I TiNG IIAN & NGHIEN ctfu I
Kha nang diet Su trung mu§i ciia dich chiet md seo cay cue ao Spilanthes acmella (L.) Murr.
Dao Thi Hai Ly (*), Nguyin Viel Thang (**), Mai Sy Tuan i*"}
Khd ndng diit bg gdy mudi Anopheles, Culex vd Aedes cua dfch chiet ethanol md seo cdy cue do Spilanthes acmella (L.) Murr. duac nghien cdu. Dich chiet dc chi' mgnh nhdt vdi cdc lodi muoi Anopheles vdi cdc gid trf LD50 tuang iing (d 0,46; 0.59; 0,74 mg/ml cho Anopheles episolicus.
Anopheles minimus. Anopheles dirus. Mudi Culex quinquesfasticus it bi diit nhdt trong cdc lodi dd tha (LD50=2,79mg/ml) roi di'n Aedes aegypti (LD50=l,65mg/ml). Ket qud con cho thdy dich chidt lie che tdt vdi mudi lUnhien (LD50=l,29mg/ml). Tdc dgng cua dich chiet cung ldm bo gdy keo ddi thdi gian sinh trudng vd chdm chuyen sang qudng.
TU khoa: md sea Spilanthes acmella. Anopheles episoticus. Anopheles minimus. Anopheles dirus.
Culex quinquesfasticus, LD50
Mosquito larvicidal activity of spilanthes acmella (L.) Murr. callus extract
Dao Thi Hai Ly (*>, Nguyen Viet Thang (*•), Mai Sy Tuan {***)
Larvicidal activity ofeihanolic extract of Spilanthes acmella callus against Anopheles, Culex and Aedes larvae was studied. The extract was the most effective on Anopheles with LD50 values al 0.46;
0.59; 0.74 mg/ml respectively on Anopheles episoticus. Anopheles minimus. Anopheles dirus. Aedes aegypti larva was less susceptible lo the callus extract after Culex quinquesfasticus as revealed from LD50 values (at l,65mg/ml and 2,79mg/ml). The results also show that the extract had a larvicidal effect against the mosquitoes collected from natural environment (LD50=],29mg/ml). In addition, there was a delay in the development of larvae to the pupal stage when the larvae were exposed to the extract.
Key words: Spilanthes acmella (L.) Murr. callus. Anopheles episoticus. Anopheles minimus.
Anopheles dirus. Culex quinquesfasticus. LD50
Tap chl Y td Cdng cpng, 7.2010, Sd 15 (15) 29
I Td'HG QUAN & NGHIEN Cliu I
T^c gia:
Ths. Dio Thi Hai Ly - Tnrdng Dai bpe Stf pham Hi NOi. Dien thoai: 0975652635.
E.mail: [email protected]
NguySn Vidt Thing - Hgc vien cao hoc Khoa 17 Khoa Sinh hoc, TnTdng Dai hoc Stf pham Hi Ngi.
Dia cW: 136 XuSn TbiSy, Cau GiTy, Hi Npi. Dien thoai: 0912212149. E.maU: [email protected]
^,**, PGS.TS. Mai Sy Tuan - Khoa Sinh hpe, Tnrdng Dai hoc Srf ph?m Hi Ngi, 136 Xuan Thdy. Clu Gia'y, Ha N$i.
Di^n ±oai: 0913017097. E.mail: [email protected]
{ * )
(•')
1. D$t vSh d l
Muoi Id vecld trong gian truyin nhilu bfinh nguy hiem cho ngffdi. Kiem sodt cdc vectd nay dang dffng trffde thdch Ihffc vi kha nang khdng ciia mufii vdi cdc Ihudc trff cdn iriing hda hoe truyin Ihd'ng.
Mat khdc, cde thudc ndy cd the anh hffdng ddn cde sinh vai cd Idi, dpng vat va ca con ngffdi. Thudc Irff cdn triing ed nguin gde thffc vat Id mpl hffdng Ihay Ihe tilm ndng vi nd an loan, de phan hiiy vd cd the kidm b khip ndi trfin the gidi. Cay ciic do Spilanthes acmella (L.) Murr. Id mpl trong nhffng cay co cd boat tinh difil cdn trung (Pitasawat et al., 1998), va ca bp gay mu5i (Naih et al., 2006). Vipc nhan gifing bang nudi cay md va thu chdt chifi't tff mfi seo ciia eay cdc do Spilanthes acmella (L.) Murr. da dffde tien hanh Ihdnh cdng (Mai Sy Tuan et al., 2009).
Trong bdo cdo nay, ehffng tdi lap trang nghifin cffu kha nang ffc chd va diet bp gay muoi cua dich ehiet md seo eay eiie do vdi hi vpng cd the tao dffde chat difit mu6i bing ffng dung cdc ldi ihe ciia ky thuat nuoi ca'y md thffc vdt.
2. PhUdng ph^p nghifin cffu 2.1. Au triing mudi
Cdc loai bo gay mudi Anopheles minimus.
Anopheles diras, Anopheles episoticus, Aedes aegypti, Culex quinquefasciams mua tff 'Vifin Sd't rdt- Ky sinh iriing - Cdn iriing Trung ffdng d eac dp tudi 1, 2, 3 dem ve nudi tai phong thi nghifim dfi'n khi dat tu^i 3 Ihl dem ihff. Phffdng phdp nudi: lay nffdc mdy vdo khay dai mffe nffde 2-2,5em, tha bp gay muoi vdi mat dfi < l-2con/cm', cho dn vdo sdng sdm bang thffc dn nudi meo vdi Iffdng Img/khay 150 - 200 bp gay/ ngay.
Au Iriing mudi tff nhifin dffde bay bang cdch dat chdu nffde d vffdn Ihffe nghifim dfi'n khi xud't hifin bp gay tudi 1 thi bit dau cho dn, nudi den luoi 3 Ihi thu
lam Ihl nghipm.
3 Ioai muoi Anopheles d trfin Id vecld trayin bfinh sd't rdt, mudi Aedes Id vecld truyin bfinh sdt xua^t huyet vd Culex quinquefasciams truyin bfinh giun chi baeh huyfi't.
2.2. Chudn bi dich chiet md Sfo Cdy cue do dffde thu thdp, khff trCing vd nudi lao md seo nhff bdo edo trffde da trinh bay (Mai Sy Tuan et al., 2009). Nguyfin lieu thffc vat dffde khff triing bing 0,1% HgC12 trong 10 phdt va cay Ien moi trffdng lao chdi MS -I- 0,5 mg/I BAP + 0,1 mg/1 kinetin ->f 30g/I saecarose + 8g/l agar. Than, Id cua chdi in vitro dffde ea'y ifin mdi trffdng tao md seo ed Ihdnh phin Id MS + 0,5mg/l 2,4D -(- 0,5mg/I BAP + 30g/I saecarose + 8g/I agar, nudi d dilu kipn ldi, nhifit dd 25 ± 20C. Md seo sau khi hinh thdnh dffde ca'y nhan irfin mdi trtfdng cilng thdnh phan de dai khdi Iffdng can thidt.
Chifi't nit md seo thu dich ihd nhff sau: Can 50g md seo dem ngam irong lOOml eon irong 24 gid, ly ldm thu dich noi Ian 1, can dtfdc ngam tifi'p trong lOOml edn vd thu dich lan 2, lap Iai bffdc nay den Ilic dieh ehiet khdng mdu (ihffdng 5 Ian). Td'i ca dich dffde irdn lan vdi nhau, dem cd bang mdy cd chan khong ihu can chiet. Hda lan can chiel trong nffdc cat de Ihu dich ehiet thd dang dung dieh me.
2.3. Thd khd ndng dc chevd diit hg ggy mudi (WHO, 1981)
Kha ndng ffc chd va diet bp gdy muSi dffde tien hanh nhff phffdng phap chuin ciia WHO. 25-30 bp gdy muSi dffde tha vao mdi cdc dffng 250ml nffdc mdy, bd sung dung dich me cua chat ihff de cde cdc dat ndng dp khac nhau. Nudi mu6i d nhifit dp thffdng, sau 24h quan sal ddm sd muoi chdt. Ddi ehffng khdng bd sung dieh chiel.
Sd lieu dtfdc phdn tich bang phan mim 30 Tap chi Y te Cdng cdng, 7.2010, Sd 15 (15)
ITflKHUtllGmEIICIIIII
MINITAB 15 d« linh cdc giS tri LD50. gidi hao tren ligidi lian dirdi d klioang tin cay 95%, d^ danh gia miJc dO tac dgng ciJa chfl thiir vdi cac loai mnoi.
3. KSt qua nghiin cthi
Thi nghidm dau ti6n s i dung chi't diij cd nong doj), 1 mg/ml -1 mg/ml tren muoi Aedes aegypti cho thay Ithong c6 bo gjy nao bi ehft sau 24 gid thi nghifm. Tuy nhien, Ithi nu6i th«m thdi gian thi sir sinh tnrdng cila bo gSy giam din tff cac 16 cd nSng dd cao den cScId cd ning do thfp. Ngoii sU khac nhau v l kich thirdc, d cic 16 thi nghifm cdn cd sil khac nhau ve su chuyin giai doan trong qua trinh phiSt triin Clia bo gly. Vdng ddi mudi gom 4 giai doan 14 tntng, bo gjy, quang v4 trirdng thanh. Khi bl xil I^ a ndng do cao thi sd bo gay chuyen dirdc sang giai doan qu5ng rat thap, d nong do 1 mg/ml, chi 24% sd quing tao ra sau 72 gid trong khi d nong dd 0,lmg/ml ti W nay li 44% vi sau 84 giS thi chi 40% quang xuat hipn d ndng dp Img/ml (bang 1).
Bdng 1. Anh hudng cua dich chiSi mo sfo cay ciic do len ftp g4y Aedes aegypti
ThM gian
72h 84h
Ndng dO c}n chift (mg/ml) 0 0,1 0,3 0,5 0,7 1
% quing tao thinh 44
72 44 72
44 60
44 56
32 52
24 40 Dieh ehifi'l nghifin cffu cd idc ddng manh den bp gdy cua edc Iodi Anopheles. An triing Anopheles minimus. Anopheles episolicus deu che'l 100% sau 14 gid xff Iy d ndng dp l,2mg/ml con Anopheles dirus Id d l,5mg/ml (bang 2).
Bdng 2. Anh hUdng cua dich chiei mo sfo cdy cue do den 3 lodi Anopheles
Lofimndl
Anopheles Anopheles dirus Anopheles maoncus
Ndag Alt C4D cblCt (me/ml)
DOI 0.05 IOI 1 0.3 j o.s 1 o? lo9[ 1.2 1 u
» d i f i
0 0 0
0 0
"
8
"
12 12 S 28
36 S 40
48 16 56
64 S4 80
100 96 100
100 100 100
Trong khi dd so' lieu 5 bang 3 cho tha'y ca 2 Ioai Aedes aegypti, Culex quinquefasciams va ca mudi m nhien deu II mdn cam vdi dieh ehiet hdn so vdi cdc loai Anopheles, de diet 100% bp gay ein tang ndng dp Ifin 7,5-lOmg/ml.
Bdng 3. Anh hudng cua dich chiet mo seo cdy cUc do den Aedes aegypti, Cidex qidnquefasciatus vd muoi tu nhien
—
AiJaara^
Cmla
H a & V a J b h i
N I 4 d4 I 4 D cbVl (m^Dl)
. 1 • 1 ,. . 1 " 1 . 1 s 1 " .0 U
« d i 6 0
•
0
»
46 67 S34
,.
53 33 60
36.61
SJ33 60
«
66 67 733
60
73 33 90
76.67
an 100
7&67
S6 67 100
so 90
100
.,»
100
Khi xff Iy sd'hfiu va tinh todn, gia tri LD50 eiia eac loai mudi la 0,46; 0,59; 0,74; 1,29; 1,65; 2,79 mg/ml (bang 4) ttfdng ffng vdi mtfc dd tdc ddng tff manh ddn ydu cua dieh chidt md seo vdi cdc Ioai muoi Anopheles episoticus. Anopheles minimus.
Anopheles dirus, mudi Iff nhien, Aedes aegypti, Culex quinquesfasticus.
Bdng 4. Gid trj LDSO vd khodng gidi han cua djch chie't vdi cdc logi bg ggy mudi
Lo»i bQ gjy muai
AnofAetes episoticus Anopheles imniauu Anoplteles dtnii MnSinrohicEi Aedes aegypti Culex tpiaiguesfasticus
LDSO (mgADl)
0 4 6 0 J 9 0 7 4 1.29 1.65 2.79
95% CI Giiti h^o duA
0 36 0.48 0.66 0.73 1.15 2 1 9
Gidl h ^ tt&u 0.567
0.72 0.B1 1.77 2.00 3.41
4. Ban luan
Promsiii va ddng lae gia (2006) cfing cho bidl dich chifi't cua 3 loai eay Mammea siamensis, Anelhum graveolens, Annona murieata lam klo dai thdi gian sinh trffdng va phat trien cua bp gay Aedes aegypti va ffc chd sff xult hifin mudi trtfdng thdnh, nguyfin nhdn cd the do Iffdng hoocmon duy tri trang thai chffa trffdng thanh trong bp gay eao hoac do tac ddng cua cdc hdp chd^t trong cdy. Zebritz (1984) khi nghifin cffu dich chiet hat Azadirachta indica I6n bp ga\ Aedes aegypti cho rang azadirachtin Id nhan td lam khang hoocmon ecdysieroid do dd sff sinh trffdng hi ffc chdva sff phat men bi keo ddi.
Tap chi Y te Cdng cpng, 7.2010, Sd 15 (15) 31
[ T A K G QUAN & NGHIEN CUOI
Nghien cffu v e tdc dung cua dich chiet e a y cue d o Spilanthes acmella (L.) Murr. Ifin bp g a y mudi da dffde mpt s d tdc gia b a o c a o nhff Saraf va ddng tae gia (2002) cho rang Spilanihol trong dieh chiet ethanol ciia hoa cue do cd dac tinh diet trffng, bp g a y va q u a n g ciia mudi A n o p h e l e s cuHcifacies, Culex quinquefaeiams va A e d e s aegypti Linn.
Nalh vd dong tdc gia (2006) cung bdo cdo kha nang diet Culex quinquefasciams, A e d e s albopicms cua dich chiel Id Spilantiies acmella vdi gid tri LC50 ttfdng ffng la 57,4 vd 54, I p p m . Cao hdn mpt it la gia tri LC50 bang 61,43ppm cua dich chiet cay nay vdi b p gdy Culex quinquefaseiatus do Pitasawat va ddng Idc gia (1998) bdo cdo. Tuy nhifin, chiing tfii chffa thay bdo cdo ndo ve dieb ehidl md seo cay cue do len bp gay mudi. Khi Ihff dinh linh vdi thudc thff Dragendorf, M a y e r thi khdng phdt hidn thdy alkaloid trong dich ehidl ehffng id, md seo khdng cd spilanihol (la 1 alkaloid). Vdy trong dich chidt md
s e o cdn cd chi't khae n g o a i spilanihol cung cd kha ndng ffc c h d cdn triing ciia eae loai muoi Anopheles, C u l e x va A e d e s .
Kdt qua ndy eho thay dich chidt md seo cay ciic a o cd kha ndng ffc c h d lot c a c loai b o gdy mudi nhat Id m u d i A n o p h e l e s v d i gid tri L D 5 0 < l m g / m I . Nhff vay l a o c h d p h a m diet m u d i tff ihffc vdl nhd tfng dung ky ihudi nudi c a y m o i d bdo id hffdng kha thi vl ffu d i e m chu dpng dffde nguydn lieu, d i l u kien nudi cl^y ciia ky thuat n a y vd tinh chl^l thdn ihifin mdi trffdng c u a hdp chat sinh hoc tff ihffc vat. Vipc nudi cdy mo seo trong m d i trtfdng long dang dffde nghifin cffu d e n a n g quy md s i n xui't Ien ldn hdn d mffe edng ngbipp trong c a c binh Ifin men.
Ldi c a m d n :
Kinh phi thffc hifin tff d e tai c a p E B2008-17-119TD
i trpng diem
Tdi li$u tham khao
Ti^ng Vtet
1. Mai sy Tulfn, Mai Tbi Haag, Dio Tbi Sen, Dio Tfai Hai Ly (2009). Hoat tinh khang Idiuan va ffc che t£' bdo cda dich chidt md S90 cSy ciic io boa ving SpUantbes acmella (L.) Murr. Ky yg'u H^i nghi C6ng nghfi sinh hpe toln qudc 2009:
447-451.
Tie'ng Anfa
2. Nath DR. Bbuyan M, Goswami S. (2006). Botanicals as MosQuito Larvicides. Defence Science Joumal 56 (4), 507- 511.
3. Pitasawat B, Choochote W, Kanjanapothi D, Panthong A, Jitpakdi A, Chaithong U. (1998). Screenmg for larvicidal activity often carminative plants. Southeast. Asian J. Trop.
Med. Public Health 29 (3): 660-662.
4. Promsiri S, Naksathit A, Kniatrachue M, Thavara U, (2006). Evaluations of larvicidal activity of medicinal plant extracts to Aedes aegypti (Diptera: CuUcidae) and other effects on a Don target fish. Insect Sci 13: 179-188.
5. Saraf DK, Dixit VK, (2002). Spilanthes acmella Murr Study on its extract spilantho] as larvicidal compound. Asian L Exp. Sci 16: 9-19.
6. Zebitz CPW, (1984). Effects of some cmde and azadtrachtin-enriched neem (Azadirachta indica) seed kernel extracts on larvae of Aedes aegypti. Aosualian Joumal of Entomology 39: 208-211.
7. Wodd Health Organizauon (1981). Instructions for determining tbe suscepubili^ or resistance of mosquito larvae to insect development inhibitor. WHO/Vector Biology and Connol: 812-881.
32 Tap chi Y td Cdng cdng, 7.2010, Sd 15 (15)
