Trng i hc Nha Trang Vin CNSH &MT

Viable counts, characteristic evaluation for commercial lactic acid bacteria products
GVHD: L Phng Chung Lp : 52CNSH Nhm : I

NI DUNG
Abstract
1. Introduction 2. Materials, methods and results 3. Discussion

TM TT (Abstract)

TM TT
Kim nghim 8 sn phm probiotic thng mi ca chng vi khun lactic acid( LAB) S lng t bo Chu acid, dch mt Kh nng bm dnh vo t bo biu m rut Kh nng khng cc nhn gy bnh

Kt qu:
Mt t bo trong cc sn phm cht lng cao hn cc sn phm rn. 2 sn phm LAB dng rn, s lng LAB sng st khng ng nh thng tin trn bao b Cc chng LAB t cc sn phm thng mi khc nhau c kh nng thch nghi vi acid v dch mt khc nhau Phn lp c 25 chng c kh nng khng cc tc nhn gy bnh ng rut Trong 25 chng th ch c 5 chng c kh nng bm vo biu m rut

Kt lun
Ch c vi sn phm LAB thng mi mi p ng yu cu c bn v chc nng ca probiotic.

GII THIU (Introduction)

Gii thiu
Ci thin sc khe H tr ngi b ri lon tiu ha v vim rut cp tnh Ci thin chc nng tiu ha

Sn phm LAB
Dng lng: sa chua

Gm 2 dng
Dng rn: Bt, ht nh, vin nn

Sn phm LAB
Hai hoc nhiu chng LAB c kt hp s dng trong mt sn phm Gm cc chng:
Lactobacillus thermophilus Streptococcus bulgaricus L. acidophilus L. casei Bifidobacterium longum

iu kin vi khun lactic c s dng lm ch phm sinh hc:

An ton Kh nng tn ti khi ch bin v bo qun Chu c acid v dch mt Khng tc nhn gy bnh Bm vo t bo biu m rut

VT LIU, PHNG PHP V KT QU (Materials, methods and results)

Sn phm thng mi LAB

Kho st s lng t bo sng

Kho st s lng t bo sng

Mi trng MRS: (De Man, RoGoSa, Sharpe ) Mi trng BCP: (Bromo Cresol purple)

Quy trnh nui cy

Nghin 10g dng vin nn hoc dng ht Ly 1g bt hoc 1ml sa chua

Dung dch: 9ml m phosphate PBS, pH =7.2

 Tin hnh pha long dung dch 10 ln lin tip vi PBS 1/10 1/10 1/10 1/10 1/10

37 0 C trong 48h( mt MRS) or 72h( BCP)

1 ml

KT QU

Kho st t l sng trong iu kin acid v dch mt

Kho st t l sng trong iu kin acid

1ml sa chua hoc 1g bt

9ml PBS pH=2

9ml PBS pH=2.5

9ml PBS pH=3.2

9ml PBS pH=7.2

37oC, 3h.

Kho st t l sng trong iu kin acid

Mi trng MRS, k kh 37oC, 48h

Kim tra nh hng ca dch mt n s pht trin ca t bo LAB

Ly tm 7000rpm, 5 pht Ra PBS + 10ml MRS

LAB pH=2

C hoc khng 0.3% mt b

37oC , 6h K kh

Kh nng chu mt b ca LAB c xc nh bng cch so snh vi cc t bo LAB sng trong mi trng thch MRS

KT QU

Chn lc v phn lp (Selection and identification)

Mi sn phm, chn ngu nhin 5 khun lc ri. C 25 chng c chn t cc sn phm LAB Test sinh ha
Catalase (-) Gram dng Khng di ng Hnh que( rod) hoc dng cu khun

Kho st hot tnh khng khun (Antibacterial activity assay)

Kho st hot tnh khng khun (Antibacterial activity assay)

S dng phng php khuch tn mi trng thch (Toure v cng s (2003) ,Garriga v cng s (1998) Cc chng ch th bao gm - Escherichia coli (ATCC 11.775) - E. coli sinh c t( enterotoxigenic) (ETEC) (ATCC 43.886) - Salmonella typhimurium (ATCC 14.028 v chng I50) - Staphylococcus aureus (ATCC 9144).

Chng ch th Pha long

100l

To 3 ging

Ly tm 7000rpm/5pht Nui cy LAB

100l

Kt qu
Vng c ch < 10 mm : khng c ch (-) > 10 mm : c ch yu (+) > 15 mm : c ch mnh m (+ +).

Mi kho st c thc hin trong ba ln.

S khng li vi khun gy bnh ca t bao LAB

T bao LAB c ngun gc t cc sn phm A, B v D thi c ch (inhibitory) tt c cc chng ch th. T bao LAB co ngun gc t cc sn phm t C v E khng c ch cc chng ch thi tr S. aureus (khng biu hin kt qu).

Nu cc t bo LAB c th nhanh chng i qua d dy v c tru trong rut, sn phm ca chng trong mi trng acid c th c ch s pht trin ca cc vi sinh vt c hi.

Kho st s bm gn ca LAB (Adherence assay of LAB)

Nui cy t bo biu m Int-407

Int-407( ATCC CCL 6) l t bo ngi c ngun gc t rut v hi trng ca phi ngi. Nui cy t bo n lp kho st

Nui cy t bo biu m Int-407

1

Int-407 nui cy trong mi trng BME, b sung 15%(v/v) FBS. 37oC, 5% CO2 Thu t bao sau 3 ngay. Ra t bao 2 ln vi PBS, tch t bo bng trypsin 0.05% sau o cy truyn sang a 24 ging c mi trng nui cy v trng cho n khi cc t bo pht trin n lp trong mi ging.

Kho nghim s bm gn ca t bo LAB

Chun b t bao LAB: ra 2 ln vi PBS v ly tm trong 5 pht vi 7000rpm, sau o c ha tan trong 1ml BME

Chuyn tb LAB (khong 108 tb/ml) vo a 24 ging cha t bo Int-407 n lp, 370C, 5% CO2, 2h.

. gn ca t bao LAB Kho nghim s bm

Ly cc t bo Int-407 trong 15 ging c chn ngu nhin ra 2 ln vi PBS, trn vi formallin 10% trong 30 pht, nhum vi thuc tm trong 5 pht, ra li 3 ln vi PBS, ri quan st di knh hin vi ( phng i gp 200X).

m t bo LAB trn khong 100 t bo Int-407 xc nh s lng t bo LAB b dnh cht vo t bo Int-407.

Cc t bao LAB bm dinh trn b mt t bao Int407

V sao phi lm test ny?

Cc chng probiotic bm dnh c vo thnh rut , khu tr tt trong ng tiu ho sinh si ny n.
c ch vi sinh vt gy bnh, bo v biu m, tng kh nng min dch ca vt ch.

Tng kh nng cnh tranh ca cc chng probiotic vi cc vi sinh vt bt li khc.

Xc nh, nh danh vi khun t sn phm B

5 chng LAB thu c t sn phm B cng mc bm dnh, khng khun 5 chng c s i khng (antagonistic) ging nhau vi vi khun ch th. 5 chng cng hnh que. nghi ng 5 chng thuc cng mt loi.

Phng php sinh hc phn t

S dng b kit API 50 CHL

Phn tch NST DNA bng phng php in di gii trnh t.

Kt lun cng mt loi l L.acidophilus

THO LUN

Tho lun
Sn phm probiotic LAB l mt trong nhng thc phm quan trng trn th gii iu kin chng LAB c s dng lm ch phm

Lactobacilus acidophilus

 Hu ht sn phm LAB thng mi c tuyn b c s lng LAB cao, hu ch cho chc nng probiotic

 Mt s sn phm probiotic c tuyn b cha chng LAB c trng, co th tn ti va th hin chc nng ca chng trong ng tiu hoa.
S dng t bao Int-407 nh gi t l sng st, kha nng bm dinh trn b mt t bao biu m rut va kha nng c ch vi khun co hi.

Hu ht cc sn phm thng mi (commercial products) ca chng LAB khng th hoc thich nghi yu vi biu m ng rut ca ngi.

 Khi phn tich sn phm dng vin va ht kim tra s lng t bao LAB trn c mi trng BCP va MRS (b sung vi 0,05% Lcysteine) thi khng thy s hin din ca t bao

Nguyn nhn: t bao LAB co th b nh hng bi cc iu kin x l: cc lc nn trong qu trinh dp vin, cc thanh phn ca thuc, va nhit bo qun vv..

 Theo 1 bo co nm 2001, trong 30 ch phm sinh hc dng kh, co 11 ch phm khng c vi khun sng(viable bacteria), ch co cha 7 loi a tuyn b va co hn 18 loai khc so vi nhng loai ghi trn nhn. Kt qu 1 cuc kho st cc loi sn phm Probiotic c bn My cho thy s lng vi khun thp, mt s trng hp khng co, b nhim hoc ghi thng tin sai lch trn nhan. Thiu cc tiu chun(standard) cho cc ch phm Pribiotic