Parasites, Hosts and Diseases

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Volume 41(4); December 2003

Original Articles

Acanthamoeba sohi, n. sp., a pathogenic Korean isolate YM-4 from a freshwater fish: Kyung-il Im, Ho-Joon Shin; Korean J Parasitol 2003;41(4):181-188.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.181

Abstract
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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 ?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℃, 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.

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Epidemiology of and Genetic Factors Associated with Acanthamoeba Keratitis
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Isolates of Acanthamoeba species in the marine environment in the Philippines
Samantha Nicole Layson, Cheilo Maurrice D. Alcala, Mikael Lorenzo Q. Avenido, Aleeza Erika M. Bayot, Charles Darwin C. Aclan, Joepher S. Barlis, Katrina D. Villacorta, Venice Marielle R. Abalos, Alyssa Nicole M. Maramba, Maricel D.C. Say, Alessandrea A. S
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Factors Affecting the Epidemiology ofAcanthamoebaKeratitis
Youhanna W. Ibrahim, David L. Boase, Ian A. Cree
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Role of the Nfa1 Protein in Pathogenic Naegleria fowleri Cocultured with CHO Target Cells
Su-Yeon Kang, Kyoung-Ju Song, Seok-Ryoul Jeong, Jong-Hyun Kim, Sun Park, Kyongmin Kim, Myung-Hee Kwon, Ho-Joon Shin
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Expression of the nfa1 Gene Cloned from Pathogenic Naegleria fowleri in Nonpathogenic N. gruberi Enhances Cytotoxicity against CHO Target Cells In Vitro
Seok-Ryoul Jeong, Sang-Chul Lee, Kyoung-Ju Song, Sun Park, Kyongmin Kim, Myung-Hee Kwon, Kyung-il Im, Ho-Joon Shin
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Cloning and characterization of an immunoreactive gene encoding a calcium-binding protein from Naegleria fowleri
Seok-Ryoul Jeong, Myung-Soo Cho, Sun Park, Kyongmin Hwang Kim, Kyoung-Ju Song, Kyung-Il Im, Ho-Joon Shin
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Decreasing effect of an anti-Nfa1 polyclonal antibody on the in vitro cytotoxicity of pathogenic Naegleria fowleri
Seok-Ryoul Jeong, Su-Yeon Kang, Sang-Chul Lee, Kyoung-Ju Song, Kyung-il Im, Ho-Joon Shin
The Korean Journal of Parasitology.2004; 42(1): 35. CrossRef
Pathogenic free-living amoebae in Korea
Ho-Joon Shin, Kyung-il Im
The Korean Journal of Parasitology.2004; 42(3): 93. CrossRef

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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
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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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Viability of preserved Cryptosporidium baileyi oocysts: Chan-Gu Surl, Se-Min Kim, Hyeon-Cheol Kim; Korean J Parasitol 2003;41(4):197-201.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.197

Abstract
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The present study was undertaken to determine the viability and infectivity of oocysts of Cryptosporidium baileyi that had been stored from 1 to 40 months at 4℃ preserved in 2.5% potassium dichromate solution. Oocysts of C. baileyi were purified from the feces of experimentally infected chickens using discontinuous sucrose gradients. Subsequently, the purified oocysts were suspended in 2.5% potassium dichromate solution at a concentration of 1 × 10⁷ organism/ml, and their viabilities were assessed by nucleic acid staining, histologic examination, and infectivity to 2-day-old chickens. All chickens inoculated with oocysts that had been stored for 1-18 months developed patent infections, while chickens infected with older oocysts remained uninfected. Between 5.8% and 82.2% of the oocysts, stored at 4℃ in 2.5% potassium dichromate solution, were found to be viable, as determined by nucleic acid staining. Parasite colonization in the bursa of Fabricius was detected in the microvillus border of bursal epithelium. The finding that C. baileyi oocysts remain infective to chickens for at least 18 months offers important time-saving advantages to investigators who frequently require large numbers of oocysts.

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Nicolas Valeix, Damien Costa, Louise Basmaciyan, Stéphane Valot, Anne Vincent, Romy Razakandrainibe, Florence Robert-Gangneux, Céline Nourrisson, Bruno Pereira, Emilie Fréalle, Philippe Poirier, Loic Favennec, Frederic Dalle
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A chicken embryo model for the maintenance and amplification of Cryptosporidium parvum and Cryptosporidium baileyi oocysts
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ELISA detection of vivax malaria with recombinant multiple stage-specific antigens and its application to survey of residents in endemic areas: Sera Kim, Hye-Jin Ahn, Tong-Soo Kim, Ho-Woo Nam; Korean J Parasitol 2003;41(4):203-207.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.203

Abstract
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An ELISA was developed for the diagnosis of vivax malaria using multiple stage-specific recombinant antigens of Plasmodium vivax. The DNA from the whole blood of a malaria patient was used as template to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1). Each amplified DNA fragment was inserted into pQE30 plasmid to induce the expression of His-tagged protein in Escherichia coli (M15 strain) by IPTG. His-tagged proteins were purified by Ni-NTA metal-affinity chromatography and used as antigens for ELISA with patient sera that were confirmed previously by blood smear examinations. When applied to patient sera, 122 (80.3%) out of 152 vivax malaria cases reacted to at least one antigen, while no reactions were observed with 128 uninfected serum samples. We applied this ELISA to the screening of 3,262 civilian residents in endemic regions near the DMZ, which resulted in 236 positively detected (7.2%) cases. This method can be applied to serological diagnosis and mass screening in endemic regions, or can be used as a safety test for transfusion blood in endemic areas.

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Evolutionary course of CsRn1 long-terminal-repeat retrotransposon and its heterogeneous integrations into the genome of the liver fluke, Clonorchis sinensis: Young-An Bae, Yoon Kong; Korean J Parasitol 2003;41(4):209-219.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.209

Abstract
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The evolutionary course of the CsRn1 long-terminal-repeat (LTR) retrotransposon was predicted by conducting a phylogenetic analysis with its paralog LTR sequences. Based on the clustering patterns in the phylogenetic tree, multiple CsRn1 copies could be grouped into four subsets, which were shown to have different integration times. Their differential sequence divergences and heterogeneous integration patterns strongly suggested that these subsets appeared sequentially in the genome of C. sinensis. Members of recently expanding subset showed the lowest level of divergence in their LTR and reverse transcriptase gene sequences. They were also shown to be highly polymorphic among individual genomes of the trematode. The CsRn1 element exhibited a preference for repetitive, agenic chromosomal regions in terms of selecting integration targets. Our results suggested that CsRn1 might induce a considerable degree of intergenomic variation and, thereby, have influenced the evolution of the C. sinensis genome.

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Seon-Hee Kim, Yoon Kong, Young-An Bae
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PwRn1, a novel Ty3/gypsy-like retrotransposon of Paragonimus westermani: molecular characters and its differentially preserved mobile potential according to host chromosomal polyploidy
Young-An Bae, Jong-Sook Ahn, Seon-Hee Kim, Mun-Gan Rhyu, Yoon Kong, Seung-Yull Cho
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Divergent long-terminal-repeat retrotransposon families in the genome of Paragonimus westermani: Young-An Bae, Yoon Kong; Korean J Parasitol 2003;41(4):221-231.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.221

Abstract
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To gain information on retrotransposons in the genome of Paragonimus westermani, PCR was carried out with degenerate primers, specific to protease and reverse transcriptase (rt) genes of long-terminal-repeat (LTR) retrotransposons. The PCR products were cloned and sequenced, after which 12 different retrotransposon-related sequences were isolated from the trematode genome. These showed various degrees of identity to the polyprotein of divergent retrotransposon families. A phylogenetic analysis demonstrated that these sequences could be classified into three different families of LTR retrotransposons, namely, Xena, Bel, and Gypsy families. Of these, two mRNA transcripts were detected by reverse transcriptase-PCR, showing that these two elements preserved their mobile activities. The genomic distributions of these two sequences were found to be highly repetitive. These results suggest that there are diverse retrotransposons including the ancient Xena family in the genome of P. westermani, which may have been involved in the evolution of the host genome.

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Case Reports

A serologically diagnosed human case of cutaneous larva migrans caused by Ancylostoma caninum: In-Ho Kwon, Hyung-Su Kim, Jong-Hee Lee, Min-Ho Choi, Jong-Yil Chai, Fukumi Nakamura-Uchiyama, Yukifumi Nawa, Kwang-Hyun Cho; Korean J Parasitol 2003;41(4):233-237.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.233

Abstract
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A 15-year-old boy, who had recently arrived back from a trip to Cambodia for a missionary camp, presented with several serpiginous thread-like skin lesions that began as small papules on the left upper extremities 2 weeks before his visit to Hospital. The skin lesions were pruritic and erythematous, and had migrated to the chest and abdomen. The histopathological findings showed only lymphocytic and eosinophilic infiltrations in the dermis of the biopsied skin lesion. The patient's serum reacted strongly to the Ancylostoma caninum antigen by an ELISA method. Therefore, he was diagnosed with cutaneous larva migrans by A. caninum. After the oral administration of albendazole and ivermectin, the skin lesions resolved without recurrence. This is the first reported case of a cutaneous larva migrans caused by Ancylostoma canimum diagnosed serologically using ELISA in Korea.

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Strongyloidiasis associated with amebiasis and giardiaisis in an immunocompetent boy presented with acute abdomen: Ener Cagr? Dinleyici, Nihal Dogan, Birsen Ucar, Huseyin Ilhan; Korean J Parasitol 2003;41(4):239-242.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.239

Abstract
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Strongyloides stercoralis (SS) is an intestinal nematode that is mainly endemic in tropical and subtropical regions and sporadic in temperate zones. SS infection frequently occurs in people who have hematologic malignancies, HIV infection and in individuals undergoing immunosuppressive therapy. In this study, we report a 12-year-old immunocompetent boy who was admitted to our hospital with acute abdomen. Laboratory evaluation showed strongyloidiasis, amebiasis and giardiasis. Clinical and laboratory findings immediately improved with albendazole therapy. Therefore, when diarrhea with signs of acute abdomen is observed, stool examinations should be done for enteroparasitosis. This approach will prevent misdiagnosis as acute abdomen. Complete clinical improvement is possible by medical therapy without surgical intervention.

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Brief Communication

Seroprevalence of antibodies to Neospora caninum in dogs and raccoon dogs in Korea: Jae-Hoon Kim, Min-Soo Kang, Byung-Chun Lee, Woo-Suk Hwang, Chang-Woo Lee, Byung-Jae So, J. P. Dubey, Dae-Yong Kim; Korean J Parasitol 2003;41(4):243-245.
Published online December 20, 2003; DOI: https://doi.org/10.3347/kjp.2003.41.4.243

Abstract
PDF

Neospora caninum is an important cause of abortion in cattle, and dogs are its only known definitive host. Its seroprevalence among domestic urban and rural dogs and feral raccoon dogs (Nyctereutes procyonoides koreensis) in Korea was studied by indirect fluorescent antibody test (IFAT) and by the neospora agglutination test (NAT), respectively. Antibodies to N. caninum were found in 8.3% of urban dogs and in 21.6% of dogs at dairy farms. Antibody titers ranged from 1:50 to 1:400. Antibodies to N. caninum were found in six (23%) of 26 raccoon dogs. However, the potential role of raccoon dogs as a source of horizontal transmission of bovine neosporosis needs further investigation. The results of this study suggest that there is a close relationship between N. caninum infection among dairy farm dogs and cattle in Korea. This study reports for the first time upon the seroprevalence of N. caninum infection in raccoon dogs in Korea.

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