• Parasites, Hosts and Diseases
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• v.44 no.1 s.137
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• pp.21-26
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• 2006

A shuttle vector for Escherichia coli and Giardia lamblia was modified to produce a reporter plasmid, which monitors the expression of prescribed gene in G. lamblia by measuring its luciferase activity. Promoter regions of the gap2 gene, one of the genes induced during encystation, were cloned into this plasmid, and the resultant constructs were then transfected into trophozoites of G. lamblia. Transgenic trophozoites containing one of the 3 gap2-luc reporters were induced to encystation, and characterized with respect to gap2 gene expression by measuring their luciferase activities. Giardia containing a gap2-luc fusion of 112-bp upstream region showed full induction of luciferase activity during encystation.

• 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.

• Parasites, Hosts and Diseases
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• v.54 no.1
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• pp.1-8
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• 2016

Laboratory workers, in resource-poor countries, still consider PCR detection of Giardia lamblia more costly and more time-consuming than the classical parasitological techniques. Based on 2 published primers, an in-house one-round touchdown PCR-RFLP assay was developed. The assay was validated with an internal amplification control included in reactions. Performance of the assay was assessed with DNA samples of various purities, 91 control fecal samples with various parasite load, and 472 samples of unknown results. Two cysts per reaction were enough for PCR detection by the assay with exhibited specificity (Sp) and sensitivity (Se) of 100% and 93%, respectively. Taking a published small subunit rRNA reference PCR test results (6%; 29/472) as a nominated gold standard, G. lamblia was identified in 5.9% (28/472), 5.2%, (25/472), and 3.6% (17/472) by PCR assay, $RIDA^{(R)}$ Quick Giardia antigen detection test (R-Biopharm, Darmstadt, Germany), and iodine-stained smear microscopy, respectively. The percent agreements (kappa values) of 99.7% (0.745), 98.9% (0.900), and 97.7% (0.981) were exhibited between the assay results and that of the reference PCR, immunoassay, and microscopy, respectively. Restriction digestion of the 28 Giardia-positive samples revealed genotype A pattern in 12 and genotype B profile in 16 samples. The PCR assay with the described format and exhibited performance has a great potential to be adopted in basic clinical laboratories as a detection tool for G. lamblia especially in asymptomatic infections. This potential is increased more in particular situations where identification of the parasite genotype represents a major requirement as in epidemiological studies and infection outbreaks.

• 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.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.429-433
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• 2000

Giardia lamblia, a human pathogen causing outbreaks of diarrhea, recently became a focus of great concerns in the fields of both medical and environmental microbioloty. To develop the experimental tools to study giardiasis, encystation, one of the major processes in its life cycle, was reconstituted by inducing an axenic culture of a flagellated form of G. lamblia into a cyst from under high concentration of bile and alkaline pH condition. The successful induction was confirmed by Northern analysis of resulting increased expression of the CWPl gene encoding the cyst wall protein 1. An examination of the encystation process with SEM (scanning electron microscopy) and TEM (transmission electron microscopy) revealed that the trophozoite, a flagellate with a bilateral symmetry, was transformed to a cyst form with an oval-shape and defined filamentous wall. The encystation was found to cause a disappearance of the flagella and an invagination of the adhesive disc. An extensive formation of rER (rough endoplasmic reticulum) was observed after 24h of induction, indication an active synthesis and export of proteins during this process. The vital staining of the invitro-induced systs showed that most cysts maintained their viability.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.2
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• pp.95-99
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• 2001

Giardia lamblia is a parasitic protozoa which is transmitted in the form of a cyst through untreated water and also treated drinking water. Since its presence in water has led to frequent outbreaks of giardiasis and death in many countries, the removal and disinfection of this protozoan cyst from the water supply are of great concern for public health. This study examined the disinfection characteristics of G. lamblia cysts isolated from a Korean patient with giardiasis. When using sodium hypochlorite including 5 or 10 ppm chlorine, the killing rate was initially rapid, however, the disinfection slowed down and a 3log reduction could not be achieved even after 2h. The disinfection effectiveness was also reduced at a lower temperature, thereby implying that the risk o a giardiasis outbreak will be higher in the winter season. A CT (concentration$.$time) curve was constructed based on the results with sodium hypochlorite for use in designing and predicting disinfection performance. The organic chlorination disinfectant SDIS (sodium dichloroisocyanurate) produced a lower pH and a much higher residual effect than sodium hypochlorite. The disinfection of cysts by SDIC continued steadily throughout 2h of contact, although the initial killing rate was lower than that with sodium hypochlorite.

• Parasites, Hosts and Diseases
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• v.60 no.5
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• pp.309-315
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• 2022

Inflammatory bowel disease (IBD) is a chronic and recurrent illness of the gastrointestinal tract. Treatment of IBD traditionally involves the use of aminosalicylic acid and steroids, while these drugs has been associated with untoward effects and refractoriness. The absence of effective treatment regimen against IBD has led to the exploration of new targets. Parasites are promising as an alternative therapy for IBD. Recent studies have highlighted the use of parasite-derived substances, such as excretory secretory products, extracellular vesicles (EVs), and exosomes, for the treatment of IBD. In this report, we examined whether EVs secreted by Giardia lamblia could prevent colitis in a mouse model. G. lamblia EVs (GlEVs) were prepared from in vitro cultures of Giardia trophozoites. Clinical signs, microscopic colon tissue inflammation, and cytokine expression levels were detected to assess the effect of GlEV treatment on dextran sulfate sodium (DSS)-induced experimental murine colitis. The administration of GlEVs prior to DSS challenge reduced the expression levels of pro-inflammatory cytokines, including tumor necrosis factor alpha, interleukin 1 beta, and interferon gamma. Our results indicate that GlEV can exert preventive effects and possess therapeutic properties against DSS-induced colitis.

• Parasites, Hosts and Diseases
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• v.54 no.3
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• pp.345-348
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• 2016

Although intestinal protozoans are common etiologies of diarrhea, few studies have been conducted in Myanmar. This study planned to investigate the prevalence of Giardia lamblia, Entamoeba coli, Entamoeba histolytica, and Endolimax nana among schoolchildren and their guardians in suburban areas near Yangon, Myanmar. We performed a cross-sectional survey among schoolchildren and their guardians from 7 primary schools in South Dagon and Hlaing Thar Yar districts, Yangon, Myanmar. Stool samples were observed with a microscope after concentration technique and iodine staining. Total 821 stool samples, including 556 from schoolchildren and 265 from guardians, were examined. The median age was 6 years old for schoolchildren and 36 years old for guardians. A 53.1% of the school children and 14.6 % of the guardians were males. The overall prevalence of each intestinal protozoan species was as follows: 3.4% (28/821) for G. lamblia; 3.5% (29/821) for E. coli; 1.2% (10/821) for E. histoytica, and 3.0% for E. nana. This study showed that intestinal protozoans are common in primary schoolchildren and their guardians in suburban areas near Yangon, Myanmar. Health interventions, such as hand washing education, improvement of sanitation, and establishment of water purification systems are urgently needed in this area.

• 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.23 no.2
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• pp.285-292
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• 1985

The present study has been designed as a basic study on laboratory diagnosis of giardiasis and to demonstrate a more effective method for the detection of Giardia lamblia cyst with the inherent advantages of minimizing both the number of stool examinations required and the interval of stool collections for estimating the real state of prevalence in the shortest time possible. There were 3 subject groups of 75 children each currently residing in an orphanage in Gunsan city, Jeonbuk province from which stool specimens were collected every day, every other day, and every 3 days. The procedure is as follows: 1) resuspend the InKed sample after fixation with SAF solution 2) centrifuge the sediment for 1 min. at 2, 000 rpm after straining through gauze into a tube 3) divide the sediment into 3 parts and use them for direct fecal smear, formalin-ether concentration (MGL) and zinc sulfate ($ZnSO_4$) floatation techniques. The results are summarized as follows: 1. Overall infection rate after 10 trials showed a 60% positive indication. The positive rate among children under 4 years old was significantly higher than the rate in children over 4 years old. No significant difference in rate by sex was observed. 2. The results of eBaminations by direct fecal smear and MGL techniques appeared more accurate than that obtained by $ZnSO_4$ ftoatation method as indicated by a higher positive rate. Of all three methods concerned, combinations of two demonstrated a higher positive rate than that shown by any one alone. 3. In three consecutive examinations under varying conditions such as different days, the cyst detection rate by MGL technique indicated 83%. In 5 examinations under the same varying conditions, the indicated rate was 94%. 4. The interval of stool collection proved to be insignificant for the cyst detection rate. In conclusion, both MGL method and modified fecal direct smear can provide a good cyst detection rate of G. lamblia provided that more than 3 consecutive examinations of stool under varying conditions are carried out.