Top chi Kboa hgc Tnrang Dai bgc Cdn Tha Phdn i Khoa hgc Tunhien Congnghe va Moi Inr&ng 28(2013) 86 92 Tap chl Khoa hoc Tru''''dng Dai hoc Can Thd website sj ctu edu vn PHAN LAP VA NHAN DIEN VI KHUAN SAN[.]
Top chi Kboa hgc Tnrang Dai bgc Cdn Tha Phdn.i: Khoa hgc Tunhien Congnghe va Moi Inr&ng 28(2013) Tap chl Khoa hoc Tru'dng Dai hoc Can Thd website: sj.ctu.edu.vn PHAN LAP VA NHAN DIEN VI KHUAN SAN XUAT CHAT KET TU SINH HOC TRONG N i r c RI RAC VA iTNG DUNG TRONG XU" LY NlTOfC Ri RAC Cao Nggc Diep' va Pham ST Phuc' Viin Nghien cuu vd Phdt triin Cdng nghi Sinh hgc, TrudngDai hoc Cdn Tha ' Trudng Phd thdng Trung hoc Thanh An, huyin VTnh Thgnh, Thdnh phd Cdn Tho Thong tin chung: Ngaynhdn: 05/05/2013 Ngdy chdp nhgn: 29/10/2013 Title: Isolation and Identification biqflocculant - producing bacteria in landfill leachate and their application for leachate treatment Td khda: Enterobacter sp., Klebsiella sp., kit lu sinh hoc, nudc ri rdc, ti li kit tu Keywords: Bioflocculation, Enterobacter sp., flocculating rate, Klebsiella sp., sewage water ABSTRACT Bioflocculanfs produced by microorganisms canflocculateand settle dispersed organic particles so thai they can be usedfor wastewater treatment and it is not toxic to human and the environment Fifty-five bioflocculant-producing bacterial isolates were isolatedfrom ten leacheate samples offivelandfills from Vinh Long, Can Tho and Hau Giang Two isolates (Pl.l and P2.4) in 55 isolates had the highest flocculani rate and they were chosen to sequence randomly by automatic sequencer DNA sequencing were compared with Gen Bank database ofNCBl by BLAST N software The results showed that Pl.l isolate was 99% of the identify with Klebsiella sp NBRC 100048 and FJ57797 Klebsiella sp T'6-1: P2.4 Isolate M-as 98% of the identify with Enterobacter sp.A2 andJN6957I9 Enterobacter sp TX2 (2011) The optimal medium consist of starch YE CaCh for Pl.l isolate: sucrose, NH,Cl K2HPO.i+KH2POjfor P2.4 isolate with kaolin solution after minutes at pH=4 (Pl.l isolate) and pH=5 (P2 isolate) together with CaCh, 0.1% inoculanl (bacterial liquid) Applying these two isolates for landfill leachate freatment showed that iheyreduced the total solid suspension (TSS) and Chemical Oxygen Demand (COD) by 12.09% and 19 84% (Pl.l isolate): 12.4% and21.89% (P2.4 isolate) respectively T M TAT Chdt kit tu sinh hgc dugc san xudt tic vi sinh vdt, chung kit bdng vd lang tu cdc chdt huu ca di xir ly nudc thdi vd khong ddc hgi cho ngudi vd mdi tnt&ng Ndm muai ldm ddng vi khudn cd kha ndng tgo chdt kit tif sinh hgc dd dugc phdn lap lit 10 mdu nudc ri rdc thu thgp d bdi rdc cdc tinh Vinh Long, C^dn Tha vd Hdu Giang Trong dd cd hai ddng vi khudn PI I vd P2.4 tgo chdt kit lu sinh hgc protein cd li le kit lu cao vd chung dugc chgn di gidi trinh tu vd su dung phdn mim BLAST N so sdnh trinh tu cdc ddng vi khudn ndy vdi trinh tif cdc ddng vi khudn cd ngdn hdng du liiu NCBI, kit qua cho thdy ddng Pl.l cd ti li ddng hinh vdi dong Klebsiella sp NBRC 100048 vd FJ577970 Klebsiella sp T-6-} Id 99% dong P2.4 co ti le ddng hinh vdi ddng Enterobacter sp A2 vd JN695719 Enterobacter sp TX2 (2011) la 98% Moi trudng tdi mi cho ddng PI Id tinh bdt, yeast extract, CaCh cho ti li kit tu din 88.58%: ddng P2.4 Id sucrose, NH4CI, K2HP04+KH2PO.I cho fi H kit tu 86.81% vdi dung dich kaolin sau 5phiit di ldng dpH=4 (ddng Pl.l) va pH=5 (ddng P2.4) bd sung CaCh vd 0,1% liiu luong dung dich chira vi khudn Ifng dyng hai ddng vi khudn ndy xii ly nudc ri rdc ldm gidm luang TSS vd COD Id 12,09% vd 19.84% (ddng PI 1); 12,4% vd 21.89% (ddng P2.4) Tap chi Khoa hoc Tru&ng Dgi hgc Cdn Tha PhdnA: Khoahgc Tunhien Cdng ngfte vd Mat truang- 28 (2013/- 86-92 GIOI THIEU Kit tu sinh hgc Id sdn phim ciia hgp chdt cao phin tir dugc hmfa thdnh frong qud trinh ph^ trien cua vi sinh vit (Shih et al., 2001) Vi khuin san xuit chit kit ty sinh hgc Id nhiing lodi \ i khudn cd thi sir dyng cdc cfait dinh dudng frong mdi trudng dl tong hgp cdc fagp cfadt da phin tir ttong tl bdo dudi sy hogt ddng ciia cde enzjme die biet, sau dd chiing ed tfae dugc bdi tilt ngodi vd Idn tgi frong mdi tmdng faoac frln bi mdt vd te bao vi khudn, cic hgp chit ndy cd khd ndng tao sy kit ty vdi cdc cfait khde vd tgo mot kfadi nfady Idng xudng ddy Chdt kit tu dugc phin loai thinh ba nhdm cfainh: chit kit ty vfi co nfau alumuium sulfate \a polyaluminum chloride, chat kit tu tdng hgp hihi CO nhu la dan xuit ciia polyacr>lamide vi polyethyleneimine chit kit tu \a\ frong ty nfailn nfau: chitosan, sodium alginate vd ket tu sinfa fapc bdi vi khudn (Zhi-qiang et al 2007) Cac chit kit ty vo CO vi chdt kit ty tdng hgp hihi co cd khd ning gi\' ung thu, tic ddng mgnfa den tfain kinfa, cd thi giy d nhiem tfaii cdp frong qud trinh dp dung thyc tl vi chiing khd bj phin hu>' ttong tu nhien (Yokoi el al., 1994) Gin da\ kit ty sinh hpc bdi vi sinh vat dugc ngdnh cdng ngfal sinh hpc mdi tmdng chii y bdi vi chiing cd khd ndng pfaan gidi sinh hgc vd khfing dpe hai (Lu et ai, 2005) Theo Sheng et al (2006) cfait kit tu sinfafagccd tic dgng nhanh chdng vi an toan khdng ddc hai cho ngudi va mdi trudng Chdt kit ty sinh hpc cd nhieu loai nhung dupe quan ldm ngfailn ciiu nfaiiu ti polysaccharide, protein \a glycoprotein Trong xir ly nudc thai, chdt kit tu sinh fagc lir cde ngudn vi sinh vdt dugc sir dyng de xu ly dung djch thufic nfaudm (Zang et al 2002: Deng et al., 2005) cdc cfadt lo tirng vd ea (bentonite, dit set, Ca(OHh, aluminum oxide) (Levy et al., 1992; Shih et al., 2001; Yim et al 2007), acid humic (Zouboulis et al 2004) va cac chdt lo lung khdc (Salehizadeh et al 2000) Sy kit tu cdc vit chit niy giiip tam gidm cdc cfai lilu nfau COD vi mdt phdn dp due cua nudc thdi tmdc nuac thdi dugc xii ty bdng pfauong pfadp kfadc (Gong et al., 2008) Trong ngfailn ciiu ni}' chiing tdi trinh bay kit qud pfain lap, hieu qud kit tu ciia cdc dong vi kfaudn vira phdn lap, tdi im hda cac jeu to infa hudng den hieu qud kit ty suih hgc ciiafaaidong vi khuin cd ti le kit ty cao (> 80%) va img dung chiing xu ly nudc ri rdc die biet tara gidm ham tugng COD va TSS frong thinh pfaan nudc ri rae PHU'ONG PHAP NGHIEN c u r 2.1 Vdt lieu - Nuo€ ri ric tfau tir 05 bai rdc ciia Hoi Phii, Trd On (tinfa VTnfa Long) Thot Ndt, Cd Dd (Thanfa pfad Cin Tho), Tin Long (tinfa Hdu Giang) Mfii trudng pfain tap vi khuin sin xuat chdt kit ty sinh hpc polysaceliaride (Deng et al., 2002) vi mdi truomg phin lap vi kfaudn sdn xuit chdt kit tu sinh hpc protein (Hazana et al 2008) 2.2 Phinmg phap 2.2.1 Phdn lap vi khudn sdn xudt chdt kit tu sinh hgc Mudi mdu nudc ri rdc dugc pha lodng lir 10- lO"*; mdi pha lodng hiit 20 pi trdi diu trin moi trudng thach, sau dd li — d 30'*C Ticfa rdng timg kfauin lgc bang each cay chuyen tilp sang moi tmdng thach vdi lan, quan sit thiy kfaudn lac rdi diu tilnfainfacay cfauyin sang dng Kl tiep kilm fra rdng frln kinh failn vi xem nfau mgt ddng (isolate) rieng biet 2.2.2 Kiem tra hiiu qud ket tu sinh hgc cua cdc ddng vi khudn phdn lap Mdi dong vi khudn dugc nufii frong 50 ml mfii trudng ldng binh tam gidc 100 ml (mfii tmdng dimg dl pfain lap kfadng bd sung agar) Lie frln miy lie xoay vong a.tdc dp 150 vdng/pfaiit Nudi frong ngdy d nhilt dp pfadng cho vi khudn phit trien sdn sinh cfadt kit ty sinfa hpc Kiem tra khd ndng kit ly ciia cic ddng vi khudn bdng hdn hgp gfim 90 ml dung dich Kaolin (5 g/1), 10 ml dung dich CaCh 1%, pH =7, 100 pi dich vi khudn nudi cd mat sd >10^ te bdo/ml (ti 1^ 0,1%) Hdn hgp dugc khuay diu de ldng phiit sau liy phin ttong d tten dem OD d budc sdng 550 nm Miu ddi chimg diyc hiln tuang ty nhung kfadng chiing vi khudn Ty le kit ty dugc tinh theo cdng thiic: ODdo, -ODm, - X 100% ODd 2.2.3 Nhdn diin ddng vikhudntgo kit tu sinh hgc cho hiiu qud cao bdng phuang phdp sinh hgc phdn tir Nhtmg ddng vi khudn cd ti 11 kit tu sinh hgc cao dugc chgn dl nhdn dien bang each thyc hien cac phdn iing PCR de xdc dinfa gen 16S rRNA vdi cap mfii 27F va 1492R {Kim et ai, 2010): sdn phdm PCR se dien di tten gel agarose 0.8% vd phd Tap chi Khoa hgc Tru&ng Dai hgc Cdn Tho Phdn A: Khoa hgc Tu nhien Cong nglii \-d Moi tru&ng 28/2013/ diln di sdn p h ^ PCR dugc cfaypfainhqua GelDoc vdi thang cfaudn 100 bp sau chiing dugc gidi ttinh ty d cdng ty MACROGEN (Hdn Qudc), kit qua ttinh ty dugc so sdnh voi ddng vi khudn chuin d ngin hang diJ lilu cua NCB! bing phin mim BLAST N dl nfagn diln ddng vi kfauin dugc giai trinh ty gen 16S rRNA Xdy dyng cay pha he ciia cfaung bing phdn mim MEGA 5.05 (Tamura et ai, 2011) voi phuong phdp neighbor-joining dya tren 1000 Idn lap lai (bootsttaps) 86-92 cfaofaieuqua kit tu cao nfadt Ty le kit tu dugc linh gifing nhu md td d ttin Dp pH ciia dung dich Kaolin dugc chinfa frong khodng tir 3-9 bang HCl va NaOH Cdc mudi KCl, NaCl, CaCt^ MgSq4, MnS04, FCCIB, Al2(S04)3 dugc sir dung nhu ngudn cation vdi ndng dp va lieu lugng nfau CaCb Lilu lugng dich vi khudn nufii dugc bd sung dieu chinh tuong ling vdi cac ti 11 0,01, 0,02, 0.04, 0,06, 0,08 vd 0,1(%) Chudn bi mdi frudng nufii cay vi khuin vdi sy phdi hop ba ngudn carbon, nitrogen va muoi khoang theo diing ti 1? cac phan frong Bing 1, ta dugc 48 nghiim thiic khac ve mfii tmong nufii vi khudn Nufii vi khuan ttong dng nghiem loai 10 ml d nhiet dp phdng, lie 160 vdng/pfaiit Sau ngay, kiim tra hi?u qud kit ly sinh hpc vdi Kaolin 2.4 Toi uu hda cdc yeu td dnh hirdng den hiiu qud ket lu sinh hgc Tiln,hanh khdo sit anh hudng cua pH mdi tmdng, cdc mudi kim loai bfi sung, mfii trudng nufii cay, lieu lugng vi khudn den hieu qud kit ty sinh hpc Tir dd xic dinh dupe dieu kiln thich hgp Bang 1: Cac nguon carbon, nitrogen va mudi kfaoang diing dl nudi vi khudn Nguon nitrogen Ngufin khoang Acid Glutamate (5%) KCl (0.5%) FeCb (0.5%) Yeast exttact (0.05%) CaCb (0.5%) Urea (0.05%) 0.2% K3HPO4 + 0.5% KH2PO4 (NH4)2S04 (0.05%) 2 Ung dung vi khudn sdn xudt chdt ket tu 3.1 Kit qua phan Igp vi kfauin sinh hgc vdo xu ly nudc ri rdc Tir 10 mau nudc ri rac phan lip dugc 55 ddng vi kfaudn, ttong dd: 19 dong vi khuan phan lap tir Ap dyng cdc dieu kien thich hpp da tim dugc mfii truong polysaccharide (chiem 34,55%) vi 36 cho dong vi khuan dugc tuyln chpn dem xii ly dong vi khudn tir phan lip ttr mfii trudng protein nu'dc ri rac quy mfi phong thi nghiim (binh lit) (chiim 65,45%) Qui trinh nhu sau: cho vdo binh cd the tich 10 lit chiia lit nudc ri rdc, bfi sung khoing kim loai, Da sd cic ddng vi khuan co dgng tchuan lac dung dich vi khuan, dilu chinh pH theo cdc gia tti tron, mau frdng, cd bi mdt fron, nhiy va udt, chinh tdi uu da tim dugc; khuay 200 vdng/phiit dp nhdy vi udt tdm khuan tac co die diim kfaac phut, dl yin 30 phiit Lay phdn chi lieu nhu: khudn Igc mfi cao rdn chdc (Hinh IA), TSS vi COD khuin tac udt nfaiiu va chay lan xung quanh (Hinh IC), dgng que ngdn, kicfa thudc tir 0,8 xl,25 pm KET QUA VA THAO LU^N (Hinh iC) Ngudn carbon Glucose (1%) Sucrose (I %) Tinh hot (1%) Hinh 1: Dgc diim mdt so dgng khudn lac eua vi khuin va hinh dang vi khuan chup d kinfa hien vi diln tir quet vdi d$ phfing dgi 7.500 lan ttudng polysaccharide rat thip ( 50% (chiim dugc chpn dl tfii uu hda cac dieu kiln thich 27.27%) 40 dong cd ty II kit tu < 50% (chilm ti 11 hgp chofaieuqua kit ty cao nhdt (Hinh 2) 72.73%) Cac ddng vi khuin cd ti 11 kit tu mdi p chi Kboa hgc Tru&ng Dai hgc Can Tho Phdn A: Kboa hgc Tunhien Car^ nghe id Mdi mtang: 28 i20l3i '>6-92 P: dong vi khuin cd nguon gdc protein Ps: dong vi khuin co ngudn goc polysaccharide PZ.3 FS7.3 Pi Hinh 2: Miroi lam dong vi kfaudn ed ti 11 k i t ta (%) Idn hon % 3.3 Nhan dien \ i kbuan d d n g P l l va P2.4 b a n g phinmg pfaap sinh hgc phan tir Klebsiella sp N B R C 100048 \k FJ577970 Klebsiella sp T-6-1 miic 9 % vd ddng P2.4 t u a n g ddng d miic % vdi ddng JX021670 Enterobacter sp A vd ddng JN695719 Enterobacter sp TX2 (2011) dugc xac dinfa frln cdy p h i he (Hinh 4) cfaia thinfa nfaanfa rd r e t Tat c i ddng vi khudn d i u dugc nhdn diln d bdng 1500 bp frln diln di ctia PCR-16S rRNA dugc nhan len tir DNA ciia cfaiing (hinh 3) v i dugc x i c djnh ddng P l l dfing hinfa vdi AB681138 1^ M 1000 bp •iguamm^mmilUk Hinh 3: Pfao dien di ciia san pfaam PCR dirge nhan l l n t u DNA ciia hai ddng vi k h n i n P l l va P2.4 Ghi chii M:thang chudn lOObp 1.2.3: ddng PI 1, 4.5.6: dong P2 7- ddi chimg dm A6G81138_KlEt)£da_5pL_NBRC_10004B Nhanh FJ57797!)_IOetsiela_sfi_T-frl I JX215711_ErtBaaier_so_A2 Nhanh "^3 • JHS95719_Ef(mtecter_so _T3G_l^lj Hinfa 4: Cay pha fae trinh bay moi quan he giua cac dong \\ khuan sdn xnat cbat k i t tn sinb hoc cd nguon goc protein dua trin trinh tu 16S rRNA (phuong phap neighbor-joining) vdi 1000 lap lgi (bootstrap) 89 Tgp chi Khoa hgc Tnr&ng Dgi hgc Cdn Tha Phdn • Khoa hgc Tu nbien Cong nghe va Moi tnr&ng- 28 (2013) 86-92 3.4 Toi mi hoa cdc yeu to anh faudng den hi?u qua kit tu ciia hai ddng vi kfaudn P l l va P2.4 Dfii vdi ddng P l l : Ngufin carbon, nifrogen va khodng vfi co tfii uu diing dk nufii vi khudn cho ti 11 kit ly cao nhdt (82.4%) la tinh bpt, yeast vd CaCh (Bdng 2); pH tfii uu la (93,96%) (Hinfa 5); mufii kim logi bfi sung cho ti 1| kit ty cao nhit la CaCb (90,08%) (Hinh 6) Dfii vdi ddng P2.4: Ngudn carbon, nitrogen vd khoang vd co tdi uu diing dl nuoi vi khuan cho fr' Ie kit tu cao nhit (81,28%) Id sucrose, NH4CI va K2HPO4 + KH2PO4 (Bang 2); pH tfii uu la (90,57%) (Hinfa 5); mufii kim logi bfi sung cho ti 11 kit ty cao nhdt la NaCl (93,60%) (Hinh 6) chi sau 30 phiit Hinh 5: Anh hudng cua pH den hi^u qua kit tu ciia ddng vi khudn P l l vd P2.4 Bang 2: Anh faudng ciia nguon carbon, nitrogen vii kfaoang vfi co din kha nang kit tu sinh hpc cua bai dong vi khuan P l l va P2.4 Dong (1%) Pl.l LSD (5%) 4.14 g (1%) Tinh bOt (1%) Glucose (1%) LSD (5%) 3.95 Sucrose (1%) (1%) Glutamate (5%) 71,95 36.89 52.57 Nguon nitrogen Yeast Ure (0,05%) (0,05%) 47.42 54,26 51.34 39,88 48.24 49.00 NHiCl (0,05%) 26,21 10.19 18,92 Nguon khoang vo CO* KCl (0,5%) FeClj (0.5%) C a C b (0,5%) K2HPO4 (0,5%) + K H P O , (0,2%) KCl (0,5%) FeCls (0,5%) CaCI, (0,5%) K:HPO4(0,5%) + KH0PO4 (0,2%) KCl (0,5%) 35,70 32,70 58.66 27,42 37,70 32,63 49.13 50,24 44.96 72,73 40,58 21,52 66,74 53,04 40.53 54,80 52,47 44,65 57,96 56,23 61,63 47.00 70.89 64.30 56.98 82,40 75.03 61,02 58,57 52,51 39,93 77,92 48,61 62,68 44,82 65,45 58,48 41.79 43.1 40,1 42,41 48,03 42,98 52.31 44,73 12,63 12,99 22,81 36.68 38.66 56.48 23,58 49,44 29,89 46.58 41,59 80,86 47.36 43,8 76.7 48.58 52,60 32,61 57,84 67.79 42.58 64.17 81,28 57,90 59.01 62.73 49,32 78,42 77,08 KIHP04 (0,5%) + KH2PO4(0,2%) KCl (0,5%) 60,89 75,31 F e C b (0,5%) 64.05 69.62 77,70 78,93 63.65 62.95 78,73 78,89 C a C b (0,5%) K2HPO4 (0,5%) + KH2PO4 (0,2%) FeClj (0.5%) CaCl2 (0,5%) K2HPO4 (0,5%) + KH,P04 (0,2%) KCl (0,5%) FeCl, (0,5%) C a C b (0,5%) K2HPO4(0,5%) + KH2PO4 (0,2%) KCl (0,5%) F e d ) (0,5%) C a C h (0,5%) Tap chi Khoa hgc Tnrang Dai hgc Cdn Tha Hinh 6: Anh huong cua muoi kim logi den failu qua kit tu ciia ddng vi kfauin Pl.l va P2.4 PhdnA: Khoa hgc Tu nhien Cong nghe va Mdi truang 28 i2(il3r >^6-9- c »*• t UM ^ „^ ; ^ faai deu cd nang xir li nudc thdi tinh bfit, lim giam 95,85% COD vd 66,5% TSS va nudc thii protein la 90,43% COD vi 50% TSS sau 30 phiit ^h ling so voi dfii chiing (Cao Ngpc Diep et al 2010) Biii Thi Vinfa et al (2010) phin lip, tuyin chpn hai ddng vi kfaudn Enterobacter aerogenes Pll va P14 tir chit thai nfad may siia vd frai chdn nufii bd siia, hai ddng cd kha ndng xii ly nudc thai sita vd nudc ri rac frong lim gidm ham lugng TSS va COD lln din 43,6% vd 39%; 63,9% vi 67% theo thu ty sau 30 phiit dl ling; ket qui ciia chiing ldi cung nhin thay hai ddng vi khuin lao chdt kit ty sinh hgc Enterobacter sp P2.4 (ngudn gfic protein) cung cd khd ndng lam giim 21,9% COD va 12,4% TSS ciia nudc ri rac cimg thoi gian, sd di cd sy khdc bill giiia cdc dong vi khuin cd ti II gidm lugng COD vi TSS d cac logi nudc thii khae Ii vi d nude thdi tinh bpt (xudng san xuat biin) chi cd linh bfit Id chdt la lung tao nen nudc thii dye nen hi|y qua vi khuin tgo chdt kit ty sinh hpc lim gidm lugng TSS va COD cao so voi logi nudc thdi cdn lai, chiia nhilu tap chat hon vi cac kit qua niy gdp phan vao xii ly cfaung nudc ri ric Trong Bing trinfa biy sy tfi hgp cic ngufin dinh duong (carbon, nita, khoang) de co ti 11 ket tu cao nhit nhu dong Pl.l voi 1% tinh bpt, 0,05% yeast extract, 0,5% CaCb cho tl le kit lu cao nhat (82.4%) tuong ty ddng P2.4 li 1% sucrose, 0,05% NH4CI, K2HPO4 (0,5%) + KH2PO4 (0,2%) cho ti ll kit ty cao nfait la 81.28% 3.5 iTng dung ddng vi khuin Pl.l va P2.4 viio xir Iy nudc ri rac Kit qua xu ly nudc ri rdc cho thay ca hai dong Pl.l va P2.4 lam giam hdm lupng TSS khdng khdc bi|t (12,09% vdi dong Pl.l va 12,4% vdi ddng P2.4); Dong P2.4 lam giam lugng COD tfit hem ddng Pl.l (21,89% so vdi 19.84%); Khi phfii hgp ca hai ddng lai thi hieu qua xir ly lugng TSS tfit hon (gidm 26.37%)(Bing 3) Tir 20 miu nudc thai dugc thu d linh Tiln Giang, chiing toi phin Igp dugc 53 dong vi khuan sin xuat chat kit tu sinh hpc dfi tuyen chpn vi nhan diln dugc ddng vi khudn Cronobacter sakazakii T2a (c6 nguon gdc tir mdi tnrong polysaccharide) va dong Cronobacter sakazakii PI lb (co ngufin gfic tir mfii tnrong protein) va ca Bang 3: Ket qua xii ly nudc ri rac ciia dong vi kfaudn P l l va P2.4 Chi ti£u COD, mg 0/lit TSS mK/m Triro-c xirlv Pl.l Sau xirlv Till Siam (%) Triro-c xiJK 1270 1018 19,84 1270 637 560 12.09 637 KET LUAN Hai dong vi khuin Pl.l va P2.4 phan Igp tir nudc ri rac cho ti le kit ty vdi dung djch kaolin bd sung CaCh ldn lugt la 85.81% vi 84.24% va hai ddng Pl.l cfi ti 11 dfing hinh gen I6S rDNA lin lugt vdi AB681138 Klebsiella sp NBRC 100048 va FJ577970 Klebsiella sp T-6-1 (99%) vi dong P2.4 Tile Sau xirlv eiam (%) 992 21,89 P1.1 + P2.4 Trir*c Sau Ti le xii Iy xii ly giam (%) 1270 1028 19,06 12,40 558 637 469 26,37 P2.4 vdi JX021670 Enterobacter sp A2 va rN695719 Enterobacter sp TX2 (2011)(98%) - Cdc dilu kiln toi uu cho hieu qua kit tu cao nhdt gfim: pH (ddngPl.l), pH (dong P2.4); ion khoang bo sung Id 0,5% CaCb (ddng PI 1) va 0,5% NaCl (ddng P2.4); ngudn carbon, nifrogen va khodng vfi ca cho moi frudng nudi ciy lin lugt la 1% tinh bfit 0.05% yeast extract vi 0,055 CaCb Tgp chi Khoa hgc Tnrang Dgi bgc Cdn Tho PhdnA Kboa hoc Tu nhien Cong nglie vd Moi iru&ng 2812013 r 86-92 (ddng PI 1) va I % sucrose, 0.05% NHjCI vi 0,5% K2HPO4 + 0,2% KH2PO4 hidl CAM T ^ Dl tdi dugc thyc hien vdi sy hd frg kinh phi cua Sd Khoa hpc vi Cfing ngfal Thanh phd TAI LIf U THAM KHAO Bui Thi Vinfa, Phan Thanh Quoc vd Cao Ngpc Di|p 2010 Phin lap vd nhdn diln vi khuan sdn xudt chdt kel tu sinh hpc frong chat thii sira vd img dung frong xii ly nudc thai Tap chi Cfing nghe Sinh hoc (3A): 805-809 Cao Ngpc Dilp, Ll Thi Loan vi Trin Ngpc Nguyin 2010 Phin ldp vi nhin diln vi khudn san xudt chat kit tu sinh hpc va ung dyng xir \y nudc thai Tap chi Cflng nghi Sinh hpc 8(2): 253-264 Deng S B., R Bai and X Hu 2002 Characteristics of a bioflocculant produced by Bacillus mucilaginosus and its use in starch wastewater treatment Appl Microbial Biotechno 60: 588-593 Gong W.X., S.G, Wang., X.F Sun., X.W Liu., Q Y Yue and B.Y Gao 2008 Bioflocculant production by culture of Serratia ficaria and its application in wastewater treatment Bioresour Technol 99; 4668-4674 Hazana R and Norli 2008 Flocculanting activity of bioflocculant producing bacteria isolated from Closed drainage system at Prai industrial Zone Penang, Malaysia International conference on Enviromental _ Reseach and Technology, pp 422-425 Kim, J.M., H J Lee S Y Kim, J J Song, W Park, and C O Jeon 2010 Analysis of the Fine-Scale Population Structure of "Candidatus Accumulibacter phosphatls" in Enhanced Biological Phosphorus Removal Sludge, Using Fluorescence In Situ Hybridization and Flow Cytometric Sorting Applied and Environmental Microbiology, 76(12): 3825-3835 Levy N., S Magdass and Y Bar-Or 1992 Physico-chemical aspects in floccuiation of bentonite suspensions by a cyanobacterial bioflocculant Water Res., 26: 249-254 Lu W Y T Zhang , D Y Zhang , C H Li., J P Wen and L X Du 2005 A novel bioflocculant produced by Enterobacter aerogenes and its use in defecating the trona suspension J Biochem Eng.,27: 1-7 9- Salehizadeh H., M Vossoughi and I Alemzadeh 2000 Some investigations on biofloccutant-produemg bacteria Biol Chem., 5: 39-44 10 Sheng Y., Q Zang and H Wang 2006 Sreening and flocculating of bioflocculant produccing microorganisms Sience and Technology Beijing, 13:289 11 Shifa I L., Y.T Van., L C Yeh., H G Lin and Y.N Chang 2001 Production of a biopolymer flocculani from Bacillus licheniformis and its floccuiation properties Bioresour Technol., 78: 267-272 12 Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., and Kumar, S., 2011 MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likehood, Evolutionary Distance and Maximum Parsimony Methods Moi Biol Evol., 28: 2731-2739 13 Yim J.H., S.J Kim., S.H Ahn and H.K Lee 2007 Characterization of a novel bioflocculant, p-KG03, from a marine dino agellate Gyrodinium impudicum KG03 Bioresour Technol., 98:361-367 14 Yokoi H., O Natsuda and J Hirose 1994 Characteristics of a bio-polymer flocculani produced by Bacillus sp PY-90 Ferment Bioeng., 79:378-380 15 Zang J., Z Liu, S Wang, P Jiang 2002 Characterization of a bioflocculant produced by the marine myxobacterium Nannocystis sp NU-2 Appl Microbiol Biotechnol., 59:517-522 16 Zhi-qiang Z., L Bo., X Si-qing, W Xuejiang and Y A-ming 2007 Production and application of a novel bioflocculant by multiple-microorganism consortia using brewery wastewater as carbon source Joumal of Environmental Sciences, 19: 667-673 17 Zouboulis A 1., X L Chai and l.A Katsoyiannis 2004 The application of bioflocculant for the removal of humic acids from stabilized landfill leachates J Environ Manage., 70: 35-41 ... tl vi chiing khd bj phin hu>'' ttong tu nhien (Yokoi el al., 1994) Gin da\ kit ty sinh hpc bdi vi sinh vat dugc ngdnh cdng ngfal sinh hpc mdi tmdng chii y bdi vi chiing cd khd ndng pfaan gidi sinh. .. GIOI THIEU Kit tu sinh hgc Id sdn phim ciia hgp chdt cao phin tir dugc hmfa thdnh frong qud trinh ph^ trien cua vi sinh vit (Shih et al., 2001) Vi khuin san xuit chit kit ty sinh hgc Id nhiing... nghe Sinh hoc (3A): 805-809 Cao Ngpc Dilp, Ll Thi Loan vi Trin Ngpc Nguyin 2010 Phin ldp vi nhin diln vi khudn san xudt chat kit tu sinh hpc va ung dyng xir \y nudc thai Tap chi Cflng nghi Sinh