HQI NGHrKHOA HQC CONG NGHE SINH HQC T O A N Q U 6 C 2013
PHAN LAP, TUYfiN CHON VA DjNH DANH VI KHUAN DA CO B 6 Dfe PHAN GIAI BA MiA TRONG OlSU KIEN IN VITRO
Vd Van Song Toan, Bo Th[ Cam H u ^ g , Ho Quang Do, Tran Nhan Dung Trubng Bai hgc Cin Tha
T6MTAT
NhSm myen chpn v^ dinb danh vi khuin da co b6 co kh^ nang phan giii ba mla a dieu k i ^ in vitro, 62 dong vi Idiuan duoc phan lap va nhan dien dva vao mgl so dac diem if^"ia" l^c va hinh d^g vi khuin. Tiong d6 44 ddng vi khuin co hoat linh endoglucanase va 30 dong vikfmancd hoat tinh exoglucanas^, Bon dong vi khuan gom d6ngvikhuanBM49 cohoat tinh endoglucanase vi 3 d6ngvi khu9n BM13, BM21, BM97 co ho^ tinh exoglucanase da dugc myen chgn va phoi hgp vdi nhan de khao s^ khi nang phan giai ba mia. Viec ho sung 6% (yh) djch vi khuin BM13, BM21, BM49 voi mat s6 10 t£ baoAnl, li 3 ngay a 38° C frong difiu kifn ky khi in viiro cho thiy ba mia bj phan giaj cao voi ty IS tieu hoa dry matter (DM), neutral detergent fiber (NDF), CF (crube fiber) cao hon so voi nghiem thnc dfii chirng lin hrgt la 27,23,45,09 vh. 42,59% (w/w) KSl qua khulch d^i doan gen 16S rRNA ciia 3 dong vi khuan BMp. BM21, BM49 vdi cap moi 8F vi 1492R bing fcy thu|t PCR va giai trtnh tu bing may giai trinh t\r tv d§ng cho diiy 3 d6ng khuin BM13, BM21, BM49 lin lirgi hiong ding voi cac dong vi khuin JQ923444 Achromobacter xylosoxidans strain BI.6, JQ410786 Bacillus subtilis strain S20, EF530208 Bacillus subtilis strain FS321 6 muc 91%, 94% va 94%.
Tir khda: Achromobacter xylosoxidans, Badllus subtilis,baTfd.z,A^ci)'\x>, invitro MdfiAU
Ba mia ii lo^i phg pham chi>a ham liF^rng cellulose cao vi Id ngu6n thi>c 3n ti^m nang gidu nang lu'O'ng doi vai dgng v|l nhai lai; tuy nhien st^ phan giai chll xo' khong phai do enzyme cOa tuySn tieu h6a mi do ho^t d$ng chO yeu cOa vi khulin sdng trong cac co' quan ligu hda. VI v$y, bS sung vio thi>c Sn cho vat nuoi m$t lirgrng vi khuan c6 khi ning Ihuy phan XO vao h§ Ihong lieu hoa cOa gia siic nhai 1^1 nham giiip gia siic nhai l^i nang cao IJ le phan giai chat xo, lang hpng nhanh, nit ngan thcri gian nuft" dtrang ben c?nh vi|G tan dyng du-g-c nguSn phy phSm ba mia c6 san lam thiic an cho gia siic 6h IhOc diy mgnh noanh chan nuoi bo v6n chu-a phai tri^n b Dong B^ng S6ng Cuu Long noi rigng, ca nuac n6i chung de 6ip img nhu cau dinh duwng cOa con ngu-oi ngay cSng ISng; dong thii viec hu'O'ng toi t§n dyng t a mia lam thO'c Sn cho gia siic nhai l?i cung gop ph^n han chS 6 nhilm hO-u cc cho mfli trufi-ng s6ng do b5 mia gSy ra. Do 66 vi#c Than l^p vi luyen chon vi khuin da co b6 dh phan giai ba mia trong dilu ki|n In vitro ii van 6h can thiel hi?n nay.
' NGUYEN Lieu vA PHl/OWG PHAP Nguygn llfu
Co' ch^t Ba mia dirg'C sliy khfl, nghiln mjn bing m^y nghiln Retsch ZM100 v6i kich thu-oc iubi la 1 mm; Hoa chat;
Carboxylemethyl Cellulose (CMC), H2S04, CaCl2. NaCI, KzHP04, (NI-U)2S04, MgS04. sodium lauryl sulphate neutral (CizHz5Na04S), Ethanol, Pnateinase K,...; Ngu6n vi khuin: djch dg co Ihu o 3 V| tri khac nhau trgn ciing m6t dg co, lr&
vio 6ng falcon yS tning, bao quan tnang loi chuyin v l phong thl nghidm; Thanh phan mai hvcmg nuoi cay vi khuan (Ryckeboer vi dong lac gia, 2003).
Phuvng phap
Phan lip va tuyen chgn vi khuan d|) cb bb
Phuong phap t^ch r6ng vi khuin thyc hien theo c^c buoc: chuan bj mlu, Irai mlu, dy chuyin lilp din khi xu§l hien khuan lgc r6ng, quan s^t d|c dilm khuan Igc, vi khuin v^i 6Anh gia kha nfing Igo vong halo Irgn co chit CMC vS ba mia, Binh gli khi ning phan giii ba mia cua vi khuan
B6 sung 1% (v/v) djch vi khuin mgt so 10^ l^ bSo/ml vio moi trucrng long vcri co chat ba mia. u o 38° C, 3 ngSy trong binh ky khi. M6i 20pl dich vi khuan sau nuoi cly dugc chuyen vao gilng thgch co chit ba mia vbi 6ubng kinh 5 mm d l khao sat v6ng halo; CF cua ba mia sau khi 0 vol vl khuan dypc phan tlch theo phuong phap ciia Weende (1983).
/Chao sit khi ning phoi hgp cua bon dbng vi khuin de phan giai ba mia
B6nd6ng yi khuin BM13, BM21, BM49, BM97 lln lugt ky hieu 1, 2,3 va 4; Thl nghi?m bo tri hoan todn ngSu nhign, 15 nghigm thuc (NT) va 3 lln Igp lai. Vi khuin dugc bo sung v^o cac nghigm IhCc hfong iing nhu sau: NT1(1). NT2 (2), NT3 (3), NT4 (4), NTS (H-2), NT6 (1+3). NT7 (1+41, NTS (2+3), NT9 (2+4), NT10 (3+4), NT11 (1+2+3) NT12 (1+2+4), NT13 (1+3+4), NT14 (2+3+4). NT15 (1+2+3+4); B6 sung 1% (v/v) djch vi khuin mgt so 10' te bao/ml vao moi Inro-ng long, u o 38 C, 3 ngay frong binh ky khi. 20 pl djch vi khuin sau nua cly dugc chuyin vio gllng Ihach co chit ba rfila voi duong kinh 5 mm de khao sil vdng halo lr6n sau khi nhugm voi dung dich Congo Red; lugng xo ba mia con Igi cua moi nghidm Ihiic dugc dOng d l phan tich CF theo phuong phap cua Weende (1983).
Anh hubng cua vi khuan da cb bb phan glii bi mia trong dieu kien in vitro
Thf nghiim bo tri hoan loan nglu nhien, 6 nghiem thO'c (NT), 3 lln Igp lai; trong dd nghiem thOrc ddi chiing (OC) khfing bo sung vi khuan, NT tir 1 den 5 lan iugt b6 sung 2, 4. 6, 8,10% (v/v) djch vi khuin vdi m|l sa 10' l l biolmi vSo mSi nghigm thuc. Thi nghigm dirg-c lien hdnh theo phuong phap in vitro cOa Tilley vh Terry (1963). Cho dung djch dgm vi djch dg c6 vio Ig phun son toi m^u theo ty Ig 4:1. To hgp vi khuin'thich hgp dOng d l chung vao lung nghiem Ihiic theo
' HQI M^HI KH^^ "O"^ ^^f^f^ Mf^np -'^'Mn ^^f^- TriAM nuf^r. 9ni3
mfl tii o h-gn; Sau 3 ngdy l i i n ii b 38° C, Ihu m i u d l danh gia k i t qua thi nghi|m theo cac chl tigu DM (dry matter), CF
—(cnjbe-fibe^NDE:(n6utraLdetergenLfibe[).- - - - - - - - - - Glii trinh ffr vl khuin
DNA ciJa vi k h u i n dugc Irfch ly Iheo p h u o n g phap cOa Maniatis vi dong tac gii (1989), sau 66 6ugc k h u l c h dgi b i n g c^p m i l 8F 5' - AGAGTTTGATCCTGGCTCAiS -3' vi 1492R 5'- GGTTACCTTGTTACGACTT-3' Manialls va dong t^c g& (1989). SSn p h i m PCR d u g c d i ? n di trSn gel agarcise 1 % co bfi sung ethidium bromide (EtBr) vk chyp hinh gel b i n g h$ thing gel Bio-Rad UV 2000 (Hoa KJ); giai Irinh t y b i n g h f fli6ng giai trinh t y t y dong DNA ABI3130, kel qu3 giSI trinh tg 6ugc sir dyng 3h so s i n h Irinh t y trSn ngSn hSng gen (GeneBank) b i n g chuong Irinh BLAST N vi Igp cay p h i h§ b i n g p h l n m i m BioEdil 7.0, SeqVorter, PAUP* v 4 . 0 b 1 0 .
K^TQUAVATHAOLUAN Th^nh p h l n nguyen Hgu ba mfa Bing 1. Thanh phln hoa hpc bS mfa
% DM (w/w) % NDF (w/w) % CF (Ww) % Tro (w/w)
" Chli Ihich: DM: dry matter, fclDF: Nelral detergent fiber, CF: cmbe fiber, w/w; kh6i lugng/khol lugng"
HSrn Iirgng v$t c h i t khd cQa bi mia sau khi s l y b nhigt do 105° C cho d i n khi Irgng lugng khong d l i \i 93,69% (w/w) (BSng 1). Kel quS nSy cho t h i y chi s6 xo Irung linh (NDF) trong ba mia r l i cao so vdi ti 1$ NDF o nhO'ng logi thiic an khdc cOa b6 nhu rom Ida ml IS 774 g/kg, b^t dgu nSnh 158 g/kg, hgt ngfl 267 g/kg (Bueno vi dong tSo gi5, 2005), bfin cgnh do ba mia cho ti If CF 69,83% (w/w) vk tro 1,69% (w/w).
Ket qua phan l$p v & n h ^ n d i ^ n vi k h u a n d^i c o bo
Kit quii da tSch d u p c 62 d i n g \^ k h u i n ; trong dd nhdm khuan Igc trfln co 32 dflng, k h u i n Igc khflng d l u cfl 30 dflng; 6b n i l khuin Igc chia thSnh 5 nh6m: mfl (31dflng), ISi (28 dflng), p h i n g , c l u chong, n i l ho m i l logi chi cfl 1 dflng vi k h u i n ; cfl 3 logl'bla k h u i n Igo. bla nguygn (34 dflng), rang c u a (23 dflng) vi sb Ihuy (5 dflng). MSu sac cCia k h u i n Igc cfl 4 logi:
Irong (7 dflng). trang due (35 dflng), vSng (19 dflng vi cam_(1 dflng). Khi quan sit dac d i l m 62 dong vi k h u i n ta n h j n thiy: 38 ddng khdng cd k l i i nSng di dflng vi 24 dflng vi k h u i n cfl kha nang dl dflng t h l hifn qua kha nang mgc lan r f n g quanh v l l ciy t r f n mfli trufl'ng thgch b i n Iflng. 36 dflng gram gm, 26 dflng vi k h u i n gram duong, Vi k h u i n co 2 dgng:
hlnh c i u (10 dflng) vi hinh qbe (52 dflng); E)a s l c l u k h u i n d l u khflng cfl k h i nang di dflng chl cfl dflng BM7 v i BM91 cfl k h i n i n g dl dflng; kich thu6c vi khi)ln ciia nhflm c l u khuan n i y r i t nhfl chl 10* 0,4 - 0,99 p m . Nhflm vl k h u i n hinh que (10 dflng que d i i v i 42 dflng que ngan) trong do cfl 19 dflng cd k h i nang c h u y i n dflng v i 33 dflng vi k h u i n khdng c d k h i n i n g c h u y i n dgng. 61 ddng vi d u o n g tfnh vfl'i catalase (ky khf khdng bat buflc), chl cd dflng BM35 cfl k i t q u i i m tfnh vfl'i catalase; cfl thl.la dflng vi k h u i n ky khi bat buflc. __^_-
K i t q u i t u y i n chgn vi k h u i n dEi c d b d
K i t q u i cho I h l y 44 trong 62 dflng vi k h u i n co k h i n i n g phan g i i i c o - c h l t CMC (Hinh 1 ) v i 3 3 d ning tgo vdng b^n halo t r i n c o chat ba mfa (Hinh 2),
g vi k h u i n cd kha
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Hlnh 1. Girang kfnh vong halo IrSn cb c h i t CMC;'Cac gid trl.l3 b'ung blnh cua 3 l^n I|p1?l, c i c g i i trj ciing ky t y the hlfn sg khic bl^t khong cd y nghTa th6ng ke b miic 5%
K i t q u i c^o i h l y dflng vi k h u i n BM49 cd kha nang p h i n glai CMC mgnh n h i t vfli dufl'ng kinh Vdiighalo l i 40,3 m m (Hlnh 1), Ichic b l f l cfl j nghTa vdi c i c ' d f l n g vi k h u i n k h i c d m i i c 5%. Ben cgnh dfl, cac ddng vi khuan BM97, BM105, BM103, BM35, BM85, BM119, BM115, BM13, BM59, BM107, BM45, BM21, BM69 cung cho t h i y cfl k h i nang Igo dudng kinh vflng halo dao dflng li> 30,7 mm den 22 m m (Hinh 1).
a 5 IS
M l ^ . ffl^^^iiii tm as
2. B u * n g kinh v6ng halo IrSn ca chat ba mfa; Cdc g l i trUS trung blnh ciia 3 l l n Ifip lai, cSo gid trj c&ng k y l y thlTil^n : khflc bl&t khflng cfl j nghTa thdng k i b muc 5%-
HOI NGH! KHOA HQC COHG NGHE SINH H O C T O A N Q U 6 C 2013
K i t q u i phan tich th6ng ke cho t h i y dflng vi k h u i n BM29 cd k h i nang phan g l i i ba mia mgnh n h l l v o i ducrng kinh vflng halo l i 35mm v i k h i c biet cfl y nghTa Ihong ke b muc 5% vbi d u o n g kinh vflng halo cua c i c dflng vi k h u i n cdn Igi ngogi trir hai dflng vi k h u i n BM13 v a B M 2 1 ; Ben canh dfl, c i c ddng vi khuan BM113, BM83, B M 1 1 1 . BM97, BM57, BM23, BM85. BM45, BM79, cung c h o thay co k h i nang Igo dufl'ng kinh vong halo dao dflng tip 26,3 mm d i n 2 3 mm.
Tflm Igi, qua t u y i n d i g n , 14 ddng vi k h u i n cfl endocellulase mgnh BM49, BM97, BM105, BM103, BM35, BM85. BM119, BM115, BM13, BM59, BM107, BM45, BM21. BM69 v i 12 ddng vt k h u i n cd exocellulase m^nh BM29, BM13, B M 2 1 , BM113, BM83. B M 1 1 1 , BM97, BM57, BM85, BM23, BM45, BM79; trong dfl 5 ddng v i k h u i n : BM13, B M 2 1 , BM97, BM85, BM45 vCra cfl hogt b'nh endocellulase vira cb exocellulase manh.
K i t q u i p h i n giai b a m i a cOa 2 1 d d n g v l k h u i n
K i t qua cho t h i y trong 21 ddng, c d 16 dflng vi k h u i n (hilnh 3) cd k h i nang phSn giai ba mfa; trong dfl dong vi k h u i n BM21, BM13, BM97 Igo d u f m g kfnh vdng halo Ifl'n n h l l ian l u g l l i 31,7 m m ; 31,3 mm va 27,3 m m k h i c bigt cd !^ nghTa t h i n g kg d m i i c 5% vfli dufl'ng kinh vdng halo c i c ddng vi k h i i l n con Igi.
K i t q u i phan tich h i m l u g n g x o cho t h i y sau 3 n g i y nufli d y , cac dflng \ri k h u i n da p h i n g i i i d u g c Kr 1,4% d i n 7,9 % CO c h i t ba mia. K i t q u i p h i n tich (Hlnh 4) cung ch! ra rang 3 dflng vi khuan BM13, B M 2 1 , BM97 co h i f u s u i t p h i n g i i i ba mfa cao n h i t l l n l u g t l i 7,9%, 7,2%, 7,2% v i khac bigt cd •} nghTa b mire 5% vdi 18 ddng vi khuan cdn Igi. Ket q u i n i y phli hgp v d i kel q u i k h i o sat d u d n g kinh p h i n g i i i tren c o c h i t ba mia cua 21 dflng vi k h u i n . Tuy n h i i n , 5 dflng vi k h u i n BM103, BM105, BUtOT. BM115. BM119 khflng tgo d u f l n g kfnh vflng halo tren mfii trudng ba mia nhung h i f u s u i t p h i n g i i i ba mfa cao h o n v i k h i c bigt cfl ^ nghTa b m i i c 5% vfli mau ddi chiing; Tiieo nghien c i i u cua Varel v i d i n g l i e gia (1995), mgl s6 dong vi k h u i n d u g c p h i n l | p ICr da cfl khong p h i t tnen. khuan lgc tdt lr§n m d i tru'fl'ng cellulose nhung co k h i nang lam giam h i m l u g n g cellulose trong mdi trufl'ng nudi c l y . V | y ba (3) ddng vi I d i u i n BM13, BM21, BM97 khflng chi tgo ducrng kinh vdng halo Ifl'n t r i n c o c h i t CMC ben cgnh BM49 (Hinh 1) v i ba mfa (Hlnh 3 ) m i efln cd h i f u s u i t p h i n glai ba m i a cao (Hlnh 4).
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Hinh 3. Qircmg hlnh v&ng halo*
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Hinh 4. Hifu s u ^ p h i n giai ba mfa cua 21 dong vl k h u a n "
• Cdc gid tn la trung binh 3 l l n l$p iai, giS trj cung ky ty the hl#n sy khdc bigt khflng cfl y nghTa lh6ng kS b mire 5%; %CV = 4,59%
' • Cdc gid trj Imng blnh cua 3 tin lap l^i. gia trj ciing ky ty thS hien s y khac blfl khflng c6 y nghTa thflng k§ b mdc 5%; %CV = 7,08%
Ket q u a phoi hg>p b d n d d n g v i k h u i n
K l l q u i d i o t h l y NT1 (BM13), NT2 (BM21). NT3 (BM49) va NT4 (BM97) tao d u d n g kinh vflng halo nhd so v d i mOt s6 nghifm ttiuc cd p h l i hgp c i c dflng vl k h u i n v o l nhau n h u NT5 (BM13 v i BM21), NT6 ( B M 1 3 . v i BM49), NT7 (BM13 v i 6M97) cfl dufl'ng kinh vflng halo I i n l u g t l i 23,7 mm; 23 mm; 21,7 mm (Hinh 5) va cao hon d u d n g kfnh vdng halo ciia NT1. NT2, MT3; tuy nhign NTS (BM21 v i BM49), NTS (BM21 v i BM97) va NT10 (BM49 v i BM97) Igl cho k l l q u i d i r d n g kinh vdng halo nhfl hon NT1 v i NT4; Td h g p BM13, BM21-va BM4 (NT11) cfl dudng kinh vdng halo l d n n h i t 29 m m v i khac bift cfl^y nghTa gi&a c i c nghifm thiic 6' m i i c 5%; Cho I h l y cd I h l cd s u tuong t i c Ifch eye hogc i i c c h l g i i i a c i c dflng vl khuan v d i nhau. BM13 v i BM21 c d kha n i n g sinh exoglucanase mgnh p h l i hop v d i BM49 c d kha n i n g sinh endoglucanase mgnh nfln p h l i h g p 3 ddng vi k h u i n BM13, BM21 v i BM49 sinh enzyme p h i n g l i i ba mia mgnh n h l l ; Kel q u i (Hlnh 6) cho I h l y NT11 l u g n g DM m i t dl n h i l u nhai l i 15,95%, m f c dii k h i c biet khdng y nghta vdi NT4, N T 5 , NT6, NTT nhung k h i c b i f t c6 y nghTa v d i cac nghigm Ihiic efln Igi fl' mire 5%. Ben canh do. t o h o p 3 dflng vi k h u i n BM13, BM21 v i BM49 (NT11) cung cfl k h i nang tgo d y d n g kinh vflng halo Idn nhat (Hinh 5). Theo f r i n C i i (1979) hogt d f n g k l l hgp cua endoglucanase v i exoglucanase se d i n d i n thay ddi dac d i l m be m i t cua p h i n tCr cellulose t i i d o se l i m cho ly I f p h i n giai thay d l i nhanh chflng.
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Hlnh 5. D i r m g kfnh v6ng halo*
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Hlnh 6. OM gidm**
• Cdc gid trj tmng blnh 3 l l n ldp Igi. gid tij ciing ky l y t h l hi^n su- khac blfl khflng c6 y nghTa thing ke d miic 5%; %CV = 6.22%
* Cac gid tri tmng bmh 3 lan l$p lai. gid tri ciJng ky ty the hign sy Wiac bi#t Wiong cd y nghTa B ^ ke d miic 5%; %CV =
tHQl-MGHI KHOA-HQC OOW
P h i n g l i i ba m l a ciia v i W i u i n b dieu k i f n in vitro ^ ^
Kit qu"3"(Hinh 7) fhrf I h l y N T ' ^ ••'^"^ f ' ^ - " ^ " t"^« ^^ NHF, CE-Cao nhai lan lim-l la 27,93, -45,09 va-49,fiqy„ (w/w) i^hac b l f l CO J nghTa thdng ke b miic 5% so vfl'i nghifm thiic DC, N T 1 , NT2, Gall v i Huhlanen (1951) cho b i l t d l i vdi qua trinh tiflu hfla nhai Igi, dieu quan hgng khflng chi la ty I f cac l o i i vl k h u i n frong dg cd ma l i so lugng cOa chiing hay long s i vi k h u i n . Vi the, cd I h l d KTZ dgt d u g c tdt n h i t ve t J le cSc loai vi k h u i n frong dg cfl v i c i t i n g so vi W i u i n nen cho k i t q u i rt if t i i u hfla cao n h i i NT4 va NTS mgc dO d u g c b l sung lugng vi k h u i n Idn hon nhung cfl t h l da l i m i n h - h u d r g den c i n bang hg vi ^ n h v g l Irong^dg c d nSn khSnang figu hfla kem h o n NT3. Theo Bryant v i Burkey (1953) cho bilt hg vi sinh v f l dg cfl la mflt h f t h i n g d u g c c i n bang i l l , s y b i l n d l i l i n g so ciia h f n i y it phy fhugc v i o flgc Ifnh thiic i n nhung quan h f n h i l u den m i i c d f pha l o i n g c h i t chiia djch dg co sau khi an, d$c d i l m nufli d u o n g v i tdc dg sinh san giOa nhfing nhdm vi sinh v i t .
J<lt q u i cho I h l y vdi v l f c b l sung 6% djch vl k h u i n t l hgp 3 dflng vi k h u i n BM13, BM21 v i BM49 v d i mgt so 10^ le bio/ml thl kha n i n g Hdu hfla thiic an d bfl l i t n h l l vl v f y c d t h l i m g dyng t i hgp 3 dflng vi k h u i n n i y b l sung vao k h i u phln thiic i n x o cOa bd nham giup bd cai thign I d i i nang tigu hda c h i f x o d l gdp phan cho bfl tang lipng nhanh.
Nghifm thiic
Hlnh 7. Ty If lieu hda DM, NDF, CF; Cdc gia trj trung blnh ciJa 3 lan l^p lai, gia ti^ cfing ky t y thi h i f n s y khdc biet khdng cfl y nghia Ihong ke b mire 5%; %CV(DM) = 5,17%; %CV(NDF) = 3,61%; % CV(CF) = 4,87%
K i t q u i d i f n d i san p h i m PCR
K i t q u i p h i n i i n g PCR vdi c f p mSi 8f v i 1492r ciia 3 dflng vi k h u i n BM13, BM21, BM49 (Hinh 8)
Hlnh 8.BI$n di sap phim PCR; Gi^ng 1: thang Ikb; Gilng 2: d i i chiing dm; Gllng 3: BM13! Gieng 4: BM21; Gieng 5: BM4g K i t q u i g l i i t r i n h t u
K l l q u i g i i i trinh t y cho I h l y ddng vl khuan BM13 tuong dong vfln JQ923444 Achromobacter x^osoxldans strain BL6 ' (China) b mine 9 1 % . Achromobacter xylosoxidans cfl k h i nang p h i n g i i i hemicelluloses v i cellulose, sau 14 n g i y nufii trong mdi trudng nufli c l y l i m giam 67,3% cellulose va 73,5% hemicelluloses; Dflng vi khuan BM21 v i BM49 l l n l u v t lugng ddng vdi ddng JQ41786 Bacillus subtilis strain S 2 0 (Cameroon) v i EF530208 Bacillus subtilis strain FS321 (China) d mire 94%. Bacillus subSlis l i vi k h u i n hinh que, gram dirong, dugc phan I f p l i i dg cd I r i u bfl (Sivers v i d i n g . - l i c g i i , t 9 7 7 ) ; t ! i n j g l c i ( R a y v i d l n g t a c g i i , 2 0 0 7 . ) ; - " - - -- — — - — • - • - - - . - -•--— ^ - -
K i t qua p h i n t i c h pha h f
• Trinh t y cCia 3 dflng vi k h u i n BM13, B M 2 1 , BM49 v i -6 dflng vl k h u i n Bacteroldes mmlnicota, Rumlnococcus albus, Fibrobacter succinogenes, Rumlnococcus fiavefaclens, BiAyrtvibrio ffbrisolvens v i Lachnospira maltipara cd k h i nang phSn^giii cellulose cao 0<amra, 2005) d u g c diing d l p h i n tlch p h i hg (Hinh 9). Dflng vi k h u i n BM21 v i BM49 cfl quan h f r i t g i n (bootstrap l i 100%), BM21 v i BM49, Butyrivibrio fibrisolvens vi Lachnosdra multipara cOng cfl quan h f r i t g i n vdi nhau (bootsh-ap l i 99%); T h i m v i o dd, 4 dflng vi khuin n i y cd quan h f g i n vdi ddng vi k h u i n BM13 vfl'i chl s6 bootstrap l i 100%. N g o i i ra fium/ndcoccus flavefadens, Fibmbacter .sucdnogenes, Rumlnococcus albus vi Bactemides rumlnicota) cfl quan he xa hon vfli
Hinh 9. Gian do pha he; Cdc cTiI sd Cl = 0.7361; Rl = 0,5666; chi s4 bootstrap ffu^c ^ i trSn d^u cdc nhdnh Irong gidn dfl
BM21, BM49. Butyrivibrio Sbrisolvens. Lachnospira muWpara va BM13.
H O l N G H I K H Q ^ J l g C ' C O N G N G H ^ I j i l H HCit j | ^ ^ ( ^ ^ 2 0 1 3
K 6 T LUAN
Trong tong so 6 2 d6ng vi k h u i n d? O 0 i § O v v c phan l | p , co 44 d6BS vi Idiu^ ^ . -*, , - „ r .,a ^ ^ d6nQ vn khuSn CO l i h i nang phan giai bSmra; ' ^ i , tenangphangiSICMCvasJoongvn T 6 h p p 6% (v/v) cua djch 03 dong vi'i^n ( B M t 3 , BM2 va BM49) J a , w o c . uA A ^ ^ 4 „ 5 n n ohSn QiSi mgnh ba mia trong dieu k i j n in vilm<m l m cho I J IS tlsu hoa DBI, t t f i ( - ! ' G ^ • * ? " * ° 1 ' S , i i nShVam Ihiic doi chi/ng khSc biet cd > nghta th6na J * » .rfftc 5 % lan lirot la 2 7 , 2 3 . 4 f c & ! « » n l ^ ' " ^ cao hon v H «£n ngniem KSt qua giai trinh tLP 3 dong vi khu,S^fiM13, B M 2 1 , BM49 ian l i m ^U^Offi^ . . .^ . - w , „ S n tnq23444 /ichromotecterxyfosoxWans strain S « l ; J 6 4 1 0 7 8 6 B a c f c s ^ " ' ^ H ^ i ^ ' l l ^ l S ^ ^ ^ S S M ^ ^ F S 3 Z 1 v c S i m i i c a o n g h l n h l a 9 1 , 9 4 » 4 # « « . ' - a S v 4 EF530208 S a c l l M suonis s m i n
T A ) u e u T H A M K H A o
S I . J w J s L f a S " ' " " ' ' ' " " * * ^ " " ' ' * = " " ' " " « ' ^ " " V f : ° ' » * - 4 nummus group, of b . d o r l , in tho bolno
m . S ; S ^ f c i ; s l T o ' s " " ^ ' * " " ' " " ' * * " ' ' ••• ° ° " " ' ' " " " " " • " ' ^ f i ^ ' , - a o e ot inoculum »«,rc. in . aa. producHon Gall LS. Huhtanon CN (1951) Rumen Orggrfems. J. Dairy Sd. 34:353-392.
Kamra DN (2005) Rumen microbial ecosystem. Curr Sd 89(1).
ManJallsT s.mbook J. Erilsek EF (1989) M * a , i „ | a b o r , l o n - manual. '^.S/I.SihSpiivluMorLabeMay. CM Spring H>*or. NV
S S i * S i ; J = S l 4 t , 2 ° X - '• ' * ' • ' '™' *'^»«*«' » ^ »< «<»* '""»= •»"'•'* '" •
n i l e , JMA. Tony RA (1963) A i ™ stag, l o c h . ^ i s fcr In v«» dig.slon <'Ws^t,cf,.J. © , . Grassland Soc 18:104-111.
_T,4n CO (1979) Smh i j v i hCa sinb IBu hoa c h d , „ o . J l nhai 1,1. mi »"*SS» itessfc^^ ^ K ; I W I W W/. Ir.35 - 1 2 0 . S b S l " l " - l ' ? r J ' " ' ~ " " " " " * " " " " ' ' * " ' " ° " * ° * * « ? . « • > . ft*, „ m . n and pig Infainei tea. Appl E n — Weende (1983) Fwe extraction unit for detorininidg raw fiber content Velp §ggo(j.w , c j ^ „
sivereVS.TopdiiT MP, Kovalevskala TM (19771 Fomiatlon of organic acWafcy^,. , . „ .=rt„.,c onrf n=ri«us subflBS Isolated fmm the mm.n of cattle. MlmUol. a , »S(1 ):31.33. " « " »» M r * of SacJIos mesenteHcos and B.ollus s r a ~ Ray AK. Bairagi A, Saritar GK. Sen SK (2007) Qpllmlzallon of femionbatlon efibdWfiB. * « . . j i . „„i„.i,r,n h„ narWi,^ suWHis CY 5 and e.cJ/us o/na./ons TP3 Isolated 1mm fish g l l , Mla Idtlh/d PScaf 37(1); 3 ^ * " ' " ' " H " ! " " producUon by BeclBus suoms >-
ISOLATION, SELECTION AND "^IDENTIFICATION OF BACTERIA ISOLATED FROM GASTRIC JUICE OF A COWS RUMEN FOR HYDROLIZATION OF SUGARCANE BAGASSE WWTRO niui^v^i.
V o V a n S o n g T o a n ' , Do T h i Cam H u o n g , l i d O u a n g Do, Tran Nhnii Uy=
Can Tho Unh/Br^ity'
For selecting and identifying mmbant baoterltl bvvn capacity for hydroiysu „r „„„^ ,. . „«,„, „nnrHtlort Sixty twul strains of bacteria were isol.ted fiom gaslrl«,Jl!te of. cow's rumei u d l k - i f ' " • " " " ' " ^ ' T ^ / J J ™ 1 m o i j chsrsctedsti^, b.cted. and colony .pp..,.„4, Mte, .creeping tl«se b » S ' Ti^^ ^ 0 ^ ^ t o s, 1 S S S 3 ..d08lnc«me «aivity with drfttnint di«a,l,|, „t Wlo aones on CMC l S r i „ X ! ™ 1 w , ^ t e o of s^ty " o > " * ' bsderi. bad exoEluc«»se ectivity wilh vartws aUmeters of halo zones o S ? ' . ' " ? " ' ° " ' * " ? , ™ ° Fom rf iMm i « ^ smdlts including one strain (BM49) with stwoj woglucanase icirity ffl 4 f c . i t ' S l B « ' end B M s T " * ^ exoglucanase activity were tested theff symbiiMSS in a hvdrolvtic process of'^\.-\; . " ^ ^ (BMI3, BWUl ono «'^^'J , ^ r « s » s ^ „ s i o n o f d ™ , L i n s B 1 ^ 1 3 , B M 2 L i l " C « , r t o S v . a . S ^
38- C were able to hydrolyxe sugarcane baggSB, isbstate with a high d i g S S f e S J fe'SS H ?^ 1 ^ 7 M l ( « M » H (w/w) and 42.59 % (w/w). The result cf am^iftsas »= 16S rTOA gSe otTw i ' ^ r S L ' j L ^ S s B ^ * 5 ^ pair of primeni 8F and 1492Rby PCR and sis(deli„g by autoiaued s = < I » S ? ^ S , v v n ' S * . l , w sJato ffll BM13, BM21. BM49 wero similar to J t J s S S l A ^ Z i o t ^ ^ ^ 7 ^ ^ ^ * ^ ^ ^ ^ ^ J ^ , M ^ M EF530208tel//..»M/ttstn,nFS321 will, ,te was ido.tityof91%, 9 4 ^ l ^ ^ ^ ' j i y ' ' * ' • ' ' " " • " ' " " ' ^ M Keyword, • Achronwbacier xylosmidans, Bav^,.s itiAlilis, baggase, cows' fHHHgri. invllfo
•Aulborfor conespondence: Tel: « 4 710 3 KSSSMiWoMIe: '84 9B89SB4aaitey,s«; vvJKilnftau edu.vn