to 99% at day 14 post-treatment. Following this the dose response of O. surinamensis and Tribolium confusum to both kaolin and the fungus was investigated. Synergistic effects were evident against O. surinamensis at ≥0.96 g of kaolin per kg of wheat when combined with the fungus at all concentrations tested. For T. confusum, adult mortality did not exceed 55%, however, the larvae were extremely susceptible with almost complete suppression of adult emergence at the lowest fungal rate tested even without the addition of kaolin. Finally, the dose response of Sitophilus granarius to the fungus at 15 and 25 °C, with and without kaolin at 1 g per kg of wheat, was examined. Improvements in efficacy were achieved by including kaolin at every fungal rate tested and by increasing the temperature. Kaolin by itself was not effective, only when combined with the fungus was an effect observed, indicating that kaolin was having a synergistic effect on the fungus. PMID:27571107

  19. Transformation of Beauveria bassiana to produce EGFP in Tenebrio molitor for use as animal feed additives.

    Science.gov (United States)

    Kim, Jae Su; Choi, Jae Young; Lee, Se Jin; Lee, Ju Hyun; Fu, Zhenli; Skinner, Margaret; Parker, Bruce L; Je, Yeon Ho

    2013-07-01

    Efforts are underway to develop more effective and safer animal feed additives. Entomopathogenic fungi can be considered practical expression platforms of functional genes within insects which have been used as animal feed additives. In this work, as a model, the enhanced green fluorescent protein (egfp) gene was expressed in yellow mealworms, Tenebrio molitor by highly infective Beauveria bassiana ERL1170. Among seven test isolates, ERL1170 treatment showed 57.1% and 98.3% mortality of mealworms 2 and 5 days after infection, respectively. The fungal transformation vector, pABeG containing the egfp gene, was inserted into the genomic DNA of ERL1170 using the restriction enzyme-mediated integration method. This resulted in the generation of the transformant, Bb-egfp#3, which showed the highest level of fluorescence. Bb-egfp#3-treated mealworms gradually turned dark brown, and in 7-days mealworm sections showed a strong fluorescence. This did not occur in the wild-type strain. This work suggests that further valuable proteins can be efficiently produced in this mealworm-based fungal expression platform, thereby increasing the value of mealworms in the animal feed additive industry.

  20. Improving Efficacy of Beauveria bassiana against Stored Grain Beetles with a Synergistic Co-Formulant.

    Science.gov (United States)

    Storm, Clare; Scoates, Freya; Nunn, Adam; Potin, Olivier; Dillon, Aoife

    2016-08-26

    The potential of a dry powder co-formulant, kaolin, to improve the control of storage beetles by the entomopathogenic fungus Beauveria bassiana, isolate IMI389521, was investigated. The response of Oryzaephilus surinamensis adults to the fungus when applied to wheat at 1 × 10(10) conidia per kg with and without kaolin at 1.74 g per kg wheat was assessed. Addition of kaolin increased control from 46% to 88% at day 7 and from 81% to 99% at day 14 post-treatment. Following this the dose response of O. surinamensis and Tribolium confusum to both kaolin and the fungus was investigated. Synergistic effects were evident against O. surinamensis at ≥0.96 g of kaolin per kg of wheat when combined with the fungus at all concentrations tested. For T. confusum, adult mortality did not exceed 55%, however, the larvae were extremely susceptible with almost complete suppression of adult emergence at the lowest fungal rate tested even without the addition of kaolin. Finally, the dose response of Sitophilus granarius to the fungus at 15 and 25 °C, with and without kaolin at 1 g per kg of wheat, was examined. Improvements in efficacy were achieved by including kaolin at every fungal rate tested and by increasing the temperature. Kaolin by itself was not effective, only when combined with the fungus was an effect observed, indicating that kaolin was having a synergistic effect on the fungus.

  1. A Novel Approach for Ovine Primary Alveolar Epithelial Type II Cell Isolation and Culture from Fresh and Cryopreserved Tissue Obtained from Premature and Juvenile Animals.

    Science.gov (United States)

    Marcinkiewicz, Mariola M; Baker, Sandy T; Wu, Jichuan; Hubert, Terrence L; Wolfson, Marla R

    2016-01-01

    The in vivo ovine model provides a clinically relevant platform to study cardiopulmonary mechanisms and treatments of disease; however, a robust ovine primary alveolar epithelial type II (ATII) cell culture model is lacking. The objective of this study was to develop and optimize ovine lung tissue cryopreservation and primary ATII cell culture methodologies for the purposes of dissecting mechanisms at the cellular level to elucidate responses observed in vivo. To address this, we established in vitro submerged and air-liquid interface cultures of primary ovine ATII cells isolated from fresh or cryopreserved lung tissues obtained from mechanically ventilated sheep (128 days gestation-6 months of age). Presence, abundance, and mRNA expression of surfactant proteins was assessed by immunocytochemistry, Western Blot, and quantitative PCR respectively on the day of isolation, and throughout the 7 day cell culture study period. All biomarkers were significantly greater from cells isolated from fresh than cryopreserved tissue, and those cultured in air-liquid interface as compared to submerged culture conditions at all time points. Surfactant protein expression remained in the air-liquid interface culture system while that of cells cultured in the submerged system dissipated over time. Despite differences in biomarker magnitude between cells isolated from fresh and cryopreserved tissue, cells isolated from cryopreserved tissue remained metabolically active and demonstrated a similar response as cells from fresh tissue through 72 hr period of hyperoxia. These data demonstrate a cell culture methodology using fresh or cryopreserved tissue to support study of ovine primary ATII cell function and responses, to support expanded use of biobanked tissues, and to further understanding of mechanisms that contribute to in vivo function of the lung.

  2. Compatibility and survival of beneficial microorganisms for agricultural use (Beauveria bassiana, Paecilomyces lilacinus and Bacillus thuringiensis in compost

    Directory of Open Access Journals (Sweden)

    Villacís-Aldaz Luis Alfredo

    2016-11-01

    Full Text Available Compatibility and survivorship of beneficial microorganism in compost were evaluated in order to provide new alternatives to clean production in agriculture. The study was conducted at the Ecological Farm belonging to the Consejo Provincial de Tungurahua, Ecuador. Four treatments were considered related to dose of several beneficial microorganisms (EMs. After an interval of 30 or 60 days after composting process, analysis of nutritional biological content showed compatibility and survivorship of the EMs, which live together and reproduce on the organic fertilizer. Concerning physic-chemical analysis higher organic matter content, pH, electrical conductivity, nitrogen, phosphorous, potassium and calcium were evidenced in T1: 100 cc Beauveria bassiana - 100 cc Bacillus thuringiensis - 100 cc Paecilomyces lilacinus. From the results obtained will offer the possibility of using beneficial microorganisms to optimize their performance in obtaining compost of better nutritional and biological quality, so that gradually reduce the use of agrochemicals.

  3. Variation of TTC Repeat Pattern In The Dna of Mycobacterium Leprae Isolates Obtained from Archeological Bones and Leprosy Patients From East Nusa Tenggara

    Directory of Open Access Journals (Sweden)

    Dinar Adriaty

    2012-12-01

    Full Text Available The existence of leprosy or kusta or Morbus Hansen or Hansen’s disease has been known for years, including in Indonesia. Starting from the discovery of Mycobacterium leprae isolates from ancient bone (about 1.000 years B.C, the archaeological excavations results in East Nusa Tenggara, interesting questions arise about how the development of leprosy in eastern Indonesia is. Biology molecular study would become a powerful tool to investigate the presence of leprosy bacillary whether there are similarities between the genomes of M. leprae isolates in the primeval and the present. PCR examinations were performed on mandibular bone fragments from ancient human who lived 1000 years B.C. discovered in archaeological surveys on the island of Lembata and three leprosy patients from East Nusa Tenggara. The DNA extraction was performed using a kit from Qiagen products and its TTC repeating pattern was seen with the method of direct sequencing. It turned out that the TTC profile obtained from samples of archaeological was as many as 13 copies, while the repetition of TTC in three samples of leprosy patients were 15, 17 and 26 copies. The different number of TTC repetition shows the different isolates of M. leprae between in the ancient times and the present. Further studies are needed to verify the differences in the genome that occur, for example from the study of SNPs (single nucleotide polymorphisms.

  4. Prevalence and Phylogenetic Characterization of Escherichia coli and Hygiene Indicator Bacteria Isolated from Leafy Green Produce, Beef, and Pork Obtained from Farmers' Markets in Pennsylvania.

    Science.gov (United States)

    Scheinberg, Joshua A; Dudley, Edward G; Campbell, Jonathan; Roberts, Beth; DiMarzio, Michael; DebRoy, Chitrita; Cutter, Catherine N

    2017-02-01

    The popularity of farmers' markets in the United States has led to over 8,400 farmers' markets being in operation in 2015. As farmers' markets have increased in size and complexity in the kinds of foods sold at these venues, so have the potential food safety risks. Since 2008, seven major foodborne illness outbreaks and two recalls associated with food products from farmers' markets have occurred, causing 80 known reported illnesses and one death. Various researchers also have observed vendors performing high-risk food safety retail behaviors, and others have identified microbiological hazards in foods sold at farmers' markets. In this study, the presence of hygiene indicators (coliforms, fecal coliforms, Listeria spp., and Escherichia coli ) was assessed in select samples of leafy green produce and meat obtained from farmers' markets in Pennsylvania. E. coli isolates were further characterized by phylogenetic profile and virulence potential. E. coli was present in 40% (20 of 50) and 18% (9 of 50) of beef and pork samples, respectively, and in 28% (15 of 54), 29% (15 of 52), and 17% (8 of 46) of kale, lettuce, and spinach samples, respectively. Listeria spp. was found in 8% (4 of 50) of beef samples, 2% (1 of 54) of kale samples, 4% (2 of 52) of lettuce samples, and 7% (3 of 46) of spinach samples. Among the 10 Listeria spp. isolates, 3 were identified as L. monocytogenes . E. coli isolated from meat samples mainly clustered into phylogroup B1 (66%; 19 of 29), whereas produce isolates clustered into phylogroups B2 (36%; 14 of 39) and B1 (33%; 13 of 39). These E. coli isolates possessed the fimH, iroN, hlyD, and eae genes associated with extraintestinal pathogenic E. coli and Shiga toxin-producing E. coli . The high prevalence but low levels of E. coli and Listeria spp. found on both produce and meat products obtained from farmers' markets in this study strongly indicate that farmers' market vendors would benefit greatly from food safety training and increased

  5. Selection, isolation and growth kinetic study of a bacterial consortium obtained from the Potengi mangrove in the presence of crude oil

    Energy Technology Data Exchange (ETDEWEB)

    Costa, C.C.; Vaz, M.R.F.; Santos, E.S.; Macedo, G.R. [Universidade Federal do Rio Grande do Norte (UFRN), Natal, RN (Brazil). Dept. de Engenharia Quimica], E-mail: natcintia@gmail.com; Costa, J.G. da [Universidade Federal do Amazonas (UFAM), Coari, AM (Brazil). Inst. de Saude e Biotecnologia

    2011-10-15

    The selection, isolation and kinetic study of a bacterial consortium obtained from a sample of soil from the Potengi mangrove, located in the city of Natal, Rio Grande do Norte, Brazil, has been carried out using the enrichment culture technique to observe aspects such as the evaluation of main growth parameters. The kinetic study used a rotary incubator shaker at 150rpm, under 30 deg C. The bacterial consortium isolated from the estuary of the Potengi River showed a good acclimation in minimum mineral medium with 1% (v/v) of oil. The cell concentration reached 2.55 g/L at 16h of cultivation and surface tension dropped. The maximum productivity in cells obtained was of 0.3 g/L.h, the specific velocity of growth was of 0.075h{sup -1}, with a generation time (tg) of 9.24h. This study seeks to demonstrate that the consortium can be used as inoculants in biological treatments, capable of reducing the waste's degradation time. (author)

  6. An optimised protocol to isolate high-quality genomic DNA from seed tissues streamlines the workflow to obtain direct estimates of seed dispersal distances in gymnosperms.

    Science.gov (United States)

    García, C; Escribano-Ávila, G

    2016-05-01

    Genotyping of maternally derived seed tissues from georefered seeds that moved away from their source tree yield direct estimates of seed dispersal distances when the location and the genotype of the fruiting tree are available. These estimates are instrumental in forecasting the response of plant communities to drivers of global change, such as fragmentation or the expansion of invasive species. Obtaining robust assessments of seed dispersal distances requires comparing reliable multilocus genotypes of maternally derived seed tissues and fruiting trees, as previously shown for angiosperm species. However, robust estimates of seed dispersal distances based on direct methods are rare in non-model gymnosperms due to the difficulty in isolating high quality DNA from inconspicuous maternally derived seed tissues. These tissues tend to yield low DNA quantities that increase the frequency of genotyping errors. Here, we deliver a step-by-step visual protocol used to identify and isolate different seed tissues of interest for dispersal studies: embryos (2n, bi-parentally derived), seed coats (2n, maternally derived), and megagametophytes (n, maternally derived). We also provide an optimised lab protocol used to obtain multilocus genotypes from the target seed tissue. These broadly applicable protocols proved successful both in avoiding contamination among different seed tissues and providing reliable multilocus genotypes.

  7. Towards the development of an autocontamination trap system to manage populations of emerald ash borer (Coleoptera: Buprestidae) with the native entomopathogenic fungus, Beauveria bassiana.

    Science.gov (United States)

    Lyons, D Barry; Iavallée, Robert; Kyei-Poku, George; Van Frankenhuyzen, Kees; Johny, Shajahan; Guertin, Claude; Francese, Joseph A; Jones, Gene C; Blais, Martine

    2012-12-01

    Emerald ash borer, Agrilus planipennis Fairmaire (Coleoptera: Buprestidae) is an invasive species from Asia that was discovered in North America Canada, in 2002. Herein, we describe studies to develop an autocontamination trapping system to disseminate Beauveria bassiana to control beetle populations. The standard trap for emerald ash borer in Canada is a light green prism trap covered in an insect adhesive and baited with (Z)-3-hexenol. We compared of green multifunnel traps, green intercept panel traps (both with and without fluon coating) and green prism traps for capturing emerald ash borer in a green ash plantation. The coated green multifunnel traps captured significantly more males and more females than any other trap design. We examined the efficacy of two native B. bassiana isolates, INRS-CFL and L49-1AA. In a field experiment the INRS-CFL isolate attached to multifunnel traps in autocontamination chambers retained its pathogenicity to emerald ash borer adults for up to 43 d of outdoor exposure. Conidia germination of the INRS-CFL isolate was >69% after outdoor exposure in the traps for up to 57 d. The L49-1AA isolate was not pathogenic in simulated trap exposures and the germination rate was extremely low (<5.3%). Mean (+/- SEM) conidia loads on ash borer adults after being autocontaminated in the laboratory using pouches that had been exposed in traps out of doors for 29 d were 579,200 (+/- 86,181) and 2,400 (+/- 681) for the INRS-CFL and the LA9-1AA isolates, respectively. We also examined the fungal dissemination process under field conditions using the L49-1AA isolate in a green ash plantation. Beetles were lured to baited green multifunnel traps with attached autocontamination chambers. Beetles acquired fungal conidia from cultures growing on pouches in the chambers and were recaptured on Pestick-coated traps. In total, 2,532 beetles were captured of which 165 (6.5%) had fungal growth that resembled B. bassiana. Of these 25 beetles were positive for

  8. Glucose concentration alters dissolved oxygen levels in liquid cultures of Beauveria bassiana and affects formation and bioefficacy of blastospores

    Science.gov (United States)

    Beauveria bassiana is a ubiquitous dimorphic entomopathogenic fungus commonly used for controlling numerous insect pests worldwide. The goal of the present study was to optimize the production of B. bassiana to achieve high yields of the preferred morphology, a yeast-like blastospore, rather than my...

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

    Directory of Open Access Journals (Sweden)

    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.

  10. Production of Beauveria bassiana Air Conidia by Means of Optimization of Biphasic System Technology

    Directory of Open Access Journals (Sweden)

    Vladimir Gouli

    2014-08-01

    Full Text Available Air conidia production of Beauveria bassiana, strain CA-603 was studied based on modified diphasic system. The biomass yield obtained in the first phase based on submerged cultivation of fungus was processed using methodology providing different contact with air space. Our study indicated that productivity of the second stage of diphasic system is had inversely proportional dependence on depth of liquid fungal biomass. Increase of biomass depth is significantly decreased production of air conidia. Two methodology of biomass processing extending contact biomass with air space including distribution of fungal material on surface of hygroscopic paper and starch packaging peanuts were investigated. The novel substrates provided optimal contact between the submerged fungal biomass and the air, and overall, conidial production was directly proportional to the total area of air-to-fungal surface. Technologies based on the starch peanuts and hygroscopic paper were the most productive in comparison to the common technology where the submerged culture was transferred to flat containers. The advantages and disadvantages of these different production methods are discussed.

  11. 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.

  12. Compatibility of Beauveria bassiana with Neoseiulus barkeri for Control of Frankliniella occidentalis

    Institute of Scientific and Technical Information of China (English)

    WU Sheng-yong; GAO Yu-lin; XU Xue-nong; Mark S Goettel; LEI Zhong-ren

    2015-01-01

    The entomopathogenic fungusBeauveriabassiana (Balsamo) Vuilemin and predatory miteNeoseiulusbarkeri Hughes are effective biological control agents ofFranklinielaoccidentalis Pergande (Thysanoptera: Thripidae), one of the most important pests of ornamentals and vegetables world-wide. Combined application of both may enhance control efifciency. The functional response forN.barkeri on the ifrst instar larvae of western lfower thrips which were infected byB.bassiana for 12 and 24 h in the laboratory ((25±1)°C, (70±5)% RH, L:D=16 h:8 h) was determined. The virulence ofB.bassiana against the second instar and pupae of the thrips attacked byN.barkeri were also tested. The results showed that N.barkeri exhibited a Holing type II functional response on treated thrips. After having been treated with the fungus for 12 h and then offered to the predator, thrips were more vulnerable to be kiled byN.barkeri. The second instar larvae and pupae of the thrips which had been attacked by predatory mites were markedly more susceptible toB.bassiana infection than normal thrips; the cumulative corrected mortality ofB.bassiana of the second instar and pupae which were attacked byN.barkeri were 57 and 94%, respectively, compared to 35 and 80% in controls on the day 8. These ifndings highlight the potential use ofB.bassiana in combination withN.barkeri to controlF. occidentalis.

  13. Increasing metronidazole and rifampicin resistance of Helicobacter pylori isolates obtained from children and adolescents between 2002 and 2015 in southwest Germany.

    Science.gov (United States)

    Regnath, Thomas; Raecke, Olaf; Enninger, Axel; Ignatius, Ralf

    2017-02-01

    Increasing antibiotic resistance has been reported for Helicobacter pylori, but data on the prevalence of antibiotic resistance of H. pylori in pediatric patients and the development of resistance over time are sparse. Data for 610 H. pylori isolates obtained between 2002 and 2015 from gastric biopsies of 582 (mainly treatment-naïve) pediatric patients from southwest Germany were analyzed retrospectively regarding the antibiotic susceptibility determined by Etest and patients' characteristics. Overall resistance to metronidazole, clarithromycin, and rifampicin was 28.7%, 23.2%, and 13.3%, respectively, while resistance to amoxicillin was rare (0.8%). Simultaneous resistance to metronidazole and clarithromycin was observed for 7.7% of the isolates, and 2.3% were resistant to metronidazole, clarithromycin, and rifampicin. Differences between primary vs secondary resistance existed for metronidazole (24.7% vs 38.8%, P=.01) and clarithromycin (17.2% vs 54.1%, P=.0001). From 2002-2008 to 2009-2015, resistance to metronidazole increased from 20.8% to 34.4% (P=.003) and to rifampicin from 3.9% to 18.8% (P=.0001); this was not associated with increased numbers of patients previously treated for H. pylori infection in the second study period. In contrast, resistance to clarithromycin did not change significantly over time. Resistance was not associated with age, sex, or family origin in Europe. The considerable antibiotic resistance of H. pylori isolates argues for standard antibiotic susceptibility testing of H. pylori in pediatric patients prior to the initiation of antibiotic therapy. © 2016 John Wiley & Sons Ltd.

  14. Role of integrons, plasmids and SXT elements in multidrug resistance of Vibrio cholerae and Providencia vermicola obtained from a clinical isolate of diarrhea

    Directory of Open Access Journals (Sweden)

    Neha eRajpara

    2015-02-01

    Full Text Available The isolates of Vibrio cholerae and Providencia vermicola obtained from a diarrhoeal patient were investigated for genetic elements governing their drug resistance phenotypes. Out of fourteen antibiotics tested, V. cholerae Vc IDH02365 isolate showed resistance to nine antibiotics, while P. vermicola Pv NBA2365 was found to be resistant to all the antibiotics except polymyxin B. Though SXT integrase was depicted in both the bacteria, class 1 integron was found to be associated only with Pv NBA2365. Integrons in Pv NBA2365 conferred resistance to β-lactams, aminoglycosides and trimethoprim. Pv NBA2365 carried two transformable plasmids imparting distinct antibiotic resistance traits to their Escherichia coli transformants. In rabbit ileal loop assays, Pv NBA2365 did not show any fluid accumulation in contrast with Vc IDH02365 that showed high fluid accumulation. To the best of our knowledge, this is the first report of a highly drug resistant P.vermicola and additionally co-existence of multidrug resistant V. cholerae and P. vermicola. Both the microbes appeared to possess a wide array of mobile genetic elements for a large spectrum of antimicrobial agents, some of which are being used in the treatment of acute diarrhoea.

  15. The infectivity of the entomopathogenic fungus Beauveria bassiana to insecticide-resistant and susceptible Anopheles arabiensis mosquitoes at two different temperatures

    Directory of Open Access Journals (Sweden)

    Koekemoer Lizette L

    2010-03-01

    C. Even so, because malaria parasite development is also known to slow as temperatures fall, expected reductions in malaria transmission potential due to fungal infection under the cooler conditions would still be high. Conclusions These results provide evidence that the entomopathogenic fungus B. bassiana has potential for use as an alternative vector control tool against insecticide-resistant mosquitoes under conditions typical of indoor resting environments. Nonetheless, the observed variation in effective virulence reveals the need for further study to optimize selection of isolates, dose and use strategy in different eco-epidemiological settings.

  16. Molecular Fingerprinting of Mycobacterium tuberculosis Isolates Obtained in Havana, Cuba, by IS6110 Restriction Fragment Length Polymorphism Analysis and by the Double-Repetitive-Element PCR Method

    Science.gov (United States)

    Montoro, Ernesto; Valdivia, José; Leão, Sylvia Cardoso

    1998-01-01

    Mycobacterium tuberculosis sputum isolates from 38 patients, obtained in the first 6 months of 1997 in Havana, Cuba, were characterized by IS6110 restriction fragment length polymorphism (RFLP) analysis and the double-repetitive-element PCR (DRE-PCR) method. Among 41 strains from 38 patients, 24 and 25 unique patterns, and 5 and 4 cluster patterns, were found by the RFLP and DRE-PCR methods, respectively. Patients within two of these clusters were found to be epidemiologically related, while no relation was observed in patients in the other clusters. The DRE-PCR method is rapid, and it was as discriminating as IS6110 RFLP analysis in identifying an epidemiological association. Its simplicity makes the technique accessible for subtyping of M. tuberculosis strains in laboratories not equipped to perform RFLP analysis. PMID:9738082

  17. Phenotypic and genotypic characters of isolates of Pasteurella multocida obtained from back-yard poultry and from two outbreaks of avian cholera in avifauna in Denmark

    DEFF Research Database (Denmark)

    Christensen, J.P.; Dietz, Hans-Henrik; Bisgaard, M.

    1998-01-01

    Two outbreaks of fowl cholera in the avifauna in Denmark, affecting primarily elders but also cormorants, gulls and oyster-catchers were shown to be caused by the same clone of Pasteurella multocida ssp, multocida by restriction enzyme analysis (REA) and ribotyping, using the enzymes HpaII and Hha......I and phenotypic characterization. This observation indicated spread by migratory birds. It was shown that the outbreak clone was closely related to isolates of Pasteurella multocida ssp, multocida obtained from back-yard poultry in Denmark, including chickens, pheasants, turkeys and ducks. The only detectable...... difference between the outbreak clone and some of these strains concerned the size of one fragment. These results indicate a possible exchange of P. multocida ssp, multocida between populations of wild birds and back-yard poultry. Among the DNA fingerprinting methods used, restriction enzyme analysis offered...

  18. Vigilance for Salmonella in Feedstuffs Available in Costa Rica: Prevalence, Serotyping and Tetracycline Resistance of Isolates Obtained from 2009 to 2014.

    Science.gov (United States)

    Molina, Andrea; Granados-Chinchilla, Fabio; Jiménez, Marisol; Acuña-Calvo, María Teresa; Alfaro, Margarita; Chavarría, Guadalupe

    2016-03-01

    Relevant epidemiological information is provided in this report for Salmonella based on data obtained from a Costa Rican surveillance program for animal feeds. In addition to prevalence, a description in terms of serotypes and tetracycline (TET) resistance of the isolates is included. A total of 1725 feed and feed ingredients samples were analyzed during 2009 and 2014, from which 110 Salmonella strains were recovered (76 from poultry, 23 from meat and bone meal [MBM], 3 from pet foods, and 8 from other feed). Retrieved isolates were serotyped and tested for minimum inhibitory concentration (MIC) against TET. Salmonella strains were found mainly from poultry feed (different growth stages, n = 76/110; 69.1%) and MBM (n = 23/109; 21.1%). The rest of the isolates were recovered from feather meal, pet food, fish meal (n = 3/110; 2.3% each) and swine feed (n = 1/110; 0.9%). From the different serotypes recovered (n = 21), the most common were Salmonella Give (n = 18; 13.8%) and Salmonella Rissen (n = 6; 4.6%) for MBM and Salmonella Havana (n = 14; 10.8%), Salmonella Rissen, Salmonella Soerenga, and Salmonella Schwarzengrund (n = 8; 6.2% each) in poultry feed. Recovered strains were regarded to be sensitive or have an intermediate resistance to TET as evidenced by their MIC50 and MIC90 concentrations of 4 and 8 μg/mL for MBM and poultry feed, respectively. Compound feed and MBM samples exhibited strains characterized by 86.8 and 88.9% of the isolates classified (according to CLSI, 2015 ) as sensitive, 7.7 and 3.7% as intermediate, and 5.5% (with >256 μg/mL as the highest concentration) and 7.4% (with 64 μg/mL as the highest concentration) as resistant to TET, respectively. Salmonella serovars Anatum and Havana exhibited the highest resistance profile >256 and 128 μg/mL, respectively. Hence, MBM and poultry feed seem to be a target of interest if Salmonella incidence is to be controlled. Serotypes recovered have in the past

  19. PATOGENITAS CENDAWAN BEAUVERIA BASSIANA TERHADAP LARVA NYAMUK AEDES AEGYPTI DAN CULEX PIPIENS QUINQUEFASCIATUS DI LABORATORIUM

    Directory of Open Access Journals (Sweden)

    Amrul Munif

    2012-09-01

    Full Text Available The capability of Beauveria bassiana fungus to kill mosquito larvae was challenged with Aedes aegypti and Culex pipiens quinquefasciatus in a study conducted at the entomology laboratory of the Health Ecology Research Centre. Cx. p. quinquefeasciatus was more sensitive compared to Ae. aegypti to the B. bassiana strain from Sukamandi (West Java, which is probably due to the mosquitoes behaviour and conidiospore larvacidal effect. Conidia dust application, with a dosage of 2.2 mglliter, to water surface, within 48 hours was able to kill almost all the Cx. p. quinquefasciatus. However a dosage of 4 mglliter was required to kill all the Ae.aegypti. And a dosage of 1.3 mg conidiospore I liter is able to kill 50% Cx. p. quinquefasciatus. It seems that B. bassiana has greater capability to eradicate Cx. p. quinquefasciatus compared to Ae. aegypti

  20. Microbial Baeyer-Villiger oxidation of 5α-steroids using Beauveria bassiana. A stereochemical requirement for the 11α-hydroxylation and the lactonization pathway.

    Science.gov (United States)

    Świzdor, Alina; Panek, Anna; Milecka-Tronina, Natalia

    2014-04-01

    Beauveria bassiana KCH 1065, as was recently demonstrated, is unusual amongst fungal biocatalysts in that it converts C19 3-oxo-4-ene and 3β-hydroxy-5-ene as well as 3β-hydroxy-5α-saturated steroids to 11α-hydroxy ring-D lactones. The Baeyer-Villiger monooxygenase (BVMO) of this strain is distinguished from other enzymes catalyzing BVO of steroidal ketones by the fact that it oxidizes solely substrates with 11α-hydroxyl group. The current study using a series of 5α-saturated steroids (androsterone, 3α-androstanediol and androstanedione) has highlighted that a small change of the steroid structure can result in significant differences of the metabolic fate. It was found that the 3α-stereochemistry of hydroxyl group restricted "normal" binding orientation of the substrate within 11α-hydroxylase and, as a result, androsterone and 3α-androstanediol were converted into a mixture of 7β-, 11α- and 7α-hydroxy derivatives. Hydroxylation of androstanedione occurred only at the 11α-position, indicating that the 3-oxo group limits the alternative binding orientation of the substrate within the hydroxylase. Only androstanedione and 3α-androstanediol were metabolized to hydroxylactones. The study uniquely demonstrated preference for oxidation of equatorial (11α-, 7β-) hydroxyketones by BVMO from B. bassiana. The time course experiments suggested that the activity of 17β-HSD is a factor determining the amount of produced ring-D lactones. The obtained 11α-hydroxylactones underwent further transformations (oxy-red reactions) at C-3. During conversion of androstanedione, a minor dehydrogenation pathway was observed with generation of 11α,17β-dihydroxy-5α-androst-1-en-3-one. The introduction of C1C2 double bond has been recorded in B. bassiana for the first time.

  1. 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

  2. An entomopathogenic strain of Beauveria bassiana against Frankliniella occidentalis with no detrimental effect on the predatory mite Neoseiulus barkeri: evidence from laboratory bioassay and scanning electron microscopic observation.

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    Shengyong Wu

    Full Text Available Among 28 isolates of Beauveria bassiana tested for virulence against F. occidentalis in laboratory bioassays, we found strain SZ-26 as the most potent, causing 96% mortality in adults at 1×10(7 mL(-1conidia after 4 days. The effect of the strain SZ-26 on survival, longevity and fecundity of the predatory mite Neoseiulus (Amblyseius barkeri Hughes were studied under laboratory conditions. The bioassay results showed that the corrected mortalities were less than 4 and 8% at 10 days following inoculation of the adult and the larvae of the predator, respectively, with 1×10(7 conidia mL(-1 of SZ-26. Furthermore, no fungal hyphae were found in dead predators. The oviposition and postoviposition durations, longevity, and fecundity displayed no significant differences after inoculation with SZ-26 using first-instar larvae of F. occidentalis as prey in comparison with untreated predator. In contrast, the preoviposition durations were significantly longer. Observations with a scanning electron microscope, revealed that many conidia were attached to the cuticles of F. occidentalis at 2 h after treatment with germ tubes oriented toward cuticle at 24 h, penetration of the insect cuticle at 36 h, and finally, fungal colonization of the whole insect body at 60 h. In contrast, we never observed penetration of the predator's cuticle and conidia were shed gradually from the body, further demonstrating that B. bassiana strain SZ-26 show high toxicity against F. occidentalis but no pathogenicity to predatory mite.

  3. Effect of Storage Temperature on Beauveria bassiana (Bals. Vuill. Viability on Several Carriers

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    Sri Sukamto

    2006-05-01

    Full Text Available One of the entomopatogenous fungus types commonly observed and showed potency as myco-insectiside is Beauveria bassiana(Bals. Vuill.In order to support effectiveness and patogenous activity of B. bassiana, it is necessary to add a carrying agent that protects its spores from ultra violet ray. This study aims to investigate the effect of storage temperature on viability of B. Bassianaspores on the carrier material. The observation was carried out in the Laboratory of Plant Diseases, Indonesian Coffee and Cocoa Research Institute. The research was arranged in completely randomized design by three factors. The first factor was carrier (C, that consists of C1 = rice flour, C2 = maize flour and C3 = tapioca flour. The second factor was dosage (D, that consists of D1 = 1 g B. bassiana+ 1 g carrier; D2 = 1 g B. bassiana+ 5 g carrier and D3 = 1 g B. bassiana+ 10 g carrier. The third factor was temperature of the storage (T,that consists of T1 = 5oC; T2 = 23oC and T3 = 29oC. Viability of B. Bassiana spores was examined by observing development of 100 blastopores randomly and determined under light microscope with 400 times magnification. Observation was conducted in two replicates after the spores of B. bassiana were kept in the storage for 2, 4, 8 and 16 weeks. The result showed that by adding 1 g tapioca flour and temperature of storage of 5oC was potentiall in keeping viability of B. bassianaspores at least for 2 months. It was due to that tapioca flour gave better effect than rice and maize flours in keeping the storage and appropriate low temperature. Viability of B. bassianaspores decreased with increasing carrier dosage, temperature and duration of the storage. Whereas, storage at 5oC was found to be a better condition in keeping viability of dry pure B. bassianaspores longer than conditions of 23o and 29oC. Key word:Beauveria bassiana, temperature, viability,carrier.

  4. In vivo expression of genes in the entomopathogenic fungus Beauveria bassiana during infection of lepidopteran larvae.

    Science.gov (United States)

    Galidevara, Sandhya; Reineke, Annette; Koduru, Uma Devi

    2016-05-01

    The entomopathogenic fungus Beauveria bassiana (Bals.) Vuillemin is commercially available as a bio insecticide. The expression of three genes previously identified to have a role in pathogenicity in in vitro studies was validated in vivo in three lepidopteran insects infected with B. bassiana. Expression of all three genes was observed in all the tested insects starting from 48 or 72h to 10d post infection corroborating their role in pathogenicity. We suggest that it is essential to test the expression of putative pathogenicity genes both in vitro and in vivo to understand their role in different insect species.

  5. [Effects of Beauveria bassiana on Myzus persicae and its two predaceous natural enemies].

    Science.gov (United States)

    Zhu, Hong; Luo, Xu-mei; Song, Jin-xin; Fan, Mei-zhen; Li, Zeng-zhi

    2011-09-01

    A Beauveria bassiana strain Bb21 was isolated from naturally infected green peach aphid Myzus persicae (Hemiptera: Aphididae). The effects of the strain on M. persicae and its two predaceous natural enemies Chrysoperla carnea (Neuroptera: Chrysopidae) and Harmonia axyridis (Coleoptera: Coccinellidae) were investigated under laboratory conditions. Bb21 had strong pathogenicity to M. persicae, with the LD50 of 97 conidia x mm(-2) (45-191, 95% confidence interval), but was less pathogenic to the second instar nymph of C. carnea, with the LD50 of 1089 conidia x mm(-2). The LD50 for C. carnea was 10.2 times higher than that for M. persicae. The pathogenicity of Bb21 to H. axyridis was very weak, with a low infection rate of 13% even at a high concentration 5 x 10(8) conidia x mL(-1). The Bb21 at low conidia concentration had less effect on the developmental period and fecundity of the two predaceous natural enemies. However, when applied at the high concentration 5 x 10(8) spores x mL(-1), Bb21 shortened the larval stage of H. axyridis averagely by 1.4 d and decreased the adult emergence rate and fecundity by 33% and 14%, respectively, and shortened the larval stage of C. carnea averagely by 0.7 d and decreased the adult emergence rate and fecundity by 24% and 11%, respectively. Since the LD50 for green peach aphid was much lower than that for the two predaceous natural enemies, and had very low effect on the adult emergence rate and fecundity of the two predators at the concentration recommended for field spray, Bb21 could be applied as a biocontrol agent of M. persicae in the integrated management of pernicious organisms.

  6. RNA-Seq Analyses for Two Silkworm Strains Reveals Insight into Their Susceptibility and Resistance to Beauveria bassiana Infection.

    Science.gov (United States)

    Xing, Dongxu; Yang, Qiong; Jiang, Liang; Li, Qingrong; Xiao, Yang; Ye, Mingqiang; Xia, Qingyou

    2017-02-10

    The silkworm Bombyx mori is an economically important species. White muscardine caused by Beauveria bassiana is the main fungal disease in sericulture, and understanding the silkworm responses to B. bassiana infection is of particular interest. Herein, we investigated the molecular mechanisms underlying these responses in two silkworm strains Haoyue (HY, sensitive to B. bassiana) and Kang 8 (K8, resistant to B. bassiana) using an RNA-seq approach. For each strain, three biological replicates for immersion treatment, two replicates for injection treatment and three untreated controls were collected to generate 16 libraries for sequencing. Differentially expressed genes (DEGs) between treated samples and untreated controls, and between the two silkworm strains, were identified. DEGs and the enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of the two strains exhibited an obvious difference. Several genes encoding cuticle proteins, serine proteinase inhibitors (SPI) and antimicrobial peptides (AMP) and the drug metabolism pathway involved in toxin detoxification were considered to be related to the resistance of K8 to B. bassiana. These results revealed insight into the resistance and susceptibility of two silkworm strains against B. bassiana infection and provided a roadmap for silkworm molecular breeding to enhance its resistance to B. bassiana.

  7. Influence of pH on viscoelastic properties of heat-induced gels obtained with a β-Lactoglobulin fraction isolated from bovine milk whey hydrolysates.

    Science.gov (United States)

    Estévez, Natalia; Fuciños, Pablo; Bargiela, Verónica; Picó, Guillermo; Valetti, Nadia Woitovich; Tovar, Clara Asunción; Rúa, M Luisa

    2017-03-15

    A β-Lactoglobulin fraction (r-βLg) was isolated from whey hydrolysates produced with cardosins from Cynara cardunculus. The impact of the hydrolysis process on the r-βLg structure and the rheological properties of heat-induced gels obtained thereafter were studied at different pH values. Differences were observed between r-βLg and commercial β-Lg used as control. Higher values for the fluorescence emission intensity and red shifts of the emission wavelength of r-βLg suggested changes in its tertiary structure and more solvent-exposed tryptophan residues. Circular dichroism spectra also supported these evidences indicating that hydrolysis yielded an intermediate (non-native) β-Lg state. The thermal history of r-βLg through the new adopted conformation improved the microstructure of the gels at acidic pH. So, a new microstructure with better rheological characteristics (higher conformational flexibility and lower rigidity) and greater water holding ability was founded for r-βLg gel. These results were reflected in the microstructural analysis by scanning electron microscopy.

  8. Genetic profiles of fluoroquinolone-resistant Escherichia coli isolates obtained from patients with cystitis: phylogeny, virulence factors, PAIusp subtypes, and mutation patterns.

    Science.gov (United States)

    Takahashi, Akira; Muratani, Tetsuro; Yasuda, Mitsuru; Takahashi, Satoshi; Monden, Koichi; Ishikawa, Kiyohito; Kiyota, Hiroshi; Arakawa, Soichi; Matsumoto, Tetsuro; Shima, Hiroki; Kurazono, Hisao; Yamamoto, Shingo

    2009-03-01

    The low virulence of quinolone- and fluoroquinolone-resistant Escherichia coli strains is known, although the reasons for this remain unclear. We surveyed the mutation patterns of quinolone resistance determining regions (QRDRs), phylogenetic distribution, prevalence of 18 urovirulence genes, and PAIusp subtypes in 89 fluoroquinolone-resistant E. coli (FQREC) isolates obtained from patients with cystitis and compared them with those of their fluoroquinolone-susceptible counterparts (FQSEC). Phylogenetic group B2 was significantly less prevalent in FQREC than in FQSEC (49% versus 78%; P=0.0138), but it still dominated, followed by phylogroup D (35%), in FQREC. When the prevalences of virulence factor (VF) genes were compared between FQREC and FQSEC, sfa/foc, cnf1, hly, kpsMT, ompT, ibeA, usp, and iroN showed significantly lower prevalences in FQREC than in FQSEC (1.1% versus 24% [Presistance, e.g., mutations in QRDRs, might be a specific event in limited strains, such as those that possess PAIusp subtype 2a in phylogroup B2.

  9. 球孢白僵菌对月季长管蚜的生物防治效果%Studies on biological control of Macrosiphum roswomm by using Beauveria bassiana

    Institute of Scientific and Technical Information of China (English)

    姚晓洁; 朱虹

    2015-01-01

    Macrosiphum roswomm is one of the most serious pests on Rosaceae. However, biological control of M. ros⁃womm with Beauveria bassiana still remains unreported. The patho genicity and greenhouse control effect of B. bassiana strain isolated from M.roswomm on aphid nymphs and adults, and the survival of B. bassiana on rose leaves were studied. The results showed that, under the same conditions, aphid mortality increased with increasing spore concentration. At the spore concentration of 108 spores/mL, the highest cumulative mortality was 99�3% in 5 days; the pathogenicity for adults and the third instar nymphs was much higher than the first instar nymph. Under different temperature and humidi⁃ty, the highest mortality was discovered at the condition of 25℃, and the infection mortality rate increased with the in⁃creasing of the environment humidity;the highest infection mortality in greenhouse was 94%. These results indicated that B.bassiana has a good potential in biological control of M. roswomm.%研究了1株分离自月季长管蚜( Macrosiphum roswomm Zhang)的球孢白僵菌( Beauveria bassiana ( Bals.) Vuill)对月季长管蚜若虫和成虫的致病力及温室防治效果,并对球孢白僵菌孢子在月季叶片上的存活动态进行了深入观测。结果表明:相同条件下,月季长管蚜死亡率随球孢白僵菌孢子浓度的增加而上升,在孢子浓度为108 spores/mL时,月季长管蚜5 d的最高累积死亡率为99.3%;球孢白僵菌对成虫和3龄若虫的致病性最强,1龄若虫最低。试验发现25℃时月季长管蚜死亡率最高,环境湿度越大,月季长管蚜的死亡率越高;温室内最高感染死亡率达94%,表明球孢白僵菌在月季长管蚜的生物防治中具有很好的应用潜力。

  10. 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...

  11. 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...

  12. Improved shelf life of dried Beauveria bassiana blastospores using convective drying and active packaging processes

    Science.gov (United States)

    The yeast form (blastospore) of the dimorphic insect-pathogenic fungus Beauveria bassiana can be rapidly produced using liquid fermentation methods but is generally unable to survive rapid dehydration processes or storage under non-refrigerated conditions. In this study, we evaluated the influence o...

  13. 75 FR 10186 - Beauveria bassiana HF23; Amendment of Exemption from the Requirement of a Tolerance

    Science.gov (United States)

    2010-03-05

    ... [Federal Register Volume 75, Number 43 (Friday, March 5, 2010)] [Rules and Regulations] [Pages 10186-10190] [FR Doc No: 2010-4544] ENVIRONMENTAL PROTECTION AGENCY 40 CFR Part 180 [EPA-HQ-OPP-2005-0316; FRL-8814-6] Beauveria bassiana HF23; Amendment of Exemption from the Requirement of a...

  14. Cordyceps bassiana and production of stromata in vitro showing Beauveria anamorph in Korea

    Science.gov (United States)

    A Cordyceps species was found with Beauveria anamorph state on larval insect cadavers on Obong Mountsin in Gangwon Pronvince, Republic of Korea. Cultures from discharged ascospores formed an anamorph identifiable as Beauveria bassiana. This teleomorph-anamorph connection was also confirmed by the in...

  15. 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.

  16. Regulatory cascade and biological activity of Beauveria bassiana oosporein that limits bacterial growth after host death.

    Science.gov (United States)

    Fan, Yanhua; Liu, Xi; Keyhani, Nemat O; Tang, Guirong; Pei, Yan; Zhang, Wenwen; Tong, Sheng

    2017-02-28

    The regulatory network and biological functions of the fungal secondary metabolite oosporein have remained obscure. Beauveria bassiana has evolved the ability to parasitize insects and outcompete microbial challengers for assimilation of host nutrients. A novel zinc finger transcription factor, BbSmr1 (B. bassiana secondary metabolite regulator 1), was identified in a screen for oosporein overproduction. Deletion of Bbsmr1 resulted in up-regulation of the oosporein biosynthetic gene cluster (OpS genes) and constitutive oosporein production. Oosporein production was abolished in double mutants of Bbsmr1 and a second transcription factor, OpS3, within the oosporein gene cluster (ΔBbsmr1ΔOpS3), indicating that BbSmr1 acts as a negative regulator of OpS3 expression. Real-time quantitative PCR and a GFP promoter fusion construct of OpS1, the oosporein polyketide synthase, indicated that OpS1 is expressed mainly in insect cadavers at 24-48 h after death. Bacterial colony analysis in B. bassiana-infected insect hosts revealed increasing counts until host death, with a dramatic decrease (∼90%) after death that correlated with oosporein production. In vitro studies verified the inhibitory activity of oosporein against bacteria derived from insect cadavers. These results suggest that oosporein acts as an antimicrobial compound to limit microbial competition on B. bassiana-killed hosts, allowing the fungus to maximally use host nutrients to grow and sporulate on infected cadavers.

  17. Efficacy of Molasses and Beauveria bassiana (Balsamo Vuill on the Control of Cosmopolites sorditus Germar, 1824

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    Flávia Queiroz de Oliveira

    2010-06-01

    Full Text Available This research sought to study the compatibility between the molasses and the entomopathogenic fungi Beauveria bassiana (Balsamo Vuill on the control of Cosmopolites sorditus Germar, 1824. The study was conducted on the Site Pitiá in a banana plantation of approximately 250 m2, located in the municipality of Areia, state of Paraiba, Brazil. The results showed that the constancy of C. sorditus was significantly higher in traps consisting of molasses+fungi (P <0.01. And significantly lower in traps without yeast and molasses. The frequency of C. sorditus was classified as accidental in traps with only B. bassiana (23.44% and in traps without the addition of B. bassiana and molasses (10.71%, and being accessory in traps consisting of molasses (28.43% and in traps with molasses+fungi (37.27%. Therefore the food bait molasses along with the fungi B. bassiana, may be used in an integrated control of C. sorditus in banana plantations.

  18. Calmodulin-mediated suppression of 2-ketoisovalerate reductase in Beauveria bassiana beauvericin biosynthetic pathway.

    Science.gov (United States)

    Kim, Jiyoung; Yoon, Deok-Hyo; Oh, Junsang; Hyun, Min-Woo; Han, Jae-Gu; Sung, Gi-Ho

    2016-11-01

    Ketoisovalerate reductase (KIVR, E.C. 1.2.7.7) mediates the specific reduction of 2-ketoisovalerate (2-Kiv) to d-hydroxyisovalerate (d-Hiv), a precursor for beauvericin biosynthesis. Beauvericin, a famous mycotoxin produced by many fungi, is a cyclooligomer depsipeptide, which has insecticidal, antimicrobial, antiviral, and cytotoxic activities. In this report, we demonstrated that Beauveria bassiana 2-ketoisovalerate reductase (BbKIVR) acts as a typical KIVR enzyme in the entomopathogenic fungus B. bassiana. In addition, we found that BbKIVR interacts with calmodulin (CaM) in vitro and in vivo. The functional role of CaM-binding to BbKIVR was to negatively regulate the BbKIVR activity in B. bassiana. Environmental stimuli such as light and salt stress suppressed BbKIVR activity in B. bassiana. Interestingly, this negative effect of BbKIVR activity by light and salt stress was recovered by CaM inhibitors, suggesting that the inhibitory mechanism is mediated through stimulation of CaM activity. Therefore, this work suggests that BbKIVR plays an important role in the beauvericin biosynthetic pathway mediated by environmental stimuli such as light and salt stress via the CaM signaling pathway.

  19. 生物防治白僵菌与家蚕病原白僵菌的生物学特性初步比较%A Preliminary Comparison on Biological Characteristics of Biocontrol Agent Beauveria bassiana and Silkworm Pathogen Beauveria bassiana

    Institute of Scientific and Technical Information of China (English)

    刘吉平; 吕思行; 米红霞

    2011-01-01

    In order to provide experimental data for the source-tracking and control of white muscardine disease and for safety assessment of applying Beauveria bassiana insecticide in silkworm rearing areas, PDA medium and scanning electron microscopy were used for isolation, identification and observation of 2 biocontrol agents, namely B. bassiana strain Bb2 used for forest protection in Guangzhou City and B. bassiana strain Bb7 preserved in the South China Agricultural University, and of 3 B. bassiana isolates collected in Guangdong provincial silkworm rearing areas at Huazhou Silkworm Eggs Production Farm, Daqiao Village of Huazhou City, and Xiao Village of Wengyuan County, respectively. A prelimina ry comparison was conducted on the biological characteristics between the above5 strains. The results showed that there existed differences on the colonial morphology, vegetative growth, spore quantity and other biological characteristics of the 5 strains. Among them, strain Bb2 grew the fastest, and strains Bb8 and Bb13 had the highest spore quantities. The conidiospores from muscardine pupae infected by strains Bb8 and Bb14 were spherical or near-spherical under scanning electron microscope. Both of them belonged to [ Beauveria bassiana (Bals.) Vuill. ] The results also showed that there existed certain similarities in biological characteristics between the pathogens of silkworm white muscardine from some silkworm rearing areas and from biocontrol agent B. bassiana. Meanwhile, some diversity also existed in B. bassiana strains from the same silkworm rearing area. Therefore, it is implied that the source of white muscardine is rather complex, and further investigation is thus needed to ascertain whether the two biocontrol agents Bb2 and Bb7 belong to B. bassiana.%对广州市森林保护用生防白僵菌Bb2、华南农业大学保存生防用白僵菌Bb7以及收集自广东省化州蚕种场、化州市笪桥村、翁源县硝村等蚕区的3株家蚕白僵菌分离株(Bb8

  20. Potential of Entomopathogen Beauveria bassiana for Controlling Fall Webworm Hyphantria cunea%应用白僵菌防治美国白蛾的潜力

    Institute of Scientific and Technical Information of China (English)

    李会平; 黄秋娴; 王婧; 李雯

    2015-01-01

    [Objective]Hyphantria cunea ( Drury) is a serious destructive insect pest with widespread distribution in China. It usually lives and attacks fruit and ornamental trees,especially broad-leaved trees. The pest has become a big problem to agriculture and forestry in northern China. Up to now,control strategies for H. cunea populations still depend to a large extent on the use of insecticides. However,the resistance development to insecticides and the insecticide-caused environmental pollution stimulate people to search for safely alternative methods to control the pests. Beauveria bassiana, as one of the most common entomopathogenic fungi,has long been recognized as a potential biocontrol agent and actively been developed to control various pest insects. However,there is significant difference in pathogenicity among different strains of B. bassiana,and there exists obvious host specificity in different strains. Thus,it is feasible to enhance the specialization and the pathogenicity of B. bassiana to some kind of pests through artificial orientation training. The virulence of five B. bassiana isolates from different hosts or habitats to larvae of H. cunea was studied to exploit the potentiality of B. bassiana for controlling H. cunea. [Method] 5-instar larvae of H. cunea were used to investigate the effectiveness of five B. bassiana isolates,BS04,BS05 and BS08 from soil,BH01 from larvae of H. cunea,BI05 from larvae of Apriona germari (Hope). The larvae of H. cunea were sprayed with 1 × 108conidia·mL -1conidial suspension of diferent B. bassiana isolates using POTTER spray tower. Mortality was recorded daily for eight days. For the highly virulent isolates identified in the above bioassay,larvae were sprayed with five serial concentrations of conidia (1 × 104, 1 × 105 ,1 × 106 ,1 × 107 and 1 × 108 conidia · mL -1 ) to determine the lethal concentration ( LC50 ) . In the field experiment,there were three treatments,including 8 × 108 conidia·mL -1 of B

  1. Similarity of avian paramyxovirus serotype 1 isolates of low virulence for chickens obtained from contaminated poultry vaccines and from poultry flocks

    DEFF Research Database (Denmark)

    Jørgensen, Poul Henrik; Handberg, K. J.; Ahrens, Peter

    2000-01-01

    flocks. one of which had been vaccinated with two of the contaminated vaccines. The flocks belonged to the same hatchery organisation. A comparison of viral F0 gene sequences and typing of virus isolates with a panel of monoclonal antibodies showed that the vaccine and field isolates were identical....

  2. The characteristics of keratomycosis by Beauveria bassiana and its successful treatment with antimycotic agents

    Directory of Open Access Journals (Sweden)

    Hiroko Sonoyama

    2008-10-01

    Full Text Available Hiroko Sonoyama1, Kaoru Araki-Sasaki1, Shigeyasu Kazama1, Tsutomu Kawasaki1, Hidenao Ideta1, Atsuko Sunada2, Seishi Asari2, Yoshitsugu Inoue3, Kozaburo Hayashi41Ideta Eye Hospital, Kumamoto, Kumamoto, Japan; 2Department of Laboratory for Clinical Investigation, Osaka University Hospital, Suita, Osaka, Japan; 3Division of Ophthalmology and Visual Science, Department of Medicine of Sensory and Motor Organ, Tottori University, Yonago, Tottori, Japan; 4Immunology and Virology Section Lab, Immunology, NEI, NIH, Bethesda, MD, USAAbstract: Clinical findings and treatment of keratomycosis caused by Beauveria bassiana, an entomopathogenic filamentous fungus, are described for an 80-year-old woman, who was referred to the hospital for ocular pain and redness on the 9th day after an ocular injury caused by the frame of her glasses. She had a long history of recurrent diabetic iritis and continuously used topical antibiotics and corticosteroids. At her first visit, a slit-lamp examination indicated a corneal ulcer confined within the superficial stromal layer, along with a slight infiltration and edema. Only a very few inflammatory cells were seen in the anterior chamber. Direct microscopic examination of corneal scrapings revealed septate fungal hyphae with zig-zag rachis and budding that was subsequently identified as B. bassiana by slide culture. Topical voriconazole with miconazole, pimaricin and oral itraconazole were effective and the lesion disappeared leaving only a mild scar at 2 months. The sensitivity of B. bassiana to various antimycotic agents was confirmed by broth microdilution, agar dilution with the Clinical Laboratory Standard Institute standard, and a disk method using topically applied concentrations. B. bassiana, which exhibits a characteristic appearance in smears and causes superficial keratomycosis, is sensitive to voriconazole with miconazole, pimaricin, and itraconazole.Keywords: Beauveria bassiana, keratomycosis, filamentous

  3. 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

  4. Application of the Beauveria bassiana Strain for the Enantioselective Oxidation of the Diethyl 1-Hydroxy-1-Phenylmethanephosphonate

    OpenAIRE

    2010-01-01

    Biotransformation of diethyl 1-hydroxy-1-phenylmethanephosphonate using fungi Beauveria bassiana allowed resolving the racemic mixture of the substrate and due to the biocatalyst and reaction conditions modifications, leading to desired optical isomer.

  5. Joint action of Beauveria bassiana and the insect growth regulators diflubenzuron and novaluron, on the migratory locust, Locusta migratoria

    Science.gov (United States)

    Beauveria bassiana (Balsamo) Vuillemin and sublethal concentrations of the insect growth regulators (IGR) diflubenzuron and novaluron were applied simultaneously and sequentially to second instar Locusta migratoria migratorioides (Sauss.) to determine the interaction between these materials and an e...

  6. 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 ...

  7. 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

  8. Plant extracts on Hypothenemus hampei (Coleoptera: Curculionidae and Beauveria bassianaExtratos vegetais sobre Hypothenemus hampei (Coleoptera: Curculionidae e Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    Janaina Zorzetti

    2012-12-01

    Full Text Available Looking for alternatives to pesticides for Hypothenemus hampei control, the objective of this study was to evaluate the potential of aqueous and ethanolic plant extracts of Moringa oleifera (Moringa and Tephrosia purpurea (tephrosia (seeds, leaves and roots and Melia azedarach (cinnamon, Nerium oleander (oleander and Azadirachta indica (neem (leaves only, on mortality and repellency of H. hampei and its compatibility with Beauveria bassiana, an important natural enemy of this pest. To assess the mortality, coffee leaves (Coffea arabica L were treated by immersion in a solution of endosulfan and plant extracts at a concentration of 10% and then offered to adults of H. hampei. The repellency was evaluated in multiple-choice tests and in no-choice tests among coffee fruit treated and untreated The compatibility between extracts and Beauveria bassiana (CG 452 was analyzed by quantifying germination, colony forming units, growth and yield / productivity of conidia. The highest mortalities were observed when leaves were treated with ethanolic extract of T. purpurea (leaves which did not differ from endosulfan, and aqueous and ethanolic extracts from M. oleifera seeds . In free-choice tests, all the ethanolic extracts showed repellent action, being higher for M. oleifera (root and T. purpurea (seed. The aqueous extracts of M. oleifera (leaves and seeds and N. oleander (leaves showed the highest repellency. In no-choice tests the highest repellency level was for coffee fruits treated with A. indica (leaves. The ethanolic extract of M. oleifera leaves negatively affected B. bassiana germination. These studies showed the potential of these plant extracts for use in the field as an alternative to chemical control, once they are also selective for B. bassiana. Visando alternativas ao uso de agrotóxicos no controle de Hypothenemus hampei, o objetivo deste trabalho foi avaliar o potencial de extratos vegetais aquosos e etanólicos de Moringa oleifera

  9. 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.

  10. An algorithm that predicts the viability and the yield of human hepatocytes isolated from remnant liver pieces obtained from liver resections.

    Science.gov (United States)

    Lee, Serene M L; Schelcher, Celine; Laubender, Rüdiger P; Fröse, Natalja; Thasler, Reinhard M K; Schiergens, Tobias S; Mansmann, Ulrich; Thasler, Wolfgang E

    2014-01-01

    Isolated human primary hepatocytes are an essential in vitro model for basic and clinical research. For successful application as a model, isolated hepatocytes need to have a good viability and be available in sufficient yield. Therefore, this study aims to identify donor characteristics, intra-operative factors, tissue processing and cell isolation parameters that affect the viability and yield of human hepatocytes. Remnant liver pieces from tissue designated as surgical waste were collected from 1034 donors with informed consent. Human hepatocytes were isolated by a two-step collagenase perfusion technique with modifications and hepatocyte yield and viability were subsequently determined. The accompanying patient data was collected and entered into a database. Univariate analyses found that the viability and the yield of hepatocytes were affected by many of the variables examined. Multivariate analyses were then carried out to confirm the factors that have a significant relationship with the viability and the yield. It was found that the viability of hepatocytes was significantly decreased by the presence of fibrosis, liver fat and with increasing gamma-glutamyltranspeptidase activity and bilirubin content. Yield was significantly decreased by the presence of liver fat, septal fibrosis, with increasing aspartate aminotransferase activity, cold ischemia times and weight of perfused liver. However, yield was significantly increased by chemotherapy treatment. In conclusion, this study determined the variables that have a significant effect on the viability and the yield of isolated human hepatocytes. These variables have been used to generate an algorithm that can calculate projected viability and yield of isolated human hepatocytes. In this way, projected viability can be determined even before isolation of hepatocytes, so that donors that result in high viability and yield can be identified. Further, if the viability and yield of the isolated hepatocytes is lower

  11. Evaluation of Genetic Polymorphism of Leishmania (V.) braziliensis Isolates Obtained from the Same Patient before and after Therapeutic Failure or Reactivation of Cutaneous Lesions

    Science.gov (United States)

    Baptista, Cibele; Schubach, Armando de Oliveira; Madeira, Maria de Fatima; de Freitas Campos Miranda, Luciana; Guimarães de Souza Pinto, Andressa; Helena da Silva Barros, Juliana; Conceição-Silva, Fatima; Fernandes Pimentel, Maria Ines; da Silva Pacheco, Raquel

    2012-01-01

    The aim of this study was to investigate genetic polymorphism in Leishmania braziliensis population previously typed through isoenzyme electrophoresis, isolated from the same patient in two different moments: (A) before the beginning of treatment and (B) after treatment failure to meglumine antimoniate or reactivation after successful initial treatment. Fifteen pairs of isolates were assessed using the polymorphic molecular marker LSSP-PCR and following the phenetic analysis. The genetic profiles of the 30 samples were grouped in four clusters. Only two patients presented total identity in the A and B isolates. Most isolates presented similarity coefficients varying from 0.63 to 0.91. In this group of patients genetic polymorphisms could be observed indicating low similarity between the pairs of isolates. The results demonstrate the existence of genetic polymorphism between the samples isolated before treatment and after reactivation or treatment failure, suggesting a possible differentiation of the structure of the original parasite population which could be involved in the mechanisms of resistance to treatment or reactivation of lesions in the ATL. This phenomenon is important, although other factors also could be involved in this context and are discussed in this paper. PMID:23304168

  12. Isolation

    DEFF Research Database (Denmark)

    Agerholm, Frank Juul

    2011-01-01

    Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!......Næringsstoffet har i dette nummer sat fokus på ”velvære i vinterkulden”, ”indendørsaktiviteter” og ”fedtafgift”. I klummen vises det, at disse tre fokusområder, der for en umiddelbar betragtning måske nok synes noget uensartede, falder sammen i ét tema: Isolation!...

  13. Bioassay and Scanning Electron Microscopic Observations Reveal High Virulence of Entomopathogenic Fungus, Beauveria bassiana, on the Onion Maggot (Diptera: Anthomyiidae) Adults.

    Science.gov (United States)

    Zhang, Hui; Wu, Shengyong; Xing, Zhenlong; Wang, Xiaoqing; Lei, Zhongren

    2016-12-01

    When flies were dipped in 1 × 10(8) conidia/ml conidia suspensions and then kept in the incubator (22 ± 1 °C, 70 ± 5% RH), scanning electron microscope observations revealed that, at 2 h, the majority of adhering Beauveria bassiana conidia were attached to either the wing surface or the interstitial area between the macrochaetae on the thorax and abdomen of the onion maggot adults. Germ tubes were being produced and had oriented toward the cuticle by 18 h. Penetration of the insect cuticle had occurred by 36 h, and by 48 h, germ tubes had completely penetrated the cuticle. Fungal mycelia had emerged from the insect body and were proliferating after 72 h. The superficial area and structure of the wings and macrochaetae may facilitate the attachment of conidia and enable effective penetration. The susceptibility of adults to 12 isolates, at a concentration of 1 × 10(7) conidia/ml, was tested in laboratory experiments. Eight of the more potent strains caused in excess of 85% adult mortality 8 d post inoculation, while the median lethal time (LT50) of these strains was <6 d. The virulence of the more effective strains was further tested, and the median lethal concentrations (LC50) were calculated by exposing adults to doses ranging from 10(3)-10(7) conidia/ml. The lowest LC50 value, found in the isolate XJWLMQ-32, for the adults was 3.87 × 10(3) conidia/ml. These results demonstrate that some B. bassiana strains are highly virulent to onion maggot adults and should be considered as potential biocontrol agents against the adult flies.

  14. Vegetative compatibility groups and sexual reproduction among Spanish Monilinia fructicola isolates obtained from peach and nectarine orchards, but not Monilinia laxa.

    Science.gov (United States)

    De Cal, Antonieta; Egüen, Beatriz; Melgarejo, Paloma

    2014-01-01

    The frequency of occurrence of Monilinia fructicola in the Ebro Valley, Spain has increased since its first appearance in 2006, and M. fructicola has displaced Monilinia laxa, the native species which is the main cause of brown rot in peaches in this valley. In order to determine the characteristics that may be related to the displacement, we studied the capacity to generate new genotypic combinations of M. fructicola under laboratory conditions. The morphology and parasitic ability from ten field isolates of M. fructicola and M. laxa collected from three different orchards in the valley, and sampling from five different lesions were studied. Nitrate-non-utilising (nit) mutants were generated in order to test the isolates for vegetative compatibility which was done by assessing their colony growth when cultured singly or in pairs on media that contained different nitrogen sources. For the M. fructicola isolates, five vegetative compatibility groups (VCGs) were identified using the nit mutants and six VCGs were identified when they were grown on potato dextrose agar dishes. In all instances, the vegetatively compatible M. fructicola isolates came mainly from the same orchard. Only one VCG displays the same morphological and competition characteristics. No VCGs were identified among the M. laxa isolates. We did not find any apothecia of M. laxa and M. fructicola isolates in the soil of the three orchards, but we were able to produce apothecia of M. fructicola in the laboratory. Our finding of sexual reproduction and VCGs in the M. fructicola isolates suggests that the genetic variability of M. fructicola could be maintained by sexual and/or parasexual recombination.

  15. Estudio de las condiciones de mezclado en fermentador para la producción de blastosporas de Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    Cipriano García Gutiérrez

    2013-12-01

    had the best values with a ratio of diameter of the impeller/vessel (DI/DT of 0.43, KLa = 9.5-208 h-1 and tM = 1.0-3.0 s, as a result, it was selected for the production of blastospores of Beauveria bassiana, using molasses as carbon source. We studied the mixing conditions, using different combinations of impellers, of type Rushton, Lightnin and Maxflo under a factorial experimental design 32. The propagation time was of 4 days, working volume was 4l, 10% inoculum (1x106 blastospores/ml, temperature of 30°C, agitation of 400-500 rpm, aeration 0.5-1.0 vvm, and pH of 5.4. The fungus growth was better using a combination of impellers Rushton-Maxflo at 400 rpm and 1.0 vvm (F = 10.324, p £ 0.0123 (LSD = 0.585 obtaining a concentration of 1.2x109blastospores/ml, 2.2 g/l biomass and 2.48 g/l of substrate consumption (Y x/s = 0.89. The mixing conditions and the parameters obtained can be applied to optimize the blastospore  productions of B. bassiana to fermenter level in the experimental production of bioinsecticides.Key words: liquid fermentation, aeration, agitation, entomopathogenic fungus.

  16. Modulation of conidia production and expression of the gene bbrgs1 from Beauveria bassiana by oxygen pulses and light.

    Science.gov (United States)

    Rodriguez-Gomez, Divanery; Marcial-Quino, Jaime; Loera, Octavio

    2015-09-01

    Light and oxidant states affect the conidiation in diverse fungi, although the response has not been described when both stimuli are applied simultaneously. Conidial production and quality in Beauveria bassiana were analysed under four conditions for a wild-type (wt) strain and a previously isolated mutant (mt): normal atmosphere (21% O2; NA) or oxygen-enriched pulses (26% O2; OEP), with either light (L) or darkness (D). The response was complemented by following the expression of the bbrgs1 gene, encoding a regulator of the G-protein signal associated to conidia production. Conidiation was not significantly affected in the mutant strain by any condition (highest value with NA-L: 2.7×10(8)concm(-2)). Relative to maximal levels under NA (NA-D: 4×10(7)concm(2)), the wt strain diminished conidiation by 34-fold under OEP. The expression of bbrgs1 was higher (up to 188 times) in the mutant strain in every condition relative to the wt strain, in fact expression levels were consistent with the conidiation yields between strains. Viability and hydrophobicity were less affected by culture conditions, although pathogenicity parameters improved in conidia from OEP. The response to OEP, either with light or darkness, was strain-dependent for conidial production, viability, hydrophobicity and infectivity of conidia, then these parameters could be modulated in mass production processes. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Spore UV and acceleration resistance of endolithic Bacillus pumilus and Bacillus subtilis isolates obtained from Sonoran desert basalt: implications for lithopanspermia.

    Science.gov (United States)

    Benardini, James N; Sawyer, John; Venkateswaran, Kasthuri; Nicholson, Wayne L

    2003-01-01

    Bacterial spores have been used as model systems for studying the theory of interplanetary transport of life by natural processes such as asteroidal or cometary impacts (i.e., lithopanspermia). Because current spallation theory predicts that near-surface rocks are ideal candidates for planetary ejection and surface basalts are widely distributed throughout the rocky planets, we isolated spore-forming bacteria from the interior of near-subsurface basalt rocks collected in the Sonoran desert near Tucson, Arizona. Spores were found to inhabit basalt at very low concentrations (basalt samples. Populations of purified spores prepared from the isolated strains were subjected to 254-nm UV and ballistics tests in order to assess their resistance to UV radiation and to extreme acceleration shock, two proposed lethal factors for spores during interplanetary transfer. Specific natural isolates of B. pumilus were found to be substantially more resistant to UV and extreme acceleration than were reference laboratory strains of B. subtilis, the benchmark organism, suggesting that spores of environmental B. pumilus isolates may be more likely to survive the rigors of interplanetary transfer.

  18. The effect of prolonged storage on the virulence of isolates of Bacillus anthracis obtained from environmental and animal sources in the Kars Region of Turkey.

    Science.gov (United States)

    Buyuk, Fatih; Sahin, Mitat; Cooper, Callum; Celebi, Ozgur; Saglam, Aliye Gulmez; Baillie, Les; Celik, Elif; Akca, Dogan; Otlu, Salih

    2015-07-01

    The stability of the plasmid-mediated virulence factors of Bacillus anthracis, a tripartite toxin located on pXO1 and an antiphagocytic capsule encoded by genes located on pXO2, following long-term storage was investigated. A collection of 159 isolates of B. anthracis were collected from the Kars region of Turkey between 2000 and 2013 and stored at -20°C in Brucella broth supplemented with 20% glycerine. A total of 142 isolates were recovered of which one failed to express a capsule upon primary culture. A further 35 isolates yielded a mixture of mucoid and non-mucoid colonies; the majority of which had lost the pXO2 plasmid as determined by PCR analysis. Results would suggest that pXO2 is more unstable than pXO1 and that this instability increases with the length of storage. It is possible that the pXO2-deficient isolates of B. anthracis described here could be developed into a vaccine to treat at risk animals in the Kars region as many animal vaccines are based upon pXO2 deficiency.

  19. A user friendly method to isolate and single spore the fungi Magnaporthe oryzae and Magnaporthe grisea obtained from diseased field samples

    Science.gov (United States)

    The fungus Magnaporthe oryzae is the causal agent for a wide range of plant diseases including diseases of rice, wheat, rye grass, turfgrass and pearl millet. A simple robust procedure for fungal isolation is not publicly available. In the present study, a user friendly method was developed to iso...

  20. [In-vitro susceptibilites to levofloxacin and various antibacterial agents of 18,639 clinical isolates obtained from 77 centers in 2004].

    Science.gov (United States)

    Yamaguchi, Keizo; Ohno, Akira; Ishii, Yoshikazu; Tateda, Kazuhiro; Iwata, Morihiro; Kanda, Makoto; Tsujio, Yoshiko; Kimoto, Hiroya; Kaimori, Mitsuomi; Nakamura, Toshihiko; Kawamura, Chizuko; Nishimura, Masaharu; Akizawa, Koji; Katayama, Yosei; Matsuda, Keiko; Hayashi, Tasuku; Yasujima, Minoru; Kasai, Takeshi; Kimura, Masahiko; Tominaga, Makoto; Miki, Makoto; Nakanowatari, Susumu; Nakagawa, Takuo; Kaku, Mitsuo; Kanemitsu, Keiji; Kunishima, Hiroyuki; Toyoshima, Shunkoh; Sakurai, Masanori; Shiotani, Joji; Sugita, Akihiro; Ito, Tatsumi; Okada, Jun; Suwabe, Akira; Yamahata, Kumiko; Yoneyama, Akiko; Kumasaka, Kazunari; Yamane, Nobuo; Koike, Kazuhiko; Ieiri, Tamio; Kominami, Hidenori; Yamada, Toshiyuki; Oguri, Toyoko; Itoh, Kouichi; Watanabe, Kiyoaki; Kobayashi, Yoshio; Ohtake, Teruko; Uchida, Takashi; Totsuka, Kyoichi; Murakami, Masami; Yomoda, Sachie; Takahashi, Ayako; Okamoto, Hideyuki; Inuzuka, Kazuhisa; Yamazaki, Kenichiro; Gonda, Hideo; Yamashita, Takanori; Yamaguchi, Ikuo; Okada, Motoi; Ikari, Hiromi; Kurosawa, Naomi; Fujimoto, Yoshinori; Ishigo, Shiomi; Asano, Yuko; Mikio, Mori; Kano, Ichino; Nagano, Eiko; Kageyama, Fumio; Shaku, Etsuko; Kanno, Harushige; Aihara, Masanori; Gemma, Hitoshi; Uemura, Keiichi; Miyajima, Eiji; Maesaki, Shighefumi; Hashikita, Giichi; Horii, Toshinobu; Sumitomo, Midori; Yoshimura, Hitoshi; Hiraoka, Minoru; Wada, Hideo; Yuzuki, Yosuke; Ikeda, Norio; Baba, Hisashi; Soma, Masayuki; Yamamoto, Tsuyoshi; Ichiyama, Satoshi; Kinosita, Syohiro; Kawano, Seiji; Fujita, Shinichi; Kageoka, Takeshi; Hongo, Toshiharu; Okabe, Hidetoshi; Tatewaki, Kenichi; Moro, Kunihiko; Oka, Mikio; Niki, Yoshihito; Yoshida, Haruyoshi; Yamashita, Masanobu; Kusano, Nobuchika; Mihara, Eiichiro; Nose, Motoko; Fushiwaki, Takeshi; Kuwabara, Masao; Fujiue, Yoshihiro; Shimuzu, Akira; Takubo, Takayuki; Kusakabe, Tadashi; Hinoda, Yuji; Tanaka, Nobuaki; Takahashi, Hakuo; Heijyou, Hitoshi; Okazaki, Toshiro; Asai, Koji; Kawahara, Kunimitsu; Masuda, Junichi; Sano, Reiko; Taminato, Tomohiko; Negayama, Kiyoshi; Matsuo, Syuji; Komatsu, Masaru; Sugiura, Tetsuro; Murase, Mitsuharu; Hiramatsu, Kazufumi; Yamane, Nobuhisa; Nakasone, Isao; Hirakata, Yoichi; Kohno, Shigeru; Aizawa, Hisamichi; Honda, Junichi; Hamazaki, Naotaka; Okayama, Akihiko; Ono, Junko; Aoki, Yosuke; Okada, Kaoru; Miyanohara, Hiroaki

    2006-12-01

    A total of 18,639 clinical isolates in 19 species collected from 77 centers during 2004 in Japan were tested for their susceptibility to fluoroquinolones (FQs) and other selected antibiotics. The common respiratory pathogens, Streptococcus pyogenes, Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae showed a high susceptible rate against FQs. The isolation rate of beta lactamase non-producing ampicillin-resistant H. influenzae was approximately three times as large as those of western countries. Most strains of Enterobacteriaceae were also susceptible to FQs. The resistance rate of Escherichia coli against FQs has however been rapidly increasing so far as we surveyed since 1994. The FQs-resistant rate in methicillin-resistant Staphylococcus aureus (MRSA) showed approximately 90% except for 36%. of sitafloxacin while FQs-resistant rate in methicillin-susceptible S. aureus (MSSA) was around 5%. The FQs-resistant rate of methicillin-resistant coagulase negative Staphylococci (MRCNS) was also higher than that of methicillin-susceptible coagulase negative Staphylococci (MSCNS), however, it was lower than that of MRSA. In Pseudomonas aeruginosa clinical isolates, 32-34% from UTI and 15-19% of from RTI was resistant to FQs. Acinetobacter spp. showed a high susceptibility to FQs. Although FQs-resistant Neisseria gonorrhoeae have not been increased in western countries, it is remarkably high in Japan. In this survey, isolates of approximately 85% was resistant to FQs.

  1. [Spa types and antibiotic resistance of Staphylococcus aureus bloodstream isolates obtained form patients of the University Clinical Center in Gdańsk].

    Science.gov (United States)

    Wiśniewska, Katarzyna; Kasprzyk, Joanna; Piechowicz, Lidia; Bronk, Marek; Swieć, Krystyna

    2013-01-01

    Staphylococcus aureus is a leading cause of bloodstream infections. For epidemiological investigations of this bacteria spa genotyping is used as the method which has a high discriminatory power and gives results that can be easily compared between laboratories. In contrast to methicillin-resistant S.aureus (MRSA), relatively little is known about spa types among methicillin-susceptible strains (MSSA). We used spa typing and antibiotic resistance patterns analysis for retrospective study of S.aureus bloodstream isolates population from the University Clinical Centre (UCC) in Gdańsk. The study was performed on 53 isolates from patients of 19 different units/ departments of the UCC. The isolates were tested for the susceptibility to antimicrobial agents. Spa typing was performed on the basis of the sequence analysis of the polymorphic X region of the protein A gene (spa) amplified form the isolates. Spa types were determined by Ridom Staph Type software and were clustered into spa-CCs (clonal complexes) using the algorithm BURP-based upon repeat pattern. MLST (Multilocus Sequence Typing) clonal complexes were predicted from BURP analysis by the Ridom SpaServer database. In MRSA the staphylococcal chromosomal casette (SCC) mec was determined, Spa-typing yielded 26 types. Six spa-CC and seven singletons were identified. The most frequent was spa-CC021involving 38% of isolates. The CC021 consisted of 7 spa types and the most common was t021 corresponding with MLST-CC30. The second frequent was singleton, related to MLST-CC1, with only one type t127. There were 3 MRSA isolates in the population. The MRSA strains were identified as different spa types: t003/ SCCmecII, t008/SCCmecIV and clonally related to MSSA t032/SCCmecIV. No one MRSA strains belonged to spa-CC021. The spa clonal cluster corresponding with widely distributed among invasive S.aureus strains in Europe MLST-CC30 was found as the most frequent among S.aureus bloodstream isolates from the UCC. Occurrence of

  2. Sensitivity ofBeauveria bassiana to solanine and tomatine : Plant defensive chemicals inhibit an insect pathogen.

    Science.gov (United States)

    Costa, S D; Gaugler, R R

    1989-02-01

    The alkaloids solanine and tomatine and the polyene antibiotic nystatin were tested in vitro against the entomopathogenic fungusBeauveria bassiana. Nystatin was the most inhibitory compound tested, reducing colony formation, growth, and development of conidiaphores to a greater degree and at lower concentrations than the alkaloids. Tomatine inhibited colony formation and growth more than solanine, which had relatively little effect on the fungus. The toxicity of tomatine suggests that germination of conidia and subsequent hyphal growth would be inhibited when an insect consumes conidia along with foliage containing 0.100 mg/g (fresh weight) of this compound. The sensitivity ofB. bassiana to alkaloids appears to be in the middle of the range found with other fungi.

  3. 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.

  4. Fungal Keratitis Due to Beauveria bassiana in a Contact Lenses Wearer and Review of Published Reports.

    Science.gov (United States)

    Lara Oya, Ana; Medialdea Hurtado, María Eloisa; Rojo Martín, María Dolores; Aguilera Pérez, Antonia; Alastruey-Izquierdo, Ana; Miranda Casas, Consuelo; Rubio Prats, Marina; Medialdea Marcos, Santiago; Navarro Marí, José María

    2016-10-01

    Fungal keratitis is a severe ocular infection that primarily affects subjects engaged in outdoor activities. Risk factors include allergic conjunctivitis, previous eye surgery, previous treatment with wide-spectrum antimicrobial agents and corticosteroids and using contact lenses. Corneal infection is usually secondary to trauma involving organic material, which is often the only predisposing factor. Early diagnosis based on clinical examination and microbiological investigation (microscopy, cultures and molecular techniques) is crucial to selecting the appropriate antifungal therapy and prevent progression. We report the case of a patient with keratitis due to Beauveria bassiana, an opportunistic and entomopathogenic filamentous fungus that is used as a biological insecticide and which is a rare cause of corneal infection. We review previous cases reports of B. bassiana keratitis published and its main features to compare with our case, a female occasional agriculture worker who had not suffered any trauma involving organic material. The patient received topical and oral antifungal therapy and debridement surgery, with a satisfactory outcome.

  5. Production of antibacterial Bombyx mori cecropin A in mealworm-pathogenic Beauveria bassiana ERL1170.

    Science.gov (United States)

    Lee, Se Jin; Yu, Jeong Seon; Parker, Bruce L; Skinner, Margaret; Je, Yeon Ho; Kim, Jae Su

    2015-01-01

    Efforts are underway to produce antimicrobial peptides in yellow mealworms (Tenebrio molitor), which can be developed as more effective and safer animal feed additives. In this work, we expressed Bombyx mori (Bm) cecropin-A in mealworms by the infection of transformed entomopathogenic Beauveria bassiana ERL1170. The active domain of Bm cecropin A gene was tagged with a signal sequence of B. bassiana for extracellular secretion, and the fragment was inserted into ERL1170 by the restriction enzyme-mediated integration method. Transformant D-6 showed antibacterial activity against Bacillus subtilis and Listeria monocytogenes. Against T. molitor larvae, D-6 had similar mortality to wild-type, and D6-infected mealworm suspension showed strong antibacterial activity against the two bacteria, but not in the wild-type-infected mealworms, thereby increasing the value of mealworms as animal feed additives.

  6. Molecular typing of isolates obtained from aborted foetuses in Brucella-free Holstein dairy cattle herd after immunisation with Brucella abortus RB51 vaccine in Egypt.

    Science.gov (United States)

    Wareth, Gamal; Melzer, Falk; Böttcher, Denny; El-Diasty, Mohamed; El-Beskawy, Mohamed; Rasheed, Nesma; Schmoock, Gernot; Roesler, Uwe; Sprague, Lisa D; Neubauer, Heinrich

    2016-12-01

    Bovine brucellosis is endemic in Egypt in spite of application of surveillance and control measures. An increase of abortions was reported in a Holstein dairy cattle herd with 600 animals in Damietta governorate in Egypt after immunisation with Brucella (B.) abortus RB51 vaccine. Twenty one (10.6%) of 197 vaccinated cows aborted after 3 months. All aborted cows had been tested seronegative for brucellosis in the past 3 years. B. abortus was isolated from four foetuses. Conventional biochemical and bacteriological identification and polymerase chain reaction (PCR) confirmed two B. abortus biovar (bv.) 1 smooth and two B. abortus rough strains. None of the B. abortus isolates were identified as RB51. Genotyping analysis by multiple locus of variable number tandem repeats analysis based on 16 markers (MLVA-16) revealed two different profiles with low genetic diversity. B. abortus bv1 was introduced in the herd and caused abortions.

  7. 白僵菌RAPD-PCR反应体系优化%RAPD-PCR Reaction System for Beauveria bassiana

    Institute of Scientific and Technical Information of China (English)

    魏晓鹏; 李会平; 黄大庄; 唐秀光

    2012-01-01

    为了建立一套适宜于白僵菌(Beauveria bassiana)退化研究的RAPD-PCR反应体系及反应程序,通过采用L16( 45)正交试验及退火温度和循环次数的单因素优化对反应体系中的各因素进行优化组合.结果表明:20μL PCR反应体系及反应程序中各因素优化组合为,10×Buffer 2 μL,MgCl2(25 mmol/L)2.4 μL,4种dNTP(各2.5mmol/L)0.8 μL,随机引物(10 μmol/L)1.4 μL,TaqDNA聚合酶(5 U/μL)0.4 μL,模板DNA( 10 mg/L)1 μL.反应条件为,94℃预变性2 min,94℃变性30 s,38℃退火40 s,72℃延伸1 min,循环次数40次,72℃延伸5 min.%An experiment was conducted to establish an optimal PCR (polymerase chain reaction) reaction system and procedure for the degradation of Beauveria bassiana. Single factor test and L16(45) orthogonal experiment were used to optimize the combination of factors for the reaction system. The optimum factor combination was obtained with 20 μL reaction volume containing 2 μL l0×Buffer, 2.4 μL MgCl2(25 mmol/L) , 0. 8 μL four types of dNTPs (each 2. 5 mmol/L) , 1.4μL random primer (10 μmol/L) , 0.4 μL Taq polymerase (5 U/ μL) , and 1 μL template DNA (10 mg/L). Reaction conditions were as follows; predenaturing at 94 degrees C for 2 min, followed by 40 cycles of denaturing at 94 degrees C for 30 s, annealing at 38 degrees C for 40 s, extension at 72 degrees C for 1 min, and final extension at 72 degrees C for 5 min.

  8. 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)

  9. Protection of metal artefacts with the formation of metal-oxalates complexes by Beauveria bassiana.

    Directory of Open Access Journals (Sweden)

    Edith eJoseph

    2012-01-01

    Full Text Available Several fungi present high tolerance to toxic metals and some are able to transform metals into metal-oxalate complexes. In this study, the ability of Beauveria bassiana to produce copper oxalates was evaluated in vitro. Growth performance was tested on various copper-containing media. B. bassiana proved highly resistant to copper, tolerating concentrations of up to 20 g.L-1, and precipitating copper oxalates on all media tested. Chromatographic analyses showed that this species produced oxalic acid as sole metal chelator. The production of metal-oxalates can be used in the restoration and conservation of archaeological and modern metal artefacts. The production of copper-oxalates was confirmed directly using metallic pieces (both archaeological and modern. The conversion of corrosion products into copper oxalates was demonstrated as well. In order to assess whether the capability of B. bassiana to produce metal-oxalates could be applied to other metals, iron and silver were tested as well. Iron appears to be directly sequestered in the wall of the fungal hyphae forming oxalates and probably goethite. However, the formation of a homogeneous layer on the object is not yet optimal. Silver nitrate was extracellularly reduced into nanoparticles of elemental silver by an unknown mechanism. The production of copper oxalates is immediately applicable for the conservation of copper-based artefacts. For iron and silver this is not yet the case. However, the vast ability of B. bassiana to transform toxic metals using different immobilization mechanisms seems to offer considerable possibilities for industrial applications, such as the bioremediation of contaminated soils or the green synthesis of chemicals.

  10. The mosquito melanization response is implicated in defense against the entomopathogenic fungus Beauveria bassiana.

    Directory of Open Access Journals (Sweden)

    Hassan Yassine

    Full Text Available Mosquito immunity studies have focused mainly on characterizing immune effector mechanisms elicited against parasites, bacteria and more recently, viruses. However, those elicited against entomopathogenic fungi remain poorly understood, despite the ubiquitous nature of these microorganisms and their unique invasion route that bypasses the midgut epithelium, an important immune tissue and physical barrier. Here, we used the malaria vector Anopheles gambiae as a model to investigate the role of melanization, a potent immune effector mechanism of arthropods, in mosquito defense against the entomopathogenic fungus Beauveria bassiana, using in vivo functional genetic analysis and confocal microscopy. The temporal monitoring of fungal growth in mosquitoes injected with B. bassiana conidia showed that melanin eventually formed on all stages, including conidia, germ tubes and hyphae, except the single cell hyphal bodies. Nevertheless, melanin rarely aborted the growth of any of these stages and the mycelium continued growing despite being melanized. Silencing TEP1 and CLIPA8, key positive regulators of Plasmodium and bacterial melanization in A. gambiae, abolished completely melanin formation on hyphae but not on germinating conidia or germ tubes. The detection of a layer of hemocytes surrounding germinating conidia but not hyphae suggested that melanization of early fungal stages is cell-mediated while that of late stages is a humoral response dependent on TEP1 and CLIPA8. Microscopic analysis revealed specific association of TEP1 with surfaces of hyphae and the requirement of both, TEP1 and CLIPA8, for recruiting phenoloxidase to these surfaces. Finally, fungal proliferation was more rapid in TEP1 and CLIPA8 knockdown mosquitoes which exhibited increased sensitivity to natural B. bassiana infections than controls. In sum, the mosquito melanization response retards significantly B. bassiana growth and dissemination, a finding that may be exploited to

  11. The mosquito melanization response is implicated in defense against the entomopathogenic fungus Beauveria bassiana.

    Science.gov (United States)

    Yassine, Hassan; Kamareddine, Layla; Osta, Mike A

    2012-01-01

    Mosquito immunity studies have focused mainly on characterizing immune effector mechanisms elicited against parasites, bacteria and more recently, viruses. However, those elicited against entomopathogenic fungi remain poorly understood, despite the ubiquitous nature of these microorganisms and their unique invasion route that bypasses the midgut epithelium, an important immune tissue and physical barrier. Here, we used the malaria vector Anopheles gambiae as a model to investigate the role of melanization, a potent immune effector mechanism of arthropods, in mosquito defense against the entomopathogenic fungus Beauveria bassiana, using in vivo functional genetic analysis and confocal microscopy. The temporal monitoring of fungal growth in mosquitoes injected with B. bassiana conidia showed that melanin eventually formed on all stages, including conidia, germ tubes and hyphae, except the single cell hyphal bodies. Nevertheless, melanin rarely aborted the growth of any of these stages and the mycelium continued growing despite being melanized. Silencing TEP1 and CLIPA8, key positive regulators of Plasmodium and bacterial melanization in A. gambiae, abolished completely melanin formation on hyphae but not on germinating conidia or germ tubes. The detection of a layer of hemocytes surrounding germinating conidia but not hyphae suggested that melanization of early fungal stages is cell-mediated while that of late stages is a humoral response dependent on TEP1 and CLIPA8. Microscopic analysis revealed specific association of TEP1 with surfaces of hyphae and the requirement of both, TEP1 and CLIPA8, for recruiting phenoloxidase to these surfaces. Finally, fungal proliferation was more rapid in TEP1 and CLIPA8 knockdown mosquitoes which exhibited increased sensitivity to natural B. bassiana infections than controls. In sum, the mosquito melanization response retards significantly B. bassiana growth and dissemination, a finding that may be exploited to design transgenic

  12. Soil application of Beauveria bassiana to control Ceratitis capitata in semi field conditions.

    Science.gov (United States)

    Ali, Ali; Sermann, Helga; Lerche, Sandra; Büttner, Carmen

    2009-01-01

    The Mediterranean fruit fly Ceratitis capitata (Wiedemann) is a highly polyphagous pest of economic importance cultures in Syria, as in many other parts of the world. The potential of the entomopathogenic fungus Beauveria bassiona BALS (VUIL.) strain 412 against adults of Mediterranean fruit fly C. capitata was evaluated in semi field conditions during the summer. Soil (5-7 cm high) was filled into plastic container (27 cm x 32 cm). In one container 75 pupae, two days before emergency, were spread uniformly on the soil. Then the pupae were covered with soil (4-5 cm layer). After that, 30 ml suspension of fungal spores (4 x 10(8) spores/ml) was applied to the soil surface using a dash bottle. This corresponded to a spore density of 1.3 x 10(7) spores/cm2 on soil. Water and food (1:4 yeast, sucrose) were placed in the cages for the emerged flies. The semi-field evaluation of B. bassiana revealed a fly mortality of about 46% compared to 16% in the control. In addition 72% of dead flies were moulded in the treatment. These results indicated that the entomopathogenic fungus B. bassiana was pathogen against the adults of C. capitata not only in the laboratory condition but also under field condition. That means B. bassiana could decrease the offspring of C. capitata. Therefore B. bassiana could be an effective factor to control C. capitata in combination with other control methods, used in IPM program in the field.

  13. Bombyx mori cecropin A has a high antifungal activity to entomopathogenic fungus Beauveria bassiana.

    Science.gov (United States)

    Lu, Dingding; Geng, Tao; Hou, Chengxiang; Huang, Yuxia; Qin, Guangxing; Guo, Xijie

    2016-05-25

    A cDNA encoding cecropin A (CecA) was cloned from the larvae of silkworm, Bombyx mori, using RT-PCR. It encodes a protein of 63 amino acids, containing a 22 amino acid signal peptide and a 37 amino acid mat peptide of functional domain. The CecA secondary structure contains two typical amphiphilic α-helices. Real-time qPCR analysis revealed that CecA was expressed in all the tissues tested, including cuticle, fat body, hemocytes, Malpighian tubule, midgut and silk gland in the silkworm larvae with the highest expression in the fat body and hemocytes. The gene expression of B. mori CecA was rapidly induced by Beauveria bassiana challenge and reached maximum levels at 36h after inoculation in third instar larvae. In the fifth instar larvae infected with B. bassiana, the relative expression level of CecA was upregulated in fat body and hemocytes, but not in cuticle, Malpighian tubule, midgut and silk gland. The cDNA segment of the CecA was inserted into the expression plasmid pET-30a(+) to construct a recombinant expression plasmid. Western blot results revealed that his-tagged fusion protein was successfully expressed and purified. Then the mat peptide of CecA was chemically synthesized with C-terminus amidation for in vivo antifungal assay and purity achieved 93.7%. Mass spectrometry and SDS-PAGE showed its molecular weight to be 4046.95Da. Antifungal assays indicated that the B. mori CecA had a high antifungal activity to entomopathogenic fungus B. bassiana both in vitro and in vivo in the silkworm larvae. This is the first report that the CecA is effective to inhibit B. bassiana inside the body of silkworm.

  14. Fitness consequences of larval exposure to Beauveria bassiana on adults of the malaria vector Anopheles stephensi.

    Science.gov (United States)

    Vogels, Chantal B F; Bukhari, Tullu; Koenraadt, Constantianus J M

    2014-06-01

    Entomopathogenic fungi have shown to be effective in biological control of both larval and adult stages of malaria mosquitoes. However, a small fraction of mosquitoes is still able to emerge after treatment with fungus during the larval stage. It remains unclear whether fitness of these adults is affected by the treatment during the larval stage and whether they are still susceptible for another treatment during the adult stage. Therefore, we tested the effects of larval exposure to the entomopathogenic fungus Beauveria bassiana on fitness of surviving Anopheles stephensi females. Furthermore, we tested whether larval exposed females were still susceptible to re-exposure to the fungus during the adult stage. Sex ratio, survival and reproductive success were compared between non-exposed and larval exposed A. stephensi. Comparisons were also made between survival of non-exposed and larval exposed females that were re-exposed to B. bassiana during the adult stage. Larval treatment did not affect sex ratio of emerging mosquitoes. Larval exposed females that were infected died significantly faster and laid equal numbers of eggs from which equal numbers of larvae hatched, compared to non-exposed females. Larval exposed females that were uninfected had equal survival, but laid a significantly larger number of eggs from which a significantly higher number of larvae hatched, compared to non-exposed females. Larval exposed females which were re-exposed to B. bassiana during the adult stage had equal survival as females exposed only during the adult stage. Our results suggest that individual consequences for fitness of larval exposed females depended on whether a fungal infection was acquired during the larval stage. Larval exposed females remained susceptible to re-exposure with B. bassiana during the adult stage, indicating that larval and adult control of malaria mosquitoes with EF are compatible.

  15. Cuticle-degrading proteases and toxins as virulence markers of Beauveria bassiana (Balsamo) Vuillemin.

    Science.gov (United States)

    Cito, Annarita; Barzanti, Gian Paolo; Strangi, Agostino; Francardi, Valeria; Zanfini, Assunta; Dreassi, Elena

    2016-09-01

    Beauveria bassiana is one of the most known entomopathogenic fungal species and its entomopathogenic mechanism involves several bioactive metabolites, mainly cuticle-degrading enzymes and toxic molecules, which are predicted to play a key role as virulence factors. In this study six Beauveria bassiana strains (B 13/I03, B 13/I11, B 13/I49, B 13/I57, B 13/I63, and B 13/I64) were assayed against Tenebrio molitor larvae. Enzymatic activity of total proteases and specifically Pr 1 and Pr 2, as well as the production of toxic compounds were investigated in each fungal strain. Toxins were detected both in vitro-in medium filtrates and mycelia-and in vivo-in Tenebrio molitor larvae infected by the fungal strains tested. B 13/I11 and B 13/I63 strains showed the most significant entomopathogenic activity against Tenebrio molitor larvae (cumulative mortality rate 100 and 97%, respectively; average survival time 5.85 and 6.74 days, respectively). A widely variable and fungal strain-dependent enzymatic activity of total proteases, Pr 1 and Pr 2 was found. Beauvericin, beauvericin A and bassianolide resulted the most prevalent toxins detected in the substrates analyzed. It has been found that an increase of beauvericin content in vivo resulted significantly correlated to a decrease of Tenebrio molitor larvae average survival time in entomopathogenic bioassay (inverse correlation). The involvement of beauvericin in B. bassiana entomopathogenic process is confirmed; in vitro analysis of cuticle degrading proteases activity and toxins production in relation to the methods adopted resulted insufficient for a rapid screening to determine the virulence of B. bassiana strains against Tenebrio molitor larvae.

  16. Laboratory and semi-field evaluation of Beauveria bassiana (Ascomycota: Hypocreales) against the lettuce aphid, Nasonovia ribisnigri (Hemiptera: Aphididae)

    DEFF Research Database (Denmark)

    Shrestha, Govinda; Enkegaard, Annie; Steenberg, Tove

    2015-01-01

    , with the highest effect seen when the alatoid fourth instar of N. ribisnigri was inoculated with B. bassiana. The persistence of B. bassiana conidia on lettuce foliage was not influenced by leaf position. Within 5 days, the cumulative percentage decline in the conidial population was 38% which declined further...

  17. 75 FR 4817 - Beauveria Bassiana Strain GHA; Notice of Receipt of a Request for an Amendment to Delete a Use in...

    Science.gov (United States)

    2010-01-29

    ... From the Federal Register Online via the Government Publishing Office ENVIRONMENTAL PROTECTION AGENCY Beauveria Bassiana Strain GHA; Notice of Receipt of a Request for an Amendment to Delete a Use in... Delete from Label 82074-1 Mycotrol ES Beauveria bassiana Tomato Strain GHA Users of this product...

  18. Highly virulent Beauveria bassiana strains against the two-spotted spider mite, Tetranychus urticae, show no pathogenicity against five phytoseiid mite species.

    Science.gov (United States)

    Wu, Shengyong; Xie, Haicui; Li, Maoye; Xu, Xuenong; Lei, Zhongren

    2016-12-01

    Entomopathogenic fungi and predatory mites can independently contribute to suppressing the two-spotted spider mite, Tetranychus urticae Koch. It is important to assess the risk of possible fungal infections in predators when a combination of them are being considered as a tandem control strategy for suppressing T. urticae. The first part of this study tested 12 Beauveria bassiana isolates for virulence in T. urticae. Strains SCWJ-2, SDDZ-9, LNSZ-26, GZGY-1-3 and WLMQ-32 were found to be the most potent, causing 37.6-49.5% adult corrected mortality at a concentration of 1 × 10(7) m/L conidia 4 days post-treatment. The second part evaluated the pathogenicity of these five strains in five species of predatory phytoseiid mites. The bioassay results indicated that all adult predatory mite mortalities ranged from 7.5 to 9.1% 4 days post-treatment. No viable fungal hyphae were found on predator cadavers. Observations with scanning electron microscopy revealed that conidia were attached to the cuticle of predatory mites within 2-12 h after spraying with strain LNSZ-26, and had germinated within 24-36 h. After 48 h, conidia had gradually been shed from the mites, after none of the conidia had penetrated the cuticular surfaces. In contrast, the germinated conidia successfully penetrated the cuticle of T. urticae, and within 60 h the fungus colonized the mite's body. Our study demonstrated that although several B. bassiana strains displayed a high virulence in T. urticae there was no evident pathogenicity to phytoseiid mites. These findings support the potential use of entomopathogenic fungus in combination with predatory mites in T. urticae control programs.

  19. Molecular characterization of SAT-2 foot-and-mouth disease virus isolates obtained from cattle during a four-month period in 2001 in Limpopo Province, South Africa.

    Science.gov (United States)

    Phologane, B S; Dwarka, R M; Haydon, D T; Gerber, L J; Vosloo, W

    2008-12-01

    Foot-and-mouth disease (FMD) is an acute, highly contagious viral infection of domestic and wild cloven-hoofed animals. The virus is a single-stranded RNA virus that has a high rate of nucleotide mutation and amino acid substitution. In southern Africa the South African Territories (SAT) 1-3 serotypes of FMD virus are maintained by large numbers of African buffaloes (Syncerus caffer), which provide a potential source of infection for domestic livestock and wild animals. During February 2001, an outbreak of SAT-2 was recorded in cattle in the FMD control zone of South Africa, adjacent to the Kruger National Park (KNP). They had not been vaccinated against the disease since they form the buffer between the vaccination and free zones but in the face of the outbreak, they were vaccinated as part of the control measures to contain the disease. The virus was, however, isolated from some of them on several occasions up to May 2001. These isolates were characterized to determine the rate of genetic change in the main antigenic determinant, the 1 D/2A gene. Nucleotide substitutions at 12 different sites were identified of which five led to amino acid changes. Three of these occurred in known antigenic sites, viz. the GH-loop and C-terminal part of the protein, and two of these have previously been shown to be subject to positive selection. Likelihood models indicated that the ratio of non-synonymous to synonymous changes among the outbreak sequences recovered from cattle was four times higher than among comparable sequences isolated from wildlife, suggesting that the virus may be under greater selective pressure during rapid transmission events.

  20. Offspring sex is not related to maternal allocation of yolk steroids in the lizard Bassiana duperreyi (Scincidae).

    Science.gov (United States)

    Radder, Rajkumar; Ali, Sinan; Shine, Richard

    2007-01-01

    The eggs of birds and reptiles contain detectable levels of several steroid hormones, and experimental application of such steroids can reverse genetically determined sex of the offspring. However, any causal influence of maternally derived yolk steroids on sex determination in birds and reptiles remains controversial. We measured yolk hormones (dihydrotestosterone, testosterone, and 17 beta-estradiol) in newly laid eggs of the montane scincid lizard Bassiana duperreyi. This species is well suited to such an analysis because (1) offspring sex is influenced by incubation temperatures and egg size as well as by sex chromosomes, suggesting that yolk hormones might somehow be involved in the complex pathways of sex determination, and (2) experimental application of either estradiol or fadrozole to such eggs strongly influences offspring sex. We obtained yolk by biopsy, before incubating the eggs at a temperature that produces a 50:50 sex ratio. Yolk steroid levels varied over a threefold range between eggs from different clutches, but there were no significant differences in yolk steroids, or in relative composition of steroids, between eggs destined to become male versus female. Further, yolk steroid concentrations were not significantly related to egg size. Thus, yolk steroid hormones do not appear to play a critical role in sex determination for B. duperreyi.

  1. Interaction between the entomopathogenic fungus, Beauveria bassiana (Ascomycota: Hypocreales and the parasitoid wasp, Aphidius colemani Viereck (Hymenoptera: Braconidae

    Directory of Open Access Journals (Sweden)

    F. Emami

    2013-04-01

    Full Text Available The effect of the entomopathogenic fungus, Beauveria bassiana, on the biological characteristics of Aphidius colemani, a parasitoid of the green peach aphid, Myzus persicae, was studied under laboratory conditions. Third-instar nymphs of green peach aphid were infected with 5/3×105 conidia/mL of B. bassiana, which was determined to be the lethal concentration 50 dose. They were then offered to mated female parasitoids for 24 h at different intervals. Results showed that by prolonging the release intervals of parasitoids, the number of mummies and percent emergence of parasitoids were reduced. Moreover, production of male offspring increased in the F1 generation of parasitoids. The interference of B. bassiana with parasitoid development was also studied by first exposing the aphid hosts to the parasitoids for 24 h and subsequently spraying them with B. bassiana 24, 48, 72, and 96 h after exposure. Results showed that by prolonging fungal spraying intervals, the number of mummies and percent emergence of parasitoids were increased. It appeared that the best time for applying B. bassiana would be three to four days after parasitisation.

  2. The effect of Beauveria bassiana infection on cell mediated and humoral immune response in house fly, Musca domestica L.

    Science.gov (United States)

    Mishra, Sapna; Kumar, Peeyush; Malik, Anushree

    2015-10-01

    Entomopathogenic fungi that manifest infections by overcoming insect's immune response could be a successful control agent for the house fly, Musca domestica L. which is a major domestic, medical, and veterinary pest. In this study, the immune response of house fly to Beauveria bassiana infection was investigated to reveal fundamental aspects of house fly hemocyte biology, such as hemocyte numbers and size, which is poorly understood. The total hemocyte counts (THCs) in B. bassiana-infected house fly showed an initial increase (from 6 to 9 h), followed by subsequent decrease (9 to 12 h) with increase in time of infection. The THCs was slightly greater in infected flies than the non-infected ones. Insight into relative hemocyte counts depicted a significant increase in prohemocyte (PR) and decrease in granulocyte (GR) in infected house flies compared to non-infected ones. The relative cell area of hemocyte cells showed a noticeable increase in PR and intermediate cells (ICs), while a considerable reduction was observed for plasmatocyte (PL) and GR. The considerable variation in relative cell number and cell area in the B. bassiana-infected house flies indicated stress development during infection. The present study highlights changes occurring during B. bassiana invasion to house fly leading to establishment of infection along with facilitation in understanding of basic hemocyte biology. The results of the study is expected to help in better understanding of house fly immune response during fungal infection, so as to assist production of more efficient mycoinsecticides for house fly control using B. bassiana.

  3. Effects of Bacillus thuringiensis toxin Cry1Ac and Beauveria bassiana on Asiatic corn borer (Lepidoptera: Crambidae).

    Science.gov (United States)

    Ma, Xiao-Mu; Liu, Xiao-Xia; Ning, Xia; Zhang, Bo; Han, Fei; Guan, Xiu-Min; Tan, Yun-Feng; Zhang, Qing-Wen

    2008-10-01

    In this study, interactions between Cry1Ac, a toxic crystal protein produced by Bacillus thuringiensis (Berliner), and Beauveria bassiana on the mortality and survival of Ostrinia furnacalis was evaluated in the laboratory. The results showed that Cry1Ac is toxic to O. furnacalis. Not only were larval growth and development delayed, but pupation, pupal weight and adult emergency also decreased when larvae were fed on artificial diet containing purified Cry1Ac toxin. When third instars O. furnacalis were exposed to combination of B. bassiana (1.8x10(5), 1.8x10(6) or 1.8x10(7) conidia ml(-1)) and Cry1Ac, (0.2 or 0.8 microg g(-1)), the effect on mortality was additive, however, the combinations of sublethal concentrations showed antagonism between Cry1Ac (3.2 or 13 microg g(-1)) and B. bassiana (1.8x10(5) or 1.8x10(6) conidia ml(-1)). When neonates were reared on sublethal concentrations of Cry1AC until the third instar, and survivors exposed B. bassiana conidial suspension, such treatments showed additive effect on mortality of O. furnacalis except for the combination of Cry1Ac (0.2 microg g(-1)) and B. bassiana (1.8x10(6) conidia ml(-1)) that showed antagonism.

  4. Suscetibilidade de Oligonychus yothersi (Acari: Tetranychidae ao fungo Beauveria bassiana Susceptibility of Oligonychus yothersi (Acari: Tetranychidae to the fungus Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    Renato Cassol de Oliveira

    2002-03-01

    Full Text Available A cultura da erva-mate (Ilex paraguariensis é uma importante atividade econômica no Sul do Brasil. Com o aumento de consumo nos anos oitenta, cresceram também as áreas de monocultura, gerando condições favoráveis para o aumento populacional de insetos e ácaros fitófagos, entre eles o ácaro vermelho Oligonychus yothersi. Este estudo avaliou a suscetibilidade do ácaro vermelho O. yothersi a vários isolados do fungo Beauveria bassiana. O experimento foi conduzido em Cascavel, PR. Discos foliares de erva-mate (2,2 cm² foram previamente infestados com 20 fêmeas adultas. Alíquotas de 1 mL de cada suspensão de conídios dos diferentes isolados de B. bassiana, ajustadas na concentração de 1,0x10(8 conídios mL-1 foram pulverizadas, separadamente, sobre um conjunto de 5 discos. Os discos foliares foram mantidos flutuando em água destilada, em caixas plásticas (3 cm de diâmetro, sob condições controladas. Diariamente, os ácaros mortos foram transferidos para câmara úmida, para confirmação de mortalidade causada pelo fungo. A mortalidade total variou entre 77 e 98% (6 dias após a aplicação, não permitindo diferenciar os isolados quanto à virulência (P>0,05. Já a mortalidade confirmada variou entre 19 e 75%, permitindo diferenciá-los (PThe Paraguay tea (Ilex paraguariensis crop is an important economical activity in Southern Brazil. With the increase of tea consumption in the 80s, the number of monoculture crops also increased, creating favorable conditions for the development of phytophagous insect and mite populations, such as the red mite Oligonychus yothersi. The objective of this study was to evaluate the susceptibility of the red mite O. yothersi to the several strains of the Beauveria bassiana fungus. The experiments were carried out in Cascavel, PR, Brazil. Leaf disks of Paraguay tea (2.2 cm² were previously infested with 20 adult females. Five disks were inoculated with 1 mL of each conidial suspension of B

  5. [In vitro susceptibilities to levofloxacin and various antibacterial agents of 12,919 clinical isolates obtained from 72 centers in 2007].

    Science.gov (United States)

    Yamaguchi, Keizo; Ohno, Akira; Ishii, Yoshikazu; Tateda, Kazuhiro; Iwata, Morihiro; Kanda, Makoto; Akizawa, Kouji; Shimizu, Chikara; Kon, Shinichirou; Nakamura, Kastushi; Matsuda, Keiko; Tominaga, Makoto; Nakagawa, Takuo; Sugita, Akihiro; Ito, Tatsumi; Kato, Jun; Suwabe, Akira; Yamahata, Kumiko; Kawamura, Chizuko; Tashiro, Hiromi; Horiuchi, Hiroko; Katayama, Yosei; Kondou, Shigemi; Misawa, Shigeki; Murata, Misturu; Kobayashi, Yoshio; Okamoto, Hideyuki; Yamazaki, Kenichiro; Okada, Motoi; Haruki, Kosuke; Kanno, Harushige; Aihara, Masanori; Maesaki, Shigefumi; Hashikita, Giichi; Miyajima, Eiji; Sumitomo, Midori; Saito, Takefumi; Yamane, Nobuo; Kawashima, Chieko; Akiyama, Takahisa; Ieiri, Tamio; Yamamoto, Yoshitaka; Okamoto, Yuki; Okabe, Hidetoshi; Moro, Kunihiko; Shigeta, Masayo; Yoshida, Haruyoshi; Yamashita, Masanobu; Hida, Yukio; Takubo, Takayuki; Kusakabe, Tadashi; Masaki, Hiroya; Heijyou, Hitoshi; Nakaya, Hideo; Kawahara, Kunimitsu; Sano, Reiko; Matsuo, Syuji; Kono, Hisashi; Yuzuki, Yosuke; Ikeda, Norio; Idomuki, Masayo; Soma, Masayuki; Yamamoto, Go; Kinoshita, Syohiro; Kawano, Seiji; Oka, Mikio; Kusano, Nobuchika; Kang, Dongchon; Ono, Junko; Yasujima, Minoru; Miki, Makoto; Hayashi, Masato; Okubo, Syunji; Toyoshima, Syunkou; Kaku, Mitsuo; Sekine, Imao; Shiotani, Joji; Horiuchi, Hajime; Tazawa, Yoko; Yoneyama, Akiko; Kumasaka, Kazunari; Koike, Kazuhiko; Taniguchi, Nobuyuki; Ozaki, Yukio; Uchida, Takashi; Murakami, Masami; Inuzuka, Kazuhisa; Gonda, Hideo; Yamaguchi, Ikuo; fujimoto, Yoshinori; Iriyama, Junji; Asano, Yuko; Genma, Hitoshi; Maekawa, Masato; Yoshimura, Hitoshi; Nakatani, Kaname; Baba, Hisashi; Ichiyama, Satoshi; Fujita, Shinichi; Kuwabara, Masao; Okazaki, Toshiro; Fujiwara, Hiromitsu; Ota, Hiromi; Nagai, Astushi; Fujita, Jun; Negayama, Kiyoshi; Sugiura, Tetsuro; Kamioka, Mikio; Murase, Mitsuharu; Yamane, Nobuhisa; Nakasone, Isamu; Okayama, Akihiko; Aoki, Yosuke; Kusaba, Koji; Nakashima, Yukari; Miyanohara, Hiroaki; Hiramatsu, Kazufumi; Saikawa, Tetsunori; Yanagihara, Katsunori; Matsuda, Junichi; Kohno, Shigeru; Mashiba, Koichi

    2009-08-01

    We have reported in this journal in vitro susceptibilities of clinical isolates to antibiotics every year since 1992. In this paper, we report the results of an analysis of in vitro susceptibilities of 12,919 clinical isolates from 72 centers in Japan to selected antibiotics in 2007 compared with the results from previous years. The common respiratory pathogens, Streptococcus pyogenes, Streptococcus pneumoniae, Moraxella catarrhalis and Haemophilus influenzae maintained a high susceptibility to fluoroquinolones (FQs). The resistance of S. pyogenes to macrolides has been increasing every year and this was especially clear this year. Most strains of Enterobacteriaceae except for Escherichia coli showed a high susceptibility to FQs. Almost 30% of E. coli strains were resistant to FQs and the resistance increased further this year. FQs resistance of methicillin-resistant Staphylococcus aureus (MRSA) was approximately 95% with the exception of 45% for sitafloxacin (STFX). FQs resistance of methicillin-susceptible S. aureus (MSSA) was low at about 10%. FQs resistance of methicillin-resistant coagulase negative Staphylococci (MRCNS) was higher than that of methicillin-susceptible coagulase negative Staphylococci (MSCNS), but it was lower than that of MRSA. However, FQs resistance of MSCNS was higher than that of MSSA. FQs resistance of Enterococcus faecalis was 22.5% to 29.6%, while that of Enterococcusfaecium was more than 85% except for STFX (58.3%). In clinical isolates of Pseudomonas aeruginosa derived from urinary tract infections, FQs resistance was 21-27%, which was higher than that of P. aeruginosa from respiratory tract infections at 13-21%, which was the same trend as in past years. Multidrug resistant strains accounted for 5.6% in the urinary tract and 1.8% in the respiratory tract. Acinetobacter spp. showed high susceptibility to FQs. The carbapenem resistant strains, which present a problem at present, accounted for 2.7%. Neisseria gonorrhoeae showed high

  6. Study on Application of Beauveria bassiana in Controlling Overwintering Dendrolimus tabulaeformis in North Forest%应用球孢白僵菌防治北方越冬油松毛虫的研究

    Institute of Scientific and Technical Information of China (English)

    李永福

    2011-01-01

    为在北方应用白僵菌防治油松毛虫,笔者采用从灵丘县干旱林区分离得到的油松毛虫病原球孢白僵菌菌株Beauveria bassiana CGMCC NO.1573制备的孢子悬浮液和营养型湿菌粉剂,分别在冬季和春季对越冬的油松毛虫幼虫进行防治。结果显示:用2×106个/mL孢子悬浮液的防治效果不好,校正死亡率只有21%~29%;冬季用菌粉防治效果较好,感染率达到70.4%~85.4%;春季施用菌粉15 d~20 d,感染率达47.1%~54.5%.结论认为,在树干基部和枯落物下撒施"白僵菌营养型湿菌粉剂"集中防治越冬幼虫是北方防治油松毛虫的有效方法。%In order to control pine caterpillar effectively in north forest by using Beauveria bassiana,a strain of Beauveria bassiana CGMCC NO.1573,isolated from Dendrolimus tabulaeformis larva from droughty pine forest of Lingqiu County in Shanxi Province,used as spore suspension preparation and nutrition wet bacteria powder,was employed to the control tests in winter and springtime,respectively.The results showed that the control effect was poor by spraying suspension liquid containing spores of 2×106 numbers per mililiter,and the mortality of the Dendrolimus tabulaeformis larvae was only 21%-29%.While the control effect was very good by using the nutrition wet bacteria powder,the infection rate of overwintering larvae reached 70.4%-85.4%.By comparison,the infection rate reached 47.1%-54.5%,within 15-20 days after using the power preparation in spring.So the conclusion is that it is an effective method by broadcasting the nutrition wet bacteria powder preparation of Beauveria bassiana under the litter layer around the trunk base in the pine forest to control gregarious Dendrolimus tabulaeformis larvae in north China.

  7. Selection of optimum conditions of medium acidity and aeration for submerget cultivation of Bacillus thuringiensis and Beauveria bassiana

    Directory of Open Access Journals (Sweden)

    O. A. Dregval

    2010-06-01

    Full Text Available The paper deals with the influence of medium pH and aeration rate on growth and sporulation of Bacillus thuringiensis and Вeauveria bassiana, which are main constituents of the complex microbial insecticide. It was established optimal medium pH for B. thuringiensis – 6.0 and for В. bassiana – 6.0–7.0. The maximum productivity of the studied microorganisms was observed in the same range of aeration – 7– 14 mmol O2/l/h. The selected conditions of cultivation are necessary for the production of complex biological insecticide based on the association of B. thuringiensis and B. bassiana.

  8. The Stress-responsive and Host-oriented Role of Nonribosomal Peptide Synthetases in an Entomopathogenic Fungus, Beauveria bassiana.

    Science.gov (United States)

    Liu, Hang; Xie, Linan; Wang, Jing; Guo, Qiannan; Yang, Shengnan; Liang, Pei; Wang, Chengshu; Lin, Min; Xu, Yuquan; Zhang, Liwen

    2016-11-14

    Beauveria bassiana infects numbers of pest species and is known to produce insecticidal substances, e.g., the nonribosomal peptides (NRPs) beauvericin and bassianolide. However, most NRPs and their biological roles in B. bassiana remain undiscovered. To identify NRPs that potentially contribute to pathogenesis, the 21 predicted NRP synthetases (NRPSs) or NRPS-like proteins of B. bassiana ARSEF2860 were primarily ranked into three functional groups: basic metabolism (7 NRPSs), pathogenicity (12 NRPSs) and unknown function (2 NRPSs). Based on the transcript levels during in vivo growth on diamondback moth (Plutella xylostella, Linnaeus), half of the Group II NRPSs were likely to be involved in infection. Given that the metabolites biosynthesized by these NRPSs remaining to determined, our result underlines the importance of NRPSome in fungal pathogenesis, and will serve as a guide for future genomic mining projects to discover functionally essential and structurally diverse NRPs in fungal genomes.

  9. Quantitative Analysis of Single-Nucleotide Polymorphism for Rapid Detection of TR34/L98H- and TR46/Y121F/T289A-Positive Aspergillus fumigatus Isolates Obtained from Patients in Iran from 2010 to 2014.

    Science.gov (United States)

    Mohammadi, Faezeh; Hashemi, Seyed Jamal; Zoll, Jan; Melchers, Willem J G; Rafati, Haleh; Dehghan, Parvin; Rezaie, Sasan; Tolooe, Ali; Tamadon, Yalda; van der Lee, Henrich A; Verweij, Paul E; Seyedmousavi, Seyedmojtaba

    2015-11-02

    We employed an endpoint genotyping method to update the prevalence rate of positivity for the TR34/L98H mutation (a 34-bp tandem repeat mutation in the promoter region of the cyp51A gene in combination with a substitution at codon L98) and the TR46/Y121F/T289A mutation (a 46-bp tandem repeat mutation in the promoter region of the cyp51A gene in combination with substitutions at codons Y121 and T289) among clinical Aspergillus fumigatus isolates obtained from different regions of Iran over a recent 5-year period (2010 to 2014). The antifungal activities of itraconazole, voriconazole, and posaconazole against 172 clinical A. fumigatus isolates were investigated using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) broth microdilution method. For the isolates with an azole resistance phenotype, the cyp51A gene and its promoter were amplified and sequenced. In addition, using a LightCycler 480 real-time PCR system, a novel endpoint genotyping analysis method targeting single-nucleotide polymorphisms was evaluated to detect the L98H and Y121F mutations in the cyp51A gene of all isolates. Of the 172 A. fumigatus isolates tested, the MIC values of itraconazole (≥16 mg/liter) and voriconazole (>4 mg/liter) were high for 6 (3.5%). Quantitative analysis of single-nucleotide polymorphisms showed the TR34/L98H mutation in the cyp51A genes of six isolates. No isolates harboring the TR46/Y121F/T289A mutation were detected. DNA sequencing of the cyp51A gene confirmed the results of the novel endpoint genotyping method. By microsatellite typing, all of the azole-resistant isolates had genotypes different from those previously recovered from Iran and from the Dutch TR34/L98H controls. In conclusion, there was not a significant increase in the prevalence of azole-resistant A. fumigatus isolates harboring the TR34/L98H resistance mechanism among isolates recovered over a recent 5-year period (2010 to 2014) in Iran. A quantitative assay detecting a single

  10. Expression profiling of Bombyx mori gloverin2 gene and its synergistic antifungal effect with cecropin A against Beauveria bassiana.

    Science.gov (United States)

    Lü, Dingding; Geng, Tao; Hou, Chengxiang; Qin, Guangxing; Gao, Kun; Guo, Xijie

    2017-02-05

    Gloverin2 is a cationic and glycine-rich antimicrobial peptide whose expression can be induced in fat body of silkworm (Bombyx mori) larvae exposed to bacteria. The purpose of this study is to identify the roles of Bombyx mori gloverin2 (Bmgloverin2) during entomopathogenic fungus Beauveria bassiana infection. Fluorescent quantitative real-time PCR analysis indicated that the relative expression level of Bmgloverin2 gene was up-regulated in the silkworm larvae infected by B. bassiana. The cDNA of Bmgloverin2 was cloned from the silkworm by RT-PCR and the DNA segment of the Bmgloverin2 peptide (without signal peptide sequence) was inserted into pCzn1 expression plasmid and expressed in E. coli ArcticExpress (DE3). SDS-PAGE results revealed that soluble recombinant Bmgloverin2 was successfully expressed and purified. Polyclonal antibody against the Bmgloverin2 was successfully produced with the expressed recombinant protein. Western blot analysis indicated that Bmgloverin2 could be detected in the fat body of silkworm larvae infected with B. bassiana, suggesting that the expression of Bmgloverin2 could be induced by B. bassiana infection in silkworm. Antifungal assays indicated that the Bmgloverin2 had a synergistic antifungal activity with B. mori cecropin A (BmCecA) to entomopathogenic fungus B. bassiana both in vitro and in vivo in the silkworm larvae. This is the first report that Bmgloverin2 exhibits synergistic effect with BmCecA in antifungal activity against B. bassiana. The study demonstrates that Bmgloverin2 is an antifungal protein which plays an important role in synergistic antifungal activity with other antimicrobial peptide in silkworm.

  11. Isolation of bacteriophages to multi-drug resistant Enterococci obtained from diabetic foot: A novel antimicrobial agent waiting in the shelf?

    Directory of Open Access Journals (Sweden)

    C S Vinodkumar

    2011-01-01

    Full Text Available Introduction: While foot infections in persons with diabetes are initially treated empirically, therapy directed at known causative organisms may improve the outcome. Many studies have reported on the bacteriology of diabetic foot infections (DFIs, but the results have varied and have often been contradictory. The purpose of the research work is to call attention to a frightening twist in the antibiotic-resistant Enterococci problem in diabetic foot that has not received adequate attention from the medical fraternity and also the pharmaceutical pipeline for new antibiotics is drying up. Materials and Methods: Adult diabetic patients admitted for lower extremity infections from July 2008 to December 2009 in the medical wards and intensive care unit of medical teaching hospitals were included in the study. The extent of the lower extremity infection on admission was assessed based on Wagner′s classification from grades I to V. Specimens were collected from the lesions upon admission prior to the initiation of antibiotic therapy or within the first 48 h of admission. Results: During the 18-month prospective study, 32 strains of Enterococcus spp. (26 Enterococcus faecalis and 06 E. faecium were recovered. Antibiotic sensitivity testing was done by Kirby-Bauer′s disk diffusion method. Isolates were screened for high-level aminoglycoside resistance (HLAR. A total of 65.6% of Enterococcus species showed HLAR. Multidrug resistance and concomitant resistance of HLAR strains to other antibiotics were quite high. None of the Enterococcus species was resistant to vancomycin. Conclusion: Multidrug-resistant Enterococci are a real problem and continuous surveillance is necessary. Today, resistance has rendered most of the original antibiotics obsolete for many infections, mandating the development of alternative anti-infection modalities. One of such alternatives stemming up from an old idea is the bacteriophage therapy. In the present study, we could

  12. Toxicity of fifteen strains of Beauveria bassiana against Aphis glycines%球孢白僵菌对大豆蚜毒力的室内生物学测定

    Institute of Scientific and Technical Information of China (English)

    刘兴磊; 张正坤; 徐文静; 刘影; 王金刚; 李启云

    2011-01-01

    The toxicity of fifteen strains of the entomopathogenic fungus Beauveria bassiana ( Bals. ) Vuill. To Aphis glycines Matsumura was determined. All tested strains were toxic to A. Glycines and obvious differences in the mortality rates achieved by the different strains were positively correlated with spore concentration and treatment time. Strain S14 - X - 1 had the highest amount of toxic isolates.%对来源于吉林省不同地区的15株球孢白僵菌Beauveria bassiana( Bals.) Vuill.菌种对大豆蚜Aphisglycines Matsumura毒力进行了室内生物学测定.所有供试菌株对大豆蚜虫均有一定的致病性,杀虫效率差异明显,杀虫效率与孢子浓度和作用时间均呈正相关.通过对大豆蚜毒力室内生物学测定结果的综合分析,菌株S14-X-1为毒力较高菌株,具有进一步研究应用价值.

  13. Molecular characterization and pathogenicity of fungal isolates for use against the small hive beetle (Aethina tumida)

    Science.gov (United States)

    The analysis of DNA sequences from fungal pathogens obtained from cadavers of the small hive beetle (SHB) collected from several apiaries in Florida revealed a mixture of saprobes and two potential primary entomopathogens, Metarhizium anisopliae and Beauveria bassiana. Spray tower bioassays indicate...

  14. The identification of a bacterial strain BGI-1 isolated from the intestinal flora of Blattella germanica, and its anti-entomopathogenic fungi activity.

    Science.gov (United States)

    Huang, Y H; Wang, X J; Zhang, F; Huo, X B; Fu, R S; Liu, J J; Sun, W B; Kang, D M; Jing, X

    2013-02-01

    A bacterial strain BGI-1 was isolated from the gut of German cockroaches (Blattella germanica L.) and was identified as Bacillus subtilis based on 16S rDNA sequence and morphological, physiological, and biochemical characters. The strain BGI-1 inhibited the growth of Beauveria bassiana; the diameter of the inhibition zone exceeded 30 mm. Vesicles were observed in B. bassiana hyphae on the edge of the inhibition zone. Fermentation of BGI-1 reduced the conidial germination rate by 12%. Further studies demonstrated that B. bassiana infections in German cockroaches orally treated with the extracts of BGI-1 fermentation were significantly weakened. Cumulative mortality rate was 49.5% in the treatment group at the 20 d, while that of the control group was 62.3%. The study intends to understand the relationship between the intestinal flora and the cockroach. Those microbes with anti-entomopathogenic fungi activity might contribute to resisting the infection of pathogenic fungi.

  15. 紫外线诱变选育苜蓿叶象甲高毒力白僵菌%Screening Highly Virulent Strains of Beauveria bassiana to Hypera postica Gyllenhal by UV Mutagenesis

    Institute of Scientific and Technical Information of China (English)

    韩慧; 吴小虎; 李益; 李克梅; 赵莉

    2011-01-01

    [Objective] The purpose of this program was to screen out highly virulent strains of Beauveria bassiana to Hypera postica Cyllenhal. [Method] Ultraviolet irradiating was used to irradiate Beauveria bassiana strain for 10 min and 30 min in order to select high chitinase mutant, and then 3 stage larvae of Hypera postica underwent the virulence test with the Beauveria bassiana before and after UV mutagenesis. [Result]The results of the test indicated that corrected mortality of the stain Bb - 10 - 12 was 100% , corrected mortality of the stain Bb -30- 14 was 77.78%, and corrected mortality of wild type strain was 62.97% in 9 days later. [Conclusion] A more virulent strain Bb - 10 - 12 of Beauveria bassiana to Hypera postica was obtained by UV mutagenesis.%[目的]选育出对苜蓿叶象甲高毒力的白僵菌.[方法]利用紫外线对前期筛选出的白僵菌菌株照射10和30min进行诱变,选出高几丁质酶突变株,再用诱变前后的白僵菌对苜蓿叶象甲3龄幼虫进行毒力测定.[结果]用诱变前后的白僵菌对苜蓿叶象甲3龄幼虫室内毒力测定:处理9d后,诱导菌株Bb- 10- 12校正死亡率为100%,Bb - 30 - 14校正死亡率为77.78%,而朱经紫外线照射的野生型菌株校正死亡率为62.97%.[结论]通过室内毒力测定法,获得了经10 min紫外线照射诱变对苜蓿叶象甲更高毒力的1株白僵菌菌株.

  16. Preparation, characterization, and insecticidal activity evaluation of three different formulations of Beauveria bassiana against Musca domestica.

    Science.gov (United States)

    Mishra, Sapna; Kumar, Peeyush; Malik, Anushree

    2013-10-01

    Three formulations; bait, encapsulation, and emulsion of Beauveria bassiana were prepared and evaluated for their insecticidal activity in simulated field settings. Tea waste-based bait formulation of B. bassiana showed 100% mortality (within 72 h) in lab assay against adult houseflies. In field assay using traps, 65% relative entrapment and 100 % mortality (within 60 h) of entrapped flies was observed. Although the bait formulation was low cost and easy to prepare and transport, its storage ability was limited. Hence, more advanced formulations in form of encapsulation and emulsion was attempted. Encapsulated B. bassiana conidia (using skimmed milk powder, polyvinyl pyrrolidone K-90 and glucose as additives) showed 100% conidial germination and retained 78% conidial viability, even after storage for 12 months at 30 °C. Encapsulated product showed 54.8% (freshly prepared) and 30.6 % (after 12-months storage) mortality of housefly larvae in a simulated field condition. Emulsion formulation was prepared by using Tween 20 as surfactant with seven vegetable oils: soybean, rapeseed, sunflower, olive, castor, til, and linseed. Emulsion with linseed oil showing maximum conidial germination (94%) was evaluated for shelf life and pathogenecity against housefly larvae. Shelf life analysis of emulsion revealed 28% conidial germination and 19.9% housefly larval mortality after 12 months of storage as opposed to 94% conidial germination and 51.7% of larval mortality with fresh product. Significant increase in shelf × targeted application of formulation is expected to increase its mass applicability for housefly control. Also, the variability among products presents diverse opportunities for commercialization.

  17. Improved shelf life of dried Beauveria bassiana blastospores using convective drying and active packaging processes.

    Science.gov (United States)

    Mascarin, Gabriel Moura; Jackson, Mark A; Behle, Robert W; Kobori, Nilce N; Júnior, Ítalo Delalibera

    2016-10-01

    The yeast form (blastospore) of the dimorphic insect-pathogenic fungus Beauveria bassiana can be rapidly produced using liquid fermentation methods but is generally unable to survive rapid dehydration processes or storage under non-refrigerated conditions. In this study, we evaluated the influence of two convective drying methods, various modified atmosphere packaging systems, and storage temperatures on the desiccation tolerance, storage stability, and virulence of blastospores of B. bassiana ESALQ 1432. All blastospore formulations were dried to <5 % water content equivalent to aw < 0.3. The viability of B. bassiana blastospores after air drying and spray drying was greater than 80 %. Vacuum-packaged blastospores remained viable longer when stored at 4 °C compared with 28 °C with virtually no loss in viability over 9 months regardless the drying method. When both oxygen and moisture scavengers were added to sealed packages of dried blastospore formulations stored at 28 °C, viability was significantly prolonged for both air- and spray-dried blastospores. The addition of ascorbic acid during spray drying did not improve desiccation tolerance but enhanced cell stability (∼twofold higher half-life) when stored at 28 °C. After storage for 4 months at 28 °C, air-dried blastospores produced a lower LC80 and resulted in higher mortality to whitefly nymphs (Bemisia tabaci) when compared with spray-dried blastospores. These studies identified key storage conditions (low aw and oxygen availability) that improved blastospore storage stability at 28 °C and will facilitate the commercial development of blastospores-based bioinsecticides.

  18. Sublethal Effects of Beauveria bassiana (Ascomycota: Hypocreales) on Life Table Parameters of Frankliniella occidentalis (Thysanoptera: Thripidae).

    Science.gov (United States)

    Zhang, Tao; Reitz, Stuart R; Wang, Haihong; Lei, Zhongren

    2015-06-01

    We assessed effects of parental exposure to Beauveria bassiana on life history traits of subsequent generations of western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). Progeny from individuals that survived fungal exposure as second instars had significantly shorter egg stages, but longer prepupal development times than corresponding untreated controls. However, survivorship to adulthood of these progeny groups did not differ. Although fecundities of the parental types did not differ, the sex ratio of progeny from fungal-treated parents was male-biased, whereas sex ratio of progeny from untreated control parents was even. We calculated life table parameters for the progeny and found that all parameters, except for generation time, were significantly less for the progeny of fungal-treated parents than for progeny of untreated parents. The intrinsic rate of increase, finite rate of increase, net reproductive rate, mean generation time, and gross reproductive rate were 0.199 d(-1), 1.229 d(-1), 21.84, 15.48 d, and 27.273, respectively, for progeny of treated thrips, and 0.266 d(-1), 1.316 d(-1), 52.540, 14.92 d, and 70.64, respectively, for progeny of control thrips. Consequently, population projections demonstrated that offspring of parents exposed to B. bassiana would increase their population more slowly than those from untreated parents. These results demonstrate that B. bassiana has sublethal effects that reduce the reproductive success of F. occidentalis and these effects should be taken into account when evaluating its use in management programs for F. occidentalis.

  19. Antimicrobial resistance profile of Staphylococcus aureus isolates obtained from skin and soft tissue infections of outpatients from a university hospital in Recife - PE, Brazil*

    Science.gov (United States)

    Caraciolo, Fabiana Beserra; Maciel, Maria Amélia Vieira; dos Santos, Josemir Belo; Rabelo, Marcelle Aquino; Magalhães, Vera

    2012-01-01

    BACKGROUND Staphylococcus aureus has a notable ability to acquire resistance to antibiotics, and methicillin resistance represents a growing public health problem. Methicillin-resistant S. aureus (MRSA) has also become important outside the hospital environment, particularly in the United States. In Brazil, since 2005, cases of community skin infections caused by MRSA have been reported, but resistance studies involving outpatients are scarce. OBJECTIVE To know the resistance profile of S. aureus involved in skin and soft tissue infections of patients seen at the Dermatology outpatient clinic of a university hospital in Recife, Pernambuco State, northeastern Brazil. METHODS Prospective study involving 30 patients with skin and soft tissue infections, seen at the Dermatology outpatient clinic from May until November 2011. To evaluate the susceptibility of S. aureus to antibiotics, the disk diffusion method and oxacillin screening agar were used. RESULTS From a total of 30 samples of skin lesions, 19 (63%) had positive culture for S. aureus. The following resistance patterns of S. aureus were observed: penicillin, 95%; tetracycline, 32%; erythromycin, 21%; gentamicin, 16%; cefoxitin, 11%; oxacillin, 11%; trimethoprim-sulfamethoxazole, 11%; chloramphenicol, 11%; clindamycin, 5% ; and ciprofloxacin, 0%. One of the identified MRSA was obtained from a patient without risk factors for its acquisition, and was resistant, beyond to the beta-lactams, only to tetracycline. CONCLUSIONS With regard to the resistance patterns of S. aureus, resistances to tetracycline, erythromycin and gentamicin were the highest. It was documented, for the first time in Pernambuco, a case of skin infection caused by community-associated MRSA. PMID:23197204

  20. rRNA sequence comparison of Beauveria bassiana, Tolypocladium cylindrosporum, and Tolypocladium extinguens.

    Science.gov (United States)

    Rakotonirainy, M S; Dutertre, M; Brygoo, Y; Riba, G

    1991-01-01

    Five strains of Tolypocladium cylindrosporum, one strain of Tolypocladium extinguens, and nine strains of Beauveria bassiana were analyzed using a rapid rRNA sequencing technique. The sequences of two highly variable domains (D1 and D2) located at the 5' end of the 28S-like rRNA molecule were determined. The phylogenetic tree computed from the absolute number of nucleotide differences shows the separation between the genus Beauveria and the genus Tolypocladium and points out that T. cylindrosporum and T. extinguens probably do not belong to the same genus.

  1. 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

  2. Cloning and expression in Saccharomyces cerevisiae of chit2 gene from Beauveria bassiana

    Institute of Scientific and Technical Information of China (English)

    SONG Jin-zhu; YANG Xiao-xue; WANG Yun; YANG Qian

    2009-01-01

    To study recycled trashes from shrimps and crabs in the sea through chitinase secreted by microor-ganisms, the chitinase gene chit2 was cloned and sequenced from Beauveria bassiana by the polymerase chain reaction (PCR), and was ligated into the yeast expression vector pYES2. The expression vector plasmid was transformed into Saccharomyces cerevisiae H158. Gene expression took place upon induction with 2% galac-tose. The measurement of enzyme activity shows that the expression production can be expressed in active forms and secreted to the medium. The enzyme activity approaches the peak of 0. 63 U/mL when the culture time is 36 h.

  3. Insight into the feeding behavior of predatory mites on Beauveria bassiana, an arthropod pathogen.

    Science.gov (United States)

    Wu, Shengyong; Zhang, Ye; Xu, Xuenong; Lei, Zhongren

    2016-04-04

    Interactions between fungal entomopathogens and pest predators are particularly relevant in control of agricultural insect pests. In a laboratory study, we confirmed that the predatory mite, Neoseiulus barkeri, exhibited feeding behavior on the entomopathogenic fungus Beauveria bassiana conidia through DNA extracts. Using transmission electron microscopy, we determined that the majority of conidia found in the mite gut tended to dissolve within 24 h post ingestion, suggesting that the conidia had probably lost their viability. To our knowledge this is the first report of feeding behavior of phytoseiid mites on entomopathogenic fungus. The findings expand our knowledge of fungus-predator interactions.

  4. Susceptibility of the Parasitoid Phymastichus coffea LaSalle (Hymenoptera: Eulophidae) to Beauveria bassiana under laboratory conditions; Susceptibilidad del parasitoide Phymastichus coffea LaSalle (Hymenoptera:Eulophidae) a Beauveria bassiana en condiciones de laboratorio

    Energy Technology Data Exchange (ETDEWEB)

    Castillo, Alfredo; Gomez, Jaime; Infante, Francisco [El Colegio de la Frontera Sur (ECOSUR), Chiapas (Mexico). Dept. de Entomologia Tropical], e-mail: acastill@ecosur.mx, e-mail: jgomez@ecosur.mx, e-mail: finfante@ecosur.mx; Vega, Fernando E. [United States Department of Agriculture (USDA), Beltsville, MD (United States). Agricultural Research Service. Sustainable Perennial Crops Lab.], e-mail: fernando.vega@ars.usda.gov

    2009-09-15

    The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae), is the most important coffee pest worldwide. Beauveria bassiana is a generalist entomopathogenic fungus widely used by coffee farmers to control this pest and Phymastichus coffea LaSalle (Hymenoptera: Eulophidae) is an African endo parasitoid of H. hampei adults, recently imported to several Latin American and Caribbean countries to aid in the coffee berry borer control. The objective of this study was to determine if B. bassiana is detrimental to P. coffea. The susceptibility of the parasitoid was evaluated in terms of adult survivorship, mean lethal concentration (LC{sub 50}), mean lethal time (LT{sub 50}), reproduction and immature mortality. The main effect of the fungus resulted in reduction of adult longevity and mortality of 100% for immature stages of this parasitoid. The LC{sub 50} for adults was 0.11% equivalent to 9.53 x 10{sup 7} conidia/ml of B. bassiana and a LT{sub 50} of 29.4 h, equivalent to reduction of 22% of its normal longevity as an adult. P. coffea was capable of disseminating spores of B. bassiana to non-infected H. hampei adults, which could indirectly cause the death of its own progeny. These results could be valuable when considering the use of both organisms in the field, especially in an integrated pest management program. (author)

  5. Phylogeny of type I polyketide synthases (PKSs) in fungal entomopathogens and expression analysis of PKS genes in Beauveria bassiana BCC 2660.

    Science.gov (United States)

    Punya, Juntira; Swangmaneecharern, Pratchya; Pinsupa, Suparat; Nitistaporn, Pornpen; Phonghanpot, Suranat; Kunathigan, Viyada; Cheevadhanarak, Supapon; Tanticharoen, Morakot; Amnuaykanjanasin, Alongkorn

    2015-06-01

    Entomopathogenic fungi are able to invade and kill insects. Various secondary metabolites can mediate the interaction of a fungal pathogen with an insect host and also help the fungus compete with other microbes. Here we screened 23 isolates of entomopathogenic fungi for polyketide synthase (PKS) genes and amplified 72 PKS gene fragments using degenerate PCR. We performed a phylogenetic analysis of conserved ketosynthase and acyltransferase regions in these 72 sequences and 72 PKSs identified from four insect fungal genome sequences. The resulting genealogy indicated 47 orthologous groups with 99-100 % bootstrap support, suggesting shared biosynthesis of identical or closely related compounds from different fungi. Three insect-specific groups were identified among the PKSs in reducing clades IIa, IIb, and III, which comprised PKSs from 12, 9, and 30 fungal isolates, respectively. A IIa-IIb pair could be found in seven fungi. Expression analyses revealed that eleven out of twelve PKS genes identified in Beauveria bassiana BCC 2660 were expressed in culture. PKS genes from insect-specific clades IIa and IIb were expressed only in insect-containing medium, while others were expressed only in PDB or in CYB, PDB and SDY. The data suggest the potential production of several polyketides in culture.

  6. Ação do fungo Beauveria bassiana, isolado 986, sobre o ciclo biológico do cascudinho Alphitobius diaperinus em laboratório Action of the fungus Beauveria bassiana, strain 986, over the biologic cycle of the Alphitobius diaperinus beetle in laboratory

    Directory of Open Access Journals (Sweden)

    Aleksandro Schafer da Silva

    2006-12-01

    Full Text Available O presente trabalho foi realizado para verificar a eficácia do fungo Beauveria bassiana em todas as etapas de desenvolvimento de Alphitobius diaperinus e assim saber em que fase ele é mais susceptível ao controle biológico. O fungo B. bassiana, isolado 986, foi testado em duas concentrações (3,4 X 10(6 e 3,4 X 10(8 conídios ml-1 mais um grupo controle. Foram realizadas duas leituras, uma sete e outra quatorze dias após a aplicação dos fungos. Na concentração 3,4 X 10(6, 54% dos ovos tratados estavam inférteis, ocorrendo 54% de mortalidade de larvas tratadas nos estádios I, II e III, 22,5% nos estádios IV, V e VI, 9,5% nos estádios VII e VIII e 24,5% de mortalidade das pupas. Na concentração 3,4 X 10(8, 66,8% dos ovos tratados estavam inviáveis, ocorrendo mortalidade de 56% das larvas tratadas nos estádios I, II e III, 34% nos estádios IV, V e VI, 24,5% nos estádios VII e VIII e 49,5% das pupas. Já no grupo controle, 13,3% dos ovos estavam inviáveis, ocorrendo mortalidade de 10% das larvas nos estádios I, II e III, 4% nos estádios IV, V e VI e 0% de mortalidade nos estádios VII, VIII e em pupas. Sobre o cascudinho adulto, o fungo não teve efeito nocivo em nenhum grupo analisado. Com base nesses resultados, observou-se que o fungo nas concentrações testadas apresentou efeito nocivo às fases do ciclo biológico do cascudinho, com exceção do inseto adulto.The present research was carried out to verify the Beauveria bassiana fungus efficacy in all development phases of Alphitobius diaperinus and to know in which phase it is more susceptible for biologic control. The B. bassiana, 986-isolated fungus, was tested in two different concentrations (3.4 x 10(6 and 3.4 x 10(8 conidium ml-1 and a control group. Two readings were realized, one after 7 days of fungus application and the other after 14 days. At concentration of 3.4 x 10(6 conidium ml-1, 54% of treated eggs were infertile, with larva death of 54% at I, II and III

  7. CRISPR/Cas9-mediated efficient genome editing via blastospore-based transformation in entomopathogenic fungus Beauveria bassiana

    Science.gov (United States)

    Chen, Jingjing; Lai, Yiling; Wang, Lili; Zhai, Suzhen; Zou, Gen; Zhou, Zhihua; Cui, Chunlai; Wang, Sibao

    2017-01-01

    Beauveria bassiana is an environmentally friendly alternative to chemical insecticides against various agricultural insect pests and vectors of human diseases. However, its application has been limited due to slow kill and sensitivity to abiotic stresses. Understanding of the molecular pathogenesis and physiological characteristics would facilitate improvement of the fungal performance. Loss-of-function mutagenesis is the most powerful tool to characterize gene functions, but it is hampered by the low rate of homologous recombination and the limited availability of selectable markers. Here, by combining the use of uridine auxotrophy as recipient and donor DNAs harboring auxotrophic complementation gene ura5 as a selectable marker with the blastospore-based transformation system, we established a highly efficient, low false-positive background and cost-effective CRISPR/Cas9-mediated gene editing system in B. bassiana. This system has been demonstrated as a simple and powerful tool for targeted gene knock-out and/or knock-in in B. bassiana in a single gene disruption. We further demonstrated that our system allows simultaneous disruption of multiple genes via homology-directed repair in a single transformation. This technology will allow us to study functionally redundant genes and holds significant potential to greatly accelerate functional genomics studies of B. bassiana. PMID:28368054

  8. Acoustic assessment of Beauveria bassiana (Hypocreales: Clavicipitaceae) effects on Rhynchophorus ferrugineus (Coleoptera: Dryophthoridae) larval activity and mortality

    Science.gov (United States)

    Rhynchophorus ferrugineus (Olivier) (Coleoptera: Dryophthoridae), the red palm weevil, is an economically important palm tree pest in subtropical regions of the world. Previous studies have shown that R. ferrugineus can be infected and killed by the entomopathogenic fungus, Beauveria bassiana. Howev...

  9. Synergistic interaction between the fungus Beauveria bassiana and desiccant dusts applied against poultry red mites (Dermanyssus gallinae)

    DEFF Research Database (Denmark)

    Steenberg, Tove; Kilpinen, Ole Østerlund

    2014-01-01

    , but these fungi often take several days to kill mites. Laboratory experiments were carried out to study the efficacy of 3 types of desiccant dusts, the fungus Beauveria bassiana and combinations of the two control agents against D. gallinae. There was significant synergistic interaction between each...

  10. 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.

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

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    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

  12. Metabolic profiles and free radical scavenging activity of Cordyceps bassiana fruiting bodies according to developmental stage.

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    Sun-Hee Hyun

    Full Text Available The metabolic profiles of Cordyceps bassiana according to fruiting body developmental stage were investigated using gas chromatography-mass spectrometry. We were able to detect 62 metabolites, including 48 metabolites from 70% methanol extracts and 14 metabolites from 100% n-hexane extracts. These metabolites were classified as alcohols, amino acids, organic acids, phosphoric acids, purine nucleosides and bases, sugars, saturated fatty acids, unsaturated fatty acids, or fatty amides. Significant changes in metabolite levels were found according to developmental stage. Relative levels of amino acids, purine nucleosides, and sugars were higher in development stage 3 than in the other stages. Among the amino acids, valine, isoleucine, lysine, histidine, glutamine, and aspartic acid, which are associated with ABC transporters and aminoacyl-tRNA biosynthesis, also showed higher levels in stage 3 samples. The free radical scavenging activities, which were significantly higher in stage 3 than in the other stages, showed a positive correlation with purine nucleoside metabolites such as adenosine, guanosine, and inosine. These results not only show metabolic profiles, but also suggest the metabolic pathways associated with fruiting body development stages in cultivated C. bassiana.

  13. Chemical Constituents Identified from Fruit Body of Cordyceps bassiana and Their Anti-Inflammatory Activity.

    Science.gov (United States)

    Suh, Wonse; Nam, Gyeongsug; Yang, Woo Seok; Sung, Gi-Ho; Shim, Sang Hee; Cho, Jae Youl

    2017-03-01

    Cordyceps bassiana is one of Cordyceps species with anti-oxidative, anti-cancer, anti-inflammatory, anti-diabetic, anti-obesity, anti-angiogenic, and anti-nociceptive activities. This mushroom has recently demonstrated to have an ability to reduce 2,4-dinitrofluorobenzene-induced atopic dermatitis symptoms in NC/Nga mice. In this study, we further examined phytochemical properties of this mushroom by column chromatography and HPLC analysis. By chromatographic separation and spectroscopic analysis, 8 compounds, such as 1,9-dimethylguanine (1), adenosine (2), uridine (3), nicotinamide (4), 3-methyluracil (5), 1,7-dimethylxanthine (6), nudifloric acid (7), and mannitol (8) were identified from 6 different fractions and 4 more subfractions. Through evaluation of their anti-inflammatory activities using reporter gene assay and mRNA analysis, compound 1 was found to block luciferase activity induced by NF-κB and AP-1, suppress the mRNA levels of cyclooxygenase (COX)-2 and tumor necrosis factor (TNF)-α. Therefore, our data strongly suggests that compound 1 acts as one of major principles in Cordyceps bassiana with anti-inflammatory and anti-atopic dermatitis activities.

  14. Myco-synthesis of silver nanoparticles using Beauveria bassiana against dengue vector, Aedes aegypti (Diptera: Culicidae).

    Science.gov (United States)

    Banu, A Najitha; Balasubramanian, C

    2014-08-01

    The efficacy of silver synthesized biolarvicide with the help of entomopathogenic fungus, Beauveria bassiana, was assessed against the different larval instars of dengue vector, Aedes aegypti. The silver nanoparticles were observed and characterized by a scanning electron microscope (SEM) and energy-dispersive X-ray (EDX). A surface plasmon resonance band was observed at 420 nm in UV-vis spectrophotometer. The characterization was confirmed by shape (spherical), size 36.88-60.93 nm, and EDX spectral peak at 3 keV of silver nanoparticles. The synthesized silver nanoparticles have been tested against the different larval instars of Ae. aegypti at different concentrations for a period of 24 h. Ae. aegypti larvae were found more susceptible to the synthesized silver nanoparticles. The LC50 and LC90 values are 0.79 and 1.09 ppm with respect to the Ae. aegypti treated with B. bassiana (Bb) silver nanoparticles (AgNPs). First and second instar larvae of Ae. aegypti have shown cent percent mortality while third and fourth instars found 50.0, 56.6, 70.0, 80.0, and 86.6 and 52.4, 60.0, 68.5, 76.0, and 83.3% mortality at 24 h of exposure in 0.06 and 1.00 ppm, respectively. It is suggested that the entomopathogenic fungus synthesized silver nanoparticles would be appropriate for environmentally safer and greener approach for new leeway in vector control strategy through a biological process.

  15. Metabolic Profiles and Free Radical Scavenging Activity of Cordyceps bassiana Fruiting Bodies According to Developmental Stage

    Science.gov (United States)

    Hyun, Sun-Hee; Lee, Seok-Young; Sung, Gi-Ho; Kim, Seong Hwan; Choi, Hyung-Kyoon

    2013-01-01

    The metabolic profiles of Cordyceps bassiana according to fruiting body developmental stage were investigated using gas chromatography-mass spectrometry. We were able to detect 62 metabolites, including 48 metabolites from 70% methanol extracts and 14 metabolites from 100% n-hexane extracts. These metabolites were classified as alcohols, amino acids, organic acids, phosphoric acids, purine nucleosides and bases, sugars, saturated fatty acids, unsaturated fatty acids, or fatty amides. Significant changes in metabolite levels were found according to developmental stage. Relative levels of amino acids, purine nucleosides, and sugars were higher in development stage 3 than in the other stages. Among the amino acids, valine, isoleucine, lysine, histidine, glutamine, and aspartic acid, which are associated with ABC transporters and aminoacyl-tRNA biosynthesis, also showed higher levels in stage 3 samples. The free radical scavenging activities, which were significantly higher in stage 3 than in the other stages, showed a positive correlation with purine nucleoside metabolites such as adenosine, guanosine, and inosine. These results not only show metabolic profiles, but also suggest the metabolic pathways associated with fruiting body development stages in cultivated C. bassiana. PMID:24058459

  16. Evolutionary diversification of the lizard genus Bassiana (Scincidae across Southern Australia.

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    Sylvain Dubey

    Full Text Available BACKGROUND: Relatively recent (Plio-Pleistocene climatic variations had strong impacts on the fauna and flora of temperate-zone North America and Europe; genetic analyses suggest that many lineages were restricted to unglaciated refuges during this time, and have expanded their ranges since then. Temperate-zone Australia experienced less severe glaciation, suggesting that patterns of genetic structure among species may reflect older (aridity-driven divergence events rather than Plio-Pleistocene (thermally-mediated divergences. The lizard genus Bassiana (Squamata, Scincidae contains three species that occur across a wide area of southern Australia (including Tasmania, rendering them ideally-suited to studies on the impact of past climatic fluctuations. METHODOLOGY/PRINCIPAL FINDINGS: We performed molecular phylogenetic and dating analyses using two partial mitochondrial genes (ND2 and ND4 of 97 samples of Bassiana spp. Our results reveal a pattern of diversification beginning in the Middle Miocene, with intraspecific diversification arising from 5.7 to 1.7 million years ago in the Upper Miocene-Lower Pleistocene. CONCLUSIONS/SIGNIFICANCE: In contrast to the temperate-zone Northern Hemisphere biota, patterns of evolutionary diversification within southern Australian taxa appear to reflect geologically ancient events, mostly relating to east-west discontinuities imposed by aridity rather than (as is the case in Europe and North America relatively recent recolonisation of northern regions from unglaciated refugia to the south.

  17. Beauveria bassiana Lipase A expressed in Komagataella (Pichia) pastoris with potential for biodiesel catalysis

    Science.gov (United States)

    Vici, Ana C.; da Cruz, Andrezza F.; Facchini, Fernanda D. A.; de Carvalho, Caio C.; Pereira, Marita G.; Fonseca-Maldonado, Raquel; Ward, Richard J.; Pessela, Benevides C.; Fernandez-Lorente, Gloria; Torres, Fernando A. G.; Jorge, João A.; Polizeli, Maria L. T. M.

    2015-01-01

    Lipases (EC 3.1.1.3) comprise a biotechnologically important group of enzymes because they are able to catalyze both hydrolysis and synthesis reactions, depending on the amount of water in the system. One of the most interesting applications of lipase is in the biofuel industry for biodiesel production by oil and ethanol (or methanol) transesterification. Entomopathogenic fungi, which are potential source of lipases, are still poorly explored in biotechnological processes. The present work reports the heterologous expression and biochemical characterization of a novel Beauveria bassiana lipase with potential for biodiesel production. The His-tagged B. bassiana lipase A (BbLA) was produced in Komagataella pastoris in buffered methanol medium (BMM) induced with 1% methanol at 30°C. Purified BbLA was activated with 0.05% Triton X-100 and presented optimum activity at pH 6.0 and 50°C. N-glycosylation of the recombinant BbLA accounts for 31.5% of its molecular weight. Circular dichroism and molecular modeling confirmed a structure composed of α-helix and β-sheet, similar to α/β hydrolases. Immobilized BbLA was able to promote transesterification reactions in fish oil, demonstrating potential for biodiesel production. BbLA was successfully produced in K. pastoris and shows potential use for biodiesel production by the ethanolysis reaction. PMID:26500628

  18. Proteins in the Cocoon of Silkworm Inhibit the Growth of Beauveria bassiana

    Science.gov (United States)

    Zhang, Yan; Li, Youshan; Liu, Huawei; Xia, Qingyou; Zhao, Ping

    2016-01-01

    Silk cocoons are composed of fiber proteins (fibroins) and adhesive glue proteins (sericins), which provide a physical barrier to protect the inside pupa. Moreover, other proteins were identified in the cocoon silk, many of which are immune related proteins. In this study, we extracted proteins from the silkworm cocoon by Tris-HCl buffer (pH7.5), and found that they had a strong inhibitory activity against fungal proteases and they had higher abundance in the outer cocoon layers than in the inner cocoon layers. Moreover, we found that extracted cocoon proteins can inhibit the germination of Beauveria bassiana spores. Consistent with the distribution of protease inhibitors, we found that proteins from the outer cocoon layers showed better inhibitory effects against B. bassiana spores than proteins from the inner layers. Liquid chromatography-tandem mass spectrometry was used to reveal the extracted components in the scaffold silk, the outermost cocoon layer. A total of 129 proteins were identified, 30 of which were annotated as protease inhibitors. Protease inhibitors accounted for 89.1% in abundance among extracted proteins. These protease inhibitors have many intramolecular disulfide bonds to maintain their stable structure, and remained active after being boiled. This study added a new understanding to the antimicrobial function of the cocoon. PMID:27032085

  19. PATOGENICIDADE DE BEAUVERIA BASSIANA (BALS. VUILL. AO ÁCARO Tetranychus urticae KOCH

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    Tamai Marco Antonio

    1999-01-01

    Full Text Available O estudo da patogenicidade de Beauveria bassiana (Bals. Vuill. ao ácaro Tetranychus urticae Koch foi desenvolvido a 25 ± 2°C, 70 ± 5% UR e 12 horas de fotofase, utilizando-se fêmeas recém-emergidas. Os ácaros foram mantidos durante seis dias em discos de folha de Canavalia ensiformis (L. DC. inoculadas com B. bassiana, isolado 447, nas concentrações: 5x106, 1x107, 5x107, 1x108, 5x108 e 1x109 conídios/ml. Este isolado mostrou-se patogênico a T. urticae, apresentando aumento nos valores das mortalidades acumuladas (total, corrigida e confirmada à medida que a suspensão de conídios se tornou mais concentrada. Nas seis concentrações testadas, os valores de mortalidade corrigida ao sexto dia, foram inferiores a 50%, sendo observada apenas na concentração de 1x109 conídios/ml, mortalidade total superior a 50%.

  20. Biological Control of Aphid Using Fungal Culture and Culture Filtrates of Beauveria bassiana.

    Science.gov (United States)

    Kim, Jeong Jun; Jeong, Gayoung; Han, Ji Hee; Lee, Sangyeob

    2013-12-01

    Aphids are one of the most destructive pests in crop production such as pepper, cucumber, and eggplants. The importance of entomopathogenic fungi as alternative pest control agents is increasing. Conidia of entomopathogenic fungi are influenced by environmental conditions, such as temperature and relative humidity, and cause slow and fluctuating mortality. These factors have prevented wider application and use of biocontrol agents. For investigation of means of mitigation of such problems, we conducted bioassays with 47 fungal culture filtrates in order to evaluate the potential of secondary metabolites produced by entomopathogenic fungi for use in aphid control. Among 47 culture filtrates cultured potato dextrose broth, filtrate of Beauveria bassiana Bb08 showed the highest mortality (78%) against green peach aphid three days after treatments. Filtrate of Bb08 cultured in Adamek's medium showed higher toxicity as 100% to third instar nymphs of the aphid compared with seven other filtrates cultured in different broths amended with colloidal chitin or oil. The culture filtrates and fungal cultures from media amended with colloidal chitin or oil had lower control efficacies than filtrates without these additives in three different media. These results indicate that the fungal culture fluid or culture filtrate of B. bassiana Bb08 cultured in Adamek's medium has potential for development as a mycopesticide for aphid control.

  1. Beauveria bassiana Lipase A expressed in Komagataella (Pichia) pastoris with potential for biodiesel catalysis.

    Science.gov (United States)

    Vici, Ana C; da Cruz, Andrezza F; Facchini, Fernanda D A; de Carvalho, Caio C; Pereira, Marita G; Fonseca-Maldonado, Raquel; Ward, Richard J; Pessela, Benevides C; Fernandez-Lorente, Gloria; Torres, Fernando A G; Jorge, João A; Polizeli, Maria L T M

    2015-01-01

    Lipases (EC 3.1.1.3) comprise a biotechnologically important group of enzymes because they are able to catalyze both hydrolysis and synthesis reactions, depending on the amount of water in the system. One of the most interesting applications of lipase is in the biofuel industry for biodiesel production by oil and ethanol (or methanol) transesterification. Entomopathogenic fungi, which are potential source of lipases, are still poorly explored in biotechnological processes. The present work reports the heterologous expression and biochemical characterization of a novel Beauveria bassiana lipase with potential for biodiesel production. The His-tagged B. bassiana lipase A (BbLA) was produced in Komagataella pastoris in buffered methanol medium (BMM) induced with 1% methanol at 30°C. Purified BbLA was activated with 0.05% Triton X-100 and presented optimum activity at pH 6.0 and 50°C. N-glycosylation of the recombinant BbLA accounts for 31.5% of its molecular weight. Circular dichroism and molecular modeling confirmed a structure composed of α-helix and β-sheet, similar to α/β hydrolases. Immobilized BbLA was able to promote transesterification reactions in fish oil, demonstrating potential for biodiesel production. BbLA was successfully produced in K. pastoris and shows potential use for biodiesel production by the ethanolysis reaction.

  2. Beauveria bassiana Lipase A expressed in Komagataella (Pichia pastoris with potential for biodiesel catalysis

    Directory of Open Access Journals (Sweden)

    Ana Claudia Vici

    2015-10-01

    Full Text Available Lipases (EC 3.1.1.3 comprise a biotechnologically important group of enzymes because they are able to catalyze both hydrolysis and synthesis reactions, depending on the amount of water in the system. One of the most interesting application of lipase is in the biofuel industry for biodiesel production by oil and ethanol (or methanol transesterification. Entomopathogenic fungi, which are potential source of lipases, are still poorly explored in biotechnological processes. The present work reports the heterologous expression and biochemical characterization of a novel Beauveria bassiana lipase with potential for biodiesel production. The His-tagged B. bassiana lipase A (BbLA was produced in Komagataella pastoris in Buffered Methanol Medium (BMM induced with 1% methanol at 30 °C. Purified BbLA was activated with 0.05% Triton X-100 and presented optimum activity at pH 6.0 and 50°C. N-glycosylation of the recombinant BbLA accounts for 31.5% of its molecular weight. Circular dichroism and molecular modeling confirmed a structure composed of α-helix and β-sheet, similar to α/β hydrolases. Immobilized BbLA was able to promote transesterification reactions in fish oil, demonstrating potential for biodiesel production. BbLA was successfully produced in Komagataella pastoris and shows potential use for biodiesel production by the ethanolysis reaction.

  3. Analysis of whitefly transcriptional responses to Beauveria bassiana infection reveals new insights into insect-fungus interactions.

    Science.gov (United States)

    Xia, Jun; Zhang, Chang-Rong; Zhang, Shan; Li, Fang-Fang; Feng, Ming-Guang; Wang, Xiao-Wei; Liu, Shu-Sheng

    2013-01-01

    The fungal pathogen, Beauveria bassiana, is an efficient biocontrol agent against a variety of agricultural pests. A thorough understanding of the basic principles of insect-fungus interactions may enable the genetic modification of Beauveria bassiana to enhance its virulence. However, the molecular mechanism of insect response to Beauveria bassiana infection is poorly understood, let alone the identification of fungal virulent factors involved in pathogenesis. Here, next generation sequencing technology was applied to examine the expression of whitefly (Bemisia tabaci) genes in response to the infection of Beauveria bassiana. Results showed that, compared to control, 654 and 1,681genes were differentially expressed at 48 hours and 72 hours post-infected whiteflies, respectively. Functional and enrichment analyses indicated that the DNA damage stimulus response and drug metabolism were important anti-fungi strategies of the whitefly. Mitogen-activated protein kinase (MAPK) pathway was also likely involved in the whitefly defense responses. Furthermore, the notable suppression of general metabolism and ion transport genes observed in 72 hours post-infected B. tabaci might be manipulated by fungal secreted effectors. By mapping the sequencing tags to B. bassiana genome, we also identified a number of differentially expressed fungal genes between the early and late infection stages. These genes are generally associated with fungal cell wall synthesis and energy metabolism. The expression of fungal cell wall protein genes might play an important role in fungal pathogenesis and the dramatically up-regulated enzymes of carbon metabolism indicate the increasing usage of energy during the fungal infection. To our knowledge, this is the first report on the molecular mechanism of fungus-whitefly interactions. Our results provide a road map for future investigations on insect-pathogen interactions and genetically modifying the fungus to enhance its efficiency in whitefly

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

    Science.gov (United States)

    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 ...

  5. Potential use of the fungus Beauveria bassiana against the western flower thrips Frankliniella occidentalis without reducing the effectiveness of its natural predator Orius sauteri (Hemiptera: Anthocoridae)

    Science.gov (United States)

    Orius sauteri (Poppius) (Hemiptera: Anthocoridae) is an important predator of western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). Orius sauteri would be directly exposed to the entomopathogenic fungus Beauveria bassiana (Bals.) Vuillemin in the field should the fu...

  6. Evaluacion del efecto de Beauveria bassiana en el control biológico de Varroa destructor, parasito de la abeja melífera (Apis mellifera en la finca Felisa en el municipio de los Patios, Norte de Santander - Evaluacion del efecto de Beauveria bassiana en el control biológico de Varroa destructor, parasito de la abeja melífera (Apis mellifera en la finca Felisa en el municipio de los Patios, Norte de Santander

    Directory of Open Access Journals (Sweden)

    Francy Liliana Duarte

    2013-08-01

    Full Text Available Beekeeping is an activity that produces significant benefits to agriculture and the environment; Through the pollination action of bees helps to increase productivity as well as biodiversity in the ecosystem. In recent years, has significantly increased prevalence of parasites in this species, particularly the Varroasis, a disease caused by the mite Varroa destructor. The causal agent produces losses between 30% and 50% of total production and in turn can cause damage to the quality of honey by the excessive use of chemicals for disease control and the slow and progressive deterioration health of producers. Noting this, we evaluated the incidence of fungus Beauveria bassiana in natural populations of Varroa destructor in acarofauna associated, and the remains of the hive by pathogenicity tests in laboratory conditions to evaluate the effect of the biocontrol entomopathogenic then enter in apiaries infected with the disease. According to the results obtained in the adult infestation of V. destructor in Apis mellifera Africanized sampling Felisa made on the farm in the municipality of Los Patios, recorded rates of infestation in hives from 3.4% to 8.3% on the infection status of breeding was 4.5% to 13.7%. In laboratory tests it was possible to observe thatthe fungus B. bassiana attacked by an effective control mites without harming bees, propolis and honey allowing the insect to continue their normal activities, controlling the disease in a biological, not chemical.

  7. 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

  8. Interactions between the entomopathogenic fungi Beauveria bassiana (Ascomycota: Hypocreales) and the aphid parasitoid Diaeretiella rapae (Hymenoptera: Braconidae) on Myzus persicae (Hemiptera: Aphididae).

    Science.gov (United States)

    Silva, R J; Alencar, J R D C C; Silva, K P; Cividanes, F J; Duarte, R T; Agostini, L T; Polanczyk, R A

    2014-06-01

    The interactions between the entomopathogenic fungus Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Ascomycota: Hypocreales) and the aphid parasitoid Diaeretiella rapae McIntoch (Hymenoptera: Braconidae) were evaluated under laboratory conditions. Nymphs of Myzus persicae Sulzer (Hemiptera: Aphididae) were first exposed to parasitoid females for 24 h and then 0, 24, and 48 h afterwards sprayed with a solution of B. bassiana. Likewise, aphids were also sprayed with B. bassiana and then exposed to parasitoids at 0, 24, and 48 h afterwards. Parasitism rate varied from 13 to 66.5%, and were significantly lower in treatments where the two agents were exposed within a 0-24 h time interval compared with the control (without B. bassiana). Parasitoid emergence was negatively affected in treatments with B. bassiana spraying and subsequent exposure to D. rapae. Decreases in longevity of adult females of the D. rapae F1 generation were observed in treatments with B. bassiana spraying. The application of these two biological control agents can be used in combination on the control of M. persicae, wherein this use requires effective time management to avoid antagonistic interactions.

  9. A Distinctive Pattern of Beauveria bassiana-biotransformed Ginsenoside Products Triggers Mitochondria/FasL-mediated Apoptosis in Colon Cancer Cells.

    Science.gov (United States)

    Gum, Sang Il; Rahman, Md Khalilur; Won, Jong Soon; Cho, Min Kyung

    2016-01-01

    Ginseng is one of the most commonly used adaptogens. Transformation into the minor ginsenosides produces compounds with more effective action. Beauveria bassiana, a teleomorph of Cordyceps bassiana, is a highly efficient producer of mammalian steroids and produces large amounts of sugar-utilizing enzymes. However, the fermentation of steroid glycosides in ginseng with B. bassiana has never been studied. Thus, we evaluated the bioconversion of the major ginsenosides in white ginseng by B. bassiana. Interestingly, B. bassiana increased the total amount of protopanaxadiols and hydrolyzed Rb1 into minor ginsenosides, exhibiting high levels of Rd and Rg3, as well as moderate levels of Rb2 and Rc analyzed by high-performance liquid chromatography coupled with evaporative light-scattering detection. The β-glucosidase activity was highly increased, which led to the selective elimination of sugar moiety at the 20-C position of Rb1 to Rd, followed by Rg3. Rb2 and Rc accumulated because of the minimal activities of α-L-arabinopyranosidase and α-L-arabinofuranosidase, respectively. The fermentation product exerted dose-dependent cytotoxicity in HCT-15 cells, which are resistant to ginseng. The product, but not white ginseng, exhibited apoptotic effects via the Fas ligand and caspase 8/9. This study demonstrates for the first time that the B. bassiana-fermented metabolites have potent apoptotic activity in colon cancer cells, linking to a therapeutic use. Copyright © 2015 John Wiley & Sons, Ltd.

  10. Crescimento in vitro de isolados de Armillaria sp. obtidos de Pinus elliottii var: elliottii sob várias temperaturas In vitro growth of Armillaria sp. isolates obtained from Pinus elliottii var: elliottii under several temperatures

    Directory of Open Access Journals (Sweden)

    Nei Sebastião Braga Gomes

    2007-06-01

    Full Text Available A armilariose tem sido considerada a principal doença em Pinus no Brasil. Os sintomas e danos consistem no amarelecimento de acículas, declínio, podridão de raízes, exsudação de resina e morte. A temperatura é um dos fatores ambientais que influencia patógenos, doença de plantas ou ambos. Este trabalho avaliou o comportamento de três isolados de Armillaria sp. obtidos de P. elliottii var. elliottii, submetidos a uma faixa de temperatura de 16 a 26 ºC, utilizando a biomassa seca produzida em meio líquido como parâmetro de análise. Verificou-se que todos os isolados apresentaram máxima produção de biomassa a 22 ºC. Utilizando-se de regressão cúbica encontrou-se temperaturas de máximo crescimento entre 21,79 e 23,19 ºC. De acordo com os resultados, a melhor temperatura para crescimento dos isolados testados situou-se em 22 ºC.Armillaria root rot is the major pine disease in Brazil. Symptoms of this disease are yellowing of the needles, decline, root rot, resin exsudation and plant death. Temperature is an environmental factor that affects pathogens, the disease or both. This work evaluated the behaviour of three isolates of Armillaria sp. obtained from P. elliottii var. elliottii. The fungus isolates were submitted to temperature ranging from 16 to 26 ºC, by using dry biomass production in liquid medium as a measureable variable. All isolates produced higher amount of biomass at 22 ºC. A cubic regression showed a maximum point of temperatures between 21,8 e 23,2 ºC. The best temperature for fungus isolates growth was around 22 ºC.

  11. Polyphasic characterization of Gluconacetobacterdiazotrophicus isolates obtained from different sugarcane varieties Caracterização polifásica de isolados de Gluconacetobacterdiazotrophicus obtidos de diferentes variedades de cana-de-açúcar

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    Helma V. Guedes

    2008-12-01

    Full Text Available A polyphasic approach was applied to characterize 35 G. diazotrophicus isolates obtained from sugarcane varieties cultivated in Brazil. The isolates were analyzed by phenotypic (use of different carbon sources and genotypic tests (ARDRA and RISARFLP techniques. Variability among the isolates was observed in relation to the carbon source use preference. Glucose and sucrose were used by all isolates in contrast to myo-inositol, galactose and ribose that were not metabolized. The results of the analysis showed the presence of two groups clustered at 68% of similarity. The genetic distance was higher when RISA-RFLP analysis was used. Analysis of 16S rDNA sequences from isolates showed that all of them belonged to the G. diazotrophicus species. Neither effect of the plant part nor sugarcane variety was observed during the cluster analysis. The observed metabolic and genetic variability will be helpful during the strain selection studies for sugarcane inoculation in association with sugarcane breeding programs.Foi realizado a caracterização polifásica de 35 isolados obtidos de variedades de cana-de-açúcar cultivadas no Brasil, através de testes fenotípicos (uso de fontes diferentes de carbono e genotípicos (técnicas de ARDRA e RISA-RFLP. Houve variação entre os isolados com relação à utilização de fontes de carbono. Glicose e sacarose foram usadas por todos isolados, diferentemente de mio-inositol, galactose e ribose que não foram metabolizados. Os resultados da análise polifásica dos dados confirmam a formação de dois grupos, que apresentaram 68% de similaridade. Observou-se maior distância genética entre os isolados quando a técnica de RISA-RFLP foi aplicada. O sequênciamento da região 16S do rDNA mostrou que todos os isolados pertencem à espécie G. diazotrophicus. Não foi observado efeito da parte da planta ou variedade de cana-de-açúcar no agrupamento dos isolados. Em conjunto, esses resultados poderão auxiliar no

  12. Cordyceps bassiana inhibits smooth muscle cell proliferation via the ERK1/2 MAPK signaling pathway.

    Science.gov (United States)

    Jin, Enze; Han, Seongho; Son, Mina; Kim, Sung-Whan

    2016-01-01

    Cordyceps belongs to a genus of acormycete fungi and is known to exhibit various pharmacological effects. The aim of this study was to investigate the effect of Cordyceps species on the proliferation of vascular smooth muscle cells (VSMC) and their underlying molecular mechanism. A cell proliferation assay showed that Cordyceps bassiana ethanol extract (CBEE) significantly inhibited VSMC proliferation. In addition, neointimal formation was significantly reduced by treatment with CBEE in the carotid artery of balloon-injured rats. We also investigated the effects of CBEE on the extracellular signal-regulated kinase (ERK) signal pathway. Western blot analysis revealed increased ERK 1/2 phosphorylation in VSMCs treated with CBEE. Pretreatment with U0126 completely abrogated CBEE-induced ERK 1/2 phosphorylation. In conclusion, CBEE exhibited anti-proliferative properties that affected VSMCs through the ERK1/2 MAPK signaling pathway. Our data may elucidate the inhibitory mechanism of this natural product.

  13. Characterization of the Hog1 MAPK pathway in the entomopathogenic fungus Beauveria bassiana.

    Science.gov (United States)

    Liu, Jing; Wang, Zhi-Kang; Sun, Huan-Huan; Ying, Sheng-Hua; Feng, Ming-Guang

    2017-01-11

    High-osmolarity glycerol (HOG) pathway required for yeast osmoregulation relies upon the mitogen-activated protein kinase (MAPK) Hog1 cascade that comprise the MAPKKKs Ssk2/Ssk22 and Ste11 converging on the MAPKK Pbs2. Here we show a Hog1 cascade with the unique MAPKKK Ssk2 acting in Beauveria bassiana. Hypersensitivity to high osmolarity and high resistance to fludioxonil fungicide appeared in Δssk2, Δpbs2 and Δhog1 mutants whereas the two hallmark phenotypes were reversed in Δste11. Increased sensitivity to heat shock and decreased sensitivity to cell wall perturbation also occurred in the three mutants but not in Δste11 although antioxidant phenotypes were different in all deletion mutants. Intriguingly, signals of Hog1 phosphorylation induced by osmotic, oxidative and thermal cues were present in Δste11 but absent in Δssk2 and Δpbs2. Moreover, vegetative growth on minimal media with different carbon/nitrogen sources was much more suppressed in Δste11 and Δssk2 than in Δpbs2 and Δhog1 although all mutants suffered similar, but severe, conidiation defects on a standard medium. Normal host infection was abolished in Δste11 while virulence was differentially attenuated in other mutants. Our findings exclude Ste11 from the Hog1 cascade that regulates multiple stress responses and environmental adaptation of B. bassiana and perhaps other filamentous fungi. This article is protected by copyright. All rights reserved.

  14. Nanocomposite biofilms obtained from Whitemouth croaker (Micropogonias furnieri) protein isolate and Montmorillonite: evaluation of the physical, mechanical and barrier properties; Biofilmes nanocompositos obtidos de isolado proteico de corvina (Micropogonias furnieri) e Montmorilonita: avaliacao das propriedades fisicas, meanicas e de barreira

    Energy Technology Data Exchange (ETDEWEB)

    Cortez-Vega, William Renzo, E-mail: williamvega@ufgd.edu.br [Universidade Federal da Grande Dourados (UFGD), MS (Brazil). Fac. de Engenharia; Bagatini, Daniela Cardozo; Souza, Juliana Tais Andreghetto de; Prentice, Carlos, E-mail: danielabagatini@hotmail.com, E-mail: ju.andreghetto@hotmail.com, E-mail: dqmprent@furg.b [Universidade Federal do Rio Grande (FURG), RS (Brazil). Escola de Quimica e Alimentos

    2013-06-15

    The objective of this study was to evaluate the properties of nanocomposite biofilms based on Whitemouth croaker (Micropogonias furnieri) protein isolate with organophilic clays. Initially the croaker protein isolate (CPI) was obtained using the pH shifting process from by-products of croaker industrialization. A Box and Behnken experimental design was used to develop the films, with three levels of CPI (2, 3.5 and 5 g.100 g{sup -1} solution), montmorillonite MMT clay (0.3, 0.5 and 0.7 g.100 g{sup -1} solution) and glycerol (25, 30 and 35 g.100g{sup -1} CPI). The polymeric films were produced by the 'casting technique'. The tensile strength values ranged from 7.2 to 10.7 MPa and the elongation values from 39.6 to 45.8 %. The water vapor permeability (WVP) values ranged from 3.2 to 5.5 (g.mm.m{sup -2}.d{sup -1}) and the CPI had an average protein content of 97.87 % protein (d. b.). It was concluded that the nanocomposite films produced from CPI with MMT were promising from the standpoint of their mechanical properties, visual appearance and easy handling, as well as for their low water vapor permeability and low water solubility. With respect to their mechanical properties, the concentrations of CPI and MMT were the main factors influencing the development of the nanocomposite films. The results obtained from the experimental design indicated that 3.5 g of CPI.100 g{sup -1}solution, 0.5 g of MMT.100 g{sup -1} solution and 30 g of glycerol.100 g{sup -1} CPI would be the ideal parameters for the development of nanocomposite films by 'casting'. (author)

  15. Alteration of media composition and light conditions change morphology, metabolic profile, and beauvericin biosynthesis in Cordyceps bassiana mycelium.

    Science.gov (United States)

    Hyun, Sun-Hee; Lee, Seok-Young; Park, Shin Jung; Kim, Da Yeon; Chun, Young-Jin; Sung, Gi-Ho; Kim, Seong Hwan; Choi, Hyung-Kyoon

    2013-01-01

    Metabolic alterations of Cordyceps bassiana mycelium were investigated under the following culture medium and light conditions: dextrose agar supplemented with 0.5% yeast extract (SDAY) medium with light (SL), SDAY medium without light (SD), nut medium without light (ND), and iron-supplemented SDAY medium without light (FD). The levels of asparagine, aspartic acid, glutamic acid, glutamine, histidine, lysine, ornithine, and proline were significantly higher under SD and SL conditions. The levels of most of the alcohols, saturated fatty acids, unsaturated fatty acids, fatty acid esters, sterols, and terpenes were higher under the ND condition than in the other conditions, but beauvericin was not detectable under the ND condition. The FD condition was favorable for the enhanced production of aminomalonic acid, malic acid, mannonic acid, and erythritol. Thus, the metabolic characteristics of C. bassiana can be manipulated by varying the cultivation conditions, rendering this fungus potentially favorable as a nutraceutical and medicinal resource.

  16. 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%.

  17. Evaluation of eye irritation by S-(-)-10,11-dihydroxyfarnesic acid methyl ester secreted by Beauveria bassiana CS1029.

    Science.gov (United States)

    Son, Hyeong-U; Lee, Sang-Han

    2013-10-01

    The aim of this study was to investigate whether S-(-)-10,11-dihydroxyfarnesic acid methyl ester produced by cell subtype Beauveria bassiana CS1029 causes acute toxicity when used for cosmetic purposes by performing an eye irritation test. New Zealand white (NZW) rabbits were treated with a 100 mg/dose of S-(-)-10,11-dihydroxyfarnesic acid methyl ester according to standard procedure guidelines. No significant changes in terms of ocular lesions of the cornea, turbidity of the cornea, swelling of the eyelid or ocular discharge were observed in the methyl ester-treated groups, while sodium dioctyl sulfosuccinate, a positive control, caused severe toxicity. The anatomical and pathological observations indicate that the methyl ester produced by Beauveria bassiana CS1029 did not induce eye irritation in the lenses of the rabbits. The data suggest that the methyl ester evaluated in this study has promising potential as a cosmetic ingredient that does not irritate the eye.

  18. 不同球孢白僵菌菌株生物学特性与其对家蚕致病力的关系%Relationship between Biological Characteristics of Beauveria bassiana ( Bals.) Vuill and Pathogenicity to Bombyx mori L.

    Institute of Scientific and Technical Information of China (English)

    骆海玉; 邓业成; 廖永梅; 李瑞钰

    2012-01-01

    [目的]探讨球孢白僵菌菌株的生物学特性与其对家蚕致病力的关系,为家蚕白僵病的防治提供科学依据.[方法]从我国各地收集6个白僵菌杀虫剂,并从广西养蚕区采集3个僵蚕样品,对从中分离纯化的菌株进行形态观察和分子鉴定,同时对家蚕进行致病力测定,比较各菌株菌落直径、产孢量、胞外蛋白酶产生水平等生物学特征指标的差异.[结果]获得的9个分离菌株均为球孢白僵菌(Beauveria bassiana Vuillemin);各菌株对家蚕都有较强致病力,且致病力有一定差异;各菌株菌落生长直径、产孢量及胞外蛋白酶活性均存在较大差异,并与其对家蚕的致病力之间存在相关关系.[结论]各菌株产孢量和胞外蛋白酶活性与其对家蚕的致病力之间存在显著正相关.%[Objective] This study was to investigate the relationship between biological characteristics of Beauveria bassiana ( Bals. ) Yuill and pathogenicity of Bombyx mori L. , with the aim to provide scientific basis for the control of white muscardine in Bombyx mori L. . [ Method ] The strains isolated and purified from the 6 Beauveria bassiana biocontrol agents from all over the country and the 3 white muscardine silkworms collected from Guangxi provincial silkworm rearing areas were identified by the morphological observation and molecular biology technology. The pathogenicity of silkworms to B. bassaina was determined, and the biological characteristics such as growth diameter, spore production amount and the extracellular protease activity of the different B. bassiana strains were compared. [ Result] The isolated 9 strains were al] B. bassaina (Bals. ) Vuillemin, and all strains had high pathogenicity to silkworm, but with different pathogenicities. The growth diameter, spore production amount and extracellular protease activity of different B. bassiana strains were also different, and showed correlation with the patheogenicity to silkworms

  19. The entomopathogenic fungal endophytes Purpureocillium lilacinum (formerly Paecilomyces lilacinus) and Beauveria bassiana negatively affect cotton aphid reproduction under both greenhouse and field conditions.

    Science.gov (United States)

    Castillo Lopez, Diana; Zhu-Salzman, Keyan; Ek-Ramos, Maria Julissa; Sword, Gregory A

    2014-01-01

    The effects of two entomopathogenic fungal endophytes, Beauveria bassiana and Purpureocillium lilacinum (formerly Paecilomyces lilacinus), were assessed on the reproduction of cotton aphid, Aphis gossypii Glover (Homoptera:Aphididae), through in planta feeding trials. In replicate greenhouse and field trials, cotton plants (Gossypium hirsutum) were inoculated as seed treatments with two concentrations of B. bassiana or P. lilacinum conidia. Positive colonization of cotton by the endophytes was confirmed through potato dextrose agar (PDA) media plating and PCR analysis. Inoculation and colonization of cotton by either B. bassiana or P. lilacinum negatively affected aphid reproduction over periods of seven and 14 days in a series of greenhouse trials. Field trials were conducted in the summers of 2012 and 2013 in which cotton plants inoculated as seed treatments with B. bassiana and P. lilacinum were exposed to cotton aphids for 14 days. There was a significant overall effect of endophyte treatment on the number of cotton aphids per plant. Plants inoculated with B. bassiana had significantly lower numbers of aphids across both years. The number of aphids on plants inoculated with P. lilacinum exhibited a similar, but non-significant, reduction in numbers relative to control plants. We also tested the pathogenicity of both P. lilacinum and B. bassiana strains used in the experiments against cotton aphids in a survival experiment where 60% and 57% of treated aphids, respectively, died from infection over seven days versus 10% mortality among control insects. Our results demonstrate (i) the successful establishment of P. lilacinum and B. bassiana as endophytes in cotton via seed inoculation, (ii) subsequent negative effects of the presence of both target endophytes on cotton aphid reproduction using whole plant assays, and (iii) that the P. lilacinum strain used is both endophytic and pathogenic to cotton aphids. Our results illustrate the potential of using these

  20. The entomopathogenic fungal endophytes Purpureocillium lilacinum (formerly Paecilomyces lilacinus and Beauveria bassiana negatively affect cotton aphid reproduction under both greenhouse and field conditions.

    Directory of Open Access Journals (Sweden)

    Diana Castillo Lopez

    Full Text Available The effects of two entomopathogenic fungal endophytes, Beauveria bassiana and Purpureocillium lilacinum (formerly Paecilomyces lilacinus, were assessed on the reproduction of cotton aphid, Aphis gossypii Glover (Homoptera:Aphididae, through in planta feeding trials. In replicate greenhouse and field trials, cotton plants (Gossypium hirsutum were inoculated as seed treatments with two concentrations of B. bassiana or P. lilacinum conidia. Positive colonization of cotton by the endophytes was confirmed through potato dextrose agar (PDA media plating and PCR analysis. Inoculation and colonization of cotton by either B. bassiana or P. lilacinum negatively affected aphid reproduction over periods of seven and 14 days in a series of greenhouse trials. Field trials were conducted in the summers of 2012 and 2013 in which cotton plants inoculated as seed treatments with B. bassiana and P. lilacinum were exposed to cotton aphids for 14 days. There was a significant overall effect of endophyte treatment on the number of cotton aphids per plant. Plants inoculated with B. bassiana had significantly lower numbers of aphids across both years. The number of aphids on plants inoculated with P. lilacinum exhibited a similar, but non-significant, reduction in numbers relative to control plants. We also tested the pathogenicity of both P. lilacinum and B. bassiana strains used in the experiments against cotton aphids in a survival experiment where 60% and 57% of treated aphids, respectively, died from infection over seven days versus 10% mortality among control insects. Our results demonstrate (i the successful establishment of P. lilacinum and B. bassiana as endophytes in cotton via seed inoculation, (ii subsequent negative effects of the presence of both target endophytes on cotton aphid reproduction using whole plant assays, and (iii that the P. lilacinum strain used is both endophytic and pathogenic to cotton aphids. Our results illustrate the potential of

  1. The effect of Mirabilis jalapa leaves biopesticide treatment on the mycelium growth of entomopathogenic fungi Beauveria bassiana inside the larvae body Crocidolomia binotalis

    Science.gov (United States)

    Pramita, Mia; Anggraeni, Tjandra

    2015-09-01

    Pest control with biological method (biopesticide and entomopathogenic fungi) is an alternative program to reduce application of chemical insecticide. Biopesticide of Mirabilis jalapa leaves has been discovered rich in secondary metabolites which has antifeedant activity that can provide physiological interference in insect larvae and the generation numbers[1]. Entomopathogenic fungi Beauveria bassiana has potential to control pest populations[2]. The growth of mycelium B. bassiana may interfere metabolism process inside the host body. Otherwise, B. bassiana produce toxins such as beauvericin that can increase mortality of pest. Combination of M. jalapa and B. bassiana reduce LT50 on C. binotalis larvae[3]. Thus, this study aims to determine influence of provision of biopesticide M. jalapa leaves on growth of mycelium entomopathogenic fungi B. bassiana inside larvae body C. binotalis and to detect the presence of beauvericin in vivo. Third instar larvae of C. binotalis were divided into a control, fungal and combination group. The combination group was given biopesticide and fungi. The concentration of biopesticide was 0.8% (w/v) and concentration of fungi spores was 107 spores/ml. Spores (vol. 5µl) done topically to larvae in interval 6 hours after treatment of biopesticide on non-pesticide cabbage leaves. Afterwards, histological observations performed at 24, 48, 72, 96 hours after treatment. The result show of emergence hyphae and mycelium growth inside lumen of larvae midgut on combination group faster than fungal group. This is thought to be caused by the influence of secondary metabolites of biopesticide M. jalapa leaves. In addition, beauviricin is detectable both of fungal and combination group. Thus, it can be concluded that treatment of biopesticide from M. jalapa leaves can accelerate on growth of mycelium entomopathogenic fungi B. bassiana inside the larvae body C. binotalis and toxic of B. bassiana such as beauvericin was detected on fungal and

  2. Microbiological study of naturally fermented Algerian green olives: isolation and identification of lactic acid bacteria and yeasts along with the effects of brine solutions obtained at the end of olive fermentation on Lactobacillus plantarum...

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    Nour-Eddine, Karam

    2006-09-01

    Full Text Available The microflora of naturally fermented green olives produced in Western Algeria was studied over 15, 60 and 90 day fermentation periods. Different microorganisms (aerobic bacteria, coliforms, staphylococci, lactic acid bacteria, lactobacilli, enterococci, yeasts, psychrotrophs and lipolytic bacteria were recorded at 15 and 60 days of fermentation. After 90 days (pH 4.40 of fermentation, the lactic acid bacteria population became dominant and persisted together with yeasts throughout the fermentation period. The lactic acid bacteria isolated (343 isolates were identified as L. casei, L. rhamnosus, L. paracasei, L. plantarum, L. lactis subsp. lactis, E. faecalis, E. faecium and E. durans. The dominant species was L. plantarum. Yeasts were isolated from all samples (32 isolates and were identified as Saccharomyces cerevisiae or Candida parapsilosis. Also, in this study we reported that brine solutions obtained at the end of olive fermentation were able to stimulate the growth of several L. plantarum strainsLa microflora de las aceitunas verdes fermentadas naturalmente elaboradas en Argelia Occidental fue estudiada en períodos de fermentación de 15, 60 y 90 días. Diferentes microorganismos (bacterias aeróbicas, coliformes, estafilococos, bacterias del ácido láctico, lactobacilos, enterococos, levaduras, psicotrofos y bacterias lipolíticas fueron detectados a los 15 y 60 días de fermentación. Después de 90 días de fermentación (pH 4.40, la población de bacterias lácticas se hizo dominante y persistió junto con las levaduras a lo largo de todo el proceso. Las bacterias lácticas aisladas (343 fueron identificadas como L. casei, L. rhamnosus, L. paracasei, L. plantarum, L. lactis subsp. lactis, E. faecalis, E. faecium y E. durans. La especie dominante fue L. plantarum. Las levaduras aisladas (32 de todas las muestras fueron identificadas como Saccharomyces cerevisiae o Candida parapsilosis. También se recoge en este estudio que las

  3. Metabolic responses of Beauveria bassiana to hydrogen peroxide-induced oxidative stress using an LC-MS-based metabolomics approach.

    Science.gov (United States)

    Zhang, Chen; Wang, Wei; Lu, Ruili; Jin, Song; Chen, Yihui; Fan, Meizhen; Huang, Bo; Li, Zengzhi; Hu, Fenglin

    2016-06-01

    The entomopathogenic fungus, Beauveria bassiana, is commonly used as a biological agent for pest control. Environmental and biological factors expose the fungus to oxidative stress; as a result, B. bassiana has adopted a number of anti-oxidant mechanisms. In this study, we investigated metabolites of B. bassiana that are formed in response to oxidative stress from hydrogen peroxide (H2O2) by using a liquid chromatography mass spectrometry (LC-MS) approach. Partial least-squares discriminant analysis (PLS-DA) revealed differences between the control and the H2O2-treated groups. Hierarchical cluster analysis (HCA) showed 18 up-regulated metabolites and 25 down-regulated metabolites in the H2O2-treated fungus. Pathway analysis indicated that B. bassiana may be able to alleviate oxidative stress by enhancing lipid catabolism and glycometabolism, thus decreasing membrane polarity and preventing polar H2O2 or ROS from permeating into fungal cells and protecting cells against oxidative injury. Meanwhile, most of the unsaturated fatty acids that are derived from glycerophospholipids hydrolysis can convert into oxylipins through autoxidation, which can prevent the reactive oxygen of H2O2 from attacking important macromolecules of the fungus. Results showed also that H2O2 treatment can enhance mycotoxins production which implies that oxidative stress may be able to increase the virulence of the fungus. In comparison to the control group, citric acid and UDP-N-acetylglucosamine were down-regulated, which suggested that metabolic flux was occurring to the TCA cycle and enhancing carbohydrate metabolism. The findings from this study will contribute to the understanding of how the molecular mechanisms of fungus respond to environmental and biological stress factors as well as how the manipulation of such metabolisms may lead to selection of more effective fungal strains for pest control.

  4. The role of fungus Beauveria bassiana in reducing the number of Pissodes castaneus (Col., Curculionidae in young forests

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    Skrzecz Iwona

    2016-12-01

    Full Text Available Small banded pine weevil Pissodes castaneus is one of the most dangerous pests of Pinus sylvestris plantations and thickets. The lack of effective and environmentally safe methods of limiting the number of the pest justified to undertake the studies aimed at the laboratory and field evaluation of biological activity of entomopathogenic fungus Beauveria bassiana used to reduce the numbers of small banded pine weevil.

  5. 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.

  6. PENAMBAHAN BEBERAPA JENIS BAHAN NUTRISI PADA MEDIA PERBANYAKAN UNTUK MENINGKATKAN VIRULENSI BEAUVERIA BASSIANA TERHADAP HAMA WALANG SANGIT

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    Ni Siluh Putu Nuryanti

    2013-09-01

    Full Text Available A research was conducted to increase the virulence of Beauvaria bassiana to the rice bug, L.  acuta, by adding four different substances (materials to the rice media to grow the fungus. The research was conducted from April to August 2011 at Polinela Laboratory, Bandar Lampung.  To do this a set of experimental unit consisting of five treatments and five replications were arranged in completely randomized design. The  four treatments were media made up of rice (rice based media each given or added with four different materials such as rice skin powder, corn sugar, grasshopper powder and chitosan powder. The result showed that B. bassiana grown on rice media added with grasshopper powder or rice skin powder were able of causing mortality to the L. acuta as high as 78% and 71%, respectively, meaning that both mortalities were much higher than that of other replication. The virulence values of Beauveria bassiana  grown on those both powder were also higher than that of three others.

  7. A novel bioassay to evaluate the potential of Beauveria bassiana strain NI8 and the Insect growth regulator novaluron against Lygus lineolaris on a non-autoclaved solid artificial diet

    Science.gov (United States)

    The entomopathogenic fungus Beauveria bassiana and the insect growth regulator novaluron are potential candidates for the control of Lygus lineolaris. Efforts are being made for their use in the Mississippi Delta. B. bassiana and novaluron highly affected TPB survival when they were applied directly...

  8. LABORATORY AND FIELD EVALUATION OF A CYPERMETHRIN-BASED INSECTICIDE FOR THE CONTROL OF ALPHITOBIUS DIAPERINUS PANZER (COLEOPTERA: TENEBRIONIDAE AND ITS IN-VITRO EFFECTS ON BEAUVERIA BASSIANA BALS. VUILL. (HYPOCREALES: CORDYCIPITACEAE

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    DGP Oliveira

    Full Text Available ABSTRACT The control of Alphitobius diaperinus is based mainly on chemical insecticides, and the indiscriminate and incorrect use of these products has led to failures in insect control. Thus, it is important to monitor the efficiency of the products available on the market and to develop alternative insect control strategies. The present study evaluated the effect of a new product (cypermethrin-6% under laboratory and field conditions and assessed its in-vitro compatibility with the fungus Beauveria bassiana (Unioeste 4 isolate. Its efficiency in dry powder and wettable powder formulations was also assessed in the laboratory through comparison with a similar insecticide (cypermethrin-5% at five different concentrations. The field assays were conducted in two commercial broiler breeder houses (Treated and Control, applying the new product on the litter at the manufacturer's recommended concentration (RC. Germination, vegetative growth, conidial production, number of colony forming units (CFUs and insecticidal activity of the fungus were used to evaluate the compatibility of the insecticide. The effects of the cypermethrin-6% product and cypermethrin-5% were equivalent in the laboratory, and the new product exhibited better performance at lower concentrations (1/4RC, ⅛RC. The strategy applied in the field reduced the insect population in up to 96% after 75 days. Additionally, all concentrations of cypermethrin-6% were compatible with the fungus under the evaluated in-vitro conditions. Therefore, the new product is considered selective for B. bassiana. Further studies are necessary to assess its compatibility under field conditions, consolidating this strategy as a viable alternative for managing this pest.

  9. Production of high quality brain-derived neurotrophic factor (BDNF) and tropomyosin receptor kinase B (TrkB) RNA from isolated populations of rat spinal cord motor neurons obtained by Laser Capture Microdissection (LCM).

    Science.gov (United States)

    Mehta, Prachi; Premkumar, Brian; Morris, Renée

    2016-08-03

    The mammalian central nervous system (CNS) is composed of multiple cellular elements, making it challenging to segregate one particular cell type to study their gene expression profile. For instance, as motor neurons represent only 5-10% of the total cell population of the spinal cord, meaningful transcriptional analysis on these neurons is almost impossible to achieve from homogenized spinal cord tissue. A major challenge faced by scientists is to obtain good quality RNA from small amounts of starting material. In this paper, we used Laser Capture Microdissection (LCM) techniques to identify and isolate spinal cord motor neurons. The present analysis revealed that perfusion with paraformaldehyde (PFA) does not alter RNA quality. RNA integrity numbers (RINs) of tissue samples from rubrospinal tract (RST)-transected, intact spinal cord or from whole spinal cord homogenate were all above 8, which indicates intact, high-quality RNA. Levels of mRNA for brain-derived neurotrophic factor (BDNF) or for its tropomyosin receptor kinase B (TrkB) were not affected by rubrospinal tract (RST) transection, a surgical procedure that deprive motor neurons from one of their main supraspinal input. The isolation of pure populations of neurons with LCM techniques allows for robust transcriptional characterization that cannot be achieved with spinal cord homogenates. Such preparations of pure population of motor neurons will provide valuable tools to advance our understanding of the molecular mechanisms underlying spinal cord injury and neuromuscular diseases. In the near future, LCM techniques might be instrumental to the success of gene therapy for these debilitating conditions.

  10. Characterisation of drug resistance of nosocomial ESBL-producing E. coli isolates obtained from a Turkish university hospital between 2009 and 2012 by pulsed field gel electrophoresis and antibiotic resistance tests.

    Science.gov (United States)

    Karagöz, Alper; Sunnetcioglu, Mahmut; Ceylan, Mehmet Resat; Bayram, Yasemin; Yalcin, Gozde; Kocak, Nadir; Suvak, Burak; Andac, Cenk A

    2016-01-01

    In this study, drug resistance of 28 ESBL-producing Escherichia coli isolates obtained from 144 patients hospitalized at the Yüzüncüyil University Hospital at Van (YUH), Turkey, between 2009 and 2012 were characterized by pulsed field gel electrophoresis and antibiotic susceptibility tests. Antibiotic resistance profile was determined by Phoenix automated system (BD, USA). The ratio of ESBL-producing E. coli strains was determined to be 19.4% (28 out of 144 E. coli isolates). It was determined that the anaesthesiology, paediatrics and thoracic medicine intensive care units in YUH were cross-contaminated between 2009 and 2012 by ESBL-producing E. coli strains, which is a sign of nosocomial infection in YUH. Analysis of PFGE results gave rise to two main PFGE profiles, profile-A with four subprofiles and profile-B with three subprofiles, where profile-A predominates over profile-B (14%). Comparison of the antibiotic resistance profile with the PFGE profile yielded similarities while some differences also exist due to either identical restriction enzyme cutting sites with slightly different genetic sequences in between the cutting sites or newly formed restriction enzyme cutting sites that do not affect antibiotic resistance genes. Enterobacteriaceae, particularly E. coli, have developed resistance in YUH by producing ESBLs against oxyimino and non-oxyimino cephalosporins, and penicillin-type antibiotics. Therefore, more effective antibiotics such as cefoxitin or cefoperazone-sulbactam should be used for the treatment of future nosocomial infections in YUH while hospital staff should take care with hygiene, such as hand washing.

  11. A method of multiplex PCR for detection of field released Beauveria bassiana, a fungal entomopathogen applied for pest management in jute (Corchorus olitorius).

    Science.gov (United States)

    Biswas, Chinmay; Dey, Piyali; Gotyal, B S; Satpathy, Subrata

    2015-04-01

    The fungal entomopathogen Beauveria bassiana is a promising biocontrol agent for many pests. Some B. bassiana strains have been found effective against jute pests. To monitor the survival of field released B. bassiana a rapid and efficient detection technique is essential. Conventional methods such as plating method or direct culture method which are based on cultivation on selective media followed by microscopy are time consuming and not so sensitive. PCR based methods are rapid, sensitive and reliable. A single primer PCR may fail to amplify some of the strains. However, multiplex PCR increases the possibility of detection as it uses multiple primers. Therefore, in the present investigation a multiplex PCR protocol was developed by multiplexing three primers SCA 14, SCA 15 and SCB 9 to detect field released B. bassiana strains from soil as well as foliage of jute field. Using our multiplex PCR protocol all the five B. bassiana strains could be detected from soil and three strains viz., ITCC 6063, ITCC 4563 and ITCC 4796 could be detected even from the crop foliage after 45 days of spray.

  12. 10株白僵菌菌株形态学与分子鉴定%Morphological and Molecular Identification of 10 Beauveria bassiana Strains

    Institute of Scientific and Technical Information of China (English)

    刘宝生; 谷希树; 田涛; 霍建飞; 孙淑琴

    2012-01-01

    对10株白僵菌属菌株的菌落形态、产孢结构和分生孢子形状、大小作了观察与测量,形态学鉴定为球孢白僵菌.同时对此10株球孢白僵菌核糖体DNA - ITS进行了测序分析,与NCBI数据库中的球孢白僵菌(Beauveria bassiana)多个菌株的同源性达99.10% ~99.64%.依此,10株白僵菌菌株分子鉴定结果为球孢白僵菌(Beauveria bassiana).%In this study, the colony morphology, spore structure and spore shape and size of 10 Beauveria strains were observed and measured, and they were initially identified as Beauveria bassiana. At the same time, their ribosomal DNA - ITS were sequenced and compared with that of Beauveria bassiana in NCBI database. The results showed the homology was 99.10% -99.64%. Eventually these 10 strains were identified as Beauveria bassiana.

  13. 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

  14. 响应面法优化球孢白僵菌CXJ-1的发酵工艺条件%Optimizatiom of Fermentation Conditons for Beauveria bassiana CXJ-1 Spore by Response Surface Methodology

    Institute of Scientific and Technical Information of China (English)

    孙佰平; 任娟; 赵思峰; 侯彩霞; 王斌; 刘启鹏

    2011-01-01

    Response surface methodology was used to optimize fermentation conditons in order to improve spore production of Beauveria bassiana CXJ-1. The optimum fermentation temperature, rotation speed and pH were obtained with one-factor-at-time. Through Plackett-Burman experimental design and Box-Behnken experimental design for response surface optimization, the optimal conditions for spore production of Beauveria bassiana CXJ-1 were also determined as follows: temperature 28.07℃ , rotation speed 198. 95 r/min and the pH value 6. 10. Under these conditions, the predicted value of the concentration of spores was 9.5X108 spores/mL and the verified value was 9. 7 X 108 spores/mL, the fermentation validation showed that the experimental value have high inosculating degree to model prediction.%为了提高球孢白僵菌CXJ-1(Beauveria bassiana CXJ-1)在液体发酵过程中分生孢子的产孢量,采用响应面法对球孢白僵菌CXJ-1产孢的发酵工艺条件进行优化.采用单因素实验确定了产孢发酵条件最适初始pH、发酵转速和发酵温度.通过Plackett-Burman实验设计和Box-Behnken实验设计响应面分析优化,确定了球孢白僵菌CXJ-1产孢最佳发酵工艺参数为:温度28.07℃,转速198.95 r/min,初始pH 6.10,预测孢子浓度可达9.5×108个/mL,在优化发酵条件下进行验证实验,孢子浓度达到9.7×108个/mL,表明验证值和预测值有较高的吻合度.

  15. JAK/STAT signaling pathway-mediated immune response in silkworm (Bombyx mori) challenged by Beauveria bassiana.

    Science.gov (United States)

    Geng, Tao; Lv, Ding-Ding; Huang, Yu-Xia; Hou, Cheng-Xiang; Qin, Guang-Xing; Guo, Xi-Jie

    2016-12-20

    Innate immunity was critical in insects defensive system and able to be induced by Janus kinase/signal transducer and activator of transcription cascade transduction (JAK/STAT) signaling pathway. Currently, it had been identified many JAK/STAT signaling pathway-related genes in silkworm, but little function was known on insect innate immunity. To explore the roles of JAK/STAT pathway in antifungal immune response in silkworm (Bombyx mori) against Beauveria bassiana infection, the expression patterns of B. mori C-type lectin 5 (BmCTL5) and genes encoding 6 components of JAK/STAT signaling pathway in silkworm challenged by B. bassiana were analyzed using quantitative real time PCR. Meanwhile the activation of JAK/STAT signaling pathway by various pathogenic micro-organisms and the affect of JAK/STAT signaling pathway inhibitors on antifungal activity in silkworm hemolymph was also detected. Moreover, RNAi assay of BmCTL5 and the affect on expression levels of signaling factors were also analyzed. We found that JAK/STAT pathway could be obviously activated in silkworm challenged with B. bassiana and had no response to bacteria and B. mori cytoplasmic polyhedrosis virus (BmCPV). However, the temporal expression patterns of JAK/STAT signaling pathway related genes were significantly different. B. mori downstream receptor kinase (BmDRK) might be a positive regulator of JAK/STAT signaling pathway in silkworm against B. bassiana infection. Moreover, antifungal activity assay showed that the suppression of JAK/STAT signaling pathway by inhibitors could significantly inhibit the antifungal activity in hemolymph and resulted in increased sensitivity of silkworm to B. bassiana infection, indicating that JAK/STAT signaling pathway might be involved in the synthesis and secretion of antifungal substances. The results of RNAi assays suggested that BmCTL5 might be one pattern recognition receptors for JAK/STAT signaling pathway in silkworm. These findings yield insights for better

  16. Protocols for obtainment and isolation of two mesenchymal stem cell sources in sheep Protocolos para obtenção e isolamento de células tronco mesenquimais de duas fontes em ovinos

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    Leandro Fadel

    2011-08-01

    Full Text Available PURPOSE: To evaluate different protocols to isolate stem cells from ovine umbilical cord blood and adipose tissue. METHODS: There were used 5 samples of umbilical blood and 5 samples of perirenal adipose tissue from 10 female sheep. All the samples were obtained through surgery, to harvest aseptic samples. There were used 3 protocols for obtainment and culture of umbilical cord blood stem cells and 4 protocols for ovine adipose tissue stem cells. RESULTS: It was possible to observe only one successful protocol for the obtainment of umbilical cord blood stem cells. When analyzing the techniques used to obtain adipose tissue stem cells, only one of the methods was effective as well. Through colony forming unit assay, there were obtained 58 colonies of cells after seven days in culture. Flow citometry tests revealed the cells were positive to CD44 and exhibited negative reaction to CD38, CD45, CD41/61. These cells showed a growth curve with very well defined phases LOG, LAG and PLATEAU. This phases are typically seem in mesenchymal stem cells growth curves. CONCLUSIONS: The isolation and culture of mesenchymal stem cells from ovine umbilical cord blood are complex and request more detailed assays. Stem cells from fat tissue sheep showed mesenchymal characteristics, according to their cell growth curve, ability to origin colonies of fibroblastoid cells and positive reactivity with the antibody CD44 by flow citometry.OBJETIVO: Testar diferentes protocolos para o isolamento de células tronco a partir de sangue de cordão umbilical e tecido adiposo de ovinos. MÉTODOS: Foram utilizadas cinco amostras de sangue de cordão umbilical e cinco amostras de tecido adiposo perirrenal de 10 fêmeas de ovelha. A coleta das amostras foi realizada através de procedimento cirúrgico para coleta do material de forma mais asséptica possível. Foram realizados três protocolos de isolamento e cultivo das células-tronco do cordão umbilical e quatro protocolos para

  17. Isolation, morphological identification and pathogenicity of Cylindrocladium scoparium and C. clavatum isolates obtained from plants rhizosphere cultivated in Pernambuco State Isolamento, identificação morfológica e patogenicidade de isolados de Cylindrocladium scoparium e C. clavatum da rizosfera de plantas cultivadas no Estado de Pernambuco

    Directory of Open Access Journals (Sweden)

    Vitorina Nerivânia Covello Rehn

    2004-12-01

    Full Text Available Twelve isolates of Cylindrocladium scoparium and 4 isolates of C. clavatum were obtained from the rhizosphere of various species of plants by baiting with Ricinus communis leaves. The isolates of C. scoparium developed conidia of 32-(45-60 x 3 -(4-5 µm, and pyriform to ellipsoidal vesicles. C. clavatum showed conidia of 36-(44-49 x 2-(4-6 µm and clavate vesicles. All isolates induced necrosis on leaves and hypocotyls of eucalypt seedlings, with varying expression of symptoms.Doze isolados de Cylindrocladium scoparium e 4 isolados de C. clavatum foram obtidos da rizosfera de diversas plantas usando folhas de Ricinus communis como isca. A primeira espécie mostrou conídios com dimensões de 32-(45-60 x 3 -(4-5 µm e vesícula piriforme a elipsoidal. Os conídios de C. clavatum apresentaram dimensões entre 36-(44-49 x 2-(4-6 µm e vesícula clavada. Em plântulas de eucalipto todos os isolados induziram necrose em folhas e hipocótilos, variando apenas na expressão dos sintomas.

  18. Changes in the Contents of Proteins and Amino Acids of Apriona germari Larvae Infected by Beauveria bassiana%桑天牛幼虫感染球孢白僵菌后蛋白质和氨基酸含量的变化

    Institute of Scientific and Technical Information of China (English)

    李会平; 黄大庄; 唐秀光

    2012-01-01

    To understand the relationship between the contents of proteins and free amino acids and the infection of Beauveria bassiana. Changes in the contents of proteins and free amino acids in haemolymph of Apriona germari larvae infected by B. bassiana were detected. The result showed that the content of proteins increased to the maximum in 3 days after inoculation,indicating the proteins and enzymes could be produced for the immunologic responses,and then declined. By using the polyacrylamide gelelectrophoresis( PAGE) technique, the change in protein composition in Apriona germari larvae infected by Beauveria bassiana was analyzed. The result showed that the band number increased after inoculation. There were significant differences in the band widths and staining intensities. The results of amino acids analysis showed that there were abundant necessary amino acids for the strain' s growth in A. germari larvae bodies, thus B. bassiana showed a high virulence to A. germari larvae. The content of free amino acids decreased in 24 h after inoculation, suggesting that the B. bassiana invaded into the insect body absorbed some amino acids from the blood as nutrition. The content of free amino acids had no further changes from 24 -72 h,and then increased sharply in 96 h and 120 h probably because of gradual loss of immunoregulation, destroy of tissue and disintegration of proteins. Before dead of the infected A. germari larvae,the contents of different amino acids changed differentially,suggesting that different growth phases of the isolate needed different amino acids.%桑天牛现称桑粒肩天牛(Apriona germari),是多种林木、果树、花卉的重要蛀干害虫,也是我国危害最严重、最难防治的害虫之一.近年来,其猖獗危害已成为我国养蚕业、林业及果树种植业等经济发展的严重障碍.球孢白僵菌(Beauveria bassiana)是一类广谱性昆虫病原真菌,能侵染15目149科的700多种昆虫(St Leger,2001;Blake et al.,2002

  19. Essential oils and Beauveria bassiana against Dermanyssus gallinae (Acari: Dermanyssidae): Towards new natural acaricides.

    Science.gov (United States)

    Immediato, Davide; Figueredo, Luciana Aguiar; Iatta, Roberta; Camarda, Antonio; de Luna, Rafaela Lira Nogueira; Giangaspero, Annunziata; Brandão-Filho, Sinval Pinto; Otranto, Domenico; Cafarchia, Claudia

    2016-10-15

    Essential oils (EOs) and entomopathogenic fungi such as Beauveria bassiana (Bb) strains have the potential to be used as alternative insecticides and acaricides for controlling ectoparasites as Dermanyssus gallinae. These compounds have some limitations in their use: the acaricidal effect of EOs is rapid, but short-lived, whilst that of Bb is delayed, but long-lived. To evaluate the effect of both compounds combined against D. gallinae, the non-toxic dose of Eucalyptus globulus, Eucalyptus citriodora, Thymus vulgaris and Eugenia caryophyllata essential oils were firstly calculated for "native" strains of Bb. Subsequently, the effects of the combination of selected EOs with Bb against nymph and adult poultry red mites (PRMs) was assessed. EO concentrations ranging from 0.0015 to 8% v/v (i.e., nine double dilutions) were used to evaluate their effect on germination, sporulation and vegetative growth rates of native strains of Bb. A total of 1440 mites (720 nymphs and 720 adults) were divided into three-treated group (TGs) and one control group (CG). In TGs, mites were exposed to Bb in combination with the selected EO (TG1), EO alone (TG2) or Bb (TG3) alone. In the CG, mites were exposed to 0.1% tween 80 plus EO solvent (CG). E. globulus and E. citriodora were toxic for Bb in concentrations higher than 0.2% and 0.003% respectively, whilst E. caryophyllata and T. vulgaris were toxic at all concentrations tested against Bb. Based on the results of the toxicity assays against Bb, E. globulus was chosen to be tested as acaricide resulting non-toxic for Bb at concentration lower than 0.4%. Increased mortality of D. gallinae adults was recorded in TG1 than those in other TGs from 4days post-infection (T+4DPI). A 100% mortality of D. gallinae was recorded in adults at T+9DPI and at T+10DPI in nymphs in TG1 and later than T+11DPI in the other TGs. Used in combination with E. globulus, Bb displayed an earlier acaricidal effect towards both haematophagous D. gallinae stages

  20. 分离自我国蜜蜂的一株新的致病螺原体及其基本特性%A new pathogenic spiroplasma isolate obtained from honeybee in China and its basic properties

    Institute of Scientific and Technical Information of China (English)

    李霞; 张杰; 马云龙; 于汉寿

    2012-01-01

    [目的]通过分析分离自我国患病蜜蜂体内的螺原体MF1006的基本生物学特征,初步确定其分类地位及致病性.[方法]应用暗视野显微镜和透射电子显微镜观察螺原体形态,运用常规螺原体分离和培养方法、分子生物学方法和血清学方法研究螺原体分离菌株可能的分类地位,并采用饲喂接种法研究其致病性.[结果]菌株MF 1006具有典型的螺旋形和运动性,能通过0.22μm孔径的滤膜,与我国之前发现的引起蜜蜂螺原体病的参比菌株Spiroplasma melliferum CH-1的基本生物学特征差异较大.S.melliferum CH-1抗血清对其没有抑制作用.根据16S rDNA、ITS序列构建系统发育树显示,菌株MF1006与在法国发现的引起蜜蜂“五月病”的Spiroplasma apis的亲缘关系最近.此外,菌株MF1006对供试意蜂有较强的致病性.[结论]分离菌株MF1006是在我国蜜蜂体内发现的除S.melliferum以外的另一种致病螺原体.%[Objective] The purpose of this study was to characterize the isolate MF1006, andto determine the taxonomy and pathogenicity of the spiroplasma obtained from diseased honeybees in China. [Methods] The spiroplasma morphology was examined by dark-field microscope and transmission electron microscope. The biological characteristic of the spiroplasma was investigated by using conventional culture-dependent method and molecular biology and serological methods. We determined the spiroplasma pathogenicity through feeding tests. [Results] MF1006 exhibited typical properties of spiral morphology and mobility. It was able to pass through membrane filters with pores size of 220 nm. This isolate was very different from the previous reported domestic isolate Spiroplasma melliferum CH-1 which caused honeybee spiroplasmosis seriously. S. Melliferum CH-1 antiserum could not inhibit MF1006. Phylogenetic analysis based on the 16S rDNA and ITS revealed that MF1006 was close to S. Apis which was found in France and associated

  1. Nesting lizards (Bassiana duperreyi) compensate partly, but not completely, for climate change.

    Science.gov (United States)

    Telemeco, Rory S; Elphick, Melanie J; Shine, Richard

    2009-01-01

    Species in which ambient temperatures directly determine offspring sex may be at particular risk as global climates change. Whether or not climate change affects sex ratio depends upon the effectiveness of buffering mechanisms that link ambient regimes to actual nest temperatures. For example, females may simply lay nests earlier in the season, or in more shaded areas, such that incubation thermal regimes are unchanged despite massive ambient fluctuation. Based on eight years of monitoring nests over a 10-year period in the field at an alpine site in southeastern Australia, we show that, even though lizards (Bassiana duperreyi, Scincidae) have adjusted both nest depth and seasonal timing of oviposition in response to rising ambient temperatures, they have been unable to compensate entirely for climate change. That inability stems from the fact that the seasonal progression of soil temperatures, and thus, the degree to which thermal regimes at the time of laying predict subsequent conditions during incubation, also has shifted with climate change. As a result, mean incubation temperatures in natural nests now have crossed the thermal threshold at which incubation temperature directly affects offspring sex in this population.

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

    Science.gov (United States)

    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.

  3. The production and uses of Beauveria bassiana as a microbial insecticide.

    Science.gov (United States)

    Mascarin, Gabriel Moura; Jaronski, Stefan T

    2016-11-01

    Among invertebrate fungal pathogens, Beauveria bassiana has assumed a key role in management of numerous arthropod agricultural, veterinary and forestry pests. Beauveria is typically deployed in one or more inundative applications of large numbers of aerial conidia in dry or liquid formulations, in a chemical paradigm. Mass production is mainly practiced by solid-state fermentation to yield hydrophobic aerial conidia, which remain the principal active ingredient of mycoinsecticides. More robust and cost-effective fermentation and formulation downstream platforms are imperative for its overall commercialization by industry. Hence, where economics allow, submerged liquid fermentation provides alternative method to produce effective and stable propagules that can be easily formulated as dry stable preparations. Formulation also continues to be a bottleneck in the development of stable and effective commercial Beauveria-mycoinsecticides in many countries, although good commercial formulations do exist. Future research on improving fermentation and formulation technologies coupled with the selection of multi-stress tolerant and virulent strains is needed to catalyze the widespread acceptance and usefulness of this fungus as a cost-effective mycoinsecticide. The role of Beauveria as one tool among many in integrated pest management, rather than a stand-alone management approach, needs to be better developed across the range of crop systems. Here, we provide an overview of mass-production and formulation strategies, updated list of registered commercial products, major biocontrol programs and ecological aspects affecting the use of Beauveria as a mycoinsecticide.

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

    Science.gov (United States)

    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.

  5. The isolation and identification of pathogenic fungi from Tessaratoma papillosa Drury (Hemiptera: Tessaratomidae

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    Xiang Meng

    2017-10-01

    Full Text Available Background Litchi stink-bug, Tessaratoma papillosa Drury (Hemiptera: Tessaratomidae, is one of the most widespread and destructive pest species on Litchi chinensis Sonn and Dimocarpus longan Lour in Southern China. Inappropriate use of chemical pesticides has resulted in serious environmental problems and food pollution. Generating an improved Integrated Pest Management (IPM strategy for litchi stink-bug in orchard farming requires development of an effective biological control agent. Entomopathogenic fungi are regarded as a vital ecological factor in the suppression of pest populations under field conditions. With few effective fungi and pathogenic strains available to control litchi stink-bug, exploration of natural resources for promising entomopathogenic fungi is warranted. Methods & Results In this study, two pathogenic fungi were isolated from cadavers of adult T. papillosa. They were identified as Paecilomyces lilacinus and Beauveria bassiana by morphological identification and rDNA-ITS homogeneous analysis. Infection of T. papillosa with B. bassiana and P. lilacinus occurred initially from the antennae, metameres, and inter-segmental membranes. Biological tests showed that the two entomopathogenic fungi induced high mortality in 2nd and 5th instar nymphs of T. papillosa. B. bassiana was highly virulent on 2nd instar nymphs of T. papillosa, with values for cadaver rate, LC50 and LT50 of 88.89%, 1.92 × 107 conidia/mL and 4.34 days respectively. Discussion This study provides two valuable entomopathogenic fungi from T. papillosa. This finding suggests that the highly virulent P. lilacinus and B. bassiana play an important role in the biocontrol of T. papillosa in China. These pathogenic fungi had no pollution or residue risk, and could provide an alternative option for IPM of litchi stink-bug.

  6. 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....

  7. Application of hydrophilic-lypophilic balance (HLB) number to optimize a compatible non-ionic surfactant for dried aerial conidia of Beauveria bassiana

    Science.gov (United States)

    The hydrophilic-lipophilic balance (HLB) number system was used to optimize a compatible non-ionic surfactant, TDA(polyoxyethylene tridecyl ether) in formulations for two Beauveria bassiana strains, NI8 and GHA. The optimal HLB number for TDA was determined on the basis of wetting times for conidia...

  8. Impact of Beauveria bassiana and imidacloprid, alone and in combination, used against emerald ash borer in a newly-infested ash nursery

    Science.gov (United States)

    We are investigating the potential of Beauveria bassiana (strain GHA), alone or in combination with imidacloprid, for use against the emerald ash borer (EAB), Agrilus planipennis. We treated approximately 400 Fraxinus pennsylvanica and F. americana (height ca. 5-6 m) at a commercial tree nursery wit...

  9. 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...

  10. Comparative efficacy of emulsifiable-oil, wettable-powder, and unformulated-powder preparations of Beauveria bassiana against the melon aphid Aphis gossypii

    Science.gov (United States)

    Aphicidal efficacy of two commercial formulations of Beauveria bassiana strain GHA conidia, an emulsifiable-paraffinic oil dispersion (OD) and a clay-based wettable powder (WP), were compared to unformulated conidia in laboratory tests with adult melon aphids (Aphis gossypii). For the initial 24 hou...

  11. Two strains of Pseudomonas fluorscens bacteria differentially affect survivorship of waxworm (Galleria mellonella) larvae exposed to an arthropod fungal pathogen, Beauveria bassiana

    Science.gov (United States)

    Two strains of Pseudomonas fluorescens were found contaminating a biopesticide used in a previous study against Varroa destructor infestations in honey bee hives. In the aforementioned study the biopesticide, a formulation of the arthropod pathogen Beauveria bassiana, failed to have any impact on t...

  12. Potential of a strain of the entomopathogenic fungus Beauveria bassiana (Hypocreales: Cordycipitaceae) as a biological control agent against western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae)

    Science.gov (United States)

    Five Beauveria bassiana strains were evaluated for control of western flower thrips. Strain RSB was the most virulent, causing 69-96% mortality at concentrations of 1×104 – 1×107 conidia mL-1, 10 days after inoculation of first instars. In greenhouse trials, RSB applied to broccoli foliage signifi...

  13. Use of a mustard trap crop to study efficacy of Beauveria bassiana treatments to corn and soybean for control of tarnished plant bug in cotton

    Science.gov (United States)

    A preliminary field study was conducted in 2012 to measure impact of spraying tassel-stage corn and blooming early-maturity-group soybean with Beauveria bassiana on tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) populations before they colonize cotton. Four-row strips of mustard (total...

  14. Use of acoustic technology to monitor the time course of Rhynchophorus ferrugineus larval mortality in date palms after treatments with Beauveria bassiana

    Science.gov (United States)

    Spectral and temporal patterns of insect sound impulses were monitored daily for 23-d periods in 8, 10, or 5 small date palm trees containing larvae dipped in 0 (control), 104 (low), or 108 (high) conidia/ml doses of entomopathogenic fungus, Beauveria bassiana (Bb 203), respectively. Each tree conta...

  15. A case of Beauveria bassiana keratitis confirmed by internal transcribed spacer and LSU rDNA D1–D2 sequencing

    Directory of Open Access Journals (Sweden)

    M. Ligozzi

    2014-05-01

    Full Text Available We describe a case of fungal keratitis due to Beauveria bassiana in a farmer with Fuchs' dystrophy, treated with amphotericin B. Surgery with penetrating keratoplasty was necessary to resolve the lesions. Susceptibility testing and molecular sequencing permitted the identification and treatment of this rare aetiological agent of invasive fungal disease.

  16. Field evaluation on the lethal effect of Beauveria bassiana strains NI8 and GHA against the tarnished plant bug in cotton

    Science.gov (United States)

    The entomopathogenic fungus Beauveria bassiana Delta native strain NI8 have shown great potential for the management of tarnished plant bug adults when compared with the commercial strain GHA. Population of L lineolaris in cages was reduced by 50% 10 days after application of the NI8 native strain a...

  17. Repellent activity of desiccant dusts and conidia of the entomopathogenic fungus Beauveria bassiana when tested against poultry red mites (Dermanyssus gallinae) in laboratory experiments

    DEFF Research Database (Denmark)

    Kilpinen, Ole; Steenberg, Tove

    2016-01-01

    on both dose and relative humidity (RH). However, one desiccant dust was significantly less repellent compared to other dusts with similar levels of efficacy. Further, dry conidia of the fungus Beauveria bassiana were also shown to be repellent to poultry red mites, both when applied on its own and when...

  18. Delayed efficacy of Beauveria bassiana foliar spray applications against Colorado potato beetle: impacts of number and timing of applications on larval and next-generation adult populations

    Science.gov (United States)

    Spray programs comprising multiple or single foliar applications of the fungal pathogen Beauveria bassiana strain GHA (Bb) made during morning (AM) vs. evening (PM) hours were tested against Colorado potato beetle Leptinotarsa decemlineata (CPB) in small research plots of potatoes over multiple fiel...

  19. A case of Beauveria bassiana keratitis confirmed by internal transcribed spacer and LSU rDNA D1-D2 sequencing.

    Science.gov (United States)

    Ligozzi, M; Maccacaro, L; Passilongo, M; Pedrotti, E; Marchini, G; Koncan, R; Cornaglia, G; Centonze, A R; Lo Cascio, G

    2014-05-01

    We describe a case of fungal keratitis due to Beauveria bassiana in a farmer with Fuchs' dystrophy, treated with amphotericin B. Surgery with penetrating keratoplasty was necessary to resolve the lesions. Susceptibility testing and molecular sequencing permitted the identification and treatment of this rare aetiological agent of invasive fungal disease.

  20. Compatibility between the entomopathogenic fungus Beauveria bassiana and insecticides used in coffee plantations Compatibilidade entre o fungo entomopatogênico Beauveria bassiana e inseticidas usados na cultura do cafeeiro

    Directory of Open Access Journals (Sweden)

    Carolina Natali de Oliveira

    2003-12-01

    Full Text Available Microbial control in integrated pest management (IPM programs of coffee plantations is an important factor for the reduction of pest population densities. The use of selective pesticides can be associated with entomopathogens, increasing the efficiency of the control and reducing the use of required insecticides. The in vitro fungitoxic effect of insecticide formulations of Thiamethoxam, Cyfluthrin, Deltamethrin, Alpha-Cypermethrin, Triazophos, Chlorpyrifos, Fenpropathrin and Endosulfan and Beauveria bassiana (CG 425 strain was evaluated at three concentrations (FR = average field recommendation; 0.5 ´ FR and 2 ´ FR. Effects of these products on conidia germination, vegetative growth and sporulation were compared. Only five insecticides, at the FR concentration, promoted conidia viability higher than 60%. Viability should be considered the most important factor to be evaluated since it is the first step of the infection process. The insecticide formulations of Alpha-Cypermethrin, Thiamethoxam and Cyfluthrin caused the lower inhibition level on conidia germination at the two lower concentrations, with no difference in relation to the control. With respect to vegetative growth analysis, Thiamethoxam at the two lower concentrations was not found to cause radial growth inhibition. Thiamethoxam caused the smallest inhibition level with regard to conidia production. The use of Alpha-Cypermethrin and Thiamethoxam formulations in coffee IPM programs for a B. bassiana inoculum conservation strategy are recommended, since these products were compatible with the entomopathogenic fungus Beauveria bassiana (CG 425, an important natural control agent of the coffee berry borer, Hypothenemus hampei.Em programas de Manejo integrado de pragas (MIP deve-se considerar o controle microbiano como um importante fator de redução da densidade populacional de pragas. A utilização de produtos seletivos quando associados a patógenos, pode aumentar a eficiência de

  1. Evaluation of soyscreen in an oil-based formulation for UV protection of Beauveria bassiana conidia.

    Science.gov (United States)

    Behle, Robert W; Compton, David L; Laszlo, Joseph A; Shapiro-Ilan, David I

    2009-10-01

    Soyscreen oil was studied as a formulation ingredient to protect Beauveria bassiana (Balsamo) Vuillemin conidia from UV degradation. Feruloylated soy glycerides, referred to as Soyscreen oil, are biobased UV-absorbing molecules made by combining molecules of soybean oil with ferulic acid. Conidia stored in Soyscreen oil for 28 wk at 25, 30, and 35 degrees C retained viability as well as conidia stored in sunflower oil, demonstrating that Soyscreen did not adversely affect viability with prolonged storage. For samples applied to glass and exposed to simulated sunlight (xenon light), conidia in sunflower oil with or without sunscreens (Soyscreen or oxyl methoxycinnimate) had similar conidia viability after exposure. These oil formulations retained conidia viability better than conidia applied as an aqueous treatment. However, the 10% Soyscreen oil formulation applied to field grown cabbage (Brassica oleracea L.) and bean (Phaseolus vulgaris L.) plants, did not improve residual insecticidal activity compared with aqueous applications of unformulated conidia or two commercial formulations when assayed against Trichoplusia ni (Hübner) larvae. Our results suggest that the oil applications lose UV protection because the oil was absorbed by the leaf. This conclusion was supported in subsequent laboratory exposures of conidia in oil-based formulations with UV screens applied to cabbage leaves or balsa wood, which lost protection as measured by decreased viability of conidia when exposed to simulated sunlight. As a result, additional formulation techniques such as encapsulation to prevent separation of the protective oil from the conidia may be required to extend protection when oil formulations are applied in the field.

  2. Molecular Cloning, Bioinformatic Analysis, and Expression of Bombyx mori Lebocin 5 Gene Related to Beauveria bassiana Infection.

    Science.gov (United States)

    Lü, Dingding; Hou, Chengxiang; Qin, Guangxing; Gao, Kun; Chen, Tian; Guo, Xijie

    2017-01-01

    A full-length cDNA of lebocin 5 (BmLeb5) was first cloned from silkworm, Bombyx mori, by rapid amplification of cDNA ends. The BmLeb5 gene is 808 bp in length and the open reading frame encodes a 179-amino acid hydroxyproline-rich peptide. Bioinformatic analysis results showed that BmLeb5 owns an O-glycosylation site and four RXXR motifs as other lebocins. Sequence similarity and phylogenic analysis results indicated that lebocins form a multiple gene family in silkworm as cecropins. Quantitative real-time PCR analysis revealed that BmLeb5 was highest expressed in the fat body. In the silkworm larvae infected by Beauveria bassiana, the expression level of BmLeb5 was upregulated in the fat body and hemolymph which are the most important immune tissues in silkworm. The recombinant protein of BmLeb5 was for the first time successfully expressed with prokaryotic expression system and purified. There are no reports so far that the expression of lebocins could be induced by entomopathogenic fungus. Our study suggested that BmLeb5 might play an important role in the immune response of silkworm to defend B. bassiana infection. The results also provided helpful information for further studying the lebocin family functioned in antifungal immune response in the silkworm.

  3. Evaluación de Beauveria bassiana para el combate de insectos plaga en maíz almacenado

    Directory of Open Access Journals (Sweden)

    Juan Diego Molina

    2000-01-01

    Full Text Available El objetivo de este estudio fue evaluar la patogenicidad de aislamientos de Beauveria bassiana, para controlar los insectos plaga Prostephanus truncatus y Stophilus zeamais, en mazorcas de maíz almacenado. Se evaluó también la patogenicidad del hongo sobre Teritriosoma nigrescens, depredador natural de P. truncatus. Se mezclaron conidios secos del hongo con kaolinita y se aplicaron a trojas de maíz de 450 mazorcas acomodadas en capas de 50. Se montaron dos ensayos, en el primero se evaluaron aplicaciones del hongo al momento de almacenar el maíz, y en el segundo se evaluaron aplicaciones del hongo realizadas después de almacenar el maíz. La mortalidad de P. truncatus fue cuatro veces mayor en el ensayo donde se aplicó el hongo en forma preventiva. La mortalidad de S. zeamais fue el doble en el tratamiento donde se aplicó el hongo al momento de inocular con P. truncatus. Los niveles de mortalidad por B. bassiana fueron mayores en P. truncatus que en S. zeamais, pero S. zeamais, pero este último mostró un 20% más de infección que el primero

  4. Effection of Ocneridia volxemi Bolivar (Pamphaginae, Orthoptera) hoppers and adults by Beauveria bassiana (Deuteromycotina, hyphomycetes) conidia in an oil formulation.

    Science.gov (United States)

    Bounechada, M; Doumandji, S E

    2004-01-01

    Ocneridia volxemi Bolivar (Pamphaginae, Orthoptera) is one of the important insects pest in the heigh plains of north east of Algeria. Larval and adult stages caused severe damage to the leaves of cereals and an other various crops. Laboratory studies were conducted to the determine the potential of the fungus Beauveria bassiana for the control of the pest of larval and adults. Positive results were obtaved when larvae and adults were sprayed with different concentration of fungus conidia (D1: 2 x 10(6) conidia/ml, D2: 5 x 10(5) conidia/ml, D3: 8 x 10(4) conidia/ml, D4: 10(2) conidia/ml). Concentration at D1 and D2 in an oil formulation showed respectively 100% and 90 % larval mortality at the first day of treatment and 100%-9% adults mortality at the five day of treatement. The results revealed that the rates of infection and mortality were significantly related to the concentration of conidia in the corresponding samples. Beauveria bassiana can be considerd as a promising biocontrol agents of Ocneridia volxemi may be an other locuts, but field experimentation is necessary to subtantiate these finding.

  5. The Metapleural Secretion of Acromyrmex laticeps (Forel does not have Fungicide Effect on the Entomopathogenic Fungus Beauveria bassiana (Bals. Vuill

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    Luciane Junqueira

    2014-12-01

    Resumo. A glândula metapleural (GMP é uma estrutura exógena, encontrada apenas nos formicídeos, sendo mais desenvolvida nas fêmeas e operárias, enquanto nos machos ou está atrofiada ou não existe. Tem sido sugerido várias possíveis funções para a secreção produzida por esta glândula, sendo a hipótese de efeito protetor contra entomopatógenos a mais estudada. Para testar esta hipótese, foi aplicada uma suspensão de conídios (1,30 x 106 de Beauveria bassiana (Bals. Vuill. nas GMP e na porção ântero-superior do metatórax (ASM de fêmeas aladas de Acromyrmex laticeps (Forel, enquanto nos machos a suspensão foi aplicada apenas na porção ASM, pois estes não apresentam o átrio característico da GMP. Os resultados obtidos indicaram que a secreção metapleural das fêmeas desta espécie não possui efeito fungicida sobre B. bassiana, e que tanto os machos como as fêmeas alados são sensíveis a este fungo entomopatogênico.

  6. Molecular Cloning, Bioinformatic Analysis, and Expression of Bombyx mori Lebocin 5 Gene Related to Beauveria bassiana Infection

    Science.gov (United States)

    Lü, Dingding; Hou, Chengxiang; Qin, Guangxing; Gao, Kun; Chen, Tian

    2017-01-01

    A full-length cDNA of lebocin 5 (BmLeb5) was first cloned from silkworm, Bombyx mori, by rapid amplification of cDNA ends. The BmLeb5 gene is 808 bp in length and the open reading frame encodes a 179-amino acid hydroxyproline-rich peptide. Bioinformatic analysis results showed that BmLeb5 owns an O-glycosylation site and four RXXR motifs as other lebocins. Sequence similarity and phylogenic analysis results indicated that lebocins form a multiple gene family in silkworm as cecropins. Quantitative real-time PCR analysis revealed that BmLeb5 was highest expressed in the fat body. In the silkworm larvae infected by Beauveria bassiana, the expression level of BmLeb5 was upregulated in the fat body and hemolymph which are the most important immune tissues in silkworm. The recombinant protein of BmLeb5 was for the first time successfully expressed with prokaryotic expression system and purified. There are no reports so far that the expression of lebocins could be induced by entomopathogenic fungus. Our study suggested that BmLeb5 might play an important role in the immune response of silkworm to defend B. bassiana infection. The results also provided helpful information for further studying the lebocin family functioned in antifungal immune response in the silkworm. PMID:28194425

  7. Influence of different types of Phyllostachys pubescens (Poales: Poaceae) leaves on population parameters of Pantana phyllostachysae (Lepidoptera: Lymantriidae) and parasitic effects of Beauveria bassiana (Moniliales: Moniliaceae).

    Science.gov (United States)

    Su, Jun; Zhang, Fei-Ping; Huang, Wen-Ling; Chen, De-Lan; Chen, Shun-Li

    2015-01-01

    We found that Pantana phyllostachysae, a dangerous pest of moso bamboo (Phyllostachys pubescens), showed differences in growth and development after feeding on diverse types of moso bamboo leaves. The mortality rate of Pa. phyllostachysae due to Beauveria bassiana, an entomopathogenic fungus, was also affected by the varied larval diet. Larval and pupal developmental duration of Pa. phyllostachysae was longer when feeding on "off-year" bamboo leaves. Pupal weight and adult fertility were higher when feeding on "on-year" bamboo leaves. Mortality due to B. bassiana was significantly lower in larvae fed on on-year bamboo leaves than in larvae fed on off-year bamboo leaves. Larvae fed on new bamboo leaves had a shorter development period and higher survival rate than those fed on off-year bamboo leaves. However, mixed feed (mixture of new, on-year, and off-year bamboo leaves) decreased the egg production of Pa. phyllostachysae. After infection by the second generation of B. bassiana, the survival time of Pa. phyllostachysae fed on mixed feed increased significantly compared with the first generation. We also fed Pa. phyllostachysae different proportion of new bamboo leaves in mixed feed to simulate natural conditions. We found that increasing the proportion of new bamboo leaves in the food promoted pupal development and increased egg production; it also increased the resistance of larvae to the first generation of B. bassiana. The pathogenicity of the second generation of B. bassiana declined in all mixed feed treatments. © The Author 2015. Published by Oxford University Press on behalf of the Entomological Society of America.

  8. Entomopathogens Isolated from Invasive Ants and Tests of Their Pathogenicity

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    Maria Fernanda Miori de Zarzuela

    2012-01-01

    Full Text Available Some ant species cause severe ecological and health impact in urban areas. Many attempts have been tested to control such species, although they do not always succeed. Biological control is an alternative to chemical control and has gained great prominence in research, and fungi and nematodes are among the successful organisms controlling insects. This study aimed to clarify some questions regarding the biological control of ants. Invasive ant species in Brazil had their nests evaluated for the presence of entomopathogens. Isolated entomopathogens were later applied in colonies of Monomorium floricola under laboratory conditions to evaluate their effectiveness and the behavior of the ant colonies after treatment. The entomopathogenic nematodes Heterorhabditis sp. and Steinernema sp. and the fungi Beauveria bassiana, Metarhizium anisopliae, and Paecilomyces sp. were isolated from the invasive ant nests. M. floricola colonies treated with Steinernema sp. and Heterorhabditis sp. showed a higher mortality of workers than control. The fungus Beauveria bassiana caused higher mortality of M. floricola workers. However, no colony reduction or elimination was observed in any treatment. The defensive behaviors of ants, such as grooming behavior and colony budding, must be considered when using fungi and nematodes for biological control of ants.

  9. Investigation of plasmid profile, antibiotic susceptibility pattern multiple antibiotic resistance index calculation of Escherichia coli isolates obtained from different human clinical specimens at tertiary care hospital in Bareilly-India

    Directory of Open Access Journals (Sweden)

    Ajay Francis Christopher

    2013-01-01

    Full Text Available Background: Escherichia coli is well known as noninvasive commensal and has been established as etiological agent of various human infections. E. coli also contributes to high rate of resistance to several antibiotics due to multiresistant antibiotic plasmid genes e.g., extended spectrum β-lactamases (ESBL. Material and Methods: To analyse the situation of antibiotic resistance, a total of 77 E.coli isolates from urine, pus, sputum and endotracheal aspirate were screened for their antibiograms for antibiotic resistance, multiple antibiotic resistance (MAR index for evaluating the spread of resistance and plasmid profiles for the presence and characterization of plasmids. Results: Very high resistance level (> 90% was detected against ampicillin, amoxycillin, ceftazidime, norfloxacin, tetracycline while imipenem and amikacin recorded the least resistance levels of 2.3% and 13.9%, respectively, among the isolates. An increased resistance to amoxycillin, tetracycline, cotrimoxazole and norfloxacin were observed in this geographical area which however displayed a lower resistance in other countries. The MAR index varied considerably, the lowest was 0.18 and the highest was 0.89. Plasmids of 10 size ranges were detected in the isolates. Some isolates possessed single-sized plasmid while other possessed multiple plasmids. Isolates with high MAR profiles were found to possess multiple plasmids. Conclusion: Regular antimicrobial sensitivity surveillance is necessary and acquisition of plasmid could greatly contribute in the antibiotic resistance and poses a significant risk of the spread of microbial resistance in this community. Also, it was observed that route of administration of antibiotics perhaps reduced its misuse and hence led to the reduction in the emergence of resistant bacterial strains.

  10. Obtaining of inulin acetate

    OpenAIRE

    Khusenov, Arslonnazar; Rakhmanberdiev, Gappar; Rakhimov, Dilshod; Khalikov, Muzaffar

    2014-01-01

    In the article first obtained inulin ester inulin acetate, by etherification of inulin with acetic anhydride has been exposed. Obtained product has been studied using elementary analysis and IR spectroscopy.

  11. Production of extracellular protease by a Brazilian strain of Beauveria bassiana reactivated on coffee berry borer, Hypothenemus hampei

    Directory of Open Access Journals (Sweden)

    Eliana Tiemi Ito

    2007-03-01

    Full Text Available Studies were carried out on extracellular protease production by Beauveria bassiana CG432 in liquid medium containing glucose and yeast extract. B. Bassiana presented active growth after lag period of 24 h., produced 80% of the total of the extracellular protease activity in 48 h which was maximum on the 5th culture day. The extracellular protease presented optimum activity at 60ºC, was stable up to 1M Cl-, maintained the stability during 15 day at 4ºC and -18ºC, but was not stable if frozen repeatedly.A cepa brasileira do fungo entomopatogênico Beauveria bassiana CG432 foi cultivada em meio de cultivo líquido contendo glucose e extrato de levedura a 28°C, 150 rpm, durante 9 dias utilizando 1% de inóculo constituído por 10(8conídios/mL pré-ativados em insetos vivos de broca-do-café. Avaliou-se diariamente o crescimento do fungo através da quantificação da biomassa e a atividade de proteases extracelulares no filtrado do cultivo pela determinação de peptídeos solúveis liberados pela hidrólise de soro albumina bovina a 37°C durante 30minutos. O fungo cresceu ativamente após reduzido período lag de 24 horas, produziu 80% da atividade total de proteases em 48 horas e atingiu produção máxima em 5 dias de cultivo, sem adição de substratos indutores no meio de cultivo. As proteases apresentaram atividade ótima a 60ºC, estabilidade até 1M de íons cloreto, mantiveram mais de 80% de atividade durante 15 dias a 4ºC e -18ºC, porém foram sensíveis a congelamentos repetitivos.

  12. Patogenicidade de Beauveria bassiana ao psilídeo Diaphorina citri e compatibilidade do fungo com produtos fitossanitários Beauveria bassiana pathogenicity to Diaphorina citri and compatibility of the fungus with phytosanitary products

    Directory of Open Access Journals (Sweden)

    Ana Paula Ferreira Pinto

    2012-12-01

    Full Text Available O objetivo deste trabalho foi avaliar a patogenicidade de Beauveria bassiana a ninfas de Diaphorina citri (Hemiptera: Psyllidae e verificar a compatibilidade do fungo com produtos fitossanitários e sua persistência em plantas de citros. Ninfas de D. citri foram pulverizadas com B. bassiana, nas concentrações 5x10(6, 1x10(7, 5x10(7, 1x10(8, 5x10(8 e 1x10(9 conídios mL-1, para determinação da concentração letal. Para avaliação da compatibilidade do fungo com produtos fitossanitários, extrato de nim e cinco inseticidas de quatro grupos químicos diferentes foram incorporados individualmente ao meio de cultura BDA em que o fungo foi cultivado. Avaliaram-se o crescimento vegetativo, a esporulação e a viabilidade do entomopatógeno. Plantas de citros, mantidas em casa de vegetação, foram tratadas primeiramente com os produtos fitossanitários e depois com o entomopatógeno. Avaliaram-se os tempos de exposição de 24 horas e de 7 e 14 dias. O fungo foi patogênico às ninfas de D. citri; a CL50 foi de 0,4x10(7 e a CL90 de 6,7x10(7 conidios mL-1, no décimo dia de avaliação. Em laboratório, os produtos fitosssanitários reduzem o crescimento do fungo. Em casa de vegetação, os produtos não afetam a sobrevivência do fungo nas plantas de citros.The objective of this work was to evaluate the pathogenicity of Beauveria bassiana to Diaphorina citri (Hemiptera: Psyllidae nymphs, and to check the compatibility of the fungus with phytosanitary products, and its persistence in citrus plants. Nymphs of D. citri were sprayed with B. bassiana in the concentrations 5x10(6, 1x10(7, 5x10(7, 1x10(8, 5x10(8 and 1x10(9 conidia mL-1 for lethal concentration determination. In order to evaluate the compatibility of the phytosanitary products with the fungus, neem extract and five insecticides of four different chemical groups were individually added to PDA culture medium, in which the fungus was cultivated. Vegetative growth, sporulation and viability

  13. The entomopathogenic fungus Beauveria bassiana reduces instantaneous blood feeding in wild multi-insecticide-resistant Culex quinquefasciatus mosquitoes in Benin, West Africa

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    Howard Annabel FV

    2010-09-01

    Full Text Available Abstract Background Mosquito-borne diseases are still a major health risk in many developing countries, and the emergence of multi-insecticide-resistant mosquitoes is threatening the future of vector control. Therefore, new tools that can manage resistant mosquitoes are required. Laboratory studies show that entomopathogenic fungi can kill insecticide-resistant malaria vectors but this needs to be verified in the field. Methods The present study investigated whether these fungi will be effective at infecting, killing and/or modifying the behaviour of wild multi-insecticide-resistant West African mosquitoes. The entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana were separately applied to white polyester window netting and used in combination with either a permethrin-treated or untreated bednet in an experimental hut trial. Untreated nets were used because we wanted to test the effect of fungus alone and in combination with an insecticide to examine any potential additive or synergistic effects. Results In total, 1125 female mosquitoes were collected during the hut trial, mainly Culex quinquefasciatus Say. Unfortunately, not enough wild Anopheles gambiae Giles were collected to allow the effect the fungi may have on this malaria vector to be analysed. None of the treatment combinations caused significantly increased mortality of Cx. quinquefasciatus when compared to the control hut. The only significant behaviour modification found was a reduction in blood feeding by Cx. quinquefasciatus, caused by the permethrin and B. bassiana treatments, although no additive effect was seen in the B. bassiana and permethrin combination treatment. Beauveria bassiana did not repel blood foraging mosquitoes either in the laboratory or field. Conclusions This is the first time that an entomopathogenic fungus has been shown to reduce blood feeding of wild mosquitoes. This behaviour modification indicates that B. bassiana could potentially be a new

  14. Application of Molecular Typing Results in Source Attribution Models: The Case of Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) of Salmonella Isolates Obtained from Integrated Surveillance in Denmark

    DEFF Research Database (Denmark)

    de Knegt, Leonardo; Pires, Sara Monteiro; Löfström, Charlotta

    2016-01-01

    number tandem repeat analysis (MLVA) substituted phage typing as the subtyping method for surveillance of S. Enteritidis and S. Typhimurium isolated from animals, food, and humans in Denmark. The purpose of this study was to develop a modeling approach applying a combination of serovars, MLVA types......, and antibiotic resistance profiles for the Salmonella source attribution, and assess the utility of the results for the food safety decisionmakers. Full and simplified MLVA schemes from surveillance data were tested, and model fit and consistency of results were assessed using statistical measures. We conclude...

  15. Isolation and characterization of Wharton’s jelly-derived multipotent mesenchymal stromal cells obtained from bovine umbilical cord and maintained in a defined serum-free three-dimensional system

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    Cardoso Tereza C

    2012-05-01

    Full Text Available Abstract Background The possibility for isolating bovine mesenchymal multipotent cells (MSCs from fetal adnexa is an interesting prospect because of the potential for these cells to be used for biotechnological applications. Bone marrow and adipose tissue are the most common sources of MSCs derived from adult animals. However, little knowledge exists about the characteristics of these progenitors cells in the bovine species. Traditionally most cell cultures are developed in two dimensional (2D environments. In mammalian tissue, cells connect not only to each other, but also support structures called the extracellular matrix (ECM. The three-dimensional (3D cultures may play a potential role in cell biotechnology, especially in tissue therapy. In this study, bovine-derived umbilical cord Wharton’s jelly (UC-WJ cells were isolated, characterized and maintained under 3D-free serum condition as an alternative of stem cell source for future cell banking. Results Bovine-derived UC-WJ cells, collected individually from 5 different umbilical cords sources, were successfully cultured under serum-free conditions and were capable to support 60 consecutive passages using commercial Stemline® mesenchymal stem cells expansion medium. Moreover, the UC-WJ cells were differentiated into osteocytes, chondrocytes, adipocytes and neural-like cells and cultured separately. Additionally, the genes that are considered important embryonic, POU5F1 and ITSN1, and mesenchymal cell markers, CD105+, CD29+, CD73+ and CD90+ in MSCs were also expressed in five bovine-derived UC-WJ cultures. Morphology of proliferating cells typically appeared fibroblast-like spindle shape presenting the same viability and number. These characteristics were not affected during passages. There were 60 chromosomes at the metaphase, with acrocentric morphology and intense telomerase activity. Moreover, the proliferative capacity of T cells in response to a mitogen stimulus was suppressed when

  16. 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

    Directory of Open Access Journals (Sweden)

    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.

  17. Differential contributions of five ABC transporters to mutidrug resistance, antioxidion and virulence of Beauveria bassiana, an entomopathogenic fungus.

    Science.gov (United States)

    Song, Ting-Ting; Zhao, Jing; Ying, Sheng-Hua; Feng, Ming-Guang

    2013-01-01

    Multidrug resistance (MDR) confers agrochemical compatibility to fungal cells-based mycoinsecticdes but mechanisms involved in MDR remain poorly understood for entomopathogenic fungi, which have been widely applied as biocontrol agents against arthropod pests. Here we characterized the functions of five ATP-binding cassette (ABC) transporters, which were classified to the subfamilies ABC-B (Mdr1), ABC-C (Mrp1) and ABC-G (Pdr1, Pdr2 and Pdr5) and selected from 54 full-size ABC proteins of Beauveria bassiana based on their main domain architecture, membrane topology and transcriptional responses to three antifungal inducers. Disruption of each transporter gene resulted in significant reduction in resistance to four to six of eight fungicides or antifungal drugs tested due to their differences in structure and function. Compared with wild-type and complemented (control) strains, disruption mutants of all the five transporter genes became significantly less tolerant to the oxidants menadione and H₂O₂ based on 22-41% and 10-31% reductions of their effective concentrations required for the suppression of 50% colony growth at 25°C. Under a standardized spray, the killing actions of ΔPdr5 and ΔMrp1 mutants against Spodoptera litura second-instar larvae were delayed by 59% and 33% respectively. However, no significant virulence change was observed in three other delta mutants. Taken together, the examined five ABC transporters contribute differentially to not only the fungal MDR but antioxidant capability, a phenotype rarely associated with ABC efflux pumps in previous reports; at least some of them are required for the full virulence of B. bassiana, thereby affecting the fungal biocontrol potential. Our results indicate that ABC pump-dependent MDR mechanisms exist in entomopathogenic fungi as do in yeasts and human and plant pathogenic fungi.

  18. Differential contributions of five ABC transporters to mutidrug resistance, antioxidion and virulence of Beauveria bassiana, an entomopathogenic fungus.

    Directory of Open Access Journals (Sweden)

    Ting-Ting Song

    Full Text Available Multidrug resistance (MDR confers agrochemical compatibility to fungal cells-based mycoinsecticdes but mechanisms involved in MDR remain poorly understood for entomopathogenic fungi, which have been widely applied as biocontrol agents against arthropod pests. Here we characterized the functions of five ATP-binding cassette (ABC transporters, which were classified to the subfamilies ABC-B (Mdr1, ABC-C (Mrp1 and ABC-G (Pdr1, Pdr2 and Pdr5 and selected from 54 full-size ABC proteins of Beauveria bassiana based on their main domain architecture, membrane topology and transcriptional responses to three antifungal inducers. Disruption of each transporter gene resulted in significant reduction in resistance to four to six of eight fungicides or antifungal drugs tested due to their differences in structure and function. Compared with wild-type and complemented (control strains, disruption mutants of all the five transporter genes became significantly less tolerant to the oxidants menadione and H₂O₂ based on 22-41% and 10-31% reductions of their effective concentrations required for the suppression of 50% colony growth at 25°C. Under a standardized spray, the killing actions of ΔPdr5 and ΔMrp1 mutants against Spodoptera litura second-instar larvae were delayed by 59% and 33% respectively. However, no significant virulence change was observed in three other delta mutants. Taken together, the examined five ABC transporters contribute differentially to not only the fungal MDR but antioxidant capability, a phenotype rarely associated with ABC efflux pumps in previous reports; at least some of them are required for the full virulence of B. bassiana, thereby affecting the fungal biocontrol potential. Our results indicate that ABC pump-dependent MDR mechanisms exist in entomopathogenic fungi as do in yeasts and human and plant pathogenic fungi.

  19. A novel method to optimize culture conditions for biomass and sporulation of the entomopathogenic fungus Beauveria Bassiana IBC1201

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    Li Gao

    2011-12-01

    Full Text Available Biomass yields and sporulation of Beauveria bassiana was concerned on culture conditions, environmental factors and cultivation method. We optimized the best culture conditions for biomass yields of B. bassiana IBC1201 with the novel "two-stage" cultivation method as well as orthogonal matrix method. Firstly, we cultured spore suspension on the basal medium (sucrose 19.00 g, soy peptone 4.06 g, K2HPO4 1.00 g, KCl 0.50 g, MgSO4 0.50 g, FeSO4 0.10 g and 17.00 g Bactor for the first stage culture of 4 days under room condition. Then, we transferred them to another defined medium (Cellobiose 9.52 g, urea 1.70 g, ZnSO4•7H2O 0.05 g/L, MnSO4•H2O 0.005 g/L, CaCl2 1.00 g/L, CuSO4•5H2O 0.05 g/L and 17.00 g Bactor for more 4 days cultivation with the environmental factors combination of water potential -1.2 MPa /pH 3 /12 h light cycle/ 23 ℃ for biomass yields, and with the environmental factors combination of water potential -0.8 MPa /pH 3 /24 h light cycle/ 23 ℃ for spore yields. These results provided important information for mass production (including biomass and spore yields of this great potential biocontrol fungus.

  20. A novel method to optimize culture conditions for biomass and sporulation of the entomopathogenic fungus Beauveria bassiana IBC1201.

    Science.gov (United States)

    Gao, Li

    2011-10-01

    Biomass yields and sporulation of Beauveria bassiana was concerned on culture conditions, environmental factors and cultivation method. We optimized the best culture conditions for biomass yields of B. bassiana IBC1201 with the novel "two-stage" cultivation method as well as orthogonal matrix method. Firstly, we cultured spore suspension on the basal medium (sucrose 19.00 g, soy peptone 4.06 g, K2HPO4 1.00 g, KCl 0.50 g, MgSO4 0.50 g, FeSO4 0.10 g and 17.00 g Bactor) for the first stage culture of 4 days under room condition. Then, we transferred them to another defined medium (Cellobiose 9.52 g, urea 1.70 g, ZnSO4•7H2O 0.05 g/L, MnSO4•H2O 0.005 g/L, CaCl2 1.00 g/L, CuSO4•5H2O 0.05 g/L and 17.00 g Bactor) for more 4 days cultivation with the environmental factors combination of water potential -1.2 MPa /pH 3 /12 h light cycle/23 ℃ for biomass yields, and with the environmental factors combination of water potential -0.8 MPa /pH 3 /24 h light cycle/23 ℃ for spore yields. These results provided important information for mass production (including biomass and spore yields) of this great potential biocontrol fungus.

  1. Growth substrates and caleosin-mediated functions affect conidial virulence in the insect pathogenic fungus Beauveria bassiana.

    Science.gov (United States)

    Ortiz-Urquiza, Almudena; Fan, Yanhua; Garrett, Timothy; Keyhani, Nemat O

    2016-11-01

    The entomopathogenic fungus, Beauveria bassiana, is a microbial biological control agent capable of infecting a wide range of insect hosts. Conidia (spores) initiate infection via adhesion, growth and penetration of the insect cuticle, whose outmost layer is rich in lipids. Conidial virulence was investigated in B. bassiana WT and caleosin mutants (ΔBbcal1), the latter a protein involved in lipid storage and turnover. Topical insect bioassays revealed that conidia of the WT strain showed up to 40-fold differences in LD50 values depending upon the growth substrate. The most virulent conidia were harvested from potato dextrose agar containing oleic acid, and the least potent were those derived from Sabouraud dextrose/yeast extract agar (SDAY). However, with the exception of conidia derived from SDAY and Czapek Dox agar, in which values were reduced, mean lethal times to kill (LT50) were essentially unaffected. In topical bioassays, the ΔBbcal1 mutant displayed LD50 values 5-40-fold higher than the WT depending upon the growth substrate, with ΔBbcal1 conidia derived from SDAY unable to effectively penetrate the host cuticle. The ΔBbcal1 mutant also showed concomitant dramatic increases in LT50 values from a mean of ~4.5 for WT to >8.5 days for the mutant. In contrast, intrahaemocoel injection bioassays that bypass cuticle penetration events revealed only minor effects on virulence for either WT or ΔBbcal1 conidia. These data highlight the importance of caleosin-dependent lipid mobilization and/or signalling in cuticle penetration events but suggest their dispensability for immune evasion and within-host growth.

  2. First insight into genetic diversity of the Mycobacterium tuberculosis complex in Albania obtained by multilocus variable-number tandem-repeat analysis and spoligotyping reveals the presence of beijing multidrug-resistant isolates.

    Science.gov (United States)

    Tafaj, Silva; Zhang, Jian; Hauck, Yolande; Pourcel, Christine; Hafizi, Hasan; Zoraqi, Grigor; Sola, Christophe

    2009-05-01

    We characterized a set of 100 Mycobacterium tuberculosis complex clinical isolates from tuberculosis (TB) patients in Albania, typing them with a 24-locus variable-number tandem-repeat-spoligotyping scheme. Depending on the cluster definition, 43 to 49 patients were distributed into 15 to 16 clusters which were likely to be epidemiologically linked, indicative of a recent transmission rate of 28 to 34%. This result suggests that TB is under control in Albania. However, two multidrug-resistant (MDR) Beijing genotypes harboring the same S531A mutation on the rpoB gene were also found, suggesting a potential recent transmission of MDR TB. Three brand new genotypes, Albania-1 to Albania-3, are also described.

  3. Tolerance and bioaccumulation of copper by the entomopathogen Beauveria bassiana (Bals.-Criv.) Vuill. exposed to various copper-based fungicides.

    Science.gov (United States)

    Martins, Fátima; Soares, Maria Elisa; Oliveira, Ivo; Pereira, José Alberto; de Lourdes Bastos, Maria; Baptista, Paula

    2012-07-01

    This work evaluates for the first time the relationships between copper-tolerance, -solubilization and -bioaccumulation in the entomopathogen Beauveria bassiana exposed to Bordeaux mixture, copper oxychloride or copper hydroxide. Bordeaux mixture was highly detrimental to fungus, by inhibiting the growth totally at the recommended dose (RD) and 2×RD. Copper hydroxide and copper oxychloride were found to be less toxic, reducing fungus growth, sporulation and conidial germination in an average of 29  %, 30 % and 58 %, respectively. These two copper forms were the easiest to solubilize, to precipitate and the most accumulated by B. bassiana, suggesting the involvement of all these processes on fungus copper-tolerance.

  4. 湖北省白僵菌研究与应用进展%Progress in Research and Application of Beauveria bassiana in Hubei Province

    Institute of Scientific and Technical Information of China (English)

    江厚利; 查玉平; 陈京元; 赵东容

    2012-01-01

    Beauveria bassiana(Bals.) Vuillemin is a type of entomopathogenic fungi which applied widely in agroforestry pest control.B.bassiana has been applied in forest pest control in Hubei Province Since 1956.Research of B.bassiana basic biology and application in Hubei Province over the past decades was reviewed by authors.%白僵菌是1类广泛应用于农林害虫生物防治的昆虫病原真菌。早在1956年湖北省就开始试用白僵菌防治林业害虫。笔者对湖北省几十年来在白僵菌的基础生物学和应用方面的研究进行了综述。

  5. Conidia production by Beauveria bassiana (for the biocontrol of a diamondback moth) during solid-state fermentation in a packed-bed bioreactor.

    Science.gov (United States)

    Kang, S W; Lee, S H; Yoon, C S; Kim, S W

    2005-01-01

    Conidia of Beauveria bassiana CS-1, which have the potential for the control of the diamondback moth (Plutella xylostella), were produced by solid-state fermentation (SSF) using a packed-bed bioreactor with rice straw and wheat bran. As the packing density and the bed height were increased, the production of conidia decreased. In a packed-bed bioreactor under no aeration and no addition of polypropylene (PP) foam (control), the total average of conidia was 4.9 x 10(8) g-1. The production of conidia was affected more by the addition of PP foam as an inert support than forced aeration and was approx. 23 times higher than that of the control. The total average of conidia produced by B. bassiana was 1.1-1.2 x 10(10) g-1 .

  6. Inhibitory activity of Beauveria bassiana and Trichoderma spp. on the insect pests Xylotrechus arvicola (Coleoptera: Cerambycidae) and Acanthoscelides obtectus (Coleoptera: Chrisomelidae: Bruchinae).

    Science.gov (United States)

    Rodríguez-González, Álvaro; Mayo, Sara; González-López, Óscar; Reinoso, Bonifacio; Gutierrez, Santiago; Casquero, Pedro Antonio

    2017-01-01

    Xylotrechus arvicola is an important pest in vineyards (Vitis vinifera) in the main Iberian wine-producing regions, and Acanthoscelides obtectus causes severe post-harvest losses in the common bean (Phaseolus vulgaris). Under laboratory conditions with a spray tower, the susceptibility of the immature stages of X. arvicola and A. obtectus against the entomopathogenic fungi Beauveria bassiana and four strains of Trichoderma spp. was evaluated. Both insect pests T. harzianum and B. bassiana showed a good inhibitory activity, accumulating an inhibition on the eggs of values above 85 and 82%, respectively. T. atroviride and T. citrinoviride had a lower inhibitory activity, with inhibition values of 74.1 and 73.3% respectively. These fungi can be considered a highly effective tool for the control during the immature stages of these species.

  7. 温度对球孢白僵菌生物学特性的影响%Effects of Temperature on the Growth of Beauveria bassiana

    Institute of Scientific and Technical Information of China (English)

    祁楠

    2013-01-01

      研究了不同温度对球孢白僵菌生长的影响,结果表明在22~26益温度范围内球孢白僵菌落个数随温度的升高而逐渐增多,在26益时达到最大。随后随着温度的升高菌落个数逐渐降低。%This paper researched the influence of different temperatures,The results showed that the Beauveria bassiana colony di-ameter increased with temperature increasing at range of 22~26℃.At 26℃ the colony diameter were the biggest.then the Beauve-ria bassiana colony diameter decreased with temperature decreased.

  8. Study of increasing spore content of Beauveria bassiana%提高白僵菌孢子含量的研究

    Institute of Scientific and Technical Information of China (English)

    刘杰; 陈荣; 孙惠杰

    2001-01-01

    Adding some protein of Antheraea pernyi pupae as the replacement of powder of bean cake and meize flour into Beauveria bassiana raw material and culturing solid in three degrees in iron plate can significantly increase the spores content of Beauveria bassiana,reduce the cost and raise the effect.%在生产白僵菌的原料中,加入一定量的柞蚕蛹蛋白代替豆饼粉或玉米粉,采用铁盘三级固体培养,可显著提高白僵菌的含孢量、缩短生产周期、降低成本、提高对害虫的杀伤速率和效果。

  9. 应用白僵菌防治玉米螟的研究进展%Research Progress of Prevention and Controlling Corn Borer by Beauveria bassiana

    Institute of Scientific and Technical Information of China (English)

    陆晴; 佘花娣; 佟文悦

    2013-01-01

    The damage characteristics of corn borer, biological characteristics and pathogenic mechanism of Beauveria bassiana were introduced briefly.The present situation, problems and corresponding measures of prevention and controlling corn borer using Beauveria bassiana were summarized, and the new direction was discussed so as to promote the corn safety production and improve the control efficiency of corn borer.%简要介绍了玉米螟的为害特点以及白僵菌的生物学特性、致病机理,对利用白僵菌防治玉米螟的现状、存在问题及相应措施进行了阐述,并对防治玉米螟的新方向进行了展望,以促进玉米安全生产和提高玉米螟的防治效率。

  10. Experiment of Beauveria bassiana Against Dung Beetles in Sugarcane%白僵菌防治甘蔗金龟子试验

    Institute of Scientific and Technical Information of China (English)

    薛晶; 张会华; 杨晓丽; 何文志; 罗志明

    2012-01-01

    Experiment of Beauveria bassiana was conducted to control Dung beetles in sugarcane. The results indicated that con- trol effect could be 75% to 85% while applying Beauveria bassiana mixed up with 3.6% of Bisultap granular, which is obviously or significant greatly superior to this agent alone or other pesticides use.%试验结果表明,白僵菌与3.6%广谱型杀虫双颗粒剂混合施用,能有效控制金龟子幼虫对蔗根的危害,相对防效迭75%~85%,显著或极显著优于单独使用白僵菌或农药处理。

  11. Interaction between TATA-Binding Protein (TBP) and Multiprotein Bridging Factor-1 (MBF1) from the Filamentous Insect Pathogenic Fungus Beauveria bassiana

    OpenAIRE

    Chi Song; Almudena Ortiz-Urquiza; Sheng-Hua Ying; Jin-Xia Zhang; Nemat O Keyhani

    2015-01-01

    TATA-binding protein (TBP) is a ubiquitous component of eukaryotic transcription factors that acts to nucleate assembly and position pre-initiation complexes. Multiprotein bridging factor 1 (MBF1) is thought to interconnect TBP with gene specific transcriptional activators, modulating transcriptional networks in response to specific signal and developmental programs. The insect pathogen, Beauveria bassiana, is a cosmopolitan fungus found in most ecosystems where it acts as an important regula...

  12. 森得保与白僵菌防治刚竹毒蛾对比试验%Effects of Sendebao and Beauveria Bassiana on Controlling Pantana phyllostachysae

    Institute of Scientific and Technical Information of China (English)

    詹祖仁; 陈锡桓; 张龙华; 张登强; 叶春珍

    2009-01-01

    采用森得保和白僵菌(Beauveria bassiana)两种生物农药在林间对刚竹毒蛾(Pantana phyllostachysae Chao)3~4龄幼虫进行防治对比试验,结果表明,防治效果分别为94.6%和59.4%,前者极显著高于后者.

  13. Distinct roles of two cytoplasmic thioredoxin reductases (Trr1/2) in the redox system involving cysteine synthesis and host infection of Beauveria bassiana.

    Science.gov (United States)

    Zhang, Long-Bin; Tang, Li; Ying, Sheng-Hua; Feng, Ming-Guang

    2016-12-01

    Two thioredoxin (Trx) reductases (Trr1/2) are known to play overlapping roles in the yeast Trx-Trr redox system but are generally unexplored in filamentous fungi, which possess multiple Trx homologues. This study seeks to characterize the functions of Trr1 and Trr2 in Beauveria bassiana, a filamentous fungal insect pathogen, and to probe their Trx partners. Both Trr1 and Trr2 were evidently localized in the cytoplasm of B. bassiana, unlike the two yeast homologues that have been reported to localize in the cytoplasm and mitochondria, respectively. Most of the six trx genes were greatly upregulated at the transcriptional level in the absence of trr1 instead of trr2 in B. bassiana, in which the trr1/2 double deletion failed in many attempts. Deletion of trr1 resulted in increased Trx activity, severe cysteine auxotrophy, and drastically reduced activities of peroxidases and superoxide dismutases under normal or oxidative conditions despite little change in catalase activity. Such changes disappeared in the absence of trr2 and were completely restored by complementation of trr1/2 or overexpression of trx1/6 in the Δtrr1 mutant, but were not restored at all by overexpression of trx2/3/4/5 or trr2 in the same mutant. All of these mutants exhibited similar trends of changes in the antioxidant response, conidiation, germination, thermotolerance, UV-B resistance, and virulence. Taken together, the findings indicate that Trr1 could reduce Trx2-5 and hence dominate the intracellular redox state, profoundly affecting the potential of B. bassiana against arthropod pests. Trr2 could reduce Trx1/6 but function only in the absence of Trr1.

  14. Uji Efektivitas Protein Hidrolisat Ikan Cucut (Carcharhinus sp.), Naktar, dan Agens Hayati Beauveria bassiana Mengendalikan Lalat Buah (Bactrocera sp.) (Diptera: Tephtritidae) di Laboratorium

    OpenAIRE

    Sembiring, Immanuel

    2014-01-01

    The objectives of the research were to study the ability of protein hydrolysis swordfish, nectar and biological agent (Beauveria bassiana) to control fruit flies (Bractocera dorsalis). The research was held at Laboratory of Agricultural Research and Development of Fruit Crops Experimental Garden Plants Berastagi, Kecamatan Dolat Rayat, Kabupaten Karo (altitude 1.340 meters). The method used Randomized Complete Design with two factors and three replications. The first factor was protein hydrol...

  15. Application of Molecular Typing Results in Source Attribution Models: The Case of Multiple Locus Variable Number Tandem Repeat Analysis (MLVA) of Salmonella Isolates Obtained from Integrated Surveillance in Denmark.

    Science.gov (United States)

    de Knegt, Leonardo V; Pires, Sara M; Löfström, Charlotta; Sørensen, Gitte; Pedersen, Karl; Torpdahl, Mia; Nielsen, Eva M; Hald, Tine

    2016-03-01

    Salmonella is an important cause of bacterial foodborne infections in Denmark. To identify the main animal-food sources of human salmonellosis, risk managers have relied on a routine application of a microbial subtyping-based source attribution model since 1995. In 2013, multiple locus variable number tandem repeat analysis (MLVA) substituted phage typing as the subtyping method for surveillance of S. Enteritidis and S. Typhimurium isolated from animals, food, and humans in Denmark. The purpose of this study was to develop a modeling approach applying a combination of serovars, MLVA types, and antibiotic resistance profiles for the Salmonella source attribution, and assess the utility of the results for the food safety decisionmakers. Full and simplified MLVA schemes from surveillance data were tested, and model fit and consistency of results were assessed using statistical measures. We conclude that loci schemes STTR5/STTR10/STTR3 for S. Typhimurium and SE9/SE5/SE2/SE1/SE3 for S. Enteritidis can be used in microbial subtyping-based source attribution models. Based on the results, we discuss that an adjustment of the discriminatory level of the subtyping method applied often will be required to fit the purpose of the study and the available data. The issues discussed are also considered highly relevant when applying, e.g., extended multi-locus sequence typing or next-generation sequencing techniques.

  16. Antifungal suscepitibility profile of candida spp. oral isolates obtained from denture wearers Perfil de sensibilidade antifungica de isolados de candida sp obtidos de usuários de prótese total

    Directory of Open Access Journals (Sweden)

    J.P. Lyon

    2008-12-01

    Full Text Available Denture stomatitis is an inflammatory condition that occurs in denture wearers and is frequently associated with Candida yeasts. Antifungal susceptibility profiles have been extensively evaluated for candidiasis patients or immunosupressed individuals, but not for healthy Candida carriers. In the present study, fluconazole, itraconazole, voriconazole, terbinafine and 5-flucytosin were tested against 109 oral Candida spp. isolates. All antifungal agents were effective against the samples tested except for terbinafine. This work might provide epidemiological information about Candida spp. drug susceptibility in oral healthy individuals.A estomatite protética é uma condição inflamatória que ocorre em usuários de prótese total e está frequentemente associada a leveduras do gênero Candida, Os perfis de suscetibilidade a antifúngicos têm sido extensivamente estudados em pacientes com candidíase ou em indivíduos imunossuprimidos, mas não em portadores sadios de Candida. No presente estudo, fluconazol, itraconazol, voriconazol, terbinafina e 5-flucitosina foram testados contra 109 isolados orais de Candida spp. Todos os agentes antifúngicos mostraram-se eficazes contra as amostras avaliadas, exceto a Terbinafina. O presente trabalho pode fornecer dados epidemiológicos com relação à susceptibilidade a antifúngicos de Candida spp em indivíduos com saúde oral.

  17. Synergistic interaction between Beauveria bassiana- and Bacillus thuringiensis tenebrionis-based biopesticides applied against field populations of Colorado potato beetle larvae.

    Science.gov (United States)

    Wraight, S P; Ramos, M E

    2005-11-01

    Commercial biopesticides based on the fungal pathogen Beauveria bassiana strain GHA and the bacterial pathogen Bacillus thuringiensis tenebrionis were applied alone