鲈鲤仔稚鱼肠道形态发育及消化酶活性分析
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摘要
【目的】旨在揭示鲈鲤仔稚鱼肠道早期发育形态结构与功能的关系。【方法】通过形态观察和常规石蜡切片法观察了鲈鲤仔稚鱼肠道形态发育特征,并通过酶联免疫吸附法检测了其发育过程中胰蛋白酶、胰凝乳蛋白酶、α-淀粉酶和脂肪酶的比活力。【结果】在水温(20.6±0.7)℃条件下,初孵仔鱼肠道未分化,出膜后第5天(days after hatching,dah)肠道贯通,肠道"Z"形折点分别出现在30 dah和33 dah,此后形态变化不明显。伴随肠道形态变化,各肠段肠黏膜褶皱高度呈增高的趋势,且一直保持前肠>中肠>后肠。未分化肠道就能检测到胰蛋白酶、胰凝乳蛋白酶、α-淀粉酶和脂肪酶活性;肠道贯通过程中,胰蛋白酶和胰凝乳蛋白酶呈上升的趋势,α-淀粉酶变化不明显,而脂肪酶则呈下降的趋势。肠道出现"Z"形折点过程中,4种消化酶比活力显著增加;肠道形态稳定之后,4种消化酶比活力均呈下降趋势。【结论】实验室养殖条件下,鲈鲤仔稚鱼体长和体重生长分别呈线性和三项式关系;出膜后35 d,肠道形态发育基本稳定;消化酶的比活力随仔稚鱼肠道的发育变化显著。
【Objective】The objective of this study was to reveal the relationship between the developments of morphological structure and the digestive system functions of Percocypris pingi.【Method】Enzymatic and morphological characteristics were studied from 1 days after hatching(dah) to 46 dah in larvae and juvenile of P. pingi using enzyme linked immunosorbent assay and histological sectioning methods.【Result】Our results showed that the intestinal tract was undifferentiated in newly hatched larvae,intestinal perforation was observed at 5 dah,and the "Z" fold point of the intestinal tract appeared at 30 and 33 dah,respectively. Thereafter there was no obvious morphological changes observed. With the development of intestine,the height of the intestinal mucosal fold increased significantly,and showed a tendency of foregut>midgut>the hindgut. The results of digestive enzyme detection showed that activity of trypsin,chymotrypsin,α-amylase and lipase were firstly detected in the undifferentiated intestine,and accompanying the intestinal perforation,the activity of trypsin and chymotrypsin increased significantly,the activity of α-amylase showed no remarkable change,but the activity of lipase were decreased. Furthermore,when the first-and second-fold point were completed at 30 dah and 33 dah,the activity of the 4 digestive enzymes increased significantly. When the "Z" shaped intestine was stable,the four digestive enzymes activity showed a downward trend.【Conclusion】Under the laboratory conditions,the growth of body length and body weight was linear and trinomial relationship,respectively. The morphological development of intestine was completed at 5 dah and 35 dah,and the specific activity of digestive enzymes were highly correlated with the intestine development of P. pingi.
【Objective】The objective of this study was to reveal the relationship between the developments of morphological structure and the digestive system functions of Percocypris pingi.【Method】Enzymatic and morphological characteristics were studied from 1 days after hatching(dah) to 46 dah in larvae and juvenile of P. pingi using enzyme linked immunosorbent assay and histological sectioning methods.【Result】Our results showed that the intestinal tract was undifferentiated in newly hatched larvae,intestinal perforation was observed at 5 dah,and the "Z" fold point of the intestinal tract appeared at 30 and 33 dah,respectively. Thereafter there was no obvious morphological changes observed. With the development of intestine,the height of the intestinal mucosal fold increased significantly,and showed a tendency of foregut>midgut>the hindgut. The results of digestive enzyme detection showed that activity of trypsin,chymotrypsin,α-amylase and lipase were firstly detected in the undifferentiated intestine,and accompanying the intestinal perforation,the activity of trypsin and chymotrypsin increased significantly,the activity of α-amylase showed no remarkable change,but the activity of lipase were decreased. Furthermore,when the first-and second-fold point were completed at 30 dah and 33 dah,the activity of the 4 digestive enzymes increased significantly. When the "Z" shaped intestine was stable,the four digestive enzymes activity showed a downward trend.【Conclusion】Under the laboratory conditions,the growth of body length and body weight was linear and trinomial relationship,respectively. The morphological development of intestine was completed at 5 dah and 35 dah,and the specific activity of digestive enzymes were highly correlated with the intestine development of P. pingi.
引文
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[7]KOLKOVSKI S.Digestive enzymes in fish larvae and juveniles--implications and applications to formulated diets[J].Aquaculture,2001,200(1):181-201.
[8][8]GISBERT E,KOTZAMANIS Y.Development of digestive enzymes in common dentex Dentex dentex during early ontogeny[J].Aquaculture,2009,287(3):381-387.
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[10]刘军.长江上游特有鱼类受威胁及优先保护顺序的定量分析[J].中国环境科学,2004,24(4):395-399.
[11]邓龙君,甘维熊,曾如奎,等.雅砻江鲈鲤的人工繁殖、胚胎及卵黄囊仔鱼发育[J].水产科学,2016,35(4):393-397.
[12]易建华.不同开口饵料对胭脂鱼仔稚鱼成活率的影响[D].重庆:西南大学,2014.
[13]刘亚秋,郭忠娣,胡雨,等.大鳞副泥鳅仔稚鱼生长与消化酶活性变化[J].水生生物学报,2015,39(6):1236-1240.
[14]王永明,史晋绒,张耀光,等.人工养殖稀有鮈鲫消化道组织学观察[J].四川动物,2013,32(3):410-414.
[15][15]张臣,曾燏,彭艳,等.嘉陵江下游蛇鮈肠道形态结构及其异速生长模式[J].水产学报,2018,42(4):503-512.
[16]赵健蓉,赵月月,葛海龙,等.云南盘鮈消化系统解剖学、组织学及消化酶活性研究[J].水生生物学报,2017,41(4):853-859.
[17]张云龙,张海龙,王凌宇,等.鱼类早期发育阶段异速生长及核酸、消化酶变化的研究进展[J].中国水产科学,2017,24(3):648-656.
[18]LAZO J P,HOLT G J,ARNOLD C R.Ontogeny of pancreatic enzymes in larval red drum Sciaenops ocellatus[J].Aquaculture Nutrition,2015,6(3):183-192.
[19]LI S,WEN W,HUANG X,et al.Activities of digestive enzymes and histology of digestive system during larval development of devil stinger(Inimicus japonicus)[J].Aquaculture Research,2017,48(10):1-8.
[20]FARHOUDI A,KENARI A M A,NAZARI R M,et al.Changes of digestive enzymes activity in common carp(Cyprinus carpio)during larval ontogeny[J].Iranian Journal of Fisheries Sciences,2013,12(2):320-334.
[21]BUDDINGTON R K,DLAMOND J M.Ontogenetic development of intestinal nutrient transporters[J].Annual Review of Physiol-ogy,1989,51(4):601-617.
[22]INFANTE J L Z,CAHU C.Development and response to a diet change of some digestive enzymes in sea bass(Dicentrarchus labrax)larvae[J].Fish Physiology and Biochemistry,1994,12(5):399-408.
[23]BABAEI S S,ABEDIAN K A,NAZARI R,et al.Developmental changes of digestive enzymes in Persian sturgeon(Acipenser persicus)during larval ontogeny[J].Aquaculture,2011,318(2):138-144.
[24]SRICHANUN M,TANTIKITTI C,UTARABHAND P,et al.Gene expression and activity of digestive enzymes during the larval development of Asian seabass(Lates calcarifer)[J].Comparative Biochemistry and Physiology Part B Biochemistry&Molecular Biology,2013,165(1):1-9.
[25]MART NEZLAGOS R,TOVARRAM R D,GRACIAL P V,et al.Changes in digestive enzyme activities during larval development of leopard groupe(rMycteroperca rosacea)[J].Fish Physiology and Biochemistry,2014,40(3):1-13.
[26]PRADHAN P K,MITRA G,SOOD N,et al.Ontogeny of the digestive tract in butter catfish Ompok bimaculatus(Bloch)larvae[J].Fish Physiology and Biochemistry,2012,38(6):1601-1617.
[27]孙敏,柴学军,许源剑,等.日本黄姑鱼早期发育过程中消化酶活性变化研究[J].上海海洋大学学报,2012,21(6):965-970.
[28]GUERRERA M C,PASQUALE F D,MUGLIA U,et al.Digestive enzymatic activity during ontogenetic development in zebrafish(Danio rerio)[J].Journal of Experimental Zoology Part BMolecular and Developmental Evolution,2015,324(8):699-706.
[29]MICALE V,CARUSO G,GARAFFO M,et al.Morphological development and enzyme activities of the digestive tract in larval pandora,Pagellus erythrinus L[J].Italian Journal of Anatomy and Embryology,2006,111(1):109.
[30][30]SAVONA B,TRAMATI C,MAZZOLA A.Digestive enzymes in larvae and juveniles of farmed sharpsnout seabream(Diplodus puntazzo)(Cetti,1777)[J].Open Marine Biology Journal,2011,511(47):47-57.
[31]ALVAREZ-GONZ L C A,CERVANTES-TRUJANO M,TO-VAR-RAM R D,et al.Development of digestive enzymes in California halibut Paralichthys californicus larvae[J].Fish Physiology and Biochemistry,2005,31(1):83-93.
[32]MA H,CAHU C,ZAMBONIN J,et al.Activities of selected digestive enzymes during larval development of large yellow croaker(Pseudosciaena crocea)[J].Aquaculture,2005,245(1):239-248.
[33]张云龙,樊启学,彭聪,等.泥鳅仔稚鱼发育期间消化酶及碱性磷酸酶比活力的变化[J].淡水渔业,2013,43(1):19-23.
[34]李芹,龙勇,屈波,等.瓦氏黄颡鱼仔稚鱼发育过程中消化酶活性变化研究[J].中国水产科学,2008,15(1):73-78.
[35]MORAIS S,CAHU C,ZAMBONINO-INFANTE J L,et al.Dietary TAG source and level affect performance and lipase expression in larval sea bass(Dicentrarchus labrax)[J].Lipids,2004,39(5):449-458.
[2]ASGARI R,RAFIEE G,EAGDERI S,et al.Ontogeny of the digestive enzyme activities in hatchery produced Beluga(Huso huso)[J].Aquaculture,2013,416:33-40.
[3]LIU C X,ZHI L,TAN X Y,et al.Ontogenetic development of the digestive system in agastric chinese sucker,Myxocyprinusasiaticus,Larvae[J].Journal of the World Aquaculture Society,2013,44(3):350-362.
[4]SUZER C,KAMAC H O,COBAN D,et al.Functional changes in digestive enzyme activities of meagre(Argyrosomus regius;Asso,1801)during early ontogeny[J].Fish Physiology and Biochemistry,2013,39(4):967-977.
[5]CUI K,CHENG D,MA Z,et al.Ontogenetic development of digestive enzymes in larval and juvenile crimson snapper Lutjanus erythopterus(Bloch 1790)[J].Aquaculture Research,2017,48(8):4533-4544.
[6]GAO X Q,LIU Z F,GUAN C T,et al.Developmental changes in digestive enzyme activity in American shad,Alosa sapidissima,during early ontogeny[J].Fish Physiology&Biochemistry,2017,43(2):1-13.
[7]KOLKOVSKI S.Digestive enzymes in fish larvae and juveniles--implications and applications to formulated diets[J].Aquaculture,2001,200(1):181-201.
[8][8]GISBERT E,KOTZAMANIS Y.Development of digestive enzymes in common dentex Dentex dentex during early ontogeny[J].Aquaculture,2009,287(3):381-387.
[9]丁瑞华.四川鱼类志[M].成都:四川科学技术出版社,1994.
[10]刘军.长江上游特有鱼类受威胁及优先保护顺序的定量分析[J].中国环境科学,2004,24(4):395-399.
[11]邓龙君,甘维熊,曾如奎,等.雅砻江鲈鲤的人工繁殖、胚胎及卵黄囊仔鱼发育[J].水产科学,2016,35(4):393-397.
[12]易建华.不同开口饵料对胭脂鱼仔稚鱼成活率的影响[D].重庆:西南大学,2014.
[13]刘亚秋,郭忠娣,胡雨,等.大鳞副泥鳅仔稚鱼生长与消化酶活性变化[J].水生生物学报,2015,39(6):1236-1240.
[14]王永明,史晋绒,张耀光,等.人工养殖稀有鮈鲫消化道组织学观察[J].四川动物,2013,32(3):410-414.
[15][15]张臣,曾燏,彭艳,等.嘉陵江下游蛇鮈肠道形态结构及其异速生长模式[J].水产学报,2018,42(4):503-512.
[16]赵健蓉,赵月月,葛海龙,等.云南盘鮈消化系统解剖学、组织学及消化酶活性研究[J].水生生物学报,2017,41(4):853-859.
[17]张云龙,张海龙,王凌宇,等.鱼类早期发育阶段异速生长及核酸、消化酶变化的研究进展[J].中国水产科学,2017,24(3):648-656.
[18]LAZO J P,HOLT G J,ARNOLD C R.Ontogeny of pancreatic enzymes in larval red drum Sciaenops ocellatus[J].Aquaculture Nutrition,2015,6(3):183-192.
[19]LI S,WEN W,HUANG X,et al.Activities of digestive enzymes and histology of digestive system during larval development of devil stinger(Inimicus japonicus)[J].Aquaculture Research,2017,48(10):1-8.
[20]FARHOUDI A,KENARI A M A,NAZARI R M,et al.Changes of digestive enzymes activity in common carp(Cyprinus carpio)during larval ontogeny[J].Iranian Journal of Fisheries Sciences,2013,12(2):320-334.
[21]BUDDINGTON R K,DLAMOND J M.Ontogenetic development of intestinal nutrient transporters[J].Annual Review of Physiol-ogy,1989,51(4):601-617.
[22]INFANTE J L Z,CAHU C.Development and response to a diet change of some digestive enzymes in sea bass(Dicentrarchus labrax)larvae[J].Fish Physiology and Biochemistry,1994,12(5):399-408.
[23]BABAEI S S,ABEDIAN K A,NAZARI R,et al.Developmental changes of digestive enzymes in Persian sturgeon(Acipenser persicus)during larval ontogeny[J].Aquaculture,2011,318(2):138-144.
[24]SRICHANUN M,TANTIKITTI C,UTARABHAND P,et al.Gene expression and activity of digestive enzymes during the larval development of Asian seabass(Lates calcarifer)[J].Comparative Biochemistry and Physiology Part B Biochemistry&Molecular Biology,2013,165(1):1-9.
[25]MART NEZLAGOS R,TOVARRAM R D,GRACIAL P V,et al.Changes in digestive enzyme activities during larval development of leopard groupe(rMycteroperca rosacea)[J].Fish Physiology and Biochemistry,2014,40(3):1-13.
[26]PRADHAN P K,MITRA G,SOOD N,et al.Ontogeny of the digestive tract in butter catfish Ompok bimaculatus(Bloch)larvae[J].Fish Physiology and Biochemistry,2012,38(6):1601-1617.
[27]孙敏,柴学军,许源剑,等.日本黄姑鱼早期发育过程中消化酶活性变化研究[J].上海海洋大学学报,2012,21(6):965-970.
[28]GUERRERA M C,PASQUALE F D,MUGLIA U,et al.Digestive enzymatic activity during ontogenetic development in zebrafish(Danio rerio)[J].Journal of Experimental Zoology Part BMolecular and Developmental Evolution,2015,324(8):699-706.
[29]MICALE V,CARUSO G,GARAFFO M,et al.Morphological development and enzyme activities of the digestive tract in larval pandora,Pagellus erythrinus L[J].Italian Journal of Anatomy and Embryology,2006,111(1):109.
[30][30]SAVONA B,TRAMATI C,MAZZOLA A.Digestive enzymes in larvae and juveniles of farmed sharpsnout seabream(Diplodus puntazzo)(Cetti,1777)[J].Open Marine Biology Journal,2011,511(47):47-57.
[31]ALVAREZ-GONZ L C A,CERVANTES-TRUJANO M,TO-VAR-RAM R D,et al.Development of digestive enzymes in California halibut Paralichthys californicus larvae[J].Fish Physiology and Biochemistry,2005,31(1):83-93.
[32]MA H,CAHU C,ZAMBONIN J,et al.Activities of selected digestive enzymes during larval development of large yellow croaker(Pseudosciaena crocea)[J].Aquaculture,2005,245(1):239-248.
[33]张云龙,樊启学,彭聪,等.泥鳅仔稚鱼发育期间消化酶及碱性磷酸酶比活力的变化[J].淡水渔业,2013,43(1):19-23.
[34]李芹,龙勇,屈波,等.瓦氏黄颡鱼仔稚鱼发育过程中消化酶活性变化研究[J].中国水产科学,2008,15(1):73-78.
[35]MORAIS S,CAHU C,ZAMBONINO-INFANTE J L,et al.Dietary TAG source and level affect performance and lipase expression in larval sea bass(Dicentrarchus labrax)[J].Lipids,2004,39(5):449-458.