52 2 厦门大学学报 ( 自然科学版 ) Vol52 No2 2013 3 JournalofXiamenUniversity (NaturalScience) Mar2013 doi:106043/jissn0438-0479201302025 邬阳 牛素芳 金媛 毛勇 * 苏永全 ( 361005) : (Cryptocaryonirritans) (Pseudosciaenacrocea) : 1574 mg/l 2h : ; : Q9589 : A : 0438-0479 (2013)02-0281-08 (Pseudosciaenacrocea) Wright [5] 2002 18SrRNA 2010 858 t [1] (ClassProstomatea) (OrderProreodonisa) : (FamilyCryptocaryonidae) (Cryptocaryonirritans) (Genuscryptocaryon) [2] [6] [7] (trophont) Sikama [3] 1937 (protomont) (tomont) (theront) Brown [4] 1951 : 4 [8-9] [10] CryptocaryonirritansBrown : (ClassOligohym- enophorea) (OrderHymenostomatide) :2012-07-12 : ( ) (200903029); (2011BAD13B08); ( 1222013-2) * :397263927@qqcom [11-12]
282 ( ) 2013 27 3~5d 14 141 142 1-20 11 / DNA [13] 045 rdna ITS (5 -GTTC- μm 121 30 CCCTTGAACGAGGAATTC-3 ) (5 - min 100IU/mL 100μg/mL TTAGTTTCTTTTCCTCCGCT-3 ) [14] PCR : 50μL 10 ( Mg 2+ )5 1mL 999 ml (1049mg/L) μl (10μmol/L) 1μLdNTPs(25mmol/L)1 1h μltaq 25UDNA (100ng/ μ L)1μL 12 PCR :94 5 min;94 30s55 30s72 1min 140 35 ; 72 5min [(159±33)g(107±05)cm] 7 DNA 1% ( 35 m 2 (n=20) )TBE DNA 1 m (12 ( ) L/min) 3~4d CLUSTALX [14] (27±1) 30±1 (GenBank (Trachinotusovatus) DQ270015) MEGA [15] : Kimura (Boot- [9] strap)1000 15 1000L [9] 3% ( ( 5 22:00~24:00) ) (28±1) 8000 / 3h 29±1 60h 13 (23L) (150 mm) 2011 6 10 27 3 (150mm) 1h (3~5d) 2
2 : 283 27 7d 50 ml (40 ) 100μL 5 96 ; (Shengyou Biotechnology CoLtd) 10μL 3 3 : 16 161 7 35 m 2 / 1m 140 7 20 21 (23L) 211 05251049 1574209831474196 mg/l 48h ( 1) 7 100 (LC 50 ) Reed-Muench [16] ( 2) 278 2482448h (TLm) ~609μm (21~40) μ m (50~70) μ m LC 50 : P1-50% TLm =C 1 + (C 2 -C 1 ) P 1 -P 2 :C 1 C 2 50% P l P 2 (Sc) : = - 100% 300 2 Sc = 03 TLm48 (TLm 24 /TLm 48 ) 2 TLm 24 TLm 48 2448h LC 50 2 162 1 96 Fig1 CirritansparasitizedinepidermisofPcrocea 150μL 500/ 0525 10491574209831474196mg/L DAPI 01248h (100 ) 4 U ( 3) [17] 3 3 163 3d 400μm 212 24 300 PCR ( 4) 2 ml 0 785bp Blast 52510491574209831474196 mg/l NCBI 2412h
284 ( ) 2013 (a) ;(b) ;(c) ;(d) 2 Fig2 Lifehistoryoftheparasite rdna 5 22 221 ( 1) ( 1049 mg/l) ; 1574mg/L 8h4 ; ; 4196mg/L 4 h 20 248 2448h LC 50 3672268122291836 1526mg/L 318mg/L( 2) 222 0~4h
2 : 285 50% 5 rdna Fig5 PhylogenetictreeforisolatesbasedonsequencesofrDNA [5] 8h 01248h 5 01248h Tab2 LC50ofaceticacidtojuvenilePcrocea (h) 2 4 8 24 48 4196 LC50(mg/L) 3672 2681 2229 1836 1526 3147157415741049 mg/l( 6)525 mg/l 8h IBM SPSSStatistics20 01-0819 : 2 LC50 1 Tab1 ThetoleranceofinfectedjuvenilePcroceatoaceticacid /% / (mg L -1 ) 0h 1h 2h 4h 8h 24h 48h 525 100 100 100 100 100 100 100 1049 100 100 100 100 100 100 100 1574 100 100 100 100 80 65 45 2098 100 100 85 75 55 35 35 3147 100 90 65 30 15 5 5 4196 95 75 35 0 0 0 0 0 100 100 100 100 95 95 95 6 Fig6 TimeneededtokilalCirritanstherontsbyaceticacid 4196 mg/l 5 min ( 7) 223
286 ( ) 2013 7 Fig7 ThemorphologicalchangeofCirritanstherontskiledbyaceticacid 525mg/L h 12h ( 3) 005 LSD 4h 4196 mg/llc 50 1238 mg/l; 12h 3147mg/LLC 50 64mg/L : y=4476+ 0219a+2095b(y: a: b: [18] ) 0802 0776; F 30424 005 3 Tab3 KilingefectofaceticacidonCirritanstomonts / (mg L) /% 2h 4h 12h 0 5 2 3 525 13 32 14 1049 20 36 60 1574 27 58 73 2098 43 88 95 3147 84 98 100 4196 94 100 100 0219 22:00~ 3:00) 2095T 7406 2449 p 005 : 3 Colorni Sun [1719] 4 ; ( ) 3~5d ( ; 2412
2 : 287 [4] BrownEAnewparasiticprotozoanthecausalorganism ofawhitespotdiseaseinmarinefishcryptocaryonirri- [20] 20982623mg/L tansgenandspn(agendaandabstrscimeetzool [21] SocLondon1950)[J]ProcZoolSocLondon195111: 1752~ 1-2 2623mg/L [5] WrightA D GColorniATaxonomicre-assignmentof Cryptocaryonirritansamarinefishparasite[J]Europe- Hirazawa [22] 315630 anjournalofprotistology200237(4):375-378 mg/l 2h [6] DigglesB KAdlardR DIntraspecificvariationinCryp- tocaryonirritans[j]journalofeukaryoticmicrobiology 4196 mg/l 199744(1):25-32 1574 mg/l (2h) [7] ColorniA BurgessCryptocaryonirritans Brown1951 100% thecauseof whitespotdisease inmarinefish:anupdate [J]AquarSciConserv19971:217-238 [8] DickersonH 4196mg/L WIchthyophthiriusmultifilisandCrypto- caryonirritans(phylum Ciliophora)[J]FishDiseasesand Disorders20061:116-153 [9] DanX MLiA XLinX TetalAstandardizedmethod ; topropagatecryptocaryonirritansonasusceptiblehost [20] 4h pompanotrachinotusovatus [J]Aquaculture2006258: 4196mg/L 127-133 (1574 mg/l) (2h) [10] LuoXCXie M QZhuX QetalSomecharacteristics ofhost-parasiterelationshipforcryptocaryonirritansi- solatedfrom SouthChina[J]ParasitolRes2008102: 1269-1275 [11] PironetF NJonesJBTreatmentsforectoparasitesand diseasesin captive Western Australian dhufish [J] AquacInt20008:349-361 [12] YoshinagaTIm H HNishidaSetalInvitroandin vivoeficaciesofionophoresagainstcryptocaryonirri- tans[j]aquaculture2011321:167-172 [13] SambrookJRusselD W [M]3 : 2002: 463-618 [14] ThompsonJ DGibson T JPlewniak FetalThe clustal X windowsinterface:flexiblestrategiesformul- tiplesequencesalignmentaidedbyqualityanalysistools [J]NucleicAcidsResearch199725:4876-4882 [15] TamuraKDudleyJNeiMetalMEGA4:moleculare- : [1] 2011 [M] : [16] ReedLJMuench HAsimplemethodofestimatingfif- 2011 typercentendpoints[j]theamericanjournalof Hy- [2] giene193827:483-485 [J] 200936(1): [17] ColorniAAspectsofthebiologyofCryptocaryonirri- 4-7 [3] SikamaYPreliminaryreportonthewhite-spotdiseasein marinefishes[j]suisan-gakukai-ho19377:149-160 volutiongeneticanalysis(mega)softwareversion40 [J]MolBioEvol200724(8):1596-1599 tansandhyposalinityasacontrolmeasureincultured gilt-headseabreamsparusaurata[j]disaquatorgan 19851:19-22
288 ( ) 2013 [18] [22] HirazawaNSyun-ichirouOshimaTakashiHaraetal [J] 200832 Antiparasiticefectofmedium-chainfatyacidsagainst (1):13-18 theciliatecryptocaryonirritansinfestationinthered [19] Sun H YZhuX QXie M QetalCharacterizationof seabreampagrus major[j]aquculture2001198:219- Cryptocaryonirritansisolatesfrom marinefishesin Ma- inlandchinabyitsribosomaldnasequences[j]para- sitolres200699:160-166 [20] [J] eggsandoncomiracidia andtheciliratecryptocaryon 200535(1):45-46 irritanscystsandtheronts[j]aquaculture2003223: 228 [21] [J] 1-13 19801:37-38 [23] HirazawaNGotoTShirasuKKilingefectofvarious treatmentsonthemonogeneanheterobothrium okamotoi ObservationontheLifeHistoryofCryptocaryonirritansinEasternSeaof FujianandResearchonAntiparasiticEffectofAceticAcid WU YangNIUSu-fangJIN YuanMAO Yong * SU Yong-quan (ColegeofOcean & EarthSciencesXiamenUniversityXiamen361005China) Abstract: Cryptocaryonirritans aholotrichousmarineciliateprotozoanthatcausing whitespotdisease isoneofthemostdevas- tatingparasitesof marinefishin Fujian ProvinceInthepresentstudyastrainofCirritans-likeparasite wasisolatedfrom Pseudosciaenacrocea aquicultureareasineasternfujianprovinceandthenidentifiedthemorphologicalfeaturesandbiochemical characteristicsoftheparasiteswereidenticaltothosecirritansstrainswhichhadbeenreportedinotherplacesthisfactsuggested thatthesampledpathogenwascirritansinadditiontheacutetoxicityofaceticacidtojuvenilepcroceaandcirritanswasstud- iedtheresultsshowedthatcirritanstherontsweremoresensitivetoaceticacidthantomontsmeanwhilethesusceptibilityoface- ticacidtojuvenilepcroceawasanimportantfactorwhichwasconsideredintheprocessofkilingcirritanstheresultsindicated thatdealt whitespotdisease infectedpcroceawith1574mg/laceticacidfor2hcouldkilcirritanstherontsefectivelyaswel asunharmfultothehostsitwasasafedrug-usingstrategyforpreventingandcuring whitespotdisease Keywords: Pseudosciaenacrocea ;Cryptocaryonirritans;aceticacid