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Tap chi Khoa hoc va Cdng nghe biln T9 (2009) So 1 Tr 55 68 DANH GIA ANH HUOfNG CUA DAU DIESEL (Pha hda tan) LEN HAI LOAI TAO Chaetoceros muelleri va Skeletonema costatum HOANG TRUNG DU Tdm tdt Nhung d[.]
Tap chi Khoa hoc va Cdng nghe biln T9 (2009) So Tr 55 - 68 DANH GIA ANH HUOfNG CUA DAU DIESEL (Pha hda tan) LEN HAI LOAI TAO Chaetoceros muelleri va Skeletonema costatum HOANG TRUNG DU Tdm tdt: Nhung dnh hudng cua ddu diesel (WSF- pha hda tan) len lodl tdo Chaetoceros muelleri (C muellerij vd Skeletonema costatum fS costatum^ duac xem xet qua cdc thi nghiem kiem dinh dgc Id Ket qud da chi cdc mice dnh hudng cua tieng ndng ddu len cdc chi sd nhu mat te bdo chlorophyll a, vd ndng sudt cua hai lodl tdo phd bien vuc nudc ven bd Sic dung chi sd kiem dinh de ddnh gid mice dgc tinh cua ddu cho thdy ddi vdi tieng loai tdo khdc se chiu dung a cdc mite dnh hudng "stress " ciia ddu diesel ciing khdc Cdc chi sd ve ddc tinh ciia ddu gdy su ice che tdc phdt trien cua tdo nhu: vdi IC25 Id 224.8/.igWSFA ±42.4, ddi vdi Idal tdo C muelleri va 454pgWSF/l±I8 luang ieng ddi vdi tdo S costatum Trong dnh hudng ciia ddu gdy ice che 50% (IC50) ddi vdi cd hai loat tdo deu > 500pg/l Anh hudng ciia ddu qua cdc chi sd sinh thdi nhu hdm lugng chlorophyll a, ndng sudt, ciing chi su suy gidm ticang ieng vdi hdm lugng nhiem bdn ddu tdng Mite suy gidm 50% so vdi ddi chiing bdt ddu xdy lie ndng 125/jgWSF/l trd len vdi tdo C muelleri vd 250 pgWSF/l vdi tdo S costatum Ddng thai vdi nhung yeu diem ve mat sinh thdi hgc, chlorophyll a dugc xem xet dua vdo sic dung kiem dinh ddc td ciia cdc chdt hda hgc tren tdo.vd cd khd ndng ieng dung cho viec ddnh gid nhung viing nudc cd ddu hieu nhiem bdn ddu, nhdm cdnh bdo sdm nguy ca sinh thdi a cdc viing nicdc ven bien bl de dga nhiem ddu Key word: Kiem dinh dgc td, ice che, chlorophyll a, ddu diesel I Md DAU Nhiem ban dau thai la mgt thue te mang tinh toan cau, chiing bat ngudn tii nhung nguydn nhan: Giao thdng van tai va van chuyen dau - Khai thac tham dd dau khf ngdai khoi - Cac hgat ddng khac tren bidn nhu xu ly, phan phdi va kinh doanh dau, danh bit ea, du lich - Cac hgat ddng ed lidn quan den sii' dung dau cac luu vue, thai vao sdng va dugc sdng tai bidn Viet Nam dang dung trudc su de dga cang gia tang eiia tinh trang d nhiem dau nude bien eac boat dgng nhgp nhjp va rao riit d cac cang biln va van tai bidn trang nude va Qudc td, viec sii' dung eae san pham dau md va cac hoat dgng tham dd, khai thac ngoai khai Nhin ehung ham lugng hydrocarbon diu vuc nudc ven bd cao hon d ngdai khai (FAO, 1982, Nguyin Minh Son va cdng su, 1999) 55 Cac phan hda tan ciia dau thd va eac san pham tinh che ddu cd cac ddc tinh tiiy thudc vao tinh chit va miic eua tiing loai ldn thuc vat va ddng vat bidn Dau nhien lieu thu'dng dugc su dung dl boat dgng eac dgng ea may (thudng la dau diesel), chiing cd chiia cac hydrocarbon tham va cae hydrocarbon thudng dugc xem la phin diu ed doe cip tinh, ddng thai chiing cung cd cac anh hudng ddc man tinh, va dugc coi la chit cd kha nang gay ung thu Do dgc eiia dau diesel phu thude vao sd lugng vdng thom ciia chiing va cae chit boat ddng bd mat (vdi nhung nhdm hydrophobic nhu alkyl phenol va R2CHOH la cd ddc td cao) Mgt so nha khoa hgc da thuc hien nhung nghien eiiu ve anh hudng eiia d nhiem dau cho ring he sinh thai bien ven bd la nai cd nhay earn nhat ddi vdi viec d nhiem dau nhu: cua sdng, viing dat ngap nudc (rung ngap man, bai tridu, ), ran san hd, va tham cd biln (GESAMP, 1993, Patin, 1982) Nhung anh hudng sinh hge eua cac dau thai phu thudc vao Igai dau va thdi lugng tidp xiic tdi cac he sinh thai tu nhien Chat d nhiem gay ddc cho sinh vat va lam bien ddi dac diem mdi trudng sdng; Ci ngudng gay dgc cd the giet chet sinh vat mgt thdi gian ngan; d dudi nguong thi lam giam kha nang hgat ddng sdng va kidm mdi nhiem dau nude cd anh hudng ddc tfnh manh ldn thuc vat phu du (UNEP/EAS, 1996) Vdi ham lugng dau d nhidm cao lam giam tdc sinh trudng, kha nang sinh san va tap doan hda ciia san hd ciing nhu tae ddng tidu cue den hoat dgng bit mdi va bai tiet (Loya and Rinkewieh, 1987, Grigg and Dollar, 1990) Nghidn ciiu anh hudng nhiem ban dau trdn sinh vat dieu kien phdng thi nghiem la nham xae djnh tac dgng cd hai ciia dau thai len sinh vat thi nghiem, vdi cac phuong phap thi nghiem va xu ly phu hgp (Malallah et al., 1996; Barron et ah, 1999; Long et al., 2003; Adebayo, 2005) Ket qua phdng thi nghiem se lam ca sd khoa hgc ling dung giam sat sinh hgc (biomonitoring) Da cd mdt sd nghidn eii'u da diing cae chi sd nhu chlorophyll huynh quang nhu la ehi thi anh hudng eua cae hydrocarbon thom (Christopher et al, 2001), cae kim loai nang (Maefarlane va Burchett, 2001) cae thudc tru sau diet cd ldn su phat trien eiia thue vat (Berard et al, 2004) hoac sii' dung sic to thuc vat viec djnh lugng sinh khdi va tdc phat triln eiia cac vi tao (James et al, 2001) Qua bai bao chiing tdi vdi muc dfch nghien eiiu bien ddng ciia mdt sd chi sd sinh thai nhu ham lugng chlorophyll a, mat tl bao, nang suit sa cip loai tao nghien eiiu duge tidp xiic vdi mdi trudng dau hda tan nude biln Vdi ky thuat kidm dinh doe td duge sii dung nhu la mdt phuong phap danh gia nguy ca sinh thai cua dau ddi vdi tao 56 II TAI LIEU VA PHUONG PHAP NGHIEN CUtJ Mau sinh vat - bd tri va thuc hien thi nghiem Tao dung thi nghiem Gidng tao Chaetoceros muelleri va Skeletonema costatum dugc nudi giir gidng tai phdng tao thuoc Trung tam nghien cuu nudi trdng thuy san va phdng Sinh vat phii du Vien Hai duong hgc Sau dd tao dugc nudi sinh khdi de dat mat ban dau la tu 10^10*^tb/inl (De su dung eac thf nghiem mat tao ban dau la 10''tb/ml) Tao dung de kiem dinh dugc cay chuyen lien tuc de giii gidng dugc mdi va dugc kiem tra trudc su dung vao thi nghiem Ddi vdi tao silie ndi chung, S.costatum va C muelleri ndi ridng ba Igai mudi dinh duong ddng vai trd quan trgng dd su tri sinh trudng, phat trien cua tao la: Nitc, phdt pho, silie Thanh phan mdi trudng bao gdm: - Dung dich 1: NaNOj, NaH2P04H20, MnCl2.4H20, FeCl3.6H20 - Dung djeh 2: ZnCb, C0CI2.6H2O, CaS04.5H20 Dung dich gdc eiia dau hda tan nude bien dugc tao bang viec sii' dung dau diesel nhien lieu dugc khuay manh nude bidn da dugc Igc qua mang Igc 0.7p,m vdi tdc khoang 1000 vdng/phiit, thdi gian la gid, sau dugc tach lam pha hda tan va khdng hda tan - lay pha hda tan {con goi la Water Soluble Fraction- WSF) dd dimg thi nghiem Thi nghiem duge thuc hien phdng thi nghiem kiem djnh doe td tai Phdng Sinh thai va Mdi trudng Bien, Vien Hai duang hge Thi nghiem duge thue hien dieu kien nhiet phdng dn djnh (nhiet = 26 ±1°C), anh sang dugc eung cap bang den huynh quang vdi cudng sang la 3800 lux va dugc chidu sang lien tuc 24/24gid' Cac Id thf nghiem dugc thuc hien vdi mau song song (replicate) tai mdi ndng khac nhau, thdi lugng tidp xiie ciia sinh vat ddi vdi dau la 96 gid va tat ca ddu dugc cung cap mgt lugng dinh duong ban dau nhu Ngoai cdn thue hien mau ddi ehiing vdi nude bien sach va thuc hien theo qui trinh thi nghiem ciia ASTM, 1994 Cac yeu td nhiet do, pH, man va oxy hda tan dugc true tiep bang may YSI tai cac ndng kidm djnh Thai gian thf nghiem tren loai tao la 96 gid, mau d cac binh duge lay dd xae dinh eae thdng sd: - Ham lugng hydrocarbon dau thuc td dugc xae djnh trudc va sau cac dgt thi nghiem, miu dugc chiet dung mdi n-hexan, sau dd duge tren may sic ky khf HPGC 6890, vdi dau dd FID 300°C, cdt sac ky mao quan HP-1, chay tren chuong trinh nhiet (US EPA, 1996) 57 - Chlorophyll a, b, c: Mau dugc lac deu va Igc qua mang Igc Wathman GF/F, sau dd dugc chiet dung mdi Aceton 90%o va tren may quang phd UV/Vis va tinh theo cdng thii'c ciia Parson e? a/(1984) - NSng suat sa cap: sii' dung phuong phap C''*, duge tren may nhap nhay ldng (Liquid Seintilation Counter) ( Grassholf, 1994) - Mat tao: Sau 96 gid thf nghiem va mau dung de tfnh mat tao dugc thu vdi lan lap / binh, mau dugc cd djnh bang Lugol 5%, sau dd dugc dem trdn kinh hien vi d vat kinh 40x, vdi budng ddm hdng cau Xu ly sd lieu Anh hud'ng cua dau diesel len tdc sinh trudng ciia tao dugc tfnh theo cdng thuc: % uc chi (inhibition) = (Uck - Utox)*100/Uck Trong dd: Utox la tdc sinh trudng d tung ndng cd hien dien dau diesel thi nghiem Uck la tdc sinh trudng cua tao d binh ddi chiing Anh hudng uc che (%>) ciia dau len lugng chlorophyll a dugc xae djnh theo edng thiic (Fargasova, 1997): %f'' = (l-c/Kchl)*100 Trong do: c la lugng chlorophyll a tai ndng dau tiep xiie; Kchl la lugng chlorophyll a d Id ddi chung Sd lieu duge thdng ke va phan rich trdn phan Scienrific Sofware, MeKinlayville, CA) cac chi sd Effects Concentration) va LOEC- (Lowest Observed dch cung vdi 1C25, IC50 (Inhibitory Concentrarion d cua dau anh hudng len su phat tridn ciia loai tao mim Excel va ToxCalc (Tidepool kilm djnh NOEC- (No Observed Effects Concentration), dugc phan 25% va 50%), de danh gia dgc tfnh III K£T QUA VA THAO LUAN Tu ket qua ehi riet vl mat te bao (bang la, b, e va 2a, b) chiing tdi nhan thiy ring anh hud'ng ciia dau len su phat trien ciia tao la su lic chi tdc sinh trudng ciia tao, vdi thdi lugng tilp xiie d 96gid thi su lic chi > 10% (so vdi ddi ehiing) len su phat triln eiia tao C muelleri xay tai ndng tu 125pgWSF/l trd len (bang 3) Trong dd su dung phin mdm tfnh toan Toxealc, kit qua ehi ring ndng diu khdng gay anh hudng (NOEC) la 31.25|agWSF/l va ndng diu thip nhat gay hieu ling (LOEC) la 62.5)MgWSF/l toan bg cac dgt thf nghiem tren tao C muelleri; Ndng diu gay li'c 58 chi len tdc phat triln cua tao d 25% (IC 25) trung binh la 224.8p.gWSF/l ±42.4(dao ddng tu 176 - 269^g/l) va ndng gay uc chi d 50% (IC 50) deu > 500ngWSF/l (bang 4) Bang la Thi nghiem dgt 1: tii 5.8.2004 - 9.8.2004, Tren loai tdo: Chaetoceros mulleri So lan lap N=3' Mat tao (tbxlO'/ml) Chl-a mg.m"' N/Suit mgC/m^/h Trung binlr Stdev 117.3 395.4 17.9 288.1 33.0 C31.25ng/L Tmng binli 89.7 233.8 377.2 Stdev 12.7 129.5 C62.5|.ig/L Trung binh Stdev 83.0 4.6 57.0 4.0 35.7 4.2 15.0 2.0 203.1 Ky hieu Doi chiing Cl25ng/L C^SOng/L CsOOjig/L Trung binli Stdev Trung binh Stdev Trung binh Stdev 11.7 184.5 59.9 227.5 26.5 136.0 87.4 281.2 273.0 344.1 91.0 *Stdev; Do lech chudn Bang lb Thi nghiem dgt 2: hi' 19.8.2004 - 23.08.2004, Tren loai tao: Chaetoceros mulleri Ky hieu Ddi chung C3|.25|.ig/L C62.5|ig/L Cl25ng'L C250ng/L CsOOHg/L So lan lap N=3' Trung binli Stdev Trung binh Stdev Trung binli Stdev Trung binh Stdev Tnmg binh Stdev Trung binli Stdev Mat tao Chl-a N/Suat (tbxl04/ml) mg.m-3 mgC/m3/h 129.3 252.0 500.59 7.1 18.1 108.3 222.5 434.39 5.1 18.7 96.7 116.3 146.58 4.0 6.0 65.0 130.8 136.59 2.6 61.0 41.0 83.0 152.45 4.0 9.4 18.0 65.2 89.83 3.0 9.4 *Sldev; Dd lech chudn 59 Bang le Tlii ngliiem dgt 3: ti:r 20/09 din 24/09/2004, Tren loai tao: Chaetoceros mulleri Ky hieu Doi chiing C3l.25|.ig'L C62.5(ig/L C|25jig/L C250ng/L CsOOng/L So lan lap N=3" Mat tao (tbxlO'/ml) Trung binh Stdev Trung binh Stdev Trung binh Stdev Tnmg binli Stdev Tmng binh Stdev Tmng binh Stdev 106.3 18.9 101.3 10.2 75.3 7.5 58.0 4.6 24.0 4.0 11.3 3.2 N/Suat Chl-a mg.m"'' mgC/m'/h 275.0 89.4 228.3 8.6 135.5 2.9 138.6 22.8 84.9 13.7 38.9 14.8 279.29 334.39 180.69 187.99 110.59 34.67 *Stdev; Do lech chudn Bang Id Thi nghiem dgt 4: tu 04/10 den 08/10/2004, Tren loai tao: Chaetoceros mulleri Ky hieu Ddi chiing C3l.2S|.ig/L *^62.5).ig/L Cl25Mg/L C250Mg/L CsoOMg/L So lan lap N=3" Trung binh Stdev Tmng binli Stdev Trang binh Stdev Tmng binli Stdev Trung binh Stdev Tmng binh Stdev Mat tao Chl-a (tbxloVml) mg.m"' 144.3 292.3 7.5 53.2 109.C 186.1 11.4 53.0 161.4 87.3 3.5 62.2 54.0 79.5 6.2 5.3 68.3 32.3 4.5 14.5 14.0 58.7 6.0| 8.6 N/Suat mgC/m'/h 192.23 207.78 188.23 105.61 107.17 52.41 *Stdev; Do lech chudn Cac gia tri tfnh toan ve % tic che su phat trien ciia tao S costatum, cho thiy % uc ehi >10% chi xay a nong dau la 500|MgWSF/l Ci nong nho hon hau 125 j.igWSF/1 thi su lic chi phat trien la khdng dang ke, dieu cd khac so vdi tren tao C.muelleri Chi so tfnh toan vl dgc tinh eiia dau vdi phan mdm Toxealc cho thay: chi sd NOEC la 31.25f.igWSF/l va LOEC la 60 62.5|ugWSF/l tuong tu nhu anh hudng eua diu ddi vdi tao C muelleri Tuy nhien, chi sd IC25 lai cao hon rit nhilu, trung binh la 454 )agWSF/l ±18 (khoang tu 437 - 473p.gWSF/l) Dilu cd thi thiy nguong chiu dirng "stress" cua tao S costatum gay bdi diu cao hon tao C muelleri Bang 2a Thi nghiem dgt I: tu 24/10 din 29/10/2004, Tren loai tao: Skeletonema mulleri Ky hieu Ddi chiing C31.25ng/L C62.5ng/L Cl25^Lg/L C250ng/L CsOOng/L Ddi chiing N/Suit Sd lan lap Mat tao Chl-a (IO"* tb'/ml) mg.m"^ mgC/m^/h n 364.7 82.3 135.3 Trang binh 5.4 Stdev 7.0 220.0 113.0 95.5 Trung binh Stdev 4.4 21.4 Trung binh Stdev Trung binh Stdev Trung binh Stdev 114.3 7.8 99.3 5.1 81.3 8.0 37 36.7 4.5 100.6 25.0 65.2 8.1 78.3 10.9 59.4 59.2 0.9 Tning binh Stdev Stdev 241.5 262.2 213.8 186.9 Bang 2b Thi nghiem dgt 2: tir 29/10 den 03/11/2004 Ky hieu So Ian lap n Mat tao (lO" tb/ml) Chl-a mg.m"^ N/Suit mgC/m^/h Ddi chiing Trang binh 116.7 192.7 Stdev 5.0 50.8 7.5 Trung binh Stdev 108.7 79.1 22.1 172.9 4.0 Trang binh Stdev 97.0 63.0 144.8 1.4 14.2 Trang binh Stdev Trung binh Stdev Trang binh Stdev 99.0 4.2 85.7 4.7 26.7 4.0 122.5 4.5 78.4 11.4 13.3 9.3 C31.25Mg/L C62.5^lg/L Cl25ng/L C250ng/L CsOOng/L 161.3 196.0 20.7 61 Bang Kit qua tfnh ve % li'c chi tdc phat triln thi nghiem tren tao vdi thdi lugng tilp xiic la 96h vdi cac ndng dau khac C muelleri Dot Dot Dot Dot S costatum Dot Dot Dot 31.25mg WSF/1 5.34 3.30 0.79 5.03 62.5mg WSF/1 6.97 5.64 7.16 9.53 125mg WSF/1 14.88 13.84 12.77 19.25 250mg WSF/1 24.73 23.35 31.73 29.65 500 mg WSF/I 42.99 40.34 47.91 46.58 6.1 1.3 3.6 5.9 3.7 5.0 8.7 3.3 4.4 12.6 6.3 9.3 28.4 30.8 27.2 Bang Cac chi sd kiem dinh vd ddc tinh ciia dau len tao sii dung phan mem tinh tdan ToxCalc Dot thi nghiem C muelleri Dot Dot Dot Dot S costatum Dot Dot Dot Replicates Thoi lu'ong tiep xuc(gi6') 3 3 96h 96h 96h 96h NOEC 500 >500 >500 >500 >500 >500 Nhu'ng anh huang ciia dau len su phat trien ciia tao ed the xem xet qua cac ket qua chi sd sinh thai nhu: ham lugng sac td chlorophyll, nang suat, mat td bao tao Qua ket qua ciia cac dgt thi nghiem da chi rang cac chi sd cd sir suy giam tuong li'ng vdi ham lugng dau tang dan, lugng sac td ehlorophyll a tai eac thf nghiem ed thai lugng tidp xiic 96gid vdi cac ndng dau khac giam mgt each ro ret (hinh 1) Xu hudng tren cung thay ro d cac gia tri vd nang suat, ddng thdi tir cac sd lieu tinh phin tram suy giam cho thiy a miic 25% so vdi ddi chung xay bit diu tang tu ndng 62.5|ag/L.(hinh 2) 62 B Thi nghiem tren S.costatum costaium A Tiiinghiem tren C.muellen "g 150.0 -.-Dd ^_^^ 1;; ^ r r f x o ™° ^ SOO E |2 00 • \ E ~^Da\2 -6-Dol3 -B-TB -ãEtot b * ^ orDphyll a E ã"" -ô-Êtot2 -a-Dot Cr""^ ^ 500 ^ ^ ^ ^ ^~- ^^~a ^n.^ \ -o-TB ^ " ^ ^ ^ ^ i 31 25 • 62 00 125 250 31.25 SOO Nong dau(ug'L) 62.5 125 500 250 N6ng d4u(ug/LI) Hinh 1: Su suy giam cua chlorophyll a cac thf nghiem tren tao: (A) C muellen va (B) 5' costatum doi vai doc tinh cua dau diesel (B) Thi nuhii-m ibit 2{l'M3.(ffi.2()(IJ) iren laii ChmiliK: (A) Thi nuhiOm dr/t l(5-y.(IH.20a4) iK-n liia ChaHaaros m u d l c r i ^ I O : ^ 100.0 I 75 50.0 D 25.0 ; c r -• 0.0 ' "nCiiiori'pIiylia 100.0 - 75.0 : U 50.0 I 25.0 , ^ 0.0 31 81,3 131 181 231 281 331 361 431 ' 31.3 61.3 13-1 181 231 281 331 381 431 481 N I25pgWSF/L for C muelleri and > 250figWSF/L for S costatum algal species that It's was compared with controlled experiments At the same time, based on the advantage of chlorophyll a parameter It was examined to use In toxicity test for chemical poisons on algal testing and we recognized that it can be applied for assessing the waters with sign of oil pollutant Moreover, It's very useful in early warning ecological risk in the coastal waters where frequently occur all pollution Keyword: Toxicity test, Inhibition, Chlorophyll a, diesel oil Ngdy nlidn bdi: 26 - 10 - 2008 * -^ • Ngudi nltcin xet: PGS.TSKH Nguyin Tdc An 68 „• ,- x^ TT-T^ i Dia chi: Vien Hai Duong hgc ... ON TWO ALGAL SPECIES CHAETOCEROS MUELLERI AND SKELETONEMA COSTATUM HOANG TRUNG DU Summary: The effects of diesel oil on the algal species are carried on C muelleri and S costatum by using toxicology... sinh thai cua dau ddi vdi tao 56 II TAI LIEU VA PHUONG PHAP NGHIEN CUtJ Mau sinh vat - bd tri va thuc hien thi nghiem Tao dung thi nghiem Gidng tao Chaetoceros muelleri va Skeletonema costatum. .. tugng loai tao sii'' dung lam kiem djnh, nhu vdi tao S costatum va C muelleri la tao silie phan bd kha phd bidn d vimg bien ven bd Mac du ca hai loai tao ddu cd cimg ndng khdng cd hieu ung anh hudng