嫌気微生物群集による ポリ塩化ビフェニル分解活性の獲得
Contents
1. Monitored Natural Attenuation2. PCB AREY E PCB
3. 4 2 6 B56 in situ HISH Fluorescent in situ hybridaization FISH Amann et al 1990 Daims et al 1999 Yoshida et al 2005 1 ml 30 mM PBS 30 mM NaHPO 30 mM NaH PO 390 mM NaCl pH7 2 sterilized 10 mM CAR LERT 3 2 g dH20 65 mL 6N NaOH 2 ml TRAL KET 2 10 mM PBS fk CUR 1 ml PBS 100
4. ECD PCB 2 Scheffe F F
5. 2005 Rivers amp Reactive Bio barrier Fig 1 2 Conceptual illustration of the PCB decontaminating reactive bio barrier 3 13 Polychlorinated biphenyl PCB 1 3 1 PCB PCB 2 1 10 Hig 1 1 PCB iX FOE 209 100 PCB PCB
6. PCB Ahmed and Focht 1973 PCB 2 10 1 4 1 PCB PCB PCB
7. PCB i Fe mu xa es Ay E PCB tA E E BS Natarajan 1996 3 E Brown 1987 PCB aj IN z A BC iB LC 33 0 meta BAR para 800732 v 53b 5 cx
8. 41 cv Table 3 3 Average residual amount of individual PCB congeners mol in Kuridashi PS and BS cultures after 56 days of incubation during the maintenance of the anaerobic PCB degrading activity over 15 times of the serial transfers IUPAC Ls mol in initial PCB Kuridashi PS Kuridashi BS Significant Substituted positions of Cl i number mixture Residual amount SD Residual amount SD difference 6 2 3 0 6 137 4 22 5 66 6 42 1 8 2 4 4 5 87 8 10 9 42 6 20 8 16 23 2 0 3 98 3 12 6 67 2 329 20 33 45 23 3 34 2 236 2 7 9 83 8 11 8 56 7 17 3 17 24 2 2 8 103 4 14 6 61 0 18 99 28 53 24 4 25 26 10 1 81 1 10 2 51 1 6 18 25 2 7 7 883 11 9 56 7 17 3 FEF 26 50 25 3 246 2 9 6 84 4 11 2 52 9 17 1 31 25 4 9 6 844 4 11 2 52 9 I7 32 26 4 2 0 97 6 11 4 63 9 19 8 44 23 25 6 1 883 14 0 64 1 15 5 46 23 26 3 2 92 8 17 6 61 9 19 2 49 43 24 25 235 2 4 2 84 6 10 3 63 6 18 7 66 24 34 5 8 82 1 10 6 62 4 14 4 52 69 25 25 246 3 6 6 87 2 10 3 729 19 9
9. 92
10. 21 GC ECD f amp H HH ECD FTID ECD CB GC ECD PCB GC ECD PCB KC 300 400 IUPAC 199 Kyoto Japan 4 PCB KC 300 400 100 ug
11. x7 ATSC PCB PCB PCB LPL d
12. TCE PCE 2005 ISO14000s 1 2 2000 2005 2005 3 30
13. 63 Table 4 2 Effects of particle seize on the total residual PCBs in second transferred cultures from GBO 5 culture Conditions L GB2 0 GBO 5 GB0 05 Material Particle size Non GB 2 0 GB 0 5 GB 0 05 Residual amount B 16 20025 690 85 396x4 706 66 3 43 4 97 0 3 8 weight Abbreviations PS paddy soil L liquid GB glass beads The condition names were explained in materials and methods The experiments were performed more than triplicate Significant decrease in the total residual weight was compared with GBO 5 culture by analysis of variance F test followed by a Scheffe multiple comparison test The symbol denote the significant differences at less than 0 1 of probability respectively L GB0 5 GB0 05 GB2 0 LC T T 0 60 120 180 Fig 4 1 Dendrogram from the cluster analysis grouping the DGGE samples of soil free cultures after incubated in GB0 5 culture using Ward method normalized data The scare was corresponded to squared distance Analysis was referred from http aoki2 si gunma u ac jp BlackBox BlackBox cgi 4 3 3 GB0 5 PCB GB0 5 PS PCB
14. Ts PCB 196 4 nmol Cl mmol e donors 8 PCB E PCB 93 9 PCB 8 mmol 56 PCB
15. REPER LOS He St CAE TS S v BE rRNA electrophoresis Desulfitobacterium uu E BER PCB PCB 4 E PCB 16S PCR DGGE Polymerase chain reaction Denaturing gradient gel Firmicutes hh Dehalococcoides BO 3 TRES Dehalobacter E
16. 3 PS BS 3 PCB PCB Boyle et al 1993 Hartcamp commandeur 1996 Cutter et al 1998 2001 Wu et al 1998 2002 Bedard et al 2006 Natarajan 5 1996 1998 PCB 2 PS LOC PS 3 PS BS
17. Fig 1 4 Pseudomonas Alcalegenes Nocardia J Acinetobacter Table 1 8 PCB Wiegel and Wu 2000 PCB PCB Aerobic degradation e acceptor COOH O o lon CO 6 3 y A C H H O Less chlorina
18. PCB I3 KC 300 400 A4 SAK PCB PCB 32 Wiegel and Wu 2000 Hudson River Silver Lakes ezz pg7g PCB Brown et al 1987 ortho PCB Baltimore Habor Berkaw et al 1996 Woods Pond
19. PCB PCB PCB PCB 40 mol Firmicutes PCB PCB PCB
20. f PCB JIS K0093 2 HAF EU EO HE ZE p VINA i PCB PS FREE 1 GC
21. 550 C 24 pH 2 5 39 7 mol PCB 60 3 mol PCB 87 moal 56 E PCB 2
22. E PCB WIES 3
23. Shibata et al 2006 LTO Z EERE SNE PCB KC 300 400 1 1 PCB 42 44 PCB PCB 1
24. 3 2 3 2 1 PCB PCB 2 PCB KC 300 KC 400 Kanegafuhi Chemical Industry Osaka Japan 1 1 PCB 1 gl1 PCB 2 7 50 3 39 9 w w 4 44 4 w w 3 2 2 gley lowland soil FAO UNESCO classification Eutric Fluvisol gray lowland soil Eutric Gleysols ando soil Haplic Andosol 3 2006 PCB PS 2 mm
25. fig 4 9 4 4 PS L GB0 S GB0 5 GB0 5 L GB0 05 GB0 5 GB2 0 PCB
26. yo Se BY He E Flexible vinyl chamber TYPE A Coy Laboratory products Grass Lake MI USA h FECL FOR 80 15 1 4 9 3 5 g Paddy soil PS 4 ml 121 C 30 min 3 100 ug PCB 100 ul PCB VRA g L E AL 15 ml AL PS Paddy soil HF HH LE PCB
27. PCB Voice and Weber 1983 Lyytik inen et al 2003 Lee and Batchelor 2004 BS PCB E JW 52 PCB PS
28. 56 B E PCB 3 50 100 ug PCB PCB gt Qo E F 14 mol E PCB pH E HH PCB
29. 56 PCB PCB Choi et al 2003 FEW O PCB PCB 2 Garbarini and Lion 1986 OPS BIL EIL PCB
30. Biostimulation MERL Team 2002
31. Table 4 6 Effects of electron donors inhibitors and electron acceptor in two different medium on GB0 5 culture AL medium SF medium Electron donors Acetate Formate Lactate Pyrvate Acetate gt Acetate Lactate Inhibitors Heat treatment Molybdate BES Electron acceptor 61 Symbol marks corresponded to the activity compared with the PS culture lt 1 lt 5 gt 10 increased The components of AL and SF medium were described on materials and methods on chapter 2 and 3 BES is abbreviation of 2 bromoethane sulfonic acid Effects of inhibitors and electron acceptors were performed on GBO 5 culture amended with acetate lactate as electron donors 4 8 7 GB0 5 AL PCB 23 1 8 w w 25 746 2 mol 75 ug PCB Fig 4 8 PCB PCB 6 8 17 18
32. 3 2 13 16S rRNA PCR Dehalococcoides 793F 946R 2005 Dehalobacter 441F 645R Desulfitobacterium 406F 619R Smits 2004 Dehalococcoides JE VR EO o 17 DF 1 Chloroflexi 348F 884R Fagervold 2006 PCR 2 39 3 3 3 3 1 2 PS pH ium dcn FE PS
33. 2 situ Fig 1 2
34. 2 1 PCB PCB HT8 0 22 mm ID 50 m L 0 25 um FT SEG Austin TX GC ECD Ni ECD GC 2014 Shimadzu Kyoto Japan Shimadzu Class GC Station Shimadzu Kyoto Japan PCB KC 300 400 100 ug PCB 35 7 ng 42 PCB IUPAC 56 60 PCB 153 180 2
35. PCDF PCDD PCB 80 pg TEQ g fat PCB 2005 1972 PCB PCB 1973 PCB PCB Table 1 3
36. 3 10 50 80 100 3 KETIA Bacteria 16S rRNA EUB338mix 5 gct gcc tec cgt agg agt 3 Archaea 16S rRNA ARCO15 5 gig cic cec cge eaa tte ct 3 NonEUB 5 act cct acg gga gge agc 3 Cye3 3 10 ul 1 uu 30 ng pl 5M NaCl Sterilized 180 ul 1 M Tris HCI Tris 12 1 g pH 7 0 20 ul Formamide 200 pl EUB388 dHO 800 ul EUB388mix 600 ul 10 SDS 2 ul 50 ml
37. Voice and Weber 1983 Lee and Batchelor 2004 PCB 1 PCB PCB H PCB
38. GB0 5 5 56 3 PCB BS PCB PCB 3 PS 5 1 BS 3 4 GB0 KC 300 400
39. PCB 17 2 PCB 2 1 3 RE PCB OP 4 6 CB EE ETE 4 10 30 weight PCB 50 150 ug PCB 4 ED Be BERE Cit E
40. E WHO 2006 sts E 1 TEF 2005 PCB Table 1 6 Table 1 6 The PCB concentration for individual chlorinated congeners in Chlorinated congeners Monochlorinated biphenyls Dichlorinated biphenyls Trichlorinated biphenyls Tetrachlorinated biphenyls Pentachlorinated biphenyls Hexachlorinated biphenyls Heptachlorinated biphenyls Octachlorinated biphenyls Nonachlorinated biphenyls Total PCBs Hit 14 FE POPs 3 the environments in 2002 Concentration in environments Waters ND 0 06 pg ND 9 1 pg 0 02 7 5 pg l 0 02 25 pg l ND 9 6 pg l ND 0 27 pg l ND 0 03 pg l ND 0 02 pg l ND 45 pg l Abbreviations ND not detected 0 01pg l for waters 0 01ug kg for sediments E 2005 8 Sediments ND 0 31 pg kg 0 02 16 ug kg 0 11 110 us kg 0 26 130 ug kg 0 37 93 ug kg 0 26 55 ug kg 0 06 30 ug kg ND 14 pg kg ND 0 63 ug kg 1 2 430 ug kg E
41. SF SF BS 196 LEZ Uw 2 TZA PCB PCB PCB PCB BS 28 KC 300 400 25 ug g BS 6 7
42. Table 1 2 Correspondences of the commercial PCB mixture Kanechlor Aroclor Dichlorinated biphenyls KC 200 Aroclor 1232 Trichlorinated biphenyls KC 300 Aroclor 1242 Tetrachlorinated biphenyls KC 400 Aroclor 1248 Pentachlorinated biphenyls KC 500 Aroclor 1254 Hexachlorinated biphenyls KC 600 Aroclor 1260 Table 1 3 Environmental standards for PCBs in Japan Environments Environmental standard Foods fishes lt 3 ppm Soils ND Waste waters lt 30 ug l Drinking water and ground water ND Sediments lt 10 ppm Atmosphere lt 0 5 ug m Exausted gas X0 15 mg m Abbreviations ND not detected 0 01pg L for waters 0 01ug kg for sediments 14 FE POPs 2005 1 3 3 PCB PCB 1 15 PCB PCB PCB PCB E HH
43. Development and characterization of anaerobic consortia degrading polychlorinated biphenyls LOW fe M 1 UIN salt ol a PP ET 1 1 2 oo 2 EI A VREE ao IA ern Cnr AR RORRUR T ee eet Te er RT reer DR 4 1 4 PCB ERE 10 1 5 me 15 2 PCB 18 Ti a aioe aed aah m T M EE 18 22 FER TT E 19 UP T 26 D ea ee ne ee eee E ramet Oe error eee etree ere 31 3 PCB ttt 34 CIE Uc MAL NOURRITURE M T E UR 34 LN yi m 35 QR cr p 40 3 52 4 3 PCB 58 58 4 2 59 TEE e E ER RR RR ER 62 BAS EI hic Bala a EE Rl ht Pat SF GE 75 78 1 c 78 5 0 DED
44. 2 PS pH PCB Wiegel and Wu 2000 Wu et al 1997 PS 3 E ERO pH Wiegel and Wu 2000 6 PS PCB
45. 37 PCB 15 425 2 42 PCB 2 44 PCB Kim KC 300 400 83 0 ww PCB 100 ug 92 9 5 5 w w 93 6 PCB PCB PCB PCB Bonferroni F tesD Aspin Welch
46. 41 0f ef S AR 1 60 ug PCB SY ETE 3 E 77 5 5 1 PCB PCB PCB IRE DURAND ARTES SLM LT PCP Yoshida et al submitted Fthalide Yoshida et al unpublished
47. BS PS 550 C 24 BS 56 PCB PS 2 5 15 3 PS PCB BS KC 300 400 PCB BS PCB BS PS DPS GPS BS PCB
48. GB 2006 PCB PCB ul 80 4 E PC
49. PCB 100 ug 7 8 mmol ERG ERES SWB LTRS DEV RBS H hie Cla 0 017 Bouchard et al 1996 Mohn and Kennedy 1992 Utkin et al 1995 54 BS Firmicutes P8 Deltaproteobacteria RC Epsilonproteobateria 16S rRNA PCR DGGE Firmicutes Clostridia Fig 3 5 BS Finmicutes PS PCB Firmicutes FY Bedard et al 2006 Yan
50. 4 PCB ij PCB E PCB PCB PCB PCB 31 PCB Tiedje et al 1993 Borja et al 2005 Wiegel and Wu 2000 PCB HA Yoshida et al submitted Fthalide
51. 3 WEI e 10 cm 30 cm 2 mm DOMENI LER 22 C 2 PCB PCB 2 2 3 PCB 180 C 3 hour 600 ml 17 g 20 ml 300 pl PCB 1 gL 80 ml
52. 30 C 4 6 mineral salts MS acetate lactate AL 2 MS g D K HPO4 0 27 KH PO 0 35 NHj CI 2 7 MgCL 6HO 0 1 FeCl 4H O 0 02 CaCb 2HzO 0 1 Yeast extract 2 0 1mg7I pH tk KOH 7 0 L 100 mM MS AL g K HPO 0 5 NH4 SO 0 5 MgSO 7H 0 0 1 FeSO 7H 0 0 02 Yeast extract 0 5 8 10 AL Il mg L pH KOH 7 0
53. 18 9 ng 10 0 FT PCB Kim 2004 KC 300 400 83 76 6 PCB PCB 100 92 9 5 5 PCB PCB 100 PCB weight PCB PCB PCB PCB mol mol
54. SUS Porarpak Q ZP 20 80 100 3 0 mm inner diameter 3 0 m length Waters Milford MA USA GC FID GC 14B Shimadzu Kyoto Japan 1 0 mlmin 30 mllmin 140 C EL 70 C H 100 ul 0 1 mol 2 2 9 DNA PCR DGGE DNA ISOIL extraction kit Nippon gene Tokyo Japan ggcrerig 16S rRNA 341F 5 cgc cgg Ccg cgc gcg gcg ggc ggg sce ggg gca Cgg gggg cct acg gga ggc age ag 3 GC clamp 518R 5 att acc geg get get gg 3 rRNA 341 518 Myzer et al 1993 PCR 200 pl 100 Takara Bio Shiga Japan
55. PCB 3 PCB K F s RETZ E o TRF Aspin Welch 3 B 22 AR o Scheffe 2 2H EE WREE Eo TH E 1 GC ECD ECD N
56. VEO PCB PCB PCB PCB 2005 PCB E PCB
57. T dt rf 6 PCP Pentachlorophenol Yoshida et al unpublished Shibata et al 2000 0 Yang et al unpublished PCB PCB ft PCB 10 1000 pg 100ml culture PCB MEK PS
58. 3 E 15 2 5 56 GB0 5 PCB 58 4412 0 mol P lt 0 01 39 0 10 7 mol P lt 0 01 3 BS PCB 56 GB0 5 PCB 3 PS Table 4 3 PS 852 169 6 18 103 P lt 0 01 PS 64 Table 4 3 Average residual amount of individual PCB congeners mol in GB cultures after 56 days of incubation during the maintenance of the anaerobic PCB degr
59. Student Aspin Welch Chocran Cox Aspin Welch 5 2 2 7 100 ul ZORBAX ODS 4 6 mm inner diameter 250 mm length Wilmington DE USA 2 thigh performance liquid chromatograph equipped with a photodiode array detector 9 2 1 0 m min 275 nm 270 nm Katayama and Fujie 2000 23 2 2 8
60. E ot PCB E E 5 E 2 E 1 6 2001 4 PCB TEs 4 6 En g LIU A Lua
61. PCB Table 1 4 JESCO Japan environmental safety corporation 22 95 5x00 4E7U M 2006 20 Ah PCB E 5 PCB Table 1 5 ESCO Table 1 4 The methods for decomposition of PCBs
62. 6 13 5 mol 5 2 mol 14 1 mol 15 1 mol 5 2 mol 4 mol AL 6 PCB me mil Zu oY 26 Table 2 2 Total residual PCBs in Kuridashi paddy soil cultures s
63. PCB i PS GB0 5 PCB 4 0 5 mm GB BS 3 GB PCB PCB GB fr
64. BS PCB Kim and Rhee 1997 Wu et al 2002 Zwiernik et al 1997 56 BS PCB E PCB PCB Wiegel and Wu 2000 Ye 1999 BES BS BES PCB
65. situ exsitu 2 In situ ex situ 2000 2 Bioaugumentation
66. Firmicutes Bacteria PCR DGGE Lot 34 E Archaea DGGE PCB PCR PCB PCB
67. Scheffe n non parametric Bonferroni Scheffe 3 Aspin Welch t t 2 2 t F
68. 2 2 10 DNA PCR Wizard Plus Minipreps DNA purification System Promega Madison WI USA DNA pTBlue Perfectly Blunt Cloning Kit Novagen Madison WI USA DNA Templiphi plasmid amplification kit Amersham Bioscience Sunnyvale CA USA DNA 2 UV 16S rRNA DNA ABI 3100 Applied Biosystems Foster CA USA DNA GENETYX Version 7 Software Development Co Tokyo Japan NCBI BLAST 24 BLAST http www ncbi nlm nih gov BLAST 16S rRNA ERR ZITOE Table 2
69. 1 E fes IR BE DHE D C Kv VERRY PCB Fig 1 2 1 4 PCB PCB
70. 27 E PCB E Table 2 3 Effects of inoculation rate and incubation interval on the total residual PCBs in Kuridashi paddy soil cultures Inoculation rate Incubation Total residual PCBs 96 v v interval days 96 w w 0 56 101 3 1 56 100 0 5 56 86 8 10 56 79 4 50 56 70 T 5 0 100 0 5 14 99 9 5 28 92 4 5 56 86 8 Total residual PCBs are shown as w w of the amount in the sterilized control after the same incubation time The values are the average of between two and seven determinations using separate incubation vials Statistical significance of amounts of total residual PCBs compared with sterile control was examined by analysis of variance F test followed by a Scheffe multiple comparison The symbols and denote a significant difference at lt 5 lt 1 and lt 0 1 respectively PCB PCB 36
71. Table 3 5 PCB 48 ll Fig 3 5 The DGGE fragment patterns of the amplified 16S rRNA genes for Bacteria a and Archaea b from microcosms of the four cultures after 56 days of incubation Lane 1 shows the profile of the original Kuridashi paddy soil Lane 2 shows that of the paddy soil PS culture Lane 3 shows that of the burnt soil BS culture Lane 4 shows that of the BS culture amended with 2 mM of molybdate Lane 5 shows that of the BS culture amended with 2 mM of BES The bands numbered in circles were subjected to the DNA sequencing and the phylogenetic analysis as shown in Table 3 5 Table 3 5 Phylogenetic assignment of the DGGE clones of partial 16S rRNA genes isolated from the microcosm in the BS culture Band Phylum class Closest relative 1 Firmicutes Uncultured bacterium clone Z39 2 Firmicutes Uncultured bacterium clone IIIA 3 3 Firmicutes Clostridium butyricum Strain EIB3 3 4 Firmicutes Uncultured bacterium clone BF11 5 Deltaproteobacteria Desulfovibrio vulgaris DAM644 6 Chloroflexi U
72. BS PS BS f BS upflow anaerobic blanket reactor UASB 4 E Sekiguchi et al 1999 Zheng et al 2006 BS PCB 4 BS BS AL PCB 4 7
73. 196 H EEH 0 14 mmol FLER 0 03 mmol 0 002 mmol H amp E 0 001 mmol 21 mol 0 2 mol H PCB 45 1 2 8 w w 43 5 2 8 mol 54 9 mol SF BS 45 GSE CEA PCB Care H EB 100 O oo 70 Total residual PCBs w w 0 20 40 60 80 100 e without addition with addition 120 140 160 180 200 Incubation period days Fig 3 3 Effects of sulfate free medium on the total amount of PCB residues w w in Kuridashi BS culture over 196 days of the incubation with or without additional acetate lactate electron donors with appropriate intervals Arrows show the time that the
74. 3 AL PCB Firmicutes Chloroflexi PCB PCB B BS PCB AL 74 3 mol 75 ug E Firmicutes TT AL Nath and Das 2004 Fe
75. PCB E La 2 f ZJE ayy tA E E E E PCB E gt eG gt eG N iX 56 13 rRNA 7 OAD T2 H H gt Ge Epsilonproteobacteria Firmicutes FH Deltaproteobacteria Chloroflexi 4
76. GB0 5 PCB AL 72 A E Sterile control IIIA D 1 A A IS 7 Er 5 LZ Q c le oo ANA 9ET OVET VE VE SVC 9ET 9ET 9ET SVC VET ve 9c Ssc 9vc ec S Cc 99 SSC CSVECIV OSETIET SVT E 9c VE VET V SVT CVEC V 9ET SE ST VE Se 9VC SC ST Sc 6 9 C SC voc 9 V SC C9OVC E ST TST 9c Scr vc eV TYT c otcocvtce tc TET rv rc CG e dures 8n 3unoure enprsos 73 Compositions of congenrs Fig 4 8 The residual amounts of the PCB congeners in the GB0 5 culture amended with 15 ml of AL medium after 3 months of incubation and incubated for 56 days The residual amounts of individual congeners were compared with the sterilized culture Numbers indicated under the figure correspond to the structure of individual PCB congeners
77. FFirmicutes Yan et al 2004 Bedard e al HAF 8338 LE EIA zzcz7esy E Sokol et al 1994 Rysavy et al 2005 SF SF AL E PCB fc E GB0 5 76 MEIC E 3
78. PCB GB0 5 PS BS Chloroflexi 4 h 4 E ER a Ze Eb E Halt MPR YU HRT KU E 16S rRNA DGGE PS BS Firmicutes 4 E 4 Firmicutes E 4 E 2006 GB PCB GB0 5 PCB E E PCB PCB
79. Firmicutes Desulfitobacterium Desulfitobacterium KERIK PCB 3 3 5 5 tetrachloro 4 4 biphenyldiol iii Wiegel et al 1999 Krasotkina et al 2001 Chloroflexi Dehalococcoides PCB PS Fennell et al 2004 PCB 33 1D Firmicutes 3 PCB 3 1 2 PCB 56 PCB 14 mol
80. NIWA 1 NIWA 3 3 cm TYK 1 4g PCB L 5 V y 300C NIWA 1 NIWA 3 PCB AL PCB TYK 1 A3 PCB 36 PCB PCB 3 2 8 PCB PCB
81. BS BS GB PCB PCB PCB GB Fava 1996 65 45 573 PCB PCB PS PCB
82. Bes CA S OO PCB 3 PCB PCB LAM E SE GB0 5 amis EO S GB0 RIC MRA a GB0 5 PCB gt EZ BES FAY PCB BB LTe Pork lk Pb PCB E Firmicutes P Cutter et al mu 200
83. PCB 93 9 nmol Cl mmol e donors 3 3 4 BS PCB 196 Fig 3 2 BS 36 PCB 196 3 E BS E BS PCB S6 PCB PCB 10 w w PCB 56 43 Total residual PCB w w Acetate amp Lactate O Sulfate mmol culture mmol culture Amount of sulfate Amount of acetate lactate cells ml Microbial population
84. PS BRAW E LT 5 vy 36 PS 5 w v BAT 5 ELT 2 PS PS PCB WTKR de BE 0 1 PCB PS 0 1 S 10 50 28 PCB
85. EOD FR PCB 3 2 GEN 1H PCB 2 5 PCB 3 PCB
86. PCB PCB 1000 ug 36 60 ug PCB gt ay _ t 42 4 2 1 PCB 2 3 39 9 w w 4 44 4 w w 50 PCB PCB 58 Kanechlor 300 400 Kanegafuhi Chemical Industry Osaka Japan 1 1 1 g l DTE hE 4 2 2 Qlass beads G
87. PCB 1 Bedard Bedard et al 1996 1997 2006 Bedard and Quen sen 199 PCB Td meta pg7g ortho p TE TH 79 B Ae 1 ES Natarjan 1996 Bedard 2006 f PCB S 3 BB MEDS HER C amp C
88. 6 16 17 32 100 erz pg7g ozzo Z PCB PCB a 28 Table 2 4 Residual amount of individual PCB congeners mol in Kuridashi paddy soil cultures after 56 days of incubation averaged over 13 serial transfers of an anaerobic PCB degrading culture mol in Residual s Substituted positions of Cl initial PCB amount SD NH mixture mol 6 2 3 1 0 114 8 168 8 2 4 53 849 268 16 23 2 27 669 352 20 33 45 23 3 34 2 236 2 7 2 81 3 92 we 17 24 2 2 8 101 6 133 28 53 24 4 25 26 9 9 823 t 97 we 18 25 2 7 8 93 7 109 26 50 25 3 246 2 1 1 91 8 21 8 31 25 4 9 9 853 86 32 26 4 2 2 98 4 162 44 23 25 5 6 87 1 98 s 46 23 26 7 6 8
89. PCB PS PCB 3 BS 8 w w 3 FEO BS f PCB GBS PCB 76 3 5 2 w w LIVEN 88 4 10 2 w w 91 2 13 5 w w PS 40 BS PS DPS GPS PCB Bx L Table 3 2 Chemical properties of PS and three BSs and the total PCB residues in the cultures containing PS and BSs as carriers after 56 days of incubation PS BSs Properties Kuridashi Kuridashi Kamashima Nagakute Total carbon 1 0 ND ND ND Exchangeable Mg mg kg dry soil 0 63 29 6 76 1 3 3 Exchangeable Ca mg kg dry soil 178 7 202 5 392 5 31 7 Exchangeable K mg kg dry soil 2 4 110 0 145 0 25 1 Exchangeable Na mg kg dry so
90. 6 7 43 48 F Deltaproteobacteria W Epsironproteobacteria Firmicutes Katayama and Fujie 2000 Bacteria 2c 16S rRNA PCR DGGE L7 Fig 2 1 PS DGGE 1 8 9 13 DGGE 168 rRNA Table 2 5 9 DGGE 6 Ald Firmicutes Clostridia pgc7e7o7Ze7es e zz 7o7eo gc7e77g Chloroflexi Firmicutes 6 7 Firmicutes PCR 3 Dehalococcoides Dehalobacter Desulfito
91. 9 1 9 1 3 2 2 3 8 2 8 2 4 4 7 a f 3 6 O 4 e 9 4 5 op 6 5 PCDDs PCDFs Polychlorinated dibenzo p dioxin Polychlorinated dibenzofuran PCBs Polychlorinated biphenyl Fig 1 1 Chemical structure of polychlorinated dioxins PCDDs PCDFs and PCBs 1 2 Bioremediation
92. GB0 5 PS GB0 5 1 2 2 UC HD PO PCB PS PCB Scheffe BE nu 59 F tesD Aspin Welch GB0 5 AL SF 3 20 mM
93. PCB PCB PCB ADI Acceptable daily intake 0 005 mg Kg day 10 20 sg 50 kg 0 5 g iR 50 kg C 2 g 50 kg 2005 LogKow 4 3 8 26 BCF Bio concentration factory 120 000 270 000 2004 4 4 PCB Table 1 1 T
94. B lt DR 60 100 PCB PCB Scheffe F tes Aspin Welch PCB nmol CD mmol e donors 1 PCB http aoki2 si gunma u ac jp BlackBox BlackBox cgi 1
95. BS BS 1 PCB Fig 3 7 BS PCB EI efg pg7g oz7o BS 55 KC 300 400 RE OOOO qc cl CC 66 pa meta re f para meta Vara meta Cl E Cl 16 E 25 828 ll ortho J J ortho Cl CI see S 15 U U Fig 3 7 Possible dechlorination pathways in Kuridashi BS culture Numbers indicated under individu
96. 20 mM 5 H 20 mM 20 mM 2 bromoethane sulfonic acid BBS 2 mM 100 C 10 PCB 50 uM 6102 3 4 PeCB 20 mM PCB PS 1 596 10 1 PCB GB0 5 3 AL 15m 56 GB0 5 3 7 AL 15 ml
97. E GB0 5 BacterialArchaea FISH Archaea GB0 05 Archaea 80 0 Bacteria 17 6 SREI GB ERBKE l ROIECRH A OPERIG AK A T LPAR SIV Ele PCR DGGE GB RE 0 5 mm COR PCB GBO 5 PCR DGGE fiir l c X 0 Firmicutes PH Deltaproteobacteria 4
98. BANE CH PCB 3 PCB PCB PCB 6 PCB Table 2 2 MS PCB PCB PCB AL AL
99. Shikkonen and Passivira 2000 4 GB0 5 PCB 100 ug 75 GB0 5 PCB 10 1000 ug 630 AL GB0 5 5 140 100 ug PCB 4 1 75 ug PCB hace Gea or 5 2 PCB PCB BS GB PCB
100. PCB PCB 3 PS 3 4 3 GB0 3 BS PS PCB PCB 3 PCB PCB GB PCB PCB
101. PS 4 ml PS 3 5 g dry weight L lt I4 4 g DPS GPS BS GBS Coy Laboratory Products Grass Lake MD1 g l PCB 100 AL 15 ml 100 PS AL PS DPS DPS GPS Hei BS HiM GBS PCB 5 v y 300C 56 PCB PCB PS PCB Sulfate free SE SF NaCl 1 g KCI 0 5 g NH4CI 0 5 g CaCI 2H50 0 1 g MgCl 6H50 0 1 g yeast extract 0 1 g trace elements SL 10 1 ml S
102. 1 H Table 1 1 C O Rick CLE 60 PCB H Cl E H PES TEQ Toxic Equivalent E 1999 7 PCB PCB 5 6 PCB 3 4 S UN
103. PCB PCB PCB PCB PCB PCB PCB PCB 2 PCB
104. PCB 1999 UV 1100 PCB PCB a Lu HETA fig Table 1 5 Five plants of JESCO and their disposal capacity Places Initiation date Disposal capacity Kitakyusyu Dec 2004 0 5 t day Toyota Sep 2005 1 6 t day Tokyo Nov 2005 2 t day Osaka Aug 2006 2 t day Hokkaido Oct 2007 1 8 t day 1 3 4 PCB 2 3 7 8 TCDD CTEE Toxic Equivalency Factory 2 3 7 8 TCDD 4
105. SUS Packed column J Active carbon 60 80 3 mm ID 2 m L GL Science Tokyo Japan Packed column J Molecular sieve 5A 60 80 3 mm ID 3 m L GL Science Tokyo Japan 150 C 80 C 50 C 100 ul 0 1 mol 3 2 10 PCB 2 1 HPLC 2 3 2 11 PBSE 130 mM NaCl 10 mM 1 mM EDTA pH7 0 2 38 0 2 um 1 ml
106. 7 56 7 PCB E PCB 56 196 PCB 3S6 16 6 mol 0 2 mol 0 1 mol 0 018 mmol 0 010 mmol PCB
107. 1954 E Kanechlor KC KC 300 KC 400 Fig 1 3 1969 Aroclor Aroclorl242 Aroclor1254 Table 1 2 Aroclor ioe PCB 7 Hr 4 Table1 1 Toxic equivalency factors for dioxins for human risk assessment congeners congener TEF PCDDs 2 3 7 8 TCDD 1 1 2 3 7 8 PeCDD 1 1 2 3 4 7 8 HxCDD 0 1 1 2 3 6 7 8 HxCDD 0 1 1 2 3 7 8 9 HxCDD 0 1 1 2 3 4 6 7 8 HpCDD 0 01
108. PCB BS 56 Iu LC 25 Natarajan 5 1996 5 UASB 70 75 Aroclor 1254 50 ug ml 220 ng total PCBs ml culture day Natarjan 10 238 1 93 9 nmol Cl mmol e donors 731 nmol 1 1 PCB 100 ug 0 24 umol
109. 46 C 90 61 50 ml 48 C 1 M Tris HCI pH7 0 Iml 5 M NaCl 9 ml EUB388 2 15 m dH2O 50 ml 48 C 20 20 DAPI 4 6 diamino 2 phenylindole 4 2 7 PCB 2 1 HPLC 2 4 2 8 DNA PCR DGGE
110. Bs E e o Oo 2g i Dt 8 A usd a zm I BL E ERE Ie ERR ri Hr qM E HT N jo oo Nel b N j opdures 3n Junoue TenpIs9 9ET OVET VEVESVC 9ET 9ET 9ET SVC VET ve 9c ec 9vc VESVC 9VC IET ST SVT CSVETIV OSETIET SVT VE 9ET VEVET V SVC CVET V 9ECTI SE ST VEST 9vc Sc Sc VEVC CSEC ST VT Oc tc Sc tc 9c 9 c COVCE ST TST Oc Secr vc an LG CVC C 7 TET vv rc t6 Compositions of congenrs 74 ixture and 15 ml of AL medium after 3 5 culture amended with 1000 ug of PCB m Fig 4 9 The residual amounts of the PCB congeners in the GBO d incubated for 56 days bation an months of incu The residual amounts of individual congeners were compared with the sterilized culture Numbers indicated under the figure correspond to the structure of individual PCB congener PCB 94 0 2 9 w w 95 542 9 mol GB0 5 AL 10 PCB 36 GB0 5 60 ug PCB
111. JL Shimadzu Class GC Station Shimadzu 2 35 7 ng 56 60 PCB 42 fa Eg GC MS KC 300 400 a EO AUG 100 ug ex 20733 45 PCB 7 PCB 28 52 101 4118 4153 4180 Takasuga et al 2005 153 180 2 KC 300 400 ng
112. 61 3 15 9 w w 60 3 16 1 mol P lt 0 01 36 5 13 4 mol P lt 0 01 BS 56 PCB Table 3 3 KC 300 400 PCB PCB PCB 41 72 64 136 176 ez pgzg o777o 74 4149877 PS 6 17 32 BS 46 88 16 BS PS
113. GB PCB PCB GB CIRE 75 L Ht E GBO 5 PCB GB0 5 GB 0 5 mm EO NET Wi ES mm cu E
114. BES BS E f BES PCB ICE OBA Fe O C R mj PCR DGGE Firmicutes itt TYK 1 4 1 35 7 mg 3 4 E TCP Trichloropheno 2006 TCP 1 PCB
115. 7 PCB PCB PCB PCB 7 KC 300 400 PCB 83 PCB IER 5 3 Old BS
116. 1 WIE 2 25 PCB 56 e AES 14 PCB PCB 20 PCB 68000 rpm 10 min 80 ml PCB 2 1 20 ml
117. PCB 35 7 ng IUPAC 56 60 2 PCB 153 180 PCB Kim KC 300 400 83 0 w w PCB 100 ng 94 2 3 5 w w 91 3 PCB PCB PCB 36 HEGRE 10 4 0 5 w w
118. Sokol et al 1994 PCB PCB Ye et al 199 Zwiernik et aL 1998 PCB Wiegel and Wu 2000 pH PCB Bedard et al 1998 13 I Microbial source Bedford Harbor Hudson River Woods pond Housatonic River Silv
119. i NX E PS PS E BS PCB
120. DNA ISOIL extraction kit Nippon Gene Tokyo Japan PCR DGGE PCR ggc7erzg 341F 518R Archaea 344F 518R O 16S rRNA PCR X U DGGE PCR DNA PS 4 DGGE ggc7erig DGGE Fig 4 3 4 2 9
121. TYK 1 PCB Zs ORM bak 23 1 PCB E NIWA NIWA 3 PER OFFS PCB NIWA M PCB PCB 3 EN Firmicute
122. Choroflexi Bedard et al 2006 Fegervold et al 2006 Yan et al 2006 BS Chloroflexi Dehalococcoides Cutter et al 2001 Wu et al 2002 Fennell et aL 2004 BS DGGE 1 FEO Chloroflexi BS PCB BS PCB
123. Bedard Bedard et al 20000 hang et al 1997 Meckenstock et al 2000 Natarajan et al 1999 PCB 1 5 2 15 mill Es 7 E Ho Op K Zu
124. Verg Bum HEAL UT T 56 PCB 56 Ku YLERMR OO BE Chloroflexi ET DEBRIS LOR RIO RIC oC Hi 1 PCB Cutter et al 2001 Cutter et al 1998 Holoman et al 1998 Wu et al 2000 Maym6 Gatell et al 1997 Adrian et al 1998 Z 4 pow 7 2 BS PCB
125. PCR DGGE 56 BS BS 15 3 BS 0 01 umol 6 3 56 pmol 7 0 02 nmol in the culture PS 6 7 0 99 umol 2 02 umo 6 7 Firmicutes F3 Deltaproteobacteria Epsilonproteobacteria Katayama and Fujie 2000 Bacteria KO Archaea 9e 16S rRNA DGGE Fig 3 5 BS Bacteria DGGE PS DGGE Firmicutes Deltaproteobacteria
126. AL 835 E 4 3 E PCB PCB AL Fes 53 SF PCB AL BS d AL
127. BBES DGGE 4 5 6 PCB E 16S rRNA 9 o 17 DF 1 Chloroflexi BUTI EA Joti Bj iF BF 3 3 9 3 NIWA 1 NIWA 3 NIWA 3 Eu HAE Ju Fri 7 PCB Table 3 6 NIWA 1 Cl NIWA 3 E PCB n Nr 15 Kk 11 H KIM 1 Dehalobacter H amp PCB Dehalococcides 7 N H E Desulfitobacterium NIWA 1 N
128. Dehalococcoides Desulfitobacterium 3 4 3 PCB PCR DGGE 1f f TERM E FE ri JATARI HRDLE EI 14 mol PCB PCB E 1A tA EJ E Ei 4 E 3 TUE X7 ves HH Firmicutes Deltaproteobacteria 4 E PCR DGGE 16S ff 6 E
129. PCB nmol C mmol e donors 3 2 9 1 ml 0 45 um 100 20 ul L column ODS 4 6 mm ID 250 mm L CERI Saitama Japan HPLC CTO 10A Shimadzu Kyoto Japan 2 GC FID thermal conductivity detector TCD GC GC 14B shimadzu Kyoto Japan
130. 78 Cutter et al 2001 Wu et al 2002 Natarjan et al 1998 Yan et al 2005 Yoshida et al submitted gt PCB PS PS ba 3 E 2 PS 42 F PCB BS E PS
131. PCB 3 PCB 2001 4 PCB PCB 2016 7 PCB V H 1 E E PCB PCB acl ZW Ji E Wo PCB PCB
132. PCB 56 Fig 3 1 a BS 1 46 mmol 1 34 mmol f cf SLB ERE 7 7 56 0 064 mmol 7 4 E 7 56 Fig 3 1 c PCB
133. 4 Hig 4 4 Fig 4 5 Vik Archaea DGGE pgc e77g DGGE Bacteria DGGE PS GB0 05 Fig 4 6 GB 0 5 PCB 66 ay acera EUB388 mix B Fig 4 2 Fluorescent in situ hybridization of Bacteria using Cye 3 labeled EUB338 DAPI image A and Cye 3 image B of of EUB 338mix probe hybridized cell 67 Fig 4 3 Fluorescent in situ hybridization of Archea using Cye 3 labeled ARC915 probe DAPI image A and Cye 3 image B of of ARC915 probe hybridized cell 68 Fig 4 4 The DGGE fragment patterns
134. BS PCB Itt ESSERE FIO TEC REGE C LUC DOOR E E PCB 81 PCB W Hr A AE XE Ub 1 WT E 2004 PCB 1 384 H
135. PCB PCB Soil free culture 0 5 mm
136. 196 49 1 mol PCB fiesta PCB E SF EEDD MEEN K OO 56 5 mol PCB 4 Ei K 4 E PCB E E
137. 2005 DEA AYE E 341 363 g TEQ 0 059 pg TEQ m 0 33 pg TEQ L 7 5 pg TEQ g 0 063 pg TEQ L ER CI 3 1pg TBQO g PCB Table 1 7 PCB 1 S 2005 3 4 PCB 10 Sinkkonen and Paasivirta 2000 2005 PCB 10 H 38 5 6 PCB
138. 7000t PCB H IPCB E o 10 ppm Table 1 3 PCB PCB PCB PCB PCB 30 PCB 9 y V Hz i PCB PCB
139. Hig 5 1 2 TA HSI B al pen pyruvate Propionate Firmentation by Firmicutes PCB PS 4 HE BS GB0 PCB Fig 5 1 Conceptual illustration of hydrogen utilization in the anaerobic PCB degrading culture PCB PCB 3 ates 300 ug 400 PS PCB 100 ug 240 H 3 BS 82 Tes
140. e7z pg7g PCB ozzo PCB Wu et al 1997 ez pg7g PCB oro PCB PCB PCB PCB PCB 3 PS 4 E E PCR DGGE 16S rRNA Firmicutes Clostridia PCR
141. 94 C 3 min 94 C 30 sec 52 C 30 sec 72 C 1 min 35 72 C 10 min 341F S18R 50 pmol ul each 2 5 U Mg free Extaq DNA polymerase Takara Bio Shiga Japan 10 ul Extaq buffer 20 mM Mg Takara Bio Shiga Japan 10 ul dNTP mixture 2 5 mM each dNTP 1 2 ul DNA template 24 35 ng 74 ul JK WUK PCR DNA DNA 16S rRNA PCR 0 2 ug 40 70 Table 2 1 0 5xTAE 20 mM Tris 10 mM acetate 0 5 mM Na EDTA pH8 0 100 V 60 C 14 SYBER green I Molecular probes Engene OR USA UV DNA
142. PCB 2 bromoethane sulfonic acid BES 2 mM BS 56 BS 3 2 7 3 NIWA 1 LIES R SOPRA RERAN y p T NIWA 3 ot No F050823 TYK 1 3 TYK AL IS ml PCB 100 uug total PCB mixutre 100
143. E PCB PCB T PCB PCB PCB PCB 3g PCB 16 i EL ZOH 3 PCB PCB 4 H PCB
144. Table 3 1 PCB Table 3 1 Effects of incubation conditions on the total residual PCBsin Kuridashi paddy soil cultures Residual Conditions amount SD mol Preparation condition Aerobic 90 9 0 0 Microaerobic 91 0 0 0 Heat treatment 100 C 10min 90 4 8 0 Inhibitors 2 mM BES 88 5 7 6 2 mM Molybdate 88 7 7 5 pH pH 5 89 9 7 1 pH 9 90 6 7 6 Temperature 4 C 93 5 44 22 C 90 3 2 3 Shaking 100 rpm min 90 6 3 0 Reducers 2 5 Cystein HCl 90 9 12 3 2 5 Thioglycorate 91 3 10 7 2 5 Sodium sulfate 90 6 10 0 Serial transfer was performed with and without nitrogen gas flow microaerobic aerobic in the c 3 3 2 BS PCB lean work station Table 3 2 PS 3 BS 56 PCB PCB 98 9 0 5 w w PS BS PCB PS 2 PS
145. F PCB 41 6 mol 40 ug 3 E BS GB0 5 PS PCB 39 13 9 mol GS 14 ng FIJE PCB 39 7 mol 40 ug 44 39 GB0 5 E PCB 4 25 PS GB0 5 2 3 BS PCB 2 fi E BEE gt ee _ CAUSA E 72 ERR ERI 1 PEE ACCU
146. 1000 ug PCB 56 AES PCB uil 4 2 5 PCB PCB 2 PCB 2 1 PCB hexane PCB HT8 0 22 mm ID 50 m L 0 25 um FT SEG Austin TX USA GC ECD Ni ECD GC 2014 Shimadzu Kyoto Japan Shimadzu Class GC Station Shimadzu Kyoto Japan KC 300 400 PCB 100 ug 42 FEO PCB
147. 25 P lt 0 05 BS Fig 3 4 BS Fig 3 4 PCB PCB 46 LV Table 3 4 Average residual mol percentage of each PCB congener in the BS culture amended with 2 mM of molybdate and BES after 56 days of the incubation IUPAC xr MEN BS Molybdate BES a fs mol in initial ET congener Substituted position of Cl r inber PCB mixture Residual amount Residual amount D Residual amount D 6 2 3 0 6 65 8 10 2 137 0 33 9 93 0 13 6 8 2 4 5 1 49 3 10 7 59 8 5 6 67 9 4 7 15 4 4 0 04 44 2 7 3 447 8 7 6 de 60 7 3 3 16 23 2 2 9 67 8 4 4 210 94168 9 84 9 3 2 20 33 45 23 3 34 2 236 2 7 2 67 5x5 6 68 7 9 2 84 2 3 9 17 24 2 2 6 71 0 6 2 85 545 8 86 843 8 25 24 3 0 02 63 5x8 2 679 9 146 9 82 6 4 9 28 53 24 4 25 26 8 9 65 5x8 6 14 6 7 1 Ar 84 4 4 5 18 25 2 8 3 69 7 4 9 67 6 15 7 84 2 3 4 26 50 25 3 246 2 1 1 69 448 3 286 2 219 2 85 8 3 9 31 25 4 8 7 66 1 7 3 95 3x14 9 84 1 4 6 32 26 4 3 5 53 7 6 8 76 3 21 2 31 7412 7 44 23 25 6 4 67 0 2 1 30 5x1 3
148. 3 4 16S rRNA 4 3 4 3 1 PCB Table 4 1 PS 2 PCB PS 2 PS GB0 5 GB2 0 PCB 62 L GB0 05 PCB PCB PS GB0 05 0 P lt 0 01 42 P lt 0 05 66 P lt 0 05 3 GB2 0 103 P lt 0 050 1
149. GB PCB A PCB PCB 4 E 84 Adrian L Manz W Szewzyk U G risch H 1998 Physiological characterization of a bacterial consortium reductively dechlor
150. PCB TOT BEDE OBE 14 E g 1x7 PCB 5 50 P lt 0 01 10 28 PRA 5 PCB 36 P lt 0 01 WHER AU E 56 PCB 56 AL PCB 3
151. PCR DGGE DNA ISOIL extraction kit Nippon Gene Tokyo Japan DNA PCR 168 rRNA DGGE Bacteria 16S rRNA 341F 518R Muyzer et al 1993 4 zeg 16S rRNA 344F 518R Casamayor et al 2002 PCR DGGE 2 pTBlue Perfectly Blunt Cloning Kits Novagen Madison WL USA PCR 16S rRNA its DNA ABI 3100 Applied Biosystems Foster CA USA DNA GENBTYX program Version 7 Software Development Co Tokyo Japan NCBI BLAST AB282339 to AB282545 DDBJ
152. 0 20 40 60 80 100 120 140 160 180 200 Incubation period days Fig 3 1 Changes in the total amount of PCB residues w w a in the amounts of acetate lactate and sulfate mmol 15 ml culture b and in the microbial population cells ml c in Kuridashi BS culture over 196 days of the incubation 44 100 oO Total residual PCBs w w J without addition with addition 0 20 40 60 80 100 120 140 160 180 200 Incubation period days Fig 3 2 Changes in the total amount of PCB residues w w in Kuridashi BS culture over 196 days of the incubation with or without additional acetate lactate electron donors with appropriate intervals Arrows show the time that the electron donors were added into the culture Vertical bars denote standard deviation 3 3 5 SF BS 196 Ho Hig 3 3 7 SF BS PCB 28 SEF BS 56 196 PCB
153. 170 C 30 C min COR 300 C 3 C min 300 C 10 PCB KC 300 400 PCB GC mass spectrometer GC GC MS GC 17A MS QP 5050 Shimadzu Kyoto Japan GC ECD 70 eV GC flame ionization detector GC GC FID GC 14B Shimadzu Kyoto Japan PCB GC ECD PCB
154. 9 PCB 63 2005 Table 1 7 PCB concentration in the environments Environments Concentration Waters 60 11000 pg l Sediments 39 630000 pg dry g Soils 825 ug dry kg Atmosphere 16 880 pg m Pisces 1500 550000 pg wet g Shellfishery 200 160000 pg wet g Birds 4800 22000 pg wet g Mammalians 0 11 30000 ug dry kg 14 POPs 2005 LOE SIRROZ BT HORE ARI ROE L PCB LT ERR ED DIIS 4 PCB E ex 24 EARR kV beL PCB PCB PCB
155. 9C 9C vec C OrC c eC CST 9c Sov Vc CVC CVC C9ECITVEIE ET TET vv rc c Compositions of congeners Fig 3 6 The residual amounts of the PCB congeners in the culture contain pellets TYK 1 compared with the sterilized culture Numbers indicated under the figure correspond to individual composition of PCB congeners 12 tT dao e dures 3n 1unoure penpisosq 51 TYK 1 PCB 51 0 PCB 76 9 25 9 PCB Fig 3 6 3 4 PCB PCB
156. MS AL 19 2 2 2 4 PCB PCB 1 100 ml PCB
157. PCB 71 4 3 6 PCB GB0 5 GB PCB GB0 5 AL 10 PCB Table 4 6 SF PCB 61 GB0 3
158. PS Dried PS DPS PS SMH LE HOE LK PS Ground PS GPS PS SL 05mm Burnt soil BS PS 550 C 24 BS BS 0 mm OFF FILE EOE LI 3 BS PS Table 3 2 3 2 3 PCB 2 PCB 56 5 PCB PS DPS GPS BS 35 GBS 3 121 C 30 min 77 A 100 ml
159. pgcerz EUB338mix Archaea ARCIIS FISH Table 4 4 Bacteria EUB338mix 4 X UO Archaea ARC915 FISH Fig 4 2 Fig 4 3 LAURA 6 3IBISESRZTTOT RISO MEY TALL EO GB0 05 10 10 Table 4 4 Effects of particle seize on the abundance ratio of Bacteria and Archaea Conditions L GB2 0 GBO 5 GBO0 05 Bacteria EUB388mix 35 596 66 2 44 5 61 9 Archaea ARC915 33 5 39 2 44 1 35 4 The percentage value was calculated compared with the total numbers of microorganisms in each optical area Abbreviations PS paddy soil L liquid GB glass beads The condition names were explained in materials and methods The experiments were performed more than triplicate 4 3 5 Bacteria Archaea 16S rRNA Ft DGGE
160. 70 25 34 7 0 84 9 10 8 544 12 1 72 64 25 35 236 4 3 4 832 10 0 74 5 22 6 41 234 2 0 8 88 6 7 9 70 1 32 3 74 95 245 4 3 9 83 4 4 10 6 63 3 15 5 FER 110 236 34 2 6 86 9 11 5 68 6 14 9 105 234 34 1 2 87 5 11 7 68 3 22 10 97 117 86 245 23 2356 4 2345 2 1 1 81 4 14 6 67 9 17 0 101 150 245 25 236 246 3 1 82 7 9 4 643 26 5 118 245 34 1 9 78 1 11 6 68 5 8 8 103 71 246 25 26 34 1 2 872 12 0 71 0 22 4 138 234 245 0 3 88 6 17 3 70 9 11 0 136 236 236 0 2 73 0 19 4 81 6 25 6 149 77 236 245 34 34 0 2 87 6 12 5 60 6 19 2 176 2346 236 0 2 77 8 15 0 63 9 24 0 Total 100 0 855 8 6 603 16 1 Abbreviations PS paddy soil BS burnt soil SD standard deviation The residual amounts are expressed as mol of the sterile control The values are the averages after 56 days of incubation for 15 times of the serial transfers during the maintenance of the anaerobic PCB degrading activity Significant difference in the residual mol of each PCB congener was compared between BS and PS cultures by an Aspin Welch test The symbols and denote the significant differences at less than 5 1 and 0 1 of probability respectively 3 3 3 PCB BS PCB Fig 3 1
161. LS 2006 Hee RXHZg P CHO DDT Dichloro diphenyl trichloro ethane PCB Polychlorinated bipheny VOR 12 POPs 2001 5 Polychlroinated dibenzo p dioxin PCDD Polychlorinated dib
162. PCB GB B 3 AL GBO 5 10 PCB PCB Yi gt eG AL 56 PCB 100 ug PCB 75 PCB
163. ET EDs PCB PCB Maks A JUR PCB Madigan et al EDS DIC WEE 7C PCB PCB B 2 FP D Hio ELI I SRI Sokol et al 1994 Wu et al 1996 2000 2002 Rysavy et al 2005 ya Clk Firmicutes
164. The scare was corresponded to squared distance Analysis was referred from http aoki2 si gunma u ac jp BlackBox BlackBox cgi GB0 5 PCR DGGE GB0 5 2 65 pmol 7 H2 0 02 nmol in the culture 7 H2 Firmicutes e 7zp7o7eo gc7e77g Epsilonproteobacteria Katayama and Fujie 2000 Bacteria Archaea PCR DGGE PS Fig 4 7 DGGE Firmicutes FA Deltaproteobacteria Table 4 5 PCB 70 Bacteria Archaea PS GBO 5 PS GBO 5 Fig 4 7 The DGGE
165. ug ml culture 0 064 ug atoms Cl g BS day 0 49 nmol C1 ml culture day PCB Quensen 1990 Hudson River PCB 3500 ug g PCB Aroclor 1242 1248 1254 1260 0 034 ug atoms Cl g sediment day Bedard 2006 300 pg ml Aroclor 1260 19 8 3 1 nmol Cl ml culture day 3 3 ug ml Aroclor 1260 0 16 0 03 nmol Cl ml culture day PCB BS ng total PCBs ml culture day Natarajan 19960 BS BS
166. 1 Composition of the gel solution for DGGE DGGE gel solution 40 Detergent AA solution 0 Formamide AA solution 6 5 ml 80 Formamide AA solution 6 5 ml APS 117 ul TEMED 11 7 pl 70 Detergent AA solution 0 Formamide AA solution 1 6 ml 80 Formamide AA solution 11 4 ml APS 117 ul TEMED 11 7 pl 0 Formamide AA solution 1 40 AA solution 250 ml 50xTAE 20 ml 80 acrylamide solution 1 40 AA solution 250 ml 50xTAE 20 ml Formamide 320 ml Urea 336 g 40 AA solution 1 50xTAE 10 ml Acrylamide 390 g Bis acrylamide 10g APS Ammonium pertoxodisulfate 0 1 w w TEMED N N N N tetramethylenediamine TAE 20 mM Tris 10 mM acetate 0 5 mM Na EDTA pH8 0 2 2 11 16S rRNA PCR DNA eg occoczges PCR 793F 5 ggg agt atc gac cct ctc tg 3 946R 5 cgt tye cct ttc rgt tca ct 3 Yoshida et al 2005 94 C 3 min 94 C 30 sec 55 C 30 sec 72 C 1 min 30 72 C 8 min PCR Dehal
167. 10 3 40 C PCB Sep Pack Plus Silica cartridges Waters Milford MA USA PCB 2 10 2 2 6 PCB
168. 2 2 5 5 CBP with BP PCB KC 200 300 400 PCB di CBP hexa CBP PCBs Abbreviations BP biphenyl CBP chlorinated biphenyl with indicated co metabolisms for degradation 12 Reference Ahmed and Focht 1972 Baxter et al 1975 Clark et al 1979 Focht and Brunner 1985 Furukawa et al 1978 Cookson 1995 Sierra et al 2003 Bedard et al 1983 Bedard et al 1990 1987 Unterman et al 1988 Komancova et al 2003 Albro and McKinney 1981 Furukawa et al 1983 Pettigrew et al 1990 Camandeur et al 1996 Kohler et al 1988 Mohn and Tiedje 1992 Dehalospirillium multivorans PCE Neumann et al 1998 Desulfitbacterium dehalogenans ozo Van de Pas et al 1999 Dehalococcoides ethenogenes TCE Magnuson et al 2000 Dehalococcoides Desulfitibacterium JE O Wut Nonaka et al 2006 Anaerobic
169. 2006 http www jesconet co jp MERL team 2002 Metals for Environmental Remediation and Learning CD R WHO 2006 Project for the re evaluation of human and mammalian toxic equivalency factors TEFs of dioxins and dioxin like compounds The International program on Chemical Safety 2005 14 POPs 2001 2005 14 FE POPs 2006 18 E 1999 PCB R727 KAR 2000 Beat Myki 2000 6 126 153 2006 REA 2006 2004 2006 Se Se Se XE ux F 91 5
170. 578 Quensen J F IIL Boyd S A Tiedje J M 1990 Dechlorination of four commercial polychlorinated biphenyl mixtures Alochlors by anaerobic microorganisms from sediments Appl Environ Microbiol 56 2360 2369 Sekiguchi Y Kamagata Y Nakamura K Ohashi A Harada H 1999 Fluorescenece in situ 88 hybridization using 16S rRNA trageted oligonucleotides reveals localization of methanogens and selected uncultured bacteria in mesophilic and thermophilic sludge granules Appl Microbiol Biotechnol 65 1280 1288 Shibata A Inoue Y Katayama A 2006 Aerobic and anaerobic biodegradation of phenol derivatives in various paddy soils Sci of Total Environ 367 979 987 Sinkkonen S Paasivirta J 2000 Degradation half life times of PCDDs PCDFs and PCBs for environmental fate modeling Chemosphere 40 943 949 Smits T H M Devenoges C Szynalski K Maillard J Holliger C 2004 Development of a real time PCR method for quantification of the three genera Dehalobacter Dehalococcoides and Desulfitobacterium in microbial communities J Microbiol Methods 57 369 378 Sokol R C Methoney C M Rhee G Y 1994 Effect of PCB concentration on reductive dechlorination and dechlorination potential in natural sediments Water Res 29 45 48 Someya T 1995 Three dimensional observation of soil bacteria in organic debris with a conforal laser scanning microscope Soil Microorg 46 61 69 Takasuga T Kumar K S N
171. A Fujie K 2000 Characterization of soil microbiota with quinone profile Soil biochem 10 303 347 Kim J Rhee G Y 1997 Population dynamics of polychlorinated biphenyl dechlorinating microorganisms in contaminated sediments Appl Environ Microbiol 63 1771 1776 Kim K S Hirai Y Kato M Urano K Masunaga S 2004 Detailed PCB congener patterns in incinerator flue gas and commercial PCB formulations Kanechlor Chemosphere 55 539 553 Krasotkina J Walters T Maruya K A Ragsdale S W 2001 Characterization of the Bi and iron sulfur containinc reductive dehalogenase from Desulfitobacterium chlororespirans J Biol Chem 276 40991 40997 Lee W Batchelor B 2004 Abiotic reductive dechlorination of chlorinated ethylenes by soil Chemsphere 55 705 713 L ffer F E Ritalahti K M Tiedje J M 1997 Dechlorination of chloroethenes is inhibited by 2 bromoethanesulfonate in the absence of methanogens Appl Environ Microbil 65 4982 4985 Magnuson J K Romine M E Burris D R Kingsley M T 2000 Trichloroethene reductive dehalogenase from Dehalococcoides ethenogenes sequence of tceA and substrate range Appl Environ Microbiol 66 5141 5147 Maym Gatell X Chien Y T Gossett J M Zinder S H 1997 Isolation of a bacterium that reductively dechlorinates tetrachloroethene to ethane Science 276 1568 1571 Meckenstock R U Annweiler E Michaelis W Richnow H H Schink B 2000
172. Dehalobacter E T PCB E PCB PCB cz7es 18 2 2 2 2 1 PCB PCB PCB PCB KC 300 KC 400 1 1 PCB 2 7 PCB 30 5 PCB 3 39 9 w w 4 44 4 w w PCB 1000 mg l 2 2 2
173. PCB PCB PCB Gas chromatography GC GC 3 PCB Ni electron capture detector GC GC ECD GC2014 Shimadzu Kyoto Japan 1 HT8 0 22 mm inner diameter 50 m length 0 25 um film thickness SEG Austin TX USA 0 1 ml min Grade 1 30 ml min Grade 2 250 C 330 C 80 C 3
174. PCB 100 PCB E ii PCB ug 15 ml culture 10 1 PCB PCB HiH tA PCB 46 FRA ER BE TILER 3 Bo METZ PCB PCB AL 5 36 34
175. and PS cultures by an Aspin Welch test The symbols and denote the significant differences at less than 5 1 and 0 1 of probability respectively 65 PS 6 17 32 6 18 3 BS 17 826 850 35 X 0832 10 46 66 70 74 10 3 4 BS GB0 5 PS PCB 3 4 BS PCB 4 3 4 FISH Archaea Bacteria
176. consortium Biotechnol Let 21 741 745 Natarajan M R Wang H R 1995 Effect of oxygen and strange condition on the metabolic activities of polychlorinated biphenyls dechlorinationg microbial granules Appl Microbiol Biotechnol 43 733 738 Nath K Das D 2004 Improvement of fermentative hydrogen production various approaches Appl Microbiol Biotechnol 65 520 529 Neumann A Wohlfarth G Diekert G 1998 Tetrachloroethene dehalogenase from Dehalospirillum multivorans cloning sequencing of the encoding genes and expression of the pceA gene in Escherichia coli J Bacteriol 180 4140 4145 Nonaka H Keresztes G Shinoda Y Ikenega Y Abe M Naito K Inatomi K Furukawa K Inui M Yukawa H 2006 Complete genome sequence of the dehalorespiring bacterium Desulfitobacterium hafinense Y51 and comparison with Dehalococcoides ethenogenes 195 J Bavteriol 188 2262 2274 Palekar L D Maruya K A Kostka J E Wiegel J 2003 Dehalogenation of 2 6 dibromobiphenyl and 2 3 4 5 6 pentachlorobiphenyls in contaminated estuarine sediment Chemosphere 53 593 600 Roser D J 1980 Ethidium bromide a general purpose fluorescent stain for nucleic acid in bacteria and eukaryotes and its use in microbial ecology studies Soil Biol Biochem 12 329 336 Rysavy J P Yan T Novak P J 2005 Enrichment of anaerobic polychlorinated biphenyl dechlorinators from sediment with iron as a hydrogen source Water Res 39 569
177. et al 2006 PS BS PCR DGGE 4Archaea A3 Bacteria Fig 3 5 PS Bacteria Firmicutes Deltaproteobacteria Chloroflexi BS PCB Chloroflexi Dehalococcoides o 17 DF 1 Dehalobacter Desulfitobacterium BS PCR PCR DGGE PCB
178. fragment patterns of the amplified 16S rRNA genes for Bacteria and Archaea from microcosms of the two cultures after 56 days of incubation Abbreviation PS and GBO 5 are corresponded to the paddy soil PS culture developed in chapter 2 and the glass beads 0 5 mm culture The bands numbered in circles were subjected to the DNA sequencing and the phylogenetic analysis as shown in Table 4 5 Table 4 5 Phylogenetic assignment of the DGGE clones of partial 16S rRNA genes isolated from the microcosm in the BS culture Band Phylum class Closestrelative Accession Similarity No 96 1 Gammaproteobacteria Pseudomonas sp SBW25 AY45712 100 2 Bacteroidetes Uncultured bacterium BTSE T2 1F AY217446 99 from TCE dechlorinating consortia 3 Firmicutes Acetanaeobacter thermotolerans AF358114 99 4 Firmicutes Clostridium glycolicum strain EIB4 AY458856 99 5 Firmicutes Sedimentbacter hongkongensis AY571338 98 strain KI 6 Chloroflexi Uncultured Chloroflexi bacterium AY921709 99 sp 7 Methanomicrobia Methanoculleus palmolei Y16382 99 The band numbers correspond to those shown in circle in Fig 4 7 15 GB0 5 16S rRNA ez ococczdes AURAL 15 3 El Desulfitobacterium XU X 10 El Dehalobacter RURAL 6
179. ml mE a 4 4 PCB 4 2002 PCB 4 PCB H E E PCB E PCB E Fennel et al 2004 Cutter et al 2001 Wu et al E PEUX PCB tA EP SUR C RO WAD TET STD MEEL PCB PCB ETZE LTR OE OKA WO
180. oZgc7e7 restrictus Holliger et al 1998 D multivorans Scholz Muramatsu et al 1995 Dehalococcoides ethenogenes Strain 195 Maymo Gatell 1997 Clostridium Methanococcous J amp Methanosarcina 11 Table 1 8 Aerobic PCB degrading bacteria Isolates Achromobacter Nocardia sp Pseudomonas sp Alcalegenes odorans A denitrificans Acinetobacter strain P6 Acinetobacter strain P6 Arthrobacter strain BIB Alcalegenes H850 Janibacter MS3 02 Alcalegenes eutrophus H850 Pseudomonas putida LB400 Alcalegenes eutrophus Pseudomonas putida Corynebacterium sp Pseudomonas sp 2 Pseudomonas testrosterone B356 Acinetobacter sp Achromobacter sp Bacillus brevis Alcaligenes sp JB1 Pseudomonas aeroginosa Pseudomonas putida metabolites BP 4 CBP PCB Aroclor1242 Aroclor1254 with BP BP 4 CBP Aroclor1254 with BP BP monoCBP Aroclor1242 1248 1254 Aroclor1242 2 3 5 6 position CBP 2 3 4 4 5 CBP 2 3 3 4 CBP 2 4 4 2 2 5 2 2 5 5 2 24 5
181. 1 Wu et al 2002 Natarjan et al 1998 Yan et al 2005 PS 16 37 180 199 4 GB0 5 FH PCB PCB PCB PCB 61 ll
182. 1 2 1 4 4 PCB o 17 Cutter et al 2001 Dehalobium chlorocoercia DF 1 DF 1 Wu et al 2002 Girmf RI EDS 2 3 5 6 TeCB Dehalococcoides ethenogenes strain 195 Fennell et al 2004 2 3 4 5 6 HCB Table 1 10 HE Yan Dehalococcoides eg opgc7e7 2 3 4 5 TeCB Yan et aL 2006 PCB PCB PCB Table 1 10 Anaerobic PCB dechlorinating bacteria Isolates Uncultured bacterium PCB Reference Dehalococcoides ethenogenes Strain 195 2 3 4 5 6 CBP Fennell et al 2004 DF 1 Dehalobium chlorocoercia DF 1 2 3 5 6 CBP Wu et al 2002
183. 10 ml 10 10 107 10 100 ug ml 100 90 PBS PBSE EDTA 1 1 50 10 Roser 1978 1995 3 2 12 DNA
184. 13 PCB 84 8 9 9 mol 85 7 10 6 Cw w 14 0 10 3 mol P lt 0 01 PS PCB 56 PS PCB mol Table 2 4 PCB PCB PCB PCB Teli S 26 50 41 B RUM6
185. 7 1 502 49 43 24 25 235 2 3 6 86 7 120 66 24 34 52 82 1 9 s 52 69 25 25 246 3 6 5 822 99 we 70 25 34 6 1 85 0 94 se 72 64 25 35 236 4 1 9 815 155 Al 234 2 0 6 85 7 40 6 74 95 245 4 236 25 4 5 759 98 sw 105 234 34 0 9 65 2 296 110 236 34 1 0 86 8 127 s 97 117 86 245 23 2356 4 2345 2 0 8 89 1 114 101 150 245 25 236 246 1 6 739 181 118 245 34 1 5 76 7 102 103 71 246 25 26 34 1 3 785 144 138 234 245 0 3 832 179 136 236 236 0 2 73 8 281 149 77 236 245 34 34 0 8 81 1 106 176 2346 236 0 1 55 77 Total 100 0 84 8 99 te SD standard deviation Significant differences in the residual mol of each PCB congener were examined by an Aspin Welch test The symbols and denote A significant difference at 5 1 and lt 0 1 respectively 29 2 3 3 PCB 56 PS 6 7 0 02 nmol culture PS PS 6 7
186. 81 343 1 46 23 26 4 7 67 6 1 9 62 7 6 8 78 4 4 3 49 43 24 25 235 2 3 2 67 8 2 8 111 4 6 6 82 143 4 66 24 34 5 9 64 5x3 0 75 8 4 8 79 0 4 2 52 69 25 25 246 3 5 4 68 6 2 6 66 8 4 3 81 1 3 6 70 25 34 7 1 65 4 2 5 71 7 7 2 77 245 4 72 64 25 35 236 4 3 0 66 2 2 6 82 9 2 1 83 0 3 2 41 234 2 0 8 65 9x2 5 83 0 7 4 83 7 3 1 74 95 245 4 3 7 63 343 3 78 4 5 6 77 5 3 8 110 236 34 0 9 62 2 0 7 74 8 S 1 78 9 3 7 105 234 34 1 1 64 6x1 3 79 0 4 6 76 2 3 1 97 117 86 245 23 2356 4 2345 2 0 2 67 1 1 3 89 841 5 80 0 2 5 101 150 245 25 236 246 3 4 65 5 0 2 81 447 5 77 7435 118 245 34 2 0 63 6 0 1 74 945 2 77 742 9 103 71 246 25 26 34 1 4 66 6 2 4 86 7 4 0 82 242 9 138 234 245 0 3 64 2 9 1 93 2 3 0 78 6 2 3 136 236 236 0 1 62 0 2 9 113 0 3 0 74 3 6 7 149 77 236 245 34 34 1 9 63 5x0 3 75 8 4 7 76 1 2 6 176 2346 236 0 1 65 0 0 7 85 345 5 77 4 3 2 Total 100 0 65 6 4 6 77 6 15 6 al 80 6 4 3 BES 2 bromoethane sulfonic acid Significant difference D in the residual mol percentage of each congener was examined by analysis of variance F test followed by a Bonferroni multiple analysis All results were compared with the BS culture after 56 days of incubation The symbols and denote the significant differences at less than 5 1 and 0 1 of probability respectively The proportions mol of two congeners 15 and 25 were not accurate because the contents were less than the detection limit in 100 ug 365 mmol of the Kanechlor 300 400 mixt
187. Anaerobic naphthalene degradation by a sulfate reducing enrichment culture Appl Environ Microbiol 66 2743 2747 Mohn W W Kennedy K 1992 Reductive dehalogenation of chlorophenols by Desulfomonile tiedjei DCB 1 Appl Enviorn Microbiol 58 1367 1370 Mohn W W Tiedje J M 1990 Catabolic thiosulfate disproportion action amd carbon dioxide reduction in strain DCB 1 a reductively dechlorinating anaerobe J Bacteriol 172 2065 2070 87 Mohn W W Tiedje J M 1991 Evidence for chemiosmotic coupling of reductive dechlorination and ATP synthesis in Desulfomonile tiedjei Arc Microbiol 157 1 6 Mohn W W Tiedje J M 1992 Microbial reductive dehalogenation Microbiol Rev 56 482 507 Muyzer G de Waal E C Uitterlinden A G 1993 Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction amplified genes coding for 16S rRNA Appl Environ Microbiol 59 695 700 Natarjan M R Wu W M Nye L W Wang H Bhatanager L Jain M K 1996 Dechlorination of polychlorinated biphenyl congeners by an anaerobic microbial consortium Appl Microbiol Biotechnol 46 673 677 Natarjan M R Wu W M Wang H Bhatnagar L Jain M K 1998 Dechlorination of spiked PCBs in lake sediment by anaerobic microbial granules Water Res 32 3013 3020 Natarajan M R Wu W M Sanford R Jain M K 1999 Degradation of biphenyl by methanogenic microbial
188. B 4 PCB PCB PCB 4 Firmicutes E HE Ch OS PS Chloroflexi P Deltaporteobacteria E BS HES GBO S DGGE BS Frimicutes Deltaporeteobacteria W C
189. B GB Dso 0 05 mm 0 5 mm 2 0 mm 3 EO GB Potters Ballotini Co Ltd Ibaraki Japan GB 10 HNO HNO 4 2 3 PCB PCB AL 56 5 2 3 GB 4 g 100 ml 121 C 30 min Coy Laboratory Products Grass Lake MD1 g l PCB 100 ul 2 AL 15 ml GB 100 GB AL GB G
190. B AL GB L GB GB0 5 GB2 0 GB0 05 PCB 5 Vy 30 C 56 PCB PCB 2 3 121 C 30 0 1 4 2 4 PS 2
191. DI Tolerable daily intake 4 pg TEQ kg day PCB PCB 2005 Tm Tm 1 3 2 PCB 1881 PCB PCB
192. IWA 3 TYK 1 B EX vi Table 3 6 Chemical and physical properties of pellets and the total PCB residues in the cultures after 56 days of incubation Pellet Specific surface area m7g Total pore capacity ml g Averaged pore diameter nm Porosity 96 Total residual PCBs 96 w w Total residual PCBs in sterilized sample 96 w w NIWA 1 NIWA 3 TYK 1 19 2 57 5 35 7 0 047 0 046 0 573 97 9 31 5 47 3 35 2 n d n d n t LS 51 0 n t 13 0 76 9 Abbreviation n d not detected n t not tested All data were analyzed by TYK co ltd except the total residual PCBs chemical properites were as follow 96 w w SiO 47 6 AbO 24 6 Fe O 1 8 TiO 1 0 MnO 0 1 Cr20 0 0 ZrO 0 0 CaO 10 7 MgO 1 93 Na O 1 2 K O 8 6 P20 8 7 SO 0 1 ZnO 0 1 Igloss 0 6 50 9 9 C E 5g bE PE SHT 9ET 9 8 9 c 9 c 2 2 B ctC EC oD x YE 97cC 9C E EE YE cc 9 C 9 SS C CSVECIV OSETIET evc Ve 9ET VE VET V SVC C VET V 9ET SE ST VE SC 9VUST ST ec c CSEC ST VT 9C SC
193. LL PS GB2 0 PCB GB0 05 PS PCB PS L 8 P lt 0 05 27 P lt 0 01 25 P lt 0 01 28 P lt 0 01 72 P lt 0 05 Nei 0 05 149 P lt 0 05 105 P lt 0 05 GB0O 5 PCB 27 P lt 0 01 25 P lt 0 01 GB2 0 GB0 05 PS 27 P lt 0 05 0 P lt 0 05 105 P lt 0 05 138 P lt 0 05 3 GB0 5 L GB2 0 5 GBO 05 PCB Fig 4 1 GB0 PCB GB0 L 2
194. OCDD 0 0003 PCDFs 2 3 7 8 TCDF 0 1 1 2 3 7 8 PeCDF 0 03 2 3 4 7 8 PeCDF 0 3 1 2 3 4 7 8 HxCDF 0 1 1 2 3 6 7 8 HxCDF 0 1 1 2 3 7 8 9 HxCDF 0 1 2 3 4 6 7 8 HxCDF 0 1 1 2 3 4 6 7 8 HpCDF 0 01 1 2 3 4 7 8 9 HpCDF 0 01 OCDF 0 0003 PCBs 3 4 4 5 TCB 81 0 0003 coplanar PCBs 3 3 4 4 TCB 77 0 0001 3 3 4 4 5 PeCB 126 0 1 3 3 4 4 5 5 HxCB 169 0 03 2 3 3 4 4 PeCB 105 0 0003 2 3 4 4 5 PeCB 114 0 0003 2 3 4 4 5 PeCB 118 0 0003 2 3 4 4 5 PeCB 123 0 0003 2 3 3 4 4 5 HxCB 156 0 0003 2 3 3 4 4 5 HxCB 157 0 0003 2 3 4 4 5 5 HxCB 167 0 0003 2 3 3 4 4 5 5 HpCB 189 0 0003 TEF were redefined by WHO 2006 KC300 KC400 N N N N N N N N N N N IN KC500 KC600 D 1CI E 2Cl 3CI amp 4CI B 5CI 6CI t3 7CI 8CI 9CI m 10CI Fig 1 3 PCB congener pattern percentage in each KC Kanechlor series HI 120 t PCB 1954 1972 58 787 t PCB 1 158 t PCB 54 001 t 2005 1968 PCB
195. OS DEB ou onde phu tata ut op tutta e nf ettet d 83 ME E 84 DI Pa C 85 1 1 1 A 10 100 2000 A
196. R ATES 1 LE E E ES PCB L PCB 1 5 3 PCB E PCB
197. a J Taleon D M Auresenia J Gallardo S 2005 Polychlotinated biphenyls and their biodegradation Process Biochem 40 1999 2013 Bouchard B Beaudet R Villemur R McSween G L pine F Bisaillon J G 1996 Isolation and characterization of Desulfitobacterium frappieri sp nov an anaerobic bacterium which reductively dechlorinates pentachlorophenol to 3 chlorophenol Int J Syst Bacteriol 46 1010 1015 Boyle A W Balke C K Prince W A II May H D 1993 Effects of polychlorinated biphenyl congener concentration and sediment supplementation on rates of methanogenesis and 2 3 6 trichlorobiphenyl dechlorination in anaerobic enrichment Appl Environ Microbiol 59 3027 3031 Brown J E Bedard D L Brennnan M J Carnahan J C Feng H Wager R E 1987 Polychlorinated biphenyl dechlorination in aquatic sediments Science 236 709 712 85 Casamayor E O Massana R Benlloch S vre s L D ez B Goddard V J Gasol J M Joint I Rodr guez Valera F Pedr s Ali C 2002 Changes in archaeal bacterial and eukaryal assemblages along a salinity gradient by comparison of genetic fingerprinting methods in a multipond solar saltern Environ Microbiol 4 338 348 Chang B V Liu W G 2001 Microbial dechlorination of three PCB congeners in river sediment Chemosphere 45 849 856 Choi S D Hong H B Chang Y S 2003 Adsorption of halogenated aromatic pollutants by a protein released
198. ading activity over 15 times of the serial transfers IUPAC Substituted positions of mol in initial GB 0 5 Significant number Cl PCB mixture Residual amount SD difference 6 2 3 0 6 104 7 112 9 8 2 4 4 5 38 8 15 9 16 23 2 0 3 63 1 188 20 33 45 23 3 34 2 236 2 7 9 57 6 15 9 17 24 2 2 8 50 0 145 28 53 24 4 25 26 10 1 51 2 160 18 25 2 7 7 70 3 404 26 50 25 3 246 2 1 3 55 9 146 31 25 4 9 6 58 8 166 32 26 4 2 0 55 2 23 8 44 23 25 6 1 71 3 192 46 23 26 3 2 72 2 168 49 43 24 25 235 2 4 2 64 8 22 1 66 24 34 5 8 70 5 110 52 69 25 25 246 3 6 6 79 0 22 7 70 25 34 7 0 69 8 6 72 64 25 35 236 4 3 4 65 7 183 41 234 2 0 8 69 9 234 74 95 245 4 3 9 73 8 139 110 236 34 2 6 83 9 6 6 105 234 34 1 2 78 8 17 3 97 117 86 245 23 2356 4 2345 2 1 1 75 2 187 101 150 245 25 236 246 3 1 76 1 12 0 118 245 34 1 9 76 3 9 7 103 71 246 25 26 34 1 2 69 2 160 138 234 245 0 3 83 6 16 2 136 236 236 0 2 705 268 149 77 236 245 34 34 0 2 79 7 143 176 2346 236 0 2 76 7 12 0 Total 100 0 58 4 12 0 GB glass beads SD standard deviation The residual amounts are expressed as mol of the sterile control The values are the averages after 56 days of incubation for 15 times of the serial transfers during the maintenance of the anaerobic PCB degrading activity Significant difference in the residual mol of each PCB congener was compared between GB
199. al PCB structures correspond to IUPAC congener numbers Symbols and on the congener numbers correspond to the results of a Bonferroni multiple comparison as shown in Table 3 4 Congeners 6 15 25 and 26 were accumulated congeners more than 100 mol while 28 and 44 with dotted line were significantly decreased congeners BS DGGE Firmicutes P Deltaproteobacteria Chloroflexi Fig 3 5 Deltaproteobacteria Desulfovibrio BS PCB PCB PCB PCB Bil A X Baltimore ozo Holloman et al 1998 Venice Lagoon meta para Fava et al 2003
200. bacterium Table 2 6 Dehalococcoides KE Dehalobacter RITIKA b VIO 3 Desulfitobacterium JY s EE 40 t TE Fig 2 1 PCR DGGE profiles of cultures during serial transfers Lanes 1 and 2 are the duplicated results of the consortium that developed after 13 serial transfers of the anaerobic PCB degrading culture obtained from Kuridashi paddy soil Lane O indicates the original paddy soil The percentages on the left hand side of the figure indicate the gradient percentage of the denaturant in this gel The bands numbered in circles were sequenced The phylogenetic 70 analysis of these bands is shown in Table 2 5 30 Table 2 5 Phylogenetic analysis of DGGE bands obtained from a Kuridashi paddy soil culture that developed after 13 serial transfers of the anaerobic PCB degrading culture Band Phylum class Closest relative ACCESSION Similarity No Uncultured bacterial clone Z39 from 1 Firmicutes Hudson river PCB dechlorination AY 154833 97 sediment METES Uncu
201. bromoethane sulfonic acid BES Gunsalus et al 1978 AL pH AL pH7 pHS pH9 AL 4 C 22 C 100 rpmmin 2 35 6 AL PS PCB PS 3 2 5 PCB 1 mmol BS SF BS H BS 14 28 H 56 112 196 36 3 2 6 BS PCB
202. crobiol Ecol 44 309 318 Fennell D E Nijenhuis L Wilson S E Zinder S H H ggblom M M 2004 Dehalococcoides ethenogenes strain 195 reductively dechlorinates diverse chlorinated aromatic pollutants Environ Sci Technol 38 2075 2081 Garbarini D R Lion L W 1986 Influence of the nature of soil organics on the sorption of tolene and trichloroethylene Environ Sci Technol 20 1263 1269 Gunsalus R P Romesser J A Wolfe R S 1978 Preparation of coenzyme M analogues and their activity in the metyl coenzyme M reductase system of Methanobacterium thermoautorophicum Biochem 17 2374 2377 86 Hartkamp commandeur L C M Gerritse J Govers H A J Parsons J R 1996 Reductive dehalogenation of polychlorinated biphenyls by anaerobic microorganisms enriched from duch sediments Chemosphere 32 1275 1286 Holliger C Hahn D Harmsen H Ludwig W Schumacher W Tindall B Vazquez F Weiss N Zhender A J B 1998 Dehalobacter restrictus gen nov and sp nov a strictly anaerobic bacterium that reductively dechlorinates tetra and trichloroethene in an anaerobic respiration Arch Microbiol 169 313 321 Holloman T R P Elberson M A Cutter L A May H D Sowers K R 1998 Characterization of a defined 2 3 4 5 tetrachlorobiphenyls ortho dechlorinating microbial community by comparative sequence analysis of genes coding for 16S rRNA Appl Environ Microbiol 64 3359 3367 Katayama
203. data not shown PS GB0 5 Table 4 1 Effects of particle size on the total residual PCBs in second transferred cultures from PS culture Conditions PS L GB2 0 GBO 5 GB0 05 Material Particle size Soil Non GB 2 0 GB 0 5 GB 0 05 Residual amount 86 0 42 6 89 2 8 3 77 3 11 6 72 9 6 8 92 5 42 0 weight Abbreviations PS paddy soil L liquid GB glass beads The condition names were explained in materials and methods The experiments were performed more than triplicate Significant decrease in the total residual weight was compared with PS culture by analysis of variance F test followed by a Scheffe multiple comparison test The symbol denote the significant differences at less than 5 of probability 4 3 2 PCB Table 4 2 PS GB0 5 2 PCB PCB GB0 5
204. dechlorination e donor Cl C Cl Ote O5 Cl Cl OA o Cl Cl Highly chlorinated biphenyls Fig 1 5 Anaerobic dechlorination of highly chlorinated biphenyls Cl 1 4 3 PCB PCB PCB Chang et al 2001 Fava et al 2003 PCB PCB PCB Mohn and Tiedje 1992 Wu et al 1998 Palkar et al 2003 Tiedje et al 1993 Borja et al 2005 Wiegel and Wu 2000 Table 1 9 PCB Brown 1987 PCB Bedard and Quensen 1995
205. e W solution 1 ml Resazurin solution 1 ml FeS solution 1 ml 200 mM MOPS solution 100 ml pH 7 by KOH Trace element SL 10 I HCl 25 10 ml FeCls 4H2O 1 5 g ZnCl 0 07 g MnCl 4H20 0 1 g H3BO 6 mg CoCls 4H2O 0 19 g CuCLl 2H 0 2 mg NiCl 6H O 0 024 g Na MoO 2H20 0 036 g Se W solution l NaOH 0 5 g NaSeO3 5H20 3 mg Na2WO4 2H 0 4 mg SF SF BS SF BS 5i 2 3 121 C 30 0 1 3 24 PS PS PCB PS 100 C 10 min 2 2 mM Taylor and Oremland 1979 2 mM 2
206. electron donors were added into the culture Vertical bars denote standard deviation 3 3 6 PCB BS PCB ry BS PCB 77 6415 6 mol BS PCB 65 6 4 6 mo1l 25 3 1 4 mol 34 543 9 mo1 17 3 mo1 B V PCB 328 453 P lt 0 01 44 P lt 0 01 BS S Table 3 4 PCB 15 P lt 0 01
207. enzofuran PCDFD PCB rh Hom 2000 POPs 200 2006 2003 2 AUS DUN Hig 1 1 POPs Persistent Organic Pollutants gt de JA B BE 10 E
208. er Lake Woods pond Baltimore Harbor Starch amp whey proceccing UASB River Rhine Chemie Harbour Germany Di Hua Mi Shi sewage plant Taiwan Keelung River Taiwan Tansui River Erjen River Taiwan Porto Marghera Venice Laggon Italy Abbreviation CBP chlorinated biphenyl mark indicated PCB uncontaminated microbial source Table 1 9 Anaerobic PCB dechlorinating activities and consortium PCBs Aroclor1242 1254 Aroclor1242 1254 Aroclor1260 1254 Aroclor1254 1260 2 3 4 5 6 2 3 5 6 2 3 4 5 CBP Aroclor1260 Aroclor1260 2 3 4 5 6 234 2 2 3 3 4 5 6 CBP 2 2 3 3 4 4 2 2 3 3 6 6 CBP 2 3 4 5 CBP and other 10 congeners 11 congeners Co planar PCBs 2 3 4 5 6 CBP Aroclor1242 1254 Main activities meta para meta para meta amp p ara 53 for 16 w eeks 76 for 16 weeks ortho 20 for 12 weeks 80 for 75 weeks Consortium BES inhibition o 17 DF 1 Dehaloco ccoides like bacteria References Alder et al 1993 Quensen et al 1990 Ye et al 1992 Sokol et al 1994 Quensen et al 1988 1990 Zwiernik et al 1998 May et al 1992 Bedard et al 1996 2005 2006 Quensen et al 1990 Berkaw et al 1996 Cutter et al 1998 amp 2001 Wu et al 1998 amp 2002 Yan et al 2006 Natarjan 1995 1996 amp 1998 Nollet 2005 Hartkamp commandeur et al 1996 Chang et al 1998 Chang et al 2001 Kuo et al 1999 Fava et al 2003
209. from Bacillus pumilus Water Res 33 4004 4010 Cutter L Sowers K S May H D 1998 Microbial dechlorination of 2 3 5 6 tetrachlorobiphenyl under anaerobic conditions in the absence of soil or sediment Appl Environ Microbiol 64 2966 2969 Cutter L A Watts J E M Sowers K R May H D 2001 Identification of a microorganism that links its growth to the reductive dechlorination of 2 3 4 5 chlorobiphenyl Environ Microbiol 3 699 709 Fagervold S K Watts J E M May H D Sowers K R 2006 Sequential reductive dechlorination of meta chlorinated polychlorinated biphenyl congeners in sediment microcosms by two different Chloroflexi phylotypes Appl Environ Microbiol 71 8085 8090 Fantroussi S E Naveau H Afathos S N 1998 Anaerobic dechlorinating bacteria Biotechnol Prog 14 167 188 Fava F Gentilucci S Zanaroli G 2003 Anaerobic biodegradation of weathered polychlorinated biphenyls PCBs in contaminated sediments of Porto Margera Venice Lagoon Italy Chemosphere 53 101 109 Fava E Gioia D Marchetti L Quattroni G 1996 Aerobic dechlorination of low chlorinated biphenyls by bacterial biofilms in packed bed batch bioreactors Appl Microbiol Biotechnol 45 562 568 Fava F Zanaroli G Young L Y 2003 Microbial reductive dechlorination of pre existing PCBs and spiked 2 3 4 5 6 pentachlorobiphenyls in anaerobic slurries of a contaminated sediment of Venice lagoon Italy FEMS Mi
210. hloroflexi GB0 5 Deltaproteobacteria Chloroflexi PJO DGGE PCR Chloroflexi PCB T AL BS Hee wit ME SLE PC 3 GB0 PCB AL PCB AL
211. il 55 5 29 5 50 5 18 9 CT mg kg dry soil 36 8 4 0 9 0 6 0 SO mg kg dry soil 92 1 36 2 216 2 60 3 pH H 0 6 7 6 3 6 4 6 2 Electric conductivity LS cm 50 1 37 0 63 7 21 5 Oxidation reduction potential mV 179 79 67 92 Total residual PCBs w w 85 6 8 6 77 6 9 4 78 0 9 1 77 5 7 2 PS paddy soil BS burnt soil ND less than detection limit 0 01 The exchangeable cations were evaluated by acetate ammonium extraction Analysis condition soil water 10 g dry weight 125 ml at 25 C Total PCB residues in individual cultures containing PS or BS are shown as percentage w w of the sterile control The values are the averages of three times of incubation using five percent v v of the cultures transfer with incubation intervals of 56 days Significant differences in the total PCB residues in BS cultures were compared with PS culture by analysis of variance F test followed by a Bonferroni multiple comparison test The symbol denote the significant differences at less than 5 of probability BS PCB 2 5 36 3 15 BS PCB
212. inating 1 2 3 and 1 2 4 trichlorobenzene Appl Enviorn Microbiol 64 496 503 Adrian L Szewzyk U Wecke J G risch H 2000 Bacterial dehalorespiration with chlorinated benzenes Nature 408 580 583 Ahmed M Focht D D 1973 Oxidation of polychlorinated biphenyls by achromobacter PCB Bull Environ Contam Toxicol 10 70 72 Bedard D L Bailey J J Reiss B L Jerzak G V S 2006 Development and characterization of stable sediment free anaerobic bacterial enrichment cultures that dechlorinate Aroclor 1260 Appl Environ Microbiol 72 2460 2470 Bedard D L Dort H V Deweerd K A 1998 Brominated biphenyls prime extensive microbial reductive dehalogenation of Aroclor 1260 in Housatonic river sediment Appl Environ Microbiol 64 1786 1795 Bedard D L Quensen J F III 1995 Microbial reductive dechlorination of polychlorinated biphenyls In Microbial transformation and degradation of toxic organic chemicals Young L Y and Cerniglia Eds Wiey Liss Division John Wiley and Sons New York 127 216 Berkaw M Sowers K R May H D 1996 Anaerobic ortho dechlorination of polychlorinated biphenyls by estuarine sediments from Baltimore Harbor Appl Environ Microbiol 62 2534 2539 Becker J G Berardesco G Rittmann B E Stahl D A 2005 The role of syntrophic associations in sustaining anaerobic mineralization of chlorinated organic compounds Environ Health Perspectives 113 310 316 Borj
213. iphenyls FEMS Micro Ecol 32 1 15 Wiegel J Zhang X Wu Q 1999 Anaerobic dehalogenation of hydroxylated polychlorinated 89 biphenyls by Desulfitobacterium dehalogenans Appl Environ Microbil 65 2217 2221 Wu Q Watts J E M Sowers K R May H D 2002 Identification of a bacterium that specifically catalyzes the reductive dechlorination of polychlorinated biphenyls with doubly flanked chlorines Appl Envrion Microbiol 68 807 812 Wu Q Bedard D L Wiegel J 1997 Temperature determines the pattern of anaerobic microbial dechlorination of Arochlor 1260 primed by 2 3 4 6 tetrachlorobiphenyl in Woods Pond sediment Appl Environ Microbiol 63 4818 4825 Wu Q Milliken C E Meier G P Watts E M Sowers K R May H D 2002 Dechlorination of chlorobenzenes by a culture containing bacterium DF 1 a PCB dechlorinating microorganism Environ Sci Technol 36 3290 3294 Wu Q Sowers K R May H D 1998 Microbial reductive dechlorination of Aroclor 1260 in anaerobic slurries of esturine sediments Appl Microbiol Environ 64 1052 1058 Wu Q Sowers K R May H D 2000 Establishment of polychlorinated biphenyl dechlorinating microbial consortium specific for double flanked chlorines in a defined sediment free medium Appl Environ Microbiol 66 49 53 Yan T LaPara T M Novak P J 2006 The effect of varying levels of sodium bicarbonate on polychlorinated biphenyl dechlorination in Hudso
214. ltured Clostridium sp clone 2 Firmicutes AC036 AY330126 100 3 Firmicutes Uncultured Clostridiales bacterium AB100470 95 4 Firmicutes Uncultured bacterial clone An7 AY826218 99 5 Firmicutes Uncultured bacterium AJ229190 95 6 Firmicutes Clostridium putrefaciens strain MH47 AJ318906 93 Uncultured bacterial clone TSAa22 7 Bacteroidetes from polychlorinated AB186816 99 dioxin dechlorinating community Delta Uncultured bacterium closely related to 8 proteobacteria Desulfovibrio desulfuricans G20 60240802 2n 9 Chloraea Uncultured Chloroflexi bacterial clone AY921709 99 AKYG643 The band numbers correspond to those shown in circles in Fig 2 1 Table 2 6 The frequency of detection of dechlorinating bacteria by specific PCR in Kuridashi paddy soil cultures during 13 serial transfers Dechlorinating bacteria Frequency of detection Dehalococcoides spp 0 13 Dehalobacter spp 3 13 Desulfitobacterium spp 10 13 2 4 KC 300 400 PCB 6 4 PCB 20
215. n River sediment cultures Environ Microbiol 8 1288 1298 Ye D Quensen J F III Tiedje J M Boyd S A 1995 Evidence for para dechlorination of polychlorobiphenyls by methanogenic bacteria Appl Environ Microbiol 61 2166 2171 Ye D Quensen J F IIL Tiedje J M Boyd S A 1999 2 Bromoethanesulfate sulfate molybate and ethanesulfonate inhibit anaerobic dechlorination of polychlorobiphenyls by pasterised microorganisms Appl Environ Microbiol 65 327 329 Yoshida N Takahashi N Hiraishi A 2005 Phylogenetic characterization of a polychlorinated dioxin dechlorinating microbial community by use of microcosm studies Appl Environ Microbiol 71 4325 4334 Yoshida N Yoshida Y Handa Y Kim H K Ichihara S Katayama A submitted Polyphasic characterization of an anaerobic microbial community completely dechlorinating pentachlorophenol to phenol enriched from paddy soil Zhang X Young L Y 1997 Carboxylation as an initial reaction in the anaerobic metabolism of naphthalene and phenanthrene by sulfidogenic consortia Appl Enviorn Microbiol 63 4759 4764 Zheng D Angenent L T Raskin L 2006 Monitoring granule formation in anaerobic upflow bioreactors using oligonucleotide hybridization probes Biotechnol Bioeng 94 458 472 Zwiernik M J Quensen III J E Boyd S A 1998 FeSO amendments stimulate extensive anaerobic PCB dechlorination Environ Sci Technol 32 3360 3365 90 JESCO
216. ncultured Chloroflexi bacterium sp 7 Methanobacteria Uncultured Methanobacterium sp 8 Methanomicrobia Methanoculleus palmolei The band numbers correspond to those shown in circle in Fig 3 5 49 Accession No AY754833 AJ488093 AY458856 AY 827462 AF418179 AY921709 DQ507195 Y16382 Similarity 97 100 100 96 99 99 98 99 3 3 8 RE NN RI ate Fig 3 5 ea dd V Wy Clk Firmicutes F4 Deltaproteobacteria Wd Chloroflexi 4 6 BS HED J 15 LI Jj Archaea Firmicutes FH1 BS 4 BS lt 3 BES DGGE
217. o 17 2 3 5 6 CBP Cutter et al 2001 Abbreviations CBP chlorinated biphenyl with indicated co metabolisms for degradation 15 1 5 1 PCB PCB PCB PCB PCB KC 300 400 PCB PCB PCB
218. obacter JE UTR FER U Desulfitobacterium 25 Smits ez opgc7er 441F 5 gtt agg gaa gaa cgg cat ctg t 3 645R 5 cct ctc ctg tcc tca age cat a 3 Desulfitobacterium 406F 5S gta cga cga agg cct teg get 3 619R 5 cce agg gtt gag ccc tag gt 3 2 2 12 DNA DDBJ DNA data bank of Japan AB259666 AB259674 2 3 2 3 1 PCB Table 2 2 4 6 3 PCB 4 PCB 6
219. of the amplified 16S rRNA genes for Bacteria from microcosms of serial transferred cultures for 5 types of medium after 56 days of incubation The numbers in circle corresponded to the times of serial transfer on individual cultures Five types of medium were abbreviated as PS L GB2 0 GBO 5 and GBO 05 which are described in materials and methods of this chapter detail The band patterns in individual cultures were analyzed by cluster analysis and indicated on Fig 4 4 PS L 2 00 5 0 05 PS L 2 0 0 5 0 05 iliis fat es i Fig 4 5 The DGGE fragment patterns of the amplified 16S rRNA genes for Archaea on serial transferred cultures for 5 types of medium after 56 days of incubation The numbers in circle corresponded to the times of serial transfer on individual cultures Five types of medium were abbreviated as PS L GB2 0 GBO 5 and GB0 05 which are described in materials and methods of this chapter detail 69 PS 1 PS 2 PS 3 PS 4 L 1 GB0 05 1 GB0 05 4 GB0 05 2 GB0 05 3 GB0 5 1 GB2 0 1 GB0 5 4 GB0 5 3 GB2 0 3 GB2 0 4 GB0 5 2 GB2 0 2 L 4 L 3 L 2 T B Ro Mee e d 0 100 200 300 Fig 4 6 Dendrogram from the cluster analysis grouping the DGGE samples of soil free cultures on serial transferred cultures from PS culture using Ward method normalized data The numbers with the name of the individual cultures were corresponded to the times of serial transfer
220. oma Y Sakai S 2005 Chemical characterization of polychlorinated biphenyls dibenzo p dioxins and dibenzofurans in technical Kanechlor PCB formations in Japan Arch Environ Contam Tocicol 49 385 395 Taylor B Oremland R S 1979 Depletion of adenosine triphospate in Desulfovibrio by oxyanions of group VI elements Current Microbiol 3 101 103 Tiedje J M Quensen J F III Sanford J C Schimel J P Boyd S A 1993 Microbial reductive dechlorination of PCBs Biodegradation 4 231 240 Utkin L Dalton D D Wiegel J 1995 Specifity of reductive dehalogenation of substituted ortho chlorophenols by Desulfitobacterium dehalogenans JW IU DC1 Appl Enviorn Microbiol 61 346 351 Van de Pas B A Smidt H Hagen W R vander Oost J Schraa G Stams A J de Vos W M 1999 Purification and molecular characterization of ortho chlorophenol reductive dehalogenase a key enzyme of halorespiration in Desulfitobacterium dehalogenans J Biol Chem 274 20287 20292 Voice T C Weber W J 1983 Sorption of hydrophobic compounds by sediments soils and suspended solids I Water Res 10 1433 1441 Watts J E M Fagervold S May H D Sowers K R 2005 A PCR based specific assay reveals a population of bacteria within the Chloroflexi associated with the reductive dehalogeneation of polychlorinated biphenyls Microbiology 151 2039 2046 Wiegel J Wu Q 2000 Microbial reductive dehalogenation of polychlorinated b
221. s PCB nt n gt aan 57 E PCB PCB EZZ E Cn 4 PCB 4 1 3 PCB
222. ted biphenyls Fig 1 4 Aerobic degradation of less chlorinated biphenyls 1 4 2 Halorespiraiion PCB ATP fig 1 5 3 Desulfomonile tiedjei Mohn and Tiedje 1990 1991 Desulfitobacterium dehalogenans van de Pas et al 1998 D chlororespirans LOffer et al 1996 PCE Tetrachloroethene e z
223. upplemented with MS or AL medium after four or six months of incubation Total Kuridashi 4 months 6 months MS medium Glucose 95 6 83 2 Lactate 94 4 82 6 Pyruvate 98 4 81 7 Acetate 97 5 80 6 Propionate 98 0 80 0 Ethanol 98 7 81 6 Nitrate 99 0 74 9 Mn IV 98 2 81 8 Fe IID 100 0 82 8 Sulfate 102 3 81 5 None 98 3 83 2 AL medium 89 2 TZS MS medium mineral salt medium AL medium acetate and lactate medium residual PCBs w w Nagakute 4 months 96 7 87 4 87 7 88 3 90 1 95 6 98 2 98 7 96 6 100 7 89 7 95 3 Yatomi 4 months 92 8 88 2 91 0 96 0 95 5 97 2 see text for the composition not tested Total residual PCBs are shown as 96 w w of the amount in the sterilized control after the same incubation time The values were the average of between two and seven determinations using separate incubation vials Statistical significance of decreases in the total residual PCBs compared with sterile control was examined by analysis of variance F test followed by a Scheffe multiple comparison The symbols and denote a significant difference at 5 1 and 0 1 respectively 2 3 2 PCB AL PS Table 2 3 56
224. ure used in this study BS BES BES PCB 80 6 4 3 mol 20 7 4 2 mol Table 3 4 Fig 3 4 BS PCB BES E Before incubation O Sterile control BS culture Molybdate Residual amount ug sample z Z z A be A 78 A Zt A5 A A Be H zz z M A 7 he AR m Z8 A Bares eo ON o oO oO y x 9 g E g 2 d 8 a a C1 N uv Congeners m Fig 3 4 The residual amounts of the selected PCB congeners in the individual incubation conditions The congeners significantly affected by inhibitors and the indicator congeners with under line of Kanechlor 300 400 Numbers indicated under individual PCB structures correspond to IUPAC congener numbers 3 3 7 BS
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