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皮肤癣菌体外和体内生物膜形成能力的定量与结构分析

Quantitative and structural analyses of the in vitro and ex vivo biofilm-forming ability of dermatophytes.

作者信息

Brilhante Raimunda Sâmia Nogueira, Correia Edmilson Emanuel Monteiro, Guedes Glaucia Morgana de Melo, Pereira Vandbergue Santos, Oliveira Jonathas Sales de, Bandeira Silviane Praciano, Alencar Lucas Pereira de, Andrade Ana Raquel Colares de, Castelo-Branco Débora de Souza Collares Maia, Cordeiro Rossana de Aguiar, Pinheiro Adriana de Queiroz, Chaves Lúcio Jackson Queiroz, Pereira Neto Waldemiro de Aquino, Sidrim José Júlio Costa, Rocha Marcos Fábio Gadelha

机构信息

Department of Pathology and Legal Medicine, School of Medicine, Specialized Medical Mycology Center, Postgraduate Program in Medical Microbiology, Federal University of Ceará, Fortaleza-CE, Brazil.

School of Veterinary, Postgraduate Program in Veterinary Science, State University of Ceará, Fortaleza-CE, Brazil.

出版信息

J Med Microbiol. 2017 Jul;66(7):1045-1052. doi: 10.1099/jmm.0.000528. Epub 2017 Jul 15.

Abstract

PURPOSE

The aim of this study was to evaluate the in vitro and ex vivo biofilm-forming ability of dermatophytes on a nail fragment.

METHODOLOGY

Initially, four isolates of Trichophyton rubrum, six of Trichophyton tonsurans, three of Trichophyton mentagrophytes, ten of Microsporum canis and three of Microsporum gypseum were tested for production biomass by crystal violet assay. Then, one strain per species presenting the best biofilm production was chosen for further studies by optical microscopy (Congo red staining), confocal laser scanning (LIVE/DEAD staining) and scanning electron (secondary electron) microscopy.

RESULTS

Biomass quantification by crystal violet assay, optical microscope images of Congo red staining, confocal microscope and scanning electron microscope images revealed that all species studied are able to form biofilms both in vitro and ex vivo, with variable density and architecture. M. gypseum, T. rubrum and T. tonsurans produced robust biofilms, with abundant matrix and biomass, while M. canis produced the weakest biofilms compared to other species.

CONCLUSION

This study sheds light on biofilms of different dermatophyte species, which will contribute to a better understanding of the pathophysiology of dermatophytosis. Further studies of this type are necessary to investigate the processes involved in the formation and composition of dermatophyte biofilms.

摘要

目的

本研究旨在评估皮肤癣菌在指甲碎片上的体外和离体生物膜形成能力。

方法

首先,通过结晶紫测定法对4株红色毛癣菌、6株断发毛癣菌、3株须癣毛癣菌、10株犬小孢子菌和3株石膏样小孢子菌进行生物量产生测试。然后,每种菌选择一株生物膜产生最佳的菌株,通过光学显微镜(刚果红染色)、共聚焦激光扫描显微镜(活/死染色)和扫描电子显微镜(二次电子)进行进一步研究。

结果

通过结晶紫测定法进行的生物量定量、刚果红染色的光学显微镜图像、共聚焦显微镜和扫描电子显微镜图像显示,所有研究的菌种都能够在体外和离体形成生物膜,密度和结构各不相同。石膏样小孢子菌、红色毛癣菌和断发毛癣菌产生强大的生物膜,具有丰富的基质和生物量,而犬小孢子菌与其他菌种相比产生的生物膜最弱。

结论

本研究揭示了不同皮肤癣菌物种的生物膜情况,这将有助于更好地理解皮肤癣菌病的病理生理学。有必要进行此类进一步研究,以调查皮肤癣菌生物膜形成和组成所涉及的过程。

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