This study has been conducted to estimate the occurrence of Cryptosporidium oocysts in water supplies in the Metropolitan area of Seoul, South Korea, for 10 years from 2000 to 2009. Water samples were collected quarterly at 6 intakes in the Han River and its largest stream and 6 conventional Water Treatment Plants (WTPs) serving drinking water for 10 million people of Seoul. Cryptosporidium oocysts were found in 22.5% of intake water samples and arithmetic mean was 0.65 oocysts/10 L (range 0-22 oocysts/10 L). Although the annual mean of oocyst number was as low as 0.04-1.90 oocysts/10 L, 3 peaks in 2004 and 2007 were observed and the pollution level was a little higher in winter. The lowest density was observed at Paldang intake and the pollution level increased at Kuui and Jayang intakes. At the end of the largest stream, oocysts were found in 70% of collected samples (mean 5.71 oocysts/10 L) and it seemed that its joining the Han River resulted in the increase at Kuui intake and downstream. Oocyst removal by physical process exceeded 2.0-2.3 log and then all finished water samples collected at 6 WTPs were negative for Cryptosporidium in each 100 L sample for 10 years. These results suggested that domestic wastewater from the urban region could be a source of Cryptosporidium pollution and separating sewage systems adjacent to the intakes could be meaningful for some intakes having weakness related to parasitological water quality.
Cryptosporidium/growth & development/*isolation & purification ; Fresh Water/*parasitology ; Oocysts/growth & development ; Republic of Korea ; Rivers/parasitology ; Water Supply/*analysis

In order to determine the distribution and prevalence of human cryptosporidiosis on western and southern coastal islands of Jeollanam-do (Province), fecal samples were collected from 2,541 people residing on 25 islands, 13 in the western coasts and 12 in the southern coasts, during July and August 2000. Fecal smears were prepared following formalin-ether sedimentation of the samples and stained by a modified acid-fast procedure. The presence of Cryptosporidium oocysts was determined by light microscopy. Cryptosporidium oocysts were detected in 38 specimens (1.5%). The oocyst positive rate varied (0-6.0%) according to island; the highest was detected on Oenarodo (6.0%), followed by Naenarodo (5.6%) and Nakwoldo (5.4%). The majority (35 persons, 94.6%) of Cryptosporidium-infected individuals were older than 50 years of age. Men (22/1,159; 1.9%) were infected at a higher rate than women (16/1,382; 1.2%). The results of the present survey indicate that human Cryptosporidium infections (due to Cryptosporidium hominis and/or C. parvum) are maintained at a relatively low prevalence on coastal islands of Jeollanam-do, Republic of Korea.
Sex Distribution ; Prevalence ; Oocysts ; Middle Aged ; Male ; Korea/epidemiology ; Infant, Newborn ; Infant ; Humans ; Female ; Feces/parasitology ; Cryptosporidium parvum/growth & development/*isolation & purification ; Cryptosporidium/growth & development/*isolation & purification ; Cryptosporidiosis/*epidemiology/parasitology ; Child, Preschool ; Child ; Animals ; Aged ; Age Distribution ; Adult ; Adolescent

Cryptosporidium, a coccidian parasite first described by Tyzzer (1907) from a laboratory mouse, has become an important human enteric pathogen causing overwhelming diarrhea especially in immunocompromised patients such as AIDS. This parasite has been reported from over 20 countries and is recognized as a cosmopolitan species. In Korea, however, there has been no report on human as well as animal cryptosporidiosis. This study was performed so as to verify the presence of Cryptosporidium in Korea by activating the parasite from laboratory mice by immunosuppression. Total 65 conventionally-bred ICR mice including a control (5 mice) and 3 experimental groups (20 each) were used for this study. Group I was immunosuppressed with prednisolone injection (1 mg IM, every other day) for 7 weeks. Group II (prednisolone injection and tetracycline administration) and Group III (prednisolone injection and trimethoprim-sulfamethoxazole administration) were prepared to observe the effect of antibacterial agents on the activation of cryptosporidiosis. In fecal examinations of mice Cryptosporidium oocysts (4-6 microns in size) were detected from 1 week after the start of immunosuppression and the mice began to die. In H-E stained tissue sections of the lower jejunum, numerous very small (2-4 microns), dense, ovoid or spherical, slightly basophilic bodies were seen attached on the free border of mucosal epithelial cells. In scanning and transmission electron microscopic observations, these organisms were identified as various developmental stages of Cryptosporidium. The species is considered to be C. parvum.(ABSTRACT TRUNCATED AT 250 WORDS)
Cryptosporidiosis-etiology ; Cryptosporidiosis-immunology ; Cryptosporidium-growth-and-development ; English-Abstract ; Immune-Tolerance ; Intestinal-Diseases,-Parasitic-etiology ; Intestinal-Diseases,-Parasitic-immunology ; Mice- ; Mice,-Inbred-ICR ; *Cryptosporidiosis-parasitology ; *Cryptosporidium-pathogenicity ; *Immunosuppression- ; *Intestinal-Diseases,-Parasitic-parasitology

From May to June 2012, a waterborne outbreak of 124 cases of cryptosporidiosis occurred in the plumbing systems of an older high-rise apartment complex in Seoul, Republic of Korea. The residents of this apartment complex had symptoms of watery diarrhea and vomiting. Tap water samples in the apartment complex and its adjacent buildings were collected and tested for 57 parameters under the Korean Drinking Water Standards and for additional 11 microbiological parameters. The microbiological parameters included total colony counts, Clostridium perfringens, Enterococcus, fecal streptococcus, Salmonella, Shigella, Pseudomonas aeruginosa, Cryptosporidium oocysts, Giardia cysts, total culturable viruses, and Norovirus. While the tap water samples of the adjacent buildings complied with the Korean Drinking Water Standards for all parameters, fecal bacteria and Cryptosporidium oocysts were detected in the tap water samples of the outbreak apartment complex. It turned out that the agent of the disease was Cryptosporidium parvum. The drinking water was polluted with sewage from a septic tank in the apartment complex. To remove C. parvum oocysts, we conducted physical processes of cleaning the water storage tanks, flushing the indoor pipes, and replacing old pipes with new ones. Finally we restored the clean drinking water to the apartment complex after identification of no oocysts.
Cryptosporidiosis/*epidemiology/*parasitology ; Cryptosporidium parvum/genetics/growth & development/*isolation & purification ; Disease Outbreaks ; Drinking Water/*parasitology ; Housing ; Humans ; Oocysts/growth & development ; Republic of Korea/epidemiology ; Water Supply/analysis

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.
Animals ; Cryptosporidiosis/parasitology ; Cryptosporidium parvum/*growth & development/radiation effects/ultrastructure ; Female ; Gamma Rays ; Humans ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Transmission ; Oocysts/growth & development/*radiation effects/*ultrastructure

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.
Adenocarcinoma ; Animals ; Cell Line, Tumor ; Cryptosporidium/*growth & development ; Culture Media ; Human ; Hydrogen-Ion Concentration ; Stomach/*parasitology ; Stomach Neoplasms ; Support, Non-U.S. Gov't

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 degrees C 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 x 10 (7) 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 degrees C 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.
Animals ; Bursa of Fabricius/parasitology ; Chickens/*parasitology ; Coloring Agents ; Cryptosporidiosis/parasitology/pathology/*veterinary ; Cryptosporidium/drug effects/*growth & development/pathogenicity ; Feces/parasitology ; Oocysts/drug effects/*growth & development/pathogenicity ; *Organic Chemicals ; *Potassium Dichromate/pharmacology ; Poultry Diseases/parasitology/pathology ; Preservation, Biological/*methods ; Staining and Labeling

Despite the development of new technologies, new challenges still remain for large scale proteomic profiling when dealing with complex biological mixtures. Fractionation prior to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis is usually the preferred method to reduce the complexity of any biological sample. In this study, a gel LC-MS/MS approach was used to explore the stage specific proteome of Cryptosporidium (C.) parvum. To accomplish this, the sporozoite protein of C. parvum was first fractionated using SDS-PAGE with subsequent LC-MS/MS analysis. A total of 135 protein hits were recorded from 20 gel slices (from same gel lane), with many hits occurring in more than one band. Excluding all non-Cryptosporidium entries and proteins with multiple hits, 33 separate C. parvum entries were identified during the study. The overall goal of this study was to reduce sample complexity by protein fractionation and increase the possibility of detecting proteins present in lower abundance in a complex protein mixture.
Chemical Fractionation/methods ; Chromatography, Liquid/methods/veterinary ; Cryptosporidium parvum/*chemistry/growth & development/metabolism ; Electrophoresis, Polyacrylamide Gel/methods/veterinary ; Gene Expression Profiling/*methods/veterinary ; Proteome/analysis ; Proteomics/*methods ; Protozoan Proteins/*analysis ; Sporozoites/chemistry/metabolism ; Tandem Mass Spectrometry/methods/veterinary