斑点叉尾鮰病毒囊膜蛋白ORF6在昆虫细胞中的表达

为构建基于杆状病毒表达系统的CCV新型亚单位疫苗,将CCV的囊膜蛋白基因ORF6克隆至杆状病毒转座载体pFastBacTM 1质粒中,并将阳性重组转座质粒转化进含穿梭载体Bacmid的感受态细胞DH10Bac中,获得重组子rBacmid-ORF6。在脂质体介导下将该重组子转染sf9昆虫细胞,获得重组杆状病毒AC-ORF6。AC-ORF6感染的sf9昆虫细胞,经超薄切片电镜观察,可见重组杆状病毒呈多粒包埋,经间接免疫荧光、Western-blotting检测,CCV的ORF6蛋白可以在感染了AC-ORF6的sf9细胞中表达。研究表明,获得了插入ORF6基因的重组杆状病毒,并且该基因可以在重组杆状病毒介导下在昆虫细胞中表达,从而为基于CCV ORF6的杆状病毒亚单位疫苗研究奠定了基础。     

Validation of a qPCR assay for the detection of Ictalurid herpesvirus‐2 (IcHV‐2) in fish tissues and cell culture supernatants

Ictalurid herpesvirus‐2 (IcHV‐2) is a pathogen of cultured black bullhead, Ameiurus melas (Rafinesque), and has been shown to produce high mortality in experimental exposures of channel catfish, Ictalurus punctatus (Rafinesque). During acute infections, the virus grows readily in cell cultures but produces a cytopathic effect (CPE) similar to that of Ictalurid herpesvirus‐1 (IcHV‐1) and the channel catfish reovirus. We have developed a quantitative PCR assay that can be used to detect IcHV‐2 in fish tissues and cell culture supernatants. The assay does not amplify other fish herpesviruses tested or host DNA. It is quantitative over a range of eight logs, and the limit of detection is <10 copies per reaction. In replicate assays carried out on different days, the coefficient of variability was 10%. The best organs for the detection of acute IcHV‐2 infections by our assay are the spleen and kidney. This assay should be useful for the diagnosis of IcHV‐2 disease, the identification of syncytial CPEs in cell cultures, and for the detection of latent infections in carrier fish.     

Evaluation of soybean meal in the formulated diets for juvenile Pangasianodon hypophthalmus (Sauvage, 1878)

The influence of soybean meal (SBM) on the growth and feed utilization of juvenile sutchi catfish (Pangasianodon hypophthalmus, Sauvage, 1878) was investigated. Eight isonitrogenous (300 g kg^?1 CP) and isoenergetic (18 MJ kg^?1) diets were formulated incorporating Argentine SBM to replace fish meal at 0, 150, 300, 450, 600, 750, 900 and 1000 g kg^?1 dietary protein. Each diet was fed to three replicate groups of fish with an initial weight 6.0–6.2 g for twelve weeks. Growth performance decreased, and feed utilization was worsened with the increase in SBM inclusion in the diets. Final weight and relative growth rate (RGR) of fish fed control diet (0 SBM) were significantly higher than those fed test diets (P < 0.05). However, there were no significant differences in the final weight and RGR between the fish fed on 15 SBM, 30 SBM, 45 SBM and 60 SBM diets. Specific growth rate of fish fed 0 SBM, 15 SBM, 30 SBM and 45 SBM diets was significantly higher than those fed other diets. Feed conversion ratio of fish fed 0 SBM, 15 SBM, 30 SBM, 45 SBM and 60 SBM diets was significantly lower than those fed other diets (P < 0.05). The dry matter and protein digestibility were lesser in all the diets in comparison with the control diet. Hepatosomatic index and viscerosomatic index increased with increasing SBM in diet. This present trial indicated that fish meal can be replaced by SBM in the diet of juvenile sutchi catfish only up to 45% of fish meal protein without any adverse effect on growth, feed utilization and body composition.     

Age‐0 blue catfish habitat use and population demographics in the middle Mississippi River

The demographics (e.g. hatch time, growth and mortality) and habitat (macrohabitat and mesohabitat) use of age‐0 blue catfish Ictalurus furcatus (Lesueur) were evaluated in the un‐impounded reach of the middle Mississippi River during 2003–2010. Each year, island complexes and adjacent main river habitats were trawled, and 7,373 age‐0 blue catfish (10–144 mm total length) were collected. Blue catfish hatch timing varied slightly across years and generally began in June and ended in August. Age‐0 blue catfish growth rates and instantaneous mortality rates differed among years. Catch rates of age‐0 blue catfish were highest in side channels and around islands, whereas main channel habitats were infrequently occupied. Regarding mesohabitat use, age‐0 blue catfish frequently occupied low velocity areas (i.e. 0.3–0.4 m/s) of shallow depth (i.e. 1–4 m) and sand substratum. This study has provided insight into age‐0 blue catfish population dynamics and has highlighted the importance of specific habitat characteristics in large rivers that may aid in recruitment of this species.     

Evaluation of potassium permanganate against an experimental subacute infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus (Rafinesque)

An experiment was performed to evaluate the efficacy of potassium permanganate (KMnO₄) as a prophylactic and therapeutic treatment of an experimental subacute infection of Flavobacterium columnare in channel catfish, Ictalurus punctatus . Fish were cutaneously abraded and divided into five treatment groups: (i) challenged by waterborne exposure to F. columnare and not treated with KMnO₄ (positive control), (ii) challenged and simultaneously treated with KMnO_4, (iii) challenged and treated with KMnO₄ at 1, 6 and 9 days post-challenge, (iv) not challenged and treated with KMnO₄ at 1, 6 and 9 days post-challenge (first negative control) and (v) not challenged and not treated (second negative control). The dosing of KMnO₄ was 2.0 mg L⁻¹ above the potassium permanganate demand for 2 h duration. The survival of the group challenged and simultaneously treated with KMnO₄ (99%) was significantly higher than the positive control (78%) and was not significantly different from the negative control groups. The challenged fish treated with KMnO₄ post-challenge had 7% higher survival than the positive control (85% compared with 78%), but that difference was not statistically significant. The results demonstrate that KMnO₄ has a clear prophylactic value but probably a marginal therapeutic value once the infection has established.     

黄颡鱼规模化养殖高产试验

黄颡鱼对生态环境的适应能力较强,通过驯化可摄食人工配合饲料。2004年笔者在河南省郑州市沟赵镇水产养殖场进行了黄颡鱼规模化养殖高产课题试验,取得了较高的经济效益,现将试验情况总结如下。1试验条件1.1池塘条件试验池塘共9口,每口池塘面积均为0.67 hm2,水深2.0~2.5 m,水源充足,水质符合渔业用水水质标准。每个池塘配备6 kW叶轮式增氧机1台。1.2池塘清整投放鱼种前约15 d以1500 kg/hm2的生石灰或375 kg/hm2的漂白粉清塘消毒,7 d后投施3000~4500 kg/hm2的有机肥料,待水体中大量出现浮游动物后投放鱼种。2试验方法2.1鱼种放养鱼种购自湖北…     

3种黄颡鱼SOCSs基因分子特性及致病菌胁迫下的表达模式

细胞因子信号传导抑制因子(SOCS)是JAK/STAT信号通路中细胞因子信号受体的重要抑制因子。为探究SOCS在鱼类免疫应答过程中的作用，本研究以杂交黄颡鱼“黄优1号”及其母本黄颡鱼和父本瓦氏黄颡鱼为研究对象，克隆了杂交黄颡鱼“黄优1号”3个免疫相关基因(SOCS1、SOCS2和SOCS3)，并对其生物信息学进行了比较分析；采用qRT-PCR技术研究了SOCS1、SOCS2和SOCS3基因分别在3种黄颡鱼感染嗜水气单胞菌和鮰爱德华氏菌后的表达模式。结果显示，杂交黄颡鱼“黄优1号”SOCS1、SOCS2和SOCS3基因的全长分别为561、735和672 bp，分别编码186、244和223个氨基酸，其预测的蛋白结构均含有保守的SH2结构域和SOCS盒。黄颡鱼、瓦氏黄颡鱼和杂交黄颡鱼“黄优1号”SOCS1、SOCS2和SOCS3基因的表达在被检的8个不同组织中呈现组织特异性。在两种致病菌感染后，杂交黄颡鱼“黄优1号”及其双亲的SOCS1、SOCS2和SOCS3基因在肝、鳃和头肾组织的表达量均在前24 h显著升高，随后逐渐恢复至正常水平。此外，鮰爱德华氏菌感染后，杂交黄颡鱼“黄优1号” SOCS1、SOCS2和SOCS3基因的表达量显著高于双亲，表现出较双亲更强的免疫调节能力且具有病原特异性。该结果为进一步解析黄颡鱼SOCS家族免疫防御调控机制提供了参考依据。     

基于小龙虾田间分布的稻虾共生机制探究

为探究稻虾共生机制，运用地笼捕获法研究了小龙虾分别在高频分布采样区（GP）、低频分布采样区（DP）和无虾对照区（CK）的田间分布特征，结合稻田土壤性状、病虫草害调查和稻谷产量性状分析，研究稻虾共作模式的生态效应。结果显示：小龙虾稻田分布数量与田面水深呈显著正相关关系，一元线性回归方程为y=197.69x-25.77；小龙虾高频分布可以改善土壤结构，显著降低0~10 cm土壤容重6.25%~11.76%，显著增加0~10 cm土层总毛管孔隙度8.08%~10.78%（P<0.05）；小龙虾高频分布显著增加0~10 cm土壤有机质含量6.24%~10.30%，显著增加0~20 cm土层的土壤微生物生物量碳（P<0.05），并能显著抑制杂草生长；小龙虾不同分布频度对病虫害和水稻产量及其构成因子无显著影响，但水稻产量随着分布频度的加大具有增加的趋势。因此，生产和科研上可适当区分小龙虾不同分布频度（水深）进行管理和研究，促进稻虾生态种养绿色可持续发展。     

稻虾综合种养对稻田土壤肥力和金属元素含量的影响

为研究稻虾综合种养对稻田土壤肥力和金属元素含量的影响,对潜江市稻虾综合种养田(渔沟以TRC表示、田面以PRC表示)和水稻单作田(以RM表示,作为对照)土壤不同土层(0～10 cm、10～20 cm和20～30 cm)的主要肥力指标和金属元素含量进行了对比。结果显示,各土层总氮、速效磷和总磷含量均表现出PRC RM TRC的趋势,有机质(20～30 cm土层除外)、氨氮含量表现出RM PRC TRC的趋势,硝态氮含量表现出TRC PRC RM的趋势。各土层的Cu、As、Mg和Mn含量均表现出TRC PRCRM的趋势,而Cd、Pb、Fe和Ca含量的变化趋势与之相反;除0～10 cm土层外,Cr、Zn和Ni含量在TRC、PRC和RM之间均无显著差异。根据《GB 15618—2018》中的污染风险筛选值,各土层的Cr、Cd、Pb、Zn、Ni、As含量均未超标,而Cu含量普遍超标,超标率的范围为14%～46%。研究表明,稻虾综合种养可能会促进田面土壤中氮磷的积累、降低渔沟土壤氮磷的含量,也可能引起土壤中Cu、As、Mg、Mn的积累及Cd、Pb、Fe和Ca的减少。     

稻田环境对鲤血清生化指标、肠道组织形态及细菌群落结构的影响

稻鱼共生(Rice-fish co-culture)作为“全球重要的农业遗产系统”，已成为我国发展可持续农业的重要模式。为研究稻田环境条件下，鲤血清生化特性、肠道显微结构及肠道微生物菌群多样性、物种丰度的影响。于2019年6月在连南瑶族自治县，分别选取稻田和池塘，放养规格大小一致的鲤幼鱼进行3个月的不投喂养殖。试验结束后统一进行采样与检测。结果表明：稻田环境显著提高了鲤的体长和体质量，显著降低了鲤血清谷丙转氨酶(ALT)活性和肠道微绒毛高度(P＜0.05)，血清补体C3和C4的含量有上升趋势，但差异不显著(P＞0.05)；在肠道微生物方面，稻田环境显著提高了鲤肠道内菌群多样性，并在门水平显著提高了疣微菌门(Verrucomicrobia)和变形菌门(Proteobacteria)的丰度(P＜0.05)，显著降低了梭杆菌门(Fusobacteria)的丰度(P＜0.05)；在纲水平，显著提高了α-变形菌纲(Alphaproteobacteria)和放线菌纲(Actinobacteria)的丰度；在属水平，显著降低鲸杆菌属(Cetobacterium)、邻单胞菌属(Plesiomonas)、拟杆菌属(Bacteroides)以及气单胞菌属(Aeromonas)的丰度(P＜0.05)。实验结果证实，稻田环境改变了肠道微生物多样性及与代谢能力有关的功能基因丰度，降低了部分条件致病菌的丰度，减少了疾病爆发的几率，同时有利于维持肝功能稳定。对于保持养殖鱼类健康生长有积极作用。