β-葡聚糖对杀鲑气单胞菌感染后虹鳟应激过程的调节作用
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摘要
为研究不同浓度的β-葡聚糖对感染杀鲑气单胞菌(Aeromonas salmonicida)的虹鳟(Oncorhynchus mykiss)应激过程的调节作用,在虹鳟饲料中添加不同浓度的β-葡聚糖(0,0.05%,0.1%,和0.2%),投喂42 d后感染杀鲑气单胞菌,在感染后第2,4和6天取样检测血清应激指标的变化,第7天时统计虹鳟的存活率。结果表明,添加β-葡聚糖能显著提高感染杀鲑气单胞菌后虹鳟的存活率,其中0.2%剂量的保护效果最好(P<0.05)。感染后第4天β-葡聚糖组的血清总蛋白(total protein,TP)浓度显著高于对照组(P<0.05)。0.1%和0.2%组的血清丙二醛(malondialdehyde,MDA)活性在感染后第2天达到峰值,早于0.05%和对照组。0.1%和0.2%组血清髓过氧化物酶(myeloperoxidase,MPO)活性在感染后显著上升,而0.05%和对照组却显著下降(P<0.05)。β-葡聚糖组在感染后血清乳酸脱氢酶(lactic dehydrogenase,LDH)活性一直显著低于对照组(P<0.05)。β-葡聚糖组血清甘油三酯(triglycerides,TG)和总胆固醇(total cholesterol,TCH)感染后下降程度显著低于对照组(P<0.05)。感染后第4天,β-葡聚糖组的血清尿素氮(urea nitrogen,BUN)浓度显著低于对照组(P<0.05)。总胆红素(total bilirubin,T-BIL)浓度在感染后显著下降(P<0.05),其中0.2%组下降幅度最小。各组肝脏和脾脏热休克蛋白70基因(heat shock protein 70,HSP70)表达量在感染后都显著升高(P<0.05),0.1%和0.2%组表达峰值出现时间早于其他组,且其峰值高于其他组峰值。综上所述,本实验中,添加β-葡聚糖能显著提高感染杀鲑气单胞菌的虹鳟的存活率和有效减弱细菌感染引起的机体应激反应,其中0.2%剂量的保护效果最好且对应激过程的减弱效果最明显(P<0.05)。
In recent years, immunostimulants have become a new research field in aquaculture. β-glucan is one of the most important immunostimulants, which can enhance the body's resistance to infection by improving the non-specific and cellular immunity of animals. The present study evaluated the effects of different dietary β-glucan dosages(0, 0.05%, 0.1%, and 0.2%) on the anti-infection ability of rainbow trout(Oncorhynchus mykiss) after 42 days of feeding. Thereafter, rainbow trout were infected with Aeromonas salmonicida and sampled on the 2 nd, 4 th, and 6 th days post-infection. Survival rates as well as the regulatory processes of serum stress-related factors were analyzed. Survival rates in the β-glucan groups increased significantly compared with those in the control group after A. salmonicida infection, with the 0.2% group showing the best effect(P<0.05). On the 4 th day after infection, the serum total protein concentrations in all β-glucan groups were significantly higher than those in the control group(P<0.05). Serum malondialdehyde activity in the 0.1% and 0.2% groups reached a peak on the 2 nd day post-infection, which was earlier than in the other infected groups. The activity of serum myeloperoxidase in the 0.1% and 0.2% groups increased significantly after infection, whereas in the 0.05% and control groups, it decreased significantly(P<0.05). The activity of lactate dehydrogenase in all β-glucan groups after infection was significantly lower than that in the control group(P<0.05). Serum triglyceride and total cholesterol levels in all β-glucan groups increased more obviously than those in the control group(P<0.05). On the 4 th day after infection, the serum urea nitrogen concentration in all β-glucan groups was markedly lower than that in the infected control group(P<0.05). Total bilirubin(T-BIL) concentration decreased significantly after infection; however, T-BIL in the 0.2% group was clearly higher than that in the other groups. After infection, HSP70 expression in the liver and spleen in all groups increased significantly. In the 0.1% and 0.2% groups, it reached a higher peak earlier than in the control group. Overall, the liver and head kidney in rainbow trout were damaged to a certain extent after infection with A. salmonicida; however, β-glucan can effectively protect fish from damage caused by A. salmonicida, with 0.2% β-glucan showing the best effect.
In recent years, immunostimulants have become a new research field in aquaculture. β-glucan is one of the most important immunostimulants, which can enhance the body's resistance to infection by improving the non-specific and cellular immunity of animals. The present study evaluated the effects of different dietary β-glucan dosages(0, 0.05%, 0.1%, and 0.2%) on the anti-infection ability of rainbow trout(Oncorhynchus mykiss) after 42 days of feeding. Thereafter, rainbow trout were infected with Aeromonas salmonicida and sampled on the 2 nd, 4 th, and 6 th days post-infection. Survival rates as well as the regulatory processes of serum stress-related factors were analyzed. Survival rates in the β-glucan groups increased significantly compared with those in the control group after A. salmonicida infection, with the 0.2% group showing the best effect(P<0.05). On the 4 th day after infection, the serum total protein concentrations in all β-glucan groups were significantly higher than those in the control group(P<0.05). Serum malondialdehyde activity in the 0.1% and 0.2% groups reached a peak on the 2 nd day post-infection, which was earlier than in the other infected groups. The activity of serum myeloperoxidase in the 0.1% and 0.2% groups increased significantly after infection, whereas in the 0.05% and control groups, it decreased significantly(P<0.05). The activity of lactate dehydrogenase in all β-glucan groups after infection was significantly lower than that in the control group(P<0.05). Serum triglyceride and total cholesterol levels in all β-glucan groups increased more obviously than those in the control group(P<0.05). On the 4 th day after infection, the serum urea nitrogen concentration in all β-glucan groups was markedly lower than that in the infected control group(P<0.05). Total bilirubin(T-BIL) concentration decreased significantly after infection; however, T-BIL in the 0.2% group was clearly higher than that in the other groups. After infection, HSP70 expression in the liver and spleen in all groups increased significantly. In the 0.1% and 0.2% groups, it reached a higher peak earlier than in the control group. Overall, the liver and head kidney in rainbow trout were damaged to a certain extent after infection with A. salmonicida; however, β-glucan can effectively protect fish from damage caused by A. salmonicida, with 0.2% β-glucan showing the best effect.
引文
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[8]Sirimanapong W,Adams A,Ooi E L,et al.The effects of feeding immunostimulantβ-glucan on the immune response of Pangasianodon hypophthalmus[J].Fish and Shellfish Immunology,2015,45(2):357-366.
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[14]Xu X J,Xu B,Wang J,et al.Studies on blood chemistry indices and histopathology of Pseudosciaena crocea artificially challenged with Vibrio harveyi[J].Journal of fisheries of China,2010,34(4):618-625.[徐晓津,徐斌,王军,等.大黄鱼感染哈维氏弧菌后血液生化指标的变化及组织病理学观察[J].水产学报,2010,34(4):618-625.]
[15]Farombi E O,Adelowo O A,Ajimoko Y R.Biomarkers of oxidative stress and heavy metal levels as indicators of environmental pollution in African cat fish(Clarias gariepinus)from Nigeria Ogun River[J].International Journal of Environmental Research and Public health,2007,4(2):158-165.
[16]Heise K,Puntarulo S,Nikinmaa M,et al.Oxidative stress and HIF-1 DNA binding during stressful cold exposure and recovery in the North Sea eelpout(Zoarces viviparus)[J].Comparative Biochemistry and Physiology Part A:Molecular and Integrative Physiology,2006,143(4):494-503.
[17]Han L L,Shen X L.Study Progress of myeloperoxidase and its gene polymorphism[J].Medical Recapitulate,2007,13(17):1288-1290.[韩莉莉,沈晓丽.髓过氧化物酶及其基因多态性的研究进展[J].医学综述,2007,13(17):1288-1290.]
[18]Liu T F,Ma R R,Xiao Y Y,et al.Correlation of the relative expression of myeloperoxidase and praziquantel concentration in blood of the crucian carp,Carassius auratus[J].Journal of Fishery Sciences of China,2016,23(6):1332-1338.[刘腾飞,马荣荣,肖艳翼,等.鲫血髓过氧化物酶的表达及其与血药浓度的关联性[J].中国水产科学,2016,23(6):1332-1338.]
[19]Chang Y M,Cao D C,Sun X W,et al.Changes of serum biochemical indices of common carp affected by cold temperatures[J].Chinese Journal of Fisheries,2006,19(2):71-75.[常玉梅,曹鼎臣,孙效文,等.低温胁迫对鲤血清生化指标的影响[J].水产学杂志,2006,19(2):71-75.]
[20]Ohtsuka K,Hata M.Molecular chaperone function of mammalian Hsp70 and Hsp40–a review[J].International Journal of Hyperthermia,2000,16(3):231-245.
[21]Cheng P,Liu X,Zhang G,et al.Cloning and expression analysis of a HSP70 gene from Pacific abalone(Haliotis discus hannai)[J].Fish and Shellfish Immunology,2007,22(1-2):77-87.
[22]Deane E E,Li J,Woo N Y.Modulated heat shock protein expression during pathogenic Vibrio alginolyticus stress of sea bream[J].Diseases of Aquatic Organisms,2004,62(3):205-215.
[23]Liu B,Xu L,Ge X,et al.Effects of mannan oligosaccharide on the physiological responses,HSP70 gene expression and disease resistance of Allogynogenetic crucian carp(Carassius auratus gibelio)under Aeromonas hydrophila infection[J].Fish and Shellfish Immunology,2013,34(6):1395-1403.
[2]Rawles S D,Kocabas A,Gatlin D M,et al.Dietary supplementation of terramycin and Romet-30 does not enhance growth of channel catfish but does influence tissue residues[J].Journal of the World Aquaculture Society,1997,28(4):392-401.
[3]Alderman D J,Hastings T S.Antibiotic use in aquaculture:development of antibiotic resistance–potential for consumer health risks[J].International Journal of Food Science and Technology,1998,33(2):139-155.
[4]Patterson J A,Burkholder K M.Application of prebiotics and probiotics in poultry production[J].Poultry Science,2003,82(4):627-631.
[5]Sakai M.Current research status of fish immunostimulants[J].Aquaculture,1999,172(1):63-92.
[6]Misra C K,Das B K,Mukherjee S C,et al.Effect of long term administration of dietaryβ-glucan on immunity,growth and survival of Labeo rohita fingerlings[J].Aquaculture,2006,255(1):82-94.
[7]Lin S,Pan Y,Luo L,et al.Effects of dietaryβ-1,3-glucan,chitosan or raffinose on the growth,innate immunity and resistance of koi(Cyprinus carpio koi)[J].Fish and Shellfish Immunology,2011,31(6):788-794.
[8]Sirimanapong W,Adams A,Ooi E L,et al.The effects of feeding immunostimulantβ-glucan on the immune response of Pangasianodon hypophthalmus[J].Fish and Shellfish Immunology,2015,45(2):357-366.
[9]Siwicki A K,Anderson D P,Rumsey G L.Dietary intake of immunostimulants by rainbow trout affects non-specific immunity and protection against furunculosis[J].Veterinary Immunology and immunopathology,1994,41(1-2):125-139.
[10]Irianto A,Austin B.Use of probiotics to control furunculosis in rainbow trout,Oncorhynchus mykiss(Walbaum)[J].Journal of Fish Diseases,2002,25(6):333-342.
[11]Gopalakannan A,Arul V.Enhancement of the innate immunopathdg,immune system and disease-resistant activity in Cyprinus carpio by oral administration ofβ-glucan and whole cell yeast[J].Aquaculture Research,2010,41(6):884-892.
[12]Lauridsen J H,Buchmann K.Effects of short-and long-term glucan feeding of rainbow trout(Salmonidae)on the susceptibility to Ichthyophthirius multifiliis infections[J].Acta Ichthyologica et Piscatoria,2010,40(1):61-66.
[13]Li Y J,Wang W M,Huang Y H,et al.Effects of dietaryβ-glucan on growth performance,serum biochemical indices and resistance against Aeromonas hydrophila of juvenile yellow catfish(Pseudobagrus fulvidraco)[J].Animal Nutrition,2015,27(12):3754-3762.[李永娟,王卫民,黄燕华,等.饲料中添加β-葡聚糖对黄颡鱼幼鱼生长性能,血清生化指标及抗嗜水气单胞菌感染能力的影响[J].动物营养学报,2015,27(12):3754-3762.]
[14]Xu X J,Xu B,Wang J,et al.Studies on blood chemistry indices and histopathology of Pseudosciaena crocea artificially challenged with Vibrio harveyi[J].Journal of fisheries of China,2010,34(4):618-625.[徐晓津,徐斌,王军,等.大黄鱼感染哈维氏弧菌后血液生化指标的变化及组织病理学观察[J].水产学报,2010,34(4):618-625.]
[15]Farombi E O,Adelowo O A,Ajimoko Y R.Biomarkers of oxidative stress and heavy metal levels as indicators of environmental pollution in African cat fish(Clarias gariepinus)from Nigeria Ogun River[J].International Journal of Environmental Research and Public health,2007,4(2):158-165.
[16]Heise K,Puntarulo S,Nikinmaa M,et al.Oxidative stress and HIF-1 DNA binding during stressful cold exposure and recovery in the North Sea eelpout(Zoarces viviparus)[J].Comparative Biochemistry and Physiology Part A:Molecular and Integrative Physiology,2006,143(4):494-503.
[17]Han L L,Shen X L.Study Progress of myeloperoxidase and its gene polymorphism[J].Medical Recapitulate,2007,13(17):1288-1290.[韩莉莉,沈晓丽.髓过氧化物酶及其基因多态性的研究进展[J].医学综述,2007,13(17):1288-1290.]
[18]Liu T F,Ma R R,Xiao Y Y,et al.Correlation of the relative expression of myeloperoxidase and praziquantel concentration in blood of the crucian carp,Carassius auratus[J].Journal of Fishery Sciences of China,2016,23(6):1332-1338.[刘腾飞,马荣荣,肖艳翼,等.鲫血髓过氧化物酶的表达及其与血药浓度的关联性[J].中国水产科学,2016,23(6):1332-1338.]
[19]Chang Y M,Cao D C,Sun X W,et al.Changes of serum biochemical indices of common carp affected by cold temperatures[J].Chinese Journal of Fisheries,2006,19(2):71-75.[常玉梅,曹鼎臣,孙效文,等.低温胁迫对鲤血清生化指标的影响[J].水产学杂志,2006,19(2):71-75.]
[20]Ohtsuka K,Hata M.Molecular chaperone function of mammalian Hsp70 and Hsp40–a review[J].International Journal of Hyperthermia,2000,16(3):231-245.
[21]Cheng P,Liu X,Zhang G,et al.Cloning and expression analysis of a HSP70 gene from Pacific abalone(Haliotis discus hannai)[J].Fish and Shellfish Immunology,2007,22(1-2):77-87.
[22]Deane E E,Li J,Woo N Y.Modulated heat shock protein expression during pathogenic Vibrio alginolyticus stress of sea bream[J].Diseases of Aquatic Organisms,2004,62(3):205-215.
[23]Liu B,Xu L,Ge X,et al.Effects of mannan oligosaccharide on the physiological responses,HSP70 gene expression and disease resistance of Allogynogenetic crucian carp(Carassius auratus gibelio)under Aeromonas hydrophila infection[J].Fish and Shellfish Immunology,2013,34(6):1395-1403.