Mello Thaís Pereira de, Aor Ana Carolina, Gonçalves Diego de Souza, Seabra Sergio Henrique, Branquinha Marta Helena, Santos André Luis Souza Dos
Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brasil.
Laboratório de Tecnologia em Cultura de Células, Centro Universitário Estadual da Zona Oeste, Rio de Janeiro, RJ, Brasil.
Mem Inst Oswaldo Cruz. 2018;113(6):e180102. doi: 10.1590/0074-02760180102. Epub 2018 Jun 18.
BACKGROUND Scedosporium/Lomentospora species are opportunistic mould pathogens, presenting notable antifungal resistance. OBJECTIVES/METHODS We analysed the conidia and germinated conidia of S. apiospermum (Sap), S. aurantiacum (Sau), S. minutisporum (Smi) and L. prolificans (Lpr) by scanning electron microscopy and exposition of surface molecules by fluorescence microscopy. FINDINGS Conidia of Sap, Smi and Sau had oval, ellipsoidal and cylindrical shape, respectively, with several irregularities surrounding all surface areas, whereas Lpr conidia were rounded with a smooth surface. The germination of Sap occurred at the conidial bottom, while Smi and Sau germination primarily occurred at the centre of the conidial cell, and Lpr germination initiated at any part of the conidial surface. The staining of N-acetylglucosamine-containing molecules by fluorescein-labelled WGA primarily occurred during the germination of all studied fungi and in the conidial scars, which is the primary location of germination. Calcofluor white, which recognises the polysaccharide chitin, strongly stained the conidial cells and, to a lesser extent, the germination. Both mannose-rich glycoconjugates (evidenced by fluoresceinated-ConA) and cell wall externally located polypeptides presented distinct surface locations and expression according to both morphotypes and fungal species. In contrast, sialic acid and galactose-containing structures were not detected at fungal surfaces. MAIN CONCLUSIONS The present study demonstrated the differential production/exposition of surface molecules on distinct morphotypes of Scedosporium/Lomentospora species.
帚霉属/罗门孢属真菌是机会性霉菌病原体,具有显著的抗真菌耐药性。
目的/方法:我们通过扫描电子显微镜分析了阿皮奥斯波氏帚霉(Sap)、橙色帚霉(Sau)、微小帚霉(Smi)和多育罗门孢(Lpr)的分生孢子和已萌发的分生孢子,并通过荧光显微镜观察表面分子的暴露情况。
Sap、Smi和Sau的分生孢子分别呈椭圆形、椭球形和圆柱形,所有表面区域都有一些不规则之处,而Lpr的分生孢子呈圆形,表面光滑。Sap的萌发发生在分生孢子底部,而Smi和Sau的萌发主要发生在分生孢子细胞的中心,Lpr的萌发则始于分生孢子表面的任何部位。用荧光素标记的麦胚凝集素(WGA)对含N-乙酰葡糖胺分子进行染色,主要发生在所有研究真菌的萌发过程中以及分生孢子痕处,分生孢子痕是萌发的主要部位。识别多糖几丁质的荧光增白剂强烈地染色了分生孢子细胞,对萌发的染色程度较轻。富含甘露糖的糖缀合物(由荧光素化刀豆球蛋白A证明)和细胞壁外部定位的多肽根据形态类型和真菌种类呈现出不同的表面位置和表达情况。相比之下,在真菌表面未检测到含唾液酸和半乳糖的结构。
本研究证明了帚霉属/罗门孢属真菌不同形态类型表面分子的差异产生/暴露情况。