Sakon Nakhon Rajabhat University Journal

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6767

ผศ.วิศัย พรหมเทพ

การแยกเชื้อแบคทีเรียสรางกรดแลคติค จากผลิตภัณฑอาหารหมัก

เพื่อใชเปนแหลงของโปรไบโอติก

สุวภา ยศตระโคตร

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68

วารสารมหาวิทยาลัยราชภัฏสกลนคร

øĩĚĴęøĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûÿĩøēĜīČėĨĊĈŧĤĩĢĩĚĢĘĨø ĳĕĮĻĤĶāŤĳĒūĐĴĢĜţþùĤþĵĒĚķđĵĤČīø Isolation of Lactic Acid Bacteria from Fermented Food for Probiotic Source

ēğ.ĞīğĨę ĕĚĢĘĳĎĕ ¹ ġįĞėĩ ęğČħĵûČĚ ¹

đĎûĨċęţĤ

ĎŀĩøĩĚĴęøĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûÿĩøĤĩĢĩĚĢĘĨø 6 ČĨĞĤęţĩþ ĵċęĶāŤĤĩĢĩĚ MRS + 1%

CaCO3 ķċŤĳāĮļĤĴđûĎĬĳĚĬęĎĨļþġīļĐ 52 ķĤĵĂĳĜĎ ĵċęğĭøĠĩÿĩøĜĨøĠĊħĚİĒĚţĩþĴĜħġĨĊćĩĐĞīĎęĩ ĳĘĮĻĤĐŀĩķĒğĭøĠĩ ûįĊġĘđĨČīûĞĩĘĳĒūĐĵĒĚķđĵĤČīø ĕđĳāĮļĤĴđûĎĬĳĚĬęČĨĞĤęţĩþĎĬĻĳĢĘĩħġĘČţĤøĩĚĕĨĉĐĩĳĒūĐÿįĜīĐĎĚĬęŧĵĒĚķđĵĤČīøĎĬĻ ĘĬĒĚħġīĎďīėĩĕċĬĎĬĻġįċûĮĤ ķĤĵĂĳĜĎ ĒĚħČİI11 ĵċęġĩĘĩĚčĳÿĚīĄĳČīđĵČķċŤĶĐġėĩĞħĎĬĻĘĬĳøĜĮĤĐļŀĩċĬ 3 ĳĒĤĚŧĳĂĺĐČŧ ĎĐČţĤġėĩĞħĳĒūĐøĚċ pH 2 ĴĜħĘĬûĞĩĘġĩĘĩĚčĶĐøĩĚęĨđęĨļþĳāĮļĤĴđûĎĬĳĚĬęøţĤĵĚûĒĜĩķċŤ ĵċęġĩĘĩĚčęĨđęĨļþøţĤ ĵĚû Aeromonas hydrophila ĴĜħĳĘĮĻĤĐŀĩĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīû ķĤĵĂĳĜĎ ĒĚħČİI11 ķĒÿĨċÿŀĩĴĐøĶĐĚħċĨđ ġøįĜ ĕđĞţĩĳĒūĐ Lactobacillus sp. ĶĐøĩĚğĭøĠĩčĭþēĜùĤþøĩĚĳġĚīĘ Lactobacillus sp. (ĒĚħČİ I11) ĶĐĒĜĩċįøČţĤ ġĘĚĚčėĩĕċŤĩĐøĩĚēĜīČ ĳāţĐ ĐļŀĩĢĐĨøČĨĞĒĜĩĴĜħĐļŀĩĢĐĨøĘĬāĬĞīČ ĕđĞţĩ øĩĚĳġĚīĘĤĩĢĩĚċŤĞę Lactobacillus sp.

(ĒĚħČİ I11) ĎŀĩĶĢŤġĘĚĚčėĩĕċŤĩĐĐļŀĩĢĐĨøČĨĞĒĜĩĴĜħĐļŀĩĢĐĨøĘĬāĬĞīČĳĕīĻĘùĭļĐ ċĨþĐĨļĐĳĕĮĻĤĳĕīĻĘġĘĚĚčėĩĕøĩĚēĜīČ ĒĜĩċįøĶĢŤċĬùĭļĐÿĭþûĞĚĳġĚīĘĤĩĢĩĚċŤĞę Lactobacillus sp. (ĒĚħČİ I11)

ûŀĩġŀĩûĨĄ : ĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīû, ĴĢĜţþĵĒĚķđĵĤČīø

Abstract

Six fermented food samples were collected and used as sources to isolate lactic acid bacteria by using MRS + 1% CaCO3 medium. 52 isolates obtained were found lactic acid bacteria by studying cell morphology and physiology. Study on certain properties of isolates found only one strain (door I11), which

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6969 Sakon Nakhon Rajabhat University Journal

had potential to be used as the probiotic. It was tolerant to 3% bile and showed degrees of survival pH 2 and had antimicrobial activities against Aeromonas hydrophila. Based on morphology and biochemical characteristics analysis, door I11 was identified as Lactobacillus sp.. The effect of Lactobacillus sp. (door I11) on production performance in cat fishs were investigated. The results showed that supplementation of Lactobacillus sp. (door I11) in cat fishs diets improved production performance, especially weight gain and live weight efficiency. For optimum performance of cat fishs, Lactobacillus sp. (door I11) should be added as supplementation in diets.

Keyword : Lactic acid bacteria, Probiotic source

đĎĐŀĩ

ĴđûĎĬĳĚĬęøĚċĴĜûČīû ĢĚĮĤ LAB (Lactic Acid Bacteria) ĘĬûĞĩĘġŀĩûĨĄČţĤøĚħđĞĐøĩĚĢĘĨøĤĩĢĩĚ ĘĬ ĒĚħĵęāĐŧĶĐċŤĩĐġţþĳġĚīĘġįùėĩĕùĤþĘĐįĠęŧĴĜħġĨČĞŧĢĚĮĤĘĬûįĊġĘđĨČīĳĒūĐ Probiotic ĳāţĐ ġĩęĕĨĐďįŧĳĀĕĩħùĤþĳāĮļĤ Lb. acidophilus, Lb. casei ĴĜħ Lb. rhamnosus ġĩĘĩĚčĚĨøĠĩûĞĩĘēīċĒøČīĶĐøĩĚķĘţġĩĘĩĚčęţĤęĐļŀĩČĩĜ ĴĜĺûĵČġ ĢĚĮĤøĩĚČīċĳāĮļĤĴđûĎĬĳĚĬęøţĤĵĚûĎĩþĳċīĐĤĩĢĩĚĎĬĻĎŀĩĶĢŤĳøīċĤĩøĩĚĤįÿÿĩĚħĚţĞþĶĐĳċĺøĎĩĚøĴĜħĤĩøĩĚ ĚħđđĎĩþĳċīĐĤĩĢĩĚĤĨøĳġđĢĚĮĤĤĩøĩĚĤįÿÿĩĚħĚţĞþĶĐēİŤĶĢĄţĳĐĮĻĤþÿĩøøĩĚĶāŤęĩĒĆīāĬĞĐħ ĐĤøÿĩøĐĬļęĨþĕđĞţĩ Lb.acidophilus ĘĬēĜČŤĩĐøĩĚĳøīċėĩĞħøĩĚĘĬûĜĤĳĚġĳČĤĚĤĜġİþ (hypocholesterol) ĴĜħęĨþĘĬĚĩęþĩĐøĩĚĞīÿĨęĞţĩ ĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûđĩþġĩęĕĨĐďįŧĘĬēĜĳĒūĐġĩĚČŤĩĐøţĤĘħĳĚĺþ (antimutagenic) ġĩĚČŤĩĐĳĐĮļĤþĤø (antitumor) ĢĚĮĤøĚħČįŤĐėİĘīûįŤĘøĨĐĶĐĘĐįĠęŧ ĴĜħēĜĳāţĐĳċĬęĞøĨĐĐĬļøĺĳøīċùĭļĐĶĐġĨČĞŧ ÿĩøûįĊġĘđĨČīùŤĩþČŤĐùĤþ ĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīû øĩĚĶāŤġĩĚĳġĚīĘāĬĞĐħ (Probiotics) ĶĐĤĩĢĩĚġĨČĞŧĳĒūĐĴĐĞĎĩþĢĐĭĻþġŀĩĢĚĨđøĩĚ ĕĨĉĐĩĤįČġĩĢøĚĚĘĤĩĢĩĚġĨČĞŧ ĳĕĚĩħĶĐĒũÿÿįđĨĐøĩĚĶāŤęĩĒĆīāĬĞĐħēġĘĶĐĤĩĢĩĚġĨČĞŧĐĨļĐøţĤĶĢŤĳøīċĒũĄĢĩĘĩø ĳāţĐ øĩĚċĮļĤęĩùĤþġĨČĞŧ ġĩĚČøûŤĩþĶĐĳĐĮļĤġĨČĞŧ ċĨþĐĨļĐøĩĚĳĜĮĤøĶāŤġĩĚĳġĚīĘāĬĞĐħĶĐĤĩĢĩĚġĨČĞŧĐţĩÿħĳĒūĐĞīďĬĎĬĻċĬ ĶĐċŤĩĐøĩĚĶāŤĒĚħĵęāĐŧ ġţþĳġĚīĘġįùėĩĕùĤþĘĐįĠęŧĴĜħġĨČĞŧĎĬĻĒĜĤċėĨę ĵċęþĩĐĞīÿĨęĐĬļĘĬÿįċĒĚħġþûŧ ûĮĤĴęøĳāĮļĤ ĴđûĎĬĳĚĬęĎĬĻġĚŤĩþøĚċĴĜûČīûÿĩøēĜīČėĨĊĈŧĤĩĢĩĚĢĘĨøĳĕĮĻĤĶāŤĳĒūĐĴĢĜţþùĤþ Probiotic

ĞĨČčįĒĚħġþûŧùĤþøĩĚĞīÿĨę

1. ĴęøĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûÿĩøēĜīČėĨĊĈŧĤĩĢĩĚĢĘĨøĳĕĮĻĤĶāŤĳĒūĐĴĢĜţþùĤþÿįĜīĐĎĚĬęŧ Probiotic

2. ğĭøĠĩûįĊġĘđĨČīđĩþĒĚħøĩĚùĤþÿįĜīĐĎĚĬęŧĵĒĚķđĵĤČīøùĤþĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĬĻĴęøķċŤ 3. ğĭøĠĩĤĐįøĚĘĞīďĩĐùĤþĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜøČīûĎĬĻĘĬûįĊġĘđĨČī Probiotic

4. ĕĨĉĐĩûįĊėĩĕĤĩĢĩĚĒĜĩĶĢŤĘĬûįĊėĩĕĳĕīĻĘĘĩøùĭļĐÿĩøÿįĜīĐĎĚĬęŧ Probiotic

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70

ĞīďĬċŀĩĳĐīĐøĩĚĞīÿĨę

1. øĩĚĴęøĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûÿĩøēĜīČėĨĊĈŧĤĩĢĩĚĢĘĨøĳĕĮĻĤĶāŤĳĒūĐĴĢĜţþùĤþÿįĜīĐĎĚĬęŧ Probiotic ĐŀĩČĨĞĤęţĩþĜħ 25 øĚĨĘ ČĬĒŞĐĶĐġĩĚĜħĜĩęĵĂĳċĬęĘûĜĤķĚġŧ 0.85 ĳĒĤĚŧĳĂĺĐČŧ ĒĚīĘĩČĚ 225 ĘīĜĜīĜīČĚ ĵċęĳûĚĮĻĤþČĬĒŞĐĤĩĢĩĚ ĎŀĩøĩĚĳÿĮĤÿĩþČĨĞĤęţĩþ ĴĜŤĞĳĕĩħĳāĮļĤċŤĞęĞīďĬøĩĚ spread plate ĵċęĶāŤĤĩĢĩĚĳĕĩħĳāĮļĤ Ĵùĺþ MRS ĎĬĻĳČīĘĴûĜĳĂĬęĘûĩĚŧđĤĳĐČ 1 ĳĒĤĚŧĳĂĺĐČŧ đţĘĎĬĻĤįĊĢėİĘī 37 ĤþğĩĳĂĜĳĂĬęġ ĳĒūĐĳĞĜĩ 24 – 48 āĨĻĞĵĘþ ĶĐġėĩĞħķĚŤĤĤøĂīĳÿĐ ÿĩøĐĨļĐĳĜĮĤøĵûĵĜĐĬĎĬĻĘĬđĚīĳĞĊĶġĚĤđĵûĵĜĐĬ ĎŀĩøĩĚùĬċĳāĮļĤ (cross streak) đĐĤĩĢĩĚ ĳĕĩħĳāĮļĤĴùĺþ MRS Ăļŀĩ 2 ûĚĨļþ ĳĕĮĻĤĶĢŤķċŤĳāĮļĤđĚīġįĎďīĿ ĴĜħČĚĞÿġĤđûĞĩĘđĚīġįĎďīĿĵċęġţĤþċİċŤĞęøĜŤĤþÿįĜĎĚĚğĐŧ

2. ğĭøĠĩûįĊġĘđĨČīđĩþĒĚħøĩĚùĤþÿįĜīĐĎĚĬęŧĵĒĚķđĵĤČīøùĤþĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĬĻĴęøķċŤ 2.1 øĩĚĎċġĤđûĞĩĘġĩĘĩĚčĎĐČţĤĐļŀĩċĬùĤþĴđûĎĬĳĚĬęøĚċĴĜûČīø ċĨċĴĒĜþÿĩøĞīďĬùĤþ Jin et al.

(1998) ĵċęčţĩęĳāĮļĤ 1 loop ĜþĶĐĤĩĢĩĚĳĢĜĞ MRS ĒĚīĘĩČĚ 5 ĘīĜĜīĜīČĚ ĐŀĩķĒđţĘĎĬĻĤįĊĢėİĘī 37 Ĥþğĩ ĳĂĜĳĂĬęġĐĩĐ 18-24 āĨĻĞĵĘþ ÿĩøĐĨļĐĐŀĩĘĩĳùĬĻęđĐĤĩĢĩĚĴùĺþ MRS đţĘĎĬĻĤįĊĢėİĘī 37 ĤþğĩĳĂĜĳĂĬęġĐĩĐ 18-24 āĨĻĞĵĘþ čţĩęĳāĮļĤ 1 ĵûĵĜĐĬĜþĶĐĤĩĢĩĚĳĢĜĞ MRS ĒĚīĘĩČĚ 5 ĘīĜĜīĜīČĚ đţĘĎĬĻĤįĊĢėİĘī 37 ĤþğĩĳĂĜĳĂĬęġ ĐĩĐ 18-24 āĨĻĞĵĘþĴĜŤĞčţĩęĳāĮļĤ 50 ķĘĵûĚĜīČĚĜþĶĐĤĩĢĩĚĳĢĜĞ MRS 5 ĘīĜĜīĜīČĚĎĬĻĘĬĐļŀĩċĬķøţġċ 3 ĳĒĤĚŧĳĂĺĐČŧ (ĐļŀĩĢĐĨøČţĤĒĚīĘĩČĚ) ĴĜħĤĩĢĩĚĳĢĜĞ MRS ĎĬĻķĘţĘĬĐļŀĩċĬ 5 ĘīĜĜīĜīČĚĳĒūĐāįċûĞđûįĘ đţĘĎĨļþ 2 āįċĎċĜĤþĎĬĻ ĤįĊĢėİĘī 37 ĤþğĩĳĂĜĳĂĬęġĐĩĐ 2 āĨĻĞĵĘþ ĢĜĨþÿĩøĐĨļĐĞĨċĒĚīĘĩĊĳāĮļĤĎĬĻĘĬāĬĞīČċŤĞęĞīďĬøĩĚ spread plate ĐŀĩēĜĎĬĻ ķċŤĘĩûŀĩĐĞĊĢĩûţĩøīÿøĚĚĘøĩĚĎĐČţĤĐļŀĩċĬ

2.2 øĩĚĎċġĤđûĞĩĘġĩĘĩĚčĶĐøĩĚĎĐČţĤøĚċĴĜħĳđġ ċĨċĴĒĜþĘĩÿĩøĞīďĬùĤþ Jin et al. (1998) ĵċęčţĩęĳāĮļĤ 1 loop ÿĩø stock culture ĜþĶĐĤĩĢĩĚĳĢĜĞ MRS ĒĚīĘĩČĚ 5 ĘīĜĜīĜīČĚ ĐŀĩķĒđţĘĎĬĻĤįĊĢėİĘī 37 ĤþğĩĳĂĜĳĂĬęġĐĩĐ 18-24 āĨĻĞĵĘþ ÿĩøĐĨļĐĐŀĩĘĩĳùĬĻęđĐĤĩĢĩĚĴùĺþ MRS đţĘĎĬĻĤįĊĢėİĘī 37 ĤþğĩĳĂĜĳĂĬęġĐĩĐ 18-24 āĨĻĞĵĘþ čţĩęĳāĮļĤđĚīġįĎďīĿ 1 ĵûĵĜĐĬĶĐĤĩĢĩĚĳĢĜĞ MRS ĒĚīĘĩČĚ 5 ĘīĜĜīĜīČĚ ĴĜħđţĘĎĬĻĤįĊĢėİĘī 37 Ĥþğĩ ĳĂĜĳĂĬęġĐĩĐ 18-24 āĨĻĞĵĘþ čţĩęĳāĮļĤ 50 ķĘĵûĚĜīČĚĜþĶĐĤĩĢĩĚĳĢĜĞ MRS 5 ĘīĜĜīĜīČĚ ĎĬĻĒĚĨđ pH 2 ĵċęĶāŤ hydrochloric acid ûĞĩĘĳùŤĘùŤĐ 1 ĵĘĜĩĚŧ ĶĐøĩĚĒĚĨđ MRS ĶĢŤĳĒūĐøĚċ ĞīĳûĚĩħĢŧĒĚīĘĩĊĳāĮļĤĳĚīĻĘČŤĐĎĬĻ 0 āĨĻĞĵĘþ ĴĜħĢĜĨþÿĩøđţĘĎĬĻĤįĊĢėİĘī 37 ĤþğĩĳĂĜĳĂĬęġĐĩĐ 3 āĨĻĞĵĘþ ĵċęĞīďĬøĩĚ spread plate ĴġċþēĜĳĒūĐûţĩ ĳĒĤĚŧĳĂĺĐČŧøĩĚĤęİţĚĤċ

2.3 øĩĚĎċġĤđûĞĩĘġĩĘĩĚčęĨđęĨļþĳāĮļĤøţĤĵĚû ĵċęĞīďĬ Paper - disc diffusion ĐŀĩķĘŤĕĨĐġŀĩĜĬýţĩ ĳāĮļĤÿįţĘġĩĚĜħĜĩęĳāĮļĤĎċġĤđĎĬĻĳČĚĬęĘķĞŤ swab ĜþđĐĤĩĢĩĚĳĜĬļęþĳĕĩħĳāĮļĤĴùĺþ NA + 1 % NaCl ĴĜħĤĩĢĩĚĴùĺþ MRS ČĩĘĜŀĩċĨđ ÿĩøĐĨļĐĐŀĩøĚħċĩĠøĚĤþùĐĩċĳġŤĐēţĩğİĐęŧøĜĩþ 5 ĘīĜĜīĳĘČĚ ĎĬĻýţĩĳāĮļĤĞĩþĜþĢĐŤĩĤĩĢĩĚĴùĺþ ĎĨļþġĤþ ĴĜŤĞĢęċĐļŀĩĳĜĬļęþĳāĮļĤĒĚĩğÿĩøĳĂĜùĤþĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĬĻēţĩĐøĩĚĒĚĨđ pH ĶĢŤĳĒūĐøĜĩþ (pH 7) ĒĚīĘĩĊ 20 ķĘĵûĚĜīČĚĜþķĒ ĐŀĩķĒđţĘĎĬĻĤįĊĢėİĘī 37 ĤþğĩĳĂĜĳĂĬęġ ĳĒūĐĳĞĜĩ 24 āĨĻĞĵĘþ ĴĜŤĞČĚĞÿġĤđøĩĚ ęĨđęĨļþøĩĚĳÿĚīĄùĤþĳāĮļĤĎċġĤđĵċęûŀĩĐĞĊĒĚħġīĎďīėĩĕøĩĚęĨđęĨļþĳĒūĐøīÿøĚĚĘøĩĚęĨđęĨļþ (inhibition activity)

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3. øĩĚÿĨċÿŀĩĴĐøĶĐĚħċĨđÿĬĐĨġ ĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĬĻġĚŤĩþĴđûĎĬĚīĵĤĂīĐķċŤ ċĨċĴĒĜþĘĩÿĩøĞīďĬ Beisheier et al. (1991)

3.1 øĩĚČīċġĬĴøĚĘ ĚİĒĚţĩþĴĜħøĩĚġĚŤĩþøťĩĂĳĘĮĻĤĳÿĚīĄĶĐĤĩĢĩĚ MRS broth 3.2 øĩĚĳÿĚīĄĶĐĤĩĢĩĚ MRS broth ĎĬĻĤįĊĢėİĘī 10 ĴĜħ 45°C

3.3 øĩĚĳÿĚīĄĶĐĤĩĢĩĚ MRS broth ĎĬĻĘĬĵĂĳċĬęĘûĜĤķĚċŧ 6.5 ĴĜħ 18 % 3.4 øĩĚĳÿĚīĄĶĐĤĩĢĩĚ MRS broth ĎĬĻĒĚĨđûţĩ pH ĳĒūĐ 4.4 ĴĜħ 9.6

4. øĩĚĕĨĉĐĩûįĊėĩĕĤĩĢĩĚĒĜĩĳĕĮĻĤĶĢŤĒĜĩĘĬûįĊėĩĕĳĕīĻĘĘĩøùĭļĐÿĩøÿįĜīĐĎĚĬęŧ Probiotic ĶĐøĩĚ ĎċĜĤþķċŤĞĩþĴēĐøĩĚĎċĜĤþĴđđġįţĘČĜĤċ ĂĭĻþĴđţþøĩĚĎċĜĤþĳĒūĐ 2 øĜįţĘČĩĘġİČĚĤĩĢĩĚ ĵċęĶāŤĜİøĒĜĩċįø ĤĩęįĳĚīĻĘĎċĜĤþ 1 ĳċĮĤĐ øĜįţĘøĩĚĎċĜĤþĜħ 15 ČĨĞ ĚĞĘĎĨļþĢĘċ 30 ČĨĞ ĎŀĩøĩĚĎċĜĤþĳĒūĐĚħęħĳĞĜĩ 40 ĞĨĐ ĂĭĻþ ĘĬøĩĚĴđţþøĜįţĘøĩĚĎċĜĤþ ċĨþĐĬļ

øĜįţĘĎĬĻ 1 ĶĢŤĤĩĢĩĚĕĮļĐćĩĐķĘţĳġĚīĘĵĒĚķđĵĤČīø

øĜįţĘĎĬĻ 2 ĶĢŤĤĩĢĩĚĕĮļĐćĩĐĳġĚīĘĵĒĚķđĵĤČīøĚĢĨġĒĚħČİ l11

ĎŀĩøĩĚĳĜĬļęþĒĜĩċįøĳĒūĐĳĞĜĩ 20 ĞĨĐ ÿĩøĐĨļĐĶġţĳāĮļĤĴđûĎĬĳĚĬęøţĤĵĚûĶĐĒĚīĘĩĊĳāĮļĤĜħ 10⁸CFU/ml ĜþĶĐđţĤĳĜĬļęþĒĜĩĎĨļþ 2 øĜįţĘ ÿĩøĐĨļĐĳĜĬļęþĒĜĩÿĐûĚđ 40 ĞĨĐ ÿĩøĐĨļĐđĨĐĎĭøĴĜħĞīĳûĚĩħĢŧùŤĤĘİĜ

ēĜøĩĚĞīÿĨę

1. ēĜøĩĚĴęøĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûÿĩøēĜīČėĨĊĈŧĤĩĢĩĚĢĘĨøĳĕĮĻĤĶāŤĳĒūĐĴĢĜţþùĤþ ÿįĜīĐĎĚĬęŧ Probiotic

ēĜøĩĚûĨċĴęøĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûÿĩøēĜīČėĨĊĈŧĤĩĢĩĚĢĘĨøČţĩþĹ ûĮĤ ĒĜĩĚŤĩ ĴĢĐĘøįŤþ ÿţĤĘ ġŤĘĳĎŤĩĞĨĞ ĴĜħĢĐţĤķĘŤċĤþ ċŤĞęĳĎûĐīû spread plate đĐĤĩĢĩĚ MRS ĎĬĻĳČīĘ CaCo3 ĚŤĤęĜħ 1 ĐŀĩÿĩĐ ĤĩĢĩĚĳĜĬļęþĳāĮļĤķĒđţĘĎĬĻĤįĊĢėİĘī 37 ĤþğĩĳĂĜĳĂĬęġĳĒūĐĳĞĜĩ 24 āĨĻĞĵĘþ ĶĐėĩĞħķĚŤĤĤøĂīĳÿĐ (anaerobic condition) ÿĩøĐĨļĐĳĜĮĤøĴđûĎĬĳĚĬęĎĬĻĘĬđĚīĳĞĊĶġĚĤđĵûĵĜĐĬķĒĎŀĩĶĢŤđĚīġįĎďīĿċŤĞęĳĎûĐīû cross streak đĐĤĩĢĩĚ MRS ÿĩøøĩĚĎċĜĤþĕđĞţĩġĩĘĩĚčûĨċĴęøĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûķċŤĎĨļþĢĘċ 52 ķĤĵĂĳĜČ

2. ēĜøĩĚğĭøĠĩûįĊġĘđĨČīđĩþĒĚħøĩĚùĤþÿįĜīĐĎĚĬęŧĵĒĚķđĵĤČīøùĤþĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċ ĴĜûČīûĎĬĻĴęøķċŤ

2.1 ēĜøĩĚĎċġĤđûĞĩĘġĩĘĩĚčĎĐČţĤĐļŀĩċĬùĤþĴđûĎĬĳĚĬęøĚċĴĜøČīø øĩĚĴęøĴđûĎĬĳĚĬęġĚŤĩþ øĚċĴĜûČīû ÿŀĩĐĞĐ 52 ķĤĵĂĳĜČ ĎĬĻĘĬûįĊġĘđĨČīĵĒĚķđĵĤČīøĶĐĳđĮļĤþČŤĐĳĒūĐøĩĚûĨċĳĜĮĤøÿĩøûĞĩĘġĩĘĩĚčĶĐ øĩĚĎĐČţĤĐļŀĩċĬ ĳĐĮĻĤþÿĩøĐļŀĩċĬĳĒūĐġĩĚāţĞęęţĤęġĩĚĤĩĢĩĚĒĚħĳėĎķùĘĨĐĶĐĜŀĩķġŤĳĜĺøùĤþġĨČĞŧ ĘĬûįĊġĘđĨČīĎŀĩĶĢŤ ķùĘĨĐĴČøČĨĞęţĤęġĩĚĤĩĢĩĚĒĚħĳėĎķùĘĨĐĶĐĜŀĩķġŤĳĜĺøùĤþġĨČĞŧ (Eugene et al., 1996) ÿĭþġĩĘĩĚčĎŀĩĜĩęĳęĮĻĤ ĢįŤĘĳĂĜùĤþĴđûĎĬĳĚĬęĎŀĩĶĢŤĴđûĎĬĳĚĬęČĩęķċŤ (Jin et al., 1998) ċĨþĐĨļĐ ûĞĩĘġĩĘĩĚčĶĐøĩĚĎĐČţĤĐļŀĩċĬÿĭþĳĒūĐ

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72

ĒũÿÿĨęġŀĩûĨĄĶĐøĩĚûĨċĳĜĮĤøĳāĮļĤĎĬĻÿħĐŀĩķĒēĜīČĵĒĚķđĵĤČīø ÿĩøøĩĚĎċġĤđûĞĩĘġĩĘĩĚčĎĐČţĤĐļŀĩċĬùĤþ ĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûÿŀĩĐĞĐ 52 ķĤĵĂĳĜČ ĎĬĻĚħċĨđûĞĩĘĳùŤĘùŤĐùĤþĐļŀĩċĬĳĎţĩøĨđ 3 ĳĒĤĚŧĳĂĺĐČŧ ĳĒūĐĳĞĜĩ 2 āĨĻĞĵĘþ ĕđĞţĩ ĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīû 26 ķĤĵĂĳĜČ ĘĬûĞĩĘġĩĘĩĚčĎĐČţĤĐļŀĩċĬķċŤ ķċŤĴøţ ķĤĵĂĳĜČ ĒĜĩĚŤĩ I3, ĒĜĩĚŤĩI4, ĒĜĩĚŤĩI5, ĒĜĩĚŤĩ10, ġŤĘĳĎŤĩĞĨĞI1, ġŤĘĳĎŤĩĞĨĞI3, øįŤþI1, øįŤþI5, øįŤþI7, øøI1, øøI2, ĞĚ1, ĞĚ2, ĞĚ4, ĞĚ5,ĞĚ6,ĞĚ7,ĞĚ8, ĒĚħČİI2, ĒĚħČİI4, ĒĚħČİI9, ĒĚħČİI11, ĒĚħČİI12, ĒĚħČİI 14, ĒĚħČİI 15ĴĜħĒĚħČİI 16 ĂĭĻþĳāĮļĤ ĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĨļþ 26 ķĤĵĂĳĜČĐţĩÿħĘĬĒĚħĵęāĐŧČţĤøĩĚĚĨøĠĩġĘċįĜùĤþÿįĜīĐĎĚĬęŧĶĐĚħđđĎĩþĳċīĐ ĤĩĢĩĚķċŤ ĳĐĮĻĤþÿĩøÿįĜīĐĎĚĬęŧĎĨļþ 26 ķĤĵĂĳĜČ ġĩĘĩĚčĎĐČţĤġėĩĞħøċċĨĐĎĬĻĘĬĳøĜĮĤĐļŀĩċĬĶĐĜŀĩķġŤĳĜĺøĎĬĻčİøġţþĘĩ ÿĩøčįþĐļŀĩċĬĶĐČĨđĶĐĚİĒùĤþ conjugate bile acid ĳøĜĮĤĐļŀĩċĬÿħčİøĢĘįĐĳĞĬęĐ 10 – 12 ûĚĨļþČţĤĞĨĐ ĳĕĮĻĤĎŀĩĒąīøīĚīęĩ øĨđĤĩĢĩĚĒĚħĳėĎķùĘĨĐĴĜħĵûĳĜġĳČĤĚĤĜ ĢĜĨþÿĩøĐĨļĐÿħčİøċİċĂĭĘēţĩĐĳġŤĐĳĜĮĤċċŀĩķĒęĨþČĨđĳĕĮĻĤĴĒĚĚİĒĴĜħ ĢĘįĐĳĞĬęĐĐŀĩøĜĨđĘĩĶĢĘţĂĭĻþĳĚĬęøĞţĩ deconjugated bile acid ċĨþĐĨļĐøĩĚĳøīċøĚħđĞĐøĩĚċĨþøĜţĩĞÿĭþĘĬđĎđĩĎ ġŀĩûĨĄĶĐøĩĚĚĨøĠĩġĘċįĜùĤþÿįĜīĐĎĚĬęŧĶĐĚħđđĜŀĩķġŤ ĴĜħøĩĚğĭøĠĩûįĊġĘđĨČīøĩĚĎĐČţĤĐļŀĩċĬ ĴġċþĶĢŤĳĢĺĐĞţĩ ĴđûĎĬĳĚĬęġĩĘĩĚčĎĬĻÿħĎĐĤęİţķċŤĶĐĜŀĩķġŤ ĳĐĮĻĤþÿĩøĐļŀĩċĬĘĬûįĊġĘđĨČīĶĐøĩĚĎŀĩĜĩęĴđûĎĬĳĚĬęķċŤ ĐĤøÿĩøĐĨļĐĐļŀĩċĬ ęĨþĘĬěĎďīĿĶĐøĩĚęĨđęĨļþĴđûĎĬĳĚĬę ĵċęøĩĚęţĤęġĜĩęķùĘĨĐĴĜħøĚċķùĘĨĐČţĩþĹ ĎĬĻĳĒūĐġţĞĐĒĚħøĤđùĤþēĐĨþĳĂĜ ĳøīċĳĒūĐāţĤþĞţĩþĎŀĩĶĢŤûĞĩĘġĩĘĩĚčĶĐøĩĚûĞđûįĘøĩĚēţĩĐĳùŤĩĤĤøùĤþġĩĚ (permeability) ġİĄĢĩęķĒ (Jin et al. 1998)

2.2 ēĜøĩĚĎċġĤđûĞĩĘġĩĘĩĚčĶĐøĩĚĎĐČţĤøĚċ ûĞĩĘġĩĘĩĚčĶĐøĩĚĎĐČţĤøĚċĳĒūĐûįĊġĘđĨČī ùĤþĴđûĎĬĳĚĬęĎĬĻĘĬûįĊġĘđĨČīĳĒūĐĵĒĚķđĵĤČīø (Havenaar et al. 1992) ĳĐĮĻĤþÿĩøĶĐĚħđđĎĩþĳċīĐĤĩĢĩĚùĤþûĐ ĴĜħġĨČĞŧ ĘĬøĩĚĢĜĨĻþøĚċĳøĜĮĤĎŀĩĶĢŤ pH ĶĐĚħđđĎĩþĳċīĐĤĩĢĩĚĜċĜþĳĎţĩøĨđ 0.5 – 2.0 ġţþēĜČţĤĴđûĎĬĳĚĬęġţĞĐ ĶĢĄţĎŀĩĶĢŤķĘţġĩĘĩĚčĎĐĴĜħĳÿĚīĄķċŤ (Jin et al. 1998) ċĨþĐĨļĐ ĶĐøĩĚğĭøĠĩĐĬļÿĭþķċŤĐŀĩĴđûĎĬĳĚĬęġĚŤĩþøĚċ ĴĜûČīûĎĬĻġĩĘĩĚčĎĐČţĤĐļŀĩċĬ ÿŀĩĐĞĐ 26 ķĤĵĂĳĜČ ĘĩûĨċĳĜĮĤøĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĬĻġĩĘĩĚčĎĐČţĤøĚċ ĳĕĮĻĤġĩĘĩĚčĳÿĚīĄĴĜħĤęİţĚĤċķċŤĶĐĚħđđĎĩþĳċīĐĤĩĢĩĚùĤþġĨČĞŧĳÿŤĩčīĻĐ ĴĜħÿĩøøĩĚĎċĜĤþĕđĞţĩ ĳāĮļĤ ĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûķĤĵĂĳĜČĎĬĻĎĐČţĤ pH 2 ĘĬĎĨļþĢĘċ 10 ķĤĵĂĳĜČûĮĤ ĞĚI5, ĒĜĩĚŤĩI5, ġŤĘĳĎŤĩĞĨĞI1, øįŤþ I1, øøI2, ĒĚħČİI4, ĒĚħČİI9, ĒĚħČİI11, ĒĚħČİI12 ĴĜħĒĚħČİI16

2.3 ēĜøĩĚĎċġĤđûĞĩĘġĩĘĩĚčęĨđęĨļþĳāĮļĤøţĤĵĚû øĩĚğĭøĠĩûĞĩĘġĩĘĩĚčęĨđęĨļþĳāĮļĤøţĤĵĚû ĳĐĮĻĤþÿĩø ĴđûĎĬĳĚĬęĵĒĚķđĵĤČīøĘĬûĞĩĘġĩĘĩĚčĶĐøĩĚġţþĳġĚīĘĶĢŤġĨČĞŧĳÿŤĩđŤĩĐĘĬġįùėĩĕĎĬĻċĬ øĩĚęĨđęĨļþĳāĮļĤøţĤ ĵĚûÿĭþĳĒūĐĒũÿÿĨęĢĐŤĩĎĬĻġŀĩûĨĄĶĐøĩĚûĨċĳĜĮĤøĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīû ĎŀĩĵċęĐŀĩĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜ ûČīûĎĬĻēţĩĐûįĊġĘđĨČīøĩĚĳÿĚīĄĳČīđĵČķċŤĶĐøĚċ pH 2 ÿŀĩĐĞĐ 10 ķĤĵĂĳĜČ ĘĩğĭøĠĩĒĚħġīĎďīėĩĕøĩĚęĨđęĨļþ ÿįĜīĐĎĚĬęŧĎĬĻĘĨøĒūĐġĩĳĢČįøĩĚĳøīċĵĚûĶĐĒĜĩ 2 āĐīċ ûĮĤ Aeromonas hydrophila ĴĜħ Streptococcus sp. ĵċęĞīďĬ paper - disc diffusion čŤĩĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûġĩĘĩĚčēĜīČġĩĚęĨđęĨļþĳāĮļĤÿįĜīĐĎĚĬęŧøţĤĵĚûķċŤÿħĕđ đĚīĳĞĊĶġĳøīċĶĐĚĤđĴēţĐ disc ĎċġĤđĵċęĐŀĩĐļŀĩĳĜĬļęþĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĘĩĎċġĤđ ĕđĞţĩ ĘĬĳĕĬęþ

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7 ķĤĵĂĳĜČĳĎţĩĐĨļĐĎĬĻġĩĘĩĚčęĨđęĨļþĳāĮļĤøţĤĵĚû Aeromonas hydrophila ķċŤûĮĤ ĒĜĩĚŤĩI5, ġŤĘČĬĐĞĨĞI1, ĒĚħČİI4, ĒĚħČİI9, ĒĚħČİI11, ĒĚħČİI12, ĴĜħĒĚħČİI16 ĵċęķĤĵĂĳĜČĎĬĻęĨđęĨļþøĩĚĳÿĚīĄķċŤċĬĎĬĻġįċ ûĮĤ ĒĚħČİI11 ĘĬùĐĩċ ĳġŤĐēţĩğİĐęŧøĜĩþĳĎţĩøĨđ 1 ĳĂĐČīĳĘČĚ ÿĩøēĜøĩĚĎċĜĤþęĨþĕđĤĬøĞţĩĜĨøĠĊħùĤþđĚīĳĞĊĶġĎĬĻĳøīċùĭļĐĚĤđ disc ęĨþĶġĴĜħāĨċĳÿĐøĞţĩķĤĵĂĳĜČĤĮĻĐĹ ċĨþĴġċþĶĐėĩĕĎĬĻ 1

ėĩĕĎĬĻ 1 đĚīĳĞĊĶġĎĬĻĳøīċÿĩøøĩĚęĨđęĨļþĳāĮļĤĴđûĎĬĳĚĬęøţĤĵĚû Aeromonas hydrophila ùĤþĐļŀĩĳĜĬļęþĳāĮļĤ ĴđûĎĬĳĚĬęġĚŤĩþ øĚċĴĜûČīû ķĤĵĂĳĜČĒĚħČİ I11

3. ēĜøĩĚğĭøĠĩĤĐįøĚĘĞīďĩĐùĤþĴđûĎĬĳĚĬęøĚċĴĜøČīøĎĬĻĘĬûįĊġĘđĨČī probiotic

ĐŀĩĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûķĤĵĂĳĜČĒĚħČİ I11 ğĭøĠĩûįĊġĘđĨČīĎĩþġĨĊćĩĐĞīĎęĩĴĜħĎĩþāĬĞĳûĘĬ ĵċęĐŀĩùŤĤĘİĜĎĬĻķċŤĘĩĳĎĬęđĳûĬęþøĨđùŤĤĘİĜ Bergey 's Manual of Systemic Bacteriology (Sneath. 1986) ĞīďĬøĩĚ ùĤþĐīċĩ ĤĩđġįĞĚĚĊŧ (2550 ĤŤĩþčĭþ Salmiminen Wright.1993) ĕđĞţĩĳāĮļĤķĤĵĂĳĜČĒĚħČİ I11 ĳĒūĐĴđûĎĬĳĚĬę ĴøĚĘđĞø ĚİĒĚţĩþĳĒūĐĎţĤĐġĨļĐ ķĘţġĚŤĩþĳĤĐķĂĘŧûħČħĳĜġ ġĚŤĩþøĚċÿĩøøĩĚĢĘĨøĐļŀĩČĩĜøĜİĵûġ ĳĒūĐĴđûĎĬĳĚĬęĎĬĻ ĢĘĨøĴđđ Homofermentative ķĘţĳÿĚīĄĶĐĤĩĢĩĚĎĬĻĘĬĳøĜĮĤĚŤĤęĜħ 6.5 ĴĜħ 18 ĳÿĚīĄķċŤĎĬĻ 10 ĴĜħ 45 Ĥþğĩ ĳĂĜĳĂĬęġ ĳÿĚīĄķċŤĎĬĻ pH 4.4 ĴČţķĘţġĩĘĩĚčĳÿĚīĄķċŤĎĬĻ pH 9.6 ĴĜħķĘţġĩĘĩĚčĳûĜĮĻĤĐĎĬĻķċŤ ĕđĞţĩ ĳĒūĐĴđûĎĬĳĚĬę ġĚŤĩþøĚċĴĜûČīûĶĐġøįĜ Lactobacillus sp.

4. øĩĚĕĨĉĐĩûįĊėĩĕĤĩĢĩĚĒĜĩĶĢŤĒĜĩĘĬûįĊėĩĕĳĕīĻĘĘĩøùĭļĐÿĩøÿįĜīĐĎĚĬęŧ Probiotic

ĶĐøĩĚĎċĜĤþ ĞĩþĴēĐøĩĚĎċĜĤþĴđđġįţĘČĜĤċ ĂĭĻþĴđţþøĩĚĎċĜĤþĳĒūĐ 2 øĜįţĘČĩĘġİČĚĤĩĢĩĚ ĵċęĶāŤĜİøĒĜĩċįøĤĩęįĳĚīĻĘĎċĜĤþ 1 ĳċĮĤĐ øĜįţĘøĩĚĎċĜĤþĜħ 15 ČĨĞ ĚĞĘĎĨļþĢĘċ 30 ČĨĞ ĎŀĩøĩĚĎċĜĤþĳĒūĐ ĚħęħĳĞĜĩ 40 ĞĨĐ ĘĬøĩĚĴđţþøĜįţĘøĩĚĎċĜĤþċĨþĐĬļ øĜįţĘĎĬĻ 1 ĳĜĬļęþċŤĞęĤĩĢĩĚĕĮļĐćĩĐķĘţĳġĚīĘĵĒĚķđĵĤČīø ĴĜħ øĜįţĘĎĬĻ 2 ĳĜĬļęþċŤĞęĤĩĢĩĚĕĮļĐćĩĐĳġĚīĘĵĒĚķđĵĤČīøĚĢĨġĒĚħČİ 11 ĳĜĬļęþĒĜĩċįøĳĒūĐĳĞĜĩ 20 ĞĨĐ ÿĩøĐĨļĐĶġţĳāĮļĤ ĴđûĎĬĳĚĬęøţĤĵĚûĒĜĩ ûĮĤ Aeromonas hydrophila ĶĐĒĚīĘĩĊĳāĮļĤĜħ10⁸CFU/ml ĜþĶĐđţĤĳĜĬļęþĒĜĩĎĨļþ 2 øĜįţĘ

ĜĨøĠĊħđĚīĳĞĊĶġ ĜĨøĠĊħđĚīĳĞĊĶġ

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ÿĩøĐĨļĐĳĜĬļęþĒĜĩÿĐûĚđ 40 ĞĨĐ ĕđĞţĩĒĜĩĶĐđţĤĂĬĳĘĐČŧĎĬĻĶĢŤĤĩĢĩĚēġĘĳāĮļĤĵĒĚķđĵĤČīøĚĢĨġĒĚħČİ 11 ĳÿĚīĄĳČīđĵČķċŤċĬøĞţĩĒĜĩĎĬĻķĘţķċŤĚĨđĳāĮļĤĵĒĚķđĵĤČīø ĂĭĻþĳĢĺĐķċŤÿĩøĳĒĤĚŧĳĂĺĐČŧøĩĚĳĕīĻĘùĭļĐùĤþĐŀļĩĢĐĨøĳĀĜĬĻę ċĨþ ĴġċþĶĐėĩĕĎĬĻ 2

ėĩĕĎĬĻ 2 ĳĒĤĚŧĳĂĺĐČŧĎĬĻĳĕīĻĘùĭļĐùĤþĐŀļĩĢĐĨøĳĀĜĬĻęùĤþĒĜĩ

ĢĜĨþÿĩøĐĨļĐÿĭþēġĘĳāĮļĤøţĤĵĚûĒĜĩ Aeromonas hydrophila ĶĐĒĚīĘĩĊĳāĮļĤĜħ10⁸CFU/ml ĜþĶĐđţĤ ĳĜĬļęþĒĜĩĎĨļþ 2 øĜįţĘ ĎŀĩøĩĚĳĜĬļęþĒĜĩČţĤķĒċŤĞęĤĩĢĩĚĎĬĻēġĘĴĜħķĘţēġĘĳāĮļĤĵĒĚķđĵĤČīøċĨþĎĬĻøĜţĩĞĘĩĴĜŤĞ ùŤĩþČŤĐĳĒūĐĳĞĜĩĤĬø 20 ĞĨĐ ĵċęĶĐĴČţĜħĞĨĐ ĳĔşĩġĨþĳøČĴĜħđĨĐĎĭøĤĩøĩĚĒŞĞęĴĜħČĩęùĤþĒĜĩĶĐĴČţĜħđţĤ ĕđĞţĩ ĒĜĩĎĬĻĶĢŤĤĩĢĩĚēġĘĳāĮļĤĳġĚīĘĵĒĚķđĵĤČīøĚĢĨġĒĚħČİ 11 ĳČīđĵČķċŤČĩĘĒøČī ĜĨøĠĊħùĤþČĨĞĒĜĩĳĒūĐĒøČīĴĜħķĘţ ĳøīċđĩċĴēĜđĚīĳĞĊēīĞĢĐĨþ ĶĐùĊħĎĬĻĒĜĩĎĬĻĶĢŤĤĩĢĩĚĒøČīĳĚīĻĘĴġċþĤĩøĩĚĒŞĞę ēīĞĢĐĨþùĤþĒĜĩĳøīċđĩċĴēĜ ĎįøČĨĞ ĴĜħČĩęÿĐĳĢĜĮĤĚĤċāĬĞīČĳĕĬęþ 10 ČĨĞ

ġĚįĒēĜ ĤėīĒĚĩęēĜ ĴĜħùŤĤĳġĐĤĴĐħ

øĩĚûĨċĳĜĮĤøĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĬĻĴęøÿĩøēĜīČėĨĊĈŧĤĩĢĩĚĢĘĨø ĂĭĻþĘĬûįĊġĘđĨČīùĤþĵĒĚķđĵĤČīø ĕđĞţĩ ĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĬĻĴęøÿĩøēĜīČėĨĊĈŧĤĩĢĩĚĢĘĨøûĮĤ ĒĜĩĚŤĩ ĴĢĐĘ øįŤþÿţĤĘ ġŤĘĳĎŤĩĞĨĞ ĴĜħ ĢĐţĤķĘŤċĤþ ġĩĘĩĚčĴęøķċŤĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĨļþĢĘċ 52 ķĤĵĂĳĜČ ĂĭĻþĳĜĮĤøÿĩøĵûĵĜĐĬĳċĬĻęĞĹ ĎĬĻ ĘĬđĚīĳĞĊĶġĚĤđĵûĵĜĐĬđĐĤĩĢĩĚ MRS ĎĬĻĳČīĘ CaCO3 ĴĜħĘĬĳĕĬęþ 26 ķĤĵĂĳĜČĳĎţĩĐĨļĐĎĬĻġĩĘĩĚčĎĐČţĤĐļŀĩċĬ ûĞĩĘĳùŤĘùŤĐ 3 ĳĒĤĚŧĳĂĺĐČŧķċŤ ķċŤĴøţ ķĤĵĂĳĜČ ĒĜĩĚŤĩI3 ĒĜĩĚŤĩI4 ĒĜĩĚŤĩI5 ĒĜĩĚŤĩ10 ġŤĘĳĎŤĩĞĨĞI1 ġŤĘĳĎŤĩĞĨĞ I3 øįŤþI1 øįŤþI5 øįŤþI7 øøI1 øøI2 ĞĚ1 ĞĚ2 ĞĚ4 ĞĚ5 ĞĚ6 ĞĚ7 ĞĚ8 ĒĚħČİI2 ĒĚħČİI4 ĒĚħČİI9 ĒĚħČİI11 ĒĚħČİI12 ĒĚħČİI14 ĒĚħČİI15ĴĜħĒĚħČİI16 ĂĭĻþĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĨļþ 26 ķĤĵĂĳĜČĐţĩÿħĘĬĒĚħĵęāĐŧ ČţĤøĩĚĚĨøĠĩġĘċįĜùĤþÿįĜīĐĎĚĬęŧĶĐĚħđđĎĩþĳċīĐĤĩĢĩĚķċŤ ĳĐĮĻĤþÿĩøÿįĜīĐĎĚĬęŧĎĨļþ 26 ķĤĵĂĳĜČ ġĩĘĩĚčĎĐČţĤ ġėĩĞħøċċĨĐĎĬĻĘĬĳøĜĮĤĐļŀĩċĬĶĐĜŀĩķġŤĳĜĺøĎĬĻčİøġţþĘĩÿĩøčįþĐļŀĩċĬĶĐČĨđĶĐĚİĒùĤþ conjugate bile acid ĳøĜĮĤĐļŀĩċĬÿħ

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čİøĢĘįĐĳĞĬęĐ 10 – 12 ûĚĨļþČţĤĞĨĐ ĳĕĮĻĤĎŀĩĒąīøīĚīęĩøĨđĤĩĢĩĚĒĚħĳėĎķùĘĨĐĴĜħĵûĳĜġĳČĤĚĤĜ ĢĜĨþÿĩøĐĨļĐÿħčİø ċİċĂĭĘēţĩĐĳġŤĐĳĜĮĤċċŀĩķĒęĨþČĨđĳĕĮĻĤĴĒĚĚİĒĴĜħĢĘįĐĳĞĬęĐĐŀĩøĜĨđĘĩĶĢĘţĂĭĻþĳĚĬęøĞţĩ deconjugated bile acid ċĨþĐĨļĐøĩĚĳøīċøĚħđĞĐøĩĚċĨþøĜţĩĞÿĭþĘĬđĎđĩĎġŀĩûĨĄĶĐøĩĚĚĨøĠĩġĘċįĜùĤþÿįĜīĐĎĚĬęŧĶĐĚħđđĜŀĩķġŤ ÿĩøĐĨļĐĐŀĩ ĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĨļþ 26 ķĤĵĂĳĜČ ĘĩĎċġĤđûĞĩĘġĩĘĩĚčĶĐøĩĚĎĐČţĤøĚċ pH 2 ĳĒūĐĳĞĜĩ 3 āĨĻĞĵĘþ ĳĕĮĻĤĎċġĤđûĞĩĘġĩĘĩĚčĶĐøĩĚĳÿĚīĄĴĜħĤęİţĚĤċķċŤĶĐĚħđđĎĩþĳċīĐĤĩĢĩĚùĤþġĨČĞŧĳÿŤĩčīĻĐ ÿĩøøĩĚ ĎċĜĤþĕđĞţĩ ĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûķĤĵĂĳĜČĎĬĻĎĐČţĤ pH 2 ĘĬĎĨļþĢĘċ 10 ķĤĵĂĳĜČ ûĮĤ ĚI5 ĒĜĩĚŤĩI5 ġŤĘĳĎŤĩĞĨĞI1 øįŤþI1 øøI2 ĒĚħČİI4 ĒĚħČİI9 ĒĚħČİI11 ĒĚħČİI12 ĴĜħĒĚħČİI16

øĩĚğĭøĠĩûĞĩĘġĩĘĩĚčęĨđęĨļþĳāĮļĤøţĤĵĚû ĳĐĮĻĤþÿĩøĴđûĎĬĳĚĬęĵĒĚķđĵĤČīøĘĬûĞĩĘġĩĘĩĚčĶĐøĩĚ ġţþĳġĚīĘĶĢŤġĨČĞŧĳÿŤĩđŤĩĐĘĬġįùėĩĕĎĬĻċĬ øĩĚęĨđęĨļþĳāĮļĤøţĤĵĚûÿĭþĳĒūĐĒũÿÿĨęĢĐŤĩĎĬĻġŀĩûĨĄĶĐøĩĚûĨċĳĜĮĤøĴđûĎĬĳĚĬę ġĚŤĩþøĚċĴĜûČīû ĎŀĩĵċęĐŀĩĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĎĬĻēţĩĐûįĊġĘđĨČīøĩĚĳÿĚīĄĳČīđĵČķċŤĶĐøĚċ pH 2 ÿŀĩĐĞĐ 10 ķĤĵĂĳĜČ ĘĩğĭøĠĩĒĚħġīĎďīėĩĕøĩĚęĨđęĨļþÿįĜīĐĎĚĬęŧĎĬĻĘĨøĳĒūĐġĩĳĢČįøĩĚĳøīċĵĚûĶĐĒĜĩ 2 āĐīċ ûĮĤ Aeromonas hydrophila ĴĜħ Streptococcus sp. ĵċęĞīďĬ paper - disc diffusion assay čŤĩĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþ øĚċĴĜûČīûġĩĘĩĚčēĜīČġĩĚęĨđęĨļþĳāĮļĤÿįĜīĐĎĚĬęŧøţĤĵĚûķċŤ ÿħĘĬđĚīĳĞĊĶġĳøīċĶĐĚĤđĴēţĐ disc ĎŀĩĵċęĐŀĩĐļŀĩĳĜĬļęþ ĒĚĩğÿĩøĳāĮļĤĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûĘĩĎċġĤđ ĕđĞţĩ ĘĬĳĕĬęþ 7 ķĤĵĂĳĜČĳĎţĩĐĨļĐĎĬĻġĩĘĩĚčęĨđęĨļþĳāĮļĤøţĤ ĵĚû Aeromonas hydrophila ķċŤûĮĤ ĒĜĩĚŤĩI5 ġŤĘČĬĐĞĨĞI1 ĒĚħČİI4 ĒĚħČİI9 ĒĚħČİI11 ĒĚħČİI12 ĴĜħĒĚħČİI16 ĵċę ķĤĵĂĳĜČĎĬĻęĨđęĨļþøĩĚĳÿĚīĄķċŤċĬĎĬĻġįċ ûĮĤ ĒĚħČİI11 ÿĩøĐĨļĐğĭøĠĩĤĐįøĚĘĞīďĩĐĳĕĮĻĤÿŀĩĴĐøġøįĜùĤþĴđûĎĬĳĚĬęġĚŤĩþ øĚċĴĜûČīû ķĤĵĂĳĜČ ĒĚħČİI11 ĕđĞţĩ ĤęİţĶĐġøįĜ Lactobacillus sp. ĴČţęĨþķĘţġĩĘĩĚčĚħđįġĒśāĬġŧķċŤ

øĩĚĕĨĉĐĩûįĊėĩĕĤĩĢĩĚĒĜĩĶĢŤĒĜĩĘĬûįĊėĩĕĳĕīĻĘĘĩøùĭļĐÿĩøÿįĜīĐĎĚĬęŧ Probiotic ĎŀĩĵċęĐŀĩĜİøĒĜĩ ċįøĤĩęįĳĚīĻĘĎċĜĤþ 1 ĳċĮĤĐ øĜįţĘøĩĚĎċĜĤþĜħ 15 ČĨĞ ĚĞĘĎĨļþĢĘċ 30 ČĨĞ ĎŀĩøĩĚĎċĜĤþĳĒūĐĚħęħĳĞĜĩ 40 ĞĨĐ ĂĭĻþĘĬøĩĚĴđţþøĜįţĘøĩĚĎċĜĤþċĨþĐĬļ øĜįţĘĎĬĻ 1 ĶĢŤĤĩĢĩĚĕĮļĐćĩĐķĘţĳġĚīĘĵĒĚķđĵĤČīø ĴĜħøĜįţĘĎĬĻ 2 ĶĢŤĤĩĢĩĚ ĕĮļĐćĩĐĳġĚīĘĵĒĚķđĵĤČīøĚĢĨġĒĚħČİ 11 ĎŀĩøĩĚĳĜĬļęþĒĜĩċįøĳĒūĐĳĞĜĩ 20 ĞĨĐ ÿĩøĐĨļĐĶġţĳāĮļĤĴđûĎĬĳĚĬęøţĤĵĚûĒĜĩ ûĮĤ Aeromonas hydrophila ĶĐĒĚīĘĩĊĳāĮļĤĜħ10⁸CFU/ml ĜþĶĐđţĤĳĜĬļęþĒĜĩĎĨļþ 2 øĜįţĘ ÿĩøĐĨļĐĳĜĬļęþĒĜĩÿĐûĚđ 40 ĞĨĐ ĂĭĻþĕđĞţĩĒĜĩĶĐđţĤĂĬĳĘĐČŧĎĬĻĶĢŤĤĩĢĩĚēġĘĳāĮļĤĵĒĚķđĵĤČīøĚĢĨġĒĚħČİ 11 ĳÿĚīĄĳČīđĵČķċŤċĬøĞţĩĒĜĩĎĬĻķĘţķċŤ ĚĨđĳāĮļĤĵĒĚķđĵĤČīø ĳĢĺĐķċŤÿĩøĳĒĤĚŧĳĂĺĐČŧøĩĚĳĕīĻĘùĭļĐùĤþĐŀļĩĢĐĨøĳĀĜĬĻę ĴĜħĳĘĮĻĤēġĘĳāĮļĤøţĤĵĚûĒĜĩ Aeromonas hydrophila ĜþĶĐđţĤĳĜĬļęþĒĜĩĎĨļþ 2 øĜįţĘ ĕđĞţĩĒĜĩĎĬĻĶĢŤĤĩĢĩĚēġĘĳāĮļĤĳġĚīĘĵĒĚķđĵĤČīøĚĢĨġĒĚħČİ 11 ĳČīđĵČ ķċŤČĩĘĒøČī ĜĨøĠĊħùĤþČĨĞĒĜĩĳĒūĐĒøČīĴĜħķĘţĳøīċĵĚûĴēĜČĩĘĜŀĩČĨĞ (Ulcerative disease) ĶĐùĊħĎĬĻĒĜĩĎĬĻĶĢŤ ĤĩĢĩĚĒøČī ĳĚīĻĘĴġċþĤĩøĩĚĒŞĞę ûĮĤ ĶĐĚħęħĴĚøēīĞĢĐĨþùĤþĒĜĩĘĬġĬĴċþĴĜħÿħĳĚīĻĘĳĒŝŬĤęĳĒūĐĴēĜĴĜħĜĭøÿĐ ĳĢĺĐøĜŤĩĘĳĐĮļĤĴĜħČĩęĶĐĎĬĻġįċĳĢĜĮĤĚĤċāĬĞīČĳĕĬęþ 10 ČĨĞ ēĜøĩĚĞīÿĨęċĨþøĜţĩĞ ĳĐĮĻĤþĘĩÿĩøĵĒĚķđĵĤČīøġĩĘĩĚč ęĨđęĨļþøĩĚĳÿĚīĄùĤþĴđûĎĬĳĚĬęøţĤĵĚûķċŤċŤĞęøĜķøġŀĩûĨĄ 2 ĒĚħøĩĚ ûĮĤ øĩĚĴęţþĕĮļĐĎĬĻĶĐøĩĚęĭċČīċ (competitive adhesion site) øĨđĳęĮĻĤĳĘĮĤøēĐĨþĜŀĩķġŤ (intestinal mucosa) ĴĜħøĩĚĴęţþġĩĚĤĩĢĩĚ ĴđûĎĬĳĚĬęĵĒĚ

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76

ķđĵĤČīøĳÿĚīĄûĚĤđûĜįĘēīĞĳęĮĻĤđįēĐĨþĜŀĩķġŤ ĂĭĻþĒşĤþøĨĐķĘţĶĢŤĴđûĎĬĳĚĬęøţĤĵĚûġĩĘĩĚčĳÿĚīĄĳČīđĵČĴĜħĳĕīĻĘ ÿŀĩĐĞĐķċŤ ĐĤøÿĩøĐĨļĐĴđûĎĬĳĚĬęĵĒĚķđĵĤČīøęĨþġĩĘĩĚčġĚŤĩþġĩĚĒĚħøĤđĎĬĻĘĬěĎďīĿĶĐøĩĚęĨđęĨļþĴđûĎĬĳĚĬę (anti – bacteria compounds) ķċŤĴøţ øĚċĤīĐĎĚĬęŧ (organic acid) ĴđûĳĎĤĚīĵĤāīĐ (bacteriocin) ĴĜħķĥĵċĚĳÿĐĳĒĤĚŧ ĤĤøķĂċŧ (H2O2 ) (Franz et al. 1998) ĳĒūĐČŤĐ

ĳāĮļĤĵĒĚķđĵĤČīøĚĢĨġĒĚħČİ 11 ĎĬĻĴęøķċŤĐĬļķĘţĳĕĬęþĴČţÿħāţĞęĒşĤþøĨĐĒĜĩċįøĎĬĻĶāŤĎċġĤđÿĩøøĩĚČīċĳāĮļĤ øţĤĵĚûĳĎţĩĐĨļĐ ĴČţęĨþāţĞęġţþĳġĚīĘøĩĚĳÿĚīĄùĤþĜİøĒĜĩċĨþÿħĳĢĺĐķċŤÿĩøĐŀļĩĢĐĨøĳĀĜĬĻęùĤþĜİøĒĜĩĎĬĻĳĕīĻĘùĭļĐ ĂĭĻþĳøīċ ĳĐĮĻĤþĘĩÿĩøĳāĮļĤĵĒĚķđĵĤČīøĚĢĨġĒĚħČİ 11 ċĨþøĜţĩĞĘĬûįĊġĘđĨČīøĩĚĎĐČţĤĐļŀĩċĬ ĳĕĚĩħĐļŀĩċĬĘĬûįĊġĘđĨČīĶĐøĩĚ ęţĤęġĜĩęķùĘĨĐĴĜħøĚċķùĘĨĐČţĩþĹ ĶĢŤĘĬĵĘĳĜøįĜĳĜĺøĜþ ġĩĘĩĚčĐŀĩġĩĚČţĩþĹĳùŤĩķĒĶāŤĒĚħĵęāĐŧėĩęĶĐĳĂĜķċŤċĬ ĴĜħĳĕīĻĘùĭļĐ (Jin et al. 1998) ċĨþĐĨļĐĳāĮļĤĵĒĚķđĵĤČīø ĚĢĨġĒĚħČİ 11 ÿĭþĘĬğĨøęėĩĕĶĐøĩĚĐŀĩķĒĕĨĉĐĩĳĕĮĻĤĶāŤĳĒūĐ ĤĩĢĩĚĳġĚīĘùĤþĒĜĩċįøČţĤķĒ

ùŤĤĳġĐĤĴĐħ

ĶĐøĩĚğĭøĠĩûĚĨļþĐĬļĳĒūĐøĩĚğĭøĠĩøĩĚÿĨċÿŀĩĴĐøĳĕĬęþĶĐĚħċĨđùĨļĐġøįĜĳĎţĩĐĨļĐ ĎŀĩĶĢŤķĘţġĩĘĩĚčđĤøķċŤ ĞţĩĳĒūĐġĩęĕĨĐďįŧĤħķĚ ċĨþĐĨļĐĶĐøĩĚğĭøĠĩûĚĨļþČţĤķĒûĞĚğĭøĠĩĶĐĚħċĨđġĩęĕĨĐďįŧ ĳĕĮĻĤÿħķċŤĎĚĩđĞţĩ ĴđûĎĬĳĚĬę ġĚŤĩþøĚċĴĜûČīûċĨþøĜţĩĞ ĳĒūĐĴđûĎĬĳĚĬęġĚŤĩþøĚċĴĜûČīûġĩęĕĨĐďįŧĶĢĘţĢĚĮĤķĘţ

ĳĤøġĩĚĤŤĩþĤīþ

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Eugene, B.C., Sitrin M.D. and Black, D.D. (1996). Gastrointestinal Hepatobiliary and Nutritional Physiology. Pennsylvania :Lippincott-Raven Publishers.

Franz, C.M.A.P., Toit, D.M. and Olasupo, U. Schillinger and Holzapfel, W.H. (1998). Plantaricin D, a bacteriocin produced by Lactobacillus plantarum BEF 905 from ready-to eat salad. Letters in Applied Microbiology. 76: 231-235.

Havenaar, R., and J.H.J. Huis in’t Veid. (1992). Probiotic and Lactic Acid Bacteria in Health and Disease. London : Elsevier.

Jin, L.Z., Ho, M.Y., Abdullah, N. Ali, A.M. and Jalaludin, S. (1998). Acid and bile tolerance of lactobacillus isolated from chicken intestine. Applied Microbiology. 27: 183-185.

Salminen, S., A. Von Wright. (1993). Lactic Acid Bacteria. New York :Marcel Dekkel.

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