• 한국양식학회지
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• 제10권3호
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• pp.227-237
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• 1997

Five species of intertidal clams including Ruditapes philippinarum, Tegillarca granosa, Solen strictus, Heteromacoma irus, and Coecella chinensis were tested for the presence of the protozoan parasite, Perkinsus sp. using fluid thioglycollate medium (FTM) fortified with antibiotics and histological techniques. Each individual clam was placed in a test tube filled with 10ml FTM, placed in totally dark place, and incubated over a week. After incubation the clam tissues were stained with Lugol's iodine solution and examined under a light microscope to find out any hypnospores of Perkensus sp. in the tissues. Cross-sections of the clams were also embedded in paraffin, sliced to 3um, and stained with Harry's hematoxylene and Picro eosine to observe the presence of tomont or trophozoites. Perkinsus sp. were found in the presence of tomont or trophozoites. Perkinsus sp. were found in the tissues of R. philippinarum collected from Kangjin and Wando, along the south coast of Korea. However, Perkinsus sp. was not found in four other species of clams nor R. philippinaurm collected from Kimnyong and Waido in Cheju. A size-dependent Perkinsus sp. infection was found in R. philippinarum collected rom Kangjin and Wando the clams smaller than 15mm in shell width do not exhibit and Perkinsus sp. while other clams greater than 20mm in shell width exhibit almost 100% infection. To determine the number of Perkinsus sp. in the clams, FTM cultured clam tissues were digested with 2M NaOH solution and the number of hypnospores in the tube were counted. The number of hypnospores counted from the tissues indicated that each Manila clam contains 100,000 to 3,500,000 Perkinsus cells or 20,000 to 1,000,000 cells per gram tissue wet weight. The results of cell counts also suggests that such a high occurrence of Perkinsus sp. in the clam may cause mortality, as already reported from other studies of Perkinsus spp.

• 한국패류학회지
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• 제29권1호
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• pp.23-32
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• 2013

The purpose of this study is to evaluate the effects of oil dispersant and ultraviolet (UV) radiation on the toxicity of crude oil. The toxicity of water-accommodated fraction (WAF) and chemically enhanced water-accommodated fraction (CEWAF) of Hebei Spirit crude oil was investigated in the embryo of the Manila clam, Ruditapes philippinarum with- and without ultraviolet radiation. The WAF and CEWAF with- and without UV radiation affected significantly the embryonic development of R. philippinarum. The EC50s of WAF without UV, WAF with UV, CEWAF without UV, and CEWAF with UV were 2.82, 0.79, 1.60, and 0.45 g/L, respectively. CEWAF was 1.6 times more toxic than WAF. UV radiation increased crude oil toxicity to 3.6 times for both WAF and CEWAF. The oil dispersant and UV radiation did not affect the acute toxicity to the embryo but retarded the period of embryonic development up to 26%. R. philippinarum proved to be a sensitive species to reflect the toxic effects of oil spill combined with oil dispersant and UV radiation. It is suggested that the chemical analyses on the WAF and CEWAF is important for the identification and quantitative explanation of the phototoxic compounds in crude oil.

• Ocean and Polar Research
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• 제32권3호
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• pp.247-254
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• 2010

Single cell PCR analysis and light and scanning electron microscopic techniques were utilized to identify free living bivalve larvae in the coastal waters of Tae-an, on the west coast of Korea. Through DNA sequencing, venerid clam larvae were isolated and identified as Ruditapes philippinarum (99% similarity) and Meretrix lusoria (99%). Under microscopic observation, the D-veliger stage of R. philippinarum exhibited symmetrical shoulder angles and an elliptical ventral form. In contrast, M. lusoria displayed asymmetrical shoulder angles and a round ventral form in the umbonal stage. Size of the R. philippinarum larvae was $156{\pm}22{\mu}m$ in length, $126{\pm}12{\mu}m$ in height, $92{\pm}14{\mu}m$ in width with a length: height ratio of 1.23. Meretrix lusoria was $202{\pm}44{\mu}m$ in length, $161{\pm}35{\mu}m$ in height, $96{\pm}38{\mu}m$ in width with a length: height ratio of 1.25. Experimental results indicate that morphological and molecular characteristics provide evidence for the larval identification of these two venerid clam larvae species in nature.

• 한국발생생물학회지:발생과생식
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• 제12권1호
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• pp.41-49
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• 2008

한국 곰소만산 암컷 바지락(Ruditapes philippinarum)의 난모세포의 발달 및 퇴화과정 중 일어나는 미세구조적 변화에 관해 기술하였다. 난소소낭은 영양성분을 저장하는 포상결체조직세포(VCT cell)들의 기질에 의해 둘러싸여 있다. 난 형성과정 초기에 초기난황형성난모세포와 보조세포(follicle cell)들 사이에 desmosome-like gap junction들이 나타났다. 난황형성과정은 골지체, 미토톤드리아, 조면소포체가 결합되어 작용하는 자율합성과정을 통하여 일어나며, 난황막 형성에 앞서 난황형성과정의 초기와 난모세포 퇴화에 없어서는 안될 역할을 하는 것으로 나타났다: 보조세포들은 식세포작용 및 난모세포 퇴화로부터 유래되는 산물들의 세포내소화에 관여한다. 이들의 기능은 난황형성에 필요한 난황전구체들로 전환이 일어나게 한다. 또한, 보조세포들은 난황형성난모세포들이 이용하는 글리코겐과 지방적들을 세포질 내에 영양물질로 축적, 저장할 수 있다. 보조세포들은 난모세포가 퇴화되는 기간 중에는 난모세포의 붕괴를 위해 리소조옴 체제를 가지며, 영양물질 저장을 위해 세포질 내에 존재하는 여러 종류의 파고소옴(phagosome)들을 흡수하였다. 그러나 보조세포들은 난황형성과정에서 난황형성을 위한 난황전구체의 주된 공급원이 아니라고 추정된다.

• 한국패류학회지
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• 제25권2호
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• pp.135-143
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• 2009

바지락 (Ruditapes philippinarum) 은 우리나라 조간대에 널리 분포하며 잘 발달된 생체조절 능력을 가지고 있어 환경모니터링 종으로서 가치가 높은 생물이다. 본 연구는 산업활동으로 해양환경에서 그 농도가 높아지고 있으며 생물체에 강한 독성을 가진 카드뮴이 바지락의 세포성 면역에 미치는 영향을 연구함으로써 바지락의 면역 기작을 이해하고 환경 모니터링 기법으로의 적용 가능성을 조사하기 위하여 실시되었다. 실험을 위하여 바지락은 $50\;{\mu}g/l$의 카드뮴에 8일간 노출되었으며, 유세포 분석기를 이용하여 혈구의 미분화 세포의 구성비율, DNA 손상도, 혈구괴사율, 혈구 apoptosis 비율 및 혈구사망율을 측정하였다. 조사 결과 카드뮴에 노출된 바지락은 대조구와 비교하여 미분화세포, DNA 손상도, 세포괴사율, 세 apoptosis 비율 및 세포 사망율이 통계적으로 유의하게 증가하였다. 특히 apoptosis 비율은 세포괴사율보다 높게 증가함에 따라 바지락 혈구의 경우 본 연구에서 사용된 카드뮴의 농도는 세포괴사보다는 apoptosis를 유도하는데 더 효과적인 농도인 것으로 판단된다. 본 연구를 통하여 중금속 오염에 대한 바지락의 세포성 면역반응은 유세포 분석기를 이용하여 정량이 가능하였으며, 세포성 면역반응을 이용한 환경모니터링에 유용하게 사용될 수 있을 것으로 기대된다.

• 한국패류학회지
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• 제28권2호
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• pp.145-156
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• 2012

이 연구에서는 바지락 (Ruditapes philippinarum) 의 번식량 및 유생발생을 관찰하였으며, 이를 위하여 수정란 및 유생을 21일 간 실내사육수조를 이용하여 배양하였다. 바지락은 2011년 8월 제주시 김녕에서 채집 실험 수조로 옮겨졌으며, 다음날 자연적인 방란 방정이 관측되었다. 바지락 수정란은 19시간의 배 발생과정을 거쳐 21일 후 포복피면자유생(pediveliger) 으로 발달하였다. 조직학적 방법을 이용하여 바지락의 생식소를 관찰한 결과, 방란, 방정 전 모든 개체는 성적으로 완숙한 상태였고, 산란 후 모든 개체에서 부분산란이 관찰되었다. 효소면역학적 방법 (ELISA) 에 의한 번식량 측정 결과, 암컷 바지락의 생식소 지수는 (GSI) 산란 전 28.6%에서 산란 후 17.3%로 11.3% 감소하였으며, 추정된 포란수는 6,998,658 eggs였다. 이 연구서 광학현미경 및 전자현미경을 이용하여 관찰된 바지락 유생 발달 및 번식량은 향후 바지락 인공종묘 산업 및 바지락 자원관리에 있어 중요한 자료로 활용될 것으로 사료된다.

• 한국해양생명과학회지
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• 제3권2호
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• pp.59-66
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• 2018

갯벌의 건강 수준에 대한 평가는 갯벌에 서식하는 생물의 건강에 의해 평가될 수 있다. 생체지표유전자의 발현 분석을 통하여 갯벌에 서식하는 생물의 건강 수준을 평가할 수 있다. 본 연구의 목적은 heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), glutathione S-transferases (GST) 및 thioredoxin (TRX)과 같은 생체지표유전자를 이용하여 서해안 갯벌의 건강을 평가하는 것이다. 이들 유전자는 스트레스, 면역 및 항산화 관련한 유전자들로서 이들 유전자의 발현을 통해 생물의 건강 정도를 관찰하는 데 사용할 수 있다. 본 연구에서는 서해안의 8개 정점에서 바지락(Ruditapes philippinarum)을 수집했다. 유전자의 발현은 RT-qPCR 방법으로 분석하였다. 연구 결과 Hsp70, Hsp90, GST 및 TRX 유전자들의 발현이 8개 정점에서 차별적으로 발현되는 것으로 나타났다. 특히, Hsp90 및 GST의 발현 또는 Hsp70 및 TRX의 발현은 유사하였다. 이것은 각 유전자에 특이적으로 반응하는 물질이 존재하는 것으로 생각된다. 따라서 이화학적 분석결과에 근거하여 분석에 적합한 유전자를 선택할 수 있다고 생각한다. 이 결과는 Hsp70, Hsp90, GST 및 TRX 유전자는 갯벌의 건강을 평가하기 위한 생체지표유전자로서의 역할을 수행함을 시사한다.

• Fisheries and Aquatic Sciences
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• 제4권4호
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• pp.208-218
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• 2001

We have investigated the gonad index (GI), gonadal development, reproductive cycle, first sexual maturity, sex ratio, the number of spawned eggs and spawning frequency of the Manila clam, Ruditapes philippinarum. Samples were collected from the intertidal zone of Komso Bay, Korea from January to December in 1999. Monthly changes in the gonad index (GI) and condition index showed a similar pattern in the reproductive cycle. The spawning period was once a year between early June and early October, there was a spawning peak between July and August when seawater temperature was over $20^{\circ}C$. The reproductive cycle of this species can be categorized into five successive stages; early active (February to March), late active (April to May), ripe (April to August), partially spawned (June to October), and spent/inactive stage (August to March). Percentages of first sexual maturity of female and male clams of l5.1-20.0mm in shell length were $56.3\%$ and $60.0\%$, respectively, and $100\%$ for the clams >25. mm. The sex ratio of individuals >15.1 mm in shell length was about 1:1 $(\chi^2= 0.02,\;p>0.05)$. Number of the eggs released from each clam by the induction increased as the size of clam in terms of shell length increased. Mean number of the eggs from the second induction of the spawning was $75.35-84.30\%$ $(average\;79.81\%)$ of the number of the eggs released in the first spawning. Our data indicated that R. philippinarum in Komso Bay has one major spawning peak with over two minor spawning, and the interval of each spawning was estimated to be approximately 15-17 (average 16.5) days.

• 한국수산과학회지
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• 제49권6호
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• pp.800-806
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• 2016

In total, 151 Escherichia coli isolates from Ruditapes philippinarum in Gomso Bay were analyzed for their susceptibility to 18 different antimicrobial agents and for genes associated with virulence. For virulence genes, each strain of the isolates was positive for the enterotoxigenic E. coli (ETEC)-specific heat-stable toxin (estA), enteroinvasive E. coli (EIEC)-specific invasion-associated locus (iaa) gene and enteropathogenic E. coli (EPEC)-specific attaching and effacing (eae) gene. According to a disk diffusion susceptibility test, resistance to ampicillin was most prevalent (23.2%), followed by resistance to amoxicillin (22.5%), ticarcillin (20.5%), tetracycline (18.5%), nalidixic acid (12.6%), ciprofloxacin (10.6%), streptomycin (9.9%), and chloramphenicol (6.6%). More than 35.8% of the isolates were resistant to at least one antimicrobial agent, and 19.9% were resistant to four or more classes of antimicrobials; these were consequently defined as multidrug resistant. Minimum inhibitory concentration (MIC) ranges for the antimicrobial resistance of the 15 different antimicrobial agents of 54 E. coli strains were confirmed by varying the concentrations from $32-2,048{\mu}g/mL$. Overall, these results not only provide novel insights into the necessity for seawater and R. philippinarum sanitation in Gomso Bay but they also help to reduce the risk of contamination by antimicrobial-resistant bacteria.

• 한국패류학회지
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• 제26권3호
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• pp.245-254
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• 2010

For the studies of germ cell development and maturation in the ovary, the gametogenic cycle and the number of spawning seasons per year in female Ruditapes philippinarum were investigated by quantitative statistical analysis using an Image Analyzer System. Compared with the results by qualitative and quantitative analyses, monthly variations in female gonad indice by qualitative histological analysis showed a pattern similar to that of the female gametogenic cycle calculated by quantitative statistical analysis. The number of spawning seasons occurred once per year, from June to October. In quantitative statistical analysis using an image analyzer system, monthly changes in the portions (%) of the ovary area to total tissue areas in females increased in March and reached a maximum in May, and then showed a rapid decrease from June to October when spawning occurred. And also monthly changes in portions (%) of follicle areas to the ovary area and in portions of oocyte areas to ovarian tissue areas in females began to increase in March and reached a maximum in May, and then. rapidly dropped from June to October when spawnig occurred. From these data, it is apparent that the number of spawning seasons occurred once per year, from June to October. Monthly changes in the number of the oocyte per $mm^2$ and in mean diameter of the oocyte in captured image which were calculated for each female slide showed a maximum in May and reached the minimum from December to February. Therefore, female R. philippinarum showed a unimodal gametogenic cycle during the year.