BO KHOA
HOC
VA CONG NGHE BO NONG NGHIEP VA PTNT
CHirONG TRINH
TRONG DIEM PHAT
TRIEN
VA
ITNG
DUNG CONG NGHE
SINH
HOC TRONG
LINH VlTC
NONG NGHIEP VA PHAT TRIEN NONG THON DEN NAM 2020
BAO CAO
CHUYEN
DE
KET QUA NGHIEN
ClTU
KHOA HOC CONG NGHE DE
TAI
Ten
chuyen de
PHAN LAP,
TUY6N
CHON,
DINH
DANH VA
DANH
GIA
CAC

KET QUA NGHIEN
ClTU
KHOA HOC CONG NGHE DE TAI
Ten chuyen de
PHAN LAP, TUYEN CHON, DINH DANH VA
DANH GIA CAC DAC TINH PROBIOTIC CUA CAC
•
CHUNG VI SINH VAT
H&U
ICH
Chu nhiem de tai Xac nhan nghiem thu cua
co*
quan chu
tri
TS.
Tran Quoc Viet
Ha Noi - 2010
Phanl
DAT VAN DE
Tu
thai
CO
dai,
con
nguai
da
biet Igi
dung nhung vi sinh vat c6
ich
>

thg
a con nguai va cai thien nang suat a vat nuoi mai chi
dugc
biet den trong mot vai thap ky tra
lai
day. Cac vi sinh vat
htJu
ich
dugc sii
dimg
cho muc dich nay dugc ggi la cac vi sinh vat probiotic va cac che pham
gom cac vi sinh vat huu ich dugc su dung nhu
nhiJng
chat bo sung trong thuc
an hoac nuac uong dung cho con nguai va vat nuoi dugc ggi la cac che pham
probiotic.
De san xuat dugc cac san pham probiotic, nha san xuat can c6 cac
vi
sinh vat huu ich, c6 day du cac dac tinh probiotic nhu c6 kha nang ton tai va
phat
triln
trong duang tieu hoa cua vat nuoi va con nguai, kim ham su phat
t
trien cua cac vi sinh vat c6 hai trong duang tieu hoa thong qua viec san
sinh
cac bacteriocin, axit huu ca, canh tranh duang chat va vi tri bam dinh tren
niem mac rugt vv. De c6 dugc cac vi sinh vat probiotic, can phai phan lap
chung tu cac
ngu6n
khac nhau trong tu nhien, sau do danh gia cac dac tinh

vuc nong
nghiep va
phat
trien
nong
thon "
giai doan 2006-
2009
Phan II
TONG QUAN TAI LIEU
2.1.
Khai niem probiotic
r r
Theo ngon ngu Hi Lap, probiotic c6
nghia
la "vi su song" Trong tieng
Anh,
CO
mot thuat ngu kha quen thugc, ngugc
nghia
vai tu probiotic do la tu
f
r
"antibiotic", tu nay c6 nghia la chong lai su song hay thuang ggi la khang
sinh.
Thuat ngu probiotic dugc Parker de nghi su dung lan dau tien vao nam
1974 de chi
"nhijng
vi sinh vat va nhung chat lam can bang he vi sinh vat
rugt" (Fuller, 1989).

sue
khoe
t6t
cho vat chu"
2.2.
He vi sinh vat ruot va
siic
khoe ciia he thong tieu hoa
6'
vat nuoi.
Hien nay, trong dinh duang gia
sue
hien dai,
sue
khoe cua he tieu hoa
dang la van de dugc ca the giai quan tam. Mot khau phan an
ly
tuang se
khong
CO
nhieu y nghia doi vai mot he tieu hoa khong
lanh
manh.
Sue
khoe
cua he tieu hoa a vat nuoi phu thugc vao rat nhieu yeu to, trong do su can
bang cua he VSV cong sinh c6 y nghia quyet dinh.
Khi con a trong bao thai, duang tieu hoa
ciia
vat nuoi a trang thai v6

VSV
Cac yeu to nay
chiu tac dgng cua moi truang, cua cac stress va tac dgng qua lai
lln
nhau.
Trong s6 cac nhan t6 tren, he VSV duang tieu hoa dong vai tro trung tam, chi
mot bien dgng bat
Igi
cua mot trong hai yeu to con lai cung anh huang xau tai
he VSV (Conway, 1994). Su cong sinh cua cac loai VSV trong duang tieu
hoa cua vat nuoi (chu yeu la trong rugt) tao nen mot he sinh thai ma va moi
can bang cua quan the VSV dugc xac lap chi mot
thofi
gian rat ngan sau khi
sinh (Jans, 2005).
Co nhieu quan diem khac nhau ve moi tuang quan can bang cua he
VSV rugt. Theo Jans (2005), de danh gia trang thai can bang, cac VSV rugt
dugc chia thanh 3 nhom (1) nhom chu yeu (main flora) gom cac loai vi khuan
ki khi
{Clostridia;
Lactobacilli;
Bifidobacteria;
Bacteroides,
Eubacteria);
(2)
nhom ve tinh (Satellite flora), gom chu yeu la Enterococci va
E.
Coli,
va (3)
nhom con lai (Residual flora) gom cac VSV c6 hai nhu Proteus,

sinh nhu: sinh ly vat chu, khau
phan thuc an va ca cau noi tai cua ban than he VSV. Thuc an la
n8n
dinh
7
\
7
X
duang ca ban cua
vi
sinh vat, bai vay su thay doi thanh phan khau phan, thuc
an khong dam bao ve sinh, phuang phap cho an khong hgp ly vv deu lam
ton hai den trang thai can bang he VSV rugt. Tuang tu nhu vay, cac chat bai
tiet cua he tieu hoa (dich mat, cac enzyme, chat dem va chat nhay vv)
cQng
nhu kieu va tan so nhu dgng rugt cung tac dgng
true
tiep den he vi sinh vat.
Kilu
va tin so nhu dgng rugt bi tac dgng rat lan bai cac stress (sinh de, cai
sfta,
don chuong, van
chuyen w).
Khi quan he can bang cua he VSV rugt bi
/
7
pha va se tao nen trang thai "dysbiosis" (trang thai "chung song c6 hai"). Bieu
7
hien cua trang thai "dysbiosis" a vat
chii

gia
sue,
gia cam non hoac trong dieu kien ve sinh kem va vat
nuoi chiu
nhieu
Stress. De vugt qua nhiing thach thuc do, da c6 rat nhieu nhung nghien cuu
nham tim ra
nhiJng
chat thay the khang sinh an toan vai vat nuoi. Mot trong
nhung chat do la probiotic.
2.3.
Cac vi sinh vat probiotic, vai tro va co che tac dong cua probiotic.
2.3.1.
Cac vi sinh vat probiotic.
Cac vi sinh vat probiotic thuang dugc su dung trong chan nuoi gom 3
nhom chinh: vi khuan (VK) lactic; VK Bacillus va nam men. Khi vao duang
tieu hoa cua vat nuoi thong qua thuc an hoac nuac uong, tuy theo loai ma ca
che tac dgng c6
Igi
cua chung
d8n
vat chu theo cac phuang thuc khac nhau.
Cac VK lactic (cac vi khuin chuyen hoa mot so loai duang nha qua trinh
f
r r
len men tao axit lactic) c6 kha nang san xuat cac chat (bacteriocin) uc che su
phat
triln
cua cac cac vi
khuin

Saccharomyces
boulardii) c6
kha nang
tmng
hoa doc to cua vi khuan (Castagluiolo va ctv,
1999),
gan ket
7
>
cac vi khuan c6 roi (phan lan la cac VK gay benh) nha su hien dien cua cac
1
t r
ca quan cam thu mannose va day chung ra khoi co the qua phan, nha do so
lugng cua cac VSV co ich tang len (Czerucka va Rampal, 2002).
Cac vi khuan va nam men thuang dugc su dung nhu
nhtjng
probiotic la
Bifidobacterium longum; B breve; B infantis; B
bifidum;
B adolescetis;
Lactococcus cremoris; L lactis; Streptococcus thermophilus; Enterococcus
feacium; Lactobacilus rhamnosus; L acidophilus; L casei; L bulgaricus; L
gasseri; Saccharomyces
boulardii;
S
cerevisiae.
2.3.2.
Vai
tro
cua probiotic

vi sinh vat duang rugt theo chieu huang co
Igi
cho vat chu; (2) tang cuang kha nang
miln
dich; (3) giam phan ung viem;
(4) ngan can su xam nhap va uc
ch8
su phat
triln
cua vi khuan gay benh; (5)
f
7
tang san xuat cac axit beo bay hai; (6) tang cuang qua trinh sinh tong hgp cac
vitamin nhom B; (7) tang hap thu chat khoang; (8) lam giam cholesterol huyet
thanh; (9) lam tang nang suat vat nuoi; (10) lam giam ham lugng amoniac va
r r
ure trong chat thai. Ngoai ra probiotic con rat an toan
v6i
dgng vat va than
thien vai moi
truofng.
Vi la chat bo sung VSV song
hOu
ich, viec
sii
dung
probiotic se khong tao ra cac chat ton du trong cac san pham chan nuoi co hai
cho sue khoe
ngucxi
tieu dung.

Salmonella w.
Mot so nam men probiotic (Saccharomyces
cereviese; S
Boulardii)
khong chi tranh vi tri bam dinh cua cac vi khuan khac
ma con gin
kSt
cac vi khuan co roi (phan lan la nhung vi khuan co hai) thong
qua cac ca quan thu cam mannose va day chung ra khoi vi tri bam dinh a
niem mac rugt (Czerucka va Rampal, 2002). Tuy nhien, canh tranh dinh
duang la phuang thuc canh tranh khoe
liet
nhat. Su sinh soi vai so lugng lan
r
cua mot loai vi sinh vat nao do la mot de dga nghiem trgng doi vai cac loai
khac
v6
nguon ca chit cho phat trien.
D6ng
thai vai canh tranh loai
trir,
cac VSV probiotic con san sinh cac
chit kim ham vi khuin nhu lactoferrin, lysozyme, hydrogen peroxide cung
nhu mot
s6
axit
hiJu
ca khac. Cac chit nay gay tac dgng bat
Igi
len vi khuin

probiotic doi vai viec nang cao
chuc nang miln dich van con chua dugc hieu biet day
dii.
2.4.
NhCrng
dac tinh probiotic cua cac chung vi sinh vat huu
ich.
7 7
> >
De
CO
the dugc su dung nhu nguon probiotic, cac VSV can hoi
dii
cac
dieu kien sau: (i) phai la cac vi sinh vat
hiJu
ich, v6 hai doi vai vat nuoi va
con nguai; (ii) song dugc trong duang tieu hoa
ciia
vat nuoi vai cac dieu kien
kha bat
Igi
(pH thay doi tu rat toan den kiem, muoi mat va cac dieu kien sinh
hoa trong lumen rugt w); (iii) co kha nang uc che cac VSV co hai bang
nhieu
each
khac nhau (canh tranh loai tru, san sinh ra cac chat uc che, tang
cuang dap ung mien dich
w);
(iv) c6 the sinh truang va phat trien trong cac

(ciing
giong,
cung loai nhung dac tinh probiotic khac nhau rat lan giua cac chiing).
Phuang phap xac dinh danh phap
ciia
cac chiing VSV probiotic phai dugc
chung thuc bing ten khoa hgc dugc cac
t6
chuc chuyen mon co uy tin chip
nhan va mang tinh
ph6
biln.
Mudn phat buy tac dung, cac VSV probiotic phai ton tai dugc trong
duang tieu hoa
ciia
vat nuoi,
kh^
nSng
nay khong
hi dl
dang
d6i v6i nhilu
loai VSV ma moi
tru5fng
cu tru thuang xuyen
ciia
chiing khong phai la trong
he tieu hoa
ciia
dgng vat cap cao. Bai vay, dl chgn

vat nuoi. Mat
khac quan he can bang
ciia
he vi sinh vat rugt a nguai va dgng vat la mot he
\ \
7
can bang dgng, nen viec danh gia kha nang ton tai va phat trien
ciia
cac chiing
dugc
lira
chgn trong duang tieu hoa
ciia
vat nuoi cang tra nen can thiet (Jan,
2005).
Kha nang ton tai
ciia
cac chiing VSV probiotic trong duang tieu hoa
ciia
vat nuoi dugc danh gia thong qua cac thi nghiem in-vitro nham khao sat
kha nang song trong moi truang co do pH khac nhau, kha nang bam dinh vao
niem mac rugt (Misha va ctv, 2005) va kha nang chiu muoi mat (Dora va ctv,
2003).
Cac
thii
nghiem in-vitro rat huu ich de thong qua do hilu dugc khong
chi nhung dac tinh sinh hgc ma con cac ca che tac dgng
ciia
cac chiing VSV
dugc

kl d6i
vai hoat tinh
ciia
chiing. Rit may min la chi c6 mot
s6
thanh phin co hoat tinh nhu mot s6
mu6i kim loai, cac chit axit hoa, mot
s6
chit
b6
trg khac la co tac dgng tieu
cue
den
sue s6ng
cua cac chiing VSV probiotic, con phin lan cac thanh phin
khac
ciia
khiu phin
diu
v6 hai
d6i
vai chiing.
Vl
nguyen tic, gia
trj
probiotic
ciia
mot chiing VSV probiotic ty le thuan vai tinh tuang thich cua chiing
d6i
vai cac chung khac vai cac thanh phin

TT
1
2
3
4
5
6
San pham khoa hoc cua
dS
tai
Ten san pham
Che pham
probiotic cho
Ign
va ga
Che pham da
enzyme tieu
hoa
Qui trinh san
F
Ft
xuat che pham
probiotic, che
t
pham da
enzyme
Mo hinh sur
dung probiotic
va che pham da
enzyme

ty le benh duang tieu hoa:
8-15%
+Hoat
tinh
-Amylaza: 1000
lU/g
Proteaza: 100
lU/g
-
p-Glucanaza
:
200
BGX/g
Xylanaza:
1000
lU/g
-Xellulaza: 1000
lU/g
+Tang sinh truang: 6-
10%
F
w
+Giam tieu ton TA: 5-
10%
Qui mo san xuat: 20-50 lit/me;
25-50 kg /me
- Dugc ap dung tai ca sa san xuat
-
1 -2
mo hinh chan nuoi

dugc hai san
ph§m
sinh hgc (probiotic va
enzyme tieu hoa) dung trong
chan
nuoi.
M6i chi phIm
co nhung dac
dilm
khac nhau
v6
thanh phin, hieu qua tac dgng doi vai vat nuoi, phuang
thiic
san
F
xuat va su dung. Bai vay, ngi dung va phuang phap nghien cuu
ciia
de tai
dugc chia thanh hai
ph^n
rieng biet. Chuyen de nay he
thdng
cac ket qua
nghien
ciiu
lien quan
d8n
viec phan lap, danh gia va tuyen chgn cac vi sinh
vat huu ich nhu ngudn probiotic phuc vu cho viec san xuk cac
chi phIm

4.3.
Vat lieu va
phu-ong
phap nghien
cihi
4.3.1.
Vat lieu nghien
cwu.
- Cac vat lieu phuc vu cho viec phan lap
tuyin
chgn cac vi sinh vat probiotic:
Phan, chat
chiia
ducmg tieu hoa (rugt non, manh trang)
ciia Ign
va gia cim
12
(ga, ngan, vit). Mot
s6
san phim probiotic thuang mai (Lacto-Sac
ciia
hang
Alltech-Canada; EM
ciia
Nhat; E-Lac
ciia
Han qu6c vv)
- Cac vi sinh vat
kilm
dinh {Salmonella enteritidis, E. coli va Shigella

a
n6ng
do tu
10'^-10"^
va ciy gat tren moi truang MRS
(d6i vai vi khuin Lactic) va YM
(d6i
vai nim men). Sau khi ciy cac
mlu
dugc giu trong
tii
im a nhiet do
37°C
cho vi khuin (VK) va 30
°C
cho nim
men trong 48 gia.
- Sau 48 gia tren moi truang MRS chgn cac khuin lac tao vong phan giai
CaCOs;
tren moi truang (Yeast Malt- YM) chgn cac khuin lac nim men tron
deu, tran nhan soi kinh co hinh trung va nay choi.
4.3.2.2. Danh gia dac diem hinh thai, phan loai vd dinh danh cac chung VSV
hitu
ich.
-
Hinh
thai t6' bao dugc quan sat sau khi nu6i
c^'y
tir
24 den 48

13
Viec phan loai cac chung vi khuin dugc thuc
hi6n
dua tren ca
sd
khoa phan
loai cua Bergeys (Holt va ctv, 2000) (theo cac
chi
tieu chii yeu nhu:
hinh
thai
te'
bao, khuin lac,
nhu6m
Gram, phan
liiig
catalaza va
len
men mot so loai
dudng)
va phan
tich,
giai trinh tu 16S rARN theo phuang phap dugc
gi6i
thieu
bc^i
Lane (1991).
- Xac dinh trinh tu rADN 16s
ciia
cac chiing vi khuin lactic theo phuang phap

m6i trucmg dich the trong dem acetat co pH khac nhau (2,2; 3,0; 4,0; 5,0; 6,0;
7,0), tren may
lie
6n nhiet (200 vong/ phut tai 37°C cho vi khuan Lactic va
30°C cho na'm men),
- Danh gia kha nang sinh truang trong moi trudng co nong do muoi mat khac
nhau: Cac chung nghien
ciiu
cGng dugc nuoi
c^y
tren m6i trucmg dich the nhu
tren
CO
nong
d6
muoi mat (0,2; 0,5;
1;1,5;
2; 3%). Nu6i cay
lie
(200
vong/phut tai 37°C doi
v6i
vi khuan va 30°C d6'i v6i
na'm
men).
De'm
mat do te' bao sau 48h nu6i ca'y gat tren
dia
Petri c6
chiia

Vi sinh vat (vi khuin va na'm men) dugc nu6i ca'y
thu^n
khiet trong m6i
trucmg thich hgp cho
tirng
chiing.
Chuin bi cac mau ruot ga tuai:
Rua cac doan
ru6t
non (tii ga broiler manh khoe) 3 lan vai dem PBS sao cho
ta't ca cac vi sinh vat
khdng
con tren hi mat niem mac
ru6t.
Rua 1 lan vai moi
trucmg (MRS cho vi khuin va YM cho na'm men).
Tien hanh thu: bam
dinh:
-
Phii
dich te bao da chuin bi a tren len tren be mat niem mac
ru6t.
- U 90 phut trong tii am
(37°C).
- Rua
mau ru6t
\in 1 bing
nudfc
muoi sinh
lyl%.

(MRS cho VK Lactic; thach
thudng
cho VK Bacillus va YM cho na'm men)
15
nhung
CO
b6 sung cac thanh
phSn
co hoat
tinh
thucmg co trong cac khiu
ph^n
an cho
Ign
va gia
cSm
gom
m6t
s6' loai khang sinh: Bacitracin Methylene
Disalicylate (BMD) (50 ppm); Saigon Nox (100 ppm), Colistine 98% (100
ppm),
chlotetracyclin (CTC) 15% (100 ppm);
m6t
s6' loai khoang CuS04 (250
ppm Cu); ZnS04 (100 ppm Zn) va h6n hgp axit
hiJu
ca (g6m axit lactic axit,
axit formic, axit citric.)
v6i li^u
200mg/lit

san
phim probiotic thuang mai), 64 chiing vi sinh vat da dugc phan lap, trong do
CO
27 chung vi khuan lactic (42,2%) va 39 chung na'm men (57,8%) (bang 1).
Bang
1:
Ke't
qua phan lap cac vi sinh vat tii cac nguon khac nhau
Ngu6n phan lap
Chat
chiia ru6t
non va
ru6t
gia cua
Ign.
Cha't
chijfa ru6t
non,
ru6t
gia cua ga
B6'n
(4)
mau
san phim probiotic thucmg
mai
T6ng
Vi sinh vat
VK Lactic
10
15

cho tha'y, ta't ca cac san phim nay diu la cac san phim probiotic da chung. De
lam ca
sd
cho viec tuyen chgn, cac chiing VSV tren tie'p tuc dugc
thu^n
khie't
tren
m6i
trucmg
dia
thach va chgn dugc 4 chiing (2 chung VK lactic va 2
chiing na'm men) de danh gia cac dac
tinh hinh
thai, sinh ly, sinh hoa quan
trgng lam ca
sd
cho viec phan loai. Cac ke't qua dugc trinh bay
d
cac bang 2 va
3.
Bang 2: Dac dilm
hinh
thai, sinh ly, sinh hoa ciia 2 chiing vi khuin lactic
TT
1
2
3
4
5
6

Riboza
Xyloza
Arabinoza
Rhamnoza
Trehaloza
Lactoza
Mannitol
Sucroza
Cellobioza
Raffinoza
Galactoza
Tinh bgt
Gluconat
Fructoza
Thanh
te'
bao co meso-DAP
+
-1-
+
+++
-I-+
+++
+++
++
+
-1-
++
+
-1-

ciiu trir
tinh
b6t,
trong
do chiing
NCI
co kha nang d6ng hoa tinh
b6t or miic
th^p,
nhung lai khong co
17
kha nang d6ng hoa riboza va xyloza. Ca 2 chiing deu co kha nang dong hoa
t6't 2 ngu6n carbohydrate ca ban la Sucroza va Cellobioza.
Hai chung na'm men
d^u
co
hinh
tiling,
elip,
nhung SC nay ch6i 1,2,3
phia,
trong khi do SB chi nay ch6i
1
phia.
Ca 2 chiing diu co
hinh
thai khuin
lac nhu nhau (tring siia, tron nhan bong va c6 vong d6ng tam) (bang 3).
Bang 3:
M6t

bao
Hinh thdi khudn
lac
Sorboza
Riboza
Xyloza
Arabinoza
Rhamnoza
Trehaloza
Lactoza
Sucroza
Cellobioza
Raffinoza
Galactoza
Tinh b6t
Melibioza
Glucoza
Sinh axit
Phan ling DBB
Phan ling thuy
phan ure
Chiing nam men
Saccharomyces
cerevisiae
(SO
Hinh
trumg,
eh'p, nay ch6i
1,2,3
phia

+
-
+
-
+
+
-
-
+
-
Saccharomyces boulardii
(SB)
Hinh
trutig,
eh'p, nay ch6i 1
phia
TrSng
sua tron nhan bong
loi
CO
vong d6ng tam
Khd nang
dong hod
nguon
cacbohydrat
-
-
-
-
+

ciia
Bergeys va cac du lieu
b
cac
bang 2 va 3
thi
chiing
NCI thu6c
nhom Lactobacillus fermentum, chiing NC2
thu6c
nhom Lactobacillus casei, chiing SC
thu6c
Saccharomyces cerevisiae
va chung SB
thuOc
nhom Saccharomyces nhung chua ro loai. Tuy nhien, de
ggi ten chinh
xdc,
cac chiing tren da dugc giai trinh tu ARN va ADN
16S.
Cac
ke't qua dinh danh dugc trinh bay nhu sau:
Trinh tu rADN
16S
cua chiing
NCI
>NC1
GCCAACGAGTGGCGGACGGGTGAGTAACACGTAGGTAACCTGCCCAGAAGCG
GGGGACAACATTTGGAAACAGATGCTAATACCGCATAACAGCGTTGTTCGCAT
GAACAACGCTTAAAAGATGGCTTCTCGCTATCACTTCTGGATGGACCTGCGGT

chiing NCI tuang d6ng vofi trinh
tu
rADN
16S
ciia
Lactobacillus fermentum
99,7 % (1446/1450).
Trinh
tu
rADN
16S
ciia chiing
NC2
>NC2
ACGCTGGCGGCGTGCCTAATACATGCAAGGCGAACGAGTTCTCGTTGATGATCGG
TGCTTGCACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTG
GGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCA
TAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTT
GGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGA
TGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGC
CCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCT
GATGGAGCAACGCCGCGTGAGTGAAGAAGGCTGTCGGGTCGTAAAACTCTGTTG
TTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCAACCA
GAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGC
GTTATCCGGATTTATrGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATG
TGAAAGCCCTCGGCTTAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTG
CAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGA
AGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGA
AAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGAT
GAATGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTAAG

SC
>SC-I1
TTAAAGAAATTTAATAATTTTGAAAATGGATITrTrTGTTrTGGCAAGAGCATGA
GAGCmTACTGGGCAAGAAGACAAGAGATGGAGAGTCCAGCCGGGCCTGCGCT
TAAGTGCGCGGTCTrGCTAGGCTTGTAAGnTCnTCrTGCTArrCCAAACGGTGA
GAGATrTCTGTGCTrTTGTTATAGGACAArrAAAACCGTTTCAATACAACACACT
GTGGAGTnTCATATCnTGCAACi-lTnClllGGGCATTCGAGCAATCGGGGCCC
20
AGAGGTAACAAACACAAACAATTTTATTTATTCATTAAATTnTGTCAAAAACAA
GAATTTTCGTAACTGGAAATTTTAAAATATTAAAAACnTCAACAACGGATCTCT
TGGTTCTCGCATCGATGAAGAACGCAGCGAAATGCGATACGTAATGTGAATTGC
AGAATTCCGTGAATCATCGAATCTTTGAACGCACATTGCGCCCCTTGGTATTCCA
GGGGGCATGCCTGTTTGAGCGTCATTTCCTTCTCAAACATTCTGTTTGGTAGTGAG
TGATACTCITTGGAGTTAACnTGAAATTGCTGGCCTTTTCATTGGATGTTTITnTC
CAAAGAGAGGTTTCTCTGCGTGCTTGAGGTATAATGCAAGTACGGTCGTTTTAGG
TTTTACCAACTGCGGCTAATCnTTTTATACTGAGCGTATTGGAACGTTATCGATA
AGAAGAGAGCGTCTAGGCGAACAATGTTCTTAAAGTTTGACCTCAAATCAGGTA
GGAGTACCCGCTGACTTAGCATATCAATAGCGAAGAAAGAAAGCATATCAATAA
GCGGAGGAAAAGAAACCAACCGGGATTGCCTTAGTAACGGCGAGTGAAGCGGC
AAAAGCTCAAATTTGAAATCTGGTACCTTCGGTGCCCGAGTTGTAATTTGGAGAG
GGCAACnTGGGGCCGTTCCTTGTCTATGTTCCTTGGAACAGGACGTCATAGAGG
GTGAGAATCCCGTGTGGCGAGGAGTGCGGTTCTTTGTAAAGTGCCTTCGAAGAGT
CGAGTTGTTTGGGAATGCAGCrCTAAGTGGGTGGTAAATTCCATCTAAAGCTAAA
TATTGGCG
AGAGACCGATAGCGAACAAGTACAGTGATGGAAAGATGAAAAGAACTTTGAAA
AGAGAGTGAAAAAGTACGTGAAATTGTTGAAAGGGAAGGGCATTTGATCAGAC
ATGGTGTnTGTGCCCTCTGCrCCTTGTGGGTAGGGGAATCTCGCATTTCACTGGG
CCAGCATCAGnTTGGTGGCAGGATAAATCCATAGGAATGTAGCTTGCCTCGGTA
AGTATTATAGCCTGTGGGAATACTGCCAGCTGGGACTGAGGACTGCGACGTAAG
TCAAGGATGCTGGCATAATGGTTATATGCCGCCGTCTTGAAACAGGACC

v6i Saccharomyces cerevisiae
Cac ke't qua
phan loai theo phuang phap
c6
diln
va dua vao
phan tich trinh
tu
ADN va ARN
dugc trinh
bay
b bang
4.
21
Bang 4:
Kit
qua phan loai va dinh danh cac chung dugc lua chgn
STT
1
2
3
4
Kihieu
NCI
NC2
SC
SB
Dua vao phan tich trinh tu
16S
rARN

va Lactobacillus
casei-NC2.
5.2.
Dac tinh probiotic ciia cac chiing VSV
hiifu
ich.
Cac chiing vi khuin Lactic
thu6c
nhom L. fermentum va L. casei cung
nhu cac chung na'm men
thu6c
cac nhom S. cerevisiae vd S. boulardii la
nhiing chiing VSV lanh tinh (khong phai la nhiing VSV gay benh), thucmg
dugc
sir
dung nhu nguon VSV Probiotic (Sanders va ctv, 2001). Tuy nhien,
m6i m6t
chung thugc cac nhom nay co cac dac
tinh
probiotic manh yeu khac
nhau.
Bcri
vay,
tru6c
khi dugc su dung, cac chung nay
cdn
dugc danh gia cac
dac tmh probiotic. Ke't qua danh gia dugc trinh bay
6
cac bang 5; 6; 7; 8 va 9.

tiT
2-4 x
10'
cfu/ml or
55°C).
Tuang ling
vdi
su giam sinh khd'i VSV, ham lugng axit lactic cung giam
manh. Dap
ling
vl sinh trucmg doi vdri nhiet
d6
nu6i ca'y cua 2 chung na'm men
(SC va SB) ciing co xu
hu^g
tuomg tu
dd'i
vdri nhom VK lactic, nhung 2 chiing
nam men to ra sinh trucmg tot han khi nu6i ca'y or nhiet
d6
30°C.
Ngoai
tr^
nhihig
VSV dugc phan lap tii m6i truomg ben ngoai, con hiu het cac VSV
22