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[.]
Trang 1Tap 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 6 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 can flocculate and settle dispersed organic particles so thai they can be used for wastewater treatment and it is not toxic to human and the environment Fifty-five bioflocculant-producing bacterial isolates were isolated from ten leacheate samples of five landfills 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 FJ577970 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 consists
of starch YE CaCh for Pl.l isolate: sucrose, NH,Cl K2HPO.i+KH2POjfor P2.4 isolate with kaolin solution after 5 minutes at pH=4 (Pl.l isolate) and pH=5 (P2 4 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 6 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 con 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 5 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 trong 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 88.58%: ddng P2.4 Id sucrose, NH4CI, K2HP04+KH2PO.I cho fi H kit tu Id 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 trong 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)
Trang 21 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
dudi sy hogt ddng ciia cde enzjme die biet, sau dd
chiing ed tfae dugc bdi tilt ra ngodi vd Idn tgi frong
mdi tmdng faoac frln bi mdt vd te bao vi khudn,
cfait khde nhau vd tgo thanh 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\ ra 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 ra 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
nhanh chdng vi an toan khdng ddc hai cho con
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
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 ciia faai dong vi
khuin cd ti le kit ty cao (> 80%) va img dung
chiing trong xu ly nudc ri rdc die biet tara gidm
ham tugng COD va TSS frong thinh pfaan nudc
ri rae
2 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 do pha lodng hiit 20 pi trdi diu trin moi trudng thach, sau dd li 2 — 3 ngay 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 tiln fainfa cay cfauyin sang dng Kl tiep (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 trong 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 4 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 5 phiit sau do liy phin ttong d tten dem do 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
Trang 3diln di sdn p h ^ PCR dugc cfayp fainh qua GelDoc
vdi thang cfaudn 100 bp sau do chiing dugc gidi
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
cfaung bing phdn mim MEGA 5.05 (Tamura et ai,
2011) voi phuong phdp neighbor-joining dya tren
1000 Idn lap lai (bootsttaps)
2 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
cfao faieu qua 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 thanh phan frong Bing 1, ta dugc 48 nghiim thiic khac nhau ve mfii tmong nufii vi khudn Nufii vi khuan ttong dng nghiem loai 10 ml d nhiet dp phdng, lie 160 vdng/pfaiit Sau 4 ngay, kiim tra hi?u qud kit ly sinh hpc vdi Kaolin
diing dl nudi vi khudn
Glucose (1%)
Sucrose (I %)
Tinh hot (1%)
Acid Glutamate (5%) Yeast exttact (0.05%) Urea (0.05%) (NH4)2S04 (0.05%)
KCl (0.5%) FeCb (0.5%) CaCb (0.5%) 0.2% K3HPO4 + 0.5% KH2PO4
2 2 5 Ung dung vi khudn sdn xudt chdt ket tu
sinh hgc vdo xu ly nudc ri rdc
Ap dyng cdc dieu kien thich hpp da tim dugc
cho dong vi khuan dugc tuyln chpn dem xii ly
nu'dc ri rac quy mfi phong thi nghiim (binh 5 lit)
Qui trinh nhu sau: cho vdo binh cd the tich 10 lit
chiia 5 lit nudc ri rdc, bfi sung khoing kim loai,
dung dich vi khuan, dilu chinh pH theo cdc gia tti
tdi uu da tim dugc; khuay 200 vdng/phiit trong 2
TSS vi COD
3 KET QUA VA THAO LU^N
3.1 Kit qua phan Igp vi kfauin
Tir 10 mau nudc ri rac phan lip dugc 55 ddng
vi kfaudn, ttong dd: 19 dong vi khuan phan lap tir mfii truong polysaccharide (chiem 34,55%) vi 36 dong vi khudn tir phan lip ttr mfii trudng protein (chiim 65,45%)
Da sd cic ddng vi khuan co dgng tchuan lac tron, mau frdng, cd bi mdt fron, nhiy va udt, chinh
dp nhdy vi udt tdm khuan tac co die diim kfaac khuin tac udt nfaiiu va chay lan xung quanh (Hinh IC), dgng que ngdn, kicfa thudc tir 0,8 xl,25 pm (Hinh iC)
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 3.2 Hiiu qua ket tu ciia cac dfing vi khuin
phin Igp
Kit qua ghi nhin frong 55 ddng vi khuan da
phin lap cd 15 ddng ed ti I? kit tu > 50% (chiim
27.27%) 40 dong cd ty II kit tu < 50% (chilm ti 11
ttudng polysaccharide rat thip (<50%) frong khi dd cac dong vi khuin phan lap tir mdi ttudng protein
cd ddng Pl.l cfi ti le kit ty cao nhdt (85.81%) rdi din ddng P2.4 (84,24%) vi hai ddng Pl.l vd P2.4 nay dugc chpn dl tfii uu hda cac dieu kiln thich hgp cho faieu qua kit ty cao nhdt (Hinh 2)
Trang 4P: dong vi khuin cd nguon gdc protein Ps: dong vi khuin co ngudn goc polysaccharide
PZ.3 FS7.3 Pi 2
Hinh 2: Miroi lam dong vi kfaudn ed ti 11 k i t ta (%) Idn h o n 5 0 %
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
Tat c i 2 ddng vi khudn d i u dugc nhdn d i l n d
bdng 1500 bp frln pho d i l n di ctia PCR-16S rRNA
x i c djnh ddng P l l dfing hinfa vdi AB681138
M 1 2
1^
Klebsiella sp N B R C 100048 \k FJ577970 Klebsiella sp T-6-1 miic d o 9 9 % vd ddng P2.4
t u a n g ddng d miic 9 8 % vdi ddng JX021670
Enterobacter sp A 2 vd d d n g JN695719 Enterobacter sp T X 2 (2011) d u g c xac dinfa frln
cdy p h i he (Hinh 4) cfaia thinfa 2 nfaanfa rd r e t
5 6
1 0 0 0 bp
•iguamm^mmilUk
Hinh 3 : Pfao dien di ciia san pfaam P C R dirge nhan l l n t u D N A 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 4 7- ddi chimg dm
A6G81138_KlEt)£da_5pL_NBRC_10004B FJ57797!)_IOetsiela_sfi_T-frl
I JX215711_ErtBaaier_so_A2
"^3 • JHS95719_Ef(mtecter_so _T3G_l^lj
Nhanh 1
Nhanh 2
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 t r i n trinh t u 16S rRNA ( p h u o n g phap neighbor-joining) vdi 1000 lap lgi
(bootstrap)
Trang 53.4 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 4 (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
nuoi vi khuan cho fr' Ie kit tu cao nhit (81,28%) Id
sucrose, NH4CI va K2HPO4 + KH2PO4 (Bang 2);
sung cho ti 11 kit ty cao nhdt la NaCl (93,60%)
(Hinh 6) chi sau 30 phiit
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
Nguon nitrogen
Hinh 5: Anh hudng cua pH den hi^u qua kit tu ciia 2 ddng vi khudn P l l vd P2.4
Dong
P l l
L S D
(5%)
4.14
LSD
(5%)
3.95
(1%)
g
(1%)
Tinh bOt
(1%)
Glucose
(1%)
Sucrose
(1%)
(1%)
Glutamate
(5%) 71,95
36.89 52.57 35,70 37,70 32,63 49.13 52,47 61,63 47.00 70.89 48,61
58,48
41.79 42,98 36.68 49,44 29,89 80,86 67.79 57,90 59.01 64.05 63.65
Yeast
( 0 , 0 5 % ) 47.42 51.34 48.24 32,70 50,24 44.96
72,73
44,65 64.30 56.98
82,40
62,68 43.1 40,1 52.31 38.66 47.36 43,8 76.7 42.58 62.73 49,32 69.62 62.95
Ure
( 0 , 0 5 % ) 54,26 39,88 49.00 58.66 40,58 21,52 66,74 57,96 75.03 61,02 58,57 44,82 42,41 48,03 44,73 56.48 46.58 48.58 52,60 64.17 78,42 60,89 77,70 78,73
NHiCl (0,05%)
26,21 10.19 18,92 27,42 53,04 40.53 54,80 56,23 52,51 39,93 77,92 65,45 12,63 12,99 22,81 23,58 41,59 32,61 57,84
81,28
77,08 75,31
78,93
78,89
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%) FeClj (0.5%) CaCl2 (0,5%) K2HPO4 (0,5%) +
K H , P 0 4 (0,2%) KCl (0,5%)
F e C l , (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%)
K I H P 0 4 (0,5%) + KH2PO4(0,2%) KCl (0,5%)
F e C b (0,5%)
C a C b (0,5%) K2HPO4 (0,5%) + KH2PO4 (0,2%)
Trang 6Hinh 6: Anh huong cua c »*•
muoi kim logi den failu t UM
qua kit tu ciia 2 ddng vi ^ „ ^ ;
kfauin Pl.l va P2.4 ^
Trong Bing 2 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 2 ddng vi khuin Pl.l va P2.4
viio trong 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 trong dfi tuyen chpn
vi nhan diln dugc 2 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 2 dong vi kfaudn P l l va P2.4
faai deu cd khi 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 do lim gidm ham lugng 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 trong 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 nhau 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 2 logi nudc thdi cdn lai, chiia nhilu tap chat hon vi cac kit qua niy gdp phan vao xii ly cfaung nudc ri ric
Chi ti£u
COD, mg
0/lit
TSS mK/m
Triro-c
xirlv
1270
637
Pl.l Sau xirlv
1018
560
T i l l Siam (%) 19,84 12.09
Triro-c xiJK
1270
637
P2.4 Sau xirlv
992
558
Tile eiam (%) 21,89 12,40
P1.1 + P2.4 Trir*c Sau Ti le xii Iy xii ly giam (%)
1270 1028 19,06
637 469 26,37
4 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 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 4 (ddngPl.l), pH 5 (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
Trang 7(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
1 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 8 (3A):
805-809
2 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 trong xir \y nudc thai Tap chi Cflng
nghi Sinh hpc 8(2): 253-264
3 Deng S B., R Bai and X Hu 2002
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