四种正交子一代奥尼罗非鱼耐链球菌病能力差异研究
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摘要
链球菌病是我国罗非鱼(Oreochromis spp.)产业的重大病害,迄今为止尚无有效防治办法,提高鱼体内在抗病力,进行品种改良是解决该病害的重要途径。本研究以尼罗罗非鱼(O. niloticus)埃及品系(Egypt strain, NE)♀、尼罗罗非鱼湘湖品系(Xianghu strain, NX)♀、奥利亚罗非鱼(O. aureus)"夏奥1号"(Xia'ao strain, AX)♂和奥利亚罗非鱼埃及品系(Egypt strain, AE)♂为亲本进行正交配组,获得NE♀×AX♂(EX)、NX♀×AX♂(XX)、NX♀×AE♂(XE)和NE♀×AE♂(EE)4种杂交组合。4种杂交子一代奥尼罗非鱼加强培育100 d后,进行无乳链球菌(Streptococcus agalactiae)人工感染实验,通过感染后的相对存活率和血液生化指标变化的比较来评价其耐链球菌病能力。结果显示:感染后7 d的相对存活率EX奥尼罗非鱼(50%)>EE奥尼罗非鱼(37.5%)>XE奥尼罗非鱼(33.3%)>XX奥尼罗非鱼(20.8%);4种奥尼鱼中只有EX奥尼罗非鱼的丙氨酸氨基转移酶(alanine aminotransferase, ALT)感染前后没有显著性变化(P>0.05);4种奥尼罗非鱼K+、Na+、C1-和Ca2+等电解质离子的含量感染后都发生显著性变化(P<0.05),但EX奥尼罗非鱼的Na+含量、Ca2+含量和Cl-含量在感染后3 d恢复到感染前水平;4种奥尼罗非鱼的球蛋白(globulin,GLO)在感染后3 d显著上升(P<0.05),并在感染后7 d达到峰值,此时ALB/GLO都显著降低(P<0.05);4种奥尼罗非鱼的乳酸脱氢酶(lactate dehydrogenase, LDH)在感染后显著增高(P<0.05),感染后的峰值分别为感染前的1.81倍、6.1倍、2.42倍和3.77倍;感染前EE奥尼罗非鱼的超氧化物歧化酶(superoxide dismutase, SOD)显著低于其余3种奥尼罗非鱼的SOD,而感染后1 d和7 d EX奥尼罗非鱼的SOD最低;4种奥尼罗非鱼的甘油三酯(triglyceride, TG)都在感染后3 d显著下降(P<0.05),EX、XX和XE奥尼罗非鱼的总胆固醇(total cholesterol, TC)在感染后3 d显著降低(P<0.05)。血液生化指标表明感染无乳链球菌会引起4种奥尼罗非鱼的肝、肾损伤,肌肉受损和脂代谢障碍,但EX奥尼罗非鱼的肝和肌肉受损最小,EX和EE奥尼罗非鱼代谢活动影响较小。上述结果表明,4种奥尼鱼中EX奥尼罗非鱼的耐链球菌病能力最强。本研究从耐链球菌病能力角度评价了4种不同罗非鱼正交组合间的合配效果和育种潜力,预期结果为培育罗非鱼抗链球菌病新品种提供理论依据和育种材料。
Disease outbreak from Streptococcus agalactiae infection has become the most serious bacterial disease for tilapia(Oreochromis spp.) aquaculture in China, while effective prevention and control methods have not yet been established. It is an important approach to breeding the most S. agalactiae-resistant hybrid tilapia for better control and prevention S. agalactiae in tilapia. 4 family groups of hybrid tilapia F1 with different parents were got. They were O. niloticus Egypt strain ♀×O. aureus Xia'ao strain ♂(NE ♀×AX ♂, EX),O. niloticus Xianghu strain ♀×O. aureus Xia'ao strain ♂(NX ♀×AX ♂, XX), O. niloticus Xianghu strain ♀×O.aureus Egypt strain ♂(NX ♀×AE ♂, XE), O. niloticus Egypt strain ♀×O. aureus Egypt strain ♂(NE ♀×AE ♂,EE). The offspring were strengthened rearing for 100 d in the ponds and taken back to the laboratory. All fish were acclimated for 14 d in the laboratory then artificially infected by S. agalactiae intraperitoneally. Fish were cultured on 33 ± 1 ℃. The relative survival rate and serum biochemical parameters of 4 hybrid tilapia infected by S. agalactiae were investigated. The relative survival rate was recorded at different times post infection; Blood samples were respectively collected and analyzed. The relative survival rate at 7 d after infection was as follows: EX hybrid tilapia(50%) > EE hybrid tilapia(37.5%) > XE hybrid tilapia(33.3%)>XX hybrid tilapia(20.8%). Alanine transaminase(ALT) of EX hybrid tilapia showed no significantly difference(P>0.05) before and post injection, while that of other 3 species was significantly higher after infection(P<0.05);The serum electrolytes of 4 hybrid tilapia showed significantly difference after infection by S. agalactiae(P<0.05), but Na+, Ca2+ and Cl-of EX hybrid tilapia almost returned to control level at 3 d after infection. Globulin(GLO) of 4 hybrid tilapia increased statistically at 3 d after infection, and peaked at 7d, while the albumin/globulin(A/G) index of 4 hybrid tilapia declined significantly at the time(P<0.05);Lactic dehydrogenase(LDH) of all 4 hybrid tilapia increased significantly on the progress of disease(P<0.05), and peaked with approximately 1.81-fold, 6.1-fold, 2.42-fold and 3.77-fold when compared with control samples prior to infection, respectively. Superoxide dismutase(SOD) activity of EE hybrid tilapia was significantly lower than that of the other species prior to infection, but SOD activity of EX hybrid tilapia was the lowest at 1 d and 7 d after infection. The triglyceride(TG) of 4 hybrid tilapia decreased significantly at 3 d after infection by S. agalactiae(P<0.05), while the total cholesterol(TC) of EX, XX and XE decreased significantly at 3 d after infection(P<0.05). Biochemical indexes had shown the 4 hybrid tilapia manifested liver and kidney damage, muscle injury and lipid metabolism after infection, while EX hybrid tilapia diserved less liver damage and muscle injury. And EX and EE hybrid tilapia showed less injury on metabolic activity.These results indicated that EX hybrid tilapia was the most resistant to S. agalactiae. In this study, the potential effects of the different tilapia orthogonal combinations were evaluated from the perspective of antistreptococcosis. The expected results provide theoretical basis and potentially effective for genetic improvement of tilapia S. agalactiae-resistant breeding.
Disease outbreak from Streptococcus agalactiae infection has become the most serious bacterial disease for tilapia(Oreochromis spp.) aquaculture in China, while effective prevention and control methods have not yet been established. It is an important approach to breeding the most S. agalactiae-resistant hybrid tilapia for better control and prevention S. agalactiae in tilapia. 4 family groups of hybrid tilapia F1 with different parents were got. They were O. niloticus Egypt strain ♀×O. aureus Xia'ao strain ♂(NE ♀×AX ♂, EX),O. niloticus Xianghu strain ♀×O. aureus Xia'ao strain ♂(NX ♀×AX ♂, XX), O. niloticus Xianghu strain ♀×O.aureus Egypt strain ♂(NX ♀×AE ♂, XE), O. niloticus Egypt strain ♀×O. aureus Egypt strain ♂(NE ♀×AE ♂,EE). The offspring were strengthened rearing for 100 d in the ponds and taken back to the laboratory. All fish were acclimated for 14 d in the laboratory then artificially infected by S. agalactiae intraperitoneally. Fish were cultured on 33 ± 1 ℃. The relative survival rate and serum biochemical parameters of 4 hybrid tilapia infected by S. agalactiae were investigated. The relative survival rate was recorded at different times post infection; Blood samples were respectively collected and analyzed. The relative survival rate at 7 d after infection was as follows: EX hybrid tilapia(50%) > EE hybrid tilapia(37.5%) > XE hybrid tilapia(33.3%)>XX hybrid tilapia(20.8%). Alanine transaminase(ALT) of EX hybrid tilapia showed no significantly difference(P>0.05) before and post injection, while that of other 3 species was significantly higher after infection(P<0.05);The serum electrolytes of 4 hybrid tilapia showed significantly difference after infection by S. agalactiae(P<0.05), but Na+, Ca2+ and Cl-of EX hybrid tilapia almost returned to control level at 3 d after infection. Globulin(GLO) of 4 hybrid tilapia increased statistically at 3 d after infection, and peaked at 7d, while the albumin/globulin(A/G) index of 4 hybrid tilapia declined significantly at the time(P<0.05);Lactic dehydrogenase(LDH) of all 4 hybrid tilapia increased significantly on the progress of disease(P<0.05), and peaked with approximately 1.81-fold, 6.1-fold, 2.42-fold and 3.77-fold when compared with control samples prior to infection, respectively. Superoxide dismutase(SOD) activity of EE hybrid tilapia was significantly lower than that of the other species prior to infection, but SOD activity of EX hybrid tilapia was the lowest at 1 d and 7 d after infection. The triglyceride(TG) of 4 hybrid tilapia decreased significantly at 3 d after infection by S. agalactiae(P<0.05), while the total cholesterol(TC) of EX, XX and XE decreased significantly at 3 d after infection(P<0.05). Biochemical indexes had shown the 4 hybrid tilapia manifested liver and kidney damage, muscle injury and lipid metabolism after infection, while EX hybrid tilapia diserved less liver damage and muscle injury. And EX and EE hybrid tilapia showed less injury on metabolic activity.These results indicated that EX hybrid tilapia was the most resistant to S. agalactiae. In this study, the potential effects of the different tilapia orthogonal combinations were evaluated from the perspective of antistreptococcosis. The expected results provide theoretical basis and potentially effective for genetic improvement of tilapia S. agalactiae-resistant breeding.
引文
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金珊,王国良,赵青松,等.2005.加州鲈白云病的病原及血液病理的初步研究[J].水生生物学报,29(2):184-188.(Jin S,Wang G L,Zhao Q S,et al.2005.Preliminary studies on pathogen and hemopathology of the white cloud disease in MicroPterus salmoides[J].Acta Hydrobiologica Sinica,29(2):184-188.)
李思发,颜标,蔡完其,等.2008.尼罗罗非鱼与萨罗罗非鱼正反杂交后代耐盐性能的杂种优势及其与遗传的相关性的SSR分析[J].中国水产科学,15(2):189-197.(Li S FYan B,Cai W Q,et al.2008.Heterosis and related genetic analysis by SSR for the salt tolerance of reciplrocal hybrids between Nile tilapia(Oreochromis niloticus)and blackchin tilapia(Sarotherodon melanotheron)[J].Journal of Fishery Sciences of China,15(2):189-197.)
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