• 대한수의학회지
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• 제10권2호
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• pp.21-51
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• 1970

Series of experiments were conducted to determine lethal does of X-ray and Neutron on Toxoplasma gondii. strain RH and IRI. As well morphological changes of Toxoplasma gondii irradiated or not were compared by use of electron microscope. The pathogenicity test of the irradiated and nonirradiated Toxoplasma gondii was made in mice guinea-pigs, rabbits and pigs: The letahl dose of X-ray and Neutron on RH and IRI strain and the growth rate between two strains after irradiation were shown little differences. Morphological changes were not observed until 18th passage was made. After then, the growth rate was decreased apparently, and atrophied forms were frequently observed in electron microscope. Survival time of animals inoculated with irradiated strain was longer than that of animals giving non-irradiated strain, and Toxoplasma gondii were isolated from all the dead animals. But it is of interest that pigs survived after injection of Toxoplasma gondii remained health and much attempts were failed toisolate Toxplasma gondii remained health and much attempts were slaughtered them. Animals were succumbed after injection of Toxoplasma gondii without any relationship with serum titers. (HA antibody).

• Parasites, Hosts and Diseases
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• 제59권6호
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• pp.565-572
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• 2021

Toxoplasma gondii ME49 infections are typically diagnosed by serological tests. However, serological diagnosis of RH strain-induced toxoplasmosis remains unknown. In order to develop seradiagnosis of above 2 kinds of infections, we generated recombinant virus-like particles (VLPs) displaying the T. gondii rhoptry protein 4 (ROP4) and evaluated their potential in T. gondii ME49 or RH strain infection diagnostics. Mice were orally infected with either the tachyzoites of T. gondii (RH) or cysts of T. gondii (ME49) at various dosages, and sera were collected at regular intervals. ELISA-based serological tests were performed to assess IgG, IgM, and IgA antibody responses against ROP4 VLP antigen and tissue lysate antigen (TLA). Compared to TLA, IgG, IgM, and IgA levels to ROP4 VLP antigen were significantly higher in the sera of T. gondii RH-infected mice 1 and 2 week post-infection (PI). T. gondii-specific IgG antibody was detected at 1, 2, 4, and 8 week PI in the T. gondii ME49-infected mice with infection dose-dependent manner. These results indicated that the ROP4 VLP antigen was highly sensitive antigens detecting T. gondii RH and ME49 antibodies at an early stage.

• 대한수의학회지
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• 제33권2호
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• pp.285-287
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• 1993

Toxoplasma gondii frozen by the method of very slow cooling ($-30^{\circ}C/3hrs.{\rightarrow}\;-60^{\circ}C/lhr.{\rightarrow}\;-196^{\circ}C$ of liquid nitrogen)were inoculated mice after storage in the liquid nitrogen for 5 years. The results obtained were as follows; 1. Toxoplasma gondii was detected in mice 4~5th day after intraperitoneal inoculation of organisms stored in the liquid nitrogen once a year for 5 years. 2. Toxoplasma gondii could be preserved longer than 5 years in liquid nitorgen being evidenced by the in vivo test.

• 대한수의학회지
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• 제13권1호
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• pp.67-73
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• 1973

Toxoplasma gondii (Tp), RH strain, was inoculated into cultured buffy coat cells obtained from the swine blood. The main reason for adopting swine lies in the animal's unusual susceptibility to Tp, As for the culture method used in the experiment, those well proved methods practised by Cho, Merchant, Moore and Tarnvik were mainly referred to as a starting point: hence, the author's method has been turned out to be the modified or supplementary form of those methods. Observations were made on the phase of multiplication of Tp in the cytoplasm. The results obtained were as follows: 1. Better growth and multiplication of Toxoplasma gondii were noticeably observed in the swine buffy coat cell, inoculated after three-to-five day cultivation of the cell. 2. In the lapse of the observation period, there appeard Toxoplasma gondii rarely available in the earlier stage, which had been inoculated into the cell after three-to-five day cultivation. In other words, Toxoplasma gondii started to show itself in seven or eight hours after inoculation, most outstandingly noticeable between twenty four hours and forty eight hours. Thereafter the disintegration stage of Toxoplasma gondii was observed.

• 대한수의학회지
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• 제39권6호
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• pp.1151-1160
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• 1999

This study was conducted to detect the toxoplasma-specific DNA in peripheral blood collected from cats experimentally infected with Toxoplasma gondii (RH strain) and from domiciled cats by B1 gene-base polymerise chain reaction(PCR). The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with parasite detection by mouse inoculation(MI). And also, latex agglutination test(LAT) and indirect fluorescent antibody test(IFAT) were conducted to detect the fluctuation of serum antibodies compared with the detection of toxoplasma by PCR and MI. Toxoplasma B1 gene PCR was shown consistently high sensitivity and the results obtained by PCR agreed completely with those from MI. All blood samples collected before infection with T gondii gave negative results by PCR and MI. Also, toxoplasma Bl gene PCR was not cross reaction with Neospora caninum DNA and normal cat leucocyte as controls. The toxoplasma-specific DNA was detected by PCR in blood of 5 cats experimentally infected with T gondii 6 days after infection and the detection of this specific-DNA was long lasted in blood for 64 days after infection. The detection of toxoplasma-specific DNA by PCR could be identified as few as 10 tachyzoites and the isolation of T gondii by MI could be isolated as few as 1 tachyzoite from tenfold serial dilution of T gondii with normal cat blood, respectively. In healthy domiciled cats, the toxoplasma-specific DNA and the parasite were detected and isolated in blood from 3 of 56(5.3%) cats by both PCR and MI, respectively. In the results of antibody test from the total 56 heads of healthy domiciled cats, the positive rates are 15(26.7%) by LAT and 19(33.9%) by IFAT. These results suggest that PCR detection of toxoplasma can be applied as a sensitive and specific diagnostic and research tool.

• Parasites, Hosts and Diseases
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• 제41권3호
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• pp.147-154
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• 2003

Toxoplasma gondii tachyzoites were isolated from the blood of an ocular patient, and have been successfully passaged in the laboratory, for over a year, by peritoneal inoculation in mice. The isolated parasite was designated the Korean Isolate-1 (KI-1) and its characteristics were compared with those of the RH strain, a well-known virulent strain originating from a child who suffered from encephalitis. The morphology, pathogenicity, infectivity and cell culture characteristics of the KI-1 were similar to those of the RH strain. Both RH and KI-1 antigens were detected by an anti-T gondii monoclonal antibody (mAb), Tg563, against the major surface protein SAG1 (30 kDa), whereas no reaction was observed against an anti-Neospora caninum mAb, 12B4. The KI-1 was confirmed as an isolate of T. gondii. A long-term laboratory maintenance and characterization of a local T gondii isolate is reported for the first time in the Republic of Korea.

• Parasites, Hosts and Diseases
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• 제48권3호
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• pp.195-201
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• 2010

We studied on the proteomic characteristics of Toxoplasma gondii KI-1 tachyzoites which were originally isolated from a Korean patient, and compared with those of the well-known virulent RH strain using 2-dimensional electrophoresis (2-DE), mass spectrometry, and quantitative real-time PCR. Two-dimensional separation of the total proteins isolated from KI-1 tachyzoites revealed up to 150 spots, of which 121 were consistent with those of RH tachyzoites. Of the remaining 29 spots, 14 showed greater than 5-fold difference in density between the KI-1 and RH tachyzoites at a pH of 5.0-8.0. Among the 14 spots, 5 from the KI-1 isolate and 7 from the RH strain were identified using MALDI-TOF mass spectrometry and database searches. The spots from the KI-1 tachyzoties were dense granule proteins (GRA 2,3,6, and 7), hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGRPTase), and uracil phosphoribosyltransferase (UPRTase). The spots from the RH strain were surface antigen 1 (SAG 1), L-lactate dehydrogenase (LDH), actin, chorismate synthase, peroximal catalase, hexokinase, bifunctional dihydrofolate reductase-thymidylate synthase (DHTR-TS), and nucleosidetriphosphatases (NTPases). Quantitative real-time PCR supported our mass spectrometric results by showing the elevated expression of the genes encoding GRA 2,3, and 6 and UPRTase in the KI-1 tachyzoites and those encoding GRA 7, SAG 1, NTPase, and chorismate synthase in the RH tachyzoites. These observations demonstrate that the protein compositions of KI-1 and RH tachyzoites are similar but differential protein expression is involved in virulence.

• Parasites, Hosts and Diseases
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• 제31권4호
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• pp.379-381
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• 1993

우태아혈청의 성분이 Toxoplasma의 숙주세포 침투를 억제하는 현상을 보여 이를 보고하고자 한다. MDCK세포를 숙주로 하여 Toxoplasma의 RH주를 첨가하였을 때, 우태아혈청 농도 1%(v/v) 에서 억제 효과가 나타나기 시작하여 5%일때 침투능을 반감시켰다. 우태아절청의 비동화 여부는 억제 효과에 영향을 미치지 못하였으며, 우태아혈청을 $95^{\circ}C$에서 10분간 처리하여 억제물질의 기능을 파괴시킬 때 억제 효과는 약간 감소되었다. Toxoprasma를 먼저 우태아혈청으로 처리한 후 숙주 세포에 첨가하였을 때 침투능에는 효과를 미치지 못하였으나, Tomplasma를 숙주세포와 배양하면서 우태아혈청을 첨가하였을 때에는 시간의 경과에 따라 침투능을 억제시켰다. 이상의 결과로 우태아혈청에 Toxoplasma의 숙주세포 침투를 억제하는 성분이 존재하는 것을 확인하였으며, 억제 방법이 침투하는 순간에 일어나며, 억제물질의 기능을 통해서라기 보다는 구조를 통해서 이루어진다고 추정할 수 있었다.

• Parasites, Hosts and Diseases
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• 제37권3호
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• pp.157-162
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• 1999

Serum from mouse orally ingested with tissue cyst forming stain (Me49) of Toxoplasma gondii was assayed by Western blot and immunofluorescene assay (IFA) to establish early responses in antigenicity of the parasite in mouse model of foodborne toxoplasmosis. Sera were collected weekly to blot the RH antigen transferred onto nitrocellulose paper after being separated by 12% SDS-PAGE. With the second week serum, 34 kDa protein (p34) was detected uniquely, and all antigens of T.gondii were detected with the sera from 3 or 4 weeks. p34 was not a member of the major surface membrane proteins and confirmed to be localized in the rhoptry by IFA. It was secreted into parasitophorous vacuolar membrane (PVM) during the entry into host cells. 10.3% of sera detected p34, while all the ELISA positive sera detected the band. It has diagnostic usefulness of presumed T.gondii infection. We suggest the name of the p34 protein as ROP9.

• Bulletin of the Korean Chemical Society
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• 제35권10호
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• pp.3071-3076
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• 2014

Proteomic analyses of the excretory/secretory proteins from the RH strain of Toxoplasma gondii have been performed to understand their functions in the host-parasite interaction. A total of 34 proteins were identified from LC/MS/MS analysis and their abundance was estimated by spectral counting methods. Among them, 8 species of micronemal proteins (MICs), 2 species of rhoptry proteins (ROPs), and 6 species of dense granular proteins (GRAs) were confirmed. Besides these, 18 species of protein were newly identified, and their cellular functions were estimated from sequence analysis. The three most abundant of the 34 identified extractor/secretory proteins-GRA1, GRA7 and GRA2-were confirmed to be highly expressed in T. gondii using the spectral count method. This phenomenon is another demonstration of the importance of GRA proteins for the penetration and survival of T. gondii.