广西德保猪抵抗素基因的克隆与分析

[目的]克隆广西德保猪抵抗素（Resistin,RENT）基因并进行系统生物信息学分析,阐明德保猪脂肪细胞分化与脂肪生成的分子机制.[方法]根据NCBI已公布的猪RENT基因序列（NM 213783.2）设计特异性引物,以从德保猪血液中提取的总RNA为模板,PCR扩增RENT基因序列,T-A克隆至pEASY-T1载体后进行测序分析,并将所得测序结果用BLAST程序、MEGA 5.0软件、EXPASY服务器、SMART程序、SignIP程序、Softberry服务器和DNASTAR软件进行生物信息学分析.[结果]扩增获得468 bp的RENT基因序列片段,包含全长的德保猪RENT基因编码区和部分非编码区序列.多重序列比较分析结果显示,德保猪RENT基因编码区核酸序列与猪、牛、山羊、绵羊、人、兔、猕猴和豚鼠的同源性分别为100.0％、85.5％、85.5％、84.5％、81.2％、80.3％、80.0％和77.3％.德保猪RENT蛋白结构预测结果显示,其理论分子质量11.69 ku,等电点7.82,为弱碱性蛋白;N-末端存在信号肽,定位于胞质外,仅具有1个低复杂度结构域,包含1个α-螺旋、8个β-折叠、7个T-转角及两个无规则卷曲.[结论]RENT基因在德保猪中呈高度保守特征,是其脂肪细胞分化中的重要调控基因.     

Aeromonas sobria, a causative agent of disease in farmed perch, Perca fluviatilis L

Significant numbers of perch, Perca fluviatilis, raised on a pilot fish farm in Switzerland presented focal skin lesions on the lateral sides and fin rot. Mortality rates reached levels of up to 1% of the total fish on the farm per day. Virtually pure cultures of Aeromonas sobria were isolated from the liver, kidney, spleen and skin lesions of affected fish. Aeromonas sobria isolated from the farmed perch had a haemolytic effect on sheep and trout erythrocytes, autoaggregated, was cytotoxic for cultured fish cells and possessed genes involved in type III protein secretion. Experimental infection of naive perch with a single colony isolate of A. sobria from an affected farm fish resulted in the development of clinical signs identical to those seen on the farm. The results indicate that A. sobria can act as a primary pathogen of perch.     

Effects of High Pressure on Myofibrillar Protein and Moisture Distribution of Shrimp (Solenocera melantho) Muscle

ABSTRACT

The effects of high pressure (100–500 MPa) treatment on myofibrillar protein (MP) contents and distribution of water in Solenocera melantho muscle were investigated. The results showed that the shrimp treated with 200 MPa had the highest shelling score, and the solubility of MP in meat of shrimp treated with 300 MPa was significantly reduced. Moreover, the surface hydrophobicity of MP increased with increasing pressure. Treatment of shrimp with 200–300 MPa high pressure partially converted the structure of MP from an α-helix structure to a β-sheet structure, allowing for greater exposure of the tryptophan residues present. Treatment of shrimp with 100–200 MPa led to a significant loss in the amount of immobilized water in shrimp muscle. However, the treatment did not significantly change the moisture distribution. Overall, pressures of 100 to 200 MPa are considered the most suitable for the processing of S. melantho.     

Effect of dietary protein levels on growth of Penaeus stylirostris juveniles

The shrimp Penaeus stylirostris is currently produced on a commercial scale in Tahiti and New Caledonia. Both super-intensive (80 animals m⁻²) and semi intensive (25 m⁻²) systems are promoted. Locally produced commercial feed contains 380–400 g kg⁻¹ crude protein (CP) without special consideration for environmental impact. The need for a 'low pollution' diet implies reconsideration of the optimum dietary protein level for this species. Under experimental conditions, six isoenergetic practical diets ranging from 270 g kg⁻¹ to 440 g kg⁻¹ CP were formulated and fed to satiation for 30 days to juvenile P. stylirostris ; average growth rates were between 5.5 and 7.5 g per month with survival rates > 90%.
The lowest protein levels 270–310 g kg⁻¹, gave significantly ( P < 0.05) poorer growth (5.5 g per 30 days) than was observed with 330–430 g kg⁻¹ CP; 330 g kg⁻¹ CP may be recommended, and as it is lower than levels in diets used currently, there is a possibility of reducing nitrogenous waste. In addition to growth response, protein efficiency ratio, protein productive value and food conversion (feed/gain) all supported a recommended dietary protein level of 330 g kg⁻¹. Future prospects for practical feeds with even lower CP levels are considered.     

Optimal dietary protein requirement of grouper Epinephelus coioides juveniles fed isoenergetic diets in floating net cages

An experiment to determine the optimal protein requirement of grouper Epinephelus coioides juveniles was conducted in floating net cages (1.5 m × 1 m × 1.5 m). Six isoenergetic fishmeal–casein‐based experimental diets containing 350–600 g kg^?1 crude protein (CP) were fed to triplicate groups of 20 fish (10.7 ± 0.2 g) for 56 days. Weight gain (WG) and specific growth rate (SGR) increased with increasing dietary protein level from 350 to 450 g kg^?1 and then plateaued above these levels. Feed intake (FI) showed no significant difference among fish fed more than 350 g kg^?1 CP. Lowest feed conversion ratio (FCR) was found for fish fed 500 g kg^?1 CP but this was not significantly different from that of fish fed the 450 and 600 g kg^?1 CP. Lowest protein efficiency ratio (PER) was found for fish fed 550 and 600 g kg^?1 CP. Fish fed the 600 g kg^?1 CP had the highest body protein and moisture contents but the lowest body lipid content. Body ash content was unaffected by protein level for fish fed >400 g kg^?1 CP. Dietary protein level had no significant effect on hepatosomatic index (HSI). Fish fed the 350 g kg^?1 CP had significantly lower condition factor (CF) and viscerosomatic index (VSI). Based on broken‐line regression analysis of SGR the optimal dietary protein requirement for E. coioides juveniles was determined to be close to 480 g kg^?1.     

Dietary protein level and natural food management in the culture of blue (Litopenaeus stylirostris) and white shrimp (Litopenaeus vannamei) in microcosms

The effect of dietary protein level and natural food management on the production parameters of blue and white shrimp, as well as on water quality, was evaluated in a microcosms system (plastic pools simulating aquaculture ponds). Two experimental trials were carried out in the facilities of DICTUS, University of Sonora, Northwest México. Treatment with low protein diet (LP) consisted of a low protein input (diet with 250 g kg^?1 crude protein) through the culture period; treatment with high protein diet (HP) consisted of a high protein input (diet with 400 g kg^?1 crude protein) through the trial, and finally treatment VP consisted of an adjustment of protein input (diets with 250, 350 or 400 g kg^?1 crude protein), depending on the abundance of biota (zooplankton and benthos) in the system. Each species responded differently to the treatments. For blue shrimp, low protein input resulted in the lowest final body weight (12.9 ± 0.6 g) and biomass (696.0 g pool^?1). Survival and feed conversion ratio were similar in the three treatments. For white shrimp, the best growth, biomass and food conversion ratio were obtained in the low protein input treatment. Water quality parameters such as nitrate, ammonia and organic matter during the two trials, were better for LP and VP treatments. White shrimp seems to have lower protein requirements than blue shrimp. For the blue shrimp culture, adjusting protein input according to natural food abundance (zooplankton and benthos) in the system, seems to be advantageous because of the possibility of getting a production similar to that obtained with a high protein input through the farming period, but at lower feed cost, and with a lower environmental impact. It is concluded that a high protein input through the whole farming period is not the best feeding strategy for any of the two species.     

Effect of feeding level on growth and nutrient deposition in rainbow trout (Oncorhynchus mykiss Walbaum) growing from 150 to 600 g

The effects of feeding level on growth and energy partitioning were studied in rainbow trout growing from 150 to 600 g. Triplicate groups of fish (initial weight 158 g fish^?1) were fed a practical diet at various feeding levels (25%, 50%, 75% and 100% of near satiation) for 24 weeks at 8.5°C. The final body weights of fish were 235, 381, 526 and 621 g. Restricted feeding levels significantly reduced live weight gain. Feeding levels had less pronounced effects on feed efficiency ratio, which were 0.98, 1.08, 1.02 and 0.83, respectively, for the 25%, 50%, 75% and 100% feeding levels. The growth of fish fed to near satiation was accurately described by the thermal‐unit growth coefficient. The growth data also showed that the widely used specific growth rate was not an appropriate model. Fish fed at the lowest feeding level (25%), which represented a maintenance ration (energy gain was less than 2 kJ fish^?1 day^?1), showed positive protein deposition but negative lipid deposition. This indicates that fish fed a maintenance ration mobilize body lipid reserve to support protein deposition. The efficiency of energy for growth (k_g) was estimated to be 0.63. The factorial multiple regression approach estimated that the partial efficiencies of metabolizable energy utilization for protein deposition (k_p) and lipid deposition (k_f) were 0.63 and 0.72, and that maintenance energy requirement was about 19 kJ (kg BW^0.824)^?1, for rainbow trout held at 8.5°C.     

14C-glucose injection in Atlantic cod, Gadus morhua, metabolic responses and excretion via the gill membrane

In the present study cod were separately placed in fish metabolic chambers. Ten individuals were injected with a dose of 1 μCi ¹⁴C-glucose, in addition to 1 g pure glucose per kg live weight, and 10 individuals were kept under the same conditions and sham injected with 9 g L⁻¹ (0.9%) sterile saline solution.
Of the injected ¹⁴C-glucose, 3% was recovered in plasma, 2.9% in liver, 1.7% in red muscle, 18.2% in white muscle, 0.2% in heart, 1.9% in gills and 12.1% in gill-surrounding water. Of the 12.1% in gill-surrounding water, 9.1% was found to be in the form of CO₂, leaving 3% to pure glucose and lactate. These results indicate a relatively high production of CO₂ from glucose showing a utilization for energy, and a possible route of excretion of excess glucose via the gill membrane.
Activity of ¹⁴C-glucose on a concentration basis showed in descending order the following organs to be metabolically active in glucose utilization: gills > plasma (as a transporter) > heart > red muscle > liver > white muscle.
Of the total amount of ¹⁴C-glucose injected, only 0.3% was recovered in the liver lipid fraction, and mainly as triacylglyceroles; minor ¹⁴C-activity was found in mono- and diacylglyceroles and free fatty acids. These results show very low activity of de novo syntheses of lipid from injected glucose in the cod.     

Protein metabolism during sexual maturation in female Atlantic salmon (Salmo salar L)

Body composition and fractional rates of protein synthesis (percentage of the protein mass synthesized per day) were determined in female Atlantic salmon returning to the River Tay, Scotland in July and in October after a 95 day period without food, during which time the animals became sexually mature. During the 95 day period of starvation/sexual maturation the ventricle and red muscle remained as a constant proportion of fresh weight whereas the liver, gill and ovary increased and the stomach and white muscle decreased. Fractional rates of protein synthesis increased markedly in the liver, stomach and ovary during the period of starvation/sexual maturation. In the gill, ventricle and white muscle fractional protein synthesis rates increased slightly or remained constant. From the estimated rates of protein loss or gain in the various tissues it is concluded that there is considerable protein turnover and repartitioning of amino acids during the period of starvation and sexual maturation. The absolute rate of protein synthesis rates in the ovary indicates that this tissue made the largest contribution to the energy and amino acid demands of the fish, whilst most of the amino acids required for maturation of the ovary were derived from white muscle, principally as the result of increased muscle protein degradation.     

A comparison of RNA concentrations and ornithine decarboxylase activity in Atlantic salmon (Salmo salar) muscle tissue,with respect to specific growth rates and diel variations

RNA concentrations and enzyme activities are often used as indices of recent growth in fish, but few studies have used both methods to assess the same fish. This study measured RNA concentrations and ornithine decarboxylase (ODC) activity in muscle tissue of juvenile Atlantic salmon (Salmo salar) to compare their usefulness for reflecting specific growth rates, and to determine whether either growth index was influenced by diel variations or time of feeding. Three groups (n = 54 in total) were fed 1.5% of body weight in commercial pellets in four feedings per day. One group was fed only in the morning (0830–1230h), one in the afternoon (1430–1830h), and one in the morning and afternoon (0830–1830h). At the end of ten days, fish were sampled at three times (0130h, 1030h, 1630h) over a single 24h period. Correlations to specific growth rate were slightly higher for RNA concentrations than for ODC activity, but both were highly significant. RNA and ODC activity were also correlated to each other. These results suggest that RNA concentration and ODC activity, taken together, can be used to monitor changes in both the numbers and activity of ribosomes. For RNA concentrations, there was no evidence of an effect of diel variations or the time of feeding. For ODC activity, a significant diel effect (all feed schedules combined) was detected if one non-growing fish was excluded from the analysis; activity of the enzyme was slightly higher in the sample taken at night (0130h) than in the two daytime samples. Contribution no. 8, Catamaran Brook Habitat Research Project