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PHN I. LI M U Vit Nam c li th v din tch mt nc nui trng thy sn, tuy vy kh nng ng dng cha cao.

T trc nhng nm 1970, chng ta ch yu tp trung khai thc m cha thc s quan tm ti chnh sch pht trin v bo v ngun li t nhin. Nhng nm gn y cng vi chnh sch h tr ca nh nc, ngnh c nhng bc pht trin quan trng vi quy m ngy cng ln v din tch ngy cng c m rng. Trong xu th pht trin sn xut th nhu cu con ging ngy cng tng v cn c gii quyt. Ngy nay, cng vi s pht trin ca khoa hc k thut nhiu loi c kinh t, c bit l cc loi c bin c sn xut ging nhn to thnh cng. Cng ngh sn xut ging bao gm nhng gii php k thut quan trng, trong vn thc n v cch cho n khi ng u trng c l khu then cht quyt nh s tng trng v t l sng ca u trng trong sut qu trnh bin thi. To l thc n c gi tr dinh dng ln, c bit l protein (chim 50 60 % trng lng kh), tip l lipid, hirocacbonat, cc xit bo. c nhiu nghin cu v dinh dng cc i tng nui v mc d sn xut nhiu thc n nhn to cho u trng ca c, tm cng nh cc i tng nui hi sn khc, nhng khng c loi thc n no c th so snh c vi thc n t nhin. G.G.Vinbe (1965) nh gi vai tr ca to: Khng c to th khng c ngh c.ng vy to l mt xch u tin trong chui thc n mi trng bin, l thc n cn thit cho tt c cc giai on trong ng nui cc loi nhuyn th hai mnh v (Hu, Ngao, ip), giai on u trng ca cc loi ng vt chn bng v u trng ca mt s loi c bin v tm He hoc c th l thc n gin tip cho u trng cc loi trn thng qua zooplankton (lun trng, gip xc chn cho, artemia). Vi to cn c nui trc tip trong cc b ng nui u trng c bin trong k thut nc xanh vi vai tr lm n nh cht lng nc, dinh dng u trng v kim sot vi khun. V th nhn sinh khi to lun l cng on khng th thiu trong cc tri nui trng thy sn. Bn cnh nhng nghin cu v tm quan trng ca vi to th vic nghin cu cc k thut nui trng cho cht lng tt, nng sut sinh khi ln cng c nhiu ngi quan tm v ch pht trin. Nhm b sung kin thc thc t, kt hp l thuyt vi thc tin sn xut, ti c trng phn cng v trung tm chp nhn cho lm chuyn : Theo di quy trnh nui to lm thc n cho u trng c bin. Vi mc tiu l nm vng k thut gi ging, nui sinh khi, thu c nng sut cao trong sn xut v ch ng trong vic cung cp thc n phc v cho sn xut ging c bin. Chuyn gm c hai ni dung chnh + K thut lu gi ging to. + K thut nui sinh khi.

Vng Tu, thng 7 nm 2006 Sinh vin thc hin o th Hng PHN II. TNG QUAN TI LIU 2.1 S Lc V Tnh Hnh Nui V S Dng Mt S Loi Vi To Trn Th Gii V Vit Nam 2.1.1 Trn th gii

Vic nghin cu to c t lu i, n gn lin vi s ra i ca Knh hin vi quang hc v vic tm thy t bo u tin do nh t nhin hc ngi Anh R.Hooke vo nm 1665. Tuy nhin, mi n nm 1910, Allen v Nelson ln u tin nui to Silic lm thc n cho mt s ng vt khng xng sng (Ryther v Goldman, 1975). Nm 1939, Bruce v cng tc vin phn lp v nui to n bo Isochrysis galbana v Platymonas grossii nui u trng Hu. Nm 1940, Matsue pht trin phng php phn lp v nui thun Skeletonema costatum dng lm thc n cho u trng tm Penaeus japonicus giai on zoea t t l sng 30 % (thay v trc ch t 1 %) (Liao,1983). M Chlorella c bt u sn xut i tr nm 1950, (ng nh Kim, 1999). Nm 1953, cc nh khoa hc (Ty c) s dng kh thi CO2 ca nh my cng nghip vng Rubin nui trng to Chlorella sp v Scenedesmus acutus. Cng loi to ny, nm 1957, Tamiya v cng s vin sinh hc Tokugama (Tokyo) cng b kt qu nui trng to Chlorella ngoi tri v cng t Chlorella h chit ra mt hp cht gi l nhn t sinh trng Chlorella cng vi 15 loi Vitamin khc nhau, c ng dng rng ri trong y hc (V Hnh, 1996). Nm 1960, Scenedesmus c sn xut i tr Tip Khc, c, Israel, Italia v mt s nc ng u, ng thi to Lam Spirulina c trng i tr Mhico, M, i Loan, Israel, Trung Quc, Thi Lan, n v Vit Nam (ng nh Kim, 1999). Nm 1996 1997, Nht Bn sn xut to Chlorella t sn lng 1 100 tn/nm v Trung Quc sn xut 2 798 tn/nm to Spirulina xut khu (Yuan Kun Lee, 1997). 2.1.2 Vit Nam

u nhng nm 1960 gio s Nguyn Hu Thc v cc cng s tin hnh nhiu th nghim s dng sinh khi Chlorella vo thc n mt s loi gia cm, kch thch tng trng ca tm con (ng nh Kim, 1999). Nhng nm u 1970, sn xut ging Hi Sn bt u c quan tm, trm nghin cu nui trng thy sn nc l Qu Kim Hi Phng kt hp thu nui sinh khi to Silic lm thc n cho u trng tm mang tnh cht phc v sn xut l chnh ng thi tin hnh mt s th nghim nghin cu c bn. Nm 1974 mt s th nghim nui to Skeletonema costatum trong iu kin phng th nghim c tin hnh trng i hc Thy Sn Nha Trang. Cng loi to ny nm 1989 ti trung tm nghin cu thy sn III, Nguyn Th Xun Thu v cc cng tc vin tin hnh mt s th nghim v mi trng nui cy tm hiu kh nng pht trin ca Skeletonema costatum v Chaetoceros sp, s dng Skeletonema costatum ng u trng tm s giai on Zoea (L Vin Ch, 1996). Cng trong thi gian ny, lin doanh Vit Nam Oxtrylia v sn xut tm ging, s dng Skeletonema costatum lm thc n cho u trng t kt qu tt (ng nh Kim,2000). Nm 1996, L Vin Ch nghin cu mt s c im sinh hc, cng ngh nui to Silic Skeletonema costatum (Greville) Cleve lm thc n cho u trng tm bin. Trn Th Tho v cng s (2000) tin hnh nhng th nghim nui i tr Chlorella pyrenoidosa ti b 10 m3 i vi mi trng m v c c b sung hu c (0.4% bt c) v dch chit t (1%) (ng nh Kim, 2002).

2.2

Vai Tr Ca Thc Vt Ni Trong Nui Trng Thy sn

Mc d cc nghin cu v to c tin hnh mun hn cc lnh vc khc trong ngnh nui trng thy sn nhng vi nhng g ngin cu c cho thy to c vai tr ht sc quan trng trong thnh cng ca lnh vc sn xut ging v nui thng phm hu ht cc i tng nui trng thy sn. To l thc n l thc n trc tip cho u trng c, tm, ng vt thn mm hai mnh v. Cc nh khoa hc Vit Nam tin hnh th nghim a to Spirulina sinh khi vo thc n ca c M Trng, Trm C, R Phi vi t l 5% lm tng t l sng v tng trng ca c. To cn c dng sn xut khi lng ln cc ng vt ph du (lun trng, ng vt chn cho, artemia). Cc ng vt ph du ny li c dng lm thc n cho cc giai on u trng ca mt s loi tm, c. Ngoi ra, trong cc b ng u trng c bin, to cn c vai tr n nh cht lng nc v kim sot vi khun. Bn cnh vai tr lm thc n cho ng vt thy sn, to cn sn xut ra lng ln kh xi ho tan vo ban ngy (khi to quang hp) gp phn lm n nh nhit v pH trong mi trng ca cc ao, h khi tin hnh nui chung vi ng vt thy sn.

Mt s loi to nh Chlorella Nht Bn c ng vin v s dng nh mt loi thc n b dng cho ngi (ng nh Kim, 2000). S dng Chlorell, Scenedesmus, Spirulina vo khu phn thc n ca g vi t l 7,5 10% lm tng t l v hm lng vitamine A trong trng (ng nh Kim, 2000). Ngoi ra, to cn c sn xut i tr chit sut cc hot cht nh : Vitamine, lipid, cc sc t (carotenoit, chlorophyll( a,b,c1,c2), phycobiliprotein), cacbohydrat, mt s cht chng xi ho. Mt s loi to Lam c kh nng c nh m t Nit khng kh v c vai tr quan trng trong chu trnh bin i nit nn to Lam l loi phn bn sinh hc rt tt cho cy trng. To cng l phng tin lm sch nc thi c hiu qu vi gi thnh thp nht (ng nh Kim, 1999). Ni chung to c s lng loi rt ln, c nghin cu v ng dng rng ri trong dinh dng, cng nghip, y t, thy sn, nng nghip v bo v mi trng. Tuy vy, cc hng nghin cu sn xut i tr mt s loi to li ch yu phc v cho sn xut ging v nui ng vt thy sn.

Bng 1 Cc lp v ging vi to ch yu c nui trng dng cho nui thy sn ( sa i t ti liu ca De Pauw v Persoone, 1998) Lp Ging Bacillariophyceae Skeletonema Thalassiosira Phaeodactylum Chaetoceros Cylindrotheca Bellerochea Actinocyclus Nitzchia Cyclotella Haptophyceae Isochrysis Pseudoisochrysis Dicrateria Chrysophyceae Monochrysis (Pavlova) Prasinophyceae Tetraselmis (Platymonas) Pyramimonas Micromonas Nhng th d v p dng PL,BL,BP PL,BL,BP PL,BL,BP,ML,BS PL,BL,BP,BS PL BP BP BS BS PL,BL,BP,ML,BS BL,BP,ML BP BL,BP,BS,MR PL,BL,BP,AL,BS,MR BL,BP BP

2.3

Chroomonas BP Cryptomonas BP Rhodomonas BL,BP Cryptophyceae Chlamydomonas BL,BP,FZ,MR,BS Chlorococcum BP Xanthophyceae Olisthodiscus BP Chlorophyceae Carteria BP Dunaliella BP,BS,MR Cyanophyceae Spirulina PL,BP,BS,MR PL: u trng tm he Panaeid BL: u trng nhuyn th hai mnh v ML: u trng tm nc ngt BP: hu u trng nhuyn th hai mnh v AL: u trng bo ng; MR: lun trng bin (Brachionus) BS: tm ng mui (Artemia) SC: ng vt chn cho noc mn; FZ: ng vt ph du nc ngt. Gi Tr Dinh Dng Ca Vi To

Crytophyceae

To l ngun b sung dinh dng rt quan trng ca ng vt thy sn, l thc n khng th thiu tt c cc giai on ca cc loi nhuyn th hai mnh v. Mc d c nhng khc bit trong thnh phn sinh ho ca cc lp v cc loi to nhng Protein lun l thnh phn hu c ch yu (chim 12 35% trng lng kh ca vi to), tip l Lipid (7.2 23%) ri n Cacbohidrate (4.6 23%), (Brown, 1991). Hm lng cc axt bo khng no (HUFA), c bit l axt eicosapentaenoic (20:5n-3, EPA), axt arachidonic (20:4n-6, ARA) v axt docosahexaonic (22:6n-3, DHA) ng vai tr quan trng ch yu trong vic nh gi thnh phn dinh dng ca mt loi to dng lm thc n cho cc sinh vt bin. Nng quan trng ca axt eicosapentaenoic (EPA) u c mt cc loi to khu (Chaetoceros calcitraans, C.gracilic, S.costatum, T.pseudonana) v to Platymonas sp v to Chroomonas salina (Volkman v cs, 1989). Cc vi to cn c coi l ngun giu axt ascorbic (0.11 1.62% trng lng kh). Gi tr dinh dng ca to c th bin i ng k theo mi trng nui. Th d nh hng ca thnh phn ca mi trng nui ln thnh phn gn ng ca cc loi vi to khc nhau (Brown v Miller, 1992). Hm lng Protein trong mi t bo vn c xem lmt trong nhng yu t quan trng nht quyt nh gi tr dinh dng ca vi to dng lm thc n trong nui trng thy sn. Protein cng c chng minh l thnh phn nhy cm nht vi nhng bin ng ca mi trng nui hn nhng thnh phn cu to t bo khc.

Bng 2 Thnh phn sinh ha ca mt s loi to Loi to Spirulina platensis S. platensis S. maxima S. maxima Chlorella vulgaris C. pyrenoidosa Scenedesmus obliquus S. obliquus S. quadricauda Dunadiella salina D. bioculata Euglena gracilis Skeletonema sp Chaetoceros sp Nanochloropsis oculata Tetrselmis chui Isochrysis galbana Hm lng cc cht (% trng lng kh) Protein Lipid Carbohydrat 46 50 49 8 14 62.5 3 8.5 65 2 20 60 71 67 13 16 51 58 14 12 12 17 57 2 26 50 56 12 14 10 17 52 9 12.5 47 1.9 57 6 32 49 8 4 39 61 14 20 14 18 37 4.7 20.8 35 6.9 66 35 18 7.8 31 10 12 20 23 12.9

Tham kho Tipnis (1960) Becker (1984) Miller (1968) Duran (1980) Trubachev (1976) Aaroson (1980) Soeder (1981) Becker (1984) Hindak (1968) Parson (1961) Eddy (1956) Collyer (1955) Parson (1961) Parson (1961) Brow(1991) Brow(1991) Brow(1991)

Tuy nhin, tc sinh trng ca cc vt cho n bng hn hp cc loi to khc nhau thng cao hn cc vt ch c cho n bng mt loi to. Mt loi to c bit c th thiu mt cht dinh dng, trong khi mt loi to khc c th cha cht dinh dng v thiu cht dinh dng khc. Do hn hp c hai loi to s cung cp cho cc ng vt mt lng y gm c hai cht dinh dng. Vic xem xt chung cc kha cnh dinh dng ca cc vi to c s dng trong nui bin cc loi nhuyn th hai mnh v, gip xc v c c trnh by trong ti liu ca Brown v cc tc gi khc (1989). Liao, 1983 cho rng khng c loi to n c no li tt nht v mi phng din cho vic nui v s dng chng lm thc n cho ng vt thy sn, nhng hin nay phng php s dng ch mt loi to duy nht vn l phng php ch yu trong nui trng. 2.4 2.4.1 S Lc V c im Sinh Hc Ca Mt S Loi To Nui Chlorella

To Chlorella thuc ngnh to lc (Chlorophyta), l to n bo, t bo hnh cu hoc hnh van, khng c tin mao, khng c kh nng di ng ch ng, kch thc t bo t 5 7 m ty loi, ty iu kin nui v giai on pht trin ca to.

Chlorella phn b rng c th sng mn t: 5 30 , nhit : 15 35 0C, pH: 7.5 8.5. Ti ni c ngun cht thi hu c to pht trin t mt 50 x 10 6 tb/ml (ng nh Kim, 1999). Thnh phn ho hc ca Chlorella ph thuc vo s c mt ca Nit trong mi trng, khi i m hm lng prtin gim i r rt trong khi lng Cabohyrat v axt bo li tng ng k. y l thc n tt cho Brachionus plicatilis v u trng tm c. (Hnh 1). 2.4.2 Nannochloropsis oculata To Nanochloropsis oculata ging Chlorella nhng kch thc b hn. Nanochloropsis oculata pht trin tt trong mi trng dinh dng Walne (l mi trng c b sung vitamin B1 v B12), mn 18 26 , nhit 250C v mt nui cy ban u l 2 x 106 tb/ml (V Dng, 1998). Sinh trng v pht trin nhanh, trong iu kin ti u c th t 24,5 g/m2.ngy v nng sut Lipid l 4 g/m2.ngy (Boussiba etal, 1986). Trong iu kin nui tt, sau 7 ngy c th t mt 95 x 106 tb/ml. Nannochloropsis oculata l thc n thch hp cho nui lun trng Brachionus plicatilis. (Hnh 2) 2.4.3 Tetraselmis chui (= Platymonas sp)

To Tetraselmis chui c t bo hnh dt, c 4 tin mao, di ng mnh trong nc, sinh trng chm hn Nannochloropsis oculata. Mi trng tt nht cho s pht trin ca Tetraselmis chui: mn 30, nhit 30 C (V Dng, 1998), Tetraselmis chui pht trin tt trong mi trng dinh dng Walne, Guillard v Ryther. Thnh phn axt bo ca to ny c c trng bi t l axit bo khng no cao c bit l C18:3 (18.9 %), C18:4 (7.5 %), C20:5 (7.5 %), v C22:6 (1.4 %) (Pohl v cng s, 1979).
0

Tetraselmis chui l thc n thch hp cho u trng hai mnh v (Pe Pauw, 1983) v c s dng rng ri trong lm thc n cho u trng tm, bo ng, Artemia v lun trng. (Hnh 3). 2.4.4 7 m. T bo cha 7,07% khi lng tnh l lipid, t l axt bo khng no chim 60% tng lng xt bo, Isochrysis galbana l mt trong nhng loi vi to bin tch lu trong c th lng cht bo rt cao, i khi chim ti 40% trng lng kh. Isochrysis galbana l thc n tt cho zooplankton (u trng thn mm hai mnh v, copepoda v lun trng) (Toonen, 1995). (Hnh 4) 2.5 Mt S Yu T Mi Trng nh Hng Ti Sinh Trng Ca To Nui 7 Isochrysis galbana To Isochrysis galbana thuc lp Chrysophyceae. C mu nu vng, kch thc t 5

2.5.1

Nhit

Nhit ao h nui bin ng theo ngy m (cao nht vo lc 14h v thp nht vo 5 6h), theo ma v theo su, nhng ao h c din tch cng nh th bin dao ng nhit cng ln, nhng ao h nh nh sng mt tri nhanh t nng lp nc b mt lm t trng nc tng ny nh hn tng di, do m kh nng xo trn gia cc tng nc km lm nc phn tng r rt. Nhit nh hng rt ln n thc vt thy sinh c bit l phytoplankton, khi nhit cao qu hoc thp qu c th lm tng hoc gim qu trnh trao i cht gy ri lon sinh l sinh vt. Nhit m a s cc loi to c kh nng chu ng l 16 35 0C, nu nhit thp hn 16 0C th to pht trin km, nhng nu cao hn 35 0C th to s cht. Nhit ti u cho a s cc loi to l 20 24 0C, tuy nhin gi tr ny bin i tu loi. 2.5.2 nh sng

Vi to l loi quang t dng, chng s dng nng lng nh sng mt tri, dng cht v cc khong vi lng t tng hp cht hu c cho c th nn thi gian chiu sng v cng nh sng nh hng rt ln n s pht trin ca to nui. Tuy nhin cng chiu sng v thi gian chiu sng ph thuc vo tng loi to, mc d chng sng v pht trin l nh qu trnh tng hp cht hu c di iu kin nh sng mt tri nhng thi gian chiu sng ti a cha hn l tt nht, v d nh to Skeletonema costatum th thi gian chiu sng 12h/24h (theo chu k ngy m) l tt nht (L Vin Ch, 1996), cng chiu sng cng ph thuc vo th tch nui, th tch nh th cng chiu sng c th nh nhng nu th tch nui ln th cng chiu sng phi ln kt hp vi s c kh v o nc. 2.5.3 pH

pH c nh hng rt ln n i sng ca thy sinh vt, mi loi sinh vt ch tn li trong mt khong pH nht nh, nu tng qu hoc gim qu s gy ri lon trao i cht ca sinh vt. c bit l i vi thc vt thy sinh chng s dng CO 2 cho qu trnh tng hp cht hu c, lm dch chuyn h cn bng bicacbonat: 2HCO3 - CO32- + CO2 + H2O (1) CO32- + H2O HCO3 - + OH- (2) Khi to quang hp phng trnh (1) v (2) tch cc chuyn sang pha phi chng li s gim CO2 kt qu lm nc tch nhiu ion OH- lm pH tng cao, khi to t mt cc i pH c th 9. pH trong b nui cn c tc dng kim sot H2S. nhit 24 0C:

+ Nu pH = 5 th 99,1% H2S dng kh c. + Nu pH = 9 th ch c 1% H2S dng kh c. 2.5.4 mn

i vi cc thc vt ni nh vi to th mui nh hng rt ln n sinh trng v pht trin, iu ny th hin khi ma ln hoc nng ln ko di c th dn n s thay i thnh phn v s lng cc loi to trong cc thy vc t nhin, ngoi ra n cn nh hng ti s pht trin ca ng vt ph du (zooplankton). 2.5.5 Sc kh

Sc kh l khu rt quan trng khng th thiu trong nui vi to nht l nhng b nui c su ln. Sc kh c tc dng o u to (rt c ngha i vi nhng to c kch thc t bo ln, d lng) gip cc t bo tip xc u vi nh sng, dinh dng, gim hin tng phn tng nhit v cn c tc dng ln trong vic cung cp CO 2 cho quang hp ca to. Sc kh phi m bo 24h/24h. 2.5.6 nh hng ca mt ging ban u ln s pht trin ca to

Trong cc yu t nh hng ti sinh trng qun th to, mt ging ban u cho chng ta kh nng tc ng tch cc nht. Trong cng mt iu kin mi trng, mt ging cao s to kh nng sinh trng nhanh v t mt cc i cao. Mt nui ban u m tha th nh sng d phn b u hn trong cng mt th tch nui, nhng nu mt ban u m tha qu th n nh hng n cnh tranh pht trin ca vi khun. V d v to Sketonema cotatum mt ban u l 2 x 104 tb/ml, phi sau 54 60h to mi t mt cc i, nhng vi mt ban u l 10 x 104 tb/ml th ch sau 30 36h to t mt cc i ( L Ch Vin,1996). Tuy nhin, lng ging to s dng mc no l do mc ch ca ngi s dng. Trong nhn sinh khi to lm thc n cho ng vt nui, ngi ta quan tm nht l khi lng sinh khi to t c trong thi gian nht nh nn mt to ging ban u thng ln. 2.6 ng Lc Hc Sinh Trng th1 tng trng chung ca mt s loi to

S tng trng ca cc vi to trong iu kin v trng c c trng bi 4 pha + Pha chm hoc pha cm ng

Pha ny, mt t bo tng t hoc c th khng tng, giai on ny tng i di khi chuyn t mi trng thch sang mi trng lng. Vic chm pht trin l do s thch nghi sinh l ca s chuyn ho t bo pht trin, nhng tng cc mc enzym v cc cht chuyn ho lin quan n s phn chia t bo v s chuyn ho ccbon. + Pha sinh trng theo hm s m Trong pha ny mt to tng nh hm s m ca thi gian t theo hm lgarit: Ct = Co.emt Vi Ct v Co l cc nng t bo ti thi im t v 0 tng ng m m= tc sinh trng c th ph thuc ch yu vo loi to, cng nh sng v nhit . + Pha dng pha ny yu t hn ch v tc sinh trng c cn bng, s t bo sinh ra tng ng vi s t bo cht i, lm mt to khng tng hoc tng rt t. + Pha tn li Trong pha cui cng ny, cht lng nc xu i v cc cht dinh dng suy kit ti mc khng th duy tr c s sinh trng. Mt t bo gim nhanh v qu trnh nui cng kt thc ti y. 2.7 2.7.1 Mt S Phng Php Nui To c im c bn ca mt s phng php nui to Bng 3 c im ca cc phng php nui to Phng php Nui trong phng c im u im Nhc im

To c nui trong Cc yu t mi trng u t k thut cao phng th nghim hay nh nhit , mn, v t tin nh kn nh sng c khng ch thch hp. Hn ch c tc ng bn ngoi, hn ch nhim tp, nhim khun. Quy trnh k thut c m bo. Sn xut ch ng To c nui trc R, n gin, c th tip ngoi tri, s nui vi quy m ln. dng nc bin, nh Kh kim sot mi trng m hon ton ph thuc vo thi

Nui ngoi tri

10

sng t nhin.

tit, kh hu. Khi iu kin mi trng bin i khc nghit s lm to suy tn v cht, kh khc phc. To nui d nhim tp. Kh ch ng trong sn xut. t nhim tp (to tp, vi Quy m nh. t khun, protozoa hay tin. Thao tc nui rotifer). Ch ng cy cn thn. trong sn xut. D nhim tp, nh hng n to nui.

Nui kn

To c nui trong cc dng c kn nh ng nghim, bnh tam gic, ti nha.

Nui h

Nui chuyn

To c nui ngoi R. Quy m ln. tri hay trong phng, nui trong nhng ao hay b, khng y kn. D thc hin nht. Ph Nui trong phng hay bin nht v ng tin ngoi tri. To b cy nht. th tch nh khi t mt cao s c lm ging nui cy vo b c th tch ln hn v c th. b sn xut, to mt cao s c thu hoch.

Tn nhiu cng, t kinh t. Cht lng to cc b ln c th khng m bo.

2.7.2

K thut nui to theo phng php nui chuyn

C nhiu phng php nui to: Nui chuyn, nui lin tc, nui bn lin tc ty vo iu kin c th m la chn phng php nui ph hp. Do tnh linh hot v d khc phc nhng sai st trong h thng nui nn bin php ch yu trong phng th nghim l phng php nui chuyn. c im ca phng php ny l khi to t mt cao s c lm ging nui cy vo th tch ln hn. Trong phng php ny cn cc dng c nui c nhiu th tch t 250 ml 2000 lt. To c lu gi trong cc ng nghim cha 20 ml dung dch dinh dng hoc trn mi trng thch. i vi to roi cn chiu sng lin tc, ring to khu ch cn 12h/24h. Cng chiu sng cho giai on ny l 750 1000 lux (tng ng hai bng n Neon 40W), nhit 250C. giai on ny khng cn sc kh. i vi mi trng lng trong ng nghim

11

phi tin hnh kim tra v lc hng ngy bng my hoc lc bng tay. Tt c cc khu giai on ny u tuyt i v trng, trnh nhim khun v nhim tp. To c lu gi khong mt thng sau tin hnh cy sang mi vo bnh tam gic hoc sang ng nghim khc. Ly 2ml to gc t ng nghim cy vo bnh Erlen 250 ml cha 100 ml dung dch dinh dng, y kn bng nt cao su, sc kh nh, cng chiu sng 1000 1500 lux. Sau thi gian cy to trong bnh Erlen 250 ml t 3 6 ngy (to ang pht trin pha tng trng), tin hnh cy to qua cc bnh khc c th tch ln hn. Th tch to ging trong cc bnh chim 2 10% th tch nc nui mi. Cc bnh, keo c t trn cc gi nui c b tr 4 n Neon, c sc kh, nhit c nh 25 0C, mn nc nui cng c c nh 25, mi trng Walne c s dng giai on ny l 1 ml/l nc nui, cho mt ln khi mi cy. Cc bnh, keo c y kn bng nt cao su hoc np nha v tt c cc dng c nui c sy kh trng. Cc ti Nilon cng c s dng trong giai on ny. Thi gian nui cy giai on ny c th ko di 2 6 ngy song thng tin hnh sang mi sau 2 3 ngy nui. giai on sn xut to t cc keo hay bnh ln s c nui cy vo cc thng nha c th tch 100 lt. Sau chuyn dn ti b 500 lt, 1m 3, 2m3 v 60 m3. Th tch to ging giai on ny thng bng 10% th tch nc nui mi, mi trng dinh dng cho to lc ny a dng: mi trng Walne, Guillard F 2, Liao hay phn v c (phn bn nng nghip), ring mi trng Walne s dng vi liu lng 1 2 ml/10 lt nc nui. giai on ny cn sc kh mnh cn cc yu t khc (nh sng, nhit , mn) ph thuc hon ton vo t nhin. Thi gian nui giai on ny thng ngn, c th mt ngy hay 5 7 ngy nhng giai on ny thi gian nui di d b nhim tp. PHN III NI DUNG VT LIU V PHNG PHP NGHIN CU 3.1 3.1.1 Ni Dung Nghin Cu Thi gian v a im nghin cu Thi gian nghin cu t thng 5 8 nm 2006. a im: Trung Tm Quc Gia Ging Hi Sn Nam B Vin Nghin Cu Nui Trng Thy Sn II. 3.1.2 Ni dung nghin cu Theo di k thut lu gi ging to. Theo di k thut nhn ging v nui sinh khi to.

12

3.2 3.2.1

Vt Liu Nghin Cu To ging

i tng l cc loi to Nanochloropsis oculata, Tetraselmis chui (=Platymonas sp), Isochrysis galbana v Chlorella sp c ly t phng th nghim ca trung tm. 3.2.2 Mi trng nui to

C nhiu mi trng dinh dng dng nui to nh : Mi trng Walne, Guillard, Ryther, Tamya, Ito v mi trng phn v c. Nhng trong phm vi sn xut, c s s dng hai loi mi trng chnh l mi trng Walne (dng trong nhn ging v nhn sinh khi ngoi tri vi cc th tch nh) v mi trng phn v c (dng trong sn xut sinh khi to b ximng). 3.2.2.1 Thnh phn v cch pha ch mi trng Walne Bng 4 Thnh phn v cch pha ch mi trng Walne (da vo ti liu ca Laing, 1991) Thnh phn Dung dch A FeCl3 MnCl2 .4H2O H3BO3 EDTA NaH2PO4.2H2O/ KH2PO4.2H2O NaNO3/ KNO3 Tt c pha n 1 lt nc ngt Dung dch B ZnCl2 CoCl2.6H2O (NH4)6Mo7O24.4H2O CuSO4.5H2O Tt c pha n 100 ml nc ngt Dung dch C Vitamine B1 Vitamine B12 Tt c pha n 100 ml nc ngt + Cch pha mi trng - Cho 100 ml nc ngt vo bnh Erlen 2 lt, un si ri cho dung dch A vo theo th t nh s t 1 6 ( bng trn), ch trong qu trnh cho cc cht vo bnh kt hp khuy mnh, khi thy tan ht mi cho cc cht tip theo vo. - Cho dung dch B vo dung dch A vi liu lng 1 ml/1 lt dung dch A. (4) (5) (3) (1) (6) (2) S lng (g) 0.8 0.4 33.6 45 20 100 2.1 2.0 0.9 2.0 200mg 10mg

13

- Hp tit trng 121 0C, 1 atm v trong 20 pht. - nhit dung dch trn xung cn khong 50 60 0C, trn tip dung dch C vo dung dch A vi s lng vitamin (B1:1 ng/ 1 lt dung dch A, B12: 5 ng/1 lt dung dch A). - Cui cng ta c dung dch A tng hp dng bn cho to. Liu lng dng phng th nghim l 1 ml/1 lt to nui, nui ngoi tri dng 1 2 ml/10 lt to nui. 3.2.2.2 Thnh phn mi trng phn v c Bng 5 Thnh phn cc cht trong mi trng dinh dng phn v c Thnh phn SA (Sunfat Amonium) Ur Ln 3.2.3 Nc nui Nc dng trong nui to l nc bin x l theo quy trnh Nc bin Chlorine (30 ppm) b cha Thiosunfate (20 30 ppm) b lng lc ct 2 lc ct 1 UV lc ozone ti lc (50 m) b nui b x l Khi lng (g/m3) 100 10 15

Nc t bin vo b x l c x l bng Chlorine 30 ppm, sau 24h trung ho lng Chlorine d bng Natri Thiosulphat 30 ppm. Nc c chy qua mt h thng lc tinh trc khi cung cp cho cc b nui to. Nc dng gi ging v nui sinh khi trong phng th nghim phi h mn xung cn 25, hp tit trng 1210C, 1 atm trong 20 pht, sau ngui nc nhit phng nui. 3.2.4 Mi trng agar

Pha mi trng agar 1.5% (pha 1.5 gam agar trong 100 ml nc), un ho tan agar trn ngn la Bunsen khi thy agar tan ht em hp tit trng 121 0C, 1 atm trong 20 pht, nhit xung cn 50 60 0C, cho Vitamine vo lc u ri ra a Petri hoc ng nghim ( kh trng) vi th tch agar bng 1/3 th tch a Petri v 20 ml i vi ng nghim. t ng nghim nm nghing sao cho din tch b mt agar l ln nht. Sau ngui agar trong t cy v trng. 3.2.5 Cc thit b v dng c

14

3.2.5.1

Thit b v my mc + T gi to c iu kin: nh sng 3 000 lux, nhit : 250C, m 60 70%. + My sy dng c Drying oven MOV 212 + My hp nc Hiclave HVE 50

+ Mt s dng c khc nh : Knh hin vi quang hc, bung m hng cu, my o pH Themov orinon Model 280A, my o Ammonia DR2010, t cy v trng, n cn, khc x k + Gin nui to c b tr cc bng n Neon 40W, cng chiu sng t 6 000 10 000 lux, h thng cc ng v dy sc kh. + H thng sc kh bao gm my bm, cc ng nha PVC, cc dy kh v b lc kh.

u kh vo

u kh ra bng than ci hot tnh bng

S 1 Cu to b lc sc kh 3.2.5.2 Cc dng c nui to

Cc bnh Erlen th tch t 0.25 2 lt; cc bnh hnh tr c th tch t 3 10 lt; ti nilon; thng nha 100 lt ; cc b Composite c th tch t 0.5 2 m 3; b ximng 60 m3; cc ho cht v dng c khc. 3.3 3.3.1 Phng Php Nghin Cu Phng php lu gi ging to

Gi ging l bin php bo qun k thut nhm gi ging gc ch ng trong qu trnh sn xut. Vic chn to ging gi v nui sinh khi rt quan trng trc ht (l phi xt n tnh n ca ng vt ph du (lun trng, artemia, copepoda), tip l nng sut nui trng, thnh phn dinh dng ca vi to, kh nng chng chu tt vi iu kin ngoi cnh, tnh hnh phn b, khng cha c t, d tiu ho Yu cu gi ging: ging sch khng nhim tp, v th tt c cc thao tc k thut trong cy v gi ging to u phi m bo v trng. Trong sn xut ngi ta thng gi li hai lot ging gc, mt lot dng cho nui cy ban u phc v cc h thng sn xut (gi 15

ging trong mi trng lng), cn lot kia ch c x l khi cn lu gi ging (gi ging trong mi trng thch). To sau khi cy c gi iu kin nh sng yu cng chiu sng 3 000 lux (nh sng nhn to bng n Neon), nhit 25 0C, m 60 70 %. Sau 2 3 ngy s thy to pht trin, khong 4 5 ngy thy c mu c trng ca to. Hng ngy kim tra s pht trin ca to, nu thy nhim khun th phi loi b ngay trnh s ly nhim. Thi gian gi to l 8 tun, thng sau 4 tun tin hnh nui cy mi. To ging: Gm cc loi to Nannochloropsis oculata, Chlorella sp, Isochrysis galbana v Tetrselmis chui. 3.3.1.1 Gi ging trn mi trng thch a/ Chun b + ng nghim, a petri c sy kh trng + Mi trng nui to + Pha mi trng agar + Hp tit trng dung dch agar v bng cy. + mi trng thch. Mi trng thch sau khi hp tit trng ngui khong 50 60 C cho Vitamine vo, lc u ri vo ng nghim hoc a petri ( sy kh trng) sao cho lng agar trn mt a chim khong 1/3 th tch a petri hoc cho 20 ml agar vo ng nghim ri t ng nghim nm nghing sao cho din tch b mt agar l ln nht.
0

b/

Cy to

Chn ng nghim hoc a thch no c dy to ln p gi ging, dng bng cy (chn im no to pht trin p nht ly) Cy to theo hnh zic zc. Kh trng ming ca ng nghim v a petri bng ngn la n cn. Gin kn ming ng hoc a bng giy parapin (trnh nhim tp trong qu trnh gi ging). To t mi trng lng trong ng nghim c cho vo falcon 15 ml ly tm 3000 vng/pht trong 1 pht (to s lng phn y), b lp nc trn i, phn cn li cho vo my lc, lc u ri dng bng cy ly to, cc thao tc tng t trn. 3.3.1.2 Gi ging trong mi trng lng a/ Mi trng nui 16

+ Nc bin x l + Mi trng dinh dng Walne (1 ml/l nc nui). b/ Phng php cy to t mi trng thch sang mi trng lng

Chn trn a thch c to pht trin tt dng que cy trn ly to ging t a thch (chn ng cy c to pht trin tt nht) v cho vo ng nghim ng 10 ml nc bin c cha mi trng dinh dng, lc u bng my lc, y kn np ng nghim v gin kn bng giy parapin. c/ Phng php nui + Gi ging trong t nui to c iu kin 250C, m 60 700C, nh sng 3000 lux. + Hng ngy tin hnh lc to bng my lc + Sau 2 3 ngy s thy to pht trin. 3.3.2 Nhn sinh khi mt s loi to

3.3.2.1 Nhn sinh khi trong phng th nghim a/ Chun b + Dng c - Cc bnh nui to trong phng th nghim c th tch t 0.1 10 lt, ti nilong. - Kh trng bnh nui: Trong nui to, c bit l nui cy ging trong phng th nghim th tt c cc khu u phi v trng (trnh nhim tp). Cc bnh nui to sau khi nui phi c ra sch bng x bng, sau ra k li bng nc ngt, bnh kh ri sy kh trng iu kin 170 0C, 1atm trong 2 gi. + Nc nui to Nc nui to phng th nghim l nc bin c x l, c mn 25 0/00, c hp tit trng 121 0C, 1 atm trong 20 pht. Sau ngui nc nhit phng. b/ Cy to

Ly 10 20 ml to gc t ng nghim cy vo bnh tam gic 100 ml cha 50 80 ml dung dch dinh dng Walne.

17

y bnh bng nt cao su c l thng kh. Sau thi gian 3 6 ngy khi to pha logarit tin hnh cy sang th tch mi, cc giai on ny to ging dng vi t l 10 20 % th tch nui mi. c/
0

Phng php nui to

Cc bnh nui trong phng th nghim c cc iu kin tng i n nh. Nhit 25 C, nh sng v sc kh 24/24 gi. cc th tch nh hn mt lt sc kh nh v nh sng yu (2 bng n Neon 40W). i vi cc th tch ln hn hai lt b tr sc kh mnh, cng nh sng ln (4 bng n Neon 40W). 3.3.2.2 Nhn sinh khi ngoi tri a/ Chun b + Cc dng c nui to - Cc thng c th tch 100 lt, cc b Composite c th tch t 0.5 2 m 3 v cc b xi mng c th tch 60 m3. - Cc dng c sau khi kt thc 1 t nui phi tin hnh kh trng bng x bng, sau ra k li bng nc ngt (i vi cc loi to kch thc nh nh Nanochloropsis oculata v Chlorella sp th phi kh trng b bng Chlorine 3 5 ppm). + Nc nui to l nc bin x l. b/ Cy to

p dng phng php nui m (trong qu trnh nui khng thm bt k cht g vo mi trng nui to). + To ging c ly t phng th nghim. + Mt cy ban u ty loi, thng th tch ging bng 2 10 % th tch nc nui ban u. + Cy nhn ging t th tch nh ti th tch ln, sau 2 3 ngy khi to ang pha tng trng th tin hnh cy sang mi. c/ Phng php nui

H thng sc kh mnh, u hot ng lin tc 24/24 gi. Anh sng, nhit ph thuc vo iu kin t nhin, cc b nui to c b tr ngoi tri khng c mi che. Hng ngy kim tra sc kh, kim tra mc nhim tp ca to di knh hin vi. To ging ng nghim 18 Bnh Erlen Bnh hnh tr

t agar B ximng (60 m3) B composite (0.5 2 m3)

(0.1 2 lt) Thng nha (100 lt)

(5-10 lt)

Ti nilon

S 2 Phng php nhn ging t th tch nh ti th tch ln 3.3.3 Phng php theo di tc pht trin ca to

3.3.3.1 B tr th nghim + i tng: Nanochloropsis oculata, Tetrselmis chui, Isochrysis galbana, Chlorella sp. + Th tch bnh nui: tt c nui th tch 2 lt + Mi loi b tr ba t, mi t hai bnh + Cc iu kin nui tng i n nh: nhit 25 0C, mn 25, cng chiu sng t 6 000 10 000 lux, chiu sng v sc kh 24/24h. + Mt ging ban u tu loi: Nanochloropsis oculata: 10 11 triu tb/ml Platymonas sp: 2 3 triu tb/ml Isochrysis galbana: 8 9 triu tb/ml Chlorella sp: 6 7 triu tb/ml 3.3.1.2 Phng php theo di tc pht trin ca to + Hng ngy m mt to. + Dng bung m hng cu m s lng t bo to trc tip di knh hin vi nhm xc nh c thi gian pht trin ca cc pha. + Tng kt s liu v v th tng trng ca cc loi to. + Cu to ca bung m hng cu - Mi bung m c cc Block: A, B, C, D, v E - Mi Block: A, B, C, D c chia lm 16 vung nh, mi vung c din tch 1 mm2, su 0.1 m nn th tch ca mi s l 0.1 mm3 Nu m to trong mi 4 ny l X th mt to l:

19

d(tb/ml) = X x 104 (tb/ml) - Ring Block E c chia lm 25 vung nh, mi c th tch l 1/25 x 0.1 mm 3 (0.004 mm3 = 0.000004 ml = 4 x 10-6 ml), mi vung nh li c chia lm 16 vung nh hn. Nu m to trong mi nh (trong 25 vung) l X th mt to l: d(tb/ml) = X/V = X/4 x 106 (tb/ml) Trong : d : mt t bo (tb/ml). X: s t bo trung bnh ca 1 (bng tng s t bo m c/s m). V: th tch ca m. 3.3.4 Theo di s bin ng ca mt s yu t mi trng

3.3.4.1 Bin ng pH S dng my o pH Themor orinon. Model 280 A o. i vi cc l th nghim th o vo cc pha: pha chm (1), pha tng trng (2) v pha dng (3). ngoi cc b nui sinh khi, o pH lc mi cy to v lc thu hoch (cp to vo b nui lun trng hoc b c ng). 3.3.4.2 Amonia (NH3 N)

o Amonia bng phng php Idophenol. i vi cc l th nghim th o vo cc pha: pha chm, pha tng trng v pha dng. ngoi cc b nui sinh khi, o Amonia lc mi cy to v lc thu hoch. 3.3.4.3 Nhit - o nhit 2 ln/ngy vo 6h v 14h mi ngy. - S dng nhit k thy tinh o. 3.3.3.4 mn S dng khc x k o mn hng ngy. 3.3.5 Cc ch tiu theo di + Theo di s pht trin ca mt s loi to nui.

20

+ Theo di nh hng ca yu t nhit v mn ln s pht trin ca cc loi to nui. + Theo di bin ng hm lng Amonia v pH trong qu trnh pht trin ca cc loi to nui. 3.3.6 Phng php x l s liu S dng phn mm Excel x l s liu.

21

PHN IV. KT QU NGHIN CU 4.1 4.1.1 Theo Di Quy Trnh Lu Gi Ging Trong Phng Th Nghim Lu gi ging trn mi trng thch Bng 6 Kt qu lu gi ging trn mi trng thch Loi to S mu Bt u mc (ngy) 2.50.5 31.0 2.51.0 20.5 Mc u (ngy) 4.50.5 61.0 51.5 40.5 Mc dy (ngy) 80.5 91.5 82.0 61 Thi gian lu gi ngy 325.0 38.57.5 364.0 273.0

N. oculata T. chui I. galbana C. vulgaris

6 6 6 6

+ Qua thi gian theo di cho thy trong iu kin t gi to (nhit 25 0C, m 60 70 %, cng nh sng 300 lux) th a s cc loi to u pht trin tt. Sau khi cy 2 3 ngy to bt u mc (ty loi), sau 4 6 ngy to mc iu, c mu sc c trng ca ging, t ngy th 6 8 tr i to mc dy, mu m dn. + Sau 3 4 tun th tin hnh cy nhn ging mi trng lng hoc cy gi ging li trn mi trng thch. + C th gi ging to trn mi trng thch 7 8 tun. + Trong cc mu theo di th ti thy a s pht trin nhanh, u, mu sc p, nhng c 1 mu ca to I. galbana c b nhim to Lc trong qu trnh gi ging. + Kt qu thnh cng (trong cc t theo di) t t l kh cao 95,8 %. 4.1.2 Lu gi ging trn mi trng lng Bng 7 Kt qu lu gi ging trn mi trng lng Loi to S mu Bt u mc (ngy) 1.50.5 2.50.5 2.00.5 Mc u (ngy) 4.00.5 4.50.5 4.50.5 Mc dy (ngy) 6.00.5 7.50.5 7.00.5 Thi gian lu gi ngy 15.03.0 22.04.5 162.5

N. oculata T. chui I. galbana

6 6 6

C. vulgaris 6 1.50.5 4.00.0 6.00.5 163.0 + To ging mi trng lng sau 1 3 ngy bt u mc, sau 4 5 ngy to mc iu, c mu sc c trng ca ging, t ngy th 6 8 tr i to mc dy, mu m dn. 22

+ Trong qu trnh theo di th cng c 1 mu I. galbana mi trng lng b nhim to Lc. 4.2 4.2.1 Theo Di Quy Trnh Nhn Sinh Khi Mt S Loi To Nhn sinh khi trong phng th nghim

4.2.1.1Nhn sinh khi to th tch 0.5 lt Bng 8 Mt to nui th tch 0.5 lt Loi to S mu Mt (x 106 tb/ml) Mi cy 7.4 0.53 1.920.11 5.70.36 4.70.26 Thu hoch 27.8 1.71 5.930.59 20.72.71 20.81.39 Thi gian nui (ngy) 3 3 3 3 % tng trng 275.7 208.9 263.2 342.6

N. oculata T. chui I. galbana C. vulgaris

3 3 3 3

+ th tch 0.5 lt cc loi to pht trin tng i nhanh, sau 3 ngy nui to N. oculata tng 275.7 % so vi ging ban u T. chui tng 208.9 % so vi ging ban u I. galbana tng 263.2% so vi ging ban u C. vulgaris tng 342.6 % so vi ging ban u 4.2.1.2Nhn sinh khi to th tch 1 lt Bng 9 Mt to nui th tch 1 lt Loi to S mu Mt (x 106 tb/ml) Thi gian nui (ngy) 3 3 3 3 % tng trng 249.5 187.8 228.6 370.3

Mi cy Thu hoch N. oculata 3 7.230.25 25.271.50 T. chui 3 2.210.17 6.360.31 I. galbana 3 4.20.26 13.82.15 C. vulgaris 3 9.10.57 42.80.85 + th tch 1 lt, sau 3 ngy nui to N. oculata tng 249.5 % so vi ging ban u T. chui tng 187.8 % so vi ging ban u I. galbana tng 228.6 % so vi ging ban u C. vulgaris tng 370.3 % so vi ging ban u 4.2.1.3Nhn sinh khi to th tch 2 lt 23

Bng 10 Mt to nui th tch 2 lt Loi to S mu Mt (x 106 tb/ml) Mi cy N. oculata T. chui I. galbana C. vulgaris 3 3 3 3 10.61.2 3.090.42 8.40.36 7.40.2 Thu hoch 24.03.7 6.670.15 18.00.42 17.30.9 Thi gian nui (ngy) 2 2 2 2 % tng trng 126.4 115.9 114.3 133.8

th tch 2 lt, sau 2 ngy nui to N. oculata tng 126.4 % so vi ging ban u T. chui tng 115.9 % so vi ging ban u I. galbana tng 114.3 % so vi ging ban u C. vulgaris tng 133.8 % so vi ging ban u 4.2.1.4 Nhn sinh khi to ti nilon Bng 11 Mt to nui ti nilon Loi to S mu Mt (x 106 tb/ml) Mi cy 1.130.1 7.61.1 Thu hoch 3.70.8 25.32.5 Thi gian nui (ngy) 3.50.5 3.50.5 % tng trng

T. chui N.Oculta

3 3

236.4 232.9

+ Nui ti nilon l mt bc chuyn tip nhn ging cc th tch ln ngoi tri. + Vi mt ging ban u 1.130.1 (x 106 tb/ml) i vi T. chui v 7.61.1 (x 106 tb/ml) i vi N.Oculta, sau 3 4 ngy nui mt T. chui t 3.70.8 (x 106 tb/ml) tng trng 236.4 % so vi ging ban u, mt N.Oculta sau 3 4 ngy nui t 25.32.5 (x 106 tb/ml) tng trng 232.9 % so vi ging ban u.

24

Qua cc loi theo di tt c cc th tch cho thy: + cng 1 th tch to C. vulgaris pht trin nhanh nht, tip l N. oculata v T .chui cui cng l I. galbana pht trin chm nht. + Cng 1 loi to th tch nh to pht trin nhanh hn cc th tch ln. V d: to N. oculata th tch 0.5 lt tc tng trng ca to t 275.7 % so vi ging ban u 1 lt tc tng trng ca to t 246.7 % so vi ging ban u 2 lt tc tng trng ca to t 126.4 % so vi ging ban u 4.2.2 Nhn sinh khi cc th tch ngoi tri

4.2.2.1Nhn sinh khi to th tch 100 lt

Bng 12 Mt to nui th tch 0.1 m3 Loi to S mu Mt (x 106 tb/ml) Mi cy N. oculata T. chui I. galbana C. vulgaris 3 3 3 3 2.50.66 0.370.05 0.90.23 2.30.21 Thu hoch 5.60.96 0.870.12 1.80.31 5.70.25 2 2 2 2 108.0 135.1 100.0 147.8 Thi gian nui (ngy) % tng trng

+ Thi gian nui 2 ngy + Tc tng trng ca N. oculata t 108.0 %, T. chui t 135.1 %, I. galbana t 100.0 % v C. vulgaris t 147.8 % 4.2.2.2Nhn sinh khi to th tch 500 lt Bng 13 Mt to nui th tch 0.5 m3 Loi to S mu Mt (x 106 tb/ml) Mi cy T. chui I. galbana C. vulgaris 3 3 3 0.230.03 1.30.5 2.00.25 Thu hoch 0.700.07 2.40.45 4.80.70 2 1 2 204.3 84.6 140.0 Thi gian nui (ngy) % tng trng

25

+ Thi gian nui 1 2 ngy + Tc tng trng ca N. oculata t 108.0 %, T. chui t 204.4 %, I. galbana t 100.0 % v C. vulgaris t 147.8 %. + Sau 1 2 ngy nui thu hoch to lm ging nhn mi cc th tch 0.5 m3 v 1 3 m . Ring to N. oculata kt thc nui sinh khi th tch 0.1 m3 do nhit qu cao, to pht trin km, thay vo to T. chui nui nhiu t th tch 0.1 m3 n th tch b ximng. 4.2.2.3Nhn sinh khi to th tch 1m3 Bng 14 Mt to nui th tch 1 m3 Loito S mu Mt (x 106 tb/ml) Mi cy T. chui I. galbana C. vulgaris 3 3 3 0.250.04 1.200.1 2.130.15 Thu hoch 0.710.06 3.170.15 5.870.15 2 2 2 184.0 164.2 175.6 Thi gian nui (ngy) % tng trng

+ Thi gian nui 2 ngy + Tc tng trng T. chui t 184.0 % sau 2 ngy nui, I. galbana t 8v t 171.4 % to C. vulgaris. + To I. galbana ch nhn n th tch 1 m3 l chuyn nui ra b ximng. + Sau 2 ngy thu hoch mt phn dng cho lun trng n l mt phn dng lm ging nhn mi. 4.2.2.3Nhn sinh khi to th tch 2m3 Bng 15 Mt to nui th tch 2m3 Loi to S mu Mt (x 106 tb/ml) Mi cy T. chui C. vulgaris 4 3 0.320.04 0.90.18 Thu hoch 0.740.05 2.60.50 2 2 131.3 188.9 Thi gian nui (ngy) % tng trng

+ Thi gian nui 2 ngy + Tc tng trng T. chui t 131.3 % sau 2 ngy nui v t 188.9 % to C. vulgaris.

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+ To T. chui thu hoch th tch 2 lt mt phn dng lm ging nhn mi b ximng con ch yu l thu hoch trc tip cho lun trng n. + To C. vulgaris kt thc nui sinh khi th tch 2 m3, sau 2 ngy nui c thu trc tip vo cc b nui lun trng. 4..2.2.4 Nhn sinh khi to b xi mng Bng 16 Mt to nui b ximng Loi to S mu Mt (x 106 tb/ml) Mi cy T. chui I. galbana 2 1 0.180.01 1.750 Thu hoch 0.470.02 5.50 Thi gian nui (ngy) 3 3 % tng trng 161.1 214.3

+ B ximng ch hn ch nui 2 loi T. chui v I. galbana do iu kin kh qun l, v tng trng cng chm hn cc th tch nh ch t 161.1 % sau 3 ngy nui (to T. chui) trong khi th tch 1m3 to t mc tng trng 250.0 % sau 3 ngy nui. + Ging nui ch yu cc th tch ngoi tri l T. chui. + Qua kt qu theo di nhn thy tt c cc th tch nui ngoi tri to T. chui pht trin nhanh nht (khc vi trong phng th nghim). 4.2.3 Mt s ch tiu theo di

B tr th nghim theo di tc pht trin ca cc loi to nui v theo di bin ng mt s yu t mi trng. 4.2.3.1Theo di s tng trng ca mt s loi to ( th tch 2 lit) a, Tng trng Nanochloropsis oculata

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Mao t (x106 t/m l) 1 00 80


67.3 75.6 76.4

60 40
26.4 46.3 47.90

Mao t

20 0 0

10.5

15.6

Thgian i (nga) y 2 3 4 5 6 7

o 2 Tag trg N. oculata th n n

+ To N. oculata vi mt cy ban u l 10.50.5 (x 106 tb/ml) sau 6 ngy nui t mt cc i 76.43.77 (x 106 tb/ml) tng 627.6 % so vi ging ban u, t ngy th 7 mt to bt u gim. b, Tng trng Tetraselmis chui
Mao t (x 106 tbm l) 12 10 8 6 4 2 0 1 2 3 4 5 6 7 o 3 Tag trg T. chui th n n
2.4 3.9 6.0 8.2 10.5 9.3 7.9

Mao t

Thgian i (nga) y

+ To T. chui vi mt cy ban u l 2.4 0.18 (x 106 tb/ml) sau 4 ngy nui t mt cc i 10.51.33 (x 106 tb/ml) tng 337.5 % so vi ging ban u, t ngy th 5 mt to bt u gim nh, t ngy th 6 tr i l bt u pha tn. c, Tng trng Isochrysis galbana

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Mao t (x 106 tb/ml) 50 40 30 20 10 0 0 1 2 3 4 5 6 o 4 Tag trg I. galbana th n n


11.8 8.2 17.1 31.6 25.0 39.8 32.2

Mao t

Thgian i (nga) y

+ To I. galbana vi mt cy ban u l 8.2 0.17 (x 106 tb/ml) sau 5 ngy nui t mt cc i 39.83.68 (x 106 tb/ml) tng 385.4 % so vi ging ban u, t ngy th 5 mt to bt u gim v bt u pha tn. d, Tng trng Chlorella vulgaris
Mao t (x106 tb/ml) 60 50 40 30 20 10 0 0 1 2 3 4
6.6 10.0 17.5 32.0 51.8 48.8

Mao t

Thgian (nga) i y 5

o 5 Tag trg C. vulgaris th n n

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+ To C. vulgaris vi mt cy ban u l 6.6 0.19 (x 106 tb/ml) sau 4 ngy nui t mt cc i 51.83.89 (x 106 tb/ml) tng 684.8 % so vi ging ban u, t ngy th 5 mt to bt u gim v bt u pha tn. 4.2.3.2Theo di s bin ng pH trong cc b nui to (Vnui = 2 lt). Bng 17 bin thin pH cc loi to nui (trong phng th nghim) Loi to N. oculata T. chui I. galbana C, vulgaris
pH 9.6 9.4 9.2 9.1 9 8.9 8.8 8.6 8.4 Pha cha m tag trg n n dg n ca pha tag trg cu ta c n n a o 8.8 8.9 8.8 9.4 9.3 9.2 N. oculata T. chui I. galbana C. vulgaris

Pha bt u (1) 8.80 8.80.0 8.80.1 8.80.2

pha tng trng (2) 9.30.1 8.90.0 9.10.2 9.40.2

Pha dng (3) 9.20.1 8.90.1 8.80.2 9.20.1

o 6 Bie thie pH c loata kha nhau th n n ca i o c

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Mc pH thun li cho a s cc loi to l 7 9 v mc thun li nht l 8.2 8.7, nu pH qu cao hoc qu thp u tr thnh yu t km hm s pht trin ca to (Ukeles R, 1971). + Qua bng v th theo di bin thin pH trong cc bnh nui th tch 2 lt cho ta thy: - Vi mt ging ban u khc nhau N. oculata l 10.50.5 (x 106 tb/ml), T. chui l 2.4 0.18 (x 106 tb/ml), I. galbana l 8.2 0.17 (x 106 tb/ml) v C. vulgaris l 6.6 0.19 (x 106 tb/ml) nhng mc pH thi im ban u l tng i ging nhau tt c cc loi c gi tr l 8.8 0.2. - Khi cc loi to t gi tr cc i th gi tr pH bin thin rt khc nhau cc loi: N. oculata l 9.30.1, T. chui l 8.9 0.0, I. galbana l 9.1 0.2 v C. vulgaris l 9.4 0.2. - Khi to pha dng pH ca cc loi c xu hng gim nh, Ring T. chui hu nh khng gim (pH = 8.90.1). Nh vy - Mt to ban u khng nh hng ti pH trong cc bnh nui. - Trong cc bnh nui to pH bin thin t l thun vi mt to. - Ti thi im to t mt cc i, pH tng i cao so vi khong 7 9. 4.2.3.3Theo di s bin ng Ammonia trong cc b nui to Bng 18 Bin thin NH3-N cc loi to nui (trong phng th nghim) Loi to N. oculata T. chui I. galbana C, vulgaris Pha bt u (1) 0.0670.008 0.3870.169 0.1160.05 0.0210.011 pha tng trng (2) 0.0410.015 0.1030.049 0.7410.778 0.0130.016 Pha dng (3) 0.0560.013 0.0650.033 0.2950.111 0.0130.015

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NH3 -N g/l) 0 (m .8 0 .7 0 .6 0 .5 0 .4 0 .3 0 .2 0 .1 0

0 41 .7

0 8 .3 7 0.2 5 9 0 1 .1 6 0 6 .0 7 0 03 .1 0 41 .0 tan g trg n 0.0 6 0.0 5 5 d n g

N. cu ta la T. ch i u I. g lb na a a C. vu a lg ris

Ph ch a am

Cac p a h tan trg g n

o 7 Bie th n NH3 loai tao th n ie -N cac

4.2.3.4Theo di s bin ng pH v NH3_N ca to Tetraselmis chui cc th tch nui ngoi tri Bng 18 Bin thin NH3-N cc loi to nui (ngoi tri) NH3 Vnui (m ) 0.5 1 2 9.050 8.400 8.950
3

pH Ngy cy Thu hoch 9.350 8.700 9.250

_N Ngy cy 0.136 0.019 0.032 Thu hoch 0.141 0.134 0.074

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pH 9.6 9.3 9.0 8.7 8.4 8.1 7.8 Mca i y thu hoa h c
9.050 9.350

the ch 0.5 lt tio the tch 1 m3 the tch 2m3 thie o i m

o 8 Bie thie pH cu ta Tetraselmis chui c the th n n a o ca tch ngoath i i

+ PH cc b nui to ngoi tri cng bin thin t l thun vi mt to nui. + PH ca to T. chui cc th tch nui ngoi tri gi tr ln hn cc bnh nui 2 lt trong nh v cng c bin dao ng ln hn.

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H3_N N (mg/l) 0.2 0.1 0.1 0.1 0.1 0.1 0.0 0.0 0.0
0.032 0.019 0.074 0.136 0.141 0.134

the 0.5m3 tch the 1 m3 tch the 3 m3 tch Thie o i m

1 2 o 9 Bie thie NH 3-N cu ta Tetraselmis chui th n n a o ca the ngoatr c tch i i

4.2.3.5Theo di s bin ng nhit v mn ca cc th tch nui ngoi tri a, Theo di s bin ng nhit ca cc th tch nui ngoi tri
Nhieo t (0C) 38 36 34 32 30 28 26 24 22 20 06/6 10/6 14/6 18/6 22/6 26/6 30/6 04/7 o 9 Bie thie nhieo the 0.1 m3 th n n t tch

Sag n Chie u

Nga, thag y n

34

Theo Guillard R.R.I (1997) khong nhit thch hp cho s pht trin cc loi to l 20 24 0C v khong nhit m cc loi to c kh nng chu ng l 16 27 0C. + S nhn xt v bin thin nhit theo t, theo tng th tch. b, Theo di s bin ng mn ca cc th tch nui ngoi tri
o n () ma 36 35 34 33 32 31 30 29 15/6 17/6 19/6 21/6 23/6 25/6 27/6 29/6 01/7 Nga thag y n

0.1 m3 0.5 m3 1 m3 2 m3

o 11 Bie thie o n th n n ma

PHN V KT LUN V XUT

35

5.1

Kt Lun

+ iu kin quan trong nht trong lu gi ging l k thut v trng + To gingc th lu gi trong t gi to 6 8 tun nhng cht lng ging tt v to pht trin nhanh khi nhn ging th nn nui san tun th 3 4. + Trong cc loi to theo di th to Chlorella sp pht trin nhanh nht, tip l Nnochloropsis oculata, Isochrysis galbana cui cng l Tetraselmis chui. + Trong iu kin nui phng th nghim th tch nh to pht trin nhanh t mt cc i hn th tch ln. + Mt ging ban u cao to nhanh t mt hn. + Nui ngoi tri th tch nh to pht trin chm hn th tch ln. + Nhit trong ngy dao ng ln th nh hng ti s pht trin ca to. + mn qu thp hoc qu cao cng nh hng ti s pht trin ca to. + Bin ng ca pH t l thun vi tng trng ca to, khi to pht trin pH tng khi to tn pH gim. + Ammonia bin ng t l thun vi pH 5.2 Xut

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