• Microbiology and Biotechnology Letters
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• v.33 no.2
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• pp.148-153
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• 2005

Malaria is a re-emerging infectious disease that is spreading to areas where it had been eradicated, such as Eastern Europe and Central Asia. To avoid the mortality from malaria, early detection of the parasite is a very important issue. The peripheral blood smear has been the gold standard method for the diagnosis of malaria infection. Recently, several other methods have been introduced for quantitative detection of malaria parasites. Real time PCR that employs fluorescent labels to enable the continuous monitoring of PCR product formation throughout the reaction has recently been used to detect several human malaria parasites. 18S rRNA sequences from malaria parasites have been amplified using Taqman real time PCR assay. Here, a SYBR Green-based real time quantitative PCR assay for the detection of malaria parasite-especially, Plasmodium vivax - was applied for the evaluation of 26 blood samples from Korean malaria patients. Even though SYBR Green-based real time PCR is easier and cheaper than Taqman-based assay, SYBR Green-based assay cannot be used because 18S rRNA cannot be specifically amplified using 1 primer set. Therefore, we used DBP gene sequences from Plasmodium vivax, which is specific for the SYBR Green based assays. We amplified the DBP gene from the 26 blood samples of malaria patients using SYBR Green based assay and obtained the copy numbers of DBP genes for each sample. Also, we selected optimal reference gene between ACTB and B2M using real time assay to get the stable genes regardless of Malaria titer. Using selected ACTB reference genes, we successfully converted the copy numbers from samples into titer, ${\sharp}$ of parasites per microliter. Using the resultant titer from DBP based SYBER Green assay with ACTB reference gene, we compared the results from our study with the titer from Taqman-based assay. We found that our results showed identical tendency with the results of 18S rRNA Taqman assay, especially in lower titer range. Thus, our DBP gene-utilized real time assay can detect Plasmodium vivax in Korean patient group semi-quantitatively and easily.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.357-364
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• 1995

The protozoan parasite, Entcmoeba histoIWticc, is one of major causative agents of intestinal disease all over the world. In acute experimental infection, the early host response to 5. histoIHtica is characterized by an infiltration of neutrophils. However, the chemotactic signal for this response is not well known. Based on the (jading that human epithelial cells produce the potent neutrophil chemoattractant and activator, interleukin-8 (IL-8), IL-8 gene expression was examined thoroughly in human colon epithelial cells exposed to 5. histolvtica trophozoites. Cellular RNAs were extracted from HT-29 or Caco-2 human colon epithelial cells exposed to 5. histoLvtica trophozoites for 30 minutes, 1 and 3 hours. IL-8 mRNA transcripts were measured by reverse transcriptional polprnerase chain reaction (RT-PCR) using synthetic standard RNA. The number of IL-8 mRNA molecules increased from 30 minutes to 3 hours of exposure period, reaching 3.1 H 107 molecules/ug of total RNA. Expression pattern of IL-8 mRNA transcripts was parallel to the amounts of IL-8 protein measured by enzyme-linked immunosorbent assay (ELISA) . Lysates of 5. histoIVtica also induced expression of mRNA for IL-8 in colon epithelial cells. These results sugf:esc that acute inflammatory reaction by 5. histoIVticc may be initially triggered by proinflammatory cytokines such as IL-8 secreted from epithelial cells of the colon.

• Journal of fish pathology
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• v.26 no.2
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• pp.65-75
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• 2013

Population growth equation of scuticociliate Miamiensis avidus was obtained from the experimental results of in vitro culture condition to estimate the growth rate and carrying capacity from the growth equation. In addition, intraperitoneal infections into olive flounder Paralichthys olivaceus were carried out into 2 different conditions: different concentrations of M. avidus in same water temperature and same concentration of M. avidus in different water temperatures. Olive flounder mortality was threshold dependent with both the temperature and M. avidus density parameters. In this paper, we propose a mathematical model to study M. avidus growth in olive flounder based upon the interactions between parasite and host. The mathematical model was logistic growth differential equation (1.2). The parameters were found with Matlab program through the Levenberge-Marquardt method. In theorem, equilibrium values between the infected fish population and dead population could found. Our equilibrium points were a stable equilibrium and an unstable equilibrium. From the equation (1.6), it was possible to predict the amount of cumulative mortality of olive flounder along with the time after M. avidus infection.

• Korean Journal of Veterinary Service
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• v.35 no.4
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• pp.339-342
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• 2012

The objective of this study was to determine the infection patterns of etiological agents causing calf diarrhea in the Gyeongnam province, Korea. In this study, from January 2011 to December 2011, feces and necropsy specimens from 249 calves diagnosed with diarrhea (<7 months old) were examined by reverse transcriptase-polymerase chain reaction assay and bacteria & coccidium isolation for detection pathogenic organism. The results of this study showed that 78 cases (31.3%) in spring, 71 cases (28.5) in summer, 62 cases (24.9%) in fall and 38 cases (15.3%) in winter were diagnosed with calf diarrhea, respectively. Calf diarrhea-causing pathogens were diagnosed as bacteria 113 (45.4%), viruses 97 (39.0%), coccidium 1 (0.4%), unknown cases 13 (5.2%), and mixed infections 25 (10.0%). We isolated three virus types from fecal samples (97), which were classified as BVD 64 (66.0%), BRV 21 (21.6%), and BCV 12 (12.4%). Moreover, co-infected pathogens were 25 cases, consisting with BVD & BRV 11 (44%), BVD & BCV& BRV 7 (28.0%), E. coli & BCV 3 (12%), and BVD & IBR 1 (4.0%). In summary, we demonstrated that the enteropathogens of bacteria, viruses, and parasite were detected in samples from cattle with diarrhea, principally in young calves less than 7 months of age. Future studies of infectious diarrhea in cattle should include assays for this etiologic agent.

• The Journal of the Korean Society for Microbiology
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• v.34 no.6
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• pp.519-532
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• 1999

Helicobacter pylori is a causative agent of type B gastritis and plays a central role in the pathogenesis of gastroduodenal ulcer and gastric cancer. To elucidate the host-parasite relationship of the H. pylori infection on the basis of molecular biology, we tried to evaluate the genomic diversity of H. pylori. An ordered overlapping bacterial artificial chromosome (BAC) library of a Korean isolate, H. pylori 51 was constructed to set up a genomic map. A circular physical map was constructed by aligning ApaI, NotI and SfiI-digested chromosomal DNA. When the physical map of H. pylori 51 was compared to that of unrelated strain, H. pylori 26695, completely different restriction patterns were shown. Fifteen known genes were mapped on the chromosome of H. pylori 51 and the genetic map was compared with those of strain 26695 and J99, of which the entire genomic sequences were reported. There were some variability in the gene location as well as gene order among three strains. For further analysis on the genomic diversity of H. pylori, when comparing the genomic structure of 150 H. pylori Korean isolates with one another, genomic macrodiversity of H. pylori was characterized by several features: whether or not susceptible to restriction digestion of the chromsome, variation in chromosomal restriction fingerprint and/or high frequency of gene rearrangement. We also examined the extent of allelic variation in nucleotide or deduced amino acid sequences at the individual gene level. fucT, cagA and vacA were confirmed to carry regions of high variation in nucleotide sequence among strains. The plasticity zone and strain-specific genes of H. pylori 51 were analyzed and compared with the former two genomic sequences. It should be noted that the H. pylori 51-specific sequences were dispersed on the chromosome, not congregated in the plasticity zone unlike J99- or 26695-specific genes, suggesting the high frequency of gene rearrangement in H. pylori genome. The genome of H. pylori 51 shows differences in the overall genomic organization, gene order, and even in the nucleotide sequences among the H. pylori strains, which are far greater than the differences reported on the genomic diversity of H. pylori.

• Journal of fish pathology
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• v.10 no.1
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• pp.21-29
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• 1997

We investigated on the prevalence and extermination of scuticociliatids parasitic on cultured japanese flounder, Paralichthys olivaceus in land-marine tank system of southern Korea from January to February in 1997. The gills and the skin showed the highest infection rate(60%), and the brain showed the lowest(22%). Also, fish secreted large quantity of mucus with a bleeding and ulcerated lesions on the infected sites. The number of the parasites in inflowing sea water, surface water and bottom water of farming tank ranged 0~1 individuals/$100m\ell$, 0~413 individuals/$100m\ell$ and $7\sim7.3{\times}10^4$ individuals/$100m\ell$, respectively. This parasite was died within 2 hours in 50~500 ppm, 48 hours of 10 ppm formalin or hydrogen peroxide, 1 hour in 50~500 ppm, 80 minutes of 10 ppm oligo chitosan and 10 minute in 100% but did not died until 48 hours in 10~70% fresh water.

• Parasites, Hosts and Diseases
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• v.27 no.4
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• pp.253-260
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• 1989

Metacercariae of the genus Stictodora encysted in the head tissue of Acanthogobius navimanus (the gobies) caught at Sachun-gun, Kyongnam Province, were identified to be Stictodora Zari Yamaguti, 1939 (Trematoda: Heterophyidae), a new parasite fauna in Korea. The metacercariae were 0.39∼,0.43 mm by 0.32∼0.35 mm in size, long elliptical, and with a thin and transparent cyst wall. Total 200 metacercariae were collected from 50 gobies. In order to obtain adult worms two kittens and a Puppy were infected each with 34∼100 metacercariae, and total 33 adults were recovered between the day 4 and day 8 post-infection. The S. sari adults measured 0.95∼1.18 mm long and 0.26∼0.32 mm wide and the eggs in uteri 0.028∼0.033 mm by 0.017∼0.020 mm. The most characteristic morphological feature of these flukes was the presence of a gonotyl and gonotyl spines arranged in two groups; densely crowded group of 30~40 spines and linearly-arranged one of 30∼40 spines, together of which made a comma(or reversed comma) shape along the lateral margin of the gonotyl. It has been proved by this study that 5. sari is distributed in southern coasts of Korea.

• Parasites, Hosts and Diseases
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• v.54 no.3
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• pp.253-259
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• 2016

In the era of (pre) elimination setting, the prevalence of malaria has been decreasing in most of the previously endemic areas. Therefore, effective cost- and time-saving validated pooling strategy is needed for detection of malaria in low transmission settings. In this study, optimal pooling numbers and lowest detection limit were assessed using known density samples prepared systematically, followed by genomic DNA extraction and nested PCR. Pooling strategy that composed of 10 samples in 1 pool, $20{\mu}l$ in 1 sample, was optimal, and the parasite density as low as $2p/{\mu}l$ for both falciparum and vivax infection was enough for detection of malaria. This pooling method showed effectiveness for handling of a huge number of samples in low transmission settings (<9% positive rate). The results indicated that pooling of the blood samples before DNA extraction followed by usual nested PCR is useful and effective for detection of malaria in screening of hidden cases in low-transmission settings.

• 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.28 no.4
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• pp.213-220
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• 1990

The employees at the Pohang industrial area, where Clonarchis sinensis has been known to be endemic along the Hyungsan River, were examined parasitologically for clonorchiasis and a part of the infected cases were surveyed with a questionaire to outline the recent infection status of C. sinensis and epidemiological parameters in the area. Total of 3, 180 cases were tested by intra- dermal inoculation of C. sinensis antigen (Green Cross Co., Korea), and 834(26.2%) were found positive. Out of the positive cases, 598 were subjected to (ecal examination for helminth ova. The examination revealed 129(21.6%) ova Positive cases of C. sinensis, and Trichuris trichiura 1.7%, Ascaris lumbricoides 0.3%, and Metagonimus yokogawai 0.2%. The questionaire analysis showed some significant difFerences between the infected and non-infected(control) groups. The infected cases were less educated than the control, and they lived at the closer area to the river, and most of them lived there over 20 years. Also they preferred eating raw fresh water fish. Most of the detected cases were treated with prasiquantel and found negative for the eggs in 85.3% of them 1 year after the treatment. The present data reveal markedly decreased endemicity of clonorchiasis compared with previous prevalence rates but still clonorchiasis is endemic in the Hyungsan river basin. A comprehensive measure including case detection, treatment and education for parasite control should be applied to control clonorchiasis in such endemic areas.