投喂蚕豆对草鱼免疫器官超微结构和免疫基因表达的影响
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摘要
为研究草鱼摄食蚕豆后免疫功能变化及转投30 d人工饲料后免疫增强效果,在草鱼摄食蚕豆30、60、90、120及150 d(转投饲料30 d)时,检测肠道、头肾、肝脏等器官超微结构及C-type lysozyme、IFN-I、TNF-α等免疫基因表达变化。结果表明:1)饲喂单一饲料蚕豆的草鱼,其肠道、头肾、肝脏等3个重要免疫器官受到严重损伤,转投饲料30 d后其病变并未显著好转;2)免疫基因C-type lysozyme、IFN-I、TNF-α、IgM和MHC-I的表达在30～120 d均被抑制,转投饲料30 d后,头肾中C-type lysozyme、IL-1β、IFN-I、IgM、MHC-I以及肝脏中IL-1β、TNF-α、IFN-I、IgM、MHC-I的表达量有所增加。综上,草鱼摄食蚕豆后其免疫功能受到显著抑制,而转投30 d饲料对脆肉鲩的免疫机能具有一定的恢复作用。
In order to study the immune function changes of grass carp after feeding with broad bean and switching to artificial feed,immune organs including intestinal tract,head kidney and liver,and immune genes like C-type lysozyme,IFN-I,TNF-α,etc.were detected at for 30,60,90 and 120 and 150 d of feeding with broad bean(feeding with artificial feed for 30 d after feeding with broad bean for 120 d) in this study.The results showed that,the intestinal tract,head kidney and liver of tested grass carp were severely damaged after feeding with broad bean,and the damaged ultrastructure were not significantly improved after switching to artificial feed.The expression of immune genes C-type lysozyme,IFN-I TNF-α,IgM and MHC-I were all inhibited,and the expression of C-type lysozyme,IL-1β,IFN-I,IgM,MHC-I in the head kidney and IL-1β,TNF-α,IFN-I,IgM,and MHC-I in the liver increased after feeding with artificial feed.In summary,the immune function of grass carp was significantly inhibited after feeding with broad bean,and damaged immune function of crisp grass carp were slightly improved after feeding with artificial feed.This study would provide theoretical basis for the immune improvement of crisp grass carp and reference to investigate the impact of broad bean on the immune function of other animals.
In order to study the immune function changes of grass carp after feeding with broad bean and switching to artificial feed,immune organs including intestinal tract,head kidney and liver,and immune genes like C-type lysozyme,IFN-I,TNF-α,etc.were detected at for 30,60,90 and 120 and 150 d of feeding with broad bean(feeding with artificial feed for 30 d after feeding with broad bean for 120 d) in this study.The results showed that,the intestinal tract,head kidney and liver of tested grass carp were severely damaged after feeding with broad bean,and the damaged ultrastructure were not significantly improved after switching to artificial feed.The expression of immune genes C-type lysozyme,IFN-I TNF-α,IgM and MHC-I were all inhibited,and the expression of C-type lysozyme,IL-1β,IFN-I,IgM,MHC-I in the head kidney and IL-1β,TNF-α,IFN-I,IgM,and MHC-I in the liver increased after feeding with artificial feed.In summary,the immune function of grass carp was significantly inhibited after feeding with broad bean,and damaged immune function of crisp grass carp were slightly improved after feeding with artificial feed.This study would provide theoretical basis for the immune improvement of crisp grass carp and reference to investigate the impact of broad bean on the immune function of other animals.
引文
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[3] 谭乾开,黎华寿.食物与水环境因子对草鱼 (Ctenopharyngodon idellus C et Ⅴ) 脆化过程的影响[J].生态学报,2006,26(7):2409-2415Tan Q K,Li H S.Effects of diets dose and water quality on the transformation of common grass carp (Ctenopharyngodon idellus C et Ⅴ) to crisped grass carp[J].Acta Ecologica Sinica,2006,26(7):2409-2415 (in Chinese)
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[7] Martínez J A,Macarulla M T,Marcos R,Larralde J.Nutritional outcome and immunocompetence in micefed on a diet containing raw field beans (Vicia faba,var minor) as the source of protein[J].BritishJournal of Nutrition,1992,68(2):493-503
[8] De C S,Crampton V O,Bicskei B,Tocher D R.Replacement of dietary soy- with air classified faba bean protein concentrate alters the hepatic transcriptome in atlantic salmon (Salmo salar) parr[J].Comp Biochem Physiol Part D Genomics Proteomics,2015,16(3),48-58
[9] Gan L,Li X ?,Pan Q,Wu S ?,Feng T,Ye H,Gan L,Li X ?,Pan Q.Effects of replacing soybean meal with faba bean meal on growth,feed utilization and antioxidan tstatus of juvenile grass carp,Ctenopharyngodon idella[J].Aquaculture Nutrition,2017,23(1):192-200
[10] Partanen K,Alaviuhkola T,Siljander-Rasi H,Suomi K.Faba beans in diets for growing-finishing pigs[J].Agricultural & Food Science in Finland,2003,12(1):35-47
[11] Marquardt R R,Campbell L D.Performance of chicks fed faba bean (vicia faba) diets supplemented with methionine,sulfate and cystine[J].Canadian Journal of Animal Science,1975,55(2):213-218
[12] Ouraji H,Zaretabar A,Rahmani H.Performance of rainbow trout (Oncorhynchus mykiss) fingerlings fed diets containing different levels of faba bean (Vicia faba) meal[J].Aquaculture,2013,416-417:161-165
[13] McMillan D N,Secombes C J.Isolation of rainbow trout (Oncorhynchus mykiss) intestinal intraepithelial lymphocytes (IEL) and measurement of their cytotoxic activity[J].Fish & Shellfish Immunology,1997,7:527-541
[14] 卢全章.草鱼头肾免疫细胞超微结构的观察[J].水生生物学报,1997,21(2):152-156Lu Q Z.An ultrastructural observation of immune cells in pronephros of grass carp,Ctenopharyngodon idellus[J].Acta Hydrobiologica Sinica,1997,21(2):152-156 (in Chinese)
[15] 张振男,郁二蒙,谢骏,王广军,余德光,李志斐,龚望宝,王海英,魏南,夏耘.不同脆化阶段草鱼肠道菌群动态变化、血清酶指标及生长性能[J].农业生物技术学报,2015,23(2):151-160Zhang Z N,Yu E M,Xie J,Wang G J,Yu D G,Li Z F,Gong W B,Wang H Y,Wei N,Xia Y.Intestinal microflora dynamic change,serum enzyme and growth performance of the grass carp (Ctenopharyngodon idellus) at different stages of feeding broad bean (Vicia faba)[J].Journal of Agricultural Biotechnology,2015,23(2):151-160 (in Chinese)
[16] de Santis C D,Taylor J F,Martinez-Rubio L,Boltana S,Tocher D R.Influence of development and dietary phospholipid content and composition on intestinal transcriptome of Atlantic salmon (Salmo salar)[J].Plos One,2015,10(10):e0140964
[17] Esparza M L,Larralde J,Martínez J A.Development-related nutritional utilization and immune function in mice fed on two different protein sources[J].Nutrition Research,1996,16(5):829-839
[18] 罗华.警惕蚕豆病丧命[J].农村百事通,2015(13):62-63Luo F.Be wary of the death of Favism[J].Nongcun Baishitong,2015(13):62-63 (in Chinese)
[19] Kanda Y,Hisayasu S,Abe Y,Katsura K,Mashimo K.Growth-active peptides are produced from α-lactalbumin and lysozyme[J].Life Sciences,2007,81(6):449-457
[20] 陈度煌.投喂蚕豆对罗非鱼肝损伤及非特异性免疫功能的影响[J].中国饲料,2015,(13):24-26Chen D H.Effects of feeding broad bean on liver injury and non-specific immune function of tilapia[J].China Feed,2015,(13):24-26 (in Chinese)
[21] Dinarello C A.Interleukin-1[J].Cytokine and Growth Factor Reviews,1997,8(4):253-265
[22] Teles M,Mackenzie S,Boltaňa S,Callol A,Tort L.Gene expression and TNF-alpha secretion profile in rainbow trout macrophages following exposures to copper and bacterial lipopolysaccharide[J].Fish & Shellfish Immunology,2011,30(1):340-346
[23] Robertsen B.The interferon system of teleost fish[J].Fish & Shellfish Immunology,2006,20(2):172-191
[24] Peng L,Yang C,Su J.Protective roles of grass carp Ctenopharyngodon idella Mx isoforms against grass carp reovirus[J].PloS One,2012,7(12):e52142
[25] Scapigliati G,Buonocore F,Bird S,Zou J,Pelegrin P,Falasca C D,Prugnoli C J,Secombes C J.Phylogeny of cytokines:molecular cloning and expression analysis of sea bass Dicentrarchus labrax interleukin-1β[J].Fish & Shellfish Immunology,2001,11(8):711-726
[26] Liew F Y.The role of innate cytokines in inflammatory response[J].Immunology Letters,2003,85(2):131-134
[27] Wilson M,Bengten E,Miller N W,Clem L W,du Pasquier L,Warr G W.A novel chimeric Ig heavy chain from a teleost fish shares similarities to IgD[J].Proceedings of the National Academy of Sciences,1997,94(9):4593-4597
[28] Marzo P D F,Tosar P D A,Santidrian P D S.Influence of feeding a raw kidney bean diet (Phaseolus vulgaris L) on the immune function of growing chickens[J].Nutrition Research,1991,11(2-3):239-250
[29] Hou Y,Han X,Yuzuru S,Katumi A.Immune function of immature rainbow trout (Oncorhynchus mykiss) related with photoperiod[J].Zoological Research,1999,20(6):401-405
[30] Park K C,Osborne J A,Tsoi S C,Brown L L,Johnson S C.Expressed sequence tags analysis of Atlantic halibut (Hippoglossus) liver,kidney and spleen tissues following vaccination against Vibrio anguillarum and Aeromonas salmonicida[J].Fish & Shellfish Immunology,2005,18(5):393-415
[31] J?rgensen S M,Syvertsen B L,Lyngsyvertsen B,Lukacs M,Grimholt U,Gj?en T.Expression of MHC class I pathway genes in response to infectious salmon anaemia virus in Atlantic salmon (Salmo salar L) cells[J].Fish & Shellfish Immunology,2006,21(5):548-560
[2] 麦康森.转变增长方式是我国水产养殖持续发展的必由之路[J].渔业信息与战略,2012,27(1):1-6Mai K S.Change of growth pattern and sustainable development of aquaculture in China[J].Fisheries Information & Strategy,2012,27(1):1-6 (in Chinese)
[3] 谭乾开,黎华寿.食物与水环境因子对草鱼 (Ctenopharyngodon idellus C et Ⅴ) 脆化过程的影响[J].生态学报,2006,26(7):2409-2415Tan Q K,Li H S.Effects of diets dose and water quality on the transformation of common grass carp (Ctenopharyngodon idellus C et Ⅴ) to crisped grass carp[J].Acta Ecologica Sinica,2006,26(7):2409-2415 (in Chinese)
[4] 郁二蒙,谢骏,卢炳国,王广军,余德光.脆肉鲩与普通草鱼肌肉显微结构观察[J].南方农业学报,2014(4):671-675Yu E M,Xie J,Lu B G,Wang G J,Yu D G.Microstructure of muscles between crisp grass carp and common grass carp[J].Journal of Southern Agriculture,2014(4):671-675 (in Chinese)
[5] 刘怡.《海洋与渔业》走进“中国脆肉鲩之乡”中山东升镇[J].海洋与渔业,2014(3):30-33Liu Y.Ocean and Fishery entered the “Hometown of Chinese crispy meat” in Zhongshan,Dongsheng Town[J].Ocean and Fishery,2014,(3):30-33 (in Chinese)
[6] 吴康,黄晓声,金洁南,李男,张学振.饲喂蚕豆对草鱼抗氧化能力及免疫机能的影响[J].水生生物学报,2015(2):250-258Wu K,Huang X S,Jin J N,Li N,Zhang X Z.Effects of feeding with faba bean (Vicia faba) on the antioxidant capability and immune functions of grass carp (Ctenopharyngodon idellu) [J].Acta Hydrobiologica Sinica,2015(2):250-258 (in Chinese)
[7] Martínez J A,Macarulla M T,Marcos R,Larralde J.Nutritional outcome and immunocompetence in micefed on a diet containing raw field beans (Vicia faba,var minor) as the source of protein[J].BritishJournal of Nutrition,1992,68(2):493-503
[8] De C S,Crampton V O,Bicskei B,Tocher D R.Replacement of dietary soy- with air classified faba bean protein concentrate alters the hepatic transcriptome in atlantic salmon (Salmo salar) parr[J].Comp Biochem Physiol Part D Genomics Proteomics,2015,16(3),48-58
[9] Gan L,Li X ?,Pan Q,Wu S ?,Feng T,Ye H,Gan L,Li X ?,Pan Q.Effects of replacing soybean meal with faba bean meal on growth,feed utilization and antioxidan tstatus of juvenile grass carp,Ctenopharyngodon idella[J].Aquaculture Nutrition,2017,23(1):192-200
[10] Partanen K,Alaviuhkola T,Siljander-Rasi H,Suomi K.Faba beans in diets for growing-finishing pigs[J].Agricultural & Food Science in Finland,2003,12(1):35-47
[11] Marquardt R R,Campbell L D.Performance of chicks fed faba bean (vicia faba) diets supplemented with methionine,sulfate and cystine[J].Canadian Journal of Animal Science,1975,55(2):213-218
[12] Ouraji H,Zaretabar A,Rahmani H.Performance of rainbow trout (Oncorhynchus mykiss) fingerlings fed diets containing different levels of faba bean (Vicia faba) meal[J].Aquaculture,2013,416-417:161-165
[13] McMillan D N,Secombes C J.Isolation of rainbow trout (Oncorhynchus mykiss) intestinal intraepithelial lymphocytes (IEL) and measurement of their cytotoxic activity[J].Fish & Shellfish Immunology,1997,7:527-541
[14] 卢全章.草鱼头肾免疫细胞超微结构的观察[J].水生生物学报,1997,21(2):152-156Lu Q Z.An ultrastructural observation of immune cells in pronephros of grass carp,Ctenopharyngodon idellus[J].Acta Hydrobiologica Sinica,1997,21(2):152-156 (in Chinese)
[15] 张振男,郁二蒙,谢骏,王广军,余德光,李志斐,龚望宝,王海英,魏南,夏耘.不同脆化阶段草鱼肠道菌群动态变化、血清酶指标及生长性能[J].农业生物技术学报,2015,23(2):151-160Zhang Z N,Yu E M,Xie J,Wang G J,Yu D G,Li Z F,Gong W B,Wang H Y,Wei N,Xia Y.Intestinal microflora dynamic change,serum enzyme and growth performance of the grass carp (Ctenopharyngodon idellus) at different stages of feeding broad bean (Vicia faba)[J].Journal of Agricultural Biotechnology,2015,23(2):151-160 (in Chinese)
[16] de Santis C D,Taylor J F,Martinez-Rubio L,Boltana S,Tocher D R.Influence of development and dietary phospholipid content and composition on intestinal transcriptome of Atlantic salmon (Salmo salar)[J].Plos One,2015,10(10):e0140964
[17] Esparza M L,Larralde J,Martínez J A.Development-related nutritional utilization and immune function in mice fed on two different protein sources[J].Nutrition Research,1996,16(5):829-839
[18] 罗华.警惕蚕豆病丧命[J].农村百事通,2015(13):62-63Luo F.Be wary of the death of Favism[J].Nongcun Baishitong,2015(13):62-63 (in Chinese)
[19] Kanda Y,Hisayasu S,Abe Y,Katsura K,Mashimo K.Growth-active peptides are produced from α-lactalbumin and lysozyme[J].Life Sciences,2007,81(6):449-457
[20] 陈度煌.投喂蚕豆对罗非鱼肝损伤及非特异性免疫功能的影响[J].中国饲料,2015,(13):24-26Chen D H.Effects of feeding broad bean on liver injury and non-specific immune function of tilapia[J].China Feed,2015,(13):24-26 (in Chinese)
[21] Dinarello C A.Interleukin-1[J].Cytokine and Growth Factor Reviews,1997,8(4):253-265
[22] Teles M,Mackenzie S,Boltaňa S,Callol A,Tort L.Gene expression and TNF-alpha secretion profile in rainbow trout macrophages following exposures to copper and bacterial lipopolysaccharide[J].Fish & Shellfish Immunology,2011,30(1):340-346
[23] Robertsen B.The interferon system of teleost fish[J].Fish & Shellfish Immunology,2006,20(2):172-191
[24] Peng L,Yang C,Su J.Protective roles of grass carp Ctenopharyngodon idella Mx isoforms against grass carp reovirus[J].PloS One,2012,7(12):e52142
[25] Scapigliati G,Buonocore F,Bird S,Zou J,Pelegrin P,Falasca C D,Prugnoli C J,Secombes C J.Phylogeny of cytokines:molecular cloning and expression analysis of sea bass Dicentrarchus labrax interleukin-1β[J].Fish & Shellfish Immunology,2001,11(8):711-726
[26] Liew F Y.The role of innate cytokines in inflammatory response[J].Immunology Letters,2003,85(2):131-134
[27] Wilson M,Bengten E,Miller N W,Clem L W,du Pasquier L,Warr G W.A novel chimeric Ig heavy chain from a teleost fish shares similarities to IgD[J].Proceedings of the National Academy of Sciences,1997,94(9):4593-4597
[28] Marzo P D F,Tosar P D A,Santidrian P D S.Influence of feeding a raw kidney bean diet (Phaseolus vulgaris L) on the immune function of growing chickens[J].Nutrition Research,1991,11(2-3):239-250
[29] Hou Y,Han X,Yuzuru S,Katumi A.Immune function of immature rainbow trout (Oncorhynchus mykiss) related with photoperiod[J].Zoological Research,1999,20(6):401-405
[30] Park K C,Osborne J A,Tsoi S C,Brown L L,Johnson S C.Expressed sequence tags analysis of Atlantic halibut (Hippoglossus) liver,kidney and spleen tissues following vaccination against Vibrio anguillarum and Aeromonas salmonicida[J].Fish & Shellfish Immunology,2005,18(5):393-415
[31] J?rgensen S M,Syvertsen B L,Lyngsyvertsen B,Lukacs M,Grimholt U,Gj?en T.Expression of MHC class I pathway genes in response to infectious salmon anaemia virus in Atlantic salmon (Salmo salar L) cells[J].Fish & Shellfish Immunology,2006,21(5):548-560