• 한국어병학회지
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• 제6권2호
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• pp.205-218
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• 1993

On the development of hirame(Paratichtys olivaceus) culture, outbreak of scuticociliata infection was reported to cause severe damage in Japan. To establish effective measures for isolation and cultivation of this ciliate, we tried to culture this pathogenic ciliate using medium for bacteria and fish cell lines in vitro. Scuticociliata from the brain tissues of infected fish was aseptically inoculated to CHSE-214 cells cultured in MEM-10 without antibiotic. Scuticociliata grew well and the number of ciliate reached $10^6\;cells/ml$ at temperatures of $15^{\circ}C$ to $20^{\circ}C$ for 10d. The number of ciliate cultured in the cell lines is 10 times higher than the numbers cultured in the liquid medium alone. This ciliata could be cloned by dilution method. Scuticociliata isolated could grow well on 42 different cell lines that were established from marine fish, warm freshwater fish, and salmonids. This ciliate could be preserved in liquid nitrogen for more than 6 months. Subsequently, we observed the optimal temperature and salinity for growth, and tested the sensitivities of this organism to formaldehyde, flagyl(Metronidazole), Ekuteshin(Combination compound of sulfamonometoxin and ormethoprim), and ozonixation. Optimal temperature for growth was $25^{\circ}C$ and salinity was 1.0 to 1.5%. Washed scuticociliata was killed by formaldehyde at the concentration of 50ppm for 10min, but was not completely killed even at a high concentration of 400ppm for 20min in MEM-5. Flagyl and Ekuteshin can inhibit the growth of scuticociliata at the concentration of 1,000 and $100{\mu}g/ml$ in MEM-10, respectively. More than 99% of this scuticociliata could be killed by ozonization at a dose equivalent to $1.0mg/\ell$ oxidant for 30sec in sea water. Isolated scuticociliata showed the pathogenicity to the cultured hirame by artificial infection(I. P. injection, $10^5\;cells$/fish). The number of scuticociliata in the water could be counted by most probable number(MPN) method using tissue culture, and the minimum detectable number was $1.8\;cells/\ell$. The number in the reservoir tank for water supply to the culture tank was 110 cells/l. After cleaning by elimination of the sediments from of the reservoir tank and disinfected with formaldehyde, number of scuticociliata decreased and was counted less than $1.8\;cells/\ell$ and infection rate of cultured hirame was decreased.

• 한국어병학회지
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• 제10권2호
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• pp.177-186
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• 1997

양식넙치에서 분리한 스쿠티카섬모충을 배양배지(P2Y1-DW배지)를 사용하여 순수분리한 후 배양온도에 의한 성장시험, 6종류 배양배지에서의 성장시험, 약제감수성시험을 하였다. 배양온도시험에서는 $25^{\circ}C$에서 $1.50{\times}10^3$충체/ml로 가장 좋았고, $15^{\circ}C$가 $1.03{\times}10^2$ 충체/ml로 가장 낮았다. 6종류의 배양배지에서의 성장시험에서는 3일째에 P1Y1-S배지가 $2.6{\times}10^4$ 충체/ml로 가장 좋았으며 DW배지에서가 $2.0{\times}10^2$ 충체/ml로 가장 낮았다. 5일째에는 배양배지 모두 감소하는 경향을 나타내었다. 스쿠티카섬모충의 분열방법은 앞부분을 밑으로 부착한 후 형태가 점점 서양배모양으로 변하고 시간이 경과함에 따라 2개의 세포로 분열하여 유영하다가 성충크기인 $40{\mu}m$로 되는데 부착개시부터 성충까지 소요되는 시간은 대략 3시간 전후였다.

• 한국어병학회지
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• 제19권3호
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• pp.189-195
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• 2006

A subtracted cDNA library of a marine scuticociliate, Philasterides dicentrarchii, in response to 17β-estradiol exposure was constructed using suppression subtractive hybridization (SSH). As a result of SSH, 275 clones were isolated, and among them, only glutathione peroxidase (GPX) gene was isolated as an antioxidative enzyme responding to 17β-estradiol. The semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis revealed that the transcription of GPX gene of P. dicentrarchii was clearly increased by exposure to 17β-estradiol. The GPX transcription was also clearly increased by exposure to xenoestrogens such as bisphenol A (BPA) and genistein.