• Parasites, Hosts and Diseases
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• v.58 no.5
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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.

• Journal of Environmental Health Sciences
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• v.48 no.3
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• pp.176-182
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• 2022

Background: The risk of imported infectious diseases has been increasing with the annual rise in the number of international travelers. Objectives: This study aims to analyze the distribution and characteristics of intestinal bacteria isolated in 2019 from residents of Chungcheongnam-do Province with experience of travelling overseas. Methods: Twenty-three former overseas travelers with diarrhea were analyzed to detect viruses and bacteria according to the Manual for Detection of Foodborne Pathogens at Outbreaks. Additionally, antibiotic susceptibility tests and 16s rRNA sequencing were performed. Results: Twenty-five strains of ten pathogens were isolated from 18 samples. Pathogenic E. coli was the most common at 57.7%, followed by Clostridium perfringens (15.4%), Campylobacter spp. (7.7%), and Salmonella spp. (7.7%). The serotype of Salmonella was confirmed as Salmonella Braenderup, II 9,46:g,[m],[s],t:[e,n,x]. Conclusions: It was confirmed that the major enteric bacterial pathogens isolated from overseas travelers in Chungcheongnam-do Province were pathogenic E. coli, as found in other studies. The study on Plesiomonas shigelloides is meaningful in that it is reported as a rare case of infection in Korea. Antibiotic resistance and 16s rRNA analysis were performed, which is expected to provide important basic data for the prevention of traveler's diarrhea.

• Parasites, Hosts and Diseases
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• v.56 no.5
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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.

• Journal of Dairy Science and Biotechnology
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• v.30 no.2
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• pp.139-143
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• 2012

For human including newborn baby, the intestinal microbiota can play an important role in the development of the intestinal mucosa and in maintaining the balance of the immune cells. Important functions of the intestinal microbiota include the inhibition of the colonization of the intestine by potentially pathogenic microorganisms. Thus, the research of probiotics have been focused on the prevention and treatment of disorders associated with the gastrointestinal tract (GIT), including pathogen infection, traveler's diarrhea, antibiotic-associated diarrhea, and constipation. Probiotics have also been suggested as therapeutic agents against irritable bowel syndrome and inflammatory bowel diseases. An increasing amount of evidence from clinical studies suggests that they are effective in the prevention of atopic allergies and may have potential anti-carcinogenic effects. Until recent years many scientific research for this use has been based on empirical observations. Therefore, probiotics in the form of fermented milk products have been long part of attempts to maintain good health in world wide.

• 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.

• The Journal of the Korean Society for Microbiology
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• v.18 no.1
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• pp.53-58
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• 1983

Enterotoxigenic E. coli is one of causative agents of the infantile diarrhea and traveler's diarrhea. A modified infant mouse assay(IMA) was developed for the detection of heat stable enterotoxin (ST) of E. coli isolated from diarrheal and control infants and assay system was established with using enterotoxin producing reference strains. The supernatant of the 24 hour-shaking culture of E. coli in Casamino Acid Yeast Extract Salt Broth(CYES-2) was ingested orally into the 2-4 day old ICR mice. After the mice were kept at $25^{\circ}C$ for 4 hours, they were sacrificed and the gut weight body weight ratio(GW/BW) was taken as the index of fluid accumulation induced by heat stable enterotoxin of E. coli. The results obtained were as follows; 1. The GW/BW responses of IMA tested with enterotoxin reference strains of E. coli(E. coli O148H28:$ST^+LT^+$, E. coli $O78H^-:ST^+LT^+$, E. coli O15H11:$ST^-LT^+$, E. coli O1H7:$ST^-LT^-$) appeared ta be ST dose-dependent, and not LT-dependent. From the dose-response curve, $25{\mu}l$ of culture supernatant was determined as test amount of the IMA. 2. Frequency distribution of IMA result from 643 strain of E. coli showed normal distribution at low GW/BW ratio and dispersed pattern at high GW/BW ratio. The GW/BW ratios of $0.056{\pm}0.004(mean{\pm}SD)$ of normal distribution which distributed from 0.044 to 0.068(P<0.01) was considered as ST negative. Thus the GW/BW ratio above 0.069 could be regarded as ST positive.

• The Journal of the Korean Society for Microbiology
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• v.17 no.1
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• pp.35-41
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• 1982

Enterotoxigenk E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-labile enterotoxin is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a marker for identification of the enterotoxigenic E. coli from non pathogenic E. coli. Therefore knowledge about the heat-labile enterotoxin is essential not only for understanding the pathogenesis but also for the diagnosis of the diarrhea. However the in-vitro heat-labile enterotoxin production is reported to be greatly affected by the cultural condition. In this regards, this study was designed to know the optimal conditions for the production of the heat-labile enterotoxin by assaying the permeability factor in the 18 hours culture supernatant of E. coli 08K25(B2) H9 and of E. coli 015 H11. Results obtained were summerized as follows: 1. Amounts of heat-labile enterotoxin produced were greater at initial pH 8.5 than at 7.0 of CYES-2 broth culture. However, the bacterial growth itself was more abundant at 7.0 than at 8.5. 2. Heat-labile enterotoxin per unit volume of culture supernatant was greater at shaking culture than at standing culture condition, but ratio of the enterotoxin produced over the unit mass of E. coli calculated was greater at standing culture than shaking culture condition, indicating that the greater yields of the toxin produced at shaking culture was due to increase in E. coli cell mass compared to the standing culture condition: 3. The enterotoxin produced in the lincomycin(128 microgram/ml) supplemented media was 5 or 11 times greater on the basis of enterotoxin per unit mass of E. coli, compared to the lincomycin-non-supplemented media, indicating that lincomycin itself increases the enterotoxin production. 4. Treatment of 18 hours culture of E. coli with polymyxin B(0.2 mg/ml) for 1 hour increased the yields of enterotoxin amounting to 2 or 5 times of the non-treated control cultures.

• Journal of Food Hygiene and Safety
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• v.6 no.3
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• pp.147-155
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• 1991

Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.1
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• pp.76-83
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• 1992

Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin (ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxigenic E. coli from non pathogenic E. coli. ST producing E. coli KM-7 strain was isolated from the swine and molecular cloning of ST gene of KM-7 strain. Transformant eKT-53 $(ST^+,\;LT^-)$ was selected by infant mouse assay (IMA). The culture supernatant of eKT-53 strain was performed purification by multipled steps. The culture supernatant (crude ST) was purified by sequentially applying batch adsorption chromatography on Amberlite XAD-2 resin, ion exchange chromatography on DEAE-Sephacel anion exchanger, gel filtration chromatography on Bio-Gel P-6 and preparative polyacrylamide slab gel electrophoresis. About 113-fold purification was achieved with a yield of about 11% of crude ST and the minimum effective dose(MED) of this purified ST was about 2.8ng in IMA. Homogeneity of purified ST was demonstrated by showing a single band in analytical SDS polyacrylamide disc gel electrophoresis.