Parasites, Hosts and Diseases

"keratitis"

Brief Communication

Molecular detection and characterization of Acanthamoeba infection in dogs and its association with keratitis in Korea: Subin Lee, Badriah Alkathiri, Ji Seung Jung, Nanyoung Kang, Jiyi Hwang, Sang-Eun Park, Yeonchul Hong, Kyung-Mee Park, Seung-Hun Lee; Parasites Hosts Dis 2024;62(1):139-144.
Published online February 23, 2024; DOI: https://doi.org/10.3347/PHD.23112

Abstract
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Acanthamoeba infection is associated with keratitis in humans; however, its association with keratitis in dogs remains unclear. To investigate this possibility, we collected 171 conjunctival swab samples from dogs with eye-related diseases (65 with keratitis and 106 without keratitis) at Chungbuk National University Veterinary Teaching Hospital, Korea, from August 2021 to September 2022. Polymerase chain reaction identified 9 samples (5.3%) as Acanthamoeba positive; of these, 3 were from dogs with keratitis (4.6%) and 6 were from dogs without keratitis (5.7%). Our results indicated no significant association between Acanthamoeba infection and keratitis, season, sex, or age. All Acanthamoeba organisms found in this study had the genotype T4, according to 18S ribosomal RNA analysis. Acanthamoeba infection in dogs might have only a limited association with keratitis.

Citations

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Epidemiology, molecular characterization, and risk factors of Acanthamoeba spp., Blastocystis spp., and Cyclospora spp. infections in snakes in China
Yilei Zhang, Zhouchun Li, Xinyuan Wang, Kaili Gao, Lijie Tian, Olalekan Opeyemi Ayanniyi, Qianming Xu, Congshan Yang
Veterinary Parasitology.2025; 335: 110420. CrossRef
Acanthamoeba spp.: Neglected Protists in Veterinary Medicine
Somayeh Bahrami, Mehdi Zarei, Fiona L. Henriquez, Chukwunonso O. Nzelu
Acta Parasitologica.2025;[Epub] CrossRef
Anti-adherent effects of Rhizophora apiculata bark and leaf extracts and computational prediction of the effects of its compound on β-tubulin interaction in Acanthamoeba triangularis genotype 4
Siriphorn Chimplee, Imran Sama-ae, Suthinee Sangkanu, Watcharapong Mitsuwan, Julalak Chuprom, Rachasak Boonhok, Dhrubo Ahmed Khan, Partha Biswas, Md Nazmul Hasan, Hazel Anne Tabo, Cristina C. Salibay, Polrat Wilairatana, Maria L. Pereira, Muhammad Nawaz,
Veterinary World.2024; : 2829. CrossRef

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Case Report

Demodex Blepharitis: An Analysis of Nine Patients: Young Il Lee, Min Seo, Kyong Jin Cho; Korean J Parasitol 2022;60(6):429-432.
Published online December 22, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.6.429

Abstract
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Inflammatory eyelid symptoms are common in primary care and there have been several reports on Demodex blepharitis. In the present study, we evaluate the 9 patients with Demodex blepharitis, who showed inflammation of the eyelids, dry eye, and cylindrical dandruff at the base of the eyelashes. The causative species from all patients was Demodex folliculorum of either the adult or nymph stage. Two patients had recurrent chalazion and 3 patients had keratitis. Weekly lid scrubs with 50% tee tree oil were performed for 6 weeks. After treatment, the symptoms of blepharitis and keratitis had improved in all patients. This case report provides clinical reference source for the proper treatment of ocular demodicosis.

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Natural Products as Mite Control Agents in Animals: A Review
Fei Liao, Changquan Han, Qingsheng Deng, Ziyao Zhou, Taotao Bao, Menghuai Zhong, Guangyao Tao, Renjun Li, Bo Han, Yanlong Qiao, Yanchun Hu
Molecules.2023; 28(19): 6818. CrossRef

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Brief Communication

Specific Detection of Acanthamoeba species using Polyclonal Peptide Antibody Targeting the Periplasmic Binding Protein of A. castellanii: Min-Jeong Kim, Fu-Shi Quan, Hyun-Hee Kong, Jong-Hyun Kim, Eun-Kyung Moon; Korean J Parasitol 2022;60(2):143-147.
Published online April 20, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.2.143

Abstract
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Acanthamoeba keratitis (AK) is a rare ocular disease, but it is a painful and sight-threatening infectious disease. Early diagnosis and adequate treatment are necessary to prevent serious complications. While AK is frequently diagnosis via several PCR assays or Acanthamoeba-specific antibodies, a more specific and effective diagnostic method is required. This study described the production of a polyclonal peptide antibody against the periplasmic binding protein (PBP) of A. castellanii and investigated its diagnostic potential. Western blot analysis showed that the PBP antibody specifically reacted with the cell lysates of A. castellanii. However, the PBP antibody did not interact with human corneal epithelial (HCE) cells and the other 3 major causative agents of keratitis. Immunocytochemistry (ICC) results revealed the specific detection of A. castellanii trophozoites and cysts by PBP antibodies when A. castellanii were co-cultured with HCE cells. PBP antibody specificity was further confirmed by co-culture of A. castellanii trophozoites with F. solani, S. aureus, and P. aeruginosa via ICC. The PBP antibody specifically reacted with the trophozoites and cysts of A. polyphaga, A. hatchetti, A. culbertsoni, A. royreba, and A. healyi, thus demonstrated its genus-specific nature. These results showed that the PBP polyclonal peptide antibody of A. castellanii could specifically detect several species of Acanthamoeba, contributing to the development of an effective antibody-based AK diagnostics.

Citations

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Evaluation of the potential for diagnosis of fungal keratitis using a Fusarium-specific antibody
Hye-Jeong Jo, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
Scientific Reports.2025;[Epub] CrossRef
Detection of Fusarium solani using cutinase antibody and its application in diagnosing fungal keratitis in an animal model
Hye-Jeong Jo, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon, Olaf Kniemeyer
PLOS One.2025; 20(8): e0330455. CrossRef
A chorismate mutase-targeted, core-shell nanoassembly-activated SERS immunoassay platform for rapid non-invasive detection of Acanthamoeba infection
Hyerin Lee, Min-Jeong Kim, Junkyu Chung, Wansun Kim, Hye-Jeong Jo, Tae Gi Kim, Jae-Ho Shin, Gi-Ja Lee, Fu-Shi Quan, Hyun-Hee Kong, Sang Woong Moon, Eun-Kyung Moon, Samjin Choi
Nano Today.2024; 59: 102506. CrossRef
Evaluating the Diagnostic Potential of Chorismate Mutase Poly-Clonal Peptide Antibody for the Acanthamoeba Keratitis in an Animal Model
Min-Jeong Kim, Hye-Jeong Jo, Hae-Jin Sohn, Ho-Joon Shin, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
Pathogens.2023; 12(4): 526. CrossRef
Detection of Acanthamoeba from Acanthamoeba Keratitis Mouse Model Using Acanthamoeba-Specific Antibodies
Min-Jeong Kim, A-Jeong Ham, A-Young Park, Hae-Jin Sohn, Ho-Joon Shin, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
Microorganisms.2022; 10(9): 1711. CrossRef

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Case Report

The First Acanthamoeba keratitis Case of Non-Contact Lens Wearer with HIV Infection in Thailand: Napaporn Tananuvat, Natnaree Techajongjintana, Pradya Somboon, Anchalee Wannasan; Korean J Parasitol 2019;57(5):505-511.
Published online October 31, 2019; DOI: https://doi.org/10.3347/kjp.2019.57.5.505

Abstract
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Acanthamoeba keratitis (AK) is a rare sight-threatening corneal infection, often reporting from contact lens wearers. An asymptomatic human immunodeficiency virus (HIV)-infected Thai male without history of contact lens use complained foreign body sensation at his left eye during motorbike riding. He had neither specific keratitis symptoms nor common drugs responding, which contributed to delayed diagnosis. By corneal re-scraping, Acanthamoeba-like cysts were detected by calcofluor white staining and agar culture. The etiological agent obtained from the culture was molecularly confirmed by Acanthamoeba spp.-specific PCR, followed by DNA sequencing. The results from BLAST and phylogenetic analysis based on the DNA sequences, revealed that the pathogen was Acanthamoeba T4, the major genotype most frequently reported from clinical isolates. The infection was successfully treated with polyhexamethylene biguanide resulting in corneal scar. This appears the first reported AK case from a non-contact lens wearer with HIV infection in Thailand. Although AK is sporadic in developing countries, a role of free-living Acanthamoeba as an opportunistic pathogen should not be neglected. The report would increase awareness of AK, especially in the case presenting unspecific keratitis symptoms without clinical response to empirical antimicrobial therapy.

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Detection of Free-Living Amoeba in a tertiary care hospital
Nairita Choudhuri, Bhaskar Narayan Chaudhuri, Partha Guchhait, Arup Kumar Dawn, Satadal Das
Microbiologia Medica.2024;[Epub] CrossRef
Diagnosis of Acanthamoeba Keratitis: Past, Present and Future
Matthew Azzopardi, Yu Jeat Chong, Benjamin Ng, Alberto Recchioni, Abison Logeswaran, Darren S. J. Ting
Diagnostics.2023; 13(16): 2655. CrossRef
New Frontiers in Acanthamoeba Keratitis Diagnosis and Management
Omar Shareef, Sana Shareef, Hajirah N. Saeed
Biology.2023; 12(12): 1489. CrossRef
Isolation and identification of potentially pathogenic free-living amoeba in drinking, surface, and stagnant water sources from Alborz Province, Iran
Ehsan Javanmard, Maryam Niyyati, Ali Taghipour, Marziye Fatemi, Hamed Mirjalali, Panagiotis Karanis
Journal of Water and Health.2022; 20(4): 620. CrossRef
Isolation and identification of potentially pathogenic free-living amoeba in dental-unit water samples
Hadi Khatoonaki, Kavous Solhjoo, Hassan Rezanezhad, Belal Armand, Amir Abdoli, Ali Taghipour
Journal of Water and Health.2022; 20(7): 1126. CrossRef
Detection of Acanthamoeba from Acanthamoeba Keratitis Mouse Model Using Acanthamoeba-Specific Antibodies
Min-Jeong Kim, A-Jeong Ham, A-Young Park, Hae-Jin Sohn, Ho-Joon Shin, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
Microorganisms.2022; 10(9): 1711. CrossRef
RETRACTED ARTICLE: Establishment of an Acanthamoeba keratitis mouse model confirmed by amoebic DNA amplification
Heekyoung Kang, Hae-Jin Sohn, A-Young Park, A-Jeong Ham, Jeong-Heon Lee, Young-Hwan Oh, Yong-Joon Chwae, Kyongmin Kim, Sun Park, Hongseok Yang, Suk-Yul Jung, Jong-Hyun Kim, Ho-Joon Shin
Scientific Reports.2021;[Epub] CrossRef
Genotype distribution of Acanthamoeba in keratitis: a systematic review
Maria Luisa Nunes Diehl, Júlia Paes, Marilise Brittes Rott
Parasitology Research.2021; 120(9): 3051. CrossRef
Explicating the presentations of Acanthamoeba keratitis with special concern in the COVID-19 pandemic ambient
Chukwuma Sr Chrysanthus
International Journal of Clinical Microbiology and Biochemical Technology.2021; 4(1): 010. CrossRef
Effectiveness and Safety of Topical Chlorhexidine and Vitamin E TPGS in the Treatment of Acanthamoeba Keratitis: A Survey on 29 Cases
Ciro Caruso, Daniela Eletto, Michele Rinaldi, Luigi Pacente, Salvatore Troisi, Francesco Semeraro, Roberto dell’Omo, Ciro Costagliola
Journal of Clinical Medicine.2020; 9(11): 3775. CrossRef

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Original Articles

Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts: Hae-Jin Sohn, Ga-Eun Seo, Jae-Ho Lee, A-Jeong Ham, Young-Hwan Oh, Heekyoung Kang, Ho-Joon Shin; Korean J Parasitol 2019;57(3):217-223.
Published online June 30, 2019; DOI: https://doi.org/10.3347/kjp.2019.57.3.217

Abstract
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Acanthamoeba castellanii has ubiquitous distribution and causes primary acanthamoebic keratitis (AK). AK is a common disease in contact lens wearers and results in permanent visual impairment or blindness. In this study, we observed the cytopathic effect, in vitro cytotoxicity, and secretion pattern of cytokines in human corneal epithelial cells (HCECs) induced by A. castellanii trophozoites and/or cysts. Morphological observation revealed that panked dendritic HCECs co-cultured with amoeba cysts had changed into round shape and gradually died. Such changes were more severe in co-culture with cyst than those of co-cultivation with trophozoites. In vitro cytotoxicity assay revealed the highest cytotoxicity to HCECs in the co-culture system with amoeba cysts. A. castellanii induced the expression of IL-1α, IL-6, IL-8, and CXCL1 in HCECs. Secreted levels of IL-1α, IL-6, and IL-8 in HCECs co-cultured with both trophozoites and cysts were increased at an early incubation time (3 and 6 hr). These results suggested that cytopathic changes and pro-inflammatory cytokines release of HCECs in response to A. castellanii, especially amoebic cysts, are an important mechanism for AK development.

Citations

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A Synthetic View on Acanthamoeba Keratitis Host Immune Response: Potential Factors Influencing the Development of Chronic Inflammation
Bianca Prado-Costa, Larissa Fagundes Pinto, Mariana Fernandes Fonseca, Denise de Freitas, Larissa Magalhães Alvarenga
Cornea.2025; 44(1): 118. CrossRef
In Vitro Efficacy of Miltefosine Against Clinical Isolates of Acanthamoeba spp. from Patients with Keratitis
Lakshminarayanan Gowtham, Savitri Sharma, Bhupesh Bagga
Seminars in Ophthalmology.2025; 40(8): 767. CrossRef
Diagnostic features of Acanthamoeba keratitis via in vivo confocal microscopy
Joanna Przybek-Skrzypecka, Malcolm Armstrong, Jennifer Kim, Andrew Walkden, Leon Au, Arun Brahma, Fiona Carley, Jaya Devi Chidambaram
Scientific Reports.2025;[Epub] CrossRef
Assessing Acanthamoeba cytotoxicity: comparison of common cell viability assays
Alvie Loufouma Mbouaka, Iwona Lesiak-Markowicz, Irene Heredero-Bermejo, Rounik Mazumdar, Julia Walochnik, Tania Martín-Pérez
Frontiers in Microbiology.2023;[Epub] CrossRef
Host cell-type and pathogen-specific immunomodulatory functions of macrophage migration inhibitory factor (MIF) in infectious keratitis
Swagata Ghosh, AH Humera Khathun, G.S. Athulya, P. Vignesh, L Mathan, Ninad Mudaraddi, Siddharth Narendran, Prajna Lalitha, N. Venkatesh Prajna
Experimental Eye Research.2023; 236: 109669. CrossRef
Aspergillus fumigatus-Stimulated Human Corneal Epithelial Cells Induce Pyroptosis of THP-1 Macrophages by Secreting TSLP
Qingshan Ji, Lisong Wang, Jiajia Liu, Yali Wu, Huayi Lv, Yuechun Wen, Lei Shi, Bin Qu, Nóra Szentmáry
Inflammation.2021; 44(2): 682. CrossRef
Corneal Changes in Acanthamoeba Keratitis at Various Levels of Severity: An In Vivo Confocal Microscopic Study
Zhenyu Wei, Kai Cao, Leying Wang, Christophe Baudouin, Antoine Labbé, Qingfeng Liang
Translational Vision Science & Technology.2021; 10(7): 10. CrossRef
Polymicrobial Keratitis: Risk Factors, Clinical Characteristics, Bacterial Profile, and Antimicrobial Resistance
Laura A. González-Dibildox, José A. Oyervidez-Alvarado, Kristian A. Vazquez-Romo, Nallely Ramos-Betancourt, Everardo Hernandez-Quintela, Francisco Beltran, Manuel Garza-Leon
Eye & Contact Lens: Science & Clinical Practice.2021; 47(8): 465. CrossRef

8,066 View
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8 Web of Science
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Lethal Effects of Helianthemum lippii (L.) on Acanthamoeba castellanii Cysts in Vitro: F.A. Badria, M.H. Hetta, Rania M. Sarhan, M.H. Ezz El-Din; Korean J Parasitol 2014;52(3):243-249.
Published online June 26, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.3.243

Abstract
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Acanthamoeba spp. commonly cause Acanthamoeba keratitis which is typically associated with the wear of contact lenses. Therefore, finding an economic, efficient, and safe therapy of natural origin is of outmost importance. This study examined the in vitro lethal potential of ethyl acetate and methanol extracts of Helianthemum lippii (L.) (sun roses) against Acanthamoeba castellanii cysts isolated from patients with amoebic keratitis. Both extracts proved to be potent as regard to their lethal effects on A. castellanii cysts with comparable results to chlorhexidine. The ethyl acetate was more promising with cumulative lethality. It showed a highly significant lethal percentage along the duration of treatment. The analysis of the more potent ethyl acetate extract revealed the presence of 2.96 mg/100 g of total phenolics, 0.289 mg/100 ml of total flavonoids and 37 mg/100 mg of total tannins which highlighted their phytomedicinal role.

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Effectiveness of phytoproducts against pathogenic free-living amoebae - A scoping and critical review paving the way toward plant-based pharmaceuticals
Beni Jequicene Mussengue Chaúque, Thaisla Cristiane Borella da Silva, Eduardo Brittes Rott, Felipe Brittes Rott, Ana Paula Marçal Copetti Leite, Guilherme Brittes Benitez, Neuana Fernando Neuana, José Roberto Goldim, Marilise Brittes Rott, Régis Adriel Za
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Meyada Khaled, Rachid Ouache, Patrick Pale, Hassina Harkat
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Phytochemical on-line screening and in silico study of Helianthemum confertum: antioxidant activity, DFT, MD simulation, ADME/T analysis, and xanthine oxidase binding
Yasmine Chemam, Samir Benayache, Abdeslem Bouzina, Eric Marchioni, Omar Sekiou, Houria Bentoumi, Minjie Zhao, Zihad Bouslama, Nour-Eddine Aouf, Fadila Benayache
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Phytochemical-rich extracts of Helianthemum lippii possess antimicrobial, anticancer, and anti-biofilm activities
Fabiana Plescia, Fabio Venturella, Antonella D’Anneo, Valentina Catania, Maria Letizia Gargano, Giulia Polito, Domenico Schillaci, Antonio Palumbo Piccionello, Marianna Lauricella, Giuseppe Venturella, Demetrio Raffa
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Yassmin Isse Wehelie, Aishath Leesha Nasih, Ayaz Anwar, Ruqaiyyah Siddiqui, Sutherland Maciver, Naveed Ahmed Khan
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Maressa Dietrich Rosa, Jean Paulo de Andrade, Adriana Oliveira Costa, Raphael Conti, Jaume Bastida, Warley de Souza Borges, Cinthia Furst
Brazilian Journal of Pharmaceutical Sciences.2022;[Epub] CrossRef
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Ghada Mohamed Kadry, Mousa A. M. Ismail, Nagwa Mostafa El-Sayed, Hanan S. El-Kholy, Dina M. Hamdy El-Akkad
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Nihed Djemam, Somia Lassed, Fatih Gül, Muhammed Altun, Marisa Monteiro, Daniela Menezes-Pinto, Samir Benayache, Fadila Benayache, Djamila Zama, Ibrahim Demirtas, Manuela Morato
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Amoebicidal and Amoebistatic Effects of Artemisia argyi Methanolic Extracts on Acanthamoeba castellanii Trophozoites and Cysts
Onur Kolören, Zeynep Kolören, Zülal Atli Şekeroğlu, Melek Çolayvaz, Panagiotis Karanis
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Genetic Characterization of Clinical Acanthamoeba Isolates from Japan using Nuclear and Mitochondrial Small Subunit Ribosomal RNA: Md Moshiur Rahman, Kenji Yagita, Akira Kobayashi, Yosaburo Oikawa, Amjad I.A. Hussein, Takahiro Matsumura, Masaharu Tokoro; Korean J Parasitol 2013;51(4):401-411.
Published online August 30, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.4.401

Abstract
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Because of an increased number of Acanthamoeba keratitis (AK) along with associated disease burdens, medical professionals have become more aware of this pathogen in recent years. In this study, by analyzing both the nuclear 18S small subunit ribosomal RNA (18S rRNA) and mitochondrial 16S rRNA gene loci, 27 clinical Acanthamoeba strains that caused AK in Japan were classified into 3 genotypes, T3 (3 strains), T4 (23 strains), and T5 (one strain). Most haplotypes were identical to the reference haplotypes reported from all over the world, and thus no specificity of the haplotype distribution in Japan was found. The T4 sub-genotype analysis using the 16S rRNA gene locus also revealed a clear sub-conformation within the T4 cluster, and lead to the recognition of a new sub-genotype T4i, in addition to the previously reported sub-genotypes T4a-T4h. Furthermore, 9 out of 23 strains in the T4 genotype were identified to a specific haplotype (AF479533), which seems to be a causal haplotype of AK. While heterozygous nuclear haplotypes were observed from 2 strains, the mitochondrial haplotypes were homozygous as T4 genotype in the both strains, and suggested a possibility of nuclear hybridization (mating reproduction) between different strains in Acanthamoeba. The nuclear 18S rRNA gene and mitochondrial 16S rRNA gene loci of Acanthamoeba spp. possess different unique characteristics usable for the genotyping analyses, and those specific features could contribute to the establishment of molecular taxonomy for the species complex of Acanthamoeba.

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Molecular Evidence for Greater Diversity Within Acanthamoeba
Daniele Corsaro
Acta Parasitologica.2025;[Epub] CrossRef
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Kirti Megha, Megha Sharma, Amit Gupta, Rakesh Sehgal, Sumeeta Khurana
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The Risk Factors and Clinical Features of Acanthamoeba Keratitis: First Time Detection of Acanthamoeba T5 Genotype from Keratitis Patients in Turkey
Derya Dirim Erdogan, Mehmet Aykur, Nur Selvi Gunel, Melis Palamar, Ozlem Barut Selver, Buket Ozel, Ayse Yagci, Cumhur Gunduz, Hande Dagci
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Genotype distribution of Acanthamoeba in keratitis: a systematic review
Maria Luisa Nunes Diehl, Júlia Paes, Marilise Brittes Rott
Parasitology Research.2021; 120(9): 3051. CrossRef
Molecular identification and phylogenetic analysis of free-living amoeba (Naegleria and Acanthamoeba) from treated and untreated drinking water
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Keratitis by Acanthamoeba triangularis: Report of Cases and Characterization of Isolates: Ying-Hua Xuan, Byung-Suk Chung, Yeon-Chul Hong, Hyun-Hee Kong, Tae-Won Hahn, Dong-Il Chung; Korean J Parasitol 2008;46(3):157-164.
Published online September 20, 2008; DOI: https://doi.org/10.3347/kjp.2008.46.3.157

Abstract
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Three Acanthamoeba isolates (KA/E9, KA/E17, and KA/E23) from patients with keratitis were identified as Acanthamoeba triangularis by analysis of their molecular characteristics, a species not previously recognized to be a corneal pathogen. Epidemiologic significance of A. triangularis as a keratopathogen in Korea has been discussed. Morphologic features of Acanthamoeba cysts were examined under a microscope with differential interference contrast (DIC) optics. Mitochondrial DNA (mtDNA) of the ocular isolates KA/E9, KA/E17, and KA/E23 were digested with restriction enzymes, and the restriction patterns were compared with those of reference strains. Complete nuclear 18S and mitochondrial (mt) 16S rDNA sequences were subjected to phylogenetic analysis and species identification. mtDNA RFLP of 3 isolates showed very similar patterns to those of SH621, the type strain of A. triangularis. 16S and 18S rDNA sequence analysis confirmed 3 isolates to be A. triangularis. 18S rDNA sequence differences of the isolates were 1.3% to 1.6% and those of 16S rDNA, 0.4% to 0.9% from A. triangularis SH621. To the best of our knowledge, this is the first report, confirmed by 18S and 16S rDNA sequence analysis, of keratitis caused by A. triangularis of which the type strain was isolated from human feces. Six isolates of A. triangularis had been reported from contaminated contact lens cases in south eastern Korea.

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Molecular characterization of bacterial endosymbionts of Acanthamoeba isolates from infected corneas of Korean patients: Ying-Hua Xuan, Hak Sun Yu, Hae Jin Jeong, Sung-Yong Seol, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2007;45(1):1-9.
Published online March 20, 2007; DOI: https://doi.org/10.3347/kjp.2007.45.1.1

Abstract
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The endosymbionts of 4 strains of Acanthamoeba (KA/E9, KA/E21, KA/E22, and KA/E23) isolated from the infected corneas of Korean patients were characterized via orcein stain, transmission electron microscopic examination, and 16S rDNA sequence analysis. Double membrane-bound, rod-shaped endosymbionts were distributed randomly throughout both the trophozoites and cysts of each of Acanthamoeba isolates. The endosymbionts of KA/E9, KA/E22, and KA/E23 were surrounded by electron-translucent areas. No lacunae-like structures were observed in the endosymbionts of KA/E21, the bacterial cell walls of which were studded with host ribosomes. Comparative analyses of the 16S rDNA sequences showed that the endosymbionts of KA/E9, KA/E22 and KA/E23 were closely related to Caedibacter caryophilus, whereas the KA/E21 endosymbiont was assigned to the Cytophaga-Flavobacterium-Bacteroides (CFB) phylum. In the 4 strains of Acanthamoeba, the hosts of the endosymbionts were identified as belonging to the Acanthamoeba castellanii complex, which corresponds to the T4 genotype. Acanthamoeba KA/E21 evidenced characteristics almost identical to those of KA/E6, with the exception of the existence of endosymbionts. The discovery of these endosymbionts from Acanthamoeba may prove essential to future studies focusing on interactions between the endosymbionts and the amoebic hosts.

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Proteases of Acanthamoeba
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Comparison of specific activity and cytopathic effects of purified 33 kDa serine proteinase from Acanthamoeba strains with different degree of virulence: Won-Tae Kim, Hyun-Hee Kong, Young-Ran Ha, Yeon-Chul Hong, Hae Jin Jeong, Hak Sun Yu, Dong-Il Chung; Korean J Parasitol 2006;44(4):321-330.
Published online December 20, 2006; DOI: https://doi.org/10.3347/kjp.2006.44.4.321

Abstract
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The pathogenic mechanism of granulomatous amebic encephalitis (GAE) and amebic keratitis (AK) by Acanthamoeba has yet to be clarified. Protease has been recognized to play an important role in the pathogenesis of GAE and AK. In the present study, we have compared specific activity and cytopathic effects (CPE) of purified 33 kDa serine proteinases from Acanthamoeba strains with different degree of virulence (A. healyi OC-3A, A. lugdunensis KA/E2, and A. castellanii Neff). Trophozoites of the 3 strains revealed different degrees of CPE on human corneal epithelial (HCE) cells. The effect was remarkably reduced by adding phenylmethylsulfonylfluoride (PMSF), a serine proteinase inhibitor. This result indicated that PMSF-susceptible proteinase is the main component causing cytopathy to HCE cells by Acanthamoeba. The purified 33 kDa serine proteinase showed strong activity toward HCE cells and extracellular matrix proteins. The purified proteinase from OC-3A, the most virulent strain, demonstrated the highest enzyme activity compared to KA/E2, an ocular isolate, and Neff, a soil isolate. Polyclonal antibodies against the purified 33 kDa serine proteinase inhibit almost completely the proteolytic activity of culture supernatant of Acanthamoeba. In line with these results, the 33 kDa serine proteinase is suggested to play an important role in pathogenesis and to be the main component of virulence factor of Acanthamoeba.

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Molecular characterization of Acanthamoeba isolated from amebic keratitis related to orthokeratology lens overnight wear: Sun Joo Lee, Hae Jin Jeong, Ji Eun Lee, Jong Soo Lee, Ying Hua Xuan, Hyun-Hee Kong, Dong-Il Chung, Mee-Sun Ock, Hak Sun Yu; Korean J Parasitol 2006;44(4):313-320.
Published online December 20, 2006; DOI: https://doi.org/10.3347/kjp.2006.44.4.313

Abstract
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In an effort to characterize, on the molecular scale, the Acanthamoeba initially isolated from the cornea of an amoebic keratitis patient associated with overnight-wear orthokeratology lens in Korea, we conducted mitochondrial DNA restriction fragment length polymorphism, 18S rDNA sequencing, and drug sensitivity analyses on the isolate (KA/PE1). The patient was treated with polyhexamethylene biguanide, chlorhexidine and oral itraconazole, which resulted in resolution of the patient's ocular inflammation. The majority of the molecular characteristics of the KA/PE1 were determined to be identical, or quite similar, to those of A. castellanii Ma strain, which had been isolated also from amoebic keratitis. The risk of Acanthamoeba keratitis as a potential complication of overnight orthokeratology is briefly discussed.

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Brief Communication

In vitro cytotoxicity of Acanthamoeba spp. isolated from contact lens containers in Korea by crystal violet staining and LDH release assay: Ho-Joon Shin, Myung-Soo Cho, Suk-Yul Jung, Hyung-Il Kim, Kyung-il Im; Korean J Parasitol 2000;38(2):99-102.
Published online June 30, 2000; DOI: https://doi.org/10.3347/kjp.2000.38.2.99

Abstract
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In order to observe the cytotoxicity of Acanthamoeba spp., which were isolated from contact lens containers as ethiological agents for the probable amoebic keratitis in Korea, the crystal violet staining method and LDH release assay were carried out. In the crystal violet staining method, among eight contact lens container isolates, isolate 3 (Acanthamoeba KA/LS5) showed 83.6% and 81.8% of cytotoxicity, and isolate 7 (Acanthamoeba KA/LS37) showed 28.2% and 25.1% of cytotoxicity, in 1 mg/ml and 0.5 mg/ml lysate treatments, respectively. Acanthamoeba culbertsoni and A. healyi showed 84.0% and 82.8% of cytotoxicity. Similar results were observed in A. castellanii and A. hatchetti which showed 83.6% and 75.5% of cytotoxicity. Acanthamoeba royreba and A. polyphaga showed 9.0% and 1.7% of cytotoxicity. In the LDH release assay, isolate 3 (20.4%) showed higher cytotoxicity than other isolates in 1 mg/ml lysate treatment. The results provide that at least isolate 3 has the cytotoxic effect against CHO cells and seems to be the pathogenic strain.

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Original Article

Isoenzyme patterns and phylogenetic relationships in Acanthamoeba spp. isolated from contact lens containers in Korea: Ho-Joon Shin, Myung-Soo Cho, Han-Jip Kim, Kyung-Il Im; Korean J Parasitol 1999;37(4):229-236.
Published online December 31, 1999; DOI: https://doi.org/10.3347/kjp.1999.37.4.229

Abstract
PDF

In order to refer to the basic information regarding the identification of isolates obtained from a contact lens container in Korea, the isoelectric focusing gel electrophoresis was employed to compare the isoenzyme band patterns among Acanthamoeba spp. including eight isolates and the simple pairwise dissimilarity analysis was carried out. For an alkaline phosphate development, isolate 7 and Acanthamoeba polyphaga showed homologous band patterns, and isolates 1, 2, and 3 showed the same patterns. For lactate dehydrogenase, similar patterns were observed in isolates 2 and 3. Isolates 3 and 5 showed homologous band patterns for malate dehydrogenase and glucose phosphate isomerase. For hexokinase, isolates 4, 7, and A. hatchetti showed the same band patterns. In others, a considerable number of interstrain polymorphisms was observed in nine isoenzyme band patterns. In Acanthamoeba group II, genetic distances among isolates 1, 2, 3, 4, and 5 ranged from 0.104 to 0.200. In comparison to A. castellanii, A. hatchetti, and A. polyphaga, genetic distances of isolates 7 and 8 were 0.254 and 0.219, respectively. In Acanthamoeba group III, including A. culbertsoni, A. healyi, and A. royreba, isolate 6 had genetic distances which ranged from 0.314 to 0.336. Finally, when comparing to the six reference Acanthamoeba, it was possible to classify isolates 1, 2, 3, 4, and 5, as genetically close-related species and as independent species group. Furthermore, isolates 6, 7 and 8 were identified as independent species as well.

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