Nghiên cứu đặc tính Probiotic của Bacillus Subtilis BS02

NGHIEN CIAJ DAC TINH PROBIOTIC CUA BACILLUS SUBTILIS BS02

VU THANH THAO, NGUYEN MINH THAI,

NGUYEN TH( LINH GIANG, TRAN HQu TAM, TRAN CAT DONG

Phdng TN Vi Sinh Cdng Ngh§ Dipvc, Khoa O w e, DH Y D irvc TP.HCM

TOM TAT

Trong nghien ciru nay, muc tieu cua chiing tdi la

khao sdt cdc dac tinh probiotic cua chung Bacillus

subtilis BS02 phan lap tir phan ngurii Cdc ddc tinh

probiotic duxyc khao sat theo huring dan cua

WHO/FAO bao gom: kha ndng sinh enzym ngoai

bao, kha ndng doi khdng vdi cac vi khuan gay benh 6

during ruot, kha ndng chiu acid, muoi mat, va cdc yeu

to an toan. Sau dd, kha nang de dieu tri tieu chay

trong mot md hinh chuot BS02 bao tir da duxyc

nghien ciru. Ket qua cho thay BS02 cd the sinh cac

enzym ngoai bdo, ton tai trong dieu kien khde nghiet

cua during tieu hda. Cdc thir nghiem khdng khdng

sinh cho thay BS02 nhay cam vdi 13 khdng sinh

duxyc thir nghiem, ngoai trir ceftazidime. Cuoi cung,

ket qua thir nghiem cho thay bao tir BS02 cd kha

nang dieu tri benh tieu chay tren md hinh chuot khi so

sdnh vdi nhdm doi chirng khdng sir dung probiotic.

Tir khoa: Bacillus subtilis BS02. ddc tinh

probiotic.

SUMMARY

Our objective is studied the probiotic

characteristics of Bacillus subtilis BS02 isolated from

human faeces. Probiotic characteristics were tested

according to the guidelines of WHO/ FAO, which

include: producing extracellular enzymes, competitive

ability against pathogenic bacteria in gastrointestinal

tract, tolerance with acidity in the stomach and bile

salt, and safety aspects. After that, the ability to treat

diarrhea in a mouse model of BS02 spores was

investigated. The results showed that BS02 can

produce extracellular enzymes, survive in the strict

conditions of gastrointestinal tract. The antibiotic

resistance test results showed that BS02 sensitive to

13 tested antibiotics, except ceftazidime. Ultimately,

BS02 spores had capable of treating diarrhea in

mouse model when compared with control group not

used probiotic.

Keywords: Bacillus, probiotic.

OAT VAN 0£

Probiotic la cac che phdm bo sung co chira vi sinh

vat song nhdm tang cudng sire khoe cho ngudi sir

dung bdng cach duy tri sy can bdng vd cai thien he vi

khuan dudng ruot [11] Cac vi khuan thudng dirge

sir dung lam probiotic Id cac vi khuan lactic nhu

Lactobacillus spp. va Bifidobacterium spp. Cac vi

khuan nay dd duac chirng minh tinh an toan qua qua

trinh sir dung lau dai [6 , 10] Tuy nhien, do ddy la

chung vi khuan yem khi va dbi hoi dieu kien dinh

dudng ddc biet nen viec nuoi cay gap nhieu kho khdn

vd gid thanh cao. Ben canh do cac vi khuan ndy

khong chiu duac cac dieu kien khde nghiet cua qua

trinh san xuat nhir dong kho. say tao hat. vd c6

tuoi tho khi bao quan ngdn Trudc day Bacillus duoc

xem la vi khuan song trong ddt do do chiing it duoc

xem xet nhu la probiotic theo nghTa phai thuic nhbm

vi khuan dudng ruot, tuy nhien, gan ddy Tdm vd c$ng

su [14] da chirng minh Bacillus cb the duac xem Id

mot thanh phan cita he vi sinh vat dudng ru6t Trong

nghien ciru trudc day. chung toi da tien hdnh phdn

lap cac chiing Bacillus tir phdn ngudi khoe manh,

trong do chung Bacillus subtilis BS02 duac ddnh gid

la chung co kha ndng tao bao tir cao vd sinh trudng

tot De co the irng dung chung vi khuin ndy nhu

nguon cung c ip probiotic, chiing toi tiep tuc nghiSn

ciru cac ddc tinh probiotic cua chung BS02

OOl TU’QNG VA PHUm iG PHAP NGHIEN ClfU

1 . D6 i tuang

Vi khudn thu nghiem: Chung vi khuin Bacillus

subtilis BS02 duac phdn lap tu phdn ngudi tinh

nguyen khoe manh tai TP. Ho Chi Minh do Phbng Thi

nghiem Vi sinh Cong nghe Duac cung cap

Dong vat thin nghiem: Chuot nhdt trdng gi6ng

Swiss albino, gidi tinh due, the trong chudt tir 18-25

gam duac Vien Pasteur Nha Trang cung cap ChuOt

duac nuoi on dinh trong vong 1 tu in trudc khi ti£n

hanh thir nghiem.

Cac chung vi khudn gay benh: Gom cdc chiing

chuan cua ATCC va cdc chiing duac phdn Idp vd giu

tai Bo mon Vi sinh - Ky sinh: Eschenchia coli ATCC

25922, Pseudomonas aeruginosa ATCC 27853,

Proteus vulgaris ATCC 49132, Staphylococcus

aureus ATCC 43300, Staphylococcus aureus ATCC

29213, Streptococcus feacalis ATCC 29213, Sarcina

marcescens ATCC 14756, Shigella dysenteria,

Salmonella typhi.

Cac thuoc thu Lugol, TCA mua cua Merck.

2. Phuang phap

Phuang phap thu» kha nang sinh enzym ngoai

bao

Thir nghiem kha ndng sinh mot so loai enzym

nhu: protease, amylase, lipase Hoat hda Bacillus

subtilis BS02 tren moi trudng TSB (Tryptic Soy Broth)

trong 2-6 gid. Chuan bi huyen phu vi khuln d mdt d6

108 CFU/ml, cay theo vach thing Ien cac moi trudng

TSA (Tryptic Soy Agar) co chat cam irng thich hap. U

37°C trong 24 gid. Doc ket qua bdng cac thuoc thir

Lugol (amylase) [14], thuoc thir tnchloacetic acid

(protease), quan sat cdc vong due xung quanh kh6m

(lipase). Sau khi dinh tinh kha ndng sinh enzym

chiing toi tien hanh dinh luang hoat tinh enzym ngo?i

bao cua BS02 trong m6i trudng bo sung ca ch£t cam

irng tuang irng vdi hoat tinh cua enzym amylase xdc

dinh theo phuang phdp Heinkel, protease xdc dinh

20 Y HOC THUC HANH (907) - SO 3/2014