A protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans

Abstract Contaminations and fastidiousness of M. ulcerans may have both hamper isolation of strains from environmental sources. We aimed to optimize decontamination and culture of environmental samples to circumvent both limitations. Three strains of M. ulcerans cultured onto Middlebrook 7H10 at 30 ...

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Autores principales:	Dezemon Zingue, Arup Panda, Michel Drancourt
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3126b6ec14b24096a8c9bd8ddc3c9579
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spelling	oai:doaj.org-article:3126b6ec14b24096a8c9bd8ddc3c95792021-12-02T16:08:15ZA protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans10.1038/s41598-018-25278-y2045-2322https://doaj.org/article/3126b6ec14b24096a8c9bd8ddc3c95792018-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-25278-yhttps://doaj.org/toc/2045-2322Abstract Contaminations and fastidiousness of M. ulcerans may have both hamper isolation of strains from environmental sources. We aimed to optimize decontamination and culture of environmental samples to circumvent both limitations. Three strains of M. ulcerans cultured onto Middlebrook 7H10 at 30 °C for 20 days yielded a significantly higher number of colonies in micro-aerophilic atmosphere compared to ambient atmosphere, 5% CO2 and anaerobic atmosphere. In a second step, we observed that M. ulcerans genome uniquely encoded chitinase, fucosidase and A-D-GlcNAc-diphosphoryl polyprenol A-3-L-rhamnosyl transferase giving M. ulcerans the potential to metabolize chitine, fucose and N-acetyl galactosamine (NAG), respectively. A significant growth-promoting effect of 0.2 mg/mL chitin (p < 0.05), 0.01 mg/mL N-acetyl galactosamine (p < 0.05), 0.01 mg/mL fucose (p < 0.05) was observed with M. ulcerans indicating that NAG alone or combined with fucose and chitin could complement Middlebrook 7H10. Finally, the protocol combining 1% chlorhexidine decontamination with micro-aerophilic incubation on Middlebrook 7H10 medium containing chitin (0.2%), NAG (0.01%) and fucose (0.01%) medium and auto-fluorescence detection of colonies allowed for the isolation of one mycolactone-encoding strain from Thryonomys swinderianus (aulacode) feces specimens collected near the Kossou Dam, Côte d’Ivoire. We propose that incubation of chlorhexidine-decontaminated environmental specimens on Middlebrook 7H10-enriched medium under micro-aerophilic atmosphere at 30 °C may be used for the tentative isolation of M. ulcerans strains from potential environmental sources.Dezemon ZingueArup PandaMichel DrancourtNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-12 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Dezemon Zingue Arup Panda Michel Drancourt A protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans
description	Abstract Contaminations and fastidiousness of M. ulcerans may have both hamper isolation of strains from environmental sources. We aimed to optimize decontamination and culture of environmental samples to circumvent both limitations. Three strains of M. ulcerans cultured onto Middlebrook 7H10 at 30 °C for 20 days yielded a significantly higher number of colonies in micro-aerophilic atmosphere compared to ambient atmosphere, 5% CO2 and anaerobic atmosphere. In a second step, we observed that M. ulcerans genome uniquely encoded chitinase, fucosidase and A-D-GlcNAc-diphosphoryl polyprenol A-3-L-rhamnosyl transferase giving M. ulcerans the potential to metabolize chitine, fucose and N-acetyl galactosamine (NAG), respectively. A significant growth-promoting effect of 0.2 mg/mL chitin (p < 0.05), 0.01 mg/mL N-acetyl galactosamine (p < 0.05), 0.01 mg/mL fucose (p < 0.05) was observed with M. ulcerans indicating that NAG alone or combined with fucose and chitin could complement Middlebrook 7H10. Finally, the protocol combining 1% chlorhexidine decontamination with micro-aerophilic incubation on Middlebrook 7H10 medium containing chitin (0.2%), NAG (0.01%) and fucose (0.01%) medium and auto-fluorescence detection of colonies allowed for the isolation of one mycolactone-encoding strain from Thryonomys swinderianus (aulacode) feces specimens collected near the Kossou Dam, Côte d’Ivoire. We propose that incubation of chlorhexidine-decontaminated environmental specimens on Middlebrook 7H10-enriched medium under micro-aerophilic atmosphere at 30 °C may be used for the tentative isolation of M. ulcerans strains from potential environmental sources.
format	article
author	Dezemon Zingue Arup Panda Michel Drancourt
author_facet	Dezemon Zingue Arup Panda Michel Drancourt
author_sort	Dezemon Zingue
title	A protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans
title_short	A protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans
title_full	A protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans
title_fullStr	A protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans
title_full_unstemmed	A protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans
title_sort	protocol for culturing environmental strains of the buruli ulcer agent, mycobacterium ulcerans
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/3126b6ec14b24096a8c9bd8ddc3c9579
work_keys_str_mv	AT dezemonzingue aprotocolforculturingenvironmentalstrainsoftheburuliulceragentmycobacteriumulcerans AT aruppanda aprotocolforculturingenvironmentalstrainsoftheburuliulceragentmycobacteriumulcerans AT micheldrancourt aprotocolforculturingenvironmentalstrainsoftheburuliulceragentmycobacteriumulcerans AT dezemonzingue protocolforculturingenvironmentalstrainsoftheburuliulceragentmycobacteriumulcerans AT aruppanda protocolforculturingenvironmentalstrainsoftheburuliulceragentmycobacteriumulcerans AT micheldrancourt protocolforculturingenvironmentalstrainsoftheburuliulceragentmycobacteriumulcerans
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A protocol for culturing environmental strains of the Buruli ulcer agent, Mycobacterium ulcerans

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