Parasites, Hosts and Diseases

"Hyun-Hee Kong"

Original Articles

Identification of essential genes for Acanthamoeba castellanii excystation during encystation and excystation: Min-Jeong Kim, Hye-Jeong Jo, Fu-Shi Quan, Ki Back Chu, Hyun-Hee Kong, Eun-Kyung Moon; Parasites Hosts Dis 2024;62(4):399-407.
Published online November 22, 2024; DOI: https://doi.org/10.3347/PHD.24062

Acanthamoeba is an opportunistic pathogen that causes Acanthamoeba keratitis, granulomatous amoebic encephalitis, and other cutaneous diseases. The life cycle of Acanthamoeba consists of 2 stages of trophozoites and cysts. Under adverse environmental conditions, Acanthamoeba encysts, while the conditions become favorable for growth, it reverts to the trophozoite form. Acanthamoeba excystation is crucial for its proliferation and can lead to recurrent infections after incomplete treatment. To identify the factors involved in excystation, A. castellanii was subjected to either encystation- or excystation-inducing conditions, and gene expression profiles were compared using mRNA sequencing. A. castellanii samples were collected at 8 h intervals for analysis under both conditions. Differentially expressed gene analysis revealed that 1,214 and 1,163 genes were upregulated and downregulated, respectively, by more than 2-fold during early excystation. Five genes markedly upregulated in early excystation (ACA1_031140, ACA1_032330, ACA1_374400, ACA1_275740, and ACA1_112650) were selected, and their expression levels were confirmed via real-time PCR. Small interfering RNA (siRNA) targeting these 5 genes was transfected into Acanthamoeba and gene knockdown was validated through real-time PCR. The silencing of ACA1_031140, ACA1_032330, ACA1_374400, and ACA1_112650 inhibited excystation and suggested that these genes might be essential for excystation. Our findings provide valuable insights for suppressing Acanthamoeba proliferation and recurrence.

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Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
Mathew Choaji, Ascel Samba-Louaka, Zineb Fechtali-Moute, Willy Aucher, Sébastien Pomel
Pathogens.2025; 14(3): 268. CrossRef
Integrating molecular pathogenesis and predictive modelling in granulomatous amoebic encephalitis due to Acanthamoeba : a digital twin framework for personalised therapy
Ruqaiyyah Siddiqui, David Lloyd, Naveed Ahmed Khan
Infectious Diseases.2025; : 1. CrossRef
Microarray-based characterization of airway inflammation induced by environmental Acanthamoeba exposure versus the ovalbumin-alum model
Shin Ae Kang, Hak Sun Yu
Parasites, Hosts and Diseases.2025; 63(4): 327. CrossRef

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Phagocytosis-associated genes in Acanthamoeba castellanii feeding on Escherichia coli: Min-Jeong Kim, Eun-Kyung Moon, Hye-Jeong Jo, Fu-Shi Quan, Hyun-Hee Kong; Parasites Hosts Dis 2023;61(4):397-404.
Published online November 28, 2023; DOI: https://doi.org/10.3347/PHD.23088

Acanthamoeba species are free-living amoebae those are widely distributed in the environment. They feed on various microorganisms, including bacteria, fungi, and algae. Although majority of the microbes phagocytosed by Acanthamoeba spp. are digested, some pathogenic bacteria thrive within them. Here, we identified the roles of 3 phagocytosis-associated genes (ACA1_077100, ACA1_175060, and AFD36229.1) in A. castellanii. These 3 genes were upregulated after the ingestion of Escherichia coli. However, after the ingestion of Legionella pneumophila, the expression of these 3 genes was not altered after the consumption of L. pneumophila. Furthermore, A. castellanii transfected with small interfering RNS (siRNA) targeting the 3 phagocytosis-associated genes failed to digest phagocytized E. coli. Silencing of ACA1_077100 disabled phagosome formation in the E. coli-ingesting A. castellanii. Alternatively, silencing of ACA1_175060 enabled phagosome formation; however, phagolysosome formation was inhibited. Moreover, suppression of AFD36229.1 expression prevented E. coli digestion and consequently led to the rupturing of A. castellanii. Our results demonstrated that the ACA1_077100, ACA1_175060, and AFD36229.1 genes of Acanthamoeba played crucial roles not only in the formation of phagosome and phagolysosome but also in the digestion of E. coli.

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Phylogenomic, structural, and cell biological analyses reveal that Stenotrophomonas maltophilia replicates in acidified Rab7A-positive vacuoles of Acanthamoeba castellanii
Javier Rivera, Julio C. Valerdi-Negreros, Diana M. Vázquez-Enciso, Fulvia-Stefany Argueta-Zepeda, Pablo Vinuesa, Michael L. Ginger, Monica Crary, Sutherland K. Maciver
Microbiology Spectrum.2024;[Epub] CrossRef

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Trichinella Infection Ameliorated Vincristine-Induced Neuroinflammation in Mice: Young Rae Jo, Hwan Tae Park, Hak Sun Yu, Hyun-Hee Kong; Korean J Parasitol 2022;60(4):247-254.
Published online August 24, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.4.247

Abstract
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Vincristine (VCR) is a chemotherapeutic agent widely used in treatment of malignancies. However, VCR has a limitation in use since it commonly causes a painful neuropathy (VCR-induced peripheral neuropathy, VIPN). Inflammatory cytokines secreted by immune cells such as macrophages can exacerbate allodynia and hyperalgesia, because inhibiting the inflammatory response is a treatment target for VIPN. In this study, we investigated whether Trichinella spiralis, a widely studied helminth for its immunomodulatory abilities, can alleviate VCR-induced allodynia. Von Frey test showed that T. spiralis infection improved mechanical allodynia at 10 days after VCR injection. We further observed whether the difference was due to mitigated axon degeneration, but no significant difference between the groups in axonal degeneration in sciatic nerves and intra-epidermal nerve fibers was found. Conversely, we observed that number of infiltrated macrophages was decreased in the sciatic nerves of the T. spiralis infected mice. Moreover, treatment of T. spiralis excretory-secretory products caused peritoneal macrophages to secrete decreased level of IL-1β. This study suggests that T. spiralis can relieve VCR-induced mechanical allodynia by suppressing neuroinflammation and that application of controllable degree of helminth may prove beneficial for VIPN treatment.

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Therapeutic potentials of Trichinella spiralis in immune disorders: From allergy to autoimmunity
Minkyoung Cho, Hak Sun Yu
Parasites, Hosts and Diseases.2025; 63(2): 123. CrossRef
Alleviation of Rheumatoid Arthritis by Inducing IDO Expression with Trichinella spiralis Recombinant Protein 43
Xiao Ma, Dongming Liu, Wenhao Yu, Caixia Han, Arif Siddiqui
Journal of Immunology Research.2024;[Epub] CrossRef
Anti-Inflammatory Effects of Helminth-Derived Products: Potential Applications and Challenges in Diabetes Mellitus Management
Yunhuan Zhu, Xintong Chen, Hezheng Zheng, Qiman Ma, Keda Chen, Hongyu Li
Journal of Inflammation Research.2024; Volume 17: 11789. 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.

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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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Original Articles

Characterization of a Peptide Antibody Specific to the Adenylyl Cyclase-Associated Protein of Acanthamoeba castellanii: Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon; Korean J Parasitol 2022;60(1):7-14.
Published online February 23, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.1.7

Abstract
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Acanthamoeba keratitis (AK) is a rare infectious disease and accurate diagnosis has remained arduous as clinical manifestations of AK were similar to keratitis of viral, bacterial, or fungal origins. In this study, we described the production of a polyclonal peptide antibody against the adenylyl cyclase-associated protein (ACAP) of A. castellanii, and evaluated its differential diagnostic potential. Enzyme-linked immunosorbent assay revealed high titers of A. castellanii-specific IgG and IgA antibodies being present in low dilutions of immunized rabbit serum. Western blot analysis revealed that the ACAP antibody specifically interacted with A. castellanii, while not interacting with human corneal epithelial (HCE) cells and other causes of keratitis such as Fusarium solani, Pseudomonas aeruginosa, and Staphylococcus aureus. Immunocytochemistry (ICC) results confirmed the specific detection of trophozoites and cysts of A. castellanii co-cultured with HCE cells. The ACAP antibody also specifically interacted with the trophozoites and cysts of 5 other Acanthamoeba species. These results indicate that the ACAP antibody of A. castellanii can specifically detect multiple AK-causing members belonging to the genus Acanthamoeba and may be useful for differentially diagnosing Acanthamoeba infections.

Citations

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The pathogenesis, risk factors, diagnosis and treatment of Acanthamoeba keratitis
Mingliang Bao, Hai Bao, Shuqing Wang, Hongyan Zhou
Frontiers in Medicine.2025;[Epub] 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
New Frontiers in Acanthamoeba Keratitis Diagnosis and Management
Omar Shareef, Sana Shareef, Hajirah N. Saeed
Biology.2023; 12(12): 1489. 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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Sirtinol Supresses Trophozoites Proliferation and Encystation of Acanthamoeba via Inhibition of Sirtuin Family Protein: So-Young Joo, Ja Moon Aung, Minsang Shin, Eun-Kyung Moon, Hyun-Hee Kong, Youn-Kyoung Goo, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2022;60(1):1-6.
Published online February 23, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.1.1

Abstract
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The encystation of Acanthamoeba leads to the development of metabolically inactive and dormant cysts from vegetative trophozoites under unfavorable conditions. These cysts are highly resistant to anti-Acanthamoeba drugs and biocides. Therefore, the inhibition of encystation would be more effective in treating Acanthamoeba infection. In our previous study, a sirtuin family protein—Acanthamoeba silent-information regulator 2-like protein (AcSir2)—was identified, and its expression was discovered to be critical for Acanthamoeba castellanii proliferation and encystation. In this study, to develop Acanthamoeba sirtuin inhibitors, we examine the effects of sirtinol, a sirtuin inhibitor, on trophozoite growth and encystation. Sirtinol inhibited A. castellanii trophozoites proliferation (IC50=61.24 μM). The encystation rate of cells treated with sirtinol significantly decreased to 39.8% (200 μM sirtinol) after 24 hr of incubation compared to controls. In AcSir2-overexpressing cells, the transcriptional level of cyst-specific cysteine protease (CSCP), an Acanthamoeba cysteine protease involved in the encysting process, was 11.6- and 88.6-fold higher at 48 and 72 hr after induction of encystation compared to control. However, sirtinol suppresses CSCP transcription, resulting that the undegraded organelles and large molecules remained in sirtinol-treated cells during encystation. These results indicated that sirtinol sufficiently inhibited trophozoite proliferation and encystation, and can be used to treat Acanthamoeba infections.

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Pterostilbene: A natural neuroprotective stilbene with anti-Alzheimer's disease properties
Songlan Gao, Honglei Zhang, Na Li, Lijuan Zhang, Zhe Zhu, Changlu Xu
Journal of Pharmaceutical Analysis.2025; 15(4): 101043. CrossRef
Alzheimer’s Disease: A Review of Molecular Mechanisms and Therapeutic Implications by Targeting Sirtuins, Caspases, and GSK-3
Kalpana Pandya, Krishnashish Roul, Avanish Tripathi, Sateesh Belemkar, Anshuman Sinha, Meryem Erol, Devendra Kumar
ACS Chemical Neuroscience.2025; 16(12): 2178. CrossRef
Human Conjunctival Transcriptome in Acanthamoeba Keratitis: An Exploratory Study
Gerami D. Seitzman, Jeremy D. Keenan, Thomas M. Lietman, Kevin Ruder, Lina Zhong, Cindi Chen, YuHeng Liu, Danny Yu, Thomas Abraham, Armin Hinterwirth, Thuy Doan
Cornea.2024; 43(10): 1272. CrossRef
Comparative cytotoxicity of Acanthamoeba castellanii-derived conditioned medium on human corneal epithelial and stromal cells
Abdullah Alhazmi, Laura E. Sidney, Andy Hopkinson, Hany M. Elsheikha
Acta Tropica.2024; 257: 107288. CrossRef
Biological characteristics and pathogenicity of Acanthamoeba
Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
Frontiers in Microbiology.2023;[Epub] CrossRef

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Brief Communication

Differentially Expressed Gene Profile of Acanthamoeba castellanii Induced by an Endosymbiont Legionella pneumophila: Eun-Kyung Moon, So-Min Park, Ki-Back Chu, Fu-Shi Quan, Hyun-Hee Kong; Korean J Parasitol 2021;59(1):67-75.
Published online February 19, 2021; DOI: https://doi.org/10.3347/kjp.2021.59.1.67

Abstract
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Legionella pneumophila is an opportunistic pathogen that survives and proliferates within protists such as Acanthamoeba spp. in environment. However, intracellular pathogenic endosymbiosis and its implications within Acanthamoeba spp. remain poorly understood. In this study, RNA sequencing analysis was used to investigate transcriptional changes in A. castellanii in response to L. pneumophila infection. Based on RNA sequencing data, we identified 1,211 upregulated genes and 1,131 downregulated genes in A. castellanii infected with L. pneumophila for 12 hr. After 24 hr, 1,321 upregulated genes and 1,379 downregulated genes were identified. Gene ontology (GO) analysis revealed that L. pneumophila endosymbiosis enhanced hydrolase activity, catalytic activity, and DNA binding while reducing oxidoreductase activity in the molecular function (MF) domain. In particular, multiple genes associated with the GO term ‘integral component of membrane’ were downregulated during endosymbiosis. The endosymbiont also induced differential expression of various methyltransferases and acetyltransferases in A. castellanii. Findings herein are may significantly contribute to understanding endosymbiosis of L. pneumophila within A. castellanii.

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Oxford Nanopore Technology-Based Identification of an Acanthamoeba castellanii Endosymbiosis in Microbial Keratitis
Sebastian Alexander Scharf, Lennart Friedrichs, Robert Bock, Maria Borrelli, Colin MacKenzie, Klaus Pfeffer, Birgit Henrich
Microorganisms.2024; 12(11): 2292. CrossRef
Transcription dynamics of heat-shock proteins (Hsps) and endosymbiont titres in response to thermal stress in whitefly, Bemisia tabaci (Asia-I)
Mritunjoy Barman, Snigdha Samanta, Bulbul Ahmed, Soumik Dey, Swati Chakraborty, M.G. Deeksha, Subham Dutta, Arunava Samanta, Jayanta Tarafdar, Deepayan Roy
Frontiers in Physiology.2023;[Epub] CrossRef
Proteomic analysis of Acanthamoeba castellanii response to Legionella pneumophila infection
Alban Hay, Steven Rolland, Clément Bernard, Yann Héchard, Romain Villéger, Ascel Samba-Louaka
FEMS Microbiology Letters.2023;[Epub] CrossRef
Comparative analysis of differentially expressed genes in Acanthamoeba after ingestion of Legionella pneumophila and Escherichia coli
Eun-Kyung Moon, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
Experimental Parasitology.2022; 232: 108188. CrossRef

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Original Article

Comparison of Proteins Secreted into Extracellular Space of Pathogenic and Non-pathogenic Acanthamoeba castellanii: Eun-Kyung Moon, Hyun-Seo Choi, So-Min Park, Hyun-Hee Kong, Fu-Shi Quan; Korean J Parasitol 2018;56(6):553-558.
Published online December 31, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.6.553

Pathogenic Acanthamoeba spp. cause granulomatous amoebic encephalitis and keratitis. Acanthamoeba keratitis (AK) is a rare but serious ocular infection that can result in permanent visual impairment or blindness. However, pathogenic factors of AK remain unclear and treatment for AK is arduous. Expression levels of proteins secreted into extracellular space were compared between A. castellanii pathogenic (ACP) and non-pathogenic strains. Two-dimensional polyacrylamide gel electrophoresis revealed 123 differentially expressed proteins, including 34 increased proteins , 7 qualitative increased proteins, 65 decreased proteins, and 17 qualitative decreased proteins in ACP strain. Twenty protein spots with greater than 5-fold increase in ACP strain were analyzed by liquid chromatography triple quadrupole mass spectrometry. These proteins showed similarity each to inosine-uridine preferring nucleoside hydrolase, carboxylesterase, oxygen-dependent choline dehydrogenase, periplasmic-binding protein proteinases and hypothetical proteins. These proteins expressed higher in ACP may provide some information to understand pathogenicity of Acanthamoeba.

Citations

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The gene expression and proteomic profiling of Acanthamoeba isolates
Chayan Sharma, Sumeeta Khurana, Alka Bhatia, Amit Arora, Amit Gupta
Experimental Parasitology.2023; 255: 108630. CrossRef
Detection of Acanthamoeba spp. using carboxylesterase antibody and its usage for diagnosing Acanthamoeba-keratitis
Min-Jeong Kim, Ki-Back Chu, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon, Ashok Kumar
PLOS ONE.2022; 17(1): e0262223. CrossRef
Characterization of a Peptide Antibody Specific to the Adenylyl Cyclase-Associated Protein of Acanthamoeba castellanii
Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong, Eun-Kyung Moon
The Korean Journal of Parasitology.2022; 60(1): 7. CrossRef
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
The Korean Journal of Parasitology.2022; 60(2): 143. CrossRef
Host-Parasite Interaction of Atlantic salmon (Salmo salar) and the Ectoparasite Neoparamoeba perurans in Amoebic Gill Disease
Natasha A. Botwright, Amin R. Mohamed, Joel Slinger, Paula C. Lima, James W. Wynne
Frontiers in Immunology.2021;[Epub] CrossRef
Proteomic analysis of the skeletal muscles from dysferlinopathy patients
Young-Chul Choi, Ji-Man Hong, Kee Duk Park, Ha Young Shin, Seung Min Kim, Hyung Jun Park
Journal of Clinical Neuroscience.2020; 71: 186. CrossRef
Production of a polyclonal antibody against inosine-uridine preferring nucleoside hydrolase of Acanthamoeba castellanii and its access to diagnosis of Acanthamoeba keratitis
So-Min Park, Hae-Ahm Lee, Ki-Back Chu, Fu-Shi Quan, Su-Jung Kim, Eun-Kyung Moon, Paulo Lee Ho
PLOS ONE.2020; 15(9): e0239867. CrossRef
Quantitative proteomic analysis and functional characterization of Acanthamoeba castellanii exosome-like vesicles
Wei-Chen Lin, Chia-Yun Tsai, Jian-Ming Huang, Shang-Rung Wu, Lichieh Julie Chu, Kuo-Yang Huang
Parasites & Vectors.2019;[Epub] CrossRef

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Brief Communication

Effect of 2, 6-Dichlorobenzonitrile on Amoebicidal Activity of Multipurpose Contact Lens Disinfecting Solutions: Eun-Kyung Moon, Seungeun Lee, Fu-Shi Quan, Hyun-Hee Kong; Korean J Parasitol 2018;56(5):491-494.
Published online October 31, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.5.491

Multipurpose contact lens disinfecting solutions (MPDS) are widely used to cleanse and disinfect microorganisms. However, disinfection efficacy of these MPDS against Acanthamoeba cyst remain insufficient. 2, 6-dichlorobenzonitrile (DCB), a cellulose synthesis inhibitor, is capable of increasing the amoebical effect against Acanthamoeba by inhibiting its encystation. In this study, we investigated the possibility of DCB as a disinfecting agent to improve the amoebicidal activity of MPDS against Acanthamoeba cyst. Eight commercial MPDS (from a to h) were assessed, all of which displayed insufficient amoebicidal activity against the mature cysts. Solution e, f, and h showed strong amoebicidal effect on the immature cysts. Amoebicidal efficacy against mature cysts remained inadequate even when the 8 MPDS were combined with 100 μM DCB. However, 4 kinds of MPDS (solution d, e, f, and h) including 100 μM DCB demonstrated strong amoebicidal activity against the immature cysts. The amoebicidal activity of solution d was increased by addition of DCB. Cytotoxicity was absent in human corneal epithelial cells treated with either DCB or mixture of DCB with MPDS. These results suggested that DCB can enhance the amoebicical activity of MPDS against Acanthamoeba immature cyst in vitro.

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Simultaneous quantitative determination of low-concentration ternary pesticide mixtures in wheat flour based on terahertz spectroscopy and BPNN
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Acanthamoeba keratitis: a review of biology, pathophysiology and epidemiology
Ana Gomes de Lacerda, Madalena Lira
Ophthalmic and Physiological Optics.2021; 41(1): 116. CrossRef
Differential Antimicrobial Efficacy of Multipurpose Solutions against Acanthamoeba Trophozoites
Rhonda Walters, Elise Miller, Allison Campolo, Manal M. Gabriel, Paul Shannon, Cindy McAnally, Monica Crary
Optometry and Vision Science.2021; 98(12): 1379. CrossRef
Drug Discovery against Acanthamoeba Infections: Present Knowledge and Unmet Needs
Hany M. Elsheikha, Ruqaiyyah Siddiqui, Naveed Ahmed Khan
Pathogens.2020; 9(5): 405. CrossRef

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Original Articles

DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation: Eun-Kyung Moon, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong; Korean J Parasitol 2017;55(2):115-120.
Published online April 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.2.115

Abstract
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Encystation mediating cyst specific cysteine proteinase (CSCP) of Acanthamoeba castellanii is expressed remarkably during encystation. However, the molecular mechanism involved in the regulation of CSCP gene expression remains unclear. In this study, we focused on epigenetic regulation of gene expression during encystation of Acanthamoeba. To evaluate methylation as a potential mechanism involved in the regulation of CSCP expression, we first investigated the correlation between promoter methylation status of CSCP gene and its expression. A 2,878 bp of promoter sequence of CSCP gene was amplified by PCR. Three CpG islands (island 1-3) were detected in this sequence using bioinformatics tools. Methylation of CpG island in trophozoites and cysts was measured by bisulfite sequence PCR. CSCP promoter methylation of CpG island 1 (1,633 bp) was found in 8.2% of trophozoites and 7.3% of cysts. Methylation of CpG island 2 (625 bp) was observed in 4.2% of trophozoites and 5.8% of cysts. Methylation of CpG island 3 (367 bp) in trophozoites and cysts was both 3.6%. These results suggest that DNA methylation system is present in CSCP gene expression of Acanthamoeba. In addition, the expression of encystation mediating CSCP is correlated with promoter CpG island 1 hypomethylation.

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DNA methylation modification: Dawn of research on cornea-related diseases
Quanhao Pan, Xiaoning Ge, Di Wang, Yuxi He
Life Sciences.2025; 376: 123757. CrossRef
Proteases of Acanthamoeba
Behroz Mahdavi Poor, Jalil Rashedi, Vahid Asgharzadeh, Amirali Mirmazhary, Nazila Gheitarani
Parasitology Research.2024;[Epub] CrossRef
Acanthamoeba keratitis: new hopes for potential interventions for a curable but often refractory disease
Bader Saleem Alawfi, Naveed Ahmed Khan, David Lloyd, Ruqaiyyah Siddiqui
Expert Review of Ophthalmology.2024; 19(4): 271. CrossRef
Biological characteristics and pathogenicity of Acanthamoeba
Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
Frontiers in Microbiology.2023;[Epub] CrossRef
Coevolution of the CDCA7-HELLS ICF-related nucleosome remodeling complex and DNA methyltransferases
Hironori Funabiki, Isabel E Wassing, Qingyuan Jia, Ji-Dung Luo, Thomas Carroll
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Hironori Funabiki, Isabel E Wassing, Qingyuan Jia, Ji-Dung Luo, Thomas Carroll
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Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii: Eun-Kyung Moon, Hyun-Hee Kong, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan; Korean J Parasitol 2017;55(2):109-114.
Published online April 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.2.109

Abstract
PDF

Protein arginine methyltransferase (PRMT) is an important epigenetic regulator in eukaryotic cells. During encystation, an essential process for Acanthamoeba survival, the expression of a lot of genes involved in the encystation process has to be regulated in order to be induced or inhibited. However, the regulation mechanism of these genes is yet unknown. In this study, the full-length 1,059 bp cDNA sequence of Acanthamoeba castellanii PRMT1 (AcPRMT1) was cloned for the first time. The AcPRMT1 protein comprised of 352 amino acids with a SAM-dependent methyltransferase PRMT-type domain. The expression level of AcPRMT1 was highly increased during encystation of A. castellanii. The EGFPAcPRMT1 fusion protein was distributed over the cytoplasm, but it was mainly localized in the nucleus of Acanthamoeba. Knock down of AcPRMT1 by synthetic siRNA with a complementary sequence failed to form mature cysts. These findings suggested that AcPRMT1 plays a critical role in the regulation of encystation of A. castellanii. The target gene of AcPRMT1 regulation and the detailed mechanisms need to be investigated by further studies.

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Drug Discovery against Acanthamoeba Infections: Present Knowledge and Unmet Needs
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Free-living amoebae and squatters in the wild: ecological and molecular features
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FEMS Microbiology Reviews.2019; 43(4): 415. CrossRef

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Efficacy of Korean Multipurpose Contact Lens Disinfecting Solutions against Acanthamoeba castellanii: Eun-Kyung Moon, Hye-Ryun Park, Fu-Shi Quan, Hyun-Hee Kong; Korean J Parasitol 2016;54(6):697-702.
Published online December 31, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.6.697

Abstract
PDF

Acanthamoeba keratitis has been increasing in recent years. Main risk factors are contact lens wear and their cleaning solutions. Most contact lens wearers use multipurpose disinfecting solutions (MPDS) for cleansing and disinfecting microorganisms because of its convenience. We determined amoebicidal effects of MPDS made in Korea and their cytotoxicity on human corneal epithelium cells. Fifteen commercial MPDS (A to O) were tested for their amoebicidal effects on Acanthamoeba castellanii trophozoites and cysts by using a most probable number (MPN) technique. Among them, 7 kinds of MPDS showed little or no amoebicidal effects for 24 hr exposure. Solutions A, B, G, H, L, and O showed positive amoebicidal effects, and solutions M and N killed almost all trophozoites and cysts after 24 hr exposure. However, 50%-N solution showed 56% cytotoxicity on human corneal epithelial cells within 4 hr exposure, and 50%-O solution also showed 62% cytotoxicity on human cells within 4 hr exposure. Solution A did not show any cytotoxicity on human cells. These results revealed that most MPDS made in Korea were ineffective to kill Acanthamoeba. The solutions having amoebicidal activity also showed high levels of cytotoxicity on human corneal epithelial cells. New formulations for improved MPDS that are amoebicidal but safe for host cells are needed to prevent Acanthamoeba keratitis.

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Seung-Mok Lee, Da-In Lee, Sung-Hee Park, Hak-Sun Yu, Ji-Eun Lee, Jong-Soo Lee
Cornea.2017; 36(12): 1538. CrossRef

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Brief Communication

Loop-Mediated Isothermal Amplification Targeting Actin DNA of Trichomonas vaginalis: Youn-Kyoung Goo, Won-Sik Shin, Hye-Won Yang, So-Young Joo, Su-Min Song, Jae-Sook Ryu, Hyun-Hee Kong, Won-Ki Lee, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2016;54(3):329-334.
Published online June 30, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.3.329

Abstract
PDF

Trichomoniasis caused by Trichomonas vaginalis is a common sexually transmitted disease. Its association with several health problems, including preterm birth, pelvic inflammatory disease, cervical cancer, and transmission of human immunodeficiency virus, emphasizes the importance of improved access to early and accurate detection of T. vaginalis. In this study, a rapid and efficient loop-mediated isothermal amplification-based method for the detection of T. vaginalis was developed and validated, using vaginal swab specimens from subjects suspected to have trichomoniasis. The LAMP assay targeting the actin gene was highly sensitive with detection limits of 1 trichomonad and 1 pg of T. vaginalis DNA per reaction, and specifically amplified the target gene only from T. vaginalis. Validation of this assay showed that it had the highest sensitivity and better agreement with PCR (used as the gold standard) compared to microscopy and multiplex PCR. This study showed that the LAMP assay, targeting the actin gene, could be used to diagnose early infections of T. vaginalis. Thus, we have provided an alternative molecular diagnostic tool and a point-of-care test that may help to prevent trichomoniasis transmission and associated complications.

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Construction a novel detection method for Trichomonas vaginalis based on recombinant enzyme polymerase amplification targeting the Actin gene
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Label-free ultrasensitive electrochemical genosensing of Trichomonas vaginalis using anisotropic-shaped gold nanoparticles as a platform, a repeated sequence of the parasite DNA as a probe, and toluidine blue as a redox marker
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Original Article

Identification of Protein Arginine Methyltransferase 5 as a Regulator for Encystation of Acanthamoeba: Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong; Korean J Parasitol 2016;54(2):133-138.
Published online April 30, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.2.133

Abstract
PDF

Encystation is an essential process for Acanthamoeba survival under nutrient-limiting conditions and exposure to drugs. The expression of several genes has been observed to increase or decrease during encystation. Epigenetic processes involved in regulation of gene expression have been shown to play a role in several pathogenic parasites. In the present study, we identified the protein arginine methyltransferase 5 (PRMT5), a known epigenetic regulator, in Acanthamoeba castellanii. PRMT5 of A. castellanii (AcPRMT5) contained domains found in S-adenosylmethionine-dependent methyltransferases and in PRMT5 arginine-N-methyltransferase. Expression levels of AcPRMT5 were increased during encystation of A. castellanii. The EGFP-PRMT5 fusion protein was mainly localized in the nucleus of trophozoites. A. castellanii transfected with siRNA designed against AcPRMT5 failed to form mature cysts. The findings of this study lead to a better understanding of epigenetic mechanisms behind the regulation of encystation in cyst-forming pathogenic protozoa.

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Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
Mathew Choaji, Ascel Samba-Louaka, Zineb Fechtali-Moute, Willy Aucher, Sébastien Pomel
Pathogens.2025; 14(3): 268. CrossRef
Oxidase enzyme genes are differentially expressed during Acanthamoeba castellanii encystment
Christian Q. Scheckhuber, Rebeca Damián Ferrara, Jesús Gómez-Montalvo, Sutherland K. Maciver, Alvaro de Obeso Fernández del Valle
Parasitology Research.2024;[Epub] CrossRef
Acanthamoeba keratitis: new hopes for potential interventions for a curable but often refractory disease
Bader Saleem Alawfi, Naveed Ahmed Khan, David Lloyd, Ruqaiyyah Siddiqui
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Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
Frontiers in Microbiology.2023;[Epub] CrossRef
Comparative analysis of differentially expressed genes in Acanthamoeba after ingestion of Legionella pneumophila and Escherichia coli
Eun-Kyung Moon, Min-Jeong Kim, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
Experimental Parasitology.2022; 232: 108188. CrossRef
Stimulation of Acanthamoeba castellanii excystment by enzyme treatment and consequences on trophozoite growth
Zineb Fechtali-Moute, Philippe M. Loiseau, Sébastien Pomel
Frontiers in Cell and Developmental Biology.2022;[Epub] CrossRef
Free-living amoebae and squatters in the wild: ecological and molecular features
Ascel Samba-Louaka, Vincent Delafont, Marie-Hélène Rodier, Estelle Cateau, Yann Héchard
FEMS Microbiology Reviews.2019; 43(4): 415. CrossRef
Encystation: the most prevalent and underinvestigated differentiation pathway of eukaryotes
Pauline Schaap, Christina Schilde
Microbiology.2018; 164(5): 727. CrossRef
Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii
Eun-Kyung Moon, Hyun-Hee Kong, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan
The Korean Journal of Parasitology.2017; 55(2): 109. CrossRef
DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation
Eun-Kyung Moon, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
The Korean Journal of Parasitology.2017; 55(2): 115. CrossRef

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Brief Communications

Prevalence of Trichomonas vaginalis in Women Visiting 2 Obstetrics and Gynecology Clinics in Daegu, South Korea: Youn-Kyoung Goo, Won-Sik Shin, Hye-Won Yang, So-Young Joo, Su-Min Song, Jae-Sook Ryu, Won-Myung Lee, Hyun-Hee Kong, Won-Ki Lee, Sang-Eun Lee, Won-Ja Lee, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2016;54(1):75-80.
Published online February 26, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.1.75

Abstract
PDF

This study explored epidemiological trends in trichomoniasis in Daegu, South Korea. Wet mount microscopy, PCR, and multiplex PCR were used to test for Trichomonas vaginalis in vaginal swab samples obtained from 621 women visiting 2 clinics in Daegu. Of the 621 women tested, microscopy detected T. vaginalis in 4 (0.6%) patients, PCR detected T. vaginalis in 19 (3.0%) patients, and multiplex PCR detected T. vaginalis in 12 (1.9%) patients. Testing via PCR demonstrated high sensitivity and high negative predictive value for T. vaginalis. Among the 19 women who tested positive for T. vaginalis according to PCR, 94.7% (18/19) reported vaginal signs and symptoms. Notably, more than 50% of T. vaginalis infections occurred in females younger than 30 years old, and 58% were unmarried. Multiplex PCR, which simultaneously detects pathogens from various sexually transmitted infections, revealed that 91.7% (11/12) of patients were infected with 2 or more pathogens. Mycoplasma hominis was the most prevalent co-infection pathogen with T. vaginalis, followed by Ureaplasma urealyticum and Chlamydia trachomatis. Our results indicate that PCR and multiplex PCR are the most sensitive tools for T. vaginalis diagnosis, rather than microscopy which has been routinely used to detect T. vaginalis infections in South Korea. Therefore, clinicians should take note of the high prevalence of T. vaginalis infections among adolescent and young women in order to prevent persistent infection and transmission of this disease.

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Russian Journal of Clinical Dermatology and Venereology.2024; 23(4): 377. CrossRef
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Nagarajan L. Surya, Thangamani Suji, Santhosh Rani, Irene Dorathy, Shantidani Minz, Rani Diana Sahni
BMC Infectious Diseases.2024;[Epub] CrossRef
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Mohammad Saleh Bahreini, Samaneh Sedghi, Yalda Badalzadeh, Mohammad Hossein Motazedian, Manouchehr Shirani, Sareh Sami Jahromi, Aref Teimouri, Mahmoud Agholi, Qasem Asgari
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Turkish Journal of Parasitology.2022; 46(3): 167. CrossRef
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Lauren Hernández-Buelvas, Milena Camargo, Ricardo Sánchez, Manuel Elkin Patarroyo, Manuel Alfonso Patarroyo
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Parasitology Research.2020; 119(6): 1915. CrossRef
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Sun Huh
Journal of the Korean Medical Association.2019; 62(8): 437. CrossRef
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Pati Aji Achdiat, Reiva Farah Dwiyana, Vina Feriza, Rasmia Rowawi, Rendy Ariezal Effendi, Oki Suwarsa, Hendra Gunawan
Indonesian Journal of Tropical and Infectious Disease.2019; 7(4): 57. CrossRef
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Seung-Ryong Kim, Jung-Hyun Kim, Na-Yeong Gu, Yong-Suk Kim, Yeon-Chul Hong, Jae-Sook Ryu
The Korean Journal of Parasitology.2016; 54(2): 187. CrossRef
Loop-Mediated Isothermal Amplification Targeting Actin DNA of Trichomonas vaginalis
Youn-Kyoung Goo, Won-Sik Shin, Hye-Won Yang, So-Young Joo, Su-Min Song, Jae-Sook Ryu, Hyun-Hee Kong, Won-Ki Lee, Dong-Il Chung, Yeonchul Hong
The Korean Journal of Parasitology.2016; 54(3): 329. CrossRef
Epidemiology of Trichomoniasis in South Korea and Increasing Trend in Incidence, Health Insurance Review and Assessment 2009-2014
So-Young Joo, Youn-Kyoung Goo, Jae-Sook Ryu, Sang-Eun Lee, Won Kee Lee, Dong-Il Chung, Yeonchul Hong, Zhefeng Meng
PLOS ONE.2016; 11(12): e0167938. CrossRef

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Prevalence of Trichomonas vaginalis by PCR in Men Attending a Primary Care Urology Clinic in South Korea: Jun-Hyeok Seo, Hye-Won Yang, So-Young Joo, Su-Min Song, Yu-Ran Lee, Jae-Sook Ryu, Eun Sang Yoo, Won Kee Lee, Hyun-Hee Kong, Sang-Eun Lee, Won-Ja Lee, Youn-Kyoung Goo, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2014;52(5):551-555.
Published online October 22, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.5.551

Abstract
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Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic nongonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.

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Systematic review and meta-analysis of the global prevalence and infection risk factors of Trichomonas vaginalis
Wenjie Tian, Yuhua Li, Yani Zhang, Yiming Zhang, Yiran Qin, Yalin Han, Dongxian Li, Shuai Wang, Zhenke Yang, Xiaowei Tian, Xuefang Mei, Zhenchao Zhang
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Prevalence and Genotype of Trichomonas vaginalis among Men in Xinxiang City, Henan Province, China
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Journal of Tropical Medicine.2023; 2023: 1. CrossRef
Inflammatory response to Trichomonas vaginalis in the pathogenesis of prostatitis and benign prostatic hyperplasia
Ik-Hwan Han, Jung-Hyun Kim, Jae-Sook Ryu
Parasites, Hosts and Diseases.2023; 61(1): 2. CrossRef
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International Journal of Dermatology.2022;[Epub] CrossRef
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Comparison of Seropositivity to Trichomonas vaginalis between Men with Prostatic Tumor and Normal Men
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Comparison of Two PCR Assays for Trichomonas vaginalis
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Original Article

Down-Regulation of Cellulose Synthase Inhibits the Formation of Endocysts in Acanthamoeba: Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong; Korean J Parasitol 2014;52(2):131-135.
Published online April 18, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.2.131

Abstract
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Acanthamoeba cysts are resistant to unfavorable physiological conditions and various disinfectants. Acanthamoeba cysts have 2 walls containing various sugar moieties, and in particular, one third of the inner wall is composed of cellulose. In this study, it has been shown that down-regulation of cellulose synthase by small interfering RNA (siRNA) significantly inhibits the formation of mature Acanthamoeba castellanii cysts. Calcofluor white staining and transmission electron microscopy revealed that siRNA transfected amoeba failed to form an inner wall during encystation and thus are likely to be more vulnerable. In addition, the expression of xylose isomerase, which is involved in cyst wall formation, was not altered in cellulose synthase down-regulated amoeba, indicating that cellulose synthase is a crucial factor for inner wall formation by Acanthamoeba during encystation.

Citations

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Mathew Choaji, Ascel Samba-Louaka, Zineb Fechtali-Moute, Willy Aucher, Sébastien Pomel
Pathogens.2025; 14(3): 268. CrossRef
Genetic manipulation of giant viruses and their host, Acanthamoeba castellanii
Nadege Philippe, Avi Shukla, Chantal Abergel, Hugo Bisio
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Oxidase enzyme genes are differentially expressed during Acanthamoeba castellanii encystment
Christian Q. Scheckhuber, Rebeca Damián Ferrara, Jesús Gómez-Montalvo, Sutherland K. Maciver, Alvaro de Obeso Fernández del Valle
Parasitology Research.2024;[Epub] CrossRef
Efficacy of propolis extract and eye drop solutions to suppress encystation and excystation of Acanthamoeba triangularis WU19001-T4 genotype
Suthinee Sangkanu, Abolghasem Siyadatpanah, Roghayeh Norouzi, Julalak Chuprom, Watcharapong Mitsuwan, Sirirat Surinkaew, Rachasak Boonhok, Alok K. Paul, Tooba Mahboob, Imran Sama-ae, Sonia M. R. Oliveira, Tajudeen O. Jimoh, Maria de Lourdes Pereira, Polra
PeerJ.2024; 12: e16937. CrossRef
Human Conjunctival Transcriptome in Acanthamoeba Keratitis: An Exploratory Study
Gerami D. Seitzman, Jeremy D. Keenan, Thomas M. Lietman, Kevin Ruder, Lina Zhong, Cindi Chen, YuHeng Liu, Danny Yu, Thomas Abraham, Armin Hinterwirth, Thuy Doan
Cornea.2024; 43(10): 1272. CrossRef
Evolution of giant pandoravirus revealed by CRISPR/Cas9
Hugo Bisio, Matthieu Legendre, Claire Giry, Nadege Philippe, Jean-Marie Alempic, Sandra Jeudy, Chantal Abergel
Nature Communications.2023;[Epub] CrossRef
Biological characteristics and pathogenicity of Acanthamoeba
Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
Frontiers in Microbiology.2023;[Epub] CrossRef
Anti-Acanthamoeba activity of a semi-synthetic mangostin derivative and its ability in removal of Acanthamoeba triangularis WU19001 on contact lens
Julalak Chuprom, Suthinee Sangkanu, Watcharapong Mitsuwan, Rachasak Boonhok, Wilawan Mahabusarakam, L. Ravithej Singh, Ekachai Dumkliang, Kritamorn Jitrangsri, Alok K. Paul, Sirirat Surinkaew, Polrat Wilairatana, Maria de Lourdes Pereira, Mohammed Rahmatu
PeerJ.2022; 10: e14468. CrossRef
Peganum harmala Extract Has Antiamoebic Activity to Acanthamoeba triangularis Trophozoites and Changes Expression of Autophagy-Related Genes
Rachasak Boonhok, Suthinee Sangkanu, Julalak Chuprom, Mayuna Srisuphanunt, Roghayeh Norouzi, Abolghasem Siyadatpanah, Farzaneh Mirzaei, Watcharapong Mitsuwan, Sueptrakool Wisessombat, Maria de Lourdes Pereira, Mohammed Rahmatullah, Polrat Wilairatana, Chr
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The role of the Acanthamoeba castellanii Sir2-like protein in the growth and encystation of Acanthamoeba
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MohammadHossein Feiz Haddad, Azar Shokri, Habib Habibpour, SeyedeManizhe Heidar Neiadi
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Pauline Schaap, Christina Schilde
Microbiology.2018; 164(5): 727. CrossRef
Molecular and biochemical characterization of key enzymes in the cysteine and serine metabolic pathways of Acanthamoeba castellanii
Duo Wu, Meng Feng, Zhi-xin Wang, Ke Qiao, Hiroshi Tachibana, Xun-jia Cheng
Parasites & Vectors.2018;[Epub] CrossRef
DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation
Eun-Kyung Moon, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
The Korean Journal of Parasitology.2017; 55(2): 115. CrossRef
In-vitro development of an effective treatment for Acanthamoeba keratitis
Á. Ortillés, J. Belloc, E. Rubio, M.T. Fernández, M. Benito, J.Á. Cristóbal, B. Calvo, P. Goñi
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Sahreena Lakhundi, Ruqaiyyah Siddiqui, Naveed Ahmed Khan
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An update onAcanthamoebakeratitis: diagnosis, pathogenesis and treatment
Jacob Lorenzo-Morales, Naveed A. Khan, Julia Walochnik
Parasite.2015; 22: 10. CrossRef
Potential Value of Cellulose Synthesis Inhibitors Combined With PHMB in the Treatment of Acanthamoeba Keratitis
Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong
Cornea.2015; 34(12): 1593. CrossRef

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Case Report

Heavy Hymenolepis nana Infection Possibly Through Organic Foods: Report of a Case: Bong Jin Kim, Kyung Seob Song, Hyun-Hee Kong, Hee-Jae Cha, Meesun Ock; Korean J Parasitol 2014;52(1):85-87.
Published online February 19, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.1.85

Abstract
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We encountered a patient with heavy Hymenolepis nana infection. The patient was a 44-year-old Korean man who had suffered from chronic hepatitis (type B) for 15 years. A large number of H. nana adult worms were found during colonoscopy that was performed as a part of routine health screening. The parasites were scattered throughout the colon, as well as in the terminal ileum, although the patient was immunocompetent. Based on this study, colonoscopy may be helpful for diagnosis of asymptomatic H. nana infections.

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A neglected zoonotic helminth—Hymenolepis nana, fatal in the pediatric population—an incidental finding
Smita Mohod, Swati Bhise, Sonakshi Dwivedi, Aditya Chaturvedi
Indian Journal of Pathology and Microbiology.2025; 68(4): 839. CrossRef
An Unexpected Encounter During a Screening Colonoscopy in a Medical Tourist: A Diagnosis of Hymenolepis nana Infection
Bosung Park, Hye Won Park, Hyun Jung Park, Eun Jeong Won, Heungsup Sung, Mi-Na Kim
Annals of Laboratory Medicine.2024; 44(6): 608. CrossRef
Zoonotic importance of ecto- and endoparasites in rodents in Malaysia: a review
Firdaus Mohd-Qawiem, Md Isa Nur-Mahiza, Nur Indah Ahmad, Abd Rahaman Yasmin, Saulol Hamid Nur-Fazila
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Tunable Polymeric Mixed Micellar Nanoassemblies of Lutrol F127/Gelucire 44/14 for Oral Delivery of Praziquantel: A Promising Nanovector against Hymenolepis nana in Experimentally-Infected Rats
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Original Articles

Identification of Atg8 Isoform in Encysting Acanthamoeba: Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2013;51(5):497-502.
Published online October 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.5.497

Abstract
PDF

Autophagy-related protein 8 (Atg8) is an essential component of autophagy formation and encystment of cyst-forming parasites, and some protozoa, such as, Acanthamoeba, Entamoeba, and Dictyostelium, have been reported to possess a type of Atg8. In this study, an isoform of Atg8 was identified and characterized in Acanthamoeba castellanii (AcAtg8b). AcAtg8b protein was found to encode 132 amino acids and to be longer than AcAtg8 protein, which encoded 117 amino acids. Real-time PCR analysis showed high expression levels of AcAtg8b and AcAtg8 during encystation. Fluorescence microscopy demonstrated that AcAtg8b is involved in the formation of the autophagosomal membrane. Chemically synthesized siRNA against AcAtg8b reduced the encystation efficiency of Acanthamoeba, confirming that AcAtg8b, like AcAtg8, is an essential component of cyst formation in Acanthamoeba. Our findings suggest that Acanthamoeba has doubled the number of Atg8 gene copies to ensure the successful encystation for survival when 1 copy is lost. These 2 types of Atg8 identified in Acanthamoeba provide important information regarding autophagy formation, encystation mechanism, and survival of primitive, cyst-forming protozoan parasites.

Citations

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Encystment and Excystment Processes in Acanthamoeba castellanii: An Emphasis on Cellulose Involvement
Mathew Choaji, Ascel Samba-Louaka, Zineb Fechtali-Moute, Willy Aucher, Sébastien Pomel
Pathogens.2025; 14(3): 268. CrossRef
Efficacy of propolis extract and eye drop solutions to suppress encystation and excystation of Acanthamoeba triangularis WU19001-T4 genotype
Suthinee Sangkanu, Abolghasem Siyadatpanah, Roghayeh Norouzi, Julalak Chuprom, Watcharapong Mitsuwan, Sirirat Surinkaew, Rachasak Boonhok, Alok K. Paul, Tooba Mahboob, Imran Sama-ae, Sonia M. R. Oliveira, Tajudeen O. Jimoh, Maria de Lourdes Pereira, Polra
PeerJ.2024; 12: e16937. CrossRef
Ac-HSP20 regulates autophagy and promotes the encystation of Acanthamoeba castellanii by inhibiting the PI3K/AKT/mTOR signaling pathway
Siyao Guo, Di Liu, Xi Wan, Dingrui Guo, Meiyu Zheng, Wenyu Zheng, Xianmin Feng
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Yuehua Wang, Linzhe Jiang, Yitong Zhao, Xiaohong Ju, Le Wang, Liang Jin, Ryan D. Fine, Mingguang Li
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Zineb Fechtali-Moute, Philippe M. Loiseau, Sébastien Pomel
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Maryia Karpiyevich, Katerina Artavanis-Tsakonas
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Pauline Schaap, Christina Schilde
Microbiology.2018; 164(5): 727. CrossRef
Autophagy protein 12 plays an essential role in Acanthamoeba encystation
So-Hee Kim, Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Hyun-Hee Kong
Experimental Parasitology.2015; 159: 46. CrossRef
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Eun-Kyung Moon, So-Hee Kim, Yeonchul Hong, Dong-Il Chung, Youn-Kyoung Goo, Hyun-Hee Kong
Antimicrobial Agents and Chemotherapy.2015; 59(7): 4020. CrossRef

9,819 View
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Crossref

Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Acanthamoeba: Hye-Won Yang, Yu-Ran Lee, Noboru Inoue, Bijay Kumar Jha, Dinzouna-Boutamba Sylvatrie Danne, Hong-Kyun Kim, Junhun Lee, Youn-Kyoung Goo, Hyun-Hee Kong, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2013;51(3):269-277.
Published online June 30, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.3.269

Abstract
PDF

Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner.

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Brief Communication

Short-Cut Pathway to Synthesize Cellulose of Encysting Acanthamoeba: Eun-Kyung Moon, Hyun-Hee Kong; Korean J Parasitol 2012;50(4):361-364.
Published online November 26, 2012; DOI: https://doi.org/10.3347/kjp.2012.50.4.361

Abstract
PDF

The mature cyst of Acanthamoeba is highly resistant to various antibiotics and therapeutic agents. Cyst wall of Acanthamoeba are composed of cellulose, acid-resistant proteins, lipids, and unidentified materials. Because cellulose is one of the primary components of the inner cyst wall, cellulose synthesis is essential to the process of cyst formation in Acanthamoeba. In this study, we hypothesized the key and short-step process in synthesis of cellulose from glycogen in encysting Acanthamoeba castellanii, and confirmed it by comparing the expression pattern of enzymes involving glycogenolysis and cellulose synthesis. The genes of 3 enzymes, glycogen phosphorylase, UDP-glucose pyrophosphorylase, and cellulose synthase, which are involved in the cellulose synthesis, were expressed high at the 1st and 2nd day of encystation. However, the phosphoglucomutase that facilitates the interconversion of glucose 1-phosphate and glucose 6-phosphate expressed low during encystation. This report identified the short-cut pathway of cellulose synthesis required for construction of the cyst wall during the encystation process in Acanthamoeba. This study provides important information to understand cyst wall formation in encysting Acanthamoeba.

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Down-Regulation of Cellulose Synthase Inhibits the Formation of Endocysts in Acanthamoeba
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Original Articles

Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites of Acanthamoeba castellanii: Eun-Kyung Moon, Ying-Hua Xuan, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong; Korean J Parasitol 2011;49(4):341-347.
Published online December 16, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.4.341

Abstract
PDF

Acanthamoeba infection is difficult to treat because of the resistance property of Acanthamoeba cyst against the host immune system, diverse antibiotics, and therapeutic agents. To identify encystation mediating factors of Acanthamoeba, we compared the transcription profile between cysts and trophozoites using microarray analysis. The DNA chip was composed of 12,544 genes based on expressed sequence tag (EST) from an Acanthamoeba ESTs database (DB) constructed in our laboratory, genetic information of Acanthamoeba from TBest DB, and all of Acanthamoeba related genes registered in the NCBI. Microarray analysis indicated that 701 genes showed higher expression than 2 folds in cysts than in trophozoites, and 859 genes were less expressed in cysts than in trophozoites. The results of real-time PCR analysis of randomly selected 9 genes of which expression was increased during cyst formation were coincided well with the microarray results. Eukaryotic orthologous groups (KOG) analysis showed an increment in T article (signal transduction mechanisms) and O article (posttranslational modification, protein turnover, and chaperones) whereas significant decrement of C article (energy production and conversion) during cyst formation. Especially, cystein proteinases showed high expression changes (282 folds) with significant increases in real-time PCR, suggesting a pivotal role of this proteinase in the cyst formation of Acanthamoeba. The present study provides important clues for the identification and characterization of encystation mediating factors of Acanthamoeba.

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The role of the Acanthamoeba castellanii Sir2-like protein in the growth and encystation of Acanthamoeba
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Cytopathic Change and Inflammatory Response of Human Corneal Epithelial Cells Induced by Acanthamoeba castellanii Trophozoites and Cysts
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DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation
Eun-Kyung Moon, Yeonchul Hong, Hae-Ahm Lee, Fu-Shi Quan, Hyun-Hee Kong
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Identification and Characterization of Protein Arginine Methyltransferase 1 in Acanthamoeba castellanii
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Essential Role for an M17 Leucine Aminopeptidase in Encystation of Acanthamoeba castellanii
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The Korean Journal of Parasitology.2014; 52(2): 131. CrossRef
Microarray and KOG analysis of Acanthamoeba healyi genes up-regulated by mouse-brain passage
Eun-Kyung Moon, Ying-Hua Xuan, Hyun-Hee Kong
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Proteomic profiling of the infective trophozoite stage of Acanthamoeba polyphaga
Karin Silva Caumo, Karina Mariante Monteiro, Thiely Rodrigues Ott, Vinicius José Maschio, Glauber Wagner, Henrique Bunselmeyer Ferreira, Marilise Brittes Rott
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Cysteine Protease Inhibitor (AcStefin) Is Required for Complete Cyst Formation of Acanthamoeba
Jung-Yub Lee, Su-Min Song, Eun-Kyung Moon, Yu-Ran Lee, Bijay Kumar Jha, Dinzouna-Boutamba Sylvatrie Danne, Hee-Jae Cha, Hak Sun Yu, Hyun-Hee Kong, Dong-Il Chung, Yeonchul Hong
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Cysteine protease involving in autophagosomal degradation of mitochondria during encystation of Acanthamoeba
Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Hyun-Hee Kong
Molecular and Biochemical Parasitology.2012; 185(2): 121. CrossRef
Short-Cut Pathway to Synthesize Cellulose of Encysting Acanthamoeba
Eun-Kyung Moon, Hyun-Hee Kong
The Korean Journal of Parasitology.2012; 50(4): 361. CrossRef
Protein kinase C signaling molecules regulate encystation of Acanthamoeba
Eun-Kyung Moon, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong
Experimental Parasitology.2012; 132(4): 524. CrossRef

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Expressed Sequence Tag Analysis of the Erythrocytic Stage of Plasmodium berghei: Ji-Woong Seok, Yong-Seok Lee, Eun-Kyung Moon, Jung-Yub Lee, Bijay Kumar Jha, Hyun-Hee Kong, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2011;49(3):221-228.
Published online September 30, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.3.221

Abstract
PDF

Rodent malaria parasites, such as Plasmodium berghei, are practical and useful model organisms for human malaria research because of their analogies to the human malaria in terms of structure, physiology, and life cycle. Exploiting the available genetic sequence information, we constructed a cDNA library from the erythrocytic stages of P. berghei and analyzed the expressed sequence tag (EST). A total of 10,040 ESTs were generated and assembled into 2,462 clusters. These EST clusters were compared against public protein databases and 48 putative new transcripts, most of which were hypothetical proteins with unknown function, were identified. Genes encoding ribosomal or membrane proteins and purine nucleotide phosphorylases were highly abundant clusters in P. berghei. Protein domain analyses and the Gene Ontology functional categorization revealed translation/protein folding, metabolism, protein degradation, and multiple family of variant antigens to be mainly prevalent. The presently-collected ESTs and its bioinformatic analysis will be useful resources to identify for drug target and vaccine candidates and validate gene predictions of P. berghei.

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59 Download

Atg3-Mediated Lipidation of Atg8 Is Involved in Encystation of Acanthamoeba: Eun-Kyung Moon, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong; Korean J Parasitol 2011;49(2):103-108.
Published online June 14, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.2.103

Abstract
PDF

Autophagy is a catabolic process involved in the degradation of a cell's own components for cell growth, development, homeostasis, and the recycling of cellular products. Autophagosome is an essential component in the protozoan parasite during differentiation and encystation. The present study identified and characterized autophagy-related protein (Atg) 3, a member of Atg8 conjugation system, in Acanthamoeba castellanii (AcAtg3). AcAtg3 encoding a 304 amino acid protein showed high similarity with the catalytic cysteine site of other E2 like enzymes of ubiquitin system. Predicted 3D structure of AcAtg3 revealed a hammer-like shape, which is the characteristic structure of E2-like enzymes. The expression level of AcAtg3 did not increase during encystation. However, the formation of mature cysts was significantly reduced in Atg3-siRNA transfected cells in which the production of Atg8-phosphatidylethanolamine conjugate was inhibited. Fluorescent microscopic analysis revealed that dispersed AcAtg3-EGFP fusion protein gathered around autophagosomal membranes during encystation. These results provide important information for understanding autophagic machinery through the lipidation reaction mediated by Atg3 in Acanthamoeba.

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Cysteine protease involving in autophagosomal degradation of mitochondria during encystation of Acanthamoeba
Eun-Kyung Moon, Yeonchul Hong, Dong-Il Chung, Hyun-Hee Kong
Molecular and Biochemical Parasitology.2012; 185(2): 121. CrossRef
Protein kinase C signaling molecules regulate encystation of Acanthamoeba
Eun-Kyung Moon, Dong-Il Chung, Yeonchul Hong, Hyun-Hee Kong
Experimental Parasitology.2012; 132(4): 524. CrossRef
Microarray Analysis of Differentially Expressed Genes between Cysts and Trophozoites ofAcanthamoeba castellanii
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The Korean Journal of Parasitology.2011; 49(4): 341. CrossRef

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Book Review

Acanthamoeba: Biology and Pathogenesis: Hyun-Hee Kong; Korean J Parasitol 2010;48(2):187-187.
Published online June 17, 2010; DOI: https://doi.org/10.3347/kjp.2010.48.2.187

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Original Articles

Construction of EST Database for Comparative Gene Studies of Acanthamoeba: Eun-Kyung Moon, Joung-Ok Kim, Ying-Hua Xuan, Young-Sun Yun, Se Won Kang, Yong Seok Lee, Tae-In Ahn, Yeon-Chul Hong, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2009;47(2):103-107.
Published online May 26, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.2.103

Abstract
PDF

The genus Acanthamoeba can cause severe infections such as granulomatous amebic encephalitis and amebic keratitis in humans. However, little genomic information of Acanthamoeba has been reported. Here, we constructed Acanthamoeba expressed sequence tags (EST) database (Acanthamoeba EST DB) derived from our 4 kinds of Acanthamoeba cDNA library. The Acanthamoeba EST DB contains 3,897 EST generated from amebae under various conditions of long term in vitro culture, mouse brain passage, or encystation, and downloaded data of Acanthamoeba from National Center for Biotechnology Information (NCBI) and Taxonomically Broad EST Database (TBestDB). The almost reported cDNA/genomic sequences of Acanthamoeba provide stand alone BLAST system with nucleotide (BLAST NT) and amino acid (BLAST AA) sequence database. In BLAST results, each gene links for the significant information including sequence data, gene orthology annotations, relevant references, and a BlastX result. This is the first attempt for construction of Acanthamoeba database with genes expressed in diverse conditions. These data were integrated into a database (http://www.amoeba.or.kr).

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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
PDF

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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Brief Communication

Differentially expressed genes of Acanthamoeba castellanii during encystation: Eun-Kyung Moon, Dong-Il Chung, Yeon-Chul Hong, Hyun-Hee Kong; Korean J Parasitol 2007;45(4):283-285.
Published online December 20, 2007; DOI: https://doi.org/10.3347/kjp.2007.45.4.283

Abstract
PDF

To examine the expressed gene profile during encystation of Acanthamoeba castellanii Castellani, we used differentially expressed gene (DGE) screening by RT-PCR with 20 sets of random primers. From this analysis, we found that approximately 16 genes showed upregulation during encystation. We chose 6 genes, which had relatively higher expression levels, for further investigation. Based on homology search in database, DEG2 showed 55% of similarity with xylose isomerase, DEG9 showed 37% of similarity with Na P-type ATPase, and DEG14 showed 77% of similarity with subtilisin-like serine proteinase. DEG3 and DEG26 were identified as hypothetical proteins and DEG25 exhibited no significant similarity to any known protein. Encystation of Acanthamoeba has been suggested to be a process to resist adverse environmental or nutritional conditions. Further characterization studies of these genes may provide us with more information on the encystation mechanism of Acanthamoeba.

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Original Articles

Natural occurrence of Mycobacterium as an endosymbiont of Acanthamoeba isolated from a contact lens storage case: Hak Sun Yu, Hae Jin Jeong, Yeon-Chul Hong, Seong-Yong Seol, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2007;45(1):11-18.
Published online March 20, 2007; DOI: https://doi.org/10.3347/kjp.2007.45.1.11

Abstract
PDF

Recent in vitro studies have revealed that a certain Mycobacterium can survive and multiply within free-living amoebae. It is believed that protozoans function as host cells for the intracellular replication and evasion of Mycobacterium spp. under harmful conditions. In this study, we describe the isolation and characterization of a bacterium naturally observed within an amoeba isolate acquired from a contact lens storage case. The bacterium multiplied within Acanthamoeba, but exerted no cytopathic effects on the amoeba during a 6-year amoebic culture. Trasnmission electron microscopy showed that the bacteria were randomly distributed within the cytoplasm of trophozoites and cysts of Acanthamoeba. On the basis of the results of 18S rRNA gene analysis, the amoeba was identified as A. lugdunensis. A 16S rRNA gene analysis placed this bacterium within the genus Mycobacterium. The bacterium evidenced positive reactivity for acid-fast and fluorescent acid-fast stains. The bacterium was capable of growth on the Middlebrook 7H11-Mycobacterium-specific agar. The identification and characterization of bacterial endosymbionts of free-living protozoa bears significant implications for our understanding of the ecology and the identification of other atypical mycobacterial pathogens.

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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
PDF

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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Molecular and biochemical characterization of a novel actin bundling protein in Acanthamoeba: Joanna It-itan Alafag, Eun-Kyung Moon, Yeon-Chul Hong, Dong-Il Chung, Hyun-Hee Kong; Korean J Parasitol 2006;44(4):331-341.
Published online December 20, 2006; DOI: https://doi.org/10.3347/kjp.2006.44.4.331

Abstract
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Actin binding proteins play key roles in cell structure and movement particularly as regulators of the assembly, stability and localization of actin filaments in the cytoplasm. In the present study, a cDNA clone encoding an actin bundling protein named as AhABP was isolated from Acanthamoeba healyi, a causative agent of granulomatous amebic encephalitis. This clone exhibited high similarity with genes of Physarum polycephalum and Dictyostelium discoideum, which encode actin bundling proteins. Domain search analysis revealed the presence of essential conserved regions, i.e., an active actin binding site and 2 putative calcium binding EF-hands. Transfected amoeba cells demonstrated that AhABP is primarily localized in phagocytic cups, peripheral edges, pseudopods, and in cortical cytoplasm where actins are most abundant. Moreover, AhABP after the deletion of essential regions formed ellipsoidal inclusions within transfected cells. High-speed co-sedimentation assays revealed that AhABP directly interacted with actin in the presence of up to 10 ?M of calcium. Under the electron microscope, thick parallel bundles were formed by full length AhABP, in contrast to the thin actin bundles formed by constructs with deletion sites. In the light of these results, we conclude that AhABP is a novel actin bundling protein that is importantly associated with actin filaments in the cytoplasm.

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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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Eun Chul Kim, Man Soo Kim
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F R S Carvalho, A S Foronda, M J Mannis, A L Höfling-Lima, R Belfort, Denise de Freitas
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Hyun Hee Kong
The Korean Journal of Parasitology.2009; 47(Suppl): S21. CrossRef
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Molecular and biochemical characterization of hemoglobinase, a cysteine proteinase, in Paragonimus westermani: Joon-Hyuck Choi, Jae-Hyuk Lee, Hak-Sun Yu, Hae-Jin Jeong, Jin Kim, Yeon-Chul Hong, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2006;44(3):187-196.
Published online September 20, 2006; DOI: https://doi.org/10.3347/kjp.2006.44.3.187

Abstract
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The mammalian trematode Paragonimus westermani is a typical digenetic parasite, which can cause paragonimiasis in humans. Host tissues and blood cells are important sources of nutrients for development, growth and reproduction of P. westermani. In this study, a cDNA clone encoding a 47 kDa hemoglobinase of P. westermani was characterized by sequencing analysis, and its localization was investigated immunohistochemically. The phylogenetic tree prepared based on the hemoglobinase gene showed high homology with hemoglobinases of Fasciola hepatica and Schistosoma spp. Moreover, recombinant P. westermani hemoglobinase degradaded human hemoglobin at acidic pH (from 3.0 to 5.5) and its activity was almost completely inhibited by E-64, a cysteine proteinase inhibitor. Immunohistochemical studies showed that P. westermani hemoglobinase was localized in the epithelium of the adult worm intestine implying that the protein has a specific function. These observations suggest that hemoglobinase may act as a digestive enzyme for acquisition of nutrients from host hemoglobin. Further investigations may provide insights into hemoglobin catabolism in P. westermani.

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Genetic diversity of Acanthamoeba isolates from ocean sediments: Hua Liu, Young-Ran Ha, Sung-Tae Lee, Yean-Chul Hong, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2006;44(2):117-125.
Published online June 20, 2006; DOI: https://doi.org/10.3347/kjp.2006.44.2.117

Abstract
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Genetic diversity of 18 Acanthamoeba isolates from ocean sediments was evaluated by comparing mitochondrial (mt) DNA RFLP, 18S rDNA sequences and by examining their cytopathic effects on human corneal epithelial cells versus reference strains. All isolates belonged to morphologic group II. Total of 16 restriction phenotypes of mtDNA from 18 isolates demonstrated the genetic diversity of Acanthamoeba in ocean sediments. Phylogenetic analysis using 18s rDNA sequences revealed that the 18 isolates were distinct from morphological groups I and III. Fifteen isolates showed close relatedness with 17 clinical isolates and A. castellanii Castellani and formed a lineage equivalent to T4 genotype of Byers' group. Two reference strains from ocean sediment, A. hatchetti BH-2 and A. griffini S-7 clustered unequivocally with these 15 isolates. Diversity among isolates was also evident from their cytopathic effects on human corneal cells. This is the first time describing Acanthamoeba diversity in ocean sediments in Korea.

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Evaluation of taxonomic validity of four species of Acanthamoeba: A. divionensis, A. paradivionensis, A. mauritaniensis, and A. rhysodes, inferred from molecular analyses: Hua Liu, Eun-Kyung Moon, Hak-Sun Yu, Hae-Jin Jeong, Yeon-Chul Hong, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2005;43(1):7-13.
Published online March 20, 2005; DOI: https://doi.org/10.3347/kjp.2005.43.1.7

Abstract
PDF

The taxonomy of Acanthamoeba spp., an amphizoic amoeba which causes granulomatous amoebic encephalitis and chronic amoebic keratitis, has been revised many times. The taxonomic validity of some species has yet to be assessed. In this paper, we analyzed the morphological characteristics, nuclear 18s rDNA and mitochondrial 16s rDNA sequences and the Mt DNA RFLP of the type strains of four Acanthamoeba species, which had been previously designated as A. divionensis, A. parasidionensis, A. mauritaniensis, and A. rhysodes. The four isolates revealed characteristic group II morphology. They exhibited 18S rDNA sequence differences of 0.2-1.1% with each other, but more than 2% difference from the other compared reference strains. Four isolates formed a different clade from that of A. castellanii Castellani and the other strains in morphological group II on the phylogenetic tree. In light of these results, A. paradivionensis, A. divionensis, and A. mauritaniensis should be regarded as synonyms for A. rhysodes.

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Stas Malavin, Lyubov Shmakova
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Daniele Corsaro
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Md Moshiur Rahman, Kenji Yagita, Akira Kobayashi, Yosaburo Oikawa, Amjad I.A. Hussein, Takahiro Matsumura, Masaharu Tokoro
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The Korean Journal of Parasitology.2008; 46(3): 157. CrossRef
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Hatice Ertabaklar, Meral Türk, Volkan Dayanir, Sema Ertuğ, Julia Walochnik
Parasitology Research.2007; 100(2): 241. CrossRef

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Brief Communication

Infection status of intestinal parasites in children living in residential institutions in Metro Manila, the Philippines: Eleonor T Baldo, Vicente Y Belizario, Winifreda U De Leon, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2004;42(2):67-70.
Published online June 20, 2004; DOI: https://doi.org/10.3347/kjp.2004.42.2.67

Abstract
PDF

A small scale survey was performed to know the infection status of intestinal parasite in children of the residential institutions and street communities in Metro Manila, Philippines. A total of 284 stool samples from 11 institutions and 3 street communities was examined by the formalin-ether concentration method. The scotch tape anal swab was adapted to 121 children to investigate the infection status of Enterovius vermicularis. It was found out that 62.0% of the children examined were positive for one or more intestinal parasites. Multiple infections were observed in 34.2% of the children. Among 172 children who gave detail information, the prevalence for Ascaris lumbricoides, Trichuris trichiura, and hookworm was 36.0%, 44.8%, and 7.0% respectively. Of the children examined, 47.7% were found to be harboring parasitic protozoans such as Entamoeba histolytica, Giardia lamblia, and Blastocystis hominis. The most prevalent of these protozoans was B. hominis with an infection rate of 40.7%. The prevalence of these infections among children living in institutions was relatively high. More efforts should be made to implement anthelminthic programs including bi-annual follow-up treatments.

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Original Article

Purification and characterization of a 33 kDa serine protease from Acanthamoeba lugdunensis KA/E2 isolated from a Korean keratitis patient: Hyo-Kyung Kim, Young-Ran Ha, Hak-Sun Yu, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2003;41(4):189-196.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.189

Abstract
PDF

In order to evaluate the possible roles of secretory proteases in the pathogenesis of amoebic keratitis, we purified and characterized a serine protease secreted by Acanthamoeba lugdunensis KA/E2, isolated from a Korean keratitis patient. The ammonium sulfate-precipitated culture supernatant of the isolate was purified by sequential chromatography on CM-Sepharose, Sephacryl S-200, and mono Q-anion exchange column. The purified 33 kDa protease had a pH optimum of 8.5 and a temperature optimum of 55℃. Phenylmethylsulfonylfluoride and 4-(2-Aminoethyl)-benzenesulfonyl-fluoride, both serine protease specific inhibitors, inhibited almost completely the activity of the 33 kDa protease whereas other classes of inhibitors did not affect its activity. The 33 kDa enzyme degraded various extracellular matrix proteins and serum proteins. Our results strongly suggest that the 33 kDa serine protease secreted from this keratopathogenic Acanthamoeba play important roles in the pathogenesis of amoebic keratitis, such as in corneal tissue invasion, immune evasion and nutrient uptake.

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Brief Communication

A human case of Echinostoma hortense (Trematoda: Echinostomatidae) infection diagnosed by gastroduodenal endoscopy in Korea: Chang-Min Cho, Won-Young Tak, Young-Oh Kweon, Sung-Kook Kim, Yong-Hwan Choi, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2003;41(2):117-120.
Published online June 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.2.117

Abstract
PDF

A human Echinostoma hortense infection was diagnosed by gastroduodenoscopy. An 81-year-old Korean male, living in Yeongcheon-shi, Gyeongsangbuk-do and with epigastric discomfort of several days duration, was subjected to upper gastrointestinal endoscopy. He was in the habit of eating fresh water fish. Two live worms were found in the duodenal bulb area and were removed using an endoscopic forcep. Based on their morphological characteristics, the worms were identified as E. hortense. The patient was treated with praziquantel 10 mg/kg as a single dose. The source of the infection in this case remains unclear, but the fresh water fish consumed, including the loach, may have been the source. This is the second case of E. hortense infection diagnosed by endoscopy in Korea.

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Original Articles

Cloning and characterization of Giardia intestinalis cyclophilin: Hak-Sun Yu, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2002;40(3):131-138.
Published online September 30, 2002; DOI: https://doi.org/10.3347/kjp.2002.40.3.131

Abstract
PDF

The cyclophilins (Cyps) are family members of proteins that exhibit peptidyl-prolyl cis-trans isomerase (PPIase, EC 5.2.1.8) activity and bind the immunosuppressive agent cyclosprin A (CsA) in varying degrees. During the process of random sequencing of a cDNA library made from Giardia intestinalis WB strain, the cyclophilin gene (gicyp1) was isolated. An open reading frame of gicyp1 gene was 576 nucleotides, which corresponded to a translation product of 176 amino acids (Gicyp1). The identity with other Cyps was about 58-71%. The 13 residues that constituted the CsA binding site of human cyclophilin were also detected in the amino acid sequence of Gicyp1, including tryptophan residue essential for the drug binding. The single copy of the gicyp1 gene was detected in the G. intestinalis chromosome by southern hybridization analysis. Recombinant Gicyp1 protein clearly accelerated the rate of cis → trans isomerization of the peptide substrate and the catalysis was completely inhibited by the addition of 0.5 ?M CsA.

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A riboprinting scheme for identification of unknown Acanthamoeba isolates at species level: Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2002;40(1):25-31.
Published online March 31, 2002; DOI: https://doi.org/10.3347/kjp.2002.40.1.25

Abstract
PDF

We describe a riboprinting scheme for identification of unknown Acanthamoeba isolates at the species level. It involved the use of PCR-RFLP of small subunit ribosomal RNA gene (riboprint) of 24 reference strains by 4 kinds of restriction enzymes. Seven strains in morphological group I and III were identified at species level with their unique sizes of PCR product and riboprint type by Rsa I. Unique RFCP of 17 strains in group II by Dde I, Taq I and Hae III were classified into: (1) four taxa that were identifiable at the species level, (2) a subgroup of 4 taxa and a pair of 2 taxa that were identical with each other, and (3) a species complex of 7 taxa assigned to A. castellanii complex that were closely related. These results were consistent with those obtained by 18s rDNA sequence analysis. This approach provides an alternative to the rDNA sequencing for rapid identification of a new clinical isolate or a large number of environmental isolates of Acanthamoeba.

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Pathogenic free-living amoebae in Korea
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Isolation and characterization of a cDNA encoding a mammalian cathepsin L-like cysteine proteinase from Acanthamoeba healyi: Yeon-Chul Hong, Mi-Yul Hwang, Ho-Cheol Yun, Hak-Sun Yu, Hyun-Hee Kong, Tai-Soon Yong, Dong-Il Chung; Korean J Parasitol 2002;40(1):17-24.
Published online March 31, 2002; DOI: https://doi.org/10.3347/kjp.2002.40.1.17

Abstract
PDF

We have cloned a cDNA encoding a cysteine proteinase of the Acanthamoeba healyi OC-3A strain isolated from the brain of a granulomatous amoebic encephalitis patient. A DNA probe for an A. healyi cDNA library screening was amplified by PCR using degenerate oligonucleotide primers designed on the basis of conserved amino acids franking the active sites of cysteine and asparagine residues that are conserved in the eukaryotic cysteine proteinases. Cysteine proteinase gene of A. healyi (AhCP1) was composed of 330 amino acids with signal sequence, a proposed pro-domain and a predicted active site made up of the catalytic residues, Cys²⁵, His¹⁵⁹, and Asn¹⁷⁵. Deduced amino acid sequence analysis indicates that AhCP1 belong to ERFNIN subfamily of C1 peptidases. By Northern blot analysis, no direct correlation was observed between AhCP1 mRNA expression and virulence of Acanthamoeba, but the gene was expressed at higher level in amoebae isolated from soil than amoeba from clinical samples. These findings raise the possibility that Ahcp1 protein may play a role in protein metabolism and digestion of phagocytosed bacteria or host tissue debris rather than in invasion of amoebae into host tissue.

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Genetic analyses of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea: Hak-Sun Yu, Kyung-Hee Choi, Hyo-Kyung Kim, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 2001;39(2):161-170.
Published online June 30, 2001; DOI: https://doi.org/10.3347/kjp.2001.39.2.161

Abstract
PDF

We conducted both the small subunit ribosomal DNA (SSU rDNA) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mitochondrial (mt) DNA RFLP analyses for a genetic characterization of Acanthamoeba isolates from contact lens storage cases of students in Seoul, Korea. Twenty-three strains of Acanthamoeba from the American Type Culture Collection and twelve clinical isolates from Korean patients were used as reference strains. Thirty-nine isolates from contact lens storage cases were classified into seven types (KA/LS1, KA/LS2, KA/LS4, KA/LS5, KA/LS7, KA/LS18, KA/LS31). Four types (KA/LS1, KA/LS2, KA/LS5, KA/LS18) including 33 isolates were regarded as A. castellanii complex by riboprints. KA/LS1 type was the most predominant (51.3%) in the present survey area, followed by KA/LS2 (20.9%), and KA/LS5 (7.7%) types. Amoebae of KA/LS1 type had the same mtDNA RFLP and riboprint patterns as KA/E2 and KA/E12 strains, clinical isolates from Korean keratitis patients. Amoebae of KA/LS2 type had the identical mtDNA RFLP patterns with A. castellanii Ma strain, a corneal isolate from an American patient as amoebae of KA/LS5 type, with KA/E3 and KA/E8 strains from other Korean keratitis patients. Amoebae of KA/LS18 type had identical patterns with JAC/E1, an ocular isolate from a Japanese patient. Three types, which remain unidentified at species level, were not corresponded with any clinical isolate in their mtDNA RFLP and riboprint patterns. Out of 39 isolates analyzed in this study, mtDNA RFLP and riboprint patterns of 33 isolates (84.6%) were identical to already known clinical isolates, and therefore, they may be regarded as potentially keratopathogenic. These results suggest that contact lens wearers in Seoul should pay more attention to hygienic maintenance of contact lens storage cases for the prevention of Acanthamoeba keratitis.

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Expressed sequence tags (ESTs) analysis of Acanthamoeba healyi: Hyun-Hee Kong, Mee-Yeul Hwang, Hyo-Kyung Kim, Dong-Il Chung; Korean J Parasitol 2001;39(2):151-160.
Published online June 30, 2001; DOI: https://doi.org/10.3347/kjp.2001.39.2.151

Abstract
PDF

Randomly selected 435 clones from Acanthamoeba healyi cDNA library were sequenced and a total of 387 expressed sequence tags (ESTs) had been generated. Based on the results of BLAST search, 130 clones (34.4%) were identified as the genes enconding surface proteins, enzymes for DNA, energy production or other metabolism, kinases and phosphatases, protease, proteins for signal transduction, structural and cytoskeletal proteins, cell cycle related proteins, transcription factors, transcription and translational machineries, and transporter proteins. Most of the genes (88.5%) are newly identified in the genus Acanthamoeba. Although 15 clones matched the genes of Acanthamoeba located in the public databases, twelve clones were actin gene which was the most frequently expressed gene in this study. These ESTs of Acanthamoeba would give valuable information to study the organism as a model system for biological investigations such as cytoskeleton or cell movement, signal transduction, transcriptional and translational regulations. These results would also provide clues to elucidate factors for pathogenesis in human granulomatous amoebic encephalitis or keratitis by Acanthamoeba.

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The Korean Journal of Parasitology.2009; 47(2): 103. CrossRef
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BRUNO DA ROCHA‐AZEVEDO, MELISSA JAMERSON, GUY A. CABRAL, FERNANDO C. SILVA‐FILHO, FRANCINE MARCIANO‐CABRAL
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Phylogenetic relationships among Acanthamoeba spp. based on PCR-RFLP analyses of mitochondrial small subunit rRNA gene: Hak-Sun Yu, Mee-Yul Hwang, Tae-Ook Kim, Ho-Cheol Yun, Tae-Ho Kim, Hyun-Hee Kong, Dong-Il Chung; Korean J Parasitol 1999;37(3):181-188.
Published online September 30, 1999; DOI: https://doi.org/10.3347/kjp.1999.37.3.181

Abstract
PDF

We investigated the value of mitochondrial small subunit rRNA gene (mt SSU rDNA) PCR-RFLP as a taxonomic tool for Acanthamoeba isolates with close interrelationships. Twenty-five isolates representing 20 species were included in the analysis. As in nuclear 18S rDNA analysis, two type strains (A. astronyxis and A. tubiashi) of morphological group 1 diverged earliest from the other strains, but the divergence between them was less than in 18S riboprinting. Acanthamoeba griffini of morphological group 2 branched between pathogenic (A. culbertsoni A-1 and A. healyi OC-3A) and nonpathogenic (A. palestinensis Reich, A. pustulosa GE-3a, A. royreba Oak Ridge, and A lenticulata PD2S) strains of morphological group 3. Among the remaining isolates of morphological group 2, the Chang strain had the identical mitochondrial riboprints as the type strain of A. hatchetti. AA2 and AA1, the type strains of A. divionensis and A. paradivionensis, respectively, had the identical riboprints as A. quina Vil3 and A. castellanii Ma. Although the branching orders of A. castellanii Neff, A. polyphaga P23, A. triangularis SH621, and A. lugdunensis L3a were different from those in 18S riboprinting analysis, the results obtained from this study generally coincided well with those from 18S riboprinting. Mitochondrial riboprinting may have an advantage over nuclear 18S rDNA riboprinting because the mt SSU rDNAs do not seem to have introns that are found in the 18S genes of Acanthamoeba and that distort phylogenetic analyses.

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