DOCSLIB.ORG

Genetic Variability, Antigenicity and Antifungal Susceptibility of Madurella Mycetomatis

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Genetic Variability, Antigenicity and Antifungal Susceptibility of Madurella Mycetomatis Genetic Variability, Antigenicity and Antifungal Susceptibility of Madurella mycetomatis ISBN 978-90-8559-311-9 © Wendy van de Sande, 2007 All rights reserved. No part of this thesis may be reproduced or transmitted in any form, by any means, electronic or mechanical, without the written permission of the author, or where appropriate the publisher of the articles Printed by: Optima Grafische Communicatie, Rotterdam, The Netherlands Graphic design: Wendy van de Sande Genetic Variability, Antigenicity and Antifungal Susceptibility of Madurella mycetomatis Genetische variabiliteit, antigeniciteit en antifungale gevoeligheid van Madurella mycetomatis Proefschrift ter verkrijging van de graad van doctor aan de Erasmus Universiteit Rotterdam op gezag van de rector magnificus Prof.dr. S.W.J. Lamberts en volgens besluit van het College voor Promoties De openbare verdediging zal plaatsvinden op: Woensdag 24 oktober 2007 om 13:45 uur Door Wendy Wilhelmina Johanna van de Sande geboren te Breda Promotiecommissie Promotor: Prof.dr.dr. A.F. van Belkum Overige leden: Prof.dr. G.S. de Hoog Prof.dr. A.P. Oranje Prof.dr. H.A. Verbrugh Co-promotor: Dr. I.A.J.M. Bakker-Woudenberg The printing of this thesis was financially supported by: BD Diagnostic Systems, bioMérieux Benelux B.V., Eurogentec B.V., Janssen-Cilag, Merck Sharp & Dohme, Novartis Pharma and Pfizer B.V. Contents General introduction Chapter 1 Scope of the thesis 11 Chapter 2 Black grain myetoma, a neglected infection 15 Genetic variation in Madurella mycetomatis and its human host Chapter 3 Genotyping of Madurella mycetomatis by selective 39 amplification of restriction fragments (AFLP®) and subtype-correlation with geographical origin and lesion size Chapter 4 Polymorphisms in genes involved in innate immunity 53 predispose towards mycetoma susceptibility Antigenicity of Madurella mycetomatis Chapter 5 Translationally controlled tumour protein from 73 Madurella mycetomatis, a marker for tumorous mycetoma progression Chapter 6 Madurella mycetomatis compounds cross-reactive 93 with galactomannan are detectable in culture supernatant but not in serum Antifungal susceptibility of Madurella mycetomatis Chapter 7 In vitro susceptibilities of Madurella mycetomatis 101 to itraconazole and amphotericin B assessed by a modified NCCLS method and a viability-based 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5- [(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) assay Chapter 8 In vitro antifungal susceptibility testing of 109 Madurella mycetomatis for six antifungal agents by the Sensititre system in comparison with a viability-based XTT assay and a modified NCCLS method Chapter 9 In vitro susceptibility of Madurella mycetomatis, 119 prime agent of madura foot, to tea tree oil and artemisinin Chapter 10 Melanin biosynthesis in Madurella mycetomatis 127 and its effect on susceptibility to itraconazole and ketoconazole General discussion and concluding remarks Chapter 11 Summarizing discussion 145 Chapter 12 Nederlandse samenvatting 159 Chapter 13 Dankwoord 175 Chapter 14 List of publications and curriculum vitae 179 General introduction Chapter 1 Scope of the thesis Wendy W.J. van de Sande Chapter 1 12 Scope of the thesis Introduction Mycetoma is a chronic, granulomatous, progressive inflammatory disease, characterized by a subcutaneous mass, tumefaction and the formation of sinus tracts. The sinuses discharge seropurulent material and fungal or bacterial grains. This disease, which can be caused by both bacteria and fungi, is encountered all over the world but is endemic in the so-called “mycetoma-belt”, between 30 oN and 15 oS of the equator. Sudan is one of the countries in which mycetoma has a high incidence. In this country the most common causative agent is the fungus Madurella mycetomatis. Although mycetoma was already described in 1842, still relatively little is known about this disease. Diagnostic assays and treatment are still suboptimal. Current diagnostic assays are time-consuming and not very reliable. Treatment, especially in eumycetoma, is difficult and usually involves both surgery and medical treatment. Adequate antifungal treatment may take years and is usually started without even knowing if the causative agent is actually susceptible to the chemotherapeutic agent applied. Scope of the thesis To understand the difficulties in diagnosing and treating mycetoma we first review this disease in chapter 2. The current knowledge about this disease, its prevelance, the diagnostic assays used, and the treatment given to the patients are highlighted. As will become clear in this chapter little is known about the disease. In the remaining of this thesis we will focus on three main themes: 1. Genetic variation of M. mycetomatis and its human host In the chapters belonging to this theme it was assessed whether genetic variability of both pathogen and host could be related to the outcome of the disease. In chapter 3 the selective amplification of restriction fragments (AFLP®) is used to devide 37 Sudanese clinical isolates of M. mycetomatis into two main clusters (clusters I and II) and one minor cluster (cluster III). The strains in cluster I mostly derived from patients in Central Sudan and were generally obtained from larger lesions than the strains found in cluster II. In chapter 4 several human genes involved in neutrophil function were studied with PCR-restriction analysis. It appeared that patients usually had a genotype correlated with a higher CXCL8 expression and a lower nitric oxide expression. This was confirmed phenotypically. 2. Antigenicity of M. mycetomatis In order to improve the diagnostic serology for mycetoma caused by M. mycetomatis the antigenicity of M. mycetomatis was assessed. In chapter 5 the first immunogenic antigen, a protein homologous to the translationally controlled tumour protein (TCTP) was discovered using a M. mycetomatis cDNA expression library. Both IgM and IgG responses could be measured for this protein, with the highest antibody levels found associated with the largest 13 Chapter 1 lesions. Unfortunately, not all patients formed antibodies against this protein. In chapter 6 it was shown that although M. mycetomatis secreted significant amounts of galactomannan-like compounds into the culture medium, these compounds were not detected in the patient serum. 3. Antifungal susceptibility of M. mycetomatis In order to improve the antifungal therapy in patients we tried to gain more insight in the in vitro susceptibility of M. mycetomatis against various antifungal agents. In chapter 7 a CLSI (formerly NCCLS) based assay was developed for M. mycetomatis. To facilitate endpoint reading a 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H- tetrazolium hydroxide (XTT) assay for fungal viability was used. In chapter 8 the assay described in chapter 7 was compared to a commercial Sensititre assay for the antifungal agents amphotericin B, ketoconazole, itraconazole, fluconazole and voriconazole. In chapter 9 the in vitro susceptibilities for the less commonly used antifungal agents artemisinin and tea tree oil were determined. In chapter 10 it was assessed what the effect of melanin produced by M. mycetomatis was on the in vitro susceptibilities to amphotericin B, ketoconazole, itraconazole, fluconazole and voriconazole. Finally in chapters 11 and 12 the findings reported in this thesis are summarized and discussed. 14 Chapter 2 Black grain mycetoma, a neglected infection Wendy W.J. van de Sande Adapted from: Abdalla O.A. Ahmed, Willem van Leeuwen, Ahmed Fahal, Wendy W.J. van de Sande, Henri Verbrugh and Alex van Belkum, Mycetoma caused by Madurella mycetomatis: a neglected infectious burden, The Lancet Infectious Diseases 2004, 4: 566-574. Abdalla O.A. Ahmed, Wendy W.J. van de Sande, Ahmed Fahal, Irma Bakker-Woudenberg, Henri Verbrugh and Alex van Belkum, Management of Mycetoma: Major Challenge in Tropical Mycoses with Limited International Recognition, Current Opinion in Infectious Diseases 2007, 20(7): 146-151. Chapter 2 16 Black grain mycetoma, a neglected infection Introduction Mycetoma is a chronic infectious disease, which remains localized, involves cutaneous and subcutaneous tissues, the fascia and bones (95). The disease is characterized by tumefaction, draining sinuses and the presence of grains (95). The oldest recorded case of possible mycetoma involves the skeleton found in Israel of an adult woman buried in the Byzantine period (300-600 AD) (95). This skeleton had morphologic changes in the bones of the distal portion of the lower extremities which are suggestive for mycetoma (95). The first written description of mycetoma was found in the Indian religious book Atharva Veda, where the disease was called “padavalmika”, meaning foot-anthill (95). In 1842 the first case was described in western literature by Dr. John Gill, a physician of the Madras Medical Service of the British Army in India (112). His colleague Dr. Colebrook proposed the name “Madura foot” in 1848 (95, 112). The term mycetoma was first used by Vandyke Carter in 1860 (112). He was the first to believe that mycetoma could be separated into two varieties depending on the colour of the grains associated with the disease. M. mycetomatis probably caused the black variety of mycetoma Carter described, whereas one of the aerobic actinomycetes caused the light-coloured variety (95). Epidemiology and causation As was already apparent from this short history lesson, mycetoma has a worldwide but uneven distribution. The disease is endemic in tropical and subtropical
Load more

Recommended publications
  • Revision of Agents of Black-Grain Eumycetoma in the Order Pleosporales
    Persoonia 33, 2014: 141–154 www.ingentaconnect.com/content/nhn/pimj RESEARCH ARTICLE http://dx.doi.org/10.3767/003158514X684744 Revision of agents of black-grain eumycetoma in the order Pleosporales S.A. Ahmed1,2,3, W.W.J. van de Sande 4, D.A. Stevens 5, A. Fahal 6, A.D. van Diepeningen 2, S.B.J. Menken 3, G.S. de Hoog 2,3,7 Key words Abstract Eumycetoma is a chronic fungal infection characterised by large subcutaneous masses and the pres- ence of sinuses discharging coloured grains. The causative agents of black-grain eumycetoma mostly belong to the Madurella orders Sordariales and Pleosporales. The aim of the present study was to clarify the phylogeny and taxonomy of mycetoma pleosporalean agents, viz. Madurella grisea, Medicopsis romeroi (syn.: Pyrenochaeta romeroi), Nigrograna mackin­ Pleosporales nonii (syn. Pyrenochaeta mackinnonii), Leptosphaeria senegalensis, L. tompkinsii, and Pseudochaetosphaeronema taxonomy larense. A phylogenetic analysis based on five loci was performed: the Internal Transcribed Spacer (ITS), large Trematosphaeriaceae (LSU) and small (SSU) subunit ribosomal RNA, the second largest RNA polymerase subunit (RPB2), and transla- tion elongation factor 1-alpha (TEF1) gene. In addition, the morphological and physiological characteristics were determined. Three species were well-resolved at the family and genus level. Madurella grisea, L. senegalensis, and L. tompkinsii were found to belong to the family Trematospheriaceae and are reclassified as Trematosphaeria grisea comb. nov., Falciformispora senegalensis comb. nov., and F. tompkinsii comb. nov. Medicopsis romeroi and Pseu­ dochaetosphaeronema larense were phylogenetically distant and both names are accepted. The genus Nigrograna is reduced to synonymy of Biatriospora and therefore N.
    [Show full text]
  • A Comparative Study of in Vitro Susceptibility of Madurella
    Original Article A comparative Study of In vitro Susceptibility of Madurella mycetomatis to Anogeissus leiocarpous Leaves, Roots and Stem Barks Extracts Ikram Mohamed Eltayeb*1, Abdel Khalig Muddathir2, Hiba Abdel Rahman Ali3 and Saad Mohamed Hussein Ayoub1 1Department of Pharmacognosy, Faculty of Pharmacy, University of Medical Sciences and Technology, P. O. Box 12810, Khartoum, Sudan 2Department of Pharmacognosy, Faculty of Pharmacy, University of Khartoum, Khartoum, Sudan 3Commission of Biotechnology and Genetic Engineering, National Center for Research, Khartoum, Sudan ABSTRACT Objective: Anogeissus leiocarpus leaves, roots and stem bark are broadly utilized as a part of African traditional medicine against numerous pathogenic microorganisms for treating skin diseases and infections. Mycetoma disease is a fungal and/ or bacterial skin infection, mainly caused by filamentous Madurella mycetomatis fungus. The objective of this study is to investigate and compare the antifungal activity of A. leiocarpus leaves, roots and stem bark against the isolated mycetoma pathogen, M. mycetomatis fungus. Methods: The alcoholic crude extracts, and their petroleum ether, chloroform and ethyl acetate fractions of A. leiocarpus leaves, roots and stem bark were prepared and their antifungal activity against the isolated M. mycetomatis fungus were assayed according to the Address for NCCLS antifungal modified method and MTT assay compared to the Correspondence Ketoconazole, standard antifungal drug. The most bioactive fractions were subjected to chemical analysis using LC-MS/MS Department of chromatographic analytical method. Pharmacognosy, Results: The results demonstrated the potent antifungal activity of A. Faculty of Pharmacy, leiocarpus extracts against the isolated pathogenic M. mycetomatis University of Medical compared to the negative and positive controls. The chloroform Sciences and fractions showed higher antifungal activity among the other extracts, Technology, P.
    [Show full text]
  • Utilization of the Internal Transcribed Spacer Regions As Molecular Targets
    Medical Mycology 2002, 40, 87±109 Accepted 9July 2001 Review article Utilizationof the internaltranscribed spacer regions as molecular targets to detect andidentify human fungal pathogens P.C.IWEN*, S.H.HINRICHS* & M.E.RUPP Downloaded from https://academic.oup.com/mmy/article/40/1/87/961355 by guest on 29 September 2021 y *Department ofPathology and Microbiology,University ofNebraska MedicalCenter, Omaha, Nebraska, USA; Internal Medicine, y University ofNebraska MedicalCenter, Omaha, Nebraska, USA Advancesin molecular technology show greatpotential for the rapiddetection and identication of fungifor medical,scienti c andcommercial purposes. Numerous targetswithin the fungalgenome have been evaluated, with much of the current work usingsequence areas within the ribosomalDNA (rDNA) gene complex. This sectionof the genomeincludes the 18S,5 8Sand28S genes which codefor ribosomal ¢ RNA(rRNA) andwhich havea relativelyconserved nucleotide sequence among fungi.It alsoincludes the variableDNA sequence areas of the interveninginternal transcribedspacer (ITS) regionscalled ITS1 and ITS2. Although not translatedinto proteins,the ITScoding regions have a criticalrole in the developmentof functional rRNA,with sequencevariations among species showing promiseas signature regionsfor molecularassays. This review of the current literaturewas conducted to evaluateclinical approaches for usingthe fungalITS regions as molecular targets. Multipleapplications using the fungalITS sequences are summarized here including those for cultureidenti cation, phylogenetic
    [Show full text]
  • New Species of Madurella, Causative Agents of Black-Grain Mycetoma
    New Species of Madurella, Causative Agents of Black-Grain Mycetoma G. Sybren de Hoog,a,b,c,d Anne D. van Diepeningen,a El-Sheikh Mahgoub,e and Wendy W. J. van de Sandef Centraalbureau voor Schimmelcultures KNAW Fungal Biodiversity Centre, Utrecht, The Netherlandsa; Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam, Amsterdam, The Netherlandsb; Peking University Health Science Center, Research Center for Medical Mycology, Beijing, Chinac; Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou, Chinad; Mycetoma Research Centre, University of Khartoum, Khartoum, Sudane; and Erasmus Medical Center, Department of Medical Microbiology and Infectious Diseases, Rotterdam, The Netherlandsf Downloaded from A new species of nonsporulating fungus, isolated in a case of black-grain mycetoma in Sudan, is described as Madurella fahalii. The species is characterized by phenotypic and molecular criteria. Multigene phylogenies based on the ribosomal DNA (rDNA) internal transcribed spacer (ITS), the partial ␤-tubulin gene (BT2), and the RNA polymerase II subunit 2 gene (RPB2) indicate that M. fahalii is closely related to Madurella mycetomatis and M. pseudomycetomatis; the latter name is validated according to the rules of botanical nomenclature. Madurella ikedae was found to be synonymous with M. mycetomatis. An isolate from Indo- nesia was found to be different from all known species based on multilocus analysis and is described as Madurella tropicana. Madurella is nested within the order Sordariales, with Chaetomium as its nearest neighbor. Madurella fahalii has a relatively low optimum growth temperature (30°C) and is less susceptible to the azoles than other Madurella species, with voriconazole http://jcm.asm.org/ and posaconazole MICs of 1 ␮g/ml, a ketoconazole MIC of 2 ␮g/ml, and an itraconazole MIC of >16 ␮g/ml.
    [Show full text]
  • The Discovery of Fenarimols As Novel Drug
    bioRxiv preprint doi: https://doi.org/10.1101/258905; this version posted February 2, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. Full title: Addressing the Most Neglected Diseases through an Open Research Model: the Discovery of Fenarimols as Novel Drug Candidates for Eumycetoma. Short title: Eumycetoma Open Research Model Drug discovery 1 bioRxiv preprint doi: https://doi.org/10.1101/258905; this version posted February 2, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Addressing the Most Neglected Diseases through an Open Research Model: the Discovery of 2 Fenarimols as Novel Drug Candidates for Eumycetoma. 3 4 Wilson Lim1, Youri Melse1, Mickey Konings1, Hung Phat Duong2, Kimberly Eadie1, Benoît Laleu3, Ben 4 2 4 1 5 Perry , Matthew H. Todd , Jean-Robert Ioset , Wendy W.J. van de Sande * 6 7 8 1Erasmus MC, Department of Medical Microbiology and Infectious Diseases, Rotterdam, The 9 Netherlands 10 2School of Chemistry, The University of Sydney, Sydney, Australia 11 3Medicines for Malaria Venture (MMV), Geneva, Switzerland 12 4DNDi, Geneva, Switzerland 13 14 15 16 Corresponding author: Wendy W.J. van de Sande 17 * [email protected] (WvdS) 2 bioRxiv preprint doi: https://doi.org/10.1101/258905; this version posted February 2, 2018.
    [Show full text]
  • Niclosamide Is Active in Vitro Against Mycetoma Pathogens
    molecules Article Niclosamide Is Active In Vitro against Mycetoma Pathogens Abdelhalim B. Mahmoud 1,2,3,† , Shereen Abd Algaffar 4,† , Wendy van de Sande 5, Sami Khalid 4 , Marcel Kaiser 1,2 and Pascal Mäser 1,2,* 1 Department of Medical Parasitology and Infection Biology, Swiss Tropical and Public Health Institute, 4051 Basel, Switzerland; [email protected] (A.B.M.); [email protected] (M.K.) 2 Faculty of Science, University of Basel, 4001 Basel, Switzerland 3 Faculty of Pharmacy, University of Khartoum, Khartoum 11111, Sudan 4 Faculty of Pharmacy, University of Science and Technology, Omdurman 14411, Sudan; [email protected] (S.A.A.); [email protected] (S.K.) 5 Erasmus Medical Center, Department of Medical Microbiology and Infectious Diseases, 3000 Rotterdam, The Netherlands; [email protected] * Correspondence: [email protected]; Tel.: +41-61-284-8338 † These authors contributed equally to this work. Abstract: Redox-active drugs are the mainstay of parasite chemotherapy. To assess their repurposing potential for eumycetoma, we have tested a set of nitroheterocycles and peroxides in vitro against two isolates of Madurella mycetomatis, the main causative agent of eumycetoma in Sudan. All the tested compounds were inactive except for niclosamide, which had minimal inhibitory concentrations of around 1 µg/mL. Further tests with niclosamide and niclosamide ethanolamine demonstrated in vitro activity not only against M. mycetomatis but also against Actinomadura spp., causative agents of actinomycetoma, with minimal inhibitory concentrations below 1 µg/mL. The experimental compound MMV665807, a related salicylanilide without a nitro group, was as active as niclosamide, indicating that the antimycetomal action of niclosamide is independent of its redox chemistry (which Citation: Mahmoud, A.B.; Abd Algaffar, S.; van de Sande, W.; Khalid, is in agreement with the complete lack of activity in all other nitroheterocyclic drugs tested).
    [Show full text]
  • Sunday 1 April Parallel Session 5: Mitochondria
    PS4.8 How do mobile pathogenicity chromosomes collaborate with the core genome? Charlotte van der Does, Martijn Rep University of Amsterdam In the tomato pathogen F. oxysporum f. sp. lycopersici, most known effector genes reside on a pathogenicity chromosome that can be exchanged between strains through horizontal transfer. As a result, this fungus has sub‐ genomes with different evolutionary histories: the conserved core genome and the mobile ‘extra’ genome. Interestingly, expression of the effectors on the mobile genome requires Sge1, a conserved transcription factor encoded in the core genome. Also, a transcription factor on the mobile chromosome, Ftf1, is associated with pathogenicity (de Vega‐Bartol et al. 2011). Random insertion of an effector‐promoter GFP reporter construct revealed that the position in the genome has a strong influence on the level of expression. Furthermore, in all transformants tested, expression could no longer be induced, but was constitutive. To discover how Sge1 and Ftf1 control gene expression from the core genome and the mobile chromosome(s), their targets will be identified using ChIPseq: sequencing of DNA fragments to which Sge1 binds and RNAseq: in depth‐sequencing of transcripts, comparing standard culture conditions to in planta‐mimicing conditions. Additional components necessary to express effector genes on mobile chromosomes will be identified via insertional mutagenesis. Sunday 1 April Parallel session 5: Mitochondria PS5.1 Functional analysis of ERMES and TOB (SAM) complex components in Neurospora crassa. Frank E. Nargang, Sebastian W.K. Lackey, Jeremy G. Wideman Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9 The Neurospora crassa TOB complex (topogenesis of beta‐barrel proteins), also known as the SAM complex (sorting and assembly machinery), inserts a subset of mitochondrial outer membrane proteins into the membrane—including all beta‐barrel proteins.
    [Show full text]
  • The Development of a Novel Diagnostic PCR for Madurella Mycetomatis Using a Comparative Genome Approach
    PLOS NEGLECTED TROPICAL DISEASES RESEARCH ARTICLE The development of a novel diagnostic PCR for Madurella mycetomatis using a comparative genome approach 1 1,2 1 1 Wilson LimID , Emmanuel SiddigID , Kimberly Eadie , Bertrand NyuykongeID , 3¤a¤b 2 1 4 Sarah Ahmed , Ahmed Fahal , Annelies Verbon , Sandra SmitID , Wendy WJ van de 1 SandeID * 1 Erasmus MC, University Medical Center Rotterdam, Department of Microbiology and Infectious Diseases, a1111111111 Rotterdam, The Netherlands, 2 Mycetoma Research Centre, University of Khartoum, Khartoum, Sudan, a1111111111 3 Westerdijk Fungal Biodiversity Institute, Utrecht, The Netherlands, 4 Wageningen University & Research, a1111111111 Department of Plant Science, Wageningen, The Netherlands a1111111111 ¤a Current address: Center of Expertise in Mycology of Radboud University Medical Center / Canisius a1111111111 Wilhelmina Hospital, Nijmegen, The Netherlands ¤b Current address: Faculty of medical laboratory sciences, University of Khartoum, Sudan * [email protected] OPEN ACCESS Abstract Citation: Lim W, Siddig E, Eadie K, Nyuykonge B, Ahmed S, Fahal A, et al. (2020) The development of a novel diagnostic PCR for Madurella mycetomatis using a comparative genome approach. PLoS Negl Background Trop Dis 14(12): e0008897. https://doi.org/ Eumycetoma is a neglected tropical disease most commonly caused by the fungus Madur- 10.1371/journal.pntd.0008897 ella mycetomatis. Identification of eumycetoma causative agents can only be reliably per- Editor: Husain Poonawala, Lowell General Hospital, formed by molecular identification, most commonly by species-specific PCR. The current M. UNITED STATES mycetomatis specific PCR primers were recently discovered to cross-react with Madurella Received: June 18, 2020 pseudomycetomatis. Here, we used a comparative genome approach to develop a new M.
    [Show full text]
  • Drivers of Genetic Diversity in Secondary Metabolic Gene Clusters in a Fungal Population 5 6 7 8 Abigail L
    bioRxiv preprint doi: https://doi.org/10.1101/149856; this version posted July 11, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 2 3 4 Drivers of genetic diversity in secondary metabolic gene clusters in a fungal population 5 6 7 8 Abigail L. Lind1, Jennifer H. Wisecaver2, Catarina Lameiras3, Philipp Wiemann4, Jonathan M. 9 Palmer5, Nancy P. Keller4, Fernando Rodrigues6,7, Gustavo H. Goldman8, Antonis Rokas1,2 10 11 12 1. Department of Biomedical Informatics, Vanderbilt University School of Medicine, Nashville, 13 Tennessee, USA. 14 2. Department of Biology, Vanderbilt University, Nashville, Tennessee, USA. 15 3. Department of Microbiology, Portuguese Oncology Institute of Porto, Porto, Portugal 16 4. Department of Medical Microbiology & Immunology, University of Wisconsin-Madison, 17 Madison, Wisconsin, USA 18 5. Center for Forest Mycology Research, Northern Research Station, US Forest Service, Madison, 19 Wisconsin, USA 20 6. Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, 21 Braga, Portugal 22 7. ICVS/3B's - PT Government Associate Laboratory, Braga/Guimarães, Portugal. 23 8. Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, São 24 Paulo, Brazil 25 †Corresponding author and lead contact: [email protected] 26 bioRxiv preprint doi: https://doi.org/10.1101/149856; this version posted July 11, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.
    [Show full text]
  • Madurella Grisea: a Case Report on an Uncommon Cause for Mycetoma
    127 Case Report Madurella grisea: a case report on an uncommon cause for mycetoma MN Jayawardena1, NNTM Wickremasinghe2, PI Jayasekera1 Sri Lankan Journal of Infectious Diseases 2017 Vol. 7(2):127-130 DOI: http://dx.doi.org/10.4038/sljid.v7i2.8140 Abstract Madura foot is a disfiguring, disabling condition, found in the tropics and subtropics. We present a case of a farmer who presented with swelling and multiple discharging sinuses in his left foot. The offending organism, Madurella grisea was identified by fungal culture. The patient was managed with local excision combined with systemic antifungal therapy. So far, the patient has made an uneventful recovery, with no recurrences. The laboratory diagnosis proved difficult, given the limited resources. We wish to highlight the difficulties faced in diagnosis and management of this condition, as response to antifungals are dependent on the pathogen, and adherence to long term antifungals are crucial. Keywords: mycetoma, Madurella grisea, madura foot Introduction Mycetoma is a destructive, chronic inflammatory disease, mainly affecting the feet, with the population at risk being those who are socioeconomically disadvantaged, such as labourers and agriculturalists. Sri Lankan data regarding the aetiological agents of this disease is sparse. The control of this neglected tropical disease requires better knowledge and understanding by the clinicians, continuous supply of good quality antifungal agents and high quality fungal diagnostic facilities. Case report A 55 year old previously well farmer presented to Base Hospital, Mahiyanganaya, complaining of painless swelling of his left ankle of 6 months duration, which was insidious in onset. There were no signs of systemic disease.
    [Show full text]
  • Enhanced Fungal Isolation by Use of a Novel Technique
    Journal of Bacteriology & Mycology: Open Access Research Article Open Access Black grain eumycetoma revisited: enhanced fungal isolation by use of a novel technique Abstract Volume 4 Issue 5 - 2017 Background: Dematiaceous fungi or brown-black fungi distributed worldwide are being Hazra N,1 Gupta RM,2 Verma R,3 Gera V3 increasingly recognized as pathogens of importance. Found in soil, they have unique 1Depatment of Laboratory Medicine, Command Hospital pathogenic mechanisms owing to the presence of melanin in their cell walls, which Central Command, India imparts the characteristic dark color to their spores and hyphae. Eumycetoma is a common, 2Department of Lab Sciences and Molecular Medicine, Army chronic, granulomatous infection that is present worldwide and endemic in tropical and Hospital (R&R), India subtropical regions. Black grain eumycetoma accounts for about 95% of eumycetomas in 3Department of Dermatology, Base Hospital, India India. Isolation of the causative fungus from the coloured grains in the laboratory is an arduous task resulting in empirical treatment. At present, clear evidence based treatment Correspondence: Dr. Col Nandita Hazra, MD(Microbiol), recommendations are unavailable and it becomes pertinent to identify, isolate and speciate Trained in Neurovirology (NIMHANS), PSG FAIMER Fellow the causative fungus so that appropriate treatment can be instituted for complete resolution. 2010.Sr Adv(Path & Micro) & HOD, Department of Laboratory Medicine, Command Hospital (Central Command), Lucknow Objective: This article revisits black grain mycetoma in the larger spectrum of 226002, India, Email [email protected] phaeohyphomycosis, describes four patients with black grain mycetoma and elaborates on a novel technique developed in our laboratory for assured isolation of fungi from eumycotic Received: January 27, 2017 | Published: May 10, 2017 “grains”.
    [Show full text]
  • Madurella Mycetomatis Grain Model in Galleria Mellonella Larvae
    RESEARCH ARTICLE A Madurella mycetomatis Grain Model in Galleria mellonella Larvae Wendy Kloezen1, Marilyn van Helvert-van Poppel2, Ahmed H. Fahal3, Wendy W. J. van de Sande1* 1 Erasmus University Medical Center, Department of Medical Microbiology and Infectious Diseases, Rotterdam, The Netherlands, 2 St. Elisabeth Ziekenhuis, Department of Clinical Pathology, Tilburg, The Netherlands, 3 Mycetoma Research Centre, Soba University, Khartoum, Sudan * [email protected] Abstract Eumycetoma is a chronic granulomatous subcutaneous infectious disease, endemic in tropical and subtropical regions and most commonly caused by the fungus Madurella myce- tomatis. Interestingly, although grain formation is key in mycetoma, its formation process and its susceptibility towards antifungal agents are not well understood. This is because OPEN ACCESS grain formation cannot be induced in vitro; a mammalian host is necessary to induce its for- mation. Until now, invertebrate hosts were never used to study grain formation in M. myce- Citation: Kloezen W, van Helvert-van Poppel M, Fahal AH, van de Sande WWJ (2015) A Madurella tomatis. In this study we determined if larvae of the greater wax moth Galleria mellonella mycetomatis Grain Model in Galleria mellonella could be used to induce grain formation when infected with M. mycetomatis. Three different Larvae. PLoS Negl Trop Dis 9(7): e0003926. M. mycetomatis strains were selected and three different inocula for each strain were used doi:10.1371/journal.pntd.0003926 to infect G. mellonella larvae, ranging from 0.04 mg/larvae to 4 mg/larvae. Larvae were mon- Editor: Bodo Wanke, Fundação Oswaldo Cruz, itored for 10 days. It appeared that most larvae survived the lowest inoculum, but at the BRAZIL highest inoculum all larvae died within the 10 day observation period.
    [Show full text]