Postgraduate Program in Medical Microbiology, Specialized Medical Mycology Center, Federal University of Ceará, Rua Coronel Nunes de Melo, s/n, Rodolfo Te ófi lo. CEP: 60430270, Fortaleza-CE, Brazil.
Med Mycol. 2013 Jul;51(5):555-60. doi: 10.3109/13693786.2012.752878. Epub 2013 Jan 4.
This study aimed at evaluating the in vitro antifungal susceptibility of Candida albicans isolates obtained during necropsy of a wild Brazilian porcupine and the mechanism of azole resistance. Initially, we investigated the in vitro susceptibility of the three isolates to amphotericin B, caspofungin, fluconazole, itraconazole, ketoconazole and voriconazole. Afterwards, three sub-inhibitory concentrations (47, 21 and 12 mg/l) of promethazine, an efflux pump inhibitor, were tested in combination with the antifungal drugs in order to evaluate the role of these pumps in the development of antifungal resistance. In addition, the three isolates were submitted to RAPD-PCR and M13-fingerprinting analyses. The minimum inhibitory concentrations (MICs) obtained with the isolates were 1, 0.03125, 250, 125, 8 and 250 mg/l for amphotericin B, caspofungin, fluconazole, itraconazole, ketoconazole and voriconazole, respectively, and the isolates were found to be resistant to all tested azoles. The addition of the three subinhibitory concentrations of promethazine resulted in statistically significant (P < 0.05) reductions in the MICs for all tested drugs, with decreases to azoles being statistically greater than those for amphotericin B and caspofungin (P < 0.05). The molecular analyses showed a genetic similarity among the three tested isolates, suggesting the occurrence of candidemia in the studied animal. These findings highlight the importance of monitoring antifungal susceptibility of Candida spp. from veterinary sources, especially as they may indicate the occurrence of primary azole resistance even in wild animals.
本研究旨在评估从一只巴西野生豪猪尸检中获得的白色念珠菌分离株的体外抗真菌药敏性及其唑类耐药机制。首先,我们研究了这三种分离株对两性霉素 B、卡泊芬净、氟康唑、伊曲康唑、酮康唑和伏立康唑的体外药敏性。随后,我们测试了三种亚抑制浓度(47、21 和 12 mg/L)的吩噻嗪,一种外排泵抑制剂,与抗真菌药物联合使用,以评估这些泵在抗真菌药物耐药性发展中的作用。此外,还对这三种分离株进行了 RAPD-PCR 和 M13 指纹图谱分析。三种分离株的最低抑菌浓度(MIC)分别为两性霉素 B(1 mg/L)、卡泊芬净(0.03125 mg/L)、氟康唑(250 mg/L)、伊曲康唑(125 mg/L)、酮康唑(8 mg/L)和伏立康唑(250 mg/L),且三种分离株均对所有测试的唑类药物耐药。三种亚抑制浓度的吩噻嗪的加入使所有测试药物的 MIC 值均有统计学意义(P<0.05)的降低,且与两性霉素 B 和卡泊芬净相比,降低唑类药物的 MIC 值的效果更显著(P<0.05)。分子分析表明,三种测试分离株之间存在遗传相似性,提示研究动物发生了念珠菌血症。这些发现强调了监测兽医来源的念珠菌属抗真菌药敏性的重要性,特别是因为它们可能表明即使在野生动物中也存在原发性唑类耐药的发生。