• Parasites, Hosts and Diseases
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• v.41 no.4
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• pp.181-188
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• 2003

A new species of Acanthamoeba was isolated from a freshwater fish in Korea and tentatively named Acanthamoeba sp. YM-4 (Korean isolate YM-4). The trophozoites were $11.0-23.0{\;}{\mu\textrm{m}}$ in length and had hyaline filamentous projections. Cysts were similar to those of A. culbertsoni and A. royreba, which were previously designated as Acanthamoeba group III. Acanthamoeba YM-4 can survive at $40^{\circ}C$, and its generation time was 19.6 hr, which was longer than that of A. culbertsoni. In terms of the in vitro cytotoxicity of lysates, Acanthamoeba YM-4 was weaker than A. culbertsoni, but stronger than A. polyphaga. On the basis of the mortality of experimentally infected mice, Acanthamoeba YM-4 was found to be highly virulent. The isoenzymes profile of Acanthamoeba YM-4 was similar to that of A. royreba. An anti-Acanthamoeba YM-4 monoclonal antibody, McAY7, was found to react only with Acanthamoeba YM-4, and not with A. culbertsoni. Random amplified polymorphic DNA marker analysis and RFLP analysis of mitochondrial DNA and of 18S small subunit ribosomal RNA, placed Acanthamoeba YM-4 in a separate cluster on the basis of phylogenetic distances. Thus the Acanthamoeba Korean isolate YM-4 was identified as a new species, and assigned as Acanthamoeba sohi.

• Parasites, Hosts and Diseases
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• v.43 no.2 s.134
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• pp.47-50
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• 2005

A survey was carried out from August to December 2004 in Pusan, Korea to document the presence of free-living amoeba (FLA), including the genus Acanthamoeba, in both contact lens storage cases and domestic tap water. Acanthamoeba was isolated from $5(4.2\%)$ in 120 contact lens storage cases. Four house tap water samples from residents, whose contact lens storage cases had been contaminated by Acanthamoeba, were also found to be contaminated with Acanthamoeba. Therefore, the contamination rate of FLA and Acanthamoeba in domestic tap water was investigated in order to examine the role of domestic tap water in Acanthamoeba contamination of contact lens storage cases. FLA and Acanthamoeba were identified in $97(46.8\%)\;and\;16(7.7\%)$ of the 207 domestic tap water samples, respectively. There were no significant differences between the contamination rates of FLA in tap water according to the filtration plant of origin. No FLA was detected in the tap water directly supplied by the water purification plants. Water storage tanks appear to promote FLA colonization, including Acanthamoeba, in domestic tap water. This increases the risk of Acanthamoeba contamination in contact lens storage cases as well as increasing the risk of Acanthamoeba keratitis.

• Parasites, Hosts and Diseases
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• v.42 no.3
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• pp.93-119
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• 2004

Acanthamoeba and Naegleria are widely distributed in fresh water, soil and dust throughout the world, and cause meningoencephalitis or keratoconjunctivitis in humans and other mammals. Korean isolates, namely, Naegleria sp. YM-1 and Acanthamoeba sp. YM-2, YM-3, YM-4, YM-5, YM-6 and YM-7, were collected from sewage, water puddles, a storage reservoir, the gills of a fresh water fish, and by corneal washing. These isolates were categorized into three groups based on the mortalities of infected mice namely, highly virulent (YM-4), moderately virulent (YM-2, YM-5 and YM-7) and nonpathogenic (YM-3). In addition, a new species of Acanthamoeba was isolated from a freshwater fish in Korea and tentatively named Korean isolate YM-4. The morphologic characters of its cysts were similar to those of A. culbertsoni and A. royreba, which were previously designated as Acanthamoeba group III. Based on experimentally infected mouse mortality, Acanthamoeba YM-4 was highly virulent. The isoenzymes profile of Acanthamoeba YM-4 was similar to that of A. royreba. Moreover, an anti-Acanthamoeba YM-4 monoclonal anti-body reacted only with Acanthamoeba YM-4, and not with A. culbertsoni. Random amplified polymorphic DNA marker analysis and RFLP analysis of mitochondrial DNA and of a 188 small subunit ribosomal RNA, placed Acanthamoeba YM-4 in a separate cluster based on phylogenic distances. Thus Acanthamoeba YM-4 was identified as a new species, and assigned Acanthamoeba sohi. Up to the year 2002 in Korea, two clinical cases were found to be infected with Acanthamoeba spp. These patients died of meningoencephalitis. In addition, one case of Acanthamoeba pneumonia with an immunodeficient status was reported and Acanthamoeba was detected in several cases of chronic relapsing corneal ulcer, chronic conjunctivitis, and keratitis.

• Biomedical Science Letters
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• v.17 no.3
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• pp.173-176
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• 2011

Free-living Acanthamoeba are eukaryotic protozoan organisms that are widely distributed in the air, water, etc such as environment. Acanthamoeba ingest the Escherichia coli which will replicate in cytoplasm of Acanthamoeba. Bacterial pathogenicity or virulence is one of important determinant factors to survive in free-living Acanthamoeba and otherwise Acanthamoebic pathogenicity is also an important factor for their interactions. Bacterial association with pathogenic strain of Acanthamoeba T1 and T4 was lower about two times than non-pathogenic T7. Bacterial invasion percentages into T1 were higher about three times than T7 but bacterial survival in T7 was increased as T1. The capsule-deletion mutant exhibited limited ability for invasion/uptake by and survival inside pathogenic Acanthamoeba T4. E. coli-outer membrane protein A (OmpA) decreased bacterial association with A. castellanii by about three times and it had higher effects than lipopolysaccharides (LPS). Under favorable conditions, the mutants were not survived in Acanthamoeba up to 24 h incubation. Therefore, this review will report pathogenic and non-pathogenic Acanthamoeba strains interactions with E. coli and its several mutants, i.e., capsule, OmpA and LPS.

• Biomedical Science Letters
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• v.18 no.3
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• pp.324-327
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• 2012

Acanthamoeba is a free-living amoeba that causes human infections, and recently the incidence of amoebic keratitis has increased among contact lens wearers. In order to investigate Acanthamoeba contamination of contact lens storage cases, a short survey was performed on 57 contact lens wearers, and Acanthamoeba was found in one contact lens storage case. To diagnose Acanthamoeba, the 18s small subunit ribosomal DNA (18s rDNA) gene was amplified by polymerase chain reaction (PCR), and subsequently, the isolate was identified as A. lugdunensis. This species was originally isolated from a freshwater pool in France, and was reported recently to be a cause of amoebic keratitis. This observation indicates the need for a large survey to investigate the extent of Acanthamoeba contamination, and suggests that contact lens wearers be aware of the importance of hygiene and of the implications of Acanthamoeba infection.

• Parasites, Hosts and Diseases
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• v.47 no.2
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• pp.103-107
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• 2009

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 eDNA/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).

• Biomedical Science Letters
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• v.18 no.3
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• pp.313-318
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• 2012

Acanthamoeba is an opportunistic pathogen known to cause granulomatous amoebic encephalitis and amebic keratitis. Acanthamoeba exhibits life cycle consisting of trophozoite and cyst, and the cyst is highly resistant to variable antibiotics and therapeutic agents. To understand the encystation mechanism of Acanthamoeba, the expression profiles of trophozoite and cyst were compared by gene ontology (GO) analysis. Ribosomal proteins and cytoskeletal proteins were highly expressed in trophozoite. In cyst, various protease, and signal transduction - and protein turnover - related proteins were highly expressed. These results correlated with eukaryotic orthologous groups (KOG) assignment and microarray analysis of Acanthamoeba trophozoite and cyst ESTs. The information of differential expression profiles of trophozoite and cyst would provide important clues for research on encystation mechanism of cyst forming protozoa including Acanthamoeba.

• Parasites, Hosts and Diseases
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• v.51 no.5
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• pp.497-502
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• 2013

Autophagy-related protein 8 (Atg8) is an essential component of autophagy formation and encystment of cystforming 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.

• Parasites, Hosts and Diseases
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• v.50 no.4
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• pp.361-364
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• 2012

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.

• Parasites, Hosts and Diseases
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• v.62 no.1
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• pp.139-144
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• 2024

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