• Journal of Microbiology
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• 제44권3호
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• pp.360-362
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• 2006

Cyclospora cayetanensis is an agent of emerging infectious disease, and a recognized cause of diarrhea in some patients. Also, the flagellated protozoan, Giardia intestinalis, induces a diarrheal illness of the small intestine. Cases of cyclosporiasis are frequently missed, primarily due to the fact that the parasite can be quite difficult to detect in human fecal samples, despite an increasing amount of data regarding this parasite. On the other hand, G. intestinalis can be readily recognized via the microscopic visualization of its trophozoite or cyst forms in stained preparations or unstained wet mounts. In this report, we describe an uncommon case of co-infection with G. intestinalis and C. cayetanensis in an immunocompetent patient with prolonged diarrhea, living in a non-tropical region of Turkey.

• 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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• 제58권5호
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• pp.589-592
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• 2020

Cyclospora cayetanensis is an apicomplexan protozoan and is one of the most common pathogens causing chronic diarrhea worldwide. Eight stool samples with diarrheal symptom out of 18 Korean residents who traveled to Nepal were obtained, and examined for 25 enteropathogens including 16 bacterial species, 5 viral species, and 4 protozoans in stool samples as causative agents of water-borne and food-borne disease. Only C. cayetanensis was detected by nested PCR, and 3 PCR-positive samples were sequenced to confirm species identification. However, the oocysts of C. cayetanensis in fecal samples could not be detected by direct microscopy of the stained sample. As far as we know, this is the first report of a group infection with C. cayetanensis from a traveler visiting Nepal, and the second report of a traveler's diarrhea by C. cayetanensis imported in Korea.

• Parasites, Hosts and Diseases
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• 제54권5호
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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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• 제56권5호
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• pp.419-427
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• 2018

This study aimed to develop a new multiplex real-time PCR detection method for 3 species of waterborne protozoan parasites (Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis) identified as major causes of traveler's diarrhea. Three target genes were specifically and simultaneously detected by the TaqMan probe method for multiple parasitic infection cases, including Cryptosporidium oocyst wall protein for C. parvum, glutamate dehydrogenase for G. lamblia, and internal transcribed spacer 1 for C. cayetanensis. Gene product 21 for bacteriophage T4 was used as an internal control DNA target for monitoring human stool DNA amplification. TaqMan probes were prepared using 4 fluorescent dyes, $FAM^{TM}$, $HEX^{TM}$, $Cy5^{TM}$, and CAL Fluor $Red^{(R)}$ 610 on C. parvum, G. lamblia, C. cayetanensis, and bacteriophage T4, respectively. We developed a novel primer-probe set for each parasite, a primer-probe cocktail (a mixture of primers and probes for the parasites and the internal control) for multiplex real-time PCR analysis, and a protocol for this detection method. Multiplex real-time PCR with the primer-probe cocktail successfully and specifically detected the target genes of C. parvum, G. lamblia, and C. cayetanensis in the mixed spiked human stool sample. The limit of detection for our assay was $2{\times}10$ copies for C. parvum and for C. cayetanensis, while it was $2{\times}10^3$ copies for G. lamblia. We propose that the multiplex real-time PCR detection method developed here is a useful method for simultaneously diagnosing the most common causative protozoa in traveler's diarrhea.

• Parasites, Hosts and Diseases
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• 제54권3호
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• pp.339-344
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• 2016

The genus Sarcocystis is not usually considered as an important enteric pathogen in immune compromised patients. It might be expected that species for which humans are the final host (Sarcocystis hominis and Sarcocystis suihominis as well as possibly others) would be encountered increasingly often in immunodeficient persons. This study aimed to address how to detect and differentiate Sarcocystis oocysts and/or sporocysts from enteric protozoans in the diarrheal samples of immunodeficient patients in Shiraz, Iran. Diarrheal samples of 741 immunodeficient patients with recurrent persistent or chronic diarrhea were examined by microscopy and molecular biological analysis. Oocysts-positive samples were 68 Cryptosporidium spp., 9 Cystoisospora belli (syn. Isospora belli), 2 Cyclospora cayetanensis, and 15 microsporidia (Enterocytozoon bieneusi). Sarcocystis-like sporocysts found from a woman were identified as Sarcocystis cruzi through 18S rDNA amplification and phylogenetic analysis. To the best of our knowledge, this is the first report of S. cruzi from a human.

• Parasites, Hosts and Diseases
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• 제56권4호
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• pp.371-374
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• 2018

A 3-month-old female Maltese puppy was hospitalized with persistent diarrhea in a local veterinary clinic. Blood chemistry and hematology profile were analyzed and fecal smear was examined. Diarrheal stools were examined in a diagnostic laboratory, using multiplex real-time polymerase chain reaction (PCR) against 23 diarrheal pathogens. Sequence analysis was performed using nested PCR amplicon of 18S ribosomal RNA. Coccidian oocysts were identified in the fecal smear. Although multiplex real-time PCR was positive for Cyclospora cayetanensis, the final diagnosis was Cystoisospora ohioensis infection, confirmed by phylogenetic analysis of 18S rRNA. To our knowledge, this the first case report of C. ohioensis in Korea, using microscopic examination and phylogenetic analysis.

• 한국동물위생학회지
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• 제45권2호
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• pp.101-110
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• 2022

This study was performed to investigate infectious gastrointestinal diseases in 115 Korean cats (83 indoors and 32 outdoors) with digestive signs such as diarrhea, anorexia or abdominal distention. Detection of infectious pathogens was analyzed using real-time PCR. As a result, 85 of 115 Korean cats were detected with feline corona virus (FCoV), feline parvo virus, Group A rotavirus, Clostridium perfringens (C. perfringens), Campylobacter coli (C. coli), Campylobacter jejuni, enterotoxigenic Escherichia coli, enteropathogenic Escherichia coli, Salmonella spp., Tritrichomonas foetus, Cyclospora cayetanensis, and Giardia lamblia. The most frequently detected pathogen was C. perfringens (52 cats, 61.2%), followed by FCoV (43 cats, 50.6%) and C. coli (16 cats, 18.8%). Also, single infection was the most common (43 cats), followed by double infection in 31 cats, triple infection in 7 cats, and quadruple infection in 4 cats. There was no significant relationship between pathogen detection and age, gender, living environment, weather, and diarrhea. However, there was a significant difference between the age group under 1 year and the age group 1~7 (P value<0.05). In this study, cats with suspected gastrointestinal infection were randomly evaluated, and other factors that could affect pathogen detection were insufficiently considered. For this reason, additional epidemiological investigations with a larger number of cats and sufficient consideration of the causes that may affect the results are needed. Nevertheless, it is thought that this study can also provide valuable information on gastrointestinal pathogens in Korean cats.

• 한국식품위생안전성학회지
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• 제37권2호
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• pp.87-96
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• 2022

본 연구에서는 온라인 및 오프라인 마켓에 유통 중인 100건의 신선편의 절단 과일을 계절별로 수집하고 50건의 신선편의 절단 채소와 50건의 밀 키트(월남 쌈 및 무 쌈) 내 절단 채소에 대하여 위생지표세균, 효모 및 곰팡이를 정량적으로 분석하였으며, 5개의 식중독균(B. cereus, S. aureus, L. monocytogenes, Salmonella spp., E. coli O157:H7)을 정성 및 정량적으로 분석하였다. 전체적으로 밀 키트 채소의 일반세균, 대장균군, 효모 및 곰팡이의 평균 오염수준이 신선편의 절단 채소 및 절단 컵 과일보다 높은 것으로 나타났다. 대장균은 두 식품 유형에서 모두 검출되지 않았으며 병원성 식중독균의 경우, S. aureus만 총 200건 중 6건이 검출되었는데(3%), 절단 과일 중 파인애플 1건에서 S. aureus가 0.6 log CFU/g로 검출이 되었고, 분리된 균주는 non-enterotoxin인 것으로 확인되었다. 신선편의 절단 채소 중 절단 채소 셀러리에서 1건, 밀 키트 채소 4건에서 S. aures 정성 검출되었으나, 정량적 분석 결과 밀 키트 채소인 적색 양배추 1건에서만 4.8 CFU/g 오염수준을 확인 하였으나 즉석섭취·편의식품류의 S. aureus 규격(100 CFU/g) 이하임을 확인하였다. 절단 채소에서 검출된 S. aureus의 독소형을 분석한 결과 셀러리와 적색 양배추에서 분리된 균이 각각 seg 와 sei 독소형임을 확인하였다. 따라서 절단 채소 제품 공정에서, 특히 밀 키트 채소에서의 황색포도상구균의 오염에 대하여 더 엄격한 관리방안이 필요한 것으로 확인되었다. 본 연구에서 분석한 50여종의 신선편의 식품과 밀 키트 제품에서는 모두 원포자충 및 작은와 포자충은 검출되지 않았으나 원충류에 의한 식중독 사고를 예방하기 위해 신선편의 식품의 세척 과정에 대한 지속적인 주의가 필요할 것으로 사료된다.