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Sample records for duddingtonia flagrans chlamydospores

  1. Induction of traps by Ostertagia ostertagi larvae, chlamydospore production and growth rate in the nematode-trapping fungus Duddingtonia flagrans

    DEFF Research Database (Denmark)

    Grønvold, J.; Nansen, P.; Henriksen, S. A.

    1996-01-01

    Biological control of parasitic nematodes of domestic animals can be achieved by feeding host animals chlamydospores of the nematode-trapping fungus Duddingtonia flagrans. In the host faeces, D. flagrans develop traps that may catch nematode larvae. In experiments on agar, D. flagrans had a growth...

  2. Duddingtonia flagrans chlamydospores in nutritional pellets: effect of storage time and conditions on the trapping ability against Haemonchus contortus larvae.

    Science.gov (United States)

    Fitz-Aranda, J A; Mendoza-de-Gives, P; Torres-Acosta, J F J; Liébano-Hernández, E; López-Arellano, M E; Sandoval-Castro, C A; Quiroz-Romero, H

    2015-01-01

    The study evaluated the effect of storage time and conditions of nutritional pellets (NP) containing Duddingtonia flagrans chlamydospores on its in vitro trapping ability against Haemonchus contortus L3 larvae. The treated batch (200 NP) contained 4 × 106 chlamydospores of the FTH0-8 strain, whereas the control batch (200 NP) was produced without spores. Both NP batches were exposed to four experimental storage conditions: (T1) shelves (indoors); (T2) refrigeration (4°C); (T3) outdoors under a roof; and (T4) 100% outdoors. Each group comprised 48 NP with spores and 48 NP without spores (control). The ability of D. flagrans spores to trap H. contortus L3 larvae was evaluated for 8 weeks for each storage condition. For that purpose, six randomly selected NP with spores were compared to their respective control NP. Each NP was individually crushed. The crushed material (1 g) was placed on the surface of a 2% water agar plate with 200 H. contortus L3 larvae. Plates were sealed and were incubated at room temperature for 8 days. The whole content of every plate was transferred to a Baermann apparatus to recover the remaining larvae. There was a clear larval reduction in the NP with spores, compared to the respective control NP in the four storage conditions (PT2), 77 ± 6.1 (T1), 81.5 ± 3.8 (T4) and 82.1 ± 2.5 (T3). Larval reductions were similar at all times and were not affected by storage conditions or storage time (R 20.05). The long-term shelf-life of the chlamydospores in the NP suggests that this spore dosage technology is a viable option.

  3. Trematodes enhance the development of the nematode-trapping fungus Arthrobotrys (Duddingtonia) flagrans.

    Science.gov (United States)

    Arias, María Sol; Suárez, José; Cazapal-Monteiro, Cristiana Filipa; Francisco, Iván; López-Arellano, María Eugenia; Piñeiro, Pablo; Suárez, José Luis; Sánchez-Andrade, Rita; Mendoza de Gives, Pedro; Paz-Silva, Adolfo

    2013-01-01

    The capability of helminth (nematode and trematode) parasites in stimulating nematode trap and chlamydospore development of the nematophagous fungus Arthrobotrys (formerly Duddingtonia) flagrans was explored. Dead adult specimens of trematodes (the liver fluke Fasciola hepatica and the rumen fluke Calicophoron daubneyi) and nematodes (the ascarid Parascaris equorum and the strongylid Oesophagostomum spp.), as well as their secretory products, were placed onto corn meal agar plates concurrently inoculated with A. flagrans. Trapping organs were observed after 5 d and chlamydospores after 16 d, including in the control plates in the absence of parasitic stimulus. However, our data shows that both nematodes and trematodes increase trap and chlamydospore production compared with controls. We show for the first time that significantly higher numbers of traps and chlamydospores were observed in the cultures coinoculated with adult trematodes. We conclude that both the traps and chlamydospores formation are not only related to nematode-specific stimuli. The addition of secretory products of the trematode C. daubneyi to culture medium has potential for use in the large scale production of chlamydospores. Copyright © 2013 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  4. Efficacy of an energy block containing Duddingtonia flagrans in the control of gastrointestinal nematodes of sheep.

    Science.gov (United States)

    Sagüés, María F; Fusé, Luis A; Fernández, Alicia S; Iglesias, Lucía E; Moreno, Fabiana C; Saumell, Carlos A

    2011-09-01

    The efficacy of the nematode-trapping fungus Duddingtonia flagrans incorporated into an energy block was evaluated for the control of gastrointestinal nematodes in sheep. Four naturally parasitised sheep with average nematode egg counts of 2,470 eggs per gram grazed by pairs on two similar parasite-free paddocks for 30 days. During that period, one pair of sheep (treated animals, T1) received an energy block containing chlamydospores of D. flagrans at a dose of 200,000 chlamydopores/kg bw/day, while the second pair (control animals, C1) received a fungus-free energy block. The animals in both groups were taken off the paddocks after contaminating the pastures for a month with either nematode eggs plus fungal chlamydospores (T1) or nematode eggs alone (C1). Twelve parasite-free sheep were divided into two groups of six animals each, the treated group (T2) was placed on the paddock previously contaminated with parasites and fungus, while the control group (C2) was placed on the parasite-only paddock. These two groups grazed on their respective paddocks during 30 days and were then housed for 15 days, after which period they were slaughtered in order to determine the parasite burden present in each animal. Results showed that animals in group T2 harboured significantly less nematodes than their counterpart in group C2. The efficacy of D. flagrans was 92% against the total parasite burden, 100% against Haemonchus contortus and Teladorsagia circumcincta, 89.9% against Trichostrongylus colubriformis, 87.5% against Cooperia onchopora, and 90% against Trichostrongylus axei. No efficacy was detected against Nematodirus spathiger, Trichuris ovis and T. skrjabini.

  5. Isolation and Characterization of China Isolates of Duddingtonia flagrans, a Candidate of the Nematophagous Fungi for Biocontrol of Animal Parasitic Nematodes.

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    Wang, Bo-Bo; Liu, Wei; Chen, Ming-Yue; Li, Xuan; Han, Yuan; Xu, Qiang; Sun, Long-Jie; Xie, De-Qiong; Cai, Kui-Zheng; Liu, Yi-Zhong; Liu, Jun-Lin; Yi, Lin-Xin; Wang, Hui; Zhao, Ming-Wang; Li, Xiao-Shan; Wu, Jia-Yan; Yang, Jing; Wang, Yue-Ying

    2015-08-01

    The nematophagous fungus Duddingtonia flagrans has been investigated as a biological agent for the control of gastrointestinal nematodes infecting domestic animals in other countries. However, D. flagrans has not been detected in China. In this study 1,135 samples were examined from 2012 to 2014; 4 D. flagrans isolates (SDH 035, SDH 091, SFH 089, SFG 170) were obtained from the feces of domestic animals and dung compost. The 4 isolates were then characterized morphologically. The SDH 035 strain was characterized by sequencing the ITS1-5.8S rDNA-ITS2 region. A BLAST search showed that the SDH 035 strain (GenBank KP257593) was 100% identical to Arthrobotrys flagrans (AF106520) and was identified as D. flagrans. The morphological plasticity of the isolated strain and the interaction of this strain with the nematode targets were observed by subjecting the infected trichostrongylide L3 to scanning electron microscopy. At 6 and 8 hr after trichostrongylide L(3) was added, hyphal ramifications were observed and L(3) were captured, respectively. Scanning electron micrographs were obtained at 0, 6, 12, 18, 24, 30, 36, 42, and 48 hr, where 0 is the time when trichostrongylide L(3) were first captured by the fungus. The details of the capture process by the fungus are also described. Chlamydospores were observed in the body of L(3) in the late stage of digestion. A sticky substance and bacteria could be observed in contact areas between predation structures and nematode cuticle.

  6. Proteolytic action of the crude extract ofDuddingtonia flagrans on cyathostomins (Nematoda: Cyathostominae in coprocultures

    Directory of Open Access Journals (Sweden)

    Fabio Ribeiro Braga

    Full Text Available The objective of this study was to examine the action of the crude extract of Duddingtonia flagrans (isolates AC001 and CG722 on infective larvae (L3 of cyathostomins in coprocultures and to confirm its proteolytic activity by means of a zymogram. The following groups were formed in coprocultures: Group 1: 10 mL of crude extract of D. flagrans (AC001; group 2: 10 mL of crude extract of AC001 with 10 mM of Ca2+; group 3: 10 mL of crude extract of D. flagrans (CG722; group 4: 10 mL of crude extract of CG722 with 10 mM of Ca2+; and group 5: control group (distilled water. The third-stage larvae (L3 were obtained after eight days. The crude extract of D. flagrans was effective in reducing the number of L3, with the following percentage reductions: group 1, 49.5%; group 2, 52.5%; group 3, 36.8%; and group 4, 57.7%; in relation to the control group (p > 0.05. The proteolytic activity of the crude extract was confirmed through the zymogram. The results from this study confirmed that the crude extract of the fungusD. flagrans could be used for controlling cyathostomin L3, and suggested that at least one protease of approximately 38 kDa was present.

  7. Interaction between copper oxide wire particles and Duddingtonia flagrans in lambs.

    Science.gov (United States)

    Burke, J M; Miller, J E; Larsen, M; Terrill, T H

    2005-11-25

    An experiment was completed to determine if copper oxide wire particles (COWP) had any effect on the activity of the nematode-trapping fungus Duddingtonia flagrans in growing lambs. COWP has been used recently as a dewormer in small ruminants because of nematode resistance to anthelmintics. D. flagrans has been used to control free-living stages of parasitic nematodes in livestock. Katahdin and Dorper lambs, 4 months of age, were administered no or 4 g COWP (n=24/dose) in early October 2003. Haemonchus contortus was the predominant gastrointestinal parasite during the trial, which was acquired naturally from pasture. Half the lambs from each COWP group were supplemented with corn/soybean meal with or without D. flagrans for 35 days. Fecal egg counts (FEC) and packed cell volume (PCV) were determined weekly between days 0 (day of COWP administration) and 35. Feces from lambs in each treatment group were pooled and three replicates per group were cultured for 14 days at room temperature. Larvae (L3) were identified and counted per gram of feces cultured. Treatment with COWP was effective in decreasing FEC, which remained low compared with FEC from lambs not treated with COWP. This led to an increase in PCV in these lambs (COWP x day, Pcopper on H. contortus, and the additional larval reducing effect exerted by the nematode destroying fungus D. flagrans, the expected result would be a much lower larval challenge on pasture when these two tools are used together in a sustainable control strategy.

  8. Duddingtonia flagrans: controle biológico de nematodeos de bovinos a campo Duddingtonia flagrans: biological control of cattle nematodes in the field

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    Marta Bañolas Jobim

    2008-11-01

    Full Text Available O controle biológico é um método para diminuir uma população pela utilização de antagonista natural. No presente estudo, testou-se a eficácia do fungo nematófago Duddingtonia flagrans no controle de nematódeos parasitos gastrintestinais de bovinos criados à campo no município de Júlio de Castilhos. Foram utilizados 20 bezerros, distribuídos igualmente em duas áreas formadas por pastagem nativa. O grupo A foi tratado com o fungo D. flagrans, cultivado em sorgo, numa concentração de 1x10(6clamidósporos kg-1 de peso animal, misturados em ração de manutenção, diariamente, durante oito meses. O grupo B serviu como controle e não recebeu fungo, apenas ração. Foram coletadas amostras para contagem de ovos por grama de fezes (OPG semanalmente. Mensalmente, foram realizadas coproculturas para identificar as espécies de larvas de nematódeos, a pesagem dos animais e a coleta de pasto para contagem das larvas na pastagem. Dados de temperatura e índice pluviométrico foram registrados diariamente. O OPG foi reduzido no grupo tratado, em média 56,8,% nos últimos três meses de experimento, variando entre 40,4 e 67,1% no grupo tratado (PBiological control is an alternative method to reduce parasite's population by the use of natural antagonist. In the present study, efficacy of nematophagous fungus Duddingtonia flagrans was tested to control gastrointestinal nematodes parasites of cattle livestock in the field. Twenty calves were used, distributed equally in two distinct plots formed by native pasture. Group A was treated with D. flagrans fungus, cultivated in sorghum (1x10(6 clamidospores kg-1 body weight mixed with maintenance ration, each day, during eight months. Group B served as a control and did not receive the fungus. Samples for faecal egg count (FEC, were collected each week. There were monthly counts in faecal cultures to identify the species of nematodes larvae, animals weight, blood collection to determine red cell

  9. The efficacy of two isolates of the nematode-trapping fungus Duddingtonia flagrans against Dictyocaulus viviparus larvae in faeces

    DEFF Research Database (Denmark)

    Fernandez, A.S.; Larsen, M.; Nansen, P.

    1999-01-01

    A series of experiments was carried out to examine the effects of two different isolates of the nematode-trapping fungus Duddingtonia flagrans to reduce the number of free-living larvae of the bovine lungworm, Dictyocaulus viviparus. A laboratory dose-titration assay showed that isolates CI3...

  10. Lack of negative effects of the biological control agent Duddingtonia flagrans on soil nematodes and other nematophagous fungi.

    Science.gov (United States)

    Saumell, C A; Fernández, A S; Echevarria, F; Gonçalves, I; Iglesias, L; Sagües, M F; Rodríguez, E M

    2016-11-01

    The possible environmental effects of the massive use of Duddingtonia flagrans for controlling sheep nematodes were evaluated in two regions. Non-supplemented faeces and faeces from sheep supplemented with D. flagrans were deposited three times on pasture plots and samples were collected 7 and 14 days post-deposition. Samples were cultured in agar-water (2%) with Panagrellus spp. to recover D. flagrans and other nematophagous fungi, and soil nematodes were extracted using Baermann funnels and counted. No significant differences in the populations of soil nematodes and fungi colonizing sheep faeces (P > 0.05) were observed between supplemented and non-supplemented groups, except in one sample. The topsoil in contact with the faeces was sampled 1-4 months post-deposition, revealing that, with one exception, D. flagrans did not persist in soil beyond 2 months post-deposition. Duddingtonia flagrans does not affect faecal colonization by other fungi and soil nematodes and, once deployed on pasture, does not survive for long periods in the environment.

  11. Effect of Duddingtonia flagrans against Ostertagia ostertagi in cattle grazing at different stocking rates

    DEFF Research Database (Denmark)

    Fernández, A.S.; Larsen, M.; Henningsen, E.

    1999-01-01

    divided into 3 comparable groups and allocated to 3 similar paddocks in each of the 2 trials. Two of the 3 groups received fungal material once per day during the initial 2 months, either at high dose (10(6) fungal spores/kg body weight) or low dose (5 x 10(5) or 2.5 x 10(5) fungal spores/kg body weight......The efficacy of an isolate of the nematophagous fungus Duddingtonia flagrans against gastrointestinal nematodes of cattle was examined at 2 dose levels on 2 permanent pastures, with high and low stocking rates, respectively. Thirty calves, experimentally infected with Ostertagia ostertagi, were...... seemed not to be very heavy, and a conclusive effect of the fungi at the dose-level used could not be detected....

  12. Duddingtonia flagrans formulated in rice bran in the control of Oesophagostomum spp. intestinal parasite of swine.

    Science.gov (United States)

    Facchini Rodrigues, João Victor; Braga, Fabio Ribeiro; Campos, Artur Kanadani; de Carvalho, Lorendane Millena; Araujo, Juliana Milani; Aguiar, Anderson Rocha; Ferraz, Carolina Magri; da Silveira, Wendeo Ferreira; Valadão, Marisa Caixeta; de Oliveira, Thais; de Freitas, Samuel Galvão; de Araújo, Jackson Victor

    2018-01-01

    Three experimental assays with Duddingtonia flagrans (isolated AC001) were carried out. The growth of the genus Duddingtonia present in formulation of rice bran, its predatory capability on Oesophagostomum spp. infective larvae (L 3 ) in petri dishes (assay 1), its action in faecal cultures with eggs of that parasite (assay 2) and isolate's capability of predation after passing through gastrointestinal tract of swine (assay 3) was evaluated. At assay 3, feces were collected at time intervals of 12, 24, 36, 48, and 60 h after feed animals with the formulation. Assays 1 and 2 showed a statistical difference (p  0.05). The results demonstrate that the fungal isolate AC001 formulated in rice bran can prey on L 3 of Oesophagostomum spp., in vitro and after passing through the gastrointestinal tract, without loss of viability. This isolate may be an alternative in the control of Oesophagostomum spp. in swine. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Extracellular biosynthesis of silver nanoparticles using the cell-free filtrate of nematophagous fungus Duddingtonia flagrans

    Directory of Open Access Journals (Sweden)

    Costa Silva LP

    2017-08-01

    Full Text Available Laryssa Pinheiro Costa Silva,1 Jairo Pinto Oliveira,2 Wanderson Juvencio Keijok,2 André Romero da Silva,3 Anderson Rocha Aguiar,1 Marco Cesar Cunegundes Guimarães,2 Carolina Magri Ferraz,1 Jackson Victor Araújo,4 Fernando Luiz Tobias,5 Fábio Ribeiro Braga1 1Department of Parasitology, University Vila Velha, Vila Velha, Espírito Santo, Brazil; 2Morphology Department, Federal University of Espírito Santo, Vitória, Espírito Santo, Brazil; 3Federal Institute of Education, Science and Technology of Espírito Santo, Aracruz, Espírito Santo, Brazil; 4Department of Veterinary Medicine, Federal University of Viçosa, Viçosa, Minas Gerais, Brazil; 5Department of Microbiology, University Vila Velha, Vila Velha, Espírito Santo, Brazil Abstract: The biosynthesis of metallic nanoparticles (NPs using biological systems such as fungi has evolved to become an important area of nanobiotechnology. Herein, we report for the first time the extracellular synthesis of highly stable silver NPs (AgNPs using the nematophagous fungus Duddingtonia flagrans (AC001. The fungal cell-free filtrate was analyzed by the Bradford method and 3,5-dinitrosalicylic acid assay and used to synthesize the AgNPs in the presence of a 1 mM AgNO3 solution. They have been characterized by UV–Vis spectroscopy, X-ray diffraction, transmission electron microscopy, dynamic light scattering, Zeta potential measurements, Fourier-transform infrared, and Raman spectroscopes. UV–Vis spectroscopy confirmed bioreduction, while X-ray diffractometry established the crystalline nature of the AgNPs. Dynamic light scattering and transmission electron microscopy images showed approximately 11, 38 nm monodisperse and quasispherical AgNPs. Zeta potential analysis was able to show a considerable stability of AgNPs. The N–H stretches in Fourier-transform infrared spectroscopy indicate the presence of protein molecules. The Raman bands suggest that chitinase was involved in the growth and

  14. Proteolytic action of the crude extract of Duddingtonia flagrans on cyathostomins (Nematoda: Cyathostominae in coprocultures Ação proteolítica do extrato bruto de Duddingtonia flagrans sobre ciatostomíneos (Nematoda: Cyathostominae em coproculturas

    Directory of Open Access Journals (Sweden)

    Fabio Ribeiro Braga

    2013-03-01

    Full Text Available The objective of this study was to examine the action of the crude extract of Duddingtonia flagrans (isolates AC001 and CG722 on infective larvae (L3 of cyathostomins in coprocultures and to confirm its proteolytic activity by means of a zymogram. The following groups were formed in coprocultures: Group 1: 10 mL of crude extract of D. flagrans (AC001; group 2: 10 mL of crude extract of AC001 with 10 mM of Ca2+; group 3: 10 mL of crude extract of D. flagrans (CG722; group 4: 10 mL of crude extract of CG722 with 10 mM of Ca2+; and group 5: control group (distilled water. The third-stage larvae (L3 were obtained after eight days. The crude extract of D. flagrans was effective in reducing the number of L3, with the following percentage reductions: group 1, 49.5%; group 2, 52.5%; group 3, 36.8%; and group 4, 57.7%; in relation to the control group (p > 0.05. The proteolytic activity of the crude extract was confirmed through the zymogram. The results from this study confirmed that the crude extract of the fungus D. flagrans could be used for controlling cyathostomin L3, and suggested that at least one protease of approximately 38 kDa was present.O objetivo deste trabalho foi estudar a ação do extrato bruto de Duddingtonia flagrans (isolados AC001 e CG722 sobre larvas infectantes (L3 de ciatostomíneos em coproculturas e confirmar a sua atividade proteolítica por meio de um zimograma. Foram formados os seguintes grupos em coproculturas: grupo 1: 10 mL de extrato bruto de D. flagrans (AC001; grupo 2: 10 mL de extrato bruto de AC001 com íons Ca2+ 10 Mm; grupo 3: 10 mL de extrato bruto de D. flagrans (CG722; grupo 4: 10 mL de extrato bruto de CG722 com íons Ca2+ 10 Mm; e grupo 5 como controle (água destilada, obtendo-se as L3 ao final de 8 dias. O extrato bruto de D. flagrans foi eficiente na redução do número de L3 com os seguintes percentuais de redução: grupo 1 (49,5%; grupo 2 (52,5%; grupo 3 (36,8% e grupo 4 (57,7% em relação ao grupo

  15. Activity of the nematophagous fungi Pochonia chlamydosporia, Duddingtonia flagrans and Monacrosporium thaumasium on egg capsules of Dipylidium caninum.

    Science.gov (United States)

    Araujo, Juliana Milani; de Araújo, Jackson Victor; Braga, Fabio Ribeiro; Carvalho, Rogério Oliva; Ferreira, Sebastião Rodrigo

    2009-12-03

    Nematophagous fungi are potential biological control agents of helminths. The in vitro ovicidal effect of four isolates of the nematophagous fungi Pochonia chlamydosporia (VC1 and VC4), Duddingtonia flagrans (AC001) and Monacrosporium thaumasium (NF34) was evaluated on egg capsules of Dipylidium caninum, a cestode parasite of dogs, cats and humans. One thousand egg capsules of D. caninum were plated on 2% water-agar with the grown isolates and control without fungus. The ovicidal activity of these fungi was evaluated 5, 10 and 15 days after incubation. Only P. chlamydosporia showed ovicidal activity (pcaninum egg capsules, of 19.6% (VC1) and 20% (VC4) on the 5th day; 44.2% (VC1) and 31.5% (VC4) on the 10th day; and 49.2% (VC1) and 41.9% (VC4) on the 15th day. D. flagrans and M. thaumasium caused no morphological damage to egg capsules. The results demonstrated that P. chlamydosporia was in vitro effective against capsules and eggs of D. caninum, and can be considered as a potential biological control agent for this helminth.

  16. Biotic and abiotic factors influencing growth rate and production of traps by the nematode-trapping fungus Duddingtonia flagrans when induced by Cooperia oncophora larvae

    DEFF Research Database (Denmark)

    Grønvold, J.; Wolstrup, J.; Nansen, P.

    1999-01-01

    A series of experiments on corn meal agar was carried out to evaluate the efficacy of the nematode-trapping fungus Duddingtonia flagrans in different abiotic and biotic conditions which occur in cow pats. Above a concentration of 50 parasitic larvae (L-3) cm(-2) the fungus produced a maximum...... of between 500 and 600 nets cm(-2) at 20 degrees C in 2 days on the surface of corn meal agar. There were no differences in the trap-producing capacity of three strains of D. flagrans (CIII4, CI3 and Trol A). On agar at 30 degrees and 20 degrees C, the fungus responded to Coaperia oncophora L-3 very quickly...... anaerobically. On agar, D. flagrans (CI3) did not produce trapping nets in an anaerobic atmosphere. Moreover, C. oncophora L-3 stopped migration under anaerobic conditions. When the fungal cultures were transferred to a normal aerobic atmosphere, after 1 and 2 weeks under anaerobic conditions, the C. oncophora...

  17. Optimizing protease production from an isolate of the nematophagous fungus Duddingtonia flagrans using response surface methodology and its larvicidal activity on horse cyathostomins.

    Science.gov (United States)

    Braga, F R; Araújo, J V; Soares, F E F; Araujo, J M; Genier, H L A; Silva, A R; Carvalho, R O; Queiroz, J H; Ferreira, S R

    2011-06-01

    Protease production from Duddingtonia flagrans (isolate AC001) was optimized and the larvicidal activity of the enzymatic extract was evaluated on infective horse cyathostomin larvae (L3). Duddingtonia flagrans was grown in liquid medium with eight different variables: glucose, casein, bibasic potassium phosphate (K2HPO4), magnesium sulphate (MgSO4), zinc sulphate (ZnSO4), ferrous sulphate (FeSO4), copper sulphate (CuSO4) and temperature. The Plackett-Burman analysis showed a significant influence of MgSO4, CuSO4 and casein (P < 0.05) on protease production by D. flagrans in liquid medium. Central composite design indicated that the highest proteolytic activity was 39.56 U/ml as a function of the concentrations of casein (18.409 g/l), MgSO4 (0.10 g/l) and CuSO4 (0.50 mg/l). A significant difference (P < 0.01) was found for the larval number between the treated and control groups at the end of the experiment. A reduction of 95.46% in the number of free-living larvae was found in the treated group compared with the control. The results of this study suggest that protease production by D. flagrans (AC001) in liquid medium was optimized by MgSO4, CuSO4 and casein, showing that the optimized enzymatic extract exerted larvicidal activity on cyathostomins and therefore may contribute to large-scale industrial production.

  18. Efficacy of the nematode-trapping fungus Duddingtonia flagrans against three species of gastro-intestinal nematodes in laboratory faecal cultures from sheep and goats.

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    Waghorn, T S; Leathwick, D M; Chen, L-Y; Skipp, R A

    2003-12-30

    The ability of the nematode-killing fungus Duddingtonia flagrans to reduce number of infective larvae of three species of gastro-intestinal parasitic nematodes developing in dung was investigated in both goats and sheep. Groups of lambs and kids (12-20 weeks old) were given mono-specific infections of Haemonchus contortus, Ostertagia (Teladorsagia) circumcincta or Trichostrongylus colubriformis. Following patency of the infections (t1) faecal samples were collected for determination of faecal nematode egg count (FEC) and culture of parasite larvae. Groups of animals were then dosed on 2 consecutive days with one of the two dose rates of the fungus (250,000 or 500,000 spores/kg liveweight). One (t2) and 5 (t3) days after the second dose of fungus samples were again collected for FEC and culture. The number of larvae recovered from the faecal cultures at t1 and t3 were used as controls to assess the efficacy of the experimental treatment at t2. Average efficacy was 78% with group means ranging from 40 to 93%. Dose rate of fungus appeared to influence efficacy against O. circumcincta but not against H. contortus or T. colubriformis. Overall, there were no differences in the efficacy of the fungus against any of the parasite species or in either host animal. The results of this trial indicate the potential use of this fungus as a broad spectrum anti-parasite agent for use in both goats and sheep.

  19. [Viability of nematophagous fungi Arthrobotrys robusta, Duddingtonia flagrans and Monacrosporium thaumasium after sporulation in different culture media].

    Science.gov (United States)

    Maciel, Alessandro S; de Araujo, Jackson V; Campos, Artur K

    2006-01-01

    Due to the shortage of studies that indicate the culture mediums that optimize the sporulation of namatophagous fungi for use in researche, the sporulation of the fungal isolates A. robusta (I31), D. flagrans (CG768) and M. thaumasium (NF34A) was evaluated in laboratorial conditions for 10 days in the means water-agar 2% (WA 2%), potato-dextrose-agar 2% (PDA 2%), corn-meal-agar 2% (CMA 2%) and yeast-phosphate-sulphate-sucrose-agar (YPSSA). The largest conidia production (P BDA 2% while in the isolates I31 and NF34A produced larger conidia number in YPSSA (P 0.05).

  20. Capability of the nematode-trapping fungus Duddingtonia flagrans to reduce infective larvae of gastrointestinal nematodes in goat feces in the southeastern United States: dose titration and dose time interval studies.

    Science.gov (United States)

    Terrill, T H; Larsen, M; Samples, O; Husted, S; Miller, J E; Kaplan, R M; Gelaye, S

    2004-04-15

    Infection with gastrointestinal nematodes, particularly Haemonchus contortus, is a major constraint to goat production in the southeastern United States. Non-anthelmintic control alternatives are needed due to increasing resistance of these nematodes to available anthelmintics. Two studies were completed in Central Georgia in August 1999, and April-May 2000, using Spanish does naturally infected with Haemonchus contortus, Trichostongylus colubriformis, and Cooperia spp. to evaluate effectiveness of nematode-trapping fungi as a biological control agent. In the first experiment, five levels of Duddingtonia flagrans spores were mixed with a complete diet and fed once daily to the does (three per treatment) in metabolism crates. The treatment concentrations were (1) 5 x 10(5), (2) 2.5 x 10(5), (3) 10(5), and (4) 5 x 10(4) spores per kilogram body weight (BW), and (5) no spores. Fungal spores were fed for the first 7 days of the 14-day trial, and fecal samples were collected daily from individual animals for analysis of fecal egg count and establishment of fecal cultures. Efficacy of the fungus at reducing development of infective larvae (L3) in the fecal cultures was evaluated. The mean reduction in L3 from day 2 of the treatment period until the day after treatment stopped (days 2-8) was 93.6, 80.2, 84.1, and 60.8% for animals given the highest to lowest spore doses, respectively. Within 3-6 days after termination of fungal spore feedings, reduction in L3 development was no longer apparent in any of the treated animals. In a second experiment, effectiveness of 2.5 x 10(5) spores of D. flagrans per kilogram BW fed to does every day, every second day, and every third day was evaluated. Reduction in L3 development by daily feeding was less in the second experiment than in the first experiment. Daily fungal spore feeding provided more consistent larval reduction than intermittant feeding (every second or third day). When fed daily under controlled conditions, D. flagrans

  1. Methods for assessing Phytophthora ramorum chlamydospore germination

    Science.gov (United States)

    Joyce Eberhart; Elilzabeth Stamm; Jennifer Parke

    2013-01-01

    Germination of chlamydospores is difficult to accurately assess when chlamydospores are attached to remnants of supporting hyphae. We developed two approaches for closely observing and rigorously quantifying the frequency of chlamydospore germination in vitro. The plate marking and scanning method was useful for quantifying germination of large...

  2. Germination of Phytophthora ramorum chlamydospores: a comparison of separation method and chlamydospore age

    Science.gov (United States)

    Justin P. Shaffer; Jennifer L. Parke

    2013-01-01

    Phytophthora ramorum characteristically produces large amounts of chlamydospores in vitro, but the role of these propagules in the disease cycle remains unclear. Germination is difficult to observe and quantify if chlamydospores are not free of mycelium, and the low frequency of germination commonly reported suggests that...

  3. The maturation and germination of Phytophthora ramorum Chlamydospores

    Science.gov (United States)

    Aaron L. Smith; Everett M. Hansen

    2008-01-01

    Chlamydospores are a distinctive feature of Phytophthora ramorum. They are formed quickly in agar, and within colonized leaves. We followed their development and maturation in vitro and in vivo, and studied conditions affecting their germination. Cell walls of mature P. ramorum chlamydospores...

  4. Purification and germination of Candida albicans and Candida dubliniensis chlamydospores cultured in liquid media

    OpenAIRE

    Citiulo, Francesco; Moran, Gary; COLEMAN, DAVID; SULLIVAN, DEREK

    2009-01-01

    PUBLISHED Candida albicans and Candida dubliniensis are the only Candida species that have been observed to produce chlamydospores. The function of these large, thick-walled cells is currently unknown. In this report we describe the production and purification of chlamydospores from these species in defined liquid media. Staining with the fluorescent dye FUN-1 indicated that chlamydospores are metabolically active cells, but that metabolic activity is undetectable in chlamydospores that...

  5. Purification and germination of Candida albicans and Candida dubliniensis chlamydospores cultured in liquid media.

    LENUS (Irish Health Repository)

    Citiulo, Francesco

    2009-10-01

    Candida albicans and Candida dubliniensis are the only Candida sp. that have been observed to produce chlamydospores. The function of these large, thick-walled cells is currently unknown. In this report, we describe the production and purification of chlamydospores from these species in defined liquid media. Staining with the fluorescent dye FUN-1 indicated that chlamydospores are metabolically active cells, but that metabolic activity is undetectable in chlamydospores that are >30 days old. However, 5-15-day-old chlamydospores could be induced to produce daughter chlamydospores, blastospores, pseudohyphae and true hyphae depending on the incubation conditions used. Chlamydospores that were preinduced to germinate were also observed to escape from murine macrophages following phagocytosis, suggesting that these structures may be viable in vivo. Mycelium-attached and purified chlamydospores rapidly lost their viability in water and when subjected to dry stress, suggesting that they are unlikely to act as long-term storage structures. Instead, our data suggest that chlamydospores represent an alternative specialized form of growth by C. albicans and C. dubliniensis.

  6. Incubation at room temperature may be an independent factor that induces chlamydospore production in Candida dubliniensis.

    Science.gov (United States)

    Sancak, Banu; Colakoglu, Sule; Acikgoz, Ziya Cibali; Arikan, Sevtap

    2005-08-01

    Production of chlamydospores is one of the phenotypic features used to differentiate Candida albicans and Candida dubliniensis. C. albicans produces few chlamydospores on only cornmeal/rice-Tween agar at room temperature, whereas C. dubliniensis produces abundant chlamydospores at this temperature both on cornmeal agar and some other commonly used media. We tried to determine whether the room temperature is the main factor that induces chlamydospore production of C. dubliniensis, regardless of the medium used. For this purpose, 100 C. albicans and 24 C. dubliniensis isolates were tested for chlamydospore production at room temperature and at 37 degrees C on some routinely used media, including eosin-methylene blue agar (EMB), nutrient agar (NA), nutrient broth (NB), and also on an investigational medium, phenol red agar (PR). At 37 degrees C, none of the isolates produced chlamydospores on any of the tested media. At 26 degrees C, all C. dubliniensis isolates produced abundant chlamydospores and pseudohyphae after 24-48 h on all tested media. At this incubation temperature, all C. albicans isolates failed to produce chlamydospores and pseudohyphae on EMB, NA, and NB, whereas 2 of the C. albicans isolates produced a few chlamydospores on PR. We also observed that all C. dubliniensis isolates tested on EMB and PR produced rough colonies with a hyphal fringe around the colonies, whereas none of the C. albicans isolates showed this property. In conclusion, incubation at 26 degrees C may play the key role for production of abundant chlamydospores and pseudohyphae by C. dubliniensis. Comprehensive molecular studies are needed to clarify the genetic basis of this observation. Using EMB and PR may be an inexpensive, a time-saving, and a simple way of presumptive identification of C. dubliniensis based on chlamydospore formation and colony morphology.

  7. Induction of chlamydospore formation in fusarium by cyclic lipopeptide antibiotics from Bacillus subtilis C2.

    Science.gov (United States)

    Li, Lei; Ma, Mingchuan; Huang, Rong; Qu, Qing; Li, Guohong; Zhou, Jinwei; Zhang, Keqin; Lu, Kaiping; Niu, Xuemei; Luo, Jun

    2012-08-01

    The culture filtrate of Bacillus subtilis strain C2 showed strong activity against the pathogenic fungus Fusarium solani f. sp. radicicola. A partially purified fraction (PPF) from the extract induced chlamydospore formation in Fusarium. Reverse-phase high performance liquid chromatography yielded 8 different fractions, six of which had chlamydospore-inducing activity. Mass spectrometry and nuclear magnetic resonance analyses identified the main active constituent as C(17) fengycin A (FA17), a cyclic lipopeptide. The effect of FA17 on morphology and physiology of two Fusarium species was dependent on the lipopeptide concentration. When challenged with FA17 at concentrations (0.5, 8, 64 μg ml(-1)) below the minimum inhibitory concentration (MIC) (128 μg ml(-1)), two species of Fusarium formed chlamydospores from hyphae, germ tubes, or inside the conidia within 2 days. At concentrations close to the MIC, FA17 caused Fusarium to form sparse and swollen hyphae or lysed conidia. The other five fractions were identified as fengycin A homologues. The homologues could also induce chlamydospore-like structures in 17 species of filamentous fungi including some specimens that do not normally produce chlamydospores, according to their taxonomic descriptions. Like other chlamydospores, these structures contained nuclei and lipid bodies as revealed by DAPI and Nile Red staining, and could germinate. This is the first study to demonstrate that under laboratory conditions fengycin, an antifungal lipopeptide produced by B. subtilis, can induce chlamydospore formation in Fusarium and chlamydospore-like structures in many filamentous fungi.

  8. Global Transcriptome Sequencing Identifies Chlamydospore Specific Markers in Candida albicans and Candida dubliniensis

    LENUS (Irish Health Repository)

    Palige, Katja

    2013-04-15

    Candida albicans and Candida dubliniensis are pathogenic fungi that are highly related but differ in virulence and in some phenotypic traits. During in vitro growth on certain nutrient-poor media, C. albicans and C. dubliniensis are the only yeast species which are able to produce chlamydospores, large thick-walled cells of unknown function. Interestingly, only C. dubliniensis forms pseudohyphae with abundant chlamydospores when grown on Staib medium, while C. albicans grows exclusively as a budding yeast. In order to further our understanding of chlamydospore development and assembly, we compared the global transcriptional profile of both species during growth in liquid Staib medium by RNA sequencing. We also included a C. albicans mutant in our study which lacks the morphogenetic transcriptional repressor Nrg1. This strain, which is characterized by its constitutive pseudohyphal growth, specifically produces masses of chlamydospores in Staib medium, similar to C. dubliniensis. This comparative approach identified a set of putatively chlamydospore-related genes. Two of the homologous C. albicans and C. dubliniensis genes (CSP1 and CSP2) which were most strongly upregulated during chlamydospore development were analysed in more detail. By use of the green fluorescent protein as a reporter, the encoded putative cell wall related proteins were found to exclusively localize to C. albicans and C. dubliniensis chlamydospores. Our findings uncover the first chlamydospore specific markers in Candida species and provide novel insights in the complex morphogenetic development of these important fungal pathogens.

  9. Chlamydospore production and germ-tube formation by auxotrophs of Candida albicans.

    Science.gov (United States)

    Balish, E

    1973-04-01

    A prototrophic strain and 21 auxotrophic strains of Candida albicans were assessed for their capacity to produce chlamydospores and germ tubes. All of the mutants were able to produce germ-tubes in human serum but only two mutants produced them in defined medium with L-alpha-amino-n-butyric acid as the sole source of nitrogen. Most auxotrophs were not able to produce chlamydospores on corn meal agar with 1% Tween 80, but they could be induced to do so if the medium was supplemented with their growth requirement(s). Although L-cysteine was able to support the growth of two methionine mutants, it did not support chlamydospore formation when added to corn meal agar with 1% Tween 80. Mutants of C. albicans that do not form chlamydospores could be incorrectly identified in laboratories that rely on chlamydospore formation for identification.

  10. Ovicidal activity of different concentrations of Pochonia chlamydosporia chlamydospores on Taenia taeniaeformis eggs.

    Science.gov (United States)

    Braga, F R; Silva, A R; Carvalho, R O; Araújo, J V; Pinto, P S A

    2011-03-01

    Three concentrations of chlamydospores of the nematophagous fungus Pochonia chlamydosporia (1000, 10,000 and 20,000 per Petri dish) were evaluated in vitro on Taenia taeniaeformis eggs. Chlamydospores at each concentration were cultured in two different media: 2% water-agar (2%WA) and 2% corn-meal-agar (2%CMA). Taenia taeniaeformis eggs were plated in each chlamydospore concentration in 2%WA and 2%CMA (treated groups) and without fungus (control group). Eggs were removed from each Petri dish at intervals of 7, 14 and 21 days and classified according to ovicidal activity (type 1, type 2 and type 3 effects). Plates containing 2%CMA showed the highest percentages for type 3 effect (81.3%) on the 21st day of observation. A difference (P < 0.01) between the media 2%WA and 2%CMA for type 1 effect was observed only at a concentration of 1000 chlamydospores on the 7th day. There were differences (P < 0.01) between 2%WA and 2%CMA on the 14th and 21st days, at the concentration of 20,000 chlamydospores, for type 1 and type 3 effects. Regression curves for type 3 effect in 2%WA and 2%CMA at the tested concentrations showed higher ovicidal activity with increasing chlamydospore concentrations. Results indicate that, at concentrations of 1000, 10,000 and 20,000 per Petri dish, chlamydospores of P. chlamydosporia effectively destroyed T. taeniaeformis eggs and can be considered a potential biological control agent for this cestode.

  11. Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation*

    Science.gov (United States)

    Li, Ya-qian; Song, Kai; Li, Ya-chai; Chen, Jie

    2016-01-01

    Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×108 spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×108 spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents. PMID:27487807

  12. Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation.

    Science.gov (United States)

    Li, Ya-Qian; Song, Kai; Li, Ya-Chai; Chen, Jie

    2016-08-01

    Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×10(8) spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×10(8) spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents.

  13. Rapid production of Candida albicans chlamydospores in liquid media under various incubation conditions.

    Science.gov (United States)

    Alicia, Zavalza-Stiker; Blanca, Ortiz-Saldivar; Mariana, García-Hernández; Magdalena, Castillo-Casanova; Alexandro, Bonifaz

    2006-01-01

    The production of chlamydospores is a diagnostic tool used to identify Candida albicans; these structures also represent a model for morphogenetic research. The time required to produce them with standard methods is 48-72 hours in rice meal agar and tensoactive agents. This time can be shorted using liquid media such as cornmeal broth (CMB) and dairy supplements. Five media were tested: CMB plus 1% Tween-80, CMB plus 5% milk, CMB plus 5% milk serum, milk serum, and milk serum plus 1% Tween-80, under different incubation conditions: at 28 degrees C and 37 degrees C in a metabolic bath stirring at 150 rpm, and at 28 degrees C in a culture stove. The reading time points were established at 8 and 16 hours. The best results were obtained at 16 hours with CMB plus 5% milk under incubation at 28 degrees C and stirring at 150 rpm. The next most efficient methods were CMB plus 5% milk serum and CMB plus 1% Tween-80, under the same incubation conditions. The other media were ineffective in producing chlamydospores. The absence of stirring at 28 degrees C prevented the formation of chlamydospores within the set time points, and incubation at 37 degrees C decreased their production. This paper reports that the time to form C. albicans chlamydospores can be reduced.

  14. Effects of 60 Cobalt ionizing radiation in morphology and metabolism of yeasts and Chlamydospore of Candida albicans

    International Nuclear Information System (INIS)

    Grillo, Michel R.F.; Demicheli, Marina C.; Andrade Junior, Heitor F.; Galiesteo Junior, Andres A.J.

    2015-01-01

    Candida albicans is a fungus responsible for 80-90% of fungal infections, as the symptoms are similar to those of systemic bacterial infections there is a difficulty for immediate diagnosis. These difficulties can lead to delays of antifungal therapy, which contributes to the high mortality rates associated with this infection. Resistance structures referred to as chlamydospores are very common in the pathogen, representing different cell types that form in response to certain genetic or environmental conditions. Recently, various antifungal agents and new therapeutic strategies have come into use, allowing the fungus to acquire a resistance to the drugs. The use of ionizing radiation has been widely employed for the production of immunogens against various parasites. In this work, we evaluate the effects of gamma radiation ( 60 Co) in yeast and chlamydospore of C. albicans with doses ranging from 320 to 10.240 Gy with Cobalt 60. Subsequently the samples were plated and after seven days, the colony forming units (CFU) told. The viability of irradiated cells were evaluated using the Janus green dye. A dose of 6000 Gy was considered ideal for the mitigation of chlamydospore and yeast. The dimorphic change mechanisms of both fungal structures were not harmed. The viability of chlamydospores remained above 70% while the yeast viability remained above 85%. By transmission electron microscopy and fluorescence microscopy may be noted cytoplasmic changes, defects in the cell wall, mitochondria, and the presence of partially preserved vesicles of both morphological forms of C. albicans. Irradiation both chlamydospore as C. albicans yeast allows the suppression of their reproduction, opening the possibility of their use in future candidate immunogens. (author)

  15. Effects of 60 Cobalt ionizing radiation in morphology and metabolism of yeasts and Chlamydospore of Candida albicans

    Energy Technology Data Exchange (ETDEWEB)

    Grillo, Michel R.F.; Demicheli, Marina C.; Andrade Junior, Heitor F.; Galiesteo Junior, Andres A.J., E-mail: galisteo@usp.br [Universidade de Sao Paulo (IMTSP/USP), Sao Paulo, SP (Brazil). Instituto de Medicina Tropical. Lab. de Protozoologia; Takakura, Cleusa F.H. [Universidade de Sao Paulo (FM/USP), Sao Paulo, SP (Brazil). Departamento de Patologia de Molestias Transmissiveis. Lab. de Patologia; Negro, Gilda M.B. del [Universidade de Sao Paulo (HCFM/USP/IMTSP/LIM-53), Sao Paulo, SP (Brazil). Hospital das Clinicas. Lab. de Micologia; Nascimento, Nanci do [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2015-07-01

    Candida albicans is a fungus responsible for 80-90% of fungal infections, as the symptoms are similar to those of systemic bacterial infections there is a difficulty for immediate diagnosis. These difficulties can lead to delays of antifungal therapy, which contributes to the high mortality rates associated with this infection. Resistance structures referred to as chlamydospores are very common in the pathogen, representing different cell types that form in response to certain genetic or environmental conditions. Recently, various antifungal agents and new therapeutic strategies have come into use, allowing the fungus to acquire a resistance to the drugs. The use of ionizing radiation has been widely employed for the production of immunogens against various parasites. In this work, we evaluate the effects of gamma radiation ({sup 60}Co) in yeast and chlamydospore of C. albicans with doses ranging from 320 to 10.240 Gy with Cobalt 60. Subsequently the samples were plated and after seven days, the colony forming units (CFU) told. The viability of irradiated cells were evaluated using the Janus green dye. A dose of 6000 Gy was considered ideal for the mitigation of chlamydospore and yeast. The dimorphic change mechanisms of both fungal structures were not harmed. The viability of chlamydospores remained above 70% while the yeast viability remained above 85%. By transmission electron microscopy and fluorescence microscopy may be noted cytoplasmic changes, defects in the cell wall, mitochondria, and the presence of partially preserved vesicles of both morphological forms of C. albicans. Irradiation both chlamydospore as C. albicans yeast allows the suppression of their reproduction, opening the possibility of their use in future candidate immunogens. (author)

  16. Inoculating chlamydospores of Trichoderma asperellum SM-12F1 changes arsenic availability and enzyme activity in soils and improves water spinach growth.

    Science.gov (United States)

    Su, Shiming; Zeng, Xibai; Bai, Lingyu; Williams, Paul N; Wang, Yanan; Zhang, Lili; Wu, Cuixia

    2017-05-01

    Arsenic (As)-contaminated agricultural soils threaten crop yields and pose a human health risk. Augmentation of exogenous microorganisms exhibiting plant-growth promoting and As speciation changing shows potential to improve crop growth and change soil As availability. Trichoderma asperellum SM-12F1 exhibiting both traits was developed into chlamydospores to improve its persistence in contaminated soils. After inoculation, As availability and enzyme activity in two types of soils and the growth as well as As uptake of water spinach (Ipomoea aquatic Forsk.) were investigated. The results indicated that inoculation significantly improved water spinach growth in both soils. Inoculating chlamydospores at 5% significantly increased As concentration (139%), bioconcentration factor (150%), and translocation factor (150%) in water spinach grown in Chenzhou (CZ) soils, while no significant change for these in Shimen (SM) soils. Inoculating chlamydospores at 5% caused a significant increase (16%) of available As content in CZ soils, while a significant decrease (13%) in SM soils. Inoculation significantly caused As methylation in both soils, while significant As reduction merely observed in CZ soils. The differential changes in available As contents in both soils were attributed to the soil pH, As fractionations and speciation characteristics. Furthermore, Inoculating chlamydospores at 5% significantly improved the activities of β-glucosidase (155%), chitinase (211%), and phosphatase (108%) in SM soils, while significant decreases in β-glucosidase (81%), phosphatase (54%), aminopeptidase (60%), and catalase (67%) in CZ soils. Bioaugmentation and As availability change were responsible for this result. These observations will be helpful for the application of fungal chlamydospores in the future bioremediation. Copyright © 2017. Published by Elsevier Ltd.

  17. Produção de clamidósporos de Pochonia chlamydosporia em diferentes substratos Production of chlamydospores of Pochonia chlamydosporia in different substrates

    Directory of Open Access Journals (Sweden)

    Rosangela Dallemole-Giaretta

    2011-04-01

    no forno de micro-ondas. O melhor meio de cultivo de P. chlamydosporia para a produção de clamidósporos foi o substrato contendo grãos de arroz.Chlamydospores are survival structures of the nematophagous fungus Pochonia chlamydosporia. The objective of this study was to evaluate different substrates, different contents of water and kinds of inoculum for the production of Pochonia chlamydosporia chlamydospores. The substrates evaluated were: milled maize, rice grains and coffee husk; all of them were inoculated with disks of culture, concentrated liquid media or diluted liquid media (1:40 colonized by P. chlamydosporia. Besides, the substrates milled maize, supplemented or not with potato broth, and inoculated with fungus disks or aqueous suspension were evaluated. Milled maize and coffee husk were the best substrates for chlamydospores production. The best inoculum kinds were disks of culture and diluted liquid media (1:40 colonized by P. chlamydosporia. The supplementation of milled maize with potato broth did not improve the production of chlamydospores, and the best inoculum form was disks of fungus. Moreover, it was also studied the substrates rice grains, coffee husk and coconut fibers with different rates of water; the substrate coffee husk moistened by different periods; the supplementation of coffee husk with rice flour or dextrose; and the substrate grains of rice sterilized in microwave oven with different rates of water. The substrate grains of rice, in all of the amounts of water tested showed increase in production of chlamydospores. There was no difference in the chlamydospores production of the coffee husk substrate, when moistened by different periods of time and only when it was supplemented with rice flour it showed higher mean of clamydospores per gram of substrate. All treatments evaluated showed higher production of chlamydospores in the substrate grains of rice treated in the microwave oven. The best substrate for the production of

  18. [Effects of pesticides and plant bio-stimulants on the germination of chlamydospores and in vitro development of the nematophagous fungus Pochonia chlamydosporia].

    Science.gov (United States)

    Ceiro, Wilson G; Arévalo, Jersys; Hidalgo-Díaz, Leopoldo

    2015-01-01

    The effects of pesticides and plant bio-stimulants used in protected vegetable production systems on the fungus Pochonia chlamydosporia are unknown. The effectiveness of P. chlamydosporia against Meloidogyne spp. could be affected by products used in protected vegetable production systems. Two in vitro assays were carried out to evaluate any potential effect that pesticides and bio-stimulants often used in these systems could have on the fungus. The effect on chlamydospore germination was evaluated in a first assay, and mycelia growth and sporulation in a second. With these results, the compatibility of each product with the fungus was determined. Chlamydospores germination was over 50% with the control, FitoMas E, Biobras-16 and Amidor. Lower results were observed with other products, with some of them even inhibiting germination completely. Fungal growth was potentiated by Biobras-16 to 106.23%, promoted up to 50-100% by the control, FitoMas E and Cuproflow, and was below 50% with the rest of the products.Cipermetrina, Benomilo, Zineb, Mitigan, Karate, FitoMas E and Amidor promoted fungal sporulation, which was below 50% with Cuproflow and completely inhibited by the other products. Fifty-four percent of the products evaluated were compatible with P. chlamydosporia, while 8% were toxic and 38%, very toxic. Cipermetrina, Karate, Amidor, Benomilo, Zineb, Mitigan and FitoMas E were compatible with P. chlamydosporia. If it is necessary to use any of the other products for integrated pest management in protected vegetable production systems, it is recommended to avoid direct contact with P. chlamydosporia. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  19. Growth rate and trapping efficacy of nematode-trapping fungi under constant and fluctuating temperatures

    DEFF Research Database (Denmark)

    Fernandez, A.S.; Larsen, M.; Wolstrup, J.

    1999-01-01

    The effect of temperature on radial growth and predatory activity of different isolates of nematode-trapping fungi was assessed. Four isolates of Duddingtonia flagrans and one isolate of Arthrobotrys oligospora were inoculated on petri dishes containing either cornmeal agar (CMA) or faecal agar...

  20. Mixed Production of Filamentous Fungal Spores for Preventing Soil-Transmitted Helminth Zoonoses: A Preliminary Analysis

    Directory of Open Access Journals (Sweden)

    M. S. Arias

    2013-01-01

    Full Text Available Helminth zoonoses are parasitic infections shared by humans and animals, being the soil-transmitted helminths (STHs mainly caused by roundworms (ascarids and hookworms. This study was aimed to assess the individual and/or mixed production of two helminth-antagonistic fungi, one ovicide (Mucor circinelloides and other predator (Duddingtonia flagrans. Fungi were grown both in Petri plates and in a submerged culture (composed by water, NaCl, Na2HPO4 · 12 H2O, and wheat (Triticum aestivum. A Fasciola hepatica recombinant protein (FhrAPS was incorporated to the cultures to improve fungal production. All the cultured plates showed fungal growth, without difference in the development of the fungi when grown alone or mixed. High counts of Mucor spores were produced in liquid media cultures, and no significant differences were achieved regarding single or mixed cultures, or the incorporation of the FhrAPS. A significantly higher production of Duddingtonia spores after the incorporation of the FhrAPS was observed. When analyzing the parasiticide efficacy of the fungal mixture, viability of T. canis eggs reduced to 51%, and the numbers of third stage cyathostomin larvae reduced to 4%. It is concluded, the capability of a fungal mixture containing an ovicide (Mucor and a predator species (Duddingtonia for growing together in a submerged medium containing the FhrAPS offers a very interesting tool for preventing STHs.

  1. Predatory activity of Butlerius nematodes and nematophagous fungi against Haemonchus contortus infective larvae

    Directory of Open Access Journals (Sweden)

    Manoel Eduardo da Silva

    Full Text Available Abstract The purpose of this study was to evaluate the predatory activity of the nematode Butlerius spp. and fungal isolates of Duddingtonia flagrans, Clonostachys rosea, Arthrobotrys musiformis and Trichoderma esau against H. contortus infective larvae (L3 in grass pots. Forty-eight plastic gardening pots containing 140 g of sterile soil were used. Panicum spp. grass seeds (200 mg were sown into each pot and individually watered with 10 mL of tap water. Twelve days after seeding, the pots were randomly divided into 6 groups (n=8. Two thousand H. contortus infective larvae (L3 were added to each group. Additionally, the following treatments were established: Group 1 – 2000 Butlerius spp. larvae; group 2 – A. musiformis (1x107 conidia; group 3 – T. esau (1x107 conidia; group 4 – C. rosea (1x107 conidia, group 5 – D. flagrans (1x107conidia and Group 6 – no biological controller (control group. The larval population of H. contortus exposed to Butlerius spp. was reduced by 61.9%. Population reductions of 90.4, 66.7, 61.9 and 85.7% were recorded in the pots containing A. musiformis, T. esau, C. rosea and D. flagrans, respectively. The results of this study indicate that the predatory nematode Butlerius spp. and the assessed fungi display an important predatory activity can be considered suitable potential biological control agents.

  2. Screening of different sample types associated with sheep and cattle for the presence of nematophagous fungi in China.

    Science.gov (United States)

    Cai, Kui-Zheng; Liu, Jun-Lin; Liu, Wei; Wang, Bo-Bo; Xu, Qiang; Sun, Long-Jie; Chen, Ming-Yue; Zhao, Ming-Wang; Wu, Jia-Yan; Li, Xiao-Shan; Yang, Jing; Wei, Shuan; Chen, Chun-Rong; Ma, Zhong-Ren; Xu, Chun-Lan; Wang, Feng; Hu, Qian-Lin; Fang, Wen-Xiu; Zheng, Tian-Hui; Wang, Yue-Ying; Zhu, Wen-Long; Li, Dan; Li, Qing; Zhang, Chao; Cai, Bing; Wang, Fan; Yang, Zai-Yun; Liu, Yan-Qiu

    2016-03-01

    A total of 1502 samples, including feces of sheep (793) and cattle (348), pasture soil (118), dung compost (147) and barn soil (96), were examined between October 2012 and August 2014 to discover potential strains of nematophagous fungi for the biological control of livestock-parasitic nematodes. These samples were collected from 87 sites located in 48 counties of 20 provinces (autonomous regions/municipalities) of China. Fungi were identified down to a species level. Four hundred and seventy-seven isolates, which were distributed in 8 genera and 28 taxa, were identified as nematophagous fungi. Nematode-trapping fungi included 17 species and one unidentified species of Arthrobotrys, two of Dactylella, Drechslerella dactyloides, and Duddingtonia flagrans. Five identified species and two unidentified species of endoparasitic fungi were isolated. The predominant species from all regions were Arthrobotrys oligospora, followed by Arthrobotrys musiformis, Arthrobotrys (Monacrosporium) thaumasiun, and Arthrobotrys (Monacrosporium) microscaphoides. Species with adhesive networks were the most frequently isolated. Among the endoparasitic fungi, Podocrella harposporifera (Harposporium anguillulae) was the most common species, followed by Harposporium lilliputanum and Harposporium arcuatum. Based on Shannon diversity index, the diversity levels of nematophagous fungi were relatively higher in samples associated with cattle, barn soil, and subtropical monsoon climate zone. Three species isolated from this study, namely, Duddingtonia flagrans, Arthrobotrys salina (Monacrosporium salinum), and Arthrobotrys oligospora var. sarmatica, are newly recorded in China, and 20 species (including one unidentified species) are newly recorded in sheep and cattle barn soils worldwide. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Control of infective larvae of gastrointestinal nematodes in heifers using different isolates of nematophagous fungi Controle de larvas infectantes de nematóides gastrintestinais de novilhas por diferentes isolados dos fungos nematófagos

    Directory of Open Access Journals (Sweden)

    Manoel Eduardo da Silva

    Full Text Available The effect of different nematophagous fungi [Duddingtonia flagrans (AC001 and CG722 and Monacrosporium thaumasium (NF34] with regard to controlling infective larvae (L3 of nematodes after gastrointestinal transit in female cattle (3/4 Holstein × Zebu was evaluated. A total of 24 pubescent female cattle were used, weighing approximately 320 kg each one. There were three treatment groups, each contained six animals that received 150 g of pellets (0.2 g of mycelium, orally in a single dose, in a sodium alginate matrix containing mycelial mass of the fungus D. flagrans (AC001 or CG722 or M. thaumasium (NF34; and one control group (without fungi. Fecal samples were collected from the animals at intervals of 12, 15, 18, 21, 24, 48, and 72 hours. At the end of 17 days, the L3 not subjected to predation were recovered by means of the Baermann method. The fungal isolates tested were capable of destroying the L3 after gastrointestinal transit. It was observed that within 72 hours, the isolates AC001, CG722, and NF34 showed a higher predatory activity (81.2%, 97.3%, and 98.3%, respectively. The results justify the need for studies in the field, and over longer intervals, in order to observe the efficiency of the fungus D. flagrans, or even M. thaumasium, for environmental control over nematodes in naturally infected cattle.No presente estudo, foi avaliado o efeito de diferentes fungos nematófagos [Duddingtonia flagrans (AC001 e CG722 e Monacrosporium thaumasium (NF34] no controle de larvas infectantes (L3 de nematóides após o trânsito gastrointestinal em fêmeas bovinas (3/4 Holandês x Zebu. Um total de 24 fêmeas bovinas pubescentes foram utilizadas, pesando aproximadamente 320 kg cada. Foram utilizados três grupos de tratamento; cada um contendo seis animais que receberam por via oral de 150 g de péletes (0,2 g de micélio, em dose única, em uma matriz de alginato de sódio contendo massa micelial dos fungos D. flagrans (AC001 ou

  4. In vitro susceptibility of nematophagous fungi to antiparasitic drugs: interactions and implications for biological control

    Directory of Open Access Journals (Sweden)

    J. N. Vieira

    Full Text Available Abstract The fast anthelmintic resistance development has shown a limited efficiency in the control of animal’s endoparasitosis and has promoted research using alternative control methods. The use of chemicals in animal anthelmintic treatment, in association with nematophagous fungi used for biological control, is a strategy that has proven to be effective in reducing the nematode population density in farm animals. This study aims to verify the in vitro susceptibility of the nematophagous fungi Arthrobotrys oligospora, Duddingtonia flagrans and Paecilomyces lilacinus against the antiparasitic drugs albendazole, thiabendazole, ivermectin, levamisole and closantel by using the Minimum Inhibitory Concentration (MIC. MICs ranged between 4.0 and 0.031 µg/mL for albendazole, thiabendazole and ivermectin, between 0.937 and 0.117 µg/mL for levamisole, and between 0.625 and 0.034 µg/mL for closantel. The results showed that all antiparasitic drugs had an in vitro inhibitory effect on nematophagous fungi, which could compromise their action as agents of biological control. D. flagrans was the most susceptible species to all drugs.

  5. Coadministration of nematophagous fungi for biological control over gastrointestinal helminths in sheep in the semiarid region of northeastern Brazil.

    Science.gov (United States)

    Vilela, Vinícius Longo Ribeiro; Feitosa, Thais Ferreira; Braga, Fabio Ribeiro; Araújo, Jackson Victor de; Santos, Antonielson dos; Morais, Dayana Firmino de; Souto, Diego Vagner de Oliveira; Athayde, Ana Célia Rodrigues

    2016-05-15

    This study aimed to evaluate coadministration of Duddingtonia flagrans and Monacrosporium thaumasium in a sodium alginate matrix for controlling gastrointestinal helminths in young and adult sheep in the semiarid region of northeastern Brazil. An area of 1ha was divided into two paddocks, in which two experimental groups (fungus and control) were formed, each consisting of six adult females and ten young males. In each group, two subgroups were formed in accordance with the animal category (adult or young). In the fungus group, each animal received 3g of pellets containing 0.6g of fungal mycelium, with 0.3g of D. flagrans and 0.3g of M. thaumasium for each 10 kg of body weight, in their feed twice a week, for six months. In the control group, each animal received 3g of pellets without fungus for each 10 kg of body weight, in their feed twice a week, for six months, serving as a witness group. Reductions in numbers of eggs per gram of feces of 76% among the adult sheep in the fungus group and 83% among the young sheep in the fungus group were observed, in comparison with their respective control subgroups. The groups that received these fungi needed less salvage deworming and presented better packed cell volume percentages, better weight gain and lower levels of L3/kg dry matter in their paddock than the control groups. Thus, it was concluded that coadministration of D. flagrans and M. thaumasium was effective in controlling gastrointestinal helminths of adults and young sheep in the semiarid region of northeastern Brazil. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Nematophagous fungi from decomposing cattle faeces in Argentina.

    Science.gov (United States)

    Saumell, Carlos Alfredo; Fernández, Alicia Silvina; Fusé, Luis Alberto; Rodríguez, Manuela; Sagüés, María Federica; Iglesias, Lucía Emilia

    2015-01-01

    Biological control of gastrointestinal nematodes of ruminants by use of nematophagous fungi would become part of any livestock parasite integral control system. Identifying autochthonous species that could then be selected for mass production is an important phase in the practical use of biological control. To search for nematophagous fungi with potential use as biological control agents against gastrointestinal nematodes in Argentina. Decomposing cattle faeces sampled in different locations were incubated in water agar 2% with Panagrellus sp. The developed nematophagous fungi were transferred to new water agar 2% plates and then to corn meal agar plates in order to carry out their identification. Fungal diversity and richness were also assessed. Seventeen species from nine genera of nematophagous fungi were found. Twelve species were nematode-trapping fungi and three species plus two fungi identified to genus level corresponded to endoparasitic fungi. Arthrobotrys conoides, Arthrobotrys oligospora, Duddingtonia flagrans, Monacrosporium doedycoides, Arthrobotrys robusta and Drechmeria coniospora were the most frequently isolated species overall in the whole study (6.6%, 5.7%, 5.7%, 5.7%, 4.7% and 4.7%, respectively) although other species were more frequently recorded at local levels such as Arthrobotrys pyriformis (18.8%). Only A. conoides has been previously isolated from ruminant faecal samples in Argentina. Five nematode-trapping fungal species are mentioned for the first time in the Americas D. flagrans and A. conoides, both identified in the present study, are among the most promising ones as biological control agents against gastrointestinal nematodes of ruminants. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  7. Survival and chlamydospore production of Phytophthora ramorum in California bay laurel leaves

    Science.gov (United States)

    E. Fichtner; D. Rizzo; S. Lynch; D. Rizzo; G. Buckles; J. Parke

    2009-01-01

    Sudden oak death manifests as non-lethal foliar lesions on bay laurel (Umbellularia californica), which support sporulation and survival of Phytophthora ramorum in forest ecosystems. The pathogen survives the dry summers in a proportion of attached bay leaves, but the propagules responsible for survival are...

  8. Anaerobic soil disinfestation reduces survival and infectivity of Phytophthora nicotianae chlamydospores in pepper

    Science.gov (United States)

    Phytophthora nicotianae is the principal causal agent of root and crown rot disease of pepper plants in Extremadura (western Spain), a spring-summer crop in this region. Preplant soil treatment by anaerobic soil disinfestation (ASD) may effectively control plant pathogens in many crop production sys...

  9. Biological control of infective larvae of Ancylostoma spp. in beach sand.

    Science.gov (United States)

    De Mello, Ingrid Ney Kramer; Braga, Fabio R; Monteiro, Thalita S Avelar; Freitas, Leandro G; Araujo, Juliana M; Soares, Filippe E Freitas; Araújo, Jackson V

    2014-01-01

    Geohelminths are parasites that stand out for their prevalence and wide distribution, depending on the soil for their transmission. The aim of this work was to evaluate the predatory capacity of the fungal isolate of the genus Duddingtonia (CG768) on third stage larvae (L3) of Ancylostoma spp. in beach sand under laboratory conditions. In the assay A five treatment groups and 1 control group were formed. The treatment groups contained 5000, 10,000, 15,000, 20,000 or 25,000 chlamydospores of the fungal isolate and 1000 Ancylostoma spp. L3 in pots containing 30g of sand. The control group (without fungus) contained only 1000 Ancylostoma spp. L3 and distilled water in pots with 30g of sand. Evidence of predatory activity was observed at the end of 15 days, where we observed the following percentages of reduction of L3: Group 1 (4.5%); Group 2 (24.5%); Group 3 (59.2%); Group 4 (58.8%); Group 5 (63%). However, difference was noted (p<0.01) only at concentrations 15,000, 20,000 and 25,000 in relation to control group. In the assay B two groups were formed in Petri dishes of 9cm in diameter containing agar water 2% medium. In the treated group, each Petri dish contained 500 Ancylostoma spp. L3 and 5g of sand containing the isolate CG 768 at a concentration of 25,000 chlamydospores/g of sand, and the control group (without fungus) contained only 500 L3. At the end of 7 days the non-predation L3 of Petri dishes using the method of Baermann were recovered. Difference (p<0.01) between groups on reducing the average number of Ancylostoma spp. L3 (percent reduction of 84%) was observed. The results of this study confirm earlier work on the efficiency of the Duddingtonia genus in the control of Ancylostoma spp. infective larvae. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  10. St. Mary's Hospital, Shercock Road, Castleblayney, Monaghan.

    LENUS (Irish Health Repository)

    Palige, Katja

    2013-04-15

    Candida albicans and Candida dubliniensis are pathogenic fungi that are highly related but differ in virulence and in some phenotypic traits. During in vitro growth on certain nutrient-poor media, C. albicans and C. dubliniensis are the only yeast species which are able to produce chlamydospores, large thick-walled cells of unknown function. Interestingly, only C. dubliniensis forms pseudohyphae with abundant chlamydospores when grown on Staib medium, while C. albicans grows exclusively as a budding yeast. In order to further our understanding of chlamydospore development and assembly, we compared the global transcriptional profile of both species during growth in liquid Staib medium by RNA sequencing. We also included a C. albicans mutant in our study which lacks the morphogenetic transcriptional repressor Nrg1. This strain, which is characterized by its constitutive pseudohyphal growth, specifically produces masses of chlamydospores in Staib medium, similar to C. dubliniensis. This comparative approach identified a set of putatively chlamydospore-related genes. Two of the homologous C. albicans and C. dubliniensis genes (CSP1 and CSP2) which were most strongly upregulated during chlamydospore development were analysed in more detail. By use of the green fluorescent protein as a reporter, the encoded putative cell wall related proteins were found to exclusively localize to C. albicans and C. dubliniensis chlamydospores. Our findings uncover the first chlamydospore specific markers in Candida species and provide novel insights in the complex morphogenetic development of these important fungal pathogens.

  11. Discrimination and numerical analysis of human pathogenic ...

    African Journals Online (AJOL)

    SERVER

    2008-02-19

    Minitab Inc. Pennsylvania, USA). RESULTS. Every one of the isolates exhibited characteristic oval budding yeast cells, germ tube and clusters of blastos- pore and terminal chlamydospore on Sabouraud's. Dextrose Agar medium.

  12. <原著>Candida albicansの厚膜胞子形成培地の検討

    OpenAIRE

    鎌口,有秀/馬場,久衛/小松,始/野崎,善弘/越前,敏廣; カマグチ,アリヒデ/ババ,ヒサエ/コマツ,ハジメ/ノザキ,ヨシヒロ/エチゼン,トシヒロ; KAMAGUCHI,Arihide/BABA,Hisae/KOMATSU,Hazime/NOSAKI,Yoshihiro/ECHIZEN,Toshihiro

    1987-01-01

    The composition of a medium and condition to reduce chlamydospore forming time in Candida albicans were examined. The most suitable composition of a medium and condition in reducing chlamydospore forming time were that the medium contained 30% of corn meal agar and 2 mg per ml of N-acetyl-D-glucosamine with an initial pH of 8.0, and incubation temperature was 25℃. This was designated 0.3CMG agar medium in this paper. On this medium all tested strains of C. albicans containing clinically isola...

  13. Susceptibility to Phytophthora ramorum and inoculum production potential of some common eastern forest understory plant species

    Science.gov (United States)

    Paul W. Tooley; Marsha Browning

    2009-01-01

    Twenty-five plant species (21 genera, 14 families), which comprise a portion of the understory in forests of the Eastern United States, were evaluated for susceptibility to infection by Phytophthora ramorum. The degree to which P. ramorum is able to form sporangia and chlamydospores was also assessed on...

  14. Biotechnology of Aureobasidium pullulans: A phylogenetic perspective

    Science.gov (United States)

    Aureobasidium pullulans is a fungus historically included among the "black yeasts." Although many strains are predominantly yeast-like, the species is actually polymorphic, exhibiting complex forms ranging from blastic conidia and swollen cells to pseudophyphae, hyphae, and chlamydospores. A. pull...

  15. Experimental minimum threshold for Phytophthora cinnamomi root disease expression on Quercus suber

    Directory of Open Access Journals (Sweden)

    María Socorro SERRANO

    2015-12-01

    Full Text Available Quercus suber seedlings were potted in soils infested with increasing concentrations of Phytophthora cinnamomi chlamydospores and submitted to weekly flooding for 3 months to favour root infections. Increasing quantities of chlamydospores led to an exponential increase in their ability to germinate. Root symptoms (necrosis and/or absence of feeder roots were significantly more severe than those recorded in uninfested soil only for plants potted in soils infested with 61 cfu g-1 or more. Although generated using potting mix, this minimum threshold represents a tool for checking the potential infectivity of infested soils or to assess the effectiveness of some control methods to reduce soil inoculum. However, a low level of root infection was recorded even at 3 cfu g-1. Therefore, long-term disease risk may be present whenever the pathogen is detectable in oak forest soils.

  16. Hichrom candida agar for identification of candida species

    OpenAIRE

    Baradkar V; Mathur M; Kumar S

    2010-01-01

    Chromogenic media are frequently used in direct and rapid identification of yeasts because different Candida species produce unique colors on these media. We used 60 isolates of Candida species including 30 C. albicans, 10 C. parapsilosis, 11 C. glabrata, five C. tropicalis, and four C. dubliniensis, isolated from various clinical specimens, to evaluate the performance of HiChrome Candida agar. These strains had been identified by germ tube test, morphology on cornmeal agar, chlamydospore for...

  17. The development and structure of thick-walled, multicellular, aerial spores in Diheterospora chlamydosporia (=Verticillium chlamydosporium).

    Science.gov (United States)

    Cambell, W P; Griffiths, D A

    1975-07-01

    The aerial, thick-walled spores in Diheterospara chlamydosporia arose as terminal swellings on erect hyphae. Repeated septation of the continuously swelling spore resulted in a multicellular structure. Immediately after the onset of septation secondary wall material was laid down between the two-layered primary wall and the plasmalemma. The presence of secondary wall material indicates that the multicellular spore is a dictyochlamydospore and not an aleuriospore. The relationship between chlamydospores and aleuriospores in other fungi is discussed.

  18. Identification of Candida albicans by using different culture medias and its association in potentially malignant and malignant lesions.

    Science.gov (United States)

    Saigal, Sonal; Bhargava, Ankur; Mehra, S K; Dakwala, Falguni

    2011-07-01

    The present study evaluates the association of Candida albicans with normal control group, potentially malignant and malignant lesions of oral cavity by using two different liquid culture media. Saliva was collected and biopsy was taken only from those clinically suspected potentially malignant and malignant lesions for histopathological diagnosis. Saliva samples were inoculated for fungal growth in Sabouraud's dextrose agar and culture-positive samples had undergone for Germ tube test. Germ tube-positive samples were further taken for quantification of chlamydospore production in liquid media at 8 and 16 hours. In normal control groups no fungus growth was found; however, potentially malignant and malignant cases showed fungus growth, positive germ tube test and chlamydospore formation. The result also showed rapid and quantitatively more chlamydospore formation in corn meal broth + 5% milk in comparison to serum milk culture media. The oral mucosa is compromised in potentially malignant lesions, it can be argued that this species may be involved in carcinogenesis by elaborating the nitrosamine compounds which either act directly on oral mucosa or interact with other chemical carcinogens to activate specific proto-oncogenes and thereby initiate oral neoplasia.

  19. Identification of Candida albicans by using different culture medias and its association in potentially malignant and malignant lesions

    Directory of Open Access Journals (Sweden)

    Sonal Saigal

    2011-01-01

    Full Text Available Background and Objective: The present study evaluates the association of Candida albicans with normal control group, potentially malignant and malignant lesions of oral cavity by using two different liquid culture media. Materials and Methods: Saliva was collected and biopsy was taken only from those clinically suspected potentially malignant and malignant lesions for histopathological diagnosis. Saliva samples were inoculated for fungal growth in Sabouraud′s dextrose agar and culture-positive samples had undergone for Germ tube test. Germ tube-positive samples were further taken for quantification of chlamydospore production in liquid media at 8 and 16 hours. Results: In normal control groups no fungus growth was found; however, potentially malignant and malignant cases showed fungus growth, positive germ tube test and chlamydospore formation. The result also showed rapid and quantitatively more chlamydospore formation in corn meal broth + 5% milk in comparison to serum milk culture media. Conclusion: The oral mucosa is compromised in potentially malignant lesions, it can be argued that this species may be involved in carcinogenesis by elaborating the nitrosamine compounds which either act directly on oral mucosa or interact with other chemical carcinogens to activate specific proto-oncogenes and thereby initiate oral neoplasia.

  20. Controle biológico de helmintos parasitos de animais: estágio atual e perspectivas futuras Biological control of helminth parasites of animals: current stage and future outlook

    Directory of Open Access Journals (Sweden)

    Marcelo de Andrade Mota

    2003-09-01

    Full Text Available O controle biológico é um método desenvolvido para diminuir uma população de parasitas pela utilização de antagonista natural. A administração de fungos nematófagos aos animais domésticos é considerada uma promissora alternativa na profilaxia das helmintíases gastrintestinais parasitárias. Os fungos nematófagos desenvolvem estruturas em forma de armadilhas, responsáveis pela captura e destruição dos estágios infectantes dos nematóides. Os fungos dos gêneros Arthrobotrys, Duddingtonia e Monacrosporium têm demonstrado eficácia em experimentos laboratoriais e no campo no controle de parasitos de bovinos, eqüinos, ovinos e suínos. Diversas formulações fúngicas têm sido avaliadas, no entanto, ainda não há nenhum produto comercial disponível. A associação dos grupos de pesquisa e o envolvimento das indústrias poderão colaborar para o sucesso na implementação desta forma de controle.Biological control is a non-chemical alternative method with its main goal to reduce the amount of parasite population using natural antagonists. The administration of nematophagous fungi to animals has been considered an alternative in gastrointestinal helminthiasis prophylaxis. The nematophagous fungi produce trap-shaped structures, which are responsible for capturing and destroying the free-living stages of nematodes. The genera Arthrobotrys, Duddingtonia and Monacrosporium has been shown efficacy in laboratory and field experiments against cattle, equine, ovine and swine parasites. Several fungi formulations have been evaluated, but there is so far no commercial product available. The association of research groups with industry could improve the successful implementation of this control method.

  1. Biofilm forming cyanobacteria, algae and fungi on two historic monuments in Belgrade, Serbia

    Directory of Open Access Journals (Sweden)

    Ljaljević-Grbić Milica

    2010-01-01

    Full Text Available Biofilm on the sandstone substrata of the bridge 'Brankov most' and on the granite substrata of the 'Monument of the Unknown Hero' contains a complex consortia of cyanobacteria, algae, and fungi. Coccoid and filamentous cyanobacteria, green algae and diatoms make up the photosynthetic part of the biofilm while hyphal fragments, chlamydospores, fruiting bodies and spores take part as fungal components. These structures make a dense layer by intertwining and overlapping the stone surface. Five cyanobacterial, 11 algal and 23 fungal taxa were found. The interaction of the biofilm's constituents results in the bioweathering of the stone substrata through mechanical penetration, acid corrosion and the production of secondary mycogenic biominerals. .

  2. Scytalidium parasiticum sp. nov., a New Species Parasitizing on Ganoderma boninense Isolated from Oil Palm in Peninsular Malaysia.

    Science.gov (United States)

    Goh, Yit Kheng; Goh, Teik Khiang; Marzuki, Nurul Fadhilah; Tung, Hun Jiat; Goh, You Keng; Goh, Kah Joo

    2015-06-01

    A mycoparasite, Scytalidium parasiticum sp. nov., isolated from the basidiomata of Ganoderma boninense causing basal stem rot of oil palm in Johor, Malaysia, is described and illustrated. It is distinct from other Scytalidium species in having smaller asci and ascospores (teleomorphic stage), longer arthroconidia (anamorphic stage), hyaline to yellowish chlamydospores, and producing a fluorescent pigment. The phylogenetic position of S. parasiticum was determined by sequence analyses of the internal transcribed spacers and the small-subunit ribosomal RNA gene regions. A key to identify Scytalidium species with teleomorphic stage is provided.

  3. Differentiation between Candida albicans and Candida dubliniensis using hypertonic Sabouraud broth and tobacco agar

    Directory of Open Access Journals (Sweden)

    Fabíola Silveira-Gomes

    2011-08-01

    Full Text Available INTRODUCTION: Opportunistic fungal infections in immunocompromised hosts are caused by Candida species, and the majority of such infections are due to Candida albicans. However, the emerging pathogen Candida dubliniensis demonstrates several phenotypic characteristics in common with C. albicans, such as production of germ tubes and chlamydospores, calling attention to the development of stable resistance to fluconazole in vitro. The aim of this study was to evaluate the performance of biochemistry identification in the differentiating between C. albicans and C. dubliniensis, by phenotyping of yeast identified as C. albicans. METHODS: Seventy-nine isolates identified as C. albicans by the API system ID 32C were grown on Sabouraud dextrose agar at 30°C for 24-48h and then inoculated on hypertonic Sabouraud broth and tobacco agar. RESULTS: Our results showed that 17 (21.5% isolates were growth-inhibited on hypertonic Sabouraud broth, a phenotypic trait inconsistent with C. albicans in this medium. However, the results observed on tobacco agar showed that only 9 (11.4% of the growth-inhibited isolates produced characteristic colonies of C. dubliniensis (rough colonies, yellowish-brown with abundant fragments of hyphae and chlamydospores. CONCLUSIONS: The results suggest that this method is a simple tool for screening C. albicans and non-albicans yeast and for verification of automated identification.

  4. Differentiation between Candida albicans and Candida dubliniensis using hypertonic Sabouraud broth and tobacco agar.

    Science.gov (United States)

    Silveira-Gomes, Fabíola; Sarmento, Dayse Nogueira; Espírito-Santo, Elaine Patrícia Tavares do; Souza, Nádia de Oliveira; Pinto, Thifany Mendes; Marques-da-Silva, Silvia Helena

    2011-01-01

    Opportunistic fungal infections in immunocompromised hosts are caused by Candida species, and the majority of such infections are due to Candida albicans. However, the emerging pathogen Candida dubliniensis demonstrates several phenotypic characteristics in common with C. albicans, such as production of germ tubes and chlamydospores, calling attention to the development of stable resistance to fluconazole in vitro. The aim of this study was to evaluate the performance of biochemistry identification in the differentiating between C. albicans and C. dubliniensis, by phenotyping of yeast identified as C. albicans. Seventy-nine isolates identified as C. albicans by the API system ID 32C were grown on Sabouraud dextrose agar at 30°C for 24-48h and then inoculated on hypertonic Sabouraud broth and tobacco agar. Our results showed that 17 (21.5%) isolates were growth-inhibited on hypertonic Sabouraud broth, a phenotypic trait inconsistent with C. albicans in this medium. However, the results observed on tobacco agar showed that only 9 (11.4%) of the growth-inhibited isolates produced characteristic colonies of C. dubliniensis (rough colonies, yellowish-brown with abundant fragments of hyphae and chlamydospores). The results suggest that this method is a simple tool for screening C. albicans and non-albicans yeast and for verification of automated identification.

  5. Effect of aqueous vitamin B on the growth of blister blight pathogen, Exobasidium vexans

    Directory of Open Access Journals (Sweden)

    Hideyuki Nagao

    2012-12-01

    Full Text Available The effect of three aqueous solution of biotin, thiamine and calcium pantothenate, on the growth of Exobasidiumvexans was examined in vitro. The germination process of basidiospores of E. vexans differed from those of the otherExobasidium species. Basidiospore germination commenced after 19.5 hr incubation and chlamydospore-like bodies wereformed after 96 hr of incubation. Addition of biotin, calcium pantothenate, and thiamine to Difco PDA and Czapek’s mediumdid not affect the proportion of germinating basidiospores. The length of germ tubes was enhanced only by addition ofthiamine in the media. Larger size germ tubes (thick germ tubes were occasionally observed among the ordinary hyphae.Most germlings of basidiospores developed chlamydospore-like bodies or autolysed on the media. Thick germ tubesfrequently appeared on the calcium pantothenate amended media and developed into a colony when these hyphae weretransferred to new calcium pantothenate amended media. However, further transfer of colonies did not successfully bring anew colony to grow on the calcium pantothenate amended media. Vitamin B5, calcium pantothenate, was only partially effective in generating the thick germ tubes and to induce the initial colony formation, whereas amendment of biotin and thiamineto the media did not induce visible colony growth.

  6. Morphological identification of Candida species on glucose agar, rice extract agar and corn meal agar with and without Tween-80.

    Science.gov (United States)

    Joshi, K R; Solanki, A; Prakash, P

    1993-01-01

    A comparative study for the identification of 32 known strains of Candida species on the basis of morphology on glucose agar, rice extract agar and corn meal agar with and without Tween 80 revealed that when Tween 80 is incorporated in the media identification is possible for 96.8% of the species within 48 hours on rice extract agar and for 96.8% of the species within 48 hours on rice extract agar and for 90.6% of the species on glucose agar. The germ tubes and chlamydospores were also produced more on rice extract agar than on 0.1% glucose agar. Rice extract agar with Tween 80 can be used as single medium for morphologic identification of Candida species. The inoculated medium is first incubated at 37 degrees C for 3 hours and examined for germ tube formation and then incubated at 25 degrees C for 24 to 72 hours and examined for appearance of chlamydospores and mycelial morphology.

  7. High prevalence of oral colonization by Candida dubliniensis in HIV-positive patients in Argentina.

    Science.gov (United States)

    Binolfi, Andrés; Biasoli, Marisa S; Luque, Alicia G; Tosello, María E; Magaró, Hortensia M

    2005-08-01

    Candida dubliniensis is a recently described yeast species, closely related to Candida albicans. This work represents the first general survey of the carriage of C. dubliniensis in the oral cavities of HIV-positive patients in Argentina. We studied 133 strains isolated from 162 HIV-positive patients, using the following identification tests: chlamydospore production on corn meal agar with Tween 80; colony color on CHROMagar Candida media; differential growth at 45 degrees C on potato dextrose agar; D-xylose assimilation; chlamydospore formation on sunflower seed agar (SSA); carbohydrate assimilation profiles using the API 20 C Aux commercial kit and PCR using primers that hybridize to the class IV intron of the ACT1 gene. Out of the 133 strains, 21 were identified as C. dubliniensis, representing approximately 13% of the 162 patients in this study. From these data, we conclude that although the PCR assay is the most reliable method, clamydospore formation on SSA is an easier and less expensive test for the screening of C. dubliniensis in the routine laboratory. Our results show that C. dubliniensis has a high prevalence among HIV-positive patients in Argentina.

  8. Effect of different concentrations of dl-isoleucine, dl-valine, and dl-alanine on growth and sporulation in Fusarium oxysporum f. udum (Butl.) Sn. et H.

    Science.gov (United States)

    Prasad, M; Chaudhary, S K

    1977-01-01

    D1-alanine and dl-valine, when added as an extra nitrogen for fortifying the already present inorganic nitrogen source, actually acted as growth retardant for F. oxysporum f. udum (Butl.) Sn. et H. Sporulation of microconidia was indifferently affected by these two amino acids. DI-valine stimulated microconidial formation in young cultures only. In both young and old cultures the lowest concentration of dl-valine depressed macronidial sporulation. In old cultures the lowest concentration of valine stimulated chlamydospore differentiation rapidly, higher concentrations being less effective. D1-alanine, as an additional nitrogen source, depressed both macro- and microconidal sporulation. It did not even invigorate chlamydospore formation. D1-isoleucine, on the other hand, belongs to the category of growth promoters and profuse and stimulative sporulators of macro- and microconidia. This pathogen needs very specific and preferential doses of the three amino acids, if these are used as a booster in addition to the already present nitrogen source. The response, both in terms of mycelial growth and sporulation of the three spore forms, was also conditioned by the age of the culture.

  9. Pochonia chlamydosporia promotes the growth of tomato and lettuce plants

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    Rosangela Dallemole-Giaretta

    2015-10-01

    Full Text Available The fungus Pochonia chlamydosporia is one of the most studied biological agents used to control plant-parasitic nematodes. This study found that the isolates Pc-3, Pc-10 and Pc-19 of this fungus promote the growth of tomato and lettuce seedlings. The isolate Pc-19 colonized the rhizoplane of tomato seedlings in only 15 days and produced a large quantity of chlamydospores. This isolate was able to use cellulose as a carbon source, in addition to glucose and sucrose. Scanning electron microscopy (SEM revealed that hyphae of the P. chlamydosporia isolate Pc-10 penetrated the epidermal cells of the tomato roots. These three P. chlamydosporia isolates promote the growth of tomato and lettuce.

  10. Hichrom candida agar for identification of Candida species.

    Science.gov (United States)

    Baradkar, V P; Mathur, M; Kumar, S

    2010-01-01

    Chromogenic media are frequently used in direct and rapid identification of yeasts because different Candida species produce unique colors on these media. We used 60 isolates of Candida species including 30 C. albicans, 10 C. parapsilosis, 11 C. glabrata, five C. tropicalis, and four C. dubliniensis, isolated from various clinical specimens, to evaluate the performance of HiChrome Candida agar. These strains had been identified by germ tube test, morphology on cornmeal agar, chlamydospore formation on tobacco agar and sugar assimilation tests. The sensitivity and specificity results were: C. albicans (96.55 and 96.42%); C. parapsilosis (80 and 98.03%), C. glabrata (90.90 and 88.23%), C. tropicalis (100 and 100%) and C. dubliniensis (60 and 96.55%) respectively. HiChrom Candida agaris medium has been useful and capable of presumptive, rapid identification of Candida species within 48 hours.

  11. Hichrom candida agar for identification of candida species

    Directory of Open Access Journals (Sweden)

    Baradkar V

    2010-01-01

    Full Text Available Chromogenic media are frequently used in direct and rapid identification of yeasts because different Candida species produce unique colors on these media. We used 60 isolates of Candida species including 30 C. albicans, 10 C. parapsilosis, 11 C. glabrata, five C. tropicalis, and four C. dubliniensis, isolated from various clinical specimens, to evaluate the performance of HiChrome Candida agar. These strains had been identified by germ tube test, morphology on cornmeal agar, chlamydospore formation on tobacco agar and sugar assimilation tests. The sensitivity and specificity results were: C. albicans (96.55 and 96.42%; C. parapsilosis (80 and 98.03%, C. glabrata (90.90 and 88.23%, C. tropicalis (100 and 100% and C. dubliniensis (60 and 96.55% respectively. HiChrom Candida agaris medium has been useful and capable of presumptive, rapid identification of Candida species within 48 hours.

  12. [A case of Tinea capitis caused by Trichophyton tonsurans].

    Science.gov (United States)

    Urano, Shoko; Shirai, Shigeko; Suzuki, Yoko; Sugaya, Keiko; Takigawa, Masahiro; Mochizuki, Takashi

    2003-01-01

    A 10-year-old Peruvian girl, living in Japan since 1996, visited our hospital in August 2000 complaining of alopecia which had been present on her scalp for one year. The bald areas appeared as multiple small, scattered, angular patches with indistinct margins. Follicular pustules, erythemic nodules and lymphadenopathy were also seen. In the culture of the affected hair, a tan surface with wiry undulations grew on Sabouraud's media. The colony reverse had reddish-brown central pigmentation. Slide cultured fungi produced great numbers of round and short club-shaped microconidia, hyphae and intercalary chlamydospores. These fungi showed the following characteristics: positive urease test, no pigment production on cornmeal agar and positive thiamine dependency. The restriction fragment length polymorphism pattern and the nucleotide sequences of ribosomal-DNA internal transcribed spacer region of the causative fungus was compatible with Trichophyton tonsurans. Daily administration of 125 mg of terbinafine resulted in a satisfactory response and the lesion healed almost completely.

  13. Murine model for Fusarium oxysporum invasive fusariosis reveals organ-specific structures for dissemination and long-term persistence.

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    Katja Schäfer

    Full Text Available The soil-borne plant pathogen Fusarium oxysporum causes life-threatening invasive fusariosis in immunocompromised individuals. The mechanism of infection in mammalian hosts is largely unknown. In the present study we show that the symptoms of disseminated fusariosis caused by F. oxysporum in immunosuppressed mice are remarkably similar to those reported in humans. Distinct fungal structures were observed inside the host, depending on the infected organ. Invasive hyphae developed in the heart and kidney, causing massive colonization of the organs. By contrast, chlamydospore-like survival structures were found in lung, spleen and liver. Systemically infected mice also developed skin and eye infections, as well as thrombosis and necrosis in the tail. We further show that F. oxysporum can disseminate and persist in the organs of immunocompetent animals, and that these latent infections can lead to lethal systemic fusariosis if the host is later subjected to immunosuppressive treatment.

  14. Three New Soil-inhabiting Species of Trichoderma in the Stromaticum Clade with Test of Their Antagonism to Pathogens.

    Science.gov (United States)

    Chen, Kai; Zhuang, Wen-Ying

    2017-09-01

    Trichoderma is a dominant component of the soil mycoflora. During the field investigations of northern, central, and southwestern China, three new species in the Stromaticum clade were encountered from soil, and named as T. hebeiense, T. sichuanense, and T. verticillatum. Their phylogenetic positions were determined by analyses of the combined two genes: partial sequences of translation elongation factor 1-alpha and the second largest RNA polymerase subunit-encoding genes. Distinctions between the new species and their close relatives were discussed. Trichoderma hebeiense appeared as a separate terminal branch. The species is distinctive by its oblong conidia and aggregated pustules in culture. Trichoderma sichuanense features in concentric colony and produces numerous clean exudates on aerial mycelium in culture. Trichoderma verticillatum is characterized by its verticillium-like synanamorph and production of abundant chlamydospores. In vitro antagonism towards the new species was tested by dual culture technique.

  15. Isolation of Candida dubliniensis for the first time in Cali, Colombia, and its identification with phenotyping methods.

    Science.gov (United States)

    Alvarez, María Inés; Suárez, Blanca Lynne; Caicedo, Luz Dary

    2009-01-01

    Candida dubliniensis is an emerging pathogenic yeast isolated mainly from the oral cavity of HIV-infected patients. The close phenotypic and genotypic relationship between C. albicans and C. dubliniensis has led to incorrectly identifying isolates of C. dubliniensis as C. albicans. The oral cavities of 107 diabetic patients were studied in Cali, Colombia, and 72 colonies of Candida, with shades of green on CHROMagar Candida culture media, were obtained. Various phenotypic tests were carried out, which included germ tube formation and production of chlamydospores on corn meal Agar. Additionally, growth studies were carried out at 42 degrees C and 45 degrees C and on Sabouraud agar with 6.5%, sodium chloride. Identification of C. dubliniensis with these tests was confirmed with API 20C Aux. We identified 65 and 7 colonies of C. albicans and C. dubliniensis, respectively. This is the first time that C. dubliniensis is identified with phenotypic methods in Colombia.

  16. Characterization of Italian isolates of Inonotus rickii

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    Tiziana ANNESI

    2011-01-01

    Full Text Available Thirty-seven isolates of Inonotus rickii, a pathogenic fungus causing white rot and cankers, were collected from diseased boxelder trees lining boulevards in Rome and from other hosts in Rome and Sicily. During the survey, it was observed that this fungus occasionally produced basidiomes, but more frequently it had anamorphic structures that released a brown powdery mass of chlamydospores, presumably acting as asexual propagules. All isolates were characterized using random amplified microsatellite analysis and somatic incompatibility tests in order to investigate the diversity of genotypes within and between the different disease centers surveyed in Italy. The results suggest that both sexual and asexual reproduction play an important role in the spread of this disease, with important epidemiological implications. .

  17. Comparative studies with regard to the influence of carbon and nitrogen ratio on sporulation in Fusarium oxysporum and Fusarium moniliforme v. subglutinans.

    Science.gov (United States)

    Prasad, M

    1979-01-01

    Carbon/nitrogen ratio as a factor for sporulation, expressed in terms of magnitude of population variation of macroconidia and microconidia in the cultures of Eusarium oxysporum Schlecht ex. Fr., Fusarium moniliforme v. subglutinans Wr. and Rg., and of chlamydospores (only in Fusarium oxysporum) was investigated. It has been found that the amount of carbon source shapes the course of macro- and micro. conidial production in a linear fashion, being enhanced parallel to the increase in its amount-Nitrogen level, limiting proliferation and effectively diminishing the macro- and micro-conidial population, varies for the two species, namely Fusarium oxysporum and Fusarium moniliforme v-subglutinans. For chlamydomspore production, higher carbon and still higher nitrogen concentration favours profuse proliferation in case of Fusarium oxysporum.

  18. [Identification of Candida dubliniensis strains using heat tolerance tests, morphological characteristics and molecular methods].

    Science.gov (United States)

    Arikan, Sevtap; Darka, Ozge; Hasçelik, Gülşen; Günalp, Ayfer

    2003-01-01

    Described in 1995, Candida dubliniensis is a novel Candida species closely related to Candida albicans due primarily to its ability to produce germ tube and chlamydospores. Given these phenotypic similarities between the two species, C. dubliniensis cannot be readily distinguished from Candida albicans by routine laboratory work-up. We explored the frequency of isolation of C. dubliniensis among 213 strains previously defined as C. albicans based on their ability to produce germ tube. The test isolates were initially examined for their morphological features on cornmeal tween 80 agar, inability to grow at 45 degrees C, and the biochemical assimilation profile (ID 32C system, bioMerieux, France). Among all, 2 (0.9%) of the isolates were identified as C. dubliniensis based on the production of numerous chlamydospores in chains on cornmeal tween 80 agar and the lack of growth at 45 degrees C. The assimilation profile of these isolates was found to be in accordance with this identification. In an effort to confirm the identification, polymerase chain reaction (PCR) studies were carried out by using the C. dubliniensis specific primer set, DUBF and DUBR. Both of the isolates yielded C. dubliniensis-specific 288 base pair amplification products, confirming the previous identification obtained with the initial screening tests. The isolates were found to be susceptible to fluconazole and itraconazole, and generated amphotericin B minimal inhibitory concentrations of 0.5-1 microgram/ml by NCCLS M27-A2 microdilution method. These data suggest that the isolation rate of C. dubliniensis among our clinical isolates is low. The morphological features on cornmeal tween 80 agar and the lack of ability to grow at 45 degrees C appear as reliable, cheap, and practical screening tests in initial identification of C. dubliniensis among germ tube-producing Candida strains.

  19. Biocontrol potential of Pasteuria penetrans, Pochonia chlamydosporia, Paecilomyces lilacinus and Trichoderma harzianum against Meloidogyne incognita in

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    Tariq MUKHTAR

    2013-05-01

    Full Text Available The root-knot nematode, Meloidogyne incognita, is a sedentary endoparasitic plant pathogen with a very wide host range, which causes annual crop losses amounting to millions of dollars. The small number of available nematicides and restrictions on the use of non-fumigant nematicides due to high toxicity to humans and non-target organisms hinder effective nematode control. A possible alternative to chemical nematicides is the use of biological control agents for the management of this nematode. In the present study, the efficacy of four biocontrol agents was tested against M. incognita at different doses. The biocontrol agents Pasteuria penetrans, Pochonia chlamydosporia, Paecilomyces lilacinus and Trichoderma harzianum were mass produced and mixed with the formalin sterilized soil at the rates of 2 × 103, 4 × 103, 6 × 103, 8 × 103, and 1 × 104 endospores/chlamydospores/cfu per g of soil. Okra seeds (cv. Sabz Pari were sown in pots of soil amended with the different agents, and 10 d after emergence, the plants were inoculated with 2000 freshly hatched second stage juveniles of M. incognita. Data on plant growth parameters and nematode infestations were recorded 7 weeks after inoculation. The antagonists varied significantly in enhancing various growth parameters and reducing nematode infestations in a dose-responsive manner. Both P. penetrans and P. lilacinus were equally effective and caused maximum reductions in number of galls, egg masses, nematode fecundity and build up as compared with T. harzianum and P. chlamydosporia. Reductions in these parameters at the concentration of 8 × 103 were statistically similar with those caused at the concentration of 1 × 104 chlamydospores/ endospores/cfu. Our results indicate that application of antagonists can suppress galling and reproduction of M. incognita resulting in enhancement of plant growth.

  20. The effect of some organic substances on the mycelium of the fungus Ustilago nuda (Jens.) Rostr.

    Science.gov (United States)

    Krátká, J

    1976-01-01

    Research was performed for studying the effect of some organic compounds, considered by many authors as the products ob barley seed metabolism generated after anaerobic seed treatment, on the mycelium of the fungus Ustilago nuda (Jens.) Rostr. The author examined the effectiveness of ethylacohol, acetaldehyde, acetic acid, succinic acid, lactic acid, and hydroquinone in concentrations from 1 M to 10(-6) M, and the effectiveness of extracts from disinfected seeds in doses from 10 g to 0.001 g/l. The effect of the mentioned solutions was examined as exerted on the growth of dicaryotic mycelium and on the growth of the haploid promycelium of the fungus. The dicaryotic mycelium of Ustilago nuda (Jens.) Rostr. was cultivated on potato agar with benzoic acid. The presence of the acid prevents mitosis, and the chlamydospores germinate on the nutritive medium with two fibres having binuclear cells. The haploid promycelium was cultivated on potato agar; chlamydospores germinated with one four-cell fibre, and individual cells are mononuclear and haploid. Only later, a dicarytic mycelium is created in a complex process. In all the substances used, the concentration of 1 M was found to stop further growth of mycelium. The concentration of 10(-1) M of acetic acid and hydroquinone also stopped growth, the same concentration of acetaldehyde, lactic acid, succinic acid, ethylacohol stimulated mycelium growth in comparison with the control. The concentration of 10(-6) M stimulated mycelium growth in a majority of cases. Extracts from disinfected seeds did not influence mycelium growth significantly in all cases in comparison with the control. The results were similar in the two types of mycelium.

  1. Mutualism in a Reduced Gravity Environment (MuRGE)

    Science.gov (United States)

    Haire, Timothy C.

    2010-01-01

    Mutualism in a Reduced Gravity Environment (MuRGE) is a ground research study to determine the feasibility of assessing fungi-plant (Piriformospora indica-Arabidopsis thaliana) interactions in microgravity. Seeds from the plant Arabiddospsis thaliana (At) will be grown in the presence of Piriformospora indica (Pi) an endophytic Sebacinacae family fungus. Pi is capable of colonizing the roots of a wide variety of plant species, including non-mycorrhizal hosts like At, and promoting plant growth similarly to AMF (arbusuclar mychorrizal fungi) unlike most AMF, Pi is not an obligate plant symbiont and can be grown in the absence of a host. In the presence of a suitable plant host, Pi can attach to and colonize root tips. Interaction visualization is accomplished with strong autofluorescence in the roots, followed by root colonization via fungal hyphae, and chlamydospore production. Increased root growth can be observed even before root colonization is detectable. In addition, Pi chlamydospores generated from axenic culture in microgravity will be used to inoculate roots of At grown in 1g to determine the effect of microgravity upon the inherent virulence or beneficial effects. Based on recent reports of increased virulence of S. typhimurium, P. aeruginosa, and S. Pneumoniae in reduced gravity, differences in microbial pathogenic responses and host plant systemic acquired resistance are expected. The focus of this project within MuRGE involved the development P. indica culture media evaluation and microscopy protocol development. High, clean spore harvest yields for the detection of fungi-plant interactions microscopically was the immediate goal of this experiment.

  2. Conserved Responses in a War of Small Molecules between a Plant-Pathogenic Bacterium and Fungi.

    Science.gov (United States)

    Spraker, Joseph E; Wiemann, Philipp; Baccile, Joshua A; Venkatesh, Nandhitha; Schumacher, Julia; Schroeder, Frank C; Sanchez, Laura M; Keller, Nancy P

    2018-05-22

    Small-molecule signaling is one major mode of communication within the polymicrobial consortium of soil and rhizosphere. While microbial secondary metabolite (SM) production and responses of individual species have been studied extensively, little is known about potentially conserved roles of SM signals in multilayered symbiotic or antagonistic relationships. Here, we characterize the SM-mediated interaction between the plant-pathogenic bacterium Ralstonia solanacearum and the two plant-pathogenic fungi Fusarium fujikuroi and Botrytis cinerea We show that cellular differentiation and SM biosynthesis in F. fujikuroi are induced by the bacterially produced lipopeptide ralsolamycin (synonym ralstonin A). In particular, fungal bikaverin production is induced and preferentially accumulates in fungal survival spores (chlamydospores) only when exposed to supernatants of ralsolamycin-producing strains of R. solanacearum Although inactivation of bikaverin biosynthesis moderately increases chlamydospore invasion by R. solanacearum , we show that other metabolites such as beauvericin are also induced by ralsolamycin and contribute to suppression of R. solanacearum growth in vitro Based on our findings that bikaverin antagonizes R. solanacearum and that ralsolamycin induces bikaverin biosynthesis in F. fujikuroi , we asked whether other bikaverin-producing fungi show similar responses to ralsolamycin. Examining a strain of B. cinerea that horizontally acquired the bikaverin gene cluster from Fusarium , we found that ralsolamycin induced bikaverin biosynthesis in this fungus. Our results suggest that conservation of microbial SM responses across distantly related fungi may arise from horizontal transfer of protective gene clusters that are activated by conserved regulatory cues, e.g., a bacterial lipopeptide, providing consistent fitness advantages in dynamic polymicrobial networks. IMPORTANCE Bacteria and fungi are ubiquitous neighbors in many environments, including

  3. The rhizosphere microbial community in a multiple parallel mineralization system suppresses the pathogenic fungus Fusarium oxysporum

    Science.gov (United States)

    Fujiwara, Kazuki; Iida, Yuichiro; Iwai, Takashi; Aoyama, Chihiro; Inukai, Ryuya; Ando, Akinori; Ogawa, Jun; Ohnishi, Jun; Terami, Fumihiro; Takano, Masao; Shinohara, Makoto

    2013-01-01

    The rhizosphere microbial community in a hydroponics system with multiple parallel mineralization (MPM) can potentially suppress root-borne diseases. This study focused on revealing the biological nature of the suppression against Fusarium wilt disease, which is caused by the fungus Fusarium oxysporum, and describing the factors that may influence the fungal pathogen in the MPM system. We demonstrated that the rhizosphere microbiota that developed in the MPM system could suppress Fusarium wilt disease under in vitro and greenhouse conditions. The microbiological characteristics of the MPM system were able to control the population dynamics of F. oxysporum, but did not eradicate the fungal pathogen. The roles of the microbiological agents underlying the disease suppression and the magnitude of the disease suppression in the MPM system appear to depend on the microbial density. F. oxysporum that survived in the MPM system formed chlamydospores when exposed to the rhizosphere microbiota. These results suggest that the microbiota suppresses proliferation of F. oxysporum by controlling the pathogen's morphogenesis and by developing an ecosystem that permits coexistence with F. oxysporum. PMID:24311557

  4. Phytophthora pseudopolonica sp. nov., a new species recovered from stream water in subtropical forests of China.

    Science.gov (United States)

    Li, Wen-Wen; Zhao, Wen-Xia; Huai, Wen-Xia

    2017-09-01

    A new species of the genus Phytophthora was isolated from stream water in the subtropical forests of China during a survey of forest Phytophthora from 2011 to 2013. This new species is formally described here and named Phytophthora pseudopolonica sp. nov. This new homothallic species is distinct from other known Phytophthora species in morphology and produces nonpapillate and noncaducous sporangia with internal proliferation. Spherical hyphal swellings and thin-walled chlamydospores are abundant when the species is kept in sterile water. The P. pseudopolonica sp. nov. forms smooth oogonia with paragynous and sometimes amphigynous antheridia. The optimum growth temperature of the species is 30 °C in V8-juice agar with β-sitosterol, yet it barely grows at 5 °C and 35 °C. Based on sequences of the internal transcribed spacer and the combined β-tubulin and elongation factor 1α gene sequence data, isolates of the new species cluster together into a single branch and are close to Phytophthora polonicabelonging to clade 9.

  5. A high-temperature tolerant species in clade 9 of the genus Phytophthora: P. hydrogena sp. nov.

    Science.gov (United States)

    Yang, Xiao; Gallegly, Mannon E; Hong, Chuanxue

    2014-01-01

    A previously unknown Phytophthora species was isolated from irrigation water in Virginia, USA. This novel species produces abundant noncaducous and nonpapillate sporangia in soil water extract solution. It sometimes produces chlamydospores and hyphal swellings in aged cultures and in Petri's solution. This species has optimum vegetative growth at 30 C and grows well at 35 C. The lowest and highest temperatures for growth are 5 and 40 C. All isolates examined in this study are compatibility type A1 and produce mostly plerotic oospores when paired with an A2 mating-type tester of P. cinnamomi. Sequence analyses of the rDNA internal transcribed spacer (ITS) regions and the mitochondrially encoded cytochrome c oxidase 1 (cox 1) gene placed this species in clade 9 of the genus Phytophthora. These characteristics support the description of this taxon as a new species for which we propose the name P. hydrogena sp. nov. Further phylogenetic and physiological investigations of clade 9 species revealed a high-temperature tolerant cluster including P. hydrogena, P. aquimorbida, P. hydropathica, P. irrigata, P. chrysanthemi, P. insolita, P. polonica and P. parsiana. These species all grow well at 35 C. The monophyly of the species in this heat-tolerant cluster except P. insolita and P. polonica is highly supported by the maximum-likelihood analyses of the ITS and cox 1 sequences.

  6. A unique species in Phytophthora clade 10, Phytophthora intercalaris sp. nov., recovered from stream and irrigation water in the eastern USA

    Science.gov (United States)

    Balci, Y.; Brazee, N. J.; Loyd, A. L.; Hong, C. X.

    2016-01-01

    A novel species of the genus Phytophthora was recovered during surveys of stream and nursery irrigation water in Maryland, Massachusetts, North Carolina, Virginia and West Virginia in the USA. The novel species is heterothallic, and all examined isolates were A1 mating type. It produced rare ornamented oogonia and amphigynous antheridia when paired with A2 mating type testers of Phytophthora cinnamomi and Phytophthora cryptogea. Sporangia of this novel species were non-papillate and non-caducous. Thin-walled intercalary chlamydospores were abundant in hemp seed agar and carrot agar, while they were produced only rarely in aged cultures grown in clarified V8 juice agar. Phylogenetic analyses based on sequences of the internal transcribed spacer region and the β-tubulin and mitochondrial cytochrome-c oxidase 1 (cox1) genes indicated that the novel species is phylogenetically close to Phytophthora gallica in Phytophthora clade 10. The novel species has morphological and molecular features that are distinct from those of other species in Phytophthora clade 10. It is formally described here as Phytophthora intercalaris sp. nov. Description of this unique clade-10 species is important for understanding the phylogeny and evolution of Phytophthora clade 10. PMID:26620125

  7. Phytophthora ×stagnum nothosp. nov., a New Hybrid from Irrigation Reservoirs at Ornamental Plant Nurseries in Virginia

    Science.gov (United States)

    Yang, Xiao; Richardson, Patricia A.; Hong, Chuanxue

    2014-01-01

    A novel Phytophthora species was frequently recovered from irrigation reservoirs at several ornamental plant production facilities in eastern Virginia. Initial sequencing of the internal transcribed spacer (ITS) region of this species generated unreadable sequences due to continual polymorphic positions. Cloning and sequencing the ITS region as well as sequencing the mitochondrially encoded cytochrome c oxidase 1 and beta-tubulin genes revealed that it is a hybrid between P. taxon PgChlamydo as its paternal parent and an unknown species genetically close to P. mississippiae as its maternal parent. This hybrid has some diagnostic morphological features of P. taxon PgChlamydo and P. mississippiae. It produces catenulate hyphal swellings, characteristic of P. mississippiae, and chlamydospores, typical of P. taxon PgChlamydo. It also produces both ornamented and relatively smooth-walled oogonia. Ornamented oogonia are another important diagnostic character of P. mississippiae. The relatively smooth-walled oogonia may be indicative of oogonial character of P. taxon PgChlamydo. The new hybrid is described here as Phytophthora ×stagnum. PMID:25072374

  8. Evaluation of SOC for the presumptive identification of Candida albicans and Cryptococcus neoformans.

    Science.gov (United States)

    Fleming, W H; Knezek, K L; Dorn, G L

    1987-01-01

    SOC, a fungal growth medium composed of Solryth, oxgall, and caffeic acid, was evaluated as a medium to provide rapid, differential identification of Candida albicans and Cryptococcus neoformans. Using a variety of common isolation media to produce the yeast inocula, the germ tube methods tested ranked in the following order of decreasing sensitivity: SOC (97% +/- 1), serum (92% +/- 5), rabbit coagulase plasma with EDTA in combination with tryptic soy broth (89% +/- 5), TOC (89% +/- 6), and rabbit coagulase plasma with EDTA (83% +/- 4). In chlamydospore production, SOC also proved to be the most sensitive after 24 h incubation: SOC (96% +/- 2), TOC (80% +/- 2), and cornmeal-Tween 80 agar (14% +/- 3). Other medically important yeasts showed normal patterns of growth within 24 h on SOC, thus assisting in their identification. Eighty strains of Cryptococcus neoformans showed characteristic brown pigmentation on SOC and TOC within 18 h, while all other species of the genus Cryptococcus and 229 Candida isolates did not show a change in pigmentation.

  9. Predatory Capacity in vitro of Native Nematophagous Fungi from Cundinamarca on Gastrointestinal Nematodes of Cattle

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    Dildo Márquez Lara

    2015-12-01

    Full Text Available Dependence and indiscriminate use of chemical anthelmintics as the sole method for controlling gastrointestinal nematodes (GIN of cattle causes problems in the environment, public health, and the productivity of cattle. It is important to develop non-chemical control strategies. Nematophagous fungi can be a viable and promising alternative for the control of these endoparasites. This study aimed to isolate, identify and evaluate in vitro the potential of nematophagous fungi from Cundinamarca on L3 larvae of gastrointestinal nematodes of cattle. 60 soil samples from cattle ranches were sown in Petri boxes containing agar-water for trapping fungi, and three strains of the fungus Arthrobotrys oligospora (L1, XVIII, and XXI and one of Arthrobotrys musiformis (XXIV were identified by morphometric keys. 1 x 106 conidia or chlamydospores of each fungi were used, which faced 100 nematode larvae. Isolate XXIV (A. musiformis showed greater predatory capacity (96.8% than isolates (A. oligospora XVIII, L1, and XXI (69.68, 71.1, and 87.62%, respectively. There were no statistically significant differences (p > 0.05 among the strains with more predatory capacity. This is the first record of in vitro identification and evaluation of the predatory capacity of A. oligospora and A. musiformis, native fungi from Cundinamarca. The results suggest that these fungi could be used as biocontrol agents of nematodes in cattle.

  10. Two new hyaline-ascospored species of Trichoderma and their phylogenetic positions.

    Science.gov (United States)

    Qin, W T; Zhuang, W Y

    2016-01-01

    Collections of hypocrealean fungi found on decaying wood in subtropical regions of China were examined. Two new species, Trichoderma confluens and T. hubeiense, were discovered and are described. Trichoderma confluens is characterized by its widely effuse to rarely pulvinate, yellow stromata with densely disposed yellowish brown ostioles, simple acremonium- to verticillium-like conidiophores, hyaline conidia and multiform chlamydospores. Trichoderma hubeiense has pulvinate, grayish yellow stromata with brownish ostioles, trichoderma- to verticillium-like conidiophores and hyaline conidia. The phylogenetic positions of the two fungi were investigated based on sequence analyses of RNA polymerase II subunit b and translation elongation factor 1-α genes. The results indicate that T. confluens belongs to the Hypocreanum clade and is associated with but clearly separated from T. applanatum and T. decipiens. Trichoderma hubeiense belongs to the Polysporum clade and related to T. bavaricum but obviously differs from other members of the clade in sequence data. Morphological distinctions between the new species and their close relatives are noted and discussed. © 2016 by The Mycological Society of America.

  11. Regulation of morphogenesis and biocontrol properties in Trichoderma virens by a VELVET protein, Vel1.

    Science.gov (United States)

    Mukherjee, Prasun K; Kenerley, Charles M

    2010-04-01

    Mycoparasitic strains of Trichoderma are applied as commercial biofungicides for control of soilborne plant pathogens. Although the majority of commercial biofungicides are Trichoderma based, chemical pesticides, which are ecological and environmental hazards, still dominate the market. This is because biofungicides are not as effective or consistent as chemical fungicides. Efforts to improve these products have been limited by a lack of understanding of the genetic regulation of biocontrol activities. In this study, using gene knockout and complementation, we identified the VELVET protein Vel1 as a key regulator of biocontrol, as well as morphogenetic traits, in Trichoderma virens, a commercial biocontrol agent. Mutants with mutations in vel1 were defective in secondary metabolism (antibiosis), mycoparasitism, and biocontrol efficacy. In nutrient-rich media they also lacked two types of spores important for survival and development of formulation products: conidia (on agar) and chlamydospores (in liquid shake cultures). These findings provide an opportunity for genetic enhancement of biocontrol and industrial strains of Trichoderma, since Vel1 is very highly conserved across three Trichoderma species.

  12. Regulation of Morphogenesis and Biocontrol Properties in Trichoderma virens by a VELVET Protein, Vel1▿ †

    Science.gov (United States)

    Mukherjee, Prasun K.; Kenerley, Charles M.

    2010-01-01

    Mycoparasitic strains of Trichoderma are applied as commercial biofungicides for control of soilborne plant pathogens. Although the majority of commercial biofungicides are Trichoderma based, chemical pesticides, which are ecological and environmental hazards, still dominate the market. This is because biofungicides are not as effective or consistent as chemical fungicides. Efforts to improve these products have been limited by a lack of understanding of the genetic regulation of biocontrol activities. In this study, using gene knockout and complementation, we identified the VELVET protein Vel1 as a key regulator of biocontrol, as well as morphogenetic traits, in Trichoderma virens, a commercial biocontrol agent. Mutants with mutations in vel1 were defective in secondary metabolism (antibiosis), mycoparasitism, and biocontrol efficacy. In nutrient-rich media they also lacked two types of spores important for survival and development of formulation products: conidia (on agar) and chlamydospores (in liquid shake cultures). These findings provide an opportunity for genetic enhancement of biocontrol and industrial strains of Trichoderma, since Vel1 is very highly conserved across three Trichoderma species. PMID:20154111

  13. The rhizosphere microbial community in a multiple parallel mineralization system suppresses the pathogenic fungus Fusarium oxysporum.

    Science.gov (United States)

    Fujiwara, Kazuki; Iida, Yuichiro; Iwai, Takashi; Aoyama, Chihiro; Inukai, Ryuya; Ando, Akinori; Ogawa, Jun; Ohnishi, Jun; Terami, Fumihiro; Takano, Masao; Shinohara, Makoto

    2013-12-01

    The rhizosphere microbial community in a hydroponics system with multiple parallel mineralization (MPM) can potentially suppress root-borne diseases. This study focused on revealing the biological nature of the suppression against Fusarium wilt disease, which is caused by the fungus Fusarium oxysporum, and describing the factors that may influence the fungal pathogen in the MPM system. We demonstrated that the rhizosphere microbiota that developed in the MPM system could suppress Fusarium wilt disease under in vitro and greenhouse conditions. The microbiological characteristics of the MPM system were able to control the population dynamics of F. oxysporum, but did not eradicate the fungal pathogen. The roles of the microbiological agents underlying the disease suppression and the magnitude of the disease suppression in the MPM system appear to depend on the microbial density. F. oxysporum that survived in the MPM system formed chlamydospores when exposed to the rhizosphere microbiota. These results suggest that the microbiota suppresses proliferation of F. oxysporum by controlling the pathogen's morphogenesis and by developing an ecosystem that permits coexistence with F. oxysporum. © 2013 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  14. First report of Fusarium oxysporum species complex infection in zebrafish culturing system.

    Science.gov (United States)

    Kulatunga, D C M; Dananjaya, S H S; Park, B K; Kim, C-H; Lee, J; De Zoysa, M

    2017-04-01

    Fusarium oxysporum species complex (FOSC) is a highly diverse fungus. Recently, F. oxysporum infection was identified from zebrafish (Danio rerio) culturing system in Korea. Initially, a rapid whitish smudge was appeared in the water with the fungal blooming on walls of fish tanks. Microscopic studies were conducted on fungal hyphae, colony pigmentation and chlamydospore formation and the presence of macro- and microspores confirmed that the isolated fungus as F. oxysporum. Furthermore, isolated F. oxysporum was confirmed by internal transcribed spacer sequencing which matched (100%) to nine F. oxysporum sequences available in GenBank. Experimental hypodermic injection of F. oxysporum into adult zebrafish showed the development of fungal mycelium and pathogenicity similar to signs observed. Histopathologic results revealed a presence of F. oxysporum hyphae in zebrafish muscle. Fusarium oxysporum growth was increased with sea salt in a concentration-dependent manner. Antifungal susceptibility results revealed that F. oxysporum is resistant to copper sulphate (up to 200 μg mL -1 ) and sensitive to nystatin (up to 40 μg mL -1 ). This is the first report of FOSC from zebrafish culture system, suggesting it appears as an emerging pathogen, thus posing a significant risk on zebrafish facilities in the world. © 2016 John Wiley & Sons Ltd.

  15. In vitro antifungal sensitivity of fluconazole, clotrimazole and nystatin against vaginal candidiasis in females of childbearing age.

    Science.gov (United States)

    Khan, Fouzia; Baqai, Rakhshanda

    2010-01-01

    Vaginal candidiasis is the most common infection of females. A large variety of antifungal drugs are used for treatment. The objective of this study was isolation and identification of Candida from high vaginal swabs and in vitro antifungal activity of Clotrimazole, Fluconazole and Nystatin against Candida. Two hundred and fifty high vaginal swabs were collected from females reporting at different hospitals of Karachi. Wet mount was performed to observe the budding cells of Candida. Vaginal swabs were cultured on Sabouraud's dextrose agar with added antibiotics. Plates were incubated at room temperature for seven days. Chlamydospores of Candida albicans were identified on corn meal agar. Species of Candida were identified on Biggy agar. In vitro antifungal activity of Clotrimazole, Fluconazole and Nystatin was performed by MIC (Minimum inhibitory concentration), well diffusion method and disc diffusion method. Out of 250 high vaginal swabs, Candida species were isolated in 100 (40%) of cases. Out of 100, C. albican 30 (30%), C. tropicalis 21 (21%), C. parapsillosis 10 (10%), C. parakrusi 8 (8%), C. glabrata 8 (8%), C. krusei 3 (3%) were isolated. In vitro antifungal activity indicated Clotrimazole (MIC 16 and 8 microg/ml) effective against 68 (70%) of Candida SPP, Fluconazole (MIC 64 and 32 microg/ml) effective against 29 (36.2%) and Nystatin disc (100 units) was 51 (63.5%) effective. C. albicans was mainly isolated. Clotrimazole was more effective as compared to Fluconazole and Nystatin. Antifungal susceptibility testing should be determined before therapy to avoid treatment failures.

  16. Issues in identifying germ tube positive yeasts by conventional methods.

    Science.gov (United States)

    Yazdanpanah, Atta; Khaithir, Tzar Mohd Nizam

    2014-01-01

    Candida speciation is vital for epidemiology and management of candidiasis. Nonmolecular conventional methods often fail to identify closely related germ tube positive yeasts from clinical specimens. The present study was conducted to identify these yeasts and to highlight issues in conventional versus molecular methods of identification. A total of 98 germ tube positive yeasts from high vaginal swabs were studied over a 12-month period. Isolates were examined with various methods including growth at 42 °C and 45 °C on Sabouraud dextrose agar (SDA), color development on CHROMagar Candida medium, chlamydospore production on corn meal agar at 25 °C, carbohydrate assimilation using ID 32C system, and polymerase chain reaction using a single pair of primers targeting the hyphal wall protein 1 (Hwp1) gene. Of all the isolates studied, 97 were molecularly confirmed as C. albicans and one isolate was identified as C. dubliniensis. No C. africana was detected in this study. The molecular method used in our study was an accurate and useful tool for discriminating C. albicans, C. dubliniensis, and C. africana. The conventional methods, however, were less accurate and riddled with many issues that will be discussed in further details. © 2013 Wiley Periodicals, Inc.

  17. Studies on the biological effects of deuteriated organic compounds

    International Nuclear Information System (INIS)

    Hellgren, L.; Liljemark, A.; Vincent, J.

    1976-01-01

    In order to visualise the morphological changes of Epidermophyton floccosum associated with exposure to perdeuteriated n-hendecanoic acid, the architecture of the dermatophyte was investigated by means of interference contrast and scanning electron microscopy. The mortphology of mycelia grown on substrate containing perdeuteriated n-hendecnoic acid, or the unlabelled analogue, was compared. The perdeuteriated n-hendecanoic acid produced a characteristic undulant effect of the hyphae. The characteristic wave-lake appearance of the mycelia looked similar to the curling effect occurring after treatment of dermatophytes with griseofulvin, but was not so pronounced. Perdeuteriated n-hendecanoic acid, unlike the unlabelled analogue, also seems to cause a reduction of the number of chlamydospores perforations of the macroconidia. The changes in the morphological structure of Epidermophyton floccosum exposed to perdeuteriated n-hendecanoic acid have been investigated. Morphological examination of mycelia exposed to this substance by interference contrast microscopy demonstrated a picture of defect hyphae and macroconidia. By the aid of scanning electron microscopy an attempt was made to obtain a better visualization of these changes at ultrastructural level

  18. CULTURE DESCRIPTION OF SOME SPONTANEOUS LIGNICOLOUS MACROMYCETES SPECIES

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    BALAEŞ TIBERIUS

    2012-12-01

    Full Text Available 24 species of lignicolous macromycetes from 4 taxonomic families and 2 orders, Class Agaricomycetes, Phyllum Basidiomycota, have been analyzed. The cultural characters of these isolates had been observed, some of them being little studied till now. The dikaryotic mycelium from the trama of the sporoms was used for the isolation purpose. The fungal isolates were cultivated onto malt extract-agar media (malt extract 20g l-1 and incubated at 25 °C, in the dark, for 6 weeks. The cultures were observed directly and using a Nikon stereomicroscope in order to measure the growth rhythm and to observe the changes of the colonies: edge, surface, reverse, shape, colour, smell, presence or absence of the exudates. After 6 weeks from the inoculation, microscopic slides were made in order to investigate the types of hyphae, the colour and the structure of the mycelium and to note the presence of particular elements: cuticle, chlamydospors, arthrospores, conidia, and basidia. We noticed that the analyzed species present similar characters but also significant differences between them.

  19. The usefulness of DNA sequencing after extraction by Whatman FTA filter matrix technology and phenotypic tests for differentiation of Candida albicans and Candida dubliniensis.

    Science.gov (United States)

    Kiraz, Nuri; Oz, Yasemin; Aslan, Huseyin; Muslumanoglu, Hamza

    2014-02-01

    Since C. dubliniensis is similar to C. albicans phenotypically, it can be misidentified as C. albicans. We aimed to investigate the prevalence of C. dubliniensis among isolates previously identified as C. albicans in our stocks and to compare the phenotypic methods and DNA sequencing of D1/D2 region on the ribosomal large subunit (rLSU) gene. A total of 850 isolates included in this study. Phenotypic identification was performed based on germ tube formation, chlamydospore production, colony colors on chromogenic agar, inability of growth at 45 °C and growth on hypertonic Sabouraud dextrose agar. Eighty isolates compatible with C. dubliniensis by at least one phenotypic test were included in the sequence analysis. Nested PCR amplification of D1/D2 region of the rLSU gene was performed after the fungal DNA extraction by Whatman FTA filter paper technology. The sequencing analysis of PCR products carried out by an automated capillary gel electrophoresis device. The rate of C. dubliniensis was 2.35 % (n = 20) among isolates previously described as C. albicans. Consequently, none of the phenotypic tests provided satisfactory performance alone in our study, and molecular methods required special equipment and high cost. Thus, at least two phenotypic methods can be used for identification of C. dubliniensis, and molecular methods can be used for confirmation.

  20. Meloidogyne javanica control by Pochonia chlamydosporia, Gracilibacillus dipsosauri and soil conditioner in tomato

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    Guilherme Silva de Podestá

    2013-06-01

    Full Text Available Organic matter plays a fundamental role in the antagonistic activity of microorganisms against phytonematode populations on the soil. In this study, the compatibility between the fungus Pochonia chlamydosporia (Pc-12 and the rhizobacterium Gracilibacillus dipsosauri (MIC 14 was evaluated in vitro, as well as the effect of the fungus at the concentration of 5,000 chlamydospores per gram of soil, rhizobacterium at 4.65 x 10(9 cells/g of soil, and the soil conditioner Ribumin® at 10 g/pot, either alone or in combination, against Meloidogyne javanica population in tomato plants (3,000 eggs/pot. A suspension of water or Ribumin® alone was applied on the soil as negative control, while a suspension of nematode eggs was applied as positive control. The reduction in the number of galls in roots per plant was 48 and 41% for the treatments Ribumin + MIC 14 + Pc-12 and MIC 14 + Pc-12, respectively. Regarding to the number of eggs per plant, MIC 14 and Pc-12 + Ribumin led to a reduction by 26 and 21%, respectively, compared to the control treatment. Interaction between the nematophagous fungus and the rhizobacterium was positive for the nematode control, even though G. dipsosauri inhibited P. chlamydosporia growth by up to 30% in in vitro tests.

  1. Granular formulation of Fusarium oxysporum for biological control of faba bean and tomato Orobanche.

    Science.gov (United States)

    Nemat Alla, Mamdouh M; Shabana, Yasser M; Serag, Mamdouh M; Hassan, Nemat M; El-Hawary, Mohamed M

    2008-12-01

    Orobanche spp. represent a serious threat to a wide range of crops. They are difficult targets for herbicides, and biological control could provide a possible solution. This work therefore aimed to formulate mycoherbicides of Fusarium with adequate shelf life and virulence against Orobanche but safe to faba bean and tomato. Only two isolates of Fusarium oxysporum Schlecht. (Foxy I and Foxy II) obtained from diseased Orobanche shoots were found to be pathogenic to Orobanche crenata Forsk. and Orobanche ramosa L. Conidial suspension of both isolates significantly decreased germination, attachments and tubercles of Orobanche. Microconidia and chlamydospores of both isolates were formulated as mycoherbicides encapsulated in a wheat flour-kaolin matrix (four different formulations). All formulations greatly diminished Orobanche emerged shoots, total shoot number, shoot height, attachment of emerged shoots, the germinated seeds that succeeded in emerging above the soil surface and dry weight. Meanwhile, disease incidence and disease severity of emerged shoots were enhanced. The shelf life was adequate, particularly for coarse, freshly prepared, low-temperature-stored, microconidia-rich formulations. The induced growth reduction of Orobanche-infected host plants seemed to be nullified by formulations, particularly at the highest dose. These formulations seemed to destroy Orobanche but appeared harmless to host plants. Hence, they could be efficiently used as mycoherbicides for biological control of Orobanche in faba bean and tomato.

  2. Production of pigment-free pullulan by swollen cell in Aureobasidium pullulans NG which cell differentiation was affected by pH and nutrition.

    Science.gov (United States)

    Li, Bing-xue; Zhang, Ning; Peng, Qing; Yin, Tie; Guan, Fei-fei; Wang, Gui-li; Li, Ying

    2009-08-01

    A black yeast strain "NG" was isolated from strawberry fruit and identified as Aureobasidium pullulans. Strain NG displayed yeast-like cell (YL), swollen cell (SC), septate swollen cell (SSC), meristematic structure (MS), and chlamydospore (CH) morphologies. pH was the key factor regulating cell morphogenesis of strain NG. Differentiation of YL controlled by extracellular pH had no relationship with nutrition level. YL was maintained at pH >6.0, but was transformed into SC at pH approximately 4.5. SC, a stable cell type of A. pullulans, could bud, septate, or transform into MS or CH, in response to nutrition level and low pH. SC produced swollen cell blastospores (SCB) at pH 2.1 with abundant nutrition, and could transform into MS at lower pH (1.5). SC was induced to form CH by low level nutrition and pH melanin) were produced by SC of strain NG. Pullulan content of the polysaccharides was very high (98.37%). Fourier-transform infrared spectroscopy confirmed that chemical structures of the polysaccharides and standard pullulan were identical. Swollen cells produced 2.08 mg/ml non-pigmented polysaccharides at 96 h in YPD medium. Controlling pH of fermentation is an effective and convenient method to harvest SC for melanin-free pullulan production.

  3. Comparison of duplex PCR and phenotypic analysis in differentiating Candida dubliniensis from Candida albicans from oral samples.

    Science.gov (United States)

    Sampath, Asanga; Weerasekera, Manjula; Dilhari, Ayomi; Gunasekara, Chinthika; Bulugahapitiya, Uditha; Fernando, Neluka; Samaranayake, Lakshman

    2017-12-01

    Candida dubliniensis shares a wide range of phenotypic characteristics with Candida albicans including a common trait called germ tube positivity. Hence, laboratory differentiation of these two species is cumbersome. Duplex PCR analyses for C. albicans and C. dubliniensis was performed directly on DNA extracted from a total of 122 germ tube positive isolates derived from 100 concentrated oral rinse samples from a random cohort of diabetics attending a clinic in Sri Lanka. These results were confirmed by DNA sequencing of internal transcribed spacer (ITS) region of rDNA of the yeasts. Performance efficacy of duplex PCR was then compared with phenotypic identification using a standard battery of phenotypic tests. Of the 122 germ tube positive isolates three were identified by duplex PCR as C. dubliniensis and the remainder as C. albicans. On the contrary, when the standard phenotypic tests, sugar assimilation and chlamydospore formation, were used to differentiate the two species 13 germ tube positive isolates were erroneously identified as C. dubliniensis. Duplex PCR was found to be rapid, sensitive and more specific than phenotypic identification methods in discriminating C. dubliniensis from C. albicans. This is also the first report on the oral carriage of C. dubliniensis in a Sri Lankan population.

  4. Identification of Yeast Species In the Oral Cavity of Iranian Soldiers By Disk Diffusion Method

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    M. Imami

    2008-02-01

    Full Text Available Background:The disk diffusion method for identification of yeasts species was performed based on different but distinct susceptibilities of yeasts spp.to chemicals:janus green, ethidium bromide,2,3,5-triphenyltetrazolium chloride, brilliant green, cycloheximide and rhodamine 6G. Methods: Atotal of 568 Iranian soldiers went under study for isolation and identification of Yeast species from their oral cavity. Asterile swab was used for each individual and specimens were collected from the nasopharynx region, then inoculated to petri dishes containing Sabouraud Dextrose Agar and incubated for 48 hrs at 37 °C. All colonies were counted and stocked in distilled water and stored in a refrigerator for further analysis. The yeasts were identified by the “disk diffusion test” [6,8]. This is a simple, rapid, accurate, and inexpensive technique presented by Sobczak [8]. By this method we identified yeast species within 24-48 hrs. Results: 51.4% of petri dishes were positive for yeast species and 318 strains were identified. Candida albicans, Candida kefyr, Candida tropicalis and Candida guilliermondii were the most common yeast species isolated from the oral cavity of soldiers. Conclusion: We used this method because of its simplicity and other beneficial characteristics for rapid identification of large and numerous isolates and the results were compared with other morphological characters such as chlamydospore and germ tube production. In addition,we used some type strains (Candida parapsilosis: PTCC 5089,Candida tropicalis: PTCC 5028,Saccharomyces cerevisiae:PTCC 5052,Candida lipolytica: PTCC 5063,Candida lipolytica:PTCC 5064,and the results were acceptable.

  5. Characterization of Candida species isolated from cases of lower respiratory tract infection.

    Science.gov (United States)

    Jha, B J; Dey, S; Tamang, M D; Joshy, M E; Shivananda, P G; Brahmadatan, K N

    2006-01-01

    (1) To identify and characterize the Candida species isolates from lower respiratory tract infection. (2) to determine the rate of isolation of Candida species from sputum samples. This study was carried out in the Department of Microbiology, Manipal Teaching Hospital, Pokhara, Nepal from June 2002 to January 2003. A total of 462 sputum samples were collected from patients suspected lower respiratory tract infection. The samples were processed as Gram staining to find out the suitability of the specimen, cultured on Sabouraud's Dextrose Agar (SDA) and also on blood agar and chocolate agar to identify the potential lower respiratory tract pathogens. For the identification of Candida, sputum samples were processed for Gram stain, culture, germ tube test, production of chlamydospore, sugar fermentation and assimilation test. For the identification of bacteria, Gram stain, culture, and biochemical tests were performed by standardized procedure. Out of 462 samples, 246 (53.24%) samples grew potential pathogens of lower respiratory tract. Among them Haemophilus influenzae 61(24.79%) and Streptococcus pneumoniae 57 (23.17%) were the predominant bacterial pathogens. Candida species were isolated from 30 samples (12.2%). The majority of Candida species amongst the Candida isolates were Candida albicans 21(70%) followed by Candida tropicalis 4(13.33%). Candida krusei 3(10%), Candida parapsilosis 1(3.33%) and Candida stellatoidea 1(3.33%). The highest rate of isolation of Candida was between the age of 71 and 80. Candida isolation from sputum samples is important as found in the present study in which Candida species were the third most common pathogen isolated from patients with lower respiratory tract infection.

  6. Bmh1p (14-3-3) mediates pathways associated with virulence in Candida albicans.

    Science.gov (United States)

    Kelly, Michelle N; Johnston, Douglas A; Peel, Bethany A; Morgan, Timothy W; Palmer, Glen E; Sturtevant, Joy E

    2009-05-01

    The ability of the pathogenic fungus Candida albicans to cause disease requires rapid adaptation to changes in the host environment and to an evolving host immune response. The identification of 'virulence factors' using in vitro characterization of mutant strains has traditionally relied on a common set of phenotypic and biochemical assays (most often performed at 30 degrees C) and the subsequent correlation with their corresponding virulence in mouse models of disease. Utilizing a panel of isogenic mutants for the multifunctional signal-modulating 14-3-3 protein (Bmh1p), we have found that specific mutations affect a variety of different pathways currently associated with virulence, including those involved with the formation of filaments, as well as interaction with host immune cells. Surprisingly, our studies revealed that deficiencies in many of these pathways do not always correlate with virulence in a mouse model of disseminated infection. Mutations within the binding pocket of Bmh1p that affect the ability of the protein to efficiently bind ligand had varying effects on the results of a number of in vitro and in vivo assays. The capability, in vitro, to filament in embedment conditions, and to filament and form chlamydospores under microaerophilic conditions on cornmeal agar, does not correlate with virulence. It is likely that only a subset of hyphal signalling pathways is actually required for the establishment of infection in the disseminated mouse model. Most importantly, our results suggest that the delayed onset of log-phase [corrected] growth in vitro at 37 degrees C, and not at 30 degrees C, results in an inability of these mutants to rapidly adjust to environmental changes in vivo and may be responsible for their increased clearance and reduced virulence. It is critical, therefore, that future in vitro studies of putative virulence factors in C. albicans include careful characterization at physiological temperatures.

  7. Fungicidal activity of Eucalyptus tereticornis essential oil on the pathogenic fungus Fusarium oxysporum Actividad antimicótica del aceite esencial a partir de Eucalyptus tereticornis sobre el hongo patógeno Fusarium oxysporum

    Directory of Open Access Journals (Sweden)

    Walter Murillo Arango

    2011-06-01

    Full Text Available The objective of present paper was to determine the antifungal activity of the Eucalyptus tereticornis (Myrtaceae essential oil and two fractions on the Fusarium oxysporum mushroom, a pathogen with clinical and agricultural significance. The total citronelal (44.8 % and geraniol (9.78 % essential oil had a fungicidal effect at a 3 g/L concentration and a fungicidal activity at small concentrations. The A and B fractions composed most of p-mentane-3,8-diol (18.95 % and geraniol acetate (24.34 %, respectively were more active than the total extract. The observations at microscopic level showed damages and changes in hyphae and chlamydospores, as well as a decrease in the number of conidia. The observed fungicidal activity and the morphologic damages were dependent on the concentration.El objetivo de este trabajo fue determinar la actividad antifúngica del aceite esencial de Eucalyptus tereticornis (Myrtaceae y 2 fracciones sobre el hongo Fusarium oxysporum, patógeno de importancia tanto clínica como agrícola. El aceite esencial total, compuesto principalmente por citronelal (44,8 %, citronelol (9,78 % presentó un efecto fungicida a una concentración de 3 g/L y actividad fungistática a concentraciones menores. La fracciones A y B compuestas en su mayoría por p-mentano-3,8-diol (18,95 % y acetato de citronelol (24,34 % respectivamente fueron más activas que el extracto total. Las observaciones a nivel microscópico mostraron daños y cambios en hifas y clamidosporas, así como disminución en el número de conidias. La actividad fungistática observada y los daños morfológicos fueron dependientes de la concentración.

  8. The antibiotic polymyxin B exhibits novel antifungal activity against Fusarium species.

    Science.gov (United States)

    Hsu, Li-Hang; Wang, Hsuan-Fu; Sun, Pei-Lun; Hu, Fung-Rong; Chen, Ying-Lien

    2017-06-01

    The genus Fusarium comprises many species, including Fusarium oxysporum, Fusarium solani, Fusarium graminearum and Fusarium verticillioides, and causes severe infections in plants and humans. In clinical settings, Fusarium is the third most frequent mould to cause invasive fungal infections after Aspergillus and the Mucorales. F. solani and F. oxysporum are the most prevalent Fusarium spp. causing clinical disease. However, few effective antifungal drugs are available to treat human and plant Fusarium infections. The cationic peptide antibiotic polymyxin B (PMB) exhibits antifungal activity against the human fungal pathogens Candida albicans and Cryptococcus neoformans, but its efficacy against Fusarium spp. is unknown. In this study, the antifungal activity of PMB was tested against 12 Fusarium strains that infect humans and plants (banana, tomato, melon, pea, wheat and maize). PMB was fungicidal against all 12 Fusarium strains, with minimum fungicidal concentrations of 32 µg/mL or 64 µg/mL for most strains tested, as evidenced by broth dilution, methylene blue staining and XTT reduction assays. PMB can reduce the germination rates of conidia, but not chlamydospores, and can cause defects in cell membrane integrity in Fusarium strains. PMB exhibits synergistic activity with posaconazole and can potentiate the effect of fluconazole, voriconazole or amphotericin B against Fusarium spp. However, PMB does not show synergistic effects with fluconazole against Fusarium spp. as it does against Candida glabrata and C. neoformans, indicating evolutionary divergence of mechanisms between yeast pathogens and the filamentous fungus Fusarium. Copyright © 2017 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

  9. Ijuhya vitellina sp. nov., a novel source for chaetoglobosin A, is a destructive parasite of the cereal cyst nematode Heterodera filipjevi.

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    Samad Ashrafi

    Full Text Available Cyst nematodes are globally important pathogens in agriculture. Their sedentary lifestyle and long-term association with the roots of host plants render cyst nematodes especially good targets for attack by parasitic fungi. In this context fungi were specifically isolated from nematode eggs of the cereal cyst nematode Heterodera filipjevi. Here, Ijuhya vitellina (Ascomycota, Hypocreales, Bionectriaceae, encountered in wheat fields in Turkey, is newly described on the basis of phylogenetic analyses, morphological characters and life-style related inferences. The species destructively parasitises eggs inside cysts of H. filipjevi. The parasitism was reproduced in in vitro studies. Infected eggs were found to harbour microsclerotia produced by I. vitellina that resemble long-term survival structures also known from other ascomycetes. Microsclerotia were also formed by this species in pure cultures obtained from both, solitarily isolated infected eggs obtained from fields and artificially infected eggs. Hyphae penetrating the eggshell colonised the interior of eggs and became transformed into multicellular, chlamydospore-like structures that developed into microsclerotia. When isolated on artificial media, microsclerotia germinated to produce multiple emerging hyphae. The specific nature of morphological structures produced by I. vitellina inside nematode eggs is interpreted as a unique mode of interaction allowing long-term survival of the fungus inside nematode cysts that are known to survive periods of drought or other harsh environmental conditions. Generic classification of the new species is based on molecular phylogenetic inferences using five different gene regions. I. vitellina is the only species of the genus known to parasitise nematodes and produce microsclerotia. Metabolomic analyses revealed that within the Ijuhya species studied here, only I. vitellina produces chaetoglobosin A and its derivate 19-O-acetylchaetoglobosin A. Nematicidal

  10. Ijuhya vitellina sp. nov., a novel source for chaetoglobosin A, is a destructive parasite of the cereal cyst nematode Heterodera filipjevi.

    Science.gov (United States)

    Ashrafi, Samad; Helaly, Soleiman; Schroers, Hans-Josef; Stadler, Marc; Richert-Poeggeler, Katja R; Dababat, Abdelfattah A; Maier, Wolfgang

    2017-01-01

    Cyst nematodes are globally important pathogens in agriculture. Their sedentary lifestyle and long-term association with the roots of host plants render cyst nematodes especially good targets for attack by parasitic fungi. In this context fungi were specifically isolated from nematode eggs of the cereal cyst nematode Heterodera filipjevi. Here, Ijuhya vitellina (Ascomycota, Hypocreales, Bionectriaceae), encountered in wheat fields in Turkey, is newly described on the basis of phylogenetic analyses, morphological characters and life-style related inferences. The species destructively parasitises eggs inside cysts of H. filipjevi. The parasitism was reproduced in in vitro studies. Infected eggs were found to harbour microsclerotia produced by I. vitellina that resemble long-term survival structures also known from other ascomycetes. Microsclerotia were also formed by this species in pure cultures obtained from both, solitarily isolated infected eggs obtained from fields and artificially infected eggs. Hyphae penetrating the eggshell colonised the interior of eggs and became transformed into multicellular, chlamydospore-like structures that developed into microsclerotia. When isolated on artificial media, microsclerotia germinated to produce multiple emerging hyphae. The specific nature of morphological structures produced by I. vitellina inside nematode eggs is interpreted as a unique mode of interaction allowing long-term survival of the fungus inside nematode cysts that are known to survive periods of drought or other harsh environmental conditions. Generic classification of the new species is based on molecular phylogenetic inferences using five different gene regions. I. vitellina is the only species of the genus known to parasitise nematodes and produce microsclerotia. Metabolomic analyses revealed that within the Ijuhya species studied here, only I. vitellina produces chaetoglobosin A and its derivate 19-O-acetylchaetoglobosin A. Nematicidal and nematode

  11. The combined use of Pochonia chlamydosporia and plant defence activators - a potential sustainable control strategy for Meloidogyne chitwoodi

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    Maria Clara VIEIRA DOS SANTOS

    2014-05-01

    Full Text Available Sustainable strategies are required for control of the root-knot nematode Meloidogyne chitwoodi to reduce dependence on toxic chemical pesticides. The efficacy of the nematophagous fungus Pochonia chlamydosporia in biocontrol could be enhanced by integration with control measures that reduce initial nematode infestations. The use of foliar sprays with plant defence activators can reduce the susceptibility of potato plants to M. chitwoodi. This study assessed effects of combined soil application of P. chlamydosporia with foliar sprays of benzothiadiazole (BTH or cis-jasmone on infection of potatoes by M. chitwoodi. Solanum tuberosum, cv. Désirée plants were grown in soil mixed with 5000 chlamydospores g-1 of soil, sprayed twice with BTH or cis-jasmone and inoculated with 300 M. chitwoodi second-stage juveniles. Forty-five days after inoculation, nematode reproduction, numbers of colony-forming units of the fungus g-1 of soil and g-1 of root, and egg parasitism were assessed by standard techniques. Foliar sprays of BTH or cis-jasmone combined with the fungus reduced nematode reproduction (P<0.05, LSD. The presence of the fungus slightly increased the efficacy of cis-jasmone, as the number of eggs per egg mass was less in plants treated both with cis-jasmone and the fungus than in the plants treated only with the defence activator. The proportion of parasitized eggs was greater in the cis-jasmone treatment where rhizosphere colonisation was less, suggesting that P. chlamydosporia became a poorer rhizosphere coloniser but a more efficient nematode parasite. The addition of P. chlamydosporia to soil in combination with application of inducers of plant defence could be an alternative control strategy to be used against M. chitwoodi in potato.

  12. Chitosan Increases Tomato Root Colonization by Pochonia chlamydosporia and Their Combination Reduces Root-Knot Nematode Damage

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    Nuria Escudero

    2017-09-01

    Full Text Available The use of biological control agents could be a non-chemical alternative for management of Meloidogyne spp. [root-knot nematodes (RKN], the most damaging plant-parasitic nematodes for horticultural crops worldwide. Pochonia chlamydosporia is a fungal parasite of RKN eggs that can colonize endophytically roots of several cultivated plant species, but in field applications the fungus shows a low persistence and efficiency in RKN management. The combined use of P. chlamydosporia with an enhancer could help its ability to develop in soil and colonize roots, thereby increasing its efficiency against nematodes. Previous work has shown that chitosan enhances P. chlamydosporia sporulation and production of extracellular enzymes, as well as nematode egg parasitism in laboratory bioassays. This work shows that chitosan at low concentrations (up to 0.1 mg ml-1 do not affect the viability and germination of P. chlamydosporia chlamydospores and improves mycelial growth respect to treatments without chitosan. Tomato plants irrigated with chitosan (same dose limit increased root weight and length after 30 days. Chitosan irrigation increased dry shoot and fresh root weight of tomato plants inoculated with Meloidogyne javanica, root length when they were inoculated with P. chlamydosporia, and dry shoot weight of plants inoculated with both P. chlamydosporia and M. javanica. Chitosan irrigation significantly enhanced root colonization by P. chlamydosporia, but neither nematode infection per plant nor fungal egg parasitism was affected. Tomato plants cultivated in a mid-suppressive (29.3 ± 4.7% RKN egg infection non-sterilized clay loam soil and irrigated with chitosan had enhanced shoot growth, reduced RKN multiplication, and disease severity. Chitosan irrigation in a highly suppressive (73.7 ± 2.6% RKN egg infection sterilized-sandy loam soil reduced RKN multiplication in tomato. However, chitosan did not affect disease severity or plant growth irrespective of

  13. Research on Candida dubliniensis in a Brazilian yeast collection obtained from cardiac transplant, tuberculosis, and HIV-positive patients, and evaluation of phenotypic tests using agar screening methods.

    Science.gov (United States)

    Ribeiro, Patrícia Monteiro; Querido, Silvia Maria Rodrigues; Back-Brito, Graziela Nueremberg; Mota, Adolfo José; Koga-Ito, Cristiane Yumi; Jorge, Antonio Olavo Cardoso

    2011-09-01

    The aim of this study was to research Candida dubliniensis among isolates present in a Brazilian yeast collection and to evaluate the main phenotypic methods for discrimination between C. albicans and C. dubliniensis from oral cavity. A total of 200 isolates, presumptively identified as C. albicans or C. dubliniensis obtained from heart transplant patients under immunosuppressive therapy, tuberculosis patients under antibiotic therapy, HIV-positive patients under antiretroviral therapy, and healthy subjects, were analyzed using the following phenotypic tests: formation and structural arrangement of chlamydospores on corn meal agar, casein agar, tobacco agar, and sunflower seed agar; growth at 45 °C; and germ tube formation. All strains were analyzed by polymerase chain reaction (PCR). In a preliminary screen for C. dubliniensis, 48 of the 200 isolates on corn meal agar, 30 of the 200 on casein agar, 16 of the 200 on tobacco agar, and 15 of the 200 on sunflower seed agar produced chlamydoconidia; 27 of the 200 isolates showed no or poor growth at 45 °C. All isolates were positive for germ tube formation. These isolates were considered suggestive of C. dubliniensis. All of them were subjected to PCR analysis using C. dubliniensis-specific primers. C. dubliniensis isolates were not found. C. dubliniensis isolates were not recovered in this study done with immunocompromised patients. Sunflower seed agar was the medium with the smallest number of isolates of C. albicans suggestive of C. dubliniensis. None of the phenotypic methods was 100% effective for discrimination between C. albicans and C. dubliniensis. Copyright © 2011 Elsevier Inc. All rights reserved.

  14. Identification of Candida dubliniensis in a study of HIV-seropositive pediatric dental patients.

    Science.gov (United States)

    Brown, D M; Jabra-Rizk, M A; Falkler, W A; Baqui, A A; Meiller, T F

    2000-01-01

    The combination of an immature immune system and suppressed cellular immunity in children with HIV infections provides optimal conditions for rapid disease progression. As a result, pediatric AIDS has become a major epidemiological challenge. Oral fungal colonization remains one of the most common opportunistic infections observed in both adult and pediatric HIV infected patients. Although Candida albicans is the most frequently isolated opportunistic fungal species, a recently characterized Candida species, C. dubliniensis, has gained considerable attention due to its almost exclusive association with HIV-seropositive individuals. The purpose of this study was to prospectively screen for the presence of C. dubliniensis among pediatric HIV+ patients. Oral samples taken from twenty-seven children were cultured for the presence of yeast. All positive yeast isolates obtained were screened for the presence of C. dubliniensis by use of tests for germ tube and chlamydospore production, detection of inability to grow at 45 degrees C, by colony color on CHROMagar Candida medium, coaggregation with Fusobacterium nucleatum ATCC 49256 and by the results of sugar assimilation testing with the API 20C AUX yeast identification system. Among the 27 patients tested, 3 patients were found to harbor C. dubliniensis, one of which also grew C. glabrata; 12 patients were colonized with C. albicans, while the remaining 12 patients were negative for yeast. Identification of the three C. dubliniensis isolates was genetically confirmed by electrophoretic karyotyping. All three C. dubliniensis isolates were found to be susceptible to fluconazole (MIC pediatric HIV seropositive population and support the need for further investigation into the prevalence and pathogenesis of C. dubliniensis.

  15. [Molecular epidemiology and antifungal susceptibility of Candida species isolated from urine samples of patients in intensive care unit].

    Science.gov (United States)

    Yüksekkaya, Serife; Fındık, Duygu; Arslan, Uğur

    2011-01-01

    The aims of this study were to analyse the amphotericin B and fluconazole susceptibility and molecular epidemiology of Candida strains (Candida albicans, Candida tropicalis and Candida glabrata) isolated from the urine samples of patients hospitalized in the intensive care unit. Identification of the isolates was done according to microscopic morphology (chlamydospor, blastospor, pseudohyphae and true hyphae) on cornmeal agar, germ tube formation and carbohydrate assimilation patterns (API ID 32C bioMérieux, France). Antifungal susceptibilities of the isolates were determined by in vitro broth microdilution method recommended by Clinical and Laboratory Standards Institute (CLSI). To investigate the clonal relationship of the isolates, randomly amplified polymorphic DNA (RAPD) analysis was performed by using Cnd3 primer. Of the 56 Candida isolates minimum inhibitory concentration (MIC) ranges, MIC50 and MIC90 values for amphotericin B were 0.125-1 µg/ml, 0.125 and 0.5 µg/ml for C.albicans, 0.125-1 µg/ml, 0.25 and 1 µg/ml for C.tropicalis and 0.125-1 µg/ml, 0.25 and 1 µg/ml for C.glabrata, respectively. Fluconazole MIC ranges, MIC50 and MIC90 values were 0.25-4 µg/ml, 0.25 and 0.5 µg/ml for C.albicans, 0.25-16 µg/ml, 0.5 and 1 µg/ml for C.tropicalis and 0.5-64 µg/ml, 8 and 16 µg/ml for C.glabrata, respectively. For amphotericin B, none of the isolates had high MIC values (MIC > 1 µg/ml). While one of the C.glabrata isolates was resistant to fluconazole (MIC ≥ 64 µg/ml), one C.tropicalis and two C.glabrata isolates were dose-dependent susceptible (MIC: 16-32 µg/ml). The results of RAPD analysis indicated an exogenous spread from two clones for C.albicans, one clone for C.glabrata and one clone for C.tropicalis. This study underlines the importance of molecular epidemiological analysis of clinical samples together with hospital environmental samples in terms of Candida spp. To determine the exogenous origin for the related strains and to prevent

  16. The role and relevance of phospholipase D1 during growth and dimorphism of Candida albicans.

    Science.gov (United States)

    Hube, B; Hess, D; Baker, C A; Schaller, M; Schäfer, W; Dolan, J W

    2001-04-01

    The phosphatidylcholine-specific phospholipase D1 (PLD1) in Saccharomyces cerevisiae is involved in vesicle transport and is essential for sporulation. The gene encoding the homologous phospholipase D1 from Candida albicans (PLD1) was used to study the role of PLD1 in this pathogenic fungus. In vitro and in vivo expression studies using Northern blots and reverse transcriptase-PCR showed low PLD1 mRNA levels in defined media supporting yeast growth and during experimental infection, while enhanced levels of PLD1 transcripts were detected during the yeast to hyphal transition. To study the relevance of PLD1 during yeast and hyphal growth, an essential part of the gene was deleted in both alleles of two isogenic strains. In vitro PLD1 activity assays showed that pld1 mutants produced no detectable levels of phosphatidic acid, the hydrolytic product of PLD1 activity, and strongly reduced levels of diacylglycerol, the product of lipid phosphate phosphohydrolase, suggesting no or a negligible background PLD1 activity in the pld1 mutants. The pld1 mutants showed no growth differences compared to the parental wild-type in liquid complex and minimal media, independent of the growth temperature. In addition, growth rates of pld1 mutants in media with protein as the sole source of nitrogen were similar to growth rates of the wild-type, indicating that secretion of proteinases was not reduced. Chlamydospore formation was normal in pld1 mutants. When germ tube formation was induced in liquid media, pld1 mutants showed similar rates of yeast to hyphal transition compared to the wild-type. However, no hyphae formation was observed on solid Spider medium, and cell growth on cornmeal/Tween 80 medium indicated aberrant morphogenesis. In addition, pld1 mutants growing on solid media had an attenuated ability to invade the agar. In a model of oral candidosis, pld1 mutants showed no attenuation of virulence. In contrast, the mutant was less virulent in two different mouse models

  17. Ocorrência de Fusarium solani f. sp. piperis em Piper nigrum no estado de Alagoas Report of Fusariom solani f. sp. piperis in Piper nigrum in the state of Alagoas

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    Juliana Paiva Carnaúba

    2007-03-01

    Full Text Available A pimenta-do-reino (Piper nigrum L. é uma planta trepadeira, pertencente à família Piperaceae. Ela é originária do Sudeste Asiático, sendo a mais comum e importante das especiarias. A fusariose, também conhecida por podridão do pé e podridão das raízes é a principal doença da cultura, de ocorrência restrita ao Brasil. Um isolado de Fusarium sp., encontrado infectando plantas de pimenta-do-reino cv. bragantina no município de União dos Palmares em Alagoas, foi caracterizado morfologicamente e teve sua patogenicidade confirmada em mudas deste hospedeiro. Os macroconídios apresentaram-se falcados, hialinos com três a cinco septos, com dimensões de 30,5 - 26,5 x 6,3 - 4,9 ìm, enquanto os microconídios apresentaram-se hialinos, unicelulares, elípticos ou alantóides medindo 16,6 - 4,9 x 6,5 - 3,3 ìm. Os clamidósporos foram abundantes em meio batata-dextrose-ágar. O isolado foi identificado como Fusarium solani f. sp. piperis Alb. tratando-se do primeiro relato deste patógeno em pimenta-do-reino no estado de Alagoas.The pepper-of the-kingdom (Piper nigrum L. it is a climbing plant, pertaining plant to the Piperaceae family. It is originary the southeastern Asian, being most common and important of the spices. Seedling death, also known for rottenness of the foot and rottenness of the root is the main illness of the culture, restricted occurrence to Brazil. Isolated of Fusarium sp., found contamined plants of bragantina pepper-do-kingdom cv. in the city of União dos Palmares in Alagoas, was characterized morphologycament and had its pathogenicity confirmed in changes of this host. The macroconidia slightly curved, typically canoe-shaped, hyaline with three the five septate, measuring 30,5 - 26,5 x 6,3 - 4,9 ìm. Microconidia hyaline, unicellulars, ellipticals or allantoises measuring 16,6 - 4,9 x 6,5 - 3,3 ìm. The chlamydospores had been abundant in half potato-dextrose-agar. The isolated one was identified as to Fusarium

  18. Drug resistance of yeasts isolated from oropharyngeal candidiasis in aids patients Resistência à drogas de leveduras isoladas de candidíase orofaríngea em pacientes com Aids

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    Maria do Rosário Rodrigues Silva

    1998-10-01

    Full Text Available Candida spp was isolated from 59 (68.60% out of eighty six samples of oral mucosa of AIDS patients. The identification, based or the production of a germ tube and chlamydospores, and on the assimilation and fermentation of carbohydrates, revealed 52 strains (88.13% of C. albicans, 4 (6.77% of C. tropicalis and 3 (5.08% of C. krusei. The susceptibility of these strains to amphotericin B, flucytosine, itraconazole, fluconazole and ketoconazole was determined using the agar dilution method. Comparing the minimum inhibitory concentration values found in the susceptibility test with the serum levels achieved by these drugs, only 8.47% and 5.08% of the yeasts strains proved to be resistant to amphotericin B and flucitosyne, respectively. A high frequency of strains resistant to azole derivatives (25.42%, to itraconazole, 45.76%, to ketoconazole and 66.10% to fluconazole was observed.Entre oitenta e seis amostras da mucosa oral de pacientes com AIDS, 59 (68,60% foram positivas para leveduras do gênero Candida. A identificação, feita pela produção de tubo germinativo e clamidósporos e através de assimilação e fermentação de hidratos de carbono, revelou 52 cepas (88,13% de C.albicans, 4 (6,77% de C. tropicalis e 3 (5,08% de C.krusei. Avaliação destas leveduras para susceptibilidade in vitro frente a anfotericina B, flucitosina, itraconazol, fluconazol e cetoconazol, foi realizada pelo método de diluição em ágar. Comparando-se os valores de concentração inibitória mínima encontrados com os níveis séricos alcançados por estes antifúngicos verificou-se que apenas 8,47% e 5,08% das 59 leveduras foram resistentes a anfotericina B e flucitosina, respectivamente. Foi registrada uma percentagem de cepas resistentes aos derivados azólicos, sendo 25,42% ao itraconazol, 45,76% ao cetoconazol e 66,10% ao fluconazol.

  19. In-vitro predatory activity of nematophagous fungi from Costa Rica with potential use for controlling sheep and goat parasitic nematodes

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    Natalia Soto-Barrientos

    2011-03-01

    Full Text Available In tropical and subtropical regions of the world, parasitic diseases are a main cause of losses in livestock productivity. The increased acquired resistence to anthelmintics by gastrointestinal nematodes, requires biological control be considered as a potential feasible and effective alternative. The most effective natural soil enemies of nematodes are nematophagous fungi. In order to collect and identify predator nematophagous fungi (PNF, samples were obtained from 51 farms distributed throughout the seven provinces of Costa Rica. The origin samples included: soil from different crops (potatoes, tomatoes, bananas, ornamental plants, squash and coffee; animal feces (cattle, sheep, goat and horse; soil and fallen leaves from forest; and plants with signs of nematode infection. Each sample was processed using three techniques for the extraction of fungi from soil: sprinkling technique, soil dilution and humidity chamber. Twenty four strains of nematophagous fungi were found in 19 farms; 83.3% of the fungi were isolated by sprinkling technique. The following fungi were idenified: Arthrobotrys oligospora (n=13; Candelabrella musiformis (n=9; and for the first time there was isolation of A. conoides (n=1 and A. dactyloides (n=1 in the country. Moreover, 16 strains from Trichoderma (n=13, Beauveria (n=1, Clonostachys (n=1 and Lecanicillium (n=1 were obtained. In addition, pH of each possible fungal isolation source was measured, and it varied from 5.2 to 9.9, however PNF isolates fell within the range of 5.6 to 7.5. The PNF strains were cultivated in four different media for the production of chhlamydospores: potato dextrose agar (PDA; corn meal agar (CMA; malt extract agar (MEA and potato carrot agar (PCA. Out of these cultures, 95.8% of the strains formed chlamydospores primarily in the PCA. Of these strains, the profilic spore producers were subjected to ruminant artificial gastrointestinal conditions. A total of 14 fungi were tested, out of which

  20. In vitro susceptibility of Candida albicans clinical isolates to eight antifungal agents in Ouagadougou (Burkina Faso).

    Science.gov (United States)

    Zida, A; Yacouba, A; Bamba, S; Sangare, I; Sawadogo, M; Guiguemde, T; Kone, S; Traore, L K; Ouedraogo-Traore, R; Guiguemde, R T

    2017-12-01

    In recent years, the infection Candida albicans infection worldwide has risen, and the incidence of resistance to traditional antifungal therapies is also increasing. The aim of this study was to evaluate in vitro susceptibility of C. albicans clinical isolates to eight antifungal agents in Ouagadougou. A cross-sectional study was conducted from January 2013 to December 2015 at Yalgado Ouédraogo University Teaching Hospital. Two hundred seven strains have been isolated from 347 symptomatic patients received in different clinical services. Samples were cultured on Sabouraud Dextrose Agar supplemented with Cloramphenicol. Isolates were diagnosed as C. albicans using germ tube test, chlamydospore formation on Corn Meal Agar, and Api-Candida test (Biomérieux). Antifungal susceptibility testing was performed by disk diffusion method and isolates classified as susceptible, susceptible dose-dependent and resistant. Three hundred forty-seven (347) patients are included in this study. Two hundred and six (206) out of 347 collected samples (59.36%) were found positive for C. albicans. The strains were mostly isolated from vulvovaginal (49%) and oral infections (40.3%). The highest resistance rates of azoles were obtained with fluconazole (66.5%), itraconazole (52.3%) and ketoconazole (22.9%) when all clinical isolates were included. The resistance rates of fluconazole, itraconazole and ketoconazole remain highest for vulvovaginal and oral isolates. The rate of resistance to the polyene amphotericin B was 32.0% for all clinical isolates and was 56.4% for vulvovaginal strains. Resistance rate to nystatin was 6.3% for all clinical isolates. Cross-resistance analysis with data of all clinical strains revealed that the incidence of resistance to ketoconazole and itraconazole in fluconazole-resistant isolates was significantly higher than recorded for fluconazole-susceptible isolates. In vitro C. albicans antifungal susceptibility test in this study showed relatively high

  1. In-vitro predatory activity of nematophagous fungi from Costa Rica with potential use for controlling sheep and goat parasitic nematodes.

    Science.gov (United States)

    Soto-Barrientos, Natalia; de Oliveira, Jaqueline; Vega-Obando, Rommel; Montero-Caballero, Danilo; Vargas, Bernardo; Hernández-Gamboa, Jorge; Orozco-Solano, Claudio

    2011-03-01

    In tropical and subtropical regions of the world, parasitic diseases are a main cause of losses in livestock productivity. The increased acquired resistence to anthelmintics by gastrointestinal nematodes, requires biological control be considered as a potential feasible and effective alternative. The most effective natural soil enemies of nematodes are nematophagous fungi. In order to collect and identify predator nematophagous fungi (PNF), samples were obtained from 51 farms distributed throughout the seven provinces of Costa Rica. The origin samples included: soil from different crops (potatoes, tomatoes, bananas, ornamental plants, squash and coffee); animal feces (cattle, sheep, goat and horse); soil and fallen leaves from forest; and plants with signs of nematode infection. Each sample was processed using three techniques for the extraction of fungi from soil: sprinkling technique, soil dilution and humidity chamber. Twenty four strains of nematophagous fungi were found in 19 farms; 83.3% of the fungi were isolated by sprinkling technique. The following fungi were identified: Arthrobotrys oligospora (n = 13); Candelabrella musiformis (n = 9); and for the first time there was isolation of A. conoides (n = 1) and A. dactyloides (n = 1) in the country. Moreover, 16 strains from Trichoderma (n=13), Beauveria (n = 1), Clonostachys (n = 1) and Lecanicillium (n = 1) were obtained. In addition, pH of each possible fungal isolation source was measured, and it varied from 5.2 to 9.9, however PNF isolates fell within the range of 5.6 to 7.5. The PNF strains were cultivated in four different media for the production of chhlamydospores: potato dextrose agar (PDA); corn meal agar (CMA); malt extract agar (MEA) and potato carrot agar (PCA). Out of these cultures, 95.8% of the strains formed chlamydospores primarily in the PCA. Of these strains, the profilic spore producers were subjected to ruminant artificial gastrointestinal conditions. A total of 14 fungi were tested, out

  2. [Epidemiology of otomycoses at the University Hospital of Yopougon (Abidjan-Ivory Coast)].

    Science.gov (United States)

    Adoubryn, K D; N'Gattia, V K; Kouadio-Yapo, G C; Nigué, L; Zika, D K; Ouhon, J

    2014-06-01

    Otomycosis is a fungal infection, which leads to a damage of the external auditory meatus. The disease is worldwide in distribution but is said to be more common in tropical countries. Though otomycosis presumably occurs frequently in Africa, reports on its incidence and etiology are rare from Côte d'Ivoire. The objective of this study was to evaluate the prevalence of the disease and to identify aetiological agents as well as the risk factors. A cross-sectional study was carried out in the Otorhinolaryngology Department of the University Teaching Hospital of Yopougon from September 2007 to February 2008. For laboratory investigation, specimens were collected by means of a sterile swab. Samples were inoculated on Sabouraud's Dextrose Agar with and without antibiotics and incubated at 30°C for a period of 1 to 2 weeks. Identification was performed by direct microscopic examination on Cotton Blue Mount preparation and slide culture examination was used for differentiation of morphology. Biotyping was performed using Carbohydrate Fermentation tests, Carbohydrate Assimilation Tests (galerie Api 20 CAux TM - Sanofi Pasteur), Germ tube Test, detection of chlamydospore formation on corn meal agar. A total of 110 patients (sex-ratio=1.2) with suspected cases of otomycosis were investigated. Itching, otalgia, and hypoacusis were the symptoms reported by the patients and the apparent signs were debris in the ear, scabs and inflammation of the external auditory meatus. Of these, 88 cases (80%) were confirmed specifically of mycotic etiology on the basis of positive culture with 92 isolates consisting of yeasts (65.2%) and moulds (34.8%). The predominant etiological agents were Aspergillus flavus (28.4%), Candida guilliermondii (19.3%) and Candida parapsilosis (18.2%). The predisposing factors included previous otological pathology (P=0.010), frequent scratching of the external ear canal and use of ear drops (RR=3.47; IC 95%=1.3-9.27). This study revealed the great

  3. The epidemiology of Candida species associated with vulvovaginal candidiasis in an Iranian patient population.

    Science.gov (United States)

    Mahmoudi Rad, M; Zafarghandi, S; Abbasabadi, B; Tavallaee, M

    2011-04-01

    Vulvovaginal candidiasis is a common infection among women worldwide. According to previous epidemiological studies, Candida albicans is the most common species of Candida. The prevalence of non-Candida species, however, is increasing. Identification of Candida species among the population will not only help health professionals to choose suitable antifungal treatments, but also prevent development of drug resistance. The aim of this study was to identify, using chromogenic agar medium, the Candida species associated with vulvovaginal candidiasis among a sample of the Iranian population. In a prospective cohort study during a two year period from March 2006 to March 2008, swab samples of vaginal discharge/secretion were taken from 200 patients admitted to the gynecology clinic of Mahdieh Hospital (Tehran, Iran) with a clinical presentation suggestive of vulvovaginal candidiasis. The isolates obtained were cultured on Sabouraud dextrose agar and chromogenic agar medium. Candida species were also identified by germ tube formation in serum, chlamydospore production on Corn Meal Agar and carbohydrate absorption using the API 20C-AUX kit. Participants were asked to complete a questionnaire investigating the risk factors associated with candidiasis. An assessment of the different species of recurrent and non-recurrent candidiasis was also made. Descriptive statistics, chi-square test, and t-test were used to analyze the data. A total of 191 isolates were obtained from 175 vaginal specimens. Candida albicans accounted for 67% of the strains including single and mixed infections. The other identified species were Candida glabrata (18.3%), Candida tropicalis (6.8%), Candida krusei (5.8%), Candida parapsilosis (1.6%), and Candida guilliermondii (0.5%) respectively. Mixed infection with two or more species of Candida was seen in 10.3% of patients. The most common mixed cause was the combination of Candida albicans and Candida glabrata. Participants who were sexually active

  4. [Phenotypic and genotypic identification of Candida strains isolated as nosocomial pathogens].

    Science.gov (United States)

    Sahiner, Fatih; Ergünay, Koray; Ozyurt, Mustafa; Ardıç, Nurittin; Hoşbul, Tuğrul; Haznedaroğlu, Tunçer

    2011-07-01

    Over the last decade, there have been important changes in the epidemiology of Candida infections and antifungal agents used to treat these infections. In recent years, Candida species have emerged as important causes of invasive infections among patients in intensive care units. One of the main goals of this study was to evaluate the molecular epidemiology of infectious Candida species isolated in our hospital and accordingly supply data for hospital infection (HI) control. The other aim of this study was to evaluate effectiveness and practical applicability of traditional and molecular methods used to identify Candida isolates to the species level. A total of 77 Candida strains that were isolated from various clinical specimens of 60 hospitalized patients (29 male, 24 female; 7 were children) were included in the study. Fifty-seven (74%) of those isolates were defined as HI agents according to Centers for Disease Control and Prevention (CDC) criteria. The most common Candida species identified as agents of HI were C.albicans (22; 38.6%), followed by C.tropicalis (14; 24.6%), C.parapsilosis (13; 22.8%), C.glabrata (7; 12.3%) and Candida spp. (1; 1.75%). It was determined that bloodstream (26; 45.6%) and urinary tract infections (24; 42.1%) were the most frequently encountered nosocomial infections caused by Candida species. In addition it was detected that the most frequent causative agent of bloodstream infections was C.parapsilosis (10; 38.5%) and of urinary tract infections was C.albicans (12; 50%). The evaluation of advantages and disadvantages of traditional phenotypic methods [germ tube formation, chlamydospore formation in corn meal agar, growth at 45°C, colony characteristics on CHROMagar Candida medium, carbohydrate assimilation properties detected by API ID 32C (BioMerieux, France) system] and some molecular techniques [polymerase chain reaction (PCR) by using ITS-1, ITS-3 and ITS 4 primers, PCR-Restriction fragment length polymorphism (RFLP), PCRRFLP

  5. Comportamento anti-inflamatório da IL-6 nos derrames pleurais pós-revascularização do miocárdio*

    Directory of Open Access Journals (Sweden)

    António M.S. Chibante

    2007-05-01

    Full Text Available Resumo: Introdução: O comportamento pleural pós-cirurgia de revascularização do miocárdio (PCRM não está devidamente esclarecido em relação à resposta infla-matória local e requer maior interesse por ser uma observação constante e ainda pouco estudada.Objectivo: Avaliar o comportamento de algumas citocinas, em especial o possível papel anti-inflamatório da IL-6 (proteína envolvida na síntese da cortisona, no líquido pleural PCRM, uma vez que a sua actividade pró-inflamatória é constantemente referida, assim como o seu papel de citocina de fase de resposta aguda ao lado do TNF-α e da IL-1β nos processos inflamatórios agudos.Casuística e método: foram estudadas e analisadas pelo método ELISA as citocinas TNF-α, IL-1β, IL-2, IL-6, IL-8, VEGF e TGF-β em 16 transudatos e 43 exsudatos de líquido pleural em três tempos da fase aguda (2, 24 e 48 horas PCRM no Instituto do Coração e Serviço de Pneumologia da USP – Brasil.Resultados: Tanto o TNF-α como IL-2 não sofre-ram qualquer tipo de elevação dos seus níveis enquanto os da IL-1β só se elevaram a partir das 24 horas, o que coincidiu com a queda da curva da citocina anti-infla-matória TGF-β, que desde o início foi caindo flagran-temente até aos valores dos transudatos. A IL-8 perma-neceu elevada nas três fases e o VEGF foi ascendendo os seus níveis, que permaneceram estáveis nas 24 e 48 horas seguintes. A IL-6 mostrou-se em concentrações elevadas desde o início, apresentando-se como a úni-ca citocina com potencial anti-inflamatório durante as três fases de avaliação.Conclusões: Concluímos que a citocina IL-6 parece ter papel anti-inflamatório de destaque e superior ao TGF-β nos derrames pleurais PCRM e que o seu comportamento desarticula, pelo menos neste tipo de derrame, a ideia de citocina pr

  6. In-vitro predatory activity of nematophagous fungi from Costa Rica with potential use for controlling sheep and goat parasitic nematodes

    Directory of Open Access Journals (Sweden)

    Natalia Soto-Barrientos

    2011-03-01

    Full Text Available In tropical and subtropical regions of the world, parasitic diseases are a main cause of losses in livestock productivity. The increased acquired resistence to anthelmintics by gastrointestinal nematodes, requires biological control be considered as a potential feasible and effective alternative. The most effective natural soil enemies of nematodes are nematophagous fungi. In order to collect and identify predator nematophagous fungi (PNF, samples were obtained from 51 farms distributed throughout the seven provinces of Costa Rica. The origin samples included: soil from different crops (potatoes, tomatoes, bananas, ornamental plants, squash and coffee; animal feces (cattle, sheep, goat and horse; soil and fallen leaves from forest; and plants with signs of nematode infection. Each sample was processed using three techniques for the extraction of fungi from soil: sprinkling technique, soil dilution and humidity chamber. Twenty four strains of nematophagous fungi were found in 19 farms; 83.3% of the fungi were isolated by sprinkling technique. The following fungi were idenified: Arthrobotrys oligospora (n=13; Candelabrella musiformis (n=9; and for the first time there was isolation of A. conoides (n=1 and A. dactyloides (n=1 in the country. Moreover, 16 strains from Trichoderma (n=13, Beauveria (n=1, Clonostachys (n=1 and Lecanicillium (n=1 were obtained. In addition, pH of each possible fungal isolation source was measured, and it varied from 5.2 to 9.9, however PNF isolates fell within the range of 5.6 to 7.5. The PNF strains were cultivated in four different media for the production of chhlamydospores: potato dextrose agar (PDA; corn meal agar (CMA; malt extract agar (MEA and potato carrot agar (PCA. Out of these cultures, 95.8% of the strains formed chlamydospores primarily in the PCA. Of these strains, the profilic spore producers were subjected to ruminant artificial gastrointestinal conditions. A total of 14 fungi were tested, out of which

  7. Nail and skin lesions caused by scytalidium dimidiatum in Medellín (Colombia, 1990-1999: report of 128 cases and review of the name of the agent Lesiones ungueales y cutáneas por Scytalidium dimidiatum en Medellín (Colombia, 1990-1999. Presentación de 128 casos y revisión del problema del nombre del agente

    Directory of Open Access Journals (Sweden)

    Jaime Carmona Fonseca

    2000-03-01

    hyphae alone or with chlamydospores (21% and yeast cells alone or with pseudohyphae (5%. There was no significant difference between the results of nail and skin Objetivo:Revisar el problema de la denominación del agente y presentar los rasgos epidemiológicos de 128 casos de lesiones en uñas o en piel por Scytalidium dimidiatum. Diseño: Estudio descriptivo, en parte retrospectivo (1990-1996 y en parte prospectivo (1997-1999. Marco de referencia: hasta hace poco tiempo hubo problemas con la nomenclatura del agente, pero ahora existen los argumentos para despejar las dificultades y ese es el primer objetivo del trabajo: presentar esos argumentos e indicar la manera de usar bien uno u otro nombre. Por otra parte, la importancia del S. dimidiatum como causa de lesiones en humanos aumenta cada día. Este informe, restringido a lesiones en uñas y piel, recoge una cifra muy alta de pacientes (128 casos, correspondientes al lapso 1990-1999. Métodos: los autores revisaron cada uno de los registros de laboratorio y, aplicando criterios de consenso, resolvieron las dudas. Se utilizaron criterios estándar de identificación micológica. El análisis estadístico de los datos se hizo con el programa EpiInfo versión 6.04. y consistió, principalmente, en evaluar la asociación de variables con la chi cuadrada y en comparar posiciones de datos continuos y ordenados según su magnitud con la prueba H de Kruskal-Wallis. Resultados: los diferentes nombres del agente (Nattrassia mangiferae, Scytalidium dimidiatum, Scytalidium hialinum, Scytalidium lignicola, Hendersonula toruloidea deben usarse en situaciones precisas, que se describen aquí. Se estudiaron 128 muestras de las cuales se aisló por cultivo Scytalidium dimidiatum: 102 en uñas (92% en pies y 26 en lesiones extraungueales (73% interdigitales en pies. Las características epidemiológicas de un caso típico de lesión ungueal por S. dimidiatum indican que en el 41% de las veces es una mujer, de 21 a 49 años, que usa

  8. Researches about selecting resistant melon types to fusarium oxyporum f. sp.melonis race 1,2 by using tissue culture and mutation techniques

    International Nuclear Information System (INIS)

    2009-01-01

    Fusarium wilt is a vascular disease of the Cucurbitaceae family caused by the soil fungus Fusarium oxysporum f. sp. melonis (FOM), which is very detrimental to muskmelons (Cucumis melo L.). Fusarium wilt of melon is prevalent in temperate and tropical regions and causes a worldwide problem. FOM can survive in the soil for extended periods of time as chlamydospores, and is capable of colonizing crop residues and roots of most crops grown in rotation with melon. The only effective control is the use of resistant varieties. Four races of FOM have been identified, namely 0, 1, 2 and 1.2. Race 1.2 was further subdivided into race 1.2y and 1.2w, which cause yellowing and wilt symptoms, respectively. Two resistance genes (Fom-1 and Fom-2) have been identified in melons. Fom-1 confers resistance to FOM races 0 and 2, and Fom-2 confers resistance to races 0 and 1. These two genes are extensively used in breeding programmes, which can be assisted by marker assisted selection using markers linked to these resistance genes. No genes have been identified that confer resistance to race 1.2. However, polygenic recessive genes have been found to confer resistance to race 1.2 in Piboule genotypes. Melon production in Turkey is 1,700,000 tons and it is declining the year after year because of Fusarium wilt. Therefore, Fusarium wilt has a high economic importance in the cultivation of muskmelon in Turkey. In some parts of Turkey the prevalent races of this pathogen were determined. FOM has caused severe losses for farmers as our native cultivars are not resistant to this disease. It is believed our native cultivars will disappear if resistance to FOM is not introduced into the cultivated material. For this reason, many scientists in Turkey are focusing on research to develop new resistant cultivars via conventional and biotechnological breeding methods. In vitro techniques became widely spread during the 20th century, and their potential to make important contributions to plant