Döğen Aylin, Metin Banu, Ilkit Macit, de Hoog G Sybren, Heitman Joseph
Department of Pharmaceutical Microbiology, Faculty of Pharmacy, University of Mersin, Mersin, Turkey.
Department of Food Engineering, Faculty of Engineering and Natural Sciences, Istanbul Sabahattin Zaim University, Istanbul, Turkey.
PLoS One. 2017 Aug 3;12(8):e0182653. doi: 10.1371/journal.pone.0182653. eCollection 2017.
Reference isolates of Candida parapsilosis (n = 8), Candida metapsilosis (n = 6), Candida orthopsilosis (n = 7), and Lodderomyces elongisporus (n = 11) were analyzed to gain insight into their pathobiology and virulence mechanisms. Initial evaluation using BBL Chromagar Candida medium misidentified L. elongisporus isolates as C. albicans. Polymerase chain reaction analysis of isolate MTL idiomorphs revealed that all C. parapsilosis isolates were MTLa homozygous and no MTL α1, α2, a1, or a2 gene was detected in L. elongisporus isolates. For C. orthopsilosis, two isolates were MTLa homozygous and five were MTL-heterozygous. Similarly, one C. metapsilosis isolate was MTLα homozygous whereas five were MTL-heterozygous. Isolate phenotypic switching analysis revealed potential phenotypic switching in the MTLα homozygous C. metapsilosis isolate, resulting in concomitant elongated cell formation. Minimum inhibitory concentrations of fluconazole (FLC) and FK506, alone or in combination, were determined by checkerboard assay, with data analyzed using the fractional inhibitory concentration index model. Synergistic or additive effects of these compounds were commonly observed in C. parapsilosis and L. elongisporus isolates. No killer activity was observed in the studied isolates, as determined phenotypically. No significant difference in virulence was seen for the four species in a Galleria mellonella model (P > 0.05). In conclusion, our results demonstrated phenotypic switching of C. metapsilosis CBS 2315 and that FLC and FK506 represent a promising drug combination against C. parapsilosis and L. elongisporus. The findings of the present study contribute to our understanding of the biology, diagnosis, and new possible treatments of the C. parapsilosis species group and L. elongisporus.
对近平滑念珠菌(n = 8)、间型念珠菌(n = 6)、正平滑念珠菌(n = 7)和长孢洛德酵母(n = 11)的参考菌株进行分析,以深入了解它们的致病生物学和毒力机制。使用BBL Chromagar念珠菌培养基进行的初步评估将长孢洛德酵母菌株误鉴定为白色念珠菌。对分离株MTL特异型进行聚合酶链反应分析表明,所有近平滑念珠菌分离株均为MTLa纯合子,而在长孢洛德酵母分离株中未检测到MTL α1、α2、a1或a2基因。对于正平滑念珠菌,两个分离株为MTLa纯合子,五个为MTL杂合子。同样,一个间型念珠菌分离株为MTLα纯合子,而五个为MTL杂合子。分离株表型转换分析显示,MTLα纯合间型念珠菌分离株存在潜在的表型转换,导致伴随的细胞伸长形成。通过棋盘法测定氟康唑(FLC)和FK506单独或联合使用时的最低抑菌浓度,并使用分数抑菌浓度指数模型分析数据。在近平滑念珠菌和长孢洛德酵母分离株中通常观察到这些化合物的协同或相加作用。在所研究的分离株中,通过表型测定未观察到杀伤活性。在大蜡螟模型中,这四个物种的毒力没有显著差异(P > 0.05)。总之,我们的结果证明了间型念珠菌CBS 2315的表型转换,并且FLC和FK506是针对近平滑念珠菌和长孢洛德酵母的有前景的药物组合。本研究结果有助于我们理解近平滑念珠菌种组和长孢洛德酵母的生物学、诊断和新的可能治疗方法。