• Research in Plant Disease
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• v.12 no.3
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• pp.197-204
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• 2006

Crown gall on lower stem and root of chrysanthemum(Dendranthema grandiflorum Kitamura) was observed at Hwasung and Gumi in 2001 and 2004, respectively. Tumors were semi-round with rough surface texture of dark brown color. Nine isolates inducing gall formation on lower stem of chrysanthemum among twenty isolates from the tumor tissues were characterized. Their colonies were convex, glistening, circular with an entire edge and whitish or tannish cream in color on potato dextrose agar supplemented with 0.5% $CaCO_3$. The virulent isolates were rod-shaped with peritrichous flagellae, gram negative, aerobic and growing on D1M agar. Among nine virulent isolates, one isolate was identified as Agrobacterium rubi and eight isolates were A. tumefaciens based on biochemical and physiological characteristics, fatty acid profiles and substrate utilization patterns. A. tumefaciens had strong pathogenicity and broad host range compared with A. rubi. This is the first report on crown gall of chrysanthemum in Korea. To our knowledge, crown gall of chrysanthemum caused by A. rubi is first report in this study worldwide.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.23-30
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• 1996

대략 36,000 base pairs (bp)의 두 가닥짜리 DNA를 지놈으로 가진 사람 아데노바이러스 (Ad)는 DNA 상동성(相同性) 및 생물학적/생화학적 성격이 특이한 49개의 혈청형이 알려져 있는데, 이들 대부분의 Ad가 영유아군 및 면역능이 저하된 성인에서 치사적 결과를 초래할 수 있다. Ad의 세포향성(向性)(tropism)은 매우 다양하여 종류에 따라 상기도 감염, 각결막염, 영유아 장염등을 유발하는데 최근 Ad의 다양한 병원성에 대한 원인을 분자생물학적 수준에서 규명하려는 노력의 일환으로 지역에 따라 주되게 출현하는 Ad형 규명이 활발히 이루어지고 있다. Ad 동정/확인은 표면을 이루고 있는 group 공통항원인 hexon 단백질을 탐지하는 효소면역 측정법 (EIA)에 의하며, Ad형별은 Ad fiber의 세포독성 중화시험에의 한다. 그러나, 세포독성 중화시험이 엄청난 노동력 및 시간을 요구하면서도 민감도/특이도가 만족스럽지 못하여 이를 개선하기 위하여 검체 또는 세포배양에서 Ad DNA를 추출하여 제한효소 절단형태를 비교하는 방법이 개발되었는데 이는 세포배양에 잘 자라지 않는 바이러스주의 형별뿐만 아니라 지역 분리주들의 지놈 변형주를 관찰하는 분자생물학적/분자역학적 연구에도 도움이 되고 있다. 국내에는 Ad와 관련된 소아장염의 빈도가 rotavirus에 의한 것 다음으로 빈번한데도 Ad40/41외에 주되게 출현하는 장내 Ad형들이 전혀 규명된 바 없고, 한국형 Ad들의 지놈형태가 전혀 보고된 바가 없다. 또한 세계적으로 Ad형별 조사지역이 늘어감에 따라 유아장염과 연관된 Ad 역시 Ad40, 41이 외의 형들이 Ad40, 41을 능가하는 것으로 보고되고 있는 지역도 있으나 국내에서는 Ad40, 41이외의 형들은 그 역학적 중요도가 전혀 알려져 있지 않다. 이로서 본 연구의 목적은 Ad주들에 특이 중화항체를 이용한 세포독성 중화시험과 Ad DNA 절단법을 적용하여 한국형 장내 Ad주들의 형별을 처음으로 시도함과 동시에 1989-1991사이 출현한 Ad들의 유전적 변형을 관찰하려는 것이었다. 두 방법 모두 사용하였을 때 주되게 출현하는 장내 Ad형들은 Ad4l, Ad2, Ad7, Ad5, 및 Ad40이었다. Ad40/41-양성 검체를 제외한 Ad hexon-EIA양성들의 77.5%를 형별 할 수 있었던 Ad DNA의 제한효소 절단방법은 형들간의 교차중화로 특이성이 낮았던 중화방법 (47.5%)보다 매우 효율적이어서 두 가지 방법을 함께 적응하였을 때는 40주중의 81.5%인 35주를 형별 할 수 있었다. 또한Ad DNA 제한 효소 절단방법은 Ad7 변이주 (Ad7b)도 탐지 할 수 있었다.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.77-90
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• 1996

Nucleocapsid protein (NP)which exists in the particle of hantavirus and surrounds the viral RNA genome is one of the major structural proteins and plays role of antigen to elicit the antibody detected predorminantly right after infection of the virus in the patients of hemorragic fever with renal syndrome (HFRS)or experimental animals. NP is important target antigen in serological diagnostic system of HFRS utilizing whole antigens from the native virus particle, such as IFA, ELISA and Western blotting. Therefore, the preparation of this protein in the level of higher quantity and purity is desirasble for developed dianosis of the disease. The purpose of this study is the cloning of NP gene which exists in the S genome segment of Maaji (MAA) virus and expression of the gene to obtain qualified, genetically engineered NP to be utilized as an immunodiagnostic antigen. First of all, for the purpose of amplifing the MAA-NP gene by PCR, the specific primers were built from the known nucleotide sequence of Hantaan viral NP gene. The viral cDNA of the NP gene was synthesized by using the primers and RNase $H^-$ AMV reverse transcriptase. Thereafter, using this cDNA as a template, the NP gene was amplified specifically by Taq DNA polymrerase. The pT7blue (R)T-overhang vector systems were used for cloning of the amplified NP gene. The expression system was consisted of BL21 (DE3)pLysS and pET16b as a host and a plasmid repectively. Into Ndel site of pET16b, NP gene was ligated with cohesive end for the expression. Insertion of NP gene in the plasmid was confirmed by PCR and mini prep methods. For expression, IPTG was used and the expressed protein was characterized by Western blotting. The MAA-NP was expressed as the form of inclusion body (insoluble fraction)and the protein purified by affinity and metal chealating columns reacted specifically with the sera from patients of HFRS as to be tested by ELISA and Western blotting.

• Parasites, Hosts and Diseases
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• v.22 no.2
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• pp.259-266
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• 1984

Present study was undertaken to elucidate the changes of the ultrastructure of Naegleria fowleri trophozoite in brain tissue of mice and culture medium. Naegleria fowleri, 0359 strain, which used in this study was cultured in axonic liquid medium, CGVS medium. Each mouse was inoculated with amoebas intranasally under secobarbital anesthesia, and sacrificed on 7th day after the infection. Comparative observation of the ultrastructure of the amoebas in axonic culture and experimentally infected mice brain was done with transmission electron microscope. The results are summarized as follows: 1. The amoebas in mouse brain tissue were round in outline, whereas those of amoebas from axonic culture showed irregular appearance. 2. Mitochondria in the amoebas from axonic culture was oval, round and cylindrical shape and darkly stained, whereas those of the amoebas from mouse brain tissue showed dumbbell shape together with above forms. The stain was not unique, but light and/or dark. 3. Rough endoplasmic reticulum of amoebas in brain tissue was tubular, but from culture it was vesicular or tubular in shape. 4. Emity vacuoles were demonstrated in amoebas from culture, while food vacuoles with myelinated structures were abundant in those from tissue, suggesting a strong phagocytic activity. 5. Mouse brain tissue in ected were extensively destroyed, and Polymorphonuclear leukocytes were infiltrated predominantly with inflammatory lesion. Amoebas were observed in the vicinity of the capillary.

• Parasites, Hosts and Diseases
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• v.22 no.2
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• pp.229-237
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• 1984

The migration and distribution pattern of spargana in mouse body was observed after experimental infection through mouth. The spargana were obtained from the snake, Natris tigrina lateralis, caught in Hoengseong-gun, Kangwon-do. A total of 28 male mice (ICR strain), 21∼259 in body weight, were fed each with 5 scolices (and necks) of spargana and killed after 10 minutes to 14 days. Systemic autopsy was performed on each mouse to recover the spargana. The results are as follows: 1. The spargana were found to penetrate into the stomach or duodenal wall of mice as early as 10 minutes after infection. They completed the penetration within 30 minutes and appeared in abdominal cavity. It was observed that spargana did not migrate tangentially along the gut wall but directly perforated the wall. 2. After 1 hour to 1 day the majority of spargana distributed in abdominal cavity of mice except a few which migrated to muscles or subcutaneous tissues. 3. It was within 7 days that nearly all of the spargana migrated to subcutaneous tissues. Out of total 28 in number found from subcutaneous tissues, 13 distributed around neck region, 12 around trunk and other 3 on head of mice and the most common sites were submandibular and subscapular areas. There was nearly no host tissue reaction to migrating spargana. 4. The initial length of spargana given was 4 mm in average but it increased to 12 mm after 7 days and to 35 mm after 14 days. The results suggest that spargana orally given to mice penetrate the gut wall within 30 minutes followed by escaping into abdominal cavity, and after passing through thoracic cavity or abdominal wall they anally Localize in subcutaneous tissues chieay around neck region within 7 days.

• Parasites, Hosts and Diseases
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• v.29 no.2
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• pp.121-128
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• 1991

The degree of attraction of Toxoplasma gondii to vertebrate cells varies with cell type and cell phase. Human promyelocytic leukemia cells, HL-60, were synchronized by double thymidine block method and co-cultured with Toxoplasma for 1 hr at each cell stage to investigate the cell cycle specific susceptibility of parasites to host cells. For 30 hr the average number of Texoplasma that invaded was a little changed except at 3 hr from G1/S phase boundary which concurred with the peak point of DNA synthesis. At 3 hr which is a relatively short interval compared to whole S phase, modification of cells by parasitic invasion was most remarkable. The number of Toxoplasma that penetrated was increased to more than sin times. The shape of the cells became sludgy and almost indiscernible by strong accessibility of parasites only for an hour of mfd-S phase. The same auctuation was also observed at the second peak of S phase but weakly. This suggests that there be surface molecules concerning with the attachment of Texoplasma to the host cells, which is expressed at special point of S phase. further studies on the specific protein or similar molecules related could be carried out using synchronized HL-60 cells.

• Parasites, Hosts and Diseases
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• v.29 no.2
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• pp.161-172
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• 1991

The complete life cycle of Echinostoma hertense has been maintained in the laboratory, using Lymnaea persia snails and Rana nigromaculata tadpoles as the first and second intermediate hosts. ICR mice was used as the definitive host. Within the egg of 5. hotense, the miracidium was fully matured in 13 days of incubation at $29~30^{\circ}C$. The miracidium was $93.8{\times}53.6{\;}{\mu\textrm{m}}$ in average size, covered with numerous cilia of $7~11{\;}{\mu\textrm{m}}$ length. The epidermal plates were arranged in 6-8-4-2 formula. The first generation rediae ($1.19{\times}0.27{\;}mm$ in average size) were observed in 14 days after miracidial challenge to the snails, and the second generation rediae ($1.40{\times}0.26{\;}mm$ in average size) in 30 days. The average sixte of the cercaria was $295.5{\times}145.0{\;}{\mu\textrm{m}}$. Their head crown was poorly developed, and collar spines were not yet observed. After a cercarial challenge to the tadpoles, all of the tadpoles became infected and the average worm recovery rate was 88.5%. The majority of the metacercariae (75.5%) were recovered from the muscle of the tadpole's posterior body and the rest (24.3%) from their gills. The metacercariae from the tadpoles were elliptical, and $167.7{\times}129.9{\;}{\mu\textrm{m}}$ in average size. The recovery rate of adults from the mice was difFerent by the age of the metacercariae grown in the tadpoles. The metacercariae younger than 5 hrs could not infect mice whereas those older than 6 hrs could infect mice. The recovery rate became higher as the metacercaria matured, with the peak recovery rate of 90.0 % at the metacercarial age of 9 days. Thereafter the recovery rate decreased to 55.0% at the age of 50 days. As shown by the above results, the whole life cycle of E. hcrtense has been completed in the laboratory. At least 55~58 days were required to maintain one egg-to-egg cycle of E. hortense.

• Parasites, Hosts and Diseases
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• v.35 no.2
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• pp.79-86
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• 1997

Toxoplasmn gonnii, an intracellular protozoan infecting many kinds of eukaryotic cells, has been used to experimentally infect macrophages, epithelial cells, fibroblasts, and various cancer cells, but rarely T and B Iymphocytes or granulocytes. The present study was performed to determine the susceptibility of murine (BALB/c or CBA) splenic T and B llrnphocytes, and granulocytes to infection trio T. gondii RH tachyzoites. The ultrastructure of the infected host cells was observed by TEM, and the degree of intracellular parasite proliferation was quantified using 3H-uracil uptake assay. At 24 hrs post-culture, the host cell cytoplasm was found to contain 1 or 2, or a maximum of 7-8 tachyzoites. Infected T Iymphocytes demonstrated a peripherally displaced nucleus, a parasitophorous vacuole enveloping the parasite, and an increased number of mitochondria. In B Iymphocytes infected with tachyzoites, RER was not well developed compared to uninfected B Iymphocytes. Uninfected granulocytes contained many electron dense granules, but T. gondii-infected granulocytes demonstrated a decreased number of granules. Based on the 3H-uracil uptake assay. the susceptibility of T and B Iymphocytes, and granulocytes, to infection with T. gonnii tachyzoites was fairly high irrespective of cell type and strain of mouse. This strongly suggests deterioration in the functioning of infected host immune cells.

• Parasites, Hosts and Diseases
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• v.35 no.4
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• pp.233-238
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• 1997

A small coastal village of Muan-gun, Chollanam-do, was surveyed for intesti- nal fluke infections, especially heterophyids such as Heterophwes nocens and Ftsiniopsis summc by fecal examination on 108 inhabitants. The egg Positive rate of heterophyids was very high. 75.0%, and that of other parasites was comparatively low.0.9-3.7% by parasite species . After treatment of 20 patients showing high E. P. G. with praziquantel and purging with Mgs04, total 3,864 specimens of H. nocens were collected from the diarrheic stools of all the patients treated (3-1,338 individuallyl and total 703 p. summc were harvested from 18 patients (1-170 individually), together loth several other species of flukes. Other flukes included Stictodora jlscotn (164 specimens from 4 patients), Heterophwopsis continuo (2 from 2 patients), and Gwmnophalloides seoi (4 from 3 patients). From this study, the sutra- veyed coastal area of Muan-gun, Chollanam-do was proven to be a new endemic focus of H. nocens and p. summa. The occurrence of a few infected cases surf:mists that this area should also be a low-grade endemic area of S. Juscctc, H. continuo, and G. seoi.

• Parasites, Hosts and Diseases
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• v.35 no.4
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• pp.259-264
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• 1997

This study was carried out to determine whether dermal mast cell responses to Parasoninn westemoni in an abnormal host, the mouse, were dependent on the site of metacercarial inoculation. In mice during subcutaneous infection, the number of der- mal mast cells were increased significantly (p<0.05) at the first week ($38.3/\textrm{mm}^2$) and then persisted at a high level until the sixth week ($45.2/\textrm{mm}^2$) of infection compared with PBS- injected (control) mice (range: $19.4-25.1/\textrm{mm}^2$). In mice during oral infection, the number of dermal mast cells were increased significantly (p<0.05) at two weeks ($33.5/\textrm{mm}^2$) after infection and remained at these levels thereafter compared loth non-infected (control) mice (range: $17.4-22.3/\textrm{mm}^2$). In mice both during subcutaneous and oral infection, the recruited dermal mast cells showed extensive degranulation at the second week (68.4%) and 60.7%, respectivelyl, reached a peak at the third week (81.4%, and 92.1%, respectively) and then declined slightly thereafter. By contrast, in both control mice, about 10% of dermal mast cells were degranulated. In conclusion, this study suggests that dermal mast cell responses to p. westemcni in mice are dependent on cutaneous sensitization by larval excretory-secretory antigens, irrespective of infection route.