Characterizing clinical isolates of Acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in Taiwan

Background/Purpose: Acanthamoeba keratitis (AK), a painful infectious corneal disease, is caused by the free-living pathogenic species Acanthamoeba. The symptoms include corneal infiltrate, epithelial, and stromal destruction, and loss of vision. Current treatment generally involves an hourly applic...

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Main Authors: Fu-Chin Huang, Min-Hsiu Shih, Kai-Fei Chang, Jian-Ming Huang, Jyh-Wei Shin, Wei-Chen Lin
Format: Article
Language:English
Published: Elsevier 2017-10-01
Series:Journal of Microbiology, Immunology and Infection
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S1684118215009081
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author Fu-Chin Huang
Min-Hsiu Shih
Kai-Fei Chang
Jian-Ming Huang
Jyh-Wei Shin
Wei-Chen Lin
author_facet Fu-Chin Huang
Min-Hsiu Shih
Kai-Fei Chang
Jian-Ming Huang
Jyh-Wei Shin
Wei-Chen Lin
author_sort Fu-Chin Huang
collection DOAJ
description Background/Purpose: Acanthamoeba keratitis (AK), a painful infectious corneal disease, is caused by the free-living pathogenic species Acanthamoeba. The symptoms include corneal infiltrate, epithelial, and stromal destruction, and loss of vision. Current treatment generally involves an hourly application of polyhexamethylene biguanide (PHMB) over a period of several days; however, even this is not entirely effective against all strains/isolates. The aims of this study were to confirm the existence of pathogenic strains in Taiwan which are highly resistant to drugs and to characterize the behavior of these strains. Methods: An in vitro Acanthamoeba species culture platform was established to observe the effectiveness of treatment and chart the morphological changes that occur under the effects of drugs using a light microscope and time-lapse recording. Changes in gene expression were examined using reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR. Results: Over 90% of the standard strain cells (ATCC 30010) were lysed after being treated with PHMB for 1 hour; however, clinical isolates of Acanthamoeba castellanii that differed in their susceptibility to the treatment drug were only partly lysed. Following treatment with PHMB, National Cheng Kung University Hospital isolation B (NCKH_B) transformed into a pseudocyst under the effects of drug stress; however, National Cheng Kung University Hospital isolation D (NCKH_D), an isolate with higher tolerance for PHMB, did not transform. Conclusion: Our results confirm the existence of clinical isolates of A. castellanii with high resistance to PHMB in Taiwan and present the alternative drug tolerance of A. castellanii in addition to the transformation of pseudocyst/cyst.
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spelling doaj.art-24b0c32a6e834a7aba859482aecd16f42022-12-22T00:49:04ZengElsevierJournal of Microbiology, Immunology and Infection1684-11822017-10-0150557057710.1016/j.jmii.2015.10.011Characterizing clinical isolates of Acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in TaiwanFu-Chin Huang0Min-Hsiu Shih1Kai-Fei Chang2Jian-Ming Huang3Jyh-Wei Shin4Wei-Chen Lin5Department of Ophthalmology, National Cheng Kung University Hospital, Tainan, TaiwanDepartment of Ophthalmology, National Cheng Kung University Hospital, Tainan, TaiwanDepartment of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, TaiwanDepartment of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, TaiwanDepartment of Parasitology, National Cheng Kung University, Tainan, TaiwanDepartment of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, TaiwanBackground/Purpose: Acanthamoeba keratitis (AK), a painful infectious corneal disease, is caused by the free-living pathogenic species Acanthamoeba. The symptoms include corneal infiltrate, epithelial, and stromal destruction, and loss of vision. Current treatment generally involves an hourly application of polyhexamethylene biguanide (PHMB) over a period of several days; however, even this is not entirely effective against all strains/isolates. The aims of this study were to confirm the existence of pathogenic strains in Taiwan which are highly resistant to drugs and to characterize the behavior of these strains. Methods: An in vitro Acanthamoeba species culture platform was established to observe the effectiveness of treatment and chart the morphological changes that occur under the effects of drugs using a light microscope and time-lapse recording. Changes in gene expression were examined using reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR. Results: Over 90% of the standard strain cells (ATCC 30010) were lysed after being treated with PHMB for 1 hour; however, clinical isolates of Acanthamoeba castellanii that differed in their susceptibility to the treatment drug were only partly lysed. Following treatment with PHMB, National Cheng Kung University Hospital isolation B (NCKH_B) transformed into a pseudocyst under the effects of drug stress; however, National Cheng Kung University Hospital isolation D (NCKH_D), an isolate with higher tolerance for PHMB, did not transform. Conclusion: Our results confirm the existence of clinical isolates of A. castellanii with high resistance to PHMB in Taiwan and present the alternative drug tolerance of A. castellanii in addition to the transformation of pseudocyst/cyst.http://www.sciencedirect.com/science/article/pii/S1684118215009081Acanthamoeba keratitisdrug tolerancepolyhexamethylene biguanide
spellingShingle Fu-Chin Huang
Min-Hsiu Shih
Kai-Fei Chang
Jian-Ming Huang
Jyh-Wei Shin
Wei-Chen Lin
Characterizing clinical isolates of Acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in Taiwan
Journal of Microbiology, Immunology and Infection
Acanthamoeba keratitis
drug tolerance
polyhexamethylene biguanide
title Characterizing clinical isolates of Acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in Taiwan
title_full Characterizing clinical isolates of Acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in Taiwan
title_fullStr Characterizing clinical isolates of Acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in Taiwan
title_full_unstemmed Characterizing clinical isolates of Acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in Taiwan
title_short Characterizing clinical isolates of Acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in Taiwan
title_sort characterizing clinical isolates of acanthamoeba castellanii with high resistance to polyhexamethylene biguanide in taiwan
topic Acanthamoeba keratitis
drug tolerance
polyhexamethylene biguanide
url http://www.sciencedirect.com/science/article/pii/S1684118215009081
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