I. T VN - Nng nghip ng vai tr rt quan trng trong nn kinh t nc ta.

. Trong sn xut nng nghip, ngi nng dn gp rt nhiu kh khn, trong kh khn ln nht l phng tr v tiu dit cc loi su hi cy trng. Theo thng k ca t chc lng nng th gii (FAO) cho thy: cc loi cy trng trn ng rung hin nay phi chng vi hn 100.000 loi su hi khc nhau, 10.000 loi nm, 200 loi vi khun, 600 loi tuyn trng v 600 loi virus gy bnh. Hng nm c khong 20% sn lng lng thc thc phm trn th gii b mt trng (Trn th Thanh, 2003). khc phc tnh trng trn, con ngi tch cc tm kim cc bin php phng chng cc tc nhn gy hi, nhiu bin php khc nhau c s dng, trong c bin php s dng thuc tr su ha hc, chng ng mt vai tr quan trng trong h thng canh tc nng nghip trong mt thi gian di v mang li hiu qu cao phm vi s dng rng ln. C th ni, khng mt bin php bo v ma mng no hiu qu hn bin php ha hc v mt qui m v hiu qu. - Nhng cc bin php ha hc bc l ngy cng nhiu nhng khuyt im ca n, sau khi dng cht dit c hoc thuc tr su ha hc, mi trng b nhim, con ngi b ng c v c khu h sinh vt i km cng b nh hng lm mt cn bng sinh thi. iu nghim trng hn l tnh trng gia tng liu lng v thi gian phun thuc ha hc chng su bnh to nn d lng thuc khng cho php trn rau mu v lng thc, l nguyn nhn gy nhim c cho khong 1,5 triu ngi mi nm trn ton th gii, trong c khong 25 nghn ngi b t vong (WHO, 1998). - Trc thc trng ny, cc nh khoa hc nng nghip nghin cu v a ra phng php mi trong phng tr v tiu dit cc loi su hi cy trng. Mt trong nhng phng php l kim sot sinh hc: nghin cu s dng nm, vi khun, virus, k sinh thin ch, cc hp cht th cp c ngun gc tho mcc kh nng phng tr, tiu dit su hi cy trng hiu qu v an ton. - Do , cc ch phm phng tr su hi cy trng c ngun gc sinh hc rt c quan tm, bi chng t nh hng n sinh vt sng, d phn hy, khng lm c nng phm v tin s dng. Trong , c cc ch phm t cy neem (cy xoan chu hn). - Cy xoan chu hn (Azadirachta Indica A.Juss) l mt trong nhng loi tho mc c c tnh khng su bnh ang c nghin cu v s dng ngy cng nhiu nc ta v mt s nc trn th gii do chng c kh nng phng tr hn 400 loi dch hi. Cy xoan chu hn c ngun gc t n , du nhp vo nc ta cch y gn 30 nm v c trng nhiu cc tnh min trung. Hin nay rng neem ti Ninh Thun v Bnh Thun vn khng ngng pht trin vi din tch hn 1.000 ha, y l ngun nguyn liu quan trng, bc u to ra cc sn phm phng tr su hi cy trng. 5

II. TNG QUAN TI LIU 2.1 Gii thiu chung v cy xoan chu hn 2.1.1 Phn loi - Nghnh : Angiospermatophyta (nghnh thc vt ht kn) - Lp - Nhm - B - H : Dicotyledoneae (lp hai l nm) : Dialypetalae (nhm cnh phn) : Rutales (b cam) : Meliaceae (h xoan) - Lp ph : Archichlamydeae (lp ph nguyn hoa b)

- B ph : Rutinae - H ph : Melioideae - Tc (chi): Melieae - Ging - Loi : Azadirachta : indica A. Juss

1.1.2 Ten chung Co rat nhieu ten chung cho cay neem, ac biet nhng nc co trong cay neem lau i. Bang 1.1 Mot so ten thng dung cua cay neem ST T 1 CHAU A, CHAU UC: An o (Limba, Limbo, Vembu, khoang gan 100 ten); Pakistan (Nimmi); Myanmar (Tamarkha); Srilanka (Kohomba); Thailand (Dadao India, Kwinin, Dao); Uc, Fijii, New Guinea (neem); Viet Nam (Xoan An o, 2 3 4 CHAU PHI: CHAU MY: CHAU AU: xoan chu han, neem) Nigeria (Babo Yaho); Tanzania (Warobaini) My (neem); Chau My La Tinh (Nim). c (Neim, Nim, Indischer Zedrach); Phap (Lilas de Indes, Margousier); Tay Ban Nha (Nim, Margosa); Anh (India Lilac, Neem).
6

Chau

Ten cay neem theo tieng a phng

1.1.3 ac tnh thc vat Neem (Azadirachta indica) la cay phat trien nhanh, thng co chieu cao ti 20 m, tham ch co the len ti 40 m ieu kien thuan li. Neem la cay co la xanh quanh nam, nhng ieu kien khac nghiet trong thi k kho han keo dai, cay co the b rung het la, canh b kho gay. He thong re gom co re cai phat trien manh va nhieu re phu. Re phu co the dai ti 18 m (Benge, 1989). Canh la moc so le, dai t 20 40 cm, phien la co mau xanh lat en am. Mot canh co en 31 la, moi phien la dai t 3 en 8 cm, cuong la thng ngan. La non co mau o en o lat. La kho cha 12,40 en 18,27 % protein tho, 11,40 23,08% x tho, 43,32 66,60% N-free extract, 2,27% - 6,24% ether extract, 7,73 18,37% khoang tong so, 0,89 3,96% can xi va 0,10 0,30 % phosphor [92]. Hoa mau trang, co mui thm, nam nach la. Chum hoa ru xuong va dai ti 25cm. Cum hoa phan nhanh 3 lp va co ti 150 hoa, ac biet co khi co ti 250 hoa. Hoa n dai 5 6 mm, rong 8 11 mm, nam canh dai 5 mm, rong 2 mm, hoa lng tnh, nh c chn trc, hoa c moc cung cho giong nh cac loai hoa tap tnh [32, 43, 106]. Qua nhan hnh tron hoac ovan. Trai neem khi chn co kch thc 1,4 - 2,8 x 1,0- 1,5 cm. Qua mau xanh khi con non, tr nen mau vang xanh hoac vang khi chn gia. Vo qua mong, v ang, mau vang nhat va co nhieu x. Phan tht qua gia day 0,3 0,5cm. Nhan hat co vo lua mau nau, hat thng co mot nhan, oi khi co en 2 hoac 3 nhan. Hat co kch thc 0,9 2,2 x 0,5 0,8 cm, nhan co kch thc 0,8 1,6 x 0,4 0,5cm. So nhiem sac the la: 2 n = 28; n = 14 [100].

(a) Qua va cay neem (b) Hoa neem Hnh 1.1 Cay, qua neem (a) va hoa neem (b) trong Ninh Thuan

(a) Qua neem chn Hnh 1.2 Qua neem

(b) Qua neem xanh bo doc

2.1.3 Ngun gc v s phn b ca cy xoan chu hn Xoan chu hn c xem l c ngun gc vng Assam v Burma. Tuy nhin, ngun gc chnh xc ca n vn cha c bit, mt s ngi cho rng xoan chu hn sng t nhin vng tiu lc a n ; nhng ngi khc li cho rng n thuc vngkh hn trn ton khu vc Nam , ng Nam bao gm Sri Lanka, Thi Lan, Malaysia v Indonesia Xoan chu hn c di thc vo Chu Phi t nhng nm u ca th k 20. Ngy nay, n c trng rng ri t nht 30 quc gia, c bit vng dc theo khu vc vnh ai pha nam sa mc Sahara. Trong th k 20, xoan chu hn cng c di thc n Fiji, Mauritius v nhiu quc gia thuc Trung, Nam M Xoan chu hn c di thc vo Vit Nam nm 1981 do GS. Lm Cng nh, mt nh lm hc Vit Nam. Nhn dp tham d hi tho quc t lm nghip v Vai tr ca rng trong s pht trin ca cng ng nng thn ti Senegal, Chu Phi, ng em ht ging xoan chu hn v trng ti vng t Phan Thit, sau cho nhn rng ra v trng ti Ninh Thun, Bnh Thun. ng cng l ngi t tn cho loi cy ny l cy xoan chu hn(Azadirachta indica A.Juss) phn bit vi xoan a phng (Melia azedarach) c trng ph bin nc ta 2.1.4 iu kin thch nghi v tng trng

Xoan chu hn l loi thc vt c ph thch nghi rng v chu c iu kin khc nghit tt hn nhiu loi cy khc. Cy vn c th pht trin mnh trn nhng vng t kh hn, ngho cht dinh dng, t c mn trung bnh hoc kim nng. Cy cng c th sng st trn t c pH thp, do trong l xoan chu hn c hm lng cht khong kh cao, c tc dng gim acid trong t. Cy xoan chu hn thch nghi tt nhng vng nhit i kh nng vi nhit c th cao hn 450C v lng ma trung bnh khong 400 1500 mm/ nm. Mc tng trng ca cy cng ty thuc vo cht lng t, cy thng pht trin nhanh trong 5 nm u, c th t chiu cao 4 m sau 5 nm v 10 m sau 25 nm trng.

+ Anh hng cua vung sinh thai len s tch luy azadirachtin cay neem Ket qua phan tch 256 mau c thu t 26 quoc gia tren the gii cho thay ham lng azadirachtin trung bnh trong nhan hat neem cac mau la 3,6mg/g va ham lng dau la 46,7%. Co s bien ong rat ln ve ham lng azadirachtin trong nhan hat neem gia cac vung a ly khac nhau, ke ca ngay trong mot nc va ngay trong mot vung, tham ch gia cac cay trong cung mot vung (Ermel va cs, 1986; Benge, 1989). S khac nhau nay co the do cac yeu to kh hau va tho nhng, cung co the do ieu kien thu hai, che bien va bao quan hat neem. Mot so tac gia cho rang hai yeu to nhiet o va o am co anh hng ln nhat en ham lng azadirachtin trong hat neem. Theo mot so tac gia khac th, cac yeu to kh hau, tho nhng lai khong gay anh hng trc tiep en ham lng azadirachti. Singh (1986) thong bao ham lng hoat chat gay ngan an trong hat neem vung at kho can cao hn vung ven bien co nhieu ma. Senegan, hat neem c thu hoach hai lan trong nam, t mot(vu chnh), vao thang 6-7, t hai vao thang 10-11. ieu kien kh hau kho trong thang 10-11 cho ham lng azadirachtin trong nhan hat cao hn so vi vu chnh, thang 6-7 (mua ma).
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Nghien cu s bien ong ham lng azadirachtin gia cac vung khac nhau cua An o cho thay, ham lng azadirachtin A bien ong t 556,9-3030,8 mg.kg-1 nhan hat neem. Ham lng azadirachtin B bien ong t 43,1-590,6 mg.kg-1. a phan cac vung at kho can, kiem va chua man co ham lng dau thap (< 35%). Cac ket qua nghien cu con cho thay co s bien ong ln ve ham lng azadirachtin gia cac ca the cay trong cung mot vung at. ieu nay chng to cac yeu to nh lng nc ma, am o va nhiet o khong gay anh hng nhieu en ham lng hoat chat azadirachtin trong nhan hat neem [47]. Nhn chung, van cha ro yeu to nao, ben ngoai hay ben trong anh hng en ham lng hoat chat azadirachtin, salannin va nimbin trong nhan hat neem. 1.1.6 Mot so benh thng gap tren cay neem 1.1.6.1 Benh do vi khuan + Pseudomonas azadirachtae (Pseudomonadaceae) la vi khuan gay benh cay neem, lan au c phat hien Bang Gujarat An o. No chnh la nguyen nhan lam nho la, mong nc, nhieu not goc la, lam vung xung quanh not mat la tr nen vang, chet tng vung va cuoi cung la kho heo. La b nhiem benh se m uc i va cac not phat trien thanh nhng van gia nh la b ve bua. Nhng la b nhiem khuan nang tr nen vang, heo kho va rung. + Xanthonas azadirachtti (Pseudomonadaceae) la nguyen nhan gay benh om nho, trang uc, mong nc, sau chuyen mau thanh nau nhat va nau am tren la. Nhng vet nay cang ngay cang ln va chet hoai t trung tam. 1.1.6.2 Benh do nam + Pseodocercospora subsessilis (=Cercospora subsessilis): La nam gay hai manh nhat cay neem, no la nguyen
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nhan gay benh om la vi trieu chng: mo b nhiem benh au tien co mau xanh sang va sau tr thanh mau nau. Cac la b benh thng b lo cho vi kch thc lo to nho khac nhau. Ngoai An o, benh nay con thay xuat hien Philippines, Myanmar, vung ong va Tay Phi (Somali, Benin), vung bien Caribe (cong hoa Dominica) va Nam My (Equador). Thi ky u va phat trien benh keo dai t 25 en 50 ngay. + Aspergilus spp.: Mot vai loai nam Aspergillus, ac biet la A. flavus, thng xuat hien hat neem khi bao quan. Nam ton tai dang bao t va phat tan rat nhanh, pha huy nhan va cac hoat chat sinh hoc trong nhan nh azadirachtin. A. flavus la nam moc nguy hiem nhat oi vi nhan hat neem v chung san xuat ra loai oc to mycotoxin va aflatoxin. Cac nam Aspergillus spp. khac co mat tren hat va nhan hat neem la A. niger, A. tamari, A. versicolor va A. quercinus (= A. melleus) (Sinniah va cs, 1983). 1.1.6.3 Benh ky sinh thc vat Dendrophtoe falcate, Tapinanthus sp. va Loranthus sp. c ghi nhan la nhng k sinh tren cay neem An o va chau Phi. 1.1.6.4 Benh roi loan chc nang Khoang vao nam 1990, xuat hien nhng bao cao ve benh roi loan chc nang cay neem trong Nam va Tay Nam Nigeria, Bac Nigeria va Mali. Cay neem nhng la tuoi khac nhau ong loat b rung la, cac mam cay va canh b rung lam nhieu cay b chet, tham ch ca nhng cay a c tren 20 nam tuoi. Ngi ta cho rang benh roi loan chc nang co le la do yeu to khong bnh thng cua moi trng, trong o s thieu hut nc hnh nh la nhan to chu yeu. Benh roi loan ve hap thu nc cua cay neem cac la tuoi khac nhau c phat hien mien Nam Yemen, vo cua cac cay nay b pha huy tng
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phan do b tan heo hay cac mam non cay b hong, la vang ua va rung, dch r tran ra than cay lam cay chet 1.1.7 Sinh trng va nang suat qua 1.1.7.1 Sinh trng Cay neem phat trien rat nhanh ieu kien thuan li. Cac yeu to nh pH at, kha nang gi nc cua at, lng nc ma, che o ti nc, chat dinh dng trong at, benh nam, yeu to di truyen va s canh tranh trong loai eu anh hng oi vi cay neem. vung at kho co o cao ln so vi mat nc bien va kh hau lanh, cay neem phat trien kem hn so vi cac vung khac. Theo Radwanski (1981), Nigeria, 66% s phat trien cua cay neem xay ra 3 nam au tien, cay co the cao 7 m, nam nam tiep theo, cay neem co the cao ti 11m. chau Phi, nhng vn co at tot, cay neem co the cao ti 1,5 m ngay nam au tien, nam th hai cao 2m, nam th 4 cao ti 4,5m vi ng knh than 7 8cm. Cameroon, cay neem 30 nam tuoi co ng knh than cay ti 1m. Cay neem nhieu nam tuoi vung at tot co the cao ti 30 35m vi ng knh than cay 2,5 3,5m. Nhng cay neem non can c bao ve khoi co dai moc lan, ac biet vi nhng cay neem mi trong. An o, tren thc te cay neem b co dai moc lan en 2 nam tuoi, khi cay a at chieu cao 0,5 en 1,4 m. Neu khong tr co dai thng xuyen, cay neem ch cao c 0,5 1,0 m. 1.1.7.2 Nang suat qua Cay neem khoang hai nam tuoi, a cho mot so qua au tien. Cay neem c ti nc ay u se phat trien nhanh va cho qua sm hn. Bac ban cau, thi k ra hoa va ket qua thng ri vao khoang thang 2 - 3 en thang 5 ( An o), va vung Nam ban cau vao khoang thang

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11 12. nhng vung trong nhieu cay neem, oi khi co nhng cay neem ra hoa va ket qua trai vu. mot so ni, hai mua cay neem ra hoa ket qua thng phan biet rat ro ret, mua au vao khoang thang 3 4, mua sau t thang 7 en thang 8. Thong thng cay neem ra qua vao khoang thang 7. mien Trung Senegal, s tro hoa cua cay cung co s khac biet gia cac cay trong cung mot vung, chu yeu do anh hng cua lng nc ma, ac biet trong tuan au tien cua mua ma. Nang suat qua cua cay neem thng bien ong, theo Koul O. (1990) mot cay neem cao 8 m, cho trung bnh t 35 55kg qua/ nam. Con pha Bac Nigeria, trung bnh mot cay cho khoang 20,5 kg. Nicaragua, nang suat qua cua cay 69 nam tuoi la 19,2 kg. Cay neem trong biet lap hoac trong tng nhom nho thng co nang suat cao hn cay trong tap trung. Saxena (1993) thong bao cay neem trng thanh cho trung bnh 25 kg qua/nam va co the thu en 100 kg qua nhng cay moc rieng le vung bien Kenia. Nhng th nghiem cua hang Baritiya cho thay cay neem t 8 10 nam tuoi cho nang suat qua 9 kg/nam, cay 15 nam cho 13 kg/ nam, cay 20 nam cho 19 kg/ nam. Nicaragua ch khoang 4% hoa au trai. Ty le trong lng gia hat va tht qua la 1:2; 1.800 hat nang trung bnh 1kg, nhng co s bien ong ln do hat co nhieu c khac nhau, tuy cay.
2.1.5 Nhn ging Xoan chu hn d c nhn ging bng c sinh sn hu tnh ln v tnh. Cy c th c trng t ht, t cy ging con, cy non, t chi r mt hoc nui cy m. Tuy nhin, hin nay cy thng c trng t ht. Do t l ny mm ca ht rt bin ng (15% i vi ht d tr v 85% i vi ht ti), nn nhiu chuyn gia khuyn co gieo ht sm trong vn m. Ht cn ngm trong nc lnh 24 gi v ct u v ht tng kh nng ny mm. Gieo ht trn nhng lung ct mn su 2,5 cm v cch nhau 2 5 cm. Ht thng ny mm t 1 2 tun.

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Nhiu tc gi cho rng ht khng tn ti c lu, ch sau 2 6 thng bo qun ht s khng th ny nm c. Tuy nhin, mt s nghin cu gn y Php cho thy, ht c bo qun m khng b h v qu trong (ni qu b) c kh nng ny mm n 45% sau 5 nm bo qun.

1.1.8.2 Chon giong cay cho nang suat qua va hoat chat cao Chon, nhan va lai giong e tao cay neem co nang suat qua vi ham lng dau va hoat chat azadirachtin cao van cha at ket qua kha quan do ham lng azadirachtin luon bien oi (Ermer va cs, 1983). Ham lng azadirachtin bien ong phu thuoc vao ieu kien moi trng hn la cac yeu to di truyen. S dung k thuat di truyen va nuoi cay mo e phat hien va nhan giong neem co kha nang khang sau benh hay co ham lng azadirachtin cao cung mi ch c bat au (Rice, 1993).
2.3 Cc hot cht phng tr cn trng trch t cy xoan chu hn 2.3.1 Cc hot cht c trong xoan chu hn Trong cy xoan chu hn, trn 300 hp cht t nhin c phn lp v m t. Tanin c nhiu trong v cy (14%), l cy ngoi cc limonoid cn cha tinh du v alkaloid. Theo Mitra (1963), l cy cha alkaloid Parisian, xoan chu hn t bo v n khi nhiu loi cn trng gy hi do n cha nhiu thnh phn c hot tnh khng cn trng. Xoan chu hn cha hn hp ca 3 n 4 hp cht chnh v khong hn 20 cc hp cht ph khc, cc hp cht ph ny c tc ng theo phng cch ny hay phng cch khc. Cc hp cht chnh thuc v nhm cc hp cht t nhin triterpenoid; chuyn bit hn, l cc limonoid (tetranotriterpenoid) Cho n nay, 9 limonoid trong xoan chu hn c chng minh l c kh nng ngn cn s pht trin ca cn trng, c bit l nhng cn trng gy dch bnh cho nng nghip v nh hng xu n sc khe con ngi. Nhiu limonoid mi vn ang tip tc c pht hin trong cy, cc limonoid nh azadirachtin, salannin, meliantriol, nimbin, nimbidin c xem l thnh phn hot cht chnh trong xoan chu hn. 2.3.2 Du xoan chu hn Theo Tewari (1992), nhn ht xoan chu hn c cha khong 40,0 48,9% du, du xoan chu hn c mu vng nu, mi ti. Theo Ketkar (1976), nhn ht xoan chu hn khi c chit xut bng phng php p thu c t 30 40% du. Cn du c th c chit xut bng mt s dung mi khc. Theo Anon (1985), du xoan chu hn c th c tch chit bng cch un si nhn ht xoan chu hn c xay nh trong nc. 14

2.3.3 Azadirachtin v cc limonoid khc 2.3.3.1 Azadirachtin Azadirachtin l hp cht chnh trong xoan chu hn, c cng thc phn t l C35H44O16 , nhit nng chy 154 1580C, c hot tnh khng cn trng mnh nht, c bit l tc ng xua ui v c ch sinh trng mnh. Azadirachtin c cu trc tng t nh hormone ecdysone, hormone ny c tc dng kim sot tin trnh bin i ni ha hc ca cn trng khi cn trng chuyn t dng u trng sang dng nhng sang dng trng thnh. Azadirachtin c xem nh cht ngn cn s tng hp cc hormone cn thit cho c th cn trng, do ph v chu k sng ca cn trng. Azadirachtin tp trung nhiu nht trong nhn ht xoan chu hn. Trung bnh 1 gram nhn ht cha t 2 n 4 mg azadirachtin, c bit Senegal, 1g nhn ht c th cha n 9 mg azadirachtin. Hm lng azadirachtin trong nhn ht xoan chu hn cng bin ng ty thuc vo ngun gc, xut x, iu kin kh hu nhiu vng khc nhau, trong mi cy cng c s bin ng v hm lng azadirachtin (Bng 2.2), (Ermel v cng s, 1987; Singh, 1987) Bng 2.2: S bin ng hm lng azadirachtin trong mu ht t nhiu ni khc nhau

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Hm lng azadirachtin cng ph thuc vo ti im thu hi, qu xoan chu hn thu hoch tt nht khi n chuyn sang mu vng hay vng xanh, lc ny hm lng azadirachtin cao nht. khng nn qu qu chn v rng v ht thng b gim cht lng khi b ri xung t.

Hnh 2.1: Cng thc cu to ca Azadirachtin 2.3.3.2 Meliantriol Meliantriol l mt hp cht thuc triterpenoid alcohol, l tc nhn gy ngn n mnh i vi cn trng ngay c khi x l vi nng rt thp. Nhiu loi cn trng khng dm n trong khong 2 6 tun sau khi tip xc vi ch phm t xoan chu hn

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Hnh 2.2: Cng thc cu to ca meliantriol 2.3.3.3 Salannin Salannin cng c hot tnh gy ngn n mnh v l cht chng li s lt xc ca cn trng. Mt s nghin cu ch ra rng cht ny tc ng mnh ln s lt xc ca con b cnh cng trn da chut Acalymma vitta v b cnh cng Nht Bn Popillia japonica. Trong ht, hm lng salannin thng t 15 1247 g/ g nhn ht xoan chu hn (Eeswara v cng s, 1996)

Hnh 2.3: Cng thc cu to ca Salannin 2.3.3.4 Nimbin v Nimbidin Hai cht ny c xem l c hot tnh khng virus mnh. Chng t ra c hiu qu i vi virus gy bnh trn cy c chua, bnh u ma v bnh trn gia cm. Eeswara v cng s (1996) bo co rng nimbidin c trong ht xoan chu hn t 9 n 619 g/ g nhn ht Nimbidin l thnh phn ch yu gy ra v ng ca dch chit t ht xoan chu hn vi cn. Nimbidin chim khong 2% trong nhn ht xoan chu hn 17

Hnh 2.4: (a) Cng thc cu to ca Nimbin (b) Cng thc cu to ca Nimbidin 2.3.3.5 Cc cht khc Nhng cht c bn cht limonoid mi c tm thy sau ny: Diacetylazadirachtin c chit t qu ti, hiu lc tng t azadirachtin. Hai cht tng t salannin l 3 diacetylsalannin v salannol gy ra s ngn n cn trng.

1.2.5 C che tac ong cua azadirachtin len con trung Azadirachtin anh hng len sinh trng, phat trien, bien thai, sinh san cua con trung. Nhng c quan khac nhau cua con trung c xac nh la ch tac ong cua azadirachtin theo c che thu quan hoa hoc. Cac tac ong nay co lien quan en qua trnh kiem soat noi tiet to than kinh cua qua trnh lot xac va thay long. Nhng c che tac ong phan t van cha ro 1.2.5.1 C che thu cam hoa hoc Viec chon cac cay chu cua con trung an tap phu thuoc vao cac chat c tao ra t cay chu, trong o co nhng chat c che va kch thch, chung tac ong phu
18

thuoc lan nhau. Thc an c con trung la chon phu thuoc vao he thong thu cam hoa hoc. Cac th nghiem a chng minh con trung co nhay cam ac biet oi vi azadirachtin so vi cac chat khac. Azadirachtin tac ong trc tiep len thu cam hoa hoc, thong qua he thong than kinh, gay c che tiep nhan thc an va tao cam giac gay ngan an con trung. Mot so th nghiem tren te bao con trung canh vay (Spodoptera frugiperda, Sf 9) nuoi cay mot lp cho thay azadirachtin c che mien phan chia te bao va s hnh thanh lp te bao, lam giam s tong hp protein. Cac kch thch c che do azadirachtin cung rat co y ngha oi vi nhng loai con trung an tap e trng. Dch chiet nhan hat neem va dau neem co tac ong km ham s e trng nhieu loai con trung. 1.2.5.2 C che noi tiet Nhieu nghien cu cho thay azadirachtin tac ong trc tiep len ni phong thch hoc mon ot ngc trc (PTTH) t the hach than kinh, ngan can phong thch hocmon lot xac t vong hach than kinh nao, lam giam vong tuan hoan va tac ong len tong hp hoc mon PTTH (S o 1.1). Azadirachtin con tac ong len qua trnh d hoa hoc mon lot xac con trung, hoc mon lot xac c chuyen thanh dang hoat ong hn la 20- hydroxyecdysone do ecdisone -20- monoxygenase xuc tac. V vay, qua trnh nay la kieu tac ong na cua azadirachtin va ong vai tro mot phan gay oc tong the oi vi con trung. Tren au trung Galleria mellonella, azadirachtin tac ong kch thch tao hoc mon tre hoa va lot xac, gay s lot xac qua mc do giai phong allatotropin thanh the allata, pha v qua trnh tao vo chitin va dan en thay oi vong sinh trng, lam con trung khong sinh san c va chet. au trung Lucilia cuprin, azadirachtin co tac ong gay thoai hoa nhan, tao khoi d nhiem sac to va s thoai hoa te bao, anh
19

hng trc tiep len s gian phan trong cac a phoi, te bao bieu b, te bao bieu mo ruot gia, gay chet te bao trong qua trnh phan chia giai oan sau trung ky.

S o1.1 Tac ong km ham trc tiep len he noi tiet con trung cua azadirachtin (BS. Parmar va C.M. Ketkar, 1996)
(*): iem xay ra tac ong km ham trc tiep cua azadirachtin len cac mo, enzyme va cac noi quan khac.( PTTH = hoc mon ot ngc trc, JH = hoc mon tre, TB mau = te bao mau, TH chitin = tong hp chitin)

1.2.5.3 Cac c che sinh ly khac Azadirachtin tac ong manh len he thong mo c cua con trung, lam giam kha nang van ong chung va kha nang bay thong qua c che bai tiet hoc mon tao m the hach than kinh. C ruot gia va ruot sau cua con trung b azadirachtin tac ong lam phong lean, thc an i
20

qua ruot rat cham, lam con trung roi loan tieu hoa, roi chet. Ngoai ra, azadirachtin con tac ong lam giam bai tiet cac men phan giai protein, gay benh hoai t cac te bao bieu mo ruot con trung. Tac ong cua azadirachtin len cac ap ng mien dch c thay R. prolixus, khi x ly vi lieu 1 g/ml azadirachtin lam giam ap ng mien dch trong 6 ngay. Co the do thong qua c che noi tiet, azadirachtin ieu hoa he thong mo te bao mau, lam giam nang lc tong hp protein con trung.

Hnh 1. Tc ng gy bin dng ca neem i vi ry nu a , b : Ry nu cnh di v cnh ngn bnh thng. c, d, e, f: Cc kiu ry bin dng

Hnh 2. Tc ng gy bin dng ca neem i vi ng i go a, b, e: u trng, nhng, thnh trng bnh thng. c, d: u trng, nhng d dng. f, g, h: Cc kiu bin dng ca th nh trng

21

Hnh 3. Mt s tc ng ca neem i vi su t a, b: Tc ng gy ngn n (a l i chng, b l mu x l neem) c, d: tc ng lm bin dng thnh trng; e: tc ng gy cht nhng

1.2.6 Moi tng quan gia hoat tnh va cau truc Cac nghien cu co he thong ve moi tng quan gia cau truc va hoat tnh, th nghiem th hoat tnh tren tuyen trung, vi khuan, nam, nhuyen the va cac loai khac hien cha nhieu. Cac th nghiem hoa dc chu yeu c th bang dch chiet tho . Mot so th nghiem khac cho thay, nhng chat khong co cau truc tng t nh salannin th hau nh khong gay ngan an. Goc ester gan vi C-4 c cho la ong vai tro quan trong oi vi hoat tnh gay ngan an cua azadirachtin, v tat ca cac dan xuat khong co goc ester th ch co hoat tnh c che sinh trng, hoat tnh gay ngan an.
3.2.2 Phng php chit xut th hot cht sinh hc t nhn ht xoan chu hn Nguyn liu Qu xoan chu hn c thu hi t cy 4 5 tui trng ti Ninh Thun, lm sch v v tht ht, sau phi kh ht trong bng rm hoc sy kh nhit 500C. Ht c tch v ly nhn ht p du. Thit b v ho cht My p du thc vt chuyn dng KOMET Model D 85 1G (c). My c quay chn khng. Dung mi tanol. Cch tin hnh 22

khong co

Nhn ht xoan chu hn c p lnh trn my p du chuyn dng Komet (c) nhit c khng ch t 350C n 450C, thu c du v bnh du. Bnh du (b cn li sau khi p du) c cho ngm kit v chit xut vi ethanol nhm tn thu cc hot cht cn li trong bnh du, c bit l azadirachtin. Dch chit bnh du c loi ht ethanol bng c quay chn khng nhit 500C v trn vi du, thu c du xoan chu hn lm giu azadirachtin. (Hnh 3.1)

Hnh 3.1: Qui trnh chit xut th hot cht sinh hc t nhn ht xoan chu hn

23

Hnh 3.2: My Micro Kjeldahl

Hnh 3.3: My c quay chn khng

Hnh 3.4: Lc chn khng

Hnh 3.5: H thng sc k lng cao p (HPLC)

Hnh 3.6: H thng Soxhlet

Hnh 3.7: My p du chuyn dng KOMET (c)

u im ca cc dch chit t xoan chu hn 24

So vi thuc tr su tng hp, u im ca thuc tr su gc tho mc l kh nng lm chm s pht trin tnh khng tc nhn gy dch hi. Trong s cc thuc gc tho mc ph bin, dch chit t cy xoan chu hn phong ph hn c v thnh phn hot cht. Pyrethrin (sn phm chit xut t cy hoa cc Chrysanthemum cinerariaefolium) ch cha 4 loi ester v Rotenone (sn phm chit xut t cy thuc l Derris spp. v Lonchocarpus spp.) ch cha khong 6 loi isoflavonoid c hot tnh dit cn trng. Cn cy xoan chu hn, ring nhm azadirachtin gm khong 9 loi ng dng khc nhau, trong nhm azadarachtin c cho l c hot tnh khng cn trng mng nht. Ngoi ra, cha k cc loi limonoid khc cng c hot tnh cao nh salanin, meliantriol c mt trong v v l xoan chu hn. Hot lc ca cc dch chit t xoan chu hn c to thnh t s phi hp tng tc phc tp ca nhiu thnh phn hot tnh, nh lm gim tnh khng ca cc loi dch hi v s nhy cm v tp tnh (Isman, 1997). Th nghim chn lc trong iu kin phng th nghim trn loi rp Myzus persicae nhn thy rng: khi x l azadirachtin tinh sch ln cy th tnh khng ca loi rp ny tng ln 9 ln sau 40 th h, nhng x l bng dch chit ht xoan chu hn (cha hm lng azadirachtin tng ng) th khng lm xut hin tnh khng ca M. persicae sau chng y th h. C th chnh s a dng v thnh phn hot cht ca dch chit ht xoan chu hn lm phn tn tin trnh chn lc thch nghi ca i tng dch hi, nh lm chm s pht trin tnh khng ca chng. Tuy nhin, cng nh bao nhiu thuc tho mc khc, cc sn phm chit xut t xoan chu hn d b phn hy v mt sinh hc, khng bn vng trong mi trng t nhin do d b oxy ha, khng lm hi thin ch v khng gy c hi i vi ngi

1.7 Mot so ng dung

Qua va la neem c s dung e tao nhieu san pham. Mot cay neem a phat trien ay u co the cho tren 50 kg qua va 350 kg la moi nam. T qua co the thu nhng san pham phu di ay

25

S o1.3 Thanh phan qua neem (B.S. Parmar va C.M. Ketkar, 1996) 1.7.1 ng dung lam thuoc tr sau hoa nong

S o 1.2 Vai tro tham gia cua cac san pham t cay neem trong quan ly dch hai tong hp (BS. Parmar va C.M. Ketkar, 1996).

26

Cac san pham c che bien t hat va la neem eu co hoat tnh tr sau, nam gay benh, tuyen trung, gay ngan an, c che phat trien, c che sinh san, giam kha nang e trng, lam bien thai nhieu loai con trung va co ac tnh xua uoi. Tren 300 loai con trung c kiem soat bi san pham t neem. a co mot so thuoc bao ve thc vat c san xuat va thng mai da tren nen chnh la dau neem va cac hoat chat chiet t nhan hat neem, la neem. Rieng An o a co ti 100 san pham t cay neem, tnh en nam 1996

1 7.2 Mot so phng phap san xuat thuoc tr . sau dang tho t hat neem
27

+ Dch chiet nc: Hat neem c phi kho, lam sach, tach vo lay nhan hat, nghien nho va hoa vi nc. e qua em, loc qua vai va phun trc tiep len rau (che pham nay khong tron them bat c mot phu gia nao khac), lieu dung la 800 lt/ha. + Dau neem: Dau neem thu c bang ep tay hay bang may ep chuyen dung KOMET(c), s dung vi lieu khuyen cao t 5 10 lt/ha. + Bot nhan hat neem: Bot nhan hat neem cung c chuan b nh dch chiet nc tren, vi lieu dung 3 5g/ lt nc. Cac san pham tho nay c dung e phong tr Scrobipalpa ergasima tren rau xanh a phng( Togo), phong sau hai jacobielia faciallis va Lepidopterous tren ca tm, tr Plutella xylostella va Helluta undalis tren bap cai. + Che pham dau neem c lam giau Azadirachtin ( AZTVRT-K) va dch chiet nhan hat neem: Nhan hat neem c nghien nho, ngam nc vi ty le 25g/ lt trong 24 gi, loc vai. Dch sau loc c dung phun trc tiep. Che pham AZTVRT-K c dung dang huyen phu (EC) c hoa vao nc vi nong o thch hp (lieu khuyen cao, 162 lt/ha). Che pham nay c dung e phong Heliothis spp. va Plutella xylostella bap cai, hieu qua tng ng vi thuoc ac tr hoa hoc Tamaron 600 . 1.7.3 Mot so ng dung trong y hoc Dch chiet la, hat va dau neem c s dung pho bien e phong tr mot so benh trong y hoc Bang 1.5 Mot so san pham thng mai dung trong y hoc San pham Sensal JK-22 Thanh chnh Dau Dch chiet la
28

phan Cong dung Cai e ai ng

Ni

san

xuat An o An o

Greneem Neemcur e Pasutone Ioquin

Dch chiet la Dch chiet dau neem Bot la, hat Dch hat chiet

Loc mau la, Chong nhiem trung Tay giun, san ngi la, Tr benh ret sot

An o An o An o An o

1.7.4 Mot so san pham khac Dau neem va dch chiet la neem c phoi tron vi mot so phu gia e san xuat mot so loai my pham nh kem dng da, kem chong muoi, xa bong ve sinh, thuoc anh rang, v.v. .

1.8 Tnh hnh nghien cu cay neem Vit Nam

Nam 1981, giao s Lam Cong nh a tien hanh trong th nghiem cay neem giong Senegal tnh Ninh Thuan. Bc au cho thay cay neem phat trien tot, thch nghi vi vung at kho can ay. Con cay neem giong An o c du nhap vao nc ta nhng nam sau nay [5, 6]. T au nhng nam 1990, viec trong neem phat trien manh 2 tnh Ninh Thuan, Bnh Thuan la ni co kh hau kho han nhat nc ta, vi lng ma trung bnh hang nam ch vao khoang 500 mm va ch tap trung vao 1-2 thang trong nam. Ninh Thuan, cay neem a c quy hoach trong thanh rng doc theo vung at cat ven bien. en nay, a co tren 5.000 ha cay neem, trong o co hn 700 ha a cho qua. Theo anh gia ban au, cay neem a thch nghi vi vung at kho can Ninh Thuan. Trc mat, viec trong cay neem phuc vu cho muc ch phu xanh at trong, cai tao moi trng song va ban au tao nguon nguyen lieu lam thuoc bao ve thc vat. cac tnh pha Bac, a co mot so cong trnh tach chiet hoat chat t hat neem e lam thuoc tr sau (Dng Anh Tuan, Vien Hoa hoc, 2003). Theo anh gia chung, cac tac gia eu cho thay: Thuoc tr sau thao moc t neem
29

tuy khong manh bang thuoc tr sau hoa hoc nhng pho tac ong rong, khong gay ln thuoc, thi gian tac ong keo dai, ac trng cho thuoc tr sau sinh hoc. Cac nghien cu nay con b han che do nguon nguyen lieu t v cay neem to ra khong thch hp vi cac vung at va kh hau cac tnh pha Bac (cay a c trong nhng phat trien cham, khong ra hoa, ket trai). cac tnh pha Nam, viec trong va phat trien cay neem co thuan li hn, nen san co nguon nguyen lieu la hat neem e nghien cu sau hn ve k thuat trong va tach chiet hoat chat t neem. Nam 2001, Vien Sinh hoc Nhiet i c Nha Nc giao thc hien e tai theo ngh nh th gia Viet Nam va An o ve Nghien cu va s dung cay xoan chu han (Azadirachta indica), tc cay neem, trong tai Viet Nam. Ket qua, Vien a hoan thien c quy trnh nhan giong cay neem bang nuoi cay mo va a tien hanh th nghiem trong cay neem tren dien rong huyen Ninh Phc, tnh Ninh Thuan. Vien Sinh hoc Nhiet i a xac nh c quy trnh chiet tach tho hoat chat sinh hoc t nhan hat neem, quy trnh tach, tinh sach azadirachtin, salannin va xac nh ham lng azadirachtin, salannin va nimbin bang k thuat sac ky long cao ap (HPLC), khao sat anh hng cua dch chiet tho nhan hat neem len mot so loai nam gay benh va ray nau hai lua. Mot so tac gia khac cua Vien Sinh hoc nhiet i nh Nguyen Th Quynh (2000) a th nghiem che pham dau neem Plasma cua An o phun len day khoai lang trc khi trong cho thay co tac dung lam giam ty le nhiem ha cu khi thu hoach. Nguyen Th Phng Thao (VSHN, 2000) cung s dung che pham tren e bao quan cu khoai lang, ket qua thu c cho thay dau neem co tac dung xua uoi bo ha k sinh lieu x ly 200 ppm [16, 17]. Nhng nam sau nay, mot so nha nghien cu a tien hanh th nghiem
30

san xuat thuoc BVTV t hat neem cho ket qua kha quan nh: Vo Van Kim (Hiep hoi rau qua a Lat, 2005) a san xuat c 3 dang che pham t neem nh Neemcide 3000 EC dang nhu dau, Neemcide SP dang bot rac va Neemcide 3000ES dang dung dch vi tong ham lng limonoid (AZRL) 3%, co tac dung diet moi, ve, bo chet, rep en, bo tr hai tra [12]. Ngo Van at va cs (2005) nghien cu trch ly hoat chat azadirachtin t hat neem e gia cong thanh che pham Vineem 1500 EC, che pham co tac dung tr bo tr, rep sap hai nho t 68,9% en 75,5% sau 7 ngay phun, tr sau xanh bm trang hai cay bap cai c 60% - 90% sau 5 7 ngay phun [4]. Tran Kim Quy (2005) a xay dng c quy trnh chiet xuat limonoid t la neem bang con 950 e san xuat thuoc BVTV Neemcide SP 3% va t hat neem, bang phng phap ep nguoi khung ban, thu c dau va cao chiet co ham lng AZRL 1,61% lam nguyen lieu san xuat thuoc Neemcide SC 3%. Cac che pham thuoc BVTV nay hien ang trong giai oan th nghiem .

2.7. Tao che pham phoi tron gia dau neem va Bt (Bacillus thuringiensis)
Che pham c chuan b dang ti e dung ngay. Dch chiet banh dau cha 1600 mg azairachtin/lt dch chiet, che pham Bt dang bot do Trung Quoc san xuat co mat o bao t 3,2 x109 bao t/mg. Phoi che thanh 16 cong thc (Bang 2.1) va th nghiem hieu lc cac cong thc tren sau xanh va sau t trong phong th nghiem. Bang 2.1 Cong thc phoi tron dau neem va bot Bt
TT Cong thc phoi che Thanh phan chnh (%, m/v) Dau Bot neem (% Bt (% 31 TT Cong thc phoi che Thanh phan chnh (%, m/v) Du Bt Bt neem (% (% m/v)

1 2 3 4 5 6 7 8

N0B0 N0B1 N0B2 N0B3 N1B0 N1B1 N1B2 N1B3

m/v) 0 0 0 0 16,4 16,4 16,4 16,4

m/v) 0 5 10 15 0 5 10 15

9 10 11 12 13 14 15 16

N2B0 N2B1 N2B2 N2B3 N3B0 N3B1 N3B2 N3B3

m/v) 32,8 32,8 32,8 32,8 49,2 49,2 49,2 49,2

0 5 10 15 0 5 10 15

2.8 Tao che pham Neemaza va Bimectin dang nhu dau

e tao ra san pham Neemaza va Bimectin 0,5EC co kha nang nhu hoa tot, can tien hanh khao xat ty le phoi tron gia ham lng chat tao nhu va cac chat phu gia e tm ra cac thong so toi u cho qua trnh san xuat. Thanh phan cac chat c s dung c trnh bay trong bang 2.2. Ty le phoi tron phu hp cua tng chat se c trnh bay trong phan ket qua va thao luan. Bang 2.2 Thanh phan va ty le cac chat s dung tao TBVTV t hat neem ST T 1 2 3 4 5 Thanh phan Chat tao nhu Dau neem Chat tac ong phoi hp Phu gia khac Dung moi them cho u Ham lng (%) v/v v/v w/w v/v 100%

2.9 Xac nh tnh chat hoa ly cua che pham

2.9.1 Xac nh t trong che pham (10TCN 499 2002) + Xac nh khoi lng nc cat: can bnh t trong sach, kho (ca nut) chnh xac en 0,001g (mo), o ay nc cat un soi e nguoi vao be on nhiet (nhung ngap en co bnh) nhiet o 20oC t nhat 20 phut, lay bnh t trong ra, ay nut, lau kho, can va xac dnh khoi lng m1. Khoi lng nc 20oC c tnh bang gam theo cong thc:
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mn= (m1 mo) Trong o: mo - khoi lng bnh t trong, g m 1 - khoi lng bnh t trong va nc cat, g. + Xac nh ty trong che pham: T trong cua che pham 20oC (d20) c tnh bang g/ml theo cong thc: d20
n d 20 m2 = mn

n Trong o: d 20 - t trong cua nc 20oC (=0,9982 g/ml)

m2 - khoi lng cua che pham 20oC (phng phap xac nh khoi lng nh vi nc cat). 2.9.2 Xac nh o nht (CIPAC, 1999) [39] Dung cu va hoa chat: Nht ke, ong ho bam gi, ong ong, nc cat. Tien hanh: Tien hanh xac nh thi gian chay cua nc va cua che pham.

= o o Trong o: o can o (Centipose), (giay) o : thi gian chay cua nc (giay) 2.9.3 Xac nh pH dung dch 5% cua che pham Bc 1: pha nc cng chuan theo TCVN 3714 82 Bc 2: ong 95ml nc cng, a c chuan b bc 1, vao coc thuy tinh, sau them 5ml che pham cho u 100ml. Dung ua thuy tinh khuay eu, dung may o pH e o pH cua dung dch. 2.9.4 Xac nh o t nhu (TCVN 3714-82, CIPAC, 1999) o 100ml nc cng vao ong ong dung tch 100ml. Dung pipette nho 3 giot thuoc sat mat nc. ay kn ong ong, lac manh len xuong 2-3 lan roi quan sat xem chat
33

: :

o nht cua nc nhiet o thi gian chay cua che pham

long co c chuyen thanh nhu eu an va co nhng hat ln hay khong. 2.9.5 Xac nh o ben nhu tng (TCVN 3714-82, CIPAC, 1999) o t 75ml en 80 ml nc cng vao coc thuy tinh. Dung pipette them 5ml che pham (va them va khuay khoang 4 vong/giay). Nhu dau c them vao nc trong khoang 12 giay. Khi them nc, au pipette phai cach ay coc khoang 20 cm sao cho mau c rot thang vao gia khoi nc cng. Them nc cng en vach 100 ml. Trong luc them nc phai khuay lien tuc. Sau o chuyen ngay nhu dau vao ong ong 100ml sach, kho. Ghi lai be day lp kem sau 30 phut va sau 1 gi, 2 gi, 24 gi. Dung dch nay c gi lai e th o tai nhu. 2.9.6 Xac nh o tai nhu (TCVN 3714 82; CIPAC, 1999) Sau khi th o ben nhu tng, e yen ong nghiem trong 24 gi. Sau o lat ngc ong nghiem, lac 10 lan, quan sat xem o tai nhu co ben khong. Tiep tuc e yen 30 phut, quan sat lp kem tren be mat hay ay ong tao thanh day khong qua 4 ml la c.

2.10 anh gia o on nh cua thuoc bang phng phap gia toc (theo TCVN 10TCN 499 2002; CIPAC, 1999)
ay la phng phap anh gia o on nh cua thuoc nhiet o cao, cach lam nh sau: cho 50 ml mau th vao lo, ay mieng lot polyetylen, at lo vao tu on nhiet 54oC. Sau 30 phut, ay chat nut xoay vao lo va e lo trong ieu kien nhiet o 54oC trong14 ngay. Sau o lay lo ra, m nut xoay, e lo nguoi t nhien nhiet o phong roi ay nut xoay lai. Xac nh cac ch tieu ly hoa cua mau th trong vong 24 gi ke t khi lay mau ra khoi tu on nhiet. Theo
34

tieu chuan CIPAC, o on nh san pham ieu kien nhiet o 54oC trong14 ngay tng ng vi 2 nam bao quan san pham ieu kien t nhien.
3. Phng php xc nh c tnh ca du neem thng qua ch s LC 50 trn su xanh. Nguyn tc: S dng gi tr LD50 hoc LC50 ca cc cht gy c trn cng mt i tng sinh vt so snh c tnh ca chng. Th c tnh: du neem c nh ha bng tween 80, t l tween/du l 1/10. Sau pha long bng nc ct to hn hp c nng du neem nh mong mun. xc nh gi tr LC50, hn hp du neem c pha long thnh dy c 5 nng khc nhau (tng ng vi 5 nghim thc), cc nng a ra u c thm d trc m bo t l cht ca u trng su xanh tui 2 sau 5 ngy nm trong khong 16-84% gip cho vic xc nh gi tr LC50 sau 5 ngy c tin cy cao X l s liu: Tnh t l cht tch ly ca su sau 5 ngy mi nghim thc. T tnh gi tr LC50 sau 5 ngy ca hn hp du neem bng phng php phn tch Probit, thao tc trn phn mm Excel

Hieu lc thuoc c tnh theo cong thc Abbott Hieu lc (%) = (1Trong o: Ta : so ca the sau o x ly thuoc sau khi x ly Ca : so sau chet o oi chng sau khi x ly.
Ta Ca Ta ).100 = ( ).100 Ca Ta

35