Spermatogenesis and the reproductive cycle in male Spisula sachalinensis were investigated by cytological and histological observations. The morphology of the spermatozoon has a primitive type and is similar to those of other bivalves in that it contains a short midpiece with four mitochondria surrounding the centrioles. But spermatozoon of this species has not axial rod and satellite body in the midpiece. The morphologies of the sperm nucleus type and the acrosome shape of this species have a globe-shape type and modified cap-like shape, respectively. The spermatozoon is approximately 40-45 μm in length including the sperm nucleus length (about 1.35 μm), acrosome length (about 1.50 μm) and tail flagellum. The axoneme of the sperm tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. The axoneme of the sperm tail shows a 9+2 structure. The spawning period of these species lasts from June to July, and the main spawning occurs in July when seawater temperatures are greater than 20°C. The male reproductive cycle of this species can be categorized into five successive stages: early active stage (October to January), late active stage (February to April), ripe stage (April to June), partially spawned stage (June and July), and spent/inactive stage (August to September).
GDNA was isolated from the jedo venus clam (Protothaca jedoensis, Lischke) from Boryeong (jedo venus clam from Boryeong JVCB) and Wonsan (jedo venus clam from Wonsan; JVCW) located in the West Sea and the East Sea of Korean Peninsula, respectively and we performed clustering analyses, DNA polymorphisms and the populations genetic variations. In the present study, the seven decamer primer generated the one hundred and eleven major/minor specific bands in JVCB population and ninety four-specific bands in JVCW population. Seven primers generated the unique shared bands to each population of one hundred and seventy-six, on average of 25.1, in JVCB population from Boryeong and three hundred thirty, on average of 47.1, in JVCW population from Wonsan, respectively. The dendrogram obtained by the seven oligonucleotides primers, indicates two genetic clusters. Especially, two Protothaca between the individual WONSAN no. 12 and BORYEONG no. 10 showed the longest genetic distance (0.537) in comparison with other individuals used. Accordingly, RAPD analysis showed that the JVCB was a little more genetically diverse than the JVCW population. This result implies the genetic similarity owing to rearing in the same and/or similar circumstances or inbreeding within the JVCW population. So to speak, JVCB population may have high levels of genomic DNA variability owing to the introduction of the wild individuals from the other sites to sampling sites although it may be the geographically diverse distribution of this species. However, it was confirmed that it did not appear like that really in this study. We feel convinced that RAPD analysis discovered a significant genetic distance between two Protothaca population pairs (P<0.001). The existence of population discrimination and genetic diversity between two Protothaca populations was identified by RAPD analysis.
In order to estimate the necessity of refrigerated storage of fresh seafood for short-term storage, and evaluate the effect of refrigerated storage on pen shell Atrina pectinata and clam Ruditapes philippinarum collected from Jang-su of Deukryang Bay and I-mok of Sunchen Bay in South Korea, the counts of coliform, Escherichia coli and total aerobic bacteria in A. pectinata and R. philippinarum under non-refrigerated (28±1℃) and refrigerated storage conditions (4±1℃) were determined. The results indicated that the storage at temperature of 4℃ possessed significant effects on inhibiting bacterial growth in live seafood. And refrigerated storage had different effect on A. pectinata and R. philippinarum. Different species and culture environments significantly influenced the initial and ultima bacteria counts. This study confirmed that refrigerated storage for short-term storage of live seafood was necessary, and indicated that the effect of refrigerated storage was influenced by comprehensive effectors.
Ultastructural studies of oocyte degeneration and follicle cells in female Spisula sachalinensis are described for clams collected from Jumunjin, Gangwondo, Korea. The follicle cells playan integral role in vitellogenesis and oocyte degeneration by assimilating products originating from the degenerated oocytes (thus allowed the transfer of yolk precursors needed for vitellogenesis). The functions of the follicle cells include phagocytosis and intracellular digestion of products originating from oocyte degeneration. During the period of oocyte degeneration, follicle cells of this species probably have lysosomal systems for the breakdown and reabsorption of various phagosomes(phagolysosomes) in the cytoplasm for nutrient storage; this process has been observed in other bivalves.
소라 (Batillus cornutus) 외투막의 조직화학적 특성 및 미세구조를 광학현미경과 투과전자현미경을 이용하여 기재하였다. 소라의 외투막은 결합조직층을 단층상 피층이 위아래로 둘러싸고 있는 구조였으며, 상피층은 내장 기관계 쪽을 향하고 있는 내부상피층과 패각 쪽의 외부상피층으로 구분된다. 상피층에 의해 둘러싸인 결합조직층은 주로 교원섬유와 근섬유다발로 구성되며, 혈림프동이 발달하고 있었다. 외투막의 각정부 상피층에 분포하는 점액세포들은 산성 및 중성 점액다당류를 함유하고 있었으며, 중간부와 가장자리의 상피층에 분포하는 점액세포들은 비황화 강산성의 당단백질 점액성분을 함유하는 것으로 나타났다. 외투막의 두께, 상피층의 두께, 혈림프동의 면적은 각정부에서 가장자리로 갈수록 낮아지는 경향을 보였다. 투과전자현미경으로 관찰하였을 때 외투막의 상피층은 단층으로 원주형 상피세포, 섬모세포, 흡수세포, 분비세포로 구성되어 있었다. 외투막 상피세포들은 원주형으로 장방형의 핵을 가지며, 자유면에는 미세융모들이 발달되어 있었다. 상피세포 사이에는 폐쇄띠, 폐쇄대와 수지상막구조로 연결되어 있었다. 섬모세포는 자유면에 섬모와 미세융모를 가지며, 세포질의 상부에는 다수의 미토콘드리아를 가지고 있었다. 상피층에서는 흡수기능을 가지는 두 종류의 세포가 관찰되었다. 이들은 원주형으로 미세융모, 음소포, 미토콘드리아 그리고 전자밀도가 다양한 용해소체들을 함유하고 있었다. 소라 외투막에서는 네 종류 (A, B, C, D)의 분비세포를 구분할 수 있었는데, 이들은 모두 단세포선으로 확인되어 대부분의 복족류와 이매패류의 외투막에서 보고된 선세포들과 유사한 특징을 보였다.
The histochemical characteristics and ultrastructure of the mantle in the spiny top shell, Batillus cornutus were described using light and electron microscopy. The simple epidermal layer wrapped on the top and bottom of the centrally located connective tissue. And then the epidermal layer were divided into the outer epidermal layer near a shell and the inner epidermal layer closed to the visceral mass. The connective tissue layer was composed of the collagen fiber muscularfiber bundle and hemolymph sinus. Mucous cells in the apical mantle contained acid and neutral mucopolysaccaride, and acidic carboxylated mucopolysaccaride in the mid and marginal mantle. The mantle thickness, epidermal layer thickness and hemolymph sinus area displayed a trend of reduction from the marginal zone to the apical zone. From TEM observation, it was possible to distinguish epithelium, ciliated cell, absorptive cell and secretory cell in the epidermal layer. The epithelia were columnar and the nucleus was elliptical. The free surface were covered with microvilli. The lateral membranes of epithelium was connected with neighboring cells by the zonular occludens, zonular adherens and membrane interdigitation. Ciliated cell on free surface had cilia and microvilli, and numerous mitochondria in the apical cytoplasm. In the epidermal layer, it observed 2 type cells having absorptive function. The absorptive cells were columnar in shape, and contained microvilli, pinocytotic vesicles, mitochondria and lysosomes of various electron density. Secretory cells can be divided into four types (A, B, C, D) depending on the cell shape and characteristics of secretory granules. These cells were unicellular glands and had similar characteristics to previously reported on the mantle of the gastropod and bivalves.
With the purpose to estimate the possibility of short-term storage and cryopreservation for sperm of Charonia sauliae, which is a potential preparation for its artificial reproduction and further research, in this study, protocols for short-term storage and cryopreservation of trumpet shell sperm was optimized. The effects of different immobilizing solutions, dilution ratios were estimated for short-term storage. And the effects of different cryoprotectant extenders and freezing rates were estimated for cryopreservation in terms of motility and survival of sperm. The results indicated that the artificial sea water of 350 mOsmol/kg is a better immobilizing solution and sperm which was diluted at a ratio of 1:1 (v/v) had higher motility and survival rate during short-term storage. The effect of 5% dimethyl sulfoxide was significantly better than those of other cryoprotectant extenders. And a freezing rate of -20℃ min-1 showed better effect than other freezing rates. In conclusion, this study optimized some key factors of the short-term and cryopreservation of C. sauliae sperm, which can provide valuable data for germplasm conservation and artificial propagation of C. sauliae.
본 연구는 피조개의 용존산소에 대한 내성을 구명하기 위하여 경남 진해만에 서식하는 피조개를 대상으로 수온과 염분에 대한 용존산소 농도별 생존율, 산소소비율 및 외부 형태 변화 등을 조사하였다. 피조개의 용존산소에 대한 <TEX>$LC_{50}$</TEX>은 수온 <TEX>$15^{\circ}C$</TEX>-일반해수와 염분 <TEX>$26{\pm}1$</TEX> psu에서 7일 동안 각각 1.98mg/L (신뢰한계: 1.45-2.39 mg/L)와 2.63 mg/L (신뢰한계: 1.76-3.37mg/L)였다. 한편 <TEX>$25^{\circ}C$</TEX>-일반해수와 염분 <TEX>$26{\pm}1$</TEX> psu에서는 각각 3.89 mg/L (신뢰한계: 3.36-4.55 mg/L)과 3.74 mg/L (신뢰한계 3.19-4.43 mg/L)으로 고수온에서 용존산소에 대한 내성이 약한 것으로 나타났다. 산소소비율은 모든 실험구에서 저산소 노출기간동안 감소하였으며, 회복가능성을 파악하기 위하여 일반해수로 전환시킨 경우 산소소비율을 증가시키며 산소부채에 의한 회복반응을 보였으나 모두 폐사하였다. 저산소 노출기간동안 패각 각정부위의 색깔이 검게 변화하는 것이 관찰되었다. 이 자료는 피조개 집단 폐사원인 구명을 위한 기초자료로 활용할 수 있을 것으로 여겨진다.
The ark shell, Scapharca broughtonii were collected from the Jinhae bay in Kyungnam, Korea from April 2006 to October 2007. To assess how much they have tolerance against hypoxia under the compound condition of temperature and salinity, survivorship, oxygen consumption rate and external features of Scapharca broughtonii were measured. The 7 days-LC50 of dissolved oxygen (DO) for Scapharca broughtonii at 15℃-ordinary sea water and 26±1 psu was 1.98 mg/L (confidence limit: 1.45-2.39 mg/L) and 2.63 mg/L (confidence limit: 1.76-3.37 mg/L) respectively. Whereas 3 days-LC50 at 25℃-ordinary sea water was 3.89 mg/L (confidence limit: 3.36-4.55 mg/L) and 4 days-LC50 at 25℃-26±1 psu was 3.74 mg/L (confidence limit: 3.19-4.43 mg/L). Oxygen consumption rate with each experimental group was decreased during the period exposed to hypoxia. To recovery, they were replaced to ordinary and oxygen consumption rate was increased a little, but all died. The color of the umbo of shell in Scapharca broughtonii was changed dark during hypoxia. These data will provide important fundamental information for examining the causes of mass mortality of shellfish in the summer.
백합과 5종에 속하는 유용패류인 살조개, 바지락, 백합, 개조개 및 가무락조개의 체성분에 따른 유사 종간의 근육과 내장에 따른 일반성분 조성을 분석하였다. 분석 결과 근육과 내장의 일반성분의 경우 수분, 조단백질 및 탄수화물은 근육부위에서 내장부에 비해 전체적으로 높은 값을 나타났다. 이러한 결과는 조지방과 탄수화물을 제외한 다른 항목에서는 유의한 차이를 나타내지 않았다. 조지방의 경우 백합 내장에서 <TEX>$3.67{\pm}0.57%$</TEX>로 높게 나타났으며, 바지락 근육에서 가장 낮은 <TEX>$2.50{\pm}1.40%$</TEX>를 나타내었다. 탄수화물의 경우 백합의 내장에서 <TEX>$3.61{\pm}0.45%$</TEX> 나타났으며, 바지락과 살조개 내장에서 <TEX>$3.45{\pm}1.4%$</TEX> 및 <TEX>$3.39{\pm}0.07%$</TEX>로 높게 함유하고 있었다.
This study investigated the proximate composition in the muscle and viscera of five commercially valuable Korean Veneridae, Protothaca jedoensis, Ruditapes philippinarum, Saxidomus purprratus, Cyclina sinensis and Meretrix lusoria. The proximate analysis revealed that the moisture, crude protein and carbohydrate of muscle in higher than viscera. As results, except for the crude lipid and carbohydrate there was little significant differences among the moisture, protein and crude ash. The crude lipid contents was highest in the viscera of Meretrix lusoria, while lowest in the muscle of Ruditapes philippinarum. The carbohydrate content of viscera in the Meretrix lusoria, Ruditapes philippinarum and Protothaca jedoensis were 3.61±0.45%, 3.45±1.4% and 3.39±0.07%, respectively.
동양달팽이의 metallothionein 유전자는 염기서열 195개로 이루어져 있으며 65개의 아미노산으로 이루어져 있었다. 연체동물의 metallothionein 서열의 공식인 C-x-C-x (3) -C-T-G-x (3) -C-x-C-x (3) -C-x-C-K에 맞춰본 결과 알려진 공식과 일치하는 것을 확인할 수 있었으며 아미노산의 조성도 시스테인 (Cys) 이 30% 이상 함유하는 사실을 확인 할 수 있었다. BLAST 결과를 토대로 선정된 72개의 참고 서열 중 아미노산 레벨에서 가장 높은 스코어로 align 되는 서열은 Helix pomatia의 CD-MT 서열이었다. Clustalx를 통해 multiple align 한 후 Neighbor-Joining method 방법에 따라 phylodendrogram을 그려본 결과 Helix pomatia, Helix aspersa, Arianta arbustorum, Megathura crenulata 등과 같은 복족류 육산패들과 같은 그룹으로 묶여지는 것을 확인 할 수 있었다. Psipred 소프트웨어를 통해 2D 구조를 비교 분석 한 결과도 multiplealignment 및 phylodendrogram와 밀접한 관계가 있음을 알 수 있었다. 이러한 결과를 통해 EST를 통해 밝혀진 동양달팽이의 MT서열은 근연종들과 매우 일치함을 알 수 있었으며 MT 서열이 분류에 사용 될 수 있음을 확인시켜 주었다.
Numerous morphological studies on N. samarangae have been well conducted over the last ten years. In this context, we have attemtped to do molecular phylogenetic analysis by using metallothionein (MT) gene from N. samarangae. To this end, we cloned the full length cDNA of MT from cDNA library of N. samarangae. The complete cDNA sequences were obtained from the expressed sequence tag (EST) sequencing project of N. samarangae. The coding region of 195 bp gives an amino acid sequence of 65 residues including methionine. There are 5' (61 bp) and 3' (48 bp) untranslated region at both ends of the Ns-MT cDNA sequence. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of Ns-MT cDNA indicate that N. samarangae has similarity to land snails such as Helix pomatia, Helix aspersa and Arianta arbustorum.
