• 한국물환경학회지
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• 제24권6호
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• pp.817-822
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• 2008

In order to identify various Cryptosporidium species in environment, nested PCR-RFLP and DNA sequencing method were used. The sensitivity of nested PCR-RFLP based on 18s rRNA gene was shown to 1 oocyst. Therefore, we applied nested PCR-RFLP method to environmental samples. As a result, only 4 samples out of 8 samples confirmed as Cryptosporidium parvum by standard method of Cryptosporidium were identified as Cryptosporidium parvum by nested PCR-RFLP and DNA sequencing method. The rest of 4 samples among 8 samples were identified as Cryptosporidium muris, Cryptosporidium bailey. Therefore, in addition to standard method of Cryptosporidium, supplementary verification through nested PCR-RFLP and DNA sequencing should be needed to give more accurate information about risk of Cryptosporidium.

• 한국환경보건학회지
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• 제35권3호
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• pp.187-190
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• 2009

Cryptosporidium spp. were detected in 17 of 135 swine lagoon samples from five farms by 18S ribosomal DNA locus and PCR. Seventeen positive samples identified were included two distinctive genotypes C. suis and Cryptosporidium sp. based on a small-subunit rRNA gene-based PCR-restriction fragment length polymorphism analysis. Cryptosporidium spp. were detected out of farrowing, farrowing and nursery (mix), and finishing. Prevalence rate was 12.6% with infection rates between 3.7 and 18.5%. We concluded that Cryptosporidium oocysts can persist in treated lagoon and potentially contaminate surface water through improper discharge. This study was undertaken for the evaluation of the infection status of the genotypes of Cryptosporidium spp. in swine lagoon.

• Parasites, Hosts and Diseases
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• 제45권2호
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• pp.133-137
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• 2007

For evaluation of the prevalence of Cryptosporidium sp. infection in diarrheic and non-diarrheic humans in Iran, fecal specimens from diarrheic (n = 129) and non-diarrheic humans (n = 271) were collected and examined for the presence of Cryptosporidium sp. oocysts. The presence of Cryptosporidium sp. oocysts was determined by Ziehl-Neelsen acid-fast staining. Humans were grouped according to their age as follows: younger than 15, 16-25, 26-35, 36-50, and over 51 years. The results showed that the overall prevalence of infection in all 400 samples was 10.8%, but the prevalence (25.6%) in diarrheic humans was higher than that (3.7%) in non-diarrheic humans. Oocysts of Cryptosporidium sp. were detected in the feces of 21.4%, 9.3%, 8.8%, 6.7% and 5.7% of different age groups, respectively. The intensity of oocysts was significantly higher in diarrheic humans than in non-diarrheic ones. There was a significant association between Cryptosporidium sp. infection and occurrence of diarrhea (P < 0.05). The results indicate that Cryptosporidium sp. infection is prevalent in diarrheic humans in Iran.

• 대한환경공학회지
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• 제28권8호
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• pp.843-851
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• 2006

년도별 검출결과 2001년도에 각 지점별로 많은 검출을 보였지만 2004년도로 갈수록 Cryptosporidium과 Giardia 모두 10(oo)cysts/20 L 이하의 낮은 검출을 보였으며, 계절별로는 여름철과 늦여름($6{\sim}9$월)이 가장 많이 검출되었으며 지점별로는 고령교가 가장 높게 검출되었다. Cryptosporidium과 수질지표분석항목은 낮은 상관성을 보였으나 탁도와의 상관성에서는 Cryptosporidium의 경우 $R^2$=0.4914, Giardia는 $R^2$=0.6176으로 높게 나타났으며 Cryptosporidium보다 Giardia가 더 높은 상관성을 보였다. 그리고 탁도가 20 NTU 이상 일 때 Cryptosporidium과 Giardia의 검출이 대체로 20(oo)cystsd/20L 이상 높게 나타남을 알 수 있었다. Cryptosporidium과 Giardia의 상관성은 $R^2$=0.8432로 높게 나타났다. 각 지점별로 Cryptosporidium과 Giardia는 동시에 검출되고 있었으며, Giardia의 농도가 대체로 높게 나타났다. 축산폐수나 하수의 영향이 큰 것으로 나타났다.

• Parasites, Hosts and Diseases
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• 제54권4호
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• pp.423-429
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• 2016

Cryptosporidium is an important pathogen causing gastrointestinal disease in snakes and is distributed worldwide. The main objectives of this study were to detect and identify Cryptosporidium species in captive snakes from exotic pet shops and snake farms in Thailand. In total, 165 fecal samples were examined from 8 snake species, boa constrictor (Boa constrictor constrictor), corn snake (Elaphe guttata), ball python (Python regius), milk snake (Lampropeltis triangulum), king snake (Lampropeltis getula), rock python (Python sebae), rainbow boa (Epicrates cenchria), and carpet python (Morelia spilota). Cryptosporidium oocysts were examined using the dimethyl sulfoxide (DMSO)-modified acid-fast staining and a molecular method based on nested-PCR, PCR-RFLP analysis, and sequencing amplification of the SSU rRNA gene. DMSO-modified acid-fast staining revealed the presence of Cryptosporidium oocysts in 12 out of 165 (7.3%) samples, whereas PCR produced positive results in 40 (24.2%) samples. Molecular characterization indicated the presence of Cryptosporidium parvum (mouse genotype) as the most common species in 24 samples (60%) from 5 species of snake followed by Cryptosporidium serpentis in 9 samples (22.5%) from 2 species of snake and Cryptosporidium muris in 3 samples (7.5%) from P. regius.

• Parasites, Hosts and Diseases
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• 제56권3호
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• pp.281-285
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• 2018

Cryptosporidium is the most common protozoan that can infect a wide range of animals, including mammals and birds. Avian Cryptosporidium spp. can cause enteric and respiratory diseases which can be fatal in birds and some species are zoonotic. Companion birds have the potential as reservoir due to their close contact with humans. Pet shops are the major source of companion birds. However, few reports are available regarding Cryptosporidium spp. infection among companion birds kept in pet shops. The present study reports the prevalence and molecular characteristics of Cryptosporidium spp. among companion birds kept in pet shops in Japan. A total of 265 fresh fecal samples were obtained from birds kept in 4 pet shops; these birds belonged to 41 species in 3 bird orders. A nested polymerase chain reaction (PCR) assay targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. A total of 24 samples (9.1%) were positive, and Cryptosporidium spp. were detected from all pet shops. The prevalence of Cryptosporidium spp. in each of the bird orders was 6.5% (10/153) in Psittaciformes, 14.4% (13/90) in Passeriformes, and 4.5% (1/22) in Galliformes. Based on sequence analysis, 13 (54.2%) isolates were classified to C. galli, 8 (33.3%) were avian genotype III, and the remaining 3 (12.5%) were C. baileyi. No infection with zoonotic C. meleagridis and no coinfection with multiple Cryptosporidium spp. and/or genotypes were observed. The zoonotic potential of Cryptosporidium spp. infecting companion birds kept in pet shops in Japan is likely to be low.

• 한국임상수의학회지
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• 제24권4호
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• pp.588-592
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• 2007

The prevalence of E. coli(K99), Clostridium perfringens and Cryptosporidium parvum on acute diarrhea in suckling Korean native calves was evaluated in the field by a veterinary practice. In diagnosis, fecal samples were directly collected from calves that had diarrhea between 2 and 98 days of age. 40 samples were analyzed in October, 2006 and December, 2006. Clostridium perfringens and Cryptosporidium parvum were detected in 15(37.5%) and 4(10.0%) of the samples from diarrhetic calves, respectively. However, E. coli(K99) was not detected in the samples from diarrhetic calves. There was no significant difference(p>0.05) between October(5, 25.0%) and December(10, 50.0%) in incidence of detected Clostridium perfringens from diarrhetic calves. On the other hand, significant differences (p<0.05) in the detection rate of Clostridium perfringens were found between the within 1 month age and all other age groups. In the detection of Cryptosporidium parvum, there was no significant difference(p<0.05) between October (2, 10.0%) and December(2, 10.0%) in the incidence of detected Cryptosporidium parvum from diarrhetic calves. These results suggest that causative agents of calf diarrhea occurred frequently with Clostridium perfringens infection than E. coli(K99) and Cryptosporidium parvum.

• Parasites, Hosts and Diseases
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• 제48권2호
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• pp.105-111
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• 2010

To understand the situation of water contamination with Cryptosporidium spp. and Giardia spp. in the northern region of Portugal, we have established a long-term program aimed at pinpointing the sources of surface water and environmental contamination, working with the water-supply industry. Here, we describe the results obtained with raw water samples collected in rivers of the 5 hydrographical basins. A total of 283 samples were analyzed using the Method 1623 EPA, USA. Genetic characterization was performed by PCR and sequencing of genes 18S rRNA of Cryptosporidium spp. and $\beta$-giardin of Giardia spp. Infectious stages of the protozoa were detected in 72.8% (206 of 283) of the water samples, with 15.2% (43 of 283) positive for Giardia duodenalis cysts, 9.5% (27 of 283) positive for Cryptosporidium spp. oocysts, and 48.1% (136 of 283) samples positive for both parasites. The most common zoonotic species found were G. duodenalis assemblages A-I, A-II, B, and E genotypes, and Cryptosporidium parvum, Cryptosporidium andersoni, Cryptosporidium hominis, and Cryptosporidium muris. These results suggest that cryptosporidiosis and giardiasis are important public health issues in northern Portugal. To the authors' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in raw water samples in the northern region of Portugal.

• Parasites, Hosts and Diseases
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• 제55권2호
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• pp.137-142
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• 2017

Cryptosporidium and Cyclospora are well-known coccidian protozoa that can cause waterborne and food-borne 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.

• Parasites, Hosts and Diseases
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• 제57권2호
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• pp.197-200
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• 2019

Cryptosporidium is a common intestinal protozoan that can lead to diarrhea in humans and dogs. The predominant species of infection are C. hominis and C. parvum in humans, and C. canis in dogs. However, C. canis can infect immunocompromised humans. Considering the close contact with humans, dogs have the potential to be reservoirs for human cryptosporidiosis. Breeding kennels are the major supply source of puppies for pet shops. The present study is to determine the molecular prevalence and characteristics of Cryptosporidium spp. found in breeding kennel dogs. A total of 314 fecal samples were collected from young and adult dogs kept in 5 breeding kennels. A polymerase chain reaction targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. To determine the species, the DNA sequences were compared to GenBank data. Overall, 21.0% of the fecal samples were positive for Cryptosporidium spp. infection. Cryptosporidium spp. was detected in all 5 facilities. A sequencing analysis demonstrated that all isolates shared 99-100% similarity with C. canis. The results suggest that Cryptosporidium spp. infection is present at a high-level in breeding kennel dogs. However, because dominant species in this survey was C. canis, the importance of breeding kennel dogs as reservoirs for Cryptosporidium spp. transmission to humans is likely to be low in Japan.