• 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

• Proceedings of the Korean Society of Veterinary Service Conference
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• 1997.05a
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• pp.244.2-245
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• 1997

• Journal of Korean Society of Water and Wastewater
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• v.20 no.1
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• pp.71-78
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• 2006

The removal of protozoa in the coagulation process was evaluated under the different pH and turbidity using the jar test after the addition of polyaluminium chloride (PAC) as a coagulant. Two well-known protozoa of Cryptosporidium parvum and Giardia lamblia were tested at the same time with turbidity, the critical water quality parameter of the water treatment process. Both protozoa were removed about 1log (and up to 2log) at the optimum injection of PAC. The source water turbidity and pH affected the removal of protozoa and turbidity. At neutral and alkaline pH, 1.3-1.7log removal of protozoa for low turbid water with 5NTU, and 1.6-2.3log removal for high turbid water with 30NTU were achieved. However, at acidic pH, maximum 0.8-1.0log and 1.1-1.2log were removed for low and high turbid water, respectively, at the optimum PAC injection of 15mg/L. The relation of protozoa and turbidity removals were expressed as the 1st order equation (significantly positive relation) in the most of the tested conditions. In addition, the relation of protozoan removals with residual turbidity were also expressed the 1st order equation (significantly negative relation), although the significance of the equations were reduced at acidic pH. Therefore, residual turbidity could be a good index of efficient protozoan removal in the coagulation process, probably except at the low pH condition.

• 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 $TiO_2$.

• Parasites, Hosts and Diseases
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• v.51 no.2
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• pp.237-241
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• 2013

Giardia lamblia is recognized as one of the most prevalent parasites in dogs. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of G. lamblia from dogs. The fecal samples were collected and prepared for microscopic analysis, and then the genomic DNA was extracted directly from purified cysts. The concentration of DNA samples of G. lamblia were diluted by 10-fold serially ranging from $10^{-1}$ to $10^{-5}ng/{\mu}l$ for LAMP and PCR assays. The LAMP assay allows the amplification to be finished within 60 min under isothermal conditions of $63^{\circ}C$ by employing 6 oligonucleotide primers designed based on G. lamblia elongation factor 1 alpha ($EF1{\alpha}$) gene sequence. Our tests showed that the specific amplification products were obtained only with G. lamblia, while no amplification products were detected with DNA of other related protozoans. Sensitivity evaluation indicated that the LAMP assay was sensitive 10 times more than PCR. It is concluded that LAMP is a rapid, highly sensitive and specific DNA amplification technique for detection of G. lamblia, which has implications for effective control and prevention of giardiasis.

• Parasites, Hosts and Diseases
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• v.53 no.1
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• pp.119-124
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• 2015

The objective of this study was to genetically characterize isolates of Giardia duodenalis and to determine if zoonotic potential of G. duodenalis could be found in stray cats from urban and suburban environments in Guangzhou, China. Among 102 fresh fecal samples of stray cats, 30 samples were collected in Baiyun district (urban) and 72 in Conghua district (suburban). G. duodenalis specimens were examined using light microscopy, then the positive specimens were subjected to PCR amplification and subsequent sequencing at 4 loci such as glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi), ${\beta}$-giardin (bg), and small subunit ribosomal RNA (18S rRNA) genes. The phylogenetic trees were constructed using obtained sequences by MEGA5.2 software. Results show that 9.8% (10/102) feline fecal samples were found to be positive by microscopy, 10% (3/30) in Baiyun district and 9.7% (7/72) in Conghua district. Among the 10 positive samples, 9 were single infection (8 isolates, assemblage A; 1 isolate, assemblage F) and 1 sample was mixed infection with assemblages A and C. Based on tpi, gdh, and bg genes, all sequences of assemblage A showed complete homology with AI except for 1 isolate (CHC83). These findings not only confirmed the occurrence of G. duodenalis in stray cats, but also showed that zoonotic assemblage A was found for the first time in stray cats living in urban and suburban environments in China.

• Parasites, Hosts and Diseases
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• v.53 no.4
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• pp.477-481
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• 2015

Giardia is a major public health concern and considered as reemerging in industrialized countries. The present study investigated the prevalence of giardiosis in 202 sheltered dogs using PCR. The infection rate was 33.2% (67/202); Gyeongsangbuk-do and Daejeon showed 25.7% (39/152, P<0.0001) and 56% (28/50), respectively. The prevalence of infected female dogs (46.7%, P<0.001) was higher than in male dogs (21.8%). A higher prevalence (43.5%, P<0.0001) was observed in mixed breed dogs than purebred (14.1%). Although most of the fecal samples collected were from dogs of ${\geq}1$ year of age which showed only 27.4% positive rate, 61.8% (P<0.001) of the total samples collected from young animals (<1 year of age) were positive for G. intestinalis. A significantly higher prevalence in symptomatic dogs (60.8%, P<0.0001) was observed than in asymptomatic dogs (23.8%). Furthermore, the analysis of nucleotide sequences of the samples revealed that G. intestinalis Assemblages A and C were found in the feces of dogs from Gyeongsangbuk-do and Daejeon. Since G. intestinalis Assemblage A has been known to infect humans, our results suggest that dogs can act as an important reservoir of giardiosis in Korea. Hence, hygienic management should be given to prevent possible transmission to humans.

• Parasites, Hosts and Diseases
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• v.52 no.3
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• pp.263-271
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• 2014

PCR detection of intestinal protozoa is often restrained by a poor DNA recovery or by inhibitors present in feces. The need for an extraction protocol that can overcome these obstacles is therefore clear. $QIAamp^{(R)}$ DNA Stool Mini Kit (Qiagen) was evaluated for its ability to recover DNA from oocysts/cysts directly from feces. Twenty-five Giardia-positive, 15 Cryptosporidium-positive, 15 Entamoeba histolytica-positive, and 45 protozoa-free samples were processed as control by microscopy and immunoassay tests. DNA extracts were amplified using 3 sets of published primers. Following the manufacturer's protocol, the kit showed sensitivity and specificity of 100% towards Giardia and Entamoeba. However, for Cryptosporidium, the sensitivity and specificity were 60% (9/15) and 100%, respectively. A series of optimization experiments involving various steps of the kit's protocol were conducted using Cryptosporidium-positive samples. The best DNA recoveries were gained by raising the lysis temperature to the boiling point for 10 min and the incubation time of the InhibitEX tablet to 5 min. Also, using a pre-cooled ethanol for nucleic acid precipitation and small elution volume ($50-100{\mu}l$) were valuable. The sensitivity of the amended protocol to Cryptosporidium was raised to 100%. Cryptosporidium DNA was successfully amplified by either the first or the second primer set. When applied on parasite-free feces spiked with variable oocysts/cysts counts, ${\approx}2$ oocysts/cysts were theoretically enough for detection by PCR. To conclude, the Qiagen kit with the amended protocol was proved to be suitable for protozoan DNA extraction directly from feces and support PCR diagnosis.