366 DOI:10.13350/j.cjpb.190326 +, BC - M,, 2,) (0 a( Y0 6 a( Y, M' V0, 6 26 H W, 200025),T6 =& 26 P 6, ",T6,T6,,O @, A,M D ) B 69 PW 6,,E W SB #$I O)9 + 9PW,T6 A, 3:, A,T6, +, +,T6; A; R383.2!"#$ A %& 1673 5234(2019)03 0366 05 [.2019 Mar;14(3):366-370.] 犚犲狊犲犪狉犮犺狆狉狅犵狉犲狊狊狅狀狊狆犲犮犻犲狊犻犱犲狀狋犻犳犻犮犪狋犻狅狀狅犳犉犪狊犮犻狅犾犪狊狆狆. TANG Meng ting,zhou Yan,CHENG Na,XU Xue nian ( 犖犪狋犻狅狀犪犾犐狀狊狋犻狋狌狋犲狅犳犘犪狉犪狊犻狋犻犮犇犻狊犲犪狊犲狊, 犆犺犻 狀犲狊犲犆犲狀狋犲狉犳狅狉犇犻狊犲犪狊犲犆狅狀狋狉狅犾犪狀犱犘狉犲狏犲狀狋犻狅狀, 犠犎犗犆狅犾犪犫狅狉犪狋犻狀犵犆犲狀狋犲狉犳狅狉犜狉狅狆犻犮犪犾犇犻狊犲犪狊犲, 犓犲狔犔犪犫狅狉犪狋狅狉狔狅犳犘犪狉犪狊犻狋犲犪狀犱犞犲犮狋狅狉犅犻狅犾狅犵狔, 犕犻狀犻狊狋狉狔狅犳犎犲犪犾狋犺, 犛犺犪狀犵犺犪犻 200025, 犆犺犻狀犪 ) 犃犫狊狋狉犪犮狋 TwotrematodespeciesofthegenusFasciola,FasciolahepaticaandF.gigantica,arecommonparasitesin thebileductsofcatle,sheepandothermammals.f.hepaticaandf.giganticasoresembleeachotherthatonecouldnot distinguishthembymorphologicalmethod,diferentiationofthesetwospeciesisoneofthehotspots.thispaperintends toelucidatetheprogressinspeciesidentificationbyusing modernbiotechnologysuchaschromosomeanalysis,protein polymorphismandmoleculargenetictechniques,revealgeneticdiversityofinter andintra speciesandrelationshipsbe tweenfasciolahepaticaandf.giganticainthegenusfasciola. 犓犲狔狑狅狉犱狊 Fasciolaspp.;speciesidentification;review,T6, T 6 M 5, K, M + 8, 240 M,T6,NM N 9100 [1], T 6,, 2.5 ~3 M, T6,* & = c H, _ M 30? [2], T 6 = W G, 2 6 $, T 6 \ - U,,,=,9,T6, T 6 H ", ) /,,T6 _,T6( 犉. 犮犪犾犻 犳狅狉狀犻犮犪 Sinitsin,1933), T 6 ( 犉. 犺犪犾犻 Sinitsin, 1933),,T6 ',T6( 犉. 犻狀犱犻犮犪 Varma, 1953) [3],T6, T 6, @,5 /,, M 6, M ) Y V <)/+ [4-8] / U,, T 6,/,, T 6? ) /, ) B/0 M -E ; ^ ', DNA 2 R,, T 6, "0 ",T6 [9-14] E, T 6,,T6,,, T 6, 4 1 ( :; Y, / 9, O, R 4 R,O, 6 B,_ D 4, 6 9 > ^, M),5, T 6, O @,?, O A /, T 6, T 6, H,,T6, A ) / b,,, T 6,,OR % >,, / > P 6*69, & :, ; S B 6 L, + 9PW,T6,,,T 6 A ^, 3: 1. SB U W> Y?, 3 V, H",-,0!E /B, T 6? C U,,T66 SB U 2n=20,,T6 SB U 3n=30,5/K \ ^ B,T6 [15] -E,T66 2n=20,3n=30,9 2n/3n 3, 0 9 B A [16] Itagaki [17] B DNA, K (ITS2) 7 -E B,T 634, Z 0 6, T 6,ITS2 2 ' 6 I T CD G(No.TDRC 22),E mail:xuxn@nipd.chinacdc.cn () `(1993-),,,, " # 6 E mail:tmt2015@126.com
367,, T 6 [18], T6 SB U @W-E, B 6B,H W U,, T 6 6.5 [19] / _ S,,T 6 0 B, 4 W B a #" [20] _, T 6 3 4 S B,, C B,,T6 B,T6 )>, E,= T, 6 3 L, E 5 C C 3LHB, 1988 a # " [21],, T6 SB S B ) W 0 b,,,,? SB./,T6 2 =2 2.1 34>5 7X 6B *K, =6, F+ \ X#$, DNA 2, X,GL ^ *, G L 7 X, 9 :\ L,H". D, PW 6 6 V L, a # " [20] ^ _,T6,T6 7X,X, X 7X>, A,T6, T ^, SB Z I 5 L6X,X [ >,,T6 2 `, X ',, T 6, 5 X' ` >, + 6, - Agatsuma [22] ^ - E, T 6, 12 7 X, 0? AK Got Gpi 6 Pgd Pgm 2 ) / +, 0,T6 ),H -E,T6 @,Got, T 6, 7X 9 \,T6, +,/ 0 AVP 2.2 :;67 ^ 7 X, ) # $ B #$I \ 6,+,, M) R ), I 6 J( 6 6 6 [23-25] Lofty [26] P 9, T 6 34 A,,O,T6, T6, )#$! @,5 / ph5.14-7.4 R,,,T6,T6 B Lee [27] W >, T 6, ) # $ ]=634, W ) XY Y,, T6 )#$ @,5 W,,T6 [ +,C 30 ; 3, >, T 6, R ) : \, [28] )#$ O) / A, T 6 V H ) C,5 " *,-. 3./, O L / P W, T 6,,5 9$, A Z ) B + 69 & :,/ 6, 0 A F,,,T6, ^ / U PW,T6 A, + I * DNA (genomedna),mtdna(mitochondrialdna, J B DNA) rdna(ribosomaldna, B DNA) $ DNA(micro satelitedna, DNA), 09rDNA mtdna, P rdna B RNA, DNA 2, 'H-, * 2, 3 H 3L&' 0 K K (ITS1 ITS2)+' 0 ' O), 3 L [29-31], R ) 9 H -? + W3,, A,/, T 6, A0 P J B * F @ A 9 Q, 0 S L X I(cytochromecoxidasesub unit1,cox1) 0 * X * X I (nicotinamideadeninedinucleotidedehydrogenasesubunit 1,NADl) PW 6, A,5,3L&')/+, P +,T634> ',3L,9 Z C, 3.1 + 4 8 9 : ; - A > 5 (poly merasechainrestrictionfragmentlengthpolymorphism,pcr RFLP) W6/3L 80* * G H H= @ L M,PCR RFLP 9 R 5 6 < U DNA, P Y) X, GL, <=6 \> + ' U, DNA, B, + O ) 3 4 A PCR RFLP 9 E DNA, _, - [32] < ^ /,,T6rDNA,ITS2, 5.8S 28S, (ITS2+ 2),P Y ) X Hsp92Ⅱ RcaⅠ 1 L, X,X ' )/ + 5. +, 9,T 6 2 D Z A / E =,T6, E,T6 EITS2 2, T 6, T 6, R,X Z ', C,T6 C )/R,".` ",T6,* `C L \ ` +, C 9, Q = C [33-35] ".` ", T 6,ITS1 2 ', T 6, T 6, H R, C 6 6, \ Shoriki [36] D ^, T 6, - * pepck ( QW X) pold(dnapolymerasedelta,dna Xδ) 2, pepck / 6, + 2.30% ~2. 68%,pold 4.93%~5.22%,5, + W 1.0%,* P *,,T6 E34 ^ PCR RFLP,X Z A 15 ".` ",T6* ^, T 6, ', A, T 6 P ITS1 V + 3 4 PCR RFLP,? 1/3 A,T6 A - * pepck pold 2 H-,ITS1 2 D,T6, 3.2 /0>5 DNA < (Randomamplifiedpolymor phicdna,rapd) 9P 26 T ( ` 8~10 )V 56? * 34 56 <, W> = > B G L M @ -.^56,, < Z_ W ]Q O) RAPD 9, B / 5 6 6 ) P),U P W 6,, J
368 6.a 6 T6 [37-41] Mcgarry [42] RAPD 9 R ) PCR, P 6 56,T6 E34 <, <=6,D Z78,T6,T6, R )56,9 G c,r ) PCR, T 6, T 6 / [43] / P,, T 6 E, &,,T6,;, R > [,, T 6, 6,T634 RAPD D,_ <!,,O>,, T 6, T 6, @ < 0 A, T 6 +? 0.255,, T 6 + 0.106 ~0.255,H +,,T6,T6+ 0.532, RAPD,T6, 7 3.3 >4 PCR (MultiplexPCR ) 1988 Chamberlain [44] H PCR b, : / A,,/ 6 9, PB-.< H PCR, / B 0 @2 9 R )56, < DNA >, 2,_! A6,+, \E, W < DNA U,, H PCR,9 ', B 0 \ 9 I ' ' " A, L 3 I B C, R ) 4 ' Thanh [45], T 6 mtdna, cox1 trnt rrnl 78,T6, T 6, R ) 5 6 P, 56,Gc^ H PCR /,, T 6 < +' 1,031bp 615bp,, " 0.012 ~ 0.006ng N 93 4 A, \ 6 8 6, DNA 9 P 9 = T6V, Z R ) < ', H I ^ H PCR, ) 3.4 =8 6 > > 5 (SingleStrandConformationPoly morphism,sscp) SSCP D * 0,G, E PCR < = 6 _ G ) \ DNA +, D,\ A, F, -.^, H A + ) SSCP 4 _ W E,, PW *B * G D 5 SSCP> A G,, G. / 9 DNA,\RA, F > SSCP9/ 6 P,5 P W, T 6, ; Lin [13] 9ITS1 2 V ", P 9 0 46, T 6 N 3 4, SSCP!,, T 6, T 6 ' DNA 2,, T 6,D Z B, I^ 9, ) Alasaad [46] P K B, Y PCR F O ) (F PCR SSCP)9 >, 40,T6, ITS2 234, P! A, T 6,T6, R )!,,5 < ) (? +1.4bp),/> > P /, T 6 "7 ) 9 0,H,T6*! D 3.5? @ DNA < (microsatelitedna) $ DNA? )/W 6 D 6 * 0, 9 1~6 EF\?( 2),* H- 2, * H - 2 (shorttandemrepeats,str) H - 2 (simple repeatsequence,srs SSR) / 6 A, 0, P PCR 9 <$ DNA,_ W ] Q & ',X,%8 * *, 4, )% 6,U, $ DNA, O) " 2H-,+ F\, / 6 B +, ^ c, ; H-)C D, c 5 ), U P, D Dar [47],T6$ 2 7 8 ^ 4 P 56, 142,T6 E,4 FM1 FM2 FM3 FM434R ) <, O PCR RFLP,,T6/ FM1 T +,,T6/ FM2 T +, D,3 *, O), * FM1 FM2 PW,T6, A 3.6 A B / 0 > 5 (Sequence relatedamplifiedpoly morphism,srap ) _ 1 V 6 Li Quiros [48] W2001 2 < O ), * +0 GC c c,h 2+ +0 AT c c, R 7856 * DNA (openreadingframe) 34 PCR <,*> B 2+ +,+'+ H = O) SRAP V, + / \ ] PW> 2G6,> U,,5 Y / 6,T6, G 0 P;> Li [49] P SRAP 9 10 5 6 0 5 P W <0 > > BT, 120,T6 E, 56 < O ) ' 15, X / 100~2000 bpd SRAP! A,T6,T6, R ) ', E ', C,T6 \ 0 ) E 9 X * 9 /(UPGMA)34,, T 6 *, bootstrap, A ; 3L c6 3.7 DNA 2 + 6 9 P W 6 A\,Z W DNA 2, + DNA 2,D W DNA F, /,% A 6,, 6 V 3LV /W &',GL,, c%,> C ^ = + 9, ), U PW 6, + Robert [50] >,T6,ITS2 34D, J B, T 6. G,4,T6, +?0.4% H,T6,T6 D 6 >, + 2.8% + +,R) [ITS2 V,T6 A, 0; -E,T 6ITS2 *, T 6 ),? +,* + W - E, T 6 Z W, T 6 Itagaki [51] ^ - E B, T 6, J B *
369 NAD1 COX1 2 C B D Z W, T 6,5 6 C ITS2 *,D \$, Z,7 -E B,T60 6,T6,? 1,T6 [17] 9 Z A> - E, T 6 Z W,T6,T6,, ) Amer [8] PITS1 ITS2 COX1 NAD14 + > BT,91,T634D, G 8'>, 0ITS1 6 G,ITS2 5 G,, T 6 / G H / JB *0,NAD1 68 G, 14,T6 * 19,T6 *,COX1 54 G, 13, T 6 * 7, T 6 * $',,,/ITS1 ITS20 A,T 6, N/ NAD1 COX1 0 Z W, T 6,T6, >, BT U D / P mtdna rdna,mtdna,l,? mtdna. /, T6 HrDNA a -,9 * H - 2,, Q ) /, B0,rDNA D TH [52], B> * 4 =+ +, 4, * / P, +9( PCR RFLP RAPD )0D 4, *, 0 T + W - 2, - *,* - * pepck pold PW,T6, [36] 4 0,,O /, T 6, A E A ^,_,,T6,T6,,O X +, " \ 6 6 B + R, 0, B 5, O A " A c, A_, ) B, O > ),, T 6,,?,OR./,T6, & :, +69 PW 3L, 6, ^, U P, + / RFLP SSCP RAPD ) / > 8 ', L, V ^ \E,,/ P0; A DNA 2 D / 9 * \6, +, &% A,G,9 Z `, P W A,O @, 6) U, DNA 2, \ + 9, (,U P, " rdna, ITS1 ITS2 mtdna, COX1 NAD1,,! 2 @ > 2 > \ 6 A,U,,< +, c / A8' 6,% ) +69, : 3^,T6,, >,T6, G +6R A6,BT ) 3 L R,, T 6, A ( `,- WX! [1] KeiserJ,UtzingerJ.Food bornetrematodiases[j].clinmicrobi olrev,2009,22(3):466-83. [2] MohammadAlizadeh AH,RoshaniM,LahmiF,etal.Cholan giocarcinomainmagneticresonancecholangiopancreatographyand fascioliasisinendoscopicultrasonography[j].caserepgastroen terol,2011,5(3):569-77. [3] Mas ComaS,ValeroM A,BarguesM D.Chapter2fasciola, lymnaeidsandhumanfascioliasis,withaglobaloverviewondis easetransmission,epidemiology,evolutionarygenetics,molecu larepidemiologyandcontrol[j].advparasitol,2009,69(chap ter2):41-146. [4] -,,., T 6 M, [J].0 KL ",2006,42(2):12-14. [5] YadavSC,SharmaR L,Kalicharan A,etal.Primaryexperi mentalinfectionofriverinebufaloeswithfasciolagigantica[j]. VetParasitol,1999,82(4):285. [6] LongC,DaugschiesA,WangBY,etal.Bloodeicosanoidsand immuneindicesduringfasciolosisinwaterbufaloes[j].parasitol Int,2000,49(4):273-8. [7] Robinson M W,TrudgetA,HoeyE M,etal.Triclabendazole resistant 犉犪狊犮犻狅犾犪犺犲狆犪狋犻犮犪 :beta tubulinandresponsetoinvitro treatmentwithtriclabendazole[j].parasitology,2002,124(pt 3):325-38. [8] AmerS,DarY,IchikawaM,etal.Identificationoffasciolaspe ciesisolatedfrom Egyptbasedonsequenceanalysisofgenomic (ITS1andITS2)andmitochondrial(NDIandCOI)genemarkers [J].ParasitolInt,2011,60(1):5-12. [9] LiuG H,GasserRB,YoungND,etal.Completemitochondrial genomesofthe'intermediateform'offasciolaand 犉犪狊犮犻狅犾犪犵犻犵犪狀 狋犻犮犪,andtheircomparisonwith 犉. 犺犲狆犪狋犻犮犪 [J].ParasitVectors, 2014,7(1):150. [10] AfshanK,ValeroM A,Qayyum M,etal.Phenotypesofinter mediateformsof 犉犪狊犮犻狅犾犪犺犲狆犪狋犻犮犪 and 犉. 犵犻犵犪狀狋犻犮犪 inbufaloes from CentralPunjab,Pakistan[J].JHelminthol,2014,88(4): 417-26. [11] IchikawaM,IwataN,ItagakiT.DNAtypesofaspermicFasci olaspeciesinjapan[j].jvetmedsci,2010,72(10):1371-4. [12] PeriagoM V,ValeroM A,SayedM E,etal.Firstphenotypic descriptionof 犉犪狊犮犻狅犾犪犵犻犵犪狀狋犻犮犪 intermediateformsfromthehu manendemicareaoftheniledelta,egypt[j].infectgenetevol,2008,8(1):51-58. [13] LinR Q,DongSJ,NieK,etal.Sequenceanalysisofthefirst internaltranscribedspacerofrdnasupportstheexistenceofthe intermediatefasciolabetween 犉. 犺犲狆犪狋犻犮犪 and 犉. 犵犻犵犪狀狋犻犮犪 in mainlandchina[j].parasitolres,2007,101(3):813-7. [14] AshrafiK,Valero M A,Panova M,etal.Phenotypicanalysis ofadultsof 犉犪狊犮犻狅犾犪犺犲狆犪狋犻犮犪, 犉犪狊犮犻狅犾犪犵犻犵犪狀狋犻犮犪 andintermedi ateformsfromtheendemicregionofgilan,iran[j].parasitol Int,2006,55(4):249-60. [15] FletcherHL,HoeyE M,OrrN,etal.Theoccurrenceandsig nificanceoftriploidyintheliverfluke, 犉犪狊犮犻狅犾犪犺犲狆犪狋犻犮犪 [J]. Parasitology,2004,128(Pt1):69. [16] SakaguchiY.Karyotypeandgametogenesisofthecommonliver fluke, 犉犪狊犮犻狅犾犪狊狆,inJapan[J].JapanJParasitol,1980,29(6): 507-13. [17] ItagakiT,TsutsumiK.TriploidformofFasciolainJapan:ge neticrelationshipsbetween 犉犪狊犮犻狅犾犪犺犲狆犪狋犻犮犪 and 犉犪狊犮犻狅犾犪犵犻犵犪狀 狋犻犮犪 determinedbyits 2sequenceofnuclearrDNA[J].IntJ Parasitol,1998,28(5):777. [18],, W. T 6 S B, [J].0 K L,1988,4(6):13-18. [19],T2.,T6, [J]. L,1988,8(1):53-6. [20] a # "., T 6 [J]. a ( L ", 1995,1(1):24-5. [21] a#".,t6 SB, [J]. G! ( 6),1988,4(2):117-8. [22] AgatsumaT,TerasakiK,YangL,etal.Geneticvariationin thetriploidsofjapanesefasciolaspecies,andrelationshipswith
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