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1 49 (6) : , 2003 A cta Zoologica S inica C 3 C3 C4 33 (, ) (, ) C ( Pelodiscus sinensis) C3 C4, 6, C mg/ kg, 4, C3 C4, C 250 mg/ kg, C3 ; C mg/ kg 4, C3 C 250 mg/ kg ; C 500 mg/ kg, C4 5 ; C 250 mg/ kg mg/ kg, C3, C mg/ kg, C mg/ kg 3, C4, C mg/ kg 3, C4 C C3 C4, C, C3 C4,, C [ 49 (6) : , 2003 ] C C3 C4 Effects of vitamin C and acid2stress on serum complement C3 and C4 level in juvenile soft2shelled turtles ( Pelodiscus sinensis) 3 ZHOU Xian2Qing N IU Cui2J uan 33 SUN Ru2Yong ( Mi nist ry of Education Key L aboratory f or Biodiversity Science and Ecological Engi neeri ng, Instit ute of L if e Sciences, Beiji ng Normal U niversity, Beiji ng , Chi na) ( Depart ment of L aboratory A ni mal Sciences, Capital U niversity of Medical Sciences, Beiji ng , Chi na) Abstract This study was conducted to investigated the effects of dietary vitamin C on serum complement C3 and C4 in stressed and unstressed juvenile soft2shelled turtle. The turtles were fed with vitamin C supplementation at dosages of 0, 250, 500, 2 500, and mg/ kg diets, respectively for 4 weeks. Blood was collected from turtles and levels of serum complement C3 and C4 analyzed by the transmission colorimetric method. Levels of serum complement C3 between the control group and the group supplemented vitamin C at 250 mg/ kg diet showed no significant difference. The levels of serum complement C3 in the four groups supplemented with vitamin C at 500, 2 500, and mg/ kg diet were significantly higher compared with the control group. The level of serum complement C4 in the group supplemented vita2 min C at 500 mg/ kg diet was distinctly higher than the other five groups, and in the group supplemented with vitamin C at 250 mg/ kg diet was higher than mg/ kg group. After acid stress, the complement C3 levels had no significant difference compared with the ones of before stress, while there was a significant decrease after stress when the three , [ This research was funded by a grant from Open Fund of State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences] 33 (Corresponding author). E2mail : edu. cn,, : : , E2mail : com. cn ν 2003 Acta Zoologica Sinica

2 groups supplemented vitamin C at 0, 250 and 500 mg/ kg diet were put together to analyze. The levels of serum comple2 ment C4 in the three groups supplemented with vitamin C at 0, 250 and 500 mg/ kg diet decreased significantly, while the other groups at 2 500, and mg/ kg diet had no significant difference after acid stress. No interaction effects of vitamin C and acid2stress on the levels of serum complement C3 and C4 were observed. These results suggest that vita2 min C seems to have an upper and lower threshold for improving levels of serum complement C3 and C4 in turtles, and acid stress decreases the their levels, while the high dosage vitamin C can prevent this decline [ Acta Zoologica Sinica 49 (6) : , 2003 ]. Key words Soft2shelled turtle, Vitamin C, Acid stress, Complement C3, Complement C4,,, C, C ( Penaeus chinensis), ( C, p H, 1996) ; ( O ncorhynchus mykiss ) C3 C4 (Verlhac et al., 1996) ; ( S parus aurata) (Ortuno et al., 2001) 111 ( Zhou et al., 2002) ;,, C 1, ( Sarna, et al., 1993) C 6 : 00 1, C, 2 h,,,, ,,, 3, (, 1997), C ( Hemi2 la, 1997), ( Dela2, (1998) fuente et al., 1986) C , g, 72 6,,, 12 2, 6 ( ),, 60 cm 30 cm 40 cm 6, C, C4 6 C, C (, 2000) mg/ kg C C ( C 90 %) (Lambris, 1989), - 20, 4 h ( Pelodiscus sinensis), 4,,,, 4 C, p H 719, p H 510, 24 h,

3 6 : C C3 C4 113 C3 C4 (, t, : t 1, 9 = t 1,8 = t 1,9 =, ) t 1,9 = t 1,9 = t 1,8 = 01751, min, P > 0105) C nm (A), 500 mg/ kg, C3 C4 Statistica C mg/ kg,, P < 0105, C, C C3, C 01398, P < 01848) C3 ( F 5,64 = 212 C , P < 0101) L SD, C 250 mg/ kg, C3 ( F 5,66 = 81079, P < 0101) L SD ( P > 0105) ; C, C 500 mg/ kg, mg/ kg 4, C3 C C 250 mg/ kg mg/ kg, C3, C C4 C ( 1), ( P < 0101), C mg/ kg, mg/ kg C C3 ( F 5,64 = C C4 C4 5 ( P < 0105) ; 250 mg/ kg ( P < 0105) ; C mg/ kg ( P < 0105) 2, C C3 ( ) Fig. 1 Effects of vitamin C on content of serum complement C3 in juvenile soft2shelled turtles ( Mean SD) LSD, LSD, [LSD multiple range test, different letters above the bars indicate [LSD multiple range test, different letters above the bars indicate significant difference among different dose groups ( P < 0105) ] significant difference among different dose groups ( P < 0105) ], ( F 1,66 = , P < 0101) t, C3 ( F 1,64 = 61875, P 2 C C4 ( ) Fig. 2 Effects of vitamin C on content of serum complement C4 in juvenile soft2shelled turtles ( Mean SD) C4, C mg/ < 0105) t, C3 kg, C4 ( t 1, 9 =

4 t 1,10 = t 1, 9 = 21305, P < 0105), C mg/ kg ( t 1,10 ( Porter et al., = t 1, 9 = t 1, 8 = 11888, P > 0105) ( 3), C mg/ kg, C3,, C3 ; C C4 C 500 mg/ kg, ( F 5, 66 = 01418, P < 01834) C3 3 C4 ( Pagrus pagrus) 2, ( ) ( Rotllant et al., Fig. 3 Effects of acid2stresse on content of serum complement C4 in juvenile soft2shelled turtles ( Mean SD) t, ( 3 ) [ t2test, the asterisks ( 3 ) above the bars indicate signifi2 cant difference per dose groups before and after acid2stress ( P < 0105) ] 3 (Johnston et al., 1987) C1q, 1978), C C4,, C, ; C4 C C3 312 C C3 C4, Sunyer et al. (1995), ( S parus aurata) 8 min, 16, 1997), mg/ kg, C3 ; C4 ( mg/ kg), C3 C4,,,,, C3 C4 ; 311 C C3 C4 C C3 C4,, C C C mg/ kg : (1) C mg/ kg,,, ( Verl2 (Umegaki et al., 1997), hac et al., 1996 ), C, mg/ kg, (Nunes et al., 1997) C, ( Verlhac et al., ( 1994) C mg/ kg, ), (Li et al.,, C 1985), Hardie et al. (1991),,,,, C 50 mg/ 100 g,, C1q,, C1q, (,

5 6 : C C3 C4 2000) C, tients. C, C ( Zhou et al., 2003) (2), C,,, 5 ( Zhou et al., 2003), (, 1996), C C3 C4 ;, ; C C3 C4, C,,, ( References) Delafuente, J. C., J. M. Prendergast and A. Modigh 1986 Im2 munolgic modulation by vitamin C in the elderly. Int. J. Im2 m unopharmacol. 8 (2) : Hardie, L. J., T. C. Fletcher and C. J. Secombes 1991 The ef2 fect of dietary vitamin C on the immune response of the Atlantic salmon ( S al mo salar L. ). A quacult ure 95 : Hemila, H Vitamin C supplementation and the common cold was linus pauling right or wrong? Internat. J. N ut r. Res. 67 : Hou, J. C., Y. D. Wu and Y. L. Ling 2000 Modulation of the in2 flammatory response through complement2neutrophil activation feedback mechanism with selenium and vitamin E. Acta Academ. Medic. Si ni. 22 (6) : [,, Wen, J The study progress of vitamin nutrition in Animals E - In : The Animal Nutrient Brabch of China Poultry Veterinary As2. 22 ( 6) : ] Johnston, C. S., W. P. Kolb and B. E. Haskell 1987 The effect of vitamin C nutriture on complement component C1q concentra2 tions in guinea pig plasma. J. N ut r. 117 (4) : Lambris, J. D The third component of the complement sys2 tem. Chemistry and biology. Curr. Top. Microbiol. Im m unol. 153 : Li, Y. and R. T. Loveell 1985 Elevated levels of dietary ascirbic acid increase immune responses in channel catfish. J. N ut r. 115 : Li, G. P., S. Y. Tao and B. Guo 2000 Changes in serum im2 munoglobulins, complements and C2reactive protein in trauma pa2 Chi nese Critical Care Medici ne 12 (2) : [,, 2000 C (2) : ] Nunes, G. L., K. Robinson, A. Kalynych, S. B. King, D. S. Sgoutas and B. C. Berk 1997 Vitamin C and E inhibit O 22 pro2 duction in the pig coronary artery. Ci rculation 96 (10) : Ortuno, J., A. Cuesta, M. Angeles2Esteban and J. Meseguer 2001 Effect of oral administration of high vitamin C and E dosages on the gilthead seabream ( S parus aurata L. ) innate immune system. Vet. Im m unol. Im m unopathol. 79 (3/ 4) : Porter, R. R. and K. B. M. Reid 1978 The biochemistry of com2 plement. N at ure 275 : Rotllant, J., M. Pavlidis, M. Kentouri, M. E. Abad and L. Tort 1997 Non2specific immune responses in the red porgy Pagrus pa2 grus after crowding stress. A quacult ure 156 : Sarna, S. and R. K. Bhola 1993 Chemo2immunotherapeutical stud2 ies on Dalton s lymphoma in mice using cisplatin and ascorbic acid : synergistic antitumor effect i n vivo and i n vit ro. A rch Im m unol. Ther. Ex p ( W arsz). 41 (5/ 6) : Sunyer, J. O., E. Gomez, V. Navarro, J. Quesada and L. Tort 1995 Physiological responses and depression of humoral compo2 nents of the immune system in gilthead sea bream ( S parus aurata) following daily acute stress. Can. J. Fish. A quat. Sci. 52 (11) : Umegaki, K., H. Uramoto and T. Esashi 1997 Lack of influence of a long2term high or low vitamin E diet on the oxidative DNA damage in the bone marrow of mice. Internat. J. V it. N ut r. Res. 67 : Verlhac, V. and J. Gabaudan 1994 Influence of vitamin C on the immune system of salmonids. A quat. Fish. Manag. 25 : Verlhac, V., J. Gabaudan, A. Obach, W. Schuep and R. Hole 1996 Influence of dietary glucan and vitamin C on non2specific and specific immune responses of rainbow trout mykiss). A quacult ure 143 : ( Oncorhynchus sociation ed. The Study Progress of Animal Nutrition. Beijing : China Agriculture University Press, [1997. :.. :, ] Wang, W. Q. and A. J. Li 1996 Influences of LAPP on the growth, resistance to low oxygen content and immunoresistance of Penaeus chi nensis. Transact. Oceanol. L i m nol. 1 : [, 1996 LAPP ( Penaeus chinen2 sis).. 1 : ] Yang, H. C Animal Immunology. Beijing : China Agricul2 ture University Press, [ 1996.

6 :, ] Zhou, X. Q., C. J. Niu and Q. F. Li 1998 The effects of light in2 tensity on daily food consumption and specific growth rate of the ju2 venile soft2shelled turtles, Trionyx sinensis. Acta Zool. Sin. 44 (2) : [,, (2) : ] Zhou, X. Q., C. J. Niu, R. Y. Sun and Q. F. Li 2002 The effect of Vitamin C on non2specific immune responses of the juve2 nile soft2shelled turtle ( Trionyx sinensis ). Comp. Biochem. Physiol. Part A, 131 (4) : Zhou, X. Q., M. X. Xie, C. J. Niu and R. Y. Sun 2003 The ef2 fects of dietary vitamin C on growth, liver vitamin C and serum cortisol levels in stressed and unstressed juvenile soft2shelled turtles ( Pelodiscus sinensis). Com par. Biochem. Physiol. Part A, 135 (2) :

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