•J TAPCHIKHOAHOCOHSPTPHCM Nguyin Thi TJtutmg Huyin va tgk
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BANH GIA ANH HirdNG CUA KEM LEN SU" S 6 N G CA NGUA VAN GIAI BOAN AU TRUNG (/ - 7i,gay /u5«)
NGUYEN THj THl/CTNG HUYgN', B O A N LE MINH HI^N"
TOM TAT
.^^^''^f^'-g^ip ciu nay nhdm ddnh gid inh huo,^ cia ning dd kem Ien susA„„
^au trungcaN v^O-7ngay tuit) Phdi edNgZlmdi Ihudl * ^ X , S d ^ m mua, kem alOnong do khac nhau (I-IO mgfL) vi 16 dii chung (0 m^L^rZTdi
'•^'^^'^{^•rmh ttin dodn tile sing eua du trimg ci Ngua vinlheoMZ^
luang tac cua nong do Zn vi ihdi gian mai xde dinh duac gid tri LcZZ^hts^duZ^
m ^ ^ ^ ^ l a n g cua nong 0Zn khdo sdt va thai gian nuoi ^ K uyen f ^ F ' * * * * ' • • ' * ' ' « " ' ^ ^ ° " « ™ ' ' « ° ^ - ^ P <im, i u ttung c i Ngua van, kim loai nang
ABSTRACT
Evaluating the mfluences of zinc (Zn) on the life gf larval zebrafish (1-7 days old) afzehZlT, '^"""/'"fy « ' " " " ' " " ' ^ '*" i'lfluences of concenuations ofZn on the life L ^ ^ f "-'' "^^ " ' * '*^" "^""^ " " ^ ^ " - - ^ ^ e 1 t o l l in Ihe
a t fe^TZTT'^'r'''"''"''''"''''•^^^'''"''^^''-^-•"'dconeen^^^^^
ofZn. respecttvely The resulls show: (i) the minimal concentration of Zn affected Z
^""I ate gfzebrgfish larvae fallowing interaction between concentrations of ^uTd cuhure time can calculate LCtso gfZn to the larval zebrafish stage (1-7 dZ-immth
Keywords: zebrafish, larval zebrafish, zinc pollution, heartbeat, heavy metal.
!• Gioi thifu
nh„ . ? "h^f" " ^ !'«*"8 e«y nhiiu inh hudng nguy hai eho sue khde eon ngudi cung nhu cac smh vat khac, ttong dd d nhiem kim loai nang d mdi ttudng nude g i ^ h a i 2k.
ma tteu bieu la o nh,dm cadmium, asen, ehi, kem... Die bipt, k i m S i i n a n g ^ S S nhap va ttch tp ttong ea flie ngudi v i sinh vjt mdt each ttuc tiip' hay gZ T ^ Z
rryl^h^if^ttr ^ "^ '-''" " '""' °- ^^J.'^"^Z
-^IJUfV^.^ T ^'' '*^'" ™ ' = " ' ^ * huyenntth@hcmup edu vn HVCKTRrtngOaihoc Khoa hoc TunhiSn. BHQG TPHCM
103
TAP CHl KHOA HOC BHSP TPHCM SS 6(84) nam 2016
Trong cic kim loai, kem la mpt nguyen td vi luang thiit yiu cho ddi sdng cua ca thuc vat, ddng vjt vi con ngudi. Zn dupe tim thiy trong khip cae ti bio cua ca thi kirn loai niy tham gia vio eiu tao cua hon 200 loai enzyme frong ca thi ngudi, gdp phan dieu hda chuyen hda lipid, tham gia tao miu cung vdi sit, cin thiit cho su biel hda te bio, giflp dn dinh cau tnic mang ti bao, tham gia vao ehfle ning eua he miln djch. .^ Thieu Zn se giy ra eic ttieu chung nhu: chan an, chjm ting trudng, suy giam he miin djch, tiiiiu ning sinh due, rung toe va long, suy giam vj giic va khuu giic...
Tuy nhien, khi co the thfla Zn hojc ndng dp Zn eao frong mdi frudng sing eung gay hai cho cic sinh vjt d nhung noi bj d nhiem [10]. Do dd, viec nghien cflu mflc dp anh hfldng eua Zn len su sinh ttudng vi phat triin ciia ca flii sinh vat la vd cung cip tiiiit.
Tfl nhung nam 1970, ei Ngua vSn da dupe nha khoa hpe George Streisinger dua vio phuc vp cho nghien cflu khoa hpe. Diy la ddi tupng dupe sfl dung nhiiu ttong cac till nghiem vi ehung cd nhflng uu diim nhu: kich thudc nhd, khi nang sinh sin ldn, thu tinh ngoii, phdi ldn vi ttong suit, phat ttiin nhanh, de nudi [9, 10]. Trong cic giai do?n phit ttien ciia ca Ngua vin, giai doan phdi vi iu trflng cd thdi gian phat ttiin nhanh nhit vi dp nhay cim cao nhit. Chi vdi mdt ndng dp nhd hda ehit tic dpng len giai doan niy, ta ciing de dang nhjn thiy duac su thay ddi d chflng. Vi tiii, phdi va iu trung li rapt giai doan kha dl dupc lua chpn di fliue hien nghien cflu. Nghien cflu nay su dpng phdi ea Ngua vin de danh gii inh hudng cua Zn len su sdng efla giai doan lu trung (1-7 ngiy tudi).
2. Vjt li^u va phvffng phap nghien cftu 2.1. Hoa chil
, „ . ^f\fj?,^' ^^ " ^ ' ^ ^^ •=*• ^"^'^ (^'8™^): hda ehit di ehuin pH: NaOH IN (Merck), HCI IN (Merck).
7„ri °r^ ''lon T ' ' ^" ^""^ P*"" '^ "™^ •*' ' ^ hing each hda tan 0,2086g muii IflZZ^ 'OOmL nuac cat. Dung djch sau khi pha dupe dung ttong chai DuraS s,eh, Sudpngstoeknaydepharacaendngddfliinghiera(I,2,3,4,5,6,7 8 9 lOmgfl!) s,ch e ' l t d c t i t t n V ' " ' " " " ' ' *'• "''•'*•" •' ™*^ "^^ '"'' '' '"^^ '"-'' ^ ' 2.2. Vit lif u
2.3. ^ w 4 X ' ° ' " ""' ""''"" ' ' " * ' ""' • ' ''"^- P""' ''• ^' "P*^-
vit K l ^ f ' i l ' t f T * ^ n ^ . ' " ""'""^ ™ "«"''"' °'*" P * - Sh* U Ngudi vi Ddng d l ^ ' k l t i ™ t ^ ^ T ! " ' ' ' " P ' " " ™ ^ ^ - '^^ '^W^ '^ hd me dupc nudiin d,nh dieu kipn song tfieo ehu k, quang 14 gid sang:IO gid tdi tai phdng ti,i nghiem.
Phuang phdp phoi cava Ihu phdi
ehu k i ' ^ i ' Z f e h o t " ? ? ' " ' ^ '"^*' ''^ '^'" r^ng '^i <JVC vi ci cai flieo ti le 1: 2. T,o toll bd PM duoc tii?" " " ' T " ' '*'"'™ '^ "h^" t"J- S - 3 - 5 phfli cho phii, toan bp phoi dupc tiiu sang coc ttiuy tinh lOOOmL vi tiin hinh phin lopi phdi. Cae 104
««
TAP CHi KHOA HOC BHSP TPHCM
Nguyin Thi Thucmg Huyin vi Igk
phdi tdt^dflgc thu nhan vao cae dTa petti thfly tinh sach cd dfldng kinh 35mm d i nudi - Phuang phap gdy nhiim Zn cho iu triing
. .Ll^.oh^_ cae phdi tdt phuc vu cho flii nghiem Tit c i cie phdi flu nghiem diu duoc nuoi o nhipt dd 28 "C, pH = 7 5 CWesterfiplH inn-n oi.- A ' " ,"K™'™ ° ^ " "upc
„ f V • • ' , , .^"f" ''^'•'^c^^™oia, 2007). Pho, duoc tiep xuc VOT mdi midnB chua Zn o cac 10 ndng dd (1 2 3 4 5 fi 7 a o „« ,n ,„ /r > • , - i"- , . ^
^ d p t o ^ bp dfa niy vao ede tfiuy tinh chua 300ml mdi ttu4 vdi eic n d n T d o l n ttrong ung de khao sat ti le sdng, nhjp tim, ehiiu dii tfl ngay tiifl I - din ngay fl,fl 7
tJanh gia sue song cua du trimg
Thdng ke ti le sdng chit v i ghi nhjn eic di tat cua i u frung d tflng ndna dd khoang each g,ua cic lin ghi nhan sau mdi 24 gid (iu trflng tfl ngiy f d i n ^ l y 7 " ' ' '
^ _ 1 Phvangphip dim nhip tim
^ A u ttung dupc hflt bang dng hut nhua mim vdi dudng kinh I mm Sau do chn vao iia petti dudng kinh 60mm cd ehfla du mdi trudng ttro^tg d i T u S ^ g cd thi t t g s i T u t r ' " ' " " '".""• ' " " ^ ^ ' ™ « ' P " ' - '^hPn ngiu nhien 6 i u tti^long tong so au tnmg song qua tmig ngiy, tu ngiy 1 din ngay 7. Thi nghiem lap lai 5 fin
Phirong phdp do kich thuac au triing
Au trung dupc do kieh thude qua tung ngay (tu neiv 1 din m^av 7, r^r, nhten 6 i u ttung ttong tdng sd iu tttag sdng'thf nghie^mfi'l^i / L ^ ^ ' '^- ' ' ' " " " « ^ "
petti d l r L ^ f n h ' ^ , ' ' " ' "^l ^ " ! ' ' " " " ^ " ' ^ " - " " ^ S '<'•'* imm. Sau dd cho vao dTa dup D r m l ^ i r k l ' h ' ' ' ^ " ' - ' " ^ ' ^ •"""« ™« "* ^ " ^ e ^°"8 CO thi sdng chup h f r i a T " " ' " " ' " ^^ "^ " * i "'"h' • « ' " hd giae XI vf
Hinh L Chieu dai ca thi du trimg ca Ngua vdn (X2)
TAP CHi KHOA HOC BHSP TPHCM SS 6(84) nim 2016
Cieh do ehieu dai; chieu dii eua iu tning dupe tinh hi phin mieng din phin dudi (k6t thue xflong, co). Vay dudi khdng duac titJi vio kich thude iu tning.
Phuong phap quy ddi kieh tiiude iu ttxmg; ta'eh tiiudc cua iu trflng sau khi dupc do bSng thudc do cfla kinh hiin vi se dupe quy ddi sang don vi mm bing cdng fliiic sau
1 A L = — X -
10 fl trong dd;
- L; chieu dii co the ci (mm);
- A: so v^ch do dupc tu thudc do cua kinh hiin vi soi ndi;
- B; bdi giic vjt kinh.
- Phtrang phdp xuU thing kl
Tit ci sd Ueu cfla de tii dupe xu Ii flieo cae fliujt toin xic suit flidng ke tten may VI tmh bang phan mem Minitab 16, SPSS 20. Cic si liju ttung binh dupc ttinh biy 6 d?ng X ± SE. Xfl U sai khac vi ti lp sdng chit iu tiling, nhjp tim vi chiiu dai iu ttiing bing phuong phip phin tieh phuang sai mpt nhan td; ANOVA - One Way.
De phan tich mii lien hj gifta ti le sdng cua iu frung vdi eic yiu t6 ning dp Zn vi fliOT gian, chflng tdi ap dpng phuang phip phin tich hdi quy logistic (logistic regression analysis metiiod) vdi md hinh hdi quy dupc sfl dpng li Poission regression model dupe phit biiu dudi dang: log (-^ ] = a -i- ySx,
Khi dd thdng si (parameter) a vi p ttong md hinh dupe udc tmh (estimation) flieo phuong phip hpp I, cpc d^i (maximum likelihood-based mefl,od).
fel fcl
.'•=1 .-=1
3. Kit qua vi thao lujn
3.L Anh hudng cua Zn lin lil^ sing cua iu trimg
li iu SnTttoiri t°^t" '^^ ' * " ° ^ " ^ ^ "^ '^^° "hit ttong vdng ddi ei, djc bijt n ^ c t a v^ Vn „ ' ''K-'" t ' ? ' *"> ^^' "} ^ ' "hjn sau m§i 24 gid di danh gii dj
::i^'di:::irdu;sfb™d Bt^t"'^ ^'"« ^'^'^'" ^
TAP CHl KHOA HOC BHSP TPHCM Nguyin Thi Thuang Huyin vi Igk
Bang I. Till song cua du trung ci Ngua vin qua cdc mic thdt gian a cdc nong do khio sat
70,00 66,00 58,67 54,00 44,67 43,33 37,33 29,33 26,67 2,52
(134) 8S,67 2
2,59 (133) H 88.00
^ P (132)
" ^ 84,00
± 2,99 (126) 81,33
± 3,18 (122) 77,33
3,42 (116) 65,33
3,89 (98)
3,52 (113) 74,67
3,55 (112) 74,67
± 3,55 (112) 74,67
± 3,55 (112) 74,67
± 3,55 (112) 66,00
± 3.87 (99) 54,67
± 4,06 (82)
3,74 (105) 69,33
± 3,76 (104) 69,33 3,76 (104) 68,00
± 3,81 (102) 67,33
± 3,83 (101) 62,00
± 3,96 (93) 52,00 4,08 (78)
3,87 (99) 63,33
± 3,93 (95) 60,00
± 4,00 (90) 60,00
± 4,00 (90) 57,33
± 4,04 (86) 50,67 4,08 (76) 34,67
± 3,89 (52)
4,02 (88) 55,33
± 4,06 (83) 52,00
± 4,08 (78) 47,33 4,08 (71) 44,67
± 4,06 (67) 39,33
± 3.99 (59) 28,67
± 3,69 (43)
4,07 (81) 47,33
± 4,08 (71) 42,00
± 4,03 (63) 39,33
± 3,99 (59) 34,67
± 3,89 (52) 27,33
± 3,64 (41) 20,67
± 3,31 (31)
4,06 (67) 30,00
± 3,74 (45) 27,33
± 3,64 (41) 24,67
± 3,52 (37) 16,67
± 3,04 (25) 10,67
± 2,52 (16) 4,00
± 1,60 (6)
4,05 (65) 24,00
± 3,49 (36) 20,67
± 3,31 (31) 15,33 2,94 (23) 12,00
± 2,65 (18) 8,00
2,22 (12) 3,33 1,47 (5)
3,95 (56) 15,33
± 2,94 (23) 10,00
± 2,45 (15) 4,67
1,72 (7) 4,00
± 1,60 (6) 0,67
0,66 (1) 0,00
± 0,00 (0)
3.72
8,67
2,30
4,67
± 1,72
1,33
0,94
0,67
± 0,66
(1) 0,00
± 0,00
(0) 0,00 0,00 (0)
3,61
8,67
2,30
0,00
0,00
0,00
0,00
0,00
0,00
0.00
0,00
0,00
± 0,00
(0)
Qua Bang 1, ta nhjn fliiy ti Ip sdng cfla iu ttflng ci Ngua vin giam tuyin tinh tiieo su tang din cua ndng dO Zn frong mdi ttudng qua cac ngay nudi, su giim nay cd « nghia ve mjt flidng ke vdi dp tin cjy eao (p = 0,00). Tfl kit qua niy, tiin hanh danh gia cic yeu to flidi gian nudi vi ndng dp Zn len ti le sdng cfla iu tiling
TAP CHl KHOA HOC BHSP TPHCM SS 6(84) nam 2016
3.1.1. Khao sdt su biin dgng ti Ii sing cua du trimg qua cdc ngdy nuoi trong mdi tnrdng cd Zn d cic ndng do khao sdt
Tfl ket qui Bang I, ehflng tdi tiin hinh danh gia su biin ddng ti le s i n g cua iu tiling c i Ngua vin qua cac ngiy nudi ttong cic mdi ttudng cd chfla eac ndng dd Zn khio sit. Ket q u i t h i hien d Bang 2.
Bing 2. Anh hudng cua thai gian nudi din ti li sing cua iu trimg cd a cac ndng do Zn Ndng d9
Zn (mB«.)
BC
1
4
8
9
v l u \h tien doin (predictor) H^Dg so a N s i y - B H^ng s d a
N g 4 y - p H^ng so a N g i y - P HSng sd a N g 4 y - P H9Dg sd a
N g i v - B HSng sd a
H2ngsda N E S V - P M^ng sd a
N E 4 V - P H9ngsda Ngiv - p Hing s d a Ng4v-B tt^ng so a N B J V - P
He sS iroc l u ^ g -0,026 -0,044 -0,190 -0,040 -0,273 -0,039 -0,279 -0,078 -0,384 -0,100 -0,445 -0,141 -0,493 -0,290 -0,549 -0,348 -0,331 -0,688 -0,221 -1,027 0,365 -1,624
P 0,729 0,010 0,017 0,029 0,001 0,043 0,001 0,000 0,000 0,000 0,000 0,000 0,000 0.000 0.000 0,000 0,060 0,000 0,360 0,000 0,282 0,000
Risk Ratio
0,957
0,961
0.962
0,925
0,905
0,868
0,749
0,706
0,503
0,338
0,197
95% Cl
0,925 - 0,989
0,927 - 0,996
0,927 - 0,999
0,888 - 0,963
0,865 - 0,946
0,826-0,913
0,698 - 0,803
0,651 -0,766
0.434 - 0,583
0,275 - 0,465
0,121-0,321
i u ttt^g c f f ; ; i l i e ' b i * t t a ^ ™ * " ^ ' " ™ ^ ' * ™ « '° 2 " ('* « ' ^ h ™ g ) . « i j sdng eua n n, I Z ^ ^ l „ ?'^ . h " ' 1 " " "^^Wt nudi, su khae biet n i v ed v n„hr„ ,hA„„ J i „ = sp khae bipt n i y ed y nghia tfidng ke (p =
108
- ! I ! ^ ' 1 ^ ^ * ^ ^ ^ ^ ' ^ ^ ^ ^ ' S P ™ C M ^ ^ ; ^ ^
^ ' ^ 8 giai dtjan iu ttung, ca bi anh hudng ttuc flip bdi mdi ttudng xung ouanh (inh
^* ^ g Z l 2 7 ^ £ i T 'tit^.':'^'^ -^ " ^ n i i n g ' S r b i o l ; ' ' n f £ uung song Chu yeu dpa vao ehat dmh duong tir ttii noSn hoang cho din khi liv Li^e
* S l t \ 7 ? - ^"^ " h i ^ " ^ - ^ o ^ 5 - 6 ngay tudi, ttii'noL h « 1 tid^
. ciNgpava„tiieo%^*a"it^^:Sq-:^^r.trg'r"«^"^'^«
tai cdc moc thai gian
Thiri gian
Ngiy I
Ngiy 2
Ngiy 3
Ngiy 4
Ngiy 5
Ngiy 6
Ngiy 7
Vea to (jen tfoin (prediclor) Hing so a Ndng d $ - B
Iting so o Nong d$ - p t l i n g so a Ndng dp - B Hing sd o Ndng d$ - B Hing sd a Ndng d$ - B Hing s d a N d n g d 9 - B H i n g s d a Ndng d$ - B
H f s d u ^ Iimng
•0,113 -0,113 0,014
• 0 3 8 0,059 -0J50 0,065 -0,278 0.060 -OJOI 4,706E^*"
•0J36 -0.173 -0J77
P 0,037 0,000 0.802 0.000 0.289 0,000 0 J 4 7 0,000 0 J 9 6 0,000 0,999 0,000 0,011 0,000
RiskRnho
0,893
0,812
0,779
0,757
0,740
0,715
0,686
95% Cl
0,874-0,913
0,791 - 0,834
0,757 - 0,801
0,735 - 0,781
0,716-0,764
0,689-0,741
0,656-0,717
TAP CHi KHOA HOC DHSP TPHCM Sd 6(84) nam 2016
Ngay tii ngay thii 1, ti le song cua au trung tii nong do 6 - 1 0 mg/L deu nam duoi ngudng 50% ( B ^ g 1), khac biet co y nghTa th6ng ke (p = 0,00). Cu th§, cii tang Img/L Znthi ti 1$ s6ng ciia ku trung giam di e"**'' '•* ^ 0,893 liin. IGii xii li va phan tich kit qua 6 Bang 3 tEti cac ngay tir thii 2 den thii 7 cung cho k6t qua tucmg tir ngay thii 1. Vi vSy, c6 the ket luan siic song cua Su trung ca Ngira van vao ngay thii 1 sau khi no co miic dO nhgy cam cao nhat voi Zn. DiSu n^y co thl do khi con d giai doan ph6i, ca dugc bao v§ trong Idp mang dem nen hgn che dugc s\r xam nh|ip cua Zn ngoai moi tni6ng vao ben trong phoi. Tuy nhien, sau khi ca no, Zn trong moi tniong nu(M se tac d6ng true tiSp tr6n co the ca, gay chet khi cr nong do cang cao. Nghien ciiu cua Hwang va cs.
(1995) da khSng dinh mtic do nhay cam dfii vdi Zn cr giai doan iu tmng co thS co lien quan toi sir phat triln cua Su trung, d6ng thoi tinh thdm ciia mang t l bao d6i vdi cac ion hoac CO che cSn bang ion cung co thl bi thay d6i trong qua trinli phdt triln ciia iu trung. O giai dogn ku triing, co the khi co sir thay d6i ndng dO gay nhilm Zn se lam thay ddi miic dp nhay cam vol Zn [4, 5, 6].
Ngoii ra, moi trudng nuoi ca vdi cac ylu td sinh thai (anh sang, tilng 6n, ndng dp O2 va CO2 hoa tan) chua duoc dilu chinh tdi im cilng co thl la mot trong nhOng tac nhan gay stress cho ca. Kit qua cua chiing toi phii hgrp vdi cdng bd cua Hwang va cs.
(1995) [6], hQ cho rSng du triing co tinh nhay cam cao nhdt trong vong ddi ca, sy: nhay cim Clia du triing liic mdi nd tang len dudi sir bdt lgi cua mdi tnidng d nhilm kim lo?!
nhuZn.
3.1.3. Khdo sdt dnh huang cua hai yiu td thai gian vd ndng dg Zn den ti I? song cua du trimg
Til ket qua Bang 1, chiing tdi danh gid su biln ddng ti le sdng cua du triing cd Ngv:a van theo sir anh hudng tuong tac ciia hai ylu td thdi gian va ndng do Zn theo mo hinh hdi quy Poission (Poission regression) dugc phat bilu nhu sau:
Log(|ii/Ni) - (Thdi gian + Nong dO)
Ho§c: Log(ni/Ni) = a + pi *Ngay + p2* Nong dp + |33*Ngay* Ndng dp Ket qui the hi?n qua Bang 4.
Tiin dodn gid trf LCjo or ngdy 1
Bdng 4. Anh huang tuang tdc cua thdi gian vd ndng dQ Zn den ti 1$ sdng cua du trung cd Ng^a vdn Ylu td lien doin
(prediclor) Hing sd 0 (constanO
Ngiv-BI Ndngd$-p2 Tuvng lic ~ p3
H^sd ud^c lin)rng
-0,113 0,000 -0,113 0,000
P 0,037
0,000
Risk Ratio
0,893 1,000 0,893 1,000
95% Cl
»4
TAPCHi KHOA HOC BHSP TPHCM Nguyin Thi Thutmg Huyin vi tgk
.^•.n S . f f '""* °. ^ * ^ ''• "^'^^ •*' '^'*' '«> P ' ' " " ^ • * * ti™ i l " ^ « 1« Sdng Clia i u ttung (Pr) flieo su anh hudng cua cic yeu td nhu sau:
Pr = exp(a -I- pi 'Thdi gian -I- p2«Ndng dp + p3*Ndng dd'Thdi gian) ttinh ^ ^ ^ ° : " ; - " ' ^ ' ° "•"'"'= f*^ = •*'•' ' ^ ' P3 = «•<""'• ™ ^io <^ "J^™ phttong
ttinh tten doan n le sdng nhu sau: ' Pr = exp(-0,l 13 - 0,113* Ndng do)
.... P ^ " ? ° ° . ' ^ "^".t^ <^° * « tinh toin dugc ngudng ndng dd Zn giy chit 50%
^ 1 vo, au tnuig ca Ngya van (li ngudng ndng do Zn m i tai dd tt' le sdng c L i u t i g ca Ngua van glim di 50%) nhu sau: uo au iruiig
Ti \i sdng udc tinh tai ngay 1 ttong mdi ttudng khdng cd Zn li:
Pi<0) (Thdi gian = I, Zn = 0) = exp(-0,113) = 0,893 Ti le sdng udc tinh tai ngiy 1 frong mdi ttudng cd ion Zn Ii:
Pr(X) (Thdi gian = 1, Zn = X) = exp(-0,l 13 - 0 , n 3 * X )
Ti le sdng cua i u ttimg c i Ngua vin giim di 50% nghia l i Pi(X)/Pi<0) = •/, = 0 5 Nhu vay: Pr(X)/Pi(0) = 0,5 -^ exp(-0,113 - 0,113»Xy 0,893= 0,5
- exp(-0,l 13 - 0,113«X) = 0,447 - - -0,113 - 0,113'X = Ln(0,447)
— X = 6,12nig/LZn
ttiwiT* ' ^ ' • ^ •.*' " " * ' ° ^ ^"'^ "S^^i nSng "Jp Zn gay chSt 50% ddi vdi i u ttung ca Ngua van o ngiy 2 den ngiy 7. Kdt qui dugc thJ hipi d Bing 5
Bing 5. Gii tri LCio cua au trimg ca Ngua vdn qua timg ngay Igi cic ning dg Zn khao sdt
Thdi gian Nong do giy chet 50% au triing (LCs.)
I l l
TAP CHl KHOA HOC BHSP TPHCM SS 6(84) nim 2016
nudi dudi tic ddng cua Zn. Ddn ngiy thii 7, chi cin rncit lugng Zn Ii 1,77 mg/L ciing lim cho sd lugng iu ttiing chSt hon 50%. Theo QCVN 08-MT:2015/BTNMT, ham lugng Zn cho phep ttong nudc sinh hoat va ao hd cho ddng vat thiiy sinh dao ddng tir 0,5 - 1,0 mg/L [1]. Cic gii LCso tinh dugc qua thuc nghiSm ddu vugt nguong so vdi gidi han cho phep.
3.2. Nhjp fim cua iu tring ca Ngira vin tai cic thdi tiim vi cac ning dp Zn khdo sil
Dua vio ket qua vd ti le sdng cua iu ttiing d Bing 1 vi Bing 5, bit diu tit ndng dd 6 mg/L ngay tai ngay nudi thu nhit tt le sdng cua au ttiing da chit vugt nguong 50%, nen chung tdi chi khio sat nhip tim ttong giai d o ^ niy tai cac ndng dd 1 - 5 mg/L vi Id ddi chung qua mdi ngay (tir ngiy tfiu: 1 din ngiy thOr 7). Kit qui tfii hien d Bang 6.
Bing 6. Nhip tim cua au trimg ci Ngva vdn Igi cdc nghiem thiic khao sal
Ngiy
1 2
3 4 5
BC 277,00 ±
2,86"
272,20 ± 4,72"
241.80±
5.42"
232.60 ± 6,03*
206,60 ± 4,19"
188,20 >t 4,02"' 172,60 ±
2,40"
Nlijp lim Su triing or cic nOne d$ Zn Imp/I.) t
261,60 ±4,03'"
246,00
±553l»|3 234,20 ± 6 , 9 r * 213,00 ±
4 , 4 2 * 195,60 i 4,40**
178,80 ± 3,46**
169,00 ± 4 , 1 5 "
a. b. c. d: Ihihi4n a. A y. S. e: IhS hie
2 2 4 0 , 2 0 ± 3 , 7 I "
239,20 ± 5 , 9 3 "
210,80 ± 5,49'*
192,60
±3,88"*^
184,60 ± 5,04'"
172,20
±4,60'"'^
162,60 ± 3,42"'
3 235,00 ±
3,70"
234,00 ± 6,60""
204,20 ± 5,91*
191,40±
5 , 5 8 * 176.80 ±
5 , 7 4 "
160,40 ± 2,64"
156,20 ± 2,53"
sv khac bi?l Iheo hang & t?(3 tin cgy n stf khac bi^l Iheo col a d^ lin cpy
214,60 ± 4 , 6 2 "
208,20 ± 5,04"
202,80 ± 5 . 7 1 "
158,40 ± 156,40 ± 4,82*
134,80 ± I31,80±
2,95"
95%
95%
207,00 ± 2,74'"
202,20 ± 199,60 ± 8,86"
156,00 ± 153.20 ± 119,60 ± 98,40 ±
7 „ „ J . ? ^ - ' ° ? " * T ' " ' ^ « ^ ' ^ ' * * " ^ y ' ' * ' P ' ™ ' = * i ^ ^ " t r t n g o 4 N g u a vin ttong I i ^ m o ^ L f d ? T " ^ '^° ^ "^^ - ^ ^"^ "^"8 " ' Z" -^""S md!ttucmfnudi, ttoni cung mgt nong dg nhjp ttm au ttung COng giim din qua cac ngiy nudi. Cu thi tUt^^^tvl- ^'^l '•'" ' " " ^ " • * '="" ^" "^"S * '« <•« <=l'ung la 277 ± 2,86 S ^ f & L w 7 r " ^ . ' ' ' ^" •"' '™ " " "^ =»ini hon vi biin thien theo sv T l ^ S m T l ' ^ " . ^ ' r . ' " " ^ '" ' ^ 8 » t o «• 05,40 - 70 nhip/phut) t i ndng dp tu 1 5mg/L so vdi doi chimg, su khac bift niy cd y nghia vi mit fting kd (p <
169 00™4 15 nh / K ; ? ' ' "'^ *• "^P " " " ^ S • ' ' * ^' nSng dd 1 mg/L Ii J '^''•V" *''•'S nhip/phut va glim xudng tdi gii tti thin nhit li9S 40+ 9 1^ i,- / ..
•hdldin ngay t h d 7 U i t qui dugc S t n t ; £ r r " ' ' " ° ' " ' ' ^ " " ' ' ^ ' ' ' " « * ^ _ _ _ J « ^ ^ S ^ ^ ^ ^ * ™ ^ < ™ ^ ? c<i Afei,^ vi„ to/ cdc nghiim Ihuc khdo sdt
Ngiy Kicll lliln>c i n Irilng fr cic ning d6 Zn (mg/l. 1
• ^ J 2
o. ». c rf. 'belii!rsrkhacbiellheohdngai>,licay95%
°. <'• '•• 'fe' *W" W W c bill lh,„ cot &doli„cav 95%
TAP CHi KHOA HOC BHSP TPHCM Sd 6(84) nim 2016
Ket qui Bing 7 cho thay, chieu dai trung binh cua au triing giim theo tuyen linh theo su tang din ciia ndng do Zn qua cic ngiy. Cu the:
Tai ngiy thu nhit, chiiu dai cua iu trimg dugc giy nhiim Zn d cic ndng do 4 vi 5 mg/L ngan hgn vi su khic biet niy cd y nghia vd mat thdng ke (p < 0,05) so vdi chieu dai ciia cic iu trimg d Id ddi chiing. Chieu dai ttung binh d ndng do 1; 2 va 3 mg/L khic biet khdng cd y nghta vi mgt thing kd so vdi Id ddi chimg (p > 0,05). Bit diu tit ndng do 4 mg/L, chieu dii mdi cd su khac biet cd y nghTa ve mat thing k€ so vdi Id ddi chung (p = 0,02). Cu the, chieu dai cua iu trimg d mdi truang cd ndng do 4mg/L ngin hon Id ddi chung li 0,14 mm; vdi 95% dao ddng ttong khoing 0,01 - 0,26 mm (p = 0,028). Chiiu dai ttung binh fliip nhit d ndng dg 5 mg/L (3,48 ± 0,03 mm) va khic biet cd y nghia ve mat thong ke so vdi Id ddi chiing (p < 0,05).
Tinh den ngay thu 7, chieu dai d cic ndng do 2; 3; 4 va 5 mg/L cd khac biet c6 y nghta vi mat tfiong ke so vdi Id ddi chiing vdi dg tin cay cao (p = 0,00). Ndng dg Img/L cd khic bi|t so vdi Id ddi chimg nhung khdng cd ^ nghia vi mit thing ke (p >
0,05). So sinh chiiu dii iu ttimg giua ndng do 1 mg/L vi 2 mg/L cd sy khac biet cd y nghTa ve mat tfidng ke (p = 0,00), giita cic ndng do ki can cdn Iai cd khac biet nhung khdng cd ^ nghTa tfiong ke (p > 0,05). Trong cic ndng dg Zn khio sit, ndng dO 5 mg/L anh hudng m ^ nhit din su tfiay ddi chiiu dii iu triing ci Ngua vin. C\f thi, chiiu dii Clia iu ttung d mdi ttudng cd nong do Zn 5 mg/L dat gii ttj fliip nhit ttong tit ci cic ndng do (3,60 mm ± 0,02), ngin hon Id ddi chimg li 0,38 mm (p = 0,00).
Nhu viy, tt'nh tii ngiy thii nhit sau khi bit diu nudi, chung tdi nhin tfiiy sv ting din nong do Zn da inh hudng din kich tfiudc cua iu ttiing ca Ngua vin, Zn di Iam chim ting ttudng d au ttung ci Ngua vin. Cac ndng do I; 2 va 3 mg/L chua cd su inh hudng id tel den kich ttiudc ciia au ming. Cac ndng do 4 va 5 mg/L cd anh hudng manh nhit ttong cic ndng do khio sit. NguySn nhan cd thi la do co tfii ca cd xu hudng hip thu cic ion tir mdi trudng ben ngoai (qua ci hai con dudng: hip tfiu ttuc ttip hay qua dudng tfeu hda). MSc du chiing ludn cd co che de tv loai bd, vi du nhu hogt dgng ciia gen abcc2, protein lien kit kun loai... nhung khi tiep xiic vdi Zn d nong do cao dugc ddi hdi tieu hao nhiiu nang lugng de giii ddc co che vi chdng Iai si/ sn«ss oxi hda [9]
Tdm lai, ndng do Zn ttong mdi ttudng cing cao cang lam giim ti IS sdng cua au ttung^ca ngua van giai doan tit 1 - 7 ngiy tudi. Da xac dinh duoc ndng dd Zn giy chit 50 /<, au trung ci qua timg ngiy khio sit. Nhip tim vi kich fliudc iu ttiing ca Ngua vJn giam tuyen tinh theo su tang din cOa ndng do Zn khio sit.
3.4. Anh hudng eia Zn tin stf xuit hifn cic di lgt cua iu trung ci Ngtfa vin Au ttiing ca Ngua vin dugc giy nhiSm Zn IiSn tuc sudt tit giai doan phdi da xuil f" didang „ cac ca quan. Quan sit hinh thai cua 150 iu ttung ca Ngua vin d rail nong dp khao sat dudi kinh hien vi soi ndi vi kinh hiin vi dio nguoc, chiing tdi di ghi nh^ cac dang bat fliudng ttong hinh tfiii ciia iu ttiing d timg ndng dd khic nhau trong wm gim doan au ttung. Mdt sd dang bat tfiudng ttong qua ttinh phat triin cOa iu triing
bao gom: bien d?ng cdt sdng, phii ne mang ngoii tim (Hinh 2).
TAP CHi KHOA HOC BHSP TPHCM Nguyin Thi Thuang Huyin vi tgk
ITmh 2. Cac dgng di tat ddu trimg ca Ngua vdn (XIO).
mi ten chi vi tri di tdt,- A, B: di lgt cgt sing: C: di lgl cgt sing va phi tim rtii J '"^l "^^i^Z'f *"•' '*' **' " ' " ^" '^•^•"' *"" '••'""' '''""•• li"> rd. I u^ iiliicn .u uc che tong hgp DNA do tfiua kem cd flie li ldi giai tfiich cd khi ning nhii Su ^ ^ n^
m « ^ ' i " "^ ' T ^ ""P ™'^- ^"^ '^"^ '^"« "-W DNA da duac chung m i T l i T ^ T ' ^ •^"" " " r *'^ ™ "' * "^^ 0"^ *ai dddng vat. Rd ling S kem
^nguyen nhan gay ra qua, fliai d ddng vat cd vii, cdn fliira kem lai it giy quii tfiai l ^ r d^u nay CO tfie do co che bio ve cua gan vi cic mettillofliionein ttoni Z u ft i c^a con me. Trong kh, ci Ngua van li loii de tning, vi tfii phdi vi iu ttung s r ^ d n g ^ h T dugc su bio ve ttr ca me. [9, 11 ] "" " ung se xnong nhjn 4. Kit luin
^ Trong cic ndng do Zn khio sat, ndng do 1 mg/L li ngudng inh hudng din ti le song cua au ttung. Qua mdi ngiy nudi, ti le sdng cua iu ttung' gii^^ di 0,96nL '
• Trong cung mgt ndng do khio sit, ngay t,i ngiy tfiu 1 di cd inh hudng rd ret din h , l f ^ ' " . " " ? """^ ".*" ^ ^ '• '^ ^™8 '^"^ ^" '™ng -^a Ngua vin tfieo inh xic dinh dugc gia tt, LQ. giy chet 50% au ttimg tii ngiy fliu I din ngiy tfiu 7 ^' cua nif'fd-'7 't.'^"" * ' ' ™ . ' " ' ^ « '* ''«^'' ' ^ Siam ttiyin tinh tfieo sugia tang cua nong dg Zn Wiao sat vi tfidi gian nudi. O ngiy fliu 7, nhip tim vi kich thudclu S " ±0 O^m'" " * "™^ " ' ^" ^ ' " ^ ™ «'* "^ ''''•"> - 2.2^ - ^ ' P ^ h "
TAP CHl KHOA HOC DHSP TPHCM SS 6(84) nam 2016
TAI LIEU THAM KHAO
1. Bg Tii nguyen va Mdi trudng (2015), (hiy chuin kT Ihugl Quic gia vi chil luang nuac mat, QCVN 08-MT: 2015/BTNMT, H i Ndi.
2. Huynh Trung Hai va Trin Le_ Minh (2013), "Nghien cilu xd Ii niken, kSm, ddng vi chi ttong mdi tiudng nittat bing vd Ngao", Hot nghi khoa hoc lin thi 20, Trudng Dai hgc Bich khoa Hi Ndi, tt.66 - 72.
3. Begum, A., Harikrishna, S., & K.I. (2009), "Analysis of Heavy metals in Water Sediments and Fish samples of Madivala Lakes of Bangalore, Kamakata", International Joumal ofChemTech Research, Vol. 1 (2), 245 - 249.
4. Bnin, N.R., M. Lenz, Wehrii, B., & Pent, K. (2014), "Comparative effects of zinc oxide nanopaiticles and dissolved zinc on zebrafish embryos and elcufliero-embryos:
importance of zinc ions". Set Total Environ, 476-477: p. 657-66.
5. Chen, T.H., C C . Lm, & P.J. Meng (2014), "Zinc oxide nanoparticles alter hatching and larval locomotor activity in zebrafish (Danio tetio)", J Hazard Mater 277- 134- 40.
6. Hwang, P.P., Lin, S.W., & Lin, H.C. (1995), Different sensitivities to cadmium in Itlapta larvae (Oreochromis mossambiciis. Teleosleij, Arch, Environ Conlam Toxicol, 29, 1-7.
7. Jardine, D., & Litvak, M. K. (2003), "Direct yolk sac volume manipulation of zebrafish embryos and tfie relationship between offspring size and yolk sac volume"
Journal of Fish Biology, Vol. 63(2). 388 - 397.
8. Lawrence Christian (2007), "The husdbandry of zebrafish (Danlo rerio)- A review"
Aquaculture, 2 6 9 , 3 8 1 - 9 1 . ' 9. Long Y., L.Q., Zhong S., Wang Y., & Cui Z. (2011), "Molecular characterization
and functions of zebrafish ABCC2 in cellular efflux of heavy metals". Camp Biochem Physiol C Toxicol Pharmacol, 153(4): 381-91.
10. Roch, M., McCarter, J.A., Maflieson, A.T, Clark, M.J.R., & Olafson, R.W (1982), Hepatic meuillotfiionem in rainbow trout (Salmo gairdneri) as an indicator of metal pollution in the Campbell River system", Canadian Journal of Fisheries and Aquatic Sciences, 39, \596-60\.
11. Westerfield, M. (2007), The zebrafish book A guide for Ihe laboratory use of zebrafish (Duma rerio). Eugene, University of Oregon Press. Paperback.
IN^y Tda soan nhin dugc ha/: 15-4-2016: ngiy phin bi^n mnh gii: 23-4-2016- ngiy ohip nhin mng: 13-6-2016)
