• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.88-89
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• 2013

A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

• The Journal of Fisheries Business Administration
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• v.34 no.2
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• pp.75-90
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• 2003

When we think of fundamental problems of the aquaculture industry, there are several strict conditions, and consequently the aquaculture industry is forced to change. Fish aquaculture has a structural supply surplus in production, aggravation of fishing grounds, stagnant low price due to recent recession, and drastic change of distribution circumstances. It is requested for us to initiate discussion on such issue as “how fish aquaculture establishes its status in the coastal fishery\ulcorner, will fish aquaculture grow in the future\ulcorner, and if so “how it will be restructured\ulcorner” The above issues can be observed in the mariculture of yellow tail, sea scallop and eel. But there have not been studied concerning seabream even though the production is over 30% of the total production of fish aquaculture in resent and it occupied an important status in the fish aquaculture. The objectives of this study is to forecast the future movement of sea bream aquaculture. The first goal of the study is to contribute to managerial and economic studies on the aquaculture industry. The second goal is to identify the factors influencing the competition between production areas and to identify the mechanisms involved. This study will examine the competitive power in individual producing area, its behavior, and its compulsory factors based on case study. Producing areas will be categorized according to following parameters : distance to market and availability of transportation, natural environment, the time of formation of producing areas (leaderㆍfollower), major production items, scale of business and producing areas, degree of organization in production and sales. As a factor in shaping the production area of sea bream aquaculture, natural conditions especially the water temperature is very important. Sea bream shows more active feeding and faster growth in areas located where the water temperature does not go below 13∼14$^{\circ}C$ during the winter. Also fish aquaculture is constrained by the transporting distance. Aquacultured yellowtail is a mass-produced and a mass-distributed item. It is sold a unit of cage and transported by ship. On the other hand, sea bream is sold in small amount in markets and transported by truck; so, the transportation cost is higher than yellow tail. Aquacultured sea bream has different product characteristics due to transport distance. We need to study live fish and fresh fish markets separately. Live fish was the original product form of aquacultured sea bream. Transportation of live fish has more constraints than the transportation of fresh fish. Death rate and distance are highly correlated. In addition, loading capacity of live fish is less than fresh fish. In the case of a 10 ton truck, live fish can only be loaded up to 1.5 tons. But, fresh fish which can be placed in a box can be loaded up to 5 to 6 tons. Because of this characteristics, live fish requires closer location to consumption area than fresh fish. In the consumption markets, the size of fresh fish is mainly 0.8 to 2kg.Live fish usually goes through auction, and quality is graded. Main purchaser comes from many small-sized restaurants, so a relatively small farmer and distributer can sell it. Aquacultured sea bream has been transacted as a fresh fish in GMS ,since 1993 when the price plummeted. Economies of scale works in case of fresh fish. The characteristics of fresh fish is as follows : As a large scale demander, General Merchandise Stores are the main purchasers of sea bream and the size of the fish is around 1.3kg. It mainly goes through negotiation. Aquacultured sea bream has been established as a representative food in General Merchandise Stores. GMS require stable and mass supply, consistent size, and low price. And Distribution of fresh fish is undertook by the large scale distributers, which can satisfy requirements of GMS. The market share in Tokyo Central Wholesale Market shows Mie Pref. is dominating in live fish. And Ehime Pref. is dominating in fresh fish. Ehime Pref. showed remarkable growth in 1990s. At present, the dealings of live fish is decreasing. However, the dealings of fresh fish is increasing in Tokyo Central Wholesale Market. The price of live fish is decreasing more than one of fresh fish. Even though Ehime Pref. has an ideal natural environment for sea bream aquaculture, its entry into sea bream aquaculture was late, because it was located at a further distance to consumers than the competing producing areas. However, Ehime Pref. became the number one producing areas through the sales of fresh fish in the 1990s. The production volume is almost 3 times the production volume of Mie Pref. which is the number two production area. More conversion from yellow tail aquaculture to sea bream aquaculture is taking place in Ehime Pref., because Kagosima Pref. has a better natural environment for yellow tail aquaculture. Transportation is worse than Mie Pref., but this region as a far-flung producing area makes up by increasing the business scale. Ehime Pref. increases the market share for fresh fish by creating demand from GMS. Ehime Pref. has developed market strategies such as a quick return at a small profit, a stable and mass supply and standardization in size. Ehime Pref. increases the market power by the capital of a large scale commission agent. Secondly Mie Pref. is close to markets and composed of small scale farmers. Mie Pref. switched to sea bream aquaculture early, because of the price decrease in aquacultured yellou tail and natural environmental problems. Mie Pref. had not changed until 1993 when the price of the sea bream plummeted. Because it had better natural environment and transportation. Mie Pref. has a suitable water temperature range required for sea bream aquaculture. However, the price of live sea bream continued to decline due to excessive production and economic recession. As a consequence, small scale farmers are faced with a market price below the average production cost in 1993. In such kind of situation, the small-sized and inefficient manager in Mie Pref. was obliged to withdraw from sea bream aquaculture. Kumamoto Pref. is located further from market sites and has an unsuitable nature environmental condition required for sea bream aquaculture. Although Kumamoto Pref. is trying to convert to the puffer fish aquaculture which requires different rearing techniques, aquaculture technique for puffer fish is not established yet.