• Parasites, Hosts and Diseases
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• v.53 no.6
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• pp.745-747
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• 2015

Cryptosporidium, a protozoan parasite that causes watery diarrhea, is found worldwide and is common in areas with low water hygiene. In February 2014, 866 stool samples were collected from the inhabitants of 2 rural areas in White Nile State, Sudan. These stool samples were assessed by performing modified acid-fast staining, followed by examination under a light microscope. The overall positive rate of Cryptosporidium oocysts was 13.3%. Cryptosporidium oocysts were detected in 8.6% stool samples obtained from inhabitants living in the area having water purification systems and in 14.6% stool samples obtained from inhabitants living in the area not having water purification systems. No significant difference was observed in the prevalence of Cryptosporidium infection between men and women (14.7% and 14.1%, respectively). The positive rate of oocysts by age was the highest among inhabitants in their 60s (40.0%). These findings suggest that the use of water purification systems is important for preventing Cryptosporidium infection among inhabitants of these rural areas in Sudan.

• Journal of Environmental Health Sciences
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• v.39 no.1
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• pp.32-42
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• 2013

Objectives: Cryptosporidium, a protozoan parasite, has been recognized as a frequent cause of waterborne disease due to its extremely strong resistance against chlorine disinfection. Although there has as yet been no report of a Cryptosporidium outbreak through drinking water in Korea, it is important to estimate the health risk of Cryptosporidium in water supply systems because of the various infection cases in human and domestic animals and frequent detection reports on their oocysts in water environments. Methods: This study evaluated the annual infection risk of Cryptosporidium in tap water using the quantitative microbial risk assessment technique. Exposure assessment was performed upon the results of a national survey on Cryptosporidium on the water sources of 97 large-scale water purification plants in Korea, water treatment efficacy, and daily unboiled tap water consumption. The estimates of the US Environmental Protection Agency on the mean likelihood of infection from ingesting one oocyst were applied for effect assessment. Results: Using probabilistic methods, mean annual infection risk of Cryptosporidiosis by the intake of tap water was estimated to fall within the range of $2.3{\times}10^{-4}$ to $1.0{\times}10^{-3}$ (median $5.7{\times}10^{-4}$). The risk in using river sources was predicted to be four times higher than with lake sources. With 0.5-log higher removal efficacy, the risk was estimated to be $1.8{\times}10^{-4}$, and could then be lowered by one-third. Conclusions: These estimations can be compared with acceptable risk and then used to determine the adequacy and priority of various drinking water quality strategies such as the establishment of new treatment technology.

• Parasites, Hosts and Diseases
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• v.53 no.2
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• pp.147-154
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• 2015

Various constituents in clinical specimens, particularly feces, can inhibit the PCR assay and lead to false-negative results. To ensure that negative results of a diagnostic PCR assay are true, it should be properly monitored by an inhibition control. In this study, a cloning vector harboring a modified target DNA sequence (${\approx}375bp$) was constructed to be used as a competitive internal amplification control (IAC) for a conventional PCR assay that detects ${\approx}550bp$ of the Cryptosporidium oocyst wall protein (COWP) gene sequence in human feces. Modification of the native PCR target was carried out using a new approach comprising inverse PCR and restriction digestion techniques. IAC was included in the assay, with the estimated optimum concentration of 1 fg per reaction, as duplex PCR. When applied on fecal samples spiked with variable oocysts counts, ${\approx}2$ oocysts were theoretically enough for detection. When applied on 25 Cryptosporidium-positive fecal samples of various infection intensities, both targets were clearly detected with minimal competition noticed in 2-3 samples. Importantly, both the analytical and the diagnostic sensitivities of the PCR assay were not altered with integration of IAC into the reactions. When tried on 180 randomly collected fecal samples, 159 were Cryptosporidium-negatives. Although the native target DNA was absent, the IAC amplicon was obviously detected on gel of all the Cryptosporidium-negative samples. These results imply that running of the diagnostic PCR, inspired with the previously developed DNA extraction protocol and the constructed IAC, represents a useful tool for Cryptosporidium detection in human feces.

• Korean Journal of Veterinary Research
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• v.51 no.2
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• pp.101-105
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• 2011

We investigate the resistance of Cryptosporidium (C.) parvum oocysts to commercial bleach treatment. The viability and infectivity of C. parvum oocysts suspended in 100, 50, 25, 12.5, 6.3 or 3.2% aqueous commercial bleach for 10, 30, 60, 120 or 180 min at room temperature were assessed by nucleic acid Syto-9 staining, histologic examination of ileum and infectivity to immunosuppressed neonatal C57BL/6N mice. Although the viability was decreased compared with normal oocysts, all oocysts in contact with serially diluted commercial bleach for 180 min were alive by nucleic acid dye Syto-9 staining. And, microscopic examination of ileum sections revealed developmental stages of C. parvum in all mice. The oocyst shedding patterns between mice infected with oocysts contacted with commercial bleach and normal control mice were not significantly different each other. Although commercial bleach is widely used as a bacterial and viral disinfectant, the present findings indicate that it is not an effective disinfectant for C. parvum oocysts under practical conditions. Authors conclude that, therefore, it is undesirable to recommend commercial bleach as a disinfectant for C. parvum oocysts.

• Parasites, Hosts and Diseases
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• v.51 no.4
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• pp.461-466
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• 2013

From May to June 2012, a waterborne outbreak of 124 cases of cryptosporidiosis occurred in the plumbing system 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 virus, 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.

• Parasites, Hosts and Diseases
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• v.53 no.6
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• pp.737-743
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• 2015

In order to examine the prevalence of Cryptosporidium infection in wild rodents and insectivores of South Korea and to assess their potential role as a source of human cryptosporidiosis, a total of 199 wild rodents and insectivore specimens were collected from 10 regions of South Korea and screened for Cryptosporidium infection over a period of 2 years (2012-2013). A nested-PCR amplification of Cryptosporidium oocyst wall protein (COWP) gene fragment revealed an overall prevalence of 34.2% (68/199). The sequence analysis of 18S rRNA gene locus of Cryptosporidium was performed from the fecal and cecum samples that tested positive by COWP amplification PCR. As a result, we identified 4 species/genotypes; chipmunk genotype I, cervine genotype I, C. muris, and a new genotype which is closely related to the bear genotype. The new genotype isolated from 12 Apodemus agrarius and 2 Apodemus chejuensis was not previously identified as known species or genotype, and therefore, it is supposed to be a novel genotype. In addition, the host spectrum of Cryptosporidium was extended to A. agrarius and Crosidura lasiura, which had not been reported before. In this study, we found that the Korean wild rodents and insectivores were infected with various Cryptosporidium spp. with large intra-genotypic variationa, indicating that they may function as potential reservoirs transmitting zoonotic Cryptosporidium to livestock and humans.

• Parasites, Hosts and Diseases
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• v.34 no.4
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• pp.255-258
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• 1996

Eight 2-day-old SPF Chickens were each inoculated Orally With 3 Sing1e dose Of 5 × 105 oocysts of Cryptosporinium boilevi. and immunoglobulin G (IgG) antibody responses were chronologically measured by indirect immunofluorescent antibody (IFA) assay. Anti-C. bcileyi IgG antibody levels remained high (1 : 106.67 to 1:512.00) for at least 4 months with 330 days of a detectable period. Ten days after the negative conversion, each chicken was re-challenged with 1 × 107 oocysts of the same species. Subsequent infection in 340-day-old individuals caused sudden elevated IgG antibody levels and the titer peaked on day 28 postchallenge inoculation (PCI), at 1:1.024 with a 65 days of detection period. Chickens in primary infection showed oocyst shedding profiles. but did not exhibit any oocyst shedding before or after experimental reinfection.

• Parasites, Hosts and Diseases
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• v.31 no.3
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• pp.223-230
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• 1993

Monoclonal antibodies (Ub) against merozoites and sporozoites of the protozoan parasite Cryptosporidium pcnjum were examined for potential modulation of cryptosporidial infections in vivo by daily oral Ub administration to oocyst-inoculated neonatal mice. Monoclonal-treated neonatal mice were sacrified four and eight days post infection (pi) . Differences in infection rates were observed among the treatment groups at the p < 0.05 level. Suckling mice treated daily with orally administered mixtures of mAbs( ascitic fluids) showed significantly reduced parasite loads compared to control mice at flour and eight days pi, while suckling mice receiving mAb Cmg-3 alone showed signiacant differences only at 4 days Pi., suggesting that passive transfer of mAb may be of value in controlling cryptosporidial infections.

• Parasites, Hosts and Diseases
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• v.54 no.5
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• pp.631-636
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• 2016

This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the major causes of traveler's diarrhea and are resistant to standard antimicrobial treatments. The target genes included the Cryptosporidium oocyst wall protein for C. parvum, Glutamate dehydrogenase for G. lamblia, and 18S ribosomal RNA (18S rRNA) for C. cayetanensis. The sizes of the amplified fragments were 555, 188, and 400 bps, respectively. The multiplex-touchdown PCR protocol using a primer mixture simultaneously detected protozoa in human stools, and the amplified gene was detected in > $1{\times}10^3$ oocysts for C. parvum, > $1{\times}10^4$ cysts for G. lamblia, and > 1 copy of the 18S rRNA gene for C. cayetanensis. Taken together, our protocol convincingly demonstrated the ability to simultaneously detect C. parvum, G. lamblia, and C. cayetanenesis in stool samples.

• Parasites, Hosts and Diseases
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• v.29 no.2
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• pp.149-160
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• 1991

Each of SPF mice(Scl: ICR strain, 3-week-old males) was inoculated with 5$\times$104 oocysts of Cryptosporidium by stomach tube. The oocysts were large type one which was previously isolated from Korean mice, and passaged in 3-week-old SPF mice. The patterns of oocyst discharge were monitored daily, and in order to observe the ultrastructure of developmental stages the stomach of the mice was examined by transmission electron microscopy (TEM) at 4 weeks post-inoculation. The prepatent period for 6 mice was 5.6 days post-inoculation on the average, and the patent period was 63.2 days. The number of oocysts discharged per day from the mice reached peak on day 36.6 post-inoculation on the average. A large number of oocysts were found in fecal samples obtained from inoculated mice on days 30~50 post-inoculation. C. tsuris was larger than C. parvum at almost every developmental stages, the sixte difference being 1.4 times in oocysts, 2.4 times in sporozoites, 1.6 times in merozoites, and 1.5 times in microgametes. The ultrastructural features of the attachment site of C. tsuris to the mucus cells were remarkably different from those of C. parvum and its closely related species. The anterior projection of the protozoa (C. muris), the outer aspect of which was surrounded by a thick filamentous process of the host cell, has not been reported at any developmental stages of C. parvum or its closely related species. The size of the oocysts of strain RN 66 was larger than that of Korean mice origin. The above results reveal that the large type Cryptosporidium of Korean mice origin is identified as Cryptosporidium muris and this type was named as C. muris (strain MCR).