Cu~(2+)富集与释放对蚂蟥生长、内在品质及相关基因表达影响的研究
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摘要
该研究旨在探究Cu~(2+)富集与释放对蚂蟥生长和内在品质影响,以及蚂蟥不同组织对Cu~(2+)富集与释放规律。在安全浓度范围内,在蚂蟥日常养殖中添加适宜浓度Cu~(2+),养殖50 d后停止添加Cu~(2+)继续养殖50 d,测定不同时期蚂蟥生长指标、不同组织Cu~(2+)含量、以及内在品质变化。结果表明,0.014 mg·L-1Cu~(2+)能显著促进蚂蟥生长(P<0.05),降低死亡率;在Cu~(2+)富集初期40 d内蚂蟥特定生长率、消化酶活性、生长与消化酶相关基因表达量显著高于对照组(P<0.05),释放期无明显差异;Cu~(2+)组蚂蟥免疫酶活性、免疫酶基因表达量与对照组比呈现先升高后降低的趋势(P<0.05);蚂蟥不同组织对Cu~(2+)富集能力为消化道>肌肉>皮肤,释放能力为肌肉>消化道>皮肤;Cu~(2+)组蚂蟥内在品质、水蛭素基因表达量与对照组无显著差异。综上,Cu~(2+)能提高蚂蟥消化酶、抗逆酶活性及相关基因的表达,促进蚂蟥生长、提高免疫能力,但并不影响药材水蛭内在品质,蚂蟥至少需要30 d才能完全释放体内富集的Cu~(2+)。
The present study explored the effects of Cu~(2+) enrichment and release on the growth and internal quality of Whitmania pigra,and the regularity of enrichment and release in different tissues of Wh. pigra. In the range of safe concentration( SC),a certain concentration of Cu~(2+) was added to the Wh. pigra for 50 days,and stopped adding for another 50 days. The growth index of Wh.pigra,Cu~(2+) content in different tissues,and the internal quality were determined. The results showed that the average body weight of Wh. pigra in 0. 014 mg·L-1 of Cu~(2+) group was significantly higher than that in control group during the experimental period( P <0. 05),and the mortality rate was lower than that in the control group. The specific growth rate,digestive enzyme activity,growth and digestive enzyme related gene expression of Wh. pigra in Cu~(2+) group were significantly higher than that in the control group( P <0. 05) during the early 40 days of enrichment,and there was no significant difference in the release period. In Cu~(2+) group,the expression of immune enzyme and immune enzyme related gene of Wh. pigra increased first and then decreased compared with control group( P < 0. 05). The ability of enriched Cu~(2+) in different tissues of Wh. pigra was digestive tract > muscle > skin. The ability of release Cu~(2+) was muscle > digestive tract > skin. There was no significant difference in the internal quality and hirudin gene expression between Cu~(2+) group and control group. In conclusion,Cu~(2+) can improve the expression of digestive enzymes,immune enzyme and related genes of Wh. pigra,promote the growth of Wh. pigra and enhance their immunity,but it doesn't affect the internal quality of Wh.pigra. The Wh. pigra can completely release Cu~(2+) within 30 days.
The present study explored the effects of Cu~(2+) enrichment and release on the growth and internal quality of Whitmania pigra,and the regularity of enrichment and release in different tissues of Wh. pigra. In the range of safe concentration( SC),a certain concentration of Cu~(2+) was added to the Wh. pigra for 50 days,and stopped adding for another 50 days. The growth index of Wh.pigra,Cu~(2+) content in different tissues,and the internal quality were determined. The results showed that the average body weight of Wh. pigra in 0. 014 mg·L-1 of Cu~(2+) group was significantly higher than that in control group during the experimental period( P <0. 05),and the mortality rate was lower than that in the control group. The specific growth rate,digestive enzyme activity,growth and digestive enzyme related gene expression of Wh. pigra in Cu~(2+) group were significantly higher than that in the control group( P <0. 05) during the early 40 days of enrichment,and there was no significant difference in the release period. In Cu~(2+) group,the expression of immune enzyme and immune enzyme related gene of Wh. pigra increased first and then decreased compared with control group( P < 0. 05). The ability of enriched Cu~(2+) in different tissues of Wh. pigra was digestive tract > muscle > skin. The ability of release Cu~(2+) was muscle > digestive tract > skin. There was no significant difference in the internal quality and hirudin gene expression between Cu~(2+) group and control group. In conclusion,Cu~(2+) can improve the expression of digestive enzymes,immune enzyme and related genes of Wh. pigra,promote the growth of Wh. pigra and enhance their immunity,but it doesn't affect the internal quality of Wh.pigra. The Wh. pigra can completely release Cu~(2+) within 30 days.
引文
[1]中国药典.一部[S].2015:77.
[2]史红专,刘飞,郭巧生.宽体金线蛭耗氧率与窒息点的初步研究[J].中国中药杂志,2005,30(23):1817.
[3]史红专,刘宏,郭巧生,等.不同月龄蚂蟥消化酶活性的研究[J].中国中药杂志,2015,40(14):2796.
[4]史红专,刘飞,郭巧生.温度对蚂蟥生长及摄食规律影响的初步研究[J].中国中药杂志,2006,31(23):1944.
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[7]Liu F,Guo Q,Shi H,et al.Genetic diversity analysis of Perinereis aibuhitensis,based on ISSR and SRAP markers of Chinese coast populations[J].Biochem Syst Ecol,2014,57:262.
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[9]李蒙蒙,史红专,郭巧生,等.蚂蟥与螺蛳耗氧率、窒息点相关性及最适投喂方式的研究[J].中国中药杂志,2016,41(15):2790.
[10]冼健安,王安利.饲料中铜离子对斑节对虾血细胞组成、活性和免疫功能的影响[J].海洋科学,2013,37(12):40.
[11]关海红,潘伟志,蔺玉华,等.4种重金属对松浦鲤生长及组织的影响[J].华中农业大学学报,2005,24(1):68.
[12]段晓姣.生态沟渠中梨形环棱螺的食性及食物选择的研究[D].武汉:华中农业大学,2013.
[13]袁维佳,俞膺浩.螺蛳对重金属元素的富集作用[J].上海师范大学学报:自然科学版,2000,29(3):73.
[14]孟凡国,史红专,王嘉,等.蚂蟥与梨形环棱螺营养成分的相关性研究[J].中国中药杂志,2013,38(12):1915.
[15]姜云霞.微量元素铜的研究进展及其对动物健康的影响[J].微量元素与健康研究,2007(5):58.
[16]张传永,刘庆,陈燕妮.重金属对水生生物毒性作用研究进展[J].生命科学仪器,2008(11):3.
[17]Rainbow P S.海洋生物对重金属的积累及意义[J].海洋环境科学,1992(1):44.
[18]史红专,施国伟,郭巧生,等.铜锌铅重金属离子对蚂蟥急性毒性的研究[J].中药材,2018(6):1276.
[19]胡琼英,秦春,陈敏.生物化学与分子生物学实验技术[M].3版.北京:化学工业出版社,2014:28.
[20]郑碧玉.改良的连苯三酚自氧化测定超氧化物歧化酶活性的方法[J].生物化学与生物物理进展,1991,18(2):163.
[21]桂远明.水产养殖学专业实验实习教材水产动物机能学实验[M].北京:中国农业出版社,2004:124.
[22]芶琳,单志.生物化学实验[M].成都:西南交通大学出版社,2010:103.
[23]刘存岐,王安利,王维娜,等.海水中几种金属离子对中国对虾幼体体内碱性磷酸酶和ATPase的影响[J].水产学报,2001,25(4):298.
[24]王维娜,王安利,孙儒泳.水环境中的铜锌铁钴离子对日本沼虾消化酶和碱性磷酸酶的影响[J].动物学报,2001(S1):72.
[25]章龙珍,姚志峰,庄平,等.铜暴露对中华鲟幼鱼的生长、消化酶活性和组织中铜积累的影响[J].复旦学报:自然科学版,2012,51(6):789.
[26]Mikel W B,Althen T G,Rogers R W,et al.Effects of exogenous porcine somatotropin on the carcass composition,hormonal and metabolic profiles,lipogenic capacity,and binding of insulin to erythrocyte receptors of fast-versus slow-growing swine[J].J Anim Sci,1993,71(7):1786.
[27]陶志云,徐文娟,宋迟,等.鸭胚胎期和发育早期垂体GH基因表达的发育性变化分析[J].畜牧与兽医,2012(S1):199.
[28]闫云峰,杨华,杨永林,等.日粮不同蛋白质水平对绵羊IGF-1和GH分泌及基因表达的影响[J].畜牧兽医学报,2015(1):99.
[29]Mata-Sotres J A,Moyano F J,Martínez-Rodríguez G,et al.Daily rhythms of digestive enzyme activity and gene expression in gilthead seabream(Sparus aurata)during ontogeny[J].Comp Biochem Physiol A Mol Integr Physiol,2016,197:43.
[30]Lazarova N,Krumova E,Stefanova T,et al.The oxidative stress response of the filamentous yeast Trichosporon cutaneum R57 to copper,cadmium and chromium exposure[J].Biotechnol Biotechnol Equip,2014,28(5):855.
[31]周健恺,徐善良,邱成功,等.Cu2+急性毒性下银鲳幼鱼红细胞核异常和抗氧化酶活性的变化[J].水产学报,2014,38(8):1106.
[32]宋波澜,张华.Cu离子浓度对杂交鲟幼鱼行为、血液指标和抗氧化酶的影响[J].大连海洋大学学报,2014,29(3):241.
[33]Peng C,Zhao X G,Han Y,et al.Toxic effects of chronic sublethal Cu2+,Pb2+and Cd2+on antioxidant enzyme activities in various tissues of the blood cockle,Anadara granosa[J].J Residuals Sci Tech,2015,12(3):125.
[34]Parihar M S,Tarangini J,Taruna H,et al.Responseof superoxide dismutase,glutathione peroxidase and reduced glutathione antioxidant defenses in gills of the freshwater catfish(Heteropneustes fossilis)to short-term elevated temperature[J].J Therm Biol,1997,22(3):151.
[35]励建荣,李学鹏,王丽,等.贝类对重金属的吸收转运与累积规律研究进展[J].水产科学,2007,26(1):51.
[36]杨志彪,赵云龙,周忠良,等.水体铜对中华绒螯蟹体内铜分布和消化酶活性的影响[J].水产学报,2005,29(4):497.
[2]史红专,刘飞,郭巧生.宽体金线蛭耗氧率与窒息点的初步研究[J].中国中药杂志,2005,30(23):1817.
[3]史红专,刘宏,郭巧生,等.不同月龄蚂蟥消化酶活性的研究[J].中国中药杂志,2015,40(14):2796.
[4]史红专,刘飞,郭巧生.温度对蚂蟥生长及摄食规律影响的初步研究[J].中国中药杂志,2006,31(23):1944.
[5]史红专,郭巧生,朱再标,等.不同温度和p H对蚂蟥消化道酶活性影响的初步研究[J].中国中药杂志,2012,37(17):2538.
[6]Liu F,Guo Q,Shi H,et al.Genetic diversity and phylogenetic relationships among and within populations of Whitmania pigra,and Hirudo nipponica,based on ISSR and SRAP markers[J].Biochem Syst Ecol,2013,51(51):215.
[7]Liu F,Guo Q,Shi H,et al.Genetic diversity analysis of Perinereis aibuhitensis,based on ISSR and SRAP markers of Chinese coast populations[J].Biochem Syst Ecol,2014,57:262.
[8]李蒙蒙,郭巧生,史红专,等.不同养殖密度和换水频率对蚂蟥生长和内在品质影响的研究[J].中国中药杂志,2016,41(6):995.
[9]李蒙蒙,史红专,郭巧生,等.蚂蟥与螺蛳耗氧率、窒息点相关性及最适投喂方式的研究[J].中国中药杂志,2016,41(15):2790.
[10]冼健安,王安利.饲料中铜离子对斑节对虾血细胞组成、活性和免疫功能的影响[J].海洋科学,2013,37(12):40.
[11]关海红,潘伟志,蔺玉华,等.4种重金属对松浦鲤生长及组织的影响[J].华中农业大学学报,2005,24(1):68.
[12]段晓姣.生态沟渠中梨形环棱螺的食性及食物选择的研究[D].武汉:华中农业大学,2013.
[13]袁维佳,俞膺浩.螺蛳对重金属元素的富集作用[J].上海师范大学学报:自然科学版,2000,29(3):73.
[14]孟凡国,史红专,王嘉,等.蚂蟥与梨形环棱螺营养成分的相关性研究[J].中国中药杂志,2013,38(12):1915.
[15]姜云霞.微量元素铜的研究进展及其对动物健康的影响[J].微量元素与健康研究,2007(5):58.
[16]张传永,刘庆,陈燕妮.重金属对水生生物毒性作用研究进展[J].生命科学仪器,2008(11):3.
[17]Rainbow P S.海洋生物对重金属的积累及意义[J].海洋环境科学,1992(1):44.
[18]史红专,施国伟,郭巧生,等.铜锌铅重金属离子对蚂蟥急性毒性的研究[J].中药材,2018(6):1276.
[19]胡琼英,秦春,陈敏.生物化学与分子生物学实验技术[M].3版.北京:化学工业出版社,2014:28.
[20]郑碧玉.改良的连苯三酚自氧化测定超氧化物歧化酶活性的方法[J].生物化学与生物物理进展,1991,18(2):163.
[21]桂远明.水产养殖学专业实验实习教材水产动物机能学实验[M].北京:中国农业出版社,2004:124.
[22]芶琳,单志.生物化学实验[M].成都:西南交通大学出版社,2010:103.
[23]刘存岐,王安利,王维娜,等.海水中几种金属离子对中国对虾幼体体内碱性磷酸酶和ATPase的影响[J].水产学报,2001,25(4):298.
[24]王维娜,王安利,孙儒泳.水环境中的铜锌铁钴离子对日本沼虾消化酶和碱性磷酸酶的影响[J].动物学报,2001(S1):72.
[25]章龙珍,姚志峰,庄平,等.铜暴露对中华鲟幼鱼的生长、消化酶活性和组织中铜积累的影响[J].复旦学报:自然科学版,2012,51(6):789.
[26]Mikel W B,Althen T G,Rogers R W,et al.Effects of exogenous porcine somatotropin on the carcass composition,hormonal and metabolic profiles,lipogenic capacity,and binding of insulin to erythrocyte receptors of fast-versus slow-growing swine[J].J Anim Sci,1993,71(7):1786.
[27]陶志云,徐文娟,宋迟,等.鸭胚胎期和发育早期垂体GH基因表达的发育性变化分析[J].畜牧与兽医,2012(S1):199.
[28]闫云峰,杨华,杨永林,等.日粮不同蛋白质水平对绵羊IGF-1和GH分泌及基因表达的影响[J].畜牧兽医学报,2015(1):99.
[29]Mata-Sotres J A,Moyano F J,Martínez-Rodríguez G,et al.Daily rhythms of digestive enzyme activity and gene expression in gilthead seabream(Sparus aurata)during ontogeny[J].Comp Biochem Physiol A Mol Integr Physiol,2016,197:43.
[30]Lazarova N,Krumova E,Stefanova T,et al.The oxidative stress response of the filamentous yeast Trichosporon cutaneum R57 to copper,cadmium and chromium exposure[J].Biotechnol Biotechnol Equip,2014,28(5):855.
[31]周健恺,徐善良,邱成功,等.Cu2+急性毒性下银鲳幼鱼红细胞核异常和抗氧化酶活性的变化[J].水产学报,2014,38(8):1106.
[32]宋波澜,张华.Cu离子浓度对杂交鲟幼鱼行为、血液指标和抗氧化酶的影响[J].大连海洋大学学报,2014,29(3):241.
[33]Peng C,Zhao X G,Han Y,et al.Toxic effects of chronic sublethal Cu2+,Pb2+and Cd2+on antioxidant enzyme activities in various tissues of the blood cockle,Anadara granosa[J].J Residuals Sci Tech,2015,12(3):125.
[34]Parihar M S,Tarangini J,Taruna H,et al.Responseof superoxide dismutase,glutathione peroxidase and reduced glutathione antioxidant defenses in gills of the freshwater catfish(Heteropneustes fossilis)to short-term elevated temperature[J].J Therm Biol,1997,22(3):151.
[35]励建荣,李学鹏,王丽,等.贝类对重金属的吸收转运与累积规律研究进展[J].水产科学,2007,26(1):51.
[36]杨志彪,赵云龙,周忠良,等.水体铜对中华绒螯蟹体内铜分布和消化酶活性的影响[J].水产学报,2005,29(4):497.