KHOA HQC CONG NGHE Tfiil u u HOA KHA IMAIXIG TOMG HOP CHAT K £ T TU SIIMH HOC CIJA CHLIIMG VI KHUAIM Bacillus subtUis PO OB B VA THU IMGHIEM XU LY IMUQC THAI TU CQ 5 Q SAIM XUAT BUIM It Tlii Loan^ Cao[.]
Trang 1Tfiil u u HOA KHA IMAIXIG TOMG HOP CHAT K £ T TU
SIIMH HOC CIJA CHLIIMG VI KHUAIM Bacillus subtUis
P O O B B VA THU IMGHIEM X U LY IMUQC THAI
TU CQ 5 Q SAIM XUAT BUIM
It Tlii Loan^ Cao Nggc Di^p^
T6MTAT
Nuoc thai tir co sd san xuat bun vin con chiia mot lugng ldn chat hiru ca nhat la lugng tinh bgt trong gao cdn l^i se gay 6 nhiSm nude nen can phai duoc xir ly trudc khi thai ra moi trudng Chat k6t tu sinh hpc (bioflocculants) la mot hgp chat cao phan tu dugc tong hop trong qua trinh phat tri^n cua cac vi sinh vaL Chiing CO tac dung nhanh chong va an loan cho con ngudi va moi trudng nen dupe nghi&n ciru va img dung
d^ xir ly mrdc thai Chiing vi khuan Bacillus subtilis PO.03.B dugc phan lap lu miu nude thai co sd san xuat hii ti^u My Tho d tinh T\kn Giang, c6 kha nang tong hop chat k^t tu sinh hpc vdi thinh phan m6i truong tdi
uu cho kha nSng tdng hgp chat ket tu sinh hoc gdm Unh bot (1,0%) ure (0,05%) FeCI^ (0.75%) d pH 9 cho ty 1§ k^t tu 79.57% vdi dung dich kaolin sau 5 phut d^ ling, bo sung dung dich CaCl;, va 0.05% dich nuoi sinh
khdi VI khuan Ung dung chung vi khuan nay trong xir ly nuoc thai ca sd san xuat biin da lim giam cac chi
sd BOD5, chi sd TSS, chi sd nito tdng, chi sd photpho tong va ham luong amonium I^n lugt la 17,76% 11.26% 21,87%, 21,67% va 36,61% so v6i chi sd ban dau Chi tieu tdng chit ran la lirng va ham lupng amoni dgt tieu chuan A cita quy chuan QCVN_ 40/2011/BTNMT
TLT khda: Bacillus subtilis, nguon carbon, nguon nitrogen, kd't tu sinh boc, nude tiiai ca sd san xuat biin, pH
I.GNlTHiU
Sg k^t tu trong nude dugc chia lam 3 nhdm
chinh gom: Ket tu vd ca nhu phen chua (aluminium
sulphate va cloride aluminium), ket tu hiru co tdng
hop (cic din xuat tir polyaciylamide vdi
polyethylene Imme vi cic chat ket tu tu nhien nhu
chitosan, sodium algmate) vi ket tu bdi vi sinh vat
(microbial flocculants) Cic chat ket tu hda hpc hiy
CO gii thinh thip nhung lai anh hudng ddn sire khde
con ngudi vi mdi hirdng, chdng han nhu cac hgp
chat k^t hi cua nhom la nguyen nhin gay ra benh
Alzheimer (Kurane etal., 1994), cdn cac din xuat tir
polyacrylamide la dpc td cho he than kinh va li chat
gay ung thu, khd phin hiiy trong lir nhien (Yokoi et
al., 1996) Trai lai, chat ket hi sinh hoc
(bioflocculants) li chat dugc tdng hpp tir vi smh vit,
cd tac ditng nhanh chdng v i an toin cho con ngudi
va mdi truang (Sheng etal, 2006)
Cdng nghe ket hi (Flocculahon technology) da
duoc su dimg rpng rii h'ong linh vuc xti ly nuoc thai
' N'CS chuyen nganh Vi sinh vat hpc, Tnrdng Dai hpc Can
Tho
Vi?n Nghien ciiu vii Phat tri^n Cdng nghe Sinh hoc
Tnidng Dai hpc C5n Tho
dac biet li trong cdng doan tien xir ly ciia nhi^u h$ thdng xu ly nhd uu diem li diu tu ca sd ha tang nhd
vi thdi gian xit ly ngin Su ket tu sinh hpc li su ket
tu cic vit chat la limg trong nude giup lam giam cic chi tieu nhu COD TSS vi tir do giup lim giam mot phan dp diic cua nude thai trudc khi dupe xu ly
bing phuong phip khic (Gong et al, 2008) Mpt
sd nghien cun cho thiy img dung chat k^t hi smh hpc trong xu ly nude thai nhir chat ket tu sinh hpc
cd ngudn gdc tir vi sinh vat dupe sir dung d^ xir ly
nude thil tir xi nghiep nhupm (Zang et al., 2002; Deng et al., 2005) cac chat la limg vd co (bentonite, dat set, Ca(OH);,, aluminum oxide), (Shih et al., 2001; Yun etal., 2007), acid humic (Zoubouhs et al., 2004) vi cic chat la lung khic (Salehizadeh et al.,
2000)
Trong nghien cuu nay, da xie dinh didu ki^n tdi
uu hda cac y&u td inh hudng den su tdng hgp chat ket til sinh hpc cung nhu ty le k^t tij sinh hoc cua
chung vi khuin Bacillus subtilis PO.03.B duoc phin
lap tir nude thai cua cac ca so sin xuit hu tieu My
Tho, tinh Tien Giang (Le Thi Loan et al.„ 2018) va
thtr nghiem hieu suit xti ly nude thai cua cic co sd san xuat bun thdng qua xie dinh cac chi deu B 0D-,
vi TSS, him luong nita tdng, ham lugng photpho tdng (P) va ham lupng amonium trong thanh phan
63
Trang 2nuoe thai
Z VAT LfilVAPWDNG PHAP
2.1 Phuong t i ^
Su dung chiing vi khuan BaciUus sutibs PO.03.B
phin lip tu nuoc thai ciia cac ca so sin xuat hu tieu
My Tho tinh Tien Giang (Le Thi Loan etal 2018)
thoi gian nudi sinh khoi vi khuan sau 96 gia cho ly le
kdt tu cao nhal cd d mdt -^c vi khuan tren 5 x U'
tuong tmg Ol),,., = 1.24 ± 0.(12,
Nude thai cua U bun D U ^ T \ so 32/10 duong
Bui Him N'ghia K\7 phudng Binh Thuy Q Binh
Thuy TP Cin Tho) Ket qua ham lupng cac chi Ueu
ban dau cua ngudn nude thai (sau biogas) duge Uinh
bay o b i n g 1
Bing 1 Him Itrgng ckc chi tifiu ban diu cua nude
thai ca sd sin xuit biin .,
Chi lieu
pH
Aiiionium (N_NH,*) mg/L
Phosphate (P_PO,^) mg/L
Djin tdng (TKN) mg/L
Phospho long (TP) mg/L
Nhu cau oxi smh hoc (BODO mg 0^/L
Tong chat ran la lung (TSS) mg/L
Ham
lumlg
4 0 5 '
1 1 2 ' 179,2 = 17.7 -•
1520 = 36,4
-(Nguon: ' Phan tich tai ITN I / sinh vat mdi
tnrdng - 1 'icn SC&PT Cdng nghd Smh hoc, Trudng
D^i hpc Cin Tlia - Phan ti'ch tai Phdng tiii nghidm
chuydn sau - Trmmg Dai hoc Can Tha)
22 Xie dinh ty 1$ k^t hi sinh hpc
Chiing vi khuan Bacillus subtilis PO.03.B diroc
nudi cay trong 50 ml mdi truong tdng hop chat kdt tu
sinh hgc polysaccharide (Deng etal 2003) vdi thinh
phin cac chat gom 10 g glucose 5 g KH^POj 2 g
K HPO, 0,1 g MgSO,.7H.O 0,1 g NaCl, 0.5 g
carbamide, 0.5 g yeast extract, 20 g agar (bo sung
khi dd mdi truang thach), nuoc cat bd sung du 1 lit
va dieu chinh gii tn pH=7 Irong binh tam giac 100
ml Lie ti-en miy lie xoay vong d tdc do 1(>0
vdng/phut d nhiet dn 30 C
Dung dich vi khuan sau thdi gian 4 ngay nudi u
dupe sir dung de kiem tra kha nang ket tu bang hdn
hop gdm 90 ml dung dich kaohn (5 g/l) 10 ml dung
dich CaCl 1% va bd sung 100 pi dung dich vi khuan
vdi mat -^o >10' td bao/nil (ti le 0.1%) Hdn hop duac
khuiy deu 60 vong/phut trong 30 giiy bang miy
Rhu.y Hoa Ky sau do Ai ySn = Ph" n " ^ ^^ ^ ^ ^ ^ trtn each mat ntnx- 2 cm iunc hu do - ^ * ^ * *
due quang ph6 W (Spectrophotometer) " ' ' " ^ ^ ^ ^
550 « , , (Deng etal 20O3) Mau f ' ^ ' ^ f ^
tu«T,g ttr nhimg khong bo sung dich v khuan, ty le ket til diroc tinh theo cong thirc:
ODa„d,.^ OD,j„
X 100%
ODa.,d„«K-Phirong phap nav duoc sir dung de xac dinh ty 16
ket tu smh hpc cua chung vi khuin Bacillus subtilis
PO.03.B trong tat ca cac thi nghiSm
2.3 Tdi mi hda cac yiu td anh huong d & k h i nang tdng hpp chit kit tu sinh hpc v4 hi^u suit Ut
tu ciia chiing vi khuin Badllus subtilis PO.03.B
Cac thi nghiem dupe bd tri theo the thiic ngiu nhien hoan loan de khao sat anh hirong cua pH moi tnrong, thanh phan moi tnrong nudi u, cac mudi kim loai va nong dp dung dich vi khuan bo sung d & hieu
suat ket tu sinh hpc cua chiing vi khuin Bacillus subtilis PO.03.B Tir dp xac dmh dirpc cac di^u kien
thich hpp de chiing vi khuin cho hieu qua kA tu sinh hpc cap nhat
2.3.1 Khad sat anh hudng ciia pH mdi tiirdng nudi u
Anh hirong ciia pH mdi tnrdng nudi u d^n kha nang tong hpp chit ket tu sinh hoc ciia chiing vi
khuin Bacillus subtilis PO.03.B dupe khao sat trong
khoang pH tir 4 den 11
Thi nghiem duoc thuc hien gom 9 nghiem thiic tirong img voi mdi nghiem thlic la mpt gia tri pH khac nhau: 4, 5, 6 7, 8, 9 10, 11 M6i nghiem thirc dugc thuc hien vdi 3 lin lap lai Chiing vi khuin dupe nudi trong moi tnrcmg tong hop chit ket tu sinh hpc
polysaccharide (Deng etal, 2003) vdi gia tri pH dupe
dieu chinh phii hpp d timg nghiem thlic bing dung dich HCl IM hay NaOH IM (Merck) Lie tren may
lac xoay vdng p tdc dp 160 vdng/phiit a nhiet dp
30"C Sau thdi gian 1 ngay, dung dich vi khuan se duoc danh gia kha nang ket tu theo phirong phap da dupe md ta d muc 2.2 Tu dd chon diroc gia tri pH mdi truong nuoi smh khdi thich hpp chp su tong hop chat ket tu sinh hoc dat hieu qua kit tu cao nhat
2.32 Kliao sal anh hudng aia ngudn carbdn, nitrogen va khoang vd co
Cac nguim carbon, nitrogen va khoang vd cp
N O N G N G H I E P V A PHATTRIEN N O N G THON - KY 1 - THANG 10/2019
Trang 3khao sat dupe trinh bay frong bang 2 Cic ngudn
dmh dudng nay duge lua chon vi ngudn X hiiu ca
ph6 bi^n la Yeast Extract (Merck) vi cac hda chat
con 1^ cua Trung Qudc
Phuong phip tidn hinh bang each nhin sinh
khdi chiing vi khuan trong 20 ml mdi tnidng vdi J
dinh hieu qua ket tu vdi dung dich kaolin theo nhu
md t i d muc 2.2 Phdi hgp ba ngudn dmh dudng (3 ngudn carbon + 4 ngudn nitrogen + 1 ngudn khoang
vd ca) cd 48 ngluem thuc khic nhau va mdi nghidm thlic dirge thtrc hien 3 lin lap lai (Bing 2) Ket qua cudi cung chgn ra dirge mdi tnrdng toi phdi hgp cua ba ngudn carbon, nitrogen vi khoing uu cd ngudn carbon, nitrogen vi khoang vd ca th, v6 CO dtrdn, vdi gia tri pH da chgn, nhiet dd 30"C, lie hgp cho vi khuan tdng hgp chit ket tii sinh hoc cho
160 vong/phut, sau thdi gian 1 ngay tidn hanh xie ty le ket hi cao nhat
Bing 2 Cic ngudn carbon, nitrogen v i khoang vd ca dugc bd sung trong mdi tnrdng nudi sinh khdi chiing vi
khuan BacillussubtiUsPO.033
Ngudn carbpn
Glucpse (1%)
Sucrose (1%)
Tinh bot (1%)
Ngudn niU'Ogen Acid glutamic (5%) Yeast extract (0.05%) Urea (0,05%) (NH.,).^0, (0.05%)
Nguon khoang vd co KCl (0,5%) FeClj (0.5%) CaCl, (0,5%) K.,HPO,(0,2%) + KH.,PO,(0,5%)
2.3.3 Khio sit anh hudng cua ty le nguon
carbon, nitiogen vi khoing vd ca ddn hidu qui tong
hop chat kd't tu sinh hoc
Bing 3 Nghidm thtic b6 tri theo ki^u hoin toin ngiu
nhl6n khao sit inh hudng cua cic t^ Id carbon,
nih-ogen vi khoing v6 ca d^n hi^u qua k^t hJ smh
hgc
Nguon
carbon
C (a,%)
C (aj«)
C (aj*)
Ng
N(b,%)
1
2
3
10
11
12
19
20
21
udn nitrogen
N(b2%)
4
5
6
13
14
15
22
23
24
N (b;,%)
7
8
9
16
17
18
25
26
27
Ngudn khoang vd
C O
M (c,%)
M (c.,%)
M (c.,%)
M (c,%)
M (c,%)
M (c-,%)
M (c,%)
M (c.,%)
M (Cj%)
Ghi chii: C: nguon carbon: a: muc dd (%): N:
ngudn nitiogen: b: muc do (96): M: ngudn klioing vd
ca.amucdp (%)
Nhan sinh khdi chung vi khuin frong mdi
tnrdng gdm 3 nguon carbon, nitrogen va khoang da
dugc chgn b thi nghiem fren Moi ngudn dinh dudng
se duoc chia thanh 3 miic dp khac nhau va phdi hgp
tao ra 27 nghiem thtrc vdi ty le carbon, nitrogen va
khoang vd cabo sung khac nhau (Bang 3)
Chiing vi khuan duge nhin sinh khdi U'ong dng
fancol 50 ml chua 20 ml mdi trudng td hgp tii 3
nguon dinh dirdng vdi ty 1^ khic nhau, di^u chinh vd gia tii pH cho ty le ket tu cao nhat da chgn, u lie 160 vdng/phut d nhiet dd 30"C Danh gii kha nang kdt tu vdi dung dich kaolin, phin ti'ch hoi quy tuyen tinh 3 nhin td tir do chon ra nghidm thiic cd ty 1§ bd sung thich hgp cho vi khuan tdng hgp chat ket tu sinh hgc cho ty le ket tu cao nhit
2.3.4 Khio sat inh hirdng ciia ndng dp djch vi khudn bo sung
Tien hinh kiem fra khi ning k^t tu sinh hgc cua
chung vi khuan vdi kaolin b nhtmg nong dp khac
nhau: 0.002; 0,005; 0,01; 0.05; 0.10; 0,15 vi 0,2% Dinh gii khi nang ket tu bing cich bd sung 10 ml dung dich 0,75% mudi kim loai da xac dinh d thi nghidm fren vao 90 ml dung dich kaolin (5 g/l) vao cdc 250
ml Sau dd them dung dich vi khuan tdng hpp chat kdt til sinh hpc d nhiing nong do khic nhau Mau ddi chung thuc hien tuong tu nhung khdng chung vi khuan tong hgp chat ket tu sinh hgc Khuay d^u hdn hop fren frong 30 giay bang may khuiy tir, giu yen hdn hpp frong 5 phut, lay phan trong each mat nude
2 den 3 cm xac dinh chi sd OD d bude sdng 550 nm Tinh kha nang kdt tu va chgn ndng dd vi khuin tdng hgp chat ket tii sinh hpc cd kha nang ket tu cao nhat ung vdi timg ddng vi khuan ket tu sinh hgc Thi nghiem gdm 7 nghiem thuc vi mdi nghiem thuc duoe thuc hien vdi 3 lin lap lai
2.3.5 Thti nghiem hidu suit xuly nudv Uiai ca
sa sin xuat biin ciia chiing vi khuan Bacillus subtilis PO.03.B
Ap dung cic didu kien thich hgp ve do pH, mudi
65
Trang 4kim loai bd sung, ngudn dinh dudng nudi va Ueu
luong dich VI khuan da tim dirge cho chung \i khuan
Bacillus subtihs PO.03.B d^ xu ly nude thai ca sd san
xual bun a quy md phdng Ihi nghiem (bmh 10 In)
Qui Uinh nhu sau: S lit nuoc thii ca so sin xuat bun
dupe chua frong binh cd the tich 10 Ut chua, bd sung
mudi khoang kim Ioai, dung dich \i khuan thich hop;
syc khi trong 5 phut (cac thanh phan frong nude thai
va mudi kim loai vi dich vi khuan trgn deu nhau de
phan ung xay ra) d^ yen 60 phuL Thu phin nude
trong phia frdn de xac dinh cic chi tiSu BOD5 TSS,
him lugng nito long, him Iuong photpho tdng (P) va
him lupng amonium ciia nude thai sau xu ly b Phong
thi nghiC'm chuyi'ii sau - Dai hoc Can Tha
2 '{.t} Ghi nhin kdt qui va vuly
Sd li^u thi nghidm dugc phin lich bing phan
m^m thdng ke Statgraphics Centurion XVI.I va
Microsoft Office Excel 2013
8.K£TQUAVATHAOLIUN
3.1 K^t qui tdi uu hda kha ning tdng hgp ch^t
k^t ti4 smh hgc cua chung vi khudn Bacillus subtihs
PO.03.B
S.Ll.GiatiipH
19,10 i
ISSb
CV^3,32?i
Hinh 1 Anh hirdng cua pH mdi tnidng nuoi smh
khdi d&i khi nang tdng hgp chit k^t tu smh hgc
chung vi khuin Bacillus subtihs PO03.B
Ghi chu: Shimg so tidn mdi cdt theo sau ciing 1
chir khong kbac } nghi.i thdngkd dmuc dd 1%
Nhan td pH cua mdi truong nudi cay dupe cho la
nhin id xac djnh di^n Uch trao ddi cua cic te bao vdi
nhau ciing nhu ii cic cau ndi cdng hda tii - yeu in
anh hudng den su hlip thu chat ^ - h f " * ^ ^ * ^
phan m,g cua enzyme (Xui etal ^ « « ; ' " ' " 7 ™ ^
nudi sinh khd, p cac gia tn pH khae "hau * ' anh huting khac nhau den kha nSng tdng hPP c >at ket tu smh hpe (Salehizadeh va ShojaosadaU JOlU)
Tv \e kft tu kaolin cua chiing vi kltujn Bacillus subtihs PO.03.B la 19.10% d pH 9 sau khi thay dpi pH
mdi hmmg nudi eiy d cac gia tri pH khac nhau thi ty
le kdt tu cung thay ddi khac nhau (Hinh 1) Khi dltpc nudi cay hong mdi trudng cd gia tri pH tir 5 d t a 8 thi chiing VI khuan chp ty le kdt tu cap va dat ty 16 kft tu cao nhat p gia hi pH 9 (19.10%) khac biet cd y nghia
sp vdi p cac gia tri pH khac Mdi chung vi khuan thich hop vdi khoang pH nhat dinh va dat ty Id kft tu cap nhat tai mdt gia tn duy nhat Theo kft qua thi nghiem, pH 9 duoc chpn la pH mdi htrdng nudi cay ddng vi khuan de thuc hidn cac thi nghiim tiSp thep
3.1.2 Ngudn carbon, nguon mtivgen vi khoang vdca
Cac ngudn dinh dudng khac nhau trong thanh phan mdi htrdng nudi sinh khdi cd anh hudng khac nhau den kha nang tdng hpp chat kft tu sinh hpc eiia
chung vi Idiuan Bacillus subtilis PO.03.B
Kft qua khac sat sir anh hudng cua cac ngudn carbon, nitrogen va khoang vd co den ty le kft tu cua
chung VI khuan Bacillus subtihs PO.03.B (Bang 4)
cho thay ty le kft tu cap nhat dat dupe la 78,21% vdi ngudn carbon la tinh bdt (1%), ngudn nitrogen la YE (0,05%), ngudn khoang vd co la KCl (0,5%) Ngoai ra, thanh phan mdi truong gdm tinh bdt 1%, ure (0,05%), KCl (0,5%) cung cho ty le ket tu cao la 77,27% v4 khdng khac biet thdng ke vdi nghiem thtic cd ti 16 kft tu cao nhat nhung chi khac biet nguon N la ure
va YE, chpn ure vi gia thanh se re hon so vdi YE
Theo Zheng etal (2008) ddi vdi Vl khuan BacillusSf
F19 thi ngudn carbon thich hop la sucrose (8g/l) va ngudn nitrogen la dich trich nam men (0,25g/l) Vi
khuin Bacillus subtihs MSBN17 chp ty 16 kft tu cao
nhat la 92,07% khi dupe nuoi trong mdi hudng co nguon carbon la dudng thdt ndt, 89,04% vol ngudn nitrogen la NH^NO, va 88,36% vdi ngudn khoang v6
cp la NaCl (Sathiyanarayanan et al L'Ol'',) Qua kft
qua nghien eiiu nay chp thay chung vi khuan cd the thich ung vdi nhieu nguon carbpn, ngudn nitrogen va nguon khoang vo co khac nhau Tuy nhiSn chi cd mot ngudn carbon, nitrogen va khoang vd ca thich hop nhat eli„ su tdng hpp chat ket tu smh hoc cho tv 1' ke, u, ,.a„ „ha, V, vay chpn nK„„n j , , , , , j ^ , ^ g
Trang 5giro linh bpt (1%) ure (0.05%) FeCl, (0,5%) lam mdi cac thi nghiem tiep theo
tntdng nudi sinh khdi chiing vi khuan de thirc hien
Bing 4 Anh hudng dia ngudn (%i) carbon, (%) nitrogen va (5S) khpang vd ca d & ty 1$ kft tij (%) cua chung v
khufa BadBus sublih'sPO.03.B
Kyhidu
Bacillus
subtilus
PO.03.B
LSD„
(D)-0,92
LSD„
(N)=l,23
CV% = 3,23
Ngudn
carbpn
Glucose
(1%)
16,86%
Sucrpse
(1%)
23,15%
Tinh bdt
(1%)
57,27%
Ngudn nitrogen Glutamate
(5%) 16,17%
11,26%
12,37%
13,49%
27,71%
13.37%
05,98%
19.08%
72.87%
55.18%
64,65%
67,77%
Ure (0,05%) 11,41%
36,71%
12,45%
07,15%
14,95%
15,49%
54,14%
03,66%
77,27%
71,77%
73,45%
72,85%
(NH,).50j (0,05%) 10.19%
08,23%
10.27%
19.89%
09,56%
64.73%
15.41%
17.09%
71.27%
73,07%
17,65%
18,05%
Yeast ex
(0.05%) 19.81%
08.41%
46.01%
24.89%
10.29%
21.995 64,49%
12.47%
78,21%
07.37%
58,79%
36.22%
Ngudn khoang
vd CO
KCl (0.5%) CaCl,(0,5%) FeCl, (0.5%) K,HPO,(0,2%) + KH.,POj(0,5%) KCl (0.5%) CaCl2(0.5%) FeCl.i (0,5%) K,HPO,(0,2%) + KH,PO,(0,5%) KCl (0,5%) CaCl,(0,5%) FeClj (0,5%) K2HPO,,(0,2%) + KH2PO,(0,5%)
3.1.3 Ty Id tinh bdt, ure va FeClj
K^t qua tdi im hda ty le tmh bdt, ure vi FeCl^ bd
sung frong thanh phan mdi tiirdng nhin sinh khdi
chiing vi khuan Bacillus subtilis PO.03.B dugc tiinh
bay d bang 5 cho tliay d cic nghiem thirc gom tinh
bot (1%), urd/YE (0,05%) va KCl (0,5%) khic biet cd y
nghia so vdi cic nghiem thiic cdn Iai tuy nhien
nghidm thuc tinh hot (1%), ure (0,05%) va FeCl,i
(0,596) thap hon hai nghidm thiic fren nhung khdng
nhidu va xet ve mat kmh te thi su dung FeClg (0.5%)
se re hon la sir dung KCl (0,5%) b sd luong ldn Dieu
nay chung to thanh phin dinh dudng frong nude thii
hu h^u cd nhidu tinh bdt vi su bo sung ure trong qua
trinh che bidn hu tidu lim ting kha nang tdng hgp
chit k^t hi smh hgc tii vi khuan ddng PO.03.B Tir
ket qua thi nghiem chpn nghiem thiic tinh bdt (1%),
ur^ (0,0596) va FeC\^ (0,05%) la thinh phin mdi
tnrong nudi cay chiing vi khuan Bacillus subtilis
PO.03.B CO ty le ket tu cao nhat
Sir dung cac sd heu ket qua thi nghiem fren
phin tich tim ra phuong tiinh hdi qui nhi^u bidn
bang chuong trinh Statgraphics XVI, dua fren
phuong trinh xac dinh duoe nghiem thirc tdi uu nhit
cho mdi tnrdng nudi sinh khdi chung vi khuan
Bacillus subtilis PO.03.B K^t qui phin tich tuong
quan cho thiy ty le ket tu (%) cd tuong quan vdi 10 bidn theo phuong frinh:
Ti le ket tij (%) = 117.012 • 100,541*Ham luong FeCU - 41.4311*Ham luong Tinh bot - 101,32*Ham Iuong Ure + 13.264*Ham Iuong FeCl2*Ham luong FeCU + 119,49*Ham Iuong FeCl2*Ham luong Tinh bot - 22,5347*Ham luong Tinh bol*Ham Iuong Tinh bot + 137.374'Ham luong FeCl/Ham luong Ure + 138,394*Ham Iuong Tinh bot*Ham Iuong Ure -5.26692*Ham luong Ure*Ham Iuong Ure 180,288*Ham Iuong FeCI/Ham Iuong Tinh bot*Ham Iuong Ure
Dira vao bang phin tich ANOVA, co y nghia
thdng ke giira cac bien a miic dp tin cay 99,0%, sd heu
thdng ke binh phuong cua R (R-squared li he sd tirong quan chi ra su tac dgng cua cac yeu td dinh dudng ngudn cacbon, ngudn nitrogen va khoang vd ca) tic ddng den kha nang tdng hgp chit k^t tu sinh hoc vdi ty le kdt tu (%) cua chung vi khuan la 79,52%
Sd heu thdng ke R-squared (adjusted f.f.) phu hgp hon de so sinh cac md hinh vdi cac sd khac nhau ciia
67
Trang 6cac bidn dpc lip la 43.35* Sai so chuan ciJa cac uoc tri nay cd th^ dupc sir dung de x.n dung gi™ ^*" V tinh chp thay ddlfch chuan cua cac sddu la 5.15 Gia bao chp cac quan sat
Bang 5 Anh hudng oia su thay ddi ty If (5(5 tinh Wt, (X) ur«, (%) v4 KCl (%) d & ty 1$ kft tu (») cua Chung VI
Ky hieu
Bacillus subtilis
PO.03.B
LSD.01=1.49
CV (anh bdO-2,93%
LSD.01.2.51
CV (N) - 4,37%
LSD.01.2.73
CV(Fe)-3,15%
knuani
Ngudn carbon
Tinh bpt (0,5%)
68.45%
Tmh bpt (1%)
72.26%
Tinh bot (1,5%)
6li,93%
LSD.Ol -1,49
Ngudn nitrogen LSD.01=2.51 Ure (0.025%)
67,43%
72.87 73.69 70.43 76.55 70.87 74,23
Urd (0,050%) 69.13%
72,37 51.13 67.73 (54.71 72.31 79,52 25.71 174.i)3 74.93 ,1 (iS,93 67.52 70.88
l i r e (0.1)7.1%)
70.98%
72.40 J 66,81
67.64 73,43 64,49 74,61 74,96 75.11 69.37
Hwang vd cp (FeCU W FeCI, (0,25%) FeCl, (0,50%) FeCl, (0,75%) FeClj (0,25%) FeClj (0,50%) FeCI, (0,75%) FeCl, (0,25%) FeCl, (0.50%) FeCl,, (0.7,5%) Thay U 1^ ket ty = Z; ham lupng tinh bdt = X
Ham lupng ure ^ V cd dinh FeCl, d 0,75% se cd dupc
phuong trinh:
Z 117.012 100,7541*0,75 41,4311*X
101„32*Y + 13,264*0,75*0,75 + 119,49*0,75*X
22,5.347*X*X + 137,374*0,75*Y + 138,394*X*Y
5.2(i(i92*Y*Y - 180,288*0,75*X*Y
Dua vao phirong trinh hdi quy cd dinh bien Ham
lu^mg FeCl,, mat dap ung la:
Tir dd th| mat dap ung va dd thi dudng muc d
hinh 2 va hinh 3 dupc ve tir phirong dinh hdi qui khi
thay the khpang vd cp 0,75%, hnh bdt X (0.5%
-1.5%) va urd - Y (0.025% - 0,075%), cho thay 2 nhan td
linh bpt - X 1,036% va ure - Y = 0,04953% se cho
s,nh khdi vi khuan tdng hpp chat ket tu sinh hpc cap
nhat la Z- 79,53244%
:i!,«l2-i00flllW-4l,<3irj.:3'3n'-132iiTO'0;S-119«W
117.012 100.75410.75 41.4311'X
-Hinh 2 Dd thi mat d i p ung cua ty 1^ kit ta theo
khoing vd co = 0,75%, tinh bdt - X (0,5% -1,556) v4 ur6
= Y (0,025%-0,075%)
101,32-V + 13.264-0.75-0.75 +
119.49-0.75-0.65
0.55
0.45
0.35
0.25
il L 1
;
•-}
111
1 !
' / /
\
\
\
'-_
Function
— - 66,0 67,0 68,0
— 69,0
~ 70,0 71,0
—- 73,0 74,0
Hinh 3 Dd thj duAng miirccua t^lf kftta theo khoing vd
Y (0,025%-0,075%)
68
CO Fee,, = 0.7,5% hnh bOt - X (0,5% -1,5%) , , „ ,
NONG NGHIEP VA PHAT TRIEN NONG THON
Trang 7Tir cic kdt qua trdn chp thay khi bd sung 1% Unh
bdt, 0.05% urd va 0.75% khpang vd cp FeCl, vap mdi
tnrdng nudi sinh khdi ehung vi khuan Bacillus
siibtilis?O.03.B se eho ty Id kdt tu sinh hoc cao nhat
va ty 1^ thanh phan mdi tnrdng nay dupc su dung cho
cac thi nghidm nghien cuu tdi iru hda kha nang tdng
hpp chat kdt tii sinh hpc cda chung vi khuan Bacillus
subtilis PO.03.B va thii nghidm hieu qua xu ly nude
thai ca sd san xuat bun
3.1.4 Ndng dd dich vi khuan bd' sung vao mdi
tiwng kaohn
Ty le kdt tu ciia chung vi khuan Bacillus subtihs
PO.03.B cap nhat d ndng dp vi khuan 0.05% la 51,94%,
ndng dp 0.01% la 46,83% va thSp nhdt p ndng dp vi
khuan 0,02% (20,09%) Ty le kdt tu tang din theo su
tang dan ndng dp vi khuan, va khi a ndng dp cao
nhat a 0,05% thi su khac biet cd y nghia thdng ke
(Hinh 4) Dudng bieu didn bieu thi nhu hinh vdng
cung vdi dinh la 0,05%
5184 a
16.83 b H 14 71c
4d
Hinh 4 Anh hudng cua ndng dp dich vi khuan bd
sung Ain kha nSng kdt tu sinh hpc cua chimg vi
khuan Bacillus subtilis PO.03.B
Ndng dp dich vi khuan bo sung la 0.05% cho ty le
ket tu 51.94% ddi vo, chung vi khuan Bacillus subtilis
PO.03.B da dat hieu qua ket tu sinh hpc cap hpn sp
vdi cac nghien ciiu khac nhu: chiing Bacillus coagulants Ps 101 cd hiu lupng 40 ml/Ut cho d le ket
tu la 90% (Salehizadeh etal 2000), chung Bacillus hchenitormis Ueu lupng 150 ml/L cho d le kdt tu 98,4% (Shih etal 2001) 6 ndng dp 0.01% va 0,1% •
ty le ket tu khac biet cd y nghia thdng ke nhung de dam bao hieu qua ket nr va dd dang hong tinh toan thi ndng do vi khuan 0,05% dupc chpn d i thuc hidn thi nghiem tiep theo
3.2 Hidu suJit xir ly nude thai co sd sdn xu4t biin Kdt qua xu ly nude thai cp sd sa,] xuat bun cua
chung vi khuin Bacillus subtihs PO.03.B the hidn
hong bang 6 cho thay ham lupng BODj da giam 17,76%, chi tieu nitp long giam 21,7% va ham lupng ampnium giam 36,61% Ba chi tidu nay so vdi quy dinh nude dial dau ra cua QCVN_ 40/2011/BTNMT thi chi ed chi tieu nay ham lupng ampni dat cdn lai 2 chi deu kia khdng dat vi tipng thanh phan nude thai tir CO sd san xuat bun ehua nhidu chat him cp Tuy nhien chi tieu TSS giam 11,26% dat tidu chuan A nhung chi deu photpho long giam 21,67% chua dat tieu chuan loai B, nen cdn tich hpp cac phuong phap khae hay chung vi khuan bo sung de chi tieu nay dat quy dinh cua QCVN_ 40/2011/BTNMT vdi quy dinh nude thai diu ra
Qua cac chi tieu phan tieh khi ung dung chung
vi khuan tdng hpp chat ket tu sinh hpc Bacillus subtilis PO.03-B eho thiy ngoai kha nang kdt tu lam
g,am chi sd BOD, va TSS cdn lam g,am eac chi sd khac nhu n,tp tdng, photpho tdng va ham lupng amonium
Bing 6 Hidu suit xu ty ntrdc thai ca sd san
Chi tidu
BOD, (mg 0,/l)
TSS (nig/1)
Nita tdng (mg/l)
Photpho tdng (mg/l)
Ham Itrong amonium (mg/l)
.pH
Trude xu ly D/C
1520 36.40 179.2 17.07 11,2 4,05
xuit bun cda chung vi SauxulyPO.03.B 1493' 32,3' 140,0 • 13,37 • 07,1-4,35
khuan Bacillus subtilis PO.03.B
Ty le giam
%
17,76 11,26 21,87 21,67 36,61 7,50
QCVN 40 2011/BTNMT
A
75
50
20
4
5
6
B
150
100
40
6
10
11
Ghi chu:' Ket quit tiling binh 3 lan lap lal, phan ti'ch tai Phdng Thi nghiem chuyen sau - Trudng Dai hoc
Cin Tho'
69
Trang 8Mun chung cic chi tidu trong nuoe thai ca sd
san bun deu giam nhung chi cd 2 chi tidu TSS va
ham luprng amonium giam dat yeu cau do cd chi sti
thip B0D5 cao do boat dong cua vi khuin lactic
manh lam pH g,am Didu lam han che hoat ddng ciia
2 loai vi khuin ket tu sinh hpc chudng u pH cap (pH
9) ham lupng peohin va phpsphp trong hat gao
khdng cao nhung do sd iuon.g ldn (de lira biin) nen
lim gia tang chi so TX vi TP khi cap Chi vi khuan
kdt lu sinh hpc khdng the lam giam 2 chi tieu ma can
nhidu loai vi khuan chuydn hda \ vi tich luy poly-P
hd trpr, ndn vi khuan ke't m sinh hpc chi hoat ddng
trong giai doan dau cua quy trinh xu ly nude thai va
chinh pH nude thai cho phu hpp vpi chung
4.KtTUMN
Chung vi khuin Bacillus subtilis PO.03.B tdng
hpp chat kdt tu suih hpc chp hieu qua cap 79,5% d pH
9; bd sung mudi kim loai CaCl,; ngudn dinh dudng
dimg dd nudi smh khdi la tinh bpt 1,(W, ure (0,05%)
va khoang vd cp 0,75% (FeClJ va ndng dd dich vi
khuan 0,05% Hidu suit xir ly nude thai cp sd san xuit
bun chiing vi khuan Bacillus subtilisPO.03.1i sau 60
phut de lang chp ket qui cac chi tidu phan tieh ddu
giam: ham lupng BOD, (17,76%): TSS (11,26%): nita
tdng (21,87%): photpho tdng (21,67%) va ham lupng
amonium (36,61%) Trong dd, chi tieu TSS dat yeu
ciu Ipai A va ham lupng amoni dat loai B, cdn lai cac
chi tieu khac chua dat yeu elu theo QCVN 40
2011/BTNMT Kien nghi nghien cuu va bd sung
thdm cac dieu ki?n ung dung de tang hieu qua xu ly
nude thai
T H UEU THAM KHAO
1 Deng S R Bai and X Hu, 2003
CharacterisUcs of a biofloceulant produced by
Bacillus nwcilagiiiosus and its use in starch
wastewater Ueatment Appl niicrobiol biotechnol
60-588-593
2 Deng S Yu C , Ting V P 2005
Production of a biofiocculant by -Ispergillus
parasiticus and „s appbcation m dye removal Colloid
.^urf B Blomlerfaccs 44: 179-186
3 OrtJn.i; \V X S G Wang X F Sun X \\
Liu 0 Y Yue and B Y C7ao 20118 B,ofloi.culam
producUon by culture of Si'rraba I'lauia d„d ii>
application in wastewater treamieiit Bioresvur
Technol 99: lli;fUli;74
4 Kurane R K Haumiochi, 1 Kakunp, M Kivphara K Kawaguchi, Y Mizuno M H«ano and
Y Tamguchi 1994 Punfication and i,.,i,Kic,i/auon
of liqmd bioflpcculant prpduced by • ' • " " ' ' " " ' _ ' ' " '
erytiiropolis Biosci Biotechnol Biochem «
1977-19,12
5 Loan L T T V Phuong and C N Diep,
2018 Isolation and identification of bioflpcculant-producing bacteria, heterptrophic nittugen removal bacteria and pply-phosphate bacteria in wastewater irpm "My Tlio rice noodle" factories, Tien Giang province, Viemam International Journal of Innovations in Engmeering and Teclmolpgy (UIEniO (2): 185-198
6 U, W W., S Zhong H Y Lei, R W Chen, Q
Yu and H L Li 2009 Production of a novel
biofiocculant by Bacillus hcheniformis X14 and its
application to low temperatare drinking water
treatinent Bioresour Technol 100: 3650-3656
7 Salehizadeh, H., M Vossoughi and I Alemzadeh, 2000 Some mvestigatipns pn
bipflpcculant productipn bacteria Biochem Eng J, 5:
39-44
8 Salehizadeh, H and S A Shojapsadati 2001 Exti-acellular bippolymeric flocculants: recent trends
and biotechnologieal importance BiotechnolAdv, 19
(5): 371-385
9 Sathiyanarayanan, G., G S Kiran and J Selvin, 2013 Synthesis of silver nanoparticles by polysaccharide biofiocculant produced from marine
Bacillus subtilis MSBN17 Colloid Surf B Biolnterfaces 102:13-20
10 Sheng Y., Q Zang and H Wang, 2006 Sreening and flocculating of biofloeculant-produecing
microorganisms / Univ Sci Technol Beijing, 13:
289-292
11 Shih, I L, Y T Van, L C Yeh, H G Lin and
Y N Chang, 2001 Production pf a biopolymer
flocculant fi-om Bacillus hcheniformis and its flocculation properties Bioresour Technol, 78 (3):
267-272
12 Xia, S Q Z Q Zhang, X J Wang, A M
Yang, L Chen .! F Zliao D Leonard, N Jaffi-ezic-RenauU 2ll(l» l',-„duct,on and characc.,2ation of a
bi noctulani by Proleus mirabilis IJ-i Bioresour
Tedlnul Vi'y C-,T,2(yi',r>'Sl
'•' ''•™'-' " • • ' ' J - K i m S H Ahn, H R Lee -'"".• Chiuactenzation of a novel •.,-,ll„.vulant, p^
NONG NGHIEP VA PHAT TRIEN NONG THON - KY 1 - THANG
Trang 9KG03, from a marine dinoflagellate, Gyrodmium novel bipflocculant by multiple- imeroorgantsra
impudicumKCfB.BioresourTechnol,9S:Xl-'ieil consortia using brewery wastewater as carbpn
14 Ypkpi H., T Arima, S Hayashi, Y Takasaki, source JEnvnon Sci 19: 667-673
1996 Flocculation properties of poly(gamma- ,7 7,,.„„ ,- , , v v • r- „ „ ,
,, J J u o IT I , '-'• 'Jisng, >- Z L Ye, X L Fang Y H U and gutamic acid) produced by Bacillus subtihs J « , , , ^ , ,„„„ j , „ j u
' , „ 00 o., 07 W M Cai, 2008 Production and chai-aeter,st,cs of a
Femi^tBioeng,W.U.-%l biofiocculant produced by Bacillus sp F19
15 Zang J., Z Liu^^ S^ Wang and P Jiang 2002 5 , , , , , , ^ ^ y,^^^^,_ ^^^^^ g^^gg^
Characterization of a biofiocculant produced by the 18 Zouboulis A I Xiao-Li C, I A Katsoyiannis
manne myxobacterium Nannocysti's s^ NU-2 Appl 2004 The application of biofloceulant for the removal Microbiol Biotechnol, 59; 517-522 of humic acids from stabOized landfill leachates / 16.Zhang, Z Q., L Bo, X S Qing W X Jiang EnvmonManage.!^ {\):Z^,-^l
and Y A Ming, 2007 Production and application of a
OPTIMIZATION OF BIOFLOCCUIANT PRODUCED m BACILLUSSUBTJUSVO.QZ.BmD
ITS APPLICATION IN WASTEWATERTREATMENT FROM VERMICELU-NOODLE
FACTORIES
Le Tlii Loan', Cao Ngoc Diep^
' Microbiology Ph.D student
- Biotechnology R&D Institute, Can Tho University
Summary Wastewater from verraicelli-noodle factones contain a large number of organic especially residual starch and it must be treated before discharging into the environment BioflocculaUon is an extracellular polymer, which is produced by microorganisms It is safety, strong effect, biodegradable and hamilessness to humans and the environment in comparison to conventional synthesis flocculants, therefore they may be
researched and applied in wastewater vermicelli-noodle production sj'stem treatment Bacillus subulis
strain P0.03.B was isolated from wastewater of My Tho nce-noodle factories in Tien Giang provmce
Vietnam The optimal medium for Bacillus subtilis sb-ain 03.B consisted of starch (1.0%) urea (0.05%), and
FeClj (0.75%) at pH 9 with kaolin soluUon after 5 mmules together with CaCl solution and 0.05% inoculant (bactenal liquid) increased the flocculating activity up to 79.57% Applying this sU^ain for wastewater freatment in vermicelli-noodle factories, the results showed thai they reduced Biochemical Oxygen Demand (BODr), total solid suspension (TSS), total mtrogen, total phosphorus, and ammonium concenUBbons respectively 17.76% 11.26% 21.87%, 21.67% and 36.61% m companson to initial wastewater, respectively Especially, total sohd suspension and ammonium concentration parameters reached to A standard of QCVN_ 40/2011/BTNMT
Keywords: Bacillus subtilis, biofiocculant carbon source, nitrogen source, pH, wastewater
Ngireri phan bi^n: PGS.TS Le Diic
N g ^ nhin bai: 5/8/2019
Ngdy th6ng qua phan bien: 6/9/2019
Ngdy duyet dang: 13/9/2019