Öner Sedef Zeliha, Küçükakın Yaka Saniye, Akçaoğlu Tahsin, Vural Caner, Yılmaz Uğur, Ergin Çağrı
Pamukkale University Faculty of Medicine, Department of Medical Microbiology, Denizli, Türkiye.
Pamukkale University Faculty of Medicine, Department of Ophtalmology, Denizli, Türkiye.
Mikrobiyol Bul. 2023 Oct;57(4):690-697. doi: 10.5578/mb.20239957.
Fungal keratitis is a medical emergency that is among the most common causes of blindness in developing countries. The type of the agent may vary depending on the geographical conditions under which the patient lives, trauma exposure, the use of contact lenses and profession. Curvularia spp. is a saprophytic genus that rarely causes systemic disease in humans and has 250 species identified to date. They proliferate in soil and plants and spread to the environment with their spores and the formation of blackish and fluffy colonies is its most well-known morphological feature. There may be difficulties in cultivating brown (dematiaceous) fungi. Due to the similarity between the genera, conventional methods remain inadequate for diagnosis. In this report, a case of fungal keratitis associated with C.lunata was presented. Seventy-five years-old female patient admitted to the hospital with the symptoms of stinging pain, blurred vision, and swelling in the right eye. Her symptoms had begun four days ago after her eye was hit by a plant. The patient who had a history of peripheral neuropathy due to diabetes mellitus (DM) was hospitalized with a preliminary diagnosis of keratitis, and in the cultures of the patient's corneal scraping samples, the filamentous, black pigment-forming colonies of the pathogen growing on 5% sheep blood agar and potato dextrose agar showing an aerial hyphal structure, were stained with lactophenol cotton blue and examined under the microscope. The microscopic examination revealed geniculate conidiophores with brown pigmentation. On top of these structures were tetralocular macroconidia, one of which appeared to be larger than the main axis. The fungus was subjected to molecular identification with the prediagnosis of Curvularia/Bipolaris. DNA extraction of the ITS region polymerase chain reaction amplification and Sanger sequencing were performed for molecular identification. Sanger sequencing identified the agent to be Curvularia lunata with a similarity rate of 99.79% (NCBI-GenBank Nucleotide ID: OR365075). In vitro antifungal susceptibility of C.lunata was evaluated by microdilution method. Itraconazole and amphotericin B showed higher activity against C.lunata compared to other antifungals while fluconazole was the least active antifungal. Intrastromal and subconjunctival voriconazole injection was applied to the patient who was unresponsive to empirically initiated oral moxifloxacin and different topical treatments (vancomycin, ceftazidime, flucanozole, ganciclovir, cyclopentolate hydrochloride, hyaluronic acid and trehalose). After injection, right penetrating keratoplasty was applied due to increased thinning of the ulcerated area. No pathogen was detected in cultures taken after keratoplasty. Rare fungi should be considered in cases of keratitis that are difficult to treat. Fungal keratitis caused by brown fungi are clinically similar to each other and effective treatment protocols cannot be implemented without a species identification. Identification of the pathogen will enable genus-specific treatment. This will also help prevent complications that may occur. This article aims to present a case of fungal keratitis associated with C.lunata.
真菌性角膜炎是一种医疗急症,是发展中国家最常见的致盲原因之一。病原体的类型可能因患者居住的地理条件、外伤暴露情况、隐形眼镜的使用以及职业而有所不同。弯孢属是一种腐生菌属,很少在人类中引起全身性疾病,迄今为止已鉴定出250个物种。它们在土壤和植物中繁殖,并通过孢子传播到环境中,形成黑色蓬松菌落是其最著名的形态特征。培养棕色(暗色)真菌可能存在困难。由于各属之间的相似性,传统方法在诊断方面仍然不足。在本报告中,介绍了一例与新月弯孢菌相关的真菌性角膜炎病例。一名75岁女性患者因右眼刺痛、视力模糊和肿胀症状入院。她的症状在四天前眼睛被植物击中后开始出现。该患者有因糖尿病(DM)导致的周围神经病变病史,初步诊断为角膜炎后住院。在患者角膜刮片样本的培养物中,在5%羊血琼脂和马铃薯葡萄糖琼脂上生长的病原体形成丝状、黑色色素的菌落,呈现气生菌丝结构,用乳酚棉蓝染色并在显微镜下检查。显微镜检查发现有棕色色素沉着的膝状分生孢子梗。在这些结构之上是四隔大分生孢子,其中一个似乎比主轴大。对该真菌进行了弯孢属/双极霉属的预诊断分子鉴定。对ITS区域进行DNA提取、聚合酶链反应扩增和桑格测序以进行分子鉴定。桑格测序鉴定病原体为新月弯孢菌,相似度为99.79%(NCBI - GenBank核苷酸ID:OR365075)。通过微量稀释法评估了新月弯孢菌的体外抗真菌药敏性。与其他抗真菌药物相比,伊曲康唑和两性霉素B对新月弯孢菌显示出更高的活性,而氟康唑是活性最低的抗真菌药物。对经验性开始使用口服莫西沙星和不同局部治疗(万古霉素、头孢他啶、氟康唑、更昔洛韦、盐酸环喷托酯、透明质酸和海藻糖)无反应的患者,进行了基质内和结膜下伏立康唑注射。注射后,由于溃疡区域变薄加剧,进行了右穿透性角膜移植术。角膜移植术后采集的培养物中未检测到病原体。对于难以治疗的角膜炎病例,应考虑罕见真菌。由棕色真菌引起的真菌性角膜炎在临床上彼此相似,未经菌种鉴定就无法实施有效的治疗方案。病原体的鉴定将实现属特异性治疗。这也将有助于预防可能发生的并发症。本文旨在介绍一例与新月弯孢菌相关的真菌性角膜炎病例。