Nghin cu sn xut khng sinh vancomycin t x khun Streptomyces orientalis Th hai, 06/06/2011, 15:49 GMT+7

Vancomycin l cht khng sinh c s dng rng ri v c tc dng tch cc trong cha bnh. Cc nh khoa hc Vin Cng ngh sinh hc (Vin Khoa hc v Cng ngh Vit Nam) ang nghin cu iu kin thch hp sn xut vancomycin t bin chng x khun Streptomyces orientalis nhn c t x l N-methyl-N-nitroN-nitrosoguanidin (MNNG) ln t bo trn ca chng gc. Vancomycin l cht khng sinh thuc nhm glycopeptid c tc dng tch cc trong iu tr bnh, tng c coi l phng thuc cui cng v c kh nng iu tr c cc bnh nhim trng nguy him do cc chng vi sinh vt khng methicillin (cht khng sinh nhm -lactam) gy nn. Vancomycin c a vo cha bnh t hn 40 nm qua, nhng ngy nay vn c coi l khng sinh quan trng do hiu qu cha bnh cao khi dng mt mnh hoc phi hp vi cc khng sinh khc, chng li cc vi khun nhn vi nhiu loi khng sinh thng dng. Bi vy, vic nghin cu sinh tng hp vancomycin vn c quan tm, pht trin, t hnh thnh nn th h khng sinh mi c hiu qu cha bnh cao. Hn na, nghin cu ln men vancomycin v nm vng quy trnh sn xut cht khng sinh ny cn to tin cho vic xy dng c s sn xut cc cht khng sinh quy m cng nghip trong iu kin Vit Nam, gp phn thc hin mc tiu ti nm 2020 sn xut c 50% tng s thuc, do B Y t ra. Trong bi cnh nh vy, vic nghin cu xy dng quy trnh sn xut vancomycin bng nguyn liu trong nc, ph hp vi iu kin kinh t v mi trng kh hu ca Vit Nam l cn thit. Cng vi n, vic trin khai xy dng mt c s sn xut khng sinh ny vi cng sut 500 kg/nm gp phn phc v cng tc chm sc sc khe cng ng. Chng ging vi sinh vt Chng x khun Streptomycws orientalis 4912 v cc chng vi sinh vt kim nh Bacillus subtilis ATCC 6633, B. cereus ATCC 21778, Staphylococcus aureus 209P, Sarcina lutea v Eschrochia coli PA2 (B su tp ging ca Phng Cng ngh ln men, Vin Cng ngh sinh hc); cc ha cht dung phn tch, nh lng v vancomycin chun (Merck) v cc mi trng nghin cu l Gause 1, A4, A-4H, TH447, A12, A-9, 48. La chn mi trng iu kin ln men sinh khng sinh ca chng S. orientalis 4912 Th nghim ln men trn mt s mi trng thng dng trong ln men sinh khng sinh x khun cho thy, chng S. orientalis 4912 c hot tnh khng khun mnh v la chn c mi trng MT48 cho hot tnh khng sinh cao nht c th lm mi trng c s nghin cu nh hng ca cc ngun dinh dng v iu kin ln men n kh nng to khng sinh. Chng S. orientalis 4912 s dng tt ngun ng saccharose vi hm lng thch hp l 3%, cho hot tnh khng sinh cao. Trong s cc ngun nit th nghim th bt u tng cho hot tnh khng sinh cao nht, vi hm lng 0,2% l thch hp. Nghin cu nh hng ca nhit v pH cho thy, nhit thch hp cho sinh trng, pht trin v sinh tng hp khng sinh ca chng S. orientalis 4912 l 280C v pH t 6 n 8. Lng ging c cy vo mi trng ln men thch hp 6 - 8 % so vi mi trng ln men. Nghin cu ng thi qu trnh ln men chng S. orientalis 9412 cho thy, sinh khi v hot tnh khng sinh tng dn v t cc i sau 120 gi ln men. Nh vy, ng

thi qu trnh ln men chng ny c c trng ging nh cc chng x khun sinh khng sinh khc. Nng cao hot tnh khng sinh ca chng S. orientalis 4912 Trc ht, cc nh khoa hc Vin Cng ngh sinh hc p dng phng php gy chng t bin bng tia UV. Kt qu nghin cu kh nng sng st ca t bo trn v bo t chng S. orientalis 4912 sau khi x l UV sng st t 1-10%, kim tra hot tnh khng sinh theo phng php cc thch cho thy, kh nng sinh tng hp khng sinh ca chng ny tng ln t 8-30,3% i vi x l bo t v 66,33% i vi x l t bo trn. Sau , cc nh khoa hc tip tc p dng phng php th hai, l gy chng t bin bng MNNG. Trn c s la chn nng MNNG, pH v thi gian x l thch hp x l bo t v t bo trn th t l bin chng c hot tnh khng sinh cao hn chng gc l 80,8 v 92,86 %. Kt qu nhn c bin chng S. orientalis 4912-81-61 (x l t bo trn bng MNNG) c hot tnh khng sinh cao nht l 1683 mcg/ml, c la chn cho nghin cu iu kin ln men sn xut vancomycin (hot tnh khng sinh chng gc l 866 mcg/ml). La chn mi trng v iu kin ln men vancomycin ca chng t bin Da trn mi trng ln men thch hp cho chng S. orientalis 4912, tin hnh la chn mi trng ln men ti u theo phng php quy hoch thc nghim ca BoxWilson cho bin chng S. orientalis 4912-81-61 nh sau Saccharose 49,8 g/l; glucose 17 g/l; bt u tng 30,6 g/l; NaCl 2,5 g/l; CaCO3 2 g/l; CaCl2 40 mg/l; CuSO4 10 mg/l. nh hng ca cc yu t iu kin ln men cho thy, t l tip ging 4%, nhit ln men ti u 28oC v nng pH thch hp 7. Nghin cu nh hng ca hm lng oxy ha tan (dO2) trong mi trng ti s sinh trng v sinh tng hp vancomycin, cc th nghim ln men bin chng S. orientalis 4912-81-61 c thc hin trong h thng Bioflo 110 dung tch 7,5 lit vi cc iu kin pH, nhit v t l ging xc nh trn. Kt qu xc nh sinh khi kh v hot tnh khng sinh sau 120 gi ln men cho thy, khi dO2 c duy tr mc 20-30% th lng vancomycin v sinh khi t cao nht, tng ng bng 2983 mcg/ml v 9,8 mg/ml. Nh vy, kt qu ca cc th nghim ny cho thy vic cung cp oxy ha tan v bo m o trn tt trong bnh ln men l iu kin thit yu ca sn xut vancomycin. xc nh thi im thu hi vancomycin thch hp nht, bin chng S. orientalis 4912-81-61 c nui trong thit b ln men Bioflo 110 dung tch vi thnh phn mi trng ln men ti u cho thy, bin chng ny pht trin tt trong bnh ln men, c bit t gi th 48 sinh khi ca chng tng nhanh t 5,8 mg/ml. Cng ti thi im ny chng bt u sinh vancomycin, ti 120 gi nng t cc i l 2983 mcg/ml. Tch chit v tinh ch vancomycin t dch ln men Kt qu kim tra vancomycin tch chit t bin chng S. orientalis 4912-81-61 bng sc k lp mng trn h dung mi Butanol - axit acetic - H2O (4 : 3 : 7) cho gi tr Rf ca cc mu l 0,75; bng phng php ph khi Agilent 6310 Ion Trap trn my HPLC-MS, trng lng phn t l 1449,27 v bng sc k lng cao p trn my HPLCSPA-10 Shimadzu, thi gian lu l 4,4 pht ging nh vancomycin chun (Merck). Sc k HPLC cho thy khng c cc pic tp chng t vancomycin ch phm kh tinh sch, tinh khit t 95,4%. Vancomycin l mt khng sinh nhm glycopeptid c s dng cha cc bnh nhim trng do cc vi khun Gram dng gy ra, c bit l cc vi khun khng li khng sinh methicillin v penicillin. Kt qu nghin cu xy dng c quy trnh thch hp sn xut vancomycin t bin chng nhn c cao hn chng gc 344%. Cht khng sinh thu nhn c t dch ln men tng ng vi vancomycin chun (Merck
. M U Vi sinh vt gy bnh nh hng rt ln n nng sut v tnh n nh trong nui thy sn. kim sot vi sinh gy hi, phng php truyn thng l dng cc ha cht dit khun v khng sinh, tuy nhin cc ha cht dit khun

nh Chlorin ngoi vic lm tng mt Vibrio sau khi s dng, cc dn xut ca chlorin cn l nhng cht gy t bin v ung th nh : Chloroform (CHCl3), bromodichloro-methane (CHBrCl2), dibromodichlo-romethane (CHBrCl2), v bromoform (CHBr3). Cc loi khng sinh to ra cc chng khng khng sinh, cc plasmid m ha cho cho cc gen khng khng sinh s truyn t vi sinh vt gy bnh thy sn sang cc vi sinh vt gy bnh cho ng vt v ngi. V l ngy nay nhiu loi khng sinh b cm s dng trong nui thy sn nc ta II.KIM SOT SINH HC TRONG NUI THY SN Thut ng probiotic c Parker a ra nm 1974. Tc gi nh ngha probiotic l nhng sinh vt hoc nhng cht gp phn lm cn bng h vi sinh vt ng rut. Thc ra tng s dng vi khun probiotic c Elie Metnhicoff a ra nm 1907. Khi kim tra vic tiu th sa chua tc gi tm thy nh hng vi khun Lactobacillus delbrueckii ssp bulgaricus n vic ko di tui th ca ngi Bungary . Ngy nay probiotic c s dng kh hiu qu phng bnh cho ngi v gia sc, gia cm trn cn. Nhiu vi sinh du?c s dng sn xut ra cc ch phm ng dng rt hiu qu nhm kim sot cn trng gy hi cho cy trng nh vi khun Bacillus thuringiensis, nm Beauveria bassiana, Metarrhizium anisopliae, virus NPV.v.v. Ngoi ra cc ch phm vi sinh cn c s dng lm phn bn vi sinh nhm phn gii cc cht hu c lm giu cho t, phn gii ln kh tiu thnh ln d tiu cy trng hp thu c. Cc ch phm vi sinh s dng trong cc h thng x l rc thi v nc thi, lm sch du m trong cc v trn du. Mc d vy, vic s dng cc ch phm vi sinh trong nui thy sn vn cn kh mi m. Cc ch phm s dng trong nui thy sn hin nay c th chia lm 3 loi: -Cc ch phm c tnh cht probiotic gm nhng vi sinh vt sng nh cc vi khun thuc ging Bacillus, Lactobacillus, Saccharomyces ngi ta thng trn vo thc n hoc qua trung gian nh Artemia, Rotifer. -Loi th hai gm cc vi sinh vt c tnh i khng hoc cnh tranh thc n vi vi sinh vt gy bnh nh vi khun Bacillus licheniformis, Bacillus sp. Vibrio alginolyticus -Nhm th 3 gm cc vi sinh vt ci thin cht lng mi trng nh vi khun Nitrosomonas, Nitrobacter, Actinomyces, cc loi Bacillus khc nhau, cc loi to, cc vi khun ta khng lu hunh nh Rhodobacter sp. Rhodospirillum, Rhodopseudomonas viridis, Rhodopseudomonas palutris, Rhodomicrobium vanniell, cc loi nm Aspergillus oryzae, Aspergillus niger, Rhizopus sp Tuy nhin c nhiu chng vi sinh vt thc hin c nhiu chc nng khc nhau nn ranh gii gia 3 nhm ny i khi khng c phn chia r rng, v vy ngy nay nhiu ngi gi chung cc ch phm vi sinh s dng trong nui thy sn l probiotic Trn th gii c kh nhiu cc nghin cu v vic s dng probiotic trong nui thy sn : Maeda v Nagami (1989) trnh by phng php kim sot sinh hc trong nui thy sn , Maeda v Liao (1992) trnh by hiu qu ca ging vi khun tch t bn trong b nui u trng tm s P. monodon. T l lt xc v sng st ca u trng cao hn trong cc l th nghim so vi i chng Cc th nghim s dng probiotics u tin trong nui tm do cc nh nui tm ging thc hin nhm tm cch ci thin mc vibrio c li trong cc b nui gi l sucrose fermentors . thc hin mc tiu ny ngi ta s dng ng n cho vo trong cc b nui tm ging kch thch s pht trin ca Vibrio spp. c li (cc chng ln men ng saccha-rose). Tip theo cc chng vi khun Vibrio c li c nui cy tng t nh phng php nui to cho vo cc b nui tm ging. Vi to Tetraselmis suecica c th km hm vi khun gy bnh c nh Aeromonos hydrophila, Aeromonas salonicida, Serratia liquefaciens, Vibrio anguillarum. Chng Vibrio alginolyticus khng gy ra bt k hu qu c hi no cho c hi. S dng phng php iu tr cho, probiont c tha nhn l km hm tc nhn gy bnh trn c nh V. ordalii, V. anguillarum, Aeromonassalmonicida v Yersnia ruckeri. Chu c nhiu nghin cu s dng cc ch phm vi sinh trong nui tm, c

bit Thi Lan. Jiravanichpaisal & ctv (1997) s dng Lactobacillus sp. trong nui tm s (P. monodon Fabrius) . Trung Quc, nghin cu probiotic trong nui thy sn c tp trung vo vi khun quang hp. Qiao Zhenguo & ctv (1992) nghin cu 3 chng vi khun quang hp s dng cho tm (P. chinensis) bng cch cho vo thc n hoc cho v nc nui tm cho thy c s gia tng kh nng pht trin ca tm, loi tr nhanh chng NH3-N, H2S, acid hu c v nhng cht c hi, ci thin cht lng nc, cn bng pH. Zhermant & ctv (1997) cho bit trong rut ca u trng protozoea c nhiu vi khun ca mi trng xung quanh nn c th gy tr ngi cho cc th nghim probiotic. Khi nui chng vi khun probiotic trong b vi u nauplli tm Litopenaeus vannamei vi mt 103 t bo/ ml th ngn cn c s xm nhim cc vi khun gy bnh ngay nng 107 t bo/ml . Nm men v nm mc cng c s dng ci thin t l sng v nng sut u trng tm Litopenaeus vannamei (Intriago et al., 1998). Nm men c sc t (Rhodotorula) v nm mc phn hy chitin c tch t mi trng bin s dng, tuy nhin c rt t thng bo v kt qu ng dng ]. Vic phn lp cc chng vi khun probiotic l vic lm mang nhiu tnh cht kinh nghim, t c s khoa hc, v vy c v s cc nghin cu v probiotic b tht bi, iu c th l do vic la chn cc chng vi sinh khng thch hp. Cc bc la chn c xc nh, nhng cn phi thch ng vi tng k ch v tng mi trng. Cn phi hiu c ch hot ng ca probiotic vch r tiu chun chn lc cc probiotic hu hiu . Vit Nam nhng nghin cu v vic s dng cc ch phm vi sinh ci thin mi trng nui tm hoc c tc dng probiotics con tng i t . Trong nhng nm gn y B Thy sn cho php lu hnh sdng nhiu ch phm vi sinh , nhiu ni lm quen vi vi vic s dng cc ch phm vi sinh ny v c kt qu kh tt, tuy nhin cn c mt s nh gi ton din v hiu qu kinh t v phng php s dng. Bng 2 : Tm tt cc th nghim invitro kim sot ca V. harveyi Kim sot sinh hc Tc nhn p dng Liu s dng Hiu qu Ti liu Bacillus S11 Probiotic/ Cho vo thc n T l sng 74% Phianphak & ctv, 1997 L. casei L. acidophilus Probiotic/ Cho vo thc n c ch pht trin Jiravanichpaisal & ctv, 1997 V. anginolyticus Nui ng thi V. anginolyticus/ V. harveyi 1:10 cfu c ch pht trin 0 to 100% Ruangpan & ctv, (in press) Chlorella sp. Nui ng thi 1000 : 1 t bo / cfu c ch pht trin Direkbusarakom & ctv, 1997 Skeletoma costatum Nui ng thi 500 : 1 t bo/cfu c ch pht trin 35 to 100% Panichsuke & ctv, 1997 Dch chit l i Cho vo thc n Nng c ch ti thiu 1250 mg/ml c ch pht trin Direkbusarakom & ctv 1997 III. KT LUN 1. S dng cc ch phm vi sinh trong nui thy sn ni chung v trong nui tm ni ring nc ta l mt xu hng tch cc v ngy cng m rng. C s khoa hc ca vic s dng cc ch phm vi sinh l to c s cn bng gia sc khe ca ng vt nui tt, mi trng c ci thin v s lng vi sinh gy bnh c khng ch. 2. tng cng hiu qu s dng vi sinh trong nui thy sn cn y mnh cc nghin cu v mi quan h gia cc vi sinh vt probiotic v tng i tng nui, gia vi sinh vt a vo v mi trng nui thy sn. 3. Cc ch phm probiotic ch c tnh cht phng bnh. vic nui thy sn thnh cng, cn c nhiu gii php ng b. 4. Nh nc cn h tr cc c s nghin cu v sn xut trong nc nhanh chng t chc sn xut cc ch phm vi sinh s dng trong nui thy sn, cung cp vi gi phi chng, c nh vy mi m rng c din nhng ngi nui nui thy sn c p dng cc ch phm vi sinh vt.

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