• Parasites, Hosts and Diseases
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• v.56 no.1
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• pp.1-9
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• 2018

Giardia lamblia, an anaerobic, amitochondriate protozoan parasite causes parasitic infection giardiasis in children and young adults. It produces pyruvate, a major metabolic product for its fermentative metabolism. The current study was undertaken to explore the effects of pyruvate as a physiological antioxidant during oxidative stress in Giardia by cysteine-ascorbate deprivation and further investigation upon the hypothesis that oxidative stress due to metabolism was the reason behind the cytotoxicity. We have estimated intracellular reactive oxygen species generation due to cysteine-ascorbate deprivation in Giardia. In the present study, we have examined the effects of extracellular addition of pyruvate, during oxidative stress generated from cysteine-ascorbate deprivation in culture media on DNA damage in Giardia. The intracellular pyruvate concentrations at several time points were measured in the trophozoites during stress. Trophozoites viability under cysteine-ascorbate deprived (CAD) medium in presence and absence of extracellular pyruvate has also been measured. The exogenous addition of a physiologically relevant concentration of pyruvate to trophozoites suspension was shown to attenuate the rate of ROS generation. We have demonstrated that Giardia protects itself from destructive consequences of ROS by maintaining the intracellular pyruvate concentration. Pyruvate recovers Giardia trophozoites from oxidative stress by decreasing the number of DNA breaks that might favor DNA repair.

• 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.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.73-75
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• 2002

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.4
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• pp.368-371
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• 2006

The standard procedure outlined by the United States Environmental Protection Agency (US EPA) in Method 1623 for analyzing Giardia lamblia cysts and Cryptosporidium parvum oocysts in water samples consists of filtration, elution, centrifugal concentration, immunomagnetic separation (IMS), and immunofluorescence assay (IFA) followed by microscopic examination. In this study, the extent of (oo)cyst loss in each step of this procedure was evaluated by comparing recovery yields in segmented analyses: (i) IMS + IFA, (ii) concentration + IMS + IFA, and (iii) filtration/elution + concentration + IMS + IFA. The complete (oo)cyst recovery by the full procedure was $52{\sim}57%$. The (oo) cyst loss in the IMS step was only $0{\sim}6%$, implying that IMS is a fairly reliable method for (oo)cyst purification. Centrifugal concentration of the eluted sample and pellet collection before IMS resulted in a loss of $8{\sim}14%$ of the (oo)cysts. The largest (oo)cyst loss occurred in the elution step, with $68{\sim}71%$ of the total loss. The permeated loss of (oo)cysts was negligible during filtration of the water sample with a $1.0-{\mu}m$ pore polyethersulfone (PES) capsule. These results demonstrated that the largest fraction of (oo)cyst loss in this procedure occurred due to poor elution from the filter matrix. Improvements in the elution methodology are therefore required to enhance the overall recovery yield and the reliability of the detection of these parasitic protozoa.

• Korean Journal of Veterinary Research
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• v.50 no.4
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• pp.273-277
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• 2010

The objective of this study was to investigate the infection rate of gastro-intestinal tract parasites on acquired laboratory nonhuman primates, Vervet monkey, Cynomolgus monkey, and Rhesus monkey acquired from Japan and China. These monkeys have been acclimating at an individual housing condition after our legal quarantine period. We examined 133 fecal samples to investigate parasitic infection using direct smear and formalin-ether-sedimentation technique. As a result, total parasitic infection rate was 33.8% (n = 45/133) for all monkeys. Two species of macaques, cynomolgus and rhesus, were infected with Trichuris trichiura (4), Giardia lamblia (4) and Balantidium coli (41). Vervet monkeys, which had been controlled by individual housing system for a long time, were clear for parasitic infection. The protozoan, Balantidium coli was one of the most frequently detected in these monkey colonies. Double infection was noted in only 4 monkeys and involved with Trichuris trichiura and Balantidium coli. Serious clinical symptoms were not observed in the most of the infected monkeys, but the monkeys infected by Giardia lamblia showed intermittent or chronic watery diarrhea. Consequently, the prophylactic anthelmintic treatment and periodic monitoring are essential to preserve the SPF colonies in the laboratory facility.

• Journal of Microbiology and Biotechnology
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• v.14 no.6
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• pp.1105-1113
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• 2004

Disinfection of a waterborne pathogenic protozoa, Giardia lamblia, by the conventional chlorine method has been known to be difficult. An alternative disinfection has been carried out by using a UV -light illuminating optical­fiber photoreactor. Light intensity diffused from one piece of a clad-removed optical-fiber was $1- 1.5{\mu}Em^{-2}s^{-1}$. Disinfection capability in a UV -light irradiated optical-fiber reactor suspended with 0.01 g $TiO_{2}\;dm^{-3}$ was 1.4 times that in the same reactor without $TiO_{2}$ photocatalysts. To resolve the absorption and scattering of UV light by the particles themselves as well as the difficulty of recycling particles in the slurry­type reactor, $TiO_{2}$ which was obtained by a hydrothermal method, was immobilized on clad-removed optical fibers. Such pretreatment of fiber surface resulted in an excellent transparency, which enhanced the UV light to diffuse laterally from a fiber surface. Coating time of the prepared solution by the hydrothermal method was not effective after more than two times. Disinfection capability in the $TiO_{2}$-immobilized optical-fiber reactor was $83\%$ in 1 h at $40^{\circ}C$, which was slightly higher than $76\%$ at $22^{\circ}C$ and $68\%$ at $10^{\circ}C$. Disinfection capability at $22^{\circ}C$ increased from $74\%$ at an initial pH of 3.4, through $76\%$ at pH 6.5, to $87\%$ at an initial pH of 10. Oxygen supply with air-flow rate of 5 $cm^3\;min^{-1}$ did not seem to increase the disinfection capability with UV /immobilized $TiO_2$.

• Journal of Microbiology
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• v.41 no.2
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• pp.148-151
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• 2003

Waterborne parasite infections are considered a reemerging threat. Most studies on the epidemiology of human cryptosporidiosis, giardiasis, and amebiasis have been carried out in developed countries, and there is little data on the occurrence of these infections in other areas. The objective of this study was to investigate the presence of waterborne parasites such as Cryptosporidium parvum, Giardia lamblia and Entamoeba histolytica in various water samples in Ankara, turkey. A total of 85 samples were examined, 43 from the municipal water supply, 34 from wells, 6 from the Ankara River, and 2 from two untreated dams; by conventional microscopy, immunologically and by polymerase chain reaction (PCR). Oocysts of C. parvum and cysts of G. lamblia were detected by using an indirect fluorescence (antigen) assay, whereas an enzyme linked immunosorbent assay was used to detect the cysts of E. histolytica and E. dispar. In addition, PCR was used for E. histolytica, E. dispar, C. parvum and G. lamblia detection. G. lamblia was found in 2 of the 34 well water samples, and parasites were found in 3 of the 6 Ankara River samples. The 1$\^$st/ contained E. histolytica cysts and Strongyloides stercoralis larvae. the 2$\^$nd/ E. histolytica cysts, and Trichuris trichiura eggs, and the 3$\^$rd/ C. parvum oocysts only. No parasite was observed in the municipal water samples and untreated dam water samples. These results extend our knowledge on waterborne parasites, such occurrence information on waterborne pathogens assists the management and treatment of municipal water.

• Proceedings of the Microbiological Society of Korea Conference
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• 2003.05a
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• pp.105-107
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• 2003

• Parasites, Hosts and Diseases
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• v.47 no.3
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• pp.303-305
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• 2009

The incidence and etiology of parasite-associated gastroenteritis during 2004-2006 in Gyeonggi-do (province), South Korea was determined by means of antigen detection ELISA on 6,071 stool specimens collected from 6 general hospitals. At least 1 parasitic agent was detected in 3.4% (208/6,071) of the stool samples. Among these, Giardia lamblia was the most numerous (152 cases; 2.5%), followed by Entamoeba histolytica (25 cases; 0.4%), Cryptosporidium parvum (23 cases; 0.4%), and mixed infections (8 cases; 0.1%). Patients aged 1-5years had the largest proportion (69.2%; 144/208) of parasite-positive stool specimens. Parasite-mediated gastroenteritis was most common from June to September. The detection rate gradually increased from 2004 to 2006. This study shows that parasite-mediated gastroenteritis may be significant among children in Korea and that parasite infection surveillance should be constantly performed.

• Parasites, Hosts and Diseases
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• v.51 no.5
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• pp.519-524
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• 2013

This study investigated the effect of breast-feeding in protection against protozoan infection in infants with persistent diarrhea. Infants were classified into 2 groups; 161 breast-fed infants and the same number of non-breast-fed infants. Microscopic examinations of stool were done for detection of parasites and measuring the intensity of infection. Moreover, serum levels of IgE and TNF-${\alpha}$ were measured by ELISA. Cryptosporidium spp., Entamoeba histolytica/Entamoeba dispar, Giardia lamblia, and Blastocystis sp. were demonstrated in infants with persistent diarrhea. The percentage of protozoan infections was significantly lower in breast-fed infants than that in the non-breast-fed infants. The levels of IgE and TNF-${\alpha}$ were significantly lower in the breast-fed group than in the non-breast-fed group. There were significant positive associations between the serum levels of IgE and TNF-${\alpha}$ and the intensity of parasite infection in the breast-fed group. It is suggested that breast-feeding has an attenuating effect on the rate and intensity of parasite infection.