Parasites, Hosts and Diseases

"Woo-Yoon Park"

Original Article

Simultaneous Molecular Detection of Cryptosporidium and Cyclospora from Raw Vegetables in Korea: Seobo Sim, Jua Won, Jae-Whan Kim, Kyungjin Kim, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2017;55(2):137-142.
Published online April 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.2.137

Abstract
PDF

Cryptosporidium and Cyclospora are well-known coccidian protozoa that can cause waterborne and foodborne diarrheal illnesses. There have been a few reports regarding contamination in different vegetables with Cryptosporidium, but no data are available regarding the sources of Cyclospora infections in Korea. In the present study, we collected 6 kinds of vegetables (perilla leaves, winter-grown cabbages, chives, sprouts, blueberries, and cherry tomatoes) from July 2014 to June 2015, and investigated contamination by these 2 protozoa using multiplex quantitative real-time PCR. Among 404 vegetables, Cryptosporidium and Cyclospora were detected in 31 (7.7%) and 5 (1.2%) samples, respectively. In addition, Cryptosporidium was isolated from all 6 kinds of vegetables, whereas Cyclospora was detected in 4 kinds of vegetables (except perilla leaves and chives). Cryptosporidium (17.8%) and Cyclospora (2.9%) had the highest detection rates in chives and winter-grown cabbages, respectively. Cryptosporidium was detected all year long; however, Cyclospora was detected only from October to January. In 2 samples (sprout and blueberry), both Cryptosporidium and Cyclospora were detected. Further investigations using TaqI restriction enzyme fragmentation and nested PCR confirmed Cryptosporidium parvum and Cyclospora cayetanensis, respectively. In conclusion, we detected C. cayetanensis in vegetables for the first time in Korea. This suggests that screening should be employed to prevent these protozoal infections in Korea.

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Brief Communication

Cryptosporidium hominis Infection Diagnosed by Real-Time PCR-RFLP: Hyeng-Il Cheun, Kyungjin Kim, Sejoung Yoon, Won-Ja Lee, Woo-Yoon Park, Seobo Sim, Jae-Ran Yu; Korean J Parasitol 2013;51(3):353-355.
Published online June 30, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.3.353

Abstract
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There are approximately 20 known species of the genus Cryptosporidium, and among these, 8 infect immunocompetent or immunocompromised humans. C. hominis and C. parvum most commonly infect humans. Differentiating between them is important for evaluating potential sources of infection. We report here the development of a simple and accurate real-time PCR-based restriction fragment length polymorphism (RFLP) method to distinguish between C. parvum and C. hominis. Using the CP2 gene as the target, we found that both Cryptosporidium species yielded 224 bp products. In the subsequent RFLP method using TaqI, 2 bands (99 and 125 bp) specific to C. hominis were detected. Using this method, we detected C. hominis infection in 1 of 21 patients with diarrhea, suggesting that this method could facilitate the detection of C. hominis infections.

Citations

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Iranian Red Crescent Medical Journal.2017;[Epub] CrossRef
Detection of Cryptosporidium parvum in Environmental Soil and Vegetables
Semie Hong, Kyungjin Kim, Sejoung Yoon, Woo-Yoon Park, Seobo Sim, Jae-Ran Yu
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Original Articles

Ultrastructural Changes in Cryptosporidium parvum Oocysts by Gamma Irradiation: Migyo Joung, Sejeong Yun, Mijoung Joung, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2011;49(1):25-31.
Published online March 18, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.1.25

Abstract
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Cryptosporidium parvum is known as one of the most highly resistant parasites to gamma irradiation. To morphologically have an insight on the radioresistance of this parasite, ultrastructural changes in C. parvum sporozoites were observed after gamma irradiation using various doses (1, 5, 10, and 25 kGy) following a range of post-irradiation incubation times (10 kGy for 6, 12, 24, 48, 72, and 96 hr). The ultrastructures of C. parvum oocysts changed remarkably after a 10-kGy irradiation. Nuclear membrane changes and degranulation of dense granules were observed with high doses over 10 kGy, and morphological changes in micronemes and rhoptries were observed with very high doses over 25 kGy. Oocyst walls were not affected by irradiation, whereas the internal structures of sporozoites degenerated completely 96 hr post-irradiation using a dose of 10 kGy. From this study, morphological evidence of radioresistance of C. parvum has been supplemented.

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Sandrine Boivin, Hiroshi Nagaoka, Takahiro Fujioka
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Multiplex PCR Detection of Waterborne Intestinal Protozoa: Microsporidia, Cyclospora, and Cryptosporidium: Seung-Hyun Lee, Migyo Joung, Sejoung Yoon, Kyoungjin Choi, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2010;48(4):297-301.
Published online December 16, 2010; DOI: https://doi.org/10.3347/kjp.2010.48.4.297

Abstract
PDF

Recently, emerging waterborne protozoa, such as microsporidia, Cyclospora, and Cryptosporidium, have become a challenge to human health worldwide. Rapid, simple, and economical detection methods for these major waterborne protozoa in environmental and clinical samples are necessary to control infection and improve public health. In the present study, we developed a multiplex PCR test that is able to detect all these 3 major waterborne protozoa at the same time. Detection limits of the multiplex PCR method ranged from 10¹ to 10² oocysts or spores. The primers for microsporidia or Cryptosporidium used in this study can detect both Enterocytozoon bieneusi and Encephalitozoon intestinalis, or both Cryptosporidium hominis and Cryptosporidium parvum, respectively. Restriction enzyme digestion of PCR products with BsaBI or BsiEI makes it possible to distinguish the 2 species of microsporidia or Cryptosporidium, respectively. This simple, rapid, and cost-effective multiplex PCR method will be useful for detecting outbreaks or sporadic cases of waterborne protozoa infections.

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Brief Communication

Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum: Jae-Ran Yu, Soo-Ung Lee, Woo-Yoon Park; Korean J Parasitol 2009;47(3):293-297.
Published online August 28, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.3.293

Abstract
PDF

Improved methods for detection of Cryptosporidium oocysts in environmental and clinical samples are urgently needed to improve detection of cryptosporidiosis. We compared the sensitivity of 7 PCR primer sets for detection of Cryptosporidium parvum. Each target gene was amplified by PCR or nested PCR with serially diluted DNA extracted from purified C. parvum oocysts. The target genes included Cryptosporidium oocyst wall protein (COWP), small subunit ribosomal RNA (SSU rRNA), and random amplified polymorphic DNA. The detection limit of the PCR method ranged from 10³ to 10⁴ oocysts, and the nested PCR method was able to detect 10⁰ to 10² oocysts. A second-round amplification of target genes showed that the nested primer set specific for the COWP gene proved to be the most sensitive one compared to the other primer sets tested in this study and would therefore be useful for the detection of C. parvum.

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Original Article

A Hospital-Based Serological Survey of Cryptosporidiosis in the Republic of Korea: Jong-Kyu Lee, Eun-Taek Han, Sun Huh, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2009;47(3):219-225.
Published online August 28, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.3.219

Abstract
PDF

The seroprevalence of cryptosporidiosis was examined using patients' sera collected from hospitals located in 4 different areas of the Republic of Korea. ELISA was used to measure antibody titers against Cryptosporidium parvum antigens from a total of 2,394 serum samples, which were collected randomly from patients in local hospitals; 1) Chungbuk National University Hospital, 2) Konkuk University Hospital, 3) local hospitals in Chuncheon, Gangwon-do (province), 4) Jeonnam National University Hospital, from 2002 through 2003. Of the 2,394 samples assayed, 34%, 26%, and 56% were positive for C. parvum-specific IgG, IgM, and IgA antibodies, respectively. Positive IgG titers were most common in sera from Jeonnam National University Hospital, Gwangju, Jeollanam-do, and positive IgM titers were most common in sera from Chungbuk National University Hospital, Cheongju, Chuncheongbuk-do. The seropositivity was positively correlated with age for both the IgG and IgA antibodies but was negatively correlated with age for the IgM antibodies. Western blotting revealed that 92%, 83%, and 77% of sera positive for IgG, IgM, and IgA ELISA reacted with 27-kDa antigens, respectively. These results suggested that infection with Cryptosporidium in hospital patients occurs more commonly than previously reported in the Republic of Korea.

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A Waterborne Outbreak and Detection of Cryptosporidium Oocysts in Drinking Water of an Older High-Rise Apartment Complex in Seoul
Eun-Joo Cho, Jin-Young Yang, Eun-Sook Lee, Se-Chul Kim, So-Yang Cha, Sung-Tek Kim, Man-Ho Lee, Sun-Hee Han, Young-Sang Park
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Brief Communication

Ultrastructural Localization of Cryptosporidium parvum Antigen Using Human Patients Sera: Jong-Gyu Lee, Eun-Taek Han, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2009;47(2):171-174.
Published online May 27, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.2.171

Abstract
PDF

The antigen location of Cryptosporidium parvum, which stimulates antibody formation in humans and animals, was investigated using infected human sera. Immuno-electron microscopy revealed that antigenicity-inducing humoral immunity was located at various developmental stages of parasites, including asexual, sexual stages, and oocysts. The amount of antigen-stimulating IgG antibodies was particularly high on the oocyst wall. The sporozoite surface was shown to give stimulation on IgG and IgM antibody formation. Trophozoites implicated the lowest antigenicity to humoral immunity, both IgG and IgM, by showing the least amount of gold labeling. Immunogold labeling also provided clues that antigens were presented to the host-cell cytoplasm via feeder organelles and host-parasite junctions.

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Cryptostatin, a chagasin-family cysteine protease inhibitor ofCryptosporidium parvum
J.-M. KANG, H.-L. JU, J.-R. YU, W.-M. SOHN, B.-K. NA
Parasitology.2012; 139(8): 1029. CrossRef

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Original Articles

Quantitative Evaluation of Infectivity Change of Cryptosporidium parvum after Gamma Irradiation: Soo-Ung Lee, Mikyo Joung, Taekyoung Nam, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2009;47(1):7-11.
Published online March 12, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.1.7

Abstract
PDF

Cryptosporidium parvum is a well-known waterborne and opportunistic intracellular protozoan parasite that causes diarrheal illness. In this study, we quantitatively investigated reduction of the infectivity of C. parvum after gamma irradiation and repair of the infectivity during incubation time after irradiation. C. parvum oocysts were subjected to gamma irradiation at various doses (1, 5, 10, and 25 kGy), and the in vitro infectivity was measured by real-time PCR every day up to 7 days after irradiation. The in vitro infectivity of C. parvum on human ileocecal adenocarcinoma cells (HCT-8) was effectively reduced (> 2 log₁₀) by irradiation at 10 kGy or more. However, in the experiment to find out repair of the infectivity, recovery was not noted until day 7 post-incubation.

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Ultrastructural Changes inCryptosporidium parvumOocysts by Gamma Irradiation
Migyo Joung, Sejeong Yun, Mijoung Joung, Woo-Yoon Park, Jae-Ran Yu
The Korean Journal of Parasitology.2011; 49(1): 25. CrossRef
Effect of Infrared Irradiation of Oocysts on the Pathology of Experimental Cryptosporidiosis in Apparently Normal and Immunosuppressed Mice
Hafidh I. Al-Sadi, Saevan S. Al-Mahm
Research Journal of Biological Sciences.2010; 5(2): 177. CrossRef
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Rejoining of gamma-ray-induced DNA damage in Cryptosporidium parvum measured by the comet assay
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Experimental Parasitology.2010; 125(3): 230. CrossRef

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67 Download
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In vitro culture of Cryptosporidium muris in a human stomach adenocarcinoma cell line: Min-Ho Choi, Sung-Tae Hong, Jong-Yil Chai, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2004;42(1):27-34.
Published online March 20, 2004; DOI: https://doi.org/10.3347/kjp.2004.42.1.27

Abstract
PDF

We investigated the optimal culture conditions for Cryptosporidium muris in a human stomach adenocarcinoma (AGS) cell line by determining the effects of medium pH and of selected supplements on the development of C. muris. The optimum pH of the culture medium required for the development of C. muris was determined to be 6.6. The number of parasites significantly increased during cultivation for 72 hr (p < 0.05) at this level. On the other hand, numbers decreased linearly after 24 hr of incubation at pH 7.5. When cultured in different concentrations of serum, C. muris in media containing 5% FBS induced 4-7 times more parasites than in 1% or 10% serum. Of the six medium supplements examined, only 1 mM pyruvate enhanced the number of C. muris in vitro. Transmission electron microscopic observation showed the developmental stages of C. muris in the cytoplasm of the cells, not in an extracytoplasmic location. The growth of C. muris in AGS cells provides a means of investigating its biological characteristics and of testing its response to therapeutic agents. However, a more optimized culture system is needed for the recovery of oocysts on a large scale in vitro.

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