CONSTRUCTION OF THE DEEP-SEA MICROBIAL IN SITU INCUBATION SYSTEM TO SEARCH FOR DIVERSIFIED DEEP-SEA BAROPHILIC MICROORGANISMS

深海微生物原位培养系统按以下规范构建：最大使用深度：6000米最长系泊时间：1年重量和尺寸：尽可能小而轻，以便潜水器可以带到深海海底操作：电池和计时器取水：油管泵系统（Yamato, Co., Japan）8套无菌“血袋系统”（TERFLEX，日本），容量为150 ml /套在8套培养箱中，事先加入少量不同底物，从培养箱中取出的所有试管连接到泵管系统。计时器设定在船系泊到深海后一天开始运行。它的结构是由一个计时器控制的耐压电池室内的电池，迫使电机旋转，使切割器旋转，切断所有的进气管道，并使与孵化器相连的管泵系统将周围的水吸进孵化器。1996年11月21日，“白湖号”在日本海沟附近，在38 01′n，145 32′e的水深5000 m处，用多套不同深度的海流仪进行了深海微生物原位培养系统的停泊。1997年9月2日，它在被R/V Tansei-maru设置了大约9个月后，在巡航中被成功地回收到船上。整套培养系统是安全的。然而，遗憾的是，在8套油管泵系统中，只有一套运行良好，另外7套根本没有将任何深海样品装入孵育袋。在加入ZoBell 2216e作为底物的孵育袋中，观察到深海细菌密集生长（1.1x10^8个细胞/ml）。在1atm条件下的实验培养中，它们完全没有生长。相反，它们在2℃的压力条件下（500大气压）表现出良好的生长，目前正在进行这些菌株的分离和系统发育研究。少

The Deep-Sea Microbial in situ Incubation System was constructed with the following specificaton ;Maximum depth of use : 6,000 m Maximum duration of mooring : 1 year Weight and size : as small and light as possible so that submersible can bring to the deep-sea floor Operation : battery and timer Water sampling : tubing pump system (Yamato, Co., Japan) Incubators : 8 sets of sterile " Blood Bag System " (TERFLEX,Japan) with volume of 150 ml eachInto the 8 sets of incubators, small volume of the different substrates were added beforehand, and all the tubes from the incubators were connected to the tube pumping systems. The timer was set to start operation one day after the mooring to the deep-ocean. It was constructed that the battery inside the pressure tolerant battery house operated by a timer forces the motor to rotate which makes the cutter to rotate and cut off all the intake tubes to incubators and makes the tube pumping systems connected to the incubators to take ambient water in … More to the incubators.The Deep-Sea Microbial in situ Incubation System was moored at the site of the Japan Trench at a depth of 5,000 m on November 21,1996, at 38ﾟ01'N,145ﾟ32'E with several sets of water current meters at different depths during R/V Hakuho-maru cruise. On September 2,1997, it was successfully retrieved on board the vessel during cruise by R/V Tansei-maru after setting it for about 9 months. Whole set of incubation system was safe. Unfortunately, however, it was found that in the 8 sets of tubing pumping systems only one worked well and other 7 sets didn't take any deep-sea water sample at all into incubation bags. In one incubation bag in which ZoBell 2216 E was added as a substrate, dense growth of deep-sea bacteria (1.1x10^8 cells/ml) was observed. Experimental culture of them under 1 atm condition did not show any growth at all. On the contrary they showed good growth under pressurized condition (500 atm) at 2 C.Isolation of these bacterial strains and study for phylogeny of them are now undertaken. Less