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In-situ hybridization 법을 사용한 양식 넙치, Paralichthys olivaceus의 바이러스 감염 질병 특성 고찰
도정완 ( Jeong Wan Do ),이남실 ( Nam Sil Lee ),정승희 ( Sung Hee Jung ),김경길 ( Kyung Kil Kim ),최혜승 ( Hye Sung Choi ),박정우 ( Jeong Woo Park ),김이청 ( Yi Cheong Kim ) 한국어병학회 2013 한국어병학회지 Vol.26 No.3
PCR (polymerase chain reaction) 법은 신속하고 정확하여 바이러스성 질병진단을 위해 널리 사용되지만 조직병리학적인 정보를 제공하지 못한다. 반면에 in-situ hybridization (ISH) 법을 사용하면 바이러스를 빠르게 검출할 수 있을 뿐 아니라 조직에서의 분포도알 수 있다. 본 연구에서는 RSIV, VHSV, 그리고 VNN바이러스들의 조직내 분포 및 조직 병리학적인 특성을 확인하기 위하여 이 바이러스들에 감염된 에 감염된 양식 넙치의 어류의 다양한 조직들을 대상으로 ISH 법을 적용하였다. 그 결과 이들 세 종류의 바이러스가 각각 다른 조직 및 세포들에 감염함을 확인 할 수 있었다. 본 연구 결과는 ISH법이 어류 병원성 바이러스의 신속 검출 뿐 아니라 조직 병리학적인 특성 확인에도 유용함을 제시한다. Polymerase chain reaction (PCR) is the most rapid and widely used method to detect viral pathogens. However, this method does not provide histopathologic nature of the virus. In situ hybridization (ISH) with oligonucleotide probes is attractive because it is a rapid method for detection and identification of viral pathogens at sites of tissue infection. In order to understand the histopathologic characterictics of Red sea bream iridovirus (RSIV), viral-hemorrhagic septicemia (VHS) virus and viral nervous necrosis (VNN) virus to cultured olive flounder, we her applied ISH method to various kinds of olive flounder tissues with PCR-positive for these three viruses. We found that these viruses showed different tissue tropism and were detected from different cell types. Our results suggest that ISH is useful not only in rapid detection of viral pathogens but also in understanding the histopathologic characters of specific viral pathogens.
감성돔(Acanthopagrus schlegelii) 치어의 포르말린에 대한 내성 및 조직학적 반응
명정인 ( Jeong In Myeong ),민병화 ( Byunng Hwa Min ),박미선 ( Mi Seon Park ),황형규 ( Hyung Kyu Hwang ),도정완 ( Jeung Wan Do ),정경일 ( Kyung Il Jeoung ),장영진 ( Young Jin Chang ),정달상 ( Dal Sang Jeong ) 한국수산과학회 2013 한국수산과학회지 Vol.46 No.6
Black seabream juveniles Acanthopagrus schlegelii held at 20°C were exposed to formalin at concentrations of 50 to 400 ppm, and tolerance values were determined by calculating median lethal concentration values (LC50) through probit analysis. The 12-, 24,48, 72- and 96-h LC50 values for formalin were 297, 233, 171, 162 and 157 ppm, respectively. The histological effects of formalin on gill and liver tissues in this fish were determined. No histological effects were observed in the control group. The intensity of cell damage increased with the concentration of, and duration of exposure to, formalin. Hyperplasia, separation and epithelial necrosis,epithelial lifting, lamellar synechiae and collapsed lamellae were observed in gill tissues exposed to formalin. Hepatic lesions in liver tissues of fishes exposed to formalin were characterized by cloudy swelling of hepatocytes, necrosis, cytoplasmic vacuolization,deposition of pigments, spongiosis hepatis, nuclear hypertrophy, dilation of sinusoids and bile stagnation. The LC50 values and histological results obtained in this study will aid in designing treatment regimens to minimize toxic side effects and increase efficacy.
양식동남아산 뱀장어(Anguilla bicolor pacifica)의 Heterosporis anguillarum 감염
김진도 ( Jin-do Kim ),도정완 ( Jeong-wan Do ),최혜승 ( Hye-sung Choi ),조혜인 ( Hyae-in Jo ),이남실 ( Nam-sil Lee ),김영대 ( Young-dae Kim ) 한국환경생물학회 2014 환경생물 : 환경생물학회지 Vol.32 No.4
Shortfin eel (Anguilla bicolor pacifica) is a species of commercial importance and its production is greatly affected due to the infection by Heterosporis anguillarum. In this study, we evaluated the effect of H. anguillarum infection on the growth of Shortfin eel. A disease that trunk muscle of cultured shortfin eel, Anguilla bicolor pacifica, were irregular and resulted in death, breakout of the commercial eel culture farm. We observed that the trunk muscle of infected eels were irregular and represented white or yellowish externally. Histopathologically, a great numbers of large or small spores and sporophorocysts were also observed in degenerated muscle layer. The cloning of specific gene of H. anguillarum, encoding small subunit ribosomal RNA (SSU-rRNA)was amplified by the polymerase chain reaction(PCR) from the muscle lesion of diseased eel. The size of clone gene is well matched with the size of small subunit ribosomal RNA of H. anguillarum and thus confirming the infection by H. anguillarum.
Aeromonas veronii의 실험적 감염에 의한 양식 메기, Silurus asotus의 병리학적 변화
김진도 ( Jin Do Kim ),도정완 ( Jeong Wan Do ),최혜승 ( Hye Sung Choi ),서정수 ( Jung Soo Seo ),정승희 ( Sung Hee Jung ),조혜인 ( Hyae In Jo ),박명애 ( Myung Ae Park ),이남실 ( Nam Sil Lee ),박성우 ( Sung Woo Park ) 한국환경생물학회 2013 환경생물 : 환경생물학회지 Vol.31 No.4
Recently, a new disease showing symptoms such as epidermal exfoliation and muscular necrosis occurred in cultured Korean catfish. Although the mortality of fishes was low but the economic damages owing to loss of commercial value were severe. The authors isolated the causative agent from diseased fish and observed pathological changes both in naturally and artificially infected fish. The causative bacteria was identified as Aeromonas veronii. Subsequently we observed the daily death and pathological symptoms of artificially infected fish with Aeromonas veronii. Symptoms of artificially infected fish were similar to those of naturally infected fish and all fish died within 7 days after infection. Histopathological changes on the naturally infected fish revealed severe congestion and necrotic degeneration in the liver, spleen and kidney. Some bacterial aggregates with inflammatory degeneration were observed in the heart, and congestion and fibrosis in the lamina propria of digestive tube were predominant. In artificially infected fish, skin erosion and necrotic degeneration of muscle tissue around injected region were particularly manifested. Degeneration of hepatocytes in liver and hyalic degeneration around ellipsoids in spleen were partially observed. However, there were no predominant signs in digestive tube in artificially infected fish.
해상가두리양식장에서 사육한 교잡종 (참돔♀ × 감성돔♂), 긴키돔, 참돔 및 감성돔의 질병 특성
최혜승 ( Hye Sung Choi ),도정완 ( Jeong Wan Do ),박명애 ( Myoung Ae Park ),안철민 ( Chul Min Ahn ) 한국어병학회 2012 한국어병학회지 Vol.25 No.3
교잡종, 참돔 및 긴키돔은 수온이 19.7℃에서 24.0℃로 상승하는 7월부터 8월에 가장 높은 성장률을 나타낸 반면, 감성돔은 수온이 23.0℃에서 19.9℃로 하강하는 9∼10월에 가장 높은 성장률을 나타내었다. 생존율은 조사기간 동안 교잡종이 가장 높은 것으로 조사되었지만 이러한 높은 생존율이 잡종 강세의 특성에 의한 것인지에 다른 요인에 의한 결과인지는 추후 자세한 연구가 필요할 것으로 생각된다. 감성돔 종 0∼90%, 참돔 20∼100%, 감성돔 10% (10월), 긴키돔 20∼100%로 유의적인 차이는 없었지만 교잡종에서 다소 낮은 편이었다. Hybrid of red seabream (Pagrus major) and Japanese red seabream (Pagrus major) grew rapidly from to August (WT 19.7℃∼24.0℃) while black seabream (Acanthopagrus major) did from September to October (WT 19.9∼23. 0℃). Survival rate of hybrids (85.5%) was a significantly higher than those of the other fish species (20%). Green liver syndrome were observed in black sea bream and Japanease red seabream but did not in hybrids. Microcotyle tai infected 0∼90% in hybrid, 20∼100% in red seabream, 10% in black sea bream and 20∼100% in Japanese red seabream but no significant difference was found among each fish species.
Amprolium hydrochloride가 넙치 Paralichthys olivaceus의 약물대사 유전자 발현에 미치는 영향
박상협 ( Sang Hyup Park ),김창환 ( Chang Han Kim ),도정완 ( Jeong-wan Do ),최혜승 ( Hye-sung Choi ),김이경 ( Yi Kyung Kim ) 한국어병학회 2023 한국어병학회지 Vol.36 No.2
본 연구에서는 넙치의 해독 과정에서 amprolium hydrochloride의 영향을 평가하기 위해 수행되었다. 이전 연구에서 보고된 amprolium의 LD<sub>50</sub> 값을 이용하여 두 가지 실험을 진행하였다. 첫 번째는 30마리의 넙치를 5개의 대조군 및 실험군으로 나누었고 4, 8, 16, 32 mg/kg 용량의 amprolium을 근육 내 주사 투여하였다. 주사 후 8, 24, 48 시간에 간과 신장을 적출하여 약물 대사 효소와 전염증성 사이토카인 유전자의 발현을 분석하였다. 32 mg/kg 용량의 실험군에서 IL-1β mRNA의 높은 발현을 확인하였고, CYP1A는 이와 반대의 결과를 보였으며, 간에서 UGT와 GST mRNA의 발현은 유의하게 감소하는 것을 확인하였다. 또한 신장에서 amprolium 주사 투여 후 약물 대사 효소와 사이토카인 유전자의 억제가 관찰되었다. 또 다른 실험에서는 4, 8, 16, 32 mg/kg과 60, 80, 100, 120 mg/kg의 용량을 설정하여 근육 내 주사 투여하였다. 주사를 완료하고 6일 후 간을 적출하여 유전자의 발현을 확인하였다. IL-1β의 발현은 4 mg/kg 용량 실험군에서 유의적으로 매우 높은 발현을 보였다. GST의 mRNA 발현 또한 4 mg/kg 용량 실험군에서 높은 발현을 보였다. 결론적으로 우리의 결과는 amprolium이 가축 산업의 가장 안전한 합성 항콕시듐 약물 중 하나로 간주되지만 넙치의 간접 또는 직접적인 물리적 또는 생물학적 독성을 유발하는 것으로 판단된다. This study was undertaken to evaluate the effect of amprolium hydrochloride on detoxification process of olive flounder Paralichthys olivaceus. A series of two experiment was performed based on the LD<sub>50</sub> value obtained for amprolium. First, thirty flounder (average weight 230.27 g; average length 27.99 cm) was randomly allocated into five groups. Treatment was carried out using intramuscular injection of amprolium at the dose levels of 4, 8, 16, and 32 mg/kg body weight. At 8, 24 and 48 h post injection, liver and kidney were collected for expression assay of drug metabolizing enzymes and pro-inflammatory cytokine genes. We found that the interleukin-1β (IL-1β) mRNA level were induced at 32 mg/kg and CYP1A genes showed the opposite pattern, while UDP-glucuronosyl-transferase (UGT1A7) and GST were significantly reduced in the liver. Moreover, the suppression of drug metabolizing enzymes and cytokine gene in the kidney was observed after treatment. Another treatment was carried out using intramuscular injection with 4, 8, 16, and 32 mg/kg and 60, 80, 100, 120 mg/kg body weight. At 6 days post injection, liver was collected. The IL-1β expression was markedly induced in the experimental group treated with 4 mg/kg. In addition, glutathione S-transferase (GST) mRNA level was higher in the group with 4 mg/kg. In conclusion, our data suggests that amprolium seem to cause direct or indirect physical, or biological toxicity of flounders, although this drug is considered one of the safest synthetic anticoccidial drugs of the livestock industry.