; E lm an - ; Fernando [15 ], TCG, Jour. of N o rthw est Sci2T ech U niv. of A gri. and Fo r. (N aṫ Sci. Ed. ) Jun.

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1 34 6 () V o l. 34 N o Jour. of N o rthw est Sci2T ech U niv. of A gri. and Fo r. (N aṫ Sci. Ed. ) Jun Ξ 1, 1, 2, 1, 1, 1, 1, 1 (1, ; 2, ) [ ], 3, ggml,, ( ) %, ( ) % ( ) %, (P > 0. 05), (P < 0. 05); ( ) %, ( ) %, (P < 0. 05), 3 ; ( ) % ( ) %, (P < 0. 05), ggml [ ] ; ; ; ; [] S [] A [ ] (2006) Po lge [1 ],,,, -, ;, [223 ],, [4 ],,,,,,,, [528 ],, Bayard [9 ],D al2 im ata [10 ] W oelders [11 ] ; E lm an [12 ] E im an [13 ] - ; [14 ] - ; Fernando [15 ], TCG, 3, -,, Ξ [ ] [ ] (2004K022G11203, 2005K022G03203) ; (05ZR 100) [ ] (1976- ),,,, [ ] (1956- ),,,,, E2m ail: liqingw com. cn

2 42 () 34 8,, 1 1, : D 2, (F ITC2PNA ) (P I) CTC 3 Sigm a ; ; : B IO TRON 2 ICS ; Barnan t ; ; PH S23C ph ; O SM OM A T 030 ; ( 1) TCG [16 ] 1. 1 g, g, T ris g, 100 ml, ph 6. 21, 286 mo sm ( 2) BT S 3. 7 g, ED TA g, 0. 6 g, N ahco g, KC l g, g, 0. 1 g, 100 ml, ph 7. 25, 315 mo sm (3) P I( ) g P I 2 ml PBS, Λm, - 20 (4) g, g, 50 ml, 150 mo sm ( 5) g ml (DM SO ), Λm, (6)CTC T ris g, N ac l g, L g, CTC g, 10 ml, ph 7. 8, Λm, (7) (F ITC2 PNA ) g F ITC2PNA, 10 ml PBS, Λm, mmo lgl, TCG, , , 0105, 0140 ggml, TCG 3,, P I2 123 F ITC2PNA CTC 3, (HO ST ) 1. 3, 50 ml, (800 g, 10 m in), BT S, h, (800 g, 10 m in),, g (TCG + 20% ), 5 2 h, g (3% g ), 5 1 h, (0. 25 ml ), 1 gm in 5-5, 3 cm 15 m in, , 37, 30 s, 10 ΛL,, 400 ( ) BT S, gml 100 ΛL H EPESgBSA [16 ] 1. 5 ml, 10 ΛL P I 1 ΛL 123, 10 m in, 50 ΛL, m in 10 ΛL,, (400 ) 488 nm,, ( ) ( ) (HO ST ), gml, m in, 20 ΛL, 400,, L am ia [17 ], F ITC2PNA, PV P,

3 6 : g 6 m in, ; PBS, gml, 30 ΛL,, 10 m in, 30 ΛL F ITC2PNA, m in, PBS, (10% PBS ),, BT S, gml, 400 ΛL 1 ml 30 ggl PV P, 500 g 6 m in,, (1) 200 ΛL PBS, 200 ΛL CTC, 30 s, 36 ΛL 12. 5%, 10 ΛL,,, 400 (2) 200 ΛL TCM, 3 h, 200, 1. 5 SPSS O ne2w ay ANOVA ,, ggml, (P < 0. 05) ;, ;, ( ) % ( ) %, TCG (n= 21) T able 1 V iability,mo tility and the percentages of m itochondrial activity sperm after frozen2thaw ed w ith disaccharides(n= 21) Group g (g ml - 1 ) Concentration g% V iability g% M o tility M itochondrial activity TCG Contro l a a a T rehalo se b b b a a a c c c d c d Sucro se b b e e a a cd c c d c d L acto se e a a f a a cd c c d c d : (P < 0. 05) N o te: V alues w ith different sm all letters w ith in the sam e co lum n differ significantly. Sam e in the fo llow ing tables , ggml,, TCG (P < 0. 05),, (P > 0. 05) (P > 0. 05), , ggml,, ( ) %, ( ) % ( ) %, (P < 0. 05),

4 44 () 34 TCG (P < 0. 05) , ggml, Group (P < 0. 05), ( ) % ( ) %, ( P < 0. 05) ; 3, (n= 21) T able 2 Effect of 3 disaccharides on the param eters of m em brane integrity, acro som e integrity g (g ml - 1 ) Concentration and capacitation of frozen2thaw ed sperm M em brane integrity A cro som e integrity Rate of capacitation sperm Befo re capacitation A fter capacitation TCG Contro l a ac a a T rehalo se b b b b c ) b b a d b b c de b b c Sucro se b ac a c a ac a d d a c c d a c e L acto se a c a d c c c a d ac c c e ac c e ,, ggml, ( ) %, (P < 0. 05) ;,, (P > 0. 05) ,,, - (P < 0. 05) ;, 3 (P < 0. 05), ggml,, ( ) %, ( ) % ( ) % 3, ggml,,,, [15, ],,,, E lm an [12 ],, ggml,,, ;, E lm an [12 ],,,,,, [15 ],,

5 6 : 45,,, Garcia [19 ],, -,,,,,, Jelinkova [20 ],,,, [ ] [ 1 ] Po lge C. A rtifical insem ination in p ig[j ]. V et Pec, 1956, 68: [ 2 ] L isa M T hurston,w illiam V Ho lt, Paul F W astson. Po st2thaw functional status of boar sperm atozoa cryop reserved using th ree contro lled rate freezers: a comparison[j ]. Theriogeno logy, 2003, 60: [ 3 ] W oelders H, M atth ijs A, Zuidberg C A, et al. C ryop reservation of boar sem en: equilibrium freezing in the cryom icro scope and in straw s[j ]. Theriogeno logy, 2005, 63: [ 4 ] Kum ar,m illar,w aston. T he effect of coo ling rate on the survival of cryop reserved bull, ram, and boar sperm atoaoa: a comparison of two contro lled2rate coo ling m achines[j ]. C ryobio logy, 2003, 46: [ 5 ],. [J ]., 2003, 5 (1) : [ 6 ],. [J ]., 2001, 21 (3) : [ 7 ] A lex, H ans. P reservation m echanism s of trehalo se in food and bio system s[j ]. Co llo ids and Surfaces B: B io interfaces, 2002, 57: [ 8 ],. [J ]. :, 2004, 19 (6) : [ 9 ] Bayarad, E sther, Kath leen. Comparison of glycero l, o ther po lyo ls, trehalo se, and raffino se to p rovide a defined cryop ro tectant m edium fo r mouse sperm cryop reservation[j ]. C ryobio logy, 1998, 37: [ 10 ] D alim ata, Graham. C ryop reservation on rabbit sperm atozoa using acetam ide in com bination w ith trehalo se and m ethyl cellulo se[j ]. T he2 riogeno logy, 1997, 48: [ 11 ] W oelders,m atth ijs, Engel. Effects of trehalo se and sucro se, o smo lality of the freezing m edium, and coo ling rate on viability and intact2 ness of bull sperm after freezing and thaw ing[j ]. C ryobio logy, 1997, 35: [ 12 ] E lm an, A boagla, T erada. T rehalo se2enhanced fluidity of the goat sperm m em ebrane and its p ro tection during freezing[j ]. B io logy of Re2 p roduction, 2003, 69: [ 13 ] E im an, T akato. Effects of the supp lem entation of trehalo se extender containing egg yo lk w ith sodium dodecyl sulfate on the freezability of goat sperm atozoa[j ]. Theriogeno logy, 2004, 62: [ 14 ],,,. [J ]., 1997, 28 (2) : [ 15 ] Fernando, M argareta, Szabo lcs, et al. D eep freezing of concentrated boar sperm fo r intra2uterine insem ination: effects on sperm viability[j ]. Theriogeno logy, 2005, 63: [ 16 ] E rik sson, Rodriguesz2M artinez. Effect of freezing and thaw ing rates on the po st2thaw viability of boar sperm atozoa frozen in F lat Pack s and M axi2straw s[j ]. A nim al Rep reduction Science, 2000, 63: [ 17 ] L am ia, D aniel, L aetitia. Bull sem en in v itro fertility after cryop reservation using egg yo lk LDL : a comparison w ith Op tidyl, a comm ercial egg yo lk extender[j ]. Theriogeno logy, 2004, 61: [ 18 ] F riedler S, Giudice L C, L am b E J. C ryop reservation of em bryo s and ova[j ]. Fertil Steril, 1988, 49: [ 19 ] Garcia D e Castro A, T unnacliffe A. Intracellular trehalo se imp roves o smo2to lerance but no t desiccatio to lerancein m amm alian cells[j ]. FEBS L etters, 2000, 487: [ 20 ] Jelinkova L, Selm an H A, A rav A, et al. Tw in p regnancy after vitrification of 22p ronuclei hum an em bryo s[j ]. Fertil Steril, 2002, 76 (2) : ( 51 )

6 6 : PA 51 C lon ing and imm unop ro tection of PA recom b inan t p ro tein of B acillus an th racis X IE Y ing-guo 1, 2,W U X iu-fang 1, 2,W U Su-q in 2,W ANG X i- liang 2, HE W e i-m ing 1 (1 Colleg e of A nim al S cience and T echonology,n orthw est A & F U niversity, Y ang ling, S haanx i , Ch ina; 2 D ep arm ent of Imm unology of R eseach Institute of M icrobiology and Ep idem iology A cademy of M ilitary M ed ical S cience, S tate K ey L aboratory of P athogen and B iosecurity,b eijing , China) Abstract: PA gene amp lified w as from B acillus an th racis by PCR and then cloned in to pet 28a (+ ) vecto r. T he recom b inan t p lasm id pet 28a (+ ) 2PA w as con structed and exp ressed and the imm unogen icity and imm unop ro tection w ere studied. T he resu lts show that the length of PA gene is abou t bp and rel2 ative mo lecu lar m ass of the exp ressed p ro tein is 83 ku; the rpa has good imm unogen icity, and specificity of an tibody cou ld be detected after 10 d imm un ization, and the an tibody levels rise m arkedly, 3rd to ; the rpa p ro tein stim u lates the specificity of an tibody b inding to mou se IgG12based; the rpa p ro tein can p romo te the grow th and m u ltip lication of the mou se sp leen cells in v itro. T he rpa imm un ization group can sign ifican tly increase CD + 4 mou se sp leen cells and the imm une respon ses tend to T h22type respon se, show ing that the rpa mo st m ediate to humo ral imm un ity. T he rpa is fairly imm une p ro tective, the p ro tec2 tion rate being 50%. Key words: B acillus an th racis; p ro tective an tigen; imm un ity; genetic engineering vaccine ( 45 ) Abstract ID: (2006) EA Effect of trehalo se, sucro se and lacto seon on cryop reservation of boar sperm ZHANG Shu- shan 1,L I Qing-wang 1, 2,L I Gang 1, CHEN X iao-yu 1, HU J ian-hong 1, HAN Zeng- sheng 1,W ANG L i-q iang 1 (1 Colleg e of A nim al S cience and T echnology, N orthw est A & F U niversity, Y ang ling, S haanx i , Ch ina; 2 D ep artm ent of B iolog ical Eng ineering, Collage of Env ironm ent & Chem ical Eng ineering, Yanshan U niversity, Q inghuangdao, H ebei , China) Abstract: In th is study, the effects of adding th ree disaccharides (trehalo se, sucro se and lacto se) in the cryop reservative so lu tion w ere compared. T he resu lts w ere summ arized as fo llow s: T he effect of boar se2 m en cryop reservation w as m arkedly imp roved w ith th ree disaccharides, and their op tim um level w as ggml ; A t the level of ggml, the viab ility, mo tility, the percen tages of m itochondrial activity and m em b rane in tegrity of frozen2thaw ed sperm w ith trehalo se w ere ( ) %, ( ) %, ( ) % and ( ) % respectively, and there w as no sign ifican t difference betw een tre2 halo se and sucro se (P > 0. 05), and bo th of them w ere h igher than that of lacto se group and the con tro l (P < 0. 05) ; T he percen tage of in tact acro som e of trehalo se group w as ( ) %, and h igher than that of the sucro se, lacto se group, and the con tro l (P < 0. 05), and increased w ith the increasing level of th ree disaccharides; T he percen tages of capacitating boar sperm befo re and after capacitation treatm en t w ere ( ) % and ( ) %, and w as better than that of lacto se group and the con tro l(p < 0. 05). To sum up, the effect of imp roving the frozen2thaw ed sperm quality w ith trehalo se w as superio r to sucro se, lacto se and TCG con tro l, and the op tim um level w as ggml. Key words: boar sem en; cryop reservation; trehalo se; sucro se; lacto se