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Sample records for isolated invasive aspergillus

  1. Triazole Resistance Is Still Not Emerging in Aspergillus fumigatus Isolates Causing Invasive Aspergillosis in Brazilian Patients

    NARCIS (Netherlands)

    Negri, C.E.; Goncalves, S.S.; Sousa, A.C.P.; Bergamasco, M.D.; Martino, M.D.V.; Queiroz-Telles, F.; Aquino, V.R.; Castro, P.T.O.; Hagen, F.; Meis, J.F.G.M.; Colombo, A.L.

    2017-01-01

    Aspergillus fumigatus azole resistance has emerged as a global health problem. We evaluated the in vitro antifungal susceptibility of 221 clinical A. fumigatus isolates according to CLSI guidelines. Sixty-one isolates exhibiting MICs at the epidemiological cutoff value (ECV) for itraconazole or

  2. Liver injury in invasive aspergillus. Echographic findings

    International Nuclear Information System (INIS)

    Otero Fernandez, R.; Garcia Revillo, J.; Paez Moreno, J.; Zurera Tendero, L.J.

    1994-01-01

    Aspergillus is the second most common mycoses in immuno compromised patients. The invasive form is associated with a mortality of approximately 100%. We present a case of invasive aspergillus in a heart transplant recipient in whom ultrasound disclosed the presence of liver injury which was later confirmed by necropsy. We review the available literature. (Author) 15 refs

  3. Aspergillus felis sp nov., an Emerging Agent of Invasive Aspergillosis in Humans, Cats, and Dogs

    NARCIS (Netherlands)

    Barrs, Vanessa R.; van Doorn, Tineke M.; Houbraken, Jos; Kidd, Sarah E.; Martin, Patricia; Pinheiro, Maria Dolores; Richardson, Malcolm; Varga, Janos; Samson, Robert A.

    2013-01-01

    We describe a novel heterothallic species in Aspergillus section Fumigati, namely A. felis (neosartorya-morph) isolated from three host species with invasive aspergillosis including a human patient with chronic invasive pulmonary aspergillosis, domestic cats with invasive fungal rhinosinusitis and a

  4. First case report of isolated aspergillus dacryoadenitis

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    Ishan Acharya

    2016-01-01

    Full Text Available We report a case of isolated Aspergillus dacryoadenitis. A 23-year-old male presented with dull ache, diffuse swelling in superolateral quadrant of the right orbit and proptosis for 4 months. Ocular examination showed conjunctival congestion, discharge in the fornix and palpable lacrimal gland (LG mass. Routine hematological investigations followed by computed tomography scan of orbits were done. He did not respond to a course of systemic and topical antibiotics. Lateral orbitotomy with extended lid crease incision was performed with excision biopsy of LG. Abundant blackish material was found in the LG intraoperatively. The specimen was sent for histopathological examination (HPE. HPE report showed Aspergillus. Thorough ENT and systemic evaluation ruled out any other site with the fungus. To the best of our knowledge, this is the first case report of Aspergillus infection in LG.

  5. Three new species of Aspergillus section Flavi isolated from almonds and maize in Portugal

    Science.gov (United States)

    Three new aflatoxin-producing species belonging to Aspergillus section Flavi are described, Aspergillus mottae, Aspergillus sergii and Aspergillus transmontanensis. These species were isolated from Portuguese almonds and maize. An investigation examining morphology, extrolites and molecular data was...

  6. screening and improvement of local isolates of aspergillus niger

    African Journals Online (AJOL)

    DR. AMINU

    ABSTRACT. The study involved the screening of fourteen isolates of Aspergillus niger for citric acid production from glucose. The study was aimed at screening and improving local strains of Aspergillus niger with potential for citric acid production. All the isolates screened produced varying amounts of citric acid, the highest ...

  7. Aspergillus uvarum sp. nov., an uniseriate black Aspergillus species isolated from grapes in Europe

    DEFF Research Database (Denmark)

    Perrone, Giancarlo; Varga, János; Susca, Antonia

    2008-01-01

    A novel species, Aspergillus uvarum sp. nov., is described within Aspergillus section Nigri. This species can be distinguished from other black aspergilli based on internal transcribed spacers (ITS), beta-tubulin and calmodulin gene sequences, by AFLP analysis and by extrolite profiles. Aspergill...... atypical strains of Aspergillus aculeatus, CBS 114.80 and CBS 620.78, and was isolated from grape berries in Portugal, Italy, France, Israel, Greece and Spain. The type strain of Aspergillus uvarum sp. nov. is IMI 388523(T)=CBS 127591(T)= ITEM 4834(T)= IBT26606(T)....

  8. Aspergillus niger: an unusual cause of invasive pulmonary aspergillosis

    Science.gov (United States)

    Person, A. K.; Chudgar, S. M.; Norton, B. L.; Tong, B. C.; Stout, J. E.

    2010-01-01

    Infections due to Aspergillus species cause significant morbidity and mortality. Most are attributed to Aspergillus fumigatus, followed by Aspergillus flavus and Aspergillus terreus. Aspergillus niger is a mould that is rarely reported as a cause of pneumonia. A 72-year-old female with chronic obstructive pulmonary disease and temporal arteritis being treated with steroids long term presented with haemoptysis and pleuritic chest pain. Chest radiography revealed areas of heterogeneous consolidation with cavitation in the right upper lobe of the lung. Induced bacterial sputum cultures, and acid-fast smears and cultures were negative. Fungal sputum cultures grew A. niger. The patient clinically improved on a combination therapy of empiric antibacterials and voriconazole, followed by voriconazole monotherapy. After 4 weeks of voriconazole therapy, however, repeat chest computed tomography scanning showed a significant progression of the infection and near-complete necrosis of the right upper lobe of the lung. Serum voriconazole levels were low–normal (1.0 μg ml−1, normal range for the assay 0.5–6.0 μg ml−1). A. niger was again recovered from bronchoalveolar lavage specimens. A right upper lobectomy was performed, and lung tissue cultures grew A. niger. Furthermore, the lung histopathology showed acute and organizing pneumonia, fungal hyphae and oxalate crystallosis, confirming the diagnosis of invasive A. niger infection. A. niger, unlike A. fumigatus and A. flavus, is less commonly considered a cause of invasive aspergillosis (IA). The finding of calcium oxalate crystals in histopathology specimens is classic for A. niger infection and can be helpful in making a diagnosis even in the absence of conidia. Therapeutic drug monitoring may be useful in optimizing the treatment of IA given the wide variations in the oral bioavailability of voriconazole. PMID:20299503

  9. Cellulase Production by Aspergillus flavus Linn Isolate NSPR 101 ...

    African Journals Online (AJOL)

    Cellulase Production by Aspergillus flavus Linn Isolate NSPR 101 fermented in sawdust, bagasse and corncob. Tunde Victor Ojumu, Bamidele Ogbe Solomon, Eriola Betiku, Stephen Kolawole Layokun, Bamikole Amigun ...

  10. Aspergillus pragensis sp nov discovered during molecular reidentification of clinical isolates belonging to Aspergillus section Candidi

    DEFF Research Database (Denmark)

    Lyskova, Pavlina; Hubka, Vit; Kolarik, Miroslav

    2014-01-01

    The identity of nine clinical isolates recovered from Czech patients and presumptively identified as Aspergillus sp. section Candidi based on colony morphology was revised using sequences of beta-tubulin, calmodulin gene sequence, and internal transcribed spacer rDNA. Six isolates were from suspe...

  11. Detection of Polish clinical Aspergillus fumigatus isolates resistant to triazoles

    DEFF Research Database (Denmark)

    Nawrot, Urszula; Kurzyk, Ewelina; Arendrup, Maiken Cavling

    2018-01-01

    We studied the presence of triazole resistance of 121 Aspergillus fumigatus clinical isolates collected in two Polish cities, Warsaw and Wrocław, to determine if resistance is emerging in our country. We identified five itraconazole resistant isolates (4.13%) carrying the TR34/L98H alteration in ...

  12. Virulence Factors Detection inAspergillusIsolates from Clinical and Environmental Samples.

    Science.gov (United States)

    Raksha; Singh, Gurjeet; Urhekar, A D

    2017-07-01

    Pathogenesis of aspergillosis is dependent on various factors of the host (immune status) and virulence factors of the pathogen which could play a significant role in the pathogenesis of invasive aspergillosis. To study the virulence factors of Aspergillus species isolated from patient samples and environmental samples. This prospective and experimental study was carried out at Department of Microbiology, MGM Medical College and Hospital, Mumbai, Maharashtra, India, from July 2014 to June 2015. For detection of virulence factors of Aspergillus species, total 750 samples were included in this study (350 from patients and 400 samples from environment). Patient samples and hospital environment samples were subjected to standard methods for screening of Biofilm, Lipase, α-amylase, proteinase, haemolysin, phospholipase and pectinase. Statistical analysis was done using Chi-square test and SPSS (Version 17.0). American Type Culture Collection (ATCC) control of Aspergillus oryzae, Aspergillus niger and Aspergillus brasiliensis showed production of all virulence factors. In patient samples maximum virulence factor was produced i.e., α-amylase activity (89.74%) followed by proteinase activity (87.17%), biofilm production was (82.05%) haemolysin activity (79.48%), lipase activity (66.66%), pectinase activity and phospholipase activity (61.53%). In environment samples maximum virulence factor was produced i.e., proteinase activity (41.02%) followed by biofilm production was (38.46%), α-amylase activity (35.89%), haemolysin activity (33.33%), lipase activity (28.20%), phospholipase (25.64%) and pectinase activity (23.07%). The differences in patient and environment virulence factors were statistically significant (p-value Aspergillus species isolated from patient samples then environmental samples. This could be due to invasiveness nature of Aspergilli. Aspergillus niger was common isolates from both patient and environmental samples. Our study highlights the possible

  13. Virulence Factors Detection in Aspergillus Isolates from Clinical and Environmental Samples

    Science.gov (United States)

    Raksha; Urhekar, A.D.

    2017-01-01

    Introduction Pathogenesis of aspergillosis is dependent on various factors of the host (immune status) and virulence factors of the pathogen which could play a significant role in the pathogenesis of invasive aspergillosis. Aim To study the virulence factors of Aspergillus species isolated from patient samples and environmental samples. Materials and Methods This prospective and experimental study was carried out at Department of Microbiology, MGM Medical College and Hospital, Mumbai, Maharashtra, India, from July 2014 to June 2015. For detection of virulence factors of Aspergillus species, total 750 samples were included in this study (350 from patients and 400 samples from environment). Patient samples and hospital environment samples were subjected to standard methods for screening of Biofilm, Lipase, α–amylase, proteinase, haemolysin, phospholipase and pectinase. Statistical analysis was done using Chi-square test and SPSS (Version 17.0). Results American Type Culture Collection (ATCC) control of Aspergillus oryzae, Aspergillus niger and Aspergillus brasiliensis showed production of all virulence factors. In patient samples maximum virulence factor was produced i.e., α-amylase activity (89.74%) followed by proteinase activity (87.17%), biofilm production was (82.05%) haemolysin activity (79.48%), lipase activity (66.66%), pectinase activity and phospholipase activity (61.53%). In environment samples maximum virulence factor was produced i.e., proteinase activity (41.02%) followed by biofilm production was (38.46%), α-amylase activity (35.89%), haemolysin activity (33.33%), lipase activity (28.20%), phospholipase (25.64%) and pectinase activity (23.07%). The differences in patient and environment virulence factors were statistically significant (p-value Aspergillus species isolated from patient samples then environmental samples. This could be due to invasiveness nature of Aspergilli. Aspergillus niger was common isolates from both patient and environmental

  14. As a Rare Site of Invasive Fungal Infection, Chronic Granulomatous Aspergillus Synovitis: A Case Report

    Directory of Open Access Journals (Sweden)

    Aylin Canbolat Ayhan

    2013-06-01

    Full Text Available Aspergillus can causes invasive disease of various organs especially in patients with weakened immune systems. Aspergillus synovitis and arthritis are uncommon types of involvement due to this infection. Approches to fungal osteoarticular infections are based on only case reports. This paper presents a rare case of chronic granulomatous Aspergillus synovitis in an immunocompromised 5-year old girl who was treated for acute lymphoblastic leukemia.

  15. Invasive aspergillosis in the aortic arch with infectious Aspergillus lesions in pulmonary bullae

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    Isao Watanabe

    2015-03-01

    Full Text Available A patient with pulmonary bullae died of massive hemoptysis. At autopsy a hole was observed in the aortic wall. A microscopic examination indicated small Aspergillus lesions in pulmonary bullae and extensive necrotic lesions with Aspergillus hyphae in the media of the thoracic aorta. These findings led to a diagnosis of invasive aspergillosis in the aortic arch. This is a rare case in which Aspergillus invaded the aorta in a patient without hematologic neoplasms or neutropenia.

  16. Molecular analysis of Aspergillus section Flavi isolated from Brazil nuts.

    Science.gov (United States)

    Gonçalves, Juliana Soares; Ferracin, Lara Munique; Carneiro Vieira, Maria Lucia; Iamanaka, Beatriz Thie; Taniwaki, Marta Hiromi; Pelegrinelli Fungaro, Maria Helena

    2012-04-01

    Brazil nuts are an important export market in its main producing countries, including Brazil, Bolivia, and Peru. Approximately 30,000 tons of Brazil nuts are harvested each year. However, substantial nut contamination by Aspergillus section Flavi occurs with subsequent production of aflatoxins. In our study, Aspergillus section Flavi were isolated from Brazil nuts (Bertholletia excelsa), and identified by morphological and molecular means. We obtained 241 isolates from nut samples, 41% positive for aflatoxin production. Eighty-one isolates were selected for molecular investigation. Pairwise genetic distances among isolates and phylogenetic relationships were assessed. The following Aspergillus species were identified: A. flavus, A. caelatus, A. nomius, A. tamarii, A. bombycis, and A. arachidicola. Additionally, molecular profiles indicated a high level of nucleotide variation within β-tubulin and calmodulin gene sequences associated with high genetic divergence from RAPD data. Among the 81 isolates analyzed by molecular means, three of them were phylogenetically distinct from all other isolates representing the six species of section Flavi. A putative novel species was identified based on molecular profiles.

  17. Potential involvement of Aspergillus flavus laccases in peanut invasion at low water potential

    Science.gov (United States)

    Aspergillus flavus (Link) accumulates aflatoxins in peanuts, mainly affecting immature kernels during drought. Peanut invasion by A. flavus induces synthesis of phytoalexins, mostly stilbenoids, as a plant defense mechanism. Fungal laccases are often related to pathogenicity, and among other subst...

  18. Fatal invasive aspergillosis caused by Aspergillus niger after bilateral lung transplantation

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    Enora Atchade

    2017-09-01

    Full Text Available Aspergillus niger is usually considered to be a low virulence fungus, not commonly reported to cause invasive infections. Invasive pulmonary aspergillosis due to Aspergillus niger was diagnosed in a 43-year-old woman following bilateral lung transplantation. Intravenous voriconazole failed to control progression of the disease. Despite salvage therapy with a combination of voriconazole and caspofungin for 23 days, the patient developed massive hemoptysis leading to death. The authors report the clinical features and treatment of this case.

  19. Isolate-dependent growth, virulence, and cell wall composition in the human pathogen Aspergillus fumigatus.

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    Nansalmaa Amarsaikhan

    Full Text Available The ubiquitous fungal pathogen Aspergillus fumigatus is a mediator of allergic sensitization and invasive disease in susceptible individuals. The significant genetic and phenotypic variability between and among clinical and environmental isolates are important considerations in host-pathogen studies of A. fumigatus-mediated disease. We observed decreased radial growth, rate of germination, and ability to establish colony growth in a single environmental isolate of A. fumigatus, Af5517, when compared to other clinical and environmental isolates. Af5517 also exhibited increased hyphal diameter and cell wall β-glucan and chitin content, with chitin most significantly increased. Morbidity, mortality, lung fungal burden, and tissue pathology were decreased in neutropenic Af5517-infected mice when compared to the clinical isolate Af293. Our results support previous findings that suggest a correlation between in vitro growth rates and in vivo virulence, and we propose that changes in cell wall composition may contribute to this phenotype.

  20. MLST versus microsatellites for typing Aspergillus fumigatus isolates.

    Science.gov (United States)

    Klaassen, C H W

    2009-01-01

    In recent years, there has been a clear and growing tendency to use exact typing methods for discrimination between microbial isolates. Exact typing methods that yield an unambiguous typing result offer a number of advantages over conventional methods in the generation of typing data that is reproducible, portable and exchangeable. Two such methods are multi-locus sequence typing (MLST) and microsatellite-based typing. Here I will discuss the basic principles of both methods and compare them from a practical and performance point of view with respect to typing Aspergillus fumigatus isolates. Microsatellites offer the best available typing option by outperforming MLST in terms of speed, throughput, costs and discriminatory power. This latter advantage of microsatellites is a direct consequence of their inherent instability. This (in)stability of individual microsatellite markers and alleles should be taken into account in the interpretation of microsatellite-based typing data.

  1. Evaluation of the detection techniques of toxigenic Aspergillus isolates

    African Journals Online (AJOL)

    However, our results showed that use of these techniques were not sufficiently sensitive for all Aspergillus species and suggested a simple thin layer chromatography (TLC) as a sensitive and reliable technique for detection of aflatoxins and OTA produced by Aspergillus and Eurotium species. Key words: Aspergillus ...

  2. Polyphasic approach to the identification of Aspergillus section Flavi isolated from Brazil nuts.

    Science.gov (United States)

    Baquião, Arianne Costa; de Oliveira, Maitê Martins Melo; Reis, Tatiana Alves; Zorzete, Patricia; Diniz Atayde, Danielle; Correa, Benedito

    2013-08-15

    The aim of this study was to use a polyphasic approach to identify Aspergillus section Flavi isolated from Brazil nuts collected in the Amazon forest: investigation of macro- and microscopic morphology, production of extrolites, heat-resistance fungi, and sequencing of DNA regions. The following Aspergillus section Flavi species were identified: Aspergillus flavus (75.5%), Aspergillus nomius (22.3%), and Aspergillus parasiticus (2.2%). All A. nomius and A. parasiticus isolates produced aflatoxins B and G, but not cyclopiazonic acid (CPA). A. flavus isolates were more diversified and a high frequency of mycotoxigenic strains was observed. The polyphasic approach permitted the reliable identification of section Flavi species. The rate of mycotoxigenic strains was high (92.7%) and mainly included A. flavus strains producing elevated levels of aflatoxins and CPA. These results highlight the possibility of co-occurrence of both toxins, increasing their potential toxic effect in this commodity. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. Immunoproteome of Aspergillus fumigatus Using Sera of Patients with Invasive Aspergillosis

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    Emylli D. Virginio

    2014-08-01

    Full Text Available Invasive aspergillosis is a life-threatening lung or systemic infection caused by the opportunistic mold Aspergillus fumigatus. The disease affects mainly immunocompromised hosts, and patients with hematological malignances or who have been submitted to stem cell transplantation are at high risk. Despite the current use of Platelia™ Aspergillus as a diagnostic test, the early diagnosis of invasive aspergillosis remains a major challenge in improving the prognosis of the disease. In this study, we used an immunoproteomic approach to identify proteins that could be putative candidates for the early diagnosis of invasive aspergillosis. Antigenic proteins expressed in the first steps of A. fumigatus germination occurring in a human host were revealed using 2-D Western immunoblots with the serum of patients who had previously been classified as probable and proven for invasive aspergillosis. Forty antigenic proteins were identified using mass spectrometry (MS/MS. A BLAST analysis revealed that two of these proteins showed low homology with proteins of either the human host or etiological agents of other invasive fungal infections. To our knowledge, this is the first report describing specific antigenic proteins of A. fumigatus germlings that are recognized by sera of patients with confirmed invasive aspergillosis who were from two separate hospital units.

  4. Aspergillus pragensis sp nov discovered during molecular reidentification of clinical isolates belonging to Aspergillus section Candidi

    Czech Academy of Sciences Publication Activity Database

    Hubka, Vít; Lysková, P.; Frisvad, J.C.; Peterson, S.W.; Skořepová, M.; Kolařík, Miroslav

    2014-01-01

    Roč. 52, č. 6 (2014), s. 565-576 ISSN 1369-3786 R&D Projects: GA MŠk(CZ) EE2.3.20.0055; GA MŠk(CZ) EE2.3.30.0003 Institutional support: RVO:61388971 Keywords : Aspergillus candidus * Aspergillus tritici * antifungal susceptibility testing Subject RIV: EE - Microbiology, Virology Impact factor: 2.335, year: 2014

  5. Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates.

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    Dong Ying Wang

    Full Text Available Aspergillus flavus is second only to A. fumigatus in causing invasive aspergillosis and it is the major agent responsible for fungal sinusitis, keratitis and endophthalmitis in many countries in the Middle East, Africa and Southeast Asia. Despite the growing challenge due to A. flavus, data on the molecular epidemiology of this fungus remain scarce. The objective of the present study was to develop a new typing method based on the detection of VNTR (Variable number tandem repeat markers. Eight VNTR markers located on 6 different chromosomes (1, 2, 3, 5, 7 and 8 of A. flavus were selected, combined by pairs for multiplex amplifications and tested on 30 unrelated isolates and six reference strains. The Simpson index for individual markers ranged from 0.398 to 0.818. A combined loci index calculated with all the markers yielded an index of 0.998. The MLVA (Multiple Locus VNTR Analysis technique proved to be specific and reproducible. In a second time, a total of 55 isolates from Chinese avian farms and from a Tunisian hospital have been evaluated. One major cluster of genotypes could be defined by using the graphing algorithm termed Minimum Spanning Tree. This cluster comprised most of the isolates collected in an avian farm in southern China. The MLVA technique should be considered as an excellent and cost-effective typing method that could be used in many laboratories without the need for sophisticated equipment.

  6. Invasive Aspergillus terreus morphological transitions and immunoadaptations mediating antifungal resistance

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    Bengyella L

    2017-11-01

    Full Text Available Louis Bengyella,1–3 Elsie Laban Yekwa,4 Muhammad Nasir Subhani,5 Ernest Tambo,6,7 Kiran Nawaz,5 Bakoena Ashton Hetsa,2 Sehrish Iftikhar,5 Sayanika Devi Waikhom,1 Pranab Roy8 1Department of Biomedical Science, The School of Basic and Biomedical Sciences, University of Health and Allied Sciences, Ho, Ghana; 2Department of Biotechnology, Faculty of Applied and Computer Sciences, Vaal University of Technology, Vanderbijlpark, South Africa; 3Department of Biotechnology, University of Burdwan, Bardhaman, India; 4Division of Medical Virology, Stellenbosch University, Stellenbosch, South Africa; 5Department of Plant Pathology, Institute of Agricultural Sciences, University of the Punjab, Lahore, Pakistan; 6Department of Biochemistry and Pharmaceutical Sciences, Université des Montagnes, Bangangté, 7Department of Communications, Africa Disease Intelligence and Surveillance, Communication and Response Institute, Yaoundé, Cameroon; 8Department of Biotechnology, Haldia Institute of Technology, Haldia, India Background and aims: Aspergillus terreus Thom is a pathogen of public health and agricultural importance for its seamless abilities to expand its ecological niche. The aim of this study was holistically to investigate A. terreus morphological and immunoadaptations and their implication in antifungal resistance and proliferation during infection.Materials and methods: In-depth unstructured mining of relevant peer-reviewed literature was performed for A. terreus morphological, immune, resistance, and genetic diversity based on the sequenced calmodulin-like gene.Results: Accessory conidia and phialidic conidia produced by A. terreus confer discrete antifungal resistance that ensures survivability during therapies. Interestingly, by producing unique metabolites such as Asp–melanin and terretonin, A. terreus is capable of hijacking macrophages and scavenging iron, respectively. As such, A. terreus has established a rare mechanism to mitigate

  7. Challenges in microbiological diagnosis of invasive Aspergillus infections [version 1; referees: 2 approved

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    Alexandre Alanio

    2017-02-01

    Full Text Available Invasive aspergillosis (IA has been increasingly reported in populations other than the historical hematology patients and there are new questions about the performance of microbiological tools. Microscopy and culture have been completed by biomarkers, either antigens or DNA, and in blood or respiratory specimens or both. First studied in hematology, the antigen galactomannan performance in serum is low in other patient populations where the pathophysiology of the infection can be different and the prevalence of IA is much lower. DNA detection with polymerase chain reaction (PCR in blood or serum (or both has reached a certain level of acceptance thanks to consensus methods based on real-time quantitative PCR (qPCR. When used on respiratory specimens, galactomannan and qPCR depend on standardization of the sampling and the diverse mycological procedures. Thus, culture remains the main diagnostic criterion in critically ill patients. The current trend toward more effective anti-mold prophylaxis in hematology hampers the yield of a screening strategy, as is usually performed in hematology. Therefore, circulating biomarkers as confirmatory tests should be considered and their performance should be reappraised in each new setting. The use of azole prophylaxis also raises the issue of selecting azole-resistance Aspergillus fumigatus isolates. Ideally, the biomarkers will be more efficient when individual genetic risks of IA are defined. Culture, though not standardized, remains a key element for the diagnosis of IA and has the advantage to easily detect molds other than A. fumigatus. It is still unclear whether next-generation sequencing will replace culture in the future.

  8. Morphological and molecular identification of filamentous Aspergillus flavus and Aspergillus parasiticus isolated from compound feeds in South Africa.

    Science.gov (United States)

    Iheanacho, Henry E; Njobeh, Patrick B; Dutton, Francis M; Steenkamp, Paul A; Steenkamp, Lucia; Mthombeni, Julian Q; Daru, Barnabas H; Makun, Anthony H

    2014-12-01

    Isolation of filamentous species of two Aspergillum genera from compound feeds produced in South Africa, and subsequent extraction of their individual DNA in this study, presents a simple but rapid molecular procedure for high through-put analysis of the individual morphological forms. DNA was successfully isolated from the Aspergillus spp. from agar cultures by use of a commercial kit. Agarose gel electrophoresis fractionation of the fungi DNA, showed distinct bands. The DNA extracted by this procedure appears to be relatively pure with a ratio absorbance at 260 and 280 nm. However, the overall morphological and molecular data indicated that 67.5 and 51.1% of feed samples were found to be contaminated with Aspergillus flavus and Aspergillus parasiticus, respectively, with poultry feed having the highest contamination mean level of 5.7 × 105 CFU/g when compared to cattle (mean: 4.0 × 106 CFU/g), pig (mean: 2.7 × 104 CFU/g) and horse (1.0 × 102 CFU) feed. This technique presents a readily achievable, easy to use method in the extraction of filamentous fungal DNA and it's identification. Hence serves as an important tool towards molecular study of these organisms for routine analysis check in monitoring and improving compound feed quality against fungal contamination. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Isolation and properties of pectinases from the fungus Aspergillus japonicus.

    Science.gov (United States)

    Semenova, M V; Grishutin, S G; Gusakov, A V; Okunev, O N; Sinitsyn, A P

    2003-05-01

    Using anion-exchange chromatography on different carriers and phenyl-Sepharose hydrophobic chromatography, five pectolytic enzymes were isolated from the culture liquid of a mutant strain of Aspergillus japonicus: two endo-polygalacturonases (I and II, 38 and 65 kD, pI 5.6 and 3.3), pectin lyase (50 kD, pI 3.8), and two pectinesterases (I and II) with similar molecular weights (46 and 47 kD) and the same pI (3.8). The pectinesterases apparently represent two isoforms of the same enzyme. All purified enzymes were homogenous according to SDS-PAGE and polyacrylamide gel-IEF, except for endo-polygalacturonase II that gave two bands on isoelectric focusing, but one band on electrophoresis. All enzymes had maximal activity in an acid medium (at pH 4.0-5.5). The pectin lyase and pectinesterase were stable at 40-50 degrees C. The thermal stability of both endo-polygalacturonases was much lower (after 3 h of incubation at 30 degrees C, endo-polygalacturonases I and II lost 40 and 10% of the activity, respectively). The activity of endo-polygalacturonases I and II towards polygalacturonic acid strongly depended on NaCl concentration (optimal concentration of the salt was 0.1-0.2 M); the enzymes were also capable of reducing the viscosity of pectin solution, but rather slowly. The pectin lyase had no activity towards polygalacturonic acid. The activity of the pectin lyase increased with increasing degree of methylation of pectins. Both endo-polygalacturonases demonstrated synergism with the pectinesterase during the hydrolysis of highly methylated pectin. On the contrary, in the mixture of pectin lyase and pectinesterase an antagonism between the two enzymes was observed.

  10. Atypical Aspergillus parasiticus isolates from pistachio with aflR gene nucleotide insertion identical to Aspergillus sojae

    Science.gov (United States)

    Aflatoxins are the most toxic and carcinogenic secondary metabolites produced primarily by the filamentous fungi Aspergillus flavus and Aspergillus parasiticus. The toxins cause devastating economic losses because of strict regulations on distribution of contaminated products. Aspergillus sojae are...

  11. Aspergillus fumigatus Does Not Require Fatty Acid Metabolism via Isocitrate Lyase for Development of Invasive Aspergillosis▿

    Science.gov (United States)

    Schöbel, Felicitas; Ibrahim-Granet, Oumaïma; Avé, Patrick; Latgé, Jean-Paul; Brakhage, Axel A.; Brock, Matthias

    2007-01-01

    Aspergillus fumigatus is the most prevalent airborne filamentous fungus causing invasive aspergillosis in immunocompromised individuals. Only a limited number of determinants directly associated with virulence are known, and the metabolic requirements of the fungus to grow inside a host have not yet been investigated. Previous studies on pathogenic microorganisms, i.e., the bacterium Mycobacterium tuberculosis and the yeast Candida albicans, have revealed an essential role for isocitrate lyase in pathogenicity. In this study, we generated an isocitrate lyase deletion strain to test whether this strain shows attenuation in virulence. Results have revealed that isocitrate lyase from A. fumigatus is not required for the development of invasive aspergillosis. In a murine model of invasive aspergillosis, the wild-type strain, an isocitrate lyase deletion strain, and a complemented mutant strain were similarly effective in killing mice. Moreover, thin sections demonstrated invasive growth of all strains. Additionally, thin sections of lung tissue from patients with invasive aspergillosis stained with anti-isocitrate lyase antibodies remained negative. From these results, we cannot exclude the use of lipids or fatty acids as a carbon source for A. fumigatus during invasive growth. Nevertheless, test results do imply that the glyoxylate cycle from A. fumigatus is not required for the anaplerotic synthesis of oxaloacetate under infectious conditions. Therefore, an antifungal drug inhibiting fungal isocitrate lyases, postulated to act against Candida infections, is assumed to be ineffective against A. fumigatus. PMID:17178786

  12. Removal of trace element by isolates of Aspergillus brasiliensis ...

    African Journals Online (AJOL)

    Coffee beans processing generates a large volume of wastewater composed of trace elements which can be detrimental to human health. The present study aimed at evaluating the capacity of strains of Aspergillus brasiliensis and Penicillium citrinum in tolerating and removing trace elements namely: Cu, Mn and Zn from ...

  13. Prevalence of potential toxigenic Aspergillus species isolated from ...

    African Journals Online (AJOL)

    ADEYEYE

    2015-12-17

    Dec 17, 2015 ... *Correspondence: Tel.: +234 8032539564, E-mail: rabiuabdul2005@yahoo.com. Abstract. The objective of this study was to investigate the prevalence and risk factors of fungal organisms (especially. Aspergillus species) in feeds used in poultry farms in Sokoto metropolis. During a period of 12 months ( ...

  14. Glucoamylase production by a newly isolated strain of Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Sinkar, V.P.; Lewis, N.F.

    1982-01-01

    Glucoamylase production by Aspergillus niger 57 was studied in complex and synthetic media under stationary vs. submerged conditions. Stationary cultivation resulted in significantly greater yields than did submerged culture. Crude enzyme activity was optimum at 60 degrees and pH 4.0.

  15. Aflatoxin B1 producing potential of Aspergillus flavus strains isolated ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-07-20

    Jul 20, 2009 ... However, 65% of the strains produced AFB1 on Czapek's agar, 53% of the strains on potato dextrose agar (PDA) and none of the strains on Aspergillus flavus and parasiticus agar media (AFPA). The strain, DRAf 009 produced maximum AFB1 (4.0 – 40 µg/g agar) on all the agar media tested. Five strains of ...

  16. Aspergillus species isolated from mangrove forests in Borneo Island, Sarawak, Malaysia

    Directory of Open Access Journals (Sweden)

    J.S.S. Seelan

    2009-06-01

    Full Text Available A study on the occurrence of Aspergillus spp. on selected mangrove forests in Sarawak was conducted to find out their diversity and distribution. Samples were obtained from mangrove soils and leaf litters at different locations, i.e. Sematan, Lundu, Kampung Bako, Bako in Sarawak. Soil and leaf litter samples were taken randomly at different locations with five replicates from each area. A total of 138 isolates of Aspergillus species were obtained from the soil and leaf litter samples by using direct plating and Warcup method. Based on both macroscopic and microscopic observations, using an identification key, individual isolates were classified within the genus Aspergillus, belonging to three subgenera, four sections and five species. The fungi isolates were identified as A. terreus, A. flavipes, A. carneus, A. fumigatus and A. clavatus. The most frequent isolated species was A. flavipes (63.04%, followed by A. fumigatus (16.7%, A. terreus (13.04%, A. carneus (5.8% and A. clavatus (1.44%. All of the isolated Aspergillus species grew well on MEA and CYA at 25°C. A. carneus produced reddish sclerotia on MEA after seven days and this could be used as an important characteristic in this species identification. A. clavatus from mangrove soil in Kampung Bako has shown long conidiophores (ranging from 3-5 cm with swollen hyphal structures, while A. clavatus from Sematan area has shorter conidiophores (ranging from 2.5-3.5 cm on MEA.

  17. Induction, isolation, and characterization of aspergillus niger mutant strains producing elevated levels of beta-galactosidase.

    OpenAIRE

    Nevalainen, K M

    1981-01-01

    An Aspergillus niger mutant strain, VTT-D-80144, with an improvement of three- to fourfold in the production of extracellular beta-galactosidase was isolated after mutagenesis. The production of beta-galactosidase by this mutant was unaffected by fermentor size, and the enzyme was also suitable for immobilization.

  18. The isolation and improvement of Aspergillus niger by radiation for higher production of citric acid

    International Nuclear Information System (INIS)

    Radziah A; Foziah Ali; Zainab H

    2000-01-01

    Local citric acid producer of fungal strain Aspergillus niger have been successfully isolated from stale bread and onion. The isolates, designated as SB 1 and NN I showed a potential performance for citric acid production of 49% and 52% yield respectively, in shake flask studies. The strain improvement on NN1 was carried out by radiation induced mutation by gamma rays at LD 5 0 of 1.28 kGy

  19. Karnatakafuran A and B: Two novel dibenzofurans isolated from the fungus Aspergillus karnatakaensis

    DEFF Research Database (Denmark)

    Manniche, S.; Sprogøe, K.; Dalsgaard, Petur

    2004-01-01

    Karnatakafurans A (1) and B (2), two novel dibenzofurans, have been isolated from the Specie Novum Aspergillus karnatakaensis Frisvad. The compounds were the major secondary metabolites and were isolated through UV-guided fractionation. of the organic extract. The structures were elucidated by sp...... by spectroscopic methods including MS and NMR. The compounds were tested for antimicrobial and antimalarial activity and proved to be moderately active against Plasmodium falciparum....

  20. [Mixed invasive fungal infection due to Rhizomucor pusillus and Aspergillus niger in an immunocompetent patient].

    Science.gov (United States)

    Pozo-Laderas, Juan Carlos; Pontes-Moreno, Antonio; Robles-Arista, Juan Carlos; Bautista-Rodriguez, M Dolores; Candau-Alvarez, Alberto; Caro-Cuenca, Maria Teresa; Linares-Sicilia, María José

    2015-01-01

    Mucormycosis infections are rare in immunocompetent patients, and very few cases of mucormycosis associated with aspergillosis in non-haematological patients have been reported. A 17-year-old male, immunocompetent and without any previously known risk factors, was admitted to hospital due to a seizure episode 11 days after a motorcycle accident. He had a complicated clinical course as he had a mixed invasive fungal infection with pulmonary involvement due to Aspergillus niger and disseminated mucormycosis due to Rhizomucor pusillus (histopathological and microbiological diagnosis in several non-contiguous sites). He was treated with liposomal amphotericin B for 7 weeks (total cumulative dose >10 g) and required several surgical operations. The patient survived and was discharged from ICU after 5 months and multiple complications. Treatment with liposomal amphotericin B and aggressive surgical management achieved the eradication of a mixed invasive fungal infection. However, we emphasise the need to maintain a higher level of clinical suspicion and to perform microbiological techniques for early diagnosis of invasive fungal infections in non-immunocompromised patients, in order to prevent spread of the disease and the poor prognosis associated with it. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  1. Invasion of Solanum tuberosum L. by Aspergillus terreus: a microscopic and proteomics insight on pathogenicity.

    Science.gov (United States)

    Louis, Bengyella; Waikhom, Sayanika Devi; Roy, Pranab; Bhardwaj, Pardeep Kumar; Singh, Mohendro Wakambam; Chandradev, Sharma K; Talukdar, Narayan Chandra

    2014-06-10

    Aspergillus terreus is one of the most harmful filamentous fungal pathogen of humans, animals and plants. Recently, researchers have discovered that A. terreus can cause foliar blight disease in potato (Solanum tuberosum L.). We used light and scanning electron microscopy, and performed proteomics analysis in an attempt to dissect the invasion process of A. terreus in this important crop. Microscopic study revealed that invasion of leaf tissue is marked by rapid germination of A. terreus phialidic conidia (PC) by 4 h after inoculation. By 8 h after inoculation, primary germ tubes from PC differentiated into irregular protuberance, often displayed stomata atropism, and failed to penetrate via the epidermal cells. Colonization of leaf tissues was associated with high rate of production of accessory conidia (AC). These analyses showed the occurrence of a unique opposing pattern of AC, tissue-specific and produced on melanized colonizing hyphae during the infection of leaf tissue. A significant proteome change hallmarked by differential expression of class I patatin, lipoxygenase, catalase-peroxidase complex, and cysteine proteinase inhibitor were observed during tuber colonization. These proteins are often involved in signal transduction pathways and crosstalk in pathogenic responses. A. terreus abundantly produced AC and multipolar germinating PC to invade potato leaf tissue. Additionally, A. terreus differentially induced enzymes in potato tuber during colonization which facilitates rapid disease development.

  2. Molecular identification and amphotericin B susceptibility testing of clinical isolates of Aspergillus from 11 hospitals in Korea.

    Science.gov (United States)

    Heo, Min Seok; Shin, Jong Hee; Choi, Min Ji; Park, Yeon Joon; Lee, Hye Soo; Koo, Sun Hoe; Lee, Won Gil; Kim, Soo Hyun; Shin, Myung Geun; Suh, Soon Pal; Ryang, Dong Wook

    2015-11-01

    We investigated the species distribution and amphotericin B (AMB) susceptibility of Korean clinical Aspergillus isolates by using two Etests and the CLSI broth microdilution method. A total of 136 Aspergillus isolates obtained from 11 university hospitals were identified by sequencing the internal transcribed spacer (ITS) and β-tubulin genomic regions. Minimal inhibitory concentrations (MICs) of AMB were determined in Etests using Mueller-Hinton agar (Etest-MH) and RPMI agar (Etest-RPG), and categorical agreement with the CLSI method was assessed by using epidemiological cutoff values. ITS sequencing identified the following six Aspergillus species complexes: Aspergillus fumigatus (42.6% of the isolates), A. niger (23.5%), A. flavus (17.6%), A. terreus (11.0%), A. versicolor (4.4%), and A. ustus (0.7%). Cryptic species identifiable by β-tubulin sequencing accounted for 25.7% (35/136) of the isolates. Of all 136 isolates, 36 (26.5%) had AMB MICs of ≥2 μg/mL by the CLSI method. The categorical agreement of Etest-RPG with the CLSI method was 98% for the A. fumigatus, A. niger, and A. versicolor complexes, 87% for the A. terreus complex, and 37.5% for the A. flavus complex. That of Etest-MH was ≤75% for the A. niger, A. flavus, A. terreus, and A. versicolor complexes but was higher for the A. fumigatus complex (98.3%). Aspergillus species other than A. fumigatus constitute about 60% of clinical Aspergillus isolates, and reduced AMB susceptibility is common among clinical isolates of Aspergillus in Korea. Molecular identification and AMB susceptibility testing by Etest-RPG may be useful for characterizing Aspergillus isolates of clinical relevance.

  3. Real-time PCR-based method for rapid detection of Aspergillus niger and Aspergillus welwitschiae isolated from coffee.

    Science.gov (United States)

    von Hertwig, Aline Morgan; Sant'Ana, Anderson S; Sartori, Daniele; da Silva, Josué José; Nascimento, Maristela S; Iamanaka, Beatriz Thie; Pelegrinelli Fungaro, Maria Helena; Taniwaki, Marta Hiromi

    2018-03-23

    Some species from Aspergillus section Nigri are morphologically very similar and altogether have been called A. niger aggregate. Although the species included in this group are morphologically very similar, they differ in their ability to produce mycotoxins and other metabolites and their taxonomical status has evolved continuously. Among them, A. niger and A. welwitschiae are ochratoxin A and fumonisin B 2 producers and their detection and/or identification is of crucial importance for food safety. The aim of this study was the development of a real-time PCR-based method for simultaneous discrimination of A. niger and A. welwitschiae from other species of the A. niger aggregate isolated from coffee beans. One primer pair and a hybridization probe specific for detection of A. niger and A. welwitschiae strains were designed based on the BenA gene sequences, and used in a Real-time PCR assay for the rapid discrimination between both these species from all others of the A. niger aggregate. The Real-time PCR assay was shown to be 100% efficient in discriminating the 73 isolates of A. niger/A. welwitschiae from the other A. niger aggregate species analyzed as a negative control. This result testifies to the use of this technique as a good tool in the rapid detection of these important toxigenic species. Copyright © 2018 Elsevier B.V. All rights reserved.

  4. Diagnosis of invasive aspergillus tracheobronchitis facilitated by endobronchial ultrasound-guided transbronchial needle aspiration: a case report

    Directory of Open Access Journals (Sweden)

    Casal Roberto F

    2009-11-01

    Full Text Available Abstract Introduction Invasive pulmonary aspergillosis is the most common form of infection by Aspergillus species among immunocompromised patients. Although this infection frequently involves the lung parenchyma, it is unusual to find it limited to the tracheobronchial tree, a condition known as invasive aspergillus tracheobronchitis. Case presentation A 65 year-old Hispanic man from Bolivia with a history of chronic lymphocytic leukemia developed cough and malaise eight months after having an allogenic stem cell transplant. A computed tomography of the chest revealed an area of diffuse soft tissue thickening around the left main stem bronchus, which was intensely fluorodeoxyglucose-avid on positron emission tomography scanning. An initial bronchoscopic exam revealed circumferential narrowing of the entire left main stem bronchus with necrotic and friable material on the medial wall. Neither aspirates from this necrotic area nor bronchial washing were diagnostic. A second bronchoscopy with endobronchial ultrasound evidenced a soft tissue thickening on the medial aspect of the left main stem bronchus underlying the area of necrosis visible endoluminally. Endobronchial ultrasound-guided transbronchial needle aspiration performed in this area revealed multiple fungal elements suggestive of Aspergillus species. Conclusion We describe the first case of invasive aspergillus tracheobronchitis in which the diagnosis was facilitated by the use of endobronchial ultrasound guided trans-bronchial needle aspiration. To the best of our knowledge, we are also presenting the first positron emission tomography scan images of this condition in the literature. We cautiously suggest that endobronchial ultrasound imaging may be a useful tool to evaluate the degree of invasion and the involvement of vascular structures in these patients prior to bronchoscopic manipulation of the affected areas in an effort to avoid potentially fatal hemorrhage.

  5. Invasive Aspergillus flavus sinusitis: case report in a patient with biphenotypic acute leukemia Sinusite invasiva por Aspergillus flavus: relato de um caso associado a leucemia aguda bifenotípica

    Directory of Open Access Journals (Sweden)

    Melissa Orzechowski Xavier

    2009-02-01

    Full Text Available Here we report a case of invasive pansinusitis with proptosis of the right eye caused by Aspergillus flavus in an immunocompromised patient with acute biphenotypic leukemia without aggressive therapy response.Descreve-se um caso de pansinusite invasiva com proptose do globo ocular direito causado por Aspergillus flavus em um paciente imunossuprimido com leucemia aguda bifenotípica sem resposta a terapia agressiva.

  6. Genetic isolation among sympatric vegetative compatibility groups of the aflatoxin-producing fungus Aspergillus flavus.

    Science.gov (United States)

    Grubisha, L C; Cotty, P J

    2010-01-01

    Aspergillus flavus, a fungal pathogen of animals and both wild and economically important plants, is most recognized for producing aflatoxin, a cancer-causing secondary metabolite that contaminates food and animal feed globally. Aspergillus flavus has two self/nonself recognition systems, a sexual compatibility system and a vegetative incompatibility system, and both play a role in directing gene flow in populations. Aspergillus flavus reproduces clonally in wild and agricultural settings, but whether a cryptic sexual stage exists in nature is currently unknown. We investigated the distribution of genetic variation in 243 samples collected over 4 years from three common vegetative compatibility groups (VCGs) in Arizona and Texas from cotton using 24 microsatellite loci and the mating type locus (MAT) to assess population structure and potential gene flow among A. flavus VCGs in sympatric populations. All isolates within a VCG had the same mating type with OD02 having MAT1-2 and both CG136 and MR17 having MAT1-1. Our results support the hypothesis that these three A. flavus VCGs are genetically isolated. We found high levels of genetic differentiation and no evidence of gene flow between VCGs, including VCGs of opposite mating-type. Our results suggest that these VCGs diverged before domestication of agricultural hosts (>10,000 yr bp).

  7. Screening and Improvement of Local Isolates of Aspergillus Niger ...

    African Journals Online (AJOL)

    The isolate CP3 was chosen for further studies. Strain improvement studies which was based on exposure of CP3 to ultraviolet irradiation for 10, 30 and 45 minutes increased citric acid yield to 22.20g/l (48.89% increase) by mutant M45 followed by a citric acid yield of 16.22g/l by the 30 minutes mutant (M30) and the least ...

  8. Methionine and glucose transport by isolated intestinal brush border membrane vesicles from pigs and lambs fed an Aspergillus product

    OpenAIRE

    Jang, Insurk

    1993-01-01

    This study was designed to determine whether feeding an Aspergillus product would influence growth or feed utilization and intestinal mucosal cell function as indicated by uptake of methionine and glucose by isolated intestinal brush border membrane vesicles (BBMV). In Experiment 1, 24 weanling pigs were paired by sex, BW, and litter and were allotted, within pairs, to either an 18% CP corn-soy diet (control) or the same diet supplemented (.15%) with an Aspergillus product. There were no diff...

  9. Inhibitory activity of compounds isolated from Polymnia sonchifolia on aflatoxin production by Aspergillus flavus

    Directory of Open Access Journals (Sweden)

    Pak Adriana

    2006-01-01

    Full Text Available Polymnia sonchifolia, commonly known as ";yacon";, was originally cultivated at Andes moutains in South America. Recently, the specie attracted worldwide attention because of its wide range of uses, for example in the control of diabetes melitus, besides the antifungal and pesticidal compounds were found in the leaves. This study describes the identification of two flavonoids: 3', 5, 7 trihydroxy-3, 4'-dimethoxyflavone (compound 1 and 3', 4', 5- trihydroxy-7-methoxy flavanone (compound 2 and two sesquiterpenes lactones: enhydrin (compound 3 and a mixture of enhydrin and uvedalin (compound 4 isolated from Polymnia sonchifolia leaves and their effects on the aflatoxin production by Aspergillus flavus. The identification of the compounds were achieved by ¹H and 13C NMR. All compounds were tested in different concentration, to evaluate the growth of Aspergillus flavus culture and the production of aflatoxin. The compound 1, at the concentration 15 mug/mL, inhibited 25% of the aflatoxin B1 production (p<0.01. The compound 4 inhibited 34% and 76% of the fungal growth and AFB1 production respectively. These results show that Polymnia sonchifolia can be used for the development of agents to control aflatoxin B1 production by Aspergillus flavus.

  10. Abcès épidural compressif isolé à aspergillus | N'da | Revue ...

    African Journals Online (AJOL)

    Spinal epidural abscess by Aspergillus is a rare disease on immunocompetent individuals. The diagnosis remains difficult. In immunocompromised patient, this affection occurs in invasive aspergillosis. The authors report a single case of retro vertebral aspergillar abscess revealed by a thoracic spinal compression in a 55 ...

  11. Notable fibrolytic enzyme production by Aspergillus spp. isolates from the gastrointestinal tract of beef cattle fed in lignified pastures

    Science.gov (United States)

    Abrão, Flávia Oliveira; Pessoa, Moisés Sena; dos Santos, Vera Lúcia; de Freitas Júnior, Luiz Fernando; Barros, Katharina de Oliveira; Hughes, Alice Ferreira da Silva; Silva, Thiago Dias; Rodriguez, Norberto Mário

    2017-01-01

    Fungi have the ability to degrade vegetal cell wall carbohydrates, and their presence in the digestive tract of ruminants can minimize the effects of lignified forage on ruminal fermentation. Here, we evaluated enzyme production by Aspergillus spp. isolates from the digestive tracts of cattle grazed in tropical pastures during the dry season. Filamentous fungi were isolated from rumen and feces by culture in cellulose-based medium. Ninety fungal strains were isolated and identified by rDNA sequence analysis, microculture, or both. Aspergillus terreus was the most frequently isolated species, followed by Aspergillus fumigatus. The isolates were characterized with respect to their cellulolytic, xylanolytic, and lignolytic activity through qualitative evaluation in culture medium containing a specific corresponding carbon source. Carboxymethyl cellulase (CMCase) activity was quantified by the reducing sugar method. In the avicel and xilan degradation test, the enzyme activity (EA) at 48 h was significantly higher other periods (P < 0.05). Intra- and inter-specific differences in EA were verified, and high levels of phenoloxidases, which are crucial for lignin degradation, were observed in 28.9% of the isolates. Aspergillus terreus showed significantly higher EA for avicelase (3.96 ±1.77) and xylanase (3.13 ±.091) than the other Aspergillus species at 48 h of incubation. Isolates AT13 and AF69 showed the highest CMCase specific activity (54.84 and 33.03 U mg-1 protein, respectively). Selected Aspergillus spp. isolates produced remarkable levels of enzymes involved in vegetal cell wall degradation, suggesting their potential as antimicrobial additives or probiotics in ruminant diets. PMID:28850605

  12. Notable fibrolytic enzyme production by Aspergillus spp. isolates from the gastrointestinal tract of beef cattle fed in lignified pastures.

    Directory of Open Access Journals (Sweden)

    Flávia Oliveira Abrão

    Full Text Available Fungi have the ability to degrade vegetal cell wall carbohydrates, and their presence in the digestive tract of ruminants can minimize the effects of lignified forage on ruminal fermentation. Here, we evaluated enzyme production by Aspergillus spp. isolates from the digestive tracts of cattle grazed in tropical pastures during the dry season. Filamentous fungi were isolated from rumen and feces by culture in cellulose-based medium. Ninety fungal strains were isolated and identified by rDNA sequence analysis, microculture, or both. Aspergillus terreus was the most frequently isolated species, followed by Aspergillus fumigatus. The isolates were characterized with respect to their cellulolytic, xylanolytic, and lignolytic activity through qualitative evaluation in culture medium containing a specific corresponding carbon source. Carboxymethyl cellulase (CMCase activity was quantified by the reducing sugar method. In the avicel and xilan degradation test, the enzyme activity (EA at 48 h was significantly higher other periods (P < 0.05. Intra- and inter-specific differences in EA were verified, and high levels of phenoloxidases, which are crucial for lignin degradation, were observed in 28.9% of the isolates. Aspergillus terreus showed significantly higher EA for avicelase (3.96 ±1.77 and xylanase (3.13 ±.091 than the other Aspergillus species at 48 h of incubation. Isolates AT13 and AF69 showed the highest CMCase specific activity (54.84 and 33.03 U mg-1 protein, respectively. Selected Aspergillus spp. isolates produced remarkable levels of enzymes involved in vegetal cell wall degradation, suggesting their potential as antimicrobial additives or probiotics in ruminant diets.

  13. Molecular identification of Aspergillus spp. isolated from coffee beans Identificação molecular de Aspergillus spp. isolados de grãos de café

    Directory of Open Access Journals (Sweden)

    Marciane Magnani

    2005-01-01

    Full Text Available Some species belonging to the genus Aspergillus are potential producers of ochratoxin A (OA, a mycotoxin with nephrotoxic, immunosuppressive, teratogenic and carcinogenic effects. The aim of the present study was to identify the species of Aspergillus that contaminate the inside of coffee beans collected in the stage of maturation and drying, from 16 producing areas located in the northern region of the State of Paraná, in the South of Brazil. A total of 108 isolates of Aspergillus spp. was identified at the species level, by sequencing the internal transcribed spacer (ITS1-5.8S-ITS2 of ribosomal DNA (rDNA. The results revealed the presence of potentially ochratoxigenic species in 82% of the geographic regions studied, among which Aspergillus niger was the species most frequently detected, followed by A. ochraceus and A. carbonarius. The presence of A. carbonarius in immature coffee fruits harvested from trees is reported for the first time.Algumas espécies pertencentes ao gênero Aspergillus possuem potencial para produção de Ocratoxina A (OA, uma micotoxina de efeitos nefrotóxicos, imunossupressivos, teratogênicos e carcinogênicos. Com o objetivo de identificar as espécies de Aspergillus que contaminam o interior de grãos de café, foram coletadas amostras em diferentes estádios de maturação do produto, em 16 propriedades produtoras do norte do estado do Paraná. Um total de 108 isolados de Aspergillus spp. foram identificados ao nível de espécie, pelo sequenciamento dos espaços internos transcritos (ITS1-5,8S-ITS2 do DNA ribossomal (rDNA. Os resultados revelaram a presença de espécies potencialmente ocratoxigênicas em 82% das regiões analisadas, sendo dentre estas, Aspergillus niger a espécie mais freqüentemente detectada,seguida por A. ochraceus, e A. carbonarius. É relatada pela primeira vez a presença de A. carbonarius em frutos de café coletados na árvore.

  14. Biotransformation of chalcones by the endophytic fungus Aspergillus flavus isolated from Paspalum maritimum trin

    Energy Technology Data Exchange (ETDEWEB)

    Correa, Marivaldo J.C.; Nunes, Fatima M.; Bitencourt, Heriberto R.; Borges, Fabio C.; Guilhon, Giselle M.S.P.; Arruda, Mara S.P.; Marinho, Andrey M. R.; Santos, Alberdan S.; Alves, Claudio N.; Santos, Lourivaldo S., E-mail: lss@ufpa.b [Universidade Federal do Para (IQ/FEQ/UFPA), Belem, PA (Brazil). Inst. de Tecnologia. Faculdade de Engenharia Quimica; Brasil, Davi S.B. [Universidade Federal do Para (PPGQ/IQ/UFPA), Belem, PA (Brazil). Inst. de Quimica. Programa de Pos-Graduacao em Quimica

    2011-07-01

    The fungus Aspergillus flavus isolated as endophytic of the plant Paspalum maritimum Trin. was evaluated for its potential application in biotransformation reactions. The compounds chalcone (1), 3,4,5-trimethoxychalcone (2) and 2,3,4,4'-tetramethoxy chalcone (3) were biotransformed, respectively, in dihydrochalcone (4), 3,4,5-trimethoxydihydrochalcone (5) and 2,3,4,4'-tetramethoxydihydrochalcone (6). The structures were elucidated by spectroscopic methods including 1D and 2D NMR techniques, and MS analysis. The dihydrochalcones 5 and 6 are new compounds. (author)

  15. Novel chromone derivatives from the fungus Aspergillus versicolor isolated from the marine sponge Xestospongia exigua.

    Science.gov (United States)

    Lin, Wenhan; Brauers, Gernot; Ebel, Rainer; Wray, Victor; Berg, Albrecht; Sudarsono; Proksch, Peter

    2003-01-01

    From the marine sponge Xestospongia exigua collected in Indonesia the fungus Aspergillus versicolor was isolated. Following cultivation in a seawater-based medium seven new angular tricyclic chromone derivatives (1-7) were obtained from the mycelia and culture filtrate. Compounds 2-7 contain an additional dihydropyran ring system which is replaced by a pyridine ring in 1. The structures of the new natural products were established on the basis of extensive one- and two-dimensional NMR spectroscopic studies (1H, 13C, COSY, HMQC, HMBC, NOE difference spectra) as well as on mass spectral analysis.

  16. Biotransformation of chalcones by the endophytic fungus Aspergillus flavus isolated from Paspalum maritimum trin

    International Nuclear Information System (INIS)

    Correa, Marivaldo J.C.; Nunes, Fatima M.; Bitencourt, Heriberto R.; Borges, Fabio C.; Guilhon, Giselle M.S.P.; Arruda, Mara S.P.; Marinho, Andrey M. R.; Santos, Alberdan S.; Alves, Claudio N.; Santos, Lourivaldo S.; Brasil, Davi S.B.

    2011-01-01

    The fungus Aspergillus flavus isolated as endophytic of the plant Paspalum maritimum Trin. was evaluated for its potential application in biotransformation reactions. The compounds chalcone (1), 3,4,5-trimethoxychalcone (2) and 2,3,4,4'-tetramethoxy chalcone (3) were biotransformed, respectively, in dihydrochalcone (4), 3,4,5-trimethoxydihydrochalcone (5) and 2,3,4,4'-tetramethoxydihydrochalcone (6). The structures were elucidated by spectroscopic methods including 1D and 2D NMR techniques, and MS analysis. The dihydrochalcones 5 and 6 are new compounds. (author)

  17. Isolation of azole-resistant Aspergillus fumigatus from the environment in the south-eastern USA.

    Science.gov (United States)

    Hurst, Steven F; Berkow, Elizabeth L; Stevenson, Katherine L; Litvintseva, Anastasia P; Lockhart, Shawn R

    2017-09-01

    Azole resistance in isolates of the fungus Aspergillus fumigatus has been associated with agricultural use of azole fungicides. Environmental isolation of resistant isolates has been reported in Asia, Africa, Europe and South America. To determine whether A. fumigatus isolates containing TR34/L98H or TR46/Y121F/T289A can be found in fields in the USA treated with agricultural azoles. Crop debris was collected and screened for A. fumigatus. All A. fumigatus isolates were screened for azole resistance. The CYP51A gene of azole-resistant isolates was sequenced. The population structure of a subset of isolates was determined using microsatellite typing. This article identifies azole-resistant A. fumigatus isolates containing the TR34/L98H mutation in an experimental peanut field that had been treated with azole fungicides. These findings suggest the development of resistance to azole antifungals in A. fumigatus may be present where agricultural azoles are used in the USA. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy 2017. This work is written by US Government employees and is in the public domain in the US.

  18. Pharmacodynamics of Voriconazole against Wild-Type and Azole-Resistant Aspergillus flavus Isolates in a Nonneutropenic Murine Model of Disseminated Aspergillosis.

    Science.gov (United States)

    Rudramurthy, Shivaprakash M; Seyedmousavi, Seyedmojtaba; Dhaliwal, Manpreet; Chakrabarti, Arunaloke; Meis, Jacques F; Mouton, Johan W

    2017-01-01

    Invasive aspergillosis (IA) due to Aspergillus flavus is associated with high mortality. Although voriconazole (VRC) is widely recommended as the first-line treatment for IA, emergence of azole resistance in Aspergillus spp. is translating to treatment failure. We evaluated the efficacy of voriconazole in a nonneutropenic murine model of disseminated A. flavus infection using two voriconazole-resistant isolates (one harboring the Y319H substitution in the cyp51C gene) and two wild-type isolates without mutations. All isolates exhibited a dose-response relationship, and voriconazole treatment improved mouse survival in a dose-dependent manner. At 40 mg/kg of body weight, 100% efficacy was observed for 1 susceptible isolate and 1 resistant isolate (with mutation), whereas for another susceptible isolate and resistant isolate (without mutation), survival rates were 81% and 72%, respectively. The Hill equation with a variable slope fitted the relationship between the area under the concentration-time curve (AUC)/MIC ratio and 14-day survival well for each strain. An F test showed the 50% effective doses to be significantly different from each other (P = 0.0023). However, contrary to expectation, there was a significant difference in exposure-response relationships between strains, and it appeared that the susceptible strains required a relatively higher exposure than the resistant ones to result in the same treatment effect, the 50% effective pharmacokinetic/pharmacodynamic (PK/PD) index (EI 50 ) required being negatively and log-linearly related to the MIC (P = 0.04). We conclude that the efficacy of voriconazole depended on drug exposure and the voriconazole MIC of the isolates, but lower exposures are required for strains with higher MICs. These findings may have profound significance in clinical practice with respect to dosing and drug choice. Copyright © 2016 American Society for Microbiology.

  19. Potent synergistic in vitro interaction between nonantimicrobial membrane-active compounds and itraconazole against clinical isolates of Aspergillus fumigatus resistant to itraconazole.

    NARCIS (Netherlands)

    Afeltra, J.; Vitale, R.G.; Mouton, J.W.; Verweij, P.E.

    2004-01-01

    To develop new approaches for the treatment of invasive infections caused by Aspergillus fumigatus, the in vitro interactions between itraconazole (ITZ) and seven different nonantimicrobial membrane-active compounds--amiodarone (AMD), amiloride, lidocaine, lansoprazole (LAN), nifedipine (NIF),

  20. Steroids and an oxylipin from an algicolous isolate of Aspergillus flavus.

    Science.gov (United States)

    Qiao, Ming-Feng; Ji, Nai-Yun; Miao, Feng-Ping; Yin, Xiu-Li

    2011-06-01

    A new oxylipin, (8E,12Z)-10,11-dihydroxyoctadeca-8,12-dienoic acid (1), a new steroid, 3β,4α-dihydroxy-26-methoxyergosta-7,24(28)-dien-6-one (2), and four known steroids, episterol (3), (22E,24R)-ergosta-7,22-dien-3β,5α,6α-triol (4), (22E,24R)-ergosta-5,22-dien-3β-ol (5), and (22E,24R)-ergosta-4,6,8(14),22-tetraen-3-one (6), were isolated from the cultures of Aspergillus flavus, an endophytic fungus isolated from the marine red alga Corallina officinalis. Their structures and relative stereochemistry were elucidated by 1D, 2D NMR and mass spectroscopic techniques. 1 and 2 exhibited low activity to inhibit acetylcholinesterase and no activity against plant pathogenic fungi Colletotrichum lagenarium and Fusarium oxysporum. Copyright © 2011 John Wiley & Sons, Ltd.

  1. Optimal pcr primers for rapid and accurate detection of Aspergillus flavus isolates.

    Science.gov (United States)

    Al-Shuhaib, Mohammed Baqur S; Albakri, Ali H; Alwan, Sabah H; Almandil, Noor B; AbdulAzeez, Sayed; Borgio, J Francis

    2018-03-01

    Aspergillus flavus is among the most devastating opportunistic pathogens of several food crops including rice, due to its high production of carcinogenic aflatoxins. The presence of these organisms in economically important rice strip farming is a serious food safety concern. Several polymerase chain reaction (PCR) primers have been designed to detect this species; however, a comparative assessment of their accuracy has not been conducted. This study aims to identify the optimal diagnostic PCR primers for the identification of A. flavus, among widely available primers. We isolated 122 A. flavus native isolates from randomly collected rice strips (N = 300). We identified 109 isolates to the genus level using universal fungal PCR primer pairs. Nine pairs of primers were examined for their PCR diagnostic specificity on the 109 isolates. FLA PCR was found to be the optimal PCR primer pair for specific identification of the native isolates, over aflP(1), aflM, aflA, aflD, aflP(3), aflP(2), and aflR. The PEP primer pair was found to be the most unsuitable for A. flavus identification. In conclusion, the present study indicates the powerful specificity of the FLA PCR primer over other commonly available diagnostic primers for accurate, rapid, and large-scale identification of A. flavus native isolates. This study provides the first simple, practical comparative guide to PCR-based screening of A. flavus infection in rice strips. Copyright © 2018 Elsevier Ltd. All rights reserved.

  2. Genetic variability of Aspergillus flavus isolates from a Mississippi corn field.

    Science.gov (United States)

    Solorzano, Cesar D; Abbas, Hamed K; Zablotowicz, Robert M; Chang, Perng-Kuang; Jones, Walker A

    2014-01-01

    A nontoxigenic Aspergillus flavus strain, K49, is currently being tested as a biological control agent in corn fields in the Mississippi Delta. However, little is known about the overall genetic diversity of A. flavus from year to year in corn fields and specifically in Mississippi. Our objective was to assess the genetic variability of A. flavus isolates from different seasons, inoculum sources, and years, from a no-till corn field. Of the 175 A. flavus isolates examined, 74 and 97 had the typical norB-cypA type I (1.5 kb) and type II (1.0 kb) deletion patterns, respectively. Variability in the sequence of the omtA gene of the majority of the field isolates (n = 118) was compared to strain K49. High levels of haplotypic diversity (24 omtA haplotypes; Hd = 0.61 ± 0.04) were found. Among the 24 haplotypes, two were predominant, H1 (n = 71), which consists of mostly toxigenic isolates, and H49 (n = 18), which consists of mostly atoxigenic isolates including K49. Toxigenic isolates were prevalent (60%) in this natural population. Nonetheless, about 15% of the population likely shared the same ancestral origin with K49. This study provides valuable information on the diversity of A. flavus. This knowledge can be further used to develop additional biological control strains.

  3. Genetic Variability of Aspergillus flavus Isolates from a Mississippi Corn Field

    Directory of Open Access Journals (Sweden)

    Cesar D. Solorzano

    2014-01-01

    Full Text Available A nontoxigenic Aspergillus flavus strain, K49, is currently being tested as a biological control agent in corn fields in the Mississippi Delta. However, little is known about the overall genetic diversity of A. flavus from year to year in corn fields and specifically in Mississippi. Our objective was to assess the genetic variability of A. flavus isolates from different seasons, inoculum sources, and years, from a no-till corn field. Of the 175 A. flavus isolates examined, 74 and 97 had the typical norB-cypA type I (1.5 kb and type II (1.0 kb deletion patterns, respectively. Variability in the sequence of the omtA gene of the majority of the field isolates (n=118 was compared to strain K49. High levels of haplotypic diversity (24 omtA haplotypes; Hd = 0.61 ± 0.04 were found. Among the 24 haplotypes, two were predominant, H1 (n=71, which consists of mostly toxigenic isolates, and H49 (n=18, which consists of mostly atoxigenic isolates including K49. Toxigenic isolates were prevalent (60% in this natural population. Nonetheless, about 15% of the population likely shared the same ancestral origin with K49. This study provides valuable information on the diversity of A. flavus. This knowledge can be further used to develop additional biological control strains.

  4. Evaluation of the in vitro activity of isavuconazole and comparator voriconazole against 2635 contemporary clinical Candida and Aspergillus isolates

    DEFF Research Database (Denmark)

    Astvad, K. M.T.; Hare, R. K.; Arendrup, M. C.

    2017-01-01

    Objective The in vitro activity of isavuconazole was determined for 1677 Candida and 958 Aspergillus isolates from 2012 to 2014 with voriconazole as comparator. Methods Aspergillus isolates were screened for resistance using azole-agar. Aspergillus isolates that screened positive and all Candida...

  5. Voriconazole Exposure and Risk of Cutaneous Squamous Cell Carcinoma, Aspergillus Colonization, Invasive Aspergillosis and Death in Lung Transplant Recipients.

    Science.gov (United States)

    Mansh, M; Binstock, M; Williams, K; Hafeez, F; Kim, J; Glidden, D; Boettger, R; Hays, S; Kukreja, J; Golden, J; Asgari, M M; Chin-Hong, P; Singer, J P; Arron, S T

    2016-01-01

    Voriconazole is a triazole antifungal used to prevent and treat invasive fungal infections after lung transplantation, but it has been associated with an increased risk of developing cutaneous squamous cell carcinoma (SCC). Despite widespread use, there are no clear guidelines for optimal prophylactic regimens that balance the competing risks and benefits. We conducted a retrospective cohort study of all lung transplant recipients at the University of California, San Francisco, who were transplanted between October 1991 and December 2012 (n = 455) to investigate whether voriconazole exposure affected development of SCC, Aspergillus colonization, invasive aspergillosis and all-cause mortality. Voriconazole exposure was associated with a 73% increased risk of developing SCC (hazard ratio [HR] 1.73; 95% confidence interval [CI]: 1.04-2.88; p = 0.03), with each additional 30-day exposure at the standard dose increasing the risk by 3.0% (HR 1.03; 95% CI: 1.02-1.04; p Voriconazole exposure reduced risk of Aspergillus colonization by 50% (HR 0.50; 95% CI: 0.34-0.72; p Voriconazole exposure significantly reduced all-cause mortality among subjects who developed Aspergillus colonization (HR 0.34; 95% CI: 0.13-0.91; p = 0.03) but had no significant impact on those without colonization. Physicians should consider patient-specific factors that modify the potential risks and benefits of voriconazole for the care of lung transplant recipients. © Copyright 2015 The American Society of Transplantation and the American Society of Transplant Surgeons.

  6. Plastic degrading fungi Trichoderma viride and Aspergillus nomius isolated from local landfill soil in Medan

    Science.gov (United States)

    Munir, E.; Harefa, R. S. M.; Priyani, N.; Suryanto, D.

    2018-03-01

    Plastic is a naturally recalcitrant polymer, once it enters the environment, it will remain there for many years. Accumulation of plastic as wastes in the environment poses a serious problem and causes an ecological threat. Alternative strategies to reduce accumulation of plastic wastes have been initiated and implemented from a different aspect including from microbiological view point. The study to obtain potential fungi in degrading plastic molecule has been initiated in our laboratory. Low density polyethylene (LDPE) plastic was used as a tested material. Candidate fungi were isolated from local landfill soil. The fungi were cultured in mineral salt medium broth containing LDPE powder. Two of nine isolates showed best growth response in broth media containing LDPE. These isolates (RH03 and RH06) were used in degradation test. Results showed that isolate RH03 and RH06 reduced the weight of LDPE film by 5.13% and 6.63%, respectively after 45 days of cultivation. The tensile strength of treated film even reduced significantly by 58% and 40% of each isolate. Analyses of electron micrograph exhibited grove ands rough were formed on the surface of LDPE film. These were not found in the untreated film. Furthermore, molecular analysis through polymerase chain reaction and DNA sequencing indicated that RH03 is Trichoderma viride and RH06 is Aspergillus nomius with 97% and 96% similarities, respectively.

  7. Radiosensitivity of toxigenic Aspergillus isolated from spices and destruction of aflatoxins by gamma-irradiation

    Science.gov (United States)

    Kume, Tamikazu; Ito, Hitoshi; Soedarman, Harsono; Ishigaki, Isao

    Radiosensitivities of Aspergillus flavus var columnaris isolated from spices were investigated. The D10 values and induction doses were 267-293 Gy and 75-165 Gy in wet conditions, respectively. In dry conditions, the survival curves were exponential and D10 values were 538-600 Gy. The survival curves of standard strain of A. parasiticus IFO 30179 were similar both in wet and dry conditions. The necessary dose of 8 kGy for the destruction of these toxigenic Aspergillus was calculated from these values. Two of 11 strains of A. flavus var columnaris produced aflatoxins and the content of B 1 was especially high. In the study of irradiation effect on aflatoxins produced on polished rice, aflatoxins G 1 and B 1 were more radiosensitive than G 2 and B 2. However, these aflatoxins were very stable to radiation and the dose required for destruction was found to be more than 500 kGy. It is therfore concluded that the decontamination of molds by irradiation is necessary prior to their production of aflatoxins.

  8. Case report of a new pathogenic variant of Aspergillus fumigates isolated from Hipposideros cervinus (Chiroptera: Hipposideridae in Sarawak, Malaysia

    Directory of Open Access Journals (Sweden)

    S.S.J. Seelan

    2009-03-01

    Full Text Available First record of new Aspergillus fumigatus variant (UNIMAS F009 was reported from the ears of bats at Kubah National Park, Borneo, Malaysia. Morphological characterization of this isolate showed some differences in terms of their growth rate, colony color, size of conidia and pigmentation on different media.

  9. Effects of Hydrogen Peroxide on Different Toxigenic and Atoxigenic Isolates of Aspergillus flavus

    Directory of Open Access Journals (Sweden)

    Jake C. Fountain

    2015-08-01

    Full Text Available Drought stress in the field has been shown to exacerbate aflatoxin contamination of maize and peanut. Drought and heat stress also produce reactive oxygen species (ROS in plant tissues. Given the potential correlation between ROS and exacerbated aflatoxin production under drought and heat stress, the objectives of this study were to examine the effects of hydrogen peroxide (H2O2-induced oxidative stress on the growth of different toxigenic (+ and atoxigenic (− isolates of Aspergillus flavus and to test whether aflatoxin production affects the H2O2 concentrations that the isolates could survive. Ten isolates were tested: NRRL3357 (+, A9 (+, AF13 (+, Tox4 (+, A1 (−, K49 (−, K54A (−, AF36 (−, and Aflaguard (−; and one A. parasiticus isolate, NRRL2999 (+. These isolates were cultured under a H2O2 gradient ranging from 0 to 50 mM in two different media, aflatoxin-conducive yeast extract-sucrose (YES and non-conducive yeast extract-peptone (YEP. Fungal growth was inhibited at a high H2O2 concentration, but specific isolates grew well at different H2O2 concentrations. Generally the toxigenic isolates tolerated higher concentrations than did atoxigenic isolates. Increasing H2O2 concentrations in the media resulted in elevated aflatoxin production in toxigenic isolates. In YEP media, the higher concentration of peptone (15% partially inactivated the H2O2 in the media. In the 1% peptone media, YEP did not affect the H2O2 concentrations that the isolates could survive in comparison with YES media, without aflatoxin production. It is interesting to note that the commercial biocontrol isolates, AF36 (−, and Aflaguard (−, survived at higher levels of stress than other atoxigenic isolates, suggesting that this testing method could potentially be of use in the selection of biocontrol isolates. Further studies will be needed to investigate the mechanisms behind the variability among isolates with regard to their degree of oxidative stress

  10. Geographically predominant genotypes of Aspergillus terreus species complex in Austria: s microsatellite typing study

    NARCIS (Netherlands)

    Lackner, M.; Coassin, S.; Haun, M.; Binder, U.; Kronenberg, F.; Haas, H. de; Jank, M.; Maurer, E.; Meis, J.F.; Hagen, F.; Lass-Florl, C.

    2016-01-01

    Aspergillus terreus species complex is recognized as a frequent agent of invasive aspergillosis in Tyrol. The reason for this specific epidemiological situation is unclear. Aspergillus terreus strains isolated from environmental and clinical sources were genotyped using a novel panel of short tandem

  11. Anti - Aspergillus flavus Activity of Lactobacillus plantarum and Pediococcus acidilactici Isolated from Breast Milk

    Directory of Open Access Journals (Sweden)

    Sedighi-Khavidak Samaneh

    2016-12-01

    Full Text Available The breast milk is not only the best and optimal natural source of essential nutrients for babies, but also provides a broad range of bioactive compounds, as well as its own microbiota, therefore helps to protect body against many diseases too. Lactic acid bacteria (LAB are the major group of the breast milk microbiota and acts as biocontrol agent in milk. In this study LAB were isolated from the breast milk. Their antifungal activity against aflatoxin producing Aspergillus flavus was investigated with four methods: Study of the LAB isolates influence on biomass production of A. flavus in MRS medium, determination of their inhibitory activity against A. flavus on PDA medium, dual culture overlay assay and finally, effect of LAB cell-free supernatants (CFS against mycelia growth. The CFS also was analyzed by HPLC and titration experiments. The results indicated that seven isolated LAB had good inhibitory activity against A. flavus growth. Four isolates of LAB were identified as Pediococcus acidilactici and three of them were identified as Lactobacillus plantarum by 16s rDNA sequencing assay. The cell-free supernatants of L. plantarum RS2 expressed strong anti A. flavus activity. Analyzing of their CFS showed that, this strain had significant amounts of propionic acid and hydrogen peroxide in the CFS that might explain the high antifungal activity.

  12. Effect of Plant Essential Oils and Gamma Irradiation on Growth and Aflatoxin Production by Aspergillus Flavus Isolated from Wheat Grains

    International Nuclear Information System (INIS)

    Salem, E.A.; Shalaby, Kh.

    2016-01-01

    The antifungal potential of essential oils of Thyme (Thymus vulgaris L.) and camphor ( Eucalyptus rostrata L.) was determined on Aspergillus flavus link isolated from wheat grains on Potato dextrose agar (PDA). They inhibited completely mycelia growth of the fungus at 1000 and 2000 ppm, and prevented aflatoxin production at sub lethal dose 500 and 1000 ppm respectively. Gamma radiation was used to control mycelia growth of Aspergillus flavus Link and inhibiting aflatoxin production. A dose level of 3.5 KGy gamma radiation prevented the fungal growth and aflatoxin production by A. flavus link, where a dose of 2.5 K Gy ( the sub lethal dose) prevented about 85% of aflatoxin production

  13. Cyclo-(trp-phe diketopiperazines from the endophytic fungus Aspergillus versicolor isolated from Piper aduncum

    Directory of Open Access Journals (Sweden)

    Juliana R. Gubiani

    Full Text Available Six known compounds, three peptide derivatives: cyclo-(tryptophyl-phenylalanyl (2, diketopiperazine dimer WIN 64821 (3 and 3-hydroxy-15H-tryptophenaline (4, one adenine derivative: 2-hydroxy-6-N-isopentenyl-adenine (5, one phtalide derivative: 4-methoxyphtalide (1 and one benzoic acid derivative: 3-hydroxy-4-(1-hydroxy-1,5-dimethyl-hexyl benzoic acid (6, were isolated from the ethyl acetate extract of the endophytic fungus Aspergillus versicolor associated with the Piper aduncum plant. Their structures were determined on the basis of detailed interpretation of 1 D and 2D NMR spectra and in comparison with works reported in the literature. This paper, in effect, deals with the first report of these compounds in A. versicolor.

  14. Isolation and genetic analysis of Aspergillus niger mutants with reduced extracellular glucoamylase

    International Nuclear Information System (INIS)

    Valent, G.U.; Calil, M.R.; Bonatelli Junior, R.

    1992-01-01

    Mutants with impaired production of extracellular glucoamylase were isolated at a high frequency (2% of survivors) from an Aspergillus niger strain treated with UV light. These were designated as low glucoamylase producers (lgp, up to 30% of the parental yield) and medium producers (mgp, a 35 to 50% decrease in enzyme level). All the mutants were shown to be recessive; one strain segregated two unlinked genes. Complementation tests, and segregation from heterozygous diploid, suggested at least three to four unlinked genes, each able to impair glucoamylase production. There is evidence of a single structural gene for glucoamylase in A. niger. Therefore, as production of extracellular enzymes is normally the final result of several steps at intracellular and membrane levels, including regulation of enzyme synthesis, we suggest intergenic interaction that controls extracellular enzyme accumulation and that mutation in any of these genes would result in impaired production. (author)

  15. Isolation of marine fungi Aspergillus sp. and its in vitro antifouling activity against marine bacteria.

    Science.gov (United States)

    Thiyagarajan, Santhananmari; Bavya, Manoharan; Jamal, Alruwaili

    2016-09-01

    Biofouling is considered as a main issue of concern in aquatic environment causing severe economic loss and pollution. The aim of the present study was to isolate marine fungus antagonistic to biofouling bacteria and to define antifouling compounds present in it. Using standard plate method five predominant biofouling bacteria viz., Methylococcus sp., Flavobacterium sp., Marinococcus sp., Serratia sp. and Pseudomonas sp. were isolated from marine solid substances on Zobell's agar. Tolerance range of these bacteria to NaCl was 2-10%. Isolation of fungi from mangrove and estuarine sediments and their screening identified Aspergillus sp. EF4 as a potential isolate. This isolate caused inhibition of all the five test bacterial cultures measuring zone diameters respectively of 11, 16, 12, 13 and 11mm.? Subsequent to submerged fermentation using shaking flask method this fungus produced bioactive compounds within 5 days. The culture parameters optimized were raffinose as carbon source, yeast extract as lone nitrogen source, pH up to 9.0 and temperature up to 40?C. Antifouling compounds of culture filtrate were separated and detected by a three-step procedure involving thin layer chromatography, bioautography and preparative TLC. The in vitro assay involving glass slide-wooden stick-biofilm method revealed that these compounds could cause inhibition and destruction of bacteria to an extent of 2.16 x 104 CFU ml-1 and 2.46 x 104 CFU ml-1 respectively while growth of bacteria in control beaker was enumerated to be 4.41 x 104 CFU ml-1. High performance liquid chromatography of culture filtrate indicated probable principal antifouling compound as Fumonisin B2. Isolation of antagonistic marine fungus from Indian coast and detection of its antifouling compound would help in planning effective strategies for controlling biofouling in marine environment.

  16. Production of catalases by Aspergillus niger isolates as a response to pollutant stress by heavy metals

    Energy Technology Data Exchange (ETDEWEB)

    Buckova, M.; Godocikova, J.; Simonovicova, A.; Polek, B. [Slovakian Academy of Science, Bratislava (Slovakia)

    2005-04-15

    Isolates of Aspergillus niger, selected from the coal dust of a mine containing arsenic (As; 400 mg/kg) and from the river sediment of mine surroundings (As, 1651 mg/kg, Sb, 362 mg/kg), growing in minimal nitrate medium in the phase of hyphal development and spore formation, exhibited much higher levels of total catalase activity than the same species from the culture collection or a culture adapted to soil contaminated with As (5 mg/L). Electrophoretic resolution of catalases in cell-free extracts revealed three isozymes of catalases and production of individual isozymes was not significantly affected by stress environments. Exogenously added stressors (As{sup 5+}, Cd{sup 2+}, Cu{sup 2+}) at final concentrations of 25 and 50 mg/L and H{sub 2}O{sub 2} (20 or 40 m(M)) mostly stimulated production of catalases only in isolates from mines surroundings, and H{sub 2}O{sub 2} and Hg{sup 2+} caused the disappearance of the smallest catalase I. Isolates exhibited a higher tolerance of the toxic effects of heavy metals and H{sub 2}O{sub 2}, as monitored by growth, than did the strain from the culture collection.

  17. Optimization of pectinase enzyme production in Aspergillus fumigatus isolated from rotten fruits

    Directory of Open Access Journals (Sweden)

    2015-12-01

    Full Text Available Introduction: Pectinase is one of the most important industrial enzymes which was isolated from a wide variety of microorganisms such as bacteria and filamentous fungi. This enzyme has been usually used in the juice and textile industry. In this study, the isolation and optimization of pectinase-producing fungi on decaying rotten fruits were studied. Materials and methods: Isolation and screening of pectinase producing fungi have been done by plate culture on pectin medium and staining with Lugol's iodine solution. The best strain was identified by method of Pitt and Hocking as Aspergillus fumigates. The enzyme production was optimized by application of the factorial design which involves five factors, each at three levels. Five factors were carbon sources (whey, sugar, stevia and ammonium sulfate, manganese sulfate, temperature, and pH. Pectinase concentration was measured by the Miller method. Results: The results showed that the optimum condition for enzyme production was at 32 °C, PH = 6 , 3g / L manganese sulfate, 2.75g / L of ammonium sulfate, 10g / L of each carbon source (whey, stevia, and glucose. Optimum of enzyme production was observed in the presence of 1.328 mg / ml of glucose. Molecular weight of enzyme was obtained about 40 kDa by SDS-PAGE. Discussion and conclusion: The results demonstrated that this strain could grow in a wide range of carbon sources, PH and temperature. This study indicates that this strain is a good candidate for use in industrial application.

  18. Aspergillus niger causing tracheobronchitis and invasive pulmonary aspergillosis in a lung transplant recipient: case report Aspergillus niger causando traqueobronquite e aspergilose pulmonar invasiva em transplantado de pulmão: relato de caso

    Directory of Open Access Journals (Sweden)

    Melissa Orzechowski Xavier

    2008-04-01

    Full Text Available A case of invasive aspergillosis caused by Aspergillus niger in a lung transplant recipient is described. The patient presented hyperglycemia starting postoperatively, with other complications such as cytomegalovirus infection. The associated predisposing factors and other implications are discussed. Aspergillus niger seems to be a fungal species of low virulence that requires the presence of a severely immunosuppressed host to cause invasive disease.Descreve-se um caso de aspergilose invasiva causada por Aspergillus niger em um paciente transplantado de pulmão com quadros hiperglicêmicos desde o pós-operatório e outras complicações como infecção por citomegalovírus. Os fatores predisponentes associados e outras implicações são discutidos. Aspergillus niger parece ser uma espécie fúngica de baixa virulência, necessitando a presença de um hospedeiro gravemente imunodeprimido para causar doença invasiva.

  19. Assay of antioxidant potential of two Aspergillus isolates by different methods under various physio-chemical conditions.

    Science.gov (United States)

    Arora, Daljit Singh; Chandra, Priyanka

    2010-07-01

    The objective of this work was to screen fungi isolated from soil of different areas of Punjab, India for antioxidant activity by dot blot assay and around 45% of fungal isolates demonstrated antioxidant potential. Two selected strains of Aspergillus spp (Aspergillus PR78 and Aspergillus PR66) showing quantitatively best antioxidant activity by DPPH assay were further tested for their reducing power, ferrous ion and nitric oxide ion scavenging activity, FRAP assay and total phenolic content. Different physio-chemical parameters were optimized for enhancement of the activity. This revealed stationary culture grown for 10 days at 25 (o)C at pH 7 to be the best for antioxidant activity. Sucrose in the medium as carbon source resulted in highest antioxidant activity. Sodium nitrate, yeast extract, and peptone were good sources of nitrogen but sodium nitrate was the best among these. The extraction of the broth culture filtrates with different solvents revealed ethyl acetate extract to possess the best antioxidant activity. The activity as expressed by ethyl acetate extract of Aspergillus PR78 was equally effective as that of commonly used antioxidant standard, ascorbic acid.

  20. Assay of antioxidant potential of two Aspergillus isolates by different methods under various physio-chemical conditions

    Directory of Open Access Journals (Sweden)

    Daljit Singh Arora

    2010-10-01

    Full Text Available The objective of this work was to screen fungi isolated from soil of different areas of Punjab, India for antioxidant activity by dot blot assay and around 45% of fungal isolates demonstrated antioxidant potential. Two selected strains of Aspergillus spp (Aspergillus PR78 and Aspergillus PR66 showing quantitatively best antioxidant activity by DPPH assay were further tested for their reducing power, ferrous ion and nitric oxide ion scavenging activity, FRAP assay and total phenolic content. Different physio-chemical parameters were optimized for enhancement of the activity. This revealed stationary culture grown for 10 days at 25ºC at pH 7 to be the best for antioxidant activity. Sucrose in the medium as carbon source resulted in highest antioxidant activity. Sodium nitrate, yeast extract, and peptone were good sources of nitrogen but sodium nitrate was the best among these. The extraction of the broth culture filtrates with different solvents revealed ethyl acetate extract to possess the best antioxidant activity. The activity as expressed by ethyl acetate extract of Aspergillus PR78 was equally effective as that of commonly used antioxidant standard, ascorbic acid.

  1. Isolation of Aspergillus species from Nasal Cavity and Bedroom of Healthy Volunteers and Patients with Allergic Rhinitis in Mashhad, Iran

    Directory of Open Access Journals (Sweden)

    Samaneh Eidi

    2014-12-01

    Full Text Available Background: The purpose of this study was to investigate the presence, frequency and comparison of Aspergillus spp. in nasal cavity and bedroom of healthy volunteers and patients with allergic rhinitis. Materials and Methods: In this cross-sectional study, a group of patients with allergic rhinitis (N=50 were selected based on positive skin prick test. Healthy volunteers were chosen to be in the comparison group by matching in age, gender, and no history of respiratory system disease. Samples from nasal cavity and different parts of bedroom were collected and cultured. Cultured Aspergillus spp. was identified by standard mycological techniques. Results: The most common species isolated from all samples of healthy volunteers was A. flavus (88%, followed by A. niger (76% and A. fumigatus (74%. A. flavus (56% was the predominant species isolated from all samples of patients, followed by A. niger (34% and A. fumigatus (6%. Conclusion: A. flavus was the most prevalent species of Aspergillus both healthy volunteers and patients. The presence of Aspergillus in homes does not necessarily imply a cause and effect relationship with illness, but we speculate that A. flavus may be a major source of aeroallergens along with A. niger and A. fumigatus; and should alert physicians and healthcare professionals to do more vigorous environmental testing.

  2. Induction of Pectinase Hyper Production by Multistep Mutagenesis Using a Fungal Isolate--Aspergillus flavipes.

    Science.gov (United States)

    Akbar, Sabika; Prasuna, R Gyana; Khanam, Rasheeda

    2014-04-01

    Aspergillus flavipes, a slow growing pectinase producing ascomycete, was isolated from soil identified and characterised in the previously done preliminary studies. Optimisation studies revealed that Citrus peel--groundnut oil cake [CG] production media is the best media for production of high levels of pectinase up to 39 U/ml using wild strain of A. flavipes. Strain improvement of this isolated strain for enhancement of pectinase production using multistep mutagenesis procedure is the endeavour of this project. For this, the wild strain of A. flavipes was treated with both physical (UV irradiation) and chemical [Colchicine, Ethidium bromide, H2O2] mutagens to obtain Ist generation mutants. The obtained mutants were assayed and differentiated basing on pectinase productivity. The better pectinase producing strains were further subjected to multistep mutagenesis to attain stability in mutants. The goal of this project was achieved by obtaining the best pectinase secreting mutant, UV80 of 45 U/ml compared to wild strain and sister mutants. This fact was confirmed by quantitatively analysing 3rd generation mutants obtained after multistep mutagenesis.

  3. Antifungal activity of lemon, eucalyptus, thyme, oregano, sage and lavender essential oils against Aspergillus niger and Aspergillus tubingensis isolated from grapes

    Directory of Open Access Journals (Sweden)

    Miroslava Císarová

    2016-01-01

    Full Text Available Today, it is very important to find out the protection of products of natural origin as an alternative to synthetic fungicides. The promising alternative is the use of the essential oils (EOs. Essential oils from plants have great potential as a new source of fungicide to control the pathogenic fungi.The main objective of this study was evaluation of the antifungal activity of lemon (Citrus lemon L., eucalyptus (Eucalyptus globulus LABILL., thyme (Thymus vulgaris L., oregano (Origanum vulgare L. sage (Salvia officinalis L. and lavender (Lavandula angustifolia MILLER. EOs against Aspergillus niger and Aspergillus tubingensis isolated from grapes and their ability to affect the growth. It was tested by using the vapor contact with them. At first both tested isolates were identified by using PCR method. Sequence data of 18S rRNA supported the assignment of these isolates to the genus Aspergillus and species A. niger (ITS region: KT824061; RPB2: KT824060 and A. tubingensis (ITS region: KT824062; RPB2: KT824059. Second, EO antifungal activity was evaluated. The effect of the EO volatile phase was confirmed to inhibit growth of A. niger and A tubingensis. EOs were diluted in DMSO (dimethyl sulfoxide final volume of 100 μL. Only 50 μL this solution was distributed on a round sterile filter paper (1 x 1 cm by micropipette, and the paper was placed in the center of the lid of Petri dishes. Dishes were kept in an inverted position. The essential oils with the most significant activity were determined by method of graded concentration of oils - minimum inhibitory doses (MIDs. The most effective tested EOs were oregano and thyme oils, which totally inhibited growth of tested isolates for all days of incubation at 0.625 μL.cm-3 (in air with MFDs 0.125 μL.cm-3 (in air. Lavender EO was less active aginst tested strains (MIDs 0.313 μL.cm-3. The results showed that the tested EOs had antifungal activity, except lemon and eucalyptus. Sage EO was the only

  4. Expression of Genes by Aflatoxigenic and Nonaflatoxigenic Strains of Aspergillus flavus Isolated from Brazil Nuts.

    Science.gov (United States)

    Baquião, Arianne Costa; Rodriges, Aline Guedes; Lopes, Evandro Luiz; Tralamazza, Sabina Moser; Zorzete, Patricia; Correa, Benedito

    2016-08-01

    The aims of the present study were to monitor the production of aflatoxin B1 (AFB1) and mycelial growth, and to evaluate the expression of genes directly and indirectly involved in the biosynthesis of aflatoxins by Aspergillus flavus isolated from Brazil nuts. Six previously identified A. flavus strains were grown on coconut agar at 25°C for up to 10 days. Mycotoxins were separated by high-performance liquid chromatography and fungal growth was measured daily using the diametric mycelial growth rate. Transcriptional analysis was performed by real-time polymerase chain reaction (PCR) after 2 and 7 d of incubation using specific primers (aflR, aflD, aflP, lipase, metalloprotease, and LaeA). Three (50%) of the six A. flavus isolates produced AFB1 (ICB-1, ICB-12, and ICB-54) and three (50%) were not aflatoxigenic (ICB-141, ICB-161, and ICB-198). Aflatoxin production was observed from d 2 of incubation (1.5 ng/g for ICB-54) and increased gradually with time of incubation until d 10 (15,803.6 ng/g for ICB-54). Almost all A. flavus isolates exhibited a similar gene expression pattern after 2 d of incubation (p > 0.10). After 7 d of incubation, the LaeA (p aflatoxin production in A. flavus and that overexpression of aflR could affect the transcriptional and aflatoxigenic pattern (ICB-54). Elucidation of the molecular mechanisms that regulate the secondary metabolism of toxigenic fungi may permit the rational silencing of the genes involved and consequently the programmed inhibition of aflatoxin production. Knowledge of the conditions, under which aflatoxin genes are expressed, should contribute to the development of innovative and more cost-effective strategies to reduce and prevent aflatoxin contamination in Brazil nuts.

  5. Molecular detection and species-specific identification of medically important Aspergillus species by real-time PCR in experimental invasive pulmonary aspergillosis.

    Science.gov (United States)

    Walsh, Thomas J; Wissel, Mark C; Grantham, Kevin J; Petraitiene, Ruta; Petraitis, Vidmantas; Kasai, Miki; Francesconi, Andrea; Cotton, Margaret P; Hughes, Johanna E; Greene, Lora; Bacher, John D; Manna, Pradip; Salomoni, Martin; Kleiboeker, Steven B; Reddy, Sushruth K

    2011-12-01

    Diagnosis of invasive pulmonary aspergillosis (IPA) remains a major challenge to clinical microbiology laboratories. We developed rapid and sensitive quantitative PCR (qPCR) assays for genus- and species-specific identification of Aspergillus infections by use of TaqMan technology. In order to validate these assays and understand their potential diagnostic utility, we then performed a blinded study of bronchoalveolar lavage (BAL) fluid specimens from well-characterized models of IPA with the four medically important species. A set of real-time qPCR primers and probes was developed by utilizing unique ITS1 regions for genus- and species-specific detection of the four most common medically important Aspergillus species (Aspergillus fumigatus, A. flavus, A. niger, and A. terreus). Pan-Aspergillus and species-specific qPCRs with BAL fluid were more sensitive than culture for detection of IPA caused by A. fumigatus in untreated (P < 0.0007) and treated (P ≤ 0.008) animals, respectively. For infections caused by A. terreus and A. niger, culture and PCR amplification from BAL fluid yielded similar sensitivities for untreated and treated animals. Pan-Aspergillus PCR was more sensitive than culture for detection of A. flavus in treated animals (P = 0.002). BAL fluid pan-Aspergillus and species-specific PCRs were comparable in sensitivity to BAL fluid galactomannan (GM) assay. The copy numbers from the qPCR assays correlated with quantitative cultures to determine the pulmonary residual fungal burdens in lung tissue. Pan-Aspergillus and species-specific qPCR assays may improve the rapid and accurate identification of IPA in immunocompromised patients.

  6. Involvement of the opportunistic pathogen Aspergillus tubingensis in osteomyelitis of the maxillary bone: A case report

    NARCIS (Netherlands)

    Bathoorn, E.; Escobar Salazar, N.; Sepehrkhouy, S.; Meijer, M.; de Cock, H.; Haas, P.J.

    2013-01-01

    Background: Aspergillus tubingensis is a black Aspergillus belonging to the Aspergillus section Nigri, which includes species that morphologically resemble Aspergillus niger. Recent developments in species determination have resulted in clinical isolates presumed to be Aspergillus niger being

  7. Xylanase production by a local fungal isolate, Aspergillus niger USM AI 1 via solid state

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    Ibrahim Che Omar

    2005-03-01

    Full Text Available Isolate USM A1 I which was identified to be Aspergillus niger was selected as a potential producer of xylanase via a solid state fermentation system (SSF using palm kernel cake (PKC as substrate. The modification of the physical conditions of the SSF system indicated that the xylanase activity was 23.97 U/g PKC at the moisture ratio of 1:0.75 of PKC: moistening agent with the inoculum size of 1¥104 spores/ml and cultivated at the ambient temperature (28±3ºC. The supplementation of additional carbon and nitrogen sources in the PKC medium could enhance enzyme productivity. The maximum production of xylanase and growth obtained with the supplementation of xylose at 0.75% (w/w were 25.40 U/g and 1.69 mg glucosamine/ g PKC. Moreover, the presence of NaNO3 at 0.075% (w/w as additional nitrogen source further enhanced xylanase production to 33.99 U/g PKC although the growth remained unchanged at about 1.67 mg glucosa- mine/g PKC. The optimized conditions showed an increased xylanase production by 157% compared to before the optimization of the SSF system. The xylanase productivity was 23.12 U/mg glucosamine after optimization and 11.72 U/mg glucosamine before optimization.

  8. Bioleaching of nickel by Aspergillus humicola SKP102 isolated from Indian lateritic overburden

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    Suchhanda Ghosh

    2016-01-01

    Full Text Available The lateritic deposits spread over the Eastern Ghats of Sukinda Valley, Odisha, India, produce a huge amount of overburden annually as a byproduct of chromite mining. This chromite mining overburden contains nickel, the only source of the metal in the country. During this study Aspergillus humicola SKP102, an indigenous fungus isolated from the mining overburden was employed for the leaching of nickel. About 53.89% of the nickel could be leached by the fungus when grown in batch mode using a Czapek dox medium containing 2% (w/v of the mining overburden. The parameters affecting bioleaching were optimized in order to grow the fungus and leach the metal. Of the different options of cheap carbon sources, straw infusion and molasses emerged as viable options for the growth of the fungus and the leaching of nickel. Two-step and indirect techniques were also used for this purpose, and they resulted in 53.09% and 65.04% Ni leaching respectively. Adding diluted sulfuric acid to the leaching medium resulted in 97.05% nickel recovery from the overburden pulp. A. humicola SKP102 could be a potential tool for leaching nickel from the mining overburden.

  9. Reproductive isolation and the expansion of an invasive hybrid swarm

    Science.gov (United States)

    Blum, Michael J.; Walters, David M.; Burkhead, Noel M.; Freeman, Byron J.; Porter, Brady A.

    2010-01-01

    Biological invasions involving hybridization proceed according to prezygotic and postzygotic reproductive isolating mechanisms. Yet few comparisons of reproductive isolation have been carried out to understand how different mechanisms prevent or promote invasions involving hybridization. Here we present a study of prezygotic and postzygotic isolation between non-native red shiner (Cyprinella lutrensis) and native blacktail shiner (C. venusta stigmatura) from the Coosa River basin (USA) to better understand the formation and expansion of invasive hybrid swarms. We conducted spawning trials to measure mating preferences and raised broods from crosses to assay hybrid viability through early juvenile development. Females of both species were more responsive to conspecific mates, although blacktail shiner females responded more often to heterospecific mates than did red shiner females. Fecundity of red shiner females was also higher than blacktail shiner females. Heterospecific crosses resulted in lower fertilization and egg hatching rates, but we found no other evidence of inviability. Rather, we found comparatively low larval mortality of F1 hybrids, which is suggestive of heterosis. These findings support prior inferences of assortative mating from genetic descriptions of hybridization, and that the invasion in the Coosa River is likely proceeding due to interspecific competition and intrinsic hybrid viability.

  10. Isolation, identification of an axenic fungal isolate of aspergillus sp. (mbl-1511) and its subsequent improvement for enhanced extracellular lipolytic potential through monoculture fermentation

    International Nuclear Information System (INIS)

    Iftikhar, T.; Sidra, A.; Ali, M.; Majeed, H.; Abdullah, R.

    2017-01-01

    The present investigation was conducted for extracellular lipases production. One hundred and forty samples of fungi were isolated from different environment and food samples. Among all the isolated cultures, an isolate obtained from chicken roasted in oil (MBL-1511) gave the highest extracellular lipase through SSF. Hyper producer strain (MBL-1511) was morphologically identified. A morphologically identified isolate of Aspergillus niger (MBL 1511) was verified by DNA barcoding marker like 18S rRNA gene sequence. The sequence of Aspergillus niger (MBL 1511) was registered with accession no. [GenBank: KP172477] in the public nucleotide library (genbank) of NCBI. The selected hyper producer of Aspergillus niger (MBL-1511) strain was subjected to physical and chemical mutagenic treatments to improve its lipolytic potential. Proximate analysis confirmed brassica meal as the best basal substrate with the lipases potential of 10.67+-0.01 IU/mL (wild) and 19.58+-0.04 IU/mL (mutant). The optimum conditions for the maximized extracellular lipases production were 1.0 mL inoculum at 30 degree C after 72 h at pH of 6.2. Finally, a potent mutant of A. niger [MBL-1511SA-4(150 min)] with an increased activity of 161 % over the wild strain was obtained when olive oil was used at 1% (v/v) concentration. (author)

  11. In vitro synergy of natamycin and voriconazole against clinical isolates of Fusarium, Candida, Aspergillus and Curvularia spp.

    Science.gov (United States)

    Sradhanjali, Swatishree; Yein, Bandana; Sharma, Savitri; Das, Sujata

    2018-01-01

    To determine the minimum inhibitory concentrations (MICs) of voriconazole and natamycin, alone and in combination, against the clinical isolates of Fungus and to evaluate the synergy between the drugs in an experimental in vitro study. In an experimental in vitro study, clinical isolates of Fusarium , Aspergillus , Candida and Curvularia spp were maintained on Sabouraud Dextrose Agar and used for the study. The MICs of natamycin and voriconazole, used alone and in combination, were evaluated by checkerboard microdilution technique based on the standard protocol proposed by the Clinical Laboratory Standards Institute. The interactions were assessed using the fractional inhibitory concentration (FIC) Index model. Tested with all the clinical isolates, the MICs ranged between 0.125 and 8 µg/mL both for natamycin and voriconazole. In descending order, maximum synergism (FIC ≤0.5) was observed in Candida spp (33.3%) followed by Curvularia spp and Fusarium spp (23.1%). Synergism was least for Aspergillus spp (22.2%). However, at 61.5% (8/13), maximum additive effect (>0.5-1) was observed in Aspergillus spp and minimum (33.3%, 2/6) in Candida spp. Indifference (FIC value >1 and≤4) was observed in 22.2% (2/9) of Aspergillus spp, 15.4% (2/13) of Fusarium spp, 33.3% (2/6) of Candida spp and 23.1% (3/13) of Curvularia spp. No cases of antagonism (FIC >4) were observed. Natamycin and voriconazole in combination demonstrated more effective antifungal activity than single-use in vitro treatment in all species tested, which implies that these combinations may be helpful in treating fungal keratitis. There was no antagonism between these two drugs. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

  12. New bisabolane sesquiterpenoids from a marine-derived fungus Aspergillus sp. isolated from the sponge Xestospongia testudinaria.

    Science.gov (United States)

    Sun, Ling-Ling; Shao, Chang-Lun; Chen, Jian-Feng; Guo, Zhi-Yong; Fu, Xiu-Mei; Chen, Min; Chen, Yi-Yan; Li, Rui; de Voogd, Nicole J; She, Zhi-Gang; Lin, Yong-Cheng; Wang, Chang-Yun

    2012-02-01

    Three new phenolic bisabolane sesquiterpenoid dimers, disydonols A-C (1-3), and one known compound (S)-(+)-sydonol (4) were isolated from the fermentation broth of a marine-derived fungus Aspergillus sp., which was isolated from the sponge Xestospongia testudinaria collected from the South China Sea. Their structures were elucidated on the basis of comprehensive spectral analysis including 1D and 2D NMR spectra and HR-ESI-MS. These compounds were evaluated for cytotoxic activity against HepG-2 and Caski human tumour cell lines. Among them, compounds 1 and 3 exhibited cytotoxicity against the two cell lines. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Occurrence of fungi and cytotoxicity of the species: Aspergillus ochraceus, Aspergillus niger and Aspergillus flavus isolated from the air of hospital wards.

    Science.gov (United States)

    Gniadek, Agnieszka; Krzyściak, Paweł; Twarużek, Magdalena; Macura, Anna B

    2017-03-30

    The basic care requirement for patients with weakened immune systems is to create the environment where the risk of mycosis is reduced to a minimum. Between 2007 and 2013 air samples were collected from various wards of a number of hospitals in Kraków, Poland, by means of the collision method using MAS-100 Iso MH Microbial Air Sampler (Merck Millipore, Germany). The air mycobiota contained several species of fungi, and almost 1/3 of it was made up of the species of the Aspergillus genus. Sixty-one strains of species other than A. fumigatus were selected for the research purposes, namely: 28 strains of A. ochraceus, 22 strains of A. niger and 11 strains of A. flavus species. Selected fungi underwent a cytotoxicity evaluation with the application of the MTT colorimetric assay (3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide). The assay assesses cell viability by means of reducing the yellow tetrazolium salt to insoluble formazan. A semi-quantitative scale for cytotoxicity grading was adopted: low cytotoxic effect (+) with half maximal inhibitory concentration (IC50) for values ranging from 31.251 cm2/ml to 7.813 cm2/ml, medium cytotoxic effect (++) for values ranging from 3.906 cm2/ml to 0.977 cm2/ml and the high one (+++) for values ranging from 0.488 cm2/ml to 0.061 cm2/ml. The absence of cytotoxicity was determined when the IC50 values was at ≥ 50. For 48 samples the analyzed fungi displayed the cytotoxic effect with A. ochraceus in 26 out of 28 cases, with 11 strains displaying the high cytotoxic effect. The lowest cytotoxicity was displayed by fungi of A. niger in 13 out of 22 cases, and the major fungi of A. flavus species were toxic (9 out of 11 cases). A half of the fungi displayed the low cytotoxic effect. On the basis of the comparison of average cytotoxicity levels it was determined that there were significant differences in the levels of cytotoxicity of the analyzed fungi. However, such statement may not provide grounds for a definite

  14. Local isolate of Saccharomyces cerevisiae as biocompetitive agent of Aspergillus flavus

    Directory of Open Access Journals (Sweden)

    Eni Kusumaningtyas

    2006-12-01

    Full Text Available Aspergillus flavus is a toxigenic fungus that contaminates feed and influences the animal health. Saccharomyces cerevisiae can be used as a biocompetitive agent to control the contamination. The ability of local isolate of S. cerevisiae as a biocompetitive agent for A. flavus was evaluated. A. flavus (30ml was swept on Sabouraud dextrose agar (SDA, while S. cerevisiae was swept on its left and right. Plates were incubated at 28oC for nine days. Lytic activity of S. cerevisiae was detected by pouring its suspension on the centre of the cross streaks of A. flavus. Plates were incubated at 28oC for five days. Growth inhibition of A. flavus by S. cerevisiae was determined by mixing the two fungi on Potato dextrose broth and incubated at 28oC for 24 hours. Total colony of A. flavus were then observed at incubation time of 2, 4, 6 and 24 hours by pour plates method on the SDA plates and incubated on 28oC for two days. Growth of hyphae of A. flavus sweep were inhibited with the swept of S. cerevisiae. The width of A. flavus colony treated with S. cerevisiae is narrower (3,02 cm than that of control ( 4,60 cm. The growth of A. flavus was also inhibited on the centre of cross streak where the S. cerevisiae poured. S. cerevisiae gradually reduced the colony number of A. flavus in the mixed culture of broth fungi ie. 14 x 103 CFU/ml while colony number of control is 80 x 103 CFU/ml. Results showed that S. cerevisiae could be used as biocompetitive agent of A. flavus.

  15. Molecular Characterization and Antimicrobial Activity of an Endolichenic Fungus, Aspergillus sp. Isolated from Parmelia caperata of Similipal Biosphere Reserve, India.

    Science.gov (United States)

    Padhi, Srichandan; Das, Devaranjan; Panja, Suraj; Tayung, Kumananda

    2017-06-01

    Endolichenic fungi are microbes that inhabit healthy inner lichen tissues without any disease symptoms. They have been reported to produce new and interesting bioactive metabolites. In the present study, an endolichenic fungus frequently isolated from surface-sterilized lichen thallus of Parmelia caperata has been described. The fungus was identified as Aspergillus tubingensis based on morphological traits and ITS rDNA sequence. Crude metabolites extracted from the culture broth exhibited considerable antimicrobial activity against a panel of clinically significant human pathogens. The fungus showed optimum antimicrobial activity in PDB medium in day 7 of incubation period. PDB medium amended with 1 % NaCl and at alkaline pH was found to be optimal for antimicrobial metabolites production. Enhanced activity was observed when the fungus was exposed briefly to a heat shock of 60 °C during incubation. The metabolites showed optimum λ-max at 214 nm with an absorbance value of 1.589. Molecular characterization of the isolate was carried out by ITS phylogeny and ITS2 secondary structure analyses. The phylogenetic trees based on both ITS rDNA and ITS2 sequences showed the isolate within the clade A. tubingensis. Considering the ubiquity and ambiguity in identifying Aspergillus species of different lifestyles, a method to differentiate pathogenic and endophytic Aspergillus at species level was developed using ITS2 secondary structure analysis. The results showed common folding pattern in the secondary structures with a helix and a 5' dangling end found to be highly conserved. Certain features in the secondary structure like multi-bulges and a symmetric interior loop were observed to be unique which distinguish our isolate from other A. tubingensis.

  16. Mycotoxin production and predictive modelling kinetics on the growth of Aspergillus flavus and Aspergillus parasiticus isolates in whole black peppercorns (Piper nigrum L).

    Science.gov (United States)

    Yogendrarajah, Pratheeba; Vermeulen, An; Jacxsens, Liesbeth; Mavromichali, Evangelia; De Saeger, Sarah; De Meulenaer, Bruno; Devlieghere, Frank

    2016-07-02

    The growth and mycotoxin production of three Aspergillus flavus isolates and an Aspergillus parasiticus isolate were studied in whole black peppercorns (Piper nigrum L.) using a full factorial design with seven water activity (aw) (0.826-0.984) levels and three temperatures (22, 30 and 37°C). Growth rates and lag phases were estimated using linear regression. Diverse secondary models were assessed for their ability to describe the radial growth rate as a function of individual and combined effect of aw and temperature. Optimum radial growth rate ranged from 0.75±0.04 to 2.65±0.02mm/day for A. flavus and 1.77±0.10 to 2.50±0.10mm/day for A. parasiticus based on the Rosso cardinal estimations. Despite the growth failure of some isolates at marginal conditions, all the studied models showed good performance to predict the growth rates. Validation of the models was performed on independently derived data. The bias factors (0.73-1.03), accuracy factors (0.97-1.36) and root mean square error (0.050-0.278) show that the examined models are conservative predictors of the colony growth rate of both fungal species in black peppers. The Rosso cardinal model can be recommended to describe the individual aw effect while the extended Gibson model was the best model for describing the combined effect of aw and temperature on the growth rate of both fungal species in peppercorns. Temperature optimum ranged from 30 to 33°C, while aw optimum was 0.87-0.92 as estimated by multi-factorial cardinal model for both species. The estimated minimum temperature and aw for A. flavus and A. parasiticus for growth were 11-16°C and 0.73-0.76, respectively, hence, achieving these conditions should be considered during storage to prevent the growth of these mycotoxigenic fungal species in black peppercorns. Following the growth study, production of mycotoxins (aflatoxins B1, B2, G1, G2, sterigmatocystin and O-methyl sterigmatocystin (OMST)) was quantified using LC-MS/MS. Very small

  17. Isolation of culturable mycobiota from agricultural soils and determination of tolerance to glyphosate of nontoxigenic Aspergillus section Flavi strains.

    Science.gov (United States)

    Carranza, Cecilia S; Barberis, Carla L; Chiacchiera, Stella M; Dalcero, Ana María; Magnoli, Carina E

    2016-01-01

    Glyphosate-based herbicides are extensively used in Argentina's agricultural system to control undesirable weeds. This study was conducted to evaluate the culturable mycobiota [colony forming units (CFU) g(-1) and frequency of fungal genera or species] from an agricultural field exposed to pesticides. In addition, we evaluated the tolerance of A. oryzae and nontoxigenic A. flavus strains to high concentrations (100 to 500 mM - 17,000 to 84,500 ppm) of a glyphosate commercial formulation. The analysis of the mycobiota showed that the frequency of the main fungal genera varied according to the analyzed sampling period. Aspergillus spp. or Aspergillus section Flavi strains were isolated from 20 to 100% of the soil samples. Sterilia spp. were also observed throughout the sampling (50 to 100%). Aspergillus section Flavi tolerance assays showed that all of the tested strains were able to develop at the highest glyphosate concentration tested regardless of the water availability conditions. In general, significant reductions in growth rates were observed with increasing concentrations of the herbicide. However, a complete inhibition of fungal growth was not observed with the concentrations assayed. This study contributes to the knowledge of culturable mycobiota from agricultural soils exposed to pesticides and provides evidence on the effective growth ability of A. oryzae and nontoxigenic A. flavus strains exposed to high glyphosate concentrations in vitro.

  18. A Simple Procedure to Evaluate Competitiveness of Toxigenic and Atoxigenic Isolates of Aspergillus flavus in Solid and Liquid Media

    Directory of Open Access Journals (Sweden)

    Mohammad Moradi

    2017-04-01

    Full Text Available Application of atoxigenic strains of Aspergillus flavus to reduce aflatoxin levels is the most successful strategy applied in some agricultural crops. The role of ammonium hydroxide for preliminary screening of the competitiveness of atoxigenic A. flavus isolates to interfere with aflatoxin production by highly toxigenic isolates were evaluated. Out of 270 A. flavus isolates, 17 were detected as true atoxigenic using cultural methods and confirmed by analytical assays from different pistachio agro-ecological zones during 2013. For assessment competitive ability among atoxigenic isolates of A. flavus with highly toxigenic one, rice flour, coconut agar and coconut broth medium substrates were inoculated with mixtures including combinations of toxigenic and atoxigenic isolates, simultaneously. The rice flour substrate was used to quantify the content of aflatoxin in either co-inoculations or toxigenic isolate alone on thin layer chromatography plates with a scanning densitometer. While the culture media were used to determine the intensity of color change on exposing to ammonium hydroxide vapor. The reduction rates of aflatoxin B1 in co-inoculations were varied and ranged from 2%-82%. Based on the intensity of colony color changes, the competitiveness of the isolates was classified into five groups. Atoxigenic isolates with high competitiveness have shown low color changes in culture media and high aflatoxin reduction in TLC assays with a ratio of higher than 78%. The method will facilitate preliminary screening of efficient atoxigenic isolates for mitigation of aflatoxins in food and feed as a cheap, simple and quick method.

  19. Characterization and biological treatment of colored textile wastewaters from the typical Tunisian hat Chechia dyeing using newly isolated Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Hajer Barouni

    2016-09-01

    Full Text Available This study aimed to characterize and investigate, for the first time, the treatment of real colored wastewaters from the artisanal dyeing of the typical Tunisian hat Chechia, using a newly isolated fungal strain. This textile effluent was a mixture called Mix of colored wastewaters from the three main types of Chechia. The major pollutant of the Mix was the toxic Azo dye Amaranth Acid or Acid Red 27. The fungal strain that made the cleanup was discovered in a Chechia dyeing wastewater’s container and identified by ITS rDNA gene sequencing. This isolated Aspergillus niger showed interesting performances on the demonstration of Chechia wastewater’s biodegradation in batch cultures. In order to understand the effect of agitation, Mix dilution and inoculum size on decolourisation and pollution removal, a full factorial experimental design 23 was set up. At the optimal conditions which were 20% inoculum size, 25% Chechia Mix dilution and an agitation of 100 rpm, Aspergillus niger was able to remove color as high as 70.18±2.84% at an initial dye concentration of 1346.6±0.01 mg/L, and to reduce COD to 74.17±14.52% at an initial COD of 4157±422 mg/L. FT-IR spectra analysis confirmed the decolourisation by biodegradation and transformation of the dyes. The treatment by the isolated Aspergillus niger could be successfully applied as a sustainable method to solve one of handicraft dyeing plants environmental management issues.

  20. Isolation and NMR Characterization of Fumonisin B-2 and a New Fumonisin B-6 from Aspergillus niger

    DEFF Research Database (Denmark)

    Månsson, Maria; Klejnstrup, Marie Louise; Phipps, Richard Kerry

    2010-01-01

    A new fumonisin, fumonisin B-6 (1), has been isolated by cation-exchange and reverse-phase chromatography, together with fumonisin B-2 (2), from,stationary cultures of the fungus Aspergillus niger NRRL 326. Analysis of mass spectrometric and NMR data determined that FB6 is a positional isomer...... of FBI and iso-FB1, having hydroxyl functions at C3, C4, and C5. Analysis of the NMR data for FB2 showed very similar chemical shift values when compared to an authentic Fusarium FB2 standard, strongly indicating identical molecules despite that an absolute stereochemical assignment of FB2 from A. niger...

  1. Variability among strains of Aspergillus section Nigri with capacity to degrade tannic acid isolated from extreme environments.

    Science.gov (United States)

    Lara-Victoriano, F; Veana, F; Hernández-Castillo, F D; Aguilar, C N; Reyes-Valdés, M H; Rodríguez-Herrera, R

    2017-01-01

    Tannins are polyphenolic compounds that cause astringent flavor and turbidity in food. Tannase is an enzyme that catalyzes the hydrolysis of tannins and is used in food industry. This study was conducted to determine the genetic variability and the tannase alleles variation in fungal strains isolated from soil and plants at five extreme areas of Coahuila, México. Two screening assays under 1 and 20 % of tannic acid were performed, with the isolations. In these assays, it was possible to identify 756 and 128 fungal strains, respectively. The major fungal variability was observed in "Cuatro Ciénegas" with 26 strains. The microorganisms were distributed in 11 groups, which correspond to Aspergillus section Nigri. AN7 and AN1 groups showed the major number of isolates from "Paila" and "Cuatro Ciénegas" locations, respectively. In the last location, the major diversity and specific richness were found. But in "Ojo Caliente," tannase allele conservations were observed.

  2. Effect of Essential Oils of Syzygium aromaticum and Melaleuca alternifolia on Isolates of Aspergillus sp.

    Directory of Open Access Journals (Sweden)

    A. Lima

    2014-03-01

    Full Text Available Essential  oils  have  antimicrobial  substances,  lower  cost  and  the  lower  resistance  of microorganism. This study aimed to evaluate the inhibition of mycelial growth of Aspergillus flavus and  A.  niger  with  essential  oils  of  Syzygium  aromaticum  and  Melaleuca  alternifolia.  For  this, aliquots  (0.5,  5,  10  and  15μL  of  essential  oils  from  S.  aromaticum  and  M.  alternifolia   were distributed on the surface of the culture medium Potato Dextrose Agar (PDA with Drigaslsky. In the control treatment were used only plates containing PDA plus chloramphenicol (1%. After 2 hours, a disc (8mm diameter of the isolated mycelium, with 10 days of age on PDA, was peaked to the center of the plates, these plates were sealed and incubated at 282°C, in the dark. The essential  oil  of  S.  aromaticum  inhibited  the  mycelial  growth  of  A.  flavus  and  A.  niger,  in  all aliquots. When using the essential oil of M. alternifolia decreased the mycelial growth of A. flavus in all aliquots, however, showed a low efficiency in control of A. niger. It was concluded that the essential  oil  S.  aromaticum  is  effective  against  A.  flavus  and  A.  niger,  which  can  be  used  in control against these microorganisms, and M. alternifolia not show satisfactory results in relation to reducing the growth of pathogens evaluated.

  3. Gene expression profiles of Aspergillus flavus isolates responding to oxidative stress in different culture media

    Science.gov (United States)

    Aflatoxin contamination of peanut by Aspergillus flavus is exacerbated by drought stress. Drought also stimulates the production of reactive oxygen species (ROS) in plant tissues implying a correlation between ROS and aflatoxin production. Here, we performed gene expression analysis by RNAseq of tox...

  4. Comparative transcriptome analysis of Aspergillus flavus isolates under different oxidative stresses and culture media

    Science.gov (United States)

    Aspergillus flavus and aflatoxin contamination in the field are known to be influenced by numerous stress factors, particularly drought and heat stress. However, the purpose of aflatoxin production is unknown. Here, we report transcriptome analyses comprised of 282.6 Gb of sequencing data describing...

  5. Characterization of Aspergillus section Flavi isolated from organic Brazil nuts using a polyphasic approach.

    Science.gov (United States)

    Reis, T A; Baquião, A C; Atayde, D D; Grabarz, F; Corrêa, B

    2014-09-01

    Brazil nut (Bertholletia excelsa), an important non-timber forest product from Amazonia, is commercialized in worldwide markets. The main importers of this nut are North America and European countries, where the demand for organic products has grown to meet consumers concerned about food safety. Thus, the precise identification of toxigenic fungi is important because the Brazil nut is susceptible to colonization by these microorganisms. The present study aimed to characterize by polyphasic approach strains of Aspergillus section Flavi from organic Brazil nuts. The results showed Aspergillus flavus as the main species found (74.4%), followed by Aspergillus nomius (12.7%). The potential mycotoxigenic revealed that 80.0% of A. flavus were toxin producers, 14.3% of which produced only aflatoxin B (AFB), 22.85% of which produced only cyclopiazonic acid (CPA), and 42.85% produced both them. All strains of A. nomius were AFB and AFG producers and did not produce CPA. There is no consensus about what Aspergillus species predominates on Brazil nuts. Apparently, the origin, processing, transport and storage conditions of this commodity influence the species that are found. The understanding about population of fungi is essential for the development of viable strategies to control aflatoxins in organic Brazil nuts. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Isolation and Characterization of the mheA (Most Highly Expressed) Gene of Aspergillus oryzae

    Science.gov (United States)

    Liu, Pengcheng; Lim, Ji-Young; Kim, Hee-Seo; Kim, Jong Hwa

    2012-01-01

    The amino acid sequence of the mheA gene of Aspergillus oryzae encodes a putative metallothionein-like protein 1. The size of the mheA transcript was 497 nt and the mheA promoter was induced by glucose, consistent with results of analysis by Northern hybridization and with the pdcA promoter, respectively. PMID:23115516

  7. Seed isolates of Alternaria and Aspergillus fungi increase germination of Astragalus utahensis

    Science.gov (United States)

    Sean D. Eldredge; Brad Geary; Scott L. Jensen

    2016-01-01

    Astragalus utahensis (Torr.) Torr. & A. Gray (Fabaceae) (Utah milkvetch) is native lo the arid Great Basin and has desirable attributes that make it a good candidate for restoration in arid, noncompetitive situations. Seed dormancy is a significant barrier to consistent establishment for this species. Species of Alternaria and Aspergillus fungi have...

  8. Morphological Characterization and Determination of Aflatoxin-Production Potentials of Aspergillus flavus Isolated from Maize and Soil in Kenya

    Directory of Open Access Journals (Sweden)

    Matome Gabriel Thathana

    2017-09-01

    Full Text Available This study aimed at morphologically identifying Aspergillus flavus in soil and maize and at determining their aflatoxin-producing potentials. Five hundred and fourteen isolates obtained from maize and soil in Kenya were cultivated on Czapeck Dox Agar, Malt Extract Agar, Sabouraud Dextrose Agar, Potato Dextrose Agar, and Rose-Bengal Chloramphenicol Agar. Isolates were identified using macro-morphological characteristics. Micromorphological characteristics were determined using slide cultures. Aflatoxin production was determined by direct visual determination of the UV fluorescence of colonies on Coconut Agar Medium, Yeast Extract Sucrose agar, and Yeast Extract Cyclodextrin Sodium Deoxycholate agar and by Thin Layer Chromatography. Forty-three presumptive A. flavus isolates were identified; aflatoxin was detected in 23% of the isolates by UV fluorescence screening and in 30% by Thin-Layer Chromatography (TLC. The aflatoxins produced were: aflatoxin B1 (AFB1, aflatoxin B2 (AFB2, and aflatoxin G1 (AFG1; some isolates produced only AFB1, whereas others produced either AFB1 and AFB2 or AFB1 and AFG1. The highest incidence of A. flavus (63% and aflatoxin production (28% was recorded in samples from Makueni District. Isolates from Uasin Gishu (21% and Nyeri (5% were non-aflatoxigenic. Bungoma District recorded 11% positive isolates of which 2% were aflatoxin producers. The occurrence of aflatoxin-producing A. flavus emphasises the need for measures to eliminate their presence in food crops.

  9. Isolation of Aspergillus fumigatus from sputum is associated with elevated airborne levels in homes of patients with asthma.

    Science.gov (United States)

    Fairs, A; Agbetile, J; Bourne, M; Hargadon, B; Monteiro, W R; Morley, J P; Edwards, R E; Wardlaw, A J; Pashley, C H

    2013-08-01

    Indoor bioaerosols, such as mold spores, have been associated with respiratory symptoms in patients with asthma; however, dose-response relationships and guidelines on acceptable levels are lacking. Furthermore, a causal link between mold exposure and respiratory infections or asthma remains to be established. The aim of this study was to determine indoor concentrations of Aspergillus fumigatus and a subset of clinically relevant fungi in homes of people with asthma, in relation to markers of airways colonization and sensitization. Air and dust samples were collected from the living room of 58 properties. Fungal concentrations were quantified using mold-specific quantitative PCR and compared with traditional microscopic analysis of air samples. Isolation of A. fumigatus from sputum was associated with higher airborne concentrations of the fungus in patient homes (P = 0.04), and a similar trend was shown with Aspergillus/Penicillium-type concentrations analyzed by microscopy (P = 0.058). No association was found between airborne levels of A. fumigatus and sensitization to this fungus, or dustborne levels of A. fumigatus and either isolation from sputum or sensitization. The results of this study suggest that the home environment should be considered as a potential source of fungal exposure, and elevated home levels may predispose people with asthma to airways colonization. © 2012 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  10. Entomopathogens Isolated from Invasive Ants and Tests of Their Pathogenicity

    Directory of Open Access Journals (Sweden)

    Maria Fernanda Miori de Zarzuela

    2012-01-01

    Full Text Available Some ant species cause severe ecological and health impact in urban areas. Many attempts have been tested to control such species, although they do not always succeed. Biological control is an alternative to chemical control and has gained great prominence in research, and fungi and nematodes are among the successful organisms controlling insects. This study aimed to clarify some questions regarding the biological control of ants. Invasive ant species in Brazil had their nests evaluated for the presence of entomopathogens. Isolated entomopathogens were later applied in colonies of Monomorium floricola under laboratory conditions to evaluate their effectiveness and the behavior of the ant colonies after treatment. The entomopathogenic nematodes Heterorhabditis sp. and Steinernema sp. and the fungi Beauveria bassiana, Metarhizium anisopliae, and Paecilomyces sp. were isolated from the invasive ant nests. M. floricola colonies treated with Steinernema sp. and Heterorhabditis sp. showed a higher mortality of workers than control. The fungus Beauveria bassiana caused higher mortality of M. floricola workers. However, no colony reduction or elimination was observed in any treatment. The defensive behaviors of ants, such as grooming behavior and colony budding, must be considered when using fungi and nematodes for biological control of ants.

  11. Isolation and characterization of a beta-primeverosidase-like endo-manner beta-glycosidase from Aspergillus fumigatus AP-20.

    Science.gov (United States)

    Yamamoto, Shigeru; Okada, Masamichi; Usui, Taichi; Sakata, Kanzo

    2002-04-01

    A novel beta-glycosidase-producing microorganism was isolated from soil and identified as Aspergillus fumigatus AP-20 based on its taxonomical characteristics. The enzyme was found to be an extracellular protein in the culture of the isolated fungus and was purified 88-fold by fractionation with ammonium sulfate followed by successive column chromatographies on phenyl-Sepharose HP and Mono P HR. The molecular mass was estimated to be 47 kDa by SDS-PAGE and the isoelectric point to be pH 6.0 by isoelectric focusing. The purified enzyme was highly specific for a substrate, p-nitrophenyl beta-primeveroside (6-O-beta-D-xylopyranosyl-beta-D-glucopyranoside), which was cleaved in an endo-manner into primeverose and p-nitrophenol.

  12. Clinical Isolates of Aspergillus Species Remain Fully Susceptible to Voriconazole in the Post-Voriconazole Era▿

    OpenAIRE

    Guinea, Jesús; Recio, Sandra; Peláez, Teresa; Torres-Narbona, Marta; Bouza, Emilio

    2008-01-01

    We studied the activity of voriconazole against 400 clinical strains of Aspergillus from the pre-voriconazole (1999 to 2002) and post-voriconazole (2003 to 2007) periods. Although the mean MICs of strains from the post-voriconazole period were slightly higher (0.39 versus 0.57 μg/ml; P < 0.001), all strains were susceptible to voriconazole and presented an MIC of ≤2 μg/ml.

  13. Plasmodium falciparum field isolates from South America use an atypical red blood cell invasion pathway associated with invasion ligand polymorphisms.

    Directory of Open Access Journals (Sweden)

    Mary Lopez-Perez

    Full Text Available Studies of Plasmodium falciparum invasion pathways in field isolates have been limited. Red blood cell (RBC invasion is a complex process involving two invasion protein families; Erythrocyte Binding-Like (EBL and the Reticulocyte Binding-Like (PfRh proteins, which are polymorphic and not fully characterized in field isolates. To determine the various P. falciparum invasion pathways used by parasite isolates from South America, we studied the invasion phenotypes in three regions: Colombia, Peru and Brazil. Additionally, polymorphisms in three members of the EBL (EBA-181, EBA-175 and EBL-1 and five members of the PfRh (PfRh1, PfRh2a, PfRh2b, PfRh4, PfRh5 families were determined. We found that most P. falciparum field isolates from Colombia and Peru invade RBCs through an atypical invasion pathway phenotypically characterized as resistant to all enzyme treatments (NrTrCr. Moreover, the invasion pathways and the ligand polymorphisms differed substantially among the Colombian and Brazilian isolates while the Peruvian isolates represent an amalgam of those present in the Colombian and Brazilian field isolates. The NrTrCr invasion profile was associated with the presence of the PfRh2a pepC variant, the PfRh5 variant 1 and EBA-181 RVNKN variant. The ebl and Pfrh expression levels in a field isolate displaying the NrTrCr profile also pointed to PfRh2a, PfRh5 and EBA-181 as being possibly the major players in this invasion pathway. Notably, our studies demonstrate the uniqueness of the Peruvian P. falciparum field isolates in terms of their invasion profiles and ligand polymorphisms, and present a unique opportunity for studying the ability of P. falciparum parasites to expand their invasion repertoire after being reintroduced to human populations. The present study is directly relevant to asexual blood stage vaccine design focused on invasion pathway proteins, suggesting that regional invasion variants and global geographical variation are likely to

  14. Comparison of Two Molecular Assays for Detection and Characterization of Aspergillus fumigatus Triazole Resistance and Cyp51A Mutations in Clinical Isolates and Primary Clinical Samples of Immunocompromised Patients

    Directory of Open Access Journals (Sweden)

    Patricia Postina

    2018-03-01

    Full Text Available In hematological patients, the incidence of invasive aspergillosis (IA caused by azole resistant Aspergillus fumigatus (ARAf is rising. As the diagnosis of IA is rarely based on positive culture in this group of patients, molecular detection of resistance mutations directly from clinical samples is crucial. In addition to the in-house azole resistance ARAf polymerase chain reaction (PCR assays detecting the frequent mutation combinations TR34/L98H, TR46/Y121F/T289A, and M220 in the Aspergillus fumigatus (A. fumigatus Cyp51A gene by subsequent DNA sequence analysis, we investigated in parallel the commercially available AsperGenius® real time PCR system in detecting the Cyp51A alterations TR34/L98H and Y121F/T289A directly from 52 clinical samples (15 biopsies, 22 bronchoalveolar lavage (BAL, 15 cerebrospinal fluid (CSF samples and ARAf isolates (n = 3 of immunocompromised patients. We analyzed DNA aliquots and compared both methods concerning amplification and detection of Aspergillus DNA and Cyp51A alterations. As positive control for the feasibility of our novel Y121F and T289A PCR assays, we used two A. fumigatus isolates with the TR46/Y121F/T289A mutation combination isolated from hematological patients with known Cyp51A alterations and a lung biopsy sample of a patient with acute myeloid leukemia (AML. The rate of positive ARAf PCR results plus successful sequencing using the ARAf PCR assays was 61% in biopsies, 29% in CSF, 67% in BAL samples and 100% in isolates. In comparison the amount of positive PCRs using the AsperGenius® assays was 47% in biopsies, 42% in CSF, 59% in BAL samples and 100% in isolates. Altogether 17 Cyp51A alterations were detected using our ARAf PCRs plus DNA sequencing and therefrom 10 alterations also by the AsperGenius® system. The comparative evaluation of our data revealed that our conventional PCR assays are more sensitive in detecting ARAf in BAL and biopsy samples, whereby differences were not significant

  15. Molecular and biochemical characterization of soil isolates of Aspergillus niger aggregate and an assessment of their antagonism against Rhizoctonia solani

    Directory of Open Access Journals (Sweden)

    M.R. Khan

    2007-12-01

    Full Text Available Sixteen Aspergillus niger aggregate isolates collected from different crop fi elds were subjected to RAPDPCR using 20 Operon primers and 8 synthetic primers. Twenty-two primers led to the amplifi cation of 727 fragments ranging from 3500 bp (OPA 11 to 200 bp (primer 06. Two bands were monomorphic, while the rest were polymorphic. Three amplicons produced by OPA 16 were recorded as isolate-specifi c as 2300 bp by AnC2 and AnR3, and as 2800 bp by AnC2 only. The highest genetic similarity (0.79 was measured between AnC2 and AnR3, and the lowest (0.17 between AnC2 and AnR2. Multivariate analysis of genetic similarity revealed three major clusters, named group I, group II and group III. All isolates were ochratoxin A negative (<1 ng g-1. Isolates AnC2 and AnR3, which produced HCN and solubilized the greatest amount of phosphorus, belonged to group I. These isolates also signifi cantly increased eggplant yield and caused the greatest inhibition of colonization by R. solani in dual culture. They also suppressed the root rot on eggplant and the soil population of R. solani in pot soil.

  16. Isolation and optimization of pectinase enzyme production one of useful industrial enzyme in Aspergillus niger, Rhizopus oryzae, Penicilium chrysogenum

    Directory of Open Access Journals (Sweden)

    akram songol

    2016-06-01

    Full Text Available Introduction: Pectinase enzyme is one of the most important industrial enzymes which isolated from a wide variety of microorganisms such as bacteria and filamentous fungi. This enzyme has been usually used in the fruit and textile industry. In this study, the isolation and optimization of pectinase-producing fungi on decaying rotten fruits were studied. Materials and methods: Isolation and screening of pectinase producing fungi performed through plate culture on pectin medium and staining with Lugol's iodine solution. The best strains were identified by ITS1, 4 sequencing as Aspergillus fumigatus, Rhizopus oryzae, Penicilium chrysogenum. The enzyme production was optimized by application of the five factorial design, each at three levels. These factors are carbon sources (whey, glucose and stevia, ammonium sulfate, manganese sulfate, temperature, and pH. Pectinase concentration was measured by the Miller method. Results: The results indicate that optimum condition for enzyme production for three fungi strains was obtained at 32 °C, pH = 6, 3g / L manganese sulfate, 2.75g / L of ammonium sulfate and 10g / L of each carbon source. The best experiment in obtaining the optimum enzyme contained 1.328 mg / ml of glucose for Aspergillus niger 1.284 and 1.039 mg / ml of whey for Rhizopus oryzae and Penicilium chrysogenum. Molecular weight of enzyme was about 40 and 37 kDa which was obtained by SDS- PAGE. Discussion and conclusion: The results indicate that three strains could grow in a wide range of carbon source, pH and temperature, which could be a good candidate for industrial application.

  17. Inhibition of aflatoxin B production of Aspergillus flavus, isolated from soybean seeds by certain natural plant products.

    Science.gov (United States)

    Krishnamurthy, Y L; Shashikala, J

    2006-11-01

    The inhibitory effect of cowdung fumes, Captan, leaf powder of Withania somnifera, Hyptis suaveolens, Eucalyptus citriodora, peel powder of Citrus sinensis, Citrus medica and Punica granatum, neem cake and pongamia cake and spore suspension of Trichoderma harzianum and Aspergillus niger on aflatoxin B(1) production by toxigenic strain of Aspergillus flavus isolated from soybean seeds was investigated. Soybean seed was treated with different natural products and fungicide captan and was inoculated with toxigenic strain of A. flavus and incubated for different periods. The results showed that all the treatments were effective in controlling aflatoxin B(1) production. Captan, neem cake, spore suspension of T. harzianum, A. niger and combination of both reduced the level of aflatoxin B(1) to a great extent. Leaf powder of W. somnifera, H. suaveolens, peel powder of C. sinensis, C. medica and pongamia cake also controlled the aflatoxin B(1) production. All the natural product treatments applied were significantly effective in inhibiting aflatoxin B(1) production on soybean seeds by A. flavus. These natural plant products may successfully replace chemical fungicides and provide an alternative method to protect soybean and other agricultural commodities from aflatoxin B(1) production by A. flavus.

  18. Comparative genomics analysis of field isolates of Aspergillus flavus and A. parasiticus to explain phenotypic variation in oxidative stress tolerance and host preference

    Science.gov (United States)

    Aflatoxin contamination of peanut and other crops is a major concern for producers globally, and has been shown to be exacerbated by drought stress. Previous transcriptomic and proteomic examination of the responses of isolates of Aspergillus flavus to drought-related oxidative stress in vitro have ...

  19. Variation in fumonisin and ochratoxin production associated with differences in biosynthetic gene content in Aspergillus niger and A. welwitschiae isolates from multiple crop and geographic origins

    Science.gov (United States)

    The fungi Aspergillus niger and A. welwitschiae are morphologically indistinguishable species used for industrial fermentation and for food and beverage production. The fungi also occur widely on food crops. Concerns about their safety have arisen with the discovery that some isolates of both specie...

  20. Use of cell surface protein typing for genotyping of azole-resistant and -susceptible Aspergillus fumigatus isolates in Iran.

    Science.gov (United States)

    Falahatinejad, Mahsa; Vaezi, Afsane; Fakhim, Hamed; Abastabar, Mahdi; Shokohi, Tahereh; Zahedi, Nina; Ansari, Saham; Meis, Jacques F; Badali, Hamid

    2018-02-01

    Aspergillus fumigatus is the leading cause of mortality in severely immunocompromised individuals. Understanding pathogen dispersion and relatedness is essential for determining the epidemiology of nosocomial infections. Therefore, the aim of this study was to investigate the diversity and putative origins of clinical and environmental azole-susceptible and -resistant A. fumigatus isolates from Iran. In all, 79 isolates, including 64 azole-susceptible and 15 -resistant isolates, were genotyped using the cell surface protein (CSP) gene. Seven distinct repeat types (r01, r02, r03, r04, r05, r06 and r07) and 11 different CSP variants (t01, t02, t03, t04A, t06A, t06B, t08, t10, t18A, t18B and t22) were observed. Interestingly, t06B, t18A and t18B were exclusively present in azole-resistant isolates. The Simpson's index of diversity (D) was calculated at 0.78. Resistant isolates were genetically less diverse than azole-susceptible isolates. However, azole-resistant A. fumigatus without TR 34 /L98H were more diverse than with TR 34 /L98H. The limited CSP type diversity of the TR 34 /L98H isolates versus azole-susceptible isolates suggests that repeated independent emergence of the TR 34 /L98H mechanism is unlikely. It has been suggested that CSP types might have a common ancestor that developed locally and subsequently migrated worldwide. © 2017 Blackwell Verlag GmbH.

  1. Towards Translational ImmunoPET/MR Imaging of Invasive Pulmonary Aspergillosis: The Humanised Monoclonal Antibody JF5 Detects Aspergillus Lung Infections In Vivo

    DEFF Research Database (Denmark)

    Davies, Genna; Rolle, Anna-Maria; Maurer, Andreas

    2017-01-01

    of the fungus from invasive lung biopsy, considered the gold standard for IPA detection, is slow and often not possible in critically ill patients. In a previous study, we reported the development of a novel non-invasive procedure for IPA diagnosis based on antibody-guided positron emission tomography...... and magnetic resonance imaging (immunoPET/MRI) using a [64Cu] DOTA-labeled mouse monoclonal antibody (mAb), mJF5, specific to Aspergillus. To enable translation of the tracer to the clinical setting, we report here the development of a humanised version of the antibody (hJF5), and pre-clinical imaging of lung......Cu] NODAGA-hJF5 tracer developed here represents an ideal candidate for the diagnosis of IPA and translation to the clinical setting....

  2. Plasmodium falciparum field isolates from South America use an atypical red blood cell invasion pathway associated with invasion ligand polymorphisms

    DEFF Research Database (Denmark)

    Lopez-Perez, Mary; Villasis, Elizabeth; Machado, Ricardo L D

    2012-01-01

    members of the PfRh (PfRh1, PfRh2a, PfRh2b, PfRh4, PfRh5) families were determined. We found that most P. falciparum field isolates from Colombia and Peru invade RBCs through an atypical invasion pathway phenotypically characterized as resistant to all enzyme treatments (NrTrCr). Moreover, the invasion...... blood stage vaccine design focused on invasion pathway proteins, suggesting that regional invasion variants and global geographical variation are likely to preclude a simple one size fits all type of vaccine....

  3. Effect of gamma irradiation on growth and aflatoxin production by certain local aflatoxigenic isolates of aspergillus flavus

    International Nuclear Information System (INIS)

    Hammad, A.A.I.; Atalla, M.A.; El-Shayeb, N.M.A.; Ahmed, A.A.; Zeinat, Kamel

    1998-01-01

    The radiation D 1 0 values of two aflatoxigenic isolates of aspergillus flavus (No. 1 and No. 184) were determined. The D 1 0 -value of A. flavus isolate No.1 was found to be 0.43 and 0.50 kGy in physiological saline soulution and smoked herrings, while the D 1 0 - value of A. SFlavus isolateNo. 184 was 0.53 and -0.62 KGy in saline solution and raisins, respectively. The irradiated (0.05, 1.0, 1.5, 2.0, 2.5, 3.0 and 3.5 KGy) and non-irradiated spores of two fungi were separately inoculated into rice-corn steep liquor and incubated at 28 degree C for 12 days. The fungal mycelial dry weight, as a function of growth, decreased with increasing gamma irradiation doses. Irradiation doses used gratly reduced aflatoxins formation. Smoked herrings and raisins were inoculated with spores of A. flavus isolated No.1 and No.184, respectively, then irradiated at 1.5, 3.0 and 4.5 KGy and stored for two months at room temperature. It was found that a gamma irradiation dose of 1.5 KGy greatly reduced the amount of aflatoxins produced, while 3.0 and 4.5 KGy completely prevented aflatoxin formation

  4. First itraconazole resistant Aspergillus fumigatus clinical isolate harbouring a G54E substitution in Cyp51Ap in South America.

    Science.gov (United States)

    Leonardelli, Florencia; Theill, Laura; Nardin, María Elena; Macedo, Daiana; Dudiuk, Catiana; Mendez, Emilce; Gamarra, Soledad; Garcia-Effron, Guillermo

    A 27-year-old male rural worker was admitted with a fungal keratitis due to an injury involving plant detritus. Specimens were collected for microscopy examination and culture. The isolate was identified by morphological and molecular criteria. Susceptibility testing was performed using CLSI methods. CYP51A gene was PCR amplified and sequenced. An Aspergillus fumigatus strain resistant to itraconazole (MIC>8μg/ml) was isolated. The isolate was susceptible to amphotericin B, posaconazole, voriconazole and caspofungin. CYP51A sequencing showed two mutations leading on the G54E substitution. The patient received natamycin as treatment. This is the first report in South America of a clinical A. fumigatus strain carrying the substitution G54E at Cyp51Ap associated with itraconazole resistance. Considering the patient was azole-naive, this resistant isolate may have been acquired from the environment. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  5. Typing of Candida isolates from patients with invasive infection and concomitant colonization

    DEFF Research Database (Denmark)

    Brillowska-Dabrowska, A.; Bergmann, O.; Jensen, Irene Møller

    2010-01-01

    We investigated the relationship between colonizing and invasive isolates from patients with candidaemia. Molecular typing was performed using random amplification of polymorphic DNA (RAPD) and multilocus sequence typing (MLST). We found MLST to be sufficient for typing Candida isolates, and that......We investigated the relationship between colonizing and invasive isolates from patients with candidaemia. Molecular typing was performed using random amplification of polymorphic DNA (RAPD) and multilocus sequence typing (MLST). We found MLST to be sufficient for typing Candida isolates...

  6. Structures and antiviral activities of butyrolactone derivatives isolated from Aspergillus terreus MXH-23

    Science.gov (United States)

    Ma, Xinhua; Zhu, Tianjiao; Gu, Qianqun; Xi, Rui; Wang, Wei; Li, Dehai

    2014-12-01

    A new butyrolactone derivative, namely butyrolactone VIII ( 1), and six known butyrolactones ( 2-7) were separated from the ethyl acetate (EtOAc) extract of the fermentation broth of a fungus, Aspergillus terreus MXH-23. The chemical structures of these metabolites were identified by analyzing their nuclear magnetic resonance (NMR) and mass spectrometry (MS). Known butyrolactone derivatives contain an α, β-unsaturated γ-lactone ring with α-hydroxyl and γ-benzyl, and butyrolactone VIII ( 1) was the first butyrolactones contains α-benzyl and γ-hydroxyl on α, β-unsaturated lactone ring. All of the butyrolactone derivatives were tested for their anti-influenza (H1N1) effects. Derivatives 4 and 7 showed moderate antiviral activities while the newly-identified, derivative 1, did not.

  7. Microbial Observatory (ISS-MO): Draft Genome Sequence of two Aspergillus fumigatus Strains Isolated from the International Space Station

    Data.gov (United States)

    National Aeronautics and Space Administration — Aspergillus fumigatus is a saprophytic filamentous fungus that is ubiquitous outdoors (soil decaying vegetation) and indoors (hospitals simulated closed habitats...

  8. Metabolites with Insecticidal Activity from Aspergillus fumigatus JRJ111048 Isolated from Mangrove Plant Acrostichum specioum Endemic to Hainan Island

    Directory of Open Access Journals (Sweden)

    Zhikai Guo

    2017-12-01

    Full Text Available Fungi residing in mangroves are considered to be a bank of novel bioactive natural products. In the screening for bioactive metabolites from mangrove-derived fungi, the ethyl acetate extract of the fermentation broth of Aspergillus fumigatus JRJ111048, a fungus isolated from the leaves of the mangrove plant Acrostichum specioum endemic to Hainan island, was found to possess insecticidal activity against Spodoptera litura. Bioactivity-guided isolation lead to the discovery of seven metabolites 1–7, including one new anhydride derivative aspergide (1, one new lipid amide 11-methyl-11-hydroxyldodecanoic acid amide (2, and five known compounds; α-ethyl glucoside (3, spiculisporic acid B (4, spiculisporic acid C (5, spiculisporic acid (6, and secospiculisporic acid B (7. Their structures were established by NMR spectroscopic and MS analyses, and by comparison of previously reported data. Insecticidal activity against S. litura and antifungal activity of these compounds were investigated. As a result, the new compound 1 showed potent insecticidal activity against newly hatched larvae of S. litura, and compound 4 displayed weak antifungal activity against Candida albicans.

  9. Genetic and phenotypic diversity of naturally isolated wild strains of Aspergillus niger with hyper glucose oxidase production

    Directory of Open Access Journals (Sweden)

    MAHMOUD EL-HARIRI

    2015-12-01

    Full Text Available Glucose oxidase (GOx is the basic stone for many of biological industry worldwide. The improvement of GOx production basically depends on selection of hyper producer strain of Aspergillus niger. Selective isolation and screening for natural hyper producer strains of A. niger and sequence analysis of the GOD gene, which is responsible for production of the enzyme, are very effective approaches to investigate the naturally modified strains of A. niger with hyper productive capacity of GOx enzyme. The aims of the current study were selective isolation of naturally hyper GOx producing strains of A. niger and evaluation of their GOx activities under optimized parameters in the laboratory. Five wild Egyptian isolates of A. niger were screened for GOx and catalase activity using two types of modified basal liquid media. The GOx activity was evaluated by high throughout liquid phase system. The isolates showed a variable activity for GOx production ranged from 0 to 28.7 U.ml-1. One isolate coded Strain 7 was negative GOx producer on Vogel's broth medium in comparison to other isolates, while its GOx activity on Cazpek Dox was considered as positive (7.28 U.ml-1. It was concluded that GOx production is affected by three controllable factors – the basal media components, time of incubation, and the strain with its adaption to the media components‎. Also, the catalase activity was tested and it was produced with a different degree of variability, which may be reflected on GOx stability. GOD genes of these wild variant of A. niger were cloned and sequenced to determine intraspecies diversity of GOD between the wild variants. The comparison of isolated wild variants to other reference hyper GOx producer strains of A. niger was performed to determine if the GOD sequence analysis of these strains can be distinguished based on their GOx activity. This is the first report for isolation and detection of naturally A. niger hyper GOx-producer strains with

  10. [Guidelines for the treatment of invasive fungal disease by Aspergillus spp. and other fungi issued by the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC). 2011 Update].

    Science.gov (United States)

    Fortún, Jesús; Carratalá, Jordi; Gavaldá, Joan; Lizasoain, Manuel; Salavert, Miguel; de la Cámara, Rafael; Borges, Marcio; Cervera, Carlos; Garnacho, José; Lassaleta, Álvaro; Lumbreras, Carlos; Sanz, Miguel Ángel; Ramos, José T; Torre-Cisneros, Julián; Aguado, José M; Cuenca-Estrella, Manuel

    2011-01-01

    The guidelines on the treatment of invasive fungal disease by Aspergillus spp. and other fungi issued by the Spanish Society of Infectious Diseases and Clinical Microbiology (SEIMC) are presented. These recommendations are focused on four clinical categories: oncology-haematology patients, solid organ transplant recipients, patients admitted to intensive care units, and children. An extensive review is made of therapeutical advances and scientific evidence in these settings. These guidelines have been prepared according the SEIMC consensus rules by a working group composed of specialists in infectious diseases, clinical microbiology, critical care medicine, paediatrics and oncology-haematology. Specific recommendations on the prevention of fungal infections in these patients are included. Copyright © 2011 Elsevier España, S.L. All rights reserved.

  11. Purification and properties of two /beta/-glucosidases isolated from Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Witte, K.; Wartenberg, A.

    1989-01-01

    The cellulase complex of the fungus Aspergillus niger (strain CBS 554.65=ATCC 16 888) was fractionated by gel filtration yielding six pronounced peaks. Only proteins from the fraction corresponding to the first peak (96 kDa) showed /beta/-glucosidase activity vs. the substrate 4-nitrophenyl-/beta/-D-glucopyranoside (pNPG). These proteins have been fractionated by chromatofocusing, yielding two /beta/-glucosidases (I and II) which are shown to be homogeneous in isoelectric focusing experiments (pI=4.6 and 3.8, respectively). Kinetic experiments with pNPG, MU-glucopyranoside and cellobiose revealed that both types of /beta/-glucosidases behave like aryl-/beta/-glucosidases, /beta/-Glucosidase-I acting on pNPG exhibits a split kinetics characterized by high and low substrate-concentration kinetics which are differentiated by different values of V and of K/sub m/. In addition, /beta/-glucosidase-II is shown to be an exo-glucohydrolase as deduced from experiments with MU-cellobiopyranoside. Experimental features should be emphasized; usual soft-gel ion-exchange materials did not work in the chromatofocusing separation of the two /beta/-glucosidases, in contrast to the 10 /mu/-Si 500=DEAE exchange material (Serva) typically used in HPLC-experiments. Furthermore, protein content determinations based on different procedures yielded widely differing values. (orig.).

  12. Antifungal activity of essential oils on Aspergillus parasiticus isolated from peanuts

    Directory of Open Access Journals (Sweden)

    Yooussef Mina M.

    2016-04-01

    Full Text Available Aspergillus parasiticus is one of the most common fungi which contaminates peanuts by destroying peanut shells before they are harvested and the fungus produces aflatoxins. The aim of this study was to evaluate the antifungal activities of seventeen essential oils on the growth of the aflatoxigenic form of A. parasiticus in contaminated peanuts from commercial outlets in Georgia. The agar dilution method was used to test the antifungal activity of essential oils against this form of A. parasiticus at various concentrations: 500; 1,000; 1,500; 2,000; 2,500 ppm. Among the seventeen essential oils tested, the antifungal effect of cinnamon, lemongrass, clove and thyme resulted in complete inhibition of mycelial growth. Cinnamon oil inhibited mycelial growth at ≥ 1,000 ppm, lemongrass and clove oils at ≥ 1,500 ppm and thyme at 2,500 ppm. However, cedar wood, citronella, cumin and peppermint oils showed partial inhibition of mycelial growth. Eucalyptus oil, on the other hand, had less antifungal properties against growth of A. parasiticus, irrespective of its concentration. Our results indicate that the aflatoxigenic form of A. parasiticus is sensitive to selected essential oils, especially cinnamon. These findings clearly indicate that essential oils may find a practical application in controlling the growth of A. parasiticus in stored peanuts.

  13. ASPERGILLUS NIGER ASPERGILLUS NIGER

    African Journals Online (AJOL)

    eobe

    Additives such as low molecular weight alcohols, trace metals, phytate, lipids etc have been reported to stimulate citric acid production. Hence the objective of this study was to investigate the effect of stimulating the metabolic activity of activity of Aspergillus niger for the purpose of improved citric acid production from ...

  14. ASPERGILLUS NIGER ASPERGILLUS NIGER

    African Journals Online (AJOL)

    User

    CITRIC ACID PRODUCTION FROM DILUTE ACID HYDROLYSED CORN STARCH USING ASPERGILLUS NIGER. , N. A. Amenaghawon ..... “The effect of the sugar source on citric acid production by. Aspergillusniger”. Applied. Microbiology and Biotechnology, Vol. 19, Number 6,. 1984, pp. 393-397. [25]. Xu, D.B., Madrid ...

  15. Possible environmental origin of resistance of Aspergillus fumigatus to medical triazoles

    NARCIS (Netherlands)

    Snelders, Eveline; Huis In 't Veld, Robert A. G.; Rijs, Anthonius J. M. M.; Kema, Gert H. J.; Melchers, Willem J. G.; Verweij, Paul E.

    2009-01-01

    We reported the emergence of resistance to medical triazoles of Aspergillus fumigatus isolates from patients with invasive aspergillosis. A dominant resistance mechanism was found, and we hypothesized that azole resistance might develop through azole exposure in the environment rather than in

  16. Possible environmental origin of resistance of Aspergillus fumigatus to medical triazoles.

    NARCIS (Netherlands)

    Snelders, E.; Huis In 't Veld, R.A.; Rijs, A.J.M.M.; Kema, G.H.; Melchers, W.J.G.; Verweij, P.E.

    2009-01-01

    We reported the emergence of resistance to medical triazoles of Aspergillus fumigatus isolates from patients with invasive aspergillosis. A dominant resistance mechanism was found, and we hypothesized that azole resistance might develop through azole exposure in the environment rather than in

  17. Possible Environmental Origin of Resistance of Aspergillus fumigatus to Medical Triazoles

    NARCIS (Netherlands)

    Snelders, E.; Veld, R.; Rijs, A.; Kema, G.H.J.; Melchers, W.J.G.; Verweij, P.E.

    2009-01-01

    We reported the emergence of resistance to medical triazoles of Aspergillus fumigatus isolates from patients with invasive aspergillosis. A dominant resistance mechanism was found, and we hypothesized that azole resistance might develop through azole exposure in the environment rather than in

  18. Isolation of a Aspergillus niger lipase from a solid culture medium with aqueous two-phase systems.

    Science.gov (United States)

    Marini, Analía; Imelio, Natalia; Picó, Guillermo; Romanini, Diana; Farruggia, Beatriz

    2011-07-15

    The aim of this work is to find the best conditions to isolate lipase from a solid culture medium of Aspergillus niger NRRL3 strains using aqueous two-phase systems formed with polyethylene glycol and potassium phosphate or polyethylene glycol and sodium citrate. We studied the partitioning of a commercial lyophilizate from A. niger. Also, the lipase enzymatic activity was studied in all the phases of the systems and the results indicate that citrate anion increases lipase activity. An analysis by fluorescence spectroscopy of the interaction between lipase and the bottom and top phases of the systems shows that the protein tryptophan-environments are modified by the presence of PEG and salts. Separation of the enzyme from the rest of the proteins that make up the lyophilized was achieved with good yield and separation factor by ATPS formed by PEG 1000/Pi at pH 7, PEG 2000/Ci at pH 5.2 and PEG 4000/Ci at pH 5.2. The above mentioned systems were used in order to isolate extracellular lipase from a strain of A. niger in submerged culture and solid culture. The best system for solid culture, with high purification factor (30.50), is the PEG 4000/Ci at pH 5.2. The enzyme was produced in a solid culture medium whose production is simple and recovered in a phase poor in polymer, bottom phase. An additional advantage is that the citrate produces less pollution than the phosphate. This methodology could be used as a first step for the isolation of the extracellular lipase from A. niger. Copyright © 2011 Elsevier B.V. All rights reserved.

  19. Genetic and Toxigenic Variability within Aspergillus flavus Population Isolated from Maize in Two Diverse Environments in Kenya

    Directory of Open Access Journals (Sweden)

    Sheila Okoth

    2018-01-01

    Full Text Available Aspergillus flavus is the main producer of carcinogenic aflatoxins in agricultural commodities such as maize. This fungus occurs naturally on crops, and produces aflatoxins when environmental conditions are favorable. The aim of this study is to analyse the genetic variability among 109 A. flavus isolates previously recovered from maize sampled from a known aflatoxin-hotspot (Eastern region, Kenya and the major maize-growing area in the Rift Valley (Kenya, and to determine their toxigenic potential. DNA analyses of internal transcribed spacer (ITS regions of ribosomal DNA, partial β-tubulin gene (benA and calmodulin gene (CaM sequences were used. The strains were further analyzed for the presence of four aflatoxin-biosynthesis genes in relation to their capability to produce aflatoxins and other metabolites, targeting the regulatory gene aflR and the structural genes aflP, aflD, and aflQ. In addition, the metabolic profile of the fungal strains was unraveled using state-of-the-art LC-MS/MS instrumentation. The three gene-sequence data grouped the isolates into two major clades, A. minisclerotigenes and A. flavus. A. minisclerotigenes was most prevalent in Eastern Kenya, while A. flavus was common in both regions. A. parasiticus was represented by a single isolate collected from Rift Valley. Diversity existed within the A. flavus population, which formed several subclades. An inconsistency in identification of some isolates using the three markers was observed. The calmodulin gene sequences showed wider variation of polymorphisms. The aflatoxin production pattern was not consistent with the presence of aflatoxigenic genes, suggesting an inability of the primers to always detect the genes or presence of genetic mutations. Significant variation was observed in toxin profiles of the isolates. This is the first time that a profound metabolic profiling of A. flavus isolates was done in Kenya. Positive associations were evident for some metabolites

  20. Genetic and Toxigenic Variability within Aspergillus flavus Population Isolated from Maize in Two Diverse Environments in Kenya.

    Science.gov (United States)

    Okoth, Sheila; De Boevre, Marthe; Vidal, Arnau; Diana Di Mavungu, José; Landschoot, Sofie; Kyallo, Martina; Njuguna, Joyce; Harvey, Jagger; De Saeger, Sarah

    2018-01-01

    Aspergillus flavus is the main producer of carcinogenic aflatoxins in agricultural commodities such as maize. This fungus occurs naturally on crops, and produces aflatoxins when environmental conditions are favorable. The aim of this study is to analyse the genetic variability among 109 A. flavus isolates previously recovered from maize sampled from a known aflatoxin-hotspot (Eastern region, Kenya) and the major maize-growing area in the Rift Valley (Kenya), and to determine their toxigenic potential. DNA analyses of internal transcribed spacer (ITS) regions of ribosomal DNA, partial β-tubulin gene (benA) and calmodulin gene (CaM) sequences were used. The strains were further analyzed for the presence of four aflatoxin-biosynthesis genes in relation to their capability to produce aflatoxins and other metabolites, targeting the regulatory gene aflR and the structural genes aflP, aflD, and aflQ. In addition, the metabolic profile of the fungal strains was unraveled using state-of-the-art LC-MS/MS instrumentation. The three gene-sequence data grouped the isolates into two major clades, A. minisclerotigenes and A. flavus . A. minisclerotigenes was most prevalent in Eastern Kenya, while A. flavus was common in both regions. A. parasiticus was represented by a single isolate collected from Rift Valley. Diversity existed within the A. flavus population, which formed several subclades. An inconsistency in identification of some isolates using the three markers was observed. The calmodulin gene sequences showed wider variation of polymorphisms. The aflatoxin production pattern was not consistent with the presence of aflatoxigenic genes, suggesting an inability of the primers to always detect the genes or presence of genetic mutations. Significant variation was observed in toxin profiles of the isolates. This is the first time that a profound metabolic profiling of A. flavus isolates was done in Kenya. Positive associations were evident for some metabolites, while for

  1. Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance

    Directory of Open Access Journals (Sweden)

    Paulo S. Monteiro

    2015-03-01

    Full Text Available An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE, indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The KM for sodium phytate hydrolysis was 30.9 mM, while the kcat and kcat/KM were 1.46 ×105 s−1 and 4.7 × 106s−1.M−1, respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthylphosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg2+, Cd2+, K+ and Ca2+, and it was drastically inhibited by F−. The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in livestock feed, liberating 15.3 μmol phosphate/mL after 2.5 h of treatment.

  2. Characterisation of invasive clinical Haemophilus influenzae isolates in Queensland, Australia using whole-genome sequencing.

    Science.gov (United States)

    Staples, M; Graham, R M A; Jennison, A V

    2017-06-01

    Haemophilus influenzae is an important aetiological organism of both adult and child respiratory disease. The number of non-typeable (NTHi) invasive H. influenzae isolates referred to the Queensland (QLD) Public Health Microbiology laboratory has increased notably year-by-year. In this study we used whole-genome sequencing to molecularly characterise 100 referred invasive H. influenzae, including 74 NTHi isolates over a 15-year period, observing the carriage of capsular and putative virulence genes, including the major adhesins, antimicrobial resistance genes and population diversity. Encapsulated isolates were largely clonal, however NTHi isolates displayed high genetic variability by MLST and single nucleotide polymorphism typing with no dominant clone observed. The only mechanism for β-lactam resistance identified in the QLD isolates was β-lactamase production. No single set of virulence determinants was conclusively associated with invasive QLD NTHi isolates.

  3. Description of the Pathogenic Features of Streptococcus pyogenes Isolates from Invasive and Non-Invasive Diseases in Aichi, Japan.

    Science.gov (United States)

    Matsumoto, Masakado; Yamada, Kazuhiro; Suzuki, Masahiro; Adachi, Hirokazu; Kobayashi, Shinichi; Yamashita, Teruo; Minagawa, Hiroko; Tatsuno, Ichiro; Hasegawa, Tadao

    2016-07-22

    We identified hypervirulent Streptococcus pyogenes in 27 and 420 isolates from patients with invasive and non-invasive diseases, respectively, in Aichi Prefecture, Japan, between 2003 and 2012, in an attempt to understand why the prevalence of streptococcal toxic shock syndrome (STSS) suddenly increased in this location during 2011. Hypervirulent strains belong to the emm1 genotype, with a mutation in the covR/S genes that regulate many other genes, encoding virulence determinants and resulting in the absence of the proteinase streptococcal exotoxin B and the production of virulence factors such as the superantigen streptococcal exotoxin A, the nuclease streptococcal DNase, the cytotoxin NAD-glycohydrolase, and the hemolysin streptolysin O. We found 1 strain from invasive disease and 1 from non-invasive disease with traits similar to those of hypervirulent strains, except that the sda1 gene was absent. We also found 1 non-emm1 strain with phenotypic and genetic traits identical to those of the emm1 hypervirulent strains except that it did not belong to emm1 genotype, from non-invasive diseases cases in 2011. These findings suggested that hypervirulent and hypervirulent-like strains from invasive and non-invasive disease cases could have at least partially contributed to the sudden increase in the number of patients with STSS in Aichi during 2011.

  4. Phytosterols elevation in bamboo shoot residue through laboratorial scale solid-state fermentation using isolated Aspergillus niger CTBU.

    Science.gov (United States)

    Zheng, X X; Chen, R S; Shen, Y; Yin, Z Y

    2014-04-01

    Aspergillus niger CTBU isolated from local decayed bamboo shoot residue was employed to solid-state fermentation (SSF) of bamboo shoot residue to elevate the content of phytosterols. Strain acclimatization was carried out under the fermentation condition using bamboo shoot as substrate for fermentation performance improvement. The optimal fermentation temperature and nitrogen level were investigated using acclimatized strain, and SSF was carried out in a 500-ml Erlenmeyer flask feeding 300-mg bamboo shoot residue chips under the optimal condition (33 °C and feeding 4 % urea), and 1,186 mg (100 g)(-1) of total phytosterol was attained after 5-day fermentation, in comparison, only 523 mg (100 g)(-1) of phytosterol was assayed in fresh shoots residue. HPLC analysis of the main composition of total phytosterols displays that the types of phytosterols and composition ratio of main sterols keep steady. This laboratorial scale SSF unit could be scaled up for raw phytosterols production from discarded bamboo shoot residue and could reduce its cost.

  5. Characterization of Lignocellulolytic Activities from a Moderate Halophile Strain of Aspergillus caesiellus Isolated from a Sugarcane Bagasse Fermentation

    Science.gov (United States)

    Miranda-Miranda, Estefan; Sánchez-Reyes, Ayixón; Cuervo-Soto, Laura; Aceves-Zamudio, Denise; Atriztán-Hernández, Karina; Morales-Herrera, Catalina; Rodríguez-Hernández, Rocío; Folch-Mallol, Jorge

    2014-01-01

    A moderate halophile and thermotolerant fungal strain was isolated from a sugarcane bagasse fermentation in the presence of 2 M NaCl that was set in the laboratory. This strain was identified by polyphasic criteria as Aspergillus caesiellus. The fungus showed an optimal growth rate in media containing 1 M NaCl at 28°C and could grow in media added with up to 2 M NaCl. This strain was able to grow at 37 and 42°C, with or without NaCl. A. caesiellus H1 produced cellulases, xylanases, manganese peroxidase (MnP) and esterases. No laccase activity was detected in the conditions we tested. The cellulase activity was thermostable, halostable, and no differential expression of cellulases was observed in media with different salt concentrations. However, differential band patterns for cellulase and xylanase activities were detected in zymograms when the fungus was grown in different lignocellulosic substrates such as wheat straw, maize stover, agave fibres, sugarcane bagasse and sawdust. Optimal temperature and pH were similar to other cellulases previously described. These results support the potential of this fungus to degrade lignocellulosic materials and its possible use in biotechnological applications. PMID:25162614

  6. Enhancing taxol production in a novel endophytic fungus, Aspergillus aculeatinus Tax-6, isolated from Taxus chinensis var. mairei.

    Science.gov (United States)

    Qiao, Weichuan; Ling, Fei; Yu, Lei; Huang, Yifang; Wang, Ting

    2017-12-01

    Taxol is a curatively effective but rare anti-tumor agent extracted from Taxus (yew) barks; however, the high cost and low production of the extraction method have limited its widespread use. In this study, fungi isolated from Taxus chinensis var. mairei were tested for their ability to produce taxol. High performance liquid chromatography combined with mass spectrometry confirmed that Aspergillus aculeatinus Tax-6, one of the endophytic fungi, could produce taxol in potato dextrose agar liquid medium. NaOAc, Cu 2+ , and salicylic acid were introduced into the medium to enhance taxol production of strain Tax-6 because NaOAc is an important precursor of taxol, Cu 2+ may enhance the activity of oxidase and catalyze the formation of taxol, and salicylic acid could be an elicitor signal. Application of response surface methodology to optimize the culture led to the addition of CuSO 4 , salicylic acid and sodium acetate at 0.1 mg L -1 , 10 mg L -1 and 8 g L -1 to improve taxol yield from 334.92 to 1337.56 μg L -1 . Overall, the results of this study confirmed that fungal taxol has the potential to be broadly applied by optimizing the culture conditions. Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  7. Characterization of a Newly Isolated Marine Fungus Aspergillus dimorphicus for Optimized Production of the Anti-Tumor Agent Wentilactones

    Directory of Open Access Journals (Sweden)

    Rui Xu

    2015-11-01

    Full Text Available The potential anti-tumor agent wentilactones were produced by a newly isolated marine fungus Aspergillus dimorphicus. This fungus was derived from deep-sea sediment and identified by polyphasic approach, combining phenotypic, molecular, and extrolite profiles. However, wentilactone production was detected only under static cultures with very low yields. In order to improve wentilactone production, culture conditions were optimized using the response surface methodology. Under the optimal static fermentation conditions, the experimental values were closely consistent with the prediction model. The yields of wentilactone A and B were increased about 11-fold to 13.4 and 6.5 mg/L, respectively. The result was further verified by fermentation scale-up for wentilactone production. Moreover, some small-molecule elicitors were found to have capacity of stimulating wentilactone production. To our knowledge, this is first report of optimized production of tetranorlabdane diterpenoids by a deep-sea derived marine fungus. The present study might be valuable for efficient production of wentilactones and fundamental investigation of the anti-tumor mechanism of norditerpenoids.

  8. Production and Characterization of Lipases by Two New Isolates of Aspergillus through Solid-State and Submerged Fermentation

    Science.gov (United States)

    Colla, Luciane Maria; Ficanha, Aline M. M.; Rizzardi, Juliana; Bertolin, Telma Elita; Reinehr, Christian Oliveira; Costa, Jorge Alberto Vieira

    2015-01-01

    Due to the numerous applications of lipases in industry, there is a need to study their characteristics, because lipases obtained from different sources may present different properties. The aim of this work was to accomplish the partial characterization of lipases obtained through submerged fermentation and solid-state fermentation by two species of Aspergillus. Fungal strains were isolated from a diesel-contaminated soil and selected as good lipases producers. Lipases obtained through submerged fermentation presented optimal activities at 37°C and pH 7.2 and those obtained through solid-state fermentation at 35°C and pH 6.0. The enzymes produced by submerged fermentation were more temperature-stable than those obtained by solid-state fermentation, presenting 72% of residual activity after one hour of exposition at 90°C. Lipases obtained through submerged fermentation had 80% of stability in acidic pH and those obtained through solid-state fermentation had stability greater than 60% in alkaline pH. PMID:26180809

  9. Isolation, screening and characterization of a novel extracellular xylanase from Aspergillus niger (KP874102.1) and its application in orange peel hydrolysis.

    Science.gov (United States)

    Uday, Uma Shankar Prasad; Majumdar, Ria; Tiwari, Onkar Nath; Mishra, Umesh; Mondal, Abhijit; Bandyopadhyay, Tarun Kanti; Bhunia, Biswanath

    2017-12-01

    In the present work, a potent xylanase producing fungal strain Aspergillus niger (KP874102.1) was isolated through cultural and morphological observations from soil sample of Baramura forest, Tripura west, India. 28S rDNA technique was applied for genomic identification of this fungal strain. The isolated strain was found to be phylogenetically closely related to Aspergillus niger. Kinetic constants such as K m and V max for extracellular xylanase were determined using various substrate such as beech wood xylan, oat spelt xylan and CM cellulose through Lineweaver-Burk plot. K m , V max and K cat for beech wood xylan are found to be 2.89mg/ml, 2442U and 426178Umlmg -1 respectively. Crude enzyme did not show also CM cellulose activity. The relative efficiency of oat spelt xylan was found to be 0.819 with respect to beech wood xylan. After acid hydrolysis, enzyme was able to produce reducing sugar with 17.7, 35.5, 50.8 and 65% (w/w) from orange peel after 15, 30, 45 and 60min incubation with cellulase free xylanase and maximum reducing sugar formation rate was found to be 55.96μg/ml/min. Therefore, the Aspergillus niger (KP874102.1) is considered as a potential candidate for enzymatic hydrolysis of orange peel. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. optimization of process parameters for lovastatin production under solid-state fermentation from ground corn cobs by gamma irradiated aspergillus tamarri isolate

    International Nuclear Information System (INIS)

    Mattar, Z.A.; Khalaf, M.A.; Meleigy, S.A.

    2010-01-01

    rapid screening method is demonstrated for isolating lovastatin overproducing strains of gamma irradiated aspergillus tamarri. the screening methodology, based on the activity of lovastatin against the yeast candida albicans. among 24 gamma irradiated isolates of a. tamarri, the isolate G-8 was selected as best producer for lovastatin. solid state fermentation (SSF)was evaluated to produce lovastatin by a. tamarri G-8 isolate using ground corn cobs as substrate. monofactorial experiments were adopted to optimize the fermentation conditions. various crucial parameters such as particle size, moisture content, ph, temperature, inoculum size and incubation time were derived. corn cobs of particle size 0.4 mm having moisture level of 60 % and ph 5 gave the highest yield of lovastatin (12.4 mg/gram dry substrate) when inoculated with a. tamarri G-8 at inoculum size 10 % and 28 degree C for 8 days.

  11. Serotype and Genotype Distribution among Invasive Streptococcus pneumoniae Isolates in Colombia, 2005–2010

    Science.gov (United States)

    Parra, Eliana L.; Ramos, Viviana; Sanabria, Olga; Moreno, Jaime

    2014-01-01

    In Colombia, a laboratory-based surveillance of invasive Streptococcus pneumoniae isolates as part of SIREVA II PAHO has been conducted since 1994. This study describes the serotype distribution, antimicrobial resistance, and genetic relationships of pneumococcal isolates recovered in Colombia from 2005 to 2010. In this study, demographic data of invasive S. pneumoniae isolates were analyzed, and antimicrobial susceptibility patterns were determined. Pulse field gel electrophoresis (n = 629) and multilocus sequence typing (n = 10) were used to determine genetic relationship of isolates with minimal inhibitory concentration to penicillin ≥0.125 µg/mL. A total of 1775 isolates of S. pneumoniae were obtained. Fifteen serotypes accounted for 80.7% of isolates. Serotype 14 (23.1%) was the most frequent in the general population. Penicillin resistance was 30.7% in meningitis and 9.0% in non-meningitis. Clones Spain6BST90, Spain9VST156, Spain23FST81, and Colombia23FST338 were associated to isolates. Additionally, serotype 6A isolates were associated with ST460 and ST473, and 19A isolates with ST276, ST320, and ST1118. In conclusion, the surveillance program provided updated information of trends in serotype distribution, antimicrobial resistance and the circulation of clones in invasive pneumococcal diseases. These results could be helpful to understand the epidemiology of S. pneumoniae in Colombia, and provide a baseline to measure the impact of vaccine introduction. PMID:24416330

  12. Aspergillus-Related Lung Disease

    Directory of Open Access Journals (Sweden)

    Alia Al-Alawi

    2005-01-01

    Full Text Available Aspergillus is a ubiquitous dimorphic fungus that causes a variety of human diseases ranging in severity from trivial to life-threatening, depending on the host response. An intact host defence is important to prevent disease, but individuals with pre-existing structural lung disease, atopy, occupational exposure or impaired immunity are susceptible. Three distinctive patterns of aspergillus-related lung disease are recognized: saprophytic infestation of airways, cavities and necrotic tissue; allergic disease including extrinsic allergic alveolitis, asthma, allergic bronchopulmonary aspergillosis, bronchocentric granulomatosis and chronic eosinophilic pneumonia; and airway and tissue invasive disease -- pseudomembranous tracheobronchitis, acute bronchopneumonia, angioinvasive aspergillosis, chronic necrotizing aspergillosis and invasive pleural disease. A broad knowledge of these clinical presentations and a high index of suspicion are required to ensure timely diagnosis and treatment of the potentially lethal manifestations of aspergillus-related pulmonary disease. In the present report, the clinical, radiographic and pathological aspects of the various aspergillus-related lung diseases are briefly reviewed.

  13. Occurrence of the invasion associated marker (iam in Campylobacter jejuni isolated from cattle

    Directory of Open Access Journals (Sweden)

    Sanad Yasser M

    2011-12-01

    Full Text Available Abstract Background The invasion associated marker (iam has been detected in the majority of invasive Campylobacter jejuni retrieved from humans. Furthermore, the detection of iam in C. jejuni isolated from two important hosts, humans and chickens, suggested a role for this marker in C. jejuni's colonization of multiple hosts. However, no data exist regarding the occurrence of this marker in C. jejuni isolated from non-poultry food-animals such as cattle, an increasingly important source for human infections. Since little is known about the genetics associated with C. jejuni's capability for colonizing physiologically disparate hosts, we investigated the occurrence of the iam in C. jejuni isolated from cattle and assessed the potential of iam-containing cattle and human isolates for chicken colonization and human cell invasion. Results Simultaneous RAPD typing and iam-specific PCR analysis of 129 C. jejuni isolated from 1171 cattle fecal samples showed that 8 (6.2% of the isolates were iam-positive, while 7 (54% of human-associated isolates were iam-positive. The iam sequences were mostly heterogeneous and occurred in diverse genetic backgrounds. All iam-positive isolates were motile and possessed important genes (cadF, ciaB, cdtB associated with adhesion and virulence. Although certain iam-containing isolates invaded and survived in INT-407 cells in high numbers and successfully colonized live chickens, there was no clear association between the occurrence, allelic sequence, and expression levels of the iam and the aforementioned phenotypes. Conclusions We show that the prevalence of iam in cattle C. jejuni is relatively lower as compared to isolates occurring in humans and chickens. In addition, iam was polymorphic and certain alleles occur in cattle isolates that were capable of colonizing and invading chickens and human intestinal cells, respectively. However, the iam did not appear to contribute to the cattle-associated C. jejuni

  14. Effects of soil moisture and temperature on preharvest invasion of peanuts by the Aspergillus flavus group and subsequent aflatoxin development.

    Science.gov (United States)

    Hill, R A; Blankenship, P D; Cole, R J; Sanders, T H

    1983-02-01

    Four soil temperature and moisture treatment regimens were imposed on Florunner peanuts 94 days after planting in experimental plots in 1980. At harvest (145 days after planting), the incidence of the Aspergillus flavus group and the aflatoxin concentration were greatest in damaged kernels. Extensive colonization of sound mature kernels (SMK) by the A. flavus group occurred with the drought stress treatment (56% kernels colonized); colonization was less in the irrigated plot (7%) and the drought stress plot with cooled soil (11%) and was intermediate in the irrigated plot with heated soil (26%). Aflatoxin was virtually absent from SMK with the last three treatments, but it was found at an average concentration of 244 ppb (ng/g) in drought-stressed SMK. Colonization of SMK by the A. flavus group and aflatoxin production were greater with hot dry conditions. Neither elevated temperature alone nor drought stress alone caused aflatoxin contamination in SMK. When the ratio of SMK colonized by A. flavus compared with A. niger was greater than 19:1, there was aflatoxin contamination, but there was none if this ratio was less than 9:1. Irrigation caused a higher incidence of A. niger than drought did. This may have prevented the aflatoxin contamination of undamaged peanuts.

  15. Volatile profiles and aflatoxin production by toxigenic and non-toxigenic isolates of Aspergillus flavus grown on sterile and non-sterile cracked corn.

    Science.gov (United States)

    De Lucca, Anthony J; Boue, Stephen M; Carter-Wientjes, Carol; Bhatnagar, Deepak

    2012-01-01

    Aspergillus flavus is a saprophytic fungus which can grow on corn and produce aflatoxins which render it unsafe for consumption as food and feed. In this study, aflatoxin and non-aflatoxin producing isolates of A. flavus were grown separately on wet (20% water added), sterile or non-sterile cracked corn. Wet and dry cracked corn controls were included as needed. Secondary metabolic volatiles were identified and aflatoxin concentrations determined over a 12-day period. Volatiles unique to the toxigenic A. flavus isolates were determined by comparison with volatiles produced by the respective corn controls and the non-toxigenic A. flavus isolate. The number and identity of the volatiles produced by these A. flavus isolates varied by isolate, whether sterile or non-sterile corn was the substrate, and the sampling day. Overall, most of the volatiles were produced before day 8 after inoculation. Aflatoxin production was 10-fold lower on the sterile corn, compared to the non-sterile corn. Volatiles unique to the aflatoxin producing isolates were identified on both substrates after comparison with those produced by the non-aflatoxin producing isolate, as well as the corn control samples. Results indicate that several factors (substrate, fungal isolate, culture age) affect volatile and aflatoxin production by A. flavus.

  16. Laboratory characterization of invasive Haemophilus influenzae isolates from Nunavut, Canada, 2000-2012.

    Science.gov (United States)

    Tsang, Raymond S W; Li, Y Anita; Mullen, Angie; Baikie, Maureen; Whyte, Kathleen; Shuel, Michelle; Tyrrell, Gregory; Rotondo, Jenny A L; Desai, Shalini; Spika, John

    2016-01-01

    With invasive Haemophilus influenzae serotype b (Hib) disease controlled by vaccination with conjugate Hib vaccines, there is concern that invasive disease due to non-serotype b strains may emerge. This study characterized invasive H. influenzae (Hi) isolates from Nunavut, Canada, in the post-Hib vaccine era. Invasive H. influenzae isolates were identified by conventional methods at local hospitals; and further characterized at the provincial and federal public health laboratories, including detection of serotype antigens and genes, multi-locus sequence typing and antibiotic susceptibility. Of the 89 invasive H. influenzae cases identified from 2000 to 2012, 71 case isolates were available for study. There were 43 serotype a (Hia), 12 Hib, 2 Hic, 1 Hid, 1 Hie, 2 Hif and 10 were non-typeable (NT). All 43 Hia were biotype II, sequence type (ST)-23. Three related STs were found among the Hib isolates: ST-95 (n=9), ST-635 (n=2) and ST-44 (n=1). Both Hif belonged to ST-124 and the 2 Hic were typed as ST-9. The remaining Hid (ST-1288) and Hie (ST-18) belonged to 2 separate clones. Of the 10 NT strains, 3 were typed as ST-23 and the remaining 7 isolates each belonged to a unique ST. Eight Hib and 1 NT-Hi were found to be resistant to ampicillin due to β-lactamase production. No resistance to other antibiotics was detected. During the period of 2000-2012, Hia was the predominant serotype causing invasive disease in Nunavut. This presents a public health concern due to an emerging clone of Hia as a cause of invasive H. influenzae disease and the lack of published guidelines for the prophylaxis of contacts. The clonal nature of Hia could be the result of spread within an isolated population, and/or unique characteristics of this strain to cause invasive disease. Further study of Hia in other populations may provide important information on this emerging pathogen. No antibiotic resistance was detected among Hia isolates; a small proportion of Hib and NT-Hi isolates

  17. [Biological safety of the molluscicidal ingredient from Aspergillus fumigatus SL-30 isolated from rhizosphere of Phytolacca acinosa].

    Science.gov (United States)

    Guo, Dan-Zhao; Chen, Jun

    2012-12-30

    To study biological safety of the molluscicidal ingredient (MI, a kind of diketopiperazines) in the exocellular broth of Aspergillus fumigatus SL-30' which was isolated from the rhizosphere of Phytolacca acinosa. The MI was prepared in concentrations ranging from 0.01 to 0.60 mg/L, at the same time molluscicidal activity against Oncomelannia hupensis was tested by using immersion method, and the LC50 in the case of immersion time for 24, 48 and 72 h was calculated. Acute toxicity of the MI with concentration ranging from 0.5 to 4.0 mg/L on Brachydanio rerio, Macrobrachium nippoensis and Rana limnochris was performed by standard laboratory procedure, and mortality rate of the tested animals with treated time of 24, 48 and 72 h was recorded. Effect of the MI on Eisenia fetida was tested using natural soil procedure with the dose of 1 and 10 mg/kg, and the poisoning performance and mortality in the 7th day and 14th day were recorded. Inhibition of the MI at 10 mg/kg on respiration of soil microorganisms was determined by sealed alkaline direct absorption method, and the released amount of CO2 (mg/100 g) in the 2nd, 5th, 7th, 10th, 12th and 15th day was determined. Molluscicidal activity of the MI against Oncomelania hupensis was shown with LC50 0.101, 0.062, and 0.022 mg/L in 24, 48 and 72 h, respectively. Mortality rate of Brachydanio rerio, Macrobrachium nippoensis and Rana limnochris all increased with the increase of MI concentration, and the LC50 of each kind of tested animals was recorded as follows: 1.941, 1.755 and 1.219 mg/L for Brachydanio rerio, 3.170, 2.720 and 2.419 mg/L for Macrobrachium nippoensis, 2.109, 1.751 and 1288 mg/L for Rana limnochris in 24, 48 and 72 h, respectively. The LC50 of MI on Eisenia fetida was >10.0 mg/kg. The inhibition rate of MI was less than 50% with a concentration of 10 mg/kg in 15 days. The MI shows molluscicidal activity on Oncomelania hupensis with much lower toxicity on Brachydanio rerio, Macrobrachium nippoensis and Rana

  18. Purification, biochemical characterization and structural modelling of alkali-stable β-1,4-xylan xylanohydrolase from Aspergillus fumigatus R1 isolated from soil.

    Science.gov (United States)

    Deshmukh, Rehan Ahmed; Jagtap, Sharmili; Mandal, Madan Kumar; Mandal, Suraj Kumar

    2016-02-04

    Aspergillus fumigatus R1 produced xylanase under submerged fermentation which degrades the complex hemicelluloses contained in agricultural substrates. Xylanases have gained considerable attention because of their tremendous applications in industries. The purpose of our study was to purify xylanase and study its biochemical properties. We have predicted the secondary structure of purified xylanase and evaluated its active site residues and substrate binding sites based on the global and local structural similarity. Various microorganisms were isolated from Puducherry soil and screened by Congo-red test. The best isolate was identified to be Aspergillus fumigatus R1. The production kinetics showed the highest xylanase production (208 IU/ml) by this organism in 96 h using 1 % rice bran as the only carbon source. The purification of extracellular xylanase was carried out by fractional ammonium sulphate precipitation (30-55 %), followed by extensive dialysis and Bio-Gel P-60 Gel-filtration chromatography. The enzyme was purified 58.10 folds with a specific activity of 38196.22 IU/mg. The biochemical characterization of the pure enzyme was carried out for its optimum pH and temperature (5.0 and 50(0)C), pH and temperature stability, molecular mass (Mr) (24.5 kDa) and pI (6.29). The complete sequence of protein was obtained by mass spectrometry analysis. Apparent Km and Vmax values of the xylanase for birchwood xylan were 11.66 mg/ml and 87.6 μmol min(-1) mg(-1) respectively. Purified xylanase was analyzed by mass-spectrometry which revealed 2 unique peptides. Xylanase under current study showed significant production using agricultural residues and a broad range of pH stability in the alkaline region. Xylanase produced by Aspergillus fumigatus R1 could serve as the enzyme of choice in industries.

  19. Abundance, genetic diversity and sensitivity to demethylation inhibitor fungicides of Aspergillus fumigatus isolates from organic substrates with special emphasis on compost.

    Science.gov (United States)

    Santoro, Karin; Matić, Slavica; Gisi, Ulrich; Spadaro, Davide; Pugliese, Massimo; Gullino, Maria L

    2017-12-01

    Aspergillus fumigatus is a widespread fungus that colonizes dead organic substrates but it can also cause fatal human diseases. Aspergilloses are treated with demethylation inhibitor (DMI) fungicides; however, resistant isolates appeared recently in the medical and also environmental area. The present study aims at molecular characterizing and quantifying A. fumigatus in major environmental habitats and determining its sensitivity to medical and agricultural DMI fungicides. A. fumigatus was isolated only rarely from soil and meadow/forest organic matter but high concentrations (10 3 to 10 7  cfu/g) were detected in substrates subjected to elevated temperatures, such as compost and silage. High genetic diversity of A. fumigatus from compost was found based on SSR markers, distinguishing among fungal isolates even when coming from the same substrate sample, while subclustering was observed based on mutations in cyp51A gene. Several cyp51A amino acid substitutions were found in 15 isolates, although all isolates were fully sensitive to the tested DMI fungicides, with exception of one isolate in combination with one fungicide. This study suggests that the tested A. fumigatus isolates collected in Italy, Spain and Hungary from the fungus' major living habitats (compost) and commercial growing substrates are not potential carriers for DMI resistance in the environment. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  20. Prevalence of clonal complexes and virulence genes among commensal and invasive Staphylococcus aureus isolates in Sweden.

    Directory of Open Access Journals (Sweden)

    Gunlög Rasmussen

    Full Text Available Staphylococcus aureus encodes a remarkable number of virulence factors which may contribute to its pathogenicity and ability to cause invasive disease. The main objective of this study was to evaluate the association between S. aureus invasiveness and bacterial genotype, in terms of the presence of virulence genes and affiliation to clonal complexes. Also, the significance of different virulence genes, mainly adhesins, for the development of infective endocarditis was investigated. DNA microarray technology was used to analyze 134 S. aureus isolates, all methicillin-susceptible, derived from three groups of clinically well-characterized patients: nasal carriers (n=46, bacteremia (n=55, and bacteremia with infective endocarditis (n=33. Invasive isolates were dominant in four of the major clonal complexes: 5, 8, 15, and 25. Of the 170 virulence genes examined, those encoding accessory gene regulator group II (agr II, capsule polysaccharide serotype 5 (cap5, and adhesins such as S. aureus surface protein G (sasG and fibronectin-binding protein B (fnbB were found to be associated with invasive disease. The same was shown for the leukocidin genes lukD/lukE, as well as the genes encoding serine protease A and B (splA/splB, staphylococcal complement inhibitor (scn and the staphylococcal exotoxin-like protein (setC or selX. In addition, there was a trend of higher prevalence of certain genes or gene clusters (sasG, agr II, cap5 among isolates causing infective endocarditis compared to other invasive isolates. In most cases, the presence of virulence genes was linked to clonal complex affiliation. In conclusion, certain S. aureus clonal lineages harboring specific sets of virulence genes seem to be more successful in causing invasive disease.

  1. Standardization of a two-step real-time polymerase chain reaction based method for species-specific detection of medically important Aspergillus species

    Directory of Open Access Journals (Sweden)

    P Das

    2017-01-01

    Full Text Available Purpose: Standardization of Aspergillus polymerase chain reaction (PCR poses two technical challenges (a standardization of DNA extraction, (b optimization of PCR against various medically important Aspergillus species. Many cases of aspergillosis go undiagnosed because of relative insensitivity of conventional diagnostic methods such as microscopy, culture or antigen detection. The present study is an attempt to standardize real-time PCR assay for rapid sensitive and specific detection of Aspergillus DNA in EDTA whole blood. Materials and Methods: Three nucleic acid extraction protocols were compared and a two-step real-time PCR assay was developed and validated following the recommendations of the European Aspergillus PCR Initiative in our setup. In the first PCR step (pan-Aspergillus PCR, the target was 28S rDNA gene, whereas in the second step, species specific PCR the targets were beta-tubulin (for Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, gene and calmodulin gene (for Aspergillus niger. Results: Species specific identification of four medically important Aspergillus species, namely, A. fumigatus, A. flavus, A. niger and A. terreus were achieved by this PCR. Specificity of the PCR was tested against 34 different DNA source including bacteria, virus, yeast, other Aspergillus sp., other fungal species and for human DNA and had no false-positive reactions. The analytical sensitivity of the PCR was found to be 102 CFU/ml. Conclusion: The present protocol of two-step real-time PCR assays for genus- and species-specific identification for commonly isolated species in whole blood for diagnosis of invasive Aspergillus infections offers a rapid, sensitive and specific assay option and requires clinical validation at multiple centers.

  2. Standardization of a two-step real-time polymerase chain reaction based method for species-specific detection of medically important Aspergillus species.

    Science.gov (United States)

    Das, P; Pandey, P; Harishankar, A; Chandy, M; Bhattacharya, S; Chakrabarti, A

    2017-01-01

    Standardization of Aspergillus polymerase chain reaction (PCR) poses two technical challenges (a) standardization of DNA extraction, (b) optimization of PCR against various medically important Aspergillus species. Many cases of aspergillosis go undiagnosed because of relative insensitivity of conventional diagnostic methods such as microscopy, culture or antigen detection. The present study is an attempt to standardize real-time PCR assay for rapid sensitive and specific detection of Aspergillus DNA in EDTA whole blood. Three nucleic acid extraction protocols were compared and a two-step real-time PCR assay was developed and validated following the recommendations of the European Aspergillus PCR Initiative in our setup. In the first PCR step (pan-Aspergillus PCR), the target was 28S rDNA gene, whereas in the second step, species specific PCR the targets were beta-tubulin (for Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus), gene and calmodulin gene (for Aspergillus niger). Species specific identification of four medically important Aspergillus species, namely, A. fumigatus, A. flavus, A. niger and A. terreus were achieved by this PCR. Specificity of the PCR was tested against 34 different DNA source including bacteria, virus, yeast, other Aspergillus sp., other fungal species and for human DNA and had no false-positive reactions. The analytical sensitivity of the PCR was found to be 102 CFU/ml. The present protocol of two-step real-time PCR assays for genus- and species-specific identification for commonly isolated species in whole blood for diagnosis of invasive Aspergillus infections offers a rapid, sensitive and specific assay option and requires clinical validation at multiple centers.

  3. Diversity in Secondary Metabolites Including Mycotoxins from Strains of Aspergillus Section Nigri Isolated from Raw Cashew Nuts from Benin, West Africa.

    Science.gov (United States)

    Lamboni, Yendouban; Nielsen, Kristian F; Linnemann, Anita R; Gezgin, Yüksel; Hell, Kerstin; Nout, Martinus J R; Smid, Eddy J; Tamo, Manuele; van Boekel, Martinus A J S; Hoof, Jakob Blæsbjerg; Frisvad, Jens Christian

    2016-01-01

    In a previous study, raw cashew kernels were assayed for the fungal contamination focusing on strains belonging to the genus Aspergillus and on aflatoxins producers. These samples showed high contamination with Aspergillus section Nigri species and absence of aflatoxins. To investigate the diversity of secondary metabolites, including mycotoxins, the species of A. section Nigri may produce and thus threaten to contaminate the raw cashew kernels, 150 strains were isolated from cashew samples and assayed for their production of secondary metabolites using liquid chromatography high resolution mass spectrometry (LC-HRMS). Seven species of black Aspergilli were isolated based on morphological and chemical identification: A. tubingensis (44%), A. niger (32%), A. brasiliensis (10%), A. carbonarius (8.7%), A. luchuensis (2.7%), A. aculeatus (2%) and A. aculeatinus (0.7%). From these, 45 metabolites and their isomers were identified. Aurasperone and pyranonigrin A, produced by all species excluding A. aculeatus and A. aculeatinus, were most prevalent and were encountered in 146 (97.3%) and 145 (95.7%) isolates, respectively. Three mycotoxins groups were detected: fumonisins (B2 and B4) (2.7%) ochratoxin A (13.3%), and secalonic acids (2%), indicating that these mycotoxins could occur in raw cashew nuts. Thirty strains of black Aspergilli were randomly sampled for verification of species identity based on sequences of β-tubulin and calmodulin genes. Among them, 27 isolates were positive to the primers used and 11 were identified as A. niger, 7 as A. tubingensis, 6 as A. carbonarius, 2 as A. luchuensis and 1 as A. welwitschiae confirming the species names as based on morphology and chemical features. These strains clustered in 5 clades in A. section Nigri. Chemical profile clustering also showed also 5 groups confirming the species specific metabolites production.

  4. Co-inoculation of aflatoxigenic and non-aflatoxigenic strains of Aspergillus flavus to study fungal invasion, colonization, and competition in maize kernels

    Directory of Open Access Journals (Sweden)

    Zuzana eHruska

    2014-03-01

    Full Text Available A currently utilized pre-harvest biocontrol method involves field inoculations with non-aflatoxigenic Aspergillus flavus strains, a tactic shown to strategically suppress native aflatoxin-producing strains and effectively decrease aflatoxin contamination in corn. The present in situ study focuses on tracking the invasion and colonization of an aflatoxigenic A. flavus strain (AF70, labeled with green fluorescent protein (GFP, in the presence of a non-aflatoxigenic A. flavus biocontrol strain (AF36 to better understand the competitive interaction between these two strains in seed tissue of corn (Zea mays. Corn kernels that had been co-inoculated with GFP-labeled AF70 and wild-type AF36 were cross-sectioned and observed under UV and blue light to determine the outcome of competition between these strains. After imaging, all kernels were analyzed for aflatoxin levels. There appeared to be a population difference between the co-inoculated AF70-GFP+AF36 and the individual AF70-GFP tests, both visually and with pixel count analysis. The GFP allowed us to observe that AF70-GFP inside the kernels was suppressed up to 82% when co-inoculated with AF36 indicating that AF36 inhibited progression of AF70-GFP. This was in agreement with images taken of whole kernels where AF36 exhibited a more robust external growth compared to AF70-GFP. The suppressed growth of AF70-GFP was reflected in a corresponding (up to 73% suppression in aflatoxin levels. Our results indicate that the decrease in aflatoxin production correlated with population depression of the aflatoxigenic fungus by the biocontrol strain supporting the theory of competitive exclusion through robust propagation and fast colonization by the non-aflatoxigenic fungus.

  5. A Comparison of Aspergillus and Mucorales PCR Testing of Different Bronchoalveolar Lavage Fluid Fractions from Patients with Suspected Invasive Pulmonary Fungal Disease.

    Science.gov (United States)

    Springer, Jan; White, P Lewis; Kessel, Johanna; Wieters, Imke; Teschner, Daniel; Korczynski, Daniel; Liebregts, Tobias; Cornely, Oliver A; Schwartz, Stefan; Elgeti, Thomas; Meintker, Lisa; Krause, Stefan W; Posso, Raquel B; Heinz, Werner J; Fuhrmann, Sandra; Vehreschild, Jörg Janne; Einsele, Hermann; Rickerts, Volker; Loeffler, Juergen

    2018-02-01

    In patients with hematological malignancies, bronchoalveolar lavage fluid (BALF) specimens are commonly used for the diagnosis of mold infections. However, it is not clear whether the cell pellet (P) or the supernatant fraction (S) of the BALF specimen is optimal for molecular diagnostic testing. Thus, 99 BALF specimens were collected from 96 hematology patients with or without allogeneic hematopoietic stem cell transplant. The cell pellets and supernatants were processed alone and in combination (S/P) for testing by two fungus-specific real-time PCR assays compliant with international recommendations. The results achieved with S/P were revealed to be superior in comparison to those achieved with S and P alone, with the use of each single fraction showing a reduced sensitivity for the detection of Aspergillus DNA (82% and 43% for S and P, respectively). In 57% of the samples, testing of the combination of S and P generated a lower quantification cycle value than testing of S or P alone. Molds would have been missed in 5 and 16 out of 28 samples if only S or P, respectively, was analyzed. No sample was positive by testing of S or P only. Similar results were obtained for the detection of Mucorales DNA in BALF specimens (reduced sensitivity of 67% and 50% for S and P, respectively). Study patients were categorized according to the current European Organization for the Research and Treatment of Cancer/Mycoses Study Group classification for invasive fungal disease (IFD), revealing that 35 patients had proven/probable IFD (36%), 47 patients had possible IFD (49%), and 14 patients had undetermined IFD (15%). Copyright © 2018 American Society for Microbiology.

  6. Aflatoxins produced by Aspergillus nomius ASR3, a pathogen isolated from the leaf-cutter ant Atta sexdens rubropilosa

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    Eduardo Afonso da Silva-Junior

    Full Text Available ABSTRACT Aspergillus spp. cause economic impacts due to aflatoxins production. Although the toxicity of aflatoxins is already known, little information about their ecological roles is available. Here we investigated the compounds produced by Aspergillus nomius ASR3 directly from a dead leaf-cutter queen ant Atta sexdens rubropilosa and the fungal axenic culture. Chemical analyses were carried out by high-resolution mass spectrometry and tandem mass spectrometry techniques. Aflatoxins B1 and G1 were detected in both the axenic culture and in the dead leaf-cutter queen ant. The presence of these mycotoxins in the dead leaf-cutter queen ant suggests that these compounds can be related to the insect pathogenicity of A. nomius against A. sexdens rubropilosa.

  7. Aspergillus niger contains the cryptic phylogenetic species A. awamori

    DEFF Research Database (Denmark)

    Perrone, Giancarlo; Stea, Gaetano; Epifani, Filomena

    2011-01-01

    Aspergillus section Nigri is an important group of species for food and medical mycology, and biotechnology. The Aspergillus niger ‘aggregate’ represents its most complicated taxonomic subgroup containing eight morphologically indistinguishable taxa: A. niger, Aspergillus tubingensis, Aspergillus...... acidus, Aspergillus brasiliensis, Aspergillus costaricaensis, Aspergillus lacticoffeatus, Aspergillus piperis, and Aspergillus vadensis. Aspergillus awamori, first described by Nakazawa, has been compared taxonomically with other black aspergilli and recently it has been treated as a synonym of A. niger....... Phylogenetic analyses of sequences generated from portions of three genes coding for the proteins β-tubulin (benA), calmodulin (CaM), and the translation elongation factor-1 alpha (TEF-1α) of a population of A. niger strains isolated from grapes in Europe revealed the presence of a cryptic phylogenetic species...

  8. The production and activity test of cellulases using bagasse substrate on Aspergillus niger isolated from Clove field, Kare, Madiun

    Science.gov (United States)

    Ardhi, Muh. Waskito; Sulistyarsi, Ani; Pujiati

    2017-06-01

    Aspergillus sp is a microorganism which has a high ability to produce cellulase enzymes. In producing Cellulase enzymes requires appropriate concentration and incubation time to obtain optimum enzyme activity. This study aimed to determine the effect of inoculum concentration and incubation time towards production and activity of cellulases from Aspergillus sp substrate bagasse. This research used experiments method; completely randomized design with 2 factorial repeated 2 times. The treatment study include differences inoculum (K) 5% (K1), 15% (K2) 25%, (K3) and incubation time (F) that is 3 days (F1), 6 days (F2), 9 days (F3), 12 days (F4). The data taken from the treatment are glucose reduction and protein levels of crude cellulase enzyme activity that use Nelson Somogyi and Biuret methods. Analysis of variance ANOVA data used two paths with significance level of 5% then continued with LSD test. The results showed that: Fhit>Ftab. Thus, there is effect of inoculum concentrations and incubation time toward activity of crude cellulases of Aspergillus sp. The highest glucose reduction of treatment is K3F4 (concentration of inoculum is 25% with 12 days incubation time) amount 12.834 g / ml and the highest protein content is K3F4 (concentration of inoculum is 25% with with 12 days incubation time) amount 0.740 g / ml.

  9. Adhesive and invasive capacities of Edwarsiella tarda isolated from South American sea lion

    Directory of Open Access Journals (Sweden)

    Araceli Fernández

    2014-09-01

    Full Text Available Edwarsiella tarda is a zoonotic bacterium that can be isolated from humans, animals and the environment. Although E. tarda is primarily considered a fish pathogen, it is the only species of its genus considered to be pathogenic for humans as well. A survey of zoonotic intestinal bacteria in fresh feces from South American sea lions (SASL Otaria flavescens, reported E. tarda as the most frequently isolated species. In this study, we used HEp-2 cells to establish in vitro the adherence and invasive ability of 17 E. tarda strains isolated from SASL fecal material. All the strains were able to adhere and invade HEp-2 cells with adhesion and invasion percentages ranging from 56 to 100% and 21 to 74%, respectively. Despite the expression of these pathogenic factors, further investigation is needed to determine whether this bacterium could play a role as primary pathogen for this and other species of pinnipeds.

  10. MICROBIOLOGICAL CHARACTERISATION OF Haemophilus influenzae STRAINS ISOLATED FROM PATIENTS WITH INVASIVE AND RESPIRATORY DISEASES

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    Tomislav Kostyanev

    2010-01-01

    Full Text Available A total of 175 H. influenzae strains were collected between 1994 and 2009 from all aged patient groups. The strains were isolated from patients with invasive and community-acquired respiratory tract infections. All strains were identified according to standard microbiological methods. Serotyping was done by a coagglutination test and by molecular PCR capsular genotyping. Beta-lactamase production was determined by the chromogenic cephalosporin test with nitrocephin as substrate. Most of the isolated H. influenzae strains were from children under 5 years of age (57.7%. Overall, 61 strains belonged to serotype b (34.9% by the means of PCR capsular typing, 1 strain was type f, and 113 isolates (64.6% were non-typeable (non-encapsulated H. influenzae. Among the infants and children with meningitis or other invasive infections, aged 2 month to 5 years, all strains, except one, were serotype b. In respiratory tract infections (pneumonia, otitis media, sinusitis and people with chronic pulmonary diseases - exacerbations of COPD, bronchiectasis, cystic fibrosis the most common - 96.5% were non-typeable strains in both groups children and adults. Overall, the prevalence of beta-lactamase production was 19.4%. But, it was much higher for invasive strains from CSF isolates - 37.7%, 25% in blood samples, and 37.5% in otitis media causative strains. Beta-lactamase production was less frequent in respiratory tract isolates - in sputum 13.3% and in URT samples - 2.3%. The rate of beta-lactamase production in CSF isolates has not changed for the last 10 years.PCR capsular genotyping method has to be performed for all non-b-type strains. The implementation of Hib vaccine in our country will be accompanied by a reduction in invasive diseases caused by H. influenzae type b in children, but it is not useful in preventing infections caused by non-typeable H. influenzae strains.

  11. Invasive Ductal Carcinoma Arising in Phyllodes Tumor With Isolated Tumor Cells in Sentinel Lymph Node

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    Ying-Ju Kuo

    2010-11-01

    Full Text Available Phyllodes tumor (PT consists of stroma of variable grading and benign ductal epithelium. Although exceptional, carcinomas that arise from the epithelium in PTs do exist, and seem to behave less aggressively than the usually encountered breast carcinoma. To the best of our knowledge, among the invasive carcinomas that have arisen in PTs, only 1 has been proved to have metastatic carcinoma in the lymph nodes. Here, we describe the youngest woman to have invasive ductal carcinoma that arose in a borderline PT, with isolated carcinoma cells in the sentinel lymph node. Whether such a combined lesion carries a more indolent course is also discussed.

  12. Isolation and Characterization of Two Ammonium Permease Genes, meaA and mepA, from Aspergillus nidulans

    OpenAIRE

    Monahan, Brendon J.; Fraser, James A.; Hynes, Michael J.; Davis, Meryl A.

    2002-01-01

    Ammonium and the analogue methylammonium are taken into the cell by active transport systems which constitute a family of transmembrane proteins that have been identified in fungi, bacteria, plants, and animals. Two genes from Aspergillus nidulans, mepA and meaA, which encode ammonium transporters with different affinities have been characterized. The MepA transporter exhibits the highest affinity for methylammonium (Km0, 44.3 μM); in comparison, the Km for MeaA is 3.04 mM. By use of targeted...

  13. Effects of temperature, water activity and incubation time on fungal growth and aflatoxin B1 production by toxinogenic Aspergillus flavus isolates on sorghum seeds.

    Science.gov (United States)

    Lahouar, Amani; Marin, Sonia; Crespo-Sempere, Ana; Saïd, Salem; Sanchis, Vicente

    2016-01-01

    Sorghum, which is consumed in Tunisia as human food, suffers from severe colonization by several toxigenic fungi and contamination by mycotoxins. The Tunisian climate is characterized by high temperature and humidity that stimulates mold proliferation and mycotoxin accumulation in foodstuffs. This study investigated the effects of temperature (15, 25 and 37°C), water activity (aw, between 0.85 and 0.99) and incubation time (7, 14, 21 and 28 d) on fungal growth and aflatoxin B1 (AFB1) production by three Aspergillus flavus isolates (8, 10 and 14) inoculated on sorghum grains. The Baranyi model was applied to identify the limits of growth and mycotoxin production. Maximum diameter growth rates were observed at 0.99 a(w) at 37°C for two of the isolates. The minimum aw needed for mycelial growth was 0.91 at 25 and 37°C. At 15°C, only isolate 8 grew at 0.99 a(w). Aflatoxin B1 accumulation could be avoided by storing sorghum at low water activity levels (≤0.91 a(w)). Aflatoxin production was not observed at 15°C. This is the first work on the effects of water activity and temperature on A. flavus growth and AFB1 production by A. flavus isolates on sorghum grains. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  14. (+)-Geodin from Aspergillus terreus

    DEFF Research Database (Denmark)

    Rønnest, Mads Holger; Nielsen, Morten Thrane; Leber, Blanka

    2011-01-01

    The fungal metabolite (+)-geodin [systematic name: (2R)-methyl 5,7-dichloro-4-hydroxy-6'-methoxy-6-methyl-3,4'-dioxospiro[benzofuran-2,1'-cyclohexa-2',5'-diene]-2'-carboxylate], C(17)H(12)Cl(2)O(7), was isolated from Aspergillus terreus. The crystal structure contains two independent molecules in...... molecules. The structurally related (+)-griseofulvin has an S configuration at the spiro centre, a difference of potential biological and biosynthetic relevance....

  15. Minimally Invasive Plate Osteosynthesis In The Treatment Of Isolated Ulnar Bone Fractures

    Directory of Open Access Journals (Sweden)

    Suman K Shrestha

    2012-11-01

    Full Text Available BACKGROUND: Minimally invasive internal fixation is also called “biological internal fixation”. This concept is used widely in the treatment of various fractures but to date, there have been no report about such application in the repair of isolated ulnar bone fractures. METHODS: Eleven patients with fracture of isolated ulnar bone (four AO type 22A11 and seven AO type 22A12, mean age 43.16 (range, 24- 59y, were treated using closed reduction and locking compression plate with minimally invasive plate osteosynthesis (MIPO with the aim of minimising soft tissue damage. RESULTS: Fractures healed at an average of 7.8 weeks with good to excellent clinical outcomes. There were no complications such as nonunion, implant failure or neurovascular injuries. CONCLUSIONS: MIPO seems to be advantageous for soft tissue and bone biology. Good union was seen and fracture complications were also prevented by early mobilisation.

  16. Production of 3,4-dihydroxy L-phenylalanine by a newly isolated Aspergillus niger and parameter significance analysis by Plackett-Burman design.

    Science.gov (United States)

    Ali, S; Haq, I

    2010-12-10

    The amino acid derivative 3,4-dihydroxy L-phenylalanine (L-dopa) is gaining interest as a drug of choice for Parkinson's disease. Aspergillus oryzae is commonly used for L-dopa production; however, a slower growth rate and relatively lower tyrosinase activity of mycelia have led to an increasing interest in exploiting alternative fungal cultures. In the present investigation, we report on the microbiological transformation of L-tyrosine to L-dopa accomplished by a newly isolated filamentous fungus Aspergillus niger. The culture A. niger (isolate GCBT-8) was propagated in 500 ml Erlenmeyer flasks and the pre-grown mycelia (48 h old) were used in the reaction mixture as a source of enzyme tyrosinase. Grinded mycelia gave 1.26 fold higher L-dopa production compared to the intact at 6% glucose (pH 5.5). The rate of L-tyrosine consumption was improved from 0.198 to 0.281 mg/ml. Among the various nitrogen sources, 1.5% peptone, 1% yeast extract and 0.2% ammonium chloride were optimized. The maximal L-dopa was produced (0.365 mg/ml) at 0.3% potassium dihydrogen phosphate with L-tyrosine consumption of 0.403 mg/ml. Over ~73% yield was achieved (degree of freedom 3) when the process parameters were identified using 2k-Plackett-Burman experimental design. The results are highly significant (p ≤ 0.05) and mark the commercial utility (LSD 0.016) of the mould culture which is perhaps the first ever report on L-dopa production from A. niger.

  17. Impact of carbapenem heteroresistance among clinical isolates of invasive Escherichia coli in Chongqing, southwestern China.

    Science.gov (United States)

    Sun, J D; Huang, S F; Yang, S S; Pu, S L; Zhang, C M; Zhang, L P

    2015-05-01

    Although heteroresistance is common in a wide range of microorganisms, carbapenem heteroresistance among invasive Escherichia coli infections has not been reported. The objective of this study was to evaluate the clinical significance of carbapenem heteroresistance and to identify risk factors for its acquisition. A case-control study was conducted at a 3200-bed teaching hospital in Chongqing, southwestern China. Successive and non-duplicate nosocomial E. coli isolates (n = 332) were obtained from July 2011 to June 2013. Bloodstream isolates made up 50.6% of the strains collected. The rates of heteroresistance were 25.0% to imipenem, 17.2% to ertapenem, and 3.9% to meropenem. The population analysis profile revealed the presence of subpopulations with higher carbapenem resistance, showing MICs ranging from 2.0-128.0mg/L. Male gender, invasive intervention, antibiotic use and bacterial extended-spectrum β-lactamase (ESBL) production contributed to invasive infections by carbapenem-heteroresistant E. coli (CHEC). The production of ESBL was identified as the common independent risk factor for heteroresistance to both ertapenem and imipenem. Pulsed-field gel electrophoresis revealed clonal diversity among the CHEC isolates. Most importantly, characterization of two successive E. coli strains isolated from the same patient indicated that carbapenem resistance evolved from heteroresistance. In conclusion, the high prevalence of heteroresistance to carbapenem among invasive E. coli merits great attention. Routine detection of ESBLs and the prudent use of imipenem and ertapenem are advocated. The early targeted intervention should be formulated to reduce CHEC infection and carbapenem resistance of E. coli. Copyright © 2014 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  18. A prospective survey of Aspergillus spp. in respiratory tract samples: prevalence, clinical impact and antifungal susceptibility

    DEFF Research Database (Denmark)

    Mortensen, K L; Johansen, H K; Fuursted, Kurt

    2011-01-01

    for routine microbiologic investigation were examined for Aspergillus following routine procedures and with extended incubation (5 days). Identification was done by morphologic criteria and susceptibility testing using EUCAST method for azoles and amphotericin B E-test. Invasive aspergillosis (IA......) was evaluated using modified EORTC/MSG criteria. A total of 11,368 airway samples were received. Growth of Aspergillus spp. was found in 129 and 151 patients using routine and extended incubation, respectively. Three patients had proven IA (2%), 11 probable (7%), four had allergic bronchopulmonary aspergillosis...... μg/ml (3/112 A. fumigatus, 1/2 A. terreus). In conclusion, Aspergillus appears to be an important pathogen in Denmark. Elevated itraconazole MICs were detected in 4% of the isolates including a multi-azole resistant isolate....

  19. Density and molecular epidemiology of Aspergillus in air and relationship to outbreaks of Aspergillus infection

    NARCIS (Netherlands)

    A.C.A.P. Leenders (Alexander); A.F. van Belkum (Alex); M.D. Behrendt (Myra); A. Luijendijk (Ad); H.A. Verbrugh (Henri)

    1999-01-01

    textabstractAfter five patients were diagnosed with nosocomial invasive aspergillosis caused by Aspergillus fumigatus and A. flavus, a 14-month surveillance program for pathogenic and nonpathogenic fungal conidia in the air within and outside the University Hospital in

  20. Biofilm formation in invasive Staphylococcus aureus isolates is associated with the clonal lineage.

    Science.gov (United States)

    Naicker, Preneshni R; Karayem, Karayem; Hoek, Kim G P; Harvey, Justin; Wasserman, Elizabeth

    2016-01-01

    The contribution of the genetic background of Staphylococcus aureus to biofilm formation is poorly understood. We investigated the association between the genetic background and the biofilm forming ability of clinical invasive S. aureus isolates. Secondary objectives included investigating any correlation with biofilm formation and methicillin resistance or the source of bacteraemia. The study was conducted at a 1300-bed tertiary hospital in Cape Town, South Africa. S. aureus isolates obtained from blood cultures between January 2010 and January 2012 were included. Genotypic characterization was performed by PFGE, spa typing, SCCmec typing and MLST. Thirty genotypically unique strains were assessed for phenotypic biofilm formation with the microtitre plate assay. All isolates were tested in triplicate and an average optical density, measured at a wavelength of 490 nm, was determined. The biofilm forming ability of isolates with A490 ≤ 0.17 were considered non-adherent, A490 > 0.17 'weak positive' and A490 > 0.34 'strong positive'. Fifty seven percent of isolates formed biofilms. Weak biofilm formation occurred in 40% (n = 12) and strong biofilm formation in 17% (n = 5) of isolates. All 5 isolates capable of strong biofilm formation belong to one spa clonal complex (spa-CC 064). Strains from spa-CC 064 were capable of higher biofilm formation than other spa clonal complexes (p = 0.00002). These 5 strains belonged to MLST CC5 and CC8. Biofilm formation correlates with the spa clonal lineage in our population of invasive S. aureus strains. Biofilm formation did not correlate with methicillin resistance and was not related to the source of bacteraemia. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Detection of invasive protein profile of Streptococcus pyogenes M1 isolates from pharyngitis patients.

    Science.gov (United States)

    Hasegawa, Tadao; Okamoto, Akira; Kamimura, Takuya; Tatsuno, Ichiro; Hashikawa, Shin-Nosuke; Yabutani, Mitsutaka; Matsumoto, Masakado; Yamada, Keiko; Isaka, Masanori; Minami, Masaaki; Ohta, Michio

    2010-03-01

    Streptococcal toxic shock syndrome (STSS) is a re-emerging infectious disease in Japan and many other developed countries. Epidemiological studies have revealed that the M1 serotype of Streptococcus pyogenes is the most dominant causative isolate of STSS. Recent characterization of M1 isolates revealed that the mutation of covS, one of the two-component regulatory systems, plays an important role in STSS by altering protein expression. We analyzed the M1 S. pyogenes clinical isolates before or after 1990 in Japan, using two-dimensional gel electrophoresis (2-DE) and pulsed-field gel electrophoresis (PFGE). PFGE profiles were different between the isolates before and after 1990. Markedly different profiles among isolates after 1990 from STSS and pharyngitis patients were detected. Sequence analysis of two-component regulatory systems showed that covS mutations were detected not only in STSS but also in three pharyngitis isolates, in which proteins from the culture supernatant displayed the invasive type. The mutated CovS detected in the pharyngitis isolates had impaired function on the production of streptococcal pyrogenic exotoxin B (SpeB) analyzed by 2-DE. These results suggest that several covS mutations that lead to the malfunction of the CovS protein occurred even in pharyngeal infection.

  2. Infectious keratitis caused by Aspergillus tubingensis

    NARCIS (Netherlands)

    Kredics, L.; Varga, J.; Kocsube, S.; Rajaraman, R.; Raghavan, A.; Doczi, I.; Bhaskar, M.; Nemeth, T.M.; Antal, Z.; Venkatapathy, N.; Vagvolgyi, C.; Samson, R.A.; Chockaiya, M.; Palanisamy, M.

    2009-01-01

    PURPOSE: To report 2 cases of keratomycosis caused by Aspergillus tubingensis. METHODS: The therapeutic courses were recorded for 2 male patients, 52 and 78 years old, with fungal keratitis caused by black Aspergillus strains. Morphological examination of the isolates was carried out on malt extract

  3. Molecular characterization of Aspergillus infections in an Iranian educational hospital using RAPD-PCR method

    Directory of Open Access Journals (Sweden)

    Kambiz Diba

    2014-09-01

    Conclusion: The hospital sources for the Aspergillus clinical isolates included air condition and walls. RAPD-PCR analysis can play a trivial role to find the hospital sources of Aspergillus clinical isolates.

  4. Extracellular cellulolytic enzyme system of Aspergillus japonicus. Pt. 3. Isolation, purification and characterization of multiple forms of endoglucanase

    Energy Technology Data Exchange (ETDEWEB)

    Kundu, R.K.; Dube, Syamalima; Dube, D.K.

    1988-02-01

    The crude culture filtrate of Aspergillus japonicus exhibits marked cellulose and xylan degrading activities. Affinity chromatography of the crude enzyme preparation on concanavalin A (Con A)-Sepharose 4B resolves it into three fractions. The eluted fraction A contains CMCase (CM Case I) and xylanase activities; fraction B shows only CMCase activity (CMCase II); and fraction C exhibits CMCase (CMCase III) and xylanase activities. On further purification by gel filtration, only fraction B imparts a homogeneous preparation that gives a single band on polyacrylamide gel electrophoresis at pH 8.3. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of this homogeneous form shows the molecular weight about 57 000 daltons. The first two forms have the least activity toward tamarind kernel polysaccharide (TKP) but the last form, endoglucanase III, is highly reactive to it.

  5. Bioremediation of Synthetic and Industrial Effluents by Aspergillus niger Isolated from Contaminated Soil Following a Sequential Strategy.

    Science.gov (United States)

    Gulzar, Tahsin; Huma, Tayyaba; Jalal, Fatima; Iqbal, Sarosh; Abrar, Shazia; Kiran, Shumaila; Nosheen, Sofia; Hussain, Waqar; Rafique, Muhammad Asim

    2017-12-16

    The present study aimed to assess and compare the ability to remediate synthetic textile and industrial wastewaters by Fenton treatment, a biological system and sequential treatments using Aspergillus niger ( A. niger ). All studied treatments were found to be effective in decolorization of the effluents under study. Fenton treatment followed by A. niger showed excellent potential for the maximum decolorization of the synthetic and industrial effluents under study. The effectiveness of sequential treatment was evaluated by water quality parameters such as total organic carbon (TOC), Biological Oxygen Demand (BOD5) and Chemical Oxygen Demand (COD) before and after each treatment. The results indicated that A. niger is an effective candidate for detoxification of textile wastewaters.

  6. Serotypes and antibiotic susceptibility of Streptococcus pneumoniae isolates causative of invasive diseases in Mexican children.

    Science.gov (United States)

    Arredondo-García, José Luis; Calderón, Ernesto; Echániz-Aviles, Gabriela; Soto-Noguerón, Araceli; Arzate, Patricia; Amabile-Cuevas, Carlos F

    2011-03-02

    Streptococcus pneumoniae is a worldwide leading cause of morbidity and mortality, while susceptibility towards penicillin and macrolides can be less than 50% in many regions. A total of 150 isolates of S. pneumoniae causative of invasive diseases in children were characterized, of which 24.6% had a fatal outcome. The most prevalent serotypes were 19F, 6B, 23F and 14. Resistance to penicillin, erythromycin (mostly of macrolide-lincosamide-streptogramin resistance phenotype) or trimethoprim-sulfamethoxazole was found in more than 40% of the isolates, but no resistance phenotype appeared linked to lethality. Serotype 3 isolates, which were seldom resistant, had a twofold lethality rate compared to the total sample. Serotyping could provide a better outcome-predicting tool than susceptibility testing. The seven-valent vaccine does not include the most prevalent serotypes found in Mexico.

  7. Microsatellite analysis and susceptibility to FCZ of Candida glabrata invasive isolates in Sfax Hospital, Tunisia.

    Science.gov (United States)

    Abbes, S; Sellami, H; Sellami, A; Makni, F; Mahfoudh, N; Makni, H; Khaled, S; Ayadi, A

    2011-01-01

    We have noted that, during the last few years, there has been a redistribution of the most common Candida species with an increase in non-C. albicans Candida species, particularly Candida glabrata. In many countries, the high frequency of Candida glabrata shows the highest resistance rates. The main objective of this investigation was to analyze the genotypic variability of invasive C. glabrata isolates recovered over a period of six years and assess their in vitro susceptibility to fluconazole to determine the possible existence of relationships between genotype and susceptibility. We collected 50 invasive C. glabrata isolates (21.4%) from January 2001 to December 2007. The in vitro susceptibility profiles as determined by the E-test method showed that 8.3% of the isolates were resistant to fluconazole. The typing with three microsatellite markers RPM2, MTI and ERG3 demonstrated 12 multilocus genotypes distributed irregularly with a predominance of G1 (38%). A cluster (G9) was found among isolates collected in the same ward, at the same time period, suggesting cross transmission. Eleven of 13 patients who had previously been colonized by C. glabrata, were infected by their colonizing strains. However, we noted after prolonged treatment with fluconazole that there was an increase of the MIC for an isolate from one patient and in another patient, the selection of a more resistant variant. In our study, we didn't find an association between genotype and susceptibility to fluconazole. In conclusion, the predominance of some genotypes could be explained by nosocomial transmission or a selective ecological advantage rather than an emergence of a resistant isolate.

  8. The anti-invasive effect of lucidenic acids isolated from a new Ganoderma lucidum strain.

    Science.gov (United States)

    Weng, Chia-Jui; Chau, Chi-Fai; Chen, Kuang-Dee; Chen, Deng-Hai; Yen, Gow-Chin

    2007-12-01

    Ganoderma lucidum is a well-known mushroom with various pharmacological effects that has been used for health and longevity purposes. The objective of this study was to investigate the anti-invasive effect of lucidenic acids isolated from a new G. lucidum strain (YK-02) against human hepatoma carcinoma (HepG(2)) cells. Triterpenoid components in the ethanol extract of G. lucidum (YK-02) were separated by means of a semi-preparative RP HPLC. Four major peaks were separated and crystallized from triterpenoids fraction, and were identified as lucidenic acids A, B, C, and N according to their spectroscopic values of (1)H NMR and MS. Treatment of the lucidenic acids (50 microM) in the presence of 200 nM phorbol 12-myristate 13-acetate (PMA) after 24 h of incubation all resulted in significant inhibitory effects on PMA-induced MMP-9 activity and invasion of HepG(2 )cells. The results indicate that the lucidenic acids isolated from G. lucidum (YK-02) are anti-invasive bioactive components on hepatoma cells.

  9. The successful use of amphotericin B followed by oral posaconazole in a rare case of invasive fungal sinusitis caused by co-infection with mucormycosis and aspergillus

    Directory of Open Access Journals (Sweden)

    Sharana Mahomed

    2015-01-01

    Full Text Available We report on an unusual case of oro-rhinocerebral disease caused by mucormycosis and aspergillus co-infection in a 54-year-old insulin dependent diabetic patient. Although she was successfully treated with parenteral amphotericin B followed by oral posaconazole, she was left with irreversible blindness of the right eye and multiple cranial nerve palsies.

  10. Cryptic and Rare Aspergillus Species in Brazil: Prevalence in Clinical Samples and In Vitro Susceptibility to Triazoles

    Science.gov (United States)

    Negri, C. E.; Gonçalves, S. S.; Xafranski, H.; Bergamasco, M. D.; Aquino, V. R.; Castro, P. T. O.

    2014-01-01

    Aspergillus spp. are among the most common causes of opportunistic invasive fungal infections in tertiary care hospitals. Little is known about the prevalence and in vitro susceptibility of Aspergillus species in Latin America, because there are few medical centers able to perform accurate identification at the species level. The purpose of this study was to analyze the distribution of cryptic and rare Aspergillus species among clinical samples from 133 patients with suspected aspergillosis admitted in 12 medical centers in Brazil and to analyze the in vitro activity of different antifungal drugs. The identification of Aspergillus species was performed based on a polyphasic approach, as well as sequencing analysis of the internal transcribed spacer (ITS) region, calmodulin, and β-tubulin genes and phylogenetic analysis when necessary. The in vitro susceptibility tests with voriconazole, posaconazole, and itraconazole were performed according to the CLSI M38-A2 document (2008). We demonstrated a high prevalence of cryptic species causing human infection. Only three isolates, representing the species Aspergillus thermomutatus, A. ochraceus, and A. calidoustus, showed less in vitro susceptibility to at least one of the triazoles tested. Accurate identifications of Aspergillus at the species level and with in vitro susceptibility tests are important because some species may present unique resistance patterns against specific antifungal drugs. PMID:25078909

  11. Aspergillus Tracheobronchitis Causing Subtotal Tracheal Stenosis in a Liver Transplant Recipient

    Directory of Open Access Journals (Sweden)

    Sonia Radunz

    2013-01-01

    Full Text Available Invasive aspergillosis is recognized as one of the most significant opportunistic infections after liver transplantation. Diagnosis of invasive aspergillosis in transplant recipients has been proven to be challenging, and optimal approach to the treatment of invasive aspergillosis is still controversial. We here present an unusual case of Aspergillus tracheobronchitis in the setting of liver transplantation. A 47-year-old female patient with persistent dry cough after liver transplantation developed respiratory insufficiency and was readmitted to the intensive care unit 55 days after liver transplantation. A CT scan revealed subtotal tracheal stenosis; bronchoscopy was performed, and extended white mucus coverings causative of the tracheal stenosis were removed. Microbiological assessment isolated Aspergillus fumigatus. The diagnosis was obstructive Aspergillus tracheobronchitis. The patient was started on a treatment of voriconazole 200 mg orally twice daily, adjusted to a trough level of 1–4 mg/L. For further airway management, a tracheal stent had to be implanted. The patient is alive and well 28 months after liver transplantation. Invasive aspergillosis should be considered a possible etiology in liver transplant patients presenting with unspecific symptoms such as persistent dry cough. Optimal strategies for improved and early diagnosis as well as prophylaxis need to be defined.

  12. Chemometric Analysis of the Volatile Compounds Generated by Aspergillus carbonarius Strains Isolated from Grapes and Dried Vine Fruits

    Directory of Open Access Journals (Sweden)

    Zhan Cheng

    2018-02-01

    Full Text Available Ochratoxin A (OTA contamination in grape production is an important problem worldwide. Microbial volatile organic compounds (MVOCs have been demonstrated as useful tools to identify different toxigenic strains. In this study, Aspergillus carbonarius strains were classified into two groups, moderate toxigenic strains (MT and high toxigenic strains (HT, according to OTA-forming ability. The MVOCs were analyzed by GC-MS and the data processing was based on untargeted profiling using XCMS Online software. Orthogonal projection to latent structures discriminant analysis (OPLS-DA was performed using extract ion chromatogram GC-MS datasets. For contrast, quantitative analysis was also performed. Results demonstrated that the performance of the OPLS-DA model of untargeted profiling was better than the quantitative method. Potential markers were successfully discovered by variable importance on projection (VIP and t-test. (E-2-octen-1-ol, octanal, 1-octen-3-one, styrene, limonene, methyl-2-phenylacetate and 3 unknown compounds were selected as potential markers for the MT group. Cuparene, (Z-thujopsene, methyl octanoate and 1 unknown compound were identified as potential markers for the HT groups. Finally, the selected markers were used to construct a supported vector machine classification (SVM-C model to check classification ability. The models showed good performance with the accuracy of cross-validation and test prediction of 87.93% and 92.00%, respectively.

  13. Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

    International Nuclear Information System (INIS)

    Woon, J. S. K.; Murad, A. M. A.; Abu Bakar, F. D.

    2015-01-01

    A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli

  14. Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Woon, J. S. K., E-mail: jameswoon@siswa.ukm.edu.my; Murad, A. M. A., E-mail: munir@ukm.edu.my; Abu Bakar, F. D., E-mail: fabyff@ukm.edu.my [School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor (Malaysia)

    2015-09-25

    A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli.

  15. Production and biochemical characterization of α-glucosidase from Aspergillus niger ITV-01 isolated from sugar cane bagasse.

    Science.gov (United States)

    Del Moral, S; Barradas-Dermitz, D M; Aguilar-Uscanga, M G

    2018-01-01

    Aspergillus niger ITV-01 presents amylolytic activity, identified as α-glucosidase, an enzyme that only produces α-d-glucose from soluble starch and that presents transglucosylase activity on α-d-glucopyranosyl-(1-4)-α-d-glucopyranose (maltose) (200 gL -1 ). Biochemical characterization was performed on A. niger ITV-01 α-glucosidase; its optimum parameters were pH 4.3, temperature 80 °C but stable at 40 °C, with an energy of activation (Ea) 176.25 kJ mol -1 . Using soluble starch as the substrate, K m and V max were 5 mg mL -1 and 1000 U mg -1 , respectively. As α-glucosidase is not a metalloenzyme, calcium and EDTA did not have any effect on its activity. The molecular weight was estimated by SDS-PAGE to be about 75 kDa. It was also active in methanol and ethanol. When ammonium sulfate (AS) and yeast extract (YE) nitrogen sources and calcium effect were evaluated, the greatest activity occurred using YE and calcium, as opposed to AS media where no activity was detected. The results obtained showed that this enzyme has industrial application potential in the processes to produce either ethanol or malto-oligosaccharides from α-d-glucopyranosyl-(1-4)-α-d-glucopyranose (maltose).

  16. Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

    Science.gov (United States)

    Woon, J. S. K.; Murad, A. M. A.; Abu Bakar, F. D.

    2015-09-01

    A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli.

  17. [In vitro activity of ampicillin-ceftriaxone against Enterococcus faecalis isolates recovered from invasive infections].

    Science.gov (United States)

    Burguer Moreira, Noelia; Nastro, Marcela; Vay, Carlos; Famiglietti, Ángela; Rodríguez, Carlos Hernán

    2016-01-01

    In vitro activity of the combination of ampicillin- ceftriaxone against 30 Enterococcus faecalis isolates recovered from invasive infections in patients admitted to Hospital de Clínicas José de San Martin in the city of Buenos Aires was assessed. Ampicillin- ceftriaxone synergies were determined by microdilution in Müeller-Hinton (MH) broth with and without subinhibitory concentrations of ceftriaxone. Synergy was detected in 22/30 isolates. A decrease in both minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) was observed in 14/30 isolates, whereas in 6/30 isolates the decrease was observed in the MIC value and only in the MBC value in the 2 remaining isolates. The bactericidal activity of the combination showed to be higher at low concentrations of ampicillin (ceftriaxone combination and thus, its efficacy was confirmed in the treatment of severe infections by E. faecalis. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  18. Clonal relationship of recent invasive Haemophilus influenzae serotype f isolates from Denmark and the United States

    DEFF Research Database (Denmark)

    Bruun, B; Gahrn-Hansen, B; Westh, H

    2004-01-01

    Surveillance performed after the introduction of general Haemophilus influenzae serotype b (Hib) vaccination in Denmark identified 13 cases of invasive bacteraemic H. influenzae serotype f (Hif) disease in adults over a period of 7 years. Bacteraemic respiratory tract infections accounted for 61...... sequences. Multilocus enzyme electrophoresis typing revealed that recent Danish and American isolates belonged to a single Hif clone, which may be undergoing expansion. The need for accurate serotyping of H. influenzae to enable reliable monitoring for Hib replacement by other capsular types is emphasized....

  19. Diversity in secondary metabolites including mycotoxins from strains of aspergillus section nigri isolated from raw cashew nuts from benin, west africa

    NARCIS (Netherlands)

    Lamboni, Yendouban; Nielsen, Kristian F.; Linnemann, Anita R.; Gezgin, Yué Ksel; Hell, Kerstin; Nout, Rob; Smid, Eddy J.; Tamo, Manuele; Boekel, van M.A.J.S.; Hoof, Jakob Blñsbjerg; Frisvad, Jens Christian

    2016-01-01

    In a previous study, raw cashew kernels were assayed for the fungal contamination focusin on strains belonging to the genus Aspergillus and on aflatoxins producers. These sample showed high contamination with Aspergillus section Nigri species and absence o aflatoxins. To investigate the diversity

  20. Abiotic factors and their interactions influence on the co-production of aflatoxin B(1) and cyclopiazonic acid by Aspergillus flavus isolated from corn.

    Science.gov (United States)

    Astoreca, Andrea; Vaamonde, Graciela; Dalcero, Ana; Marin, Sonia; Ramos, Antonio

    2014-04-01

    The objectives of this study were i) to determine the effects of the interactions of water activity, temperature and incubation time on the co-production of AFB1 and CPA by isolates of Aspergillus flavus with different profile of mycotoxin production and ii) to identify the aW and temperature limiting conditions for the production of both mycotoxins. Fungi used in this study were selected because they belonged to different chemotypes: chemotype I (AFB1+/CPA+), III (AFB1+/CPA-) and IV (AFB1-/CPA+), respectively. Two culture media were used; Czapek yeast agar (CYA) and corn extract agar (CEM), at different incubated temperatures (10-40 °C) and aW levels (0.80-0.98). AFB1 and CPA production were analyzed after 7, 14, 21 and 28 days of incubation. Significant differences were observed with respect to mycotoxin production depending on the media evaluated. The AFB1 production occurred more favorably on CYA while the highest CPA concentrations were recorded on CEM. Within the range of aW evaluated in this study, 0.83 was the limiting level for both toxins production. The optimum conditions for AFB1 production occurred at 0.96 aW and 30 °C after 21 days of incubation, regardless of the media and isolate. Although different amounts of toxins were produced in each medium, the limiting and optimum conditions for their production were similar in both. No differences in the response of the three isolates to the abiotic factors discussed were observed despite belonging to different chemotypes. The determination of the thresholds of mycotoxins co-production, especially in the case of data obtained with the corn extract medium can be useful to avoid the conditions conducive to co-occurrence of these mycotoxins in corn. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

  1. Isolation and characterization of two ammonium permease genes, meaA and mepA, from Aspergillus nidulans.

    Science.gov (United States)

    Monahan, Brendon J; Fraser, James A; Hynes, Michael J; Davis, Meryl A

    2002-02-01

    Ammonium and the analogue methylammonium are taken into the cell by active transport systems which constitute a family of transmembrane proteins that have been identified in fungi, bacteria, plants, and animals. Two genes from Aspergillus nidulans, mepA and meaA, which encode ammonium transporters with different affinities have been characterized. The MepA transporter exhibits the highest affinity for methylammonium (Km, 44.3 microM); in comparison, the Km for MeaA is 3.04 mM. By use of targeted gene replacement strategies, meaA and mepA deletion mutants were created. Deletion of both meaA and mepA resulted in the inability of the strain to grow on ammonium concentrations of less than 10 mM. The single meaA deletion mutant exhibited reduced growth at the same concentrations, whereas the mepA deletion mutant displayed wild-type growth. Interestingly, multiple copies of mepA were found to complement the methylammonium resistance phenotype conferred by the deletion of meaA. The expression profiles for mepA and meaA differed; the mepA transcript was detected only in nitrogen-starved cultures, whereas meaA was expressed under both ammonium-sufficient and nitrogen starvation conditions. Together, these results indicate that MeaA constitutes the major ammonium transport activity and is required for the optimal growth of A. nidulans on ammonium as the sole nitrogen source and that MepA probably functions in scavenging low concentrations of ammonium under nitrogen starvation conditions.

  2. Dissimilatory nitrate reduction by Aspergillus terreus isolated from the seasonal oxygen minimum zone in the Arabian Sea

    Digital Repository Service at National Institute of Oceanography (India)

    Stief, P.; Fuchs-Ocklenburg, S.; Kamp, A.; Manohar, C.S.; Houbraken, J.; Boekhout, T.; deBeer, D.; Stoeck, T.

    , salterns, capybara droppings, lung of pocket mice, corn, cotton plants, milled rice, muesli, and wall paint [39]. The species has been reported from marine and associated habitats, such as mangroves and soft corals, and isolates from these habitats have...

  3. Combination of Voriconazole and Anidulafungin for Treatment of Triazole-Resistant Aspergillus fumigatus in an In Vitro Model of Invasive Pulmonary Aspergillosis

    Science.gov (United States)

    Jeans, Adam R.; Howard, Susan J.; Al-Nakeeb, Zaid; Goodwin, Joanne; Gregson, Lea; Warn, Peter A.

    2012-01-01

    Voriconazole is a first-line agent for the treatment of invasive pulmonary aspergillosis. Isolates with elevated voriconazole MICs are increasingly being seen, and the optimal treatment regimen is not defined. We investigated whether the combination of voriconazole with anidulafungin may be beneficial for the treatment of A. fumigatus strains with elevated voriconazole MICs. We used an in vitro model of the human alveolus to define the exposure-response relationships for a wild-type strain (voriconazole MIC, 0.5 mg/liter) and strains with defined molecular mechanisms of triazole resistance (MICs, 4 to 16 mg/liter). All strains had anidulafungin minimum effective concentrations (MECs) of 0.0078 mg/liter. Exposure-response relationships were estimated using galactomannan as a biomarker. Concentrations of voriconazole and anidulafungin were measured using high-performance liquid chromatography (HPLC). The interaction of voriconazole and anidulafungin was described using the Greco model. Fungal growth was progressively inhibited with higher drug exposures of voriconazole. Strains with elevated voriconazole MICs required proportionally greater voriconazole exposures to achieve a comparable antifungal effect. Galactomannan concentrations were only marginally reduced by anidulafungin monotherapy. An additive effect between voriconazole and anidulafungin was apparent. In conclusion, the addition of anidulafungin does not markedly alter the exposure-response relationship of voriconazole. A rise in serum galactomannan during combination therapy with voriconazole and anidulafungin should be interpreted as treatment failure and not attributed to a paradoxical reaction related to echinocandin treatment. PMID:22825124

  4. Characteristics of Haemophilus influenzae invasive isolates from Portugal following routine childhood vaccination against H. influenzae serotype b (2002-2010).

    Science.gov (United States)

    Bajanca-Lavado, M P; Simões, A S; Betencourt, C R; Sá-Leão, R

    2014-04-01

    We aimed to characterize Haemophilus influenzae invasive isolates recovered in Portugal over a 9-year period (2002-2010) following the inclusion of H. influenzae serotype b (Hib) conjugate vaccination in the National Immunization Program (NIP) in the year 2000 and compare the results with those obtained in a similar study from the pre-vaccination era (1989-2001) previously described by us. As part of a laboratory-based passive surveillance system, 144 invasive isolates obtained in 28 Portuguese hospitals were received at the National Reference Laboratory for Bacterial Respiratory Infections and were characterized. Capsular types and antibiotic susceptibility patterns were determined. The ftsI gene encoding PBP3 was sequenced for β-lactamase-negative ampicillin-resistant (BLNAR) isolates. Genetic relatedness among isolates was examined by multilocus sequencing typing (MLST). Most isolates (77.1%) were non-capsulated, a significant increase compared to the pre-vaccination era (19.0%, p influenzae invasive disease isolates. Most episodes of invasive disease occurring in Portugal are now due to fully susceptible, highly diverse, non-capsulated strains. Given the evolving dynamics of this pathogen and the increase in non-type b capsulated isolates, continuous surveillance is needed.

  5. use of mutagens in the improvement of itaconic acid production by local isolate of aspergillus terreus ref from raw starchy materials and refused banana fruits

    International Nuclear Information System (INIS)

    Khalaf, S.A

    2004-01-01

    Itaconic acid (IA) production by local isolate o/Aspergillus terreus RBF from different acid hydrolyzed starchy materials such as corn starch, potato flour, sweet potato flour and market refused banana fruits extract (MRBFex) was investigated. Both production and yield were highest on corn starch with values of 12.9 g/l and 25.44%, respectively. MRBFex was explored as a good substrate for IA production with yield of '20.42%. The efficiency of IA production by this wild type strain was improved by ultraviolet, chemical and mixed mutagenic treatments. Three high IA yielding mutants, NA 2 , NAUV 2 4 and NASA 5 6 were obtained by gradient plating. One of these mutants (NAUV 2 4) was capable of producing about twice the yield of IA as the parent strain from acid hydrolyzed corn starch and/or MRBFex. Higher than 30 g/l of IA was produced by the mutant strain NAUV 2 4 in flask fermentation from medium containing of 120 g/l (as sugar) of MRBFex

  6. Response surface methodology for production, characterization and application of solvent, salt and alkali-tolerant alkaline protease from isolated fungal strain Aspergillus niger WA 2017.

    Science.gov (United States)

    Abdel Wahab, Walaa A; Ahmed, Samia A

    2018-04-17

    Isolated strain Aspergillus niger WA 2017 was selected as potential protease producer and was identified on the basis of 18S rDNA gene homology. Optimization of protease production conditions was performed using statistical methodology. The most significant factors were identified by Plackett-Burman design (PB) and were optimized by central composite design (CCD). The enzyme production was increased by 3.6-fold with statistically optimized medium when compared to the basal medium. Based on the protease activity, 25-50% ethanol fraction exhibited the highest specific activity. The partially purified enzyme showed its highest activity (4.7-fold) after 10 min incubation at pH 10.0 and 60 °C. The enzyme was stable over a wide range of pH (7-11) and salt concentration (up to 20%). Kinetic parameters Michaelis constant (K m ) and maximum velocity (V max ) were calculated at varying casein concentrations. Additionally, thermal stability of the enzyme was substantially improved by NaCl. The enzyme showed excellent stability and compatibility in presence of organic solvents and detergents retaining 115.3 and 114.5% of its activity in presence of ethanol and Tide, respectively at 40 °C for 1 h. The results revealed that the produced enzyme was able to recover silver from used X-ray film under optimized condition using statistical methodology (CCD). Copyright © 2017. Published by Elsevier B.V.

  7. OPTIMIZATION OF MILK-CLOTTING PROTEASE PRODUCTION BY A LOCAL ISOLATE OF ASPERGILLUS NIGER FFB1 IN SOLID-STATE FERMENTATION

    Directory of Open Access Journals (Sweden)

    Souhila Bensmail

    2015-04-01

    Full Text Available The need to surmount the limitation of obtaining rennin, has been actively pushed researches to find new substitutes that present high milk-clotting activity which enables the production of high yields of cheese. In this study, the production of extracellular milk-clotting protease by locally isolated fungal specie, Aspergillus niger FFB1 under solid-state fermentation (SSF using cheep agro-industrial byproduct (wheat bran was optimized. The effects of several physicochemical and environmental factors were investigated to select the optimal conditions that ensure the best milk-clotting activity by application of "One-factor-at-a-time" method. A trial of cheese production using the crude extract was also carried out. The maximum enzyme activity (830 SU/g bran with a ratio MCA/PA of 4.25 was obtained under the optimum conditions of temperature (30°C, spores concentration (106 spores/mL, incubation time (72 hours, and moisture content of solid substrate (39.2% adjusted suitably with mineral solution (Czapek-Dox of pH 4.

  8. Comparison of Two Molecular Assays for Detection and Characterization of Aspergillus fumigatus Triazole Resistance and Cyp51A Mutations in Clinical Isolates and Primary Clinical Samples of Immunocompromised Patients

    OpenAIRE

    Patricia Postina; Julian Skladny; Tobias Boch; Oliver A. Cornely; Oliver A. Cornely; Axel Hamprecht; Peter-Michael Rath; Jörg Steinmann; Oliver Bader; Thomas Miethke; Anne Dietz; Natalia Merker; Wolf-Karsten Hofmann; Dieter Buchheidt; Birgit Spiess

    2018-01-01

    In hematological patients, the incidence of invasive aspergillosis (IA) caused by azole resistant Aspergillus fumigatus (ARAf) is rising. As the diagnosis of IA is rarely based on positive culture in this group of patients, molecular detection of resistance mutations directly from clinical samples is crucial. In addition to the in-house azole resistance ARAf polymerase chain reaction (PCR) assays detecting the frequent mutation combinations TR34/L98H, TR46/Y121F/T289A, and M220 in the Aspergi...

  9. Isolation and Characterization of “Terrein” an Antimicrobial and Antitumor Compound from Endophytic Fungus Aspergillus terreus (JAS-2 Associated from Achyranthus aspera Varanasi, India

    Directory of Open Access Journals (Sweden)

    Jyoti Goutam

    2017-07-01

    Full Text Available The present study aimed at characterizing biological potentials of endophyte Aspergillus terreus JAS-2 isolated from Achyranthus aspera. Crude extracted from endophytic fungus JAS-2 was purified and chemically characterized by chromatographic and spectroscopic studies respectively. Spectral assignment of NMR (nuclear magnetic resonance data, 1H proton and 13C carbon analysis along with FTIR data elucidated the structure of compound as 4,5-Dihydroxy-3-(1-propenyl-2-cyclopenten-1-one. After purification and identification a set of experiment was conducted to explore efficacy of compound. Results revealed that on accessing the antifungal activity of compound, growth diameter of tested phytopathogenic fungi was reduced to 50% at higher concentration taken (10 μgμl−1. Compound exhibited in-vitro bacterial cell inhibition at 20 μgml−1 concentration along with moderate antioxidant behavior. Evaluation of anticancer activity against human lung cancer cell line (A-549 exhibited its IC50 value to be 121.9 ± 4.821 μgml−1. Further cell cycle phase distribution were analyzed on the basis of DNA content and evaluated by FACS (Fluorescence Activated Cell Sorting and it was revealed that at 150 μgml−1 of compound maximum cells were found in sub G1 phase which represents apoptotic dead cells. Terrein (4, 5-Dihydroxy-3-(1-propenyl-2-cyclopenten-1-one a multi-potential was isolated from endophytic fungus JAS-2, from well recognized medicinal herb A. aspera. To best of our knowledge, this is the first report of “Terrein” from endophytic derived fungus. This compound had also exhibited anticancer and antifungal activity against human lung cancer cell line A-549 and Bipolaris sorokiniana respectively which is causal organism of many plants disease. Hence endophytes are serving as alternative sources of drug molecules.

  10. Aspergillus infections of the lung

    International Nuclear Information System (INIS)

    Nicolas, V.; Hottenrott, K.; Hossfeld, D.K.

    1989-01-01

    In this article the three different manifestations of pulmonary aspergillosis are compared with each other: Bronchopulmonary aspergillosis, aspergilloma and invasive pulmonary aspergillosis, and their different clinical and radiological manifestations. In particular, invasive pulmonary aspergillosis - being a severe complication in immuno-suppressed patients whose defenses are weak - often presents diagnostic problems due to its initially mostly unspecific clinical and radiological signs. The late signs of a pulmonary aspergillus infection, such as formation of a cavern or the manifestation of the 'air crescent' sign, however, are seen only when the patient is just beginning to convalesce. (orig.) [de

  11. Sentinel Lymph Node Biopsy and Isolated Tumor Cells in Invasive Lobular Versus Ductal Breast Cancer.

    Science.gov (United States)

    Truin, Wilfred; Roumen, Rudi M; Siesling, Sabine; van der Heiden-van der Loo, Margriet; Lobbezoo, Dorien J; Tjan-Heijnen, Vivianne C; Voogd, Adri C

    2016-08-01

    Sentinel lymph node (SLN) biopsy is the standard of care for axillary staging in invasive breast cancer. The introduction of SLN biopsy with an extensive pathology examination, in addition to the introduction of the 2002 TNM classification, led to different axillary classification outcomes. We evaluated the effect of axillary staging procedures and subsequent axillary nodal status in patients with invasive lobular carcinoma (ILC) versus invasive ductal carcinoma (IDC) from 1998 to 2013. The use of SLN biopsy and the nodal status distribution were analyzed in patients with stage T1-T2 ILC and IDC. Logistic regression analysis was performed to determine the independent effect of histologic type on the probability of the presence of isolated tumor cells (ITCs), micrometastases, and macrometastases. A total of 89,971 women were diagnosed, 10,146 with ILC (11%) and 79,825 with IDC (89%). The patients who had undergone SLN biopsy were more frequently diagnosed with ITCs than were those who had undergone axillary lymph node dissection only (odds ratio, 8.8; 95% confidence interval, 7.0-11.2). In 2013, the proportion of patients with ITCs in the axillary nodes was 8% in those with ILC and 4.4% in those with IDC. Patients with ILC were significantly more likely to have ITCs in their axillary lymph nodes than were patients with IDC (odds ratio, 1.8; 95% confidence interval, 1.6-2.0). With the introduction of SLN biopsy and the renewed 2002 TNM classification, patients with ILC have been more frequently diagnosed with ITCs than have patients with IDC. The clinical consequence of this finding must be established from further research. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Heterogeneity in the Infection Biology of Campylobacter jejuni Isolates in Three Infection Models Reveals an Invasive and Virulent Phenotype in a ST21 Isolate from Poultry.

    Directory of Open Access Journals (Sweden)

    Suzanne Humphrey

    Full Text Available Although Campylobacter is the leading cause of bacterial foodborne gastroenteritis in the world and the importance of poultry as a source of infection is well understood we know relatively little about its infection biology in the broiler chicken. Much of what we know about the biology of Campylobacter jejuni is based on infection of inbred or SPF laboratory lines of chickens with a small number of isolates used in most laboratory studies. Recently we have shown that both the host response and microbial ecology of C. jejuni in the broiler chicken varies with both the host-type and significantly between C. jejuni isolates. Here we describe heterogeneity in infection within a panel of C. jejuni isolates in two broiler chicken breeds, human intestinal epithelial cells and the Galleria insect model of virulence. All C. jejuni isolates colonised the chicken caeca, though colonisation of other parts of the gastrointestinal tract varied between isolates. Extra-intestinal spread to the liver varied between isolates and bird breed but a poultry isolate 13126 (sequence type 21 showed the greatest levels of extra-intestinal spread to the liver in both broiler breeds with over 70% of birds of the fast growing breed and 50% of the slower growing breed having C. jejuni in their livers. Crucially 13126 is significantly more invasive than other isolates in human intestinal epithelial cells and gave the highest mortality in the Galleria infection model. Taken together our findings suggest that not only is there considerable heterogeneity in the infection biology of C. jejuni in avian, mammalian and alternative models, but that some isolates have an invasive and virulent phenotype. Isolates with an invasive phenotype would pose a significant risk and increased difficulty in control in chicken production and coupled with the virulent phenotype seen in 13126 could be an increased risk to public health.

  13. Antifungal susceptibility of invasive yeast isolates in Italy: the GISIA3 study in critically ill patients

    Directory of Open Access Journals (Sweden)

    Mussap Michele

    2011-05-01

    Full Text Available Abstract Background Yeasts are a common cause of invasive fungal infections in critically ill patients. Antifungal susceptibility testing results of clinically significant fungal strains are of interest to physicians, enabling them to adopt appropriate strategies for empiric and prophylactic therapies. We investigated the antifungal susceptibility of yeasts isolated over a 2-year period from hospitalised patients with invasive yeast infections. Methods 638 yeasts were isolated from the blood, central venous catheters and sterile fluids of 578 patients on general and surgical intensive care units and surgical wards. Etest strips and Sensititre panels were used to test the susceptibility of the isolates to amphotericin B, anidulafungin, caspofungin, fluconazole, itraconazole, posaconazole and voriconazole in 13 laboratories centres (LC and two co-ordinating centres (CC. The Clinical and Laboratory Standards Institute (CLSI reference broth microdilution method was used at the CCs for comparison. Results Etest and Sensititre (LC/CC MIC90 values were, respectively: amphotericin B 0.5/0.38, 1/1 mg/L; anidulafungin 2/1.5 and 1/1 mg/L; caspofungin 1/0.75 and 0.5/0.5 mg/L; fluconazole 12/8 and 16/16 mg/L; itraconazole 1/1.5, 0.5/0.5 mg/L; posaconazole 0.5 mg/L and voriconazole 0.25 mg/L for all. The overall MIC90 values were influenced by the reduced susceptibility of Candida parapsilosis isolates to echinocandins and a reduced or lack of susceptibility of Candida glabrata and Candida krusei to azoles, in particular fluconazole and itraconazole. Comparison of the LC and CC results showed good Essential Agreement (90.3% for Etest and 92.9% for Sensititre, and even higher Categorical Agreement (93.9% for Etest and 96% for Sensititre; differences were observed according to the species, method, and antifungal drug. No cross-resistance between echinocandins and triazoles was detected. Conclusions Our data confirm the different antifungal susceptibility

  14. Antimicrobial activities of endophytic fungi obtained from the arid zone invasive plant Opuntia dillenii and the isolation of equisetin, from endophytic Fusarium sp.

    Science.gov (United States)

    Ratnaweera, Pamoda B; de Silva, E Dilip; Williams, David E; Andersen, Raymond J

    2015-07-10

    Opuntia dillenii is an invasive plant well established in the harsh South-Eastern arid zone of Sri Lanka. Evidence suggests it is likely that the endophytic fungal populations of O. dillenii assist the host in overcoming biotic and abiotic stress by producing biologically active metabolites. With this in mind there is potential to discover novel natural products with useful biological activities from this hitherto poorly investigated source. Consequently, an investigation of the antimicrobial activities of the endophytes of O. dillenii, that occupies a unique ecological niche, may well provide useful leads in the discovery of new pharmaceuticals. Endophytic fungi were isolated from the surface sterilized cladodes and flowers of O. dillenii using several nutrient media and the antimicrobial activities were evaluated against three Gram-positive and two Gram-negative bacteria and Candida albicans. The two most bioactive fungi were identified by colony morphology and DNA sequencing. The secondary metabolite of the endophyte Fusarium sp. exhibiting the best activity was isolated via bioassay guided chromatography. The chemical structure was elucidated from the ESIMS and NMR spectroscopic data obtained for the active metabolite. The minimum inhibitory concentrations (MICs) of the active compound were determined. Eight endophytic fungi were isolated from O. dillenii and all except one showed antibacterial activities against at least one of the test bacteria. All extracts were inactive against C. albicans. The most bioactive fungus was identified as Fusarium sp. and the second most active as Aspergillus niger. The structure of the major antibacterial compound of the Fusarium sp. was shown to be the tetramic acid derivative, equisetin. The MIC's for equisetin were 8 μg mL(-1) against Bacillus subtilis, 16 μg mL(-1) against Staphylococcus aureus and Methicillin Resistant Staphylococcus aureus (MRSA). O. dillenii, harbors several endophytic fungi capable of producing

  15. Biosolubilization of poorly soluble rock phosphates by Aspergillus tubingensis and Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Reddy, M.S.; Kumar, S.; Babita, K. [Thapar Institute of Engineering and Technology, Patiala (India). School of Biotechnology; Reddy, M.S. [Auburn University, AL (United States). Department of Entomology and Plant Pathology

    2002-09-01

    Three isolates of Aspergillus tubingensis and two isolates of Aspergillus niger isolated from rhizospheric soils were tested on solubilization of different rock phosphates. All the isolates of Aspergillus were capable of solubilizing all the natural rock phosphates. A. tubingensis (AT1) showed maximum percent solubilization in all the rock phosphates tested in this study when compared to other isolates. This isolate also showed highest phosphorus (P) solubilization when grown in the presence of 2% of rock phosphate. A. tubingensis (AT1) seems to be more efficient in solubilization of rock phosphates compared to other isolates reported elsewhere. This is the first report of rock phosphate solubilization by A. tubingensis and might provide an efficient large scale biosolubilization of rock phosphates intended for P fertilizer. (author)

  16. Taxonomic Characterization and Secondary Metabolite Profiling of Aspergillus Section Aspergillus Contaminating Feeds and Feedstuffs

    Directory of Open Access Journals (Sweden)

    Mariana Greco

    2015-09-01

    Full Text Available Xerophilic fungal species of the genus Aspergillus are economically highly relevant due to their ability to grow on low water activity substrates causing spoilage of stored goods and animal feeds. These fungi can synthesize a variety of secondary metabolites, many of which show animal toxicity, creating a health risk for food production animals and to humans as final consumers, respectively. Animal feeds used for rabbit, chinchilla and rainbow trout production in Argentina were analysed for the presence of xerophilic Aspergillus section Aspergillus species. High isolation frequencies (>60% were detected in all the studied rabbit and chinchilla feeds, while the rainbow trout feeds showed lower fungal charge (25%. These section Aspergillus contaminations comprised predominantly five taxa. Twenty isolates were subjected to taxonomic characterization using both ascospore SEM micromorphology and two independent DNA loci sequencing. The secondary metabolite profiles of the isolates were determined qualitatively by HPLC-MS. All the isolates produced neoechinulin A, 17 isolates were positive for cladosporin and echinulin, and 18 were positive for neoechinulin B. Physcion and preechinulin were detected in a minor proportion of the isolates. This is the first report describing the detailed species composition and the secondary metabolite profiles of Aspergillus section Aspergillus contaminating animal feeds.

  17. Draft Genome Sequence of an Invasive Multidrug-Resistant Strain, Pseudomonas aeruginosa BK1, Isolated from a Keratitis Patient

    KAUST Repository

    Jeganathan, Lakshmi Priya

    2014-03-27

    Pseudomonas aeruginosa infections are difficult to treat due to the presence of a multitude of virulence factors and antibiotic resistance. Here, we report the draft genome sequence of P. aeruginosa BK1, an invasive and multidrug-resistant strain, isolated from a bacterial keratitis patient in southern India.

  18. Characterisation of Invasive Streptococcus pneumoniae Isolated from Cambodian Children between 2007 - 2012.

    Science.gov (United States)

    Moore, Catrin E; Giess, Adam; Soeng, Sona; Sar, Poda; Kumar, Varun; Nhoung, Pheakdey; Bousfield, Rachel; Turner, Paul; Stoesser, Nicole; Day, Nicholas P J; Parry, Christopher M

    2016-01-01

    The 13-valent pneumococcal vaccine (PCV13) was introduced in Cambodia in January 2015. There are limited data concerning the common serotypes causing invasive pneumococcal disease (IPD). Knowledge of the circulating pneumococcal serotypes is important to monitor epidemiological changes before and after vaccine implementation. All episodes of IPD defined by the isolation of Streptococcus pneumoniae from blood, cerebrospinal fluid or other sterile site in Cambodian children admitted to the Angkor Hospital for Children in Siem Reap, Northwestern Cambodia, between 1st January 2007 and 1st July 2012 were retrospectively studied. Streptococcus pneumoniae isolates that could be retrieved underwent phenotypic typing and whole genome sequencing. There were 90 Cambodian children hospitalized with IPD with a median (IQR) age of 2.3 years (0.9-6.2). The case fatality was 15.6% (95% CI 8-23). Of 50 Streptococcus pneumoniae isolates available for further testing, 46% were penicillin non-susceptible and 8% were ceftriaxone non-susceptible, 78% were cotrimoxazole resistant, 30% were erythromycin resistant and 30% chloramphenicol resistant. There were no significant changes in resistance levels over the five-year period. The most common serotypes were 1 (11/50; 22%), 23F (8/50; 16%), 14 (6/50; 12%), 5 (5/50; 10%) and 19A (3/50; 6%). Coverage by PCV7, PCV10 and PCV13 was 44%, 76% and 92% respectively. We identified novel multilocus sequence types and resistotypes using whole genome sequencing. This study suggests IPD is an important disease in Cambodian children and can have a significant mortality. PCV13 coverage of the serotypes determined in studied strains was high and consistent with another recent study. The phenotypic resistance patterns observed were similar to other regional studies. The use of whole genome sequencing in the present study provides additional typing and resistance information together with the description of novel sequence types and resistotypes.

  19. Characterisation of Invasive Streptococcus pneumoniae Isolated from Cambodian Children between 2007 - 2012.

    Directory of Open Access Journals (Sweden)

    Catrin E Moore

    Full Text Available The 13-valent pneumococcal vaccine (PCV13 was introduced in Cambodia in January 2015. There are limited data concerning the common serotypes causing invasive pneumococcal disease (IPD. Knowledge of the circulating pneumococcal serotypes is important to monitor epidemiological changes before and after vaccine implementation.All episodes of IPD defined by the isolation of Streptococcus pneumoniae from blood, cerebrospinal fluid or other sterile site in Cambodian children admitted to the Angkor Hospital for Children in Siem Reap, Northwestern Cambodia, between 1st January 2007 and 1st July 2012 were retrospectively studied. Streptococcus pneumoniae isolates that could be retrieved underwent phenotypic typing and whole genome sequencing.There were 90 Cambodian children hospitalized with IPD with a median (IQR age of 2.3 years (0.9-6.2. The case fatality was 15.6% (95% CI 8-23. Of 50 Streptococcus pneumoniae isolates available for further testing, 46% were penicillin non-susceptible and 8% were ceftriaxone non-susceptible, 78% were cotrimoxazole resistant, 30% were erythromycin resistant and 30% chloramphenicol resistant. There were no significant changes in resistance levels over the five-year period. The most common serotypes were 1 (11/50; 22%, 23F (8/50; 16%, 14 (6/50; 12%, 5 (5/50; 10% and 19A (3/50; 6%. Coverage by PCV7, PCV10 and PCV13 was 44%, 76% and 92% respectively. We identified novel multilocus sequence types and resistotypes using whole genome sequencing.This study suggests IPD is an important disease in Cambodian children and can have a significant mortality. PCV13 coverage of the serotypes determined in studied strains was high and consistent with another recent study. The phenotypic resistance patterns observed were similar to other regional studies. The use of whole genome sequencing in the present study provides additional typing and resistance information together with the description of novel sequence types and resistotypes.

  20. Characterisation of Invasive Streptococcus pneumoniae Isolated from Cambodian Children between 2007 – 2012

    Science.gov (United States)

    Giess, Adam; Soeng, Sona; Sar, Poda; Kumar, Varun; Nhoung, Pheakdey; Bousfield, Rachel; Turner, Paul; Stoesser, Nicole; Day, Nicholas P. J.; Parry, Christopher M.

    2016-01-01

    Background The 13-valent pneumococcal vaccine (PCV13) was introduced in Cambodia in January 2015. There are limited data concerning the common serotypes causing invasive pneumococcal disease (IPD). Knowledge of the circulating pneumococcal serotypes is important to monitor epidemiological changes before and after vaccine implementation. Methods All episodes of IPD defined by the isolation of Streptococcus pneumoniae from blood, cerebrospinal fluid or other sterile site in Cambodian children admitted to the Angkor Hospital for Children in Siem Reap, Northwestern Cambodia, between 1st January 2007 and 1st July 2012 were retrospectively studied. Streptococcus pneumoniae isolates that could be retrieved underwent phenotypic typing and whole genome sequencing. Results There were 90 Cambodian children hospitalized with IPD with a median (IQR) age of 2.3 years (0.9–6.2). The case fatality was 15.6% (95% CI 8–23). Of 50 Streptococcus pneumoniae isolates available for further testing, 46% were penicillin non-susceptible and 8% were ceftriaxone non-susceptible, 78% were cotrimoxazole resistant, 30% were erythromycin resistant and 30% chloramphenicol resistant. There were no significant changes in resistance levels over the five-year period. The most common serotypes were 1 (11/50; 22%), 23F (8/50; 16%), 14 (6/50; 12%), 5 (5/50; 10%) and 19A (3/50; 6%). Coverage by PCV7, PCV10 and PCV13 was 44%, 76% and 92% respectively. We identified novel multilocus sequence types and resistotypes using whole genome sequencing. Conclusions This study suggests IPD is an important disease in Cambodian children and can have a significant mortality. PCV13 coverage of the serotypes determined in studied strains was high and consistent with another recent study. The phenotypic resistance patterns observed were similar to other regional studies. The use of whole genome sequencing in the present study provides additional typing and resistance information together with the description of novel

  1. Clonal replacement and expansion among invasive meningococcal isolates of serogroup W in France.

    Science.gov (United States)

    Hong, Eva; Barret, Anne-Sophie; Terrade, Aude; Denizon, Mélanie; Antona, Denise; Aouiti-Trabelsi, Myriam; Deghmane, Ala-Eddine; Parent du Châtelet, Isabelle; Levy-Bruhl, Daniel; Taha, Muhamed-Kheir

    2018-02-01

    Neisseria meningitidis group W (NmW) belonging to the clonal complex ST-11 (NmW/cc11) spread in Europe and in France in 2000 and declined thereafter. In France, invasive meningococcal disease (IMD) due to NmW increased again in 2012 and thereafter since 2015. Several sub-lineages of NmW/cc11 are circulating worldwide with successive epidemic waves. We aimed to describe recent epidemiological trends of NmW in France and to explore the microbiological and epidemiological characteristics associated with different NmW/cc11 sub-lineages. The epidemiology of NmW was described based on data collected through mandatory notification of IMD and strain typing data for culture-confirmed and PCR-confirmed cases for the period 2000-2016. All culture-confirmed cases due to NmW from the period 2010-2016 were characterised by whole genome sequencing (WGS). A detailed epidemiological analysis was performed for culture-confirmed cases on the basis of WGS data. During the period 2010-2016, genotyping was obtained for 148 cases including all the 132 culture-confirmed cases, among which 127 were matched with epidemiological data, and 16 PCR-confirmed cases (out of a total of 47 PCR-confirmed cases). An increase in IMD was observed in 2012 and was linked to isolates belonging to the "Anglo-French-Hajj" sub-lineage. These isolates have decreased significantly since 2013 and have been replaced by NmW/cc11 isolates related to the "South American - UK" sub-lineage which caused a marked increase in the number of cases of NmW in 2016. In this sub-lineage, the "original UK strain" was first detected in 2012 and increased thereafter, followed by the recently described "UK 2013-strain". Isolates related to the "South American-UK" sub-lineage represented 45% of all NmW cultured isolates from the whole period 2010-2016 but were the most frequent isolates in 2016, representing 76% of the total NmW typed isolates and 94% of the typed NmW/cc11 isolates. A changing pattern in the epidemiology of Nm

  2. [Utility of Aspergillus-LFD: first experience in Chile].

    Science.gov (United States)

    Alvarez, Eduardo

    2015-02-01

    The diagnosis of invasive aspergillosis remains a challenge. Detection of galactomannan in serum and bronchoalveolar lavage is a useful tool; however due to methodological and economic reasons, the test frequencies of galactomannan assays vary from daily to weekly, which constitute a risk to the patient. In this study, we aimed to evaluate and correlate the performance of the new kit Aspergillus-LFD with the GM-EIA. Aspergillus-LFD kit represents a fast, economical and simple test; showed a good performance and excellent correlation with GM-EIA kit. Given the above, the Aspergillus-LFD is emerging as an alternative to consider in the early diagnosis of invasive aspergillosis.

  3. Differential expression of hemoglobin receptor, HmbR, between carriage and invasive isolates of Neisseria meningitidis contributes to virulence: lessons from a clonal outbreak.

    Science.gov (United States)

    Sevestre, Julien; Diene, Seydina M; Aouiti-Trabelsi, Myriam; Deghmane, Ala-Eddine; Tournier, Isabelle; François, Patrice; Caron, François; Taha, Muhamed-Kheir

    2018-04-11

    Carriage and invasion balance in the pathogenesis of Neisseria meningitidis was analyzed during a recent clonal outbreak of meningococcal B in Normandy, France, that offered the opportunity to compare six isolates undistinguable by conventional typing (B:P1.7,16:F3-3/ST-32) isolated from invasive disease or pharyngeal asymptomatic carriage. Data from animal model (transgenic mice rendered susceptible to N. meningitidis infection) showed an absence of virulence for two non-capsulated carriage isolates, an intermediate virulence for two capsulated carriage isolates and a marked virulence for two capsulated invasive isolates. This differential pathogenesis well correlated with whole genome sequencing analysis that clustered together both isolates of each group together, forming their own arm within the Norman cluster. Gene-by-gene analysis specified that genes involved in iron acquisition were among the elements differentially represented in cluster of invasive isolates compared to cluster of capsulated carriage isolates. The hemoglobin receptor encoding gene hmbR was in an ON-phase in the capsulated invasive isolates while carriage capsulated isolates were in an OFF-phase. An ON-phase variant of a capsulated carriage isolate showed enhanced virulence. These data underline the role of phase variation (ON/OFF) of HmbR in the balance between disease isolates/carriage isolates.

  4. Isolation, identification and toxigenic potential of ochratoxin A-producing Aspergillus species from coffee beans grown in two regions of Thailand

    DEFF Research Database (Denmark)

    Noonim, P.; Mahakarnchanakul, W.; Nielsen, Kristian Fog

    2008-01-01

    In 2006 and 2007, 32 Thai dried coffee bean samples (Coffea arabica) from two growing sites of Chiang Mai Province, and 32 Thai dried coffee bean samples (Coffea canephora var. robusta) from two growing sites of Chumphon Province, Thailand, were collected and assessed for the distribution of fungi...... with the potential to produce ochratoxin A (OTA). The overall percentage of fungal contamination in coffee was 98% and reduced to 60% after surface disinfection. There were remarkable ecological differences in the composition of ochratoxigenic species present in these two regions. Arabica coffee bean samples from...... the North had an average of 78% incidence of colonization with Aspergillus of section Circumdati with Aspergillus westerdijkiae and A. melleus as the predominant species. Aspergillus spp. of section Nigri were found in 75% of the samples whereas A. ochraceus was not detected. Robusta coffee beans from...

  5. Aspergillus PCR: one step closer to standardization.

    NARCIS (Netherlands)

    White, P.L.; Bretagne, S.; Klingspor, L.; Melchers, W.J.G.; McCulloch, E.; Schulz, B.; Finnstrom, N.; Mengoli, C.; Barnes, R.A.; Donnelly, J.P.; Loeffler, J.

    2010-01-01

    PCR has been used as an aid in the diagnosis of invasive aspergillosis for almost 2 decades. A lack of standardization has limited both its acceptance as a diagnostic tool and multicenter clinical evaluations, preventing its inclusion in disease-defining criteria. In 2006, the European Aspergillus

  6. Chronic necrotising pneumonia caused by Aspergillus niger.

    Science.gov (United States)

    Wiggins, J; Clark, T J; Corrin, B

    1989-01-01

    A woman with asthma developed chronic necrotising semi-invasive pneumonia due to mixed Aspergillus niger and Candida albicans infection; though not severely immunosuppressed, she may have been predisposed by long term oral corticosteroid and recurrent oral antibiotic treatment. The diagnosis should be considered in patients with chronic airflow limitation who develop cavitating pneumonia. Images PMID:2763249

  7. Use of prophylactic voriconazole for three months after lung transplantation does not reduce infection with Aspergillus

    DEFF Research Database (Denmark)

    Tofte, Nete; Jensen, Claus; Tvede, Michael

    2012-01-01

    This was a retrospective study analyzing the mortality and incidence of Aspergillus infection and invasive disease, comparing patients given voriconazole for 3 months following transplantation to patients not given prophylaxis.......This was a retrospective study analyzing the mortality and incidence of Aspergillus infection and invasive disease, comparing patients given voriconazole for 3 months following transplantation to patients not given prophylaxis....

  8. Cell invasion of poultry-associated Salmonella enterica serovar Enteritidis isolates is associated with pathogenicity, motility and proteins secreted by the type III secretion system.

    Science.gov (United States)

    Shah, Devendra H; Zhou, Xiaohui; Addwebi, Tarek; Davis, Margaret A; Orfe, Lisa; Call, Douglas R; Guard, Jean; Besser, Thomas E

    2011-05-01

    Salmonella enterica serovar Enteritidis (S. Enteritidis) is a major cause of food-borne gastroenteritis in humans worldwide. Poultry and poultry products are considered the major vehicles of transmission to humans. Using cell invasiveness as a surrogate marker for pathogenicity, we tested the invasiveness of 53 poultry-associated isolates of S. Enteritidis in a well-differentiated intestinal epithelial cell model (Caco-2). The method allowed classification of the isolates into low (n = 7), medium (n = 18) and high (n = 30) invasiveness categories. Cell invasiveness of the isolates did not correlate with the presence of the virulence-associated gene spvB or the ability of the isolates to form biofilms. Testing of representative isolates with high and low invasiveness in a mouse model revealed that the former were more invasive in vivo and caused more and earlier mortalities, whereas the latter were significantly less invasive in vivo, causing few or no mortalities. Further characterization of representative isolates with low and high invasiveness showed that most of the isolates with low invasiveness had impaired motility and impaired secretion of either flagella-associated proteins (FlgK, FljB and FlgL) or type III secretion system (TTSS)-secreted proteins (SipA and SipD) encoded on Salmonella pathogenicity island-1. In addition, isolates with low invasiveness had impaired ability to invade and/or survive within chicken macrophages. These data suggest that not all isolates of S. Enteritidis recovered from poultry may be equally pathogenic, and that the pathogenicity of S. Enteritidis isolates is associated, in part, with both motility and secretion of TTSS effector proteins.

  9. Effect of the essential oils from Melaleuca alternifolia, Melaleuca quinquenervia and Backhousia citriodora on the synthesis of ochratoxin A by Aspergillus niger and Aspergillus carbonarius isolated from tropical wine grapes.

    Science.gov (United States)

    de Andrade Santiago, Juliana; Cardoso, Maria das Graças; Batista, Luis Roberto; Santiago, Wilder Douglas; Passamani, Fabiana Reinis Franca; Rodrigues, Leonardo Milani Avelar; Nelson, David Lee

    2018-01-01

    The influence of essential oils (EOs) extracted from the leaves of Melaleuca alternifolia , Melaleuca quinquenervia and Backhousia citriodora on ochratoxin A (OTA) synthesis by fungi was studied. The extraction of EOs was performed by hydrodistillation (Clevenger apparatus) over a 2-h period and subsequently analyzed by GC-MS and GC-FID. The toxigenic activity of the essential oils (31.25; 15.62 and 7.81 µg mL -1 ) was evaluated by inhibiting the production of OTA by Aspergillus niger and Aspergillus carbonarius in Czapek agar medium culture. The quantification of the toxin was performed by HPLC. The production of OTA was dependent on the fungal species, incubation temperature (15 and 25 °C) and the presence of the essential oils. In tests carried out at 15 °C, the oils caused a reduction in OTA synthesis that ranged from 57.60 to 76.93% and from 54.78 to 98.68% for the fungal species A. carbonarius and A. niger , respectively. At 25 °C, reductions ranged from the 38.66 to 75.93% and from 17.94 to 71.79% for the respective fungi. The study concluded that natural products could be potential biocontrol agents against OTA contamination in food.

  10. Characterization of freshly retrieved preantral follicles using a low-invasive, mechanical isolation method extended to different ruminant species.

    Science.gov (United States)

    Langbeen, A; Jorssen, E P A; Fransen, E; Rodriguez, A P A; García, M Chong; Leroy, J L M R; Bols, P E J

    2015-10-01

    Due to the increased interest in preantral follicular physiology, non-invasive retrieval and morphological classification are crucial. Therefore, this study aimed: (1) to standardize a minimally invasive isolation protocol, applicable to three ruminant species; (2) to morphologically classify preantral follicles upon retrieval; and (3) to describe morphological features of freshly retrieved follicles compared with follicle characteristics using invasive methods. Bovine, caprine and ovine ovarian cortex strips were retrieved from slaughterhouse ovaries and dispersed. This suspension was filtered, centrifuged, re-suspended and transferred to a Petri dish, to which 0.025 mg/ml neutral red (NR) was added to assess the viability of the isolated follicles. Between 59 and 191 follicles per follicle class and per species were collected and classified by light microscopy, based on follicular cell morphology. Subsequently, follicle diameters were measured. The proposed isolation protocol was applicable to all three species and showed a significant, expected increase in diameter with developmental stage. With an average diameter of 37 ± 5 μm for primordial follicles, 47 ± 6.3 μm for primary follicles and 67.1 ± 13.1 μm for secondary follicles, no significant difference in diameter among the three species was observed. Bovine, caprine and ovine follicles (63, 59 and 50% respectively) were graded as viable upon retrieval. Using the same morphological characteristics as determined by invasive techniques [e.g. haematoxylin-eosin (HE) sections], cumulus cell morphology and follicle diameter could be used routinely to classify freshly retrieved follicles. Finally, we applied a mechanical, minimally invasive, follicle isolation protocol and extended it to three ruminant species, yielding viable preantral follicles without compromising further in vitro processing and allowing routine follicle characterization upon retrieval.

  11. The frequency of genes encoding three putative group B streptococcal virulence factors among invasive and colonizing isolates

    Directory of Open Access Journals (Sweden)

    Borchardt Stephanie M

    2006-07-01

    Full Text Available Abstract Background Group B Streptococcus (GBS causes severe infections in very young infants and invasive disease in pregnant women and adults with underlying medical conditions. GBS pathogenicity varies between and within serotypes, with considerable variation in genetic content between strains. Three proteins, Rib encoded by rib, and alpha and beta C proteins encoded by bca and bac, respectively, have been suggested as potential vaccine candidates for GBS. It is not known, however, whether these genes occur more frequently in invasive versus colonizing GBS strains. Methods We screened 162 invasive and 338 colonizing GBS strains from different collections using dot blot hybridization to assess the frequency of bca, bac and rib. All strains were defined by serotyping for capsular type, and frequency differences were tested using the Chi square test. Results Genes encoding the beta C protein (bac and Rib (rib occurred at similar frequencies among invasive and colonizing isolates, bac (20% vs. 23%, and rib (28% vs. 20%, while the alpha (bca C protein was more frequently found in colonizing strains (46% vs, invasive (29%. Invasive strains were associated with specific serotype/gene combinations. Conclusion Novel virulence factors must be identified to better understand GBS disease.

  12. Growth Modeling of Aspergillus niger Strains Isolated from Citrus Fruit as a Function of Temperature on a Synthetic Medium from Lime (Citrus latifolia T.) Pericarp.

    Science.gov (United States)

    Sandoval-Contreras, T; Marín, S; Villarruel-López, A; Gschaedler, A; Garrido-Sánchez, L; Ascencio, F

    2017-07-01

    Molds are responsible for postharvest spoilage of citrus fruits. The objective of this study was to evaluate the effect of temperature on growth rate and the time to visible growth of Aspergillus niger strains isolated from citrus fruits. The growth of these strains was studied on agar lime medium (AL) at different temperatures, and growth rate was estimated using the Baranyi and Roberts model (Int. J. Food Microbiol. 23:277-294, 1994). The Rosso et al. cardinal model with inflexion (L. Rosso, J. R. Lobry, S. Bajard, and J. P. Flandrois, J. Theor. Biol. 162:447-463, 1993) was used as a secondary model to describe the effect of temperature on growth rate and the lag phase. We hypothesized that the same model could be used to calculate the time for the mycelium to become visible (t v ) by substituting the lag phase (1/λ and 1/λ opt ) with the time to visible colony (1/t v -opt and 1/t v ), respectively, in the Rosso et al. High variability was observed at suboptimal conditions. Extremes of temperature of growth for A. niger seem to have a normal variability. For the growth rate and time t v , the model was satisfactorily compared with results of previous studies. An external validation was performed in lime fruits; the bias and accuracy factors were 1.3 and 1.5, respectively, for growth rate and 0.24 and 3.72, respectively, for the appearance time. The discrepancy may be due to the influence of external factors. A. niger grows significantly more slowly on lime fruit than in culture medium, probably because the nutrients are more easily available in medium than in fruits, where the peel consistency may be a physical barrier. These findings will help researchers understand the postharvest behavior of mold on lime fruits, host-pathogen interactions, and environmental conditions infecting fruit and also help them develop guidelines for future work in the field of predictive mycology to improve models for control of postharvest fungi.

  13. New species in Aspergillus section Terrei

    DEFF Research Database (Denmark)

    Samson, R. A.; Peterson, S. W.; Frisvad, Jens Christian

    2011-01-01

    . clade including the type isolate of A. niveus (CBS 115.27) constitutes a lineage closely related to A. carneus. Fennellia nivea, the hypothesized teleomorph is not related to this clade. Aspergillus allahabadii, A. niveus var. indicus, and two species originally placed in section Versicolores, A......Section Terrei of Aspergillus was studied using a polyphasic approach including sequence analysis of parts of the beta-tubulin and calmodulin genes and the ITS region, macro- and micromorphological analyses and examination of extrolite profiles to describe three new species in this section. Based....... floccosus, A. terreus var. africanus, A. terreus var. aureus, while Aspergillus hortai is recognised at species level. Aspergillus terreus NRRL 4017 is described as the new species A. pseudoterreus. Also included in section Terrei are some species formerly placed in sections Flavipedes and Versicolores. A...

  14. Efeito da toxicidade de Cr (VI e Zn (II no crescimento do fungo filamentoso Aspergillus niger isolado de efluente industrial Toxicity effect of Cr (VI and Zn (II on growth of filamentous fungi Aspergillus niger isolated from industrial effluent

    Directory of Open Access Journals (Sweden)

    Maria do Socorro Vale

    2011-09-01

    Full Text Available Processos convencionais de tratamento de efluentes utilizam microrganismos vivos, o que sugere limitações relativas À toxicidade de metais para os microrganismos. O experimento consistiu em adicionar soluções monoelementares de Cr (VI e Zn(II em diferentes concentrações (0, 20, 50, 100, 200, 300, 400, 500 mg.L-1 ao meio de crescimento e observar a influência dos metais no crescimento micelial e germinativo do fungo Aspergillus Níger por verificação visual da expansão radial do micélio e da germinação de esporos, seguida de registro fotográfico. Os resultados mostraram que o metabolismo do fungo foi completamente inibido em concentrações acima de 500 mg Zn (II.L-1 e 150 mg Cr (VI.L-1. O ED50 (concentração de ingrediente ativo capaz de inibir 50% do crescimento micelial do fungo para os dois íons metálicos, nas condições estudadas, está na faixa entre 100 e 150 mg.L-1. Palavras-chave: metais pesados; inibição; crescimento micelial; Aspergillus niger; ED50.Many standard processes of wastewater treatment use live microorganisms, which suggests limitations on a metal toxicity to the microorganism. The experiment consisted in adding mono elementary solutions of Cr (VI and Zn (II at different concentrations (0, 20, 50, 100, 200, 300, 400, 500 mg.L-1 to the growth mean, and to observe the influence of metals on mycelial and germinative growth of the Aspergillus niger fungus, by means of visual observation of the radial expansion of the mycelius and the germination of spores, followed by photograph registration. The results showed that the metabolism of the fungus was completely inhibited at concentrations above 500 mg Zn (II.L-1 and 150 mg Cr (VI.L-1. The ED50 (concentration of active ingredient capable of inhibiting 50% of mycelial growth of the fungus for both metal ions, under the studied conditions, is in the range between 100 and 150 mg.L-1.

  15. Home Environment as a Source of Life-Threatening Azole-Resistant Aspergillus fumigatus in Immunocompromised Patients.

    Science.gov (United States)

    Lavergne, Rose-Anne; Chouaki, Taieb; Hagen, Ferry; Toublanc, Bénédicte; Dupont, Hervé; Jounieaux, Vincent; Meis, Jacques F; Morio, Florent; Le Pape, Patrice

    2017-01-01

    A case of fatal aspergillosis due to a TR 46 /Y121F/T289A azole-resistant Aspergillus fumigatus is reported. Environmental investigations at the patient's residence led to the recovery of TR 46 /Y121F/T289A isolates, genotypically indistinguishable from the clinical isolate, supporting for the first time the direct role of household as potential source of azole-resistant invasive aspergillosis. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

  16. Two novel Aspergillus species from hypersaline soils of The National Park of Lake Urmia, Iran

    DEFF Research Database (Denmark)

    Arzanlou, M.; Samadi, R.; Frisvad, Jens Christian

    2016-01-01

    Two novel Aspergillus species, one belonging to the section Terrei and the other to section Flavipedes, were isolated from hypersaline soils of The National Park of Lake Urmia (Iran) and are here described as Aspergillus iranicus and Aspergillus urmiensis. A polyphasic taxonomic approach comprising...

  17. The role of thiol species in the hypertolerance of Aspergillus sp. P37 to arsenic.

    NARCIS (Netherlands)

    Canovas, D.; Vooijs, H.; Schat, H.; De Lorenzo, V.

    2004-01-01

    Aspergillus sp. P37 is an arsenate-hypertolerant fungus isolated from a river in Spain with a long history of contamination with metals. This strain is able to grow in the presence of 0.2 M arsenate, i.e. 20-fold higher than the reference strain, Aspergillus nidulans TS1. Although Aspergillus sp.

  18. Effect of Environmental Factors on the Growth of Aspergillus Species ...

    African Journals Online (AJOL)

    30 o. C and 35 o. C may be susceptible to contamination by Aspergillus species. ... Aspergillus are common contaminants of food and feed stuffs ... Emerging fungal colonies were continuously sub-cultured on potato dextrose agar plates to obtain pure cultures of the isolates. The fungi were identified based on cultural and ...

  19. Aspergillus Monitoring Project in a Large Educational Hospital ...

    African Journals Online (AJOL)

    Also molecular method, PCR-RFLP using single restriction enzyme as a rapid and available method was performed to investigate environmental sources of Aspergillus infections. Results: Total of 110 clinical fungal isolates included Candida and Aspergillus species and some other opportunistic fungi. Among the clinical

  20. Spectrophotometric reading of EUCAST antifungal susceptibility testing of Aspergillus fumigatus

    NARCIS (Netherlands)

    Meletiadis, J.; Mortensen, K.L.; Verweij, P.E.; Mouton, J.W.; Arendrup, M.C.

    2017-01-01

    OBJECTIVES: Given the increasing number of antifungal drugs and the emergence of resistant Aspergillus isolates, objective, automated and high-throughput antifungal susceptibility testing is important. The EUCAST E.Def 9.3 reference method for MIC determination of Aspergillus species relies on

  1. Exact Molecular Typing of Aspergillus fumigatus. Methods and Applications.

    NARCIS (Netherlands)

    Valk-van Haren, J.A. de

    2008-01-01

    Aspergillus species are widely distributed fungi that release large amounts of airborne conidia that are dispersed in the environment. Aspergillus fumigatus is the species most frequently isolated from human infections. In this thesis a novel assay for fingerprinting A. fumigatus is described and

  2. Global population structure of Aspergillus terreus inferred by ISSR typing reveals geographical subclustering

    Directory of Open Access Journals (Sweden)

    Stevens David A

    2011-09-01

    Full Text Available Abstract Background Aspergillus terreus causes invasive aspergillosis (IA in immunocompromised individuals and can be the leading cause of IA in certain medical centers. We examined a large isolate collection (n = 117 for the presence of cryptic A. terreus species and employed a genome scanning method, Inter-Simple Sequence Repeat (ISSR PCR to determine A. terreus population structure. Results Comparative sequence analyses of the calmodulin locus revealed the presence of the recently recognized species A. alabamensis (n = 4 in this collection. Maximum parsimony, Neighbor joining, and Bayesian clustering of the ISSR data from the 113 sequence-confirmed A. terreus isolates demonstrated that one clade was composed exclusively of isolates from Europe and another clade was enriched for isolates from the US. Conclusions This study provides evidence of a population structure linked to geographical origin in A. terreus.

  3. DNA identification of fetal cells isolated from cervical mucus: potential for early non-invasive prenatal diagnosis.

    Science.gov (United States)

    Katz-Jaffe, Mandy G; Mantzaris, Debbie; Cram, David S

    2005-05-01

    To develop a reliable method to isolate fetal cells for genetic diagnosis. Aspiration of cervical mucus from pregnant women in the first trimester. Pregnant women were recruited before an elective termination of pregnancy. Sixty pregnant women (7-10 weeks of gestation). Fetal cells were isolated from aspirated cervical mucus of pregnant women using a combination of enzymatic digestion, fluorescent immunohistochemistry, micromanipulation and single-cell DNA allelic profiling. The isolation and identification of fetal cells. The transformation of the tenacious cervical mucus into a single-cell suspension enabled the isolation and identification of fetal cells by fluorescent immunohistochemistry. Confirmation of fetal origin was accomplished by single-cell DNA allelic profiling alongside known maternal cells. This novel non-invasive method is rapid and efficient with results attainable within 24 hours as early as seven weeks of gestation. The technique would offer earlier reassurance and the option of first trimester therapeutic abortions to both high and low risk pregnant women.

  4. Conserving a geographically isolated Charaxes butterfly in response to habitat fragmentation and invasive alien plants

    Directory of Open Access Journals (Sweden)

    Casparus J. Crous

    2015-03-01

    Full Text Available In South Africa, much of the forest biome is vulnerable to human-induced disturbance. The forest-dwelling butterfly Charaxes xiphares occidentalis is naturally confined to a small forest region in the south-western Cape, South Africa. Most of the remaining habitat of this species is within a fragmented agricultural matrix. Furthermore, this geographical area is also heavily invaded by alien plants, especially Acacia mearnsii. We investigated how C. x. occidentalis behaviourally responds to different habitat conditions in the landscape. We were particularly interested in touring, patrolling and settling behaviour as a conservation proxy for preference of a certain habitat configuration in this agricultural matrix. Remnant forest patches in the agricultural matrix showed fewer behavioural incidents than in a reference protected area. Moreover, dense stands of A. mearnsii negatively influenced the incidence and settling pattern of this butterfly across the landscape, with fewer tree settlings associated with more heavily invaded forest patches. This settling pattern was predominantly seen in female butterflies. We also identified specific trees that were settled upon for longer periods by C. x. occidentalis. Distance to a neighbouring patch and patch size influenced behavioural incidences, suggesting that further patch degradation and isolation could be detrimental to this butterfly. Conservation implications: We highlight the importance of clearing invasive tree species from vulnerable forest ecosystems and identify key tree species to consider in habitat conservation and rehabilitation programmes for this butterfly. We also suggest retaining as much intact natural forest as possible. This information should be integrated in local biodiversity management plans.

  5. Conserving a geographically isolated Charaxes butterfly in response to habitat fragmentation and invasive alien plants

    Directory of Open Access Journals (Sweden)

    Casparus J. Crous

    2015-08-01

    Full Text Available In South Africa, much of the forest biome is vulnerable to human-induced disturbance. The forest-dwelling butterfly Charaxes xiphares occidentalis is naturally confined to a small forest region in the south-western Cape, South Africa. Most of the remaining habitat of this species is within a fragmented agricultural matrix. Furthermore, this geographical area is also heavily invaded by alien plants, especially Acacia mearnsii. We investigated how C. x. occidentalis behaviourally responds to different habitat conditions in the landscape. We were particularly interested in touring, patrolling and settling behaviour as a conservation proxy for preference of a certain habitat configuration in this agricultural matrix. Remnant forest patches in the agricultural matrix showed fewer behavioural incidents than in a reference protected area. Moreover, dense stands of A. mearnsii negatively influenced the incidence and settling pattern of this butterfly across the landscape, with fewer tree settlings associated with more heavily invaded forest patches. This settling pattern was predominantly seen in female butterflies. We also identified specific trees that were settled upon for longer periods by C. x. occidentalis. Distance to a neighbouring patch and patch size influenced behavioural incidences, suggesting that further patch degradation and isolation could be detrimental to this butterfly. Conservation implications: We highlight the importance of clearing invasive tree species from vulnerable forest ecosystems and identify key tree species to consider in habitat conservation and rehabilitation programmes for this butterfly. We also suggest retaining as much intact natural forest as possible. This information should be integrated in local biodiversity management plans.

  6. Antifungal Susceptibility Testing of Aspergillus spp. by Using a Composite Correlation Index (CCI)-Based Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Method Appears To Not Offer Benefit over Traditional Broth Microdilution Testing.

    Science.gov (United States)

    Gitman, Melissa R; McTaggart, Lisa; Spinato, Joanna; Poopalarajah, Rahgavi; Lister, Erin; Husain, Shahid; Kus, Julianne V

    2017-07-01

    Aspergillus spp. cause serious invasive lung infections, and Aspergillus fumigatus is the most commonly encountered clinically significant species. Voriconazole is considered to be the drug of choice for treating A. fumigatus infections; however, rising resistance rates have been reported. We evaluated a matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based method for the differentiation between wild-type and non-wild-type isolates of 20 Aspergillus spp. (including 2 isolates of Aspergillus ustus and 1 of Aspergillus calidoustus that were used as controls due their intrinsic low azole susceptibility with respect to the in vitro response to voriconazole). At 30 and 48 h of incubation, there was complete agreement between Cyp51A sequence analysis, broth microdilution, and MALDI-TOF MS classification of isolates as wild type or non-wild type. In this proof-of-concept study, we demonstrated that MALDI-TOF MS can be used to accurately detect A. fumigatus strains with reduced voriconazole susceptibility. However, rather than proving to be a rapid and simple method for antifungal susceptibility testing, this particular MS-based method showed no benefit over conventional testing methods. © Crown copyright 2017.

  7. SPECIES OF GENUS ASPERGILLUS ON GRAPE SLOVAK ORIGIN

    Directory of Open Access Journals (Sweden)

    Dana Tančinová

    2014-02-01

    Full Text Available The aim of this study was to detect species of genus Aspergillus from wine grapes (berries, surface sterilized berries - endogenous mycobiota, from damaged berries and grape juice of Slovak origin. We analyzed 20 samples of grapes, harvested in 2011 from various wine-growing regions. For the isolation of species we used the method of direct plating berries, surface-sterilized berries (using 0.4% freshly pre-pared chlorine, and damaged berries on DRBC (Dichloran Rose Bengal Chloramphenicol agar. For the determination of fungal contamination of grape juice, we used plate-dilution method and DRBC as medium. The cultivation in all modes of inoculation was carried at 25±1°C, for 5 to 7 days. After incubation Aspergillus isolates were inoculated on the identification media. Representatives of the genus Aspergillus were isolated from 13 samples berries, 7 samples of surface-sterilized berries, 4 samples of damaged berries and 9 samples of grape juice. Overall, representatives of aspergilli were detected in 90% of samples (75 isolates. In this work we focused on the detection of potential producers of ochratoxin A belonging to the genus Aspergillus. Isolates, potential producers of ochratoxin A (Aspergillus niger aggregate and Aspergillus westerdijkiae, were after their identification inoculated on YES medium (Yeast Extract Sucrose Agar and after 14 days of incubation at 25±1°C, in the dark, we tested them for their ability to produce ochratoxin A using thin layer chromatography. Out of the 16 isolates from isolated potential producers of ochratoxin A none of the isolates of Aspergillus niger aggregate (13 tested produced ochratoxin A. The isolate of Aspergillus westerdijkiae (1, isolated from the surface-sterilized berries, produced ochratoxin A.

  8. Transplant tourism and invasive fungal infection.

    Science.gov (United States)

    Al Salmi, I; Metry, A M; Al Ismaili, F; Hola, A; Al Riyami, M; Khamis, F; Al-Abri, S

    2018-04-01

    Deceased and live-related renal transplants (RTXs) are approved procedures that are performed widely throughout the world. In certain regions, commercial RTX has become popular, driven by financial greed. This retrospective, descriptive study was performed at the Royal Hospital from 2013 to 2015. Data were collected from the national kidney transplant registry of Oman. All transplant cases retrieved were divided into two groups: live-related RTX performed in Oman and commercial-unrelated RTX performed abroad. These groups were then divided again into those with and without evidence of fungal infection, either in the wound or renal graft. A total of 198 RTX patients were identified, of whom 162 (81.8%) had undergone a commercial RTX that was done abroad. Invasive fungal infections (IFIs) were diagnosed in 8% of patients who had undergone a commercial RTX; of these patients, 76.9% underwent a nephrectomy and 23.1% continued with a functioning graft. None of the patients with RTXs performed at the Royal Hospital contracted an IFI. The most common fungal isolates were Aspergillus species (including Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, and Aspergillus nigricans), followed by Zygomycetes. However, there was no evidence of fungal infection including Aspergillus outside the graft site. Computed tomography (CT) findings showed infarction of the graft, renal artery thrombosis, aneurysmal dilatation of the external iliac artery, fungal ball, or just the presence of a perigraft collection. Of the total patients with IFIs, 23.1% died due to septic shock and 53.8% were alive and on hemodialysis. The remaining 23.1% who did not undergo nephrectomy demonstrated acceptable graft function. This is the largest single-center study on commercial RTX reporting the highest number of patients with IFI acquired over a relatively short period of time. Aspergillus spp were the main culprit fungi, with no Candida spp being isolated. A high index of suspicion might

  9. Antifungal activity of terrestrial Streptomyces rochei strain HF391 against clinical azole -resistant Aspergillus fumigatus

    Science.gov (United States)

    Hadizadeh, S; Forootanfar, H; Shahidi Bonjar, GH; Falahati Nejad, M; Karamy Robati, A; Ayatollahi Mousavi, SA; Amirporrostami, S

    2015-01-01

    Background and Purpose: Actinomycetes have been discovered as source of antifungal compounds that are currently in clinical use. Invasive aspergillosis (IA) due to Aspergillus fumigatus has been identified as individual drug-resistant Aspergillus spp. to be an emerging pathogen opportunities a global scale. This paper described the antifungal activity of one terrestrial actinomycete against the clinically isolated azole-resistant A. fumigatus. Materials and Methods: Soil samples were collected from various locations of Kerman, Iran. Thereafter, the actinomycetes were isolated using starch-casein-nitrate-agar medium and the most efficient actinomycetes (capable of inhibiting A. fumigatus) were screened using agar block method. In the next step, the selected actinomycete was cultivated in starch-casein- broth medium and the inhibitory activity of the obtained culture broth was evaluated using agar well diffusion method. Results: The selected actinomycete, identified as Streptomyces rochei strain HF391, could suppress the growth of A. fumigatus isolates which was isolated from the clinical samples of patients treated with azoles. This strain showed higher inhibition zones on agar diffusion assay which was more than 15 mm. Conclusion: The obtained results of the present study introduced Streptomyces rochei strain HF391 as terrestrial actinomycete that can inhibit the growth of clinically isolated A. fumigatus. PMID:28680984

  10. Isolation and characterization of a native strain of Aspergillus niger ZRS14 with capability of high resistance to zinc and its supernatant application towards extracellular synthesis of zinc oxide nanoparticles

    Directory of Open Access Journals (Sweden)

    Morahem Ashengroph

    2013-01-01

    Full Text Available Introduction: Zinc oxide nanoparticles have quite a few applications in the fields of biology, optics, mechanics, magnetism, energy, hygiene and medicine. Due to serious problems associated with physiochemical synthesis of ZnO nanoparticles, including environmental pollution, complicated and costly processes, there is a growing need to develop a simple biological procedure for synthesis of nanoparticles to achieve the monodisperse-sized particles with a higher purity, low energy consumption and a cleaner environment. We conducted this investigation to screen and isolate native fungi strains capable of high zinc metal tolerance ability and a potential for extracellular synthesis of ZnO nanoparticles using fungal secretions as biological catalysts.Materials and methods: 15 different strains of fungi were isolated from soil samples collected from lead and zinc mines of Angoran-Zanjan using conventional enrichment process and characterized initially based on macroscopic and microscopic characteristics and colony morphology. The intrinsic tolerance of the isolated strains to zinc toxic metal was measured in the synthetic and complex media using the agar dilution method. The supernatants of isolated fungi were incubated with zinc acetate solution in a shaker incubator for 72h; then, the strain that was able to synthesis ZnO nanoparticle was identified. The ZnO nanoparticles formation was investigated by using spectroscopic techniques and microscopic observations.Results: Among the 15 isolated strains, the strain ZRS14 had highest zinc metal tolerance ability and was selected and identified as Aspergillus niger strain ZRS14 (GenBank accession number KF414527 based on morphological and molecular phylogenetic analysis. For synthesis of ZnO nanoparticles by isolated A. niger ZRS14, fungal cell-free filtrate of the strain was collected and incubated in the presence of zinc acetate solution at a final concentration of 250 mg/l zinc metal ion at 28º C for

  11. Prevalence and mechanism of triazole resistance in Aspergillus fumigatus in a referral chest hospital in Delhi, India and an update of the situation in Asia

    Directory of Open Access Journals (Sweden)

    Anuradha eChowdhary

    2015-05-01

    Full Text Available Aspergillus fumigatus causes varied clinical syndromes ranging from colonization to deep infections. The mainstay of therapy of Aspergillus diseases is triazoles but several studies globally highlighted variable prevalence of triazole resistance, which hampers the management of aspergillosis. We studied the prevalence of resistance in clinical A. fumigatus isolates during 4 years in a referral Chest Hospital in Delhi, India and reviewed the scenario in Asia and the Middle East. Aspergillus species (n=2117 were screened with selective plates for azole resistance. The isolates included 45.4% A. flavus, followed by 32.4% A. fumigatus, 15.6% Aspergillus species and 6.6% A. terreus. Azole resistance was found in only 12 (1.7% A. fumigatus isolates.These triazole resistant A. fumigatus (TRAF isolates were subjected to (a calmodulin and β tubulin gene sequencing (b in vitro antifungal susceptibility testing against triazoles using CLSI M38-A2 (c sequencing of cyp51A gene and real-time PCR assay for detection of mutations and (d microsatellite typing of the resistant isolates. TRAF harbored TR34/L98H mutation in 10 (83.3% isolates with a pan-azole resistant phenotype. Among the remaining 2 TRAF isolates, one had G54E and the other had three non-synonymous point mutations. The majority of patients were diagnosed as invasive aspergillosis followed by allergic bronchopulmonary aspergillosis and chronic pulmonary aspergillosis. The Indian TR34/L98H isolates had a unique genotype and were distinct from the Chinese, Middle East and European TR34/L98H strains. This resistance mechanism has been linked to the use of fungicide azoles in agricultural practices in Europe as it has been mainly reported from azole naïve patients. Reports published from Asia demonstrate the same environmental resistance mechanism in A. fumigatus isolates from two highly populated countries in Asia i.e., China and India and also from the neighboring Middle East.

  12. Isavuconazole versus voriconazole for primary treatment of invasive mould disease caused by Aspergillus and other filamentous fungi (SECURE): a phase 3, randomised-controlled, non-inferiority trial.

    Science.gov (United States)

    Maertens, Johan A; Raad, Issam I; Marr, Kieren A; Patterson, Thomas F; Kontoyiannis, Dimitrios P; Cornely, Oliver A; Bow, Eric J; Rahav, Galia; Neofytos, Dionysios; Aoun, Mickael; Baddley, John W; Giladi, Michael; Heinz, Werner J; Herbrecht, Raoul; Hope, William; Karthaus, Meinolf; Lee, Dong-Gun; Lortholary, Olivier; Morrison, Vicki A; Oren, Ilana; Selleslag, Dominik; Shoham, Shmuel; Thompson, George R; Lee, Misun; Maher, Rochelle M; Schmitt-Hoffmann, Anne-Hortense; Zeiher, Bernhardt; Ullmann, Andrew J

    2016-02-20

    Isavuconazole is a novel triazole with broad-spectrum antifungal activity. The SECURE trial assessed efficacy and safety of isavuconazole versus voriconazole in patients with invasive mould disease. This was a phase 3, double-blind, global multicentre, comparative-group study. Patients with suspected invasive mould disease were randomised in a 1:1 ratio using an interactive voice-web response system, stratified by geographical region, allogeneic haemopoietic stem cell transplantation, and active malignant disease at baseline, to receive isavuconazonium sulfate 372 mg (prodrug; equivalent to 200 mg isavuconazole; intravenously three times a day on days 1 and 2, then either intravenously or orally once daily) or voriconazole (6 mg/kg intravenously twice daily on day 1, 4 mg/kg intravenously twice daily on day 2, then intravenously 4 mg/kg twice daily or orally 200 mg twice daily from day 3 onwards). We tested non-inferiority of the primary efficacy endpoint of all-cause mortality from first dose of study drug to day 42 in patients who received at least one dose of the study drug (intention-to-treat [ITT] population) using a 10% non-inferiority margin. Safety was assessed in patients who received the first dose of study drug. This study is registered with ClinicalTrials.gov, number NCT00412893. 527 adult patients were randomly assigned (258 received study medication per group) between March 7, 2007, and March 28, 2013. All-cause mortality from first dose of study drug to day 42 for the ITT population was 19% with isavuconazole (48 patients) and 20% with voriconazole (52 patients), with an adjusted treatment difference of -1·0% (95% CI -7·8 to 5·7). Because the upper bound of the 95% CI (5·7%) did not exceed 10%, non-inferiority was shown. Most patients (247 [96%] receiving isavuconazole and 255 [98%] receiving voriconazole) had treatment-emergent adverse events (p=0·122); the most common were gastrointestinal disorders (174 [68%] vs 180 [69%]) and infections and

  13. Aspergillus baeticus sp. nov. and Aspergillus thesauricus sp. nov., two species in section Usti from Spanish caves.

    Science.gov (United States)

    Nováková, Alena; Hubka, Vit; Saiz-Jimenez, Cesareo; Kolarik, Miroslav

    2012-11-01

    Two novel species of Aspergillus that are clearly distinct from all known species in section Usti were revealed during a study of microfungal communities in Spanish caves. The novel species identified in this study and additional species of Aspergillus section Usti are associated with places and substrates related to human activities in caves. Novel species are described using data from four loci (ITS, benA, caM and rpb2), morphology and basic chemical and physiological analyses. Members of the species Aspergillus thesauricus sp. nov. were isolated from various substrates, including decaying organic matter, cave air and cave sediment of the Cueva del Tesoro Cave (the Treasure cave); the species is represented by twelve isolates and is most closely related to the recently described Aspergillus germanicus. Members of the species Aspergillus baeticus sp. nov. were isolated from cave sediment in the Gruta de las Maravillas Cave (the Grotto of the Marvels); the species is represented by two isolates. An additional isolate was found in the Cueva del Tesoro Cave and in the Demänovská Peace Cave (Slovakia), suggesting a potentially wide distribution of this micro-organism. The species is related to Aspergillus ustus and Aspergillus pseudoustus. Both species were unable to grow at 37 °C, and a weakly positive, light greenish yellow Ehrlich reaction was observed in A. thesauricus. Unique morphological features alone are sufficient to distinguish both species from related taxa.

  14. Pseudomembranous aspergillus bronchitis in a double-lung transplanted patient: unusual radiographic and CT features

    International Nuclear Information System (INIS)

    Ducreux, D.; Chevallier, P.; Raffaelli, C.; Padovani, B.; Perrin, C.; Jourdan, J.; Hofman, P.

    2000-01-01

    Pseudomembranous aspergillus bronchitis is considered as an early form of invasive pulmonary aspergillosis, a well-known airway infection in immunocompromised patients. Radiologic features concerning invasive aspergillosis of the airways have been reported. However, we describe here an unusual feature of invasive aspergillus bronchitis, never reported to date, observed in a double-lung transplanted patient. Chest radiograph and CT revealed significant peribronchial thickening without any parenchymal involvement. (orig.)

  15. Pectinase and polygalacturonase production by a thermophilic Aspergillus fumigatus isolated from decomposting orange peels Produção de pectinases e poligalacturonase por Aspergillus fumigatus termofílico isolado de cascas de laranja em decomposição

    Directory of Open Access Journals (Sweden)

    Urmila Phutela

    2005-03-01

    Full Text Available A thermophilic fungal strain producing both pectinase and polygalacturonase was isolated after primary screening of 120 different isolates. The fungus was identified as Aspergillus fumigatus Fres. MTCC 4163. Using solid-state cultivation, the optimum levels of variables for pectinase and polygalacturonase (PG production were determined. Maximal levels of enzyme activities were achieved upon growing the culture in a medium containing wheat bran, sucrose, yeast extract and (NH42SO4 after 2-3 days of incubation at a temperature of 50ºC. Highest enzyme activities of 1116 Ug-1 for pectinase and 1270 Ug-1 for polygalacturonase were obtained at pH 4.0 and 5.0, respectively.Através da tiragem de 120 cepas de fungos, isolou-se uma cepa capaz de produzir tanto pectinase quanto poligalacturonase. A cepa foi identificada como Aspergillus fumigatus Fres. MTCC 4163. Empregando cultivo em estado sólido, determinou-se os níveis ótimos das variáveis para a produção de pectinase e de poligalacturonase. Os níveis máximos de atividade enzimática foram obtidos quando a cultura era realizada em meio contendo farelo de trigo, sacarose, extrato de levedura e (NH42SO4 por 2-3 dias a uma temperatura de 50ºC. A atividade máxima de pectinase (1116 Ug-1 e de poligalacturonase (1270 Ug-1 foi obtida em pH 4,0 e 5,0, respectivamente.

  16. Optimization of chloroxylenol degradation by Aspergillus niger using ...

    African Journals Online (AJOL)

    Burman design and response surface methodology. ... Abstract. Chloroxylenol is a very toxic phenolic derivative and it represents potential hazard towards human health and to the environment. Aspergillus niger, local isolate, is an efficient fungus ...

  17. Uptake and efflux kinetics, and intracellular activity of voriconazole against Aspergillus fumigatus in human pulmonary epithelial cells: a new application for the prophylaxis and early treatment of invasive pulmonary aspergillosis.

    Science.gov (United States)

    Wang, Taotao; Yang, Qianting; Chen, Lu; Li, Ying; Meng, Ti; Wang, Yan; Zhang, Tao; Lei, Jin'e; Xing, Jianfeng; Dong, Yalin

    2017-06-01

    Invasive pulmonary aspergillosis (IPA), most caused by Aspergillus fumigatus, is a serious life-threatening infection in immunocompromised patients. Voriconazole is used to prevent and treat IPA. However, little is known about the pharmacological characteristics of voriconazole in pulmonary epithelial cells, which are the target site for the prophylaxis and early treatment of IPA. The aim of the study was to evaluate the kinetics and activity of voriconazole against A. fumigatus in A549 cells. High-performance liquid chromatography/tandem mass spectrometry and time-kill method were used to study the cellular pharmacokinetic and pharmacodynamics of voriconazole. Voriconazole exerted a concentration-dependent toxic effect on A549 cells and could penetrate into cells, reaching plateau concentrations of 1.14 ± 0.64, 3.72 ± 1.38 and 6.36 ± 0.95 ng/mg protein after A549 cells were exposed to voriconazole at extracellular concentrations of 2, 8 and 16 mg/L for 2 h, respectively. The efflux of voriconazole was rapid, with a half-life of 10.2 min. Voriconazole can decrease the A. fumigatus conidia invade cells, and the number of viable A. fumigatus conidia in cells can be decreased 2.1- to 20.6-fold when A549 cells were cultured in medium containing voriconazole. After 24-h incubation, 75.6% and 80.5% of intracellular A. fumigatus were killed when extracellular voriconazole concentration was 8 and 16 mg/L, respectively. This study illustrated a new application for the prophylaxis and early treatment of IPA from the cellular pharmacokinetics and pharmacodynamics and emphasized the importance of monitoring concentrations of voriconazole in epithelial lining fluid in immunocompromised patients receiving voriconazole therapy. © 2016 Société Française de Pharmacologie et de Thérapeutique.

  18. Trends in Antimicrobial Resistance in 1,968 Invasive Streptococcus pneumoniae Strains Isolated in Spanish Hospitals (2001 to 2003): Decreasing Penicillin Resistance in Children's Isolates

    Science.gov (United States)

    Oteo, Jesús; Lázaro, Edurne; de Abajo, Francisco J.; Baquero, Fernando; Campos, José

    2004-01-01

    To address the public health problem of antibiotic resistance, the European Union (EU) founded the European Antimicrobial Resistance Surveillance System. A network of 40 hospitals that serve approximately 30% of the Spanish population (about 12 million) participated. Each laboratory reported data on antimicrobial susceptibility testing using standard laboratory procedures that were evaluated by an external quality control program. The antibiotic consumption data were obtained from the National Health System. We compared the antibiotic susceptibility of Spanish isolates of invasive Streptococcus pneumoniae (2001 to 2003) with antibiotic consumption. Invasive S. pneumoniae was isolated from 1,968 patients, 20% of whom were children at or below the age of 14 years. Of non-penicillin-susceptible strains (35.6%; 95% confidence interval, 34 to 37.2), 26.4% were considered intermediate and 9.2% were considered resistant. Between 2001 and 2003, penicillin resistance decreased from 39.5 to 33% overall and from 60.4 to 41.2% in children at or below the age of 14 years (P = 0.002). Resistance to erythromycin was at 26.6%, and coresistance with penicillin was at 19.1%. Of total isolates, the ciprofloxacin MIC was >2 μg/ml for 2.1%, with numbers increasing from 0.4% (2001) to 3.9% (2003). Total antibiotic use decreased from 21.66 to 19.71 defined daily doses/1,000 inhabitants/day between 1998 and 2002. While consumption of broad-spectrum penicillins, cephalosporins, and erythromycin declined, use of amoxicillin-clavulanate and quinolones increased by 17.5 and 27%, respectively. The frequency of antibiotic resistance in invasive S. pneumoniae in Spain was among the highest in the EU. However, a significant decrease in penicillin resistance was observed in children. This decrease coincided with the introduction of a heptavalent conjugate pneumoccocal vaccine (June 2001) and with a global reduction in antibiotic consumption levels. PMID:15583283

  19. Polyphasic taxonomy of Aspergillus section Cervini

    DEFF Research Database (Denmark)

    Chen, A.J.; Varga, J.; Frisvad, Jens Christian

    2016-01-01

    Species belonging to Aspergillus section Cervini are characterised by radiate or short columnar, fawn coloured, uniseriate conidial heads. The morphology of the taxa in this section is very similar and isolates assigned to these species are frequently misidentified. In this study, a polyphasic ap...

  20. Immuno-polymerase chain reaction for detection ofAspergillus fumigatus

    OpenAIRE

    Sarma, P. Usha; Bir, Nivedita; Paliwal, Anubha; Reddy, Prasad

    1997-01-01

    A number of Aspergillus infections are caused by the opportunistic fungal pathogenAspergillus fumigatus in humans especially under immunosuppressed conditions. Major forms of the disease include invasive aspergillosis, allergic bronchopulmonary aspergillosis and aspergilloma. A procedure that uses chitinase and microwave treatment is described for the extraction of genomic DNA of Aspergillus species from the sputum and bronchial aspirate of patients with established aspergillosis. Detection o...

  1. QM/MM investigation of the reaction rates of substrates of 2,3-dimethylmalate lyase: A catabolic protein isolated from Aspergillus niger.

    Science.gov (United States)

    Chotpatiwetchkul, Warot; Jongkon, Nathjanan; Hannongbua, Supa; Gleeson, M Paul

    2016-07-01

    Aspergillus niger is an industrially important microorganism used in the production of citric acid. It is a common cause of food spoilage and represents a health issue for patients with compromised immune systems. Recent studies on Aspergillus niger have revealed details on the isocitrate lyase (ICL) superfamily and its role in catabolism, including (2R, 3S)-dimethylmalate lyase (DMML). Members of this and related lyase super families are of considerable interest as potential treatments for bacterial and fungal infections, including Tuberculosis. In our efforts to better understand this class of protein, we investigate the catalytic mechanism of DMML, studying five different substrates and two different active site metals configurations using molecular dynamics (MD) and hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. We show that the predicted barriers to reaction for the substrates show good agreement with the experimental kcat values. This results help to confirm the validity of the proposed mechanism and open up the possibility of developing novel mechanism based inhibitors specifically for this target. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. A New Pair of Pentaketide Diastereoisomers from Aspergillus melleus YIM PHI001

    Directory of Open Access Journals (Sweden)

    Qiang Wang

    2018-05-01

    Full Text Available Two new aspinotriol derivatives (1-2 determined as melleusin A (1, B (2 and other seven known compounds were isolated from a soil-borne fungus Aspergillus melleus. The configurations of 1, and 2 were determined by their analogues, aspilactonol B, C isolated previous in Aspergillus. Mellein (8 showed broad antibiotic activity against the test pathogens. Aspinonene, aspinotriols A and B can be used as the marker molecule in chemotaxonomy of Aspergillus.

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

    Science.gov (United States)

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

    2014-10-01

    In a previous study we have shown that the in vitro invasion rate (IR) and tachyzoite yield (TY) are associated with the virulence phenotypes of Neospora caninum isolates of bovine origin. In addition, we recently observed marked differences in virulence when canine isolates were compared in a pregnant BALB/c mouse model. In this study, we investigated whether invasion and proliferation capacities could be used as virulence-related N. caninum phenotypic traits. Of the isolates compared in mice, four canine isolates obtained from oocysts (Nc-Ger2, Nc-Ger3, Nc-Ger-6, Nc-6 Arg) had shown a low-moderate virulence, and two further isolates obtained from dogs with neurological signs (Nc-Bahia, Nc-Liv) were highly virulent. The IR for each isolate was determined by a plaque assay and the counting of immunofluorescence-labeled parasitophorous vacuoles at 3 days post-inoculation (p.i.). The TY was determined by the quantification of tachyzoites at 56 h p.i. by real-time PCR. Most of the canine isolates showed similar IR values under controlled invasion conditions for 4h and 72 h p.i., indicating a limited time period for invasion similar to that observed for bovine isolates. The Nc-Ger3, Nc-Bahia, and Nc-Liv isolates showed a significantly higher IR and TY than the Nc-Ger2 and Nc-Ger6 isolates (P0.885, P0.8117, P<0.058) in mice. These results demonstrate the importance both the invasive and proliferative capacities have on the virulence of canine N. caninum isolates. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. Isolation and characterization of polymorphic microsatellite loci from the invasive plant Solidago canadensis (Asteraceae).

    Science.gov (United States)

    Zhao, S-Y; Sun, S-G; Guo, Y-H; Chen, J-M; Wang, Q-F

    2012-02-17

    Solidago canadensis, a clonal herb originally from North America (common name: Canada goldenrod), is an invasive species in many countries. We developed microsatellite primers for this species. Eleven polymorphic loci were generated and primers were designed. Polymorphism of these 11 loci was assessed in 35 plants from two populations (Wuhan and Shanghai) in China. The number of alleles per locus ranged from 3 to 14. The observed and expected heterozygosities varied from 0.0732 to 0.7391 and from 0.1177 to 0.8687, respectively. These microsatellite markers will be useful tools for studies of population genetics in the native and invasive range of this species.

  5. Progressive Increase in Antimicrobial Resistance among Invasive Isolates of Haemophilus influenzae Obtained from Children Admitted to a Hospital in Kilifi, Kenya, from 1994 to 2002

    Science.gov (United States)

    Scott, J. Anthony G.; Mwarumba, Salim; Ngetsa, Caroline; Njenga, Salome; Lowe, Brett S.; Slack, Mary P. E.; Berkley, James A.; Mwangi, Isaiah; Maitland, Kathryn; English, Mike; Marsh, Kevin

    2005-01-01

    Etest susceptibilities to amoxicillin, chloramphenicol, and trimethoprim-sulfamethoxazole of 240 invasive isolates of Haemophilus influenzae cultured from children in rural Kenya were 66%, 66%, and 38%, respectively. Resistance increased markedly over 9 years and was concentrated among serotype b isolates. In Africa, the increasing cost of treating resistant infections supports economic arguments for prevention through conjugate H. influenzae type b immunization. PMID:15980390

  6. Sentinel Lymph Node Biopsy and Isolated Tumor Cells in Invasive Lobular Versus Ductal Breast Cancer

    NARCIS (Netherlands)

    Truin, Wilfred; Roumen, Rudi M.; Siesling, Sabine; van der Heiden-van der Loo, Margriet; Lobbezoo, Dorien J.; Tjan-Heijnen, Vivianne C.G.; Voogd, Adri C.

    2016-01-01

    Background Sentinel lymph node (SLN) biopsy is the standard of care for axillary staging in invasive breast cancer. The introduction of SLN biopsy with an extensive pathology examination, in addition to the introduction of the 2002 TNM classification, led to different axillary classification

  7. Invasion versus isolation: Trade-offs in managing native salmonids with barriers to upstream movement

    Science.gov (United States)

    Kurt D. Fausch; Bruce E. Rieman; Jason B. Dunham; Michael K. Young; Douglas P. Peterson

    2009-01-01

    Conservation biologists often face the trade-off that increasing connectivity in fragmented landscapes to reduce extinction risk of native species can foster invasion by non-native species that enter via the corridors created, which can then increase extinction risk. This dilemma is acute for stream fishes, especially native salmonids, because their populations are...

  8. Clonal relationship of recent invasive Haemophilus influenzae serotype f isolates from Denmark and the United States

    DEFF Research Database (Denmark)

    Bruun, B; Gahrn-Hansen, B; Westh, H

    2004-01-01

    Surveillance performed after the introduction of general Haemophilus influenzae serotype b (Hib) vaccination in Denmark identified 13 cases of invasive bacteraemic H. influenzae serotype f (Hif) disease in adults over a period of 7 years. Bacteraemic respiratory tract infections accounted for 61...

  9. Kefir-isolated bacteria and yeasts inhibit Shigella flexneri invasion and modulate pro-inflammatory response on intestinal epithelial cells.

    Science.gov (United States)

    Bolla, P A; Abraham, A G; Pérez, P F; de Los Angeles Serradell, M

    2016-02-01

    The aim of this work was to evaluate the ability of a kefir-isolated microbial mixture containing three bacterial and two yeast strains (MM) to protect intestinal epithelial cells against Shigella flexneri invasion, as well as to analyse the effect on pro-inflammatory response elicited by this pathogen. A significant decrease in S. flexneri strain 72 invasion was observed on both HT-29 and Caco-2 cells pre-incubated with MM. Pre-incubation with the individual strains Saccharomyces cerevisiae CIDCA 8112 or Lactococcus lactis subsp. lactis CIDCA 8221 also reduced the internalisation of S. flexneri into HT-29 cells although in a lesser extent than MM. Interestingly, Lactobacillus plantarum CIDCA 83114 exerted a protective effect on the invasion of Caco-2 and HT-29 cells by S. flexneri. Regarding the pro-inflammatory response on HT-29 cells, S. flexneri infection induced a significant activation of the expression of interleukin 8 (IL-8), chemokine (C-C motif) ligand 20 (CCL20) and tumour necrosis factor alpha (TNF-α) encoding genes (Pkefir, resulted in inhibition of S. flexneri internalisation into human intestinal epithelial cells, along with the inhibition of the signalling via NF-κB that in turn led to the attenuation of the inflammatory response.

  10. Host age and expression of genes involved in red blood cell invasion in Plasmodium falciparum field isolates.

    Science.gov (United States)

    Valmaseda, Aida; Bassat, Quique; Aide, Pedro; Cisteró, Pau; Jiménez, Alfons; Casellas, Aina; Machevo, Sonia; Aguilar, Ruth; Sigaúque, Betuel; Chauhan, Virander S; Langer, Christine; Beeson, James; Chitnis, Chetan; Alonso, Pedro L; Gaur, Deepak; Mayor, Alfredo

    2017-07-05

    Plasmodium falciparum proteins involved in erythrocyte invasion are main targets of acquired immunity and important vaccine candidates. We hypothesized that anti-parasite immunity acquired upon exposure would limit invasion-related gene (IRG) expression and affect the clinical impact of the infection. 11 IRG transcript levels were measured in P. falciparum isolates by RT-PCR, and IgG/IgM against invasion ligands by Luminex®, in 50 Mozambican adults, 25 children with severe malaria (SM) and 25 with uncomplicated malaria (UM). IRG expression differences among groups and associations between IRG expression and clinical/immunologic parameters were assessed. IRG expression diversity was higher in parasites infecting children than adults (p = 0.022). eba140 and ptramp expression decreased with age (p = 0.003 and 0.007, respectively) whereas p41 expression increased (p = 0.022). pfrh5 reduction in expression was abrupt early in life. Parasite density decreased with increasing pfrh5 expression (p immune escape when tested in malaria-exposed individuals.

  11. Pyrophen Produced by Endophytic Fungi Aspergillus sp Isolated from Piper crocatum Ruiz and Pav Exhibits Cytotoxic Activity and Induces S Phase Arrest in T47D Breast Cancer Cells.

    Science.gov (United States)

    Astuti, Puji; Erden, Willy; Wahyono; Wahyuono, Subagus; Hertiani, Triana

    2016-01-01

    Ethyl acetate extracts obtained from culture of endophytic fungi Aspergillus sp isolated from Piper crocatum Ruiz and Pav, have been shown to possess cytotoxic activity against T47D breast cancer cells. Investigations were here conducted to determine bioactive compounds responsible for the activity. Bioassay guided fractionation was employed to obtain active compounds. Structure elucidation was performed based on analysis of LC-MS, 1H-NMR, 13C-NMR, COSY, DEPT, HMQC, HMBC data. Cytotoxity assays were conducted in 96 well plates against T47D and Vero cell lines. Bioassay guided isolation and chemical investigation led to the isolation of pyrophen, a 4-methoxy-6-(1'-acetamido-2'-phenylethyl)-2H-pyran-2-one. Further analysis of its activity against T47D and Vero cells showed an ability to inhibit the growth of T47D cells with IC50 values of 9.2 μg/mL but less cytotoxicity to Vero cells with an IC50 of 109 μg/mL. This compound at a concentration of 400 ng/mL induced S-phase arrest in T47D cells.

  12. Changing trends in serotypes of S. pneumoniae isolates causing invasive and non-invasive diseases in unvaccinated population in Mexico (2000-2014).

    Science.gov (United States)

    Carnalla-Barajas, María Noemí; Soto-Noguerón, Araceli; Sánchez-Alemán, Miguel Angel; Solórzano-Santos, Fortino; Velazquez-Meza, María Elena; Echániz-Aviles, Gabriela

    2017-05-01

    Introduction of pneumococcal conjugate vaccines (PCV) targeted against a limited number of serotypes substantially decreased invasive (IPD) and non-invasive pneumococcal diseases (NIPD) but it was accompanied by non-vaccine type replacement disease. After 9 years of introduction of PCV in Mexico, we analyze the evidence of the indirect effects on IPD and NIPD serotype distribution among groups not targeted to receive the vaccine. From January 2000 to December 2014, pneumococcal strains isolated from IPD and NIPD cases from patients ≥5 years of age from participant hospitals of the SIREVA II (Sistema Regional de Vacunas) network were serotyped. A regression analysis was performed considering year and proportion of serotypes included in the different vaccine formulations (PCV7, PCV10 and PCV13). The slope was obtained for each regression line and their correspondent p-value. The proportion of each serotype in the pre-PCV7 and post-PCV7 periods was evaluated by χ2 test. From a total of 1147 pneumococcal strains recovered, 570 corresponded to the pre-PCV7 and 577 to the post-PCV7 periods. The proportion of vaccine serotypes included in the three PCV formulations decreased by 2.4, 2.6 and 1.3%, respectively per year during the study period. A significant increase of serotype 19A was observed in the post-vaccine period in all age groups. A percentage of annual decline of serotypes causing IPD and NIPD included in PCV was detected among groups not targeted to receive the vaccine, probably due to herd effect. Considering pneumococcal serotype distribution is a dynamic process, we highlight the importance of surveillance programs. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  13. Biosorption characteristics of Aspergillus fumigatus in removal of ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-09-01

    Sep 1, 2009 ... Nineteen fungal species were isolated from soil contaminated with industrial wastes of which. Aspergillus species were the most dominant. The growth of the isolates was notice by Cd concentration in growth medium, thus about 20% of the isolates can grow up to 50 mg Cd/100 ml medium and only.

  14. Aspergillus spondylodiscitis in a patient with liver cirrhosis and diabetes mellitus

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    Tereza Rojko

    2014-11-01

    Full Text Available Aspergillus osteomyelitis is a rare manifestation of invasive aspergillosis which accounts for 1.8% to 5.6% of all invasive aspergillosis forms. As other forms of invasive aspergillosis, it predominantly occurs in immunosupressed patients with well established risk factors, but there are increasing reports of invasive aspergillosis cases in patients without traditional risk factors such as patients with chronic obstructive pulmonary disease on inhalatory corticosteroid therapy or patients with liver cirrhosis. We present a case of Aspergillus spondylodiscitis in a patient without other risk factors for invasive aspergillosis than liver cirrhosis and diabetes mellitus successfully treated with a 4-month course of voriconasole.

  15. A study on accumulation of volatile organic compounds during ochratoxin a biosynthesis and characterization of the correlation in Aspergillus carbonarius isolated from grape and dried vine fruit.

    Science.gov (United States)

    Zhang, Xiaoxu; Cheng, Zhan; Ma, Liyan; Li, Jingming

    2017-07-15

    Volatile organic compounds (VOCs) and ochratoxin A (OTA) were detected in Aspergillus carbonarius strains (AF, SD26 and SD27) during a 7-day growing period, using HS-SPME-GC-MS and HPLC-MS/MS, respectively. Results showed that 33 VOCs were detected in these strains. The AF strain showed the different sesquiterpene profile compared with the other strains. Principal component analysis revealed that the AF strain was segregated from the other two strains regarding the VOC profile, especially the sesquiterpene profile. Ester metabolites were produced more at the beginning of the strain growth period, whereas extension of the strain incubation resulted in an intense accumulation of hydrocarbon compounds. The prediction models on the correlation between OTA and VOCs were established using partial least squares regression analysis. This analysis indicated that C-8 alcohols, ketones, and trans-nerolidol showed a close relation with the OTA synthesis. Copyright © 2016. Published by Elsevier Ltd.

  16. Oral Candida albicans isolates from HIV-positive individuals have similar in vitro biofilm-forming ability and pathogenicity as invasive Candida isolates

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    Rasteiro Vanessa MC

    2011-11-01

    Full Text Available Abstract Background Candida can cause mucocutaneous and/or systemic infections in hospitalized and immunosuppressed patients. Most individuals are colonized by Candida spp. as part of the oral flora and the intestinal tract. We compared oral and systemic isolates for the capacity to form biofilm in an in vitro biofilm model and pathogenicity in the Galleria mellonella infection model. The oral Candida strains were isolated from the HIV patients and included species of C. albicans, C. glabrata, C. tropicalis, C. parapsilosis, C. krusei, C. norvegensis, and C. dubliniensis. The systemic strains were isolated from patients with invasive candidiasis and included species of C. albicans, C. glabrata, C. tropicalis, C. parapsilosis, C. lusitaniae, and C. kefyr. For each of the acquired strains, biofilm formation was evaluated on standardized samples of silicone pads and acrylic resin. We assessed the pathogenicity of the strains by infecting G. mellonella animals with Candida strains and observing survival. Results The biofilm formation and pathogenicity in Galleria was similar between oral and systemic isolates. The quantity of biofilm formed and the virulence in G. mellonella were different for each of the species studied. On silicone pads, C. albicans and C. dubliniensis produced more biofilm (1.12 to 6.61 mg than the other species (0.25 to 3.66 mg. However, all Candida species produced a similar biofilm on acrylic resin, material used in dental prostheses. C. albicans, C. dubliniensis, C. tropicalis, and C. parapsilosis were the most virulent species in G. mellonella with 100% of mortality, followed by C. lusitaniae (87%, C. novergensis (37%, C. krusei (25%, C. glabrata (20%, and C. kefyr (12%. Conclusions We found that on silicone pads as well as in the Galleria model, biofilm formation and virulence depends on the Candida species. Importantly, for C. albicans the pathogenicity of oral Candida isolates was similar to systemic Candida isolates

  17. Whole genome typing of the recently emerged Canadian serogroup W Neisseria meningitidis sequence type 11 clonal complex isolates associated with invasive meningococcal disease

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    Raymond S.W. Tsang

    2018-04-01

    Full Text Available Objectives: This study was performed to analyze the Canadian invasive serogroup W Neisseria meningitidis (MenW sequence type 11 (ST-11 clonal complex (CC isolates by whole genome typing and to compare Canadian isolates with similar isolates from elsewhere. Methods: Whole genome typing of 30 MenW ST-11 CC, 20 meningococcal group C (MenC ST-11 CC, and 31 MenW ST-22 CC isolates was performed on the Bacterial Isolate Genome Sequence database platform. Canadian MenW ST-11 CC isolates were compared with the 2000 MenW Hajj outbreak strain, as well as with MenW ST-11 CC from other countries. Results: Whole genome typing showed that the Canadian MenW ST-11 CC isolates were distinct from the traditional MenW ST-22 CC; they were not capsule-switched contemporary MenC strains that incorporated MenW capsules. While some recent MenW disease cases in Canada were caused by MenW ST-11 CC isolates showing relatedness to the 2000 MenW Hajj strain, many were non-Hajj isolates similar to current MenW ST-11 isolates found globally. Geographical and temporal variations in genotypes and surface protein antigen genes were found among the MenW ST-11 CC isolates. Conclusions: The current MenW ST-11 isolates did not arise by capsule switching from contemporary MenC ST-11 isolates. Both the Hajj-related and non-Hajj MenW ST-11 CC strains were associated with invasive meningococcal disease in Canada. Keywords: Neisseria meningitidis, Invasive meningococcal disease, Whole genome typing

  18. Molecular Analysis of Rising Fluoroquinolone Resistance in Belgian Non-Invasive Streptococcus pneumoniae Isolates (1995-2014)

    Science.gov (United States)

    Ceyssens, Pieter-Jan; Van Bambeke, Françoise; Mattheus, Wesley; Bertrand, Sophie; Fux, Frédéric; Van Bossuyt, Eddie; Damée, Sabrina; Nyssen, Henry-Jean; De Craeye, Stéphane; Verhaegen, Jan; Tulkens, Paul M.; Vanhoof, Raymond

    2016-01-01

    We present the results of a longitudinal surveillance study (1995–2014) on fluoroquinolone resistance (FQ-R) among Belgian non-invasive Streptococcus pneumoniae isolates (n = 5,602). For many years, the switch to respiratory fluoroquinolones for the treatment of (a)typical pneumonia had no impact on FQ-R levels. However, since 2011 we observed a significant decrease in susceptibility towards ciprofloxacin, ofloxacin and levofloxacin with peaks of 9.0%, 6.6% and 3.1% resistant isolates, respectively. Resistance to moxifloxacin arised sporadically, and remained <1% throughout the entire study period. We observed classical topoisomerase mutations in gyrA (n = 25), parC (n = 46) and parE (n = 3) in varying combinations, arguing against clonal expansion of FQ-R. The impact of recombination with co-habiting commensal streptococci on FQ-R remains marginal (10.4%). Notably, we observed that a rare combination of DNA Gyrase mutations (GyrA_S81L/GyrB_P454S) suffices for high-level moxifloxacin resistance, contrasting current model. Interestingly, 85/422 pneumococcal strains display MICCIP values which were lowered by at least four dilutions by reserpine, pointing at involvement of efflux pumps in FQ-R. In contrast to susceptible strains, isolates resistant to ciprofloxacin significantly overexpressed the ABC pump PatAB in comparison to reference strain S. pneumoniae ATCC 49619, but this could only be linked to disruptive terminator mutations in a fraction of these. Conversely, no difference in expression of the Major Facilitator PmrA, unaffected by reserpine, was noted between susceptible and resistant S. pneumoniae strains. Finally, we observed that four isolates displayed intermediate to high-level ciprofloxacin resistance without any known molecular resistance mechanism. Focusing future molecular studies on these isolates, which are also commonly found in other studies, might greatly assist in the battle against rising pneumococcal drug resistance. PMID:27227336

  19. Molecular Analysis of Rising Fluoroquinolone Resistance in Belgian Non-Invasive Streptococcus pneumoniae Isolates (1995-2014.

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    Pieter-Jan Ceyssens

    Full Text Available We present the results of a longitudinal surveillance study (1995-2014 on fluoroquinolone resistance (FQ-R among Belgian non-invasive Streptococcus pneumoniae isolates (n = 5,602. For many years, the switch to respiratory fluoroquinolones for the treatment of (atypical pneumonia had no impact on FQ-R levels. However, since 2011 we observed a significant decrease in susceptibility towards ciprofloxacin, ofloxacin and levofloxacin with peaks of 9.0%, 6.6% and 3.1% resistant isolates, respectively. Resistance to moxifloxacin arised sporadically, and remained <1% throughout the entire study period. We observed classical topoisomerase mutations in gyrA (n = 25, parC (n = 46 and parE (n = 3 in varying combinations, arguing against clonal expansion of FQ-R. The impact of recombination with co-habiting commensal streptococci on FQ-R remains marginal (10.4%. Notably, we observed that a rare combination of DNA Gyrase mutations (GyrA_S81L/GyrB_P454S suffices for high-level moxifloxacin resistance, contrasting current model. Interestingly, 85/422 pneumococcal strains display MICCIP values which were lowered by at least four dilutions by reserpine, pointing at involvement of efflux pumps in FQ-R. In contrast to susceptible strains, isolates resistant to ciprofloxacin significantly overexpressed the ABC pump PatAB in comparison to reference strain S. pneumoniae ATCC 49619, but this could only be linked to disruptive terminator mutations in a fraction of these. Conversely, no difference in expression of the Major Facilitator PmrA, unaffected by reserpine, was noted between susceptible and resistant S. pneumoniae strains. Finally, we observed that four isolates displayed intermediate to high-level ciprofloxacin resistance without any known molecular resistance mechanism. Focusing future molecular studies on these isolates, which are also commonly found in other studies, might greatly assist in the battle against rising pneumococcal drug resistance.

  20. Molecular Analysis of Rising Fluoroquinolone Resistance in Belgian Non-Invasive Streptococcus pneumoniae Isolates (1995-2014).

    Science.gov (United States)

    Ceyssens, Pieter-Jan; Van Bambeke, Françoise; Mattheus, Wesley; Bertrand, Sophie; Fux, Frédéric; Van Bossuyt, Eddie; Damée, Sabrina; Nyssen, Henry-Jean; De Craeye, Stéphane; Verhaegen, Jan; Tulkens, Paul M; Vanhoof, Raymond

    2016-01-01

    We present the results of a longitudinal surveillance study (1995-2014) on fluoroquinolone resistance (FQ-R) among Belgian non-invasive Streptococcus pneumoniae isolates (n = 5,602). For many years, the switch to respiratory fluoroquinolones for the treatment of (a)typical pneumonia had no impact on FQ-R levels. However, since 2011 we observed a significant decrease in susceptibility towards ciprofloxacin, ofloxacin and levofloxacin with peaks of 9.0%, 6.6% and 3.1% resistant isolates, respectively. Resistance to moxifloxacin arised sporadically, and remained topoisomerase mutations in gyrA (n = 25), parC (n = 46) and parE (n = 3) in varying combinations, arguing against clonal expansion of FQ-R. The impact of recombination with co-habiting commensal streptococci on FQ-R remains marginal (10.4%). Notably, we observed that a rare combination of DNA Gyrase mutations (GyrA_S81L/GyrB_P454S) suffices for high-level moxifloxacin resistance, contrasting current model. Interestingly, 85/422 pneumococcal strains display MICCIP values which were lowered by at least four dilutions by reserpine, pointing at involvement of efflux pumps in FQ-R. In contrast to susceptible strains, isolates resistant to ciprofloxacin significantly overexpressed the ABC pump PatAB in comparison to reference strain S. pneumoniae ATCC 49619, but this could only be linked to disruptive terminator mutations in a fraction of these. Conversely, no difference in expression of the Major Facilitator PmrA, unaffected by reserpine, was noted between susceptible and resistant S. pneumoniae strains. Finally, we observed that four isolates displayed intermediate to high-level ciprofloxacin resistance without any known molecular resistance mechanism. Focusing future molecular studies on these isolates, which are also commonly found in other studies, might greatly assist in the battle against rising pneumococcal drug resistance.

  1. Improved In Vitro Culture of Plasmodium falciparum Permits Establishment of Clinical Isolates with Preserved Multiplication, Invasion and Rosetting Phenotypes

    Science.gov (United States)

    Albrecht, Letusa; Ahmed Ismail, Hodan; Normark, Johan; Flaberg, Emilie; Szekely, Laszlo; Hultenby, Kjell; Persson, Kristina E. M.; Egwang, Thomas G.; Wahlgren, Mats

    2013-01-01

    To be able to robustly propagate P. falciparum at optimal conditions in vitro is of fundamental importance for genotypic and phenotypic studies of both established and fresh clinical isolates. Cryo-preserved P. falciparum isolates from Ugandan children with severe or uncomplicated malaria were investigated for parasite phenotypes under different in vitro growth conditions or studied directly from the peripheral blood. The parasite cultures showed a minimal loss of parasite-mass and preserved percentage of multiple infected pRBCs to that in peripheral blood, maintained adhesive phenotypes and good outgrowth and multiplication rates when grown in suspension and supplemented with gas. In contrast, abnormal and greatly fluctuating levels of multiple infections were observed during static growth conditions and outgrowth and multiplication rates were inferior. Serum, as compared to Albumax, was found necessary for optimal presentation of PfEMP1 at the pRBC surface and/or for binding of serum proteins (immunoglobulins). Optimal in vitro growth conditions of P. falciparum therefore include orbital shaking (50 rev/min), human serum (10%) and a fixed gas composition (5% O2, 5% CO2, 90% N2). We subsequently established 100% of 76 frozen patient isolates and found rosetting with schizont pRBCs in every isolate (>26% schizont rosetting rate). Rosetting during schizogony was often followed by invasion of the bound RBC as seen by regular and time-lapse microscopy as well as transmission electron microscopy. The peripheral parasitemia, the level of rosetting and the rate of multiplication correlated positively to one another for individual isolates. Rosetting was also more frequent with trophozoite and schizont pRBCs of children with severe versus uncomplicated malaria (p<0.002; p<0.004). The associations suggest that rosetting enhances the ability of the parasite to multiply within the human host. PMID:23894537

  2. Molecular analyses of Fusarium isolates recovered from a cluster of invasive mold infections in a Brazilian hospital

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    Scheel Christina M

    2013-01-01

    Full Text Available Abstract Background Invasive fusariosis (IF is a rare but often fatal fungal infection in immunosuppressed patients. In 2007, cases of IF above the expected epidemiologic baseline were detected in the hematology ward of a hospital in Rio de Janeiro, Brazil. Possible sources of infection were investigated by performing environmental sampling and patient isolate collection, followed by molecular typing. Isolates from dermatology patients with superficial fusariosis were included in the study for comparison to molecular types found in the community. Methods Environmental sampling focused on water-related sources in and around the hematology ward. Initially, we characterized 166 clinical and environmental isolates using the Fusarium translation elongation factor 1α (EF-1α genetic locus. Isolates included 68 collected from water-related sources in the hospital environment, 55 from 18 hematology patients, and 43 from the skin/nails of 40 outpatients seen at the hospital dermatology clinic. Multi-locus sequence typing was performed on Fusarium solani species complex (FSSC species 1 and 2 isolates to investigate their relatedness further. Results Most of the hematology samples were FSSC species 2, with species type FSSC 2-d the most commonly isolated from these patients. Most of the outpatient dermatology samples were also FSSC 2, with type 2-d again predominating. In contrast, environmental isolates from water sources were mostly Fusarium oxysporum species complex (FOSC and those from air samples mostly Fusarium incarnatum-equiseti species complex (FIESC. A third of the environmental samples were FSSC, with species types FSSC 1-a and FSSC 1-b predominating. Conclusions Fusarium isolate species types from hematology patient infections were highly similar to those recovered from dermatology patients in the community. Four species types (FSSC 1-a, 1-b, 2-d and 2-f were shared between hematology patients and the environment. Limitations in environmental

  3. PCR detection of four virulence-associated genes of Campylobacter jejuni isolates from Thai broilers and their abilities of adhesion to and invasion of INT-407 cells.

    Science.gov (United States)

    Chansiripornchai, Niwat; Sasipreeyajan, Jiroj

    2009-06-01

    Campylobacter jejuni is a major cause of food borne pathogens in humans and a major reservoir for this pathogen is poultry. The C. jejuni in broilers was investigated from in the caeca of broilers. Twenty broiler/flock samples from 7 flocks were assessed. The average prevalence of C. jejuni was 65% in the broiler flocks. The adhesion and invasion ability of 48 strains of C. jejuni on INT 407 were studied. The adhesion and invasion ability of 48 Campylobacter isolates from caecal contents were analyzed with Human embryonic intestine (INT-407) cells being used as a gentamicin resistance assay. The caecal isolates exhibited a wide range of adherence and invasion ability. There was a significant correlation (pCampylobacter isolates. Each of the virulence-associated genes: dnaJ, cadF, pldA and ciaB was detected by polymerase chain reaction from 100, 76, 31 and 41% of the Campylobacter strains, respectively. All of four virulence-associated genes were detected in 11 isolates. However, there was unclear association between the invasion ability and the presence of virulence-associated genes in this experiment, suggesting that more genes may be involved in the invasion process.

  4. High frequency of silver resistance genes in invasive isolates of Enterobacter and Klebsiella species.

    Science.gov (United States)

    Sütterlin, S; Dahlö, M; Tellgren-Roth, C; Schaal, W; Melhus, Å

    2017-07-01

    Silver-based products have been marketed as an alternative to antibiotics, and their consumption has increased. Bacteria may, however, develop resistance to silver. To study the presence of genes encoding silver resistance (silE, silP, silS) over time in three clinically important Enterobacteriaceae genera. Using polymerase chain reaction (PCR), 752 bloodstream isolates from the years 1990-2010 were investigated. Age, gender, and ward of patients were registered, and the susceptibility to antibiotics and silver nitrate was tested. Clonality and single nucleotide polymorphism were assessed with repetitive element sequence-based PCR, multi-locus sequence typing, and whole-genome sequencing. Genes encoding silver resistance were detected most frequently in Enterobacter spp. (48%), followed by Klebsiella spp. (41%) and Escherichia coli 4%. Phenotypical resistance to silver nitrate was found in Enterobacter (13%) and Klebsiella (3%) isolates. The lowest carriage rate of sil genes was observed in blood isolates from the neonatology ward (24%), and the highest in blood isolates from the oncology/haematology wards (66%). Presence of sil genes was observed in international high-risk clones. Sequences of the sil and pco clusters indicated that a single mutational event in the silS gene could have caused the phenotypic resistance. Despite a restricted consumption of silver-based products in Swedish health care, silver resistance genes are widely represented in clinical isolates of Enterobacter and Klebsiella species. To avoid further selection and spread of silver-resistant bacteria with a high potential for healthcare-associated infections, the use of silver-based products needs to be controlled and the silver resistance monitored. Copyright © 2017 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

  5. Escherichia coli N-Acetylglucosamine-1-Phosphate-Uridyltransferase/Glucosamine-1-Phosphate-Acetyltransferase (GlmU) Inhibitory Activity of Terreic Acid Isolated from Aspergillus terreus.

    Science.gov (United States)

    Sharma, Rashmi; Lambu, Mallikharjuna Rao; Jamwal, Urmila; Rani, Chitra; Chib, Reena; Wazir, Priya; Mukherjee, Debaraj; Chaubey, Asha; Khan, Inshad Ali

    2016-04-01

    Secondary metabolite of Aspergillus terreus, terreic acid, is a reported potent antibacterial that was identified more than 60 years ago, but its cellular target(s) are still unknown. Here we screen its activity against the acetyltransferase domain of a bifunctional enzyme, Escherichia coli N-acetylglucosamine-1-phosphate-uridyltransferase/glucosamine-1-phosphate-acetyltransferase (GlmU). An absorbance-based assay was used to screen terreic acid against the acetyltransferase activity of E. coli GlmU. Terreic acid was found to inhibit the acetyltransferase domain of E. coli GlmU with an IC50 of 44.24 ± 1.85 µM. Mode of inhibition studies revealed that terreic acid was competitive with AcCoA and uncompetitive with GlcN-1-P. It also exhibited concentration-dependent killing of E. coli ATCC 25922 up to 4× minimum inhibitory concentration and inhibited the growth of biofilms generated by E. coli. Characterization of resistant mutants established mutation in the acetyltransferase domain of GlmU. Terreic acid was also found to be metabolically stable in the in vitro incubations with rat liver microsome in the presence of a NADPH regenerating system. The studies reported here suggest that terreic acid is a potent antimicrobial agent and support that E. coli GlmU acetyltransferase is a molecular target of terreic acid, resulting in its antibacterial activity. © 2016 Society for Laboratory Automation and Screening.

  6. Isolation, identification, and characterization of an Aspergillus niger bioflocculant-producing strain using potato starch wastewater as nutrilite and its application.

    Science.gov (United States)

    Pu, Shengyan; Ma, Hui; Deng, Daili; Xue, Shengyang; Zhu, Rongxin; Zhou, Yan; Xiong, Xingying

    2018-01-01

    A bioflocculant (MBFA18) was produced by Aspergillus niger (A18) using potato starch wastewater (PSW) as nutrients. The cultivation processes and flocculating treatment for PSW purification were systematically studied. The flocculating rate of the MBFA 18 achieved 90.06% (kaolin clay) under the optimal cultivation condition (PSW with 5950 mg/L COD, 20 g/L glucose, 0.2 g/L urea and without phosphorus source addition and pH adjustment). Furthermore, effects of flocculant dosage, initial pH, coagulant aid (CaCl2) addition and sedimentation time on the PSW treatment were discussed and studied in detail. The optimum flocculation treatment conditions were determined according to the treatment efficiency, cost and flocculation conditions. During the PSW treatment, 2 mL/L bioflocculant (1.89 g/L) dosage and 0.5 mol/L coagulant aid addition were applied without pH adjustment and 91.15% COD and 60.22% turbidity removal rate could be achieved within 20 min. The comparative study between the bioflocculant and conventional chemical flocculants showed excellent flocculating efficiency of MBFA 18 with lower cost (4.7 yuan/t), which indicated that the bioflocculant MBFA 18 produced in PSW substrate has a great potential to be an alternative flocculant in PSW treatment.

  7. Invasive bacterial disease trends and characterization of group B streptococcal isolates among young infants in southern Mozambique, 2001-2015.

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    Betuel Sigaúque

    Full Text Available Maternal group B streptococcal (GBS vaccines under development hold promise to prevent GBS disease in young infants. Sub-Saharan Africa has the highest estimated disease burden, although data on incidence and circulating strains are limited. We described invasive bacterial disease (IBD trends among infants <90 days in rural Mozambique during 2001-2015, with a focus on GBS epidemiology and strain characteristics.Community-level birth and mortality data were obtained from Manhiça's demographic surveillance system. IBD cases were captured through ongoing surveillance at Manhiça district hospital. Stored GBS isolates from cases underwent serotyping by multiplex PCR, antimicrobial susceptibility testing, and whole genome sequencing.There were 437 IBD cases, including 57 GBS cases. Significant declines in overall IBD, neonatal mortality, and stillbirth rates were observed (P<0.0001, but not for GBS (P = 0.17. In 2015, GBS was the leading cause of young infant IBD (2.7 per 1,000 live births. Among 35 GBS isolates available for testing, 31 (88.6% were highly related serotype III isolates within multilocus sequence types (STs 17 (68.6% or 109 (20.0%. All seven ST109 isolates (21.9% had elevated minimum inhibitory concentration (MIC to penicillin (≥0.12 μg/mL associated with penicillin-binding protein (PBP 2x substitution G398A. Epidemiologic and molecular data suggest this is a well-established clone.A notable young infant GBS disease burden persisted despite improvements in overall maternal and neonatal health. We report an established strain with pbp2x point mutation, a first-step mutation associated with reduced penicillin susceptibility within a well-known virulent lineage in rural Mozambique. Our findings further underscores the need for non-antibiotic GBS prevention strategies.

  8. Genetic diversity of pneumococcal surface protein A in invasive pneumococcal isolates from Korean children, 1991-2016.

    Directory of Open Access Journals (Sweden)

    Ki Wook Yun

    Full Text Available Pneumococcal surface protein A (PspA is an important virulence factor of pneumococci and has been investigated as a primary component of a capsular serotype-independent pneumococcal vaccine. Thus, we sought to determine the genetic diversity of PspA to explore its potential as a vaccine candidate. Among the 190 invasive pneumococcal isolates collected from Korean children between 1991 and 2016, two (1.1% isolates were found to have no pspA by multiple polymerase chain reactions. The full length pspA genes from 185 pneumococcal isolates were sequenced. The length of pspA varied, ranging from 1,719 to 2,301 base pairs with 55.7-100% nucleotide identity. Based on the sequences of the clade-defining regions, 68.7% and 49.7% were in PspA family 2 and clade 3/family 2, respectively. PspA clade types were correlated with genotypes using multilocus sequence typing and divided into several subclades based on diversity analysis of the N-terminal α-helical regions, which showed nucleotide sequence identities of 45.7-100% and amino acid sequence identities of 23.1-100%. Putative antigenicity plots were also diverse among individual clades and subclades. The differences in antigenicity patterns were concentrated within the N-terminal 120 amino acids. In conclusion, the N-terminal α-helical domain, which is known to be the major immunogenic portion of PspA, is genetically variable and should be further evaluated for antigenic differences and cross-reactivity between various PspA types from pneumococcal isolates.

  9. Genetic diversity of pneumococcal surface protein A in invasive pneumococcal isolates from Korean children, 1991-2016.

    Science.gov (United States)

    Yun, Ki Wook; Choi, Eun Hwa; Lee, Hoan Jong

    2017-01-01

    Pneumococcal surface protein A (PspA) is an important virulence factor of pneumococci and has been investigated as a primary component of a capsular serotype-independent pneumococcal vaccine. Thus, we sought to determine the genetic diversity of PspA to explore its potential as a vaccine candidate. Among the 190 invasive pneumococcal isolates collected from Korean children between 1991 and 2016, two (1.1%) isolates were found to have no pspA by multiple polymerase chain reactions. The full length pspA genes from 185 pneumococcal isolates were sequenced. The length of pspA varied, ranging from 1,719 to 2,301 base pairs with 55.7-100% nucleotide identity. Based on the sequences of the clade-defining regions, 68.7% and 49.7% were in PspA family 2 and clade 3/family 2, respectively. PspA clade types were correlated with genotypes using multilocus sequence typing and divided into several subclades based on diversity analysis of the N-terminal α-helical regions, which showed nucleotide sequence identities of 45.7-100% and amino acid sequence identities of 23.1-100%. Putative antigenicity plots were also diverse among individual clades and subclades. The differences in antigenicity patterns were concentrated within the N-terminal 120 amino acids. In conclusion, the N-terminal α-helical domain, which is known to be the major immunogenic portion of PspA, is genetically variable and should be further evaluated for antigenic differences and cross-reactivity between various PspA types from pneumococcal isolates.

  10. Metabolomics Analysis Reveals Specific Novel Tetrapeptide and Potential Anti-Inflammatory Metabolites in Pathogenic Aspergillus species

    Directory of Open Access Journals (Sweden)

    Kim-Chung Lee

    2015-06-01

    Full Text Available Infections related to Aspergillus species have emerged to become an important focus in infectious diseases, as a result of the increasing use of immunosuppressive agents and high fatality associated with invasive aspergillosis. However, laboratory diagnosis of Aspergillus infections remains difficult. In this study, by comparing the metabolomic profiles of the culture supernatants of 30 strains of six pathogenic Aspergillus species (A. fumigatus, A. flavus, A. niger, A. terreus, A. nomius and A. tamarii and 31 strains of 10 non-Aspergillus fungi, eight compounds present in all strains of the six Aspergillus species but not in any strain of the non-Aspergillus fungi were observed. One of the eight compounds, Leu–Glu–Leu–Glu, is a novel tetrapeptide and represents the first linear tetrapeptide observed in Aspergillus species, which we propose to be named aspergitide. Two other closely related Aspergillus-specific compounds, hydroxy-(sulfooxybenzoic acid and (sulfooxybenzoic acid, may possess anti-inflammatory properties, as 2-(sulfooxybenzoic acid possesses a structure similar to those of aspirin [2-(acetoxybenzoic acid] and salicylic acid (2-hydroxybenzoic acid. Further studies to examine the potentials of these Aspergillus-specific compounds for laboratory diagnosis of aspergillosis are warranted and further experiments will reveal whether Leu–Glu–Leu–Glu, hydroxy-(sulfooxybenzoic acid and (sulfooxybenzoic acid are virulent factors of the pathogenic Aspergillus species.

  11. Exposure to an environmental estrogen breaks down sexual isolation between native and invasive species.

    Science.gov (United States)

    Ward, Jessica L; Blum, Michael J

    2012-12-01

    Environmental change can increase the likelihood of interspecific hybridization by altering properties of mate recognition and discrimination between sympatric congeners. We examined how exposure to an environmentally widespread endocrine-disrupting chemical (EDC), bisphenol A (BPA), affected visual communication signals and behavioral isolation between an introduced freshwater fish and a native congener (genus: Cyprinella). Exposure to BPA induced changes in the expression of male secondary traits as well as male and female mate choice, leading to an overall reduction in prezygotic isolation between congeners. Changes in female mate discrimination were not tightly linked to changes in male phenotypic traits, suggesting that EDC exposure may alter female choice thresholds independently of the effects of exposure on males. These findings indicate that environmental exposure to EDCs can lead to population declines via the erosion of species boundaries and by promoting the establishment and spread of non-native species via hybridization.

  12. Animal Models for Studying Triazole Resistance in Aspergillus fumigatus

    NARCIS (Netherlands)

    Lewis, R.E.; Verweij, P.E.

    2017-01-01

    Infections caused by triazole-resistant Aspergillus fumigatus are associated with a higher probability of treatment failure and mortality. Because clinical experience in managing these infections is still limited, mouse models of invasive aspergillosis fulfill a critical void for studying treatment

  13. Pharmacodynamics of isavuconazole in an Aspergillus fumigatus mouse infection model

    NARCIS (Netherlands)

    Seyedmousavi, S.; Bruggemann, R.J.; Meis, J.F.G.M.; Melchers, W.J.G.; Verweij, P.E.; Mouton, J.W.

    2015-01-01

    Azole resistance is an emerging problem in Aspergillus fumigatus which translates into treatment failure. Alternative treatments with new azoles may improve therapeutic outcome in invasive aspergillosis (IA) even for strains with decreased susceptibility to current azoles. The in vivo efficacy of

  14. Emergence of azole resistance in Aspergillus fumigatus and spread of a single resistance mechanism.

    Directory of Open Access Journals (Sweden)

    Eveline Snelders

    2008-11-01

    Full Text Available BACKGROUND: Resistance to triazoles was recently reported in Aspergillus fumigatus isolates cultured from patients with invasive aspergillosis. The prevalence of azole resistance in A. fumigatus is unknown. We investigated the prevalence and spread of azole resistance using our culture collection that contained A. fumigatus isolates collected between 1994 and 2007. METHODS AND FINDINGS: We investigated the prevalence of itraconazole (ITZ resistance in 1,912 clinical A. fumigatus isolates collected from 1,219 patients in our University Medical Centre over a 14-y period. The spread of resistance was investigated by analyzing 147 A. fumigatus isolates from 101 patients, from 28 other medical centres in The Netherlands and 317 isolates from six other countries. The isolates were characterized using phenotypic and molecular methods. The electronic patient files were used to determine the underlying conditions of the patients and the presence of invasive aspergillosis. ITZ-resistant isolates were found in 32 of 1,219 patients. All cases were observed after 1999 with an annual prevalence of 1.7% to 6%. The ITZ-resistant isolates also showed elevated minimum inhibitory concentrations of voriconazole, ravuconazole, and posaconazole. A substitution of leucine 98 for histidine in the cyp51A gene, together with two copies of a 34-bp sequence in tandem in the gene promoter (TR/L98H, was found to be the dominant resistance mechanism. Microsatellite analysis indicated that the ITZ-resistant isolates were genetically distinct but clustered. The ITZ-sensitive isolates were not more likely to be responsible for invasive aspergillosis than the ITZ-resistant isolates. ITZ resistance was found in isolates from 13 patients (12.8% from nine other medical centres in The Netherlands, of which 69% harboured the TR/L98H substitution, and in six isolates originating from four other countries. CONCLUSIONS: Azole resistance has emerged in A. fumigatus and might be more

  15. Purification and Characterization of a Polyextremophilic α-Amylase from an Obligate Halophilic Aspergillus penicillioides Isolate and Its Potential for Souse with Detergents

    Directory of Open Access Journals (Sweden)

    Imran Ali

    2015-01-01

    Full Text Available An extracellular α-amylase from the obligate halophilic Aspergillus penicillioides TISTR3639 strain was produced and enriched to apparent homogeneity by ammonium sulfate precipitation and Sephadex G100 gel filtration column chromatography. The mass of the purified amylase was estimated to be 42 kDa by SDS-PAGE. With soluble starch as the substrate it had a specific activity of 118.42 U·mg−1 and Vmax⁡ and Km values of 1.05 µmol·min−1·mg−1 and 5.41 mg·mL−1, respectively. The enzyme was found to have certain polyextremophilic characteristics, with an optimum activity at pH 9, 80°C, and 300 g·L−1 NaCl. The addition of CaCl2 at 2 mM was found to slightly enhance the amylase activity, while ZnCl2, FeCl2, or EDTA at 2 mM was strongly or moderately inhibitory, respectively, suggesting the requirement for a (non-Fe2+ or Zn2+ divalent cation. The enzyme retained more than 80% of its activity when incubated with three different laundry detergents and had a better performance compared to a commercial amylase and three detergents in the presence of increasing NaCl concentrations up to 300 g·L−1. Accordingly, it has a good potential for use as an α-amylase in a low water activity (high salt concentration and at high pH and temperatures.

  16. Invasion and translocation of uropathogenic Escherichia coli isolated from urosepsis and patients with community-acquired urinary tract infection.

    Science.gov (United States)

    Owrangi, B; Masters, N; Kuballa, A; O'Dea, C; Vollmerhausen, T L; Katouli, M

    2018-01-16

    Uropathogenic Escherichia coli (UPEC) strains are found in high numbers in the gut of patients with urinary tract infections (UTIs). We hypothesised that in hospitalised patients, UPEC strains might translocate from the gut to the blood stream and that this could be due to the presence of virulence genes (VGs) that are not commonly found in UPEC strains that cause UTI only. To test this, E. coli strains representing 75 dominant clonal groups of UPEC isolated from the blood of hospitalised patients with UTI (urosepsis) (n = 22), hospital-acquired (HA) UTI without blood infection (n = 24) and strains isolated from patients with community-acquired (CA)-UTIs (n = 29) were tested for their adhesion to, invasion and translocation through Caco-2 cells, in addition to the presence of 34 VGs associated with UPEC. Although there were no differences in the rate and degree of translocation among the groups, urosepsis and HA-UTI strains showed significantly higher abilities to adhere (P = 0.0095 and P UPEC strains, irrespective of their source, are capable of translocating through gut epithelium. However, urosepsis and HA-UTI strains have a much better ability to interact with gut epithelia and have a greater virulence potential than CA-UPEC, which allows them to cause blood infection.

  17. A Rare Cause of Headache: Aspergillus Sinusitis

    Directory of Open Access Journals (Sweden)

    Şehnaz Arıcı

    2015-09-01

    Full Text Available Fungal sinusitis are mostly seen in immunosuppressive individuals and somtimes which can be mortal. Most frequently species of Aspergillus were isolated from, clinical forms of mycotic sinonasal disease.Surgical debridement,sinus ventilation and medical therapy in treatment of fungal sinusitis, are recommended. In this article, a case of healthy immune patient with fungal sinusitis who peresent with headache was repoted.

  18. Aflatoxin B 1 producing potential of Aspergillus flavus strains ...

    African Journals Online (AJOL)

    Aflatoxin B 1 (AFB1) producing potential of different strains of Aspergillus flavus, isolated from 1,200 stored rice grains collected from 43 locations in 20 rice growing states in India was investigated. Eighty-five strains of A. flavus were isolated from the discolored rice grains and tested for their AFB1 producing potential on ...

  19. Heavy metal biosorption potential of Aspergillus and Rhizopus sp ...

    African Journals Online (AJOL)

    Two isolates belonging to the predominant genera Aspergillus and Rhizopus isolated from agricultural field treated with sewage/ industrial effluents were selected for the biosorption potential evaluation of Cr and Cd. Pretreated, dead biomass of above fungi was used for bioadsorption experiment at pH value 4.5 with the ...

  20. Ochratoxin A production by strains of Aspergillus niger var. niger.

    Science.gov (United States)

    Abarca, M L; Bragulat, M R; Castellá, G; Cabañes, F J

    1994-01-01

    In a survey of the occurrence of ochratoxin A (OA)-positive strains isolated from feedstuffs, two of the 19 isolates of Aspergillus niger var. niger that were studied produced OA in 2% yeast extract-15% sucrose broth and in corn cultures. This is the first report of production of OA by this species. PMID:8074536

  1. Dynamics in prophage content of invasive and noninvasive M1 and M28 Streptococcus pyogenes isolates in the Netherlands from 1959 to 1996

    NARCIS (Netherlands)

    Vlaminckx, B.J.M.; Schuren, F.H.J.; Montijn, R.C.; Caspers, M.P.M.; Beitsma, M.M.; Wannet, W.J.B.; Schouls, L.M.; Verhoef, J.; Jansen, W.T.M.

    2007-01-01

    Invasive group A streptococcal (GAS) disease reemerged in The Netherlands in the late 1980s. To seek an explanation for this resurgence, the genetic compositions of 22 M1 and 19 M28 GAS strains isolated in The Netherlands between 1960s and the mid-1990s were analyzed by using a mixed-genome DNA

  2. Interspecies Recombination in Type II Topoisomerase Genes Is Not a Major Cause of Fluoroquinolone Resistance in Invasive Streptococcus pneumoniae Isolates in the United States

    OpenAIRE

    Pletz, Mathias W. R.; McGee, Lesley; Beall, Bernard; Whitney, Cynthia G.; Klugman, Keith P.

    2005-01-01

    Mutations in the topoisomerase type II enzymes account for fluoroquinolone resistance in Streptococcus pneumoniae. These mutations can arise spontaneously or be transferred by intraspecies or interspecies recombination, primarily with viridans streptococci. We analyzed the nucleotide sequences of the quinolone resistance-determining regions of 49 invasive levofloxacin-resistant pneumococcal isolates and did not find any evidence for interspecies recombination.

  3. Two-stage statistical medium optimization for augmented cellulase production via solid-state fermentation by newly isolated Aspergillus niger HN-1 and application of crude cellulase consortium in hydrolysis of rice straw.

    Science.gov (United States)

    Sandhu, Simranjeet Kaur; Oberoi, Harinder Singh; Babbar, Neha; Miglani, Kanupriya; Chadha, Bhupinder Singh; Nanda, Dhiraj Kumar

    2013-12-26

    Cellulolytic enzyme production by newly isolated Aspergillus niger HN-1 was statistically optimized using Plackett-Burman and central composite design (CCD). Optimum concentrations of 2, 0.40, 0.01, and 0.60 g L (-1) for KH2PO4, urea, trace elements solution, and CaCl2·2H2O, respectively, were suggested by Design-Expert software. The two-stage optimization process led to a 3- and 2-fold increases in the filter paper cellulase (FP) and β-glucosidase activities, respectively. FP, β-glucosidase, endoglucanase, exopolygalaturonase, cellobiohydrolase, xylanase, α-l-arabinofuranosidase, β-xylosidase, and xylan esterase activities of 36.7 ± 1.54 FPU gds(-1), 252.3 ± 7.4 IU gds(-1), 416.3 ± 22.8 IU gds(-1), 111.2 ± 5.4 IU gds(-1), 8.9 ± 0.50 IU gds(-1), 2593.5 ± 78.9 IU gds(-1), 79.4 ± 4.3 IU gds(-1), 180.8 ± 9.3 IU gds(-1), and 288.7 ± 11.8 IU gds(-1), respectively, were obtained through solid-state fermentation during the validation studies. Hydrolysis of alkali-treated rice straw with crude cellulases resulted in about 84% glucan to glucose, 89% xylan to xylose, and 91% arabinan to arabinose conversions, indicating potential for biomass hydrolysis by the crude cellulase consortium obtained in this study.

  4. Identification and antifungal activity of Streptomyces sp. S72 isolated ...

    African Journals Online (AJOL)

    The test of antifungal activity for several pathogens fungi causing invasive aspergillosis and systemic candidiasis revealed that the Streptomyces sp. S72 was a good moderate antifungal compound producer against Aspergillus fumigatus and Candida albicans, and had no activity against Aspergillus flavus, Aspergillus ...

  5. Evaluation of the pathogenic effect of some fungal isolates on fruits ...

    African Journals Online (AJOL)

    PDA). The consistently isolated fungi from the fruits and seeds were identified as Penicillium digitatum, Aspergillus flavus and Aspergillus niger while a specie of Verticillium was isolated from the leaves and stems. Pathogenicity tests on fresh ...

  6. Potential of syncytiotrophoblasts isolated from the cervical mucus for early non-invasive prenatal diagnosis: evidence of a vanishing twin.

    Science.gov (United States)

    Mantzaris, Debbie; Cram, David S

    2015-01-01

    Non-invasive methods to assess the foetal genome during pregnancy will provide new opportunities to offer pregnant women a more comprehensive genetic diagnosis of their established foetus. The aim of this study was to determine the presence and frequency of foetal cells in transcervical cell (TCC) mucus samples from pregnant women and determine their suitability for early prenatal diagnosis. Syncytiotrophoblasts in aspirated TCC mucus samples were identified by immunostaining with the foetal-specific antibody NDOG1. Genetic analysis of foetal cells was performed by laser capture microdissection and quantitative fluorescent PCR (QF-PCR). In 116 of 207 (56%) TCC samples, abundant syncytiotrophoblasts were retrieved. However, when TCC samples were stratified for the presence of chorionic villous fragments, syncytiotrophoblasts were identified in 85 of 109 (78%) samples. Significant numbers of syncytiotrophoblasts were found in TCC samples collected between 6 and 9weeks of gestation (mean 741, range 25-2884). QF-PCR analysis of NDOG1 positive syncytiotrophoblasts and matching maternal DNA confirmed their foetal origin and correct foetal cell sexing was achieved in 97% of TCC samples. The one discordant sex diagnosis was associated with a dizygotic dichorionic twin pregnancy resulting from the implantation of a female T21 embryo and a normal male embryo, where the female T21 foetus had succumbed at 6weeks of gestation and was vanishing. Syncytiotrophoblasts can be successfully isolated from TCC samples and represent a suitable source of cells for genetic analysis of the established foetus in early pregnancy. The study highlights a vanishing twin as a potential cause for discordant non-invasive prenatal test results. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Occurrence of toxigenic Aspergillus flavus in commercial Bulgur wheat

    Directory of Open Access Journals (Sweden)

    Carla Bertechini FARIA

    Full Text Available Abstract Aflatoxins are mutagenic, carcinogenic, and teratogenic mycotoxins. The objective of this work was to study the presence of aflatoxigenic Aspergillus in commercial Bulgur wheat in the city of Maringá, Paraná, Brazil. Thirty samples of commercial Bulgur wheat, acquired in the period of August 2011 to January 2012, were evaluated. The enumeration analysis showed that samples had up to 273.3 CFU of molds and 133.3 CFU of aflatoxigenic Aspergillus per gram of wheat. Forty-two monosporic isolates were obtained and identified as Aspergillus flavus. The isolates were analyzed regarding their aflatoxigenic potential by culture in coconut milk agar; hydroxide vapor exposure; chromatography; and polymerase chain reaction (PCR targeting genes that code enzymes of the aflatoxins synthesis pathway. Some of the isolates were confirmed to be aflatoxin producers and several of them presented a genetic profile of aflatoxin synthesis. The obtained results demonstrated that Bulgur wheat A. flavus contamination is concerning.

  8. Sinusite invasiva por Aspergillus flavus : relato de um caso associado a leucemia aguda bifenotípica

    OpenAIRE

    Xavier, Melissa Orzechowski; Oliveira, Flávio de Mattos; Almeida, Valdir de; Prolla, Gabriel; Severo, Luiz Carlos

    2009-01-01

    Here we report a case of invasive pansinusitis with proptosis of the right eye caused by Aspergillus flavus in an immunocompromised patient with acute biphenotypic leukemia without aggressive therapy response.Descreve-se um caso de pansinusite invasiva com proptose do globo ocular direito causado por Aspergillus flavus em um paciente imunossuprimido com leucemia aguda bifenotípica sem resposta a terapia agressiva.

  9. New ochratoxin A or sclerotium producing species in Aspergillus section Nigri

    DEFF Research Database (Denmark)

    Samson, R.A.; Frisvad, Jens Christian

    2004-01-01

    in Costa Rica and produces large pink to greyish brown sclerotia. Aspergillus lacticoffeatus was found on coffee beans in Venezuela and Indonesia, and is an effective producer of ochratoxin A. Aspergillus piperis was isolated from black ground pepper and produces large yellow to pink brown sclerotia...

  10. Spectrophotometric reading of EUCAST antifungal susceptibility testing of Aspergillus fumigatus

    DEFF Research Database (Denmark)

    Meletiadis, J.; Leth Mortensen, K.; Verweij, P. E.

    2017-01-01

    Objectives Given the increasing number of antifungal drugs and the emergence of resistant Aspergillus isolates, objective, automated and high-throughput antifungal susceptibility testing is important. The EUCAST E.Def 9.3 reference method for MIC determination of Aspergillus species relies on vis...

  11. High prevalence of azole-resistant Aspergillus fumigatus in adults with cystic fibrosis exposed to itraconazole.

    NARCIS (Netherlands)

    Burgel, P.R.; Baixench, M.T.; Amsellem, M.; Audureau, E.; Chapron, J.; Kanaan, R.; Honore, I.; Dupouy-Camet, J.; Dusser, D.; Klaassen, C.H.; Meis, J.F.G.M.; Hubert, D.; Paugam, A.

    2012-01-01

    Aspergillus fumigatus is the most frequent fungus found in the sputum of cystic fibrosis (CF) subjects. Itraconazole is prescribed for allergic bronchopulmonary aspergillosis (ABPA) or Aspergillus bronchitis in CF subjects. We hypothesized that A. fumigatus isolates in the sputum of CF subjects with

  12. Phenotypic differentiation of species from Aspergillus section Flavi on neutral red desiccated coconut agar

    DEFF Research Database (Denmark)

    Atanda, O. O.; Adetunji, M. C.; Ezekiel, C. N.

    2014-01-01

    In order to facilitate easy and rapid identification of aflatoxin-producing Aspergillus species, the phenotypic traits of Aspergillus section Flavi isolates were examined on neutral red desiccated coconut agar (NRDCA). Phenotype variations in colony morphology and the relationship between colour...

  13. Invasive aspergillosis: epidemiology and environmental study in haematology patients (Sfax, Tunisia).

    Science.gov (United States)

    Hadrich, I; Makni, F; Sellami, H; Cheikhrouhou, F; Sellami, A; Bouaziz, H; Hdiji, S; Elloumi, M; Ayadi, A

    2010-09-01

    Invasive aspergillosis (IA) is a major opportunistic infection in haematology patients. Preventive measures are important to control IA because diagnosis is difficult and the outcome of treatment is poor. We prospectively examined the environmental contamination by Aspergillus and other fungal species and evaluated the prevalence of invasive aspergillosis in the protect unit of haematology. A three-year prospective study (December 2004-September 2007) was carried out in the department of haematology of Hedi Chaker Hospital. Suspected invasive aspergillosis cases were reviewed and classified as proven, probable and possible invasive aspergillosis using the EORTC criteria. During the study period, we collected weekly environmental samples (patient's rooms, tables and acclimatisers) and clinical samples from each patient (nasal, expectoration and auricular). Among 105 neutropenic patients, 16 had probable and 13 had possible IA. A total of 1680 clinical samples were collected and A. flavus was most frequently isolated (79.2%). Analysis of 690 environmental samples revealed that Penicillium (44%) was the most frequent followed by Cladosporium (20%), Aspergillus spp. (18%) and Alternaria (13%). The PCR-sequencing of 30 A. flavus isolates detected from clinical and environmental samples confirmed the mycological identification. Our findings underline the importance of environmental surveillance and strict application of preventive measures.

  14. mutant of Aspergillus niger

    African Journals Online (AJOL)

    STORAGESEVER

    2009-01-19

    Jan 19, 2009 ... enzyme glucose oxidase production from mutagenization of A. niger, resistant to a range of ... strain, forward from Department of Microbiology, was used and maintained on potato dextrose agar (PDA) slants, .... (1993). Induction of glucose oxidase, catalase and lactonase in. Aspergillus niger. Curr. Genet.

  15. Production of thermostable glucoamylase by newly isolated Aspergillus flavus A 1.1 and Thermomyces lanuginosus A 13.37 Produção e glucoamilase por Aspergillus flavus A1.1 e Thermomyces lanuginosus A13.37

    Directory of Open Access Journals (Sweden)

    Eleni Gomes

    2005-03-01

    Full Text Available Thirteen thermophilic fungal strains were isolated from agricultural soil, tubers and compost samples in tropical Brazil. Two strains were selected based on of their ability to produce considerable glucoamylase activity while growing in liquid medium at 45ºC with starch as the only carbon source. They were identified as Aspergillus flavus A1.1 and Thermomyces lanuginosus A 13.37 Tsiklinsky. The experiment to evaluate the effect of carbon source, temperature and initial pH of the medium on enzyme production was developed in a full factorial design (2x2x3. Enzyme productivity was influenced by the type of starch used as carbon source. Cassava starch showed to be a better substrate than corn starch for glucoamylase production by A. flavus but for T. lanuginosus the difference was not significant. Enzyme activities were determined using as substrates 0.3% soluble starch, 0.3% maltose or 0.3% of starch plus 0.1% maltose. The enzymes from A. flavus A1.1 hydrolyzed soluble starch preferentially but also exhibited a significant maltase activity. Moreover higher quantities of glucose were released when the substrate used was a mixture of starch and maltose, suggesting that this fungus produced two types of enzyme. In the case T. lanuginosus A 13.37, the substrate specificity test indicated that the enzyme released also hydrolyzed starch more efficiently than maltose, but there was no increase in the liberation of glucose when a mixture of starch and maltose was used as substrate, suggesting that only one type of enzyme was secreted. Glucoamylases produced from A. flavus A1.1 and T. lanuginous A.13-37 have high optimum temperature (65ºC and 70ºC and good thermostability in the absence of substrate (maintaining 50% of activity for 5 and 8 hours, respectively, at 60ºC and are stable over in a wide pH range. These new strains offer an attractive alternative source of enzymes for industrial starch processing.Entre 13 linhagens de fungos filamentosos

  16. Two novel aflatoxin-producing Aspergillus species from Argentinean peanuts

    DEFF Research Database (Denmark)

    Pildain, M.B.; Frisvad, Jens Christian; Vaamonde, G.

    2008-01-01

    (morphology and extrolite profiles) and molecular (beta-tubulin and calmodulin gene sequences) characters. A. minisclerotigenes resembles Aspergillus flavus and Aspergillus parvisclerotigenus in producing aflatoxins B-1 and B-2, cyclopiazonic acid, kojic acid and aspergillic acid, but in addition it produces...... aflatoxins G(1) and G(2), aflavarins, aflatrem, aflavinines, parasiticolides and paspaline. This species also includes several isolates previously assigned to A. flavus group II and three Australian soil isolates. A. arachidicola produces aflatoxins B-1, B-2, G(1) and G(2), kojic acid, chrysogine...

  17. [Aspergillus species in hospital environments with pediatric patients in critical condition].

    Science.gov (United States)

    Fernández, Mariana; Cattana, María; Rojas, Florencia; Sosa, María de Los Ángeles; Aguirre, Clarisa; Vergara, Marta; Giusiano, Gustavo

    2014-01-01

    Aspergillus is a group of opportunistic fungi that cause infections, with high morbimortality in immunosuppressed patients. Aspergillus fumigatus is the most frequent species in these infections, although the incidence of other species has increased in the last few years. To evaluate the air fungal load and the diversity of Aspergillus species in hospitals with pediatric patients in critical condition. The Intensive Care Unit and Burns Unit of a pediatric hospital were sampled every 15 days during the autumn and spring seasons. The air samples were collected with SAS Super 100(®) and the surface samples were collected by swab method. The UFC/m(3) counts found exceeded the acceptable levels. The UFC/m(3) and the diversity of Aspergillus species found in the Intensive Care Unit were higher than those found in the Burns Unit. The fungal load and the diversity of species within the units were higher than those in control environments. The use of both methods -SAS and swab- allowed the detection of a higher diversity of species, with 96 strains of Aspergillus being isolated and 12 species identified. The outstanding findings were Aspergillus sydowii, Aspergillus niger, Aspergillus flavus, Aspergillus terreus and Aspergillus parasiticus, due to their high frequency. Aspergillus fumigatus, considered unacceptable in indoor environments, was isolated in both units. Aspergillus was present with high frequency in these units. Several species are of interest in public health for being potential pathogenic agents. Air control and monitoring are essential in the prevention of these infections. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  18. Incidence of Cyp51 A key mutations in Aspergillus fumigatus-a study on primary clinical samples of immunocompromised patients in the period of 1995-2013.

    Directory of Open Access Journals (Sweden)

    Birgit Spiess

    Full Text Available As the incidence of azole resistance in Aspergillus fumigatus is rising and the diagnosis of invasive aspergillosis (IA in immunocompromised patients is rarely based on positive culture yield, we screened our Aspergillus DNA sample collection for the occurrence of azole resistance mediating cyp51 A key mutations. Using two established, a modified and a novel polymerase chain reaction (PCR assays followed by DNA sequence analysis to detect the most frequent mutations in the A. fumigatus cyp51 A gene conferring azole resistance (TR34 (tandem repeat, L98H and M220 alterations. We analyzed two itraconazole and voriconazole and two multi-azole resistant clinical isolates and screened 181 DNA aliquots derived from clinical samples (blood, bronchoalveolar lavage (BAL, biopsies, cerebrospinal fluid (CSF of 155 immunocompromised patients of our Aspergillus DNA sample collection, previously tested positive for Aspergillus DNA and collected between 1995 and 2013. Using a novel PCR assay for the detection of the cyp51 A 46 bp tandem repeat (TR46 directly from clinical samples, we found the alteration in a TR46/Y121F/T289A positive clinical isolate. Fifty stored DNA aliquots from clinical samples were TR46 negative. DNA sequence analysis revealed a single L98H mutation in 2010, two times the L98H alteration combined with TR34 in 2011 and 2012 and a so far unknown N90K mutation in 1998. In addition, four clinical isolates were tested positive for the TR34/L98H combination in the year 2012. We consider our assay of epidemiological relevance to detect A. fumigatus azole resistance in culture-negative clinical samples of immunocompromised patients; a prospective study is ongoing.

  19. Aspergillus bertholletius sp. nov. from Brazil nuts.

    Directory of Open Access Journals (Sweden)

    Marta H Taniwaki

    Full Text Available During a study on the mycobiota of brazil nuts (Bertholletia excelsa in Brazil, a new Aspergillus species, A. bertholletius, was found, and is described here. A polyphasic approach was applied using morphological characters, extrolite data as well as partial β-tubulin, calmodulin and ITS sequences to characterize this taxon. A. bertholletius is represented by nineteen isolates from samples of brazil nuts at various stages of production and soil close to Bertholletia excelsa trees. The following extrolites were produced by this species: aflavinin, cyclopiazonic acid, kojic acid, tenuazonic acid and ustilaginoidin C. Phylogenetic analysis using partial β-tubulin and camodulin gene sequences showed that A. bertholletius represents a new phylogenetic clade in Aspergillus section Flavi. The type strain of A. bertholletius is CCT 7615 ( = ITAL 270/06 = IBT 29228.

  20. Aspergillus triggers phenazine production in Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Jensen, Britt Guillaume; Jelsbak, Lars; Søndergaard, Ib

    Objectives: Pseudomonas aeruginosa is an opportunistic human pathogen, commonly infecting cystic fibrosis (CF) patients. Aspergilli, especially Aspergillus fumigatus, are also frequently isolated from CF patients. Our aim was to examine the possible interaction between P. aeruginosa and different...... in the contact area of A. niger, A. flavus, A. oryzae, but not A. fumigatus. In addition, other metabolites with UV chromophores similar to the phenazines were only found in the contact zone between Aspergillus and Pseudomonas. No change in secondary metabolite profiles were seen for the Aspergilli, when...... comparing with or without the presence of Pseudomonas. Conclusion: All Aspergilli tested, with the exception of A. fumigatus, triggered the upregulation of phenazine-1-carboxamide and phenazine-1-carboxylic acid production by P. aeruginosa. Surprisingly no changes in secondary metabolite profiles were...

  1. Molecular identification of fungal isolates and hatching success of green turtle (Chelonia mydas) nests.

    Science.gov (United States)

    Candan, Esra Deniz

    2018-02-26

    The aim of this study is to investigate the fungal diversity of green turtle nests and to examine phylogenetic relationships among these isolates. During the nesting season, samples of intra-nest sand and failed eggs were collected from 25% of the surviving nests in Sugözü Beaches, which are amongst the most important nesting beaches for endangered green turtles in the Mediterranean. Twenty-three fungi were identified by molecular techniques. Fungal isolates belonged to genera Aspergillus, Emericella, Rhizopus, Actinomucor and Apophysomyces with two undescribed species. Aspergillus variecolor, Aspergillus quadrilinieatus, Aspergillus tubingensis, Rhizopus oryzae, Actinomucor elegans and Apophysomyces variabilis were firstly detected in all sea turtle nests within this study. Our results demonstrate that 36.4% of the nests had fungal contamination. Also hatching success of the nests contaminated by fungi were significantly lower than the uncontaminated nests (P = 0.029). Also, this may represent a threat to marine turtles and a risk for the health of conservation workers. This study is the first molecular phylogenetic study associated with sea turtle nests in the eastern Mediterranean coast and contributes to the wider body of literature on fungal invasion of sea turtle nests with firstly isolated species. These findings are important for improving potential conservation measures for the nest sites.

  2. Ação fungitóxica do óleo essencial de Tanaecium nocturnum (Barb. Rodr. Bur. e K. Shum sobre o Aspergillus flavus isolado da castanha-do-Brasil (Bertholletia excelsa Fungitoxic action of the essential oil of Tanaecium nocturnum (Barb. Rodr. Bur. and K. Shum on Aspergillus flavus isolated from the Brazil nut (Bertholletia excelsa

    Directory of Open Access Journals (Sweden)

    Flávio Araújo Pimentel

    2010-03-01

    Full Text Available Neste trabalho, avaliou-se a capacidade fungitóxica do óleo essencial de folhas frescas de Tanaecium nocturnum sobre o Aspergillus flavus isolado da castanha-do-brasil, por meio das técnicas de contato e fumigação. Pelos resultados dos bioensaios realizados até 10 dias de incubação, verificou-se que a inibição total do crescimento micelial ocorreu quando se utilizou o óleo essencial nas concentrações de 782 ppm (técnica de contato e 1000 ppm (técnica de fumigação. Em ambas as técnicas, o óleo essencial inibiu a esporulação a partir da concentração de 500 ppm. Observou-se que nos cinco primeiros dias de incubação não houve diferença significativa nos resultados apresentados pelas duas técnicas estudadas, havendo a partir daí uma redução da atividade do óleo essencial nas concentrações inferiores a 1000 ppm pelo teste de fumigação. A ação fungitóxica do óleo essencial sobre o microrganismo estudado pode ser atribuída à presença do benzaldeído (composto majoritário do óleo essencial estudado, em associação com outros compostos também presentes nesse óleo essencial, tais como; álcool benzílico, benzoato de benzila e mandelonitrila.The present work sought to evaluate the fungitoxic activity of the essential oil from fresh Tanaecium nocturnum fresh leaves on Aspergillus flavus isolated from Brazil nuts, using contact and fumigation techniques. The results of bioassays performed up to 10 days of incubation demonstrated that total inhibition of mycelial growth occurred when using the essential oil at concentrations of 782 ppm (contact technique and 1000 ppm (fumigation technique. In both techniques, the essential oil inhibited the formation of spores at the concentration of 500 ppm. No significant difference in the results presented by the two techniques was observed in the first five days of incubation. After this period, the essential oil showed a reduction in activity at concentrations lower than

  3. Microsatellite Typing To Trace Aspergillus flavus Infections in a Hematology Unit ▿

    Science.gov (United States)

    Hadrich, Inès; Makni, Fattouma; Ayadi, Ali; Ranque, Stéphane

    2010-01-01

    Assessing the relatedness of strains isolated from patients and their environment is instrumental in documenting the source of preventable health care-associated life-threatening Aspergillus flavus human infection clusters. The present study aimed at identifying and selecting suitable microsatellite markers for A. flavus typing. This typing scheme was then applied to investigate the A. flavus epidemiology within a hematology unit in Sfax, Tunisia. Use of a combination of five markers made it possible to discern clusters of isolates and to substantiate the genetic diversity of A. flavus within clusters. Isolates from Tunisia and Marseille, France, displayed distinct haplotypes, indicating a highly significant geographical structuring of A. flavus. The typing of clinical and environmental A. flavus isolates in a hematology unit provided insights into its hospital epidemiology. From a heterogeneous genetic background, a cluster indicative of a clonal propagation episode within the unit could be identified. In two patients with invasive aspergillosis, the same genotype was found in clinical and environmental isolates, indicating hospital-acquired colonization and infection. In further studies, this novel microsatellite typing scheme might be instrumental in illuminating important epidemiological issues about A. flavus population genetics or epidemiology, including tracing the sources and routes of transmission. PMID:20410353

  4. Molecular characteristics of penicillin-binding protein 2b, 2x and 1a sequences in Streptococcus pneumoniae isolates causing invasive diseases among children in Northeast China.

    Science.gov (United States)

    Zhou, X; Liu, J; Zhang, Z; Liu, Y; Wang, Y; Liu, Y

    2016-04-01

    Streptococcus pneumoniae is one of the common pathogens causing severe invasive infections in children. This study aimed to investigate the serotype distribution and variations of penicillin-binding proteins (PBPs) 2b, 2x and 1a in S. pneumoniae isolates causing invasive diseases in Northeast China. A total of 256 strains were isolated from children with invasive pneumococcal disease (IPD) from January 2000 to October 2014. All strains were serotyped and determined for antibiotic resistance. The amplicons of penicillin-binding domains in pbp1a, pbp2b and pbp2x genes were sequenced for variation identification. The most prevalent serotypes of isolates in IPD children were 19A, 14, 19F, 23F and 6B. 19A and 19F were the most frequent serotypes of penicillin-resistant S. pneumoniae (PRSP), which present with high resistance to amoxicillin, cefotaxime, ceftriaxone and meropenem. The numbers of amino acid substitutions of penicillin-non-susceptible S. pneumoniae (PNSP) isolates were higher than those of penicillin-sensitive S. pneumoniae isolates in all the PBP genes (p isolates carried 25 amino acid mutations, including Ala618 → Gly between positions 560 and 675 in PBP2b and Thr338 → Ala substitutions in PBP2x. The amino acid alterations in PBP2b, PBP2x and PBP1a from S. pneumoniae were closely associated with resistance to β-lactam antibiotics. This study provides new data for further monitoring of genetic changes related to the emergence and spread of resistance to β-lactam antibiotics in China.

  5. Suppression of Aflatoxin Production in Aspergillus Species by Selected Peanut (Arachis hypogaea) Stilbenoids.

    Science.gov (United States)

    Sobolev, Victor; Arias, Renee; Goodman, Kerestin; Walk, Travis; Orner, Valerie; Faustinelli, Paola; Massa, Alicia

    2018-01-10

    Aspergillus flavus is a soil fungus that commonly invades peanut seeds and often produces carcinogenic aflatoxins. Under favorable conditions, the fungus-challenged peanut plant produces and accumulates resveratrol and its prenylated derivatives in response to such an invasion. These prenylated stilbenoids are considered peanut antifungal phytoalexins. However, the mechanism of peanut-fungus interaction has not been sufficiently studied. We used pure peanut stilbenoids arachidin-1, arachidin-3, and chiricanine A to study their effects on the viability of and metabolite production by several important toxigenic Aspergillus species. Significant reduction or virtually complete suppression of aflatoxin production was revealed in feeding experiments in A. flavus, Aspergillus parasiticus, and Aspergillus nomius. Changes in morphology, spore germination, and growth rate were observed in A. flavus exposed to the selected peanut stilbenoids. Elucidation of the mechanism of aflatoxin suppression by peanut stilbenoids could provide strategies for preventing plant invasion by the fungi that produce aflatoxins.

  6. Serotype changes and antimicrobial nonsusceptibility rates of invasive and non-invasive Streptococcus pneumoniae isolates after implementation of 10-valent pneumococcal nontypeable Haemophilus influenzae protein D conjugate vaccine (PHiD-CV) in Bulgaria.

    Science.gov (United States)

    Setchanova, Lena; Murdjeva, Marianna; Stancheva, Iglika; Alexandrova, Alexandra; Sredkova, Maria; Stoeva, Temenuga; Yoneva, Magda; Kurchatova, Anna; Mitov, Ivan

    The 10-valent pneumococcal conjugate vaccine (PCV10) has been included in Bulgarian Childhood Immunization Program since 2010. This study aimed to assess serotype distribution and antimicrobial resistance of 198 invasive and non-invasive Streptococcus pneumoniae strains that had been isolated in Bulgaria during 2011-2016 from patients with invasive (IPD) and non-invasive (NIPD) pneumococcal diseases. The most common invasive serotypes were 3 (10.1%), 19F (4.0%), and 7F (3.0%). A significant decrease in the proportion of invasive vaccine types (VTs) from 64.2% to 35.2% was found in comparison with pre-vaccine era. The most common serotypes among middle ear fluids were 3, 19A and 19F (5.6% each), and VTs fell down from 66.4% to 40.0% in post-PCV10 period. Among respiratory isolates, the most prevalent serotypes were some emergent serotypes such as 15A/B/C (5.0%), 19A, and 6C (4.0% each). VTs decreased significantly (16.3%) among vaccinated children compared to unvaccinated children and adults (44.0%). Two non-VTs (19A and 6C) have increased significantly more (pantibiotic nonsusceptible S. pneumoniae in Bulgaria remained high in post-PCV10 era. Among all source of isolates, antimicrobial nonsusceptibility rates were: oral penicillin - 46.5%, trimethoprim-sulfamethoxazole - 45.4%, erythromycin - 43.9%, tetracycline - 37.4%, and multidrug-resistance (MDR) was 44%. The most common MDR serotypes were 19F, 19A, 6A/C, 15A/B/C and 23A. Our results proved that PCV10 vaccination substantially reduced VTs pneumococcal IPD and NIPD. There has been a shift in the distribution of S. pneumoniae serotypes mostly in vaccinated children but also in the whole population and strong serotype-specific antibiotic resistance was observed after vaccine implementation. Therefore, it is important to continue monitoring serotype changes and pneumococcal resistance among all patient ages in addition to aid in determining the long-term effectiveness of PCV10 interventions. Copyright © 2017

  7. Serotype changes and antimicrobial nonsusceptibility rates of invasive and non-invasive Streptococcus pneumoniae isolates after implementation of 10-valent pneumococcal nontypeable Haemophilus influenzae protein D conjugate vaccine (PHiD-CV in Bulgaria

    Directory of Open Access Journals (Sweden)

    Lena Setchanova

    2017-07-01

    Full Text Available The 10-valent pneumococcal conjugate vaccine (PCV10 has been included in Bulgarian Childhood Immunization Program since 2010. This study aimed to assess serotype distribution and antimicrobial resistance of 198 invasive and non-invasive Streptococcus pneumoniae strains that had been isolated in Bulgaria during 2011–2016 from patients with invasive (IPD and non-invasive (NIPD pneumococcal diseases. The most common invasive serotypes were 3 (10.1%, 19F (4.0%, and 7F (3.0%. A significant decrease in the proportion of invasive vaccine types (VTs from 64.2% to 35.2% was found in comparison with pre-vaccine era. The most common serotypes among middle ear fluids were 3, 19A and 19F (5.6% each, and VTs fell down from 66.4% to 40.0% in post-PCV10 period. Among respiratory isolates, the most prevalent serotypes were some emergent serotypes such as 15A/B/C (5.0%, 19A, and 6C (4.0% each. VTs decreased significantly (16.3% among vaccinated children compared to unvaccinated children and adults (44.0%. Two non-VTs (19A and 6C have increased significantly more (p < 0.05 in vaccinated children than in unvaccinated patients. The rates of antibiotic nonsusceptible S. pneumoniae in Bulgaria remained high in post-PCV10 era. Among all source of isolates, antimicrobial nonsusceptibility rates were: oral penicillin – 46.5%, trimethoprim-sulfamethoxazole – 45.4%, erythromycin – 43.9%, tetracycline – 37.4%, and multidrug-resistance (MDR was 44%. The most common MDR serotypes were 19F, 19A, 6A/C, 15A/B/C and 23A. Our results proved that PCV10 vaccination substantially reduced VTs pneumococcal IPD and NIPD. There has been a shift in the distribution of S. pneumoniae serotypes mostly in vaccinated children but also in the whole population and strong serotype-specific antibiotic resistance was observed after vaccine implementation. Therefore, it is important to continue monitoring serotype changes and pneumococcal resistance among all patient ages in addition to

  8. The volatome of Aspergillus fumigatus.

    Science.gov (United States)

    Heddergott, C; Calvo, A M; Latgé, J P

    2014-08-01

    Early detection of invasive aspergillosis is absolutely required for efficient therapy of this fungal infection. The identification of fungal volatiles in patient breath can be an alternative for the detection of Aspergillus fumigatus that still remains problematic. In this work, we investigated the production of volatile organic compounds (VOCs) by A. fumigatus in vitro, and we show that volatile production depends on the nutritional environment. A. fumigatus produces a multiplicity of VOCs, predominantly terpenes and related compounds. The production of sesquiterpenoid compounds was found to be strongly induced by increased iron concentrations and certain drugs, i.e., pravastatin. Terpenes that were always detectable in large amounts were α-pinene, camphene, and limonene, as well as sesquiterpenes, identified as α-bergamotene and β-trans-bergamotene. Other substance classes that were found to be present in the volatome, such as 1-octen-3-ol, 3-octanone, and pyrazines, were found only under specific growth conditions. Drugs that interfere with the terpene biosynthesis pathway influenced the composition of the fungal volatome, and most notably, a block of sesquiterpene biosynthesis by the bisphosphonate alendronate fundamentally changed the VOC composition. Using deletion mutants, we also show that a terpene cyclase and a putative kaurene synthase are essential for the synthesis of volatile terpenes by A. fumigatus. The present analysis of in vitro volatile production by A. fumigatus suggests that VOCs may be used in the diagnosis of infections caused by this fungus. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  9. The impact of the increased use of piperacillin/tazobactam on the selection of antibiotic resistance among invasive Escherichia coli and Klebsiella pneumoniae isolates.

    Science.gov (United States)

    Lee, Jina; Oh, Chi Eun; Choi, Eun Hwa; Lee, Hoan Jong

    2013-08-01

    Increasing antimicrobial resistance is related to the selective pressure exerted by antibiotic usage. This study evaluated the impact of the increased use of piperacillin/tazobactam (TZP) on the selection of antibiotic resistance. From 1999 to 2010, Escherichia coli and Klebsiella pneumoniae invasive isolates obtained from hospitalized Korean children were analyzed in antibiotic susceptibility tests and subjected to characterization of the β-lactamase types. Antibiotic consumption data were also analyzed. Between January 1999 and December 2010, 409 invasive isolates of E. coli (n=170) and K. pneumoniae (n=239) were obtained. A rebound of extended-spectrum β-lactamase (ESBL) prevalence with an increase in total antibiotics was noted. Non-susceptibility to TZP was determined in 7.6% of E. coli isolates and 20.9% of K. pneumoniae isolates. Despite the increase in TZP usage, the overall prevalence of TZP resistance did not significantly increase over time, especially in E. coli. The mechanisms for TZP resistance included the presence of AmpC producers, possible TEM-1 hyperproducers, and multiple β-lactamases in individual organisms of a given isolate. Replacement of only the antibiotic class appears to be insufficient to control antibiotic resistance, and continued efforts to decrease overall antibiotic pressure are needed, especially in highly endemic situations. Copyright © 2013 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  10. Chemodiversity in the genus Aspergillus

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Larsen, Thomas Ostenfeld

    2015-01-01

    to be characterized. The genus Aspergillus is cladistically holophyletic but phenotypically polythetic and very diverse and is associated to quite different sexual states. Following the one fungus one name system, the genus Aspergillus is restricted to a holophyletic clade that include the morphologically different...... biosynthetic family isoextrolites. However, it appears that secondary metabolites from one Aspergillus section have analogous metabolites in other sections (here also called heteroisoextrolites). In this review, we give a genus-wide overview of secondary metabolite production in Aspergillus species. Extrolites...

  11. Comparative studies on pectinases obtained from Aspergillus ...

    African Journals Online (AJOL)

    Comparative studies on pectinases obtained from Aspergillus fumigatus and Aspergillus niger in submerged fermentation system using pectin extracted from mango, orange and pineapple peels as carbon sources.

  12. Invasive cranial mycosis our experiences

    Directory of Open Access Journals (Sweden)

    Tapas Kumbhkar

    2013-01-01

    Full Text Available Fungi can cause serious cranial infections in immunocompromised and diabetic patients. Common pathogens mainly include Aspergillus and Mucor. These organisms cause tissue invasion and destruction of adjacent structures (e.g. orbit, ethmoid, sphenoid, maxillary & cavernous sinuses. Mortality and morbidity rate is high despite combined surgical, antifungal and antidiabetic treatment. We present our experience of six cases with such infection.

  13. Population dynamics of Aspergillus section Nigri species on vineyard samples of grapes and raisins

    Science.gov (United States)

    Several species of Aspergillus section Nigri, including potential mycotoxin producers, are common residents of grape vineyards, but the relative population size of individual species throughout the growing season is difficult to determine using traditional isolation and identification methods. Using...

  14. Streptococcus pneumoniae aislados de infecciones invasivas: serotipos y resistencia antimicrobiana Streptococcus pneumoniae isolated from invasive infections: serotypes and antimicrobial resistance

    Directory of Open Access Journals (Sweden)

    Gladys Antonia Cueto Montoya

    2007-03-01

    Full Text Available Las meningoencefalitis bacterianas constituyen una enfermedad invasiva importante, quizás no tanto por su frecuencia, como por la gravedad de su cuadro. Los cambios en la epidemiología de los síndromes neurológicos infecciosos en Cuba a partir de la vacunación contra meningococo BC y Haemophilus influenzae b han hecho que el Streptococcus pneumoniae constituya el agente causal más frecuente. Debido al incremento de la resistencia de este microorganismo a los antibióticos habituales, se realizaron modificaciones al régimen terapéutico convencional, fundamentalmente en las meningitis pediátricas. Es necesario lograr el aislamiento en cultivo de este agente para conocer los serotipos más frecuentes en el país, y lograr una vacuna neumocócica conjugada, así como para la vigilancia de las cepas frente a los antimicrobianos.The bacterial meningoencephalitis is an important invasive disease, not only because of its frequency, but also because of the severity of its picture. The changes in the epidemiology of the neurological infectious syndromes in Cuba starting from the vaccination against meningococcus BC and Haemophilus infuenzae b have made that Streptococcus pneumoniae be the most frequent causal agent. Due to the increase of the resistance of this microorganism to habitual antibiotics, modifications were made in the conventional therapeutic regimen, mainly in the pediatric meningitis. It is necessary to achieve the isolation in culture of this agent to know the most common serotypes in the country, to attain a conjugated pneumococcal vaccine, and to keep the surveillance of the strains against the antimicrobials.

  15. Genomic comparison of multi-drug resistant invasive and colonizing Acinetobacter baumannii isolated from diverse human body sites reveals genomic plasticity

    Directory of Open Access Journals (Sweden)

    Hsiao William W

    2011-06-01

    Full Text Available Abstract Background Acinetobacter baumannii has recently emerged as a significant global pathogen, with a surprisingly rapid acquisition of antibiotic resistance and spread within hospitals and health care institutions. This study examines the genomic content of three A. baumannii strains isolated from distinct body sites. Isolates from blood, peri-anal, and wound sources were examined in an attempt to identify genetic features that could be correlated to each isolation source. Results Pulsed-field gel electrophoresis, multi-locus sequence typing and antibiotic resistance profiles demonstrated genotypic and phenotypic variation. Each isolate was sequenced to high-quality draft status, which allowed for comparative genomic analyses with existing A. baumannii genomes. A high resolution, whole genome alignment method detailed the phylogenetic relationships of sequenced A. baumannii and found no correlation between phylogeny and body site of isolation. This method identified genomic regions unique to both those isolates found on the surface of the skin or in wounds, termed colonization isolates, and those identified from body fluids, termed invasive isolates; these regions may play a role in the pathogenesis and spread of this important pathogen. A PCR-based screen of 74 A. baumanii isolates demonstrated that these unique genes are not exclusive to either phenotype or isolation source; however, a conserved genomic region exclusive to all sequenced A. baumannii was identified and verified. Conclusions The results of the comparative genome analysis and PCR assay show that A. baumannii is a diverse and genomically variable pathogen that appears to have the potential to cause a range of human disease regardless of the isolation source.

  16. Biodiversity of species of Aspergillus section Fumigati in semi-desert soils in Argentina.

    Science.gov (United States)

    Giusiano, Gustavo E; Piontelli, Eduardo; Fernández, Mariana S; Mangiaterra, Magdalena L; Cattana, María E; Kocsubé, Sándor; Varga, János

    The distribution of Aspergillus species in soil has been widely studied all over the world. The aim of this study was the phenotypic and genotypic characterization of species Aspergillus belonging to section Fumigati present in soils from two Argentinian semi-desert areas having different geological conditions. Altogether, 23 isolates belonging to Aspergillus section Fumigati were recovered and identified using a polyphasic approach including phenotypic and molecular identifications. Aspergillus fumigatus sensu stricto and Aspergillus fumigatiaffinis had the highest frequency, of occurrence while isolates closely related to Aspergillus udagawae and Aspergillus felis were rarely observed. A. fumigatiaffinis and isolates closer to A. udagawae were isolated for the first time from Argentinian soils and this is the first report on the occurrence of species belonging to the A. felis clade in South America. Recent scientific interests in biodiversity, as well as the increasing importance of aspergilli as causative agents of human and animal diseases increase the need to understand the diversity and occurrence of these fungi in nature. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  17. Detection of Aflatoxin Producing Aspergillus flavus in Post-harvest Contaminated Vigna ungulculata Seeds

    OpenAIRE

    Ajay Kumar Gautam

    2014-01-01

    The present study was carried out with a specific objective to study postharvest spoilage of Lobhiya (Vigna unguiculata) seeds contaminated with Aspergillus flavus. Infected seeds were collected and cultured on potato dextrose agar (PDA) media, at 25±2 °C. Aspergillus flavus isolates were primarily characterized by its morphological and microscopic characteristics. Collected fungal isolates were also screened for their afaltoxigenic nature on preliminary basis and at molecul...

  18. Aspergillus section Versicolores: nine new species and multilocus DNA sequence based phylogeny

    OpenAIRE

    Jurjevic, Zeljko; Peterson, Stephen W.; Horn, Bruce W.

    2012-01-01

    β-tubulin, calmodulin, internal transcribed spacer and partial lsu-rDNA, RNA polymerase 2, DNA replication licensing factor Mcm7, and pre-rRNA processing protein Tsr1 were amplified and sequenced from numerous isolates belonging to Aspergillus sect. versicolor. The isolates were analyzed phylogenetically using the concordance model to establish species boundaries. Aspergillus austroafricanus, A. creber, A. cvjetkovicii, A. fructus, A. jensenii, A. puulaauensis, A. subversicolor, A. tennesseen...

  19. Aspergillus alabamensis, a New Clinically Relevant Species in the Section Terrei

    DEFF Research Database (Denmark)

    Balajee, S. A.; Baddley, J. W.; Peterson, S. W.

    2009-01-01

    Phylogenetic analyses of sequences generated from portions of three genes coding for the proteins enolase (enoA), beta-tubulin (benA), and calmodulin (calM) of a large number of isolates within the section Terrei, genus Aspergillus, revealed the presence of a new cryptic species within this section......, Aspergillus alabamensis. Most members of this new cryptic species were recovered as colonizing isolates from immunocompetent patient populations, had decreased in vitro susceptibilities to the antifungal drug amphotericin B, and were morphologically similar to but genetically distinct from Aspergillus terreus...

  20. Cellulase production by wild strains of Aspergillus niger, Penicillium ...

    African Journals Online (AJOL)

    Waste cellulosic materials (corncob, sawdust and sugarcane pulp) and crystalline cellulose induced cellulase production in wild strains of Aspergillus niger, Penicillium chrysogenum and Trichoderma harzianum isolated from a wood-waste dump in Lagos, Nigeria. Cellulose-supplemented media gave the maximum ...

  1. Method for measuring postantifungal effect in Aspergillus species.

    NARCIS (Netherlands)

    Vitale, R.G.; Mouton, J.W.; Afeltra, J.; Meis, J.F.G.M.; Verweij, P.E.

    2002-01-01

    An in vitro method for determination of postantifungal effect (PAFE) in molds was developed by using three isolates each of Aspergillus fumigatus, A. flavus, A. terreus, A. nidulans, and A. ustus. MICs of amphotericin B and itraconazole were determined by using National Committee for Clinical

  2. Production of emodin from Aspergillus ochraceus at preparative scale

    African Journals Online (AJOL)

    ONOS

    2010-01-25

    Jan 25, 2010 ... In order to study the chemical constituents in the pigmented culture produced from Aspergillus ochraceus, solid phase extraction method was employed to isolate the pigment molecules from the primary culture, followed by fractionation on preparative liquid chromatography. Structural characterization ...

  3. Full Length Research Paper Production of emodin from Aspergillus ...

    African Journals Online (AJOL)

    In order to study the chemical constituents in the pigmented culture produced from Aspergillus ochraceus, solid phase extraction method was employed to isolate the pigment molecules from the primary culture, followed by fractionation on preparative liquid chromatography. Structural characterization confirmed that one of ...

  4. An amendment of Aspergillus section Candidi based on chemotaxonomical evidence

    DEFF Research Database (Denmark)

    Rahbaek, L.; Frisvad, Jens Christian; Christophersen, C.

    2000-01-01

    A novel 2,2 '-epoxy-terphenyllin, candidusin C, in addition to the well known secondary metabolites terphenyllin, 3-hydroxyterpenyllin and chlorflavonin, has been isolated from the chemically unexplored fungus Aspergillus campestris. The latter three are known secondary metabolites from Aspergill...

  5. Production and partial purification of glucoamylase from Aspergillus ...

    African Journals Online (AJOL)

    SAM

    2014-05-21

    May 21, 2014 ... Glucoamylase is an enzyme that hydrolyses 1,4α and 1,6β-glucosidic linkages in polysaccharides yielding glucose. Aspergillus niger strains 1, 2 and 3 were locally isolated from cassava peel dumpsite for the production of glucoamylase enzyme. A. niger strains 1, 2 and 3 were screened for their hyper.

  6. Production and partial purification of glucoamylase from Aspergillus ...

    African Journals Online (AJOL)

    Glucoamylase is an enzyme that hydrolyses 1,4α and 1,6β-glucosidic linkages in polysaccharides yielding glucose. Aspergillus niger strains 1, 2 and 3 were locally isolated from cassava peel dumpsite for the production of glucoamylase enzyme. A. niger strains 1, 2 and 3 were screened for their hyper producing ability on ...

  7. Partially purified polygalacturonase from Aspergillus niger (SA6 ...

    African Journals Online (AJOL)

    Polygalacturonase (PG) was isolated from Aspergillus niger (A. niger) (SA6), partially purified and characterized. The PG showed two bands on SDS-PAGE suggesting an “endo and exo PG with apparent molecular weights of 35 and 40 KDa, respectively. It was purified 9-fold with a yield of 0.18% and specific activity of 246 ...

  8. The occurrence of a phosphorylated glycosphingolipid in Aspergillus niger.

    Science.gov (United States)

    Brennan, P J; Roe, J

    1975-01-01

    A novel type of water-soluble phosphorylated glycosphingolipid was isolated from Aspergillus niger by a simple procedure involving precipitation, DEAE-cellulose chromatography and preparative t.l.c. Besides ceramide and phosphorus it contains inositol, galactose, mannose and small amounts of glucosamine. Images PLATE 1 PMID:1156383

  9. Performance of Molecular Approaches for Aspergillus Detection and Azole Resistance Surveillance in Cystic Fibrosis

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    Hélène Guegan

    2018-03-01

    Full Text Available Aspergillus fumigatus triazole resistance is an emerging concern for treating chronically infected/colonized patients. This study sought to evaluate the performance of PCR assays to detect Aspergillus fungi together with azole resistance in sputum samples from cystic fibrosis (CF patients. In total, 119 sputum samples from 87 CF patients were prospectively processed for Aspergillus detection by means of mycological culture and four qPCR assays, 2 in-house methods and two commercial multiplex real-time PCR assays simultaneously detecting Aspergillus and the most relevant cyp51A gene mutations (MycoGENIE® and AsperGenius®. Azole susceptibility of A. fumigatus isolates was assessed using Etest® method and cyp51A gene mutation were characterized by sequencing. The overall rate of Aspergillus detection with the four qPCR assays ranged from 47.9 to 57.1%, contrasting with 42/119 (35.3% positive cultures with A. fumigatus. The high sensitivity of PCR on sputum could then contribute to more effective grading of Aspergillus disease in CF patients. Five out of 41 isolated strains (12.2% exhibited azole-resistant MIC patterns, three of which harbored cyp51A mutations and only 1/3 with the sequence TR34/L98H. Combined with culture, PCR assay achieved high sensitivity Aspergillus screening in CF samples. However, cyp51A targeting was only moderately effective for azole resistance monitoring, while Aspergillus resistance remains of great concern.

  10. Pattern recognition receptors and their role in invasive aspergillosis

    NARCIS (Netherlands)

    Gresnigt, M.S.; Netea, M.G.; van de Veerdonk, F.L.

    2012-01-01

    Pattern recognition receptors (PRRs) are germline receptors that recognize conserved structures on microorganisms. Several PRRs have been identified in the recent years that are involved in the immune response against Aspergillus fumigatus. The role of PRRs in invasive pulmonary aspergillosis

  11. Characterization of Salmonella Gallinarum isolates from backyard poultry by polymerase chain reaction detection of invasion (invA and Salmonella plasmid virulence (spvC genes

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    Susmita Pal

    2017-07-01

    Full Text Available Aim: The aim was to characterize Salmonella enterica serovar Gallinarum isolated from backyard poultry by polymerase chain reaction (PCR detection of virulence genes invasion (invA and Salmonella plasmid virulence C (spvC. Materials and Methods: Two strains of Salmonella serovar Gallinarum isolates used in this study were obtained from an outbreak of fowl typhoid in backyard Vanaraja fowl. PCR technique was used for detection of invA and spvC genes using standard methodology. The invA PCR product from one representative isolate was sequenced and compared with other related Salmonella serovars in GenBank data. Results: Salmonella Gallinarum produced expected amplicons of invA and spvC gene products. Nucleotide sequence of 285 bp invA gene was deposited in GenBank with accession no. KX788214. Sequence analysis of invA gene was found conserved in Salmonella serovars and demonstrated 100% homology with closely related serovars of Salmonella. Conclusion: Invasion gene (invA was found to be highly conserved in Salmonella Gallinarum and highly similar with closely related serovars. The isolates also contained plasmid-mediated spvC gene indicating possession of virulence plasmid.

  12. Genetic characterisation of virulence genes associated with adherence, invasion and cytotoxicity in Campylobacter spp. isolated from commercial chickens and human clinical cases

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    Samantha Reddy

    2018-02-01

    Full Text Available Virulence-associated genes have been recognised and detected in Campylobacter species. The majority of them have been proven to be associated with pathogenicity. This study aimed to detect the presence of virulence genes associated with pathogenicity and responsible for invasion, expression of adherence, colonisation and production of the cytolethal distending toxin (cdt in Campylobacter jejuni and Campylobacter coli. Commercial chicken faecal samples were randomly sampled from chicken farms within the Durban metropolitan area in South Africa. Furthermore, human clinical Campylobacter spp. isolates were randomly sampled from a private pathology laboratory in South Africa. Out of a total of 100 chicken faecal samples, 78% (n = 78 were positive for Campylobacter growth on modified charcoal cefoperazone deoxycholate and from the random laboratory collection of 100 human clinical isolates, 83% (n = 83 demonstrated positive Campylobacter spp. growth following culturing methods. These samples were screened for the presence of the following virulence genes: cadF, hipO, asp, ciaB, dnaJ, pldA, cdtA, cdtB and cdtC. As expected, the cadF gene was present in 100% of poultry (n = 78 and human clinical isolates (n = 83. Campylobacter jejuni was the main species detected in both poultry and human clinical isolates, whilst C. coli were detected at a significantly lower percentage (p < 0.05. Eight per cent of the C. jejuni from human clinical isolates had all virulence genes that were investigated. Only one C. coli isolate demonstrated the presence of all the virulence genes investigated; however, the pldA virulence gene was detected in 100% of the C. coli isolates in poultry and a high percentage (71% in human clinical C. coli isolates as well. The detection of cdt genes was found at higher frequency in poultry than human clinical isolates. The high prevalence rates of virulence genes detected in poultry and human clinical isolates demonstrate their

  13. Aspergillus Sydowi Infection of Human Finger Nail

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    A K Barde

    1981-01-01

    Full Text Available A case of Aspergillus sydowi infection of left middle finger nail is described ′ The presence of fungal hypae with phialids and spores on direct microscopy as well as in culture, the colour of the sub-ungual mass of the nail resembling the colour of the fungus in, culture′ repeated isolations of A sydowi from the diseased tissue along with the absence of any established pathogenic species in the specimen are taken as evidences that this fungus was invading the nail tissue.

  14. Aspergillus bertholletius sp. nov. from Brazil Nuts

    DEFF Research Database (Denmark)

    Taniwaki, Marta H.; Pitt, John I.; Iamanaka, Beatriz T.

    2012-01-01

    During a study on the mycobiota of brazil nuts (Bertholletia excelsa) in Brazil, a new Aspergillus species, A. bertholletius, was found, and is described here. A polyphasic approach was applied using morphological characters, extrolite data as well as partial beta-tubulin, calmodulin and ITS...... sequences to characterize this taxon. A. bertholletius is represented by nineteen isolates from samples of brazil nuts at various stages of production and soil close to Bertholletia excelsa trees. The following extrolites were produced by this species: aflavinin, cyclopiazonic acid, kojic acid, tenuazonic...

  15. New and revisited species in Aspergillus section Nigri

    DEFF Research Database (Denmark)

    Varga, J.; Frisvad, Jens Christian; Kocsube, S.

    2011-01-01

    Four new species, Aspergillus eucalypticola, A. neoniger, A. fijiensis and A. indolo genus are described and illustrated. Aspergillus eucalypticola was isolated from Eucalyptus leaf from Australia, and is related to A. tubingensis and A. costaricaensis, but could clearly be distinguished from them...... are also detailed. Although these species differ in their occurrence and several physiological means (elastase activities, abilities to utilise 2-deoxy-D-glucose as sole carbon source), our data indicate that only molecular approaches including sequence analysis of calmodulin or beta-tubulin genes, AFLP...

  16. Antimicrobial resistance among isolates causing invasive pneumococcal disease before and after licensure of heptavalent conjugate pneumococcal vaccine.

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    Tom Theodore Karnezis

    Full Text Available BACKGROUND: The impact of the pneumococcal conjugate vaccine (PCV-7 on antibiotic resistance among pneumococcal strains causing invasive pneumococcal disease (IPD has varied in different locales in the United States. We assessed trends in IPD including trends for IPD caused by penicillin non-susceptible strains before and after licensure of PCV-7 and the impact of the 2008 susceptibility breakpoints for penicillin on the epidemiology of resistance. METHODOLOGY/PRINCIPAL FINDINGS: We performed a retrospective review of IPD cases at Morgan Stanley Children's Hospital of New York-Presbyterian, Columbia University Medical Center. Subjects were < or = 18 years of age with Streptococcus pneumoniae isolated from sterile body sites from January 1995-December 2006. The rate of IPD from 1995-1999 versus 2002-2006 significantly decreased from 4.1 (CI(95 3.4, 4.8 to 1.7 (CI(95 1.3, 2.2 per 1,000 admissions. Using the breakpoints in place during the study period, the proportion of penicillin non-susceptible strains increased from 27% to 49% in the pre- vs. post-PCV-7 era, respectively (p = 0.001, although the rate of IPD caused by non-susceptible strains did not change from 1995-1999 (1.1 per 1,000 admissions, CI(95 0.8, 1.5 when compared with 2002-2006 (0.8 per 1,000 admissions, CI(95 0.6, 1.2. In the multivariate logistic regression model controlling for the effects of age, strains causing IPD in the post-PCV-7 era were significantly more likely to be penicillin non-susceptible compared with strains in the pre-PCV-7 era (OR 2.46, CI(95 1.37, 4.40. However, using the 2008 breakpoints for penicillin, only 8% of strains were non-susceptible in the post-PCV-7 era. CONCLUSIONS/SIGNIFICANCE: To date, there are few reports that document an increase in the relative proportion of penicillin non-susceptible strains of pneumococci causing IPD following the introduction of PCV-7. Active surveillance of pneumococcal serotypes and antibiotic resistance using the new

  17. Use of Epidemiological Cutoff Values To Examine 9-Year Trends in Susceptibility of Aspergillus Species to the Triazoles▿

    Science.gov (United States)

    Pfaller, M.; Boyken, L.; Hollis, R.; Kroeger, J.; Messer, S.; Tendolkar, S.; Diekema, D.

    2011-01-01

    In the absence of clinical breakpoints, epidemiological cutoff values (ECVs) have been established to distinguish wild-type (WT) isolates of Aspergillus spp. from those that may harbor resistance mutations. Recently, the CLSI has developed ECVs for triazoles (itraconazole, posaconazole, and voriconazole) and common Aspergillus species. We applied the triazole ECVs to 1,789 Aspergillus isolates collected from 63 centers worldwide from 2001 to 2009 to determine the frequency of non-WT strains of each species. Temporal trends were evaluated for Aspergillus fumigatus and Aspergillus flavus over the 9-year period for each drug. The collection included 1,312 isolates of A. fumigatus, 235 of A. flavus, 162 of Aspergillus niger, 64 of Aspergillus terreus, and 15 of Aspergillus versicolor. Using the ECVs, the percentages of non-WT isolates for itraconazole, posaconazole, and voriconazole, respectively, were as follows: A. fumigatus (2.0%, 3.5%, and 1.4%), A. flavus (0.8%, 5.1%, and 1.7%), A. niger (17.3%, 3.7%, and 0.6%), A. terreus (0.0%, 1.6%, and 3.2%), and A. versicolor (6.3%, 0.0%, and 0.0%). Among 49 Aspergillus isolates for which itraconazole MICs were >2 μg/ml, the posaconazole and voriconazole MICs were greater than the ECVs for 14 and 12 isolates, respectively. The percentages of isolates for which MICs were greater than the ECVs ranged from 1.1 to 5.7% for posaconazole, 0.0 to 1.6% for voriconazole, and 0.7 to 4.0% for itraconazole. There was no consistent trend toward decreased susceptibility for any triazole and A. fumigatus or A. flavus over time. Decreased susceptibility among Aspergillus spp. was observed for each of the extended-spectrum triazoles and varied by species over the 9-year study period. PMID:21123534

  18. Polyphasic taxonomy of Aspergillus section Aspergillus (formerly Eurotium), and its occurrence in indoor environments and food

    DEFF Research Database (Denmark)

    Chen, A.J.; Hubka, Vit; Frisvad, Jens Christian

    2017-01-01

    Aspergillus section Aspergillus (formerly the genus Eurotium) includes xerophilic species with uniseriate conidiophores, globose to subglobose vesicles, green conidia and yellow, thin walled eurotium-like ascomata with hyaline, lenticular ascospores. In the present study, a polyphasic approach...... using morphological characters, extrolites, physiological characters and phylogeny was applied to investigate the taxonomy of this section. Over 500 strains from various culture collections and new isolates obtained from indoor environments and a wide range of substrates all over the world were......, epiheveadrides, auroglaucins and anthraquinone bisanthrons, and to be consistent in strains of nearly all species. Other extrolites are species-specific, and thus valuable for identification. Several extrolites show antioxidant effects, which may be nutritionally beneficial in food and beverages. Important...

  19. Genomic analysis reveals multi-drug resistance clusters in Group B Streptococcus CC17 hypervirulent isolates causing neonatal invasive disease in southern mainland China

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    Edmondo Campisi

    2016-08-01

    Full Text Available Neonatal invasive disease caused by group B Streptococcus (GBS represents a significant public health care concern globally. However, data related to disease burden, serotype distribution and molecular epidemiology in China and other Asian countries are very few and specifically relative to confined regions. The aim of this study was to investigate the genetic characteristics of GBS isolates recovered from neonates with invasive disease during 2013-2014 at Guangzhou and Changsha hospitals in southern mainland China. We assessed the capsular polysaccharide (CPS type, pilus islands (PIs distribution and hvgA gene presence in a panel of 26 neonatal clinical isolates, of which 8 were recovered from Early Onset Disease (EOD and 18 from Late Onset Disease (LOD. Among 26 isolates examined, five serotypes were identified. Type III was the most represented (15 cases, particularly among LOD strains (n=11, followed by types Ib (n=5, V (n=3, Ia (n=2 and II (n=1. We performed whole-genome sequencing (WGS analysis and antimicrobial susceptibility testing on the 14 serotype III isolates belonging to the hypervirulent Clonal Complex 17 (serotype III-CC17.The presence of PI-2b alone was associated with 13 out of 14 serotype III-CC17 strains. Genome analysis led us to identify two multi-drug resistance gene clusters harbored in two new versions of integrative and conjugative elements (ICEs, carrying five or eight antibiotic resistance genes, respectively. These ICEs replaced the 16 kb-locus that normally contains the PI-1 operon. All isolates harboring the identified ICEs showed multiple resistances to aminoglycoside, macrolide and tetracycline antibiotic classes. In conclusion, we report the first whole-genome sequence analysis of 14 GBS serotype III-CC17 strains isolated in China, representing the most prevalent lineage causing neonatal invasive disease. The acquisition of newly identified ICEs conferring multiple antibiotic resistances could in part explain

  20. Complement Attack against Aspergillus and Corresponding Evasion Mechanisms

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    Cornelia Speth

    2012-01-01

    Full Text Available Invasive aspergillosis shows a high mortality rate particularly in immunocompromised patients. Perpetually increasing numbers of affected patients highlight the importance of a clearer understanding of interactions between innate immunity and fungi. Innate immunity is considered to be the most significant host defence against invasive fungal infections. Complement represents a crucial part of this first line defence and comprises direct effects against invading pathogens as well as bridging functions to other parts of the immune network. However, despite the potency of complement to attack foreign pathogens, the prevalence of invasive fungal infections is increasing. Two possible reasons may explain that phenomenon: First, complement activation might be insufficient for an effective antifungal defence in risk patients (due to, e.g., low complement levels, poor recognition of fungal surface, or missing interplay with other immune elements in immunocompromised patients. On the other hand, fungi may have developed evasion strategies to avoid recognition and/or eradication by complement. In this review, we summarize the most important interactions between Aspergillus and the complement system. We describe the various ways of complement activation by Aspergillus and the antifungal effects of the system, and also show proven and probable mechanisms of Aspergillus for complement evasion.

  1. Phosphate solubilizing ability of two Arctic Aspergillus niger strains

    Directory of Open Access Journals (Sweden)

    Shiv Mohan Singh,

    2011-06-01

    Full Text Available Many filamentous fungi were isolated from the soils of Ny-Ålesund, Spitsbergen, Svalbard, and were screened in vitro for their phosphate solubilizing ability. Two strains of Aspergillus niger showed good tricalcium phosphate (TCP solubilizing ability in Pikovskaya's medium. The TCP solubilization index was calculated at varying levels of pH and temperatures. The ability of Aspergillus niger strain-1 to solubilize and release inorganic-P was 285 µg ml–1, while Aspergillus niger strain-2 solubilized 262 µg ml–1 from 0.5% TCP after seven days. This is the first report of TCP solubilization by Arctic strains that may serve as very good phosphate solubilizers in the form of biofertilizer.

  2. Analysis of expressed sequence tags (ESTs) from a normalized cDNA library and isolation of EST simple sequence repeats from the invasive cotton mealybug Phenacoccus solenopsis.

    Science.gov (United States)

    Li, Hui; Lang, Kun-Ling; Fu, Hai-Bin; Shen, Chang-Peng; Wan, Fang-Hao; Chu, Dong

    2015-12-01

    The cotton mealybug, Phenacoccus solenopsis Tinsley, is a serious and invasive pest. At present, genetic resources for studying P. solenopsis are limited, and this negatively affects genetic research on the organism and, consequently, translational work to improve management of this pest. In the present study, expressed sequence tags (ESTs) were analyzed from a normalized complementary DNA library of P. solenopsis. In addition, EST-derived microsatellite loci (also known as simple sequence repeats or SSRs) were isolated and characterized. A total of 1107 high-quality ESTs were acquired from the library. Clustering and assembly analysis resulted in 785 unigenes, which were classified functionally into 23 categories according to the Gene Ontology database. Seven EST-based SSR markers were developed in this study and are expected to be useful in characterizing how this invasive species was introduced, as well as providing insights into its genetic microevolution. © 2014 Institute of Zoology, Chinese Academy of Sciences.

  3. In vitro activity of disinfectants against Aspergillus spp

    Directory of Open Access Journals (Sweden)

    A.S. Mattei

    2013-01-01

    Full Text Available Fungi of the Aspergillus genus are widespread and contaminate the environment. Thousands of conidia are released from each phialide and dispersed in the air every day. These fungi are considered important mycose-causing agents in hospitals. Due to this, research to determine prevalent fungi from the Aspergillus genus in hospital environments, and an adequate disinfection program in these areas is are needed. This study evaluated the susceptibility of Aspergillus spp. isolated from a veterinary environment against four disinfectants. Successive dilutions of disinfectants (log2 were used according to CLSI M38-A2 microdilution technique adapted to chemical agents against 18 isolates of this genus. After 72 hours of incubation, the Minimum Inhibiting Concentration and Minimum Fungicidal Concentration capable of inhibiting 50% and 90% of the isolates were determined. Chlorexidine-cetrimine, benzalconium chloride and a chlorophenol derivative proved to be effective against all isolates with a lower MIC than that suggested by the manufacturer, except for the A. flavus strain. Sodium hypochlorite was ineffective against three A. fumigatus, three A. flavus and one A. niger isolate. These results demonstrated that all studied disinfectants were effective against environmental isolates, with the exception of sodium hypochlorite, which showed lower effectiveness.

  4. Aspergillus subgenus Polypaecilum from the built environment

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    J.B. Tanney

    2017-09-01

    Full Text Available Xerophilic fungi, especially Aspergillus species, are prevalent in the built environment. In this study, we employed a combined culture-independent (454-pyrosequencing and culture-dependent (dilution-to-extinction approach to investigate the mycobiota of indoor dust collected from 93 buildings in 12 countries worldwide. High and low water activity (aw media were used to capture mesophile and xerophile biodiversity, resulting in the isolation of approximately 9 000 strains. Among these, 340 strains representing seven putative species in Aspergillus subgenus Polypaecilum were isolated, mostly from lowered aw media, and tentatively identified based on colony morphology and internal transcribed spacer rDNA region (ITS barcodes. Further morphological study and phylogenetic analyses using sequences of ITS, β-tubulin (BenA, calmodulin (CaM, RNA polymerase II second largest subunit (RPB2, DNA topoisomerase 1 (TOP1, and a pre-mRNA processing protein homolog (TSR1 confirmed the isolation of seven species of subgenus Polypaecilum, including five novel species: A. baarnensis, A. keratitidis, A. kalimae sp. nov., A. noonimiae sp. nov., A. thailandensis sp. nov., A. waynelawii sp. nov., and A. whitfieldii sp. nov. Pyrosequencing detected six of the seven species isolated from house dust, as well as one additional species absent from the cultures isolated, and three clades representing potentially undescribed species. Species were typically found in house dust from subtropical and tropical climates, often in close proximity to the ocean or sea. The presence of subgenus Polypaecilum, a recently described clade of xerophilic/xerotolerant, halotolerant/halophilic, and potentially zoopathogenic species, within the built environment is noteworthy.

  5. Two novel ail-positive biotype 1A strains of Yersinia enterocolitica isolated in China with unequal adhesion and invasion properties.

    Science.gov (United States)

    Liang, Junrong; Bi, Zhenqiang; Shi, Guoxiang; Xiao, Yuchun; Qiu, Haiyan; Kou, Zengqiang; Hu, Bin; Jing, Huaiqi; Wang, Xin

    2014-10-01

    Yersinia enterocolitica is an enteric pathogen having six biotypes: 1A, 1B, 2, 3, 4, and 5. Different bioserotypes have been associated with varying pathogenicity, and the strains of biotype 1A lack the virulence-associated pYV-bearing genes and were once considered to be avirulent. However, there is growing epidemiological, clinical, and experimental evidence to suggest some biotype 1A isolates are virulent and can cause gastrointestinal disease. Here, we describe two biotype 1A strains discovered from 3807 isolates that carry the ail (attachment and invasion locus) gene. The two strains showed unique PFGE patterns compared to all other isolates in the Chinese Y. enterocolitica isolate PFGE database. Strain SDWL-003 isolated from a sheep shared ail sequence identical to A1 pattern, and the foxA (ferrioxamine receptor) sequence was identical to the pathogenic F5 pattern, besides, the PFGE patterns of SDWL-003 was also cluster to pathogenic branch; however it does not attach to or invade Hep-2 cells. The ail sequence of strain 2006RAT isolated from a Microtus fortis showed several mutations compared to other published genomes, and therefore formed an entirely new pathogenic pattern. Though it clustered to non-pathogenic block with foxA sequence polymorphism analysis or PFGE assay, the strain 2006RAT showed adhesion properties. The data here bring new insights into the molecular genetics of Y. enterocolitica biotype 1A, show some isolates of 1A biotype gaining potential pathogenicity using the function of the virulence gene - ail, and indicate the lateral gene transfer of ail virulence genes proceeded between pathogenic and nonpathogenic Y. enterocolitica. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Molecular characterization of invasive meningococcal isolates from countries in the African meningitis belt before introduction of a serogroup A conjugate vaccine.

    Directory of Open Access Journals (Sweden)

    Dominique A Caugant

    Full Text Available BACKGROUND: The serogroup A conjugate meningococcal vaccine, MenAfriVac, was introduced in mass vaccination campaigns in December 2010 in Burkina Faso, Mali and Niger. In the coming years, vaccination will be extended to other African countries at risk of epidemics. To document the molecular characteristics of disease-causing meningococcal strains circulating in the meningitis belt of Africa before vaccine introduction, the World Health Organization Collaborating Centers on Meningococci in Europe and United States established a common strain collection of 773 isolates from cases of invasive meningococcal disease collected between 2004 and 2010 from 13 sub-Saharan countries. METHODOLOGY: All isolates were characterized by multilocus sequence typing, and 487 (62% were also analyzed for genetic variation in the surface antigens PorA and FetA. Antibiotic susceptibility was tested for part of the collection. PRINCIPAL FINDINGS: Only 19 sequence types (STs belonging to 6 clonal complexes were revealed. ST-5 clonal complex dominated with 578 (74.8% isolates. All ST-5 complex isolates were remarkably homogeneous in their PorA (P1.20,9 and FetA (F3-1 and characterized the serogroup A strains which have been responsible for most epidemics during this time period. Sixty-eight (8.8% of the 773 isolates belonged to the ST-11 clonal complex which was mainly represented by serogroup W135, while an additional 38 (4.9% W135 isolates belonged to the ST-175 complex. Forty-eight (6.2% serogroup X isolates from West Africa belonged to the ST-181 complex, while serogroup X cases in Kenya and Uganda were caused by an unrelated clone, ST-5403. Serogroup X, ST-181, emerged in Burkina Faso before vaccine introduction. CONCLUSIONS: In the seven years preceding introduction of a new serogroup A conjugate vaccine, serogroup A of the ST-5 clonal complex was identified as the predominant disease-causing strain.

  7. Whole-genome analysis of human papillomavirus genotypes 52 and 58 isolated from Japanese women with cervical intraepithelial neoplasia and invasive cervical cancer.

    Science.gov (United States)

    Tenjimbayashi, Yuri; Onuki, Mamiko; Hirose, Yusuke; Mori, Seiichiro; Ishii, Yoshiyuki; Takeuchi, Takamasa; Tasaka, Nobutaka; Satoh, Toyomi; Morisada, Tohru; Iwata, Takashi; Miyamoto, Shingo; Matsumoto, Koji; Sekizawa, Akihiko; Kukimoto, Iwao

    2017-01-01

    Human papillomavirus genotypes 52 and 58 (HPV52/58) are frequently detected in patients with cervical intraepithelial neoplasia (CIN) and invasive cervical cancer (ICC) in East Asian countries including Japan. As with other HPV genotypes, HPV52/58 consist of multiple lineages of genetic variants harboring less than 10% differences between complete genome sequences of the same HPV genotype. However, site variations of nucleotide and amino acid sequences across the viral whole-genome have not been fully examined for HPV52/58. The aim of this study was to investigate genetic variations of HPV52/58 prevalent among Japanese women by analyzing the viral whole-genome sequences. The entire genomic region of HPV52/58 was amplified by long-range PCR with total cellular DNA extracted from cervical exfoliated cells isolated from Japanese patients with CIN or ICC. The amplified DNA was subjected to next generation sequencing to determine the complete viral genome sequences. Phylogenetic analyses were performed with the whole-genome sequences to assign variant lineages/sublineages to the HPV52/58 isolates. The variability in amino acid sequences of viral proteins was assessed by calculating the Shannon entropy scores at individual amino acid positions of HPV proteins. Among 52 isolates of HPV52 (CIN1, n  = 20; CIN2/3, n  = 21; ICC, n  = 11), 50 isolates belonged to lineage B (sublineage B2) and two isolates belonged to lineage A (sublineage A1). Among 48 isolates of HPV58 (CIN1, n  = 21; CIN2/3, n  = 19; ICC, n  = 8), 47 isolates belonged to lineage A (sublineages A1/A2/A3) and one isolate belonged to lineage C. Single nucleotide polymorphisms specific for individual variant lineages were determined throughout the viral genome based on multiple sequence alignments of the Japanese HPV52/58 isolates and reference HPV52/58 genomes. Entropy analyses revealed that the E1 protein was relatively variable among the HPV52 isolates, whereas the E7, E4, and L2 proteins showed

  8. Iatrogenic aspergillus infection of the central nervous system in a pregnant woman

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    Lokuhetty Menaka

    2009-07-01

    Full Text Available A healthy postnatal woman succumbed to fulminant iatrogenic Aspergillus infection of the central nervous system, following accidental inoculation into the subarachnoid space at spinal anesthesia, during an outbreak of Aspergillus meningitis in Sri Lanka. Autopsy revealed extensive Aspergillus meningitis and culture confirmed Aspergillus fumigatus. The thalamic parenchyma in the brain was invaded by fungal hyphae producing necrotizing angitis with thrombosis, thalamic infarcts and fungal abscesses. The directional growth of fungal hyphae from the extra-luminal side of blood vessels towards the lumen favored extension from the brain parenchyma over hematogenous spread. The spinal parenchyma was resistant to fungal invasion in spite of the heavy growth within the spinal meninges and initial inoculation at spinal level. Modulation of the immune response in pregnancy with depression of selective aspects of cell-mediated immunity probably contributed to rapid spread within the subarachnoid space, to involve the brain parenchyma leading to clinical deterioration and death.

  9. New ochratoxin A or sclerotium producing species in Aspergillus section Nigri

    DEFF Research Database (Denmark)

    Samson, R.A.; Frisvad, Jens Christian

    2004-01-01

    . Aspergillus sclerotioniger was isolated from a green coffee bean and produces large yellow to red brown sclerotia and abundant ochratoxin A. The species A. homomorphus is validated. The ochratoxin A producing black aspergilli are revised. Fifteen species are provisionally accepted in Aspergillus section Nigri...... in Costa Rica and produces large pink to greyish brown sclerotia. Aspergillus lacticoffeatus was found on coffee beans in Venezuela and Indonesia, and is an effective producer of ochratoxin A. Aspergillus piperis was isolated from black ground pepper and produces large yellow to pink brown sclerotia......, four of these produce ochratoxin A. Ochratoxin A producing species of section Nigri occurring on grapes, raisins and in wine include A. carbonarius and to a lesser extent A. niger. Four species recovered from coffee, viz. A. carbonarius, A. niger, A. lacticoffeatus and A. sclerotioniger, all produce...

  10. A challenging case of invasive pulmonary aspergillosis after near-drowning: a case report and literature review.

    Science.gov (United States)

    Jenks, Jeffrey D; Preziosi, Michael

    2015-09-01

    Near-drowning, a relatively common event, is often complicated by subsequent pneumonia. While endogenous and exogenous bacteria are typical pathogens, rarely fungi are as well. We report a complicated case of invasive pulmonary aspergillosis in a 30-year-old man after a near-drowning event. We also review the medical literature for similar cases. All cases of invasive pulmonary aspergillosis after near-drowning reported in the literature involve Aspergillus fumigatus . The majority of cases involved submersion in stagnant water after a motor vehicle accident (MVA). Treatment varied considerably, with amphotericin B used in the majority of cases. Morbidity was considerable with prolonged hospitalization occurring in every case, and mortality occurring in fifty percent of the reported cases. Although a rare complication of near-drowning, invasive pulmonary aspergillosis can occur and lead to significant morbidity and mortality. After near-drowning A. fumigatus isolated from the respiratory tract should be assumed to be a true pathogen and treated accordingly.

  11. Monochrome LightCycler PCR assay for detection and quantification of five common species of Candida and Aspergillus.

    Science.gov (United States)

    Bu, Rong; Sathiapalan, Rajeev K; Ibrahim, Muna M; Al-Mohsen, Ibrahim; Almodavar, Edna; Gutierrez, Marina I; Bhatia, Kishor

    2005-03-01

    Invasive fungal pathogens, especially in immunocompromised hosts, can result in life-threatening infections. Current laboratory/radiological methods for fungal identification are time-consuming and lack sensitivity and specificity. A monochrome, multiplex, real-time PCR assay for the identification and quantification of Candida albicans, Candida krusei, Candida tropicalis, Aspergillus flavus and Aspergillus fumigatus is described here. Detection of each of these fungi was specific and demonstrated 100 % concordance with biochemical/culture identification in all 60 isolates tested. Samples from 16 febrile neutropenic patients with haematological malignancies were also analysed and the utility of the assay in clinical samples was reconfirmed without false-negative results. The sensitivity of this assay was 0.1 pg fungal genomic DNA, corresponding to three cells, for C. albicans, C. krusei, C. tropicalis and A. flavus, and 0.01 pg fungal genomic DNA, i.e. less than one cell, for A. fumigatus. The analysis allows a low-cost, simple, rapid and sensitive alternative for clinical identification and quantification of these five common fungal species.

  12. High prevalence of ceftriaxone resistance among invasive Salmonella enterica serotype Choleraesuis isolates in Thailand: The emergence and increase of CTX-M-55 in ciprofloxacin-resistant S. Choleraesuis isolates.

    Science.gov (United States)

    Luk-In, Sirirat; Chatsuwan, Tanittha; Pulsrikarn, Chaiwat; Bangtrakulnonth, Aroon; Rirerm, Ubolrat; Kulwichit, Wanla

    2018-03-23

    S. Choleraesuis is a highly invasive zoonotic pathogen that causes a serious systemic infection in humans. The emergence and increase of resistance to ceftriaxone and ciprofloxacin among S. Choleraesuis has become a serious therapeutic problem. The present study demonstrated high frequency of antimicrobial resistance in Salmonella Choleraesuis among 414 nontyphoidal Salmonella isolates from bacteremic patients in Thailand. High rates of ceftriaxone (58.3%) and ciprofloxacin (19.6%) resistances were observed in S. Choleraesuis isolates. The dissemination of the self-transferable bla CTX-M-14 -carrying IncFII s , IncFII, and IncI1 plasmids and bla CMY-2 -carrying IncA/C plasmid along with the clonal spread of bla CMY-2 -harbouring S. Choleraesuis isolates contributed to the high frequency of resistance to extended-spectrum cephalosporins (ESCs; third- and fourth-generation cephalosporins) during 2005-2007. We reported the first occurrence of ceftazidime-hydrolysing CTX-M-55 in S. Choleraesuis isolates which dramatically increased and became the most abundant CTX-M variant among ESC-resistant S. Choleraesuis isolates during 2012-2016. The spread of clone pulsotype B3 was due to the dissemination of IncA/C plasmids carrying both bla CTX-M-55 and qnrS1 among ciprofloxacin-resistant S. Choleraesuis isolates harbouring D87G in GyrA. These isolates were apparently responsible for the high rates of co-resistance to ESCs and ciprofloxacin (51.3%) during 2012-2016. This study emphasizes the importance to have an action plan to control the dissemination of antimicrobial resistance in S. Choleraesuis since this poses a threat to global health due to travel and trade in animal food products. Copyright © 2018 Elsevier GmbH. All rights reserved.

  13. A Case Report on Aspergillus lentulus Pneumonia

    Directory of Open Access Journals (Sweden)

    Muharrem Cidem

    2013-12-01

    Full Text Available Background: Aspergillus lentulus was described as a new species in 2005 but it was isolated from Turkey for the first time. Case report: A. lentulus was isolated as the cause of pneumonia from a patient who had renal transplantation 4 months ago. The patient received immunosuppressive treatment after transplantation. A. lentulus was isolated from his sputum as an agent in pneumonia developed 4 months after the transplantation. Leukocytes, blastospores, and hyphae were seen in both Gram- and Giemsa-stained smears of the sputum. The isolate was identified by using the Maren A. Klich algorithm and molecular methods and confirmed by the reference laboratory of the CBS Fungal Biodiversity Centre (The Netherlands. In the susceptibility tests of the isolate, minimal inhibitory concentrations for amphotericin B, voriconazole, posaconazole, and caspofungin were found to be 0.5 µg/mL, 0.25 µg/mL, 0.125 µg/mL, and 0.25 µg/mL, respectively. The patient recovered with voriconazole treatment (2x200 mg/day. Conclusion: The use of the molecular tests is important for identification of A. lentulus strains because they are very easily confused with A.fumigatus strains according to phenotypic characteristics.

  14. Aspergillus Polymerase Chain Reaction: Systematic Review of Evidence for Clinical Use in Comparison With Antigen Testing

    NARCIS (Netherlands)

    White, P.L.; Wingard, J.R.; Bretagne, S.; Loffler, J.; Patterson, T.F.; Slavin, M.A.; Barnes, R.A.; Pappas, P.G.; Donnelly, J.P.

    2015-01-01

    BACKGROUND: Aspergillus polymerase chain reaction (PCR) was excluded from the European Organisation for the Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) definitions of invasive fungal disease because of limited standardization and validation. The definitions are being revised.

  15. Disseminated Aspergillosis due to Aspergillus niger in Immunocompetent Patient: A Case Report

    Directory of Open Access Journals (Sweden)

    Ulku Ergene

    2013-01-01

    Full Text Available Invasive aspergillosis is a major cause of morbidity and mortality in immunocompromised patients. Many cases of pulmonary, cutaneous, cerebral, and paranasal sinus aspergillosis in immunocompetent patient were defined in literature but disseminated aspergillosis is very rare. Here we present an immunocompetent case with extrapulmonary disseminated aspergillosis due to Aspergillus niger, totally recovered after effective antifungal treatment with voriconazole.

  16. Diketopiperazines produced by an Aspergillus fumigatus Brazilian strain

    Energy Technology Data Exchange (ETDEWEB)

    Furtado, Niege A.J.C; Pupo, Monica T.; Carvalho, Ivone; Campo, Vanessa L. Campo; Bastos, Jairo K. [Sao Paulo Univ., Ribeirao Preto, SP (Brazil). Faculdade de Ciencias Farmaceuticas]. E-mail: jkbastos@fcfrp.usp.br; Duarte, Marta Cristina T. [Universidade Estadual de Campinas, SP (Brazil). Centro Pluridisciplinar de Pesquisas Quimicas, Biologicas e Agricolas

    2005-11-15

    Seven diketopiperazines, corresponding to the cycles (L)-Pro-(L)-Phe, (L)-Pro-Gly, (L)-Pro- (L)-Pro, (L)-Pro-(L)-Val, (L)-4-OH-Pro-(L)-Leu, (L)-4-OH-Pro-(L)-Phe, and (L)-Pro-(L)-Leu, were isolated from the Aspergillus fumigatus fermentation broth. The relative and absolute stereochemistries were determined on the basis of NOESY experiments and by using a modified version of Marfey's method using HPLC, respectively. (author)

  17. SPECIES OF GENUS ASPERGILLUS ON GRAPE SLOVAK ORIGIN

    OpenAIRE

    Dana Tančinová; Ľubomír Rybárik; Zuzana Mašková; Zuzana Barboráková; Soňa Felšöciová

    2014-01-01

    The aim of this study was to detect species of genus Aspergillus from wine grapes (berries, surface sterilized berries - endogenous mycobiota, from damaged berries and grape juice) of Slovak origin. We analyzed 20 samples of grapes, harvested in 2011 from various wine-growing regions. For the isolation of species we used the method of direct plating berries, surface-sterilized berries (using 0.4% freshly pre-pared chlorine), and damaged berries on DRBC (Dichloran Rose Bengal Chloramphenicol a...

  18. Workflow to study genetic biodiversity of aflatoxigenic Aspergillus spp. in Georgia, USA

    Science.gov (United States)

    Peanut seeds were sampled from the entire state of Georgia in 2014. More than 600 isolates of Aspergillus spp. were collected using modified-dichloran rose Bengal (MDRB) medium, 240 of those isolates were fingerprinted with 25 InDel markers within the aflatoxin-biosynthesis gene cluster (ABC). Clust...

  19. Polyphasic taxonomy of Aspergillus section Aspergillus (formerly Eurotium, and its occurrence in indoor environments and food

    Directory of Open Access Journals (Sweden)

    A.J. Chen

    2017-09-01

    Full Text Available Aspergillus section Aspergillus (formerly the genus Eurotium includes xerophilic species with uniseriate conidiophores, globose to subglobose vesicles, green conidia and yellow, thin walled eurotium-like ascomata with hyaline, lenticular ascospores. In the present study, a polyphasic approach using morphological characters, extrolites, physiological characters and phylogeny was applied to investigate the taxonomy of this section. Over 500 strains from various culture collections and new isolates obtained from indoor environments and a wide range of substrates all over the world were identified using calmodulin gene sequencing. Of these, 163 isolates were subjected to molecular phylogenetic analyses using sequences of ITS rDNA, partial β-tubulin (BenA, calmodulin (CaM and RNA polymerase II second largest subunit (RPB2 genes. Colony characteristics were documented on eight cultivation media, growth parameters at three incubation temperatures were recorded and micromorphology was examined using light microscopy as well as scanning electron microscopy to illustrate and characterize each species. Many specific extrolites were extracted and identified from cultures, including echinulins, epiheveadrides, auroglaucins and anthraquinone bisanthrons, and to be consistent in strains of nearly all species. Other extrolites are species-specific, and thus valuable for identification. Several extrolites show antioxidant effects, which may be nutritionally beneficial in food and beverages. Important mycotoxins in the strict sense, such as sterigmatocystin, aflatoxins, ochratoxins, citrinin were not detected despite previous reports on their production in this section. Adopting a polyphasic approach, 31 species are recognized, including nine new species. ITS is highly conserved in this section and does not distinguish species. All species can be differentiated using CaM or RPB2 sequences. For BenA, Aspergillus brunneus and A. niveoglaucus share identical

  20. Chronic invasive sinus aspergillosis in immunocompetent hosts: a geographic comparison.

    Science.gov (United States)

    Webb, Brandon J; Vikram, Holenarasipur R

    2010-12-01

    To investigate potential differences in clinical presentation, histopathology, and outcomes of chronic invasive sinus aspergillosis (CISA) based on geographic region and species of Aspergillus isolated. A retrospective analysis of published cases of CISA with a comparison of North American and worldwide cases comprised a systematic search of the English language literature. Thirty-four articles were identified detailing 15 North American and 76 global cases of CISA with cranio-cerebral extension in clinically immunocompetent patients. North American patients with CISA were older, had a more rapidly progressive course, and appeared to have higher rates of treatment failure and mortality. Anatomic distribution and presenting symptoms were similar between the two groups. North American cases were mostly due to A. fumigatus, while A. flavus was the predominant pathogen worldwide. While granulomatous inflammation was a rare observation in North American cases, it was seen in the majority of cases worldwide. CISA due to A. fumigatus was encountered in older adults, was associated with a chronic inflammatory response, an accelerated clinical course, and a trend toward treatment failure and higher mortality. Patients with A. flavus were younger, demonstrated granulomatous inflammation, and pursued an indolent, clinically responsive course. Observed differences in clinical presentation, histopathology, and outcome might involve a complex interplay between the human host, Aspergillus species, and local climatic conditions.

  1. Production of amylases by Aspergillus tamarii

    Directory of Open Access Journals (Sweden)

    Moreira Fabiana Guillen

    1999-01-01

    Full Text Available A strain of Aspergillus tamarii, a filamentous fungus isolated from soil, was able to produce both a-amylase and glucoamylase activities in mineral media supplemented with 1% (w/v starch or maltose as the carbon source. Static cultivation led to significantly higher yields than those obtained using shaking culture. The production of amylases was tolerant to a wide range of initial culture pH values (from 4 to 10 and temperature (from 25 to 42oC. Two amylases, one a-amylase and one glucoamylase, were separated by ion exchange chromatography. Both partially purified enzymes had optimal activities at pH values between 4.5 and 6.0 and were stable under acid conditions (pH 4.0-7.0. The enzymes exhibited optimal activities at temperatures between 50o and 60o C and were stable for more than ten hours at 55oC.

  2. Suppression of Aspergillus by Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Jensen, Britt Guillaume; Jelsbak, Lars; Søndergaard, Ib

    Objectives: Cystic fibrosis patients are commonly infected by Pseudomonas aeruginosa, but Aspergilli are also frequently isolated. Our aim was to examine the possible interaction between P. aeruginosa and different Aspergillus. Methods: A suspension of 106 fungal spores/ml was streaked onto WATM......, here among 2-heptyl-3-hydroxy-4-quinolone (PQS). An unidentified green pseudomonas compound was also observed. Interestingly the P. aeruginosa mutant rpoN was unable to suppress A. fumigatus, but suppressed A. flavus, A. oryzae and A. niger. However several other P. aeruginosa mutants suppressed A....... fumigatus including flif, pilA, lasR, PVDA, PQSC and rhlA mutants indicating that phenazines may be involved in the suppressed growth of A. fumigatus. All pseudomonas mutants suppressed A. oryzae, A. niger and A. flavus. Conclusions: An increase in phenazine production by P. aeruginosa may contribute...

  3. Multilocus sequence analysis of Aspergillus Sect. Nigri in dried vine fruits of worldwide origin.

    Science.gov (United States)

    Susca, Antonia; Perrone, Giancarlo; Cozzi, Giuseppe; Stea, Gaetano; Logrieco, Antonio F; Mulè, Giuseppina

    2013-07-15

    Dried vine fruits may be heavily colonized by Aspergillus species. The molecular biodiversity of an Aspergillus population (234 strains) isolated from dried vine fruit samples of worldwide origin were analyzed by investigating four housekeeping gene loci (calmodulin, β-tubulin, elongation factor 1-α, RPB2). Aspergillus Sect. Nigri was dominant and the strains were identified as A. tubingensis (138), A. awamori (38), A. carbonarius (27), A. uvarum (16) and A. niger (11). Four Aspergillus flavus strains were also identified from Chilean raisins. Two clusters closely related to the A. tubingensis species with a significant bootstrap (60% and 99%) were identified as distinct populations. Among the four loci, RPB2 showed the highest genetic variability. This is the first complete study on the worldwide distribution of black Aspergilli occurring on dried vine fruits identified by a molecular approach. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Isolation and Identification of Spoilage Fungi Associated With Rice ...

    African Journals Online (AJOL)

    The spoilage fungi isolated were Aspergillus species, Rhizopus, Penicilluim, Fusarium, Eurotium, Mucor, Geotrichum, Alternaria, Cladosporium and Actinomyces species. The predominant spoilage fungi in the grains were Aspergillus species. The populations of some spoilage fungi isolated from the grains were not high ...

  5. Isolation of novel indole diterpenes and dihydrodibenzofuran from ...

    African Journals Online (AJOL)

    Twelve secondary metabolites were earlier isolated from two marine-derived fungal strains, Aspergillus sp. AF-119 and Aspergillus sp. JQG a-6f. The objectives of this investigation were to isolate and purify natural products produced in these fungi, and identify possible drug candidates. The chemical structures of these ...

  6. Performance of Aspergillus PCR in cerebrospinal fluid for the diagnosis of cerebral aspergillosis.

    Science.gov (United States)

    Imbert, S; Brossas, J-Y; Palous, M; Joly, I; Meyer, I; Fekkar, A

    2017-11-01

    Cerebral aspergillosis is a rare but often fatal form of invasive aspergillosis that remains difficult to diagnose. The literature has shown the value of Aspergillus PCR in blood-derived samples for the diagnosis of invasive aspergillosis but provides far less information for cerebrospinal fluid (CSF) in cerebral aspergillosis. Here, we evaluated the usefulness of an Aspergillus PCR assay performed on CSF for the diagnosis of cerebral aspergillosis. This retrospective study involved 72 patients with suspected cerebral aspergillosis for a total of 88 CSF samples in whom CSF Aspergillus PCR was performed. Seventeen patients had proven/probable invasive aspergillosis according to the European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria, including 12 cases of proven/probable cerebral aspergillosis. Aspergillus PCR in CSF was positive in nine of the twelve patients with cerebral aspergillosis, i.e. 75% sensitivity. In contrast, CSF culture was positive for Aspergillus in only two patients. In the non-cerebral aspergillosis group (60 patients), PCR was positive in one patient, i.e. 98.3% specificity. In this particular population of high-risk patients with suspicion of cerebral aspergillosis, the disease incidence was 16.7%. Therefore, the positive and negative predictive values of PCR were 90% and 95.2%, respectively. The results of this study indicate that Aspergillus PCR in CSF is an interesting tool that may eliminate the need for cerebral biopsy in patients with suspected cerebral aspergillosis. Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  7. Healthy human T-Cell Responses to Aspergillus fumigatus antigens.

    Directory of Open Access Journals (Sweden)

    Neelkamal Chaudhary

    2010-02-01

    Full Text Available Aspergillus fumigatus is associated with both invasive and allergic pulmonary diseases, in different hosts. The organism is inhaled as a spore, which, if not cleared from the airway, germinates into hyphal morphotypes that are responsible for tissue invasion and resultant inflammation. Hyphae secrete multiple products that function as antigens, evoking both a protective (T(H1-T(H17 and destructive allergic (T(H2 immunity. How Aspergillus allergens (Asp f proteins participate in the development of allergic sensitization is unknown.To determine whether Asp f proteins are strictly associated with T(H2 responses, or represent soluble hyphal products recognized by healthy hosts, human T cell responses to crude and recombinant products were characterized by ELISPOT. While responses (number of spots producing IFN-gamma, IL-4 or IL-17 to crude hyphal antigen preparations were weak, responses to recombinant Asp f proteins were higher. Recombinant allergens stimulated cells to produce IFN-gamma more so than IL-4 or IL-17. Volunteers exhibited a diverse CD4+ and CD8+ T cell antigen recognition profile, with prominent CD4 T(H1-responses to Asp f3 (a putative peroxismal membrane protein, Asp f9/16 (cell wall glucanase, Asp f11 (cyclophilin type peptidyl-prolyl isomerase and Asp f22 (enolase. Strong IFN-gamma responses were reproduced in most subjects tested over 6 month intervals.Products secreted after conidial germination into hyphae are differentially recognized by protective T cells in healthy, non-atopic individuals. Defining the specificity of the human T cell repertoire, and identifying factors that govern early responses may allow for development of novel diagnostics and therapeutics for both invasive and allergic Aspergillus diseases.

  8. Healthy human T-Cell Responses to Aspergillus fumigatus antigens.

    Science.gov (United States)

    Chaudhary, Neelkamal; Staab, Janet F; Marr, Kieren A

    2010-02-17

    Aspergillus fumigatus is associated with both invasive and allergic pulmonary diseases, in different hosts. The organism is inhaled as a spore, which, if not cleared from the airway, germinates into hyphal morphotypes that are responsible for tissue invasion and resultant inflammation. Hyphae secrete multiple products that function as antigens, evoking both a protective (T(H)1-T(H)17) and destructive allergic (T(H)2) immunity. How Aspergillus allergens (Asp f proteins) participate in the development of allergic sensitization is unknown. To determine whether Asp f proteins are strictly associated with T(H)2 responses, or represent soluble hyphal products recognized by healthy hosts, human T cell responses to crude and recombinant products were characterized by ELISPOT. While responses (number of spots producing IFN-gamma, IL-4 or IL-17) to crude hyphal antigen preparations were weak, responses to recombinant Asp f proteins were higher. Recombinant allergens stimulated cells to produce IFN-gamma more so than IL-4 or IL-17. Volunteers exhibited a diverse CD4+ and CD8+ T cell antigen recognition profile, with prominent CD4 T(H)1-responses to Asp f3 (a putative peroxismal membrane protein), Asp f9/16 (cell wall glucanase), Asp f11 (cyclophilin type peptidyl-prolyl isomerase) and Asp f22 (enolase). Strong IFN-gamma responses were reproduced in most subjects tested over 6 month intervals. Products secreted after conidial germination into hyphae are differentially recognized by protective T cells in healthy, non-atopic individuals. Defining the specificity of the human T cell repertoire, and identifying factors that govern early responses may allow for development of novel diagnostics and therapeutics for both invasive and allergic Aspergillus diseases.

  9. The Pan-AC assay: a single-reaction real-time PCR test for quantitative detection of a broad range of Aspergillus and Candida species.

    Science.gov (United States)

    Basková, Lenka; Landlinger, Christine; Preuner, Sandra; Lion, Thomas

    2007-09-01

    In view of the growing incidence and the high mortality of invasive aspergillosis and candidiasis, adequate diagnostic techniques permitting timely onset of treatment are of paramount importance. More than 90 % of all invasive fungal infections in immunocompromised individuals can be attributed to Candida and Aspergillus species. To date, standardized techniques permitting rapid, sensitive and, no less importantly, economic screening for the clinically most relevant fungi are lacking. In the present report, a real-time quantitative PCR assay, developed for the detection of the most common pathogenic Candida and Aspergillus species, is described. The single-reaction PCR assay targets a judiciously selected region of the 28S subunit of the fungal rDNA gene. The unique design of the universal primer/probe system, including a pan-Aspergillus and pan-Candida (Pan-AC) hydrolysis probe, facilitates the detection of numerous Aspergillus species (e.g. Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus terreus, Aspergillus versicolor and Aspergillus nidulans) and Candida species (e.g. Candida albicans, Candida glabrata, Candida krusei, Candida tropicalis, Candida parapsilosis, Candida kefyr, Candida guilliermondii, Candida lusitaniae and Candida dubliniensis). The assay permits highly reproducible detection of 10 fg fungal DNA, which corresponds to a fraction of a fungal genome, and facilitates accurate quantification of fungal load across a range of at least five logs. Upon standardization of the technique using cultured fungal strains, the applicability in the clinical setting was assessed by investigating a series of clinical specimens from patients with documented fungal infections (n=17). The Pan-AC assay provides an attractive and economic approach to the screening and monitoring of invasive aspergillosis and candidiasis, which is readily applicable to routine clinical diagnosis.

  10. Characterization of Aspergillus section Nigri species populations in vineyard soil using droplet digital PCR.

    Science.gov (United States)

    Palumbo, J D; O'Keeffe, T L; Fidelibus, M W

    2016-12-01

    Identification of populations of Aspergillus section Nigri species in environmental samples using traditional methods is laborious and impractical for large numbers of samples. We developed species-specific primers and probes for quantitative droplet digital PCR (ddPCR) to improve sample throughput and simultaneously detect multiple species in each sample. The ddPCR method was used to distinguish Aspergillus niger, Aspergillus welwitschiae, Aspergillus tubingensis and Aspergillus carbonarius in mixed samples of total DNA. Relative abundance of each species measured by ddPCR agreed with input ratios of template DNAs. Soil samples were collected at six time points over two growing seasons from two raisin vineyards in Fresno County, California. Aspergillus section Nigri strains were detected in these soils in the range of 10 2 -10 5  CFU g -1 . Relative abundance of each species varied widely among samples, but in 52 of 60 samples, A. niger was the most abundant species, ranging from 38 to 88% of the total population. In combination with total plate counts, this ddPCR method provides a high-throughput method for describing population dynamics of important potential mycotoxin-producing species in environmental samples. This is the first study to demonstrate the utility of ddPCR as a means to quantify species of Aspergillus section Nigri in soil. This method eliminates the need for isolation and sequence identification of individual fungal isolates, and allows for greater throughput in measuring relative population sizes of important (i.e. mycotoxigenic) Aspergillus species within a population of morphologically indistinguishable species. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  11. In vivo emergence of Aspergillus terreus with reduced azole susceptibility and a Cyp51a M217I alteration

    DEFF Research Database (Denmark)

    Arendrup, Maiken C; Jensen, Rasmus; Grif, Katharina

    2012-01-01

    Azole resistance in Aspergillus terreus isolates was explored. Twenty related (MB) and 6 unrelated A. terreus isolates were included. CYP51A sequencing and RAPD genotyping was performed. Five MB isolates were itraconazole susceptible, whereas the minimum inhibitory concentrations (MICs) for 15 MB...

  12. Rapid induction of multiple resistance mechanisms in Aspergillus fumigatus during azole therapy: a case study and review of the literature.

    NARCIS (Netherlands)

    Camps, S.M.T.; Linden, J.W.M. van der; Li, Y.; Kuijper, E.J.; Dissel, J.T. van; Verweij, P.E.; Melchers, W.J.G.

    2012-01-01

    Nine consecutive isogenic Aspergillus fumigatus isolates cultured from a patient with aspergilloma were investigated for azole resistance. The first cultured isolate showed a wild-type phenotype, but four azole-resistant phenotypes were observed in the subsequent eight isolates. Four mutations were

  13. Prevalence of penicillin and erythromycin resistance among invasive Streptococcus pneumoniae isolates reported by laboratories in the southern and eastern Mediterranean region.

    Science.gov (United States)

    Borg, M A; Tiemersma, E; Scicluna, E; van de Sande-Bruinsma, N; de Kraker, M; Monen, J; Grundmann, H

    2009-03-01

    Information about the epidemiology of resistance in Streptococcus pneumoniae within southern and eastern countries of the Mediterranean region is incomplete, as reports have been sporadic and difficult to compare. Over a 36-month period, from 2003 to 2005, the ARMed project collected 1298 susceptibility test results of invasive isolates of S. pneumoniae from blood and spinal fluid cultures routinely processed within 59 participating laboratories situated in Algeria, Cyprus, Egypt, Jordan, Lebanon, Malta, Morocco, Tunisia and Turkey. Overall, 26% (335) of isolates were reported as non-susceptible to penicillin, with the highest proportions being reported from Algeria (44%) and Lebanon (40%). During the same time period, the highest proportions of pneumococci that were not susceptible to erythromycin were reported from Malta (46%) and Tunisia (39%). Proportions of dual non-susceptibility in excess of 5% were found in laboratories in Algeria, Tunisia, Lebanon, Jordan and Turkey. ARMed data on the antimicrobial resistance epidemiology of S. pneumoniae in the southern and eastern Mediterranean region provided evidence of high rates of resistance, especially to penicillin. This evidence calls for a greater focus on the identification of relevant drivers of resistance and on the implemention of effective practices in order to address the problem of resistence.

  14. Emergence of the Molds Other than Aspergillus in Immunocompromised Patients.

    Science.gov (United States)

    Arif, Sana; Perfect, John R

    2017-09-01

    Immunocompromised patients are at high risk for invasive fungal infections (IFIs); although Aspergillus remains the most common IFI caused by molds, other fungi, such as Mucorales, dematiaceous molds, and Fusarium spp, are being seen with increasing frequency. Presentations can vary, but sinopulmonary and disseminated infections are common. Our understanding of the pathogenesis of these infections is rudimentary. Fungal cultures and histopathology remain the backbone of diagnostics, as no good serologic markers are available. Polymerase chain reaction tests are being developed but currently remain investigational. Management of these infections is usually multidisciplinary, requiring surgical debridement along with antifungal therapy. Copyright © 2017 Elsevier Inc. All rights reserved.

  15. Selection arena in Aspergillus nidulans

    NARCIS (Netherlands)

    Bruggeman, J.; Debets, A.J.M.; Hoekstra, R.F.

    2004-01-01

    The selection arena hypothesis states that overproduction of zygotes-a widespread phenomenon in animals and plants-can be explained as a mechanism of progeny choice. As a similar mechanism, the ascomycetous fungus Aspergillus nidulans may overproduce dikaryotic fruit initials, hereafter called

  16. The Aspergillus Mine - publishing bioinformatics

    DEFF Research Database (Denmark)

    Vesth, Tammi Camilla; Rasmussen, Jane Lind Nybo; Theobald, Sebastian

    with the Joint Genome Institute. The Aspergillus Mine is not intended as a genomic data sharing service but instead focuses on creating an environment where the results of bioinformatic analysis is made available for inspection. The data and code is public upon request and figures can be obtained directly from...

  17. Nitrile biotransformation by Aspergillus niger

    Czech Academy of Sciences Publication Activity Database

    Šnajdrová, Radka; Kristová, Veronika; Crestia, D.; Nikolaou, K.; Kuzma, Marek; Lemaire, M.; Gallienne, E.; Bolte, J.; Bezouška, K.; Křen, Vladimír; Martínková, Ludmila

    2004-01-01

    Roč. 29, - (2004), s. 227-232 ISSN 1381-1177 R&D Projects: GA MŠk OC D25.002; GA AV ČR IAA4020213 Institutional research plan: CEZ:AV0Z5020903 Keywords : aspergillus niger * nitrile-converting enzymes * nitrile hydratase Subject RIV: EE - Microbiology, Virology Impact factor: 1.547, year: 2004

  18. Genetic relationships among strains of the Aspergillus niger aggregate

    DEFF Research Database (Denmark)

    Ferracin, L.M.; Frisvad, Jens Christian; Taniwaki, M.H.

    2009-01-01

    We analyzed the genetic relationships between 51 fungal isolates previously identified as A. niger aggregate, obtained from dried fruit samples from worldwide origin and 7 A. tubingensis obtained from Brazilian coffee beans samples. Greater fungal diversity was found in black sultanas. Aspergillus...... niger sensu stricto was the most prevalent species. It was found in all fruit substrates of all geographical origins. Based on Random Amplification of Polymorphic DNA (RAPD) and beta-tubulin sequences data two groups of A. niger were found. In spite of the small number of isolates from Group IV...

  19. Allergic Aspergillus Rhinosinusitis

    Science.gov (United States)

    Chakrabarti, Arunaloke; Kaur, Harsimran

    2016-01-01

    Allergic fungal rhinosinusitis (AFRS) is a unique variety of chronic polypoid rhinosinusitis usually in atopic individuals, characterized by presence of eosinophilic mucin and fungal hyphae in paranasal sinuses without invasion into surrounding mucosa. It has emerged as an important disease involving a large population across the world with geographic variation in incidence and epidemiology. The disease is surrounded by controversies regarding its definition and etiopathogenesis. A working group on “Fungal Sinusitis” under the International Society for Human and Animal Mycology (ISHAM) addressed some of those issues, but many questions remain unanswered. The descriptions of “eosinophilic fungal rhinosinusitis” (EFRS), “eosinophilic mucin rhinosinusitis” (EMRS) and mucosal invasion by hyphae in few patients have increased the problem to delineate the disease. Various hypotheses exist for etiopathogenesis of AFRS with considerable overlap, though recent extensive studies have made certain in depth understanding. The diagnosis of AFRS is a multi-disciplinary approach including the imaging, histopathology, mycology and immunological investigations. Though there is no uniform management protocol for AFRS, surgical clearing of the sinuses with steroid therapy are commonly practiced. The role of antifungal agents, leukotriene antagonists and immunomodulators is still questionable. The present review covers the controversies, recent advances in pathogenesis, diagnosis, and management of AFRS. PMID:29376948

  20. Allergic Aspergillus Rhinosinusitis

    Directory of Open Access Journals (Sweden)

    Arunaloke Chakrabarti

    2016-12-01

    Full Text Available Allergic fungal rhinosinusitis (AFRS is a unique variety of chronic polypoid rhinosinusitis usually in atopic individuals, characterized by presence of eosinophilic mucin and fungal hyphae in paranasal sinuses without invasion into surrounding mucosa. It has emerged as an important disease involving a large population across the world with geographic variation in incidence and epidemiology. The disease is surrounded by controversies regarding its definition and etiopathogenesis. A working group on “Fungal Sinusitis” under the International Society for Human and Animal Mycology (ISHAM addressed some of those issues, but many questions remain unanswered. The descriptions of “eosinophilic fungal rhinosinusitis” (EFRS, “eosinophilic mucin rhinosinusitis” (EMRS and mucosal invasion by hyphae in few patients have increased the problem to delineate the disease. Various hypotheses exist for etiopathogenesis of AFRS with considerable overlap, though recent extensive studies have made certain in depth understanding. The diagnosis of AFRS is a multi-disciplinary approach including the imaging, histopathology, mycology and immunological investigations. Though there is no uniform management protocol for AFRS, surgical clearing of the sinuses with steroid therapy are commonly practiced. The role of antifungal agents, leukotriene antagonists and immunomodulators is still questionable. The present review covers the controversies, recent advances in pathogenesis, diagnosis, and management of AFRS.

  1. Production and Purification of Peroxidase from Aspergillus niger.

    Directory of Open Access Journals (Sweden)

    Mohammed A. Jebor

    2017-02-01

    Full Text Available This study was conducted in the laboratories of Biology Department, College of Science, which deals with isolation and purification of peroxidase and optimization of process parameters to achieve maximum yield of peroxidase by Aspergillus niger. Solid-state fermentation of Aspergillus niger was carried out for enhanced production of peroxidase using hydrogen peroxide as the substrate of enzyme maximum activity of the enzyme was achieved under optimum growth conditions. The optimum conditions were the isolated of Aspergillus niger from soil and growth in synthetic medium, it gave high titer of peroxidase activity, the fructose as carbon source, peptone as nitrogen source, after 12 days of incubation, incubation temperature 25 °C and pH = 6.5. Peroxidase purified in four purification steps; precipitation with 70% saturation of ammonium sulfate, step of dialysis, the third by ion exchange chromatography using DEAE-Cellulose and fourth by gel filtration throughout Sephadex G-100. The specific activity of the purified enzyme was 150U/mg with 7.75 folds. The peroxidase was shown to have molecular weight of 40kDa in SDS-PAGA and about 40kDa in gel filtration.The optimum pH and temperature for peroxidase activity 7 and 35 C0 respectively.

  2. Invasive exotic plants suffer less herbivory than non-invasive exotic plants

    OpenAIRE

    Cappuccino, Naomi; Carpenter, David

    2005-01-01

    We surveyed naturally occurring leaf herbivory in nine invasive and nine non-invasive exotic plant species sampled in natural areas in Ontario, New York and Massachusetts, and found that invasive plants experienced, on average, 96% less leaf damage than non-invasive species. Invasive plants were also more taxonomically isolated than non-invasive plants, belonging to families with 75% fewer native North American genera. However, the relationship between taxonomic isolation at the family level ...

  3. Pterocarpenes elicited by Aspergillus caelatus in peanut (Arachis hypogaea) seeds.

    Science.gov (United States)

    Sobolev, Victor S; Neff, Scott A; Gloer, James B; Khan, Shabana I; Tabanca, Nurhayat; De Lucca, Anthony J; Wedge, David E

    2010-12-01

    The substituted pterocarpenes named aracarpene-1 (1) and aracarpene-2 (2) were isolated from wounded peanut seeds challenged by a strain of Aspergillus caelatus. The structures of these putative phytoalexins were determined by interpretation of NMR and MS data. The aracarpenes were investigated for their antifungal and antibacterial activities as well as antioxidant, anti-inflammatory, and cytotoxic activities in mammalian cells. Aracarpene-2 demonstrated high antibacterial properties against tested gram-positive and gram-negative bacteria, whereas aracarpene-1 displayed low antibacterial properties against the same bacteria. Both compounds had no antifungal activity against Aspergillus flavus. Together with peanut stilbenoids that are also produced in the challenged seeds, these compounds may represent a class of low-molecular weight peanut metabolites with a defensive role(s) against pathogenic microorganisms. Published by Elsevier Ltd.

  4. Antibiotic Sensitivity Pattern of Microorganisms Isolated from ...

    African Journals Online (AJOL)

    ., Pseudomonas sp., Corynebacterium sp., Micrococcus sp. and four fungal species namely; Aspergillus spp., Fusarium sp., Mucor sp. and Penicillium spp. were identified. Majority of the bacteria were isolated from fish samples from Benin.

  5. The role of interleukin-1 family members in the host defence against Aspergillus fumigatus.

    Science.gov (United States)

    Gresnigt, Mark S; van de Veerdonk, Frank L

    2014-12-01

    The interleukin (IL)-1 family consists of 11 members, which all play significant roles in regulating inflammatory responses in the host. IL-1α and IL-1β exert potent pro-inflammatory effects and are key players in the recruitment of neutrophils to the site of inflammation. Protective anti-Aspergillus host responses during the early stages of invasive aspergillosis are critically dependent on neutrophil recruitment, and several lines of evidence support that there is an important role for IL-1 in this process. However, IL-1-mediated inflammation needs to be tightly regulated, since uncontrolled inflammation can result in inflammatory pathology and thereby be detrimental for the host. Aspergillus-induced IL-1-mediated inflammation could therefore be amendable for IL-1 blockade under specific circumstances. This review describes the current understanding of the role of IL-1 family members in the host response against Aspergillus fumigatus and highlights the importance of balanced IL-1 responses in aspergillosis.

  6. Biodiversity of Aspergillus section Nigri populations in Argentinian vineyards and ochratoxin A contamination.

    Science.gov (United States)

    Chiotta, María L; Ponsone, María L; Sosa, Débora M; Combina, Mariana; Chulze, Sofía N

    2013-12-01

    Aspergillus section Nigri are described as the main source of ochratoxin A (OTA) contamination in grapes and wine worldwide. The knowledge of the factors affecting grape contamination by species included in this section and OTA production is essential to be able to reduce their presence, not only to improve wine quality, but also to maintain their safety. Therefore, the aims of this study were to determine the incidence of Aspergillus section Nigri species harvested in different grape-growing regions from Argentina, their ability to produce OTA, to correlate with meteorological conditions and geographical coordinates with their prevalence and to evaluate the OTA natural occurrence in grapes and wines. The morphological identification showed that Aspergillus niger aggregate species were the most prevalent ones, followed by Aspergillus carbonarius and Aspergillus uniseriate. These populations were confirmed through using AFLP markers and sequencing and, Aspergillus tubingensis was separated from A. niger aggregate. Climatic factors, altitude, longitude and latitude have influenced on the distribution of species included in the section. A. carbonarius and A. niger were OTA producers but differed in their OTA producing ability. Temperature was the factor which influenced the most over the highest incidence of A. carbonarius in La Rioja and San Juan regions. The trellis system in vineyards and drip irrigation also influenced the species isolation. The OTA levels detected in grapes and wines were low, but grape variety was more important in susceptibility to fungal infection and OTA levels. Copyright © 2013 Elsevier Ltd. All rights reserved.

  7. Effect of Aspergillus versicolor strain JASS1 on low density polyethylene degradation

    Science.gov (United States)

    Gajendiran, A.; Subramani, S.; Abraham, J.

    2017-11-01

    Low density polyethylene (LDPE) waste disposal remains one of the major environmental concerns faced by the world today. In past decades, major focus has been given to enhance the biodegradation of LDPE by microbial species. In this present study, Aspergillus versicolor with the ability to degrade LDPE was isolated from municipal landfill area using enrichment technique. Based on 18S rRNA gene sequencing confirmed its identity as Aspergillus versicolor. The biodegradation study was carried out for 90 d in M1 medium. The degradation behaviour of LDPE films by Aspergillus versicolor strain JASS1 were confirmed by weight loss, CO2 evolution, Scanning electron microscopy (SEM) analysis, Atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FTIR) technique. From current investigation, it can be concluded that our isolated strain JASS1 had the potential to degrade LDPE films and it can be useful in solving the problem caused by polyethylene in the environment.

  8. [Invasive aspergillosis after near-drowning: case reports and review of the literature].

    Science.gov (United States)

    Li, Pei; Cao, E-hong; Zhao, Bei-lei; Sun, Hui-ming; Li, Miao-miao; Xu, Jin; Song, Yong; Shi, Yi

    2011-09-01

    The purpose of this study was to analyze the clinical presentations and disease courses of invasive aspergillosis (IA) in patients after near-drowning. The clinical data of 3 cases of invasive aspergillosis after near-drowning from Oct. 2005 to Aug. 2010 in this hospital were retrospectively analyzed, and the related literature was reviewed. There were 1 male and 2 female patients, aged from 18 to 72 years. All of them had been immunocompetent before drowning. Two patients drowned because of traffic accident, and 1 fell in sewage by accident. All of the 3 patients were intubated because of acute respiratory failure, and received broad-spectrum antibiotic therapy. One had transient leucopenia, and 2 patients received glucocorticoid therapy. The condition of the 3 cases deteriorated 9 to 11 days after near-drowning. Aspergillus was isolated from sputum samples of 2 patients at the same time. Thoracic CT findings included multiple nodules, consolidation and cavity formation. Multiple abscesses in cerebral parenchyma were found in 1 patient with invasive cerebral aspergillosis. One patient died, whose lungs, cerebral parenchyma, myocardium and kidney were all infected by aspergillus. The other 2 patients, whose infection limited to the lungs, had a positive prognosis. Using the terms "aspergillosis" and "near-drowning" a PUBMED search yielded 7 articles, published between 1984 and 2010. Using the terms "invasive pulmonary aspergillosis" and "near-drowning", searching Wangfang data and CHED data, encompass 1 article, published in 2009. In all of the 8 articles, there are 5 final diagnosis cases and 3 clinical diagnosis cases. IA was very rare in immunocompetent hosts but had been reported in previously healthy individuals after near-drowning. Aspergillosis might develop 1 to 2 weeks after near-drowning, and the prognosis was poor in patients with central nervous system involvement.

  9. Lipoxygenase Activity Accelerates Programmed Spore Germination in Aspergillus fumigatus

    Directory of Open Access Journals (Sweden)

    Gregory J. Fischer

    2017-05-01

    Full Text Available The opportunistic human pathogen Aspergillus fumigatus initiates invasive growth through a programmed germination process that progresses from dormant spore to swollen spore (SS to germling (GL and ultimately invasive hyphal growth. We find a lipoxygenase with considerable homology to human Alox5 and Alox15, LoxB, that impacts the transitions of programmed spore germination. Overexpression of loxB (OE::loxB increases germination with rapid advance to the GL stage. However, deletion of loxB (ΔloxB or its signal peptide only delays progression to the SS stage in the presence of arachidonic acid (AA; no delay is observed in minimal media. This delay is remediated by the addition of the oxygenated AA oxylipin 5-hydroxyeicosatetraenoic acid (5-HETE that is a product of human Alox5. We propose that A. fumigatus acquisition of LoxB (found in few fungi enhances germination rates in polyunsaturated fatty acid-rich environments.

  10. Dereplication Guided Discovery of Secondary Metabolites of Mixed Biosynthetic Origin from Aspergillus aculeatus

    DEFF Research Database (Denmark)

    Petersen, Lene Maj; Hoeck, Casper; Frisvad, Jens Christian

    2014-01-01

    Investigation of the chemical profile of the industrially important black filamentous fungus Aspergillus aculeatus by UHPLC-DAD-HRMS and subsequent dereplication has led to the discovery of several novel compounds. Isolation and extensive 1D and 2D NMR spectroscopic analyses allowed for structura...... Candida albicans, however all showed only weak or no activity. Aspergillus aculeatus IBT 21030 was additionally shown to be capable of producing sclerotia. Examination of the sclerotia revealed a highly regulated production of metabolites in these morphological structures....

  11. Culture-Based Methods and Molecular Tools for Azole-Resistant Aspergillus fumigatus Detection in a Belgian University Hospital

    OpenAIRE

    Montesinos, I.; Argudín, M. A.; Hites, M.; Ahajjam, F.; Dodémont, M.; Dagyaran, C.; Bakkali, M.; Etienne, I.; Jacobs, F.; Knoop, C.; Patteet, S.; Lagrou, K.

    2017-01-01

    Azole-resistant Aspergillus fumigatus is an increasing worldwide problem with major clinical implications. Surveillance is warranted to guide clinicians to provide optimal treatment to patients. To investigate azole resistance in clinical Aspergillus isolates in our institution, a Belgian university hospital, we conducted a laboratory-based surveillance between June 2015 and October 2016. Two different approaches were used: a prospective culture-based surveillance using VIPcheck on unselected...

  12. SNaPAfu: a novel single nucleotide polymorphism multiplex assay for aspergillus fumigatus direct detection, identification and genotyping in clinical specimens.

    Directory of Open Access Journals (Sweden)

    Rita Caramalho

    Full Text Available OBJECTIVE: Early diagnosis of invasive aspergillosis is essential for positive patient outcome. Likewise genotyping of fungal isolates is desirable for outbreak control in clinical setting. We designed a molecular assay that combines detection, identification, and genotyping of Aspergillus fumigatus in a single reaction. METHODS: To this aim we combined 20 markers in a multiplex reaction and the results were seen following mini-sequencing readings. Pure culture extracts were firstly tested. Thereafter, Aspergillus-DNA samples obtained from clinical specimens of patients with possible, probable, or proven aspergillosis according to European Organization for the Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG criteria. RESULTS: A new set of designed primers allowed multilocus sequence typing (MLST gene amplification in a single multiplex reaction. The newly proposed SNaPAfu assay had a specificity of 100%, a sensitivity of 89% and detection limit of 1 ITS copy/mL (∼0.5 fg genomic Aspergillus-DNA/mL. The marker A49_F was detected in 89% of clinical samples. The SNaPAfu assay was accurately performed on clinical specimens using only 1% of DNA extract (total volume 50 µL from 1 mL of used bronchoalveolar lavage. CONCLUSIONS: The first highly sensitive and specific, time- and cost-economic multiplex assay was implemented that allows detection, identification, and genotyping of A. fumigatus strains in a single amplification followed by mini-sequencing reaction. The new test is suitable to clinical routine and will improve patient management.

  13. Transcriptional profiling of Aspergillus niger

    OpenAIRE

    Veen, van der, D.

    2009-01-01

    The industrially important fungus Aspergillus niger feeds naturally on decomposing plant material, of which a significant proportion is lipid. Examination of the A. niger genome sequence suggested that all proteins required for metabolic conversion of lipids are present, including 63 predicted lipases. In contrast to polysaccharide-degrading enzyme networks, not much is known about the signaling and regulatory processes that control lipase expression and activity in fungi. This project was ai...

  14. Hyphal heterogeneity in Aspergillus niger

    OpenAIRE

    de Bekker, A.M.

    2011-01-01

    Mycelial fungi use hyphae to colonize substrates. These hyphae secrete enzymes that convert complex polymers into breakdown products that can be taken up to serve as nutrients. Using GFP as a reporter it has been shown that exploring hyphae of Aspergillus niger are heterogenic with respect to expression of the glucoamylase gene glaA; some hyphae strongly express the glucoamylase gene glaA, while others express it lowly. This was a surprising finding considering the fact that all hyphae were e...

  15. Germination of Aspergillus niger conidia

    OpenAIRE

    Hayer, Kimran

    2014-01-01

    Aspergillus niger is a black-spored filamentous fungus that forms asexual spores called conidospores (‘conidia’). Germination of conidia, leading to the formation of hyphae, is initiated by conidial swelling and mobilisation of endogenous carbon and energy stores, followed by polarisation and emergence of a hyphal germ tube. These morphological and biochemical changes which define the model of germination have been studied with the aim of understanding how conidia sense and utilise different...

  16. Regulatory processes in Aspergillus niger

    OpenAIRE

    Poulsen, Lars; Thykær, Jette; Eliasson Lantz, Anna

    2012-01-01

    Filamentous fungi are extensively used in the fermentation industry for synthesis of numerous products. One of the most important, is the fungus Aspergillus niger, used industrially for production of organic acids, and homologous as well as heterologous enzymes. This fungus has numerous of advantages, including tolerance for low pH, which is important for acid production. Furthermore, it has the capability of metabolizing a wide variety of carbon sources, possesses an exceptional efficient pr...

  17. New restriction fragment length polymorphism (RFLP) markers for Aspergillus fumigatus.

    Science.gov (United States)

    Semighini, C P; Delmas, G; Park, S; Amstrong, D; Perlin, D; Goldman, G H

    2001-07-01

    In this study, we isolated and tested restriction fragment length polymorphism (RFLP) markers for Aspergillus fumigatus based on PCR products amplified by the random amplified polymorphic DNA (RAPD) primer R108. Four DNA fragments, Afd, Af5, Af4, and Af4A, were amplified. Fragments Afd and Af5 were 85% and 88% identical at the DNA level to part of the Afut1 retrotransposon from A. fumigatus. Fragment Af4A is a duplication of fragment Af4 and both showed similarity at the amino acid level with endonucleases from other fungal retrotransposons. We used both RAPD with primer R108 and RFLP assays with Afut1, Afd, and Af4A, to determine the genetic relatedness of clinical isolates of A. fumigatus isolated sequentially from four patients colonized with A. fumigatus. The combination of these different methods suggested that the isolates infecting the four patients were not identical.

  18. Non-invasive Prenatal Diagnosis of Chromosomal Aneuploidies and Microdeletion Syndrome Using Fetal Nucleated Red Blood Cells Isolated by Nanostructure Microchips.

    Science.gov (United States)

    Feng, Chun; He, Zhaobo; Cai, Bo; Peng, Jianhong; Song, Jieping; Yu, Xuechen; Sun, Yue; Yuan, Jing; Zhao, Xingzhong; Zhang, Yuanzhen

    2018-01-01

    Detection of detached fetal nucleated red blood cells (fNRBCs) in the maternal peripheral blood may serve as a prospective testing method competing with the cell-free DNA, in non-invasive prenatal testing (NIPT). Methods: Herein, we introduce a facile and effective lab-on-a-chip method of fNRBCs detection using a capture-releasing material that is composed of biotin-doped polypyrrole nanoparticles. To enhance local topographic interactions between the nano-components and fNRBC, a specific antibody, CD147, coated on the nanostructured substrate led to the isolation of fNRBCs from maternal peripheral blood. Subsequently, an electrical system was employed to release the captured cells using 0.8 V for 15 s. The diagnostic application of fNRBCs for fetal chromosomal disorders (Trisomy 13/21/18/X syndrome, microdeletion syndrome) was demonstrated. Results: Cells captured by nanostructured microchips were identified as fNRBCs. Twelve cases of chromosomal aneuploidies and one case of 18q21 microdeletion syndrome were diagnosed using the fNRBCs released from the microchips. Conclusion: Our method offers effective and accurate analysis of fNRBCs for comprehensive NIPT to monitor fetal cell development.

  19. Melanin is an essential component for the integrity of the cell wall of Aspergillus fumigatus conidia

    Directory of Open Access Journals (Sweden)

    Georgeault Sonia

    2009-08-01

    Full Text Available Abstract Background Aspergillus fumigatus is the most common agent of invasive aspergillosis, a feared complication in severely immunocompromised patients. Despite the recent commercialisation of new antifungal drugs, the prognosis for this infection remains uncertain. Thus, there is a real need to discover new targets for therapy. Particular attention has been paid to the biochemical composition and organisation of the fungal cell wall, because it mediates the host-fungus interplay. Conidia, which are responsible for infections, have melanin as one of the cell wall components. Melanin has been established as an important virulence factor, protecting the fungus against the host's immune defences. We suggested that it might also have an indirect role in virulence, because it is required for correct assembly of the cell wall layers of the conidia. Results We used three A. fumigatus isolates which grew as white or brown powdery colonies, to demonstrate the role of melanin. Firstly, sequencing the genes responsible for biosynthesis of melanin (ALB1, AYG1, ARP1, ARP2, ABR1 and ABR2 showed point mutations (missense mutation, deletion or insertion in the ALB1 gene for pigmentless isolates or in ARP2 for the brownish isolate. The isolates were then shown by scanning electron microscopy to produce numerous, typical conidial heads, except that the conidia were smooth-walled, as previously observed for laboratory mutants with mutations in the PKSP/ALB1 gene. Flow cytometry showed an increase in the fibronectin binding capacity of conidia from mutant isolates, together with a marked decrease in the binding of laminin to the conidial surface. A marked decrease in the electronegative charge of the conidia and cell surface hydrophobicity was also seen by microelectrophoresis and two-phase partitioning, respectively. Ultrastructural studies of mutant isolates detected considerable changes in the organisation of the conidial wall, with the loss of the outermost

  20. Aspergillus in endodontic infection near the maxillary sinus

    Directory of Open Access Journals (Sweden)

    Cinthya Cristina Gomes

    2015-10-01

    Full Text Available ABSTRACT INTRODUCTION: Diseases of the maxillary sinus have been associated with dental roots near the maxillary sinus that have undergone endodontic treatment. OBJECTIVE: To investigate the presence of filamentous fungi in patients with dental roots near the maxillary sinus who had apical periodontitis treated endodontically, and to alert practitioners that this could be a possible avenue of contamination of the sinus in patients who develop maxillary sinus infection. METHODS: Cross-sectional study in 60 palatal roots of the first maxillary molars near the maxillary sinus, that underwent endodontic treatment for apical periodontitis. After removal of the filling material, dentin shavings were collected and placed in test tubes containing Sabouraud dextrose agar and chloramphenicol. The phenotype was determined by macroscopic and microscopic examination of the colonies. For polymerase chain reaction, the primers ITS-5 and ITS-4 were used. The sequences obtained were compared with those deposited at GenBank using the Basic Local Alignment Search Tool program. RESULTS: Filamentous fungi were isolated from 6 of 60 canals (10%:Aspergillus niger (6.7%, Aspergillus versicolor (1.6%, and Aspergillus fumigatus(1.6%. CONCLUSION: Root canals near the maxillary sinus with endodontic treatment and apical periodontitis may exhibit positive cultures for filamentous fungi. Interested professionals should be alert, because these microorganisms have pathogenic characteristics that can cause disease of odontogenic origin in the maxillary sinus.

  1. Cytokine responses and regulation of interferon-gamma release by human mononuclear cells to Aspergillus fumigatus and other filamentous fungi.

    NARCIS (Netherlands)

    Warris, A.; Netea, M.G.; Verweij, P.E.; Gaustad, P.; Kullberg, B.J.; Weemaes, C.M.R.; Abrahamsen, T.G.

    2005-01-01

    There is substantial evidence that the production of proinflammatory cytokines is important in host resistance to invasive aspergillosis. Knowledge of the host response towards other filamentous fungi is scarce, as most studies have focused on Aspergillus fumigatus. In addition, interferon-gamma

  2. Aspergillus species and other molds in respiratory samples from patients with cystic fibrosis:

    DEFF Research Database (Denmark)

    Mortensen, Klaus Leth; Jensen, Rasmus Hare; Johansen, Helle Krogh

    2011-01-01

    Respiratory tract colonization by molds in patients with cystic fibrosis (CF) were analyzed, with particular focus on the frequency, genotype, and underlying mechanism of azole resistance among Aspergillus fumigatus isolates. Clinical and demographic data were also analyzed. A total of 3,336 resp...

  3. Production of bio-ethanol from corncobs using Aspergillus niger and ...

    African Journals Online (AJOL)

    Maize is the most abundant cereal grown in Ghana and is accompanied by enormous amount of agrowastes of which corncobs form 30%. This agrowaste which is currently under utilized was used to produce bio-ethanol. Aspergillus niger isolated from soil sampled from Ejura farms was used to hydrolyze the corncobs into ...

  4. The kinetics of glucose production from rice straw by Aspergillus niger

    African Journals Online (AJOL)

    STORAGESEVER

    2008-06-03

    Jun 3, 2008 ... 3Department of Chemical Engineering, University of Maiduguri, Maiduguri, Nigeria. Accepted 2 May, 2008. In this investigation, glucose was produced from rice straw using cells of Aspergillus niger, isolated from maize grain. Glucose yield was found to increase from 43 to 87% as the rice straw particle size.

  5. Intraspecific competition during infection by Aspergillus flavus is influenced by plant host species

    Science.gov (United States)

    Communities of Aspergillus flavus are composed of diverse genotypes that collectively influence incidence and severity of crop aflatoxin contamination. Isolates vary in competitive ability on maize, but empirical data on the extent to which host-specific influences determine outcomes of competition ...

  6. Keratitis caused by the recently described new species Aspergillus brasiliensis: two case reports

    Directory of Open Access Journals (Sweden)

    Vágvölgyi Csaba

    2010-02-01

    Full Text Available Abstract Introduction Human infections caused by Aspergillus brasiliensis have not yet been reported. We describe the first two known cases of fungal keratitis caused by Aspergillus brasiliensis. Case presentations A 49-year-old Indian Tamil woman agricultural worker came with pain and defective vision in the right eye for one month. Meanwhile, a 35-year-old Indian Tamil woman presented with a history of a corneal ulcer involving the left eye for 15 days. The fungal strains isolated from these two cases were originally suspected to belong to Aspergillus section Nigri based on macro- and micromorphological characteristics. Molecular identification revealed that both isolates represent A. brasiliensis. Conclusion The two A. brasiliensis strains examined in this study were part of six keratitis isolates from Aspergillus section Nigri, suggesting that this recently described species may be responsible for a significant proportion of corneal infections caused by black Aspergilli. The presented cases also indicate that significant differences may occur between the severities of keratitis caused by individual isolates of A. brasiliensis.

  7. Regulation and control of L-arabinose catabolism in Aspergillus niger

    NARCIS (Netherlands)

    Groot, de M.J.L.

    2005-01-01

    This thesis describes studies on the biochemical properties and regulation of L-arabinose metabolism and arabinan degrading enzymes of Aspergillus niger. We focused on the investigation of the catabolic pathway, firstly by isolating pathway specific regulatory mutants using a newly developed

  8. Application of Aspergillus niger SA1 for the enhanced bioremoval of ...

    African Journals Online (AJOL)

    Biological remediation is always envisaged as cost effective and eco-friendly for the treatment of recalcitrant dyes and effluents. Aspergillus niger SA1, a brown rot fungi, isolated from storage pond of textile wastewater, showed a great mineralizing ability for azo dyes, acid red (AR) 151 and orange (Or) II. Decolorization ...

  9. Characterization of the fumonisin B2 biosynthetic gene cluster in Aspergillus niger and A. awamori.

    Science.gov (United States)

    Aspergillus niger and A. awamori strains isolated from grapes cultivated in Mediterranean basin were examined for fumonisin B2 (FB2) production and presence/absence of sequences within the fumonisin biosynthetic gene (fum) cluster. Presence of 13 regions in the fum cluster was evaluated by PCR assay...

  10. Conversion of fusaric acid to fusarinol by Aspergillus niger: A detoxification reaction

    Science.gov (United States)

    The fungus Fusarium oxysporum causes wilt diseases of plants and produces a potent phytotoxin fusaric acid (FA) which is also toxic to many microorganisms. An Aspergillus strain with high tolerance to FA was isolated from soil. HPLC analysis of culture filtrates from A. niger grown with the addition...

  11. Tools for assessing kinship, population structure, phylogeography, and interspecific hybridization in Asian carps invasive to the Mississippi River, USA: isolation and characterization of novel tetranucleotide microsatellite DNA loci in silver carp Hypophthalmichthys molitrix

    Science.gov (United States)

    King, T.L.; Eackles, M.S.; Chapman, D.C.

    2011-01-01

    We document the isolation and characterization of novel tetranucleotide microsatellite DNA markers for the invasive silver carp Hypophthalmichthys molitrix and provide the results of cross-species amplification for three additional invasive carp species: bighead (H. nobilis), grass (Ctenopharyngodon idella) and black (Mylopharyngodon piceus). In the target species these markers yielded levels of allelic diversity (average 4.4 alleles/locus) and heterozygosity (average 54.7%) sufficient to: (1) provide unique multilocus genotypes; (2) delineate kinship relationships; (3) differentiate populations/species; (4) estimate effective population sizes; and (5) provide unique demographic perspectives for control or eradication. Currently these markers are being utilized to determine the degree of introgressive hybridization between H. molitrix and H. nobilis, to quantify gene flow between different sub-basins established in the central United States, and to assess the demographic status of sub-basin groups. This information will be critically important in the management/control of these invasive species.

  12. Prevalence, persistence, and phenotypic variation of Aspergillus fumigatus in the outdoor environment in Manchester, UK, over a 2-year period.

    Science.gov (United States)

    Alshareef, Fadwa; Robson, Geoffrey D

    2014-05-01

    Aspergillus fumigatus, an opportunistic fungal pathogen that causes invasive aspergillosis in immunosuppressed patients, is considered to be the world's most dangerous mould. It is widely distributed in the environment, and airborne asexual conidia serve as the main mode of transport for pulmonary lung infection. It is important to monitor seasonal airborne conidia levels when assessing the risk of acquiring this infection. In this study, air was sampled for total viable fungal spores and viable A. fumigatus conidia monthly over a 2-year period (2009 and 2010) close to Manchester, UK, city center. Total viable airborne fungal counts varied seasonally, peaking in the summer and autumn for both years and reaching levels of approximately 1100-1400 colony-forming units (CFU)/m(3); counts were strongly positively correlated to mean temperature (R(2) = 0.697). By contrast, A. fumigatus viable airborne counts were not seasonally associated; persistent low levels were between 3 and 20 CFU/m(3) and were not correlated with mean temperature (R(2) = 0.018). A total of 220 isolates of A. fumigatus were recovered on potato dextrose agar (PDA) at 45°C, and internal transcribed spacer sequencing and restriction digestion of a partial polymerase chain reaction amplicon of the β-tubulin gene (benA) of 34 randomly selected isolates were used to confirm the isolates as A. fumigatus. When the colony radial growth rates (Kr) were determined, the highest rates were observed on PDA, followed by Vogel's medium supplemented with phosphatidylcholine and Vogel's medium alone. Clinical isolates had a significantly higher mean colony Kr on PDA compared with environmental isolates.

  13. The radiological spectrum of invasive aspergillosis in children: a 10-year review

    International Nuclear Information System (INIS)

    Thomas, Karen E.; Owens, Catherine M.; Veys, Paul A.; Novelli, Vas; Costoli, Vera

    2003-01-01

    Invasive aspergillosis is an uncommon but life-threatening event in the immunocompromised child. Attempts at fungal isolation are often unrewarding and a high index of radiological suspicion is essential in the early diagnosis of infected children. To document the radiological spectrum of disease in invasive aspergillosis in the paediatric population. A retrospective review of the imaging performed in 27 consecutive patients (age 7 months to 18 years) with documented invasive Aspergillosis encountered over a 10-year period at a single institution. Radiographic findings of pulmonary disease (20 patients) included segmental and multilobar consolidation, perihilar infiltrates, multiple small nodules, peripheral nodular masses and pleural effusions. No cavitating lesions were seen on CXR. Small cavitating nodules were present on CT in two of eight children. Chest wall disease was particularly associated with underlying chronic granulomatous disease. Disseminated disease manifested as osteomyelitis (n=5), cerebral (n=3), oesophageal (n=1), hepatic (n=2), renal (n=2) and cutaneous (n=5) involvement. Imaging findings are discussed. Twelve patients (44%) subsequently died from Aspergillus-related complications. Invasive aspergillosis presents with a wide variety of radiographic findings involving multiple organ systems. Respiratory findings are varied but often non-specific, and a high index of suspicion is necessary in immunocompromised patients. In contrast to adult disease, the incidence of cavitation of pulmonary lesions appears low. (orig.)

  14. Sporulation inhibited secretion in Aspergillus niger

    NARCIS (Netherlands)

    Krijgsheld, P.

    2013-01-01

    Aspergillus niger is abundantly found in nature. It degrades dead material of plants and animals but can also be a pathogen of these organisms. Aspergillus niger is also important for mankind because it is one of the main organisms used for the industrial production of enzymes. These enzymes are

  15. Identification of a putative Crf splice variant and generation of recombinant antibodies for the specific detection of Aspergillus fumigatus.

    Directory of Open Access Journals (Sweden)

    Mark Schütte

    Full Text Available BACKGROUND: Aspergillus fumigatus is a common airborne fungal pathogen for humans. It frequently causes an invasive aspergillosis (IA in immunocompromised patients with poor prognosis. Potent antifungal drugs are very expensive and cause serious adverse effects. Their correct application requires an early and specific diagnosis of IA, which is still not properly achievable. This work aims to a specific detection of A. fumigatus by immunofluorescence and the generation of recombinant antibodies for the detection of A. fumigatus by ELISA. RESULTS: The A. fumigatus antigen Crf2 was isolated from a human patient with proven IA. It is a novel variant of a group of surface proteins (Crf1, Asp f9, Asp f16 which belong to the glycosylhydrolase family. Single chain fragment variables (scFvs were obtained by phage display from a human naive antibody gene library and an immune antibody gene library generated from a macaque immunized with recombinant Crf2. Two different selection strategies were performed and shown to influence the selection of scFvs recognizing the Crf2 antigen in its native conformation. Using these antibodies, Crf2 was localized in growing hyphae of A. fumigatus but not in spores. In addition, the antibodies allowed differentiation between A. fumigatus and related Aspergillus species or Candida albicans by immunofluorescence microscopy. The scFv antibody clones were further characterized for their affinity, the nature of their epitope, their serum stability and their detection limit of Crf2 in human serum. CONCLUSION: Crf2 and the corresponding recombinant antibodies offer a novel approach for the early diagnostics of IA caused by A. fumigatus.

  16. Susceptibilidad a antimicrobianos en aislamientos de Streptococcus pneumoniae invasor en Colombia Susceptibility to antimicrobial agents in isolates of invasive Streptococcus pneumoniae in Colombia

    Directory of Open Access Journals (Sweden)

    Aura Lucía Leal

    1999-03-01

    pneumoniae that caused invasive disease diagnosed in children under the age of 5 in Colombia between 1994 and 1996, as well as to establish the distribution of the capsular types of the resistant isolates. The analysis was done using 324 isolates obtained during the performance of the National Serotyping Protocol for S. pneumoniae carried out in Bogotá, Medellín, and Cali, Colombia, between July 1994 and March 1996. Of the 324 isolates, 119 (36.7% showed diminished susceptibility to at least one antimicrobial agent, including 39 (12% that showed diminished susceptibility to penicillin. Of these 39 resistant to penicillin, 29 showed intermediate resistance and 10 showed high resistance. Nine isolates (2.8% showed resistance to ceftriaxone, 80 (24.7% to the combination of trimethoprim and sulfamethoxazole (TMS, 49 (15.1% to chloramphenicol, and 31 (9.6% to erythromycin. Resistance to two antimicrobial agents was observed in 31 isolates (9.6%; multiple resistance was found in 22 (6.7%. These 22 multiresistant isolates all showed resistance to TMS. The most frequent associations were penicillin, TMS, and erythromycin (5 cases; penicillin, chloramphenicol, TMS, and erythromycin (4 cases; penicillin, ceftriaxone, chloramphenicol, and TMS (3 cases; and penicillin, ceftriaxone, chloramphenicol, TMS, and erythromycin (3 cases. The most frequent serotypes in the penicillin-resistant isolates were: 23F (53.8%, 14 (25.6%, 6B (7.7%, 9V (5.1%, 19F (5.1%, and 34 (2.6%. The most frequent serotypes in the isolates resistant to antimicrobial agents other than penicillin were: 5 (37.5%, 23F (7.5%, 14 (18.8%, and 6B (13.8%. This difference in the distribution of the serotypes was statistically significant (P < 0.0001. The study results indicate the need to maintain active surveillance of antibiotic susceptibility patterns in order to avoid resistance in S. pneumoniae and to provide timely in formation to change practices regarding prescribing and consuming antimicrobial agents.

  17. Rapidly progressing dual infection with Aspergillus and Rhizopus: when soil inhabitants become deadly invaders.

    Science.gov (United States)

    Bhagat, Milind; Rapose, Alwyn

    2016-12-08

    We present a case report of a 61-year-old patient with acute pulmonary and cerebral infections with Aspergillus and Rhizopus. The only risk factor for invasive fungal disease was high-dose corticosteroids used to treat her chronic obstructive pulmonary disease exacerbation. She had rapid progression and succumbed to her infections within 2 weeks of diagnosis in spite of aggressive antifungal therapy and surgery. To the best of our knowledge, this is the first reported case of rapidly fatal dual infection with Aspergillus and Rhizopus Our case highlights the role of high-dose corticosteroids as a risk factor for invasive fungal disease in patients without traditional risk factors like haematological malignancies, solid organ transplantation or uncontrolled diabetes. 2016 BMJ Publishing Group Ltd.

  18. Detection of Aflatoxin Producing Aspergillus flavus in Post-harvest Contaminated Vigna ungulculata Seeds

    Directory of Open Access Journals (Sweden)

    Ajay Kumar Gautam

    2014-07-01

    Full Text Available The present study was carried out with a specific objective to study postharvest spoilage of Lobhiya (Vigna unguiculata seeds contaminated with Aspergillus flavus. Infected seeds were collected and cultured on potato dextrose agar (PDA media, at 25±2 °C. Aspergillus flavus isolates were primarily characterized by its morphological and microscopic characteristics. Collected fungal isolates were also screened for their afaltoxigenic nature on preliminary basis and at molecular level. For preliminary screening, 5 mm disc of fungal culture was soaked with few drops of liquid ammonia. Color change from yellow pigment to plum-red with different intensities showed the mycotoxic nature of the fungus. DNA from fungal isolates was isolated and amplified using PCR with aflatoxin specific primers, apa-2, ver-1 and omt-1. Amplicons of 1032 bp, 895 bp and 596 bp were obtained in most of the isolates regardless of primer set used which was useful to differentiate between mycotoxic and nontoxic isolates of A. flavus. The isolation of aflatoxigenic strains of A. flavus during post-harvest period of lobhiya seeds raise a serious concern over the quality of seeds and a threat to heath of consumers. It was concluded that Aspergillus flavus is responsible for postharvest spolilage of Lobhiya (Vigna unguiculata.

  19. Lumazine Peptides from the Marine-Derived Fungus Aspergillus terreus

    Directory of Open Access Journals (Sweden)

    Minjung You

    2015-03-01

    Full Text Available Terrelumamides A (1 and B (2, two new lumazine-containing peptides, were isolated from the culture broth of the marine-derived fungus Aspergillus terreus. From the results of combined spectroscopic and chemical analyses, the structures of these compounds were determined to be linear assemblies of 1-methyllumazine-6-carboxylic acid, an amino acid residue and anthranilic acid methyl ester connected by peptide bonds. These new compounds exhibited pharmacological activity by improving insulin sensitivity, which was evaluated in an adipogenesis model using human bone marrow mesenchymal stem cells. In addition, the compounds exhibited fluorescence changes upon binding to DNA, demonstrating their potential applications to DNA sequence recognition.

  20. Genetic relationships among strains of the Aspergillus niger aggregate

    DEFF Research Database (Denmark)

    Ferracin, L.M.; Frisvad, Jens Christian; Taniwaki, M.H.

    2009-01-01

    We analyzed the genetic relationships between 51 fungal isolates previously identified as A. niger aggregate, obtained from dried fruit samples from worldwide origin and 7 A. tubingensis obtained from Brazilian coffee beans samples. Greater fungal diversity was found in black sultanas. Aspergillus...... an association between extrolite patterns and molecular clustering is speculated. A. tubingensis were the second most frequent species and this species were clearly subdivided into two groups. The finding of two groups for A. tubingensis strains could not yet explain the contradictions found in the literature...

  1. Sequence analysis and overexpression of a pectin lyase gene (pel1) from Aspergillus oryzae KBN616.

    Science.gov (United States)

    Kitamoto, N; Yoshino-Yasuda, S; Ohmiya, K; Tsukagoshi, N

    2001-01-01

    A gene (pel1) encoding pectin lyase (Pel1) was isolated from a shoyu koji mold, Aspergillus oryzae KBN616, and characterized. The structural gene comprised 1,196 bp with a single intron. The ORF encoded 381 amino acids with a signal peptide of 20 amino acids. The deduced amino acid sequence showed high similarity to those of Aspergillus niger pectin lyases and Glomerella cingulata PnlA. The pel1 gene was successfully overexpressed under the promoter of the A. oryzae TEF1 gene. The molecular mass of the recombinant pectin lyase substantially coincided with that calculated based on nucleotide sequence.

  2. Aflatoxin production by Aspergillus Flavus on irradiated and non-irradiated stored egyptian bread

    International Nuclear Information System (INIS)

    Elbazza, Z.E.; Mahmoud, M.I.; Fahd, M.E.; Abu Seif, F.A.

    1986-01-01

    Aflatoxins production by Aspergillus Flavus isolate on several types of egyptian bread samples through a period of 45 days at different temperatures reveled that there was detectable fungal growth but no aflatoxins were produced at 5±1 degree or 37 degree. At 25 degree or 30 degree aflatoxin were produced after 5-6 days on the different bread types. Gamma irradiation of kaiser bread, previously inoculated with Aspergillus Flavus, reduced the amount of aflatoxins produced on subsequent storage for 8 weeks at 26±1 degree. The 5 KGy dose eliminated fungal growth and aflatoxin production. 4 tab

  3. Molecular detection and identification of Candida and Aspergillus spp. from clinical samples using real-time PCR.

    Science.gov (United States)

    Klingspor, L; Jalal, S

    2006-08-01

    This report describes the development of a real-time LightCycler assay for the detection and identification of Candida and Aspergillus spp., using the MagNa Pure LC Instrument for automated extraction of fungal DNA. The assay takes 5-6 h to perform. The oligonucleotide primers and probes used for species identification were derived from the DNA sequences of the 18S rRNA genes of various fungal pathogens. All samples were screened for Aspergillus and Candida to the genus level in the real-time PCR assay. If a sample was Candida-positive, typing to species level was performed using five species-specific probes. The assay detected and identified most of the clinically relevant Aspergillus and Candida spp. with a sensitivity of 2 CFU/mL blood. Amplification was 100% specific for all Aspergillus and Candida spp. tested. To assess clinical applicability, 1,650 consecutive samples (1,330 blood samples, 295 samples from other body fluids and 25 biopsy samples) from patients with suspected invasive fungal infections were analysed. In total, 114 (6.9%) samples were PCR-positive, 5.3% for Candida and 1.7% for Aspergillus spp. In patients with positive PCR results for Candida and Aspergillus, verification with conventional methods was possible in 83% and 50% of cases, respectively. In conclusion, the real-time PCR assay allows sensitive and specific detection and identification of fungal pathogens in vitro and in vivo.

  4. In vitro antifungal susceptibility of clinical species belonging to Aspergillus genus and Rhizopus oryzae.

    Science.gov (United States)

    Kachuei, R; Khodavaisy, S; Rezaie, S; Sharifynia, S

    2016-03-01

    Among filamentous fungal pathogens, Aspergillus spp. and zygomycetes account for highest rates of morbidity and mortality among immunocompromised patients. Recently developed antifungal drugs offer the potential to improve management and therapeutic outcomes of fungal infections. The aim of this study was to analyse the in vitro activities of voriconazole, itraconazole, amphotericin B and caspofungin against clinical isolates of Aspergillus spp. and Rhizopus oryzae. The in vitro antifungal susceptibility of 54 isolates belonging to different clinical isolates of Aspergillus spp. and R. oryzae was tested for four antifungal agents using a microdilution reference method (CLSI, M38-A2). All isolates were identified by typical colony and microscopic characteristics, and also characterized by molecular methods. Caspofungin (MEC range: 0.008-0.25 and MEC50: 0.0023μg/mL) was the most active drug in vitro against Aspergillus spp., followed by voriconazole (MIC range: 0.031-8 and MIC50: 0.5μg/mL), itraconazole (MIC range: 0.031-16 and MIC50: 0.25μg/mL), and amphotericin B (MIC range: 0.125-4 and MIC50: 0.5μg/mL), in order of decreasing activity. The caspofungin, voriconazole, and itraconazole demonstrated poor in vitro activity against R. oryzae isolates evaluated, followed by amphotericin B. This study demonstrates that caspofungin had good antifungal activity and azole agents had better activity than amphotericin B against Aspergillus species. Although, azole drugs are considered ineffective against R. oryzae. This result is just from a small scale in vitro susceptibility study and we did not take other factors into consideration. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  5. Equilibrium and kinetic studies of copper biosorption by dead Ceriporia lacerata biomass isolated from the litter of an invasive plant in China.

    Science.gov (United States)

    Li, Xiaona; Li, Airong; Long, Mingzhong; Tian, Xingjun

    2015-01-01

    Ceriporia lacerata, a strain of white-rot fungus isolated from the litter of an invasive plant (Solidago canadensis) in China, was little known about its properties and utilization. In this work, the copper(II) biosorption characteristics of formaldehyde inactivated C. lacerata biomass were examined as a function of initial pH, initial copper(II) concentration and contact time, and the adsorptive equilibrium and kinetics were simulated, too. The optimum pH was found to be 6.0 at experimental conditions of initial copper(II) concentration 100 mg/L, biomass dose 2 g/L, contact time 12 h, shaking rate 150 r/min and temperature 25°C. Biosorption equilibrium cost about 1 hour at experimental conditions of pH 6.0, initial copper(II) concentration 100 mg/L, C. lacerata dose 2 g/L, shaking rate 150 r/min and temperature 25°C. At optimum pH 6.0, highest copper(II) biosorption amounts were 6.79 and 7.76 mg/g for initial copper(II) concentration of 100 and 200 mg/L, respectively (with other experimental parameters of C. lacerata dose 2 g/L, shaking rate 150 r/min and temperature 25°C). The pseudo second-order adsorptive model gave the best adjustment for copper(II) biosorption kinetics. The equilibrium data fitted very well to both Langmuir and Freundlich adsorptive isotherm models. Without further acid or alkali treatment for improving adsorption properties, formaldehyde inactivated C. lacerata biomass possesses good biosorption characteristics on copper(II) removal from aqueous solutions.

  6. Inhibition of Growth of Fungi Isolated From Deteriorating Melon ...

    African Journals Online (AJOL)

    A study was conducted to investigate the effect of extracts of Punica granatum and Cymbopogon citratus on Aspergillus nivale, Rhizopus stolonifer , Mucor mucido and Aspergillus fumigatus isolated from deteriorating melon seed using radial growth technique. Phytochemical screening revealed that extracts the plants ...

  7. Inhibition of Growth of Fungi Isolated From Deteriorating Melon ...

    African Journals Online (AJOL)

    BSN

    Abstract. A study was conducted to investigate the effect of extracts of Punica granatum and. Cymbopogon citratus on Aspergillus nivale, Rhizopus stolonifer , Mucor mucido and. Aspergillus fumigatus isolated from deteriorating melon seed using radial growth technique. Phytochemical screening revealed that extracts the ...

  8. Lysine aminopeptidase of Aspergillus niger

    OpenAIRE

    Basten, D.E.J.W.; Visser, J.; Schaap, P.J.

    2001-01-01

    Conserved regions within the M1 family of metallo-aminopeptidases have been used to clone a zinc aminopeptidase from the industrially used fungus Aspergillus niger. The derived amino acid sequence of ApsA is highly similar to two yeast zinc aminopeptidases, LAPI and AAPI (53.3 and 50.9␘verall similarity, respectively), two members of the M1 family of metallo-aminopeptidases. The encoding gene was successfully overexpressed in A. niger and the overexpressed product was purified and characteriz...

  9. Determining the analytical specificity of PCR-based assays for the diagnosis of IA: What is Aspergillus?

    Science.gov (United States)

    Morton, C Oliver; White, P Lewis; Barnes, Rosemary A; Klingspor, Lena; Cuenca-Estrella, Manuel; Lagrou, Katrien; Bretagne, Stéphane; Melchers, Willem; Mengoli, Carlo; Caliendo, Angela M; Cogliati, Massimo; Debets-Ossenkopp, Yvette; Gorton, Rebecca; Hagen, Ferry; Halliday, Catriona; Hamal, Petr; Harvey-Wood, Kathleen; Jaton, Katia; Johnson, Gemma; Kidd, Sarah; Lengerova, Martina; Lass-Florl, Cornelia; Linton, Chris; Millon, Laurence; Morrissey, C Orla; Paholcsek, Melinda; Talento, Alida Fe; Ruhnke, Markus; Willinger, Birgit; Donnelly, J Peter; Loeffler, Juergen

    2017-06-01

    A wide array of PCR tests has been developed to aid the diagnosis of invasive aspergillosis (IA), providing technical diversity but limiting standardisation and acceptance. Methodological recommendations for testing blood samples using PCR exist, based on achieving optimal assay sensitivity to help exclude IA. Conversely, when testing more invasive samples (BAL, biopsy, CSF) emphasis is placed on confirming disease, so analytical specificity is paramount. This multicenter study examined the analytical specificity of PCR methods for detecting IA by blind testing a panel of DNA extracted from a various fungal species to explore the range of Aspergillus species that could be detected, but also potential cross reactivity with other fungal species. Positivity rates were calculated and regression analysis was performed to determine any associations between technical specifications and performance. The accuracy of Aspergillus genus specific assays was 71.8%, significantly greater (P < .0001) than assays specific for individual Aspergillus species (47.2%). For genus specific assays the most often missed species were A. lentulus (25.0%), A. versicolor (24.1%), A. terreus (16.1%), A. flavus (15.2%), A. niger (13.4%), and A. fumigatus (6.2%). There was a significant positive association between accuracy and using an Aspergillus genus PCR assay targeting the rRNA genes (P = .0011). Conversely, there was a significant association between rRNA PCR targets and false positivity (P = .0032). To conclude current Aspergillus PCR assays are better suited for detecting A. fumigatus, with inferior detection of most other Aspergillus species. The use of an Aspergillus genus specific PCR assay targeting the rRNA genes is preferential. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  10. Heterologous expression of Gaeumannomyces graminis lipoxygenase in Aspergillus nidulans

    NARCIS (Netherlands)

    Heshof, R.; Schayck, van J.P.; Tamayo Ramos, J.A.; Graaff, de L.H.

    2014-01-01

    Aspergillus sp. contain ppo genes coding for Ppo enzymes that produce oxylipins from polyunsaturated fatty acids. These oxylipins function as signal molecules in sporulation and influence the asexual to sexual ratio of Aspergillus sp. Fungi like Aspergillus nidulans and Aspergillus niger contain

  11. Occurrence of Aflatoxins and Aflatoxin-Producing Strains of Aspergillus spp. in Soybeans 1

    Science.gov (United States)

    Bean, George A.; Schillinger, John A.; Klarman, William L.

    1972-01-01

    Above average rainfall in Maryland during August, September, and October 1971 resulted in heavy mold growth in soybeans while still in the field. Of 28 samples of soybean seed, aflatoxins were found in 14, 2 of which had been used in poultry feed. Aflatoxins were identified by thin-layer chromatography, spectrophotometry, and chicken embryo bioassay. Aspergillus spp. were isolated from 11 samples, and 5 of these isolates produced aflatoxins when grown in liquid culture. PMID:4673021

  12. Fumonisin B2 production by Aspergillus niger in Thai coffee beans

    DEFF Research Database (Denmark)

    Noonim, P.; Mahakarnchanaku, W.; Nielsen, Kristian Fog

    2009-01-01

    by black Aspergilli. No Fusarium species known to produce fumonisin were detected, but black Aspergilli had high incidences on both Arabica and Robusta Thai coffee beans. Liquid chromatography (LC) with high-resolution mass spectrometric (HRMS) detection showed that 67% of Aspergillus niger isolates from....... This is the first record of freshly isolated A. niger strains producing fumonisins and the first report on the natural occurrence of FB2 and FB4 in coffee....

  13. Diversity of Aspergillus section Nigri on the surface of Vitis labrusca and its hybrid grapes

    DEFF Research Database (Denmark)

    Ferranti, Larissa de Souza; Fungaro, Maria Helena P; Massi, Fernanda Pelisson

    2018-01-01

    This study investigated the presence of Aspergillus species belonging to Aspergillus section Nigri on Vitis labrusca and its hybrid grapes grown in Brazil. The ability of the fungi isolates to produce ochratoxin A (OTA) and fumonisin B2 (FB2) as well as the presence of these mycotoxins in the gra......This study investigated the presence of Aspergillus species belonging to Aspergillus section Nigri on Vitis labrusca and its hybrid grapes grown in Brazil. The ability of the fungi isolates to produce ochratoxin A (OTA) and fumonisin B2 (FB2) as well as the presence of these mycotoxins...... in the grapes were also studied. Eighty-eight samples were collected from the main grape producing states in Brazil: Rio Grande do Sul (n=30), Pernambuco (n=21), São Paulo (n=21) and Paraná (n=16). The highest average contamination level by A. section Nigri occurred on the grapes from Pernambuco (66...... and A. vadensis. Regarding mycotoxin-production capacity, 3.2% of the total A. section Nigri isolates (2042) were positive for OTA production and from A. niger "aggregate" (373) tested, 42.1% were FB2 producers. However, none of the 88 grape samples were contaminated with these mycotoxins....

  14. Ribonuclease Production by Aspergillus species

    Directory of Open Access Journals (Sweden)

    Gomes Eleni

    1998-01-01

    Full Text Available Ribonuclease production by Aspergillus flavipes, A. sulphureus and A. fischeri in semi-synthetic medium, after 24-144 hours at 30ºC under shaking, was studied. After cultivation, the medium was separated from micelia by filtration and the resultant solution was used as enzymatic extract. The highest amount of biomass and RNase was obtained after 96 hours of cultivation. The enzymes produced by three species presented similar characteristics, with optimum temperature at 55ºC and two peaks of activity at pH 4.5 and 7.0. A. flavipes RNases were more sensitive to temperature: 50% of the initial activity was lost after 1 hour at 70ºC. After this heat treatment, RNase of A. sulphureus lost 30% of this activity and that of A. fischeri only 16%. The nucleotides released by enzimatic hydrolysis of RNA were separated by ion exchange chromatography in a AG-1X8-formiate column and identified by paper chromatography. This procedure indicated that the raw enzymatic extract of Aspergillus flavipes is able to hydrolyze RNA, releasing 3'-nucleotides monophosphate at pH 4.5 and 3' and 5'-nucleotides monophosphate at pH 7.0 and 8.5. This result suggests that this strain produces two different types of RNase, one acidic and other alcaline, with different specificities.

  15. Activity of echinocandins and triazoles against a contemporary (2012) worldwide collection of yeast and moulds collected from invasive infections.

    Science.gov (United States)

    Castanheira, Mariana; Messer, Shawn A; Jones, Ronald N; Farrell, David J; Pfaller, Michael A

    2014-10-01

    In this study, 1717 fungal clinical isolates causing invasive fungal infections were evaluated against nine antifungal agents using Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution methods. The isolates comprised 1487 Candida spp., 109 Aspergillus spp., 86 non-Candida yeasts (including 52 isolates of Cryptococcus neoformans) and 35 rare moulds obtained during 2012 from 72 hospitals worldwide. Echinocandin resistance among Candida spp. was low, and resistance rates to anidulafungin, caspofungin and micafungin varied from 0.0% to 2.8% among different species. Echinocandin-resistant Candida glabrata were shown to have fks mutations (fks2 HS1 F659Y, F659del, S663F and S663P), and fluconazole resistance was also observed in those strains. One Candida krusei and one Candida dubliniensis had L701M or S645P fks1 mutations, respectively. Candida tropicalis and C. glabrata had higher fluconazole resistance rates of 6.1% and 6.9%, respectively, compared with other Candida spp. Fluconazole-resistant C. tropicalis were collected in five countries (USA, China, Germany, Belgium and Thailand). Voriconazole was active against all Candida spp., inhibiting 91.2-99.7% of isolates using species-specific breakpoints. All agents except for the echinocandins and posaconazole were active against Cr. neoformans. Triazoles were active against other yeasts [MIC90 (minimum inhibitory concentration encompassing 90% of isolates tested), 2μg/mL]. The echinocandins and the mould-active triazoles were active against Aspergillus [MIC/MEC90 (minimum effective concentration encompassing 90% of isolates tested) range, 0.015-2μg/mL], but the activity of these agents was limited against uncommon mould species (MIC/MEC90 range, 4μg/mL to >16μg/mL). Copyright © 2014 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  16. Rhino-oculo-cerebral aspergillus and mucor co-infections in an immunocompromised patient with type 2 diabetes mellitus

    Directory of Open Access Journals (Sweden)

    Kalidas Rit

    2014-01-01

    Full Text Available Mucormycosis are pathogenic moulds of the mucorales species usually occurring in immunocompromised patients or in patients with uncontrolled diabetes mellitus. Aspergillosis is the clinical condition caused by Aspergillus species and may cause an invasive disease with high case fatality rate, especially in immunosuppressed patients. A 46-year-old male patient with Type 2 diabetes mellitus with underlying malignancy presented with proptosis of left eye. Combined infections of Mucor and Aspergillus were diagnosed by means of computed tomography (CT scan and biopsy. Treatment with Amphotericin B and Voriconazole was started, the patient died within 3 months, from multi-organ failure.

  17. The biodiversity of Aspergillus section Flavi in brazil nuts: from rainforest to consumer.

    Science.gov (United States)

    Calderari, Thaiane O; Iamanaka, Beatriz T; Frisvad, Jens C; Pitt, John I; Sartori, Daniele; Pereira, Jose Luiz; Fungaro, Maria Helena P; Taniwaki, Marta H

    2013-01-01

    A total of 288 brazil nut samples (173 kernel and 115 shell) from the Amazon rainforest region and São Paulo State, Brazil were collected at different stages of brazil nut production. Samples were analysed for: percentages of aflatoxigenic fungal species and potential for aflatoxin production and presence of aflatoxins. Aspergillus nomius was the most common species found (1235 isolates) which amounted to 30% of the total species with potential to produce aflatoxins. This species is of concern since 100% of all isolates produced aflatoxins B(1), B(2), G(1) and G(2). Aspergillus flavus was almost equally common (1212 isolates) although only 46% produced aflatoxins under laboratory conditions, and only aflatoxins B(1) and B(2). Low number of other species with the potential to produce aflatoxins was isolated: Aspergillus arachidicola and Aspergillus bombycis produced B and G aflatoxins whilst Aspergillus pseudotamarii produced only aflatoxin B(1). The total aflatoxin levels found in samples taken from the rainforests was 0.7 μg/kg, from processing plants before and after sorting 8.0 and 0.1 μg/kg respectively, from street markets in the Amazon region 6.3 μg/kg and from supermarkets in São Paulo State 0.2 μg/kg. Processing, which included manual or mechanical sorting and drying at 60°C for 30 to 36 h, eliminated on average more than 98% of total aflatoxins. These results showed that sorting is a very effective way to decrease aflatoxin content in brazil nuts. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. Manifestation of invasive pulmonary aspergillosis in the formation of mycetoma

    Energy Technology Data Exchange (ETDEWEB)

    Roos, N.; Peters, P.E.; Schellong, S.; Eiff, M. von

    1989-05-01

    Invasive pulmonary aspergillosis is characterized by radiological signs allowing a correct diagnosis, including differentiation from pulmonary candidiasis, when they are associated with appropriate clinical symptoms (neutropenia and fever persisting despite broad-spectrum antibiotics). In particular the formation of a pulmonary mycetoma in a previously normal lung is one of these signs. Unlike a simple fungus ball (the saprophytic form of aspergillosis), the rounded density of invasive pulmonary aspergillosis consists of sequestrum of devitalized lung tissue owing to blood vessel invasion by Aspergillus hyphae. This morphologic phenomenon is demonstrated in the present case report and is discussed together with the other roentgenological signs of the invasive aspergillosis.

  19. Manifestation of invasive pulmonary aspergillosis in the formation of mycetoma

    International Nuclear Information System (INIS)

    Roos, N.; Peters, P.E.; Schellong, S.; Eiff, M. von

    1989-01-01

    Invasive pulmonary aspergillosis is characterized by radiological signs allowing a correct diagnosis, including differentiation from pulmonary candidiasis, when they are associated with appropriate clinical symptoms (neutropenia and fever persisting despite broad-spectrum antibiotics). In particular the formation of a pulmonary mycetoma in a previously normal lung is one of these signs. Unlike a simple fungus ball (the saprophytic form of aspergillosis), the rounded density of invasive pulmonary aspergillosis consists of sequestrum of devitalized lung tissue owing to blood vessel invasion by Aspergillus hyphae. This morphologic phenomenon is demonstrated in the present case report and is discussed together with the other roentgenological signs of the invasive aspergillosis. (orig.) [de

  20. Production and Preliminary Characterization of Alkaline Protease from Aspergillus flavus and Aspergillus terreus

    OpenAIRE

    Chellapandi, P.

    2010-01-01

    Proteases are being an industrial candidate, which are widely used in food, bakery, and beverage and detergent industry. In leather industry, alkaline proteases are exhibiting a prominent role in unhairing and bating processes. An extensive use of filamentous fungi, especially Aspergillus species has been studied elaborately. Although, the significant application of alkaline protease produced from these strains in leather industry is being limited. Aspergillus flavus and Aspergillus terreus f...

  1. Targeting zinc homeostasis to combat Aspergillus fumigatus infections

    Directory of Open Access Journals (Sweden)

    Rocío eVicentefranqueira

    2015-02-01

    Full Text Available Aspergillus fumigatus is able to invade and grow in the lungs of immunosuppressed individuals and causes invasive pulmonary aspergillosis. The concentration of free zinc in living tissues is much lower than that required for optimal fungal growth in vitro because most of it is tightly bound to proteins. To obtain efficiently zinc from a living host A. fumigatus uses the zinc transporters ZrfA, ZrfB and ZrfC. The ZafA transcriptional regulator induces the expression of all these transporters and is essential for virulence. Thus, ZafA could be targeted therapeutically to inhibit fungal growth. The ZrfC transporter plays the major role in zinc acquisition from the host whereas ZrfA and ZrfB rather have a supplementary role to that of ZrfC. In addition, only ZrfC enables A. fumigatus to overcome the inhibitory effect of calprotectin, which is an antimicrobial Zn/Mn-chelating protein synthesized and released by neutrophils within the fungal abscesses of immunosuppressed non-leucopenic animals. Hence, fungal survival in these animals would be undermined upon blocking therapeutically the function of ZrfC. Therefore, both ZafA and ZrfC have emerged as promising targets for the discovery of new antifungals to treat Aspergillus infections.

  2. Chronic obstructive pulmonary disease patients with invasive pulmonary aspergillosis: benefits of intensive care?

    Science.gov (United States)

    Bulpa, P A; Dive, A M; Garrino, M G; Delos, M A; Gonzalez, M R; Evrard, P A; Glupczynski, Y; Installé, E J

    2001-01-01

    Invasive pulmonary aspergillosis (IPA) is increasingly recognized as a cause of acute respiratory failure in patients with chronic obstructive pulmonary disease (COPD) treated with corticosteroids. For these patients admission in intensive care unit (ICU) is often required for life-support and mechanical ventilation. Whether this approach improves outcome is unknown. Retrospective study in a university hospital intensive care unit. Between November 1993 and December 1997, 23 COPD patients were admitted in our ICU and received antifungal agents for possible IPA. None. The clinical features and the outcome were reviewed. Diagnosis of IPA was classified as confirmed (positive lung tissue biopsy and/or autopsy) or probable (repeated isolation of Aspergillus from the airways with consistent clinical and radiological findings). Among the 23 patients treated for Aspergillus, 16 fulfilling these criteria for IPA were studied. Steroids had been administered at home to all patients but one and were increased during hospitalization in all. Twelve patients suffered a worsening of their bronchospasm precipitating acute respiratory failure. During ICU stay all patients required mechanical ventilation for acute respiratory failure. Although amphotericin B deoxycholate was started when IPA was suspected (0.5-1.5 mg/kg per day), all patients died in septic shock (n = 5) or in multiple-organ failure. The poor prognosis of intubated COPD patients with IPA, in spite of antifungal treatment suggests that further studies are required to define the limits and indications for ICU management of these patients.

  3. Cepas invasivas de pneumococo isoladas de crianças e adolescentes em Salvador Invasive pneumococcal strains isolated from children and adolescents in Salvador

    Directory of Open Access Journals (Sweden)

    Cristiana M. Nascimento-Carvalho

    2003-06-01

    Full Text Available OBJETIVOS: descrever resistência antimicrobiana e sorotipos de cepas de pneumococo. MÉTODOS: durante 57 meses, foi conduzida uma vigilância de cepas invasivas de pneumococo de pacientes com idade OBJECTIVE: describe the antimicrobial resistance and serotype distribution of pneumococcal strains. METHODS: in a 57-month period, a laboratory-based surveillance of invasive pneumococcal strains from patients aged < 20 years was conducted. Pneumococcus was identified by means of tests for solubility in bile and optochin. Pneumococcal resistance to penicillin was screened by 1µg oxacillin disc and minimal inhibitory concentration was determined for the strains not susceptible to penicillin. Disc diffusion and broth microdilution methods were used for surveillance of resistance to other antimicrobials. Pneumococci were serotyped by means of the Neufeld-Quellung reactions. RESULTS: of 70 patients, 57.1% were males. The mean age was 1.92 yrs (mean 3.19 + 3.66 yrs, range 1 month to 19.5 yrs; 52.9% and 81.4% were < 2 yrs and < 5 yrs, respectively. The strains were isolated from blood (91.4%, CSF (2.9%, pleural (2.9%, peritoneal (1.4% and abscess (1.4% fluids from patients with pneumonia (77.1%, fever without localizing signs (10.0%, meningitis (4.3%, others (8.6%. Resistance was detected to penicillin (20.0%, trimethoprim-sulfamethoxazole (65.7%, tetracycline (21.4%, ofloxacin (6.3%, erythromycin (5.7%, clindamycin (2.9%. All tested strains were susceptible to chloramphenicol and vancomycin. Among penicillin-resistant strains, high resistance was detected in one, the same that showed intermediate resistance to cefotaxime. The most frequent serotypes were: 14 (22.9%, 5 and 6A (10.0% each, 6B and 19F (8.6% each, 9V, 18C and 23F (5.7% each. Resistance to penicillin was detected in serotypes 14 (71.4%, 6B and 19F (14.3% each. CONCLUSIONS: of 70 strains, 67.2% were classified as serotypes included in the heptavalent conjugate pneumococcal vaccine as well as

  4. Two new aflatoxin producing species, and an overview of Aspergillus section Flavi

    DEFF Research Database (Denmark)

    Varga, J.; Frisvad, Jens Christian; Samson, R. A.

    2011-01-01

    Aspergillus subgenus Circumdati section Flavi includes species with usually biseriate conidial heads, in shades of yellow-green to brown, and dark sclerotia. Several species assigned to this section are either important mycotoxin producers including aflatoxins, cyclopiazonic acid, ochratoxins...... and extrolite profiles. Aspergillus pseudocaelatus is represented by a single isolate collected from Arachis burkartii leaf in Argentina, is closely related to the non-aflatoxin producing A. caelatus, and produces aflatoxins B & G, cyclopiazonic acid and kojic acid, while A. pseudonomius was isolated from...... insects and soil in the USA. This species is related to A. nomius, and produces aflatoxin B-1 (but not G-type aflatoxins), chrysogine and kojic acid. In order to prove the aflatoxin producing abilities of the isolates, phylogenetic analysis of three genes taking part in aflatoxin biosynthesis, including...

  5. Endolithic fungi from deep sea calcareous substrata: isolation and laboratory studies

    Digital Repository Service at National Institute of Oceanography (India)

    Raghukumar, C.; Raghukumar, S.; Sharma, S.; Chandramohan, D.

    Fungi were isolated from calcareous animal shells at 100-860 m depth in the Bay of Bengal. All the species isolated were terrestrial ones, Aspergillus restrictus Smith being the most common. The behaviour of this fungus at various hydrostatic...

  6. A prospective international Aspergillus terreus survey

    DEFF Research Database (Denmark)

    Risslegger, B; Zoran, T; Lackner, M

    2017-01-01

    OBJECTIVES: A prospective international multicentre surveillance study was conducted to investigate the prevalence and amphotericin B susceptibility of Aspergillus terreus species complex infections. METHODS: A total of 370 cases from 21 countries were evaluated. RESULTS: The overall prevalence o...

  7. Aspergillus arthritis: analysis of clinical manifestations, diagnosis, and treatment of 31 reported cases.

    Science.gov (United States)

    Gamaletsou, Maria N; Rammaert, Blandine; Bueno, Marimelle A; Sipsas, Nikolaos V; Moriyama, Brad; Kontoyiannis, Dimitrios P; Roilides, Emmanuel; Zeller, Valerie; Taj-Aldeen, Saad J; Henry, Michael; Petraitis, Vidmantas; Denning, David W; Lortholary, Olivier; Walsh, Thomas J

    2017-04-01

    Aspergillus arthritis is a debilitating form of invasive aspergillosis. Little is known about its epidemiology, clinical manifestations, laboratory features, treatment, and prognosis. Cases of Aspergillus arthritis were reviewed in the English literature from 1967 through 2015 for variables of arthritis with Aspergillus spp. recovered from joint and/or adjacent bone, underlying conditions, symptoms, signs, inflammatory biomarkers, diagnostic imaging, management, and outcome. Among 31 evaluable cases, 87% were males and 13% pediatric. Median age was 50 y (range 1-83 y). Seventeen (55%) patients were immunosuppressed with such conditions as hematological malignancies (26%), corticosteroids (39%), and/or transplantation (26%). Approximately one-half (52%) of patients had hematogenous seeding of the joint, and more than 80% had de novo infection with no prior antifungal therapy. Oligoarticular infection (2-3 joints) occurred in 45% and contiguous osteomyelitis was present in 61%. Clinical manifestations included pain (87%), edema (26%), and limited function (23%), with knees (35%), intervertebral discs (26%), and hips (16%) being most commonly infected. Aspergillus fumigatus constituted 77% of cases followed by Aspergillus flavus in 13%, Aspergillus niger in 3%, and not specified in 7%. Median ESR was 90 mm/hr and median CRP was 3.6 mg/dl. Median synovial fluid WBC was 17,200/μL (7,300-128,000) with 72% PMNs (range 61-92). Osteolysis occurred in 35%, and soft-tissue extension 47%. Nineteen patients (61%) were managed with combined medical and surgical therapy, 10 (32%) with medical therapy only, and 2 (6%) surgery only. Amphotericin B and itraconazole were the most frequently used agents with median duration of therapy of 219 days (range 30-545). Surgical interventions included debridement in 61%, drainage 19%, and amputation 6%. Complete or partial response was achieved in 71% and relapse occurred in 16%. Medical therapy was reinstituted with successful outcome in

  8. Black Aspergillus species as ochratoxin A producers in Portuguese wine grapes.

    Science.gov (United States)

    Serra, Rita; Abrunhosa, Luís; Kozakiewicz, Zofia; Venâncio, Armando

    2003-11-15

    To evaluate the incidence of fungi producing ochratoxin A (OA) in Portuguese wine grapes, a survey was conducted in 11 vineyards, from four winemaking regions each with distinct climatic conditions. From setting to the harvesting period, a total of 1,650 berries were sampled by plating methods. Out of 370 aspergilli and 301 Penicillium strains isolated, 14% of the aspergilli were OA-producing strains. None of the penicillia were OA-producing strains. The black aspergilli were predominant (90%). All Aspergillus strains were tested in vitro for OA production and all were preserved in the Micoteca da Universidade do Minho (MUM) culture collection. Most of the Aspergillus carbonarius (97%) and 4% of the Aspergillus niger aggregate strains were OA producers. Almost all ochratoxigenic strains were isolated at harvest time, mainly in the regions with a Mediterranean climate. In the vineyards sampled, the percentage of colonized berries with ochratoxigenic strains was up to 38%. The vineyards from the region with Atlantic influences, with high rainfall, exhibited the lowest occurrence of Aspergillus and ochratoxigenic strains, 0% to 10% and 0% to 2% colonized berries, respectively. Data obtained here supports the hypothesis that A. carbonarius and occasionally A. niger, are the main producers of OA in grapes. In this study, the highest incidence of these fungi occurred in vineyards with a Mediterranean climate.

  9. Polyphasic taxonomy of Aspergillus fumigatus and related species

    DEFF Research Database (Denmark)

    Hong, S.B.; Go, S.J.; Shin, H.D.

    2005-01-01

    . fumigatus sensu stricto species. A. lentulus including isolates from clinical origin, Korean soil and from a dolphin Clustered into an isolated group based on beta-tubulin, calmodulin and actin gene sequences, differing from A. fumigalus by morphological characters, growth temperature and extrolite profile......The variability within Aspergillus fumigalus Fresenius and related species was examined using macro-, micro-morphology, growth temperature regimes and extrolite patterns. In addition, DNA analyses including partial beta-tubulin, calmodulin and actin gene sequences were used. Detailed examination...... of strains, considered as A. fumigatus earlier, showed that they could be divided into four groups including A. fumigatus sensu stricto, A. lentulus and two new species. The intraspecific genetic variability within A. fumigatus sensu stricto was low, the sequence differences among 23 strains of the species...

  10. Biodegradation of alpha and beta endosulfan by Aspergillus sydoni.

    Science.gov (United States)

    Goswami, Supriya; Vig, Komal; Singh, Dileep K

    2009-05-01

    The biodegradation of endosulfan and the metabolites formed were studied using fungi both in broth culture as well as in soil microcosm. Fungal strains were isolated from soil and grown in broth Czapek-dox medium. The strain which utilized endosulfan and showed maximum growth was selected for detailed studies. Maximum degrading capability in shake flask culture was shown by Aspergillus sydoni which degraded 95% of endosulfan alpha and 97% of endosulfan beta in 18 d of incubation. Soil microcosm study was also carried out using this strain in six different treatments. Endosulfan sulfate was the main metabolite formed along with small quantity of endosulfan ether and endosulfan lactone both in broth culture and soil microcosm. This isolated fungal strain will be a potential source for endosulfan degrading enzymes and can be used for bioremediation at the contaminated sites.

  11. Aspergillus sect. Aeni sect. nov., a new section of the genus for A.karnatakaensis sp. nov. and some allied fungi.

    Science.gov (United States)

    Varga, János; Frisvad, Jens C; Samson, Robert A

    2010-12-01

    The new species Aspergilluskarnatakaensis sp. nov. is described and illustrated. All three isolates of this species were isolated from Indian soil; two from soil under a coconut palm in a coffee plantation in Karnataka, and one from soil in the Machrar river bed in Bansa district. This species is closely related to, but clearly distinct, from A. aeneus based on β-tubulin or calmodulin sequence data. Sequences of the ITS region of these two species are identical. Aspergillus karnatakaensis produced terrein, gregatins, asteltoxin, karnatakafurans A and B and the unknown metabolite, provisionally named NIDU. Aspergillus karnatakaensis belongs to a well-defined clade within Aspergillus subgenus Nidulantes together with eight other species including A. aeneus, A. crustosus, A. eburneocremeus, A. heyangensis, and the teleomorph producing-species Emericella bicolor, E. discophora, E. spectabilis, and E. foeniculicola. This clade is placed in a new section, Aspergillus sect. Aenei sect. nov. All teleomorph species assigned to this section are able to produce sterigmatocystin.

  12. Invasive Species

    Science.gov (United States)

    Invasive species have significantly changed the Great Lakes ecosystem. An invasive species is a plant or animal that is not native to an ecosystem, and whose introduction is likely to cause economic, human health, or environmental damage.

  13. Post-genomic insights into the plant polysaccharide degradation potential of Aspergillus nidulans and comparison to Aspergillus niger and Aspergillus oryzae

    NARCIS (Netherlands)

    Coutinho, Pedro M; Andersen, Mikael R; Kolenova, Katarina; vanKuyk, Patricia A; Benoit, Isabelle; Gruben, Birgit S; Trejo-Aguilar, Blanca; Visser, Hans; van Solingen, Piet; Pakula, Tiina; Seiboth, Bernard; Battaglia, Evy; Aguilar-Osorio, Guillermo; de Jong, Jan F; Ohm, Robin A; Aguilar, Mariana; Henrissat, Bernard; Nielsen, Jens; Stålbrand, Henrik; de Vries, Ronald P

    The plant polysaccharide degradative potential of Aspergillus nidulans was analysed in detail and compared to that of Aspergillus niger and Aspergillus oryzae using a combination of bioinformatics, physiology and transcriptomics. Manual verification indicated that 28.4% of the A. nidulans ORFs

  14. Bacteria and fungi isolated from housefly ( Musca domestica L.) larvae

    African Journals Online (AJOL)

    A variety of microorganisms, which includes the pathogenic Staphylococcus aureus, Pseudomonas aerruginosa, Aspergillus tamarii and Bacullus cereus were found. Also, nonpathogenic microbes werer recovered including Bacillus subtilis and Stroplococcus faecalis. Keywords: Microorganism, isolation, housefly larvae.

  15. Isolation of Ascomycetous Fungi from a Tertiary Institution Campus ...

    African Journals Online (AJOL)

    The predominant Ascomycetous fungi isolated include among others; Aspergillus niger, Fusarium solani, Fusarium oxysporum, Penicillium italicum, Fusarium acuminatum, Fusarium culmorum, Candida albicans, Botrytis cinerea, Geotrichum candidum, Trichoderma viride, Verticillium lateritum, Curvularia palescens ...

  16. Aspergillus and Penicillium (Eurotiales: Trichocomaceae) in soils of the Brazilian tropical dry forest: diversity in an area of environmental preservation.

    Science.gov (United States)

    Barbosa, Renan do Nascimento; Bezerra, Jadson Diogo Pereira; Costa, Phelipe Manoel Oller; de Lima-Júnior, Nelson Correia; Alves de Souza Galvão, Ivana Roberta Gomes; Alves dos Santos-Júnior, Anthony; Fernandes, Maria José; de Souza-Motta, Cristina Maria; Oliveira, Neiva Tinti

    2016-03-01

    Soil is a complex biological system that plays a key role for plants and animals, especially in dry forests such as the Caatinga. Fungi from soils, such as Aspergillus and Penicillium, can be used as bioindica- tors for biodiversity conservation. The aim of this study was to isolate and identify species of Aspergillus and Penicillium in soil, from the municipalities of Tupanatinga and Ibimirim, with dry forests, in the Catimbau National Park. Five collections were performed in each area during the drought season of 2012, totaling 25 soil samples per area. Fungi were isolated by suspending soil samples in sterile distilled water and plating on Sabouraud Agar media plus Chloramphenicol and Rose Bengal, and Glycerol Dicloran Agar. Isolates were identified by morphological taxonomy in the Culture Collection Laboratory and confirmed by sequencing of the Internal Transcribed Spacer of rDNA. A total of 42 species were identified, of which 22 belong to the genus Aspergillus and 20 to Penicillium. Penicillium isolates showed uniform distribution from the collecting area in Tupanatinga, and the evenness indices found were 0.92 and 0.88 in Tupanatinga and Ibimirim, respectively. Among isolates of Aspergillus evenness, the value found in Tupanatinga (0.85) was very close to that found in Ibimirim (0.86). High diversity and low dominance of fungi in soil samples was observed. These results con- tributed to the estimation of fungal diversity in dry environments of the Caatinga, where diversity is decreasing in soils that have undergone disturbance.

  17. Potential Capsule Switching from Serogroup Y to B: The Characterization of Three such Neisseria meningitidis Isolates Causing Invasive Meningococcal Disease in Canada

    Directory of Open Access Journals (Sweden)

    Raymond SW Tsang

    2005-01-01

    Full Text Available Three group B Neisseria meningitidis isolates, recovered from meningococcal disease cases in Canada and typed as B:2c:P1.5, were characterized. Multilocus sequence typing showed that all three isolates were related because of an identical sequence type (ST 573. Isolates typed as 2c:P1.5 are common in serogroup Y meningococci but rare in isolates from serogroups B or C. Although no serogroup Y isolates have been typed as ST-573, eight isolates showed five to six housekeeping gene alleles that were identical to that of ST-573. This suggested that the B:2c:P1.5 isolates may have originated from serogroup Y organisms, possibly by capsule switching.

  18. Identification and toxigenic potential of the industrially important fungi, Aspergillus oryzae and Aspergillus sojae

    DEFF Research Database (Denmark)

    Jørgensen, Thomas R

    2007-01-01

    Mold strains belonging to the species Aspergillus oryzae and Aspergillus sojae are highly valued as koji molds in the traditional preparation of fermented foods, such as miso, sake, and shoyu, and as protein production hosts in modern industrial processes. A. oryzae and A. sojae are relatives...

  19. Variability in Galactomannan detection by platelia Aspergillus EIA™ according to the Aspergillus species

    Directory of Open Access Journals (Sweden)

    Melissa Orzechowski Xavier

    2013-06-01

    Full Text Available Here we investigate the extent to which different Aspergillus species release galactomannan (GM in vitro. Marked variability was observed in GM reactivity between and within Aspergillus species, with A. terreus strains showing the highest GM indexes. The in vivo significance of these findings remains to be determined.

  20. Invasive Pulmonary Aspergillosis in the Intensive Care Unit: A Case Report

    Directory of Open Access Journals (Sweden)

    Mehmet Akif Yazar

    2016-04-01

    Full Text Available SUMMARY Invasive pulmonary aspergillosis (IPA is an infection rarely seen in intensive care units (ICU. We aimed to discuss the case of IPA found in a patient followed up in our ICU due to chronic obstructive pulmonary disease (COPD. A 58-year old patient diagnosed with COPD has been admitted to our ICU due to respiratory failure. There were not any other diseases except from COPD. In his radiological imaging there were diffuse bilateral infiltrates in the chest X-ray and diffuse infiltration in the thorax tomography. Aspergillus spp was isolated in the bronchoalveolar lavage sample from patient and caspofungin was added to the treatment. On the 12th day of the treatment, it was observed that the results of the control radiological imaging became normal and there was no isolation in the control cultures. In recent years, fungal infections have been determined at an increasing rate in the patients who are not immunosuppressed and hospitalized in ICU. In the IPA cases, it has been shown that such underlying causes as diabetes, malnutrition, use of steroid, uremia, cirrhosis and COPD may create risk. Our patient did not have other disease except from COPD and there was a story of prolonged using of steroid. Steroids are quite frequently used in the ICUs. In the patients with chronic pulmonary diseases, even if it is low dose and short-term, the treatment of steroid is accepted as a risk factor in terms of IPA. Since signs and symptoms are atypical in this group of patients, it is rather difficult to diagnose IPA in the short term. The isolation of aspergillus in our patient’s tracheal aspirate taken earlier enabled us to start the treatment early. Although there are not comprehensive studies, it should be kept in mind that İPA may develop in the patients who is using long-term steroid, is admitted to the ICUs due to COPD exacerbations without any other disease.

  1. Characterization of Aspergillus species on Brazil nut from the Brazilian Amazonian region and development of a PCR assay for identification at the genus level.

    Science.gov (United States)

    Midorikawa, Glaucia E O; de Sousa, Maria de Lourdes M; Freitas Silva, Otniel; Dias, Jurema do Socorro A; Kanzaki, Luis I B; Hanada, Rogerio E; Mesquita, Renata M L C; Gonçalves, Rivadalve C; Alvares, Virginia S; Bittencourt, Daniela M C; Miller, Robert N G

    2014-05-30

    Brazil nut is a protein-rich extractivist tree crop in the Amazon region. Fungal contamination of shells and kernel material frequently includes the presence of aflatoxigenic Aspergillus species from the section Flavi. Aflatoxins are polyketide secondary metabolites, which are hepatotoxic carcinogens in mammals. The objectives of this study were to identify Aspergillus species occurring on Brazil nut grown in different states in the Brazilian Amazon region and develop a specific PCR method for collective identification of member species of the genus Aspergillus. Polyphasic identification of 137 Aspergillus strains isolated from Brazil nut shell material from cooperatives across the Brazilian Amazon states of Acre, Amapá and Amazonas revealed five species, with Aspergillus section Flavi species A. nomius and A. flavus the most abundant. PCR primers ASP_GEN_MTSSU_F1 and ASP_GEN_MTSSU_R1 were designed for the genus Aspergillus, targeting a portion of the mitochondrial small subunit ribosomal RNA gene. Primer specificity was validated through both electronic PCR against target gene sequences at Genbank and in PCR reactions against DNA from Aspergillus species and other fungal genera common on Brazil nut. Collective differentiation of the observed section Flavi species A. flavus, A. nomius and A. tamarii from other Aspergillus species was possible on the basis of RFLP polymorphism. Given the abundance of Aspergillus section Flavi species A. nomius and A. flavus observed on Brazil nut, and associated risk of mycotoxin accumulation, simple identification methods for such mycotoxigenic species are of importance for Hazard Analysis Critical Control Point system implementation. The assay for the genus Aspergillus represents progress towards specific PCR identification and detection of mycotoxigenic species.

  2. Preliminary studies of the biosynthesis of Austin. [Aspergillus ustus

    Energy Technology Data Exchange (ETDEWEB)

    Wicnienski, N.A.

    1979-01-01

    Aspergillus ustus is one of the most prevalent fungi in the soil. There are now two reports of the occurrence of toxin-producing strains of this fungus on stored foodstuffs. In addition, strains of A. ustus have been isolated along with Penicillium species from samples of South African cheeses. All A. ustus isolates tested were judged to be highly toxic to ducklings when grown on maize meal, however, the toxins involved were not isolated. Austin is the trivial name of one of the toxins made by the fungus found on stored food. Preliminary work to studying the biosynthesis of this compound using /sup 13/C-labeled sodium acetate is reported here. The feasibility of the biosynthetic study was determined by feeding (1-/sup 14/C)-sodium acetate to A. ustus cultures. The assignments made in the /sup 13/C-nmr spectrum of Austin are shown. The lowest dilution factor obtained in (1-/sup 14/C)-sodium acetate feeding experiments was 14. This dilution factor is sufficiently low to allow a successful feeding of (1,2-/sup 13/C/sub 2/)-sodium acetate. A new metabolite of A. ustus, deacetylaustin, was isolated and identified. An alkaloid of unknown structure was also isolated from the fungus.

  3. Sub-Inhibitory Concentrations of the Antibiotic Florfenicol Reduces Invasion in Isolates of Multi-Drug Resistant Salmonella Typhimurium DT104

    Science.gov (United States)

    Virulence can be enhanced in certain bacteria that are exposed to sub-lethal levels of antibiotics. Salmonella enterica serovar Typhimurium DT104 is resistant to five different antibiotics, including florfenicol. Using real-time PCR and a tissue culture invasion assay, we investigated the impact of ...

  4. The anticancer potential of steroidal saponin, dioscin, isolated from wild yam (Dioscorea villosa) root extract in invasive human breast cancer cell line MDA-MB-231 in vitro

    Science.gov (United States)

    Previously, we observed that wild yam (Dioscorea villosa) root extract (WYRE) was able to activate GATA3 in human breast cancer cells targeting epigenome. This study aimed to 'nd out if dioscin (DS), a bioactive compound of WYRE, can modulate GATA3 functions and cellular invasion in human breast can...

  5. Human platelets attenuate Aspergillus species via granule-dependent mechanisms.

    NARCIS (Netherlands)

    Perkhofer, S.; Kehrel, B.E.; Dierich, M.P.; Donnelly, J.P.; Nussbaumer, W.; Hofmann, J.; Eiff, C. von; Lass-Florl, C.

    2008-01-01

    Using laser scanning microscopy, we investigated whether platelets are capable of internalizing Aspergillus conidia and examined Aspergillus-platelet adherence. The influence of platelets on fungal growth was evaluated by assessing galactomannan (GM) release, hyphal elongation, and colony size. A

  6. Evolutionary Relationships among Aspergillus Species Producing Economically Important Mycotoxins

    Directory of Open Access Journals (Sweden)

    János Varga

    2003-01-01

    Full Text Available Several mycotoxins are produced by aspergilli, which contaminate our foods and feeds. Economically the most important of these mycotoxins are aflatoxins, ochratoxins and patulin. Recent studies indicate that these compounds can be produced by a number of unrelated Aspergillus species. Evolutionary relationships among sections involving these mycotoxin producing species (Aspergillus sections Circumdati, Flavi, Nigri, Fumigati and Clavati were evaluated using phylogenetic analysis of -tubulin and ribosomal RNA gene sequences. Phylogenetically unrelated species were found to produce the same mycotoxins. For example, aflatoxins have been produced under laboratory conditions by species belonging to three different sections, while ochratoxin A (OA and patulin have been produced by a variety of unrelated species. Based on this observation, mycotoxin-producing abilities of the isolates were lost (or gained several times during the evolution of the genus. Previous studies revealed that biosynthetic genes of several mycotoxins are clustered in the genome, leading to the idea that these clusters could be horizontally transferred as a unit to unrelated species, leading to the biosynthesis of the same mycotoxins in phylogenetically unrelated fungi. Such a mechanism could also explain the widespread occurrence of ochratoxin and patulin production in aspergilli. The organisation of biosynthetic genes of patulin and ochratoxins is unknown. Research is in progress in our laboratory to clarify the genetic background of biosynthesis of these mycotoxins, and to develop biosynthetic gene based probes for the molecular detection of these mycotoxin producing organisms.

  7. eNose technology can detect and classify human pathogenic molds in vitro: a proof-of-concept study of Aspergillus fumigatus and Rhizopus oryzae

    NARCIS (Netherlands)

    de Heer, K.; Vonk, S. I.; Kok, M.; Kolader, M.; Zwinderman, A. H.; van Oers, M. H. J.; Sterk, P. J.; Visser, C. E.

    2016-01-01

    Invasive pulmonary mold disease (IPMD) is often fatal in neutropenic patients. This is because IPMD is difficult to diagnose timely, especially when non-Aspergillus molds are the causative agent, as they are usually not associated with a positive galactomannan assay. In 2013 we showed that exhaled

  8. Comparative genomics of citric-acid-producing Aspergillus niger ATCC 1015 versus enzyme-producing CBS 513.88

    DEFF Research Database (Denmark)

    Andersen, Mikael Rørdam; Salazar, Margarita Pena; Schaap, Peter J.

    2011-01-01

    The filamentous fungus Aspergillus niger exhibits great diversity in its phenotype. It is found globally, both as marine and terrestrial strains, produces both organic acids and hydrolytic enzymes in high amounts, and some isolates exhibit pathogenicity. Although the genome of an industrial enzym...

  9. Azole resistance profile of amino acid changes in Aspergillus fumigatus CYP51A based on protein homology modeling.

    NARCIS (Netherlands)

    Snelders, E.; Karawajczyk, A.; Schaftenaar, G.; Verweij, P.E.; Melchers, W.J.G.

    2010-01-01

    Molecular studies have shown that the majority of azole resistance in Aspergillus fumigatus is associated with amino acid substitutions in the cyp51A gene. To obtain insight into azole resistance mutations, the cyp51A gene of 130 resistant and 76 susceptible A. fumigatus isolates was sequenced. Out

  10. Phenotypic differentiation of species from Aspergillus section Flavi on neutral red desiccated coconut agar

    DEFF Research Database (Denmark)

    Atanda, O. O.; Adetunji, M. C.; Ezekiel, C. N.

    2014-01-01

    In order to facilitate easy and rapid identification of aflatoxin-producing Aspergillus species, the phenotypic traits of Aspergillus section Flavi isolates were examined on neutral red desiccated coconut agar (NRDCA). Phenotype variations in colony morphology and the relationship between colour...... supported morphological differentiation of the four species based on colony features, conidia type and colour. In particular, the two very closely related minisclerotial species, A. minisclerotigenes and A. parvisclerotigenus, were clearly differentiated by their colony colour on NRDCA. All toxigenic...... isolates produced aflatoxins in the culture medium in varying quantities. Plates of aflatoxigenic A. flavus L strains fluoresced bluish purple/lavender around the colony on the obverse and pastel blue on the reverse side due to aflatoxin B production while those of A. minisclerotigenes, A. parasiticus...

  11. SECONDARY METABOLITE FROM ENDOPHYTIC FUNGI Aspergillus niger OF THE STEM BARK OF KANDIS GAJAH (Garcinia griffithii

    Directory of Open Access Journals (Sweden)

    Elfita Elfita

    2012-06-01

    Full Text Available Garcinia griffithii are known as kandis gajah including the Garcinia genus. This plant has been traditionally used by local communities Sarasah Bonta, Lembah Arau, West Sumatra, to treat various diseases including gout. Aspergillus niger was isolated from the tissues of the stem bark of Garcinia griffithii. The fungi strain was identified base on colony and cell morphology characteristic. Aspergillus niger cultured in media 5L Potatos Dextose Broth (PDB for 8 weeks and filtered. Media that already contains secondary metabolites are partitioned using ethyl acetate solvent in 5 L (twice, followed by evaporation. Furthermore, the extract is separated by chromatographic techniques to obtain a pure compound of white crystal. The molecular structures of isolated compounds are determined by spectroscopic methods including IR, 1H-NMR, 13C-NMR, HMQC, HMBC, and COSY. The compound was determined as phenolic (1.

  12. Motuporamines, anti-invasion and anti-angiogenic alkaloids from the marine sponge Xestospongia exigua (Kirkpatrick): isolation, structure elucidation, analogue synthesis, and conformational analysis.

    Science.gov (United States)

    Williams, David E; Craig, Kyle S; Patrick, Brian; McHardy, Lianne M; van Soest, Rob; Roberge, Michel; Andersen, Raymond J

    2002-01-11

    Extracts of the sponge Xestospongia exigua collected in Papua New Guinea were positive in a new assay for anti-invasion activity. Bioassay-guided fractionation led to the identification of the three known motuporamines A (1), B (2), and C (3) along with the new motuporamines D (4), E (5), and F (6) and a mixture of G, H, and I (15). Motuporamines A (1), B (2), and C (3) and the mixture of G, H, and I (15) were responsible for the anti-invasion activity of the crude extract. Motuporamine C (3) has also been found to be anti-angiogenic. A series of analogues of the motuporamines have been synthesized and evaluated for anti-invasive activity. These SAR results revealed that a saturated 15-membered cyclic amine fused to the natural motuporamine diamine side chain (13) represented the optimal structure for anti-invasive activity in this family. Single-crystal X-ray diffraction analysis of one of the analogues 20 showed that in the solid state its 16-membered macrocyclic amine fragment adopted the [4444] quadrangular conformation predicted by calculations to be the lowest energy conformation for the corresponding cycloalkane, cyclohexadecane. These data along with literature X-ray data and conformational analysis for derivatives of azacyclotridecane have been used as precedents for predicting the lowest energy ring conformations of other motuporamines. The SAR data from the natural and synthetic motuporamines have been combined with the conformational analyses to provide an outline of the functionality and shape required for activity in this family of alkaloids and to design a new analogue 49 that showed good anti-invasion activity.

  13. Aspergillus infection of the respiratory tract after lung transplantation: chest radiographic and CT findings

    International Nuclear Information System (INIS)

    Diederich, S.; Scadeng, M.; Flower, C.D.R.; Dennis, C.; Stewart, S.

    1998-01-01

    The objective of our study was to assess radiographic and CT findings in lung transplant patients with evidence of Aspergillus colonization or infection of the airways and correlate the findings with clinical, laboratory, bronchoalveolar lavage, biopsy and autopsy findings. The records of 189 patients who had undergone lung transplantation were retrospectively reviewed for evidence of Aspergillus colonization or infection of the airways. Aspergillus was demonstrated by culture or microscopy of sputum or bronchoalveolar lavage fluid or histologically from lung biopsies or postmortem studies in 44 patients (23 %). Notes and radiographs were available for analysis in 30 patients. In 12 of the 30 patients (40 %) chest radiographs remained normal. In 11 of 18 patients with abnormal radiographs pulmonary abnormalities were attributed to invasive pulmonary aspergillosis (IPA) in the absence of other causes for pulmonary abnormalities (8 patients) or because of histological demonstration of IPA (3 patients). In these 11 patients initial radiographic abnormalities were focal areas of patchy consolidation (8 patients), ill-defined pulmonary nodules (2 patients) or a combination of both (1 patient). In some of the lesions cavitation was demonstrated subsequently. At CT a ''halo'' of decreased density was demonstrated in some of the nodules and lesion morphology and location were shown more precisely. Demonstration of Aspergillus from the respiratory tract after lung transplantation does not necessarily reflect IPA but may represent colonization of the airways or semi-invasive aspergillosis. The findings in patients with IPA did not differ from those described in the literature in other immunocompromised patients, suggesting that surgical disruption of lymphatic drainage and nervous supply or effects of preservation and transport of the transplant lung do not affect the radiographic appearances. (orig.)

  14. In vitro interaction of voriconazole and anidulafungin against triazole-resistant Aspergillus fumigatus.

    Science.gov (United States)

    Seyedmousavi, Seyedmojtaba; Meletiadis, Joseph; Melchers, Willem J G; Rijs, Antonius J M M; Mouton, Johan W; Verweij, Paul E

    2013-02-01

    Voriconazole is the recommended drug of first choice to treat infections caused by Aspergillus fumigatus. The efficacy of voriconazole might be hampered by the emergence of azole resistance. However, the combination of voriconazole with anidulafungin could improve therapeutic outcomes in azole-resistant invasive aspergillosis (IA). The in vitro interaction between voriconazole and anidulafungin was determined against voriconazole-susceptible and voriconazole-resistant (substitutions in the cyp51A gene, including single point [M220I and G54W] and tandem repeat [34-bp tandem repeat in the promoter region of the cyp51A gene in combination with substitutions at codon L98 and 46-bp tandem repeat in the promoter region of the cyp51A gene in combination with mutation at codons Y121 and T289] mutations) clinical A. fumigatus isolates using a checkerboard microdilution method with spectrophotometric analysis and a viability-based XTT {2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide} assay within 2 h of exposure after 24 and 48 h of incubation at 35 °C to 37 °C. Fractional inhibitory concentration (FIC) indexes (FICis) were determined using different MIC endpoints and Bliss independence analysis performed based on the response surface calculation of the no-drug interaction. Significant synergistic interactions obtained based on measuring the FIC index were dependent on the MIC endpoint, in which FICs were inversely related to voriconazole and anidulafungin MICs and were influenced by the CYP51A genotype. A statistically significant difference was observed between FIC indexes of isolates harboring tandem repeat mutations and wild-type controls (P = 0.006 by one-way analysis of variance [ANOVA]), indicating that synergy is decreased in azole-resistant strains. Our results indicated that a combination of voriconazole and anidulafungin might be effective against infections caused by both azole-susceptible and azole-resistant A

  15. Successful treatment of azole-resistant invasive aspergillosis in a bottlenose dolphin with high-dose posaconazole

    NARCIS (Netherlands)

    P.E. Bunskoek (Paulien); S. Seyedmousavi (Seyedmojtaba); S. Gans (Steven); van Vierzen, P.B.J. (Peter B.J.); W.J. Melchers (Willem); C.E. van Elk; J.W. Mouton (Johan); P.E. Verweij (Paul)

    2017-01-01

    textabstractInvasive aspergillosis due to azole-resistant Aspergillus fumigatus is difficult to manage. We describe a case of azole-resistant invasive aspergillosis in a female bottlenose dolphin, who failed to respond to voriconazole and posaconazole therapy. As intravenous therapy was precluded,

  16. Aspiperidine oxide, a piperidine N-oxide from the filamentous fungus Aspergillus indologenus

    DEFF Research Database (Denmark)

    Petersen, Lene Maj; Kildgaard, Sara; Jaspars, Marcel

    2015-01-01

    A novel secondary metabolite, aspiperidine oxide, was isolated from the filamentous fungus, Aspergillus indologenus. The structure of aspiperidine oxide was determined from extensive 1D and 2D NMR spectroscopic analysis supported by high-resolution mass spectrometry. The structure revealed a rare...... piperidine N-oxide, not observed in filamentous fungi before. A biosynthetic pathway towards aspiperidine oxide is proposed, based on tentative identification of intermediates from UHPLC-DAD-HRMS data....

  17. Identification of toxigenic Aspergillus species from diet dairy goat using a polyphasic approach

    OpenAIRE

    Silva, Janaina Lara da; Aparecido, Christiane Ceriani; Hansen, Daiane; Pereira, Tacila Alves Muniz; Felicio, Joana D'arc; Gonçalez, Edlayne

    2015-01-01

    Some species of filamentous fungi that infest agricultural commodities are able to produce mycotoxins, contaminating feed and animal products. The aim of this research was to identify the mycoflora present in the feed and forage for dairy goat and to isolate and characterize the Aspergillus flavus and A. parasiticus strains based on a morphological and molecular characterization and mycotoxigenic ability. The goat dairy diets were collected monthly from 11 goat milk farms, totaling 129 and 10...

  18. Development of a green fluorescent tagged strain of Aspergillus carbonarius to monitor fungal colonization in grapes

    OpenAIRE

    Crespo Sempere, Ana; López Pérez, Mario; Martínez-Culebras, Pedro V.; González-Candelas, Luis

    2011-01-01

    An enhanced green fluorescent protein has been used to tag an OTA-producing strain of Aspergillus carbonarius (W04-40) isolated from naturally infected grape berries. Transformation of the fungus was mediated by Agrobacterium tumefaciens. The most efficient transformation occurred when the co-cultivation was done with 104 conidia due to higher frequency of resistance colonies (894 per 104 conidia) and lower background obtained. To confirm the presence of the hph gene in hygromycin resistant c...

  19. Methanol utilization by a strain of Aspergillus niger: influence on the synthesis and activity of pectinases.

    Science.gov (United States)

    Maldonado, M C; Strasser de Saad, A M; Callieri, D A

    1993-03-01

    During the production of pectinases by a strain of Aspergillus niger isolated from rotten lemons, methanol was liberated into the medium due to the cleavage of the pectin molecule used as the carbon source. The methanol was subsequently consumed by the microorganism but neither the synthesis nor the activity of pectinesterase and polygalacturonase was affected. Although not studied in detail, the mechanism involved in the utilization of methanol is similar to that described for methylotrophic yeasts.

  20. Neutrophil chemotactic responses induced by fresh and swollen Rhizopus oryzae spores and Aspergillus fumigatus conidia.

    OpenAIRE

    Waldorf, A R; Diamond, R D

    1985-01-01

    With the induction of germination, Rhizopus oryzae spores and Aspergillus fumigatus conidia activate the complement system and induce neutrophil chemotaxis. In contrast, freshly isolated R. oryzae spores did not induce neutrophil migration into lung tissue of mice after intranasal inoculation. Moreover, in microchemotaxis assays neither fresh R. oryzae spores nor A. fumigatus conidia activated sera to stimulate human neutrophil chemotaxis above control migration until at least 10(7) or 10(8) ...