Tgp chi Khoa hgc Trudng Dgi hgc Cdn Tho Phdn A: Khoa hgc T^ nhien, Cdng nghi vd Moi trudng- 33 (2014): 117-124
Tap chl Khoa hoc Tru*6ng Oai hpc Can Thtf website: sj.ctu.edu.vn
P H A N L A P , T U Y E N C H O N V A iTNG D U N G V I K H U A N C H U Y E N H O A N I T O V A P H O T P H O T t r B A I R A C D E XU" L Y N V A P T R O N G NlTOfC R I R A C
Cao Ngoc Didp' va Doan Tin Lyc^
' Vien Nghien cuu & Phdt trien Cong nghe Sinh hoc. Tnrdng Dgi hgc Cdn Tho
^ Trudng Trung hoc Phd thong VTnh Xudn, Trd On, Vinh Long
Thdng tin chung:
Ngdy nhdn: 24/03/2014 Ngdy chdp nhdn: 28/08/2014
TUle:
Isolation, Selection and Application of Nitrogen Removal Bacteria and Polyphosphate- Accumulating Bacteria fi-om biowaste station for N and P removal treatment in sewage water
Tirkhda:
Ammonium, nuac ri rdc, orthophosphate, vi khudn chimin hod nita. vi khudn tich luy pophosphate
Keywords:
Ammonium, nitrogen removal
bacteria.orthophosphale.
polyphosphate- accumulating bacteria, sewage water
ABSTRACT
Forty-five nitrogen removal bacterial isolates and fifiy-tivo polyphosphate- accumulaling bacterial isolates were isolated from sewage water of five biowaste stations (Vinh Long. Can Tho, Hau Giang). After testing on minimum medium added NH4*, NOf, NO3' and mixture of these three kinds of nitrogen anh orthophosphate, three isolates having high N removal ability (TODLl. TOD2 3, TND1.4) and three polyphosphale-accumulating bacterial isolates (TL2 3. HP2.3, HP3.2) were chosen to sequence randomly by automatic sequencer. DNA sequencing were compared with GenBank databank ofNCBI by BLAST N soft»'ore. The results showed that TODl. 1, TOD2.3 and TND1.4 isolates were 99% of identity with JF799886 Enterobacter sp CIFRl D-TSB-9-7MA, HQ259961 Klebsiella variicola strain 7 & EU884439 Enterobacter sp. 12 and FJ189785 Enterobacter sp. CSB08. respectively; while polyphosphale-accumulating bacterial TL2.3 isolate had the identity rale of 98%
with JX025736 Bacillus cereus strain VPll, the HP2.3 and HP3.2 isolates were 99% of identity with JF505965 Exiguobacterium mexicanum strain KNUC9031 and FJ976560 Acinelobacter soli strain LCR52, respectively. The combined Enterobacter sp. TNDl.4 and Acinelobacter soli HP3.2 reduced the concentrations of ammonium, total N, Nitrite, Nitrate, orthophosphate, total P and the released ammoniac gas lo the lowest level after five days of inoculation.
TOM TAT
Bon muai lam dong- vi- kJmSh chuyin hod nito vd 52 ddng vi khudn tich luy polyphosphate dugc phdn Igp lir n udc ri rdc ciia 5 bai rdc d V inh Long, CSn Thavd Hgu Giang. Khdo sdt trin mdi trudng bd sung NH4*, NO2', NOf vd hon hgp 3 logi Niia (bag gdm NH4", NO2', NO3) vdi lugng orthophosphate, 3 ddng vi khudn chuyin hda niia (TODLl, TOD2.3, TND1.4 ) va 3 ddng vi khudn tich luy poly-P (TL2.3, HP2.3. HP3.2) cao dugc chgn de gidi irinh lu vd su dung phdn mim BLAST Ndi so sdnh vdi cdc trinh tir cdc ddng vi khudn trin ngdn hdng du lieu gen ciia NCBL Kit qud cho ihay dong TODLl ddng hinh vdi JF799886Enlerobgclersp. CIFRID-TSB- 9-ZMA vd ddng TOD2.3 tuong dong vdi dong HQ259961 Klebsiella variicola strain 7 & ddng EU884439 Enterobacter sp. 12; ddng TND1.4 ddng hinh vdi ddng FJ189785 Enterobacter sp.CSB08 diu d muc 99%; Irong khi ddng vi khudn lich luy poly-P TL2.3 tuang ddng d mdc 98% vdi ddng JX025736 Bacillus cereus strain
VPll vd hai ddng HP2.3 &HP3.2 ddng hinh 0 miic do 99% vdi ddng JF505965 Exisuobaclerium mexicanum strain KNUC9031 vd ddng FJ976560 Acinelobacter soli strain LCR52, theo ihutu. Hon hgp hai ddng Enterobacter sp. TND1.4 vd Acinelobacter soli HP3.2 gidm lugng ammonium, TN, Nitrite, Nitrate, orthophosphate vd TP trong nude ri rdc a muc ihSp nhdl sau 5 ngdy sau khi chung ddng thdi hdm lugng khi ammoniac thai ra Ihdp nhdl.
Tgp chi Khoa hgc Trudng Dgi hgc Cdn Tho Phdn A: Khoa hgc Tu nhien, Cdng nghi vd Mdi trirdng: 33 (2014): 117-129
I ©^T VAN BE
Cftng vdi tic dp do toi hod, cdng nghifp hoa nhanh chdng din den sy d nhiem vi suy tooii nghiem trpng mdi trudng, dang chu y li d nhilm mdi trudng nude. Nude khdng chi bi d nbidm bdi nude toil cdng nghifp, nude toil sinh hogt, sft dyng qua nhilu phin bdn hda hgc uong sin xuit ndng nghifp... mi con hi d nbidm bdi nhieu ngudn khdc trong do, ding quan tam nhit l i nude ri ra tft cie bai rdc vi logi nude nay cd ham lupng chit d nhilm rit cao. Thdnh pbin nude ri rie rit phftc tap tiong do d nhilm chat hftu ca la chu yeu, ben cgnh cdn 6 nhilm chit vd ca, die bift li mgt lupng ldn cic hpp chit nito vd ca boa tan (NH4*, NO3", NO2"'.
Ndng dd NH4 cao giy hien tupng phu dudng lim mdi Uudng nude d nhiem nang v i hdi thdi. Ben canh dd, thanh phin lan boa tan chiem mpt ^ If Idn cung gdp phin to ldn giy 6 nhilm mdi trudng.
Lupng nude ndy nlu khdng dupe xft ly dung mftc thi ed nguy co giy d nhiem cae ting nude mat Ud thdnh nguydn nhdn tryc tilp phat sinh dich bfnh, giy bfnh cho con ngudi va inh hudng din mdi trudng xung quanh. Ddng thdi, nd co tod xdm nhap vio mdi Uudng dit sau do di vao cae mach nude ngim lam d nhilm nguin nude ngim vd lam biln ddi die tinh cua dit. Tuy nhien, Uong nude ri ric cung chfta nliidu chung logi vl khuan chuyen hoi nita, vi khuin tich luj polyphosphate... rat phong phft vd chinh nhung logi vi khuin bin dia ndy nlu dupe tuyln chgn va ftng dyng de xft li N va P hoa tan trong nude ri rdc mang lai hifu qua cao. Muc tieu cfta nghien cftu niy la phin lip, tuyen chgn vi ling dyng cic ddng vi khuin chuyen hod nita, vi khuin tich luj polyphosphate tdt de loai bo.
2 PHUONG PHAP NGHIEN CtTU 2.1 'V^t lifu
- Miu dupe tou d ddu ra cfta cdc bii rdc Hoa Phft, Tri On (VTnh Long), Tdn Long (Phung Hifp, Hgu Giang) va Thdt Ndt, Cd Do (Thdnh phd cin Tha).
Mau dupe tou bang each: trdng lp nhya bing nude toil liy dupe tu dng toai, sau dd liy nude toil rdi day nip, ghi nhan, dgt lp vao bpc nilon rdi cho vao toiJng tru Ignh de mang ve phong thi nghiem vi bio qudn.
2.2 Phinmg phip nghien cuu 2.2.1 Phdn lap vi khudn
Dung micropipet hftt 50 pi mdu nude (d dp pha loing thich hpp) rii trai len dia mdi trudng phan Igp. Dung que Uii mau phan phii djch mlu trdi deu
khip mat thach, ft Uong tu ft vi sinh d nhiet df 30''C Uong ttidi gian 48 gid (mdi tiordng cho vi khudn ehuyin hda nito (Zhao et al., 2010) va 72 gid (mdi uucmg cho vi khudn ti'ch luy poly-P ( W ^ g et al., 2008). Khi vi khuin phit ft-iln toanh khuin lac, chpn eic khuin lgc rdi rgc vi cd hinh toai khdc nhau Uen mdi trudng phan Igp (ye hinh dgn^ mdu sic, kich toudc, bia, dp nil) moi dgng khudn lgc ciy chuyin nhilu lin sang mdi trudng Cling loai den khi cic khuin Igc rdi ra, ding nhat va diu nhau Uen dudng ciy. Chpn khuin lac rdi cay chuydn vio ing chfta mdi trudng cung logi. 6ng di ciy chuyin duge ft d 30°C ttong tu ft vi sinh vgt dl vi khuin phit Uiln. Sau dd dem trft Ignh dl chd kilm Ua do rdng dudi kinh hidn vi.
2.2.2 Tityen chgn cdc ddng vi khudnph^ l^ dugc Kiim tra khd ndng chuyen hoa nltff cua cdc dong vi khudn
Cac dong vi khuin phin Igp dupe ciy trdn 4 loai moi trudng cd b i sung NH4*, NOa', NO2" vi T (NH4 , NO3, NO2) trong dd NH4 v i NO3 d ning dg 100 mM, NO2" d ning dd 10 mM dl so sanh khd nang phat Uiln cfta chftng. Chgn nhftng ddng vi khuan phit Uiln dupe Udn mdi trudng c6 bi sung NH4 , NO3 , NO2 va T (NH4+, NOs", NOaO ftong dd NH4'' v i NO d ning df 100 inM, N02' d ning dd 10 mM dl tilp tye kilm tra khd ndng phit triln cfta chftng Udn mdi Uudng cd bi sung NH4^, NO3", NO2 vd T (NXi4 , NO3, NO2) 6 ning dg cao ban Uong dd NH4 v i NO3' d ning df ting din 200 mM, 300 mM... v i N02" d ndng df ting din 20 mM, 30 mM...
Kiem tra khd ndng tich luy PO4 cua cdc dong vi khudn tich luy poly-P
Cic dong vi khuin sau khi dupe kilm tra rdng tiln hinh nudi udn mdi trudng Idng trdn miy lie dl kilm Ua khi nang tich luy poly-P. Mdi ddng vi khuin dupe chftng vio 5ml mdi trudng sau thdi gian 4-6 ngay.
Ly trich poly-P (Eixler et al., 2005) bing cich nudi vi khuin Uong chai amplcillin chfta 4 ml mdi tiordng tich luy poly-P, lie 160 vdng/phut, sau 6 ngay nudi tiln hinh ly tim 12.000 v6ng/phftt vi Uong thdi gian 10 phut de tou sinh khoi, logi bd phin nude djch. Hda tan sinh khii trong 4 ml NaOH 0,2 M, ft Uong 20 ^\d de trich poly-P. Djch t l bio dupe lpc qua ^ay Ipc Sartorius Stedim 17598 0,45 pm, CE dl logi bd xic tl bio dl thu dich Ipc chfta poly-P. Dich Ipc thu dupe chia lam 2 phin:
Tgp chi Khoa hgc Trudng Dgi hgc CSn Tha PhSn A: Khoa hgc Tu nhiin, Cdng nghi vd Mdi Irudng: 33 (2014): 117-124 - Phin 1: do dl xic dinh lupng phosphate ty
do (PPl) Uong djch bing phuang phip Molypdate blue (so miu d budc sdng 880 nm Udn mdy Beckman Coulter DU640B).
Phin 2: touy phin bdng HCI IM d 100°C Uong tiidi gian 10 phftt. Dich tofty phin sau do de ngudi va dem do ham lupng phosphate (PP2) vdi phuong phdp ^ i n g nhu phin 1. Ham lupng poly-P ndi bio dupe tinh bing hidu s i gifta PP2-PP1.
Nhihig ddng vi khuin cd dp hftu hifu cao (chuyin hod dam vi tich luj phospho boa tan cao) dupe chpn de nhdn difn bang each thyc hifn cdc phdn ftng PCR dl xic dinh gen 16S rRNA toeo cap mdi 8F v i 1492R (cho vi khudn chuyin hoi nito) v i 27F vd 1492R (cho vi khuin tich luy poly- P); san phim PCR sS difn di udn gel agarose 0,8%
va phi difn di sdn phim PCR dupe chup hlnb qua GelDoc vdi thang chuin 100 bp sau do chung dupe giii trinh ty d cdng iy MACROGEN Bang 1: Tbinh phin ly hoa tinh trong nurdc ri ric tai
(Hdn Quic), kit qui Uinh ty dupe so sanh vdi ddng vi khuan chuin d ngin hang dft lifu cfta NCBI bang phin mim BLAST N dl nhin difn dong vi khuin dupe giai trinh ty gen I6S rRNA.
Xiy dyng cay pha hf cua chftng bang phan mem MEGA 5.05 (Tamura et ai, 2011) voi phuang phip neighbor-joming dya Udn 1000 lin lap lgi (bootsUaps).
2.2.3 Ung dung cdc dong vi khudn tot trong logi bo N&P trong nude ri rdc d mo hinh binh len men IL
Ket hpp 2 ddng vl khudn chuyen boa N va vi khuan tich luy poly-P trong didu kifn syt khi de logi bd N va P trong nude rl rac (cd todnh phdn li boa tinh Uinh bdy Uong Bing 1) Uong md binh binh Ien men (bioreactor IL) cho ddn khi ham lupng Ammonium v i orthophosphate giim dudi mftc logi Bl cfta QCVN 25 :2009/BTNMT).
bai rac Tin Long, Phung Hiep,Hau Giang pH TKN
Miianang 7,2 351,65 Mua mua 7,8 280,91 Ngudn phdn tich tai PTN Chuyin sd Cdng ngh4 thdnh pho Cdn Tho
NH4+
(mg/L) 275,72 402,02 it va Vi sinh
NO2- (mgft.) 0,02 0,57
NO3- TP (mg/L) (mga,)
0,09 113,32 0,09 30,67 mdi [rudng. Dai hoc Cdn Tha vd Trung
POi'- COD TSS (mg/L) (mgrt.) (mgfl.)
23,22 1090 382 12,61 964 156 tdm Ky thudt vd lfng dung
s i lifu Uung binh dupe phan tich thing ke bing phin mim EXEL 2003 v i so sdnh sy khdc bift cd y nghia thing kS b i i ^ kilm dinh LSD.OI hay Duncan.
3 KET QUA VA THAO LUAN Bin muai lim ddng vi khuin chuyen hod nita va 52 ddng vi khuin tfch luy polyphosphateduj
phan lap tft 20 mau nude ri rdc cua 5 bai rac Hod Phft, Trd On (VTnh Long), Tin Long (Hiu Giang), Thit Nit, Cd Dd (Cin Tha). Hiu hit cdc ddng vi khuan phan lip dupe cd die dilm que ngan vi cd khd nang chuydn ddng, khuin lac co dgng bmh trdn, mau Uing due, bia nguydn, dp ndi md, kich toudc tft 0,4 - 2,5 nun (Hinh I).
Hinh I: Khuin Igc cua 2 ddng vi khuin TODl.l va TL2.3 va hinh dgng efia dong TL23 (chyp hinh & kinh hidn vi difn tir quet vdi do phdng dgi 11.000 Ian)
Tap chi Khoa hgc Trudng Dgi hgc Can Thcr PhSn A: Khoa hgc Tir nhiin, Cdng nghi vd Mdi Irudng: 33 (2014): 117-124 Sau khi ddnh gii qua cac ning dp NH4''^,
NO2", NO2" va hdn hpp 3 loai nito (Bing 2) cung lupng orthophosphate ti'ch luj (Bdng 3), chgn dugc 3 dong vi khuin chuyen hda nito (TODl.l, TOD2.3, TND1.4) va 3 ddng vi khuin tich luy poly-P (TL2.3, HP2.3, HP3.2) ed do hftu Bang 2: Ba ddng vi Idiuan co kha nang chuyen he
hifu cao vi chung ed klii ning phit tridn tdt ttdn moi tiirdng b i sung NH4"^, NO2", NO2' vdi ning dd cao cung nhu chftng phit triln USn mdi trudng liSn hop 3 loai NH4^, NO2", N02' ti-ong klii dd ddng HP3.2 ed him lupng tich luj PO4 cao nhit
TT T£n dong vi tdiuan 01 TODl.l 02 TOD2.3 03 TND1.4
Bang 3: Ba dong vi Ichuan co Idia
raMNH4+
900 300 700
mMNOi.
90 100 100 nSng ticli luy polyphosphate cao TT Ten dong vi kliuin Tdng lugng
01 TL2.3 02 HP2.3 03 HP3.2
PO4'' (mg/l) LuQfng PO4 tyt do 11,26
10,56 46,68
mM NO3- mM tSng h^p ba
600 200 900 300 900 400 (mg/l) Lurong PO4 tich luy (mg/l)
0,88 0,55 1,10
10,33 10,01 45,58 Tit ci 6 dong vi khuin deu dugc nhin difn d
bdng 1500 bp Uen phi difn di cfta PCR-16S rRNA dupe nhan len tft DNA cfta chftng (Hinh 2) va dugc xic dinli ddng TODl.l ding hinh vdi JF799886 Enterobacter sp. CIFRl D-TSB-9-ZMA mftc do 99% va ddng TOD2.3 tuong ddng d mftc 99% vdi ddng HQ259961 Klebsiella variicola sUain 7 &
dong EU884439 Enterobacter sp 12, ddng TNDl 4
1000 bp
ddng hinh vdi ddng FJl 89785 Enterobacter sp.
CSB08; trong khi ddng vi khuan ti'ch luy poIy-P TL2.3 tirang ding d mftc 98% vdi dong JX025736 Bacillus cereus strain VPl 1 va hai dong HP2.3 &
HP3.2 ddng hinh d mftc df 99% vdi dong JF505965 Exiguobacterium mexicanum sUain KNUC9031 vi ddng FJ976560 Acinetobacter soli stram LCR52, tiieo tou tu
Hinh 2: Phd difn di eia sin pham PCR duge nhan len tir DNA cua cac dong vi khuin chuyin hoi N (trii) va vi khuin tich Iny poly-P (phii)
Ghi chu: M-thang chudn, I, 2 vd 3 la 3 dong vi khudn TODl.l, TOD2.3 vd TNDL4, 4 din 9 Id 3 ddng vi khudn lich lup poly-P, 4-5: ddng TU. 3. 6- 7: ddng HP2.3 vd 8-9: dong HP3.2 vd 10: ddi chung dm
Tft Hinh 3 cho thiy cay phi hf chia toanh 2 nhinh trong do itoinh 1 bao gdm 2 ddng TNI .4 vi TODl.l gin vdi chi Enterobacter sp. ttong khi nhanh 2 cd ddng TOD2.3 gin vdi chi Klebsiella vi Enterobacter sp.
Chiing vi khuin chuyen hoi nita v i vi khuin tich luy poly-P vio ttong nude ri rac cd ndi^ dp ammonium ban diu la 400 mg/l sau 5 ngdy cho tiiiy hdn hgp 2 ddng TNDl.4 v i TL2.3 vi hdn hpp 2 ddng TND1.4 v i HP3-2 lam gidm lupng
ammonium xudng 5,6 v i 5,1 mg/l, theo thft ty (Hmh 5) trong khi dd bdn hpp 2 ddng TND1.4 v i HP3.2 giam lupng orthophosphate lidn tyc vi dat mftc thip nhat vdo ngay thft 5 (Hinb 6), nhu viy hdn hpp 2 ddng ehuyin hoi dam Enterobacter sp. TNDl.4 vd ddng tich Iu9 polyphosphate Acinetobacter soli HP3.2 hidu qui nhit ttong vifc logi bd ammonium vd orthophosphate uong nude ri rdc sau 5 t^dy chftng vi khuin Uong md hitto thi nghifm IL.
Tgp chi Khoa hgc Trudng Dgi hgc Cdn Tha PhSn A: Khoa hgc Tif nhien. Cdng nghi vd Moi irudng. 33(2014): 117-124
FJ1897B5_Enlerobaflef_sp._CSB08 7N1.4
- EF4191B1_Ertaobadef_sp._fncp11b
— T O D l . l
- JR99886_Ente!Dbadefjp._aFRl_D-TSB-9-ZMA 10013
- f«7512O0„Klet)siellajneunioniaejlrain_RSN19 H02B961_Kldisiela_wriicdajtrain_7 100 L EU88«39_ErtHii&actef_sp._12
Mhanhl
Nhinh 2
Hinh 3: Ciy pha be trinh biy mii quan he giiia cac ddng vi khuin chuyin hoa nito* dya trdn trinh tur 16S rRNA (phmmg phap neighbor-joining) vdi 1000 lip lgi (bootstrap)
Ciy phi hf ttinh biy mii quan hf cua ddng HP2.3 gin vdi ehi Exiguobacterium vi ddng TL2.3
vdi chi Acinetobacter va Bacillus (nhdnh 1) uong khi do ddng HP3.2 d nhinh 2 (Hmh 4).
J^5[l5%5Ji(igjobKter'iJirje):icariyiiijlfaBi_l(MJC9()31 JF505Ei4G.Ei(igii[^3Cleiiiiiii_auraiiliaaim_slHn_i{NUC3[|l2
iFJ3J656[IAciiet[ibactersiilistrahLCR52 llHlEFt7g435_iM)l)ai:tef_lHylyi_sliaii_3R22 - JMM3>cisjisetitaycoiilesjtraiii_)[ZP6S<
,m
KI .WBSTaii.Bacius.cefais.sftain.VPII HP3.2
Hinh 4: Cay pha he trinh bay mii 35 quan hf giiia cac dong vi khuin tich Iny poly-P dua trdn trinh ti^l6S rRNA (phuang phip neighbor-joining) vdi 1000 Igp Igi (bootstrap) Ngoii ra khi syt khi lidn tuc lam cho lupng khi
ammoniac (NH3) tooat ra vi bay vio khdng khi vi thi lupng khi NH3 d n^ifm toftc 2 (chi sye khi) cao nhat trong tftng ngay. Trai lgi nirdc ri rac khdng sye kin' (NTl) eo lupng khi ammoniac tooat ra it nliat, dieu ddc biet nude ri rac ed chung vi khuin chuyin hoi nito lam cbo kbi NH3 tooit ra nliung it ban nghif m thfte 2 (sue kbi") vi luong nito eon lgi dupe vl khuan sft dyng nhu nguin dudng
chat thay vi thoat ra ngoai lam 6 nhiem khdng khi.
Hon hpp 2 dong ddng chuyin hoa dgm Enterobacter sp. TNDl.4 vd dong tich luj polypho.sphate Acinetobacter soli HP3.2 cd lupng khi ammoniac toil ra it nhit (Hinh 7). Die bidt 2 dong TODl.l va ddng HP3.2 su dung nhilu P dd tan nen khi bd sung vio thi nghifm (Hmh 6) lam cho lupng P d ngdy 0 toip hon cie n^iem toftc khae.
Tap chi Khoa hoc Trudng Dgi hoc Cdn Tha PhSn A: Khoa hgc Tu nhien. Cdng nghi -vd Mdi trudng: 33 (2014): 117-124
chwisvi Muan
Hinh 5: Hifu qua ciia vi khuin chuyin hoa nitu va vi khuin tich liiy poly-P trin ham lupng ammonium (mg/l) trong nude ri rac theo thdi gian
ngay isu km clii!r^ vl khuAn
Hinh 6: Hifu qua ciia vi khuan chuyen hda nitava vi khuan tich luy poly-P trin ham lupng orthophosphate (mg/l) trong nude ri rac theo tbdi gian
Ghi chu- NTl: ddi chung (khong sue khi), NT2: ddi chung (s\ic khi), NT3: dong TODl. 1 vd ddng TL2.3. NT4: ddng TODl.l vdddngHP3.2. NTS: ddng TND1.4 vd dong TLZ3, NT6: ddng TND1.4 vd ddngHP3.2 [NTS. NT4. NTS, NT6 deu s\ic khi liin tiic]
I'gp chi Khoa hgc Trudng Dgi hgc Cdn Tho Phan A: Khoa hgc Tu nhien, Cdng nghi vd Mdi trudng: 33 (2014): 117-124
400 - 350 - 300
200 150 -
5 0 •
U -
. - ^ - - 1 » -2 — A - 3
^*^
^^
r-- /J^^ /
j ^ ^ . . .
-;-^r^;3*!*^
0 1 2 2 4 5 ngay s a u hhi chung vl khuan
Hinh 7: Hifn qua cua vi khuin chuyen boa nitorvi vi khuan tich luy poly-P tren ham lupng khi NH3 (mg/l) thoat ra tir nude ri rac theo thdi gian
Ghi chu: NTl: ddi chung (khong sue khi). NT2- ddi chung (sue khi). NT3: ddng TODl. 1 vd ddng TL2.3.NT4: dong TODl. I vd ddng HP3.2. NTS: ddng TNDL4 vd ddng TL2 3. NT6: ddng TNDl 4 vd dong HP3.2 [NT3, NT4, NT5 NT6 diu sye khi liin luc]
Chung vi khuin chuydn hoi nito lim giam ham lupng Nita ting (TN) so vdi dii chftng, sue khi (NT2) gidm dugc 75,31% ttong khi cac ddng vi khuin ehuyin boa nita tix 79,01% din 90,12%
ttong khi do sue khi Igi lim ting him lupng P tdng
Bang 4: Hieu qui ddng vi khuin chuyen hoa nitu va vi kbuan tich luy poly-P trin ham lupng TN, TP, Nitrite va Nitrate (mg/l)*
(TP) va hdn hpp cac dong TODl.l+ddng HP3.2, TND1.4+d6ng TL2.3, ddng TND1.4+d6ng HP3.2 giam lugng TP Udi lai him lupng niUite va nitrate Uong nude ri rac qua toip (Bdng 4).
Chi tilu NTl NT2 ., ^,\]\ NT3 ., ^,\]\
giam(%) giam(%)
Ti If giam Ti If giam Ti If giam
(%) (%) (%)
"" " 46,7 87,6 37,4 TN 378 93 75,3 79,4
TP 16,5 3J,4 - ^ - 7 7 ^ NiUite 0,05 0~ - - 1 0 0 "6;0I Niuate 1,05 0,17 83,3 0,17
79,0 80,0 83.8
42,0 8,0 0,03 0,14
88,9 51,4 9,8 40,0 0,02 86,7 0,14
40,4 12,1 60,0 0,07 86,7 0,10
90,1 26,5 90,0
* Phan lich Igi Trung tdm Ky thudt vd Ung dung Cdng n Ghi chu: NTl- ddi chung (khongsue khi), NT2- ddi chung (i
TtpDl 1 vd ddng HP3.2. NTS: ddng TNDL4 vd ddng TL2.3. NT6: ddng TNDL4 vd dong HP3.2 [NT3. NT4, NT5 NT6 diu sue khi liin tficj
}, Sd Khoa hgc va Cong nghe TP. Cdn Tho g (sue khi), NT3: ddng TODl. 1 va ddng TL2 3, NT4: ddng
Nhin chung hdn hpp 2 ddng TND1.4+HP3.2 hifu qud nhit Uong vifc logi bo ammonium, nittite, nitrate, ortoophospbate vi TP kl din 2 ddng TND1.4+ddngTL2.3.
Xft ly nita vi photpho trong nude ri rac di dugc Le Vin Cat (2007) dl nghi tft liu bing phuong phap li va hoi hpc d Vift Nam tuy chua dat kit qua ni^ man nhung tic gia cung da dl cap din mdi nguy hiem tft nude ri ric giiu nita vi photpho; Su et al. (2001) la nhftng nhi khoa hpc diu tien nhin
difn v i ftng dung vi khuin khft dam Pseudomonas stutzeri de logi bd amoni ttong nude thdi ttgi heo d Dii Loan ttong dieu kifn cd oxi (sye khQ, chiing tdi ciing da dl nghj logi bd nita ttong nude ri rac giiu hftu CO bing vi Idiuin khft dam vi khuin Pseudomonas stutzeri vi Acinetobacter Iwoffii (Cao Ngpc Difp va Nguyen Thi Hoing Nam), gdn diy Cao Ngpc Difp et al. (2013) da ftng dyng hai dong vi khuin chuyen hod nita di dumig Pseudomonas smtzeri D3b vi dong vl khuin tich luy poIy-P Bacillus subtilis DTTOOIL loai bd N v i
Tgp chi Khoa hgc Trudng Dgi hgc Cdn Tha Phdn A: Khoa hgc Tu nhien, Cdng nghi vd Mdi tnrdng. 33 (2014): l!7-lU P Uong nude ri rac rit hifu qui, ket qui cfta chung
tdi trudc diy (Bui Thi Vinh et al. 2011) cung cho tody dong vi khuin chuydn hoi nita Enterobacter sp. PI I loai bd ammonium ttong nude thii nhi may chl biln sfta rit tdt.
4 KET LUAN
- Trong nude ri rdc chfta nhieu ddng vi khuin chuyin hod nito va vi khudn tich luy polyphosphate trong dd cd nhidu dong vi kliuan co do hftu hidu cao.
Kit hop hai ddng vi khuin chuyin hoa nita Enterobacter sp. TND1.4 vi dong vi khuin tich luy polyphosphate Acinetobacter soli HP3.2 logi bd N va P ttong nude ri rdc trong dieu kien sye klii d md hinh thi nghifm phdng toi nghifm dat tieu chuan Bl vl ammonium, Tdng N, nittite, nitrate, orthophosphate, Ting P eua QCVN 24 vd QCVN 25: 2009/BTNMT.
L d i CAM T ^
De tii duge thyc hien vdi sy hi Up kinh phi cfta Sd Khoa hpc vd Cdng nghe Thinh phd Can Tho.
TAI LI?U THAM KHAO
1. Bui toi Vinh, Hi Thanh Toan va Cao Ngpc Difp. 2011. Phdn lip va nhin difn vi khuin chuyen hda nito tft chit thdi trai nudi bo sfta, chit thai sfta va ung dyng trong xft ly nude thdi nhi mdy sin xuit sfta. Tgp chi Khoa hpc-Dgi hpc Cdn Tho 18a: 194 - 200.
2. Cao N^pc Difp vd Nguyen Thi Hoing Nam.
2012. Ung dung vi khuin Pseudomonas stutzeri vd Acinetobacter Iwoffii logi bd amoni trong nude thii tu rdc hftu ca. Tgp chi Khoa hpc- Dgi hpc Cin Tha 22b: 1-8.
Cao Ngpc Difp, Nguydn Thj Kim Em vi Nguyin Trin Hing Phuc. 2013. Kha ning loai bd nita vi photpho Uong nude ri rdc cfta vi khuin khft dam di dudng v i vi khuin tich luj polyphotphat. Tap chi Ndng nghiep v i Phdt ttiln Ndng todn, 2: 16-24.
4. Eixler S., U. Selig and U. Karsten. 2005.
ExUaction and detection methods for polyphosphate stton^e in autotrophic planlaonic oi^anisms. Hydrobiologia, 533(1): 135-143.
5. Le Van Cit. 2007. Xft ly nude toai giiu nito v i phospho. Vifn Khoa hpc vi Cdng nghf Vift Nam. Tamura, K., D. Peterson, N.
Peterson, G. Stecher, M. Nei, anA S. Kumar.
2011.MEGA5:
6. Tamura, K., D . Peterson, N . Peterson, G. Stecher, M. Nei, and S. Kumar.
2011. MEGA5: Moleeuleu Evolutionary Genetics Analysis using Maximum Likehood, Evolutionary Distance and Maximum Parsimony Metoods. Moi. Biol.
Evol, 28, pp. 2731-2739.
7. Wang, D.B, X.M. Li, Q. Yang, G.M. Zeng, D.X. Liao and J. Zhang. 2008. Biological phosphoms removal in sequencing batch reactor with single-stage oxic process.
Bioresource Technology, 99(13), pp. 5466- 5473.
8. Zhao, B, Y.L.He, J.Hughes and X.F.Zhang.
2010. HeteroUophic nittogen removal by a newly isolated Acinetobacter calcoaceticus HNR. Biores. Technol. 101, pp. 5194-5200.
