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1 Feture Article Erozoru Kenkyu, 25, - Biosol Smpling Experiments t High Altitude Atmosphere nd Spce Akihiko YAMAGISHI Received November Accepted Jnury Abstrct Biosol microbesmpling experiments hve been conducted since the erly century. The reltive difficulty in smpling using ircrfts, blloons or rockets limited the number of smpling experiments. However, spore forming microbes nd Deinococci hve been smpled by the limited number of experiments, both re known to be extremely resistnt ginst extreme environments, such s UV nd desicction. Smpling devices re designed depending on the vehicle used for the smpling. The height dependence of the microbil density suggested the possible presence of microbes t the low erth orbit km. The microbe smpling experiment t ISS Interntionl Spce Sttionhs been proposed in Tnpopomission. We re going to use silic erogel for cpturing micro-prticles including biosol, if the mission is finlly ccepted. The device for the smpling will be uplifted by the vehicle HTV nd trnsferred to spce through irlock nd plced with robotic-rms. The device will be retrieved nd crried bck to the ground, where PCR nlysis will be conducted fter microscopic inspection of the biosol. Key WordsMicrobe Smpling Experiments, Spore Forming Microbes, Deinococci, UV-Resistnce, Desicction-Resistnce, Interntionl Spce Sttion, Tnpopo Mission. km km km Fig. 1 Fig. - - Deprtment of Moleculr Biology, Tokyo University of Phrmcy nd Life Science - Horinouchi, Hchioji-shi - Corresponding Author. E-mil : ymgish@toyku.c.jp Fig. 1 Schemtic representtion of the height dependence of temperture solid linend pressure broken line.

2 UV-A nmuv-b nmuv-c nm UV-A UV-B UV-C km UV-C Tble 1 km km km km km km Tble 1 Microbiologicl studies in the high tmosphere Reference Mission Smpling method Altitude km Microbil collection cultivted bcteri nd fungi Totl colony number Smpled ir volume ATP m Smpled ir volume STP m b Density ATP microorgnisms per m Density STP microorgnisms per m Rogers nd Blloon Meier, Sterile cylindricl smpling device descending by prchute Bcillus sp., Mcrosporium sp., Rhizopus sp., Penicillium sp. nd Aspergillus sp.... Soffen, Blloon Drwing ir through filter ppers by motor blower Penicillium sp..... Greene et Blloon l., ; Bruch, Drwing lrge volumes of ir by fn through polyurethne fom filter Predominnt isoltesmicrococci nd sporeforming rods, Aspergillus sp., Alternri sp., Penicillium sp. nd Cldosporium sp Fulton, Aircrft Air pss through geltin fom filter in smpling probe in front of bombrdier s comprtment... Predominnt isoltesbcilli, Micrococci, Aspergillus sp., Alternri sp. nd Hormodendron sp. c c c c c c Imshenetuky et l., Rocket Sterile medium exposed on the foreprt of the rocket during scent Mycobcterium sp., Micrococcus sp., Circinell sp., Aspergillus sp., Ppulspor sp. nd Penicillium sp. unknown unknown unknown unknown Yng et l., Aircrft Drwing ir through membrne filters by pumps. Deinococcus sp., Streptomyces sp., Bcillus sp., Penibcillus sp., nd unidentified fungi.... Winwright et l., Blloon Cryosmpler comprised of sterile cylindricl probes Bcillus sp., Stphylococcus sp., Engyotontium sp..... Griffin,, Aircrft Sterile impcter pltes exposed on the underside of ircrft Bcillus sp., Penicillium sp., Micrococccee nd Microbctericee, Stphylococcus nd Brevibcterium unknown unknown unknown unknown Yng et l., b Blloon Drwing ir through membrne filters by pumps Bcillus sp., Penibcillus sp.... Refer to the references in ref.. bair density t the respective men smpling ltitude ws used for conversion between ATP mbient temperture nd pressurend STP stndrd temperture nd pressure nd kp. capproximte men density vlues. Vol. No.

Fig. 2 Fig. 3 Fig. 4 cm L/minkm L/min V, A km Fig. 3 Devices used for ir borne dust smpling on ircrft.

OW, outer wll of the ircrft; S, support; IT, intke of smpling tube; C, connector for Teflon tube.

IL, inlet connected to the Teflon tube; DP, dividing pipe; BV, bll vlves; F, filter holders; NV, needle vlve; P, pump; OL, outlet to

3 Fig. 2 Fig. 3 Fig. 4 cm L/minkm L/min V, A km Fig. 3 Devices used for ir borne dust smpling on ircrft. AStructure of the smpling tube ttched to the outer wll of the ircrft. OW, outer wll of the ircrft; S, support; IT, intke of smpling tube; C, connector for Teflon tube. B nd CFlow digrm nd the front pnel of n ir dust smpler ADS, respectively. IL, inlet connected to the Teflon tube; DP, dividing pipe; BV, bll vlves; F, filter holders; NV, needle vlve; P, pump; OL, outlet to Teflon tube connected to the exhust pprtus of the ircrft; R, chrt recorder. A needle vlve ws used to control flow rte. Airflow through filter ws controlled individully by ech bll vlve. Fig. 2 The ircrft used for the microbe smpling experiments. Fig. 4 Picture of the devices used for ir borne dust smpling on ircrft.

Strtosphere. Troposphere c.. c... b Airport ABB- d. b University cmpus ircrft d. b ABB- d. b Fig.

4 km L km. μm. μm Fig. 5 km V W.μm mm.m.m Fig. 6 kg kg,l Fig. 5 A blloon used for the microbe smpling experiment. Tble 2 Tble 3 Tble 2 Vible counts of bcteri collected from high ltitude nd ground Site Altitude km Volume of ir Vible count L CFU CFU / m Strtosphere. Troposphere c.. c... b Airport ABB- d. b University cmpus ircrft d. b ABB- d. b Fig. 6 Picture inside of the gondol showing the device used for the microbe smpling experiments. Corresponding mss volume of litter t tm. b Fungous colonies were observed. c Yer of the smpling. d Type of the smpler used. Vol. No.

5 Tble 3 UV resistnce of bcteri isolted from ir dust smples nd of Deinococcus rdiodurns strins Clone No. Altitude km or site D J m D J m ST b b TR.. c c TR. TR.. TR.. AP * Airport AP * Airport AP * Airport TU * University cmpus TU * University cmpus D. rdiodurns JCM D. rdiodurns ATCC d D. rdiodurns ATCC e.... D-vlues re defined s the UV dose which reduced cell popultion to specified percentge of the originl number of cells. The D-vlues were clculted from the regression line of the exponentil slope of the survivl curves s described in Arrge et l. b Rnge of vlues. c MenSEM ormx-men. d Arrge et l. e Gscon et l. * The strins found most resistnt to UV irrdition mong ground-level bcteril isoltes Tble. No. of exp. Fig. 7 UV resistnce of Deinococcus species. Filled circles: Deinococcus rdiodurns. Tringles: D. erius. Squres: D. etherius. Fig. 8 Gmm ry resistnce of Deinococcus species. Filled circles: Deinococcus rdiodurns. Tringles: D. erius. Squres: D. etherius. Fig. 7, Tble Deinococcus Deinococcus Deinococcus Deinococcus erius Deinococcus etherius, Deinococcus Deinococcus rdiodurns rdiodurns D. rdiodurns D. erius D. etherius Fig. 8 Bcillus Penibcillus Tble

km km km/ km/ km/, Fig. 9 Fig. insert μm km/ mm DNA DNA km/ Fig. 10 Tble /m cmcm km/ / Fig. 9 Two stge light gs gun nd the polycrbonte bullet insert. Fig. 10 Microbil density depending on ltitude.

6 km km km/ km/ km/, Fig. 9 Fig. insert μm km/ mm DNA DNA km/ Fig. 10 Tble /m cmcm km/ / Fig. 9 Two stge light gs gun nd the polycrbonte bullet insert. Fig. 10 Microbil density depending on ltitude. Horizontl lines or filled circles were drwn bsed on the dt t mbient temperture nd pressure. Becuse some microbil smplings were done in certin height rnges, the horizontl lines show the respective height rnges. The dshed regression line d/h indictes the estimted microbil density dversus height h. Vol. No.

7 HTV HTV JEM JEM JEM JEM JEM DNA PCR References Yng, Y., Yokobori, S. nd Ymgishi, A.: Assessing Pnspermi Hypothesis by Microorgnisms Collected from the High Altitude Atmosphere, Biol. Sci. Spce, 23, - Yng, Y., Ithshi, S., Yokobori, S. nd Ymgishi, A.: UV-Resistnt Bcteri Isolted from Upper Troposphere nd Lower Strtosphere, Biol. Sci. Spce, 22, - Iijim, I., Izutsu, N., Fuke, H., Sitoh, Y., Kwski, T., Mtsuzk, S., Nmiki, M., Oht, S., Toriumi, M., Ymgmi, T., Ymd, K., Seo, M., Ymgishi, A. nd Yokobori, S.: Development of the Microbe Smpling Device, JAXA-RR-05-, p. -, ISDN- in Jpnese Yng, Y., Itoh, T., Yokobori, S., Ithshi, S., Shimd, H., Stoh, K., Ohb, H., Nrumi, I. nd Ymgishi, A.: Deinococcus erius sp. nov., Isolted from the High Atmosphere, Int. J. Syst. Evol. Microbiol., 59, - Yng, Y., Itoh, T., Yokobori, S., Shimd, H., Ithshi, S., Stoh, K., Ohb, H., Nrumi, I. nd Ymgishi, A.: Deinococcus etherius sp. nov., Iisolted from the Strtosphere, Int. J. Syst. Evol. Microbiol. in press Yng, Y., Yokobori, S. nd Ymgishi, A.: Bcteril Survivl in Response to Desicction nd High Humidity t bove Zero nd Subzero Tempertures, Adv. Spce Res., 43, - Yng, Y., Yokobori, S., Kwguchi, J., Ymgmi, T., Iijim, I., Izutsu, N., Fuke, H., Sitoh, Y., Mtsuzk, S., Nmiki, M., Oht, S., Toriumi, M., Tnd, K., Seo, M. nd Ymgishi, A.: Investigtion of Cultivble Microorgnisms in the Strtosphere Collected by Using Blloon in, JAXA-RR-08-, - Ymgishi, A., Yno, H., Okudir, K., Kobyshi, K., Yokobori, S., Tbt, M. nd Kwi, H.: TANPOPO: Astrobiology Exposure nd Micrometeoroid Cpture Experiments, Biol. Sci. Spce, 21, - Ymgishi, A., Yno, H., Okudir, K., Kobyshi, K., Yokobori, S., Tbt, M., Kwi, H., Ymshit, M., Hshimoto, H., Nrok, H. nd Mit, H.: Tnpopo: Astrobiology Exposure nd Micrometeoroid Cpture Experiments, ISTS 2008-k- Arrge, A. A., Phelps, T. J., Benoit, R. E., Plumbo, A. V. nd White, D. C.: Bcteril Sensitivity to UV Light s Model for Ionizing Rdition Resistnce, J. Microbiol. Methods, 18, - Gscon, J., Oubin, A., Perez-Lezun, A. nd Urmenet, J.: Sensitivity of Selected Bcteril Species to UV Rdition, Curr. Microbiol., 30, -

技術研究報告第２６号

技術研究報告第２６号 1) 2) 3) 250Hz 500Hz RESEARCH ON THE PHYSICAL VOLUME OF THE DYNAMIC VIBRATION RESPONSE AND THE REDUCTION OF THE FLOOR IMPACT SOUND LEVEL IN FLOORS OF RESIDENTIAL HOUSING Hideo WATANABE *1 This study was