东方粘虫中肠V-ATPase H亚基原核表达及纯化

对东方粘虫Mythimna separate(Walker)中肠V-ATPase H亚基基因(VATPH)进行克隆、原核表达及纯化。首先采用RT-PCR技术克隆基因,再构建原核表达载体(pET15b-VATPH)并转入E.coil BL21(DE3),经异丙基硫代-β-D-半乳糖苷(isopropyl-β-D-thiogalactoside,IPTG)诱导表达,用Ni-NTA柱及S-200分子筛纯化,最后进行SDS-PAGE分析。结果表明,pET15b-VATPH可高效表达V-ATPase H亚基,纯化后可获得单一条带且分子质量约为55ku的重组蛋白。该结果为进一步研究V-ATPase H亚基的晶体结构,药物小分子与H亚基大分子的相互作用,尤其是以H亚基为筛选模型创制新农药奠定基础。     

Optimal dietary protein requirement for juvenile ivory shell, Babylonia areolate

A growth experiment was conducted to determine the optimal dietary protein requirement for juvenile ivory shell reared in indoor aerated aquaria. Six isoenergetic experimental diets using fish meal, casein and gelatin as protein sources were formulated to contain graded levels of protein (27, 33, 38, 43, 49 and 54% of dry diet, respectively). Triplicate groups of 40 shells (average weight 93.50 ± 1.70 mg) were stocked in 120-l tanks and fed to apparent satiation twice daily for 8 weeks. The results showed that the growth performance and feed utilization were significantly affected by dietary protein level (P < 0.05). Maximum weight gain, mean protein gain, specific growth rate and soft body to shell ratio occurred at 43% dietary protein level (P < 0.05). There were significant differences in protein, lipid, moisture and ash content in soft body; except that ash content in shell was not significantly affected by dietary protein level. Pepsin activity in soft body tissue significantly increased with dietary protein level up to 43%, and trypsin-like enzyme activity increased with dietary protein level up to 49%. However, lipase activity in soft body decreased with increasing dietary protein level. However, no significant differences (P < 0.05) in survival, calcium, phosphorus concentration in the shell and soft body were found among dietary treatments. Quadratic regression analysis of weight gain against dietary protein level indicated that the optimal dietary protein requirement for maximum growth and feed utilization of juvenile ivory shell is 45% of dry diet.     

Effect of dietary protein and lipid source on the growth, survival, condition indices, and body composition of marron, Cherax tenuimanus (Smith)

A trial was conducted in 12 purpose-built, commercial, drainable, earthen ponds to evaluate the effect of fish and plant protein and lipid source on the growth, condition indices, and body composition of marron (Cherax tenuimanus). Juvenile marron (1.3±0.28 S.E. g) at the stocking densities of three per square meter were fed for a period of 1 year with four different formulated isoenergetic practical diets (D₁, D₂, D₃, and D₄). Three of the test diets (D₁, D₂, and D₃) were isonitrogenous whereas the fourth test diet (D₄) was protein-free. Protein and lipid sources in D₁ were from Lupin (Lupinus albus) whereas protein and lipid sources in D₂ and D₃ were from fish meal. Fish oil (3.5%) was added to D₁, D₃, and D₄ whereas sunflower oil was added to D₂ and D₄ in order to make them isoenergetic. The four test diets were randomly allocated to three replicate ponds.

The lack of protein in D₄ did not significantly influence (P>0.05) the mean final weight and specific growth rate of marron. Survival was low in all ponds (13.82–34.66%) but feeding with D₄ resulted in a significantly (P<0.05) higher survival than marron fed with D₁ and D₂. Feeding a diet containing a combination of fish protein and fish oil (D₃) resulted in significantly higher (P<0.05) wet tail muscles-to-body weight ratio than was observed with other diets. Tail muscles protein level of all marron was significantly lower (P<0.05) at the end of the trial than at the beginning. EPA and DHA in hepatopancreas and tail muscles of marron were affected by the four test diets. The inclusion of plant protein in formulated diets had a negative impact on the pond environment due to significantly higher unionised ammonia levels that resulted in lower survival. Juvenile marron fed with a plant protein diet had significantly lower protein levels in their hepatopancreas compared to those fed with diets containing animal protein. Feeding marron with lupin protein source (D₁) and plant oil (D₂) for 1 year did not alter the lipid content of their hepatopancreas. Four test diets had no influence on the fat content of marron hepatopancreas; however, these test diets significantly reduced the protein content of the tail muscles of marron.

Environmental variables, particularly temperature, nitrogen metabolites, and the natural productivity of the ecosystem, greatly influenced the nutritional requirements of the juvenile marron under culture.     

Dopamine modulates the physiological response of the tiger shrimp Penaeus monodon

Levels of glucose, lactate, pO₂, pCO₂, HCO₃⁻, TCO₂, Na⁺, K⁺, Cl⁻, protein, and oxyhemocyanin in the hemolymph and its osmolality and pH were measured when tiger shrimp, Penaeus monodon (13.5 ± 1.5 g body weight), were individually injected with saline or dopamine at 10^− 8, 10^− 7, or 10^− 6 mol shrimp^− 1. Results showed that hemolymph glucose, lactate, pCO₂, HCO₃⁻, and TCO₂ values increased from 2 to 4 h; hemolymph osmolality, Na⁺, and total protein had increased at 2 h; and hemolymph K⁺ decreased from 2 to 8 h after the dopamine injection. All physiological parameters returned to the control values 4–16 h after receiving dopamine. The dopamine injection also significantly decreased the oxyhemocyanin/protein ratio of P. monodon which occurred at 2 h, resulting from an elevation of hemolymph protein and a slight decrease of oxyhemocyanin. These results suggest that stress-inducing dopamine caused a transient period of modulation of energy metabolism, osmoregulation, respiration, and the acid–base balance in P. monodon in adapting to this environmental stress.     

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.     

Evaluation of commercial catfish, tilapia and salmonid diets for growth promotion of white suckers

In an attempt to identify appropriate diets for use in intensive baitfish culture, the efficacy of three commercial diet formulations, a high-protein, high-lipid, low-carbohydrate diet [a semi-moist salmonid diet (SD)] and two low-protein, low-lipid, high-carbohydrate diets [a catfish diet (CD) and a tilapia diet (TD)] on growth performance of juvenile white suckers, Catostomus commersoni, was examined. All three diets permitted growth during the 16 week trial. The SD promoted highest growth and somatic protein and lipid deposition, whereas the CD and TD formulations promoted markedly higher accumulations of glycogen in the carcass and liver. The high hepatic glycogen content was associated with an elevated hepatosomatic index in the CD- and TD-fed groups. Plasma T3 levels were lower in the CD- and TD-fed groups, possibly a response to the low dietary protein or indicative of dietary insufficiency in these fish. Of the three diets evaluated, the SD was considered to be the best in terms of the growth performance parameters measured, and offers an economical and appropriate diet for intensive culture of juvenile white suckers.     

In vitro protein digestion, and growth of Atlantic salmon with different trypsin isozymes

Growth and in vitro digestion was examined in three iso-trypsin phenotypes (patterns 1, 2 and 2) of Atlantic salmon, Salmo salar. Fish grew equally well during the 128 day experiment irrespective of phenotype. At day 64 and 91, weight-specific growth rate of 2 phenotype animals were significantly lower (p < 0.05). At the end of the experiment, significant differences (p < 0.05) were noted between groups with respect to size. An in vitro method was used to examine protein digestion by the different trypsin phenotypes. Three fish meals (FM I > FM II > FM III) of high, medium and low quality, as defined by chemical analyses, were used. Phenotype 1 was able to differentiate between three fish meals ranking FM I > FM II > FM III. In contrast, phenotypes 2 and 2 were not able to separate between meals. In vitro digestion efficiency was dependent upon the three salmon types, ranking phenotype 2 > 1 = 2.     

鱼粉质量对中国对虾、真鲷、饲料转化率和蛋白质消化率的影响

IntroductionInherent to the practice of intensive aquaculture is the generation of wastes having immediate and verybroad effects on the aquatic environment.There is a growing consensus about the need to reduce waste pro-duction in aquaculture to minimize the negative impacts on the environment.Nitrogen(N) and phosphorus(P) wastes are major concerns for many aquaculture operations.In the past,formulation of diets in intensive aquaculture was often aimed at meeting requirements formaximum growt…     

Evaluation of different protein sources in fingerling grey mullet <Emphasis Type="Italic">Mugil cephalus</Emphasis> practical diets

An eight-week experiment was carried out to evaluate the effects of different protein sources (fish and haemoglobin meal, soybean meal and torula yeast), in practical diets, on growth, body composition and gut morphology of fingerling grey mullet (Mugil cephalus). Weight gain (%), SGR, FCR, N retention, PER, PGR, FDR and carcass composition of fish were not significantly affected by the dietary protein source. Fish fed the torula yeast based diet showed reduction in growth performance. Histological examinations performed on the alimentary tract of the fish showed a normal structural pattern in the experimental groups, as fundamental histological and histochemical aspects were similar if compared to the control group. The lower growth performance observed in fish fed a torula yeast based diet may be tentatively correlated with the presence of some detrimental morpho-functional aspects in the gut of these fish if compared to fish fed the other diets. Further studies are necessary to confirm this hypothesis.     

Effect of dietary protein:energy ratio on intake, growth and metabolism of juvenile green abalone Haliotis fulgens

Juvenile green abalone Haliotis rufescens were grown under laboratory conditions at 21±1 °C and fed formulated diets consisting of different protein:energy ratios (mg protein/kcal), 62, 74, 85, 100, 108, for 60 days. The level of crude protein ranged from approximately 26% to 44% while the energy content remained constant at about 4.1 kcal g⁻¹. Growth ranged from 3.63 to 12.33 mg day⁻¹. The growth of abalone fed the 100 and 108 diets was significantly greater than that of each of the other diets. Protein efficiency ratio increased as the dietary protein content increased except for the T108 diet (44% crude protein). Abalone apparently consume food to satisfy an energy requirement. Caloric expenditure due to metabolism was estimated for abalone fed diets with protein ratios of 62, 85, 100. Energy loss due to respiration did not vary appreciably among abalone fed the different diets. The proportional distribution of dietary energy into fecal, digestible, growth, and metabolic energy was estimated for abalone fed these diets. Apparent dry matter digestibility was among the lowest for abalone fed the 100 P:E diet, but growth of abalone fed this diet was significantly higher than that of each of the other treatments except the 108 diet. Unexplained energy loss to achieve balance ranged from 7% to 28.5%, some of which is probably due to differential mucus and ammonia production. Results suggest a potential for the reduction of both dietary protein and lipid without causing any adverse effects on the growth response.