Báo cáo " Phân lập và khảo sát khả năng loại Hydrogen Sulfite (H2S) của vi khuẩn oxy hóa hợp chất Sulfur (SOB) trong nước kinh Nhiêu Lộc - Thị Nghè" docx

TAP
CHl KHOA HOC
VACONG
NGHE Tap
47,
s6
5,2009
Tr. 83-90
PHAN LAP VA KHAO SAT KHA
NANG
LOAI HYDROGEN
SULFITE
(H2S)
CUA VI KHUAN OXY HOA HOP CHAT SULFUR
(SOB) TRONG
Nu'ac
KINH NHIEU LOC - THj NGHE
NGUYEN
KHANH HOANG,
NGUYEN
HOAI
PHONG,
Hl/A
THACH
SON,
NGUYEN
VAN
CLFONG
I.MODAU
Nuac la mpt thanh phan rat quan trpng doi vai su song tren trai dat, day la dieu tat yeu ma
mpi nguai deu nhan thu'c dugc. Theo tac gia Timi Ecimovic (2005) be mat trai dat co dien tich

510,072
trieu km' trong do 70,7% dugc bao
phii
bai nuoc. Trong tong lugng nuac
ctia
qua dat
chi
CO
2,5% la nuoc nggt nhung phan lan (68,7%) nuoc nggt lai co mat trong cac
niii
bang. Vi
the,
lugng nuac ngpt con nguai co the de dang
sii'
dung dap trng nhu cau con lai rat it (0,4%)
trong tong so nuoc
ctia
trai dat.
Chat lugng
ciia
nuac co the str dung rat dang dugc mpi nguoi quan tam vi chting phai dap
ung dugc nhu cau
an,uong,
tam, giat. Tuy nhien, cac boat dpng
ctia
con nguai va thien nhien
hang ngay da thai vao nguon nuac rat nhieu hgp chat co the lam cho nguon nuoc tra thanh
khong an toan cho nhu cau sir dung
ciia
con nguoi va he sinh thai trong moi

trirong
nuoc. Mpt
trong nhung hgp chat thai ra do qua trinh boat dpng
ctia
con nguoi va thien nhien co anh huang
rat
Ion
den chi tieu mau sac va
miii ciia
nuac chinh la Hydrogen sulfite
(H^S).
Trong moi truang nuac cac hgp chat sulfur ton tai duoi 3 dang
tiiy
thuoc vao pH
ctia
moi
truang, khi moi truong co pH 6 thi 90% ton tai dang
H2S
va
10%
a
trang thai HS", nhung khi pH
moi truong dat gia trj
10
thi sulfur ton tai
100%
a trang thai
S"".
Hydrogen sulfite hoa tan trong
nuo'c voi nong dp toi da 0,4 gam/ 100 ml

a
dieu kien nhiet dp 20°C, khi c6 mat trong nuac
chting se gay
miii
hoi nhu
miii
trung thoi lam anh huong den gia trj cam quan.
Thanh pho Ho Chi Minh vai mang luoi kinh rach va song co tong dien tich be mat bao
phii
la 141 km" gom 5 he thong kinh rach:
Kinh Nhieu Lpc - Thj Nghe (NL-TN) bao
phii
33
km";
Kinh Tau
Hti
- Doi - Te bao
phii
37 km";
Kinh Tau Hu - Ben Nghe bao
phii 12
km";
Kinh Tan Hoa - Ong Buong- Lo Gom bao phu
16
km";
Kinh Tham Luong - Ben Cat - Vam Thuat bao phu 44 km .
He thong kinh rach
ctia
thanh pho Ho Chi Minh hien nay khong con kha nang tu lam sach
khi phai tiep nhan gan nhu toan bp lugng nuoc thai sinh boat va nuac mua

ctia
thanh pho dong
thai voi su lan chiem lam thu hep dien tich bao
phii ctia
he thong kinh rach dan den tinh trang 6
nhiem ngay cang tram trgng. Phan Ion he thong kinh rach trong pham vi npi thanh va khu vuc
dau nguon deu bi 6 nhiem nang va dugc
gpi
la "Kinh den" hoac "Kinh
thtii"
bai qua trinh
ehnyen
hoa cac hgp chat sulfate do nhom vi khuan khir sulfate (SRB) tao thanh hydrogen sulfite
igay
mill thoi va khi ket hgp voi sat tao thanh FeS gay mau den. Vi the, vao dau nam 2001 voi
sir
83
tai trg
ctia
ngan hang the gioi thanh pho da khai dgng du an ve sinh moi truang vai
mue
dich cai
tao kinh NL-TN va cai tai he thong thoat nuoc
ctia
thanh pho. Tuy nhien, hien nay du an van con
tiln hanh mac
dii
da dugc ngan hang the giai gia ban den quy I nam 2009.
Voi mong muon gop phan vao viec cai thien moi truong thanh pho
H6

Chi Minh cu
thi
gop
ph§n
cai thien chat lugng nuoc trong
h^
thong kinh rach, nhom nghien cuu da tiln hanh phan lap
vi
khuSn
co kha nang oxy boa hgp chat sulfur trong nuac kinh NL-TN sau khi da khao sat kha
nang loai hydrogen sulfite co trong nuac voi quy mo phong
thi
nghiem se tien hanh nghien ciru
CO
djnh nham tao ra che pham sinh hgc xtr li
H2S
trong co trong nuac thai sinh boat.
2.
NGUYEN LIEU VA
PHlTOTNIG
PHAP
2.1.
Nguyen lieu va hoa chat
- Mau nuoc thu nhan tir kinh Nhieu Lgc, Thj Nghe.
- Moi truong: PCA, PGA, Peptone, Colistine; Thuoc nhupm Gram (Difco); Bp test djnh
danh vi sinh vat: ID32E (bioMerieux).
- Til cay (Juoan, France).
- Noi hap (Hitachi, Japan).
- May do quang UV-VIS (Thermo Electron Cooperation, USA).
- Kinh hien vi dien ttr.

- Mo hinh thu' nghiem kha nang phan giai sunfite
ciia
vi
khuSn.
2.2.
Phu-QTig
phap
2.2.1.
Phucmg phdp ldy mdu vd chudn bi mdu
Hmh
1.
Cau Cong
Li
(cau Nguyin Van Tr6i)- Nai liy
miu
{Source:
www,
wikimapia.
org)
Jl
mau dugc lay tai kenh NL-TN a cau Nguyin Van
TrSi
co vj tri
(10''47'29"N;
106 40'55"E), do sau 0,1-0,2 m. Nhiet do cua mau nuac a khoang
26°C.
Mau dugc
\ky a
hai
miia

mua va
miia
kho, trong
mtia
mua (ttr thang 4 den thang
10)
va
miia
kho (ttr thang
11 din
thang 3) nam 2007
(bangl).
Mau nuac dugc dting dl phan tich ham lugng sunfite va phan lap vi
84
sinh vat. Nuoc dugc lay tai kenh va cho vao
erien
da triet triing giii'
a
nhiet do moi truang mang
ve phong thi nghiem. Mau dugc kiem tra trong thai gian 24 gid.
Bdng I. Chi tiet va thong so
ctia
mau nuoc
So
thii'
tu
1
2
3
4

5
6
7
8
9
10
11
2 ?
^ Phican
Lo
Nl
N2
N3
N4
N5
N6
N7
N8
N9
NIO
Nil
SPhdi
Thoi gian lay mau
Thang
11 -thang
3
Thang
11 -thang
3
Thang

11 -thang
3
Thang
11 -thang
3
Thang
11 -thang
3
Thang
11 -thang
3
Thang 4 - thang
10
Thang 4
-thang 10
Thang 4
-thang 10
Thang 4
-thang 10
Thang 4
-thang 10
;
phdn lap vi sinh
vgl.{V
Mua
Kho
Kho
Kho
Kho
Kho

Kho
Mua
Mua
Mua
Mua
Mua
Nhiet do
CO
26
25,5
26
26
26,5
26
24
24,5
25
25
25,5
larvey- Prescot, 2002):
pH
6.5
6,7
6,5
6,5
6,8
6,5
7,1
7,0
7,1

7,0
7,0
Thuy trieu
Len
Xuong
Xuong
Xuong
Len
Xuong
Len
Xuong
Len
Xuong
Xuong
Ba loai moi truong diing de phan lap vi khuan (moi truang Thiosulfate, pH = 8,5; moi
truang Beijerick, pH = 7,4; moi truang Thiobacillus thiooxidans, Ph = 4,4).
Sir dung pipette da triet triing
htit
5 ml mau vao ong nghiem chua san 20 ml dung djch moi
truong da triet trting. U
a 3TC
va theo doi trong thai gian
14
ngay. Hang ngay tien hanh cay khao
sat su phat trien
ctia
vi khuan
tii'
moi truong long sang moi truong ran. Moi truong ran dugc chuan
bj bang each them

1,5%
agar
(15
g /
1000
ml).
2.2.3.
Phu'ong phdp phdn lap vd ldm thudn de
chpn vl
khudn.
Str dung ki thuat
c4y
phan lap de tao khuan lac rieng le tren moi truang phan lap tuong
ling. Tang sinh vi khuan da phan lap tren moi truong tang sinh dang ran va dang long. Tien hanh
khao sat tinh
chSt
sinh hoa, hinh dang, kich thuoc, tinh chat bat mau bang
kT
thuat thir nghiem
sinh hoa, nhupm Gram.
2.2.4. Phicang phdp ldm tdng sinh vi khudn (Harvey- Prescot, 2002)
CSy
chuyin vi
khuSn
vao moi truang moi nham tang sinh khoi de phuc vu cong tac khao
sat kha nang xu li hydrogen sulfite. Cac moi truong tang sinh gom thach nghieng, dTa Petri va
moi truang long.
85
2.2.5.
Phuong phdp khdo sdt hdm

lucmg HjS
(APHA, AWWA and WPCF, TCVN- 4667- 88)
Nguyen tic: Phuong phap nay
thti'
nghiem kha nang lam giam
H2S
bai cac loai vi khuan
oxy hoa hgp chat sunfur
kit
hgp vai dung dich iode.
Tien hanh:
- Chuan bj dung djch iode: 20 g KI, 500 ml nuac cit, 5 g iode,
HCI
(1 : I); dung djch
NazSzOj
0,05N.
- Chuin dp miu blank: cho
10
ml dung dich iode va
5
ml muac cat va mpt vai gigt
HCI
vao
erlen
250 ml va
lie
diu.
dat cac
erlen
vao trong toi khoang

15
ml. sau do chuan do vai dung djch
Na2S203
0,1 N, sir dung chit chi thj la ho tinh bgt. The ticb
Na2S203
0,01 N chuan do dugc gpi
la
(Vo).
Lap lai thi nghiem nhu tren 3 lan.
- Chuin do vai mau thtr nghiem bao gom
H2S:
them 5 ml dung djch
H2S
vao erlen sau do
tiln hanh tuong tu nhu mau blank.
Ham lupng
H2S
dugc xac djnh theo cong
thii'c:
(F„-F,.).17.V,„^,^^^.1000
^H,S-
y
vai:
ning
do
Na2S203
la
A^^^
^^^
=

0,\N
; The tfch
H2S:
F^
^
= 5 ml; 17 la khoi lupng
(grams)
ciia
H2S;
1000
la the ticb pha loang;
Vx
la the ticb cua
Na2S203
0.05N dugc str dung de
chuan do 5 ml mau
H2S;
VQ
la the tich cua
Na2S203
0,05 N dugc str dung de chuan do 5 ml mau
blank.
2.3.
Khao sat kha nang
xir
li Hydrogen sulfite ciia vi khuan co trong moi
truong
- Mau nuac khao sat dugc lay tir kenh NLTN va tien hanh
xti'
li trong phong thi nghiem

duoi dang mo hinh be hieu khi. The tich mau nuac thtr nghiem la 50 lit vai so lugng vi khuan
khoang
lO^-lO"*
CFU/ml.Toc
do
khi
dugc
sue
vao khoang 10
lit
khi/giay nham
mue
dich co the
duy tri dugc mirc DO binh thuong vai dieu kien hieu khi tren 3
mg/l
(Frank A.Spellman.2003).
Thi nghiem thuc hien 3 lan
- Thu nhan mau nuac trong mo
hinh
sau moi gia va phan ticb bam lugng
H2S
bang phuong
phap
lot
dong thai xac djnh pH cua mau bang pH ke.
-Tien hanh thtr nghiem trong dieu kien khong bo sung vi khuan de doi chtrng.
3.
KET QUA VA THAO LUAN
3.1.
Phan lap vi khuan co trong mau nuo'c

Vi khuan oxi boa cac hgp chat sunfur dugc phan lap ttr mau nuac kenh NL-TN trong ba
loai moi truang gom
Mi
(pH 4,4),
M2
(pH 7,2) and
M3
(pH 8,5). Cac chting vi khuan nay co kha
nang str dung cac hgp chat sunfur (moc tren moi trucmg phan lap chuyen biet) va deu la
true
khuan gram am. Trong so cac vi khuin phan lap dugc chi 8 chung co kha nang oxi hoa cac hgp
chat sunfite (So lieu trinh bay trong bang 2). 8 chting vi khuin oxi boa bgp chat sulfite dugc
khao sat hinh dang bang kfnh biln vi dien ttr (hinh anh cua SOB
thi
hien trong hinh 2).
86
Hinh
2.
8 loai vi khuin
dugc
chup duai kinh hign vi - N3.1TT
(a);
N3.3TT (b); N3.4TT (c); N3.6TT (d);
N6.1TT(e);N6.2TT(f);N10,l K(g);Nll,l
K (h)
(Giai thich ki hieu N3.6TT: La vi khuan co so thu ty 6 dugc phan lap
Ur miu
nuac s6 3 tren moi truong
trung
tilth;

K
moi truong kiim)
Bdng 2. Tinh chat va so lugng vi khuan phan lap dugc tir mau nuac
STT
1
2
3
4
5
6
7
8
9
10
11
Lo
Nl
N2
N3
N4
N5
N6
N7
N8
N9
NIO
Nil
Ml
0
0

0
o
0
o
0
0
0
0
0
M2
o
0
X
0
0
X
0
0
0
0
0
M3
0
0
0
0
0
0
0
0

0
X
X
Miia
Kho
Kho
Kho
Kho
Kho
Kho
Mua
Mua
Mua
Mua
Mua
So vi khuan
phan lap
0
0
4
0
0
2
0
0
0
I
1
pH
6,5

6,7
6,5
6,5
6,8
6,5
7,1
7,0
7,1
7,0
7,0
Thuy trieu
Xuong
Len
Xuong
Xuong
Len
Xuong
Len
Xuong
Len
Xuong
Xuong
X. mau
CO
vi khuin
dugc
chon; O. mau khong co vi khuan
dugc
chon.
87

Bang 2 chi ra rang co 4 mau nuoc phan lap dugc vi khuin (2 miu
ctia miia
mua va 2 mau
ctia miia
kho). Co 2 mau nuac phan lap dugc vi khuan trong moi
trirang
trung tinh (M2), trong
khi do so con lai phat trien trong moi truong kiem (M3). Qua trinh phan lap vi khuin da chgn
dugc 6 chiing vi khuan ttr mau nuac
miia
kho va 2 chting vi khuin
tii'
mau nuoc
mtia
mua.
3.2. Kha nang khir cac hgp chat sunfur cua vi khuan
Trong nghien ciru khao sat kha nang xtr li hydrogen sulfite, nong do
H2S
c6 chi so trung
binh khoang 0,59 ppm va gia trj pH ctia mau niroc
a
dieu kien trung tinh (so lieu phan tich
khong dugc the hien trong bai bao cao). So lieu khao sat dugc trinh bay
a
binh 3 cho thay kha
nang oxi boa
H2S
cua vi khuan phan lap dugc tir mau. Sau 4 gia xu' If chting N3.4 dugc khao sat
cho ket qua xu If cao nhat ham lugng
H2S

giam 99,29%), trong khi do chting N3.3 dugc xu If cho
kit
qua thap nhat trong 8 chung dugc khao sat chi giam dugc 25%).
(%)
99.29
100.00
90.00
80.00
70.00
60.00
50.00
40.00
30.00
20.00
10.00
0.00
Removal
H2S
after 4 hours treatment period
(%)
• N3.4
83.OS
81.25 80.00
ONILI
73.44
• N10.1
• N6.1
es.is
54.84
N11.1 N10.1 N3.6 N3.1

• N3.6
25.00
N3.3
0
N3.1
• N6.2
1
• N3.3
Hinh
3.
Kha nang
xii'
li
H2S ciia
cac chiing vi khuan trong dieu kien mo hinh
Trong cac nghien cu'u
ctia
chting toi cac chting N3.4; Nl I.I; NlO.l; N6.I co kha nang phan
giai
HTS
tren
80%i
day la con so rat co y nghTa trong qua trinh lua chgn tmg dung vi sinh vat
trong sil' li moi truong vi hieu qua kha cao, trong khi do cac chimg khac kha nang xtr
If
duoi 80%
nhuN3.6;N3.1;N6.2,N3.3.
80
pH
7.8

7.6
7.4.
7.0
J
6.6
-I
6.4
-i-
pH Changing
Oh
1 h
2h 3h
pH
Times(h)
4
h
Hinh
4:
Gia tri pH sau 4 gia khao sat va xii li vai cac chung vi sinh vat phan lap dugc
88
Kha nang phan giai
H2S ciia
nhu'ng vi khuan phan lap dugc tir nuoc kinh NL-TN sau 4 gia
cho thay rang nhung san pham tao thanh khong lam giam pH moi truang. So lieu thi nghiem trong
hinh 5 cho thay gia trj pH trung binh trong qua trinh
xii'
If voi cac chiing vi sinh vat deu tang (so
lieu
ciia
tirng thf nghiem khong the hien trong bao cao).

4.
KET LUAN
Trong ket qua nghien ciru
ciia
chiing toi da phan lap dugc 8 chiing vi khuan co kha nang
khir
H2S
til' kenh NLTN.
Tat ca chimg deu la vi khuan hieu khi va la
true
khuan Gram am.
Trong 8 chimg nay co 4 chiing N3.4; Nl 1.1; NlO.l; N6.1 co kha nang loai
H2S
tren 80%
trong do
chimg N3.4
co kha nang loai
H2S
cao nhat la 99,29%.
San pham tao thanh
tii'
qua trinh oxi hoa hydrogen sulfite khong lam giam gia trj pH
ciia
moi truang (khong tao thanh axit)
Til'
ket qua nghien
cti'u
co the tiep tuc irng dung nhung chimg vi khuan co kha nang xir
li
H2S

cao bang kT thuat co djnh vi sinh vat nham tao ra sinh pham
xti'
If moi truong. Tuy nhien,
can phai tien hanh khao sat them kha nang xir If va cac
dae
tfnh sinh hpc
ciia
cac chimg vi khuan
nay de co the thuo'ng mai hoa san pham.
TAI LIEU THAM KHAO
1.
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wastewater and waste gases by biological conversion to elemental sulphur Colloidal and
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Physicochemical and
Engineering
Aspects 151 (1999)389-397.
2.
Alex T. Nielsen, Wen - Tso Liu, Carlos Filipe,
Leslie
Grady Jr., Soren
Molin,
and David
A.
Stahl
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Biological Phosphorus Removal Reactor, Applied and Environmental Microbiology 65 (3)
(1999)1251-1258.
3.
Altschul S. F., Thomas L. M., Alejandro A. S., Jinghui Z., Zheng Z., Webb M., and

Lipman D. J. - Gapped BLAST and
PSI
- BLAST: a new generation of protein databases
search
program.
Nucleic Acids Research 25
(17)
(1997) 3389-3402.
4.
Andrew S. Ball, David B. Nedwell and Rupert G. Perkins - Oxidation of hydrogen
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limicola.
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(2007)702-705.
5.
APHA AWWA and WPCF - Standard Method for Examination of Water and Wastewater
20th ed., Washington, D.C. American Public Health Association, 1998, p. 1325.
6. Aragno M. - Aerobic chemolithoautotrophic
bacteria.
In J. K. Kristjansson
(ed.).
Thermophilic bacteria, Boca Raton: CRC Press,
Fla,
1991, pp. 7-103.
7.
Bitton Gabriel - Wastewater Microbiology, Chichester: John Wiley & Sons, 3rd ed. 2005,
p.
768.
8. Brock T. D., and H. G. Schlegel - Autotrophic
bacteria.

Wis: Springer - Verlag, Madison,
1989,
pp.
1
-
15.
89
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(1989)189-221.
10.
Brune D. C. - Sulfur compounds as
photos\nthetic
electron donors. In R. E. Blankenship,
M. T. Madigan, and C. E. Bauer, Ano.xygenic photosynthetic bacteria, The Netherlands:
Kluwer, Dordrecht, 1995, pp. 847-870.
11.
Caroline M. O'Hara - Manual and Automated Instrumentation for Identification of
Enterobacteriaceae and Other Aerobic Gram - Negative Bacilli., Clinical Microbiology
Reviews 18(1) (2005) 147-162.
72.
Chirajyoti Deb, Ranadhir
Chakrabort}',
Amar N. Ghosh, Nitai C. Mandal, Tusharmouli
Mukherjee,
Pradesh
Roy - A generalized transducing thiophage (TPC - 1) of a facultati\e
sulfur chemolithotrophic bacterium, Bosea thiooxidans CT5, of a - Proleobacleria,
isolated from Indian soil, FEMS Microbiology Letters 227 (1) (2003) 87-92.
13.
Chris Easter, Chris Quigley, Peter Burrowes, Jay Witherspoon and Dirk Apgar - Odor and

air emissions control using biotechnology for both collection and wastewater treatment
systems.
Chemical Engineering Journal 113 (2005)93-104.
14.
Christoph Griesbeck et al - Mechanism of Sulfide - Quinone Reductase Investigated Using
Site - Directed Mutagenesis and Sulfur Analysis, Biochemistry 41(39) (2002)
11552-
11565.
15.
Dam Sao Mai, Nguyen Khanh
Hoang
Vu Dinh Huy, Abdukarimov
Abdusattor,
Abdurakhmonov Ibrokhim - The metal corrosion ability of sulfate reducing bacteria
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Dam Sao Mai, Nguyen Khanh
Hoang
Vu Dinh Huy, Abdukarimov Abdusattor,
Abdurakhmonov Ibrokhim - The biocide effect to sulfate reducing bacteria isolated from
the mine White Tiger, Petrovietnam journal Vietnam (5) (2007) 33-37.
17.
Daniel C. Brune - Sulfur oxidation by phototrophic bacteria, Biochimica et Biophysica
Acta
(BBA)
- Bioenergetics 97 (5)
(1989)
189-221.
SUMMARY
ISOLA FED

AND SURVEYED REDUCE HYDROGEN SULFITE
(H2S)
ABILLITY OF
SULFUR-OXIDIZFNG
BACTERIA (SOB) IN THE WATER
OF NHIEU LOC - THI NGHE CANAL
8 strains of Sulfur oxidizing bacteria (SOB) were isolated and tested in order to remove
hydrogen sulfide
(H2S)
from Nhieu Loc - Thi Nghe canal in the aerotank pilot scale. The
desulphurization ability of the strains
N3.4;
Nll.l;
NlO.l; N6.1;N3.6; N3.1; N6.2; N3.3 were
decreased (99.29%; 83.05%; 81.25%; 80.00%; 73.52%; 68.18%;
41.91%;
25.00%) respectively
Dia chi: Nhdn bdi ngdy 22 thdng 3 ndm 2009
Nguyen Khanh Hoang, Nguyin Hoai Phong, Htra Thach Son,
Truong Dai hpc Cong nghiep Tp.
H6
Chi Minh.
Nguyen Van Cuang,
Khoa
KT
thuat Y sinh, Truang dai hpc Chung Yuan, Dai Loan.
90