Lee, Hye Jin;Kim, Ha Nui;Yoo, Byong Joon;Kim, Jang Su;Kim, Myong Han;Lim, Chae Seung;Lee, Kap No
Laboratory Medicine Online
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v.1
no.2
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pp.100-104
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2011
Background: Malaria is a problematic disease in Korea, and microscopic examination of Giemsa-stained blood smear has been used as the gold standard for its diagnosis. However, this technique is time-consuming and has low sensitivity in samples with low numbers of malarial parasites (<20 parasites/μL). Here, we evaluated the performance characteristics of the LG AdvansureTM Malaria P.f./P.v. real-time QPCR (LG life sciences, Korea). Methods: Blood samples from 173 persons who visited Korea University Ansan Hospital were evaluated. QPCR was performed in 73 malaria patients and 100 healthy subjects by using the LG Advansure Malaria P.f./P.v. real-time QPCRR kit, and the results were compared with those of microscopy. The detection limit of this kit was determined by serial dilution of Plasmodium-infected blood with normal blood (blood not infected with Plasmodium). Results: Among the 73 patients that were microscopically confirmed to have malaria (Plasmodium vivax infection, N=70, P. falciparum infection, N=3), 69 patients were diagnosed with P. vivax infection and 3 were diagnosed with P. falciparum infection by LG AdvansureTM Malaria P.f./P.v. realtime QPCR. Both the tests indicated absence of infection in the 100 healthy subjects. The detection limit of LG AdvansureTM Malaria P.f./P.v. real-time QPCR was 0.1 parasite/μL. Conclusions: LG AdvansureTM Malaria P.f./P.v. real-time QPCRis a very sensitive and specific technique and can be used as a confirmatory test for malaria.
Kim, Jin Hui;Song, Jun Young;Lee, Jung-Ho;Hur, Jun Wook;Kwon, Se Ryun;Kwon, Joon Yeong
Korean Journal of Ecology and Environment
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v.52
no.4
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pp.306-315
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2019
The estuary of the Han River constantly suffers from pollutants and pathogenic microorganisms which could cause serious damage to aquatic organisms living there. Despite of this potential risk, it is hard to find any reliable scientific reports on the status of reportable disease infection to the organisms living in this area. In this study, cyprinid fish and crustaceans in Jeonryu-ri, a region of the Han River estuary, were investigated for the infection by representative reportable communicable diseases(SVC, spring viraemia of carp; KHVD, koi herpesvirus disease; EUS, epizootic ulcerative syndrome; WSD, white spot disease) and parasites. Peripheral fish and primary freshwater fish were observed in Jeonryu-ri with cyprinid caught most frequently. Crustaceans were mostly marine species. No positive bands to any of the reportable diseases were produced in any of the fish and crustacean examined in this study by PCR. No trace of Clonorchis sinensis, a liver fluke potential threat to human health, was detected in any of fish samples. However, many fish were infected by metacecaria of other flukes, and other various parasites such as nematode, cestode, copepod, monosite and acanthocephalan. These results suggest that important aquatic organisms in the Han River estuary is not seriously polluted yet. However, it is important to keep monitoring the diseases since the water quality in this region is constantly changing, and devastating influence of infectious diseases is unpredictable. Further, it is required to expand monitoring area toward upstream and increase the number of fish for examination.
Infections of Toxoplasma gondii and Babesia microti are reported in many wild animals worldwide, but information on their incidence and molecular detection in Korean wild fields is limited. In this study, the prevalence of T. gondii and B. microti infection in blood samples of 5 animal species (37 Chinese water deer, 23 raccoon dogs, 6 roe deer, 1 wild boar, and 3 Eurasian badgers) was examined during 2008-2009 in Gangwon-do (Province), the Republic of Korea (=Korea) by using serological and molecular tests. The overall seropositivity of T. gondii was 8.6% (6/70); 10.8% in Chinese water deer, 4.3% in raccoon dogs, and 16.7% in roe deer. PCR revealed only 1 case of T. gondii infection in Chinese water deer, and phylogenic analysis showed that the positive isolate was practically identical to the highly pathogenetic strain type I. In B. microti PCR, the positive rate was 5.7% (4/70), including 2 Chinese water deer and 2 Eurasian badgers. Phylogenetic analysis results of 18S rRNA and the ${\beta}$-tubulin gene showed that all positive isolates were US-type B. microti. To our knowledge, this is the first report of B. microti detected in Chinese water deer and Eurasian badger from Korea. These results indicate a potentially high prevalence of T. gondii and B. microti in wild animals of Gangwon-do, Korea. Furthermore, Chinese water deer might act as a reservoir for parasite infections of domestic animals.
Nosema spp. (NS) causes severe problems to honey bee colonies including the death of the highly infected ones. Searching for effective materials to control this parasite is very important. The objectives of this study were to identify the calendar for the prevalence of NS and other bee diseases, and to test the efficacy of three materials: diluted honey mixed with lemon juice (M1), chamomile extract mixed with sugar syrup (M2) and sutrivet mixed with sugar syrup (M3) against Nosema. To realize these objectives, diseases of brood and adult honey bees were surveyed over one year. Also, the efficacy of M1, M2 and M3 against Nosema was evaluated under field and laboratory conditions. The results showed that few diseases for immature and mature stages of honey bees were recorded. NS was detected during winter and spring in link with low temperature and high relative humidity. Under field conditions, M2 reduced the infection by 36.66% while M3 by 23.33% and finally M1 by 13.33%. In the laboratory, the highest efficacy was to M2 followed by M1 and finally M3. The three materials impacted the percentage of survived bees significantly higher than infected bees without any treatments over the experimental period. The study suggests the potential role of chamomile as a natural material to control NS.
Pneumonia is an infection of the lungs and respiratory system and can be classified by a variety of factors such as infectious agents, etiology, infection area, and other criteria. From a 46-year-old male, who was suspected of being infected with atypical pathogen pneumonia and underwent such tests as serological testing, examination of sputum, urine examination, parasite examination, bronchoscopy, needle biopsy and so on, no significant abnormality was found. This patient also showed no specific symptoms like auscultatory abnormalities, high fever, nonproductive cough, muscle stiffness, sputum production, dyspnea. Prescription of broad-spectrum oral antibiotics and ant-parasitic didn't seem to be effective against bacterial and atypical pathogen. The patient's condition alternately repeated between natural cure and recurrence. The average healing process during which scarring, nodule recurrence and disappearance on the lungs happened was about 20 days. Chest radiography and chest high resolution computerized tomographic scans(HRCT scan) was performed to depict parenchymal aberrations and demarcate the extent and distribution of atypical pathogen pneumonia. As a result, chest radiography did not show the specific symptoms, whereas areas of opacity (seen as white) which represent consolidation were revealed in chest HRCT scan. This indicates that only chest radiography is not that useful for early diagnosis of atypical pathogen pneumonia in patients, since it can't show exactly what the symptom is because of the barriers such as diaphragm, liver, and spine. Therefore, it is desirable that chest HRCT should be used in the diagnosis to compare with the results of chest radiography. Here, report with literature investigations the case of recurrent atypical pathogen pneumonia.
Kim, Jung-Mi;Song, Ha-Yeon;Yun, Suk-Hyun;Lee, Hyun-Suk;Ko, Han-Kyu;Kim, Dae-Hyuk
한국균학회소식:학술대회논문집
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2015.11a
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pp.37-37
/
2015
dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.
Fibroblasts are the most common cells in connective tissue and are responsible for the synthesis of extracellular matrix components. The fibrosis associated with chronic inflammation and injury may contribute to cholangiocarcinoma pathogenesis, particularly through an increase in extracellular matrix components, which participate in the regulation of bile duct differentiation during development. Mitochondria produce ATP through oxidative metabolism to provide energy to the cell under physiological conditions. Also, mitochondrial dysfunction and oxidative stress have been implicated in cellular senescence and aging. Alternations in mitochondrial structure and function are early events of programmed cell death or apoptosis and mitochondria appear to be a central regulator of apoptosis in most somatic cell. Clonorchis sinensis, one of the most important parasite of the human bile duct in East Asia, arouses epithelial hyperplasia and ductal fibrosis. Isolated fibroblast from the bile ducts of rats infected by C. sinensis showed increase of cytoplasmic process. In addition, decrease of cellular proliferation was observed in fibroblasts which was isolated from normal rat bile duct and then cultured in media containing C. sinensis excretory-secretory product. However, the effects of C. sinensis infection on the mitochondrial enzyme distribution is not clearly reported yet. Therefore, we investigated the structural change of C. sinensis infected bile duct and mitochondrial enzyme distribution of the cultured fibroblast isolated from the C. sinensis infected rat bile duct. As a result, C. sinensis infected SD rat bile ducts showed the features of chronic clonorchiasis, such as ductal connective and epithelial tissue dilatation, or ductal fibrosis. In addition, fibroblast in ductal connective tissue was damaged by physical effect of fibrotic tissue and chemical stimulation. Immunohistochemically detected mitochondrial electron transferase (ATPase, COXII, Porin) was decreased in C. sinensis infected rat bile duct and cultured fibroblast from infected rat bile duct. It can be hypothesized that the reason why number of electron transferase decrease in fibroblast isolated from the rat bile duct infected with C. sinensis is because dysfunction of electron transport system is occurred mitochondrial dysfunction, increase of ROS (reactive oxygen species) and apoptosis after chemical damage on the cell caused by C. sinensis infection. Overall, C. sinensis infection induces fibrotic change of ductal connective tissue, mutation of cellular metabolism in fibroblast and mitochondrial dysfunction. Consequently, ductal fibrosis inhibits fibroblast proliferation and decreases mitochondrial electron transferase on fibroblast cytoplasm. It was assumed that the structure of bile duct could not normalized and ductal fibrosis was maintained for a long period of time according to fibroblast metamorphosis and death induced by mitochondrial dysfunction.
The present status of control measures for public health important helminthic infections in Korea was surveyed in 1969 and the following results were obtained. The activities of parasitic examination and Ascaris treatment for the positives which were done during 1966 to 1969 were brought in poor result and could not decrease the infection rate. It is needed to improve or strengthen the activities. The mass treatment activities for paragonimiasis and clonorchiasis in the areas which were designated by the Ministry of Health were carried out during 1965 to 1968 with no good results in decrease of estimated number of the patients. There were too many pharmaceutical companies where many kinds of anthelmintics were produced. It may be better to reduce the number of anthelmintics produced and control the quality. The human feces, the most important source of helminthic infections, was generally not treated in sanitary ways because of the poor sewerage system and no sewage treatment plant in urban areas and insanitary latrines in rural areas. The field soils of 170 specimens were collected from 34 areas out of 55 urban and tourist areas where night soil has been prohibited by a regulation to be used as a fertilizer, and examined for parasites contamination with the result of Ascaris egg detection in 44%. Some kinds of vegetables of 64 specimens each from the supply agents of parasite free vegetables and general markets were collected and examined for parasites contamination with the results of Ascaris egg detection in 25% and 36% respectively. The parasite control activities and the ability of parasitological examination techniques in the health centers of the country were not satisfactory. The budget of the Ministry of Health for the parasite control was very poor. The actual expenditure needed for cellophane thick smear technique was 8 Won per a specimen. As a principle the control of helminthic infections might be led toward breaking the chain of events in the life cycle of the prasites and eliminating environmental and host factors concerned with the infections, and the following methods nay be pointed out. 1) Mass treatment might be done to eliminate human reservoirs of an infection. 2) Animal reservoirs which are related with human infections night be eliminated. 3) The excretes of reservoirs, particularly human feces, should be treated in sanitary ways by the means of sanitary sewerage system and sewage treatment plant in urban areas and sanitary latrines such as waterborne latrine, aqua privy and pit latrine in rural areas. The increase of national economical development and prohibition of the habit of using night soils as a fertilizer might be very important factors to achieve the purpose. 4) The control of vehicles and intermediate hosts might be done by the means of prohibition of soil contamination with parasites, food sanitation, insect control and snail control. 5) The improvement of insanitary attitudes and bad habits which are related with parasitic infections night be done by the means of prohibition of habit of using night soils as a fertilizer, and improving eating habits and personal hygiene. 6) Chemoprophylactic measure and vaccination may be effective to prevent the infections or the development of a parasite to adult in the bodies when the bodies were invaded by parasites. Further studies and development of this kind of measures are needed.
As the consumption of environmentally friendly agricultural products increases, food safety is at the forefront of public health concerns. We analyzed the biological hazards of 26 species of leaf vegetables and 4 species of fruit according to types of cultivation (conventional or organic filming) and distribution system (giant retailers or organic food stores) using various culture media, automatic bacterial identification systems, and microscopy, Total bacterial count of unwashed agricultural product ranged from $5.2{\times}10^{3}\;to\;1.5{\times}10^{5}\;CFU/mL$ (from 0.1 g of agricultural products), and the average count dropped 25-fold (range, 8-60-fold) after water washing. Microbial levels of washed organic agricultural products were $6.0{\times}10^{2}-1.9{\times}10^{4}\;CFU/mL$, and were not significantly different f개m the microbial loads on conventionally farmed produce. There was no significant difference in bacterial count from agricultural produce purchased from giant retailers or organic food stores. Total microbial count of Chinese cabbage, welsh onion, red chicory and kale were comparatively high, and Enterobacter cloacae was isolated most frequently. Parasites were detected in agricultural products purchased from conventional farm products in the stores of giant retailers, and in organic food stores, and parasite prevalence was especially high in Chinese cabbages and welsh onion. The study indicated that cultivation methods and distribution systems did not cause significant differences in biological contamination levels of agricultural produce. Some vegetables and fruits were highly contaminated effective sanitizing methods to reduce these biological hazards are needed.
A pathogenic free-living amoeba, Naegleria fowleri, is a causative protozoan parasite of primary amebic meningoencephalitis in human and experimental animals. It is known that humoral and cellular immunity contribute as the defence mechanism of host against this organism. Recently splenectomy has been argued on its effect on host defence mechanisms. The present study was aimed to observe the enact of immunization in splenectomized mice. For immunization, $5~10{\times}10^5$ trophozoites of Naegleria fewleri o 359 were intraperitoneally inoculated once a week for two weeks to BALB/c mice, and $5~10{\times}10^4$ of ameba trophozoites were intranasally inoculated for infection after splenectomy and/or immunization. ELISA technique was applied for the detection of seum IgG antibody levels. Experimental animals were divided into 4 groups; I. splenectomized and immuniEed; ll. splenectomized only; III. immunized only; IV. not splenectomized nor immunized. The results obtained were as follows: 1. Mortality rates of splenectomized and immunized mice in group I (38.1%) and immunized only in group III (25.0%) were lower than those of not immunized mice in group II (50%) and control group, IV (46.4%). 2. Survival times of mice in group I, II, III and IV were $20.1{\pm}3.6$, $17.3{\pm}4.5$, $20.4{\pm}7.0$ and $19.6{\pm}7.6$ days respectively, and there were no significant differences between them. 3. ELISA values (absorbance at 492nm) of group I (1, $10{\pm}0.29$) and group III ($1.31{\pm}0.28$) were significantly higher than that of group IV($0.24{\pm}0.37$) at day 31 of infection (p<0.05). Conclusively, it is presumed that humoral immunity against N. fowleri may operate as ever, after immunization, even though the mouse was splenectomized.
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