Khảo sát vi sinh vật tại vùng bị ảnh hưởng dioxin và đánh giá tiềm năng tiêu độc của vi sinh vật bản địa : Đề tài NCKH. QMT.07.02

DAI HOC QUOC GIA HA NOI 'kieic'k'k'kie'k'k'k'k BAG CAO KET QUA T H V C HIEN N H I E M V U K H O A HOC CONG NGHE PHUC VU QUAN LY NHA NUdC VE BAO VE MOI TRUCJNG Ten de tai: Khdo sat vi sink vat tai viing bi anh hu&ng dioxin vd ddnh gid tiem ndng tieu doc cua vi sink vat ban dia Ma so: QMT.07.02 Chu nhiem dex tai: TS Duong Van Hop Ca quan thyc hien: Vien Vi sinh vat va Cong nghe Sinh hoc, DAI HOC QUOC GIA HA NQI Ha ngiy thdng nam 2010 BAO CAO KET QUA THlTC HIEN NHIEM VU KHOA HOC CONG NGHE PHUC VU QUAN LY NHA NlTdC VE BAO VE MOI TRU'OHVG BAO CAO TOM TAT A Ten de tai Khao sat vi sinh vat tai viing hi dnh huomg dioxin va danh gid tiem ndng tieu doc cua vi sinh vat ban dia Ma s6 QMT 07.02 Thoi gian thyc hien nam (2007 - 2009) Cap quan ly Dai hoc Quoc gia Ha Noi Chu nhiem de tai TS Duong Van Hop Vien Vi sinh vat va Cong nghe sinh hoc (IMBT), DHQGHN Can bp tham gia chinh TS Dinh Thuy Hang ThS Nguyen Minh Giang ThS Phan Thi Ha TS Do Ngoc Lanh (IMBT) (IMBT) (IMBT) (Trung tam nhiet doi Viet-Nga) ThS Nguyen Thu Hoai (Trung tam nhiet dai Viet-Nga) Muc tieu va noi dung nghien ciru Muc tieu: - Phan tich vi sinh vat khu vuc nhiem dioxin nham tim cac nhom da c6 co hoi thich nghi vai chat doc va dong vai tro quan trpng qua trinh tay doc tir nhien tai noi nhilm - Danh gia tiem nang phan huy cua vi sinh vat ban dja thong qua phan tich boat tinh phan giai PAH va gen chuc nang dioxygenase Noi dung: - Thu thap mau dat va mau tram tich thuy virc tai khu nhiem doc san bay Da nlng va phan tich cac yeu to sinh-dia-hoa - Nghien CLTU da dang vi sinh vat c6 kha nang nuoi cay dugc phong thi nghiem thong qua phuong phap pha loang va nuoi cay tren moi truong thach hoac moi truong dich th^ (phuong phap MPN) - Nghien curu da dang vi sinh vat tong so (bao gom chua nuoi cay dirge va da nuoi cay dugc) b4no phuong phap PCR-DGGE doan gen ma cho 16S rDNA - Phan lap va nghien cuu cac chung vi sinh vat dai dien cho cac nhom c6 mat mau d^t/bun nhiem ve dac diem phan loai va hoat tinh sinh hoc - Nghien curu muc da dang cua cac gen chuc nang ma hoa cho dioxygenase thong qua phuong phap thi^t lap ngan hang gen Xac djnh cac gen cac chung vi sinh vat dai dien da phan lap dugc Cac kit qua dat diryc - D5 lira chgn dugc di^m thu mlu c6 muc nhiem doc tir thap den cao hai khu vuc nhiem doc dai dien, khu dat thugc bai luu tru dioxin chien tranh va ho sen chua nuac san bay Da Ning Mlu d^t va bun tai cac diem thu mau dugc thu thap va phan lop timg 20 cm va bao quan tai 4°C - Muc nhiem doc va cac yeu to sinh dia hoa tai diem thu mau da dugc phan tich, lam ca so de tien hanh khao sat vi sinh vat tai moi truong nghien cuu - Da tien hanh danh gia mat vi sinh vat tai diem thu mau doi vai cac nhom vi sinh vat hieu (bao gom vi khuan, xa khuan, nam sgi, nam men) va vi khuan ky (bao gom nhom khu nitrate, khu sulfate, khu Felll) Dong thoi ty le vi sinh vat c6 kha nang sinh truong vai carbazol cung diroc xac dinh - Tinh da dang cua vi khuan cac mau nhiem doc da dugc phan tich bang phuong phap FISH vai cac dau huynh quang dac hieu cho a-, p- va y-Proteobacteria - Da phan lap dugc 16 chung vi khuan va 44 chung nam sgi dai dien tai cac cac diem chpn lira va nghien cuu cac dac diem phan loai cung nhu hoat tinh sinh hoc cua chung Tinh da dang cua cac chung vi khuan dugc danh gia bang phuong phap ARDRA, tren ca sa da xac dinh dugc cac chi chiem uu the moi truong nhiem doc Cac chung nam sgi dugc phan loai dira tren cac dac diem hinh thai, tir danh gia tinh da dang va xac dinh cac chi chiem uu the moi truong nhiem doc - Trong so cac chung phan lap dugc da lira chgn chung vi khuan hieu khi, chung nam sgi va chung vi khuan ky khu nitrate the hien kha nang phan giai cao doi vai cac hop chat PAH, gom CO carbazol, naphthalene va phenanthrene - Da thiet lap dugc to hop vi sinh vat c6 hoat tinh phan huy cao doi vai cac hgp chat tham da nhan su dung thay the dioxin phong thi nghiem (carbazol, phenanthrene, naphthalene) C4u true quan the cua cac to hgp vi sinh vat dugc xac dinh bang phuong phap PCR-DGGE doi vai doan gen 16S rDNA - Dao tao hoc vien cao hoc - Cong bo bai bao khoa hgc Du kien tiep tuc cong bo them bai bao Tinh hinh kinh phi - T6ng kinh phi cua de tai: - Thanh toan dich vu cong cgng: - Chi phi thue muon: - Chi phi nghiep vu chuyen mon: 450.000.000 VND 37.250.000 215.000.000 243.750.000 Chu tri de tai ca quan EN TRUONG Xac nhan cua Dai hoc Quoc gia SUMMARY Project title: Microbiological survey of dioxin affected areas and assess the detoxifying potential of indigenous microorganisms Code: QMT.07.02 Duration: years (2007 - 2009) Organizer: Vietnam National University Hanoi (VNUH) Project leader: Dr Duong Van Hop Institute of Microbiology and Biotechnology (IMBT) - VNUH Key participants: Dr Dinh Thuy Hang (IMBT) M.Sc Nguyen Minh Giang (IMBT) M.Sc Phan Thi Ha (IMBT) Dr Do Ngoc Lanh (Vietnam-Russia Tropical Center) M.Sc Nguyen Thu Hoai (Vietnam-Russia Tropical Center) Objectives and study contents Objectives: - Analyse microorganisms in dioxin-contaminated areas in order to find out microbial groups that have been adapted to the toxic chemicals and that play important roles in detoxification processes in situ y^ca^^'^t - Access the degradation"potention\)f native microorganisms via analyzing degradation activity and genes coding for dioxygenases of the microorganisms Study contents: - Collect soil and mud samples at Danang airport and analyse biogeochemical characteristics of the samples - Study diversity of culturable microorganisms via methods of dilution and cultivation on solid or liquid media (MPN method) - Study diversity of total microorganisms (including culturable and unculturable) by using PCRDGGE method applied for 16S rDNA - Isolate and study phylogeny and biological properties of microbial strains that represent for the microbial groups existing in contaminated soil and mud samples - Study diversity of functional genes coding for dioxygenase via gene libraries Detect these genes in the isolated microbial strains The obtained results - Six sampling points with different dioxin contamination levels ranging from little to heavy were selected from two contaminated areas in Danang airport, including the former chemical storage area and the lotus pond Soil and mud samples at every sampling point were collected following the depth (to 80 cm) and every 20 cm of the samples were separately stored at 4°C - Contamination levels and biogeochemical factors of the collected samples were analysed in order to provide basis for microbiological analyses - Density of aerobic (bacteria, filamentous fiingi, yeasts) and anaerobic (nitrate reducers, iron reducers, sulfate reducers) microorganisms at the selected sampling points were identified At the same time, the proportion of aerobic carbazol-utilizing cells was also determined - Representatively, 16 bacterial strains and 44 fungous strains were isolated and studied on their phylogenetic characteristics and biological properties Diversity of the bacterial isolates was studied via ARDRA analyses, based on that the dominant groups at the contaminated sites were identified Fungous strains were classified based on phenotypic characteristics, their diversity and dominant groups in the contaminated sites were identified - Among the isolates, aerobic bacterial strains, fungous strains and nitrate-reducing bacterial strains were selected as representatives with high degradation capacity toward PAH compounds, including carbazol, naphthalene and phenanthrene - Succesflilly set up three consortia of microorganisms with high degradation capacity toward PAHs (carbazol, phenanthrene and naphthalene) that have been used in the laboratory as model compounds instead of dioxins Community structures of these consortia were analyzed by using PCR-DGGE method applied for 16S rDNA - Three postgraduated students have been working in this project - The project results have been published in three scientific papers In addition, one paper is planned to be published in the coming time ca quan Chu tri de tai PHO VIEN TRLTONG ac nhan cua Dai hoc Quoc g1a MVC LVC MdDAU CHirONG I T N G I QUAN TAI LIEU 1.1 Dioxin va hydrocarbon thom da nhan (PAH) 1.1.1 Giai thieu chung ve dioxin 1.1.2 Giai thieu chung ve PAH 1.2 Tinh hinh o nhiem dioxin va PAH tai Viet Nam va tren the giai 1.2.1 Tinh hinh nhiem dioxin 1.2.2 Tinh hinh nhiem PAH 1.3 Vai tro cua phan hiiy sinh hoc xir ly nhiem dioxin va PAH 1.3.1 Vi sinh vat phan giai dioxin 1.3.2 Vi sinh vat phan giai PAH 1.3.3 Cac nghien cuu phan buy sinh hgc dioxin va PAH a Viet Nam 1.4 San bay Da nIng - diem nong ve nhiem dioxin tai Viet nam 2 3 6 11 12 CHU*ONG II VAT LIEU VA PHLTONG PHAP NGHIEN ClTU 2.1 Vat lieu 2.1.1 Nguon mau va vi sinh vat 2.1.2 Hoa chdt, thiet bi 2.2 Cac phu'ong phap vi sinh vat 2.2.1 Xac dinh so lugng vi sinh vat hieu khi/ky 2.2.2 Lam giau vi sinh vat vai co chat PAH 2.2.3 Phan lap vi sinh vat hieu khiVky 2.2.4 Chup anh te bao dual kinh hien vi quang hgc 2.2.5 Xac dinh duang cong sinh truong thong qua protein tong so 2.3 Cac phu-ong phap phan tich hoa hoc 16 16 16 16 16 16 17 17 17 18 2.3.1 Phuong phap xac dinh S04^~ 18 2.3.2 Phuong phap xac djnh NOs" 19 2.3.3 Phuong phap xac dinh ham lugng mun 2.3.4 Phuong phap xac dinh nita tong so 2.3.5 Phuong phap xac dinh phosphor 2.3.6 Phuong phap xac djnh Fe tong so 2.3.7 Xac djnh PAH bang phuong phap hap phu UV-Vis 2.4 Cac phvong phap sinh hoc phan tir 2.4.1 Tach DNA tong so tu vi sinh vat 2.4.2 Phuong phap PCR 2.4.3 Dien di tren gel agarose 20 20 21 21 22 22 22 23 24 2.4.4 Dien di bien tinh (DGGE) 2.4.5 Phuong phap ARDRA 2.4.6 Phuong phap FISH 2.4.7 Phan tich da dang gen thong qua thilt lap thu vien gen 2.4.8 Giai trinh tir va dung cay phan loai 24 24 24 26 27 CHU'ONG HI KET QUA VA THAO LUAN 3.1 Thu mau nhiem dioxin va phan tich cac dac diem moi trvoug 3.2 Phan tich vi sinh vat 3.2.1 Xac dinh so lugng vi sinh vat cac mau nhiem doc 3.2.2 Phan lap cac chung vi sinh vat dai dien va nghien cuu tinh da dang cua chung 3.2.3 Nghien cuu da dang vi khuan cac mau nhiem doc bang phuong phap FISH 3.3 Nghien ciru kha nang phan huy PAH cua vi sinh vat tur vung nhiem doc 3.3.1 Tao cac quan the vi sinh vat vai kha nang phan giai PAH cao tu mau nhiem doc 3.3.2 Phan lap cac chung vi sinh vat phan giai PAH 3.4 Xac dinh gen dioxygenase quan the va cac chung da phan lap 3.5 Xac djnh gen dechlorinase quan the Ket luan Tai lieu tham khao Phu luc - Phu luc 1: Thanh phan moi truong nuoi cay vi sinh vat da sir dung - Phu luc 2: Cac trinh tu gen Phu luc 3: Cac cong trinh khoa hgc da cong bo Phu luc 4: Cac hgc vien cao hgc da tham gia dao tao Cac tai lieu khac 29 32 32 35 39 41 41 47 52 54 54 55 67 DANH MVC CAC C H f VIET TAT Fluorescence probe against a-Proteobacteria ALF968 ARDRA Amplified ribosomal DNA restriction analyses BET42a Fluorescence probe against ^-Proteobacteria CAR Carbazol CI CTAB DAPI DNA dNTP ddNTP DBT DD DF DGGE DMSO EDTA FISH FN GAM42a MPN MQ NTL PAH PCI PCR POP OD rDNA SDS TAE TBE 2,3,7,8-TCDD Chloroform - isoamylalcohol Cetyl trimethyl ammonium bromide 4',6-diamino-2-phenylindole Acid deoxyribonucleic Deoxyribonucleotide triphosphate Dideoxyribonucleotide triphosphate Dibenzothiophen Dibenzo-p-dioxin Dibenzofuran Denaturing gradient gel electrophoresis Dimethyl sulfoxide Ethylene-diamine-tetraacetic acid Fluorescence in situ hybridization Fluorene Fluorescence probe against y-Proteobacteria Most probable number Milli Q water Naphthalene Polycyclic aromatic hydrocarbon Phenol-chloroform-isoamylalcohol Polymerase chain reaction Persistent organic pollutants Optical density Ribosomal DNA Sodium dodecyl sulfate Tris/Acetate/EDTA Tris/Borate/EDTA 2,3,7,8-tetrachlorodibenzo-p-dioxin MODAU O nhiem dioxin, cu the la anh huang cua chat doc toi nguai va tim kiem bien phap tay doc hien dang la moi quan tam chung cua toan xa hoi Day la van de cap bach, nan giai va cung lai la van de nhay cam, lien quan den viec khac phuc hau qua sau chien tranh De co the xay dung dugc mot qui trinh cong nghe tly doc co hieu qua, cac nha khoa hgc can tien hanh rat nhieu nghien cuu trg giup, viec dieu tra khao sat he vi sinh vat tai noi nhiem la mot phan quan trgng Vai thai gian gan 40 nam de thich nghi, he vi sinh vat tai cac vijng bi nhiem chac chan da tao cho minh nhCrng ca chi thich ung dk giam thieu doc hai va phat trien Dia ban san bay Da Nang timg la nai tap ket va phan phat chat da cam/dioxin su dung de phun rai tai cac tinh mien Trung Viet Nam nen co nhiem doc cao gap vai tram den hang nghin Ian muc cho phep Dia diem da tra noi du tru da dang nguon vi sinh vat vai kha nang thich ung cao doi voi chat doc va hon the nua la cac loai co hoat tinh phan huy chat doc cung co ca hoi dugc nhan len Nhung tien bg ITnh vuc sinh hgc phan tu dung nghien cuu vi sinh vat cho phep nghien cuu tinh da dang va muc phan bo cua vi sinh vat moi truong song mot each hieu qua hon, dac biet la co the tiep can dugc phan Ion vi sinh vat hien chua dugc nuoi cay va phan lap phong thi nghiem (tren 90%) Vai thai gian phoi nhiem chat doc rat dai, tai khu vuc nhiem doc a san bay Da Nang mot quan the vi sinh vat (ca hieu va ky khi) co kha nang tieu doc chac chan da dugc tich luy sau thai gian dai, lam nguon nguyen lieu qui gia cho cac nghien cuu khoa hgc (theo trao doi voi TS Do Ngoc Lanh, Trung tam Nhiet dai Viet Nga - Bg Quoc phong) Du an ''Khao sat vi sinh vat tai vung bi dnh huang dioxin vd ddnh gid tiem ndng tieu doc cua vi sinh vat bdn dia" dugc de xuat nham tim hieu nhirng loai vi sinh vat co kha nang tham gia phan huy chat doc tai khu vuc nhiem (mot each true tiep hay gian tiep) cung nhu cac gen tham gia vao qua trinh Ket qua cua viec thirc hien nhiem vu neu tren se dugc su dung lam tai lieu khoa hgc tham khao cho ca quan nghien cuu tham gia viec xay dung cong nghe tiy doc dioxin Ben canh do, cac chung giong vi sinh vat dai dien phan lap dugc tu moi truong nhiem qua trinh thuc hien d\x an se dugc luu giO", bao quan Bao tang giong chuan vi sinh vat tai Vien Vi sinh vat va Cong nghe sinh hgc, lam ca so du lieu de nghien cuu va ung dung cho cac don vi quan tam ca sau khu vuc nhiem doc da dugc xu ly v lunn phi thyc hi^n dy an va nguon kinh phi 19 Gidi trinh cdc khoan chi (dan vi tinh: trieu ddng) Khoin Thue khoan chuyen mon (155 trieu ddng) TT Npi dung thue khoan Viet tong quan tai lieu, xiy dimg de cuong nghien cim chi tilt Kinh phi Thu mau v i xu ly mau tai dia dilm bi nhilm ddc Phan tich cic yeu td sinh dia hda va mdi trudng ciia miu thu dugc Phin tich cic yiu td thd nhuang cua mau thu dugc 5 Phan tich dp nhiem dpc cic mau dit Phin tich dp nhiem dpc cac mau trim tich thiiy vuc Phan tich VSV hiiu Prokaryote (vi khuin va xa khuin) cac mau thu thap tren mdi trudng dac hieu cho nhdm Phan tich VSV hiiu Eukaryote (nim men va nim sgi) cic mau thu thip tren mdi trudng dac hieu cho nhdm Phin tich VSV hiiu Prokaryote cic mau thu thip tren mdi trudng chura hgp chat tham da nhin lim ngudn cac bon va nang lugng nhit 10 Phin tich VSV hieu Eukaryote cic miu thu thip tren mdi trudng chua hgp chat tham da nhin lam ngudn cac bon vi nang lugng nhit 11 Phin tich vi sinh vit ky khu nitrat cic mau thu thip 12 Phin tich vi sinh vat ky khir sat III cic miu thu thip 13 Phin tich vi sirih vit ky khir sulfat cic mau thu thip 14 Phin tich vi sinh vit ky khir clo cic miu thu thip 15 Phin lip vi bao quan cic chimg VSV hieu dai dien va cic chiing cd nang phit trien tren mdi trudng chira chit ddc 16 Phin lip va bio quin cac chimg VSV ky dai dien 17 Giii trinh tu cac gen phin loai (rADN) ciia cac chimg VSV dai dien va cac chiing cd kha nang ung dung 18 Nghien cuu cac dac diem sinh ly, sinh hda ciia cic chiing VSV hieu dai dien vi cac chimg cd kha nang ung dung 19 Phit hien gen dioxygenaza cic chimg VSV hieu phin lip dugc, tich ddng gen vi dpc trinh tu 20 Nghien curu cic dac diem sinh ly, sinh hda cua cac chiing VSV ky dai dien vi cic chimg cd kha nang ung dung 21 Phit hien gen khir clo cac chiing VSV ky phin lip dugc, tich ddng gen va dpc trinh tu 22 Toi uu hda phuang phip va tich chiet ADN tdng sd true tiep tir cic miu thu thap 23 Khio sat, so sinh phin loai vi sinh vat nhin sa (Prokaryote) cac mau thu thip bang phuang phip sinh hpc phin tu (PCR/DGGE) 24 Khio sat, so sinh phin loai vi sinh vit nhin thuc (Eukaryote) cic mau thu thap bing phuang phip sinh hpc phin tir (PCR/DGGE) 25 Thilt lap ngan hang gen dioxigenaza tren ADN tdng sd tir cac miu hiiu bing PCR su dung cic mdi dac hieu 26 Dpc cic trinh tu ngin hang gen dioxygenaza da thilt lip 27 Thilt l$p ngan hing gen khir clo tren ADN tdng sd tir cac miu ky bing PCR sur dung cac mdi die hieu 28 29 Dpc cac trinh tu ngin hang gen khir clo da thilt lip Xu ly du lieu dpc trinh tu cac ngin hang gen bang tin sinh hpc 30 Bio cio khoa hpc tdng ket du in Thue khoin lao ddng rira dung cu thi nghiem 20 000 d/ngay cdng x 100 ngiy cdng/nam x nam Tong: MQt tram nam mvoi lam trieu dong 10 31 155 Khoan 2: Nguyen vat li^u, nang luong (226 trieu dong) (chi tiet xem phin phu luc) Noi dung TT Hda chat Hda chit diing phin tich vi sinh vit Hda chit dimg phin tich cic chi tieu mdi trudng - Hda chit dimg sinh hpc phin tir Vat lieu tieu hao ' Dung cu thuy tinh nudi ciy VSV - Dau cdn, dng nhua eppendorf dung SHPT Cac thiet bi liy miu, bao hd lao ddng Nang lirong - Dien - Gas Tong: Hai tram linh sau trieu dong * * r Kinh phi 165 25 20 120 55 25 25 4.8 226 Khoan 3: Chi khac (69 trieu ddng) TT Npi dung Cong tac phi, ve may bay di lay mau - Ve miy bay: ngudi x 000 000 Taxi di lai tai Ha ndi & Di nang - Luu tni: phdng (4 ngudi) x 300 000/ngiy x - Cdng tic phi vi bdi dudng ddc hai cho cin bd tai hien trudng ngudi X 100 000/ngiy x Quan ly cor sd Chi phi danh gia, kiem tra, nghiem thu - Chi phi xet duy|t de cuang thdng qua hdi ddng - Chi phi nghipm thu giiia ky - Chi phi nghipm thu chinh thuc Kinh phi 13 1,8 1,2 11.250 20 2,5 2,5 - Chi phi hgi thao Chi phi khdc - Thdng tin lien lac (Fax, dien thoai) - Photo, in in tai lieu, ddng quyen - Van phdng phim - Cic chi phi khic Tong: Sau muroi chin trieu dong 10 24,75 1.5 1,5 16,75 69 20 Phin bo kinh phi thyc hien dy an theo cac khoan chi (dan vi tinh: trieu ddng) Nguon kinh phi TT Tong so Tong kinh phi Trong Thue Nguyen khoin chuyen mon vit lieu chuyen nang dimg lugng Thiet bi, Xiy Chi khic dung sua chiia nhd 450 155 226 0 69 450 155 226 0 69 0 0 0 0 15,3% Trong dd: Kinh phi sir nghiep mdi trudng Cic ngudn vdn khac -T\rcd -Khac 100% Tyl? 34,5% 50,2% Hd noi, ngdy 28 thdng 06 ndm 2007 CHU NHIEM DU AN -Q^ Duang Van Hgp Ngay >3 thing f nam 2007 THUTRLTCJNGDONVI (ky ten, dong ddu)^ PHO GIAM -xjOC Ngay \o thing f nam 2007 PHE DUYET CUA DHQGHN TL.GIAM DOC DAI HOC QUOC GIA HA N Q I TL.G'AM£)6c raNG BAN KHOA H0C-CD.N5 NGHE TSKH Nguyin Dinh Dire ''TT DAI HOC QUOC GL\ HA NOI : S6: 3Cib /KHCN CONG HOA XA HOI CHU NGHIA VIET NAM Ddc lip - fu - Hanh phiic Hd Npi, ngdy -l-fthdng ndm 2007 QUY^T DINH CUA GIAM D C DAI HOC QU6c GIA HA NOI VI viae ph6 duydt nhi6m vu quan ly Nha nude ve bao va mdi trudng nam 2007 cua DHQGHN GIAM D C DAI HOC QU6c GIA HA NOI Can cii Nghi dinh sd 07/2001/ND-CP thing nam 2001 ciia Chlnh phii vl Dai hpc Qudc gia; Can cu: Quy^t dinh sd 14/2001/QD-TTg 12 thing nam 2001 ciia Thii tudng Chinh phii vl viae td chiic lai Dai hgc Qudc gia Ha Ndi; Can cu" vao (Juy ch^ T6 chifc va Hoat ddng ciia Dai hpc CJuoc gia dugc ban hanh theo CJuy^t dinh sd 16/2001/QD-Trg 12 thing 2'nam 2001 cda Thii tudng Oiirih {rfiu; Can ci^ bian ban hdi dong xet duyat dl cuong nhiam vu quan ly Nha nude ve bao va mdi trudng nam 2007; Theo de nghi ciia Ong Trudng Ban Khoa hgc - Cdng nghe, DHQGHN QUY^TDINH Dilu 1: Pha duyat nhiam vu quan 1^ Nha nude vl bao ve mdi trudng nam 2007 ciia DHQGHN, ma sd QMT.07.02 "Khao sat vi sinh v|t tai vung bi anh hudng dioxin va danh gia tiim nang tiau ddc cua vi sinh vit ban dia" Thdi gian thuc hian: 24 thing (8/2007 - 8/2009) Tong kinh phi: 450 trieu ddng, kinh phi nam 2007 la 200 tridu ddng, kinh phi nam 2008 la 250 trieu dong iSty tir nguon kinh phi su nghiep kinh ta' (nhiam vu bao ve mdi trudng) ciia DHQGHN Dilu 2: B nhiam TS Ducfng Van Hcrp, Vian Vi Sinh vit va Cdng nghe Sinh hoc lam chu nhiem nhidm vu Chu nhifm nhiam vu chiu trach nhiam tri^n khai cac noi dung ciia nhiem vu theo dung tia'n da duoc pha duyet Dieu 3: Ong Chanh Van phdng, Ong Trudng Ban Khoa hgc - Cdng nghe va chii nhiam nhiem vu cd trach nhiem thi hanh quy at dinh Al HOC 9U6GJGL\ HA NOI \MD6C J(k^^^ Nai nhin: -NhuDilu - Luu VP, KHCN GS.TSKH Vu Minh Giang DAI HOC QU6C GIA HA NOI CONG HOA XA HOI CHU NGHIA VIET NAM ca S6: S^f Ddc lap - fu - Hanh phiic ========== /HD-KHCN HOP DONG NCKH THlTC HlfiN NHlfeM VU QLNN v f BAO V£ M I TRI/CJNG - Can cii Quy^ dinh s6 3050/KHCN 14 thang nam 2007 ciia Giam ddc Dai hoc Qu6c gia Ha N6i v^ viec phe duyat nhiam vu quan ly Nha nude ve bao ve mdi tnr&ng nam 2007 cua DHQGHN va bd nhiem chu nhidm nhiem vu - Can cii v^o ban de cuong thuc hien ciia nhiem vu ma sd QMT.07.02 va sd kinh phi da dugrc phe duyet Chung tdi g6m: BSn giao (Ben A): Ban Khoa hoc - Cong nghe DHQGHN DT: 7548664, Fax: 7547724 Dai dien: TSKH Nguyin Dinh Dufc Chiic vu: Trirdng Ban Khoa hoc - Cong nghe Va TS Vu Dinh Giap Chanh Van phong DHQGHN Ben nhan (Ben B): Dcfn vj cdng tac: TS Duong Van Hgp Trung tam Cong nghe Sinh hoc - DHQGHN li Chu nhiem de tai ma sd Hai ben thoa thuin nhu sau: Dieu 1: Ben B chiu trach nhiem td chiic trien khai cac ndi dung nghien ciiu nam 2007 - 2009 cu the dudd day: • Thu thap mau nhiem dioxin tai san bay Da nSng va xir ly mau trudc dua ve phdng thi nghiem dd phan tich cac ydu to sinh-dia-hoa va vi sinh vat " Nghien ciiu da dang vi sinh vat tai dia didm nhiam th6ng qua cac phuang phap vi sinh vat truyen thdng - Nghien ciiu da dang vi sinh vat tai dia didm nhilm thdng qua phuang phap phan tfch d^u van ADN dua tren ADN tdng sd tach trigt true tiep tii mlu diit/nudrc thu thap, nhan doan gen ma hoa rARN va phan tich bang dien di bien tinh (DGGE) - Phan lap v^ tinh sach cdc chung VSV dai didn cho cac nhdm chinh co mat tai mdi trudng nhiem Nghien cihi cac dac didm phan loai va hoat tinh sinh hoc cua cac chung VSV phan lap dugc - Nghien ciiu da dang gen chiic nang dioxygenaza va gen tham gia vao qua trinh khir clo thdng qua phan tich ngan hang gen suf dung cac doan moi dac hieu - Phdt hien gen chiic nang cac chung VSV dai dien da phan lap tai dia didm nhiem, tich dong v^ doc tiinh tu Diiu 2: Ben B ndp cho ben A cac san phim khoa hoc theo ndi dung va tien thuc hien cua dd tai theo dot: dot (trudc 3()/6^008) va dat (trudfc 30/6/2009) Dot 1: - Phan tich cac ye'u td smh dia hoa tai dia didm lay mau nhidm dioxin - Phan tich dd nhiem ddc cua mau thu thap li' - Khao sat, so sanh vi sinh vat tai viing nhidm dioxin nang va vdng khdng hoac it d nhidm bang phuang phap vi sinh truyen thdng ^ - Khao sat, so sanh ph£n loai vi sinh vat tai viing nhilm dioxin nang va vung khdng hoac it nhilm bang phuang phap sinh hoc phan tu Dot 2: - Danh gia tilm nang phan huy cua VSV ban dia thdng qua phan tich da dang gen chiic nang (dioxygenaza va gen tham gia clo) mSu nhilm - Phan lap, nghien ciiu dac diem phan loai va dac didm di truyin ve gen chiic nang tham gia khir ddc dioxin cua cac chung VSV dai dien cho cac nhdm chie'm da sd khu vuc nhilm Bieu 3: Kinh phi cua d6 tai tur ngudn su nghiep mdi trudng: - Kinh phi nam 2007: 200 trieu ddng - Kinh phi nam 2008: 250 trieu ddng Kinh phi bao gdm nhutig khoan ddng gop nghla vu theo qui dinh hien hanh cua Nha nude Sd kinh phi chu tri de tai se nhan tai Phdng Tai vu, Van phong DHQGHN Bieu 4: Ben B cd trach nhiem chi tieu kinh phi dugc ca[p theo dung muc dich, tiling che' dd tai chinh hien hanh va quye't toan vdi Phong Tai vu, Van phong DHQGHN dgt theo nam tai chinh: dgt (trudc 30/3/2008) va dgt (tnldc ng^y 30/3/2009) Sau kinh phi dgt da quy^t toan xong, chu tri dl tai sS dugc nhan kinh phi dgt ffi/tf5; Hai ben cam kSt thuc hien diing cac dilu khoan da dugc ghi hgp ddng Trong qua trinh thuc hien hgp ddng, hai ben phai thdng bao cho nhutig v ^ dl sinh v^ cilng ban bac giai quydt Biiu6: Hgp ddng lam ban, mdi bdn giii mdt ban, ban luu van phdng DHQGHN, ban giri de'n Phdng Tai vu Ca quan DHQGHN Hd Ndi, ngdy hi thdng 6nam 2007 DAIDlfiNBENB DAI DIEN BEN A TRU&NG BAN KHOA HOC- CONG NGHfe -^n^y TS.DirONGVANHOP TSKH NGUYfiN DINH HXSC PHONG DHQGHN DAI HOC QU6C GIA HA N Q I Vi$n Vi sinh v^t va Cong nghe sinh hoc CQNG HOA XA HQI CHU NGHIA VIET NAM Doc lap - Tur - Hanh phuc PHIEU DE NGHI THAY DOI TRONG QUA TRINH THUC HIEN DE TAI KHCN CUA DHQGHN Ten de tai: Khao sat vi sinh vat tai vung bj anh hudng dioxin va danh gia tiem nang tieu doc ciia vi sinh vat ban dja Ma so de tai: QMT.07.02 Chu tri da t^i: TS Duong Van Hgp Co quan chu tri: Vien Vi sinh vat va Cong nghe sinh hoc, Dai hoc Quoc gia Ha npi Nh&ng thay doi ve tien do, thdi gian nghien cuu: Xin gia han thdi gian nghiem thu dk tai thang (tu thdi han 30/6/2009 sang thdi han 30/10/2009) chat chuan noi de phan tich ham lugng dioxin bang sac ky khoi phd can nhap ngoai hien chua the dugc cung cap, lam dnh hudng den mot so ket qua nghien cuu ciia de tai Ngdy^fthdng C ndm 2009 Co quan chu quan duyet TL GIAM DOC Ngdy'iZ thdng ndm 2009 Co quan chu tri Ngdy thdng ndm 2009 Chii nhiem de tai TRUONG DAI HOC QUOC GIA HA NOI TRUING BAN KHCN Duong Van Hgp 6STS:»/^!«^ ^ ^^^ HANOI C O N G HOA XA H O I CHU N G H I A V I £ T NAM Dpc lap 'Tudo ' Hanh phdc HOC BIEN B AN HOP HOI DONG KHOA HOC NGHIEM THU CAP CO Sd DE TAI CAP DHQGHN MA SO: QMT.07.02 Ten detdi: Khao sat vi sinh vat tai viing bi anh hudng dioxin va danh gia tilm nang tiau ddc ciia vi sinh vat ban dia C/id/li: TS Duong Van Hgp Quyet dinh thdnh lap Hpi dong Khoa hgc nghiam thu dl tai sd 151/QD-CNSH 23 thing 10 nam 2009 Ngdy hgp Hpi dong: 28/10/2009 Danh sach cdc thdnh vien cua Hpi dong: GS TS La Dinh Luong Chii tich Hdi ddng TS Nghiam Nggc Minh Phan bian TS Nguyin Quynh Uyin Phan bien TS Dao Thi Luong Thu ky TS Nguyin Huynh Minh Quyan Uy vien TS La Thu Hiln Uy vian TS Nguyin Thi Hoai Ha Uy vien a Cd mat: 1- ngudi b Ving mat: Q ngudi Ngudi trinh bdy bdo cdo: Ho va tan: TS Dinh Thuy Hang Ket qud ddnh gid: Sd philu phat ra: ^ Sdphiluhgpia: > Kit qua: ^ fo~f '1 u •'"1 -CA_ IcZh 76 f Thao ludn cua Hpi ddng CKx? -f^/ dl hu phA l>!eu' '' - rx^ ^cn/, ] ^ M ^ y^-;;/, L^ ^^^ k^^u^ct^ui, ^ ^^lA£ 4f ^ CUA