• Journal of Aquaculture
• /
• v.10 no.3
• /
• pp.227-237
• /
• 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.

• The Korean Journal of Malacology
• /
• v.29 no.1
• /
• pp.23-32
• /
• 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
• /
• v.32 no.3
• /
• pp.247-254
• /
• 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.

• Development and Reproduction
• /
• v.12 no.1
• /
• pp.41-49
• /
• 2008

Ultrastructural changes occurring during the course of development and degeneration of oocytes in female Ruditapes philippinarum (Adams & Reeve, 1850) are described for clams collected from Gomso Bay, Korea. During the early stages of oogenesis, desomosome-like gap junctions localized between the early vitellogenic oocyte and the follicle cells. Vitellogenesis occurs through a process of autosynthesis, involving the combined activity of the Golgi complex, mitochondria and rough endoplasmic reticulum, and heterosynthesis in which extraovarian precursors are incorporated into oocytes by endocytotic activity, involving the basal region of the early vitellogenic oocytes prior to the formation of the vitelline envelope. The follicle cells appear to play an integral role in vitellogenesis and oocyte degeneration: phagocytosis and intracellular digestion of products originating from oocyte degeneration. These functions can permit a transfer of yolk precursors necessary to vitellogenesis, and they can accumulate nutrients in the cytoplasm, as glycogen and lipids, which can be employed by the vitellogenic oocyte. During the period of oocyte degeneration, follicle cells may have lysosomal system for breakdown, and resorb various phagosomes in the cytoplasm for nutrient storage. But follicle cells probably are not the major source of yolk precursors in vitellogenesis.

• The Korean Journal of Malacology
• /
• v.25 no.2
• /
• pp.135-143
• /
• 2009

The Manila clam, Ruditapes philippinarum, has been considered as a sentinel species due to dominant distribution along the coast of Korea and well developed regulatory system. In order to develop and understand immune responses of the Manila clams, clams were exposed to $50\;{\mu}g/L$ of cadmium chloride (Cd) for 8 days and monitored the cellular immune parameters of the hemocytes including blast cell composition, DNA damage, necrosis, apoptosis and hemocyte mortality using a flow cytometer. The results showed that all immune parameters analyzed in the present study increased remarkably compared to the controls and the increases were statistically significant. Apoptosis rate was higher than necrosis rate in the clams exposed to Cd suggesting that apoptosis was preferably induced by the concentration of Cd used in the present study. Our study indicates that the measurement of cellular immune responses of the Manila clam using flow cytometer will be a useful technique for assessment of heavy metal contamination in marine environment.

• The Korean Journal of Malacology
• /
• v.28 no.2
• /
• pp.145-156
• /
• 2012

Larval development of the Manila clam Ruditapes philippinarum reared in an indoor tank system was examined in this study using light microscope and scanning electron microscope. To induce spawning and subsequent larval development, clams were collected from the intertidal zone at Gim-nyeong harbor in Jeju Island in August 2011. After 2 days of rearing in the tank, all Manila clams spawned in the midnight. Non-feeding trochophore larvae appeared 7hrsafter fertilization and the first D-shape larvae could be observed at 19 hrs. Twenty one days after fertilization the pediveliger larvae crawling on the bottom of the tank with well-developed foot were observed. Histology indicated that all the clams used in this study were in the ripe stage prior to spawning and the gonad-somatic index (GSI), a ratio of the egg mass to the tissue weight, of the ripe female measured by ELISA was 28.6%. The GSI of female clam declined to 17.3% after the massive spawning in the tank, suggesting that Manila clam discharged 40% of the total eggs during the first spawning event. In conclusion, spawning and subsequent larval development of Manila clam was successfully carried out in this study using an indoor tank system, and the information obtained in the present study could be useful in future Manila clam hatchery development.

• Journal of Marine Life Science
• /
• v.3 no.2
• /
• pp.59-66
• /
• 2018

The assessment of level of health of the tidal flats can be evaluate by health of organisms inhabit the tidal flats. It is possible to evaluate the precise health level of organisms inhabit the tidal flats using analysis of expression of biomarker genes. The purpose of this research is to evaluate the health of the tidal flats on the west coast using biomarker genes such as heat shock protein 70 (Hsp70), heat shock protein 90 (Hsp90), glutathione S-transferases (GST) and thioredoxin (TRX). These genes are stress, immune, and antioxidant related genes that can be used to look at the health of an organism through gene expression. In this study, we collected manila clam (Ruditapes philippinarum) in 8 analysis areas on the west coast. Expression of the genes was analyzed by RT-qPCR method. Results showed that, the expression of Hsp70, Hsp90, GST and TRX genes were differentially expressed in the 8 analysis areas. In particular, the expression of Hsp90 and GST or the expression of Hsp70 and TRX were similar. This means that there is a substance that reacts specifically to each gene. Therefore, I think suggest that the based on the results of physicochemical analysis, it can be selected genes suitable for analysis. These results suggest that Hsp70, Hsp90, GST and TRX were played roles in biomarker for assessment of the health of tidal flats.

• Fisheries and Aquatic Sciences
• /
• v.4 no.4
• /
• pp.208-218
• /
• 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.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.49 no.6
• /
• pp.800-806
• /
• 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.

• The Korean Journal of Malacology
• /
• v.26 no.3
• /
• pp.245-254
• /
• 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.