Impact of sublethal di‐n‐butyl phthalate on the aquaculture fish species Nile tilapia (Oreochromis niloticus): histopathology and oxidative stress assessment

Phthalates such as di‐n‐butyl phthalate (DBP) and their esters are widely used plasticizers, their ubiquitous presence in daily life, inevitably leads to their restricted use due to important environmental pollution and health impacts and endocrine disruption potential. The aim of this study was to examine the effects of a sublethal concentration of 10 mg L^?1 DBP on haematocrit (HCT) values, gills and liver histology, malondialdehyde (MDA, 2‐thiobarbituric acid‐TBA reactivity) and reduced glutathione (GSH) levels in gills and liver tissue as oxidative stress biomarkers in the aquaculture fish species Nile tilapia (Oreochromis niloticus) after 24 (DBP‐24) and 96 (DBP‐96) h exposure. No differences were found between per cent HCT values in the 24 h exposure groups (P > 0.05). Response of antioxidant defence systems in liver and gill tissues of the fish were dependent on exposure duration and changed to a higher extent during 96 h. MDA levels in liver tissue increased in DBP treated fish in comparison to the control fish. However, the differences between the exposure and control groups were not significant (P > 0.05). A statistically significant decrease (P > 0.05) was recorded in gill MDA levels in the DBP‐96 group when compared to the control and DBP‐24 groups. The liver GSH levels were unchanged in the DBP treated fish. However, GSH levels were significantly lower in the gill tissue of the DBP‐96 group. Exposure to DBP caused several degenerative changes in the histology of gill and liver tissue. Gills displayed hyperaemia, epithelial lifting, oedema, talengiectasia, epithelial hyperplasia and fusion of secondary lamellae, whereas in liver several circulatory anomalies (hyperaemia, blood congestion and sinosoid dilatation) and vacuolization of hepatocytes were observed. Histopathological results demonstrated that the gills were more affected than the liver perhaps due to their direct contact with DBP.     

3种品系尼罗罗非鱼(Oreochromis niloticus)盐碱耐受性和生长比较

为评估尼罗罗非鱼的耐盐碱性能,分别测定了上海、山东、河北3种品系尼罗罗非鱼鱼种96 h的半致死盐、碱度,并在不同盐碱混合浓度(S0A0、S10A0、S10A2、S10A4、S10A6)中进行为期60 d的养殖生长比较。单盐、单碱耐性研究表明,上海、山东、河北3种品系尼罗罗非鱼鱼种的96 h半致死盐度分别为18.528 g/L、20.347 g/L、19.342 g/L,96 h半致死Na HCO3碱度为8.827g/L、8.540 g/L、8.542 g/L。盐碱混合条件下,盐度为10时,96 h半致死碱度分别为河北品系(4.377g/L)上海品系(3.561 g/L)山东品系(3.108 g/L),品系之间差异显著(P0.05);盐度为15时,96 h半致死碱度分别为河北品系2.144 g/L,上海品系2.183 g/L,山东品系2.183 g/L,品系之间无显著差异(P0.05),高盐度下尼罗罗非鱼鱼种的碱度耐受性明显低于低盐度下的碱度耐受性。结果表明,尼罗罗非鱼日均增重率在S0A0、S10A0组间无显著差异(P0.05),随着盐碱浓度增加,盐碱S10A4和S10A6组中日均增重率呈下降趋势,河北品系表现出相对生长优势(P0.05)。研究结果为尼罗罗非鱼适宜养殖的盐碱范围的确定、耐盐碱品系的筛选提供了基础资料。     

Pansteatitis in polluted Olifants River impoundments: nutritional perspectives on fish in a eutrophic lake,Lake Loskop,South Africa

This study compares the aetiology of pansteatitis in Lake Loskop, relative to two other impoundments along the Olifants River. Macroscopic and microscopic pathology, age determination and analysis of stomach content, fatty acids and stable isotopes explain the high prevalence of pansteatitis in Oreochromis mossambicus (Peters) and several other species in Lake Loskop. All the dietary indicator comparisons between pansteatitis‐affected and healthy fish fail to support a systemic cause. Pansteatitis in Lake Loskop was linked to size and weight of O. mossambicus, but not to ontogenic age. Fish in Lake Loskop showed abnormally high omega‐3 to omega‐6 fatty acid ratios normally only found in marine fish with no significant difference in degree of assimilation of these fatty acids between pansteatitis‐affected and healthy fish. This explains the vulnerability to, but not the occurrence of, pansteatitis. As a cause for the pansteatitis, these results point towards sporadic vitamin E‐depleting trigger events, known sporadic fish die‐off occurrences that provide surviving fish with a rich source of rancid fats on which to scavenge. The mechanism ties pansteatitis to eutrophication and trophic cascade effects, the intrinsic drivers of the disease and suggests an adaptive management strategy that might be applied by relevant conservation authorities.     

尼罗罗非鱼表皮和鳃黏膜共生菌结构特征及其与鱼体健康状况相关关系

黏膜及其表面的共生菌群是鱼类抵御外界不利环境的第一道屏障。为探索养殖罗非鱼表皮和鳃黏膜共生菌菌群的结构特征是否与其健康状况间存在相关关系,本研究运用高通量测序技术,以无乳链球菌腹腔注射攻毒48 h后存活和濒死尼罗罗非鱼为检测对象,检测攻毒前后罗非鱼表皮和鳃黏膜共生菌群落结构差异。结果显示,健康尼罗罗非鱼表皮和鳃黏膜共生菌均存在优势菌群,主要为特吕珀菌属、硫杆菌属、弓形杆菌属、海单胞菌属和弧菌属。人工感染无乳链球菌后存活尼罗罗非鱼表皮和鳃黏膜共生菌菌群与感染前无显著差异;与存活组相比,濒死尼罗罗非鱼的表皮和鳃黏膜共生菌菌群多样性下降,其中弓形杆菌属、假交替单胞菌属、海单胞菌属、假单胞菌属和弧菌属等含量显著下降,链球菌属含量占总菌群的55.30%±1.24%,表明养殖尼罗罗非鱼表皮和鳃黏膜共生菌群落结构可能与其健康状况相关。     

无乳链球菌LIP蛋白的截短表达及免疫保护性研究

为评价罗非鱼源无乳链球菌(Streptococcus agalactiae)BX2012株脂蛋白(lipoprotein)的免疫原性及其对奥利亚罗非鱼(Oreochromis aureus)和大菱鲆(Scophthalmus maximus)的保护效果,以无乳链球菌脂蛋白基因序列(GenBank序列号:CP000114.1,SAK_0321)的B细胞线性抗原表位区设计特异性引物进行扩增,构建重组表达载体,将截短表达的脂蛋白制备成亚单位疫苗,同时制备灭活疫苗进行免疫比对。结果显示:重组表达载体p ET-32a-LIP342在BL21(DE3)中获得了良好的可溶性表达,分子质量约为30 kDa,纯化使LIP342纯度由41.75%提至87.23%;Western blotting分析显示,LIP342可被兔抗无乳链球菌高免血清特异性识别;LIP342在目前已知不同种属来源或不同血清型的无乳链球菌分离株中同源性为90.35%~100%,在罗非鱼源分离株中同源性为100%;无乳链球菌LIP342蛋白和灭活疫苗均可显著提升奥利亚罗非鱼和大菱鲆血清抗体水平,而LIP342诱导的血清抗体水平均显著高于灭活疫苗;经0.1 mL 1×10~9cfu/mL的无乳链球菌攻毒后,LIP342蛋白和灭活疫苗对供试鱼的累积存活率均显著高于PBS对照组。结果表明,高度保守的LIP342具有较好的免疫原性,可作为无乳链球菌亚单位疫苗候选因子。     

Detection of transgenic DNA in tilapias (Oreochromis niloticus, GIFT strain) fed genetically modified soybeans (Roundup Ready)

We used nested-polymerase chain reaction (PCR) to detect Roundup Ready soybean in aquatic feeds and feeding tilapias. A template concentration of 10⁻¹⁰ g μL⁻¹ DNA solution could be detected with a dilute degree of 0.01%. Most (90.6%) of the aquatic feeds containing soybean byproduct included exogenous DNA segments. We also compared genetically modified (GM) soybean with non-GM soybean diets in feeding tilapias ( Oreochromis niloticus , GIFT strain) and examined the residual fragments (254 bp) of GM soybeans. Tilapias receiving GM soybean diets had DNA fragments in different tissues and organs, indicating that exogenous GM genes were absorbed systemically and not completely degraded by the tilapia's alimentary canal.     

Re-evaluation of total sulphur amino acid requirement and determination of replacement value of cystine for methionine in semi-purified diets of juvenile Nile tilapia, Oreochromis niloticus

Two feeding experiments were conducted to re-evaluate the total sulphur amino acid (TSAA; methionine and cystine) requirement and determine the replacement value of cystine for methionine of juvenile Nile tilapia ( Oreochromis niloticus ). Semi-purified diets used in both experiments contained 3510 kcal gross energy and 280 g of protein per kilogram diet from casein, gelatin and crystalline amino acids. The basal diet of the first experiment contained 3.1 g methionine and 0.4 g cystine per kilogram. l -methionine was added to the seven remaining diets at 1.0 g kg⁻¹ increment to produce methionine levels ranging from 3.1 to 10.1 g kg⁻¹ diet. Each diet was fed to four replicate groups of juvenile Nile tilapia (1.28 g mean weight) in a recirculation system for 8 weeks. Broken-line regression analysis of weight gain data indicated that the TSAA requirement of juvenile Nile tilapia was 8.5 g kg⁻¹ of the diet or 30.4 g kg⁻¹ of dietary protein. In the second experiment, TSAA level was set at 95% of the requirement value determined in the first experiment. Seven diets were made with different ratios of l -methionine and l -cystine (20 : 80, 30 : 70, 40 : 60, 50 : 50, 60 : 40, 70 : 30 and 80 : 20, based on an equimolar sulphur basis). Each diet was also fed to four replicate groups of juvenile Nile tilapia (4.14 g mean weight) in a recirculation system for 8 weeks. Regression analysis of weight gain data using broken-line model indicated that cystine (on a molar sulphur basis) could replace up to 49% of methionine requirement in semi-purified diets for juvenile Nile tilapia.     

Effect of Bacillus cereus as a water or feed additive on the gut microbiota and immunological parameters of Nile tilapia

We evaluated the effects of Bacillus cereus, as an additive in water and feed, on the gut microbiota and immunological parameters of Nile tilapia (Oreochromis niloticus) fingerlings. Experiments were performed in tanks and net cages respectively. Experiment 1: Tilapia were housed in tanks for 42 days, and B. cereus was added to the water at 1.0 × 10⁴ cfu mL^?1 (Treatment 1) and 1.0 ×10⁵ cfu mL^?1 (Treatment 2) weekly. For the control, no probiotic was added. Experiment 2: Tilapia were housed in cages for 42 days, and the feed was supplemented with B. cereus at 1.0 × 10⁷ cfu g^?1 (Treatment 1) and 1.0 × 10⁸ cfu g^?1 (Treatment 2) weekly. For the control, no probiotic was added. Each treatment contained three replicates, with 50 male tilapias per replicate. The fish from the probiotic treatments in both tank and cage experiments had significantly higher serum lysozyme and peroxidase activities than the control. In the cage experiment, alkaline phosphatase and total superoxide dismutase activities in tilapia were significantly higher in probiotic treatments compared with the control. The results of polymerase chain reaction‐denaturing gradient gel electrophoresis showed that B. cereus supplementation in the feed and water affected the autochthonous gut bacteria community of tilapia and stimulated various potentially beneficial bacteria. Therefore, B. cereus, as a water or feed additive, could enhance the immune status and affect the gut microbiota of tilapia. Bacillus cereus was more effective as a feed supplement rather than a water additive for enhancing the immune status of tilapia.     

Effects of biofloc technology on reproduction and ovarian recrudescence in Nile tilapia

The objectives of this study were to evaluate the reproductive cycle, morphological changes of ovary and mobilization of energy reserves in Nile tilapia reared with biofloc technology (BFT). In general, the growth and reproductive performance were highly similar between BFT and Control system (clear water). Difference between the systems was found in the hepatosomatic index (using mixed‐effects models), which suggested that BFT can alter the energy mobilization in the post‐spawning period. The absolute and relative fecundity, fertilization rate, number of larvae produced per female, gonadosomatic index, proportion of oogenesis cells, number of post‐ovulatory and atretic follicles were similar between the two systems. We also did not detect a reduction in the reproductive cycle length in Nile tilapia reared in BFT. Because there was no evidence of the negative effects of BFT on Nile tilapia reproduction, we concluded that BFT might be used for breeder stocking of this species.     

盐酸半胱胺和二氢吡啶对奥利亚罗非鱼生长及相关激素水平、血液生化指标的影响

挑选大小相近的奥利亚罗非鱼270尾,随机分为3组,即对照组(基础日粮)、盐酸半胱胺(CHS)组(基础日粮 150 mg.kg-1CHS)、二氢吡啶(D il)组(基础日粮 100 mg.kg-1D il),试验期为32 d,分别于试验开始第1、15和32天测定体重、激素水平和生理生化指标的变化。结果表明,与对照组相比,CHS组、D il组雄鱼日增重分别提高40.3%、18.1%(P<0.05),特定生长率提高32.8%、16.7%(P<0.05),血中GH水平提高33.0%、11.6%,T3水平提高28.5%、40.4%,睾酮水平提高35.3%、28.7%;雌鱼日增重分别提高15.5%、25.7%,特定生长率提高8.7%、16.7%,GH水平提高34.4%、10.9%,T3水平提高28.1%、46.5%,睾酮水平提高35.3%、28.7%。CHS组和D il组雌、雄鱼脲氮水平明显下降(P<0.05);血清及肝脏抗氧化能力显著增强(P<0.05)。结果提示:CHS和D il均可促进奥利亚罗非鱼的生长,CHS对雄鱼促生长作用较强,D il对雌、雄鱼的促生长作用相似。