Live microbial feed supplement in aquaculture for improvement of stress tolerance

Two bacterial strains, Lactobacillus fructivorans, isolated from sea bream (Sparus aurata) gut, and Lactobacillus plantarum, isolated from human faeces, were administered simultaneously, during sea bream development, using Brachionus plicatilis and/or Artemia salina as vectors. The probiotic treatment significantly affected gut colonization. To test the probiotic influence on stress responsiveness, sea bream fry, 47 days post-hatching (p.h.), were subjected to pH stress (from 8.6 to 6.3) and cumulative mortality, cortisol levels and HSP70 gene expression were analysed. Cortisol was selected, since under stress conditions its level increases. HSP70 was selected with consideration of its wide involvement in response to a great number of injuries, and because it protects cells probably by binding and refolding damaged proteins. The results obtained indicated that the administration of probiotic to sea bream fry induced higher HSP70 levels, indicating a greater potentiality to respond to the harmful conditions possibly present in fish farms. This hypothesis is supported by the fact that the levels of cortisol found were significantly lower (P < 0.05) in both groups under probiotic treatment. When pH was used as a stressor, it induced a higher cumulative mortality in the control; the mortality was found to be significantly lower in both treated groups. Interestingly, a significant increase (P < 0.05) in HSP70 gene expression was observed in all stressed groups. These results suggest an improvement in tolerance to acute stress of fry fed with probiotics.     

Investigation of the nutritional requirements of Australian snapper Pagrus auratus (Bloch & Schneider 1801): digestibility of gelatinized wheat starch and clearance of an intra-peritoneal injection of d-glucose

Two experiments were performed to investigate the digestibility and utilization of carbohydrate sources by Australian snapper Pagrus auratus. In the first experiment, snapper of two different size classes (110 and 375 g) were fed a reference diet containing no starch (REF) or diets containing 150 (PN15), 250 (PN25), 350 (PN35) or 450 g kg^?1 (PN45) of 100% gelatinized wheat starch to investigate the interactive effects of fish size and starch inclusion level on apparent organic matter (OM) or gross energy (GE) digestibility (ADC), post‐prandial plasma glucose concentration, hepatosomatic index (HSI) and liver or tissue glycogen content. A second experiment used a 72 h time course study to investigate the ability of larger snapper (300–481 g) to clear an intra‐peritoneal injection of 1 g d ‐glucose kg^?1 body weight (BW). Organic matter and GE ADCs declined significantly in both fish sizes as the level of starch increased (PN45energy small fishenergy large fish). There was no interaction between fish size and inclusion level with respect to GE or OM ADCs. Gross energy ADC for both sized fish was described by the linear function GE ADC=104.97 (±3.39)–0.109 (±0.010) × inclusion level (R²=0.86). Hepatosomatic index, liver and muscle glycogen concentrations were significantly elevated in both small and large snapper‐fed diets containing gelatinized starch compared with snapper fed the REF diet. Three‐hour post‐prandial plasma glucose concentrations were not significantly affected by fish size, inclusion level or the interaction of these factors (REF=PN15=PN25=PN35=PN45), and ranged between 1.60 and 2.5 mM. The mean±SD resting level of plasma glucose (0 h) was 2.4±1.1 mM. Circulating levels of plasma glucose in snapper peaked at 18.9 mM approximately 3 h after intra‐peritoneal injection and fish exhibited hyperglycaemia for at least 12–18 h. There were no significant differences between the plasma glucose concentrations of snapper sampled 0, 18, 24, 48 or 72 h after injection (0=18=24=48=72<12< 1<3=6 h), indicating snapper required almost 18 h to regulate their circulating levels of glucose to near‐basal concentrations. Australian snapper are capable of digesting moderate levels of gelatinized wheat starch; however, increasing the dietary content of starch resulted in a reduction in OM and GE digestibility. Smaller snapper appear to be less capable of digesting gelatinized starch than larger fish, and levels above 250 and 350 g kg^?1 of diet are not recommended for small and large fish respectively. Snapper subjected to an intra‐peritoneal injection of d ‐glucose have prolonged hyperglycaemia; however, the post‐prandial response to the uptake of glucose from normally digested gelatinized starch appears to be more regulated.     

双向选择对团头鲂生化遗传变异的影响↑（*）

本文报导双向选择对团头鲂生化遗传变异的影响。试验材料鱼包括来自淤泥湖的原种，１９８６－１９９４年群体选育的第四代（“选育 Ｆ４”），以及１９９１、１９９４年连续兄妹杂交的第二代（“近交Ｆ２”）。计检测了８种同工酶，１９个位点。三群体的多态座位比例一致（５．２６％），但平均杂合度趋异。“选育Ｆ４”平均杂合度最高（０．０２１３），“近交Ｆ２”最低（０．０１７４）。经４代选育，平均杂合度增加４％，而经２代近交，平均杂合度降低１５％。     

Dietary eicosapentaenoic acid requirement of juvenile rock bream,Oplegnathus fasciatus

A feeding trial was conducted to evaluate the optimum dietary level of eicosapentaenoic acid (EPA, 20:5n‐3) based on growth and non‐specific immune responses in juvenile rock bream. A basal diet without EPA supplementation was used as a control, and six other diets were prepared by supplementing with 4, 8, 12, 16, 20 or 40 g of EPA per kg diet. The actual EPA concentrations of the diets were 0.5, 4.3, 8.5, 13.0, 16.8, 21.0 and 41.2 g of EPA per kg diet, and the diets were abbreviated as EPA_0.5, EPA_4.3, EPA_8.5, EPA_13.0, EPA_16.8, EPA_21.0 and EPA_41.2, respectively. Triplicate groups of fish averaging 1.06 ± 0.01 g (mean ± SD) were fed one of the seven experimental diets at the apparent satiation for 8 weeks. At the end of the feeding trial, weight gain, specific growth rate and feed efficiency of fish fed EPA_16.8, EPA_21.0 and EPA_41.2 diets were significantly higher than those of fish fed EPA_0.5, EPA_4.3, EPA_8.5 and EPA_13.0 diets (p < .05). Superoxide dismutase activity of fish fed EPA_16.8, EPA_21.0 and EPA_41.2 diets were significantly higher than those of fish fed EPA_0.5, EPA_4.3 and EPA_8.5 diets. Fish fed EPA_21.0 and EPA_41.2 diets showed significantly higher lysozyme activity than did fish fed EPA_0.5, EPA_4.3, EPA_8.5 and EPA_13.0 diets. The broken‐line analysis of weight gain indicated that the optimum dietary EPA level was 16.7 g/kg diet. These results suggested that the optimum dietary EPA level in juvenile rock bream could be greater than 16.7 g/kg diet but less than or equal to 16.8 g/kg diet based on the broken‐line analysis and the ANOVA test of weight gain.     

Growth of bream, Abramis brama (L.), in Irish canals and implications for management

The growth rates of bream, Abramis brama (L.), were determined in three Irish Canals: the Royal and Grand Canals, and the Barrow Line. Compared with growth patterns for other Irish and European waters, the growth of bream was fast in the Royal Canal, but slow to moderate for the Grand Canal and the Barrow Line. Significant differences in bream growth in terms of both length and weight were found between the three canal systems, and between canal levels, particularly in the Grand Canal. Associations between growth, and (1) alkalinity, (2) total fish biomass and (3) the ratio of bream biomass to total fish biomass were also found. However, it was considered that these three factors alone were not sufficient to explain the differences in growth. Productivity was implicated as the main factor controlling bream growth because fish species diversity was greater and total fish biomass was generally higher in levels of the Grand Canal with faster growing bream populations. The division between 'fast' and 'slow' growing bream also corresponded with a change in the aquatic weed community. There is evidence to suggest that there was a corresponding change in macro-invertebrate community structure and density. Angler usage as another factor in promoting growth is discussed. The implications for the future management and development of these canals are outlined.     

Studies on the green liver in cultured red sea bream fed low level and non-fish meal diets: Relationship between hepatic taurine and biliverdin levels

SUMMARY: The aim of this study was to investigate the cause of green liver in red sea bream fed substitute protein diets. Red sea bream Pagrus major was given either of the following diets for 28 weeks: (1) control diet (50% fish meal), (2) low level fish meal diet (15% fish meal), and (3) non-fish meal diet (0% fish meal). The green liver was observed in all groups tested, but the incidence was much higher in the experimental diet groups. The feeding of substitute protein diets reduced plasma triglyceride and cholesterol levels. However, there was no significant difference in plasma hepatic enzyme activities and plasma bile salts concentration among the treatments. Fish fed the substitute protein diets showed low hepatic taurine levels with an appearance of a biliverdin in the liver. Moreover, the proportion of ditaurobilirubin to total biliary bile pigments was significantly lower in fish fed the substitute protein diets. These data indicate that feeding of substitute protein diets did not induce any cholestatic hepatobiliary obstructions and that the low hepatic taurine level was one of the probable factors responsible for the occurrence of green liver in red sea bream fed substitute protein diets.     

Florfenicol depletion in edible tissue of rainbow trout,Oncorhynchus mykiss (Walbaum), and sea bream,Sparus aurata L.

An increase in fish production has consequently brought an increase in infectious diseases in fish farms. The use of chemotherapic drugs is the most effective instrument against common bacterial agents. The number of registered drugs for use in aquaculture is limited and often veterinary practitioners resort to the off‐label use of chemotherapic agents authorized for different food‐producing animal species. Florfenicol is well known for its outstanding effect against various pathogenic bacteria affecting fish, and therefore, it may be a useful drug for off‐label use in aquaculture. The aim of this study was to evaluate the depletion of florfenicol and its major metabolite, florfenicol amine, from the edible tissue of two fish species, rainbow trout and sea bream, following treatment with medicated feed at a dosage of 10 mg kg^?1 of bw day^?1, for 10 consecutive days. At prefixed time points after the end of administration (0.25, 1, 2, 3, 4, 6, 7, 10, 14 and 21 days after treatment), edible tissues (muscle plus adherent skin) from 15 individuals in each group were collected and analysed by HPLC, to determine concentration of the drug in the tissue. On the basis of the obtained concentrations, withdrawal times of florfenicol in the two species were calculated. The results indicate that a drug withdrawal time of 500 °C‐day, as established by Directive 2004/28/EC, for off‐label drug use is more than satisfactory to guarantee the healthiness of fish products against the risk of drug residues.     

不同倍性团头鲂群体遗传变异的初步分析

从鲤的微卫星DNA标记中筛选4对有效引物，进行不同倍性团头鲂遗传变异的微卫星DNA分析。结果表明：（1）4对引物中有2对能在不同倍性团头鲂五群体中探测到多态性，其等位基因数为4～6个，大小在131～307bp之间。发现两个等位基因（MFW19—175和MFW19—256）可作为异源3n特异的微卫星DNA标记。（2）不同倍性团头鲂五群体内的Shannon多样性指数为0．0562～0．2887，Nei’s基因多样性指数为0．0315～0．1969，平均遗传距离为0．0395～0．1542，不同倍性团头鲂群体存在较大的遗传变异。（3）反交3n、异源3n、正交3n和同源4n—F1群体的Shannon多样性指数和Nei’s基因多样性指数均显著地高于2n群体（P〈0．05）。（4）分子方差分析和聚类分析的结果表明：五群体分子系统树分成明显的两支，同源4n—F1、正交3n、反交3n和异源3n为一支，2n为另一支。     

Expression of recombinant triangular bream (Megalobrama terminalis) pre-IGF-I in Escherichia coli

The insulin-like growth factor I (IGF-I) cDNA (GenBank Accession No. AY247412) of triangular bream (Megalobrama terminalis) was expressed for the first time in Escherichia coli. To construct the expression plasmid, the IGF-I cDNA was subcloned into prokaryotic-expressing vector pGEX-4T-1. The E.coli JM109 was transformed with the recombinant plasmid pGEX-4T-1-IGF-I, and the transgene expression was observed after being induced with isopropyl-β-D-thiogalactoside (IPTG). The results of SDS polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting indicated that the recombinant fusion protein had immune activity and the molecular weight was about 47 kDa. The results of SDS-PAGE and thin layer scanning showed that the yield of fusion protein had been enlarged with prolonging time. When the time of induced expression was 1, 2, 3, 4, 5, and 6 h, the expression amount was approximately 1.4, 4.3, 8.1, 11.3, 16.3 and 18.8% of total bacterial protein respectively.     

Optimizing the Proportions of Plant Protein Ingredients in the Diet of Juvenile Blunt Snout Bream,Megalobrama amblycephala

An 8‐wk feeding trial was conducted to evaluate the influence of different proportions of dietary plant ingredients on the growth of juvenile blunt snout bream, Megalobrama amblycephala. A 2 × 3 factorial arrangement was set for this trial with three ratios of cottonseed meal (CSM) and canola meal (CM) (2:1, 1:1, or 1:2) and three proportions of soybean meal (SBM) (2, 1, or 0). The results showed that the specific growth rate and protein efficiency ratio both significantly decreased with decreasing amounts of SBM in the diets of blunt snout bream (P < 0.05), while the feed efficiency ratio and feed intake increased significantly (P < 0.05). Meanwhile, the growth performance and feed utilization were shown to improve with increasing amounts of CSM, although the effects were not significant (P > 0.05). The interaction of these two factors had no effect on either growth or feed utilization (P > 0.05). Diet composition was not seen to affect body composition (P > 0.05). The ratio of CSM to CM significantly affected gut amylase activity, liver alanine aminotransferase content, and plasma cholesterol levels (P < 0.05). The interaction between the two main effects significantly influenced the plasma glucose level (P < 0.05). Therefore, the suitable proportion of plant feed for blunt snout bream should include high levels of SBM and CSM, with a SBM : CSM : CM ratio of 2:2:1.