不同抗性茭白感染纹枯病菌后4种酶活性的变化

选择3个对纹枯病抗性不同的茭白品种,研究其生物酶活性与抗性的关系。结果表明：抗性强的品种过氧化氢酶、多酚氧化酶和苯丙氨酸解氨酶的活性高于抗性弱的品种;人工接种以后,各品种间过氧化物酶、多酚氧化酶和苯丙氨酸解氨酶的活性都有不同程度的提高,抗性强的品种峰值高,抗性弱的品种苯丙氨酸解氨酶的活性增幅较大。     

响应曲面法分析低盐条件下温度与pH对斜带石斑鱼幼鱼能量代谢的联合效应

采用二因素五水平的中心复合设计和响应曲面分析方法,以温度(21~33℃)和pH(6.5~8.5)为因子,以耗氧率和排氨率为响应指标,探讨了在低盐条件下温度和pH对斜带石斑鱼(Epinephelus coioides)幼鱼能量代谢的联合影响。结果表明,本实验条件下,随着温度的上升,耗氧率(MO_2)和排氨率(MTAN)呈上升的变化;随着pH的上升,MO_2和MTAN呈先上升后下降的变化。温度一次效应对幼鱼的MO_2影响极显著(P0.01),二次效应对幼鱼的MO_2影响显著(P0.05);pH一次效应对幼鱼MO_2影响显著(P0.05),二次效应影响极显著(P0.01);温度与pH互作效应对MO_2影响显著(P0.05)。温度一次效应对幼鱼的MTAN影响极显著(P0.01),二次效应对幼鱼的MTAN无显著影响(P0.05);pH一次效应和二次效应对幼鱼MTAN影响显著(P0.05);温度与pH互作效应对MTAN无显著影响(P0.05)。所考察因子与幼鱼MO_2、MTAN间二次多项回归过程的决定系数分别为0.981 1、0.954 6(P0.01),可用于分析与预测。通过对所建立的回归方程同时优化,得到温度与pH的最佳组合为:温度31.2℃和pH 7.6。在此组合条件下,幼鱼MO_2、MTAN最大值分别为188.44 mg O_2·(kg·hr)-1;7.01 mg TAN·(kg·hr)-1,可靠度为0.902。实验结果提示,低盐条件下斜带石斑鱼养殖可在本实验中得到的最优因子组合下进行,以提高生产效益。     

氨氮和亚硝基氮共同胁迫对凡纳滨对虾感染WSSV的影响

为了评价养殖水环境中氨氮和亚硝基氮对凡纳滨对虾(Litopenaeus vannamei)的危害性,开展了氨氮和亚硝基氮共同胁迫对凡纳滨对虾感染WSSV后的死亡率、WSSV在患病对虾体内增殖速率和对虾主要免疫相关酶活性影响的研究。实验设置氨氮(NH+4)和亚硝基氮(NO-2)的共同胁迫浓度均为20 mg·L-1,分别注射10-4和10-5稀释度的WSSV提取液。结果显示,胁迫下感染10-4WSSV的凡纳滨对虾144 h死亡率达到100%,显著高于无胁迫组(76.67%),相同实验条件下高浓度病毒感染组死亡率高于低浓度组。对虾鳃组织WSSV荧光定量PCR检测结果显示,氨氮和亚硝基氮共同胁迫下凡纳滨对虾体内WSSV的增殖加快,感染48 h后胁迫组病毒量是无胁迫组的1.6倍,72 h时病毒量达到无胁迫组的2.0~3.7倍。此外,免疫相关酶活性结果显示,氨氮和亚硝基氮浓度突变会促使对虾血清中酚氧化酶(PO)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)活性先短暂升高然后降低。由此可见,氨氮和亚硝基氮共同胁迫会加快WSSV在患病凡纳滨对虾体内的增殖,导致更高死亡率,这可能是因为胁迫造成了对虾免疫相关酶活性的降低和抗病原感染能力下降所致。     

Immune responses of Litopenaeus vannamei to non‐ionic ammonia stress: a comparative study on shrimps in freshwater and seawater conditions

To investigate the effect of non‐ionic ammonia (NH₃‐N) stress (0.1 and 0.5 mg L^?1) on the immunity of Litopenaeus vannamei cultured in long‐term freshwater, the total haemocyte count (THC), the activity of phenoloxidase (PO), nitric oxide synthase (NOS), superoxidase dismutase (SOD) and the content of malondialdehyde (MDA) were determined and further compared with those of seawater shrimps. The results showed that NH₃‐N stress significantly reduced THC and the activity of PO and SOD (P < 0.05). Under 0.1 mg L^?1 NH₃‐N stress, NOS activity increased first and then decreased significantly, while it dropped dramatically under 0.5 mg L^?1 NH₃‐N stress (P < 0.05). During NH₃‐N stress, MDA content increased continuously, and the MDA content in hepatopancreas of freshwater shrimps was higher than that of seawater shrimps. It was concluded that NH₃‐N stress significantly influenced the non‐specific immunity and could also upset the balance of antioxidant system of L. vannamei in both freshwater and seawater shrimps. Compared with in seawater, the shrimps in freshwater were more vulnerable to NH₃‐N stress because of higher lipid peroxidation and lower immunity.     

Ammonia nitrogen excretion in Mandarin Fish (Siniperca chuatsi) and Grass Carp (Ctenopharyngodon idellus) fed practical diets: the effects of water temperature

To understand the actual production of fish culture about the utilization of dietary protein and excreta impact on the environment between mandarin fish (Siniperca chuatsi) and grass carp (Ctenopharyngodon idellus), the study to investigate the effect of temperatures (19 ± 0.5°C, 24 ± 0.5°C and 29 ± 0.5°C) on ammonia‐N excretion in mandarin fish and grass carp under fed and fasted states was conducted. These two species were fed a practical diet containing 325.2 g kg^?1 crude protein at 3% body weight per day. The ammonia‐N excretion rate was significantly increased when temperature increased from 19 to 29°C, and a linear relationship between ammonia‐N excretion rate and temperature. The maximum ammonia‐N excretion levels of mandarin fish and grass carp were observed at 4–8 h and 2–4 h after feeding, respectively, and the minimum values for both species were observed at 24 h after feeding. Under the feeding condition, mandarin fish had a lower ammonia‐N excretion level compared to grass carp at 24°C and 29°C. The average amount of ammonia‐N excreted by mandarin fish at 24 h is significantly higher than grass carp under fasting conditions, except 19 ± 0.5°C. These results indicated that mandarin fish might make better use of protein at higher temperature than grass carp when fed practical diets in commercial production. These results of this study suggested that mandarin fish had a lower ammonia‐N excretion level compared with grass carp, making a less contribution to environmental loading in an intensive fish culture.     

赤眼鳟耗氧率、排氨率和窒息点的初步研究

采取封闭流水式的实验方法,研究了对不同温度(16℃、20℃、24℃、28℃)对体质量小规格(6.98±1.19)g和大规格(22.45±1.79)g的赤眼鳟(Squaliobarbus curriculus)幼鱼耗氧率、排氨率以及窒息点的影响。结果表明,温度(X)对小规格和大规格赤眼鳟的耗氧率(Yo)和排氨率(YN)有显著性影响,其两两间的相关关系可分别用方程式:小规格赤眼鳟Y_o=0.012x-0.135(R~2=0.969),大规格赤眼鳟Y_o=0.007x-0.058(R~2=0.991);小规格赤眼鳟Y_N=0.001x-0.010(R~2=0.981),大规格赤眼鳟Y_N=0.001e~(0.085 x)(R~2=0.943)来表示。在16~28℃范围内赤眼鳟幼鱼的耗氧率和排氨率随着温度的升高而呈显著上升。在16℃时赤眼鳟的排氨率要显著性的低于20~28℃,而在20~28℃之间赤眼鳟幼鱼的排氨率差异性不显著。耗氧率昼夜变化表明大、小规格赤眼鳟白天平均耗氧率高于夜间。16℃条件下,小规格和大规格赤眼鳟的窒息点最低分别为0.860和0.960mg/L,显著性低于20~28℃。     

褐煤的碳缓释特征及其对池塘底泥脱氮作用的影响

厌氧氨氧化和反硝化作用是底泥生物脱氮的主要过程,碳源是调控厌氧氨氧化和反硝化作用的关键因子。本研究以褐煤为对象,对褐煤的静态碳释情况及其对池塘底泥中脱氮作用的影响进行了研究。结果显示,褐煤在室温条件下的碳释放规律符合二级动力学方程,具备作为反硝化碳源的可行性;在脱氮实验中,发现褐煤对底泥上覆水体中的亚硝酸盐氮(NNO₂^--N)的去除具有促进作用,NNO₂^--N的去除率随褐煤浓度的增加而升高,当褐煤质量浓度为40 g/L时,N\${\rm{O}}_2^ - $\-N去除率最高达99.61%,此时硝酸盐氮(NO₃^--N)的浓度也最低;同时发现,水体中氨氮(NH₄⁺-N)氧化的最适褐煤质量浓度为10 g/L,其去除率达99.39%;对底泥中的厌氧氨氧化菌群进行Illumina高通量测序发现,其中浮霉菌门占比最大(39.6%~71.8%),优势菌属为Candidatus Brocadia (13.9%~35.8%)和Desulfovibrio (17.1%~34.8%),添加褐煤组Candidatus Scalindua菌属比例高于未添加组;荧光定量PCR得出,随着褐煤质量浓度升高,底泥中的反硝化菌丰度呈增长趋势,而厌氧氨氧化菌丰度则低于无褐煤添加组,表明添加褐煤对底泥反硝化有促进作用,而对厌氧氨氧化有一定的抑制作用。研究表明,褐煤具备作为反硝化碳源的条件,可用于池塘养殖底泥脱氮作用。     

碱胁迫下罗非鱼鳃氨转运蛋白Rh基因的表达

为研究碱环境下广盐性鱼类氨转运途径,将尼罗罗非鱼同时进行急性碱度胁迫(2、4、6 g/L)和慢性碱度胁迫,检测胁迫后120 h内的血氨浓度与鳃组织中Rhag、Rhbg、Rhcg1和Rhcg2基因mRNA表达变化,并采用免疫组化技术观察鳃组织中Rh蛋白的阳性反应。结果表明,急性碱度胁迫下,血氨浓度在12 h内快速升高到达峰值,4种Rh基因表达量升高,并于24 h到达峰值;慢性碱胁迫组由于碱度不断升高,血氨浓度呈波动状态,4种Rh基因的表达量均维持在较高表达水平,表明几种Rh蛋白均可能参与血氨浓度调节。胁迫24 h,Rhcg1基因的表达量显著高于其他3种基因。免疫组化结果表明,急性与慢性胁迫组中Rhag、Rhbg和Rhcg在鳃组织中均发现阳性反应,且随碱度升高,阳性反应增强。本研究表明,在碱胁迫环境下,尼罗罗非鱼会增强Rh基因与蛋白表达量参与氨转运过程。     

不同生物絮团对脊尾白虾高密度养殖水体氨氮的影响

为筛选适宜虾类工厂化养殖使用的生物絮团种类,以脊尾白虾(Exopalaemon carinicauda)为实验材料,探讨了3种不同产地(河南、福建、河北)来源的EM菌产生的生物絮团对脊尾白虾高密度养殖水体氨氮(ammonia nitrogen, AN)浓度的影响。每种生物絮团下共设置600尾/m~3、800尾/m~3、1 000尾/m~3共3个养殖密度,实验周期为8 d。结果显示,应用河南产地的EM菌,在600尾/m~3、800尾/m~3、1 000尾/m~3养殖密度下,水体最终氨氮浓度为1.28 mg/L、1.52 mg/L、1.90 mg/L,日均节水率为50.1%;应用福建产地的EM菌,水体最终氨氮浓度为1.03 mg/L、1.48 mg/L、2.15 mg/L,日均节水率为52.2%;应用河北产地的EM菌,水体最终氨氮浓度为1.58 mg/L、1.78 mg/L、2.74 mg/L,日均节水率为24.4%;而对照组水体最终氨氮浓度分别为1.62 mg/L、2.12 mg/L、3.05 mg/L,以上3种生物絮团在脊尾白虾高密度海水养殖中均有降低水体氨氮的作用,且效果存在显著差异,揭示水产养殖过程中应对适宜的EM菌试剂进行筛选后使用。实验筛选获得了适合脊尾白虾高密度养殖的生物絮团,为进一步开展其工厂化养殖及节水减排提供了参考。     

TaqMan real-time PCR assay for relative quantification of white spot syndrome virus infection in Penaeus monodon Fabricius exposed to ammonia

White spot disease is caused by a highly virulent pathogen, the white spot syndrome virus (WSSV). The disease is usually triggered by changes in environmental parameters causing severe losses to the shrimp industry. This study was undertaken to quantify the relative WSSV load in shrimp exposed to ammonia, using a TaqMan‐based real‐time PCR, and their subsequent susceptibility to WSSV. Shrimp were exposed to different levels of total ammonia nitrogen (TAN) (8.1, 3.8 and 1.1 mg L^?1) for 10 days and challenged with WSSV by feeding WSSV‐positive shrimp. WSSV was detected simultaneously in haemolymph, gills and pereopods at four hours post‐infection. The TaqMan real‐time PCR assay showed a highly dynamic detection limit that spanned over 6 log₁₀ concentrations of DNA and high reproducibility (standard deviation 0.33–1.42) and small correlation of variability (CV) (1.89–3.85%). Shrimp exposed to ammonia had significantly higher (P < 0.01) WSSV load compared to the positive control, which was not exposed to ammonia. Shrimp exposed to 8.1 mg L^?1 of TAN had the highest (P < 0.01) WSSV load in all three organs in comparison with those exposed to 3.8 and 1.1 mg L^?1 of TAN. However, haemolymph had significantly higher (P < 0.01) viral load compared to the gills and pereopods. Results showed that shrimp exposed to ammonia levels as low as 1.1 mg L^?1 (TAN) had increased susceptibility to WSSV.