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Combatting Body Odor by the Means of Microbial Transplantations

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Combatting Body Odor by the Means of Microbial Transplantations Faculty of Bio-science engineering Academic year 2013 – 2014 Combatting body odor by the means of microbial transplantations Murielle Meunier Promotor: Prof. dr. ir. Tom Van De Wiele Promotor: Prof. dr. ir. Nico Boon Tutor: dr. ir. Chris Callewaert A master thesis submitted in partial fulfillment of the requirements for the degree of Master in Bio-science engineering Copyright / Confidentiality “This document may contain confidential information proprietary to the University of Ghent. It is therefore strictly forbidden to publish, cite or make public in any way this document or any part thereof without the express written permission by the University of Ghent. Under no circumstance this document may be communicated to or put at the disposal of third parties; photocopying or duplicating it in any other way is strictly prohibited. Disregarding the confidential nature of this document may cause irremediable damage to the University of Ghent. The author and the promotors do not give the permission to use this thesis for consultation and to copy parts of it for personal use. Every use is subject to the copyright laws, more specifically the source must be extensively specified when using results from this thesis” Ghent, June 2014 The promotors The author Prof. dr. ir. Tom Van de Wiele Murielle Meunier Prof. dr. ir. Nico Boon Acknowledgements This work has been brought to life due to the support and contribution of many people who I would like to thank in particular. First, special thanks to my tutor ir. Chris Callewaert, for his infinite support and comprehension. My promotors prof. dr. ir. Nico Boon and prof. dr. ir. Tom Van De Wiele have intensively guided me this year and were always keen to offer their professional advice on the topic. I would like to thank the LabMET department for the use of its labs and all its products and in special, some great people within: Tim Lacoere, who trained me in the incredibly complex mechanism of the DGGE system, Frederiek-Maarten, to share his wisdom about statistics, Emma, for her kindness and helpfulness in the lab, and Jeroen, for answering my numberous Excel and Word questions. I wish I could offer a medal to each member of the scent panel, for voluntary enduring all sorts of axillary scents for a whole year. Furthermore, I would like to thank dr. Jo Lambert and Mireille van Gele, of the department of dermatology of Ghent University, because of their contribution in their area of specialization. At last, but perhaps the most important: all my fellow colleagues at LabMET, who were always available for a chat or a laugh to brighten up your day. Abstract Unpleasant body odor, also called bromhidrosis, can lead to very uncomfortable and even embarrassing situations. Until now, this could only be resolved temporarily by the use of soap, deodorants, anti-perspirants, or for a longer time by the application of radical solutions, such as botox-injections or hidrosuction. This research presents a new solution to combat body odor, by the application of microbial transplants. Freshly produced sweat does not lead to any odor. Bacteria, who convert the compounds in sweat, cause body odor. Two important bacterial groups occur in the axillary region: Staphylococcus spp. and Corynebacterium spp. Corynebacterium is known as the main contributor of body odor. Microbial transplantations in combination with antibiotics were applied to reduce Corynebacterium abundances. This treatment led to an enrichment of Staphylococcus species and an improvement of the hedonic values of the body odor. Comparison between the treated group, that receive both antibiotics and microbial transplantations, and the placebo group, that only receive antibiotics, led to a significant higher increase in Staphylococcus in the treatment group (p<0.0001). This study offers good prospects to permanently cure patients who suffer from bromhidrosis and it warrants further investigation on long term effects of microbial axillary transplantations. Keywords: armpit, body odor, bacterial community, DGGE, microbial transplantation, bacteriotherapy, Staphylococcus spp., Corynebacterium spp. Samenvatting Onaangename lichaamsgeur, in medische termen gekend als bromhidrosis, kan leiden tot zeer onaangename en soms zelfs genante situaties. Tot vandaag kon dit enkel tijdelijk bestreden worden met deodoranten, antiperspiranten, etc. Voor een permanente oplossing moest men zich wenden tot radicale oplossingen zoals botox injecties of hidrosuctie. Dit onderzoek stelt een nieuwe manier voor om dit probleem permanent op te lossen, door middel van microbiële transplantaties. Vers geproduceerd zweet heeft namelijk geen geur. Lichaamsgeur wordt veroorzaakt door de bacteriën die zich op de huid bevinden en het zweet metaboliseren. Twee belangrijke bacteriële groepen komen voor in de okselregio: Staphylococcus spp. en Corynebacterium spp. Corynebacterium is gekend als de belangrijkste bijdrager van okselgeur. Microbiële transplantaties, in combinatie met antibiotica, werden aangebracht om Corynebacterium abundanties te verlagen. Deze behandeling heeft geleid tot een aanrijking van Staphylococcus species en een verbetering van de lichaamsgeur. De vergelijking tussen de behandelde groep, die zowel antibiotica als een microbiële transplantatie kreeg, en de placebo groep, die enkel antibiotica kreeg, leidde tot een significante aanrijking in Staphylococcus in de behandelde groep (p<0.0001). Deze studie biedt een goede basis om bromhidrosis op een eenvoudige manier permanent op te lossen en duidt op het belang om de effecten van microbiële transplantaties op lange termijn verder te onderzoeken. Kernwoorden: oksel, okselgeur, bacteriële gemeenschap, DGGE, microbiële transplantatie, bacteriotherapie, Staphylococcus spp., Corynebacterium spp. Table of contents List of abbreviations ........................................................................................................ 0 1. Introduction .............................................................................................................. 1 2. Literature study ........................................................................................................ 2 2.3. The skin ............................................................................................................. 2 2.4. Sweat ................................................................................................................. 6 2.5. Bacteria ............................................................................................................. 6 2.6. The origin of sweat odor ................................................................................. 14 2.7. Microbial transplantations ............................................................................... 20 3. Research question ............................................................................................... 20 4. Materials and methods .......................................................................................... 22 4.3. Pre-investigation ............................................................................................. 22 4.4. Selection of test subjects and donors ............................................................. 23 4.5. First series ....................................................................................................... 23 4.6. Second series .................................................................................................. 25 4.7. DNA-analysis ................................................................................................... 26 4.7.1. DNA-extraction ......................................................................................... 26 4.7.2. PCR .......................................................................................................... 27 4.7.3. Gel electrophoresis (GE) .......................................................................... 28 4.7.4. Denaturing gradient gel electrophoresis (DGGE) ................................... 29 4.8. Scent panel...................................................................................................... 31 4.9. Self-evaluation ................................................................................................. 32 4.10. Statistical analysis ....................................................................................... 32 Pre-investigation .................................................................................................... 32 Treatment .............................................................................................................. 32 Effect of antibiotics ................................................................................................ 33 5. Results ................................................................................................................... 34 5.3. Pre-investigation ............................................................................................. 34 5.4. Nested PCR .................................................................................................... 37 5.5. Treatment series one ...................................................................................... 38 5.6. Second series .................................................................................................. 44 5.6.1. Treatment ................................................................................................. 44 5.6.2. Placebo treatment ...................................................................................
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