Dd Thi Tuyen vd Dtg Tap chi KHOA HOC & CONG NGHE 86(10) 153 155 H0 4T TINH KHANG SINH CUA XA K H U A N TRONG D A T TAI CAc KHU VUC CO HOAT D O N G KHAI T H A C K H O A N G S A N flo Thi Tuyen, Luong T[.]
Dd Thi Tuyen vd Dtg 86(10): 153 - 155 Tap chi KHOA HOC & CONG NGHE H0.4T TINH KHANG SINH CUA XA K H U A N TRONG D A T TAI CAc KHU VUC CO HOAT D O N G KHAI T H A C K H O A N G SAN flo Thi Tuyen, Luong Thj Huong Giang, Dao Thj Hang Nguyen Thi Huong Lien, Vi Thi Doan Chinh" Tnrdng Dai hoc Khoa hoc DH I'lu'ii \guydi TOM T A T Til" 40 mau dat thu d cac dja diem khac tai cac khu vuc dang chiu anh hudng ciia hoat dgng khai thac khoang san ciia tinh Thai Nguyen, da phan lap dugc 162 chiing xa khuan thugc chi Streptomyces Qua kiem tra hoat tinh khang sinh (HTKS) ciia cac chiing xa khuan phan lap dugc c6 82 chiing (chiem 50,62%) cd hoat tinh khang cac vi sinh vat (VSV) kiem djnh Trong sd dd sd chung cd boat tinh khang vi khuan Gram (+) la cao nhat - cd 61 chung (chiem 74.39%) cd 58 chiing (chiem 70,73%) cd boat tinh khang nam va thap nhat la sd chung cd hoat tinh khang vi khuan Gram (-) chi cd 37 chimg (chiem 45.12%) Dac biet cd 31 chiing (chiem 37.8%) cd hoat tinh vdi ca nhdm vi khuan Gram (+) va Gram (-), cd 20 chung (chiem 24,39%) cd hoat tinh vdi ca nhom vi khuan Gram (+) vi khuan Gram (-) va nam mdc Vdi muc dich tuyen chgn dugc cac chung co hoat tinh khang nam, chimg tdi da lira chgn chiing cd boat tinh manh nhat de tiep tuc nghien cuu la TC13.1, TC13.2, TC12.1, HT17.8 va HT17.9 Til' khoa: chdt khdng sinh chung, khudn tv hoat tinh khdng sinh xa khudn DAT VAN DE Chat khcing sinh (CKS) dii Vci dang dugc sir dung rdng riii nhieu linh vuc khcic NgOcii muc dich y bgc CKS cdn dutic sir dung chiin nudi tlui y va dac biet la cdng tac bao ve tlurc Vcit de giam dcin viec sir dung cae loai thudc hda hoc Song, viec sir dung CKS cimg rgng rai bao nhieu thi nguy CO lam xuat hien cua cac vi sinh viit khang thudc lai cang tang len bciy nhieu Dieu khien cho Ccic CKS thudng diing khdng cdn tae dung nua Chinh vi vay nhimg nghien ciru de tim kiem phcit hien cac CKS mdi lit tir nhien ludn la nuit v'eu Cciu eiip thiet Vci cd y nghla thiet thuc Trong sd cac ddi tugng de tim kiem CKS thi XCT khuiin ki ddi tuting ludn dutic ciui y nhieu nhcit vi cd tdi 70% sd ehung xa khuan cd kha niing sinh CKS Fliiii Nguyen ki nuit tinh nit giciii tiem niing ve ndng, lam nghiep nen cd he VSV khci phong phii Ddng thdi Thai Nguven cting niim viing sinh khoang cd nhieu loai hinh khoang san phan bd tiip trung Cac hoat ddng khai thae khoang san da ed nhiing tae ddng dang ke den mdi trudng dat, nutie vii qua dd, rat cd the se cinh hudng den he VSV diit ti nhirng khu vuc ma hien van chua duge nghien ciru Trong bcii bao luiy, cluing tdi thdng bao nuit sd ket qua bude dau khao sat va danh gia boat tinh sinli hoc cua nhdm xa khuiin d diit tai eac khu vuc dang chju anh hudng ciia boat ddng khai thiic khoang san tren dja biin tmh Thiii Nguyen NGUYEN LIEU NGHIEN CUU vA PHUONG PHAP Nguyen lieu - 40 mau dcit kiv tir cac dja diem khiic d khu vu'c niii Phao, Hii "Fhirgng, Dai Tir va Md Scit Trai Cau, Ddng Hy thude tinh Fhcii Nguyen Diiv lii nhung khu vuc dii vii dang cd cac hocit ddng khai thac khocing san nhieu iicim - cluing VSV kic'in djnh: Escherichici coli V'FCC-B-883 rseiidomoncis aeruginosa ' I'd: 11987 123606 Email: Yichinli5'agiiiail.com V'FCC-B-481, Bacillus subtilis VFCC-B-888 153 86(10): 153- 158 Tap chi KHOA HOC & CONG NGHE Dd Thi Tuyen vd Dtg KET QUA v A THAO Etisarium oxysporum VTCCF-1301 A.spergillus niger VTCCF-001 Fusarium solani VTCCF-1302 Vien Bao tang giong chuin vi sinh vat cung cap; Staphylococcus aureus ATCC 25923 Vien Kidm Nghiem Bd Y te cung cap LUAN Hoat tinh khiing sinh cua xa khuan Til' 40 mau dat lay tii' cac dja diem khac tai cac khu vuc dang chju anh hudng ciia hoat dgng khai khoang d tinh Thai Nguyen, chimg toi da phan lap va thuan khiet dugc 162 chimg xa khuan thugc chi Streptomyces So lugng va su phan bo ciia xa khuan dugc trinh bay tren bang - Cac mdi trudng: Cause de phan lap va nudi cay xa khuan; mdi trud'ng MPA va PDA de nudi cac chung VSV kiem djnh Ket qua bang cho thay sd lugng xa khuan cac mau dat la kha phong phu va phu thugc nhieu vao dac diem, tinh chat ciia tirng loai dat Sd lugng xa khuan gap nhieu nhcit d cac loai dat mau (12,07 x 10^'CFU/g) va dat vudn (10 x 10'''CFU/g) Ket qua hoan toan phii hgp vdi dac diem sinh hge eua xa khuan va dac diem canh tac dat tai nhung khu vuc lay mau Xa khuan thudng phan bd nhieu d cac loai dat to'i xdp thoang khi, giau chat hiiu CO', cd pH trung tinh va am thich hgp Phu'ong phap nghien ciiu - Thu mdu ddt, xdc dinh pH cua ddt vd phdn lap xg khudn [2], - Xdc djnh mdu sdc cua he khudn ty [5] - Xdc djnh hogt linh khdng sinh (HTKS) theo phuong phap thdi thach de so' tuyen xa khuan va phuong phap due Id de sang lgc xa khuan - Xg khudn dirge len men tren mav' lac trdn 220 vdng/ phiit, thdi gian len men 120 gid d 28"C Bang L Sd lugng vii su phan bd xa khuan dat Loai dat lav mau pH So lu'ong mau Dat chti ciia dat So luong XK/g (CFU/g) So chiing XK phan lap 4.12 1.7 X 10' 11 Dat keo 5,53 5.3 X 10" 26 Dat mau 7,22 12,07 X 10' 44 Dat liia 6,70 4.2 X 10' 11 Dal \ iron 7,30 lOxlC* 55 Dat doi tree 4.15 3.6 X 10" 15 Tong cong 162 40 Bang Sd lugng va ty le cac chiing xa khuan cd HTKS phan theo nhdm mau TT XK phan theo nhom mau XK phan lap du'o'c So iu'ong Ty le (%) XKco HTKS Soluong Tyle XKco HTKS Ty le (%) so vo'i tong so chiing phan lap du-oc (%) Xam 83 51.23 38 46.34 23.46 Trang 39 24.07 22 26.83 13.58 Xanh 13 8.02 4.88 2.47 Nau 12 7.41 7,32 3.70 Hong 5.56 8.54 4.32 Tim 2.47 3.66 1.85 Vang 1.23 2.44 1.23 Tong 162 100 82 100 50,62 154 Dd Thi Tuyen vd Dtg Tap chi KHOA HOC & CONG NGHE Dat trdng mau Vci dat vudn thudng xuyen dugc cude xdi, bd sung ngudn phan bdn huu CO va vd CO, cd pH trung tinh, dd la cac dieu kien rat thuan lgi cho sir phat trien eiia xa khuan Ngugc lai, nhung loai dat ngheo dinh dudng va dac biet lii ed pH qua cao hay qua thap, xa khuan phan bd rat it Theo ket qua nghien ciru, dat chju anh hudng cua cac hoat ddng khai thac thiec d khu vuc mii Phao hay khai thac quang sat d Trai Cau deu thudc loai dat chua [3], vi vay khdng thich hgp cho xa khuan phat trien Tuy nhien nhiing noi ed hoat ddng san xuat eua ngudi, pH cd the dugc cai thien, Mau sac ciia he khuan ty Xci khuan cung riit da dang Chimg tdi nhan dugc nhdm miiu Sd lugng va ty le cae nhdm mau rat khac dugc the hien tren bang 86(10): 153 - 158 Ty le cac chung xa khuan cd HTKS tuo'ng ddi cao, cd 82 tdng sd 162 cluing dugc kiem tra, chiem 50,62%) So sanh vdi eac ket qua da cdng bd [1.2.3.4] day la mdt tv le khci eao Dieu dii chirng td sd lugng xa khuiin cd kha nang sinh CKS d cac khu vuc cd boat ddng khai thac khoang san khdng nhirng khdng thap ma cd phan eon cao han so vdi nhieu khu vuc khae d Thai Nguyen [1.2], Ket qua tren bang va hinh I cdn cho thay ty le xa khuan cd HTKS eiing rat khac giira cac nhdm mau Nhdm mau xam ed sd lugng chiing nhieu nhat ddng thai cung cd ty le chimg cd HTKS cao nhat (chiem 46,34%) Tuy nhien neu xet rieng tirng nhdm mau cac nhdm mau vang, tim va hdng, mac dii sd lugng chimg it nhung ty le chung cd HTKS lai rat eao Day la ket qua dang luu y Pho khang sinh ciia cac chiing xa khuan phan lap :"^-i'>'i 4,''0' ^^^k \ an-' X.Mlll Nhom mau Hinh I Ty le xa khuan cd HTKS phan theo nhdm niciu Nhu thudng le xa khuan tluicie nhdm xam va trang viin chiem uu the Ty le xa khuan thudc nhdm Xcim chiem 51,23%, tiep theo ki nhdm trang chiem 24.07%, cac nhdm niciu cdn lai chiem ty le riit tluip Kiem tra kha niing ire che Ccic nhdm VSV kiem djnh, ket qua the hien tren bang va hinh da cho thay tinh ddi khcing cua Ccic chimg xa khuan vdi Ccie VSV kiem djnh rcit khac Trong tdng sd 82 chimg ed HTKS, nhu thudng le, sd chirng cd kha nang khang vi khuiin Gram (-I-) la cao nhat, cd 61 chimg chiem 74,39%, tiep theo la khang nam mdc, ed 58 chiing - chiem 70,73% va thap nhat la khcing vi khuan Gram (-), chi cd 37 cluing chiem 45,12%, Dcoccio: aureus A tCC 25923 54 65.85 liacilUis suhn'li.s V rCC-R-888 45 54,88 F.sc icnchia coll VTCC-B-883 32 39,02 P.sc idomonas aeruginosa 22 26.83 Fu.s iriuni oxysporum V rCCF-1301 42 51,22 Lu., iniim solani VTCCi •1302 22 26.83 •Ispergillus niger V TCCl -001 32 39.02 V"rCC-H-481 B a n g s HTKS cua 10 chung xa khuan tuyen chgn lloal tinh khang sinh (I)-d, mmm) Kv hieu chung /;" V i c e -B-88: /-*, aeruginosa vrcc-u-481 ll siihlilis NKC-W 888 TC 13.1 24.5 I 0.5 28.3 1- 0.4 23.5 ^ 0.5 T( 13.2 26.4 :x 0.5 2>).S • 0.4 24.2 :r 0."- TC 15.9 27.2 ±0.3 27.3 10.4 T( 12.1 26.3 -: 0.6 TC 16.11 10.8 ± " /•, imsporiiiii vrcci-1301 /•", solani l,/»gc7' vrccF- vrc'ci"- L302 001 22.6 ± 0.5 26.4 ± 0.5 25.9 ± 0.3 - 24.5 - 0.5 24.8 ±0.3 26.0 1- 0.5 - 15.7 3:0.5 II.2 0.3 8.9 : 0.4 12.1 ± 0.4 - 20.7 ,- 0.3 20.7 ± 0.3 27.1 ^0.4 25.1 rO.4 24.6 r 0.5 - 8.9 i 0.4 l O X • i).y 13.1 ^ 0.4 15.4 -0.5 16.5 : 0.5 - V, (iiireiis AICC 25923 III 17.8 10.2 : 0.1 1.2 • 0.1 10.2 r 0.2 10.2 + 0.1 15.1 ±0.1 16.8 0.7 - HI !') 20.8 - 0.7 20.5 0.5 17.2:0.3 15.2 0.2 23.8+ 0.7 27.8 ±0.7 - i n 12.2 12.2 15.3 ! 0.5 11.2 ±0.1 15.4 ±0.3 17.5 ±0.5 1.2 ±0.1 - 111 19,2 10.3 ± 0.3 10.2 ±0.2 - 13.2 ±0.1 5.0 ± 0.4 15.2 ± 0.2 - III r , i s 15.3 r 0.3 6.0 t 0.7 15.3 I 0.3 - - 9.2 ±0.1 - Li6 T 0.2 Dd Thi Tuyen vd Dtg Tap chi KHOA HOC & CONG NGHE 86(10): 153- 158 Hinh Kha nang ire che VSV kiem djnh cua mdt so chung \a khuan 1:TCI6.12: 2:TC15.9: 3:TCI3.1: 4:TCI3.2: 5:TCI2.1: 6: I IT 19.1: -':HTI7.H: 8: LIT T IS Tuyen chon cac chiing xa khuan co boat tinh khang sinh eao Dong Hy Tluii Nguven dcio ddng khoang til 1.7 X 10'' den 12.07 x IO'' CFU/g dcit Ccin cir tir eae kc't qua kiem tra so' bti ve HTKS d tren, chung tdi chgn 10 chimg cd boat tinh cao, hocit phd rdng de tiep tuc Scing Igc Cac chimg dutic nudi lae mdi truing (iause I djeh the \ii kic'in tra IFFKS cua djeh len men bang phuong pluip due ki Ket qua dugc the hien d being Ket qua tren cho thii> phcin Idn ciic cluing lira chgn deu Vcan giii dugc liocat tinh chuyen tir mdi trudng thach sang mdi trudng djeh the Hoat tinh ciia Ccic ehiing tuong ddi dn djnh Tuv nhien tit ca cae ehung deu khdng cd liOcit tinh vdi luim 'ispergillus niger VTCCF-001 Cd ehung ed boat tinh manh nhcit cd kha nimg ire ehe duge vdi cluing VSV !;:^m djnh: TCI3.1 FC13.2^' TCI2.1 HT 17.8, iii ;9.1 (hinh 4) Dii pluin kip \ci thuan khiet dirge 162 ehung XcT khuan tir 40 nuiu diit tlui tai nuit sd khu vuc dill bi nhiem qtuing eua tinh Fhiii Nguven dd cd 82 ehung cd boat tinh khiing sinh (chiem 50,62%), 61 ehung (74,39%) khang vi khiicin Gram ( + ), 58 chung (70,73%) khimg luim mdc, 37 chimg (45,12%) khcing vi khiiim Cjram (-), 31 chimg (37.8%) ca nhdm vi khuan kiem djnh G+ Vci G- 20 chung (24.39%) khcing ea vi khuan (+), vi kluiim Gram (-) vii niim mdc Vdi lurting i-,,ul-,ien ciru tuyen chgn eac eao t d bocit phd rgng Vci dcic bu" ^•>^ imli khcing niim giiv' benh thuc \at chimg tren dugc lira ebon de tiep tuc nghii':i ^iiu KF.TinAN I So lugng xa kiiiian phim bd a diit lai khu virc mii Phiio Dai Fir vii mo sat "I'rai Cau |1], Vi Thj Doan Chinh (2011), Tuyen chon vd nghien ciru xg khudn cd kha ndng doi khdng vdi mdi sd cluing vi khudn gd}- nhiem Iriing benh vien Bao Ciio tdng ket dc tcii khoa hge \ ii cdng nghe cap Bg Mci sti: B2009-TN07-02 |2] Vi "rhj Doan Chinh, Trjnh Nggc Hoiing irinh Dinh Kha, Vii rhi Lan (2007) Nghien ciru sg pluin Iv') cua xg khudn sinh clidl khdng sinh phdn lap tir (ldl Thdi Nguyen Bao ciio khoa hge ldi Da tiiven chgn ehung xa khuiin TC 12.1 T c ' l l TC 13,2 IFF 19.1, HT17.8 cd boat tinh khimg Ucim nuanh {F oxysporuni VTCCF-lSOha F solani VTCCF-13()2) su dung cho cac nghien ciru tiep theo TAI LIEU THAM KHAO 157 Dd Thi Tuyen vd Dtg Tap chi KHOA HOC & CONG NGHE nghi toan qudc NCCB khoa hoc su sdng Tr.433-437 [3], Le Dire Nguyen Qudc Viet (2007) Tdc ddng cua hogt dgng khai thdc khodng san tgi Dgi Tir Ddng Hy, Thdi Nguyen den mdi trudng khu vice Hdi thao KH Qudc gia "Nhirng van de mdi trudng va phat trien ben vung viing ddng bac Tr 153-159 [4] Nguyen Lan Dung, Pham Thj Tran Chau, Nguyen Thanh Hien, Le dinh Luo'ng, Doiin Xuan Mirgu Pham Van Ty (1978) Mdt sd phirong phdp 86(10): 153- 158 nghi&n cihi vi sinh vgt hoc Tap III, Nxb KH&CN, HaNgi [5] Biii Thj Viet Ha 2006 Nghien cim xa khudn thude chi Streptomyces sinh chdt khdng sinh chdng ndm gdy benh thgc vdt (x Viet Nam, Luan an tien sT sinh hge Ha Ndi [6] Nguyen Huynh Minh Quyen (2011) Dieu tra nghien ciru mgt sd boat chat cd kha nang khang vi sinh vat va khang ddng te bao ung thu tir te bao xa khuan Bao cao tdng ket de tai KH&CN cSp DH Qudc gia Ma sd 09.48 Ha Ndi SUMMARY A N T I B I O T I C A C T I V I T Y O F S O M E A C T I N O M Y C E T E S I S O L A T E D F R O M SOIL WITH MINING OPERATIONS Do Thi Tuyen, Dao Thi Hang, Luong Thi Huong Giang, Nguyen Thi Huong Lien, Vi Thi Doan Chinh' College of Sciences - TNU \ total of 162 actinomycete strains of the genus Streptomyces were isolated and pured from 40 soil samples collected from Thai Nguyen province These isolates were then examined antibiotic activities by agar streak method against Gram-positive (Bacillus subtilis Staphylococcus aureus) Gram-negative (Escherichia coli, Pseudomonas aeruginosa ) bacteria and fungi (Fusarium oxysporuni, Aspergillus niger Fusarium solani) Results indiciited that 82 isolates exhibited antibiotic activity (50,62%) Among them 61 strains (74.39%) possessed antibiotic activity against Gram positivie bcacteria 37 strains (45,12%) - against Gram negjitive bacteria, 58 strains (70,73%) - against fungi In particular, there are 31 strains (37.8%) against both Gram (±) and Gram (-) bacteria 20 strains (representing 24.39%) against all three groups of Grcim (+) Gram (-) bacteria and fungi Five strains signed cas TC 12.1, TC 13.1, TC 13.2 HT 19.1, HT 17.8 and HT 12.2 with high active against fungi were selected for further investigation Key words: antibiotic, strains, mycelium, antibiotic activitv actinomvcetes ' Tel: 098^ 123606 Email: vichinlo'crgmaiLconi 158 ... xg khudn [2], - Xdc djnh mdu sdc cua he khudn ty [5] - Xdc djnh hogt linh khdng sinh (HTKS) theo phuong phap thdi thach de so'' tuyen xa khuan va phuong phap due Id de sang lgc xa khuan - Xg khudn... Dieu dii chirng td sd lugng xa khuiin cd kha nang sinh CKS d cac khu vuc cd boat ddng khai thac khoang san khdng nhirng khdng thap ma cd phan eon cao han so vdi nhieu khu vuc khae d Thai Nguyen [1.2],... khang dugc ca nhdm vi khuan Gram (+) Gram (-) va nam mdc Bang Tinh ddi kh ang ciia xa khuan vd cac nhdm VSV kiem djnh Xci khuan cd HT voi VK Gr(+) Xci khuan cd HT vdi VK Gr(-) Xa khuan cd HT vdi niim