86 TAP CHl PH6NGCH0NG BgNH SOT RET VAcAC BgNH KYSINHTRUNG S6 1-2012
XAC DINH Mtrc NHAY CAM V 6 I HOA CHAT Dlf T C 6 N TRUNG CV A ANOPHELES MINIMUS CHUNG P H 6 N G THI NGHIf M VA
THVC DJA TINH HOA BINH BANG PHUONG PHAP T H U SINH HQCvA HOA SINH (GIAI DOAN 2009-2011).
Bio Minh Trang, H6 Dlnh Trung Vien S6t rit-KST-CT TU' Tdm tit •
Nghifn ctiu tiin hdnh tit ndm 2009 din ndm 2011 v&i 5 qudn thi Anopheles minimus hoang dgi thu Ihdp tgi thifc dia tinh Hoa Binh, vd 2 chiing Anminmus nudi trong nhd nudi cdn triing. Trong sd 5 qudn thi An.minimus hoang dgi cd 4 qudnthf nhgy cdm v&i Lambda- cyhalothrin (ty If mudi chit 98%) vd Alpha-cypermlhrin (ly If mudi chit 100%), mgt quan thf nhgy cdm vcri Alpha-cypermethrin (ty If mudi chit 100%)^ vd co khd ndng khdng viM Liimbda- cyhalothrin (ty If mudi chii 96%). Hdu hit cdc qudn Ihi nghifn ciiu deu co ty If chft trong tha sinh hgc & thi hf Fl thdp hgn so v&i mudi hoang dgi tgi thuc dia. Chi cd qudnjhe An.
minimus Rdy (Vdn San, Lgc San, Hda Binh) cd ty If chit Irong thit sinh hgc a muSi hoang dgi vd mudi Fl luang duang nhau (98% - 100%). An. minimus Hda Son (chung phdng thi nghifm) khdng v&i hai hda chdt trong hai ldn Ihii (ly If mudi chit Irong ldn thit Ihii nhdt la 62%, ldn thii thii hai la 74%). An. minimus Quyit Chiin (chimg phdng thi nghifm) nhgy v&i hai hda chdt Ihir (ty If chit 99-100%) trong Ihu sinh hgc ndm 2009, co khd ndng khdng v&i hai hda chdt thii trong ndm 2010 (ty If mudi chit 82-85%)._Hogt linh enzym esterase vd monooxygenase cd xu hu&ng tdng & cdc qudn the cd ty If muoi chft thdp han trong thir sinh hgc, thf hien ro & mudi Fl.
1. DAT V A N B E
Hifn nay, Vift Nam cung nhu nhilu nude trSn thi gidi, chuong trinh phdng chong sit ret (PCSR) dang su dung mpt so hod chit nhdm pyrethroid dl phdng chong vec-to, nhu Alpha-cypermethrin, Lambda-cyhalothrin, Permethrin. Cdc hod chdt ndy dupe dimg df phun tin luu, tim mdn, ldm huong xua. cho hifu qud khd tit. Tuy nhiSn, sau nhilu ndm sd dyng rpng rdi cdc hod chit dift cdn trimg, vife phdt ttiln tinh khdng cua cdc vfc-to truyin bfnh Id khd ttdnh khdi, dan din hifu qud cda cdc bifn phdp phdng chong vec-to sit tdt bdng hda chit cd thi bi gidm thip hodc thdm chi mit tdc dyng.
Trong so khdng nhilu cdc phuong phdp xdc dinh tinh khdng hod chat dift cdn trung d cdc lodi vf c-to sdt rft, till phucmg phdp Thd Sinh hpc (bioassays) vd Phuong phdp Hod Smh (Biochemical analysis) Id 2 phuang phdp dupe sd dung nhilu nhit. 6 Viet Nam, xdc dinh tinh khdng cua muoi vdi hda chit ehd yiu bdng Thd Sinh hpc t^ thyc dia. Phuong phap nay cd uu diSm Id don gidn, de thyc hifn, dd tin c^y cao, nhung nhupc diSm Id phdi phan tich mft sd lupng muli tuong ddi ldn vd khdng xdc dinh dupe co chi khdng dl dua ra gidi phdp dng phd thich hpp. Vi v^y, dl fliu dupe thdng tin diy du hon vl tinh khdng hod chit, can tiln hdnh ddng thiri phuong phdp Thu Sinh hpc vdd mpt phuong phdp khdc, chdng hgn vdi phuong phdp Hod Sinh nghiSn cuu boat tinh cda mdt sd enzyme liSn quan din cdc co chi khang khdc nhgu.
Trong boi cdnh dd, vife triln khai thyc hifn dl tdi "Xdc dinh muc nhgy cdm v&i hod chdt dift cdn triing cua Anopheles minimus chiing phdng thi nghifm vd thuc dia tinh Hod Binh bdng phuongphdp thit sinh hgc vd hod sinh (giai doan 2009-2011)" Id rdt cdn thilt, vdi hai mye tieu sau day:
S61-2012 TAP CHl PHbNG C H 6 N G BgNH S 6 T RET VA CAC BgNH KY SINH TRCING 87
(I). Ddnh gid dp nhgy cam vdi Lambda-cyhalothrin vd Alpha-cypermethrin cda An.
minimus chirng phdng thi nghifm bdng phuong phdp Thu Sinh hpc vd Hod Sinh phyc vy chpn Ipc ddng muli nhay de nudi gid chiing.
(2). Ddnh gid dp nhay cam vdi Lambda-cyhalothrin vd Alpha-cypermethrin cda An.
minimus d mpt so dja phuong tinh Hod Binh bdng phuong phdp Thd Sinh hpc vd Hod Sinh nhdm xdc dinh nhihig qudn thi muli nhgy cd the sd dyng trong nghiSn cim vd ddnh gid hifu lyc cda hod chit dift cdn trimg
2. SJA DiftM, T H 6 I GIAN, B 6 I TU'giNG, VAT LI^U, PHUONG PHAP NGHlftN CtTU 2.1. Dja diem, thdi gian nghifn cuu
- Dia diem: Thyc dia: Tinh Hod Binh; Phdng tiii nghifm: Vifn SSt rdt-KST-CT TU Thdi gian: Td tiidng 6/2009 din 6/2011
2.2. Dot tupng nghien cuu
- Muoi cdi An. minimus chdng phdng thi nghifm:
+ Chung An. minimus Quylt Chiln + Chtmg An. minimus Hod Son
- Mudi cdi An. minimus tiiu fligp tai 5 diim tiiyc dia thudc thih Hod Binh:
+ Thdn Trung Hung, xd Quylt Chiln, huyfn Tan Lgc + Thdn Khdi, xa Phd Cucmg, huyfn Tdn Lgc + Thdn Ba Rd, xd Nat Son, huyfn Kim Bdi + Thdn Chum, xd My Thanh, huyfn Lgc Son -^ Thdn Rdy, xd Vdn Son, huyfn Lgc Son - Hai enzym phan tich la: esterase vd monooxygenase
- Hai hod chit thu nhay cam theo phuong phap Thd Sinh hpc (bioassay):
Alphacypermethrin 30mg/m^ vd Lambda-cyhalothrin 0,05%
3.3. Vdt lifu:
- Dyng cy hod chit cho thd sinh hpc:
+ BS dyng cy thd nhgy cam theo tiSu chuin cua Td chdc Y tS the gidi + Gidy tim hod chit dift cdn triing vd giiy dli chdng (Malaysia sdn xuat).
- Dyng cy hod chit cho phan tich hod sinh:
+ Mdy dpc ELISA vdi hf tiling phan tich ELISA cua Thermo
+ Gilng nhya day bdng 96 gilng, pipet, may Iy tam eppendorf Ignh, ta Ignh tiiudng, + Td Ignh sdu -20°C & 70°C, mdy do pH, mdy khuiy td, cdn difn td....
3.4. Phuong phdp nghien cuTi 3.4.1. Phinmg phap thu mau t?i thuc dia
Muli Anopheles minimus cdi dupe tiiu thdp bdng phuong phdp soi bdt mudi dchudng gia sdc ban dfm vd mdi ngudi bit muli ban dSm tiieo qm ttinh cua Vien sot ret-KST-CT TU Mdt phin muli thu tiiap dupe dung cho tiiu sinh hpc tgi tiiyc dia xac dinh dp nhgy cam, mpt
88 TAP CHl PHtiNGCHONG BgNH S(iT RgT VACAC BgNH Ky SINH T R Q N G Sti 1-2012
phan dupe bdo qudn trong nito ldng dl phyc vy cho cdc phdn tich hod sinh tai phdng til!
nghifm xdc djnh hogt tinh enzym.
3.4.2. Phuong phdp thd- Sinh hpc (Bioassay)
Phuong phdp thd sinh hpc dupe thyc hifn theo qui trinh cua Tl chdc Y te till gidi (WHO/CDS/CPC/MAL/98.12.) [6]
Ddnh gid kit qud mdc dp nhgy cdm theo tiSu chuin cua WHO (WHO/CDS/CPC/MAL/98.12) - Ty If muli chit 98-100%: muoi nhgy cdm
- Ty If muoi chit 80-97%.: muli cd khd ndng khdng hod chit - Ty If muoi chit <80%: muli khdng hod chit
3.4.3. Phuong phdp hod sinh (Biochemical assay)
Phucmg phdp hod sinh dupe thyc hifn tiieo quy trinh cua Tl chdc Y te till gidi (WHO/CDS/CPC/MAL/98.6.) [5]
Ddnh gid dd nhgy cdm bdng thd hod sinh - Esterase
Hogt dp cug EST theo phuang phdp dpc lien tyc su dyng ca chit PNPA dupe tinh theo cdng thirc sau:
EST=—9P5^—(nmol/phdt/mg protein) [3,5]
lP]x\ 3,3x0,6
Nlu boat dO cda esterase vdi vife su dyng co chit PNPA > 0,07387pmol/phdt/mg protein cd the bieu hifn khdng [4].
- Monooxygenase
NSu boat df P450 > 0,9nmol/mg protein: cd thS bifu hifn Ichdng [4].
3.5. Xir Iy so lifu: Trong nghien cim boat tinh enzym esterase vd monooxygenase, gia tti hogt dp tnmg binh dupe so sdnh bdng t-test.
3. KtT QUA vA B A N LUAN 3.1. Ket qua thir sinh hpc
Ket qua thir sinh hpe doi vdi muoi thu thap vd Fl t^i th^c dfa Hod Binh.
Trong giai doan 2009-2011, cd tit cd 5 quin thi An. minimus tai Hod Binh dupe tiln hdnh ddnh gid muc nhgy cdm vdi hai hod chit dift cdn trimg Id Lambdacyhalothrin 0,05% vd Aphacypemnethrin 30mg/m^.
Trong 5 qudn thi nghifn cim cd duy nhit mpt quin thi cho kit qud cd khd ndng khdng vdi lambdacyhalothrin (ty If muSi chit taong ling 96%), cdc qudn thS cdn Igi nhgy vdi cd hai hod chit thd (bdng 1). Kit qud ndy tuong ty vdi cdc kit qud cua Vfl Due Chinh 2004 vd Nguyin Thi Bf ndm 2009 [1,2]. Cd till tiiiy rdng it cd sy biln dpng mdc dp nhgy cam cua cdc quin till An. minimus tai Hod Binh trong vdi ndm trd Igi ddy. Dieu ndy cd thi do tai day hiu nhu dd ngimg sd dyng hod chit dift cdn trimg ttong phdng ching sdt ret.
Trong 5 quan thi nghien cdu cd mpt quan thi cho kit qua thd nhgy cdm muoi Fl (bdng I) Id nhgy vdi cd hgi hod chit trong cd hai ldn I|p Igi thd nghifm (vdi ty If muli chit taong img vdi cd hai hod chit tu 98-100%) Id muli FI cda quin thf tgi Rdy; mot quin thi cho kit qud thd nhgy cam muoi FI Id khdng vdi cd hai hod chit (vdi ty If muSi chet tuong irng 66-75%)) la muoi FI cda quin thS tgi Bd Rd; ba quin thi cdn lgi cho kit qua thd nhgy cdm mudi FI Id cd khd ndng khdng vdi hod chit (vdi ty If muoi chit, tuong dng vdi hai hod chit ta 82-96%)
S61-2012 TAP CHl P H 6 N G C H 6 N G B $ N H S 6 T RfeT VA CAC BgNH KY SINH T R O N G 89
Bing 1. K^t quS thijt sinh hgc xdc djnh mirc nhay cam ciia muoi An. minimus nam 2009-2011.
Dia diem
ThfinTnuig Hng, xa Quy^t Chien, huyin TSnL?c Th6n Khdi, xa Pha Cutog, huyfn Tfin Lac
Th6n Ba Ra, xaN$tS(m, huyfn Kim
B6i Th6n Chum, xS My Thanh, huyfn Lfic Son
Thon Rdy, xa van Son, huyfn L?ic Son
Phan loai
S6 mu5i thir S6 mu6i ch^t Ty If ch^t S6 muSi thu S6 mu6i ch^t Tylfch^l S6 mu6i thir S6 muoi ch^t Ty If chit S6 mudi thu S6 muSi ch^t Ty If chSt S6 mu6i thir So muoi chet Ty If chft
Nam 2009 Lambda- cyhalothrin
0,05%
NC 99 97 98 78 77 98,7 100 99 99
Fl 100
82 82 97 89 92 89 59 66
Apha- cyperrmethrin
30mg/m2 NC
100 100 100 79 78 98,7 102 102 100
Fl 99 92 93 94 90 96 96 72 75
Nam 2010-2011' Lambda- cyhalothrin
0,05%
NC
96 92 96 97 97 100
Fl
114 106 93 100 100 100
Apha- cypemnethrin
30mg/m2 NC
100 100 100 100 98 98
Fl
102 93 91 101 (100)
100 (99) 100 (99)
D6i chiing
0
0
0
0
0 Ket qua thu sinh hoc mu5i An. minimus a the he Fl cua cac quan the co ty le chet thap han so voi cac quin thi bit tgi thuc dia. Diiu niy c6 thd do muoi Fl dong deu va cung liia tu6i hcfn so vai mu5i bat tai thuc dia.
Ket qua thu- sinh hgc doi voi chung muoi phdng thi nghiem
B^g 2. K6t quk thi sinh hpc xic djnh miic nhay cam ciia mu6i An. minimus PTN n5m 2009-2011
Dja diim
Chiing v4». minimus Quyit Chiin
Chiing An minimus Hoa Son
S6 mu5i thu S6 mu6i chit Tylf chit (%) So muoi thir S6 muoi chit Tylf chit (%)
Nam 2009 Lambda- cyhalothrin
0,05%
100 99 99 100 62 62
Apha- cypemnethrin 100 100 100 103 72 70
Nam 2010 Lambda- . . cyhalothrin ^ , .
0,05% 1 <^5T=rrmethnn 100 96 85
85 102 64 63
79 82 99 73 74
Dii chdng
0
0 Trong cd hai lin thd nhgy cdm trong hai ndm 2009 vd 2010 (bang 2), chung An.
minimus Hod Son cho kit qud khdng vdi hai hod chit lambdacyhalothrin vd aphacyperrmethrin (vdi ty If muoi chit tuong dng 62-74%), chdng An minimus Quylt chiln
90 TAP CHl P H 6 N G C H O N G BgNH SOT R 6 T VA CAC BgNH KJ SINH T R O N G S 6 1-2012
tai thdi diim thd ndm 2009 cho kit qud nhgy vdi cd hai hod chit vd ndm 2010 chung ndy cho kit qud cd khd ndng khdng vdi hai hod chit vdi tj' If muli chet tuong dng trong thd sinh hpc Id 82-85%)
3.2. Kit qud phdn tich hod sinh
3.2.1. Kit qud phdn tfch hoyt d$ Monooxygenase
Theo kit qud (bdng 3) hogt df monooxygenase trung binh cua tit cd cdc quan thi diu
<0,9nmol/mgprotein, cdc quin thi diu nhgy cdm vdi vdi cdc hod chit dift cdn trung thudc nhdm pyrethroid. Kit qud ndy khd tucmg ddng vdi kit qud thd sinh hpc tgi thvrc dia, liau het cdc qudn thi ndy deu nhgy cdm vdi hai hod chit Id lambdacyhalothrin vd alphacypermethiin (vdi ty If chit tuong ung trong thd sinh hpc 96-100%).
Dl so sdnh tuong quan hogt dO gida cdc quin the chdng tdi chpn mdt quin thi cd kit qud nhgy cdm vdi hai hod chit dift d cd muoi thvc dia vd muli Fl Idm doi chdng. Dd Id quin the muoi tgi thdn Rdy. Ket qud cho thiy cd s\f tdng hogt tinh cda cdc quan thi cdn Igi so vdi quin thi ndy.
Bdng 3. Kit qud phdn tfch hogt df monooxygenase d quin thf An. minimus thuc dia Hod Binh
Quin The Khdi, PC, TL Ba Ra, NS, KB Trung Hung, QC, TL Ray, VS, LS Chum, MT, LS
Hogt df trung binh (nmol/mg protein)
0,58 0,66 0.32 0,22 0,31
Df tin cfy (95%)
0,07 0,08 0,05 0,042 0,047
Khodng btenthien GTTBvdi do tin
cfy 95%
0,51-0,65 0,58-0,74 0,27-0,37 0,18-0,26 0,27-0,37
Gid tri p so vdi quan the Rdy,
VS,LS 0,00142 t 5,43E-05 T 0,002442 T
0,005221 T Ben canh vi^c xdc djnh ho^t dp enzyme monooxygenase trung binh quan the, ty I^
phin t r ^ so cd the cua quan thS c6 gid trj ho^t d$ theo cdc khodng khdc nhau cung dupe dua ra. Ket qua du<?c trinh bay trong bang 5. Theo d6, c6 hai quAn the la quan the muoi An.
minimus tm Khin vd Bd Ra c6 ty 1? nhit djnh so cd thi vugt nguong khdng. C\i thi Id 8,5%
s6 cd thi cua quiin thi tai Khdi vd 15,2 % so cd thi cua qudn thi tai Bd Ra vupt ngu5ng khdng.
Bdng 4. Ty \% % so ck thi quin thi phdn theo cdc khodng gid trj hofit d$ monooxygenase
Quan the ^ \ ^ Khdi, PC, TL
BdRd,NS,KB Trung Hung, QC, TL Rdy, VS, LS Chum, MT, LS
Gid tri hogt dp monooxygenase theo cdc khodng (nmol/mg protein)
<0,1
0,0 0,0 0,0 25,1 2,1
0,1- 0,2 0,0 3,0 24,2 7,0 25,5
0,2- 0,3 0,0 6,1 30,3 27,9 27,7
0,3- 0,4 19,1 12,1 36,4 14,8 23,4
0,4- 0,5 17,0 18,2 3,0 8,2 12,8
0,5- 0,6 23,4 18,2 3,0 5,0 2,1
0,6- 0,7 19,1
6,1 3,0 9,1 0,0
0,7- 0,8 8,5 12,1 0,0 0,0 2,1
0,8- 0,9 4,3 9,1 0,0 2,9 4,3
0,9- 1,0 6,4 6,1 0,0 0,0 0,0
>1
2,1 9,1 0,0 0,0 0,0
S6 1-2012 TAP CHl PH6NG CHONG BgNH S 6 T RgT VA CAC BgNH KY SINH TRONG 91
Kit qud phdn tich hogt dp monooxygenase & mudi Fl.
Kit qud bdng 5 cho thiy tit cd muli Fl cda cdc quin the diu cho hogt dp monooxygenase nhd ban 0.9nmol/mgprotein. Nhu vdy theo ngudng nay tit cd cdc quin thi FI nhgj; cdm vdi hod chit nhdm pyrethroid, nhung theo kit qud thd sinh hpc, hiu hit cdc quin thi Id khdng hodc cd khd ndng khdng vdi hai hod chit thd, chi cd duy nhit mOt quin thf tgi thdn Rdy cho kit qud nhgy cdm vdi cd hgi hod chit. Tuy vgy, cung nhdn thiy cd su tdng boat dp mpnooxygenase d the hf FI cda cdc quin thi nghifn cdu so vdi muoi Fl cda quin thf d Rdy. Ndi cdch khdc cd su tdng hogt tinh monooxygenase d cdc quin thi cd ty If chit thip hon trong thd sinh hpc.
Bdng 5. Ket qua phan tich hogt dp monooxygenase d till hfFl cua cdc quMthe An.minimus thuc dja Hod Binh
Quan The Khdi, PC, TL BaRa,NS,KB Tmng Hung, QC, TL Ray, VS, LS Chum, MT, LS
Hogt df trung binh (nmol/mg protein)
0,36 0,42 0,38 0,19 0,181
Cl (95%)
0,07 0,067 0,038 0,027 0,030
Khodng biin thien GTTB. Cl 95%
0,31-0,42 0,35-0,49 0,34-0,42 0,16-0,22 0,16-0,21
Gid trj p so vdi quan thf Rdy, VS, LS
0,00355lt 0.004294 t 0.002327 t 0,751716 Gia tri ho^t dp monooxygenase a Fl ciia cdc quan the cung dugc phan tich theo cac khodng gid tri hoat dp khac nhau.
Theo kit qua (bdng 6) c6 3 trong 5 qudn thi co ty le nhdt dinh s6 ca thi vupt nguSng khdng Id:
Thi he Fl cua quin thi Kh6i: 3,4%, quin thi Ba Ra: 2,7% vd qudn thi Trung Hung: 1,4%
Bang 6. Ty 1? % s6 ca the quin thi phan theo cac khodng gia tri hoat dO monooxygenase.
Qudn thi ^ ^ ^ ^ ^ KhdiFl,PC,TL B a R d F l , N S , K B Tning Hung F1,QC,TL R4yFl,VS,LS Chum, MT, LS
Gia tri boat df monooxygenase theo cdc khoang (nmol/mgprotein)
<0,1 6,9 0,0 2,9 14,0 25.5
0,1- 10,2- 0,2 0,3
17,2 27,6 5,4 13,5 7,2 31,7 42.6
29,0 15,0 14.9
0,3- 0,4
13,8 35,1 30,4 25,3 12.8
0,4- 0,5- 0,5 0,6
6,9 13,8 21,6 16,2 13,0
8,0 2.1
7,2 0 0.0
0,6- 0,7 6,9 5,4 2,9 4,2 2.1
0,7- 0,8 0,0 0,0 4,3 0,0 0.0
0,8- 0,9 3,4 0,0 1,4 1,8 0.0
0,9- 1,0 3,4 2,7 1,4 0,0 0.0
>1 0,0 0,0 0,0 0,0 0.0 Kit qud phdn tich hogt dp monooxygenase » mudi phdng thi nghifm.
Hai chdng mudi An. minimus phdng thi nghifm dupe tiln hdnh thd nhay cam vdi ca hai hod chit trong ndm 2009 vd ndm 2010 duoc xdc dinh hogt dp monooxygengse tgi cdc thdi diim tucmg dng. Kit qua trinh bdy teong bdng 7.
Hoat tinh monooxygenase d cd hai chung muoi null tgi phdng thi nghifm diu cho gia tri hogt
TAP CHi P H 6 N G C H O N G BgNH S 6 T RgT VA CAC BgNH Kf SINH T R O N G S 6 1-2012
dp monooxygenase nhd hon ngudng khdng (bdng 7). So sdnh hogt dd monooxygenase cda hai chdng theo tdng ndm, chung tdi nhdn thiy cd sy tdng hogt d§ cda enzyme ndy d chdng Hdg Son so vdi chung Tdn Lgc trong ndm 2009, 2010 (p<0,05)
Bdng 7. Kit qud phdn tich hogt df monooxygenase d cdc chung An.minimus phdng thi nghifm.
Quin Thi Quylt Chiln 2009 Hod Son 2009 Quyit Chien 2010 Hod son 2010
Hogt df trung binh (nmol/mg protein)
0,43 0,6 i t 0,49 0,54t
Dp tin cgy (95%)
0,09 0,11 0,065 0,071
Khodng bifn thien G l T B v d i d d t m c f y 9 5 %
0,34-0,52 0,50-0,72
0,43-0,56 a 0,47-0,61
Tuong tv nhu cdc quin thi muli tgi th\rc dia vd thi hf Fl cda chung, gid tri boat dp monooxygenase d hai chdng An. minimus phdng thi nghifm cOng dupe phdn tich theo cdc khodng gid tn hogt dp vd cd hai chdng diu cd ty If nhit dinh cdc cd thi phdn tich vupt ngudng khdng (bdng 8) nhu sau:
- Chung An. minimus Quylt Chiln-2009: 3,3%) - Chung An minimus Hod Son 2009:16,7%
- Chung ^n. minimus Quylt Chiln-2010: 4,3%) -Chdng ^n. mjHimwf Hod Son 2010: 14,3%
Bdng 8. Ty If so ca the quin thf (chiing PTN) phan theo cdc khodng gid ui hogt dp monooxygenase
Q u a n t h e \ ^ Tdn lac-2009 Hod Son-2009 Tan lgc-2010 Hod Son-2010
Gid tri hogt df monooxygenase theo cdc khoang (nmol/mg protein)
<0,I 0,0 1,0 0,0 4,8
0,1- 0,2 16,7 10,4 6,4 14,3
0,2- 0,3 16,7
9,4 19,1 7,1
0,3- 0,4 10,0
8,3 12,8 9,5
0,4- 0,5 30,0
6,3 21,3 11,9
0,5- 0,6 6,7 17,7 19,1 16,7
0,6- 0,7 3,3 17,7 6,4 4,8
0,7- 0,8 10,0
3,1 2,1 7,1
0,8- 0,9 3,3 9,4 8,5 9,5
0,9- 1,0 3,3 6,3 4,3 4,8
>1 0,0 10,4 0,0 9,5 3.2.3. Esterase
Kit qud phin Uch hogt Og esterase ciia cic quin thi tgi th\fc dja linh Hod Sinh.
Kit qud dupe trinh bdy d bdng 9, ngodi quin thi Anminimus tgi Bd Rd hogt dp esterase trung binh cua cdc quin thi diu <0,0738pmol/phut/mg protein. Nhu v$y theo ngudng ndy, cdc quin thS ndy nhgy cdm vdi vdi cdc hod chit dift cdn trdng thudc nhdm pyrethroid, nhdm phot pho hihi co, carbamate. Kit qud thd ndy khd tucmg ding vdi ket qud thd sinh hpc tai thvc dia, hiu hit cdc quin thi diu nhgy cdm vdi hai hod chit Id lambdacyhalothrin vd alphacypermethrin (vdi t^' If chit tuong dng trong thd sinh hpc 96- 100%).
Ket qua so sanh tuong quan thogt dd esterase cda cac quan the cdn lai so vdi quan the muSi tai Rdy cflng cho thiy cd sv tdng hogt tfaih esterase cda cdc quin thi ndy so vdi quin thf dRdy.
SO 1-2012 TAP CHi PH6NG CHONG BgNH S6l RgT VA CAC BgNH KY SINH TRONG 93
Bdng 9. Ket qud phdn tich hoat tinh esterase d cac quan the An. minimus thvc dia Hod Binh
Quan Thi Khdi, PC, TL B4Ra,NS, KB TnmgHmig, QC, TL Ray, VS, LS Chum, MT, LS
Hogt df trung binh (pmol/ph6t/mg
protein) 0,064 0,086 0,057 0,043 0,059
Dp tin c^y (95%) 0,006 0,014 0,011 0,007 0,012
Khoang biln thiin GTTB vdi CI 95%
0,058-0,07 0,072-0,10 0,046-0,068 0,036-0,050 0,047-0,071
Gid tri p so vdi quan the Ray,
VS,LS 0,003St 0,0004t 0,0134t 0,00731 Kit qua bdng 10 cho biit d6i chilu v6i ngudng khdng 0,073 8|amol/phut/mg protein trir quan thi t^ Ray thi cdc quin thi c6n lai diu c6 ty 1? nhat djnh so cd thi vupt nguSng khang. Quan th6 c6 ty 1$ % so cd the \ugt nguong khdng cao nhat la quan the tai Ba Rd, ddy cung la quin the c6 gid trj ho^t dp trung binh cao nhat trong cdc qudn the nghiSn curu (38,9%). Cu the ty 1^ % s6 cd thi vugt nguong khdng ciia cdc quan thi nhu sau
- Khcri, xa Phu Cudng, huyen Tan Lac : 4,2 % - Ba Rd, xd Ndt San, huy6n Kim Boi: 38,9%
- Trung Hung, xa Quylt Chien, huy$n Tan Lac: 5,9%
- Chum, xa My Thanh, huyen Lac Son: 6,3%
Bdng 10: Ty I? % s6 ca the quan thi phdn theo cac khoang gia trj hoat dp esterase
^ s ^ y If cd the cua
\ . timgQT Dia diim \^,^^
Khdi, PC, TL BaRd,NS,KB TningHimg, QC, TL Rdy, VS, LS Chum, MT, LS
Gid trj hogt dp esterase theo cac khoang (pmol/phut/mg protem)
<0,0I
3,19 0,00 0,00 4,26 0,00
0,01- 0,02 6,38 4,71 4,11 12,77 2,13
0,02- 0,03 8,51 1,18 10,96 27,66 31,91
0,03- 0,04 23,40
5,88 20,55 19,15 46,81
0,04- 0,05 12,77
0,05- 0,06 34,04 10,59| 15,29 31,51 34,04 8,51
10,96 2,13 0,00
0,06- 0,073
8,51 22,35 15,07 0,00 4,26
0,073- 0,08
2,13 25,88 2,74 0,00 4,26
0,08- 0,09 1,06 4,71 2,74 0,00 2,13
0,09- 0,1
0,00 4,71 0,00 0,00 0,00
>0,1
0,00 3,53 0,00 0,00 0,00 Kit qua phan tick hogt dp esterase & thi he Fl cia cdc quin thi
Chilu theo ngudng khdng 0,0738pmol/phut/mg protein, cd 1 quin thf FI cda Bd Rd cd gid tti hoat dp esterase ttung binh cao hon so vdi nguong khdng (bdng 10). Day cung Id quin the CO ty If muoi Fl chit thi;) nhit frong thd sjnh hpc vd quin thi tgi thon Rdy oho ket qud boat dp esterase trung binh thdp nhit cung Id quin thS cho ket qud thd nhay cam d thS hf FI Id nhgy vdi cd hai hod chit thd nghifm. Tuong tu enzyme monooxygenase, hogt dd enzyme esterase cung cd xu hudng tdng cao d cdc quan thi cd ty If chit thap hon frong thu sinh hpc.
Xft gid tri hogt dp esterase theo cdc khoang khdc nhau, chung tdi thu dupe t^ If sd ca thi d thf hf FI cda cdc quin thi vupt ngudng khdng nhu sau (bdng II): Quin thi Khdi: 7,6
%, Ba Ra: 24,3%, Trung Hung: 15,9%, Chum: 4,3% va Rdy: 0%
94 TAP CHl P H 6 N G C H 6 N G BgNH S 6 T RgT VACAC BgNH K t SINH T R O N G S O 1-2012
Bdng 11. Phdn tich hogt dp esterase d thf hfpl cda cac quin the/<«. mm/mMi thvc dja Hod Binh
Quin The KhdiFl,PC,TL B d R d F l , N S , K B Trung Hung F1,QC,TL RayFl,VS, LS Chum, MT, LS
Hogt dp trung binh (pmol/phul/mg Pr
0,072 0,078 0,064 0,038 0,065
Dp tin c§y (95%) 0,009 0,013 0,005 0,004 0,011
Khodng bien thiSn GTTB vdi Cl 95%
0,063-0,081 0,065-0,091 0,059-0,069 0,034-0,042 0,054-0,076
Gid ttj p so vdi quan thi Rdy,
VS,LS 0,0045t 0,0002t 0,004 iT 0,031 I t Bdng 12: Ty If s
Quan the ^~--^,^
KhdiFl,PC,TL B d R a F l , N S , K B Tning Hung F1,QC,TL RdyFl,VS, LS Chum, MT, LS
1 cd the quan Ihl (Fl) phdn theo cdc khodng gid tn hogt dp esterase Gid tri hogt df esterase theo cdc khodng ((imol/phut/mg protem)
<0,0110,01- 0,02 0,01 7,7 0,0 2,9 6,4 0,0
8,1 10,1 14,9 0,0
0,02- 0,03 5,1 5,4 2,9 21,3 8,5
0,03- 0,04 2,6 8,1 5,8 40.4 17,0
0,04- 0,05 25,6 10,8 20,3 4,3 12,8
0,05- 0,06
23,1 21,6 33,3 10,6 25,5
0,06- 0,073
28,2 21,6 8,7 2, 31,9
0,073- 0,08 J
5,1 16,2 14,5 0,0 4,3
0,08- 0,09 0,0 0,0 0,0 0,0 0,0
0,09- 0,1
2,6 0,0 0,0 0,0 0,0
>0,1 0,0 8,1 1,4 0,0 0,0 Ket qua phan tfch hoait dp esterase. 6 muSi ph6ng thf nghiem
Bdng 13: Ket qud phan ti'ch ho^t dp esterase d chung An. minimus phdng thi nghiem Quan The
Quylt Chiln-2009 Hod Son-2009 Quyet Chien-2010 Hoa Son-2010
Hogt df iTung binh (pmol/phut/mg protem)
D,(!68 0,072 0,071 0,082
DO tin cgy (95%) 0,005 0,007 0,009 0,013
Khodng bienthiSn GTTB vdi CI 95%
0,063-0,073 0,065-0,079 0,062-0,080 0,069-0,095 Theo kSt qua bdng 13, chiSu vdi ngudng khdng 0,0738nmol/phdt/mg protein, ehimg An.
minimus Hod Son cd gid tri hogt dp esterase tnmg binh cao hon ngudng khang trong ndm 2010 (0,082nmol/phut/mg protein). Chung An. minimus Quylt Chiln cho kit qud cd khd ndng khdng ttong thd sinh hpc vdi hai hod chit ttong ndm 2010 nhung khdng nhSn thiy sv tdng hogt tinh esterase
Bdng 14: Ty If so cd the quan th
Quan the ^ \ Tan lgc-2009 Hod Son-2009 Tan lgc-2010 Hod Son-2010
(Ichung PTN) phan theo cac khoang gid tri hogt dp Gid fri hogt dp EST theo cdc khoang (pmol/phdt/mg protem)
<0,01 0,0 1,1 2,1 0,0
0,01- 0,02 3,3 2,1 4,3 4,3
0,02- 0,03 0,0 2,1 0,0
0,0
0,03- 0,04
3,3 7,4 6,4
wl
0,04- 0,05 6,7 12,6 8,5 ,.,§•5
0,05- 0,06
16,7 21,1 17,0 21,3
0,06- 0,073
36,7 16,8 29,8 6,4
0,073- 0,08 26,7 31,6 25,5 25,5
0,08- 0,09 6,7 1,1 6,4 12,8
0,09- 0,1 0,0 1,1 0,0 2,1
>0,1 0,0 3,2 0,0
• 4.1 Ki Theo do ty If % so cdc thi vupt ngudng khdng ttong hai ndm nhu sau:
S6 1-2012 TAP CHi PHONG C H 6 N G BgNH S 6 T RET VA CAC BgNH KY SINH TRUNG
- Chung An. minimus Quylt Chiln-2009: 33,4%
- Chung .4n. minimus Hod Son 2009: 37,0%
- Chdng An. minimus Quylt Chiln-2010: 31,9%
- Chdng ^K, minimus Hod Son 2010: 44,7%
TAI L i f u THAM KHAO
1. NguySn Thi Be (2009)., Xac dinh muc nhgy cdm cda vie to sot ret vdi mpt so hod chat diet cdn frdng tgi cdc diim nghiSn cuu c6 djnh (sentilen) thupc ba miln d Vift Nam, Bdo cdo kit qua nghien cuu de tai cip Bp.
2. Vu Die Chinh (2005), Vec to sot r a d Vift Nam, phdn b l va dp nhgy cdm ciia chung vdi mft so hod chat diet cdn triing frong phdng chong sit ret, Lugn vin thgc sT cao hoc 2005.
3. Josiane Etang, Lucien Manga, Jean-Claude Toto, Pieue Guillet, Etiemie Fondjo and Fabrice Chandre, (2007), "Specfrum of metabolic-based resistance to DDT and pyrethroids m Anopheles gambiae s.l. populations fro Cameroon", Journal of Vector Ecology, Vol. 32 no
l,pp. 123-134.
4. Liverpool School of Tropical Medicine, (2002), Technical file.
5. WHO, (1998), Techniques lo deled insecticide resistance mechanism, WHO/CDS/CPC/MAL/98.6.
6. WHO, (1998), Test procedure for insecticide resistance monitoring in malaria vectors, bio- efficacy and persistence of insecticide on treated surfaces, WHO/CDS/CPC/MAL/98.12
E V A L U A T I O N O F SUSCEPTIBILITY OF ANOPHELES MINIMUS FROM I N S C E T A T I U M AND ANOPHELES MINIMUS POPULATIONS IN H O A BINH P R O V I N C E T O I N S E C T I C I D E BY W H O TEST AND B I O C H E M I C A L ASSAY
Dao Minh Trang, Ho Dinh Trung National Institute of Malariology Parasitology Entomology Abstracts
This study was carried out in 2009 -2011 with 5 Anopheles minimus populations from Hoa Binh and 2 An. minimus strains in insectarium. The bioassay results showed that 4 field populations were susceptible to Lambdacyhalothrin and Alphacypermethrin (98-100%
mortality), 1 field populations were susceptible lo Alphacypermethrin, possible resistance to Lambdacyhalothrin (96% mortality). Mostly, mortalities in bioassays in FJ generations with these insecticide is lower (66-10% mortality). An. minimus Hoa Son strain was resistance to Lambdacyhalothrin and Alphacypermethrin in 2 times (2009, 2010). An. minimus Quyet Chien strain was susceptible to Lambdacyhalothrin and Alphacypermethrin in 2009 and possible resistance to Lambdacyhalothrin and Alphacypermethrin in 2010. The levels of esterase and monooxygenase had tendency to increase in populations having lower molalities in bioassays at filial generation (Fl)
C&a bO phdn bi§n TS. Phgm Thj Khoa Vifn Sot rit-KST-CT TlT
