饲料中添加大豆皂甙对大菱鲆幼鱼生长和肠道健康的影响
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摘要
为研究大豆皂甙对大菱鲆幼鱼生长性能、消化酶活性、肠道组织结构完整性和肠道菌群结构的影响,在以鱼粉为蛋白源的基础饲料中分别添加0%和0.3%的大豆皂甙,配制成鱼粉组和大豆皂甙组2种等氮等脂的实验饲料来投喂体质量为(4.63±0.01) g的大菱鲆进行12周的摄食生长实验。结果显示,饲料中添加0.3%的大豆皂甙对大菱鲆的生长性能没有产生显著影响,但显著降低了胃蛋白酶及肠淀粉酶活性;2组实验中大菱鲆肠道组织形态无明显差异,但大豆皂甙组肠道紧密连接蛋白的基因表达量显著降低。肠道菌群分析结果显示,大菱鲆肠道中相对丰度最高的门和属分别为变形菌门和盐单胞菌属。LEfSe和MetaStat分析显示,饲料中添加大豆皂甙后显著提高了大菱鲆肠道内优势菌(变形菌门及希瓦氏菌属),皂甙水解相关的肠道微生物(鞘脂单胞菌属、普氏菌属、栖瘤胃普雷沃菌以及普通拟杆菌)及潜在致病菌(莫氏杆菌属和发光杆菌属)的相对丰度,同时显著降低了Caenimonas、Niastella和条件致病菌罗尔斯通菌属的相对丰度。研究表明,0.3%大豆皂甙抑制了大菱鲆消化酶活性及肠道紧密连接蛋白的基因表达,且引起了大菱鲆肠道菌群结构的显著改变。因此,大豆皂甙对鱼类肠道健康尤其是肠道菌群的影响不容忽视,值得进一步研究。
A 12-week feeding trial was conducted to investigate the effects of dietary soyasaponins on the growth performance, activities of digestive enzymes, intestinal morphology and intestinal bacterial community of turbot(Scophthalmus maximus). Two isonitrogenous and isolipidic experimental diets(based on 67% fish meal)containing 0% and 0.3% soyasaponins were formulated. The results showed that the growth performance was not affected by dietary soyasaponins, while the activities of intestinal pepsin and amylase were significantly depressed by dietary 0.3% soyasopnins. No obvious changes of intestinal histology were observed between two groups.However the gene expression of intestinal tight junction protein occludin was significantly declined when 0.3%soyasaponins was added. The analysis of intestinal microbiota showed that Proteobacteria and Halomonas were the most dominant phylum and genus in S. maximus intestine, respectively. LEfSe and MetaStat analysis showed that the abundance of dominant bacteria(Proteobacteria and Shewanella) was significantly increased by dietary 0.3%soyasaponins. Meanwhile, dietary soyasaponins significantly increased the abundance of intestinal saponinhydrolyzing bacteria. In addition, the abundance of some intestinal potential pathogenic bacteria such as Methylobacterium, Moraxella and Photobacterium, was increased while that of some other pathogenic bacteria such as Ralstonia was decreased by dietary soyasaponins. In conclusion, dietary 0.3% soyasaponins depressed the activities of digestive enzymes, and the gene expression of intestinal tight junction protein Occludin, and altered the intestinal microbiota of S. maximus. Thus, the effects of soyasaponins on fish intestine health, especially intestinal flora, cannot be ignored and deserve funther study.
A 12-week feeding trial was conducted to investigate the effects of dietary soyasaponins on the growth performance, activities of digestive enzymes, intestinal morphology and intestinal bacterial community of turbot(Scophthalmus maximus). Two isonitrogenous and isolipidic experimental diets(based on 67% fish meal)containing 0% and 0.3% soyasaponins were formulated. The results showed that the growth performance was not affected by dietary soyasaponins, while the activities of intestinal pepsin and amylase were significantly depressed by dietary 0.3% soyasopnins. No obvious changes of intestinal histology were observed between two groups.However the gene expression of intestinal tight junction protein occludin was significantly declined when 0.3%soyasaponins was added. The analysis of intestinal microbiota showed that Proteobacteria and Halomonas were the most dominant phylum and genus in S. maximus intestine, respectively. LEfSe and MetaStat analysis showed that the abundance of dominant bacteria(Proteobacteria and Shewanella) was significantly increased by dietary 0.3%soyasaponins. Meanwhile, dietary soyasaponins significantly increased the abundance of intestinal saponinhydrolyzing bacteria. In addition, the abundance of some intestinal potential pathogenic bacteria such as Methylobacterium, Moraxella and Photobacterium, was increased while that of some other pathogenic bacteria such as Ralstonia was decreased by dietary soyasaponins. In conclusion, dietary 0.3% soyasaponins depressed the activities of digestive enzymes, and the gene expression of intestinal tight junction protein Occludin, and altered the intestinal microbiota of S. maximus. Thus, the effects of soyasaponins on fish intestine health, especially intestinal flora, cannot be ignored and deserve funther study.
引文
[1]Krogdahl?,Bakke-McKellep A M,Baeverfjord G.Effects of graded levels of standard soybean meal on intestinal structure,mucosal enzyme activities,and pancreatic response in Atlantic salmon(Salmo salar L.)[J].Aquaculture Nutrition,2003,9(6):361-371.
[2]Francis G,Makkar H P S,Becker K.Antinutritional factors present in plant-derived alternate fish feed ingredients and their effects in fish[J].Aquaculture,2001,199(3-4):197-227.
[3]Ireland P A,Dziedzic S Z,Kearsley M W.Saponin content of soya and some commercial soya products by means of high-performance liquid chromatography of the sapogenins[J].Journal of the Science of Food and Agriculture,1986,37(7):694-698.
[4]Chen W,Ai Q H,Mai K S,et al.Effects of dietary soybean saponins on feed intake,growth performance,digestibility and intestinal structure in juvenile Japanese flounder(Paralichthys olivaceus)[J].Aquaculture,2011,318(1-2):95-100.
[5]Bureau D P,Harris A M,Cho C Y.The effects of purified alcohol extracts from soy products on feed intake and growth of chinook salmon(Oncorhynchus tshawytscha)and rainbow trout(Oncorhynchus mykiss)[J].Aquaculture,1998,161(1-4):27-43.
[6]Krogdahl A,Gajardo K,Kortner T M,et al.Soya saponins induce enteritis in Atlantic salmon(Salmo salar L.)[J].Journal of Agricultural and Food Chemistry,2015,63(15):3887-3902.
[7]Camilleri M,Madsen K,Spiller R,et al.Intestinal barrier function in health and gastrointestinal disease[J].Neurogastroenterology&Motility,2012,24(6):503-512.
[8]Kosińska A,Andlauer W.Modulation of tight junction integrity by food components[J].Food Research International,2013,54(1):951-960.
[9]Ring?E,Myklebust R,Mayhew T M,et al.Bacterial translocation and pathogenesis in the digestive tract of larvae and fry[J].Aquaculture,2007,268(1-4):251-264.
[10]Blumberg R,Powrie F.Microbiota,disease,and back to health:A metastable journey[J].Science Translational Medicine,2012,4(137):137rv7.
[11]Reveco F E,?verland M,Romarheim O H,et al.Intestinal bacterial community structure differs between healthy and inflamed intestines in Atlantic salmon(Salmo salar L.)[J].Aquaculture,2014,420-421:262-269.
[12]Li Y X,Yang P,Zhang Y J,et al.Effects of dietary glycinin on the growth performance,digestion,intestinal morphology and bacterial community of juvenile turbot,Scophthalmus maximus L.[J].Aquaculture,2017,479(1):125-133.
[13]Yang P,Hu H B,Liu Y,et al.Dietary stachyose altered the intestinal microbiota profile and improved the intestinal mucosal barrier function of juvenile turbot,Scophthalmus maximus L.[J].Aquaculture,2018,486(3):98-106.
[14]Peng M,Xu W,Ai Q H,et al.Effects of nucleotide supplementation on growth,immune responses and intestinal morphology in juvenile turbot fed diets with graded levels of soybean meal(Scophthalmus maximus L.)[J].Aquaculture,2013,392-395:51-58.
[15]AOAC.Official Methods of Analysis[M].16th ed.Washington DC:Association of Official Agricultural Chemists,1995.
[16]Anson M L.The estimation of pepsin,trypsin,papain,and cathepsin with hemoglobin[J].Journal of General Physiology,1938,22(1):79-89.
[17]Hidalgo M C,Urea E,Sanz A.Comparative study of digestive enzymes in fish with different nutritional habits.Proteolytic and amylase activities[J].Aquaculture,1999,170(3):267-283.
[18]张伟.大豆皂甙与大豆异黄酮对异育银鲫生长、生理及肠道健康的影响[D].苏州:苏州大学,2010.Zhang W.Effects of soybean saponins and soybean isoflavones on growth,physiology and intestinal health of Allogynogentic silver crucian carp[D].Suzhou:Soochow University,2010(in Chinese).
[19]S?rensen M,Penn M,El-Mowafi A,et al.Effect of stachyose,raffinose and soya-saponins supplementation on nutrient digestibility,digestive enzymes,gut morphology and growth performance in Atlantic salmon(Salmo salar,L)[J].Aquaculture,2011,314(1-4):145-152.
[20]Chikwati E M,Venold F F,Penn M H,et al.Interaction of soyasaponins with plant ingredients in diets for Atlantic salmon,Salmo salar L.[J].British Journal of Nutrition,2012,107(11):1570-1590.
[21]Liener I E.Implications of antinutritional components in soybean foods[J].Critical Reviews in Food Science and Nutrition,1994,34(1):31-67.
[22]米海峰.不同蛋白源和大豆抗营养因子对牙鲆蛋白消化酶的活性与基因表达的影响[D].青岛:中国海洋大学,2008.Mi H F.Effects of different protein sources and antinutritional factors on gene expression and activity of proteindigestive-enzyme of Japanese flounder,Paralichthys olivaceus[D].Qingdao:Ocean University of China,2008(in Chinese).
[23]陈伟.抗营养因子对牙鲆(Paralichthys olivaceus)利用大豆蛋白源的影响[D].青岛:中国海洋大学,2009.Chen W.Effects of antinutritional factors on the utilization of soybean proteins by Japanese flounder,Paralichthys olivaceus[D].Qingdao:Ocean University of China,2009(in Chinese).
[24]Gu M,Kortner T M,Penn M,et al.Effects of dietary plant meal and soya-saponin supplementation on intestinal and hepatic lipid droplet accumulation,lipoprotein and sterol metabolism in Atlantic salmon(Salmo salar L.)[J].British Journal of Nutrition,2014,111(3):432-444.
[25]Francis G,Makkar H P S,Becker K.Effects of Quillaja saponins on growth,metabolism,egg production and muscle cholesterol in individually reared Nile tilapia(Oreochromis niloticus)[J].Comparative Biochemistry and Physiology-Part C:Toxicology&Pharmacology,2001,129(2):105-114.
[26]Chavali S R,Francis T,Campbell J B.An in vitro study of immunomodulatory effects of some saponins[J].International Journal of Immunopharmacology,1987,9(6):675-683.
[27]Johnson L G.Applications of imaging techniques to studies of epithelial tight junctions[J].Advanced Drug Delivery Reviews,2005,57(1):111-121.
[28]Knudsen D,Jutfelt F,Sundh H,et al.Dietary soya saponins increase gut permeability and play a key role in the onset of soyabean-induced enteritis in Atlantic salmon(Salmo salar L.)[J].British Journal of Nutrition,2008,100(1):120-129.
[29]Kyum K M,Lee J W,Lee K Y,et al.Microbial conversion of major ginsenoside Rb1 to pharmaceutically active minor ginsenoside Rd[J].Journal of Microbiology,2005,43(5):456-462.
[30]Hu J.Characterization of soyasaponin metabolism by human gut microorganisms and bioavailability in humans[D].Ames:Iowa State University,2003.
[31]Gestetner B,Birk Y,Tencer Y.Soybean saponins.Fate of ingested soybean saponins and the physiological aspect of their hemolytic activity[J].Journal of Agricultural and Food Chemistry,1968,16(6):1031-1035.
[32]Jang I S,Kim D H.Purification and characterization ofα-L-Rhamnosidase from Bacteroides JY-6,a human intestinal bacterium[J].Biological&Pharmaceutical Bulletin,1996,19(12):1546-1549.
[33]Hasegawa H,Sung J H,Benno Y.Role of human intestinal Prevotella oris in hydrolyzing ginseng saponins[J].Planta Medica,1997,63(5):436-440.
[34]Ryan M R,Adley C C.Ralstonia spp.:emerging global opportunistic pathogens[J].European Journal of Clinical Microbiology&Infectious Diseases,2014,33(3):291-304.
[35]Khan H,Khan M A,Abdullah.Antibacterial,antioxidant and cytotoxic studies of total saponin,alkaloid and sterols contents of decoction of Joshanda:identification of components through thin layer chromatography[J].Toxicology and Industrial Health,2015,31(3):202-208.
[36]Osorio C R,Toranzo A E,Romalde J L,et al.Multiplex PCR assay for ure C and 16S rRNA genes clearly discriminates between both subspecies of Photobacterium damselae[J].Diseases of Aquatic Organisms,2000,40(3):177-183.
[37]Oldfield N J,Ala’Aldeen D A A.Neisseria and moraxella:meningitis;septicaemia;gonorrhoea;respiratory infections[M]//Greenwood D,Slack R,Barer M,et al.Medical Microbiology(Eighteenth Edition).18th ed.Edinburgh:Churchill Livingstone,2012:256-264.
[38]Maranduba C M,De Castro S B,de Souza G T,et al.Intestinal microbiota as modulators of the immune system and neuroimmune system:impact on the host health and homeostasis[J].Journal of Immunology Research,2015,2015(1):1-14.
[2]Francis G,Makkar H P S,Becker K.Antinutritional factors present in plant-derived alternate fish feed ingredients and their effects in fish[J].Aquaculture,2001,199(3-4):197-227.
[3]Ireland P A,Dziedzic S Z,Kearsley M W.Saponin content of soya and some commercial soya products by means of high-performance liquid chromatography of the sapogenins[J].Journal of the Science of Food and Agriculture,1986,37(7):694-698.
[4]Chen W,Ai Q H,Mai K S,et al.Effects of dietary soybean saponins on feed intake,growth performance,digestibility and intestinal structure in juvenile Japanese flounder(Paralichthys olivaceus)[J].Aquaculture,2011,318(1-2):95-100.
[5]Bureau D P,Harris A M,Cho C Y.The effects of purified alcohol extracts from soy products on feed intake and growth of chinook salmon(Oncorhynchus tshawytscha)and rainbow trout(Oncorhynchus mykiss)[J].Aquaculture,1998,161(1-4):27-43.
[6]Krogdahl A,Gajardo K,Kortner T M,et al.Soya saponins induce enteritis in Atlantic salmon(Salmo salar L.)[J].Journal of Agricultural and Food Chemistry,2015,63(15):3887-3902.
[7]Camilleri M,Madsen K,Spiller R,et al.Intestinal barrier function in health and gastrointestinal disease[J].Neurogastroenterology&Motility,2012,24(6):503-512.
[8]Kosińska A,Andlauer W.Modulation of tight junction integrity by food components[J].Food Research International,2013,54(1):951-960.
[9]Ring?E,Myklebust R,Mayhew T M,et al.Bacterial translocation and pathogenesis in the digestive tract of larvae and fry[J].Aquaculture,2007,268(1-4):251-264.
[10]Blumberg R,Powrie F.Microbiota,disease,and back to health:A metastable journey[J].Science Translational Medicine,2012,4(137):137rv7.
[11]Reveco F E,?verland M,Romarheim O H,et al.Intestinal bacterial community structure differs between healthy and inflamed intestines in Atlantic salmon(Salmo salar L.)[J].Aquaculture,2014,420-421:262-269.
[12]Li Y X,Yang P,Zhang Y J,et al.Effects of dietary glycinin on the growth performance,digestion,intestinal morphology and bacterial community of juvenile turbot,Scophthalmus maximus L.[J].Aquaculture,2017,479(1):125-133.
[13]Yang P,Hu H B,Liu Y,et al.Dietary stachyose altered the intestinal microbiota profile and improved the intestinal mucosal barrier function of juvenile turbot,Scophthalmus maximus L.[J].Aquaculture,2018,486(3):98-106.
[14]Peng M,Xu W,Ai Q H,et al.Effects of nucleotide supplementation on growth,immune responses and intestinal morphology in juvenile turbot fed diets with graded levels of soybean meal(Scophthalmus maximus L.)[J].Aquaculture,2013,392-395:51-58.
[15]AOAC.Official Methods of Analysis[M].16th ed.Washington DC:Association of Official Agricultural Chemists,1995.
[16]Anson M L.The estimation of pepsin,trypsin,papain,and cathepsin with hemoglobin[J].Journal of General Physiology,1938,22(1):79-89.
[17]Hidalgo M C,Urea E,Sanz A.Comparative study of digestive enzymes in fish with different nutritional habits.Proteolytic and amylase activities[J].Aquaculture,1999,170(3):267-283.
[18]张伟.大豆皂甙与大豆异黄酮对异育银鲫生长、生理及肠道健康的影响[D].苏州:苏州大学,2010.Zhang W.Effects of soybean saponins and soybean isoflavones on growth,physiology and intestinal health of Allogynogentic silver crucian carp[D].Suzhou:Soochow University,2010(in Chinese).
[19]S?rensen M,Penn M,El-Mowafi A,et al.Effect of stachyose,raffinose and soya-saponins supplementation on nutrient digestibility,digestive enzymes,gut morphology and growth performance in Atlantic salmon(Salmo salar,L)[J].Aquaculture,2011,314(1-4):145-152.
[20]Chikwati E M,Venold F F,Penn M H,et al.Interaction of soyasaponins with plant ingredients in diets for Atlantic salmon,Salmo salar L.[J].British Journal of Nutrition,2012,107(11):1570-1590.
[21]Liener I E.Implications of antinutritional components in soybean foods[J].Critical Reviews in Food Science and Nutrition,1994,34(1):31-67.
[22]米海峰.不同蛋白源和大豆抗营养因子对牙鲆蛋白消化酶的活性与基因表达的影响[D].青岛:中国海洋大学,2008.Mi H F.Effects of different protein sources and antinutritional factors on gene expression and activity of proteindigestive-enzyme of Japanese flounder,Paralichthys olivaceus[D].Qingdao:Ocean University of China,2008(in Chinese).
[23]陈伟.抗营养因子对牙鲆(Paralichthys olivaceus)利用大豆蛋白源的影响[D].青岛:中国海洋大学,2009.Chen W.Effects of antinutritional factors on the utilization of soybean proteins by Japanese flounder,Paralichthys olivaceus[D].Qingdao:Ocean University of China,2009(in Chinese).
[24]Gu M,Kortner T M,Penn M,et al.Effects of dietary plant meal and soya-saponin supplementation on intestinal and hepatic lipid droplet accumulation,lipoprotein and sterol metabolism in Atlantic salmon(Salmo salar L.)[J].British Journal of Nutrition,2014,111(3):432-444.
[25]Francis G,Makkar H P S,Becker K.Effects of Quillaja saponins on growth,metabolism,egg production and muscle cholesterol in individually reared Nile tilapia(Oreochromis niloticus)[J].Comparative Biochemistry and Physiology-Part C:Toxicology&Pharmacology,2001,129(2):105-114.
[26]Chavali S R,Francis T,Campbell J B.An in vitro study of immunomodulatory effects of some saponins[J].International Journal of Immunopharmacology,1987,9(6):675-683.
[27]Johnson L G.Applications of imaging techniques to studies of epithelial tight junctions[J].Advanced Drug Delivery Reviews,2005,57(1):111-121.
[28]Knudsen D,Jutfelt F,Sundh H,et al.Dietary soya saponins increase gut permeability and play a key role in the onset of soyabean-induced enteritis in Atlantic salmon(Salmo salar L.)[J].British Journal of Nutrition,2008,100(1):120-129.
[29]Kyum K M,Lee J W,Lee K Y,et al.Microbial conversion of major ginsenoside Rb1 to pharmaceutically active minor ginsenoside Rd[J].Journal of Microbiology,2005,43(5):456-462.
[30]Hu J.Characterization of soyasaponin metabolism by human gut microorganisms and bioavailability in humans[D].Ames:Iowa State University,2003.
[31]Gestetner B,Birk Y,Tencer Y.Soybean saponins.Fate of ingested soybean saponins and the physiological aspect of their hemolytic activity[J].Journal of Agricultural and Food Chemistry,1968,16(6):1031-1035.
[32]Jang I S,Kim D H.Purification and characterization ofα-L-Rhamnosidase from Bacteroides JY-6,a human intestinal bacterium[J].Biological&Pharmaceutical Bulletin,1996,19(12):1546-1549.
[33]Hasegawa H,Sung J H,Benno Y.Role of human intestinal Prevotella oris in hydrolyzing ginseng saponins[J].Planta Medica,1997,63(5):436-440.
[34]Ryan M R,Adley C C.Ralstonia spp.:emerging global opportunistic pathogens[J].European Journal of Clinical Microbiology&Infectious Diseases,2014,33(3):291-304.
[35]Khan H,Khan M A,Abdullah.Antibacterial,antioxidant and cytotoxic studies of total saponin,alkaloid and sterols contents of decoction of Joshanda:identification of components through thin layer chromatography[J].Toxicology and Industrial Health,2015,31(3):202-208.
[36]Osorio C R,Toranzo A E,Romalde J L,et al.Multiplex PCR assay for ure C and 16S rRNA genes clearly discriminates between both subspecies of Photobacterium damselae[J].Diseases of Aquatic Organisms,2000,40(3):177-183.
[37]Oldfield N J,Ala’Aldeen D A A.Neisseria and moraxella:meningitis;septicaemia;gonorrhoea;respiratory infections[M]//Greenwood D,Slack R,Barer M,et al.Medical Microbiology(Eighteenth Edition).18th ed.Edinburgh:Churchill Livingstone,2012:256-264.
[38]Maranduba C M,De Castro S B,de Souza G T,et al.Intestinal microbiota as modulators of the immune system and neuroimmune system:impact on the host health and homeostasis[J].Journal of Immunology Research,2015,2015(1):1-14.